To examine the effects of ischemia and anoxia on cell activation and cell cycle of astrocytes in vitro, the cell cycles and the proliferation of astrocytes in different time points after ischemia and anoxia were studied by flow cytometry and BrdU labeling and the expression of GFAP and cyclin D1 was detected by the fluorescence immunochemistry. After ischemia and anoxia in vitro, the astrocytes in S phase were significantly increased as compared with those in the normal group and the proliferating ability of the astrocytes was highest 6 h after the treatment as revealed by BrdU pulse labeling, but the astrocytes in S phase and proliferating ability were decreased after 6 h. At the early stages of ischemia and anoxia, the positive staining intensity of GFAP was increased, peaked at 6th h, while 12 h after the ischemia and anoxia, the positive staining intensity of GFAP became weak, and the expression of cyclin D1 was gradually increased after the ischemic and anoxic damage. It is concluded that astrocytes are activated to proliferate and enter new cycle events by ischemia and anoxia, and cyclin D1 is implicated in the proliferation and repair of astrocytes. The cell cycle events are closely associated with the proliferation and activation of astrocytes.
Animals, Newborn ; Astrocytes/*cytology ; Cell Cycle ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Cyclin D1/biosynthesis ; Cyclin D1/genetics ; Glial Fibrillary Acidic Protein/*biosynthesis ; Glial Fibrillary Acidic Protein/genetics ; Rats, Wistar

OBJECTIVEHighland natives adapt well to the hypoxic environment at high altitude (HA). Several genes have been reported to be linked to HA adaptation. Previous studies showed that the endothelial ni- tric oxide synthase (ENOS) G894T polymorphism contributed to the physiology and pathophysiology of hu- mans at HA by regulating the production of NO. In this meta-analysis, we evaluate the association between the ENOS G894T polymorphism and HA adaptation through analyzing the published data.

METHODSWe searched all relevant literature about the ENOS G894T polymorphism and HA adaptation in PubMed, Med- line, and Embase before Step 2015. A random-effects model was applied (Revman 5.0), and study quality was assessed in duplicate. Six studies with 634 HA native cases and 621 low-altitude controls were included in this meta-analysis.

RESULTSFrom the results, we observed that the wild-type allele G was significantly overrepresented in the HA groups (OR = 1.85; 95% Cl, 1.47-2.33; P < 0.0001). In addition, the GG genotype was significantly associated with HA adaptation (OR = 1.99; 95% Cl, 1.54-2.57; P < 0.0001).

CONCLUSIONOur results showed that in 894 G allele carriers, the GG genotype might be a beneficial factor for HA adaptation through enhancing the level of NO. However, more studies were needed to confirm our findings due to the limited sample size.

Adaptation, Physiological ; genetics ; Altitude ; Genotype ; Humans ; Nitric Oxide Synthase Type III ; genetics ; Polymorphism, Single Nucleotide

OBJECTIVETo study the effect of substrate stiffness on proliferation, migration of fibroblast and integrin β(1) expression in fibroblast.

METHODSFibroblasts were inoculated on silicon substrate with stiffness of (16.2 ± 0.5), (19.8 ± 1.1), and (200.1 ± 2.6) kPa. After being cultured for 5 days or 6 days, cells were counted and cell proliferative activities (recorded as absorbance value) were assessed with methyl thiazolyl blue (MTT). After being cultured for 3 days, cell cycle was detected and proliferation index (PI) was calculated. The cell scratch test was used for determination of cell migration rate on post scratch day (PSD) 0 (the day of scratch), 1, 2, and 3. After being cultured for 2 days, the expression of integrin β(1) was determined by flow cytometry with fluorescence. Data were processed with one-way analysis of variance.

RESULTS(1) The proliferative speed and proliferative activity of fibroblasts were all increased along with the increase in substrate stiffness. PI of fibroblasts inoculated on silicon substrate with stiffness of (16.2 ± 0.5), (19.8 ± 1.1), and (200.1 ± 2.6) kPa was respectively 24.8%, 27.4%, 32.4%. On PSD 2, migration rate of fibroblasts inoculated on silicon substrate with stiffness of (19.8 ± 1.1) and (200.1 ± 2.6) kPa was respectively (91.4 ± 5.1)%, (100.0 ± 1.3)%, which were higher than that of fibroblasts inoculated on silicon substrate with stiffness of (16.2 ± 0.5) kPa [(55.8 ± 6.8)%, with F value respectively 3.5, 4.0, P values all below 0.01]. (3) The expression rate of integrin β(1) in fibroblasts inoculated on silicon substrate with stiffness of (16.2 ± 0.5) kPa was the lowest (43.22%), and that in fibroblast inoculated on silicon substrate with stiffness of (200.1 ± 2.6) kPa was the highest (81.26%).

CONCLUSIONSSubstrate stiffness may have a great effect on proliferation and migration of fibroblast during the process of wound healing and scar formation, which can be related to regulation of integrin β(1) expression.

Cell Movement ; Cell Proliferation ; Cells, Cultured ; Fibroblasts ; cytology ; metabolism ; pathology ; Humans ; Integrin beta1 ; metabolism ; Mechanical Phenomena ; Silicon

Objective To explore the influence of different administration time on antidepressant effect of seven clinical common antidepressants. Methods Male mice were randomly divided into eight groups:venlafaxine (75 mg/kg), sertraline (20 mg/kg), fluoxetine (20 mg/kg), doxepin (15 mg/kg), mirtazapine (15 mg/kg), citalopram (40 mg/kg), trazodo?ne (50 mg/kg) and control (saline) groups. Each group contained 36 mice. Drugs were administered to 6 mice per group 30 min before forced swimming test at the 6 time points (9:00, 13:00 and 17:00 as light phase and 21:00, 1:00 and 5:00 as dark phase). Forced swimming test was applied to determine the influence of dosing time on anti-immobility effect of seven antidepressants at each time point. Results Immobility time in venlafaxine group and sertraline group significant?ly decreased compared with that of control group at all time points(all P<0.05). Moreover, anti-immobility effects of ven?lafaxine, fluoxetine, mirtazapine and doxepin were better during the dark phase than during the light phase (all P<0.05). In addition, immobility time in sertraline group decreased at the late part of dark phase (5:00) and the early part of light phase (9:00) compared with other phases (P<0.05). Conclusions Most antidepressants show 24-h rhythm dependent an?ti-immobility effects, but rhythmic patterns are not completely consistent among different antidepressants. Further study is needed to explore the chronopharmacological mechanism and clinical applications of these antidepressants.

Objective To investigate the specific expression of endoplasmic reticulum stress-related GRP78 and CHOP in podocyte of MKR mice with diabetic nephropathy (DN);To discuss the intervention effect of Zuogui Jiangtang Yishen Decoction (ZGJTYS). Methods 8-week old MKR mice were randomly divided into five groups:blank group, model group, ZGJTYS group, 4-Phenylbutyric acid group, gliquidone+benazepril group, and normal control group consisting of wild type C57BC/6 mice, 10 mice per group. All mice from model group and each treatment group received high-fat diet feed and unilateral nephrectomy to make the DN model. Mice from treatment groups received medicine intervention for four weeks. The levels of FBG were detected by electrochemical detection method, and the UmAlb was detected by ELISA. The expressions of GRP78, CHOP mRNA, and protein were detected by RT-PCR and Western blot. The morphological structure changes of the podocytes were observed by transmission electron microscopes. Results Compared with blank group, FBG and UmAlb in the model group increased significantly (P<0.01);the expressions of GRP78 mRNA and protein markedly decreased (P<0.01);the expressions of CHOP mRNA and protein increased (P<0.01). Compared with the model group, FBG and UmAlb in each treatment group significantly decreased (P<0.01);the expressions of GRP78 mRNA and protein in the ZGJTYS group and 4-Phenylbutyric acid group increased (P<0.01), the expressions of CHOP mRNA and protein decreased (P<0.01). The renal pathological lesions of each treatment group were significantly improved compared with model group. Conclusion ZGJTYS Decoction can effectively improve the damaged podocyte induced by endoplasmic reticulum stress, delay DN progress.

DNA Replication ; physiology ; Epstein-Barr Virus Infections ; virology ; Herpesvirus 4, Human ; genetics ; metabolism ; physiology ; Humans

Objective To observe the effects of ICB orthopedic sole combined with rehabilitation training on balance and walking function in stroke survivors.Methods Thirty hemiplegic stroke patients were recruited and divided into a study group (n =15) and a control group (n =15) by using a random number table.Both groups took exercises based on the principles of a motor relearning program and conducted core stability control training,and the study group additionally undertook hemiplegic lower extremity weight-bearing exercises and walking with ICB orthopedic sole used in daily living.The program was administered 20 min twice per day for 4 weeks.All patients were evaluated with Rest Calcaneus Standing Position (RCSP),Malleolar Position (MP),Forefoot Position (FP),Berg Balance Scale (BBS),l0 m Maximum Walking Speed (MWS) and walking section of Motor Assessment Scale (MAS)before and after the program.Results After 4 weeks of intervention,all the measurements except the FP in both groups improved significantly,and significant differences were observed between the two groups.After 4 weeks of training,the average RCSP (1.78 ± 0.32) ° and MP (13.33 ± 2.51)° were improved significantly compared to those of the control group [(2.58 ± 0.59) ° and (12.45 ± 3.31) °,respectively].Moreover,the average BBS,MAS and MWS improved significantly compared to the control group.Conclusions ICB orthopedic sole combined with rehabilitation training can improve the weight-bearing,balance and ambulation abilities of stroke survivors.

Objective To verify the methodology for auditing dosimetric parameters in reference and non-reference conditions with thermoluminescent dosimeters (TLDs).Methods Under reference and non-reference conditions,the established TLD methods were used to observe the absorbed dose variations with depth,SSD,field size and 45 wedges for 10 photon beams at 5 hospitals.Dosimetric parameters,including doses at Dmax points in axis,on 5 electron beams of 9 MeV were measured.The measurement results were compared between the TLDs and plane parallel ionization chambers.Results For 6 MV photon beams,the relative deviation of between finger ionization chamber method and TLD chips was in the range of-1.7％ to 5.4％ under on-axis non-reference conditions,and-6.3％ to-0.6％ under off-axis non-reference conditions,respectively,all within the range of ≤ ± 7％ as required by the IAEA.The relative deviation between plane parallel chamber and TLD method was-2.3％ to 3.7％,within ± 5％ as required by the IAEA.Conclusions It is convenient and feasible to use TLD method for quality audits of dosimetric parameters in radiotherapy.

Background Dyslipidemia is one of the major causes of age-related macular degeneration (AMD).At present,the study of the preventive and treating methods of A MD is still a hot spot.Objective The purpose of this study was to observe the effect of tonifying the spleen and promoting blood circulation on the retina and Bruch membrane in apolipoprotein E-deficient (ApoE-/-) mice with dyslipidemia.Methods Thirty-six ApoE-/-mice aged 2 months were randomly divided into the normal diet group,high fat diet group and medicine group.A diet with a higher content of fat was given for 5 consecutive months to the mice of the high fat diet group and medicine group,and in the last month,a concoction that tonifies the spleen and promotes blood circulation was gavagely administered in the medicine group,and an equivalent volumes of normal saline solution was administered in the same way in the normal diet group and high fat diet group.Total plasma cholesterol (TC),low density lipoprotein (LDL)and triglyceride (TG) were detected by (ELISA? Name of assay?) using the 7170 Hitachi automatic biochemical analyzer,and morphological changes of the retina and Bruch membrane were examined by light microscopy and transmission electron microscopy.The number of outer nuclear layer (ONL) cells,retinal pigment epithelial (RPE) cells and the thickness of the Bruch membrane were examined by semi-quantitative histopathology with the Mias 2000 Imaging Analyzer System.Results The concentrations of TC,LDL and TG were (6.47 ±0.49) mmol/L,(1.46 ±0.10)mmol/L and (0.62 ±0.21) mmol/L,respectively,in 7-month-old mice of the medicine group,showing a significant reduction in comparison with (10.53 ±0.30) mmol/L,(1.90±0.13) mmol/L,(1.15±0.29) mmol/L of the high fat diet group,and (9.63 ± 0.18) mmol/L,(1.12 ± 0.15) mmol/L,(0.88 ± 0.21) mmol/L in the normal diet group (P＜0.05-0.01).The disorder and atrophy of ONL and RPE cells,divergence of fiber of the Bruch membranes were found in both the high fat diet group and normal diet control group under the light microscope,and drusen formed in some of the mice in the high fat diet group.However,ONL and RPE were well organized in the medicine group.The cell numbers in the ONL and RPE layer in the 7-month-old mice were (23 124.00±755.18) and (10.75±0.59),respectively,in the medicine group,(19 107.00 ± 1436.82) and (8.55 ± 1.11),respectively,in the high fat diet group,(21 663.00± 1073.27) and (9.75 ±0.58),respectively,in the normal diet group,with significant differences among them (P＜0.05-0.001).Thickness of the Bruch membrane in the medicine group extensively reduced in high fat diet group and normal diet control group (P＜0.01).The ultrastructures of the RPE and Bruch membrane were much more improved in the mdedicine group.Conclusions Tonifying the spleen and promoting blood circulation can attenuate hyperlipemia in ApoE-/-mouse;furthermore,it lessens the pathological abnormalities in the ONL,RPE and Bruch membrane.

RhoA, a small GTPase, is involved in a wide array of cellular functions in the central nervous system, such as cell motility, cytoskeleton rearrangement, transcriptional regulation, phagocytosis and cell growth. It is not known how spinal cord injury (SCI) affects the expression of RhoA in different nerve cells. In the present study, we investigated the changes of RhoA expression in remote areas of the injury at the 3rd, 7th and 30th day after SCI, which was established by T10 contusion method. Moreover, we examine its expression profile in neurons, astrocytes and microglia. RhoA was found to be weakly expressed in these nerve cells in normal spinal cord. Western blotting showed that, after SCI, the total RhoA expression was up-regulated, and the RhoA expression was increased and peaked at the 7th day. Double immunostaining revealed specific and temporal expression patterns of RhoA in different nerve cells. The expression of RhoA in neurons started to increase at day 3, peaked at day 7 and then decreased slightly at day 30. Expression of RhoA in astrocytes increased moderately after SCI and peaked at day 7. There was no obvious change in RhoA expression in microglia after SCI in remote areas. This study demonstrated that, after SCI, RhoA expression exhibited different patterns with different nerve cells of spinal cord. RhoA expression patterns also changed with time after SCI, and among different nerve cells in the injured spinal cord. These findings can help us better understand the roles of RhoA in SCI.