Purpose To observe the therapeutic effect of acupuncture therapy plus Tuina therapy in the treatment of simple obesity. Methods Ninety-eight cases of the patients were randomly divided into the two groups and 60 cases in the treatment group were treated with acupuncture therapy plus Tuina therapy, and 38 cases in the control group were treated with single acupuncture therapy. Results The effective rate was 100% in the treatment group and was 71.0% in the control group, P ＜ 0. 01. Conclusion Acupuncture therapy plus Tuina therapy is better than single acupuncture therapy in the treatment of simple obesity.

Objective To observe the effects of ginkgolide B (GB) on mRNA expression of foxgl and proliferation of cells in brain tissue of newborn rats with hypoxic-ischemic brain damage (HIBD).Methods A total of 128 clean 7-day-old healthy SD rats were randomly divided into sham operation group,the model group,the low-GB dose and the high-GB dose treatment groups.Classic Rice method were used to establish HIBD models in the latter 3 groups.Four hours after operation,and GB in dose of 5 mg/kg and 10 mg/kg was given to rats in the low and the high dose treatment groups by intraperitoneal injection postoperatively,once a day for 5 days,while sham operation and model groups were treated with equal physiological saline.All groups were respectively sacrificed on 3 d,7 d,14 d,28 d respectively.Quantitative real-time fluorescent polymerase chain reaction was employed to detect expression of Foxg1 gene.Then the number of 5-bromodeoxyuridine positive cell in subgranular zone was investigated by immunolluorescent stairning.Results The Foxg1 mRNA expression was observed 3 days after HIBD,peaked on 7th day,and then declined gradually; the levels of Foxg1 mRNA in the 2 treatment groups were higher than that of the HIBD group (all P ＜ 0.01) ; The expression of Foxgl at 7 d,14 d,28 d,in high-dose group were higher than those in the low-dose group (all P ＜ 0.01).The number of 5-bromodeoxyuridine positive cell was increased after HIBD,and the levels in the low-and the high-dose treatment groups were all higher than that of the model group (all P ＜ 0.05) ; the number of positive cell in high-dose treatment groups were higher than that in the low-dose treatment groups (P ＜ 0.05).Conclusions GB can promote the expression of Foxg1 gene and improve the proliferation of cells in Brain tissue after HIBD,which shows more significant efficacy in high-dose group than in low-dose group.

Objective To detect the COMP and aggrecan content in serum of patients with KOA and explore its correlative factors.Methods COMP and aggrecan in serum of 100 patients with knee-osteoarthritis(KOA group)and 30 healthy volunteers (control group)were measured by ELISA.Correlative factors for COMP and aggrecan levels were tested by linear regression analy-sis.Results COMP and aggrecan levels in KOA group were significantly higher than that in control group (P < 0.05 ).Serum COMP and aggrecan levels were positively correlated with age,body mass index(BMI),WOMAC scores and X-ray K-L grade(P <0.05).Conclusion Serum COMP and aggrecan are useful markers in the diagnosis,which their level in KOA are higher than the normal people.Moreover,these rise of levels positively correlated with clinical disease parameters and radiological joint damage.

Objective:To investigate the effect of ethyl acetate extract from Cortex periplocae (CPEAE) on apoptosis of human esophageal carcinoma cell line TE-13 and to elucidate its mechanism. Methods:Inhibitory effect of CPEAE on TE-13 proliferation was tested by MTT assay. The morphological changes of cell apoptosis were observed by Giemsa staining and transmission electron microscopy. Cell cycle and apoptotic ratio were tested by flow cytometry (FCM). The protein expression of CDK4 was observed by Western blotting.Results:CPEAE inhibited proliferation of TE-13 cells in a concentration-dependent and time-dependent manner, and its IC_(50) value was (2.443±0.005) μg/mL at 48 h (P<0.05). The characteristic morphological changes of apoptosis were observed in TE-13 cells after treatment with CPEAE under transmission microscope. A typical subdiploid peak was detected by flow cytometry. CPEAE decreased the expression of gene CDK4 in TE-13 cells. Conclusion:CPEAE can induce apoptosis of TE-13 cells. The effect is related with down-regulation of CDK4 expression.

ObjectiveTo screen mimetic HIV-1 neutralizing epitopes from plasma with high level neutralizing antibody,and to provide useful information for further study of the interaction between antigen and antibody.MethodsIn order to gain neutralizing antibody recognized mimotopes, we detected neutralizing antibodieslevelsof 11HIV-1infectedslowprogressorsbyPBMC-basedneutralization assays.High-titer HIV-neutralizing antibodies from plasma of SPs was used as the ligand for biopanning by phage-displayed random peptide library.Positive phage clones was evaluated by ELISA,sequenced,and analyzed for homology to HIV-1 env by local BLAST to deduce the neutralizing peptide.ResultsTwenty-two clones were obtained consistent with requirement through three rounds biopanning.After comparison analysis,twelve clones include C8 were obtained as mimotopes of neutralizing antibody,C40 located in gp41Ⅱ cluster with the highest titer by inhibition ratio may be as neutralizing epitope.Conclusion By the use of IgG antibodies from SPs to screen the phage random polypeptide library,one can acquire multiple phage mimetic peptides of HIV related antigen epitope.(Chin J Lab Med,2012,35:838-842 )

Objective To explore the polymorphism of nef gene and conservation level of functionally important domains of nef as well as their influences on HIV-1 disease progression of HIV-1 B'infected individuals in northern China.Methods 30 long term nonprogressors(LTNPs)and 42 typical progressors (TPs)were selected.Provirus DNA was extracted from whole blood sample.The full nef gene was amplified by nested-PCR.PCR product was sequenced directly after purification.Phylogenetic analysis and amino acid sequence mutation was applied on nef sequences to explore the differences between LTNPs and TPs.Results At position 15,the S15R/K/N substitution was detected.The frequency of TPs(64.29%)wsa higher than LTNPs(33.33%,P<0.01,0R=3.60);R21K/E/H/I.Q,TPs and LTNPs mutation frequency was 59.52%and 93.33%(P<0.005,OR=0.11);At position 39,K39R/E/N was only detected in TPs(23.81%,P<0.005).Conclusion No significant deletion or defect associated with disease progression wsa detected in nef gene of HIV-1 B'.But it suggested that K/E/H/I/Q mutation at 21 st amino acid of nef associated the disease nonprogression.R/K/N at 15 th amino acid of nef and R/E/N mutation at 39th amino acid of nef associated the disease progression in.HIV-1 B'.All domaias of nef amino acids sequences were comparatively conservative.

To explore the effect of estrogen on pathogenesis of osteoporosis in women with maintenance hemodialysis. One hundred and twenty women aged 18-45 years had been undergoing maintenance hemodialysis for ≥ 3 months were included. Of them ,60 women without osteoporosis served as control group and the other 60 women with osteoporosis as observation group. Serum concentrations of estradiol, tumour necrosis factor-α (TNF-α), parathyroid hormone (PTH), and calcium were determined, meanwhile bone mineral density (BMD)was measured by quantitative computed tomography. Serum estradiol levels in the observation group were lower while TNF-α level were higher than those in control group (all P<0. 05). PTH and calcium levels were not significantly different (P= 0.567 and P = 0. 588). In the observation group, linear correlation analysis revealed positive correlation (r = 0. 865 ,P<0. 01)between estradioi and BMD,while multiple linear regression analysis showed that serum estradiol and calcium levels were positively correlated with BMD, and the concentrations of TNF-α and PTH were negatively correlated with BMD (F= 140.32 ,P<0.01). Estradiol levels were found to have greater effect on BMD(t=5. 386, P<0. 01). Lowered serum concentration of estradiol in women with maintenance hemedialysis seems to be a major factor related to osteoporosis,it accelerates the pathogenesis of osteoporosis by modulating TNF-α.

AIM:To explore a novel method to isolate human nucleus pulposus mesenchymal stem cells (hNP-MSCs) in vitro and to identify their biological characteristics.METHODS:The explant culture method was employed to isolate hNP-MSCs from nucleus pulposus tissue obtained by percutaneous endoscopic lumbar discectomy (PELD).The isolated cells were passaged for purification and cultured in vitro followed by morphological observation.The cell proliferation ability was detected by CCK-8 assay.Growth curves of the cells were drawn and surface antigens were detected by flow cytometry.The cells at the 3rd~6th passages were induced for adipogenic, osteogenic and chondrogenic differentiation, and examined by oil red O staining, alizarin red staining and Alcian blue staining.RESULTS:The cells with self-renewal were obtained from nucleus pulposus tissue obtained by PELD.The results of flow cytometry analysis revealed that the cells were positive for CD29, CD44, CD90, CD73 and CD105, but negative for CD34 and CD45.The proliferative capacity was consistent with the growth characteristics of MSCs and multilineage differentiation potential was identified.CONCLUSION:A novel method to efficiently isolate and culture hNP-MSCs,PELD combined with explant culture method,was established, which would promote the study of regenerative medicine based on hNP-MSCs.

Objective To investigate the killer cell lg-like receptors (KIR) gene frequency of HIV-1 infected slow progressors(SP) and typical progressors(TP), and to analyze the interaction between KIR alleles and the progression of HIV-1 infection in Chinese population. Methods Eighty-one HIV-1 posi-tive individuals including 43 SPs and 38 TPs were recruited. Carriage of KIR genes was assessed using poly-merase chain reaction sequence-specific primers (PCR-SSP) assays. Results KIR2DS3 gene frequency was significantly lower in SP group (3.6%) than that in TP group (14.2%), P =0. 018 ,OR =0. 210,95% CI =0.053-0.833. The number of activating KIR genes was less in SP group than that in TP group, but was not significant (P = 0. 208). Conclusion Lower KIR2DS3 gene frequency may potentially be associated with slower progression to AIDS in Chinese population.

Objective To study the relationships between neutralizing antibody response against heterologous virus and disease progression in Chinese HIV-1 B'/C infected individuals. Methods Plasmas from HIV-1-infected individuals, grouped as HIV chronically infected or AIDS according to CD4+ count and clinical symptom, were tested for neutralizing activity against the three HIV-1 isolates with very low homology in vitro. Six two-fold dilutions of each plasma sample (from 1/10 to 1/320) were tested against each virus from the panel. Giving a 50% reduction in p24Ag compared with normal human plasma control wells was defined as positive. The breadth of the cross-neutralizing response was defined based on the number of viruses that were effectively neutralized by any given patient-derived plasma sample. The magnitude of the crossneutralizing response was defined based on the average neutralizing titer against all heterologous viruses. Resuits We found that there revealed a significant difference between HIV chronically infected and AIDS group in the breaths and magnitudes of neutralizing heterologous virus. There was higher prevalence for the frequency of neutralizing heterologous virus in HIV chronically infected than AIDS. The results showed that there was positive correlation between the breadths and magnitudes of neutralizing response against heterologous virus and the plasma HIV RNA level in HIV chronically infected group, while not in AIDS group. There was no association between the breadth of the neutralizing responses against heterologous virus and CD4 T cell counts. Conclusion The capacity of neutralizing antibodies against heterologous virus varied among different disease stage. There were higher titers of neutralizing antibodies in HIV chronically infected than AIDS group. The loss of neutralizing antibodies in plasma from AIDS group appears to be associated with a narrowing of the antibody response during disease progression. These suggest that the presence of neutralizing antibodies against hetreologous virus was associated with disease progression.