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OBJECTIVE：To study the effects of Plantago asiatica polysaccharide on the proliferation ，migration and invasion of breast cancer cells ，and to investigate its mechanism preliminarily. METHODS ：Using human breast cancer cell MDA-MB- 231 as subjects ，MTT method was adopted to detect the effects of different concentrations of P. asiatica polysaccharide（8，16，32，64 mg/L）on the cell proliferation ability ，and survival rate of the cells was calculated. Scratch test and Transwell invasion test were used to detect the effects of different concentrations of P. asiatica polysaccharide（8，16 mg/L）on cell migration ability and invasion ability. Western blot assay was used to detect the expression of epithelial-mesenchymal transition （EMT）-related proteins [matrix metalloproteinase- 2（MMP-2），MMP-9，E-cadherin，N-cadherin，vimentin]. RESULTS ：Results of MTT assay showed that survival rate of the cells in 32，64 mg/L P. asiatica polysaccharide groups were significantly lower than control group （P＜0.05 or P＜0.01），so that 8，16 mg/L，which did not affect the cell survival rate ，were used as the follow-up drug concentrations. Compared with control group ，relative mobility （12，24 h），relative invasion rate and relative expression of MMP- 2，MMP-9， N-cadherin and vimentin protein were decreased significantly in 8，16 mg/L P. asiatica polysaccharide groups （P＜0.05 or P＜ 0.01），while relative expression of E-cadherin protein was increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：P. asiatica polysaccharide can inhibit the proliferation of breast cancer cells MDA-MB- 231，and inhibit the migration and invasion of the cells by regulating the expression of metastasis and EMT-related proteins.
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Objective:To investigate the changes in gut microbiota diversity with age in elderly people using high-throughput sequencing.Methods:Ninety healthy volunteers were recruited. People who were <60 years old (middle-aged group) were set up as a baseline control group (Age A group), while those aged ≥60 years old were further divided into four groups (60-<70: Age B group, 70-<80: Age C group, 80-<90: Age D group, ≥90: Age E group). Fecal samples were collected to extract DNA. The second-generation sequencing technology was used to amplify and sequence the V3-V4 hypervariable region of 16S rDNA. Bioinformatics analysis was performed to compare the differences in gut microbiota and functional genes among groups.Results:At the phylum level, gut microbiota were composed mainly of Firmicute, Bacteroidetes, Proteobacteria and Actinobacteria in different groups. The proportion of Firmicute was the highest, accounting for over 60%, followed by that of Bacteroidetes. At the genus level, the abundance of Faecalibacterium genus decreased with age. The α diversity analysis showed that the gut microbiota in the elderly of different ages had higher abundance and uniformity, and there was no significant difference among groups. However, the β diversity analysis showed that in community structure there was difference between Age A and Age B groups, and similarity between Age B and Age C groups. Conclusions:The community structure of gut microbiota changed significantly between young and middle-aged people and the elderly over 60 years old. It tended to be relatively stable in people of 60-80 years old, but changed again when they were over 80 years old. Chronic inflammatory diseases, metabolic diseases and tumors in the elderly might be associated with the decrease in Faecalibacterium.
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Objective To investigate the gut microbiota diversity between the elderly supported by institution-based care and home-based care. Methods Fresh stool samples were collected from 18 aged per-sons supported by institution-based care (G1 group), 20 aged persons with home-based care (G2 group) and 20 middle-aged and young adults (G3 group). The V3-V4 hypervariable region of 16S rDNA was ampli-fied and sequenced by next generation sequencing technology. Operational taxonomic units ( OTUs) were an-alyzed by QIIME analysis platform for species annotation, diversity analysis, and inter-group difference anal-ysis. Statistical analysis was performed using RStudio software. Results The top 6 microbiological taxa in the three groups were Firmicute, Bacteroidetes, Proteobacteria, Actinobacteria, Fusobacteria and Verru-comicrobia. The abundance of the Firmicute in the G1 and G2 groups showed significant differences [(61. 47±5. 58)％ vs (76. 55±3. 64)％, P<0. 05]. The G1 and G3 groups had a statistically significant difference in the abundance of the Proteobacteria [(9. 59±12. 68)％ vs (2. 15±2. 47)％, P<0. 05]. The abundance of both Bacteroidetes and Proteobacteria was higher in the G1 group than in the G2 group without significant difference between the two groups. No significant differences in diversity indices ( Shannon, Simpson and Chao1) were found between G1 and G2 groups (P>0. 05). Results of the NMDS analysis showed that the intra-group differences were greater than inter-group differences in G1 and G2 groups. Con-clusions No significant difference in the diversity of gut microbiota was detected between the elderly sup-ported by institution-based care and home-based care, but there were differences in the composition of the predominant gut microbiota.
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Objective@#To investigate the gut microbiota diversity between the elderly supported by institution-based care and home-based care.@*Methods@#Fresh stool samples were collected from 18 aged persons supported by institution-based care (G1 group), 20 aged persons with home-based care (G2 group) and 20 middle-aged and young adults (G3 group). The V3-V4 hypervariable region of 16S rDNA was amplified and sequenced by next generation sequencing technology. Operational taxonomic units (OTUs) were analyzed by QIIME analysis platform for species annotation, diversity analysis, and inter-group difference analysis. Statistical analysis was performed using RStudio software.@*Results@#The top 6 microbiological taxa in the three groups were Firmicute, Bacteroidetes, Proteobacteria, Actinobacteria, Fusobacteria and Verrucomicrobia. The abundance of the Firmicute in the G1 and G2 groups showed significant differences [(61.47±5.58)% vs (76.55±3.64)%, P<0.05]. The G1 and G3 groups had a statistically significant difference in the abundance of the Proteobacteria [(9.59±12.68)% vs (2.15±2.47)%, P<0.05]. The abundance of both Bacteroidetes and Proteobacteria was higher in the G1 group than in the G2 group without significant difference between the two groups. No significant differences in diversity indices (Shannon, Simpson and Chao1) were found between G1 and G2 groups (P>0.05). Results of the NMDS analysis showed that the intra-group differences were greater than inter-group differences in G1 and G2 groups.@*Conclusions@#No significant difference in the diversity of gut microbiota was detected between the elderly supported by institution-based care and home-based care, but there were differences in the composition of the predominant gut microbiota.
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Objective To translate Moral Distress Scale(MDS-R), and to test the reliability and validity of the Chinese version of MDS-R. Methods The MDS-R was translated, back translation and adapted according to Chinese culture. The reliability and validity of Chinese version of MDS-R was tested in 750 nurses in Quzhou city by item correlation analysis, content validity, exploratory factor analysis, confirmatory factor analysis, Cronbach′s Alpha coefficient and test-retest reliability. Results The internal consistency coefficient of the Chinese version of MDS-R ranged from 0.119-0.756 (P<0.01). The content validity was 0.952. validity.Factor analysis extracted three common factors, which explained 64.537％ of variance of the total scale. Based on the exploratory factor analysis, a theoretical model was established for the scale and each factor, and the fitting degree of the theoretical model was verified by the data. After fitting the model, the fitness values of the first and the second order confirmatory factor analysis were all up to the standard level. The Cronbach's Alpha coefficient of the scale was 0.925, and the test-retest reliability was 0.900. Conclusions The Chinese version of MDS-R is reliable and valid, and can be used to measure the moral distress of nurses.
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Objective:To investigate the change of liver function,viral load and CD4+T count in pediatric AIDS patients with HBV/HCV co-infection after HARRT therapy,and explore the effect of HBV/HCV co-infection on HAART.Methods:95 pediatric AIDS patients without HBV/HCV co-infection ( group A) ,9 pediatric AIDS patients with HBV co-infection ( group B) and 23 pediatric AIDS patients with HCV co-infection ( group C) who received HAART for 2 year were enrolled.Liver function,viral load and CD4+T count were detected before and after HAART.Results:After HAART for 2 years,26 patients (20.5%) were found with liver injury of grade 2 (1000.05 ) .Conclusion: Co-infection of HBV/HCV can aggravate the liver damage of HIV-1 infected children,but has no significant effect on HAART.
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Objective To explore the role of glycogen synthase kinase (GSK)-3β/β-catenin signaling pathway on human immunodeficiency virus 1 (HIV-1) negative factor (Nef) protein promoting of human herpesvirus-8 (HHV-8) viral interleukin-6 (vIL-6)-induced angiogenesis.Methods GSK-3β mutant plasmid GSK-3β-S9A,dominant negative (DN) form GSK-3β-DN and the control vector pcDNA3.1+ were transfected into endothelial cells which stably expressed HHV-8 vIL-6 or HIV-1 Nef,or co-expressed vIL-6 and Nef protein.Microtubule formation assay was performed to explore microtubule formation ability.A chick embryo chorioallantoic membrane (CAM) model was used to detect angiogenesis.The expression of GSK-3β/β-catenin signaling pathway-related kinases in transfected cells and CAM tissue were further detected by Western blot.The measurement data were compared by t test.Results The activity of GSK-3β was decreased and the ability of HIV-1 Nef protein was enhanced by transfection with GSK-3β-DN in promoting vIL-6 induced microtubule formation (3.42 vs 2.51,t =3.67,P＜0.01) and angiogenesis (6.25 vs 3.97,t=4.06,P＜0.01).In contrast,the activity of GSK-3β was significantly increased and these functions of HIV-1 Nef protein mentioned above were inhibited by transfection with GSK-3β-S9A (0.62 vs2.51,t=8.48,P＜0.01; 0.39 vs 3.97,t=8.59,P＜0.01).The results of Western blot showed that with the elevated level of,β-catenin (in cells:3.53 vs 2.07,t=6.60,P＜0.05; in tissues:2.76 vs 1.74,t=17.40,P＜0.01) and vascular endothelial growth factor (VEGF,in cells:2.68 vs 1.87,t=4.28,P＜0.01; in tissues:2.20 vs 1.39,t=7.08,P＜0.01) were increased in the GSK-3β-DN transfected cells or tissues,while the opposite results were achieved in the GSK-3β-S9A-transfected cells (GSK-3β phosphorylation:0.50 vs 1.47,t=7.33,P＜0.01; β-catenin:1.05 vs 2.62,t=29.50,P＜0.01; VEGF:0.74 vs 2.16,t=20.95,P＜0.01) or tissues (GSK-3β phosphorylation:0.35 vs 1.97,t=10.72,P＜0.01; β-catenin:0.79 vs 1.77,t=5.72,P＜0.01; VEGF:0.43 vs 1.65,t=11.89,P＜ 0.01).Conclusion GSK-3β/β-catenin signaling pathway is involved in vIL-6-induced angiogenesis promoted by HIV-1 Nef protein,which would be valuable for the therapy of Kaposi's sarcoma,an acquired immunodeficiency syndrome,as a potential molecular target.
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Objective To investigate whether HIV-1 Nef could promote the angiogenesis and tu-morigenesis induced by KSHV vIL-6 through regulating PTEN/PI3K signaling pathway .Methods Lipo-some transfection was used to transfect cDNA of pPTEN , dominant-negative ( DN) construct of PI3K and control vector into endothelial cells , which stably express KSHV vIL-6 and HIV-1 Nef.Microtubule forma-tion assay and chicken chorioallantoic membrane ( CAM) assay were used to evaluate microtubule formation and angiogenesis , respectively .Expressions of PTEN and PI 3 K were measured by Western blot .Results Both overexpression of PTEN and inhibited expression of PI 3K suppressed the vIL-6-induced microtubule for-mation and angiogenesis in CAM mediated by Nef .Conclusion HIV-1 Nef enhances vIL-6-induced angio-genesis and tumorigenesis through regulating PTEN/PI3K signaling pathway .
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ObjectiveTo investigate whether the cellular immune function of athymia nude mice can be reestablished and whether the recipients' specific immune tolerance can be induced by the combined xenotransplantation of embryonic thymus and metanephros anlagen from rats to athymia nude mice.Methods Male and female rats were mated overnight.The presence of vaginal plaques was marked as the embryonic day 0(E0).The whole embryos from Lewis rats on embryonic day 15(E15) were taken for the present experiment.The Lewis E15 metanephros and thymus anlagen were taken out from the embryos and immediately preserved in normal saline at 4℃,and then were implanted into the omentum and renal capsule of nude mice.10 weeks post-implantation,the peritoneotomy was carried out on some recipients,metanephros and thymus transplants were removed for histopathological examination.The concentrations of urea nitrogen and creatinine in cyst fluid from the metanephros transplants were measured.The survival time of the recipients was observed after nephrectomy.The blood of the recipients was obtained to detect the re-establishment of T lymphocyte with flow cytometry(FCM).The lymphocyte suspension extracted from recipients were mixed respectively with the spleen cells suspension of Lewis rats,BN rats or C57BL/6 mice to perform mixed lymphocyte reaction(MLR) test.The skins of Lewis rats,BN rats,C57BL/6 and nude mice were transplanted in the chest-back of the recipients to observe the reject reaction.Results10 weeks post-implantation,both E15 metanephros and thymus transplants were enlarged,and showed well-developed structure under optical microscope and electronic microscope.Metanephros displayed partial excreting function.The concentrations of urea nitrogen and creatinine in cyst fluid from the metanephros transplants were close to the concentrations in the bladder urine.The combined transplant recipients survived longer than the others after nephrectomy(P ＜0.05).The re-establishment of T lymphocyte from the blood of recipients was certified by FCM.MLR indicated that the lymphocyte of the combined transplant recipients had normal reaction to the spleen cells of the BN rats and C57BL/6 mice,but had under-reaction to the donor Lewis's( P＜0.01 ).Survival time of the skin from donor Lewis is longer than that of the BN rats' and the C 5 7 BL/6 mice' s ( P ＜ 0.01 ).Conclusion E15 Lewis metanephros and thymus anlagen heterografted into the athymia nude mice will grow and function well.The nude mice with combined transplantation of E15 Lewis thymus and metanephros anlagen can reestablish immunologic function and may induce tolerance to the metanephros and skin graft from donor Lewis
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Objective To establish antigen capture ELISA methed to detect Kaposi's sarcoma-as-sociated herpesvirus(KSHV)antigen,and to evaluate its feasibility for clinical application.Methods The BALB/c mice and New Zealand white rabbits were injected with purified recombinant KSHV gpK8.1 proteins to prepare the monoclonal antibody(McAb)and polyclonal antibody(PcAb)anti-gpK8.1,respectively.A new antigen capture ELISA method was established for KSHV antigen detection.The detection reproducibili-ty as well as the sensitivity and specificity of this new assay were determined by the optimization test,which antibody pairs were analyzed to choose the best coating antibody and detecting antibody.The 3 KSHV posi-tive patients sera and 257 patients sera from sexually transmitted disease,cancers or gynecological diseases were detected with this assay to evaluate its value for clinical application.Results When the McAb as coat-ing antibody at concentration of 5 μg/ml and PcAb as detecting antibody at concentration of 1.6μg/ml were selected,the highest P/N value could be obtained.The sensitive analysis of this test could detect recombi-nant KSHV gpK8.1 antigen of 31.28 ng/ml.Meanwhile,it is highly specific to detect KSHV antigen with-out cross reaction to Epstein-Barr vims(EBV),herpes simplex virus(HSV)-1 or HSV-2.All of three KSHV-positive sera and 4 sera from 257 clinical samples were positive with this new assay.which indicated that it could be used for capturing KSHV antigen.Conclusion A sensitive and specific McAb-based anti-gen capture ELISA method to detect KSHV antigen were established successfully.It is of great potential val-ue to develop reagent for KSHV clinical serologic dingnosis.
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OBJECTIVE To explore the genotype and distribution of humam papilloma viruses(HPV) among rural women with cervical lesion.METHODS The cervical exfoliated cell specimens were collected and divided into two groups,the experimental group with 340 rural women finally diagnosed as cervical intra-epithelial neoplasm(CIN) or higher grade pathological changes in healthy examination,and the health control group with 230 rural women randomly selected from the crowd taken healthy examination.DNA was extracted and the genotypes of HPV-DNA were monitored by traditional nested PCR,flow-through hybridization and gene chip technique.RESULTS One-hundred and ninety-five cases(57.4%) in experimental group and the 58 cases(25.2%) in control group were confirmed to be HPV-DNA positive.There was significant difference between the two groups(P