Objective:HIV-1 infection was associated with a variety of host genetic factors ,and HLA was one of the factors that contribute to the progression of disease .We analyze the frequency of HLA-A, HLA-B, HLA-C in MSM cohort , and the relationship between HLA gene with disease progression .Methods:PCR-sequence specific primer typing HLA typing was used to detect the allele frequency of HLA gene in 310 patients.HIV Molecular Immunology Database ( HLA Analysis Tools ) to analysis the frequency of HLA.Results:In MSM cohort,HLA-A*1101(34.52%) gene was the highest,followed by HLA-A*0201(31.94%),HLA-C*0102 (27.10%) and HLA-A*2402(26.45%).According to the CD4 cell count of HIV infected patients ,the results showed that there were low levels of HLA-B*4403(1.12%,P=0.0276),HLA-B*1511(2.25%,P=0.0282) and HLA-B*5701(0.37%,P=0.0491) in rapid progressors,while the HLA-C*0304(8.96%,P=0.0319) was higher than that of nonprogressors (4.56%).Conclusion: We obtained the distribution frequency data of HLA-A,HLA-B and HLA-C in MSM cohort.Our data presented here may offer potential cor-relation between dominant effect of HLA alleles and HIV-1 disease progression.And found that the HLA-B*4403,HLA-B*1511, HLA-B*5701 related to slow HIV disease progression and HLA-C*0304 related to accelerate HIV disease progression .

Objective To identify the Banna viruses isolated in northwestern part of Yunnan prov-ince in order to make the difference clear between the isolates and other Banna viruses isolated in other parts of Yunnan. Methods Three isolates of Banna vires isolated in 2005 and 2006 were identified by morpholo-gy, RNA-PAGE profile and molecular biologic method. Nueleotide and amino acid sequences of segment 12 of the 3 isolates were sequenced and analyzed. Results Three Banna viruses were isolated from mosquitoes collected in northwestern part of Yunnan during 2005 and 2006. Electron microscopy study showed that they are spherical with a diameter of 70 nm, no envelope but two layers of eapsid. It was found that the genome of the 3 isolates composes of 12 segments presenting band profile of 6-6 in RNA-PAGE. Nueleotide acid se-quence analysis about segment 12 showed that the identity was 99% between the 3 new isolates, 98% and 90% between the 3 isolates and the strains isolated in other parts of Yunnan, China and Indonesia, respec-tively. Phylogenetie analysis based on segment 12 gene showed that 3 new isolates clnstered in the same branch with the viruses isolated in other parts of Yunnan. The same difference of amino acids was found between Banna viruses isolated in China and Indonesia strains in the analysis of segment 12. Conclusion Banna virus strains were firstly isolated from mosquitoes collected in northwestern part of Yunnan province. Nueleotide acid sequence analysis of the 3 new isolates showed higher identity with strains isolated in other parts of Yunnan.

Objective To compare the effect of silver-coated dressing and general aseptic dressing on the wound healing of abdominal operation. Methods A total of 94 patients from the general surgery department of Second Affiliated Hospital of Xi'an Jiaotong University who underwent abdominal operation were divided into two groups by double blind method, 49 cases in silver-coated dressing group, and 45 cases in general aseptic dressing group. The wound healing of the two groups were observed after wound dressing change, and the wound dressing change times, the healing time, the degree of pain, the positive rate of bacterial culture and the incidence rate of scar healing were observed in the two groups. Results The patients in silver-coated dressing and general aseptic dressing group had no significant differences in gender, age, length of incision, type of incision and poor wound healing type (all P>0.05);after two wound dressings change treatment, the wound dressing change times and incision healing timeof the silver-coated dressing group was (6.52 ± 1.52) times and (16.34 ± 5.96) days, which were less than those of the general aseptic dressing group, which were (8.74 ± 2.35) times and (23.32 ± 8.32) days; and the pain severity (NRS) scored 4.13 ± 1.01, which was lower than that of general aseptic dressing group (6.54 ± 0.95), with significant difference between two groups (t=5.482, 4.704, 11.890, P < 0.05). The bacterial culture positive rates of the silver-coated dressing group was 60.00% (21/35), which was higher than that of the general aseptic dressing group, 23.33%(7/30), the difference was statistically significant (P<0.01);at the same time, the scar healing cases was 4 in the silver-coated dressing group, which was less than 11 cases in the general aseptic dressing group. The difference was statistically significant (P < 0.05). Conclusions The silver-coated wound dressing method is better than the general aseptic wound dressing, and has obvious advantages in the treatment of abdominal poor healing incision. The silver-coated wound dressing can promote the healing of incision, reduce scar healing, shorten the hospitalization time, reduce the suffering of patients.

Objective　To construct the eukaryotic expression recombinant plasmid, pcIFN-γ, as a genetic adjuvant and observe the immune responses elicited by pcDNA3-rhoptry protein 1 （pc-ROP1） combined with pcIFN-γ against Toxoplasma gondii (T. gondii) infection in mice. Methods　A fragment of the IFN-γ gene was directly inserted into the pcDNA3 plasmid and identified by two restriction endonucleases digestion. pcIFN and pcROP1 DNA was injected into the left leg muscle of mice at a dosage of 100*!μg, and a booster vaccination was given at the same dosage after two weeks. Control groups were injected with pcDNA3 blank plasmid or normal saline. At 30, 50 and 70 days after booster injection, kinetic tests were carried out: MTT assay for the proliferation response of T lymphocyte cells and the activity of NK cells, sandwich ABC-ELISA for the determination of IFN-γ, IL-2 and IL-10; a serum enzymetic aassay for nitric oxide (NO) in sera and ELISA for the titer of IgG antibody in sera. Results　The recombinant plasmid, pcIFN-γ was constructed. The proliferation response of spleen T lymph cells, NK cell killing activity, and serum levels of IFN-γ, IL-2 and NO in mice injected with pcROP1 and pcIFN-γ were higher than in those injected with pcROP1 alone. There was no difference in IgG antibody levels between the two groups. Conclusion　The genetic adjuvant, pcIFN-γ, could enhance the cellular immune response induced by DNA vaccine of pcROP1 in mice against Toxoplasma gondii infection.

Objective To analyze the differentiation status of CTL and to evaluate its clinical val-ue in patients with HIV/HCV coinfection .Methods Twenty-eight patients with HIV/HCV coinfection and twelve patients with single HCV infection were enrolled in this study .The technique of Fibro-Scan was used to evaluate liver fibrosis .The viral load of HCV was detected by real-time quantitative PCR .Flow cytometry analysis was performed to measure the differentiation status of CTL .Results Both of the levels of alanine transaminase ( ALT) and alkaline phosphatase ( ALP) in patients with HIV/HCV coinfection were signifi-cantly higher than those in patients with single HCV infection [(53.7464±48.1180) U/L vs (27.4750± 13.9850) U/L, P=0.012;(24.5071±8.1940) g/L vs (16.9667±7.1890) g/L, P=0.009].The liver stiffness of patients with HIV/HCV coinfection was higher than that of patients with single HIV infection [(5.9500, 5.8250) Kpa vs (5.1500, 1.0500) Kpa, P=0.117].Compared with the patients with single HCV infection, the patients with HIV/HCV coinfection showed higher viral loads of HCV [( 6.4768, 5.3434) lg copy/ml vs (2.6815, 1.6990) lg copy/ml, P=0.012], but lower clearance rate of HCV [32.14%vs 75%, P=0.032].Compared with the patients with single HCV infection , the patients with HIV/HCV coinfection showed lower percentages of CD 27+CD28+CTL [(28.265±15.095)%vs (18.068±10.263)%, P=0.017), but higher percentages of CD27+/-CD28+CTL [(62.449 ±14.561)% vs (71.111±12.681)%, P=0.066].A trend toward negative correlation was observed between the percent -age of CD27+CD28+CTL and the degree of liver stiffness (r=-0.310, P=0.058).Conclusion HIV in-fection could accelerate the progression of liver disease in patients with HIV /HCV coinfection by affecting the differentiation of CTL .

OBJECTIVETo characterize the immune response induced by SAG1 encoding plasmid combined with IL-2 gene adjuvant in mice and to assess the protective effect of this vaccination against toxoplasmosis.

METHODSMice were co-injected intramuscularly with plasmid encoding Toxoplasma gondii SAG1 plus murine IL-2 expression vector at a dose of 100 microg. Booster immunizations were employed 2 more times at 3-week interval. As controls, mice were inoculated with PBS or empty plasmid pcDNA3. Humoral and cellular responses were assayed using ELISA for the determination of Ab, Ab isotype and IFN-gamma, as well as IL-4. To detect the integration and dissemination of DNA in the injected mice, PCR and in situ hybridization were performed. All mice were then infected with highly virulent RH tachyzoites of Toxoplasma gondii intraperitoneally.

RESULTSSignificant increases in specific IgG levels were observed in mice after immunization three times with SAG1 expression plasmid. With respect to the IgG isotype, co-inoculation of IL-2 expression plasmid enhanced the level of IgG2a and the production of IFN-gamma. Challenging mice by vaccinating with combined plasmids with RH tachyzoites resulted in prolonged survival.

CONCLUSIONHumoral and cytokine responses elicited by SAG1 DNA immunization can be modulated by co-inoculation with IL-2 expression plasmid. The use of DNA vaccine in combination with an appropriate cytokine gene to prevent T. gondii infection warrants further investigation.

Animals ; Antibodies, Protozoan ; blood ; Antigens, Protozoan ; Cytokines ; biosynthesis ; Female ; Immunization ; Immunoglobulin G ; blood ; classification ; Interleukin-2 ; genetics ; Mice ; Protozoan Proteins ; genetics ; Protozoan Vaccines ; immunology ; Toxoplasma ; immunology ; Toxoplasmosis, Animal ; prevention & control ; Vaccines, DNA ; immunology

Objective To understand the influence of HIV infection on hepatitis C progress in patients co-infected with HIV and hepatitis C virus (HCV) and related immune mechanism. Methods Twenty eight patients co-infected with HIV/HCV and 12 patients with simplex HCV infection were enrolled. The liver function and hepatic fibrosis progress were evaluated by detecting peripheral blood and with Fibro-Scan. The viral load of HCV was detected by using real time quantitative PCR. And the percentage of Treg/CD4 +T lymphocyte cell was tested by using flow cytometry. Results The levels of ALT and ALP in HIV/HCV co-infection group were (76.16 ± 81.248)U/L,(24.507 1 ± 8.194)g/L respectively,higher than those of simplex HCV infection group [(27.475 0±13.985)U/L,(16.966 7±7.189)g/L],the differences were statistical significant. P value was 0.012 and 0.009 respectively. The liver fibrosis index in HIV/HCV co-infection group was 5.950 0-5.825 0 Kpa,higher than that in simplex HIV infection group(5.150 0-1.050 0 Kpa),and the difference was nearly statistical significant(P=0.077). The HCV viral load in HIV/HCV co-infection group was(6.476 8-5.343 4)lg copy/ml,higher than that in simplex HCV infection group[(1.699 0-2.681 5)lg copy/ml],and the rate of HCV clearance in HIV/HCV co-infection group was 32.14%, lower than that in simplex HCV infection group(75.00%). P value was 0.012 and 0.032 respectively. The percentage of Treg/CD4+T lymphocyte cell in HIV/HCV co-infection group was (7.460 0%-2.287 5%),higher than that in simplex HCV infection group (5.965 0%-2.105 0%),and the difference was significant (P=0.032). The percentage of Treg/CD4 + T lymphocyte cell was significantly related with HCV viral load(ρ=0.350,P=0.027),and HCV viral load was significantly related with the liver fibrosis index(ρ=0.487,P=0.001). Conclusion HIV infection could accelerate the progress of hepatitis C,and Treg cells were involved in this progress.

Objective To investigate the effect of time interval between diagnosis and surgical treatment on the prognosis of breast cancer.Methods A retrospective study that include a total of 252 female patients who underwent breast cancer diagnosis and treatment in the Second Affiliated Hospital of Xi'an Jiaotong University from April 2012 to August 2014 were included in the present study,the average age was (58.2 ± 10.8) years old,range from 31 to 67 years old.General demographic information and data of tumor were collected.Information on postoperative recurrence,metastasis,death,and disease-free survival status of breast cancer patients were followed up 5 years by outpatient follow-up or telephone follow-up.All participants were divided into four groups (＜2 weeks,2-4 weeks,4-8 weeks,≥8 weeks) by the time interval between diagnosis and surgical treatment,including 26,118,78 and 30 cases,respectively.In addition,according to the diameter of breast cancer tumors,all participants were divided into three groups (＜20 mm,20-40 mm,and ≥40 mm),including 99,124,and 29 cases,respectively.According to the results of pathological examination of the lymph nodes obtained during intraoperative dissection,the all participants were divided into three groups (lymph nodes without metastasis,1 to 3 metastasis,and ≥3 metastasis),including 66,124,and 62 cases,respectively.The Cox proportional regression risk models were used to assess the hazard ratio (HR) and its 95％ confidence interval (CI) of time interval between diagnosis and surgical treatment with the prognosis of breast cancer,with adjustment for age,education levels and body mass index.Further,stratified analysis by tumor characteristics,including pathological type,histological grade,tumor diameter,lymph node metastasis,and receptor expression were also conducted to evaluated the above association.Kaplan-Meier survival curve was used to evaluate the effects of time interval between diagnosis and surgical treatment on the prognosis of breast cancer.Results The Kaplan-Meier survival curves for the five-year follow-up of total survival time between 4 different time intervals groups showed significantly different (P ＜0.001),and patients with a pre-treatment interval of ＜2 weeks had the longest survival time,while those with ≥8 weeks had the lowest survival time.With a one-week interval before treatment,the overall risk of death in breast cancer patients increased by 6％ (HR =1.06,95％ CI:1.01-1.1 l),and the risk of breast cancer death increased by 8％ (HR =1.08,95％ CI:1.02-1.14),the risk of distant metastasis of breast cancer cells increased by 10％ (HR =1.10,95％ CI:1.08-1.13).With the increase in breast cancer tumor diameter (＜20 mm,20-40 mm,≥40 mm),the overall risk of death due to prolonged treatment interval increased gradually,with HR (95％CI) were 1.06 (1.03-1.09),1.08 (1.02-1.12) and 1.11 (1.05-1.17),respectively.With the increase of lymph node metastasis in breast cancer (no metastasis,metastasis at 1-3,≥ 3 metastasis),the total mortality risk caused by prolonged treatment time interval also showed an increasing trend,with HR (95％CI) were 1.04 (1.02-1.08),1.06 (1.04-1.08) and 1.08 (0.99-1.11),respectively.The same results were also shown in the effect of tumor diameter or distant lymph node metastasis on the association between treatment time interval and breast cancer survival and distant metastasis of breast cancer cells.Conclusion With the prolongation of the time interval between the diagnosis of the breast cancer and the surgical treatment of breast cancer patients,the risk of postoperative death is significantly increased,and the association is more pronounced in breast cancer patients with larger tumor volume or higher distant lymph node metastasis.

Objective To establish a quantitative assay of serum Golgi protein 73 ( GP73) using xMAP technology and evaluate its performance. Methods Monoclonal antibodies against GP73 were prepared and purified, and antibody pair screening was performed by double?antibody sandwich enzyme?linked immunosorbent assay. The screened antibodies were used to construct a Luminex liquid chip detection system, and the analysis performance of the detection system was evaluated. The serum levels of GP73 were detected in 90 clinical samples from healthy controls and patients with chronic hepatitis B infection ( CHB) and hepatocellular carcinoma ( HCC). Results Five anti?GP73 monoclonal antibodies were prepared and purified, and 5 antibody pairs were successfully screened.The Luminex liquid chip detection system of GP73 was successfully constructed using 8F10D1 and 10B9F11 antibody pairs. The analytical performance evaluation showed that the sensitivity of this system was 0.25 ng／ml and the dynamic range was 0.25?100 ng／ml. No cross reactivity was observed. The intra? and inter?assay variation for GP73 was ＜8% and ＜11%, respectively. The recovery was 83%?92%. The linear regression equation was y＝1.141x+6.436 ( r2 ＝0.998 4, P＜0.001). The GP73 concentrations in the serum samples of healthy control, CHB group, and HCC group were 42.8 (38.68, 55.90) ng／ml, 61.49 (43.59, 81) ng／ml, and 122.78 (49.36 liter, 264.55) ng／ml, respectively.The levels of GP73 in HCC group were significantly higher than those in CHB group and healthy controls (P＜0.05). Moreover, the levels of GP73 in CHB group were significantly higher than those in healthy controls ( P＜0.05). Conclusions A liquid chip detection system of GP73 was successfully constructed. It provides a powerful tool for the clinical application of GP73 in the future.

Objective To establish a quantitative assay of serum Golgi protein 73 ( GP73) using xMAP technology and evaluate its performance. Methods Monoclonal antibodies against GP73 were prepared and purified, and antibody pair screening was performed by double?antibody sandwich enzyme?linked immunosorbent assay. The screened antibodies were used to construct a Luminex liquid chip detection system, and the analysis performance of the detection system was evaluated. The serum levels of GP73 were detected in 90 clinical samples from healthy controls and patients with chronic hepatitis B infection ( CHB) and hepatocellular carcinoma ( HCC). Results Five anti?GP73 monoclonal antibodies were prepared and purified, and 5 antibody pairs were successfully screened.The Luminex liquid chip detection system of GP73 was successfully constructed using 8F10D1 and 10B9F11 antibody pairs. The analytical performance evaluation showed that the sensitivity of this system was 0.25 ng／ml and the dynamic range was 0.25?100 ng／ml. No cross reactivity was observed. The intra? and inter?assay variation for GP73 was ＜8% and ＜11%, respectively. The recovery was 83%?92%. The linear regression equation was y＝1.141x+6.436 ( r2 ＝0.998 4, P＜0.001). The GP73 concentrations in the serum samples of healthy control, CHB group, and HCC group were 42.8 (38.68, 55.90) ng／ml, 61.49 (43.59, 81) ng／ml, and 122.78 (49.36 liter, 264.55) ng／ml, respectively.The levels of GP73 in HCC group were significantly higher than those in CHB group and healthy controls (P＜0.05). Moreover, the levels of GP73 in CHB group were significantly higher than those in healthy controls ( P＜0.05). Conclusions A liquid chip detection system of GP73 was successfully constructed. It provides a powerful tool for the clinical application of GP73 in the future.