Objective To determine the CD4+CD25+Foxp3+T regulatory (Treg) cell levels in peripheral blood (PB) of patients with autoimmune diseases (AID) and age-and sex-matched healthy controls. Methods Clinical data and laboratory examinations of AID cases (n=1561) and healthy controls (n=196) were enrolled. The levels of PB Treg cells, other T lymphocyte subsets [total T, CD4+T, CD8+T, T helper1 (Th1), T helper 2 (Th2), and T helper17(Th17) cells] and clinical indicators, laboratory test results were analyzed retro-spectively. Data were analyzed by t test, Mann-Whitney U test,χ2 test and Spearman correlation analysis. Results ①The absolute counts of Treg [22.9(18.31, 36.47) vs 30.24(21.85, 41.34), Z=-3.974, P<0.01], total T (Z=-4.234, P<0.01), CD8+T (Z=-3.801, P<0.01), Th17 (Z=-3.740, P<0.01) cells in PB of patients with AID were significantly lower than those of healthy controls, the levels of CD4+T/Treg (Z=-3.366, P=0.001), Th1/Treg (Z=-3.213, P=0.001) and Th2/Treg (Z=-2.490, P=0.013) in PB of AID patients were higher than those of healthy controls.② The levels of inflammatory indicators were associated with numbers of T lymphocyte subsets. ③ The levels of Treg (Z=-3.624, P<0.01), total T (Z=-2.954, P=0.009), CD4+T (Z=-3.005, P=0.003), Th2 (Z=-1.896, P=0.049) cells in PB of the patients who had been treated with hormones and/or biological or non-biological disease-modifying anti-rheumatic drugs (DMARDs) were significantly lower while the levels of total T/Treg (Z=-2.460, P=0.014), CD8+T/Treg (Z=-3.197, P=0.001) in PB were higher than those of the primary treatment patients. ④ The levels of Treg (Z=-7.105, P<0.01), total T (Z=-2.954, P<0.01), CD4+T (Z=-6.909, P<0.01), Th1 (Z=-4.875, P<0.01), Th2 (Z=-5.751, P<0.01), Th17 (Z=-5.121, P<0.01) cells in PB of the patients with important organs involvement were lower while the ratios of total T/Treg (Z=-4.500, P<0.01), CD8+T/Treg (Z=-5.925, P<0.01) were higher than those non-organ involvement patients. ⑤ The absolute counts levels of T lymphocyte subsets in the AID patients were not significantly correlated with whether there was a single AID and/or multiple AID overlaps. Conclusion The absolute number of peripheral Treg cells decreases significantly in AID, and is correlated with inflammatory indicators. Patients with retreated and organ involve-ment have fewer Treg cells. Our results suggest that Treg cells may play an important role in the pathogenesis of AID.

Objective To investigate the distribution of death among human immunodeficiency virus/acquired immuno deficiency syndrome(HIV/AIDS) cases in Guizhou Province from 1995 to 2017. Methods The HIV/AIDS death cases from 1995 to 2017 were downloaded from “Chinese National Comprehensive HIV/AIDS Prevention and care Information system” in Guizhou Province and were analyzed. Results From 1995 to 2017, Guizhou Province reported a total of 43 794 HIV/AIDS cases and 11 527 deaths according to current address. After excluding missing persons, the HIV/AIDS mortality rate of the province was 29.8%. The proportion of reported HIV/AIDS cases died in the same year ( 21995-2012=139.5, P<0.001; 22012-2015=28.2, P<0.001) and the proportion of HIV/AIDS cases ( 21995-2012=109.1, P<0.001; 22012-2014=57.2, P<0.001) who survived at the beginning but died later in the year all showed a trend being low-high-low. In the analysis of the detection history of death cases, the detection proportion of cluster of differentiation 4(CD4) T-cell and the proportion of antiviral treatment had been increasing year by year. The analysis of the cause of death found that the proportion of death caused by AIDS increased firstly and then declined, and the proportion of death due to excessive drug abuse showed a trend of declining year by year. Conclusions The mortality rate of HIV/AIDS in Guizhou Province was still high, and decreased rather slow. Expanding the coverage of HIV monitoring and screening is one of the key tasks of AIDS prevention and control. CD4+T-cell testing and free antiviral treatment should be strengthened to reduce the mortality rate of HIV/AIDS in Guizhou Province in the future.

Objective To find late diagnosis and its influencing factors of newly reported HIV/AIDS in Guizhou Province from 2014 to 2018. Methods Through the Chinese National Comprehensive HIV/AIDS Prevention and Care Information System,all newly reported HIV/AIDS cases from 2014 to 2018 in Guizhou Province were analyzed and related factors of late diagnosis were analyzed using binary Logistic regression model. Results From 2014 to 2018, there were 33 611 newly reported HIV/AIDS cases in Guizhou Province, and the late diagnosis rates of newly reported cases were 35.46%, 34.49%, 38.35%, 39.74% and 38.80% respectively. The analysis showed that the proportion of late diagnosis cases from medical institutions increased year by year ( 2=64.603,P<0.001). By analyzing the late diagnosis rate of cases from different sample sources, medical institutions was significantly higher than that reported by voluntary counseling and testing, positive spouses or sexual partners( 2=276.033,P<0.001). Multivariate analysis showed that gender, marital status, route of transmission, occupation, ethnicity and source of samples were associated with the late diagnosis of newly reported cases (all P<0.05). Conclusions It shows a slow upward trend of late diagnosis rate among HIV/AIDS reported in Guizhou Province from 2014 to 2018.On the one hand, it is of great significance to continue to strengthen the publicity and education of the whole population in Guizhou , in order to improve the awareness of HIV active detection. On the other hand, we should continue to expand HIV testing in Guizhou Province to improve the early detection level of HIV/AIDS.

The study was designed to synthesize a novel dendritic copolymer composed of polyamidoamine dendrimer G0 as the inner core and poly(L-glutamic acid) grafted low molecular weight polyethylenimine (PGLP) as surrounding arms for gene delivery vector. The molecular structure of PGLP was confirmed by ¹H NMR (proton nuclear magnetic resonance spectroscopy). The DNA combination capability of PGLP was examined by gel retardation electrophoresis. The particle sizes and zeta potentials of PGLP/pDNA complexes were determined by dynamic light scattering (DLS). The cytotoxicity of PGLP was evaluated by Cell Counting Kit-8 (CCK-8) and hemolysis assays, which was approved by Research Ethics Committee of the First Affiliated Hospital of Nanchang University. The in vitro transfection efficiency of PGLP was measured by a flow cytometry. The results of physicochemical properties suggested that PGLP could self-assemble with DNA to form complexes with average particle sizes of about 105-200 nm and zeta potentials of about +10 - +28 mV, which could protect DNA from serum degradation. The results of biological properties suggested that PGLP showed more higher transfection efficiencies but lower cytotoxicity than PEI 25K or Lipofectamine 2000 in various cell lines (HEK 293T, HeLa, BEL 7402, RASMC). Importantly, it was found that PGLP/pDNA complexes at w/w = 8 showed more strong serum-resistant capacity than PEI 25K/pDNA complexes. Therefore, PGLP is a promising candidate vector for gene delivery.

Objective To explore the expression and significance of vitamin D (VitD) in patients with rheumatoid arthritis (RA),and analyze the relationship between its expression and clinical indicators.Methods Clin-ical parameters and laboratory examinations of RA cases (n=250) were collected.Clinical parameters included were gender,age,disease course,swollen joints number,tenderness joints number,visual analog pain score (VAS),disease activity score (DAS)28 score.Laboratory examinations included erythrocyte sedimentation rate (ESR),C-reactive protein (CRP),rheumatoid factor (RF) and anti-cyclic citrullinated peptide (CCP) antibody,antinuclear (ANA) antibody,antikeratin (AKA) antibody,anti-perinuclear factor (APF),anti-mutated citrullinated vimentin (MCV),antibody and anti-6-glucose phosphate isomerase (GPI) antibody,lymphocyte subsets in the peripheral blood and lymphocyte subsets of CD4+T cells.The level of 25-(OH)-Vit-D and clinical parameters,laboratory examinations were analyzed retrospectively.One-way ANOVA and KruskalWallis test were used for comparison among the groups;and the correlation analysis was performed by Pearson and Spearman rank correlation analysis.Results ① The level of 25-(OH) D in RA patients was significantly lower than that in healthy controls (t=11.676,P＜0.01).② According to 25-(OH)D level,RA patients were divided into the deficiency group,insufficient group and normal group,the tender joints count (x2=17.793,P＜0.001),the number of swollen joints (x2=12.635,P=0.002),ESR (F=6.330,P=0.002),VAS score (F=5.095,P=0.007,DAS28 (F=4.990,P=0.008) were different significantly amorg the three groups.③RF (x2=6.742,P=0.034) and anti-CCP antibody (x2=6.836,P=0.033) were different significantly among the three groups and the level of 25-(OH) D was negatively correlated with RF (r=-0.202,P=0.001),anti-CCP antibody (r=-0.220,P＜0.01),anti-MCV antibody (r=-0.109,P=0.002) and AKA (r=-0.215,P=0.001).④ The level of 25-(OH) D in the RF (t=-2.715,P=0.007),anti-CCP antibody (t=-2.03,P=0.044),AKA (t=-2.108,P=0.036) negative group was significantly higher than that in patients with antibody positive group.⑤ The level of Th1 (IFN-γ) cells (F=3.259,P=0.043) and Treg (CD4+CD25+Foxp3+) cells (F=4.342,P=0.031) were significantly different among the three groups and the level of 25-(OH) D was positively correlated with Treg (CD4+CD25+Foxp3+) cells (r=0.146,P=0.025).Conclusion Vitamin D is generally deficient in RA patients,which is significantly correlated with disease activity,RF,anti-CCP antibody,anti-MCV antibody,AKA and Th1,Treg cells.It is suggested that vitamin D may play an important role in the immunological pathogenesis and disease progression of RA.

Objective To explore the expression and their significance of peripheral Th17 cells and regulatory T cells (Tregs) in idiopathic inflammatory myopathy,and analyze the relationship between the expression and clinical indicators,imaging and pathological changes.Methods Clinical data,laboratory tests,imaging and pathological changes of IIM cases (n=85) and healthy controls (n=70) were enrolled.Clinical data included the classification,age,gender,course of the disease;laboratory tests including erythrocyte sedimentation rate (ESR),C-reactive protein (CRP),creatine kinase (CK),creatine kinase isoenzyme-MB (CKMB),lactate dehydrogenase (LDH),hydroxybutyrate dehydrogenase (HBDH).The level of peripheral Th17,Treg cells and clinical indicators,laboratory tests,imaging and pathological changes were analyzed retrospectively.Since the data was disregarded from the normal distribution,the median four quantile method was used for statistical description.Two samples were compared with Mann-Whitney U test,and the correlation between variables was Spearman rank correlation analysis.Results ①) The levels of Th17 cells in the case group was not significantly different from that in the control group [6.18(3.42,13.65) cell/μl vs 7.42(5.02,11.13) cell/μl,P＞0.05],the levels of Treg cells in patients was significantly lower than that in the control group [21.25(12.48,35.67) cell/μl vs 36.95(30.37,47.12) cell/μl,P＜0.05],the ratio of Th17/Treg was also significantly higher than that in the control group [0.31(0.21,0.47) vs 0.18(0.14,0.31),P＜0.05].② Peripheral Treg cells levels were not correlated with ESR,CRP,CK-MB,LDH and HBDH (P＞0.05).Peripheral Treg cells levels were negatively correlated with CRP (r=-0.279,P＜0.05),but no correlated with ESR,CK-MB,LDH and HBDH (P＞0.05).③ According to the involvement of important organs,patients were classified into two groups:organ involvement group and non-organ involvement group.The levels of Treg cells in the organ involvement group was fewer than that in non-organ involvement group [16.54(8.84,27.34) cell/ul vs 24.87(14.44,43.37) cell/ul,P＜0.05],and the ratio of Th17/Treg in the organ involvement group was significantly higher than that in non-organ involvement group [0.41(0.29,0.68) vs 0.29(0.19,0.39),P＜0.05].④) Peripheral Th17 cells levels in patients with skeletal muscle inflammatory edema was significantly higher than that of non-inflammatory edema patients [10.70 (4.11,14.51) cell/μl vs 3.10 (1.27,5.15) cell/μl,Z=-2.460,P＜0.05].⑤ The levels of Th17,Treg cells and ratio of Th17/Treg did not correlate with pathological features of inflammatory infiltration (P＞0.05).Conclusion The absolute number of peripheral Treg cells decreases significantly in IIM,and correlates with CRP.Patients with organ involvement have fewer Treg cells,and there is imbalance between Th17 and Treg.When muscle MRI presents with inflammatory edema,patients may have high level of Th17 cells.Our results suggest that Treg cells may play an important role in the pathogenesis of IIM.

Objective Post-transcription RNA interference (RNAi) is more and more widely applied in multigene therapy.This study aimed to establish an subcutaneous xenotransplanted tumor (SXT) model of human HT-29 colon carcinoma in nude mice and investigate the effects of the survivin shRNA-APC double gene co-expression lentiviral vector on the growth of SXT.Methods Thirty-five nude mice were equally divided into five groups, double-gene survivin shRNA, survivin shRNA, APC, empty vector, and blank, and injected into the left anterior axillary with respective stably transfected cell lines and human HT-29 colon carcinoma cells, all at 2×106/mL, to establish an SXT model of human HT-29 colon carcinoma.The inhibition rate of tumor growth was calculated by measuring the size and weight of the SXT, the expressions of survivin mRNA and protein in the tumor tissue detected by real time PCR and immunohistochemistry respectively, and the apoptosis of the HT-29 colon carcinoma cells determined by TUNEL.Results The mean size and weight of the SXT were significantly reduced in the double-gene survivin shRNA-APC, survivin shRNA, and APC groups as compared with the blank and empty vector groups (P<0.05), though increased in the survivin shRNA and APC groups in comparison with the double-gene group (P<0.05).The expressions of survivin mRNA and protein in the tumor tissue were remarkably lower in the double-gene survivin shRNA-APC, survivin shRNA, and APC groups than in the blank and empty vector groups (P<0.05), even lower in the double-gene group than in the survivin shRNA, and APC groups (P<0.05).The apoptosis rate of the HT-29 colon carcinoma cells was markedly up-regulated in the double-gene survivin shRNA-APC ([56.72±3.17]%), survivin shRNA ([33.64±2.03]%), and APC groups ([31.19±1.79]%) as compared with the blank ([9.89±0.31]%) and empty vector groups ([10.06±0.43]%) (P<0.05), even more significantly in the double-gene than in the survivin shRNA and APC groups (P<0.05).Conclusion The survivin shRNA-APC double gene co-expression lentiviral vector can reduce the expression level the survivin gene, promote the apoptosis of colon carcinoma cells, and suppress the growth of the subcutaneous xenotransplanted tumor.

Polyamidoamine (PAMAM) dendrimers as synthetic gene vectors are efficient gene delivery systems. In this study, a kind of α-cyclodextrin-PAMAM conjugates polymer (CyD-G1) was synthesized as a gene delivery vector. Based on ¹H NMR detectation, about 6.4 PAMAM-G1 molecules was grafted onto an α-CD core. Agarose gel electrophoresis revealed that CyD-G1 could efficiently bind with DNA to condense them into nano-scale particles, which showed a similar binding capacity of PEI-25K. Besides, it could protect DNA from DNase I degradation in a low N/P ratio. When N/P ratio in the CyD-G1/DNA polyplex was 40, the average particle size of CyD-G1/DNA polyplex was about 120 nm, and zeta potential was +21 mV. This polyplex could maintain its particle size in serum-containing solution within 360 min. In comparison with PEI-25K carrier, CyD-G1 showed low cytotoxicity in various cell lines. Cell transfection results showed that CyD-G1 efficiently delivered DNA into cells at N/P=80 compared with Lipofectamine 2000 and PEI-25K.Unlike Lipofectamine 2000 and PEI-25K, in serum-containing test condition, CyD-G1/DNA polyplex could maintain the transgene activities. The results of confocal laser scanning microscopy indicated that most DNA entered into cell nuclei within 4 h, and this phenomenon was consistent with the results calculated by flow cytometry. Taken together, CyD-G1 showed good transgene activities and the gene delivery vector could be used not only in vitro but also in vivo.