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Isolation and Adherent Culture of Neural Stem Cells from Mouse Embryo

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2017;46(3):312-316,340. doi:10.3870/j.issn.1672-0741.2017.03.015

Objective To develop a modified method of adherent culturing NSCs to replace the traditional suspension culturing method.Methods Neural stem cells(NSCs)were isolated from brain of fetal mouse at 15.5-day fetal age.In culture bottle which had been pre-coated with poly-L-ornithine hydrochloride/fibronectin(PO/FN),the cells were adherently cultured and passaged.The cell morphology was observed under inverted microscope.NSCs and their differentiated cells were identified by immunofluorescence.Results NSCs obtained in this study were successfully adherently cultured and expressed specific markers.Conclusion NSCs are successfully adherently cultured in this study.As compared with the traditional suspension culturing method,adherent culturing is simpler and has lower wastage of passage.It is easy to observe cell morphology and differentiation by this method.It is also helpful to culturing and storage of NSCs.

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Evaluation of Intervention on AIDS High-risk Sexual Behavior among Young Students in Jilin MSM Population

Yong CAI ; Jianguo QIAO ; Dianwei SUN

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2017;46(3):346-351,367. doi:10.3870/j.issn.1672-0741.2017.03.022

Objective To evaluate effect of intervention on acquired immune deficiency syndrome(AIDS) high-risk sexual behavior of males who have sex with males(MSM)population in Jilin,and to provide a basis for establishing apropriate intervention strategy for this population.Methods A cross-sectional survey was carried out on the MSM population of young students with the method of classification.Comprehensive intervention strategy included health education via new media,behavioral intervention,voluntary conseling and testing,peer education,condom promotion and standardized services for clinic of sexually transmitted diseases(STDs)and other comprehensive interventions.Before and after intervention,300 MSM were enrolled,and their demographic characteristics,knowledge of AIDS prevention and control,high risk behaviors and intervention measures were investigated and analyzed.Results Before and after the intervention,the awareness rate of AIDS related knowledge was 67.67％ and 94.33％,respectively,the condom usage rate was 23.57％ and 11.79％ in the most recent and the last 6 months before the intervention,and it was 76.21％ and 62.10 ％ after the intervention,respectively.The proportion for those who used condom every time in heterosexual sex behavior was increased from 23.53％ (before intervention) to 69.60 ％ (after intervention),and the condom usage rate in the last 6 months was increased from 15.44％ to 66.40％.Proportion of the MSM who had STDs related symptoms was decreased from 21.33％ to 7.67％ after the intervention.Conclusion After the comprehensive intervention,the awareness rate of AIDS related knowledge among MSM in Jilin is significantly increased,the incidence of highrisk sexual behavior is decreased and the condom usage rate is improved.Intervention by use of new media is low-cost,easy to operate,and can be used as a good supplementary strategy for traditional behavioral intervention.

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Effect of Treatment of a Non-surgical Spinal Decompression System on Neck Muscle Surface Electromyography of Patients with Cervical Spondylotic Radiculopathy

Xiaoxiao XU ; Chuhuai WANG

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2017;46(3):336-340. doi:10.3870/j.issn.1672-0741.2017.03.020

Objective To explore effect of treatment of a non-surgical spinal decompression system(SDS) on neck muscle surface electromyography(EMG)of patients with cervical spondylotic radiculopathy(CSR).Methods Sixty patients with CSR in Department of Rehabilitation Medicine at The First Affiliated Hospital of Sun Yat-sen University in China between February 2014 and February 2016 were recruited and randomly divided into SDS group and control group,with 30 patients for each group.The SDS group and control group were treated with the SDS and general traction system for cervical traction respectively.Surface EMG telemeter was used to measure the surface EMG signals of the affected side cervical erector spinae muscle and sternocleidomastoid with the score of averaged EMG value(AEMG)and the median frequency(MF)as surface electromyogram signal evaluation indexes,and VAS and NDI as therapeutic effect evaluation indexes after a course of treatment.The data before and after the treatment were statistiacaly analyzed.Results During the first traction with tradtional traction or SDS,AEMG values of the cervical paraspinal muscle and sternocleidomastoi were significantly decreased and MF values significantly increased as compared with those before the traction (all P＜0.05).After a course of treatment,AEMG values of affected side cervical erector spinae muscle and sternocleidomastoi of SDS group were significantly decreased as compared with those before the treatment,and they were significantly lower in SDS group than in ordinary group (all P＜0.05);and MF values of affected side cervical erector spinae muscle and sternocleidomastoi in SDS group were significantly increased as compared with those before the treatment,and they were significantly higher in the SDS group than in the ordinary group (all P＜0.05).VAS and NDI scores of the two groups after a course of treatment were significantly lower than those before the treatment (P＜0.05),and VAS and NDI scores were significantly lower in the SDS group than in the ordinary group (P＜0.05).Conclusion SDS traction is superior to ordinary traction in relieving pain,improving function,relaxing neck muscles and alleviating muscle fatigue.

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Expression of PP2CE in Human Hepatocellular Carcinoma Tissues and the Clinical Significance

Weihua XU ; Huanjun HUANG

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2017;46(3):327-331. doi:10.3870/j.issn.1672-0741.2017.03.018

Objective To investigate expression of PP2CE in human hepatocellular carcinoma(HCC)tissues and the clinical pathological significance,and test its sub-cellular localization in liver cells and HCC cells.Methods A total of 50 samples of HCC tissues and their para-carcinoma tissues were collected in Tongji Hospital(Wuhan).PP2CE mRNA and protein levels were detected in HCC tissues and their para-carcinoma tissues by qRT-PCR and immunohistochemistry method.Immunofluorescent assay was performed to test the sub-cellular localization of PP2CE in the cells.Results PP2CE mRNA and protein expression levels were significantly decreased in HCC tissues as compared with those in para-carcinoma tissues(both P＜0.05).Immunohistochemistry analysis found that PP2CE expression was associated with tumor size,differentiated degree,portal vein invasion and clinical stage(both P＜0.05).Immunofluorescent assay suggested that PP2CE was mainly located in the cytoplasm and perinuclear area.Conclusion PP2CE is reduced in HCC and may be implicated in the development and progression of HCC.

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Anti-inflammatory and Anti-angiogenic Effect of miR-15a on Diabetic Retinopathy

Jia SHI ; Chen TANG ; Hongbin LV

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2017;46(4):443-448. doi:10.3870/j.issn.1672-0741.2017.04.014

Objective To explore the anti-inflammatory and anti-angiogenic effect of miR-15a in diabetic retinopathy(DR).Methods High glucose model(HG)was established by treatment with 25 mmol/L glucose in human retinal pigment epithelial cell line HARPE-19,and HARPE-19 cells treated with 5 mmol/L glucose served as control(LG).HG group and LG group were divided into 4 subgroups:miR-15a mimic group,NC-mimic group,miR-15a inhibitor group and NC-inhibitor group.Cell proliferation ability was detected by MTT proliferation assay.Cell migration ability was detected by cell scratch assay.The relationship between miR-15a and ASM was identified by 3′UTR luciferase reporter assay.miR-15a,ASM,VEGF,IL-1β,IL-6,TNF-α mRNA expression was detected by RT-PCR method.Results In NC-mimic cells and NC-inhibitor cells,the expression of miR-15a in HG group was significantly lower than that in LG group(all P<0.05).The expression level of ASM mRNA in miR-15a mimic group was significantly lower than that in NC-mimic group (P<0.05),the expression level of ASM mRNA in miR-15a inhibitor group was significantly higher than that in NC-inhibitor group (P<0.05).The cell scratch width in miR-15a mimic group was significantly higher than that in NC-mimic group(P<0.05).The cell scratch width in miR-15a inhibitor group was significantly lower than that in NC-inhibitor group(P<0.05).The absorbance value in miR-15a mimic group was significantly higher than that in NC-mimic group(P<0.05),the absorbance value in miR-15a inhibitor group was significantly lower than that in NC-inhibitor group(P<0.05).In NC-mimic cells and NC-inhibitor cells,the mRNA expression levels of VEGF,IL-1β,IL-6 and TNF-α in HG group was significantly higher than those in LG group(all P<0.05).Conclusion miR-15a downexpression induces inflammatory reaction and angiogenesis in retinal cells.miR-15a is involved in the pathogenesis of DR by dual regulation of inflammation and angiogenesis.

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Carnosic Acid Cooperates with Tamoxifen to Induce Apoptosis of Breast Cancer Cells through Caspase-3 Signaling Pathway

Wenyu HE ; Changjing YUAN

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2017;46(4):404-409. doi:10.3870/j.issn.1672-0741.2017.04.007

Objective To explore the effect of carnosic acid combined with tamoxifen on breast cancer cells and the mechanism.Methods Breast cancer cell lines MCF-7,T47D were divided into 4 groups:PBS was given to control group,carnosic acid(CA)was added to CA group,tamoxifen(TAM)was added to TAM group,and CA+TAM group was treated with carnosic acid and tamoxifen.The proliferation ability was measured by MTT assay,the colony formation capacity was measured by colony formation assay,the invasion ability was measured by Transwell chamber assay,the migration ability was measured by wound-healing assay,the expression of Caspase-8,Caspase-3 and PARP protein were measured by Western blotting.Results CA,TAM and CA+TAM suppressed breast cancer cell proliferation with a dose-dependent manner.Compared with CA group and TAM group,the proliferation inhibitory rate of CA+TAM group was significantly increased(P<0.05).In MCF-7 and T47D cell lines,compared to control group,the rate of colony fromation,number of invasive cells and cell migration rate of CA,TAM,CA+TAM groups were significantly decreased(all P<0.05).These indexes mentioned above in CA+TAM group were significantly lower than those in CA group and TAM group(all P<0.05),but there were no significant differences between CA group and TAM group (all P>0.05).In two breast cancer cell strains,the expression of cleaved Caspase-8 protein between control,CA,TAM and CA+TAM groups was not significantly different(P>0.05).Compared to control group,the expression levels of cleaved caspase-3,cleaved PARP protein in CA,TAM and CA+TAM groups were significantly increased(P<0.05),and these protein levels were significantly higher in CA+TAM group than in CA and TAM group(all P<0.05).But there were no significant differences between CA group and TAM group(all P>0.05).Conclusion Carnosic acid combined with tamoxifen can suppress the proliferation,invasion and migration of breast cancer cells,and its mechanism may be related to the activation of Caspase-3/PARP signaling pathway.

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Melatonin Ameliorates Tau Hyperphosphorylation of Mice with Exposure to Lead(Pb)at Early Development Stage by Mediating ER Stress in Hippocampus

Xiaofeng LIU ; Juan CHEN ; Qishui OU

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2017;46(4):392-396. doi:10.3870/j.issn.1672-0741.2017.04.005

Objective To investigate the effect of melatonin(MT)on tau hyperphosphorylation in hippocampus of mice with exposure to lead(Pb)at early development stage and its possible mechanism.Methods Healthy C57BL/6 mice were randomly divided into four groups:control group(received normal water),lead exposure group(exposed to 0.2% Pb acetate from postnatal day 1(PND 1)to PND 21 through drinking water),MT group(received 50 mg/mL MT through drinking water since 12-month-old for 3 months)and MT combined with lead exposure group(exposed to 0.2% Pb acetate from PND 1 to PND 21 and then given 50 mg/mL MT since 12-month-old for 3 months through drinking water).The Pb levels in the blood and hippocampus were determined by graphite furnace atomic absorption spectrometry.Morris water maze was used to detect the spatial learning and memory.The phosphorylation level of tau and the protein level of GRP78 and CHOP in hippocampus were detected by Western blotting.Results Compared with the control group,the Pb levels in the blood and hippocampus were significantly increased in lead exposure group(P<0.05),while MT treatment did not affect the Pb levels both in the blood and hippocampus.In the lead exposure group,the phosphorylation level of tau in the hippocampus was significantly increased compared with control group(P<0.01),and after treatment with MT,the phosphorylation level of tau in MT combined with lead exposure group was significantly decreased compared with the lead exposure group(P<0.05).Furthermore,the expression of GRP78 and CHOP in the hippocampus was significantly higher in lead exposure group than in control group(P<0.05),and after treatment with MT,the expression levels of GRP78 and CHOP were lower than those of the control group.Conclusion MT can ameliorate the phosphorylation level of tau in the hippocampus and defects of learning and memory in C57BL/6 mice with exposure to lead at early development stage,and this may be related with the modulation of endoplasmic reticulum stress.

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Expression and Clinical Significance of PPARγ in Bladder Urothelial Cancer

Guang SHAN ; Huijun QIAN ; Yue XIA

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2015;(4):468-471,487. doi:10.3870/j.issn.1672-0741.2015.04.021

Objective To investigate the expression and clinical significance of peroxisome proliferator activated receptor γ(PPARγ)in bladder urothelial cancer tissues.Methods Parafflin‐embeded specimen of bladder urothelial cancer tissues from 50 cases and normal tissues near the bladder urothelial cancer from 5 cases were harvested from the Pathology Department of the Renmin Hospital of Wuhan University between 2006 and 2009.Those cases had complete pathological and clinical data.The ex‐pression of PPARγ was detected by immunohistochemical SP method.Quantitative analysis of the PPARγ was measured by high definition pathological graphics context report system (HPIAS‐1000).One‐way analysis of variance and SNK (q)tests were used to analyze the mean density and the positive area rate of the immunohistochemical results.All data were processed by SPSS 13.0.Results The expression of PPARγwas significantly higher in bladder urothelial cancer tissues than in para‐carcinoma tis‐sues(P<0.05).Correlation between expression of PPARγ with TNM stag of bladder urothelial cancer was as follows :Positive rate of PPARγin the tissues with primary tumor size ≥3 cm was 72.4% ,significantly higher than 33.3% in the tissues with tumor size <3 cm(P<0.05);positive rate of PPARγin the cases with lymph node metastasis was 72.7% ,significantly higher than 46.4% in the cases without lymph node metastasis(P<0.05);positive rate of PPARγin patients at stage T3‐4 group was 75.0% ,significantly higher than 41.9% and 45.5% in patients at clinical stage T1 and T2(P<0.05);positive rate of PPARγin patients with poor differentiation was 68.2% ,significantly higher than 42.9% in patients with high or middle differentiation group(P<0.05).Conclusion PPARγ plays an important regulating role in the onset and progress of bladder urothelial cancer ;PPARγexpression level was correlated with primary tumor size ,pathological types and differentiation degree ,lymph node me‐tastasis and clinical stage.This result suggested that PPARγ was closely correlated to metastasis of bladder urothelial cancer.

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Noninvasive Detection of Liver Fibrosis by Fibroscan in Asymptomatic Carriers with Chronic Hepatitis B Virus

Hongfang DING ; Ke MA ; Qin NING

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2015;(5):572-574,590. doi:10.3870/j.issn.1672-0741.2015.05.017

Objective To investigate the clinical application of Fibroscan to noninvasive detection of liver fibrosis in asymp‐tomatic carriers(ASCs)with chronic hepatitis B virus(HBV).Methods Clinical data of 195 chronic HBV ASCs were retrospec‐tively analyzed ,including liver function ,HBV DNA quantitation ,liver stiffness(LS) ,color Doppler ultrasound results of the liver and spleen and histopathological findings of the liver.LS was evaluated by transient elastography(Fibroscan).Liver biopsy was performed in 80 carriers for histological examination.Results The LS value was(6.22 ± 2.50)kPa in 195 cases.There was no significant difference in the LS value between HBV DNA <500 copies/mL and HBV DNA≥500 copies/mL groups.The LS value in patients with HBV carrying duration>15 years was significantly higher than that in patients with HBV carrying dura‐tion≤5 years(P<0.01).It was (6.72 ± 2.33) kPa in patients(n=49)with liver tissue inflammation and necrosis

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Analysis of the Sexual Behavior and the Influential Factors of AIDS Infection among Out-of-school Adolescent MSM in Chongqing

Cuifang LONG ; Jianping NIE ; Yiping YAN

Acta Medicinae Universitatis Scientiae et Technologiae Huazhong.2015;(5):603-607,615. doi:10.3870/j.issn.1672-0741.2015.05.025

Objective To examine the current status of the sexual behavior among out‐of‐school adolescent men who have sex with men(MSM)and analyze the influential factors of AIDS infection in order to provide countermeasures.Methods The software SPSS 16.0 was used for cross table analysis and chi square test of questionnaire data and HIV detection data.Results The HIV infection rate was 21.36%.The household registration ,the only child ,living state ,coming out of the closet ,fixed sex‐ual partner ,the way they know sexual partners had no significant effects on HIV infection(P>0.05).Having multiple sexual partners(P<0.05) ,unfamiliar with sexual partners(P<0.05) ,having sex with network friends(P<0.05) ,no condom use in sexual life(P<0.01) ,having no regular HIV testing(P<0.01)were significantly related to HIV infection.Conclusion Reduc‐ing the sexual behavior among out‐of‐school adolescent MSM ,carrying out HIV counseling and testing ,promoting the use of condoms contribute to the reduction of HIV infection.

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