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28 November 2020, Volume 36 Issue 6
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Effects of estrogen on apoptosis of spiral ganglion cells in cochlea of aged C57BL/6J mice
CHEN Long, YANG Yu-qi, FENG Zi-yi, LI Xue-rui, HAN Zi-wei, MA Ke-tao, SI Jun-qiang, LI Li
CJAP. 2020,
36
(6): 529-533. DOI: 10.12047/j.cjap.6007.2020.112
Abstract
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406
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Objective:
To observe the effects of estrogen on cochlear spiral ganglia cell apoptosis in aged C57BL/6J mice, and to explore the possible mechanism of estrogen's protective effects on senile deafness.
Methods:
Forty C57BL/6J mice were divided into the following four groups (10 mice/group): 3 m group (3 months old group), 12 m group (12 months old sham operation group); In the 12 m OVX group (ovariectomized at 12 months), bilateral oophorectomy was performed at the age of 9 months and normal feeding was performed until the age of 12 months.The 12m OVX+E2 group (estrogen intervention group) underwent bilateral oophorectomy at 9 months of age. After the one-month washout period, mice in the other groups were treated with estrogen at the dose of 100 μg/(kg·d) by subcutaneous injection, lasting 2 months to 12 months old. Mice in the other groups were fed normally.Blood samples were collected from the tail vein at the end of the treatment in 12 m OVX+E2 group. Enzyme-linked immunosorbent assays (ELISAs) was used to determine the serum estrogen levels. Auditory brainstem response (ABR) was used to detect the changes of hearing threshold in each group.Mice were anesthetized with 2% pentobarbital sodium. Bilateral cochlea was extracted after neck amputation and paraffin-embedded sections were performed.Hematoxylin eosin (HE) staining was used to observe the morphological changes in the cochlea spiral ganglion neurons (SGN), and TUNEL staining was used to observe the apoptosis of SGN. The expression levels of Caspase-3, Bax and Bcl-2 mRNA of the apoptotic proteins in cochlear spiral ganglion were measured by real-time fluorescence quantitative PCR (QRT-PCR).
Results:
Compared with the 3 m group, the hearing threshold of the 12 m group was improved, the loss of spiral ganglion cells was aggravated, and the apoptosis of the cells was increased(
P
<0.01). After removal of the ovaries, the hearing threshold of the mice in the 12 m OVX group was higher than that in the 12 m control group (
P
<0.01), and this increased threshold was accompanied by an increased loss of spiral ganglion cells, and increased apoptosis (
P
<0.01). Meanwhile, the mRNA levels of apoptotic protein Caspase-3 and Bax were increased (
P
<0.01), while the mRNA level of anti-apoptotic protein Bcl-2 was decreased (
P
<0.01). After exogenous estrogen was given to the 12 m OVX+E2 group, the hearing threshold was lower than that in 12 m OVX group(
P
<0.01). At the same time, the apoptosis of helical ganglion cells was reduced, the mRNA levels of Caspase-3 and Bax were decreased (
P
<0.01), and the Bcl-2 mRNA level was increased (
P
<0.01).
Conclusion:
Estrogen inhibited apoptosis of cochlear spiral ganglion cells in aged C57BL/6J mice ,thus achieving a protective effect on presbycusis.
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Differential expression of immediately early gene c-fos in THP-1 macrophage subtype polarization
CHEN Qi-wen, WU Guo-dong, YAN Zhi-yun, ZHANG Wen-cheng, ZHANG Mei
CJAP. 2020,
36
(6): 534-538. DOI: 10.12047/j.cjap.5865.2020.113
Abstract
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Objective:
To investigate the expression of immediate early gene c-fos in THP-1 macrophage subtype polarization.
Methods:
PMA was used to induce the polarization of THP-1 monocytes to macrophages, and the expression of c-fos in the polarization process was observed. After PMA treatment, LPS or IL-4 were used alone to induce the polarization of THP-1 macrophages to the M1 or M2 subtypes. Subsequently, real-time quantitative PCR and western-blot were used to analyze the changes in the expressions of the cell subtype markers CD274, CD86 and CD163. Meanwhile, the expression of c-fos in the polarization process was observed dynamically.
Results:
The levels of c-fos protein and mRNA expressions were up-regulated during PMA-induced polarization of THP-1 monocytes. The protein and mRNA expressions of c-fos were significantly decreased during the polarization of THP-1 cells into M1 macrophages induced by LPS. The specific markers showed the characteristics of M1 macrophages polarization at 24 h (CD86 protein increased, CD274 and CD163 protein decreased). The protein and mRNA expressions of c-fos were significantly increased during the polarization of THP-1 cells into M2 macrophages induced by IL-4. The specific markers showed the characteristics of M2 macrophages polarization at 24 h (CD86 protein decreased, CD274 and CD163 protein increased).
Conclusion:
C-fos plays an important role in the polarization of THP-1 monocytes to macrophages. Moreover, it may be involved in the regulation of macrophage subtype polarization, by inhibiting the formation of M1 macrophage and promoting the polarization of macrophages to the M2 subtype.
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Sacral nerve stimulating on intestinal mucosal immune barrier function of rats with acute complete spinal cord injury
BAI Chun-hong, ZHANG Wen-li, HU Shi-zhong
CJAP. 2020,
36
(6): 539-543. DOI: 10.12047/j.cjap.6023.2020.114
Abstract
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Objective:
To study the effects of sacral nerve root stimulation on intestinal mucosal immune barrier function in rat with acute complete spinal cord injury(SCI).
Methods:
Fifty-six Wistar rats were divided into Sham group(SG
n
=8), control group(CG 24 、 48、 72 h,
n
=8), and experimental group(EG 24、 48、 72 h,
n
=8). In CG and EG, according to Fehlings'method,we transected the spinal cord by the aneurysm clip and implanted electrodes into the third sacral foramina on the right side.We stimulated in intensity 4 V,the frequency of 15 Hz,and the pulse of 210 μs.The stimulation period was 2 hours,with 10 minutes stimulation and 10 minutes rest intermittently,twice a day at 8:00-10:00 am and 6:00-8:00 pm. The intestinal morphology was observed under light microscope and electron microscope. The protein expression levels of A20,NOD2,and CD68 by Western blot .
Results:
① SCI caused impaired intestinal epithelial barrier function. The intestinal mucosa appeared different degree of damage in CG group; cell-cell connections between intestinal epithelial cells were destroyed; The escherichia coli and other antigen translocated through the injured epithelial cell , M cells, and the leakage to the lamina propria of intestinal villi, which were improved in EG after stimulation.② The expression of A20 in EG was increased ,which had statistical differences between CG or SG(
P
<0.01); the expression ofA20 in CG was decreased, which had statistical differences between SG(
P
<0.01).The expression of NOD2 in CG was increased, which had statistical differences between SG(24 h,72 h
P
<0.05; 48 h
P
<0.01);The expression of NOD2 in EG (48 h ,72 h)was decreased, which had statistical differences between CG(48 h
P
<0.01,72 h
P
<0.05). The expression of NOD2 in EG had no statistical differences between SG. The expression of CD68 in CG was increased,which had statistical differences between SG or EG(
P
<0.01).The expression of CD68 in EG was increased in 24 h and 48 h groups ,which had statistical differences between SG(
P
<0.01),but had no statistical differences in 72 h group.
Conclusion:
Sacral nerve root 3 electrostimulation can rehabilitate the peristalsis of intestine,decrease bacterial amount,reduce inflammatory response, enhance endogenous protection, protect the intestinal mucosal immune barrier function.
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The relationship between heart rate inflection point and lactate recovery and lung function in healthy adults at the Plateau
KONG Hai-jun, ZHOU Xia, LI Xin-long, WANG Zhen-jie
CJAP. 2020,
36
(6): 544-551. DOI: 10.12047/j.cjap.6001.2020.115
Abstract
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Objective:
To explore the relationship between heart rate deflection point (HRDP) and blood lactate recovery ability and lung function of healthy people aged 20~40 years old in a plateau environment of 3 200 m.
Methods:
225 healthy people aged 20~40 who lived on a plateau of 3 200 m were used as the research subjects. The HRDP strength, heart rate recovery ability and blood lactate recovery ability were evaluated by the changes of heart rate and blood lactate recovery before, during and after the modified Conconi test.
Results:
①The heart rate of the subjects increased with the increase in exercise intensity, and the heart rate of the recovery period after exercise showed a downward trend, the rate of Conconi test center was significantly lower in male than that in female(
P
< 0.05). At the same age, male HRDP appeared later, and female HRDP appeared earlier. At the same sex stage, the time of HRDP appeared earlier in the male group with increasing age, while the phenomenon in the female group was not significant. HRDP speed had a downward trend with age. ②The inflection point concentration of blood lactic acid in the subjects gradually decreased with age, but there was no significant difference between the low-age group and the high-age group; the blood lactic acid level in the Conconi test of healthy adult males living on the plateau was significantly lower than that of females(
P
<0.05). ③FVC, MVV, FEV1 and FEV1/FVC levels in each gender group showed a downward trend with age, and the data of the male group and above were significantly higher than the female group of the same age (
P
<0.05). ④Load-Heart Rate curve and Heart Rate-Blood Lactate fitting curve showed that the correlation coefficients of the male group were 0.8345, 0.8954, 0.8680, and 0.8892 in sequence; the correlation coefficients of the female group were in sequence 0.9318, 0.9661, 0.9663 and 0.9599. HRDP values of all genders and age groups were significantly correlated with their MVV levels (
P
<0.05). Except for subjects 36~40 years old in the male group, lung function and lactate elimination rate of all genders and age groups were also significantly correlated (
P
<0.05).
Conclusion:
There are age and gender differences in exercise heart rate response rules and respiratory system functions of healthy people aged 20~40 years old living on 3 200 m plateau. There is a significant correlation between HRDP and lactic acid recovery capacity and lung function. The above indicators can be used to assess the aerobic exercise endurance ability of healthy adults living on plateau.
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Effect of walking mode on three-dimensional gait and EMG signals of human lower limbs
LIANG An-di, QIANG Jia-hao, MA Qiang, NIU Xiao-dan, ZHANG Yue, CHI Ai-ping
CJAP. 2020,
36
(6): 551-555. DOI: 10.12047/j.cjap.6021.2020.116
Abstract
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Effects of acute high altitude hypoxia on EEG power in different emotional states
CHEN Zhen, ZHANG Guang-bo, ZHOU Di, CHENG Xiang, ZHU Ling-ling, FAN Ming, WANG Du-ming, ZHAO Yong-qi
CJAP. 2020,
36
(6): 556-561. DOI: 10.12047/j.cjap.5978.2020.117
Abstract
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Objective:
To investigate the effects of acute high altitude hypoxia on EEG power in different emotional states.
Methods:
This study was two-factor within-subject design (2 levels of oxygen environment ×4 levels of emotion type). Twelve male subjects aged between 20 and 25 years old were induced to produce four different types of emotions by emotional picture evoked paradigm: low valence and low arousal(LVLA), high valence and low arousal(HVLA), low valence and high arousal(LVHA), high valence and high arousal(HVHA). Brain Products 32 was used to collect EEG signals under different emotional states. The next day, a constant depressed oxygen chamber was used to simulate a 4 300 m plateau hypoxia environment, and the same group of subjects used the same experimental paradigm to collect EEG signals 10h after hypoxia. The collected EEG signals were analyzed by power spectrum (FFT), and the five frequency bands (Delta, Theta, Alpha, beta, gamma) of the frontal lobe (F3\Fz\F4) were analyzed by variance analysis of two-factor repeated measurements.
Results:
FFT analysis found that before and after acute hypoxia, the whole brain distribution of alpha wave in four emotional states was mainly concentrated in frontal and parietal leaves; the distribution of alpha wave in the whole brain was the least in relaxed emotional state. The results of the two-factor repeated measurement ANOVA showed that: ①the power of delta\ beta band was significantly affected by the oxygen environment(
P
<0.05), and the power was enhanced under hypoxia. ②The power index of theta\ alpha band showed a significant interaction between the oxygen environment and emotional types(
P
<0.05). Except for the HVLA emotional state, the power of theta alpha band was significantly enhanced under hypoxia. ③ The two factors had no significant influence on the gamma band(
P
>0.05).
Conclusion:
Under the four kinds of emotional states, the difference of the influence of oxygen environment on brain activity was mainly in the frontal lobe, parietal lobe and part of temporal lobe. Of the four types of emotions, the oxygen environment had the least significant effect on brain activity in HVLA emotional states, while the rest showed significant differences.
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Effect of berberine on insulin resistance in diabetic rats
FAN Chun-fang, GUO Yi-sha, WANG Jia-huan, LI Ping, ZHANG Hai-long
CJAP. 2020,
36
(6): 561-564. DOI: 10.12047/j.cjap.6176.2020.118
Abstract
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253
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Effect of dapagliflozin on gene expression of glucose transporter 2 and glucose transporter 4 in kidney of type 2 diabetic rats
XU Yun, CHEN Xue-hui, BAI Li-wei, YIN Qing-feng, FENG Chun-yu, ZHANG Qing-gui
CJAP. 2020,
36
(6): 565-570. DOI: 10.12047/j.cjap.6001.2020.119
Abstract
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Objective:
To investigate the effects of dapagliflozin on the gene expressions of glucose transporter 2 (GLUT2) and glucose transporter 4 (GLUT4) in type 2 diabetic rats.
Methods:
High fat diet and 40 mg/kg streptozotocin (STZ) were used to establish the rat model of type 2 diabetes mellitus. When the fasting blood glucose (FBG) content was more than or equal to 16.7 mmol/L, the model was established successfully. After successful modeling, the rats were randomly divided into model group (group B, normal saline), dapagliflozin low-dose group (Group C, 0.75 mg/kg), dapagliflozin middle dose group (Group D, 1.5 mg/kg) and dapagliflozin high-dose group (Group E, 3.0 mg/kg), with 6 rats in each group. Six healthy SD rats were selected as normal control group (group A, normal saline). Each group was administrated by gavage once a day for 7 weeks. The body weight, serum FBG, hemoglobin A1c (HbA1c), blood urea nitrogen (BUN) and serum creatinine (Scr) were measured after 7 weeks. The levels of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) in serum and kidney were measured by enzyme-linked immunosorbent assay (ELISA). HE staining was used to observe the pathological changes of kidney. The protein expressions of GLUT2 and GLUT4 were detected by Western blot. RT-qPCR was used to detect the relative expressions of GLUT2 and GLUT4 mRNA in kidney tissue.
Results:
Compared with group A, the body weight, SOD, GSH-Px levels of rats in each group were significantly decreased (
P
<0.05), while the levels of FBG, HbA1c, BUN, SCR and MDA were significantly increased (
P
<0.05), renal pathological damage was serious, the relative expressions of GLUT2, GLUT4 mRNA and protein in renal tissue were significantly decreased (
P
<0.05). Compared with group B, the body weight, SOD, GSH-Px levels and the mRNA relative expressions of GLUT2 and GLUT4 in group C, group D and group E were significantly increased (
P
<0.05), while the levels of FBG, HbA1c, BUN, SCR and MDA were significantly decreased (
P
<0.05). The renal pathological damage in group D and group E was significantly alleviated, and the expressions of GLUT2 and GLUT4 protein in renal tissue were significantly increased (all
P
<0.05).
Conclusion:
Dapagliflozin can alleviate the condition of type 2 diabetic rats and up regulate the expression of GLUT2 and GLUT4 genes in kidney.
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Protective effect of Ginseng Guipi Pill on liver injury in rats with spleen failing to control blood syndrome and its mechanism
WANG Dan, GAO Feng, CHEN Hui, XIANG Wei-ling, LUO Zhi-xian, JIN Li-qin
CJAP. 2020,
36
(6): 571-575. DOI: 10.12047/j.cjap.5975.2020.120
Abstract
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Objective:
To investigate the therapeutic effect of Ginseng Guipi pill on rats with spleen failing to control blood syndrome and its effect on liver.
Methods:
Forty SPF male Sprague-Dawley rats were randomly divided into normal control group (
n
=10) and experimental group (
n
=30). For the first 42 days, the experimental group swam for 30 minutes every day, eating for one day and fasting for two days (diet disorder). On the 43rd to 72nd day, the rats were injected with low-molecular-weight heparin calcium to induce hemorrhage on the basis of exhaustion of swimming and diet disorder to construct a rat model of spleen failing to control blood syndrome. Those thirty rat that successfully established the model were randomly divided into 3 groups (
n
=10), model control group (MD), natural rehabilitation group (NR) and Guipi Pill group (GP). On the 73rd to 103rd day, the MD group continued to apply factors (exhausted swimming, eating disorder, and injection of low-molecular-weight heparin calcium), the NR group and the GP group stopped applying the factors , the GP group was daily administered with Ginseng Guipi Pill solution (0.2 g/ml, 10 ml/(kg·d)) , and the NR group was given an equal dose of saline. After the 103 days, blood and liver samples were collected, and the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil) and total protein (TP) content were detected, the number of red blood cells (RBC), hemoglobin (HGB) content and hematocrit (HCT) were detected, the expressions of IL-6 and TNF-α in serum were determined by ELISA kit, the levels of apoptosis-related proteins Bcl-2, Bax and Caspase3 in liver tissue were detected by Western blot. The expression levels of apoptosis-related proteins Bcl-2, Bax and Caspase3 mRNA in liver tissue were detected by real-time PCR.
Results:
Compared with the control group, the serum levels of ALT, AST and TBil in the model group were increased significantly, while the serum level of TP was reduced significantly (
P
<0.01), the number of RBC, HGB content and HCT were decreased significantly (
P
<0.01), the expressions of IL-6 and TNF-α in serum were increased (
P
<0.05), the Bcl-2 protein and mRNA levels were reduced, Bax and Caspase3 protein and mRNA levels were increased significantly (
P
<0.01); Compared with the model group, the serum levels of ALT, AST and TBil were significantly lower in the natural rehabilitation group and the Guipi Pill group, the level of TP was increased significantly (
P
<0.01), the blood RBC, HGB and HCT were increased significantly (
P
< 0.01), the expressions of IL-6 and TNF-α were decreased (
P
<0.05), the levels of Bcl-2 protein and mRNA were increased significantly, the Bax, Caspase3 protein and mRNA levels were decreased (
P
<0.05); Compared with the natural rehabilitation group, the serum levels of ALT, AST and TBil were reduced , and the TP content was increased significantly in the Guipi Pill group (
P
< 0.01), the number of RBC was increased significantly in the Guipi Pill group (
P
<0.05), the levels of Bcl-2 protein and mRNA were increased (
P
<0.05),the level of Caspase3 protein was decreased (
P
<0.05), the expression of Bax mRNA was reduced significantly (
P
<0.05).
Conclusion:
Ginseng Guipi Pill has protective effect on liver injury in rats with spleen failing to control blood syndrome. The mechanism may be related to the inhibition the liver cell apoptosis and the release of pro-inflammatory cytokines.
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Roumudan formulation attenuated liver fibrosis by inhibiting TGF-β1/Smad4 pathway in mice
LI Qian, YAN Shu-guang, HUI Yi, LI Jing-tao, WEI Hai-liang, ZHANG Hong
CJAP. 2020,
36
(6): 576-581. DOI: 10.12047/j.cjap.6010.2020.121
Abstract
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Objective:
To investigate underlying mechanism involving Roumudan(RMD) formulation (Z20160012) suppressed liver fibrosis induced by CCl
4
injection in mice by inhibiting TGF-β1/Smad4 pathway.
Methods:
Male BALB/c mice were randomly divided into control group, liver fibrosis model group and RMD-treated group(
n
=11). Mice in liver fibrosis model and RMD-treaded groups were injected intraperitoneally with CCl
4
(20% in olive oil) at the dose of 2.5 mL/kg two times for one week and 5 mL/kg two times for 4 weeks. Mice in control group were treated intraperitoneally with the same volume of olive oil at the same time intervals. From sixth week, Mice in liver fibrosis model group were administrated with CCl
4
(20% in olive oil, 1.5 ml/kg once per week) intraperitoneally and given distilled water by intragastric gavage. Mice in the RMD-treated group were administrated with CCl
4
(20% in olive oil, 1.5 ml/kg/mouse once per week) intraperitoneally and given RMD(6.2 g/kg everyday) by intragastric gavage. Mice in the control group were administrated with olive oil (1.5 ml/kg/mouse once per week) intraperitoneally and given distilled water by intragastric gavage. The serum AST and ALT levels were estimated for assessment of liver function. The pathologic changes of mice' livers were examined by the HE, Masson, immunohistochemical staining, Western Blot, Q-PCR and so on.
Results:
After intraperitoneally injected with CCl
4
in mice, the pathological characteristics of liver fibrosis were observed compared with the control group at the sixth week. Compared with the liver fibrosis model group, RMD improved the liver function significantly through reducing liver index(
P
<0.01) and the levels of ALT(
P
<0.01), AST(
P
<0.01) and HYP(
P
<0.05). The expression of TGF-β1(
P
<0.05), α-SMA(
P
<0.05),
COL1A1
(
P
<0.01) and
COL3A1
(
P
<0.01) were decreased by RMD. The positive expression area of
Smad4
mRNA in RMD treated group was lower than that in liver fibrosis model group.
Conclusion:
The RMD formulation could attenuate liver fibrosis by inhibiting TGF-β1/Smad4 pathway and extracellular matrix (ECM) production in mice.
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Effects of
Biejia Yugan Granule
on rat with ethanol-induced hepatic fibrosis
WEI Ling-xia, DING Mao-peng, WANG Zhi-wang, LIU Xue-feng, PANG Ya-rong, GUO Mei
CJAP. 2020,
36
(6): 582-586. DOI: 10.12047/j.cjap.6109.2020.122
Abstract
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Objective:
To investigate the prevention and treatment effect of
Biejia Yugan Granule
(BYG) on ethanol-induced hepatic fibrosis (EHF) rats.
Methods:
SD rats were randomly divided into blank control group, model group, BYG group, colchicine group and BYG low and BYG high dose groups (
n
=8). The EHF rat model was established by intragastric edible ethanol with a gradually increased dose. Briefly, the rats of model group, colchicine group and BYG low and high dose groups were given gavage of 5 g/(kg·d) ethanol at week 1~4, 7 g/(kg·d) ethanol at week 5~8, 9 g/(kg·d) ethanol at week 9~12 and 9.5 g/(kg·d) ethanol at week 13~24. And the other two groups were treated with equal volume water. At the same time, the corresponding drugs were administrated daily: BYG group was treated with
Biejia Yugan Granules
5.55 g/kg, colchicine group was treated with colchicine 0.1 mg/kg, BYG low-dose and high-group were treated with
Biejia Yugan Granules
1.85 and 5.55 g/kg respectively. The blank control group and model control group were given the same amount of purified water. On the 169th day of the experiment, the effects of BYG on the macroscopic changes of rat liver organs, the water content of liver tissue and the pathological changes of fibrosis, the content of hydroxy proline (Hyp) in liver tissue and the expression levels of α-SMA and CREB were observed.
Results:
BYG at the doses of 1.85 and 5.55 g/kg could significantly improve the macroscopic changes of liver and pathological changes of liver tissue fibrosis in rats with EHF, reduce the contents of water and Hyp in liver tissue, and down-regulate the expressions of α-SMA and CREB.
Conclusion:
BYG has obvious effect on inhibiting EHF and one of its mechanisms is down-regulate the content of CREB.
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Changes in blood flow during cerebral ischemia/reperfusion and their correlation with cerebral edema in rats
ZHANG Ran, MY Meng-yao, SU Xin-yu, MENG Xiang, JIANG Kun-peng, LI Shu, HONG Yun
CJAP. 2020,
36
(6): 586-589. DOI: 10.12047/j.cjap.6049.2020.123
Abstract
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324
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Effect of 36h total sleep deprivation on the object working memory: an ERP study
PENG Zi-yi, CHEN Xue-wei, ZHANG Ying, YANG Ye-bing, ZHANG Li-wei, SHAO Yong-cong
CJAP. 2020,
36
(6): 590-594. DOI: 10.12047/j.cjap.6030.2020.124
Abstract
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Objective:
To investigate the effects of 36 h total sleep deprivation (TSD) on object working memory by event related potential(ERP).
Methods:
We used a pre-post-design, sixteen healthy college students (age range: 21-28 years, mean age: 23 years) received object working memory tasks while awake and after 36 hours of TSD and simultaneously recording electroencephalograph (EEG) data while completing 2-back object working memory tasks. ERP data were statistically analyzed using repeated measurements analysis of variance to observe the changes in the working memory-related P2, N2 and P3 components.
Results:
After 36 h TSD, the latency of N2 waves related to object working memory significantly was prolonged (
P
<0.05), and the amplitude was decreased, but difference did not reach statistical significance (
P
>0.05). The latency of P2 was significantly prolonged after TSD (
P
<0.05). There was no significant difference in the change of latency and amplitude of P3 waves (
P
>0.05).
Conclusion:
36 h of total sleep deprivation affected working memory-related components and impaired object working memory capacity.
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Effects of low intensity resistance training of blood flow restriction with different occlusion pressure on lower limb muscle and cardiopulmonary function of college students
LU Jie-ming, LIU Shan-yun, SUN Peng, LI Wu-lan, LIAN Zhi-qiang
CJAP. 2020,
36
(6): 595-599. DOI: 10.12047/j.cjap.6032.2020.125
Abstract
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Objective:
The study compared the effects of low intensity resistance training with blood flow restriction (BFR) with different occlusion pressure on lower limb muscle and cardiopulmonary function.
Methods:
Twenty-seven college students were randomly divided into three groups by different occlusion pressure: 0 mmHg (group C), 120 mmHg (group L) and 180 mmHg (group H). Before and after training (3 times a week for 12 weeks) with an inflatable cuff (20% 1RM, half squat), the muscle thickness(MTH)of rectus femoris and medius femoris, relative peak knee extensor moment(rM), peak power(P), relative maximal oxygen uptake(rVO
2
max), stroke volume(SV), cardiac output(CO), ejection fraction(EF) and other indicators were measured for all subjects.
Results:
When compared with pre-training, and rectus femoris, the MTH of medius femoris, rM, rVO
2
max, SV, CO and EF were significantly increased in group L and group H after 12 weeks training(
P
<0.05,
P
<0.01), as well as compared with group C after training(
P
<0.05,
P
<0.01). There was no significant difference between group L and group H after training.
Conclusion:
BFR training protocols under 120 mmHg or 180 mmHg pressure were effective in improving muscle and cardiopulmonary function.
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Effects of miR-203a targeting and its target gene on lymph node metastasis of breast cancer
HAN Li-rong, SONG Jiang-qin, GUO Fei-bo, ZHANG Dong-wu
CJAP. 2020,
36
(6): 600-604. DOI: 10.12047/j.cjap.6041.2020.126
Abstract
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Objective:
To study the expression and correlation of mir-203a and its target gene ATM in breast cancer tissues, so as to provide theoretical basis for the pathogenesis of breast cancer, especially lymph node metastasis.
Methods:
Thirty paired breast cancer and paracancer normal tissues were collected, and RT-qPCR was used to detect the relative expression levels of mir-203a and ATM in the samples of the two groups. Correlation analysis was conducted for mir-203a and ATM, and correlation analysis was conducted for the pathological characteristics, so as to compare whether there were statistical differences between mir-203a and ATM in lymph node metastasis and non-metastasis.
Results:
Compared with normal paracancer tissues, the expression level of mir-203a in breast cancer tissues was significantly increased (
P
<0.01), and the expression level of ATM was significantly decreased (
P
<0.01), showing a significant negative correlation between the two tissues (
r
=-0.847,
P
<0.01).The expression level of mir-203a and ATM was significantly correlated with lymph node metastasis and different clinical stages (
P
<0.05). The expression level of mir-203a in the group with lymph node metastasis were significantly lower than that in the group without lymph node metastasis (
P
<0.05), and the expression of ATM in the group with lymph node metastasis was significantly higher than that in the group without lymph node metastasis (
P
<0.01).
Conclusion:
The overexpression of mir-203a in the early stage of breast cancer may inhibit the expression of its target gene ATM, which may be a protective mechanism to regulate the proliferation,metastasis and invasiveness of tumor cells. In the middle and late stage, mir-203a is down-regulated and the ATM gene is up-regulated, which may be involved in lymph node metastasis.
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Effects of Bushen Zhuanggu Granules on expressions of serum GH with IGF-1 and its receptors in bone tissue of ovariectomized rats
SU Hai-rong, CHENG Wen-yao, YUAN Qiu-hong, OUYONG Jing, DENG Wei-min
CJAP. 2020,
36
(6): 605-610. DOI: 10.12047/j.cjap.6067.2020.127
Abstract
PDF
(2137KB) (
238
)
Objective:
To investigate the effects of Bushen Zhuanggu granule on the expressions of serum growth hormone (GH) and insulin-like growth factor-1(IGF-1) and their receptors in bone tissues of ovariectomized rats.
Methods:
Forty-eight SD female rats (weight 273±21.3 g) were divided into 4 four groups: the dosage of Bushen Zhuanggu granule group (BSZG) was 2.5 g/(kg·d),the do -sage of estradiol group(E
2
) was 0.071mg/(kg·d),sham group (SHAM) and ovariectomized model group (OVX group) were given the same amount of saline by oral administration.Each group included 12 rats,the treatments were conducted once a day. After 3 and 6 months of treatment,bone mineral density (B -MD) was measured by bone density instrument;the serum levels of GH and IGF-1 were detected by EL-ISA;the expressions of GHR and IGF-1R of bone tissue were detected by qPCR;the optical density(OD)value and positive cell count of pituitary GH immunohistochemical tablets were analyzed by Image J software,respectively.
Results:
①After 3 months intervention,compared with the SHAM, the BMD of the spine in E
2
group was increased(
P
<0.05),and the BMD of both parts in BSZG group were increased(
P
<0.05).After two stage intervention,BMD of both part in the two drug groups was higher than that in the OVX group(
P
<0.05).②After two-stage intervention,the expression levels of serum GH and IGF-1,GHR and IGF-1R in BSZG group were higher than those in OVX group (
P
<0.05).The levels of serum GH and it's receptors in the E
2
group were increased (
P
<0.05), but the serum IGF-1 level remained unchanged (
P
>0.05) or even was decreased (
P
<0.05).③After two stage in -tervention,the OD values and the positive cells count in the two drug intervention groups were increased (
P
>0.05).④Pearson correlation analysis showed that serum GH,IGF-1 concentration and it's receptors in bone tissue were positively related with BMD. Serum GH concentration was positively correlated with OD values and number of positive cells.
Conclusion:
Bushen Zhuanggu granule can be used to improve the expressions of GH,IGF-1 in serum and its receptors in bone tissues of ovariectomized osteoporosis rats to prevent further loss of bone mass and increase bone mineral density.
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The role of syndecan-1 in the transformation of pulmonary epithelial stroma in rats with chronic obstructive pulmonary disease and its mechanism
JIN Yan, LIU Wen-ming
CJAP. 2020,
36
(6): 611-615. DOI: 10.12047/j.cjap.6001.2020.128
Abstract
PDF
(1625KB) (
214
)
Objective:
To explore the role and mechanism of polysaccharide-1 (syndecan-1) in the transformation of lung epithelial stroma (EMT) in rats with chronic obstructive pulmonary disease (COPD).
Methods:
Thirty male SD rats of clean grade were randomly divided into sham operation group (normal saline injection after tracheal exposure,
n
=10), COPD group (fumigation after injection of 1 mg/ml lipopolysaccharide, transfection of 100 μl empty virus,
n
=10) and syndecan-1 overexpression group (fumigation after injection of 1 mg/ml lipopolysaccharide, and transfection of 100 μl carrying rat syndecan-1 gene Ad-CMV-GFP-SDC1,
n
=10), once a day for two weeks. After the treatment, the lung function was detected and lung tissues were collected. HE staining was used to observe lung injury. The expression levels of syndecan-1, vimentin and E-cadherin in lung tissue of rats in each group were detected by immunohistochemistry. Western blot was used to detect the expressions of TGF-β1, Smad2/3 and p-Smad2/3. The mRNA levels of vimentin, E-cadherin, TGF-β1 and Smad2/3 were detected by
q
RT-PCR.
Results:
Compared with the sham operation group, the airway mucosa of COPD group was exfoliated, the lumen was narrow, the airway wall was more inflammatory cell infiltration, emphysema was serious, expiratory volume (V
E
), peak expiratory flow (PEF), Forced expiratory volume in 0.3 s (FEV
0.3
) and the expression of E-cadherin mRNA were significantly decreased (
P
<0.05). While the expression of vimentin, TGF-β1 and Smad2/3 mRNA were increased, and the expression levels of TGF-β1, Smad2/3 and p-Smad2/3 protein were significantly increased (
P
<0.05). Compared with COPD group, the number of airway mucosa exfoliation and inflammatory cell infiltration in airway wall were decreased, lumen stenosis and emphysema were improved, the levels of V
E
, PEF, FEV
0.3
and the expression of E-cadherin mRNA were significantly increased (
P
<0.05). And the expressions of vimentin, TGF-β1 and Smad2/3 mRNA were decreased, and expression levels of TGF-β1, Smad2/3 and p-Smad2/3 protein were significantly decreased (
P
<0.05).
Conclusion:
In COPD rats, TGF-β/Smad signal pathway activation induced the production of EMT; overexpression of syndecan-1 could inhibit the EMT mediated by TGF-β/Smad signal pathway, and improve the lung tissue injury of COPD rats.
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Toxic effects of Vitamin C combined with temozolomide on glioma cells and its mechanism
CHEN Ming-sheng, ZHAO Hai-kang, CHENG Ying-ying, YUAN Zhi-hai, ZHANG Yue-lin
CJAP. 2020,
36
(6): 616-621. DOI: 10.12047/j.cjap.5958.2020.129
Abstract
PDF
(1326KB) (
212
)
Objective:
To investigate the toxic effects of vitamin C (VC) combined with temozolomide (TMZ) on gliomas and its mechanism.
Methods:
Human glioma cells BMG-1 and SHG44 cells were cultured
in vitro,
specifically divided into control group (without VC and TMZ), TMZ group (0.2 mmol/L), VC (0.5 mmol/L)+TMZ(0.2 mmol/L) group and TMZ(0.2 mmol/L TMZ)+U0126(10 μmol/L)group, each experiment was repeated three times. Cell survival rate was detected by MTT assay; Cell apoptosis was detected by flow cytometry and Annexin V-FITC/PI staining; Reactive oxygen species (ROS) levels were detected by ROS detection kit, and Western blot was used to detect the expression levels of proteins related to apoptosis, autophagy and ERK pathway.
Results:
Compared with the control group, the survival rate of glioma cells in the TMZ group was decreased significantly(
P
<0.05). Compared with the TMZ group, the survival rate of glioma cells in the VC+TMZ group was decreased significantly(
P
<0.01), the cell apoptosis rate was increased, and the expressions of Bax, Cleaved caspase-3 and Cleaved PARP protein were increased, while the expression of Bcl-2 was decreased. The ROS level and autophagy rate were decreased, while the expression of LC3-II/LC3-1 was decreased, and the expression of p62 was increased in the VC+TMZ group (all
P
<0.05). At the same time, VC combined with TMZ decreased the expression level of p-ERK1/2-related protein in BMG-1 and SHG44 cells, and increased the apoptosis rate (
P
<0.05).
Conclusion:
VC combined with temozolomide can enhance the toxicity of glioma cells. This effect is to promote apoptosis and inhibit temozolomide-mediated autophagy through the regulation of the ERK signaling pathway.
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Effects of LncRNA UNC5B-AS1 on adhesion, invasion and migration of lung cancer cells and its mechanism
TAN Jian-Jun, LONG Shu-Zi, ZHANG Tao
CJAP. 2020,
36
(6): 622-627. DOI: 10.12047/j.cjap.5993.2020.130
Abstract
PDF
(1350KB) (
236
)
Objective:
To investigate the effects of long-chain non-coding RNA (lncRNA) UNC5B-AS1 on the adhesion, invasion and migration of lung cancer cells by regulating the expression of miR-218-5p.
Methods:
Twenty specimens of lung cancer patients and corresponding paracancerous tissues were surgically removed and collected from the oncology department of Chongqing Three Gorges Central Hospital from June 2017 to June 2019. Real-time quantitative PCR (qRT-PCR) was used to detect the expressions of UNC5B-AS1 in human bronchial epithelial cells HBE and different lung cancer cells of A549, H1437, H1975, H1299 and H460. UNC5B-AS1 siRNA was transfected into lung cancer A549 cells. Adhesion assay, transwell invasion assay and scratch assay were used to detect the effect of UNC5B-AS1 on adhesion, invasion and migration of A549 cells. qRT-PCR and dual luciferase reporter gene were used for the detection and identification of UNC5B-AS1 targeting miR-218-5p. The expression of epithelial-mesenchymal transition (EMT)-related protein was detected by Western blot.
Results:
The expression of UNC5B-AS1 in lung cancer tissues and cells was significantly higher than that in adjacent tissues and bronchial epithelial cells (
P
<0.05). The expression of UNC5B-AS1 in lung cancer A549 cells was the highest (
P
<0.05). Down-regulation of UNC5B-AS1 expression inhibited adhesion, invasion and migration of A549 cells (
P
<0.05). qRT-PCR and dual luciferase reporter assay experiments showed that UNC5B-AS1 targeted the regulation of miR-218-5p expression. Down-regulation of UNC5B-AS1 inhibited E-cadherin protein expression and promoted Vimentin and Twist protein expression.
Conclusion:
lncRNA UNC5B-AS1 promotes adhesion, invasion and migration of lung cancer cells through targeted regulation of miR-218-5p expression, and its mechanism may be related to the promotion of EMT.
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Effect of betulinic acid on proliferation of human gastric cancer SGC-7901 cells
CHEN Ya-nan, SHAO Shu-li, HE Meng-qi, HUANG Xin, ZHANG Wei-wei, ZHANG Zhen-zhu
CJAP. 2020,
36
(6): 628-632. DOI: 10.12047/j.cjap.6036.2020.131
Abstract
PDF
(1142KB) (
129
)
Objective:
Human gastric cancer SGC-7901 cells were treated with betulinic acid(BA)at the concentrations of 0, 10, 20, and 30 μg/ml, and treated with conventional chemotherapeutic drug 5-Fu as a positive control to explore its effect on cell proliferation. Trypan blue and GIEMSA staining method were used to investigate the effect of BA on cell growth inhibition and clone formation. EdU method and flow cytometry were used to explore the proliferation and cell cycle of SGC-7901 cells after treated with BA, respectively. qRT-PCR and Western blot were also applied to determine the mRNA and protein levels of cyclin D1 and cyclin B1.
Results:
The cell growth inhibition rate was increased after treated with different concentrations of BA in SGC-7901 cells(
P
<0.05). After treated for 48 h, BA decreased the clone information and cell proliferation of SGC-7901 cells markedly in dose-and time-dependent manners (
P
<0.01). Flow cytometry analysis showed that BA obviously increased the proportion of SGC-7901 cells in G1 phase but decreased the proportion of those in S phase. qRT-PCR and Western blot analysis showed that the mRNA and protein levels of cyclin D1 and cyclin B1 were significantly downregulated by BA at different concentrations(
P
<0.01). Compared with the 5-Fu control group, when the concentration of BA was 20 μg/ml and 30 μg/ml, the cell proliferation ability was significantly decreased, the cell cycle was inhibited, and the expression of cyclin was reduced (all
P
<0.05).
Conclusion:
The betulinic acid regulates the proliferation of SGC-7901 cells by inhibiting the expressions of cyclin D1 and cyclin B1, which leads to cell cycle arrest and proliferative inhibition.
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Effects of Toosendanin on the formation of CTPS cytoophidium in human gastric cancer cell MKN-45 and its mechanism
CHEN Wen, ZHANG Wei-wei, PAN Yang, LIU Chang, SHAO Shu-li
CJAP. 2020,
36
(6): 633-636. DOI: 10.12047/j.cjap.6026.2020.132
Abstract
PDF
(1022KB) (
140
)
Objective:
To investigate the relationship between toosendanin(TSN) and CTP synthase(CTPS) cytoophidium formation in gastric cancer MKN-45 cells.
Methods:
In this study, the experimental material is MKN-45 human gastric cancer cells. It contains 7 treatment groups of 0, 20, 40, 60, 80, 100, and 120 nmol/L TSN. Each group was treated in triplex privately for 24、48 and 72 hours. Cell counting kit-8 (CCK8) was used to detect the inhibitory effect of TSN on the proliferation of MKN-45 cells. After immunofluorescence detection, the morphology of CTPS cells was observed by a laser confocal microscope. The effect of TSN on MYC gene expression was detected by qRT-PCR. In addition, it contains 2 treatment groups of 1 mmol/L DON and 1 mmol/L MPA, each group was treated in triplex privately for 6 hours and then the cytoophidium morphology was detected by immunofluorescence.
Results:
The results of immunofluorescence showed that CTPS formed a filamentous cytoophidium structure after treating MKN-45 cells with 1 mmol/L DON and 1 mmol/L MPA, which means that the cells have the ability to form CTPS cytoophidium; The cell proliferation rate of TSN treatment group was significantly lower than that of 0 nmol / L TSN group (
P
<0.01); Immunofluorescence results showed that CTPS cytoophidium was the most abundant in MKN-45 cells after treated with 80 nmol/L TSN for 72 h. The results of qRT-PCR showed that MYC expression in cells was significantly decreased after treated with 80 nmol/L TSN for 24 h (
P
<0.05), and MYC expression was significantly increased after 48 h (
P
<0.01), and then decreased.
Conclusion:
Toosendanin may affect intracellular cytoophidium assembling by regulating the expression of MYC.
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miRNA-191 promotes proliferation, migration and invasion of prostate cancer by targeting PLCD1
NIE Yong-li, TANG Shi-min, PENG Fang, XU Tao, MAO Yong-rong
CJAP. 2020,
36
(6): 637-641. DOI: 10.12047/j.cjap.6031.2020.133
Abstract
PDF
(1333KB) (
258
)
Objective:
To investigate the effects of miRNA-191 on the proliferation, migration and invasion of prostate cancer, and to explore its mechanism.
Methods:
The expression levels of miRNA-191 in four human prostate cancer cell lines (PC-3, DU-145, LNCa P, 22RU1) and human normal prostate cell line RWPE-2 were detected, and prostate cancer cell line PC-3 was selected as the experimental object. PC-3 cells were divided into three groups: blank control group (no transfection), miRNA-191 NC group (PC-3 cells transfected with Inhibitor NC) and miRNA-191 Inhibitor group (PC-3 cells transfected with miRNA-191 Inhibitor), and each group was provided with three multiple pores. The expression levels of miRNA-191 and PLCD1 were detected by RT-PCR. The cell proliferation was detected by CCK8 assay. Scratch test and invasive test were used to detect cell migration and invasive ability. Through Targetscan target gene prediction website, PLCD1 was screened as the target protein of miRNA-191, and verified by double luciferase target experiment.Western blot assay was used to detect the expression of PLCD1 in cells of each group.
Results:
Compared with RWPE-2 cells, the expression level of miRNA-191 in human prostate cancer cells was significantly higher (
P
<0.05), and the expression level of miRNA191 in PC-3 was significantly higher than that in other three cell lines (
P
<0.05). After inhibiting the expression of miRNA-191, the expression levels of PLCD1 was significantly higher while PC-3 cells' proliferation ability was inhibited, and their migration and invasion ability were significantly lower than those of blank control group and miRNA-191 NC group (
P
< 0.05). The results of double luciferase reporter gene assay showed that PLCD1 gene was a target gene of miRNA-191.
Conclusion:
miRNA-191 promote the proliferation, migration and invasion of prostate cancer PC-3 cells by targeting PLCD1.
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Effects of SmacN7 inducing apoptosis of breast cancer cell MDA-MB-157 and its mechanism
LYU Zi-yan, ZHANG Xiao-yan, CHEN Hong, JIANG Pan-ruo, CHEN Jia-yu
CJAP. 2020,
36
(6): 642-647. DOI: 10.12047/j.cjap.5948.2020.134
Abstract
PDF
(1092KB) (
414
)
Objective:
To investigate the effects and molecular mechanisms of Second mitochondria-derived activator of caspase N7 (SmacN7) on the apoptosis of breast cancer cells MDA-MB-157.
Methods:
Breast cancer cells MDA-MB-157 were treated with SmacN7 at the concentrations of 0-20 μmol/L. The proliferation activity of the cells was detected by MTS method, apoptosis and cell cycle were analyzed by flow cytometry, karyotypic changes of MDA-MB-157 cells were observed by Hoechst33342 staining, mitochondrial membrane potential was detected by JC-1 staining, and LDH release experiment was used to detect the drug cytotoxicity. Real time PCR was used to analyze the transcription levels of genes in MDA-MB-157 cells. The effect of inhibiting breast cancer proliferation was confirmed by tumor inhibition experiments.
Results:
After treated with SmacN7, the inhibition rate of proliferation and apoptosis rate of breast cancer cells MDA-MB-157 were increased (
P
<0.01), the karyotype changed significantly, the mitochondrial membrane potential in cells was decreased, and the LDH release was increased. The transcription levels of TRAIL, DR4, DR5, p53, PARP-1, Bax, Bid, BAK, caspase-3, caspase-8 and caspase-9 were up-regulated (
P
<0.01), and the transcription levels of Ras, PI3K, AKT, mTOR, Bcl-2, Bcl-xL, MCL-1, Survivin, cIAP-1 and cIAP-2 were inhibited (
P
<0.01).
Conclusion:
SmacN7 induces apoptosis of breast cancer cell MDA-MB-157 through TRAIL-mediated death receptor pathway and mitochondrial-mediated endogenous apoptosis pathway, and plays a role in anti-breast cancer.
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Effect of thyroxine on the expression of HIF-1α after aneurysmal subarachnoid hemorrhage in rat brain and its mechanism
RAN Hui,YIN Hao, LIU Chuang-xi , HAN Guo-qiang, GAO Fang-you, SHEN Hong-bin , FU Hang , XU Xiao-zhong, LI Tao, MA Jun
CJAP. 2020,
36
(6): 648-652. DOI: 10.12047/j.cjap.5985.2020.135
Abstract
PDF
(1150KB) (
296
)
Objective:
To investigate the effect of thyroxine (T4) on the expression of hypoxia inducible factor-1α (HIF-1α) in rat brain after aneurysmal subarachnoid hemorrhage (SAH) and its mechanism.
Methods:
Seventy-two adult male SD rats were randomly divided into the following 4 groups: subarachnoid hemorrhage model group(SAH), subarachnoid hemorrhage model and T4 group (SAH with T4), subarachnoid hemorrhage model with normal saline group (SAH with vehicle), and sham-operation group, 18 rats in each group. The model of subarachnoid hemorrhage group was established by internal carotid artery puncture. CT plain scan was performed after the modeling immediately, T4 was administrated by intraabdominal injection of 3 μg/100 g every 24 hours for 3 days. SAH with T4 group was treated with thyroxine. SAH with vehicle group was treated with equal volume vehicle, all of them were killed 72 hours after modeling. The brain water content was determined to evaluate the brain edema, the apoptosis of cerebral cortex cells was detected by TUNEL method, and HIF-1α protein and p-Akt protein in cerebral cortex were detected by immunohistochemistry in six SD rats of each group.
Results:
After the modeling, the brain tissues of SAH group, SAH + T4 group and SAH +vehicle group were swollen obviously, and blood clots were observed in subarachnoid space. The neurobehavioral score,the brain water content, apoptosis index, HIF-1α protein and p-Akt protein in SAH group were significantly higher than those in sham-operation group(
P
<0.05).The neurobehavioral score,HIF-1α protein and p-Akt protein in SAH with T4 group were significantly higher than those in SAH group, and the brain water content, apoptosis index were significantly lower than those in SAH group (
P
<0.05).
Conclusion:
The expression of HIF-1α protein in the brain of rats after aneurysm subarachnoid hemorrhage can be upregulated by T4 replacement therapy, which may by activating the signal pathway of inositol triphosphate kinase / protein kinase B (PI3K/Akt). Finally, apoptosis index was decreased, the rat behavior was improved and the brain was protected.
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The effect of parisib on chondrocytes in osteoarthritis rats
LI Gang, JI Yin-xi, GAO Yi, FENG Yan-bing
CJAP. 2020,
36
(6): 652-655. DOI: 10.12047/j.cjap.6024.2020.136
Abstract
PDF
(1240KB) (
91
)
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Protective effect and immune mechanism of berberine on cerebral ischemia/reperfusion injury in rats
SUN Ke, LUO Zhao-liang, HU Chen, GONG Ting-liang, TANG Guo-hua, WU Shao-ping
CJAP. 2020,
36
(6): 656-661. DOI: 10.12047/j.cjap.6001.2020.137
Abstract
PDF
(1552KB) (
451
)
Objective:
To investigate the protective effect and immune mechanism of berberine on cerebral ischemia/reperfusion injury in rats.
Methods:
Fifty SD rats were randomly divided into sham operation group (Sham), model group (Model), berberine low dose groups (BBR-L, 25 mg/kg), berberine medium dose groups (BBR-M, 50 mg/kg) and berberine high dose groups (BBR-H, 100 mg/kg), with 10 rats in each group. Longa suture method was used to establish a rat model of cerebral ischemia/reperfusion, after 2 hours of ischemia, reperfusion for 24 hours. Rats in BBR-L, BBR-M and BBR-H were treated with berbrerine by gavage 2 hours after successful model building, while the sham operation group and the modle group were given the same volume of saline as described above. After 24 hours of administration, the activity of antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), cytokine tumor necrosis factor α (TNF-α) , interferon β (IFN-β) , interleukin 6 (IL-6) and nitric oxide (NO) content were detected by ELISA assay. Serum CD4
+
, CD8
+
and CD4
+
/CD8
+
contents were measured by flow cytometry to investigate the immune function of each group. RT-qPCR and Western blot were used to detect NF-kappaB-NOD-like receptors 3 (NF-κB-NLRP3) signal axis key genes and protein expression in rat brain tissue.
Results:
Compared with the sham operation group, the degree of neurological deficit and the rate of cerebral infarction were increased in the model group (
P
<0.05), and the levels of serum NO, TNF-α, IFN-β, IL-6, NF-κB p65, NLRP3, ASC and caspase-1 in brain tissue were increased (
P
<0.05), while the activities of SOD, GSH-Px and the levels of CD4
+
, CD8
+
and CD4
+
/CD8
+
in serum were decreased (
P
<0.05). Compared with the model group, the degree of neurological deficit and the rate of cerebral infarction were increased in the BBR-H, BBR-M and BBR-L groups (
P
<0.05), and the levels of serum NO, TNF-α, IFN-β, IL-6, NF-κB p65, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) and caspase-1 in brain tissue were increased (
P
<0.05), while the activities of SOD, GSH-Px and the levels of CD4
+
, CD8
+
and CD4
+
/CD8
+
in serum were decreased (
P
<0.05).
Conclusion:
Berberine may reduce oxidative stress, inhibit inflammation, enhance immune function, and reduce cerebral ischemia/reperfusion injury in rats, which may be related to the inhibition of NF-κB-NLRP3 signaling.
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The effect of nanoscale zirconium-porphyrin metal-organic framework on zebrafish embryonic neurodevelopment
WANG Ya-jie, YUAN Bo, LIU Wei, LI Yu-hao, YUAN Xiao-yong
CJAP. 2020,
36
(6): 662-667. DOI: 10.12047/j.cjap.6015.2020.138
Abstract
PDF
(1769KB) (
155
)
Objective:
To investigate the effect of nanoscale zirconium-porphyrin metal-organic framework (NPMOF) on the development of nervous system in larval zebrafish.
Methods:
Embryos of zebrafish were incubated to E3 medium (
n
=500) or 100 mg/L NPMOF-E3 medium (
n
=500) from 6 hours post fertilization (hpf) to 28, 48, 72, 96 or 120 hpf. At 28, 48, 72, 96 and 120 hpf, 60 fish were collected respectively for quantitative real-time PCR in both groups. At 120 hpf, 20 fish were used for
in situ
hybridization, 150 fish were used for immunofluorescence and 30 fish were used for behavioral test, respectively. The shape and size of NPMOF was measured by TEM, and the optical properties were detected by UV-Vis and Fluorescence Spectrometer.
In vivo
development of multiple neurocytes was examined via
in situ
hybridization, immunofluorescence and quantitative real-time PCR. Behavioral test was used to manifest the locomotor changes of larval zebrafish.
Results:
Compared to control group, the mRNA expression levels of neurodevelopment-relative, neuron-relative and neuroglia-relative genes were partially increased obviously after NPMOF exposure (
P
< 0.05). The distribution and phenotype of neurons and oligodendrocytes showed no significant differences between exposed and unexposed groups, while exposed group showed an increase in the number of müller glia and astrocytes (
P
<0.05). In behavioral test, there was an increase in total movement distance, fast movement time and velocity and a decrease in total rest time following NPMOF exposure (
P
<0.05).
Conclusion:
The data indicate the potential facilitating effect of 100 mg/L NPMOF on neurodevelopment
in vivo
, especially on the growth of müller glia and astrocytes.
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Simultaneous determination of three antidepressant drugs in feces by HLPC-MS
SHAO Yi-fan, ZHANG An-qi, CHEN Tian-jiao, LI Hai-bei, CHEN Zheng-shan, YIN Jing, YANG Dong, SHI Dan-yang, LIANG Yong-bing, CHENG Chun-yan, TAN Rong, SUN Dong-liang, LI Jun-wen, JIN Min
CJAP. 2020,
36
(6): 668-672. DOI: 10.12047/j.cjap.6173.2020.139
Abstract
PDF
(1173KB) (
143
)
Objective:
To establish a high performance liquid chromatography tandem mass spectrometry (HPLC / MS) method for the simultaneous determination of three antidepressant drugs in feces.
Methods:
Samples were pretreated with n-hexane isopropanol (95:5, v/v). Gradient elution was carried out with mixed liquid of ultrapure water and acetonitrile as mobile phase and separated by Agilent ZORBAX SB-C18 liquid chromatography column (2.1 mm×100 mm, 3.5 m). The samples were detected by electrospray ionization tandem mass spectrometry and quantified by internal standard method.
Results:
The recoveries of duloxetine, fluoxetine and escitalopram in fecal samples were 61.6% - 116.5%, with precision of 2.80% - 12.9% (
n
=5). The correlation coefficients (r) of linear equations were all greater than 0.995. The detection limits were 0.1, 1, and 0.001 μg/g, and the limits of quantification were 0.5, 2 and 0.005 μg/g, respectively.
Conclusion:
The method is simple and accurate to detect the contents of three antidepressants in feces, such as duloxetine, fluoxetine and escitalopram.
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