HIV self-testing and related factors in men who have sex with men in Shijiazhuang / 中华流行病学杂志

Objective: To understand HIV self-testing and related factors in men who have sex with men (MSM) in Shijiazhuang. Methods: From August to September 2020, convenient sampling was used to recruit MSM in Shijiazhuang. Online questionnaires were used to collect information about their demographic characteristics, sexual behaviors and HIV self-testing. logistic regression model was used to analyze the related factors associated with HIV self-testing. Results: In the 304 MSM respondents, 52.3% (159/304) had HIV self-testing in the past 6 months, and 95.0% (151/159) used fingertip blood HIV detection reagent. Self-purchase was the main way to obtain HIV testing reagents (45.9%, 73/159), followed by supply from MSM social organization (44.7%, 71/159). The reasons for having HIV self-testing were non-specific testing time (67.9%, 108/159) and privacy protection (62.9%,100/159), the reasons for having no HIV self-testing included inability of using (32.4%, 47/145), being unaware of HIV self-testing reagent (24.1%, 35/145), and worry about inaccurate self-testing results (19.3%, 28/145). Multivariate logistic regression analysis showed that being 18-29 years old (aOR=2.68, 95%CI: 1.20-5.94), obtaining free HIV self-testing kits in recent 6 months (aOR=8.61, 95%CI: 4.09-18.11) and making friends through Internet and social software (aOR=2.68, 95%CI: 1.48-4.88) were positive factors for having HIV self-testing. Conclusion: HIV self-testing is a more flexible and convenient way to detect HIV in MSM, and the promotion of HIV self-testing in MSM should be strengthened to further increase the HIV detection rate in this population.

Evaluation of clinical utility of a novel method for quantitative detection of hepatitis B virus RNA / 中华传染病杂志

Objective:To evaluate the clinical utility of a novel quantitative assay named hepatitis B virus (HBV) simultaneous amplification and testing (SAT) using a kit for HBV RNA detection (RNA probes).Methods:HBV RNA was detected in 170 serum samples of chronic hepatitis B patients and 30 serum samples of patients without HBV infection collected by simple random sampling method from June 2017 to June 2018 in Huashan Hospital, Fudan University, Shanghai using HBV SAT and reverse transcription polymerase chain reaction (PCR) method. The sensitivity, specificity, kappa value and quantitative correlation of the two methods were analyzed and compared.The detection rates of HBV RNA from samples with different HBV DNA concentrations of the two methods were analyzed and compared. Statistical analysis was performed by chi-square test.Results:Based on the clinical diagnosis, the detection sensitivity, specificity, kappa value of HBV SAT were 97.06%(165/170), 100.00%(30/30) and 0.908, respectively, while the reverse transcription PCR were 92.94%(158/170), 100.00%(30/30) and 0.798, respectively. Among samples with lower concentration of HBV DNA (HBV DNA<100 IU/mL), the detection rates of HBV SAT and reverse transcription PCR were 77.27%(17/22) and 59.09%(13/22), respectively. The linear correlation coefficient of the two methods was r=0.987 8. Conclusions:Quantitation results of HBV RNA by HBV SAT and reverse transcription PCR method are consistent. HBV SAT is a rapid and accurate method for HBV RNA quantitative detection, which has a slightly higher detection rate than reverse transcription PCR in samples with low concentration of HBV DNA.

Analysis of the changes and efficacy of antiviral treatment regimens in patients with chronic hepatitis C from 2014 to 2019 / 中华传染病杂志

Objective:To analyze the changes and efficacy of antiviral treatment regimens in patients with chronic hepatitis C.Methods:This was a single center retrospective study. A total of 157 patients with chronic hepatitis C in Huashan Hospital, Fudan University from January 2014 to February 2019 were included. Clinical informations of antiviral treatment and follow-up were collected. The sustained virologic response (SVR) rate and adverse events in patients receiving different antiviral regimens were compared. Chi-square test was used for statistical analysis.Results:Among the 157 patients, 133 patients had sufficient follow-up data. Seventy-one patients received treatment before 2017, among which 63 patients received interferon regimens and the SVR rate was 74.65%(53/71). Sixty-two patients received treatment after 2017, among which 61 patients received direct-acting antiviral agents (DAA) regimens and the SVR rate was 98.39%(61/62). The difference in SVR rate between the two groups was statistically significant ( χ2=15.230, P<0.01). In 69 patients who received DAA regimens from 2014 to 2019, the SVR at post-treatment week 12 (SVR12) was 95.65%(66/69). Among 43 patients who received DAA regimens containing sofosbuvir, the SVR12 rates of patients with hepatitis C virus genotype 1, 3 and other genotypes were 15/15, 5/6 and 90.91%(20/22), respectively. All the 26 patients who received DAA regimens non-containing sofosbuvir achieved SVR12. The SVR12 rates of patients with different hepatitis C virus genotypes and DAA regimens were not significantly different ( χ2=5.243, P=0.263). The incidences of adverse events in pre-2017 group and post-2017 group were 84.62%(77/91) and 6.06% (4/66), respectively. The difference was statistically significant ( χ2=94.520, P<0.01). The most common adverse events were decreases in neutrophil cell count, decreases in hemoglobin level and decreases in platelet count. Treatment was ceased in six patients due to adverse events. Conclusions:After 2017, the majority of patients with chronic hepatitis C received DAA regimens instead of interferon regimens. The SVR rate increases and the incidence of adverse events decreases along with the changes of leading treatment regimens.The SVR12 rate is higher in patients receiving DAA regimens, regardless of hepatitis C virus genotypes.

Clinical features of 42 cases of COVID-19 / 上海预防医学

Objective To investigate the clinical features, laboratory findings, chest CT findings and treatment of patients with COVID-19, and to analyze their relationship with prognosis. Methods From January to February 2020, the clinical data on the 42 patients with COVID-19 admitted to the Wenzhou Sixth People′s Hospital were analyzed retrospectively. Results The clinical symptoms of the 42 cases included fever (35 cases), cough (26 cases), fatigue (14 cases), aspiration (9 cases), sore throat (4 cases), muscle ache (5 cases), headache (2 cases), nausea (4 cases), diarrhea (6 cases) and abdominal pain (1 case).The absolute number of blood lymphocyte decreased to different degrees in 22 cases.Fourteen cases had lactate dehydrogenase obviously, with no obvious change in procalcitonin.The imaging manifestations were cloud-like and ground-glass-like high density shadows scattered outside the lungs, small flaky consolidation and bronchus inflating sign were seen locally.A few images showed diffuse high density, most of the lesions showed consolidation or striate change, and local fibrosis was formed in the lower lobes of both lungs. Conclusion Fever and cough are the first symptoms of COVID-19, and a few cases are associated with shortness of breath and diarrhea, accompanied by different degrees of systemic symptoms, but most of the patients improve their conditions after active antivirus, anti-infection, systematic symptoms improvement and supportive treatment.The disease is highly infectious and its condition changes rapidly.Therefore, early detection, early diagnosis and comprehensive treatment of the whole body as soon as possible are the keys to treatment.

HIV/AIDS related mortality in southern Shanxi province and its risk factors / 中华流行病学杂志

<p><b>OBJECTIVE</b>To explore factors influencing mortality rate of HIV/AIDS and to improve the effectiveness of antiretroviral therapy (ART).</p><p><b>METHODS</b>By means of retrospective cohort study and the AIDS control information system, HIV/AIDS case reports and antiviral treatment information of 4 cities in southern Shanxi province up to end of December 2012 were selected, to calculate the mortality rate and treatment coverage based on further data collected, along with analysis using the Cox proportional hazards survival regression.</p><p><b>RESULTS</b>4 040 cases confirmed of HIV/AIDS were included in this study. The average age was (36.0 ± 12.9) years, with 65.3% being male, 56.5% being married, 73.5% having junior high school education or lower, 58.4% being peasants, 54.3% with sexually transmitted infection (40.1% were heterosexual, 14.2% were homosexual), and 38.9% were infected via blood transmission (20.2% were former plasma donors, 16.2% blood transfusion or products recipients, 2.4% were injection drug users). Overall mortality decreased from 40.2 per 100 person/year in 2004 to 6.3 per 100 person/year in 2012, with treatment coverage concomitantly increasing from almost 14.8% to 63.4%. Cox proportional hazards survival regression was used on 4 040 qualified cases, demonstrating the top mortality risk factor was without antiretroviral therapy (RR = 14.9, 95% CI: 12.7-17.4). Cox proportional hazards survival regression was made on 1 938 cases of antiviral treatment, demonstrating that the mortality risk of underweight or obese before treatment was higher than those of normal and overweight cases (RR = 2.7, 95% CI: 1.6-4.5), and the mortality of those having a CD4(+) T-lymphocyte count ≤ 50 cells per µl before treatment was more than 50 cases (RR = 2.6, 95% CI: 1.5-4.5); Cox proportional hazards survival regression was made on 2 102 cases of untreated cases, demonstrating the mortality risk of those initially diagnosed as AIDS was higher than those initially diagnosed as HIV (RR = 3.4, 95% CI: 2.9-4.0).</p><p><b>CONCLUSION</b>The ART could successfully make lower HIV/AIDS mortality rate, indicating effective ART can further decrease mortality.</p>

Advance in modern studies on compatibility of Coptidis Rhizoma and Evodiae Fructus / 中国中药杂志

In traditional clinical application, Coptidis Rhizome and Evodiae Fructus have been combined to treat various stomach heat and cold syndromes, gastritis, gastric ulcer and the like. With the application of modem instruments and the development of molecular pharmacologic theory, their chemical constituents and pharmacological effects have been sufficiently studied. In this paper, literatures from Pubmed were adopted, with particular emphasis on findings of international counterparts and studies on compatibility of main chemical components in Coptidis Rhizoma and Evodiae Fructus, in order to elaborate on the scientific comparability of Coptidis Rhizoma and Evodiae Fructus through chemical analysis, and pharmacological and biopharmaceutics studies and introduce the future development trend of the studies.

Clinical observation of recurrent oral ulcer of heat accumulation in the heart and spleen treated with acupoint injection and fire needling therapy / 中国针灸

<p><b>OBJECTIVE</b>To compare the clinical efficacy between the combined acupuncture therapy of acupoint injection and fire needling and the oral administration of vitamin C and B2 as well as subcutaneous injection of transfer factor.</p><p><b>METHODS</b>One hundred and fifty-eight patients of recurrent oral ulcer (ROU) differentiated as heat accumulation in the heart and spleen were randomized into an observation group and a control group, 79 cases in each, one. In the observation group, Xinshu (BL 15), Pishu (BL 20) and Zusanli (ST 36) were selected for acupoint injection of shuang huanglian injection, 1 mL at each point, three times a week, for 2 weeks totally. Additionally, fire needling was applied to the ulcer lesion and peripheral mucosa, once a day, totally for 3 times. In the control group, vitamin C 100 mg and vitamin B2 20 mg were prescribed for oral administration, three times a day; and subcutaneous injection of transfer factor was applied, 2 mL (3 mg), twice a week, for 2 weeks totally. The score of symptoms and physical signs and the visual analogue scale (VAS) were observed before and after treatment, as well as the time of ulcer healing and ulcer recurrence in the patients of the two groups. The clinical efficacy was compared between the two groups.</p><p><b>RESULTS</b>The total effective rate was 92.4% (73/79) in the observation group, better than 77.2% (61/79) in the control group (P < 0.01). The differences in the score of symptoms and physical signs and VAS score before and after treatment were significant statistically in the observation group and the control group (all P < 0.01). The score of symptoms and physical signs and VAS score after treatment in the observation group were reduced significantly as compared with those in the control group (both P < 0.01). The time of ulcer healing in the observation group was earlier than that in the control group [(3.87 +/- 1.01) d vs. (6.22 +/- 0.53) d, P < 0.01]. The recurrence rate in the observation was lower than that in the control group [22.7% (17/75) vs. 50.0% (36/72), P < 0.01].</p><p><b>CONCLUSION</b>The combined therapy of acupoint injection and fire needling achieves the better efficacy on ROU compared with oral administration of vitamin and subcutaneous injection of transfer factor. This therapy has the good and quick effect of analgesia, relieves the relevant symptoms, shortens the time of ulcer healing and reduces the recurrence rate.</p>

Epigenetic regulatory effects of the methylation of CpG island shore on the expression of lag3 gene in various human T cell lines / 中华微生物学和免疫学杂志

Objective To investigate the effects of DNA methylation on the expression of lympho-cyte activation gene 3 (lag3) in different human T cell lines.Methods A quantitative PCR and a flow cy-tometry analysis were performed to measure the expression of lag3 gene in various T cell lines at mRNA and protein levels.The distribution of CpG sites within the promoter and body of lag3 gene was detected to locate the potential regulatory region(s) (CpG island and CpG island shore).The levels of DNA methylation in each cell line were analyzed.The T cell lines were demethylated with 5-Aza-2′-deoxycytidine (5-Aza-2′-dc) for further investigation on the changes of lag3 gene expression and DNA methylation.Results Jurkat E6-1 cells showed the highest expression level of lag3 gene as compared with J.CaM1.6 and CEM cells.Hyperm-ethylated CpG islands were detected in cells of each cell line.The methylation levels of CpG island shore in J.CaM1.6 and CEM cells were higher than that in Jurkat E6-1 cells.Treatment of J.CaM1.6 and CEM cells with 5-Aza-2′-dc significantly promoted the expression of lag3 gene at mRNA and protein levels as well as the demethylation of CpG island shore.No significant differences with the expression of lag3 gene and the methylation of CpG island were observed in Jurkat E6-1 cells with or without 5-Aza-2′-dc stimulation.Con-clusion Methylation and demethylation of CpG island shore played important roles in regulating the tran-scription of lag3 gene.

Relationship of carotid atherosclerosis with serum brain-derived neurotrophic factor and vascular endothelial growth factor in type 2 diabetes mellitus patients / 第二军医大学学报

Objective To investigate the relationship between carotid atherosclerosis and serum brain-derived neurotrophic factor (BDNF), vascular endothelial growth factor(VEGF)levels in patients with type 2 diabetes mellitus(T2DM). Methods A total of 120 T2DM patients who were treated in our hospital were assigned to non-carotid atherosclerosis group (n=56) and the carotid atherosclerosis group (n=64) according to the results of carotid artery ultrasound detection. Another 60 healthy controls were recruited from health check-up populations. Clinical data of the participants were collected, and their serum levels of BDNF, VEGF were determined by ELISA. Results The serum BDNF levels of health check-up populations, non-carotid atherosclerosis group, and carotid atherosclerosis group were (9.97±2.59) ng/mL, (6.88±3.10) ng/mL,and (5.62±3.41)ng/mL, respectively, showing significant difference between different groups (P<0.05); the serum VEGF levels were (49.29±20.23) ng/mL, (61.29±22.05) ng/mL, and(71.34±20.30)pg/mL, respectively, also showing significant difference between different groups (P<0.05). Conclusion T2DM patients complicated with carotid atherosclerosis have deceased serum BDNF level and increased VEGF level, suggesting that low level of BDNF and high level of VEGF may be involved in the occurrence and progression of carotid atherosclerosis.

HIV-1 infection changes miRNA expression profile in the whole blood / 病毒学报

To describe the unique miRNA profiles for HIV seropositive individuals and identify significantly differently expressed miRNAs, we determined the expression level of 754 miRNAs of 10 HIV seropositive individuals and 10 HIV seronegative individuals by using the Taqman low density microRNA array. BRB-Array Tool was used to conduct the significance analysis, and the DIANA online tool was used to perform the miRNA target prediction and pathway analysis. A total of 56 significantly differentially expressed miRNAs were identified by microarray between HIV seropositive and seronegative individuals. Among them, 49 miRNAs were down-regulated and 7 were up-regulated, partially overlapped with reported data. Predicted target genes were mainly involved in MAPK, TGF-beta and Wnt pathways. The results shows that miRNA profile changes in HIV-1 seropositive individuals, and the 56 differentially expressed miRNAs may play important role during HIV infection. Further studies on these miRNAs may be helpful for identify key molecules involved in HIV infection and potential diagnostic markers.

Immunogenicities and comparison of DNA vaccines encoding pol genes derived from B`/C and A/E recombinant HIV-1 strains / 中华预防医学杂志

<p><b>OBJECTIVE</b>To construct and compare the immunogenicities of DNA vaccines expressing pol genes derived from B`/C and A/E recombinant subtypes of HIV-1 in China.</p><p><b>METHODS</b>Two DNA vaccines were constructed by inserting the codon optimized pol genes derived from B'/C and A/E subtypes of HIV-1 into mammalian expression vector pSV1.0. In vitro expression efficiencies of the two DNA vaccines were determined by Western blotting and their immunogenicities were compared by i.m. immunizing female BALB/c mice. After immunization, mice splenocytes were isolated sterilely and IFN-γ based enzyme linked immunospot assay (ELISPOT) was employed to read out the specific T cell immunity.</p><p><b>RESULTS</b>The constructed DNA vaccines were validated by restriction enzyme digestion and DNA sequencing. Western blotting result showed both of the two DNA vaccines could be expressed at appreciable levels in vitro. Under the stimulation of Consensus B Pol peptide pools, specific T cell frequency elicited by pSVAE-Pol was (636±178) SFCs/10(6) splenocytes; specific T cell frequency elicited by pSVCN-Pol was (468±265)SFCs/10(6) splenocytes (P=0.412). Under the stimulation of HIV-1 AE2f Pol peptide pools, specific T cell frequency elicited by pSVAE-Pol was (1378±611) SFCs/10(6) splenocytes; specific T cell frequency elicited by pSVCN-Pol was (713±61) SFCs/10(6) splenocytes (P=0.134). Further analysis suggested pSVAE-Pol induced specific T cell responses mainly focused on Pol 1 peptide pool, while, in addition to induce Pol 1 specific T cell responses, pSVCN-Pol could also elicit T cell responses against consensus B Pol 2 peptide pool.</p><p><b>CONCLUSION</b>Although pSVAE-Pol was more immunogenic, pSVCN-Pol could induce T cell responses against broader epitope spectrum. Rational vaccine design may need combine them together.</p>

Altered regional homogeneity in spontaneous cluster headache attacks: a resting-state functional magnetic resonance imaging study / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Functional neuroimaging study has opened an avenue for exploring the pathophysiology of cluster headache (CH). The aim of our study was to assess the changes in brain activity in CH patients by the regional homogeneity method using resting-state functional magnetic resonance imaging technique.</p><p><b>METHODS</b>The functional magnetic resonance imaging scans were obtained for 12 male CH patients with spontaneous right-sided headache attacks during "in attack" and "out of attack" periods and 12 age- and sex-matched normal controls. The data were analyzed to detect the altered brain activity by the regional homogeneity method using statistical parametric mapping software.</p><p><b>RESULTS</b>Altered regional homogeneity was detected in the anterior cingulate cortex, the posterior cingulate cortex, the prefrontal cortex, insular cortex, and other brain regions involved in pain processing and modulation among different groups.</p><p><b>CONCLUSION</b>It is referred that these brain regions with altered regional homogeneity might be related to the pain processing and modulation of CH.</p>

The comparative analysis associated the postoperative ankle hole morphological changes of pilon fracture with the ankle function / 中华骨科杂志

ObjectiveTo investigate the relationship between the imaging manifestations and postoperative efficacy scores through measuring postoperative ankle mortise morphometry of Rüedi-Allg(o)wer Ⅲ pilon fracture.MethodsForty-seven cases of Rüedi-Allg(o)wer Ⅲ pilon fracture patients,including 20 males and 27 females with a mean of 43 years (range,21-65 years) were treated and followed up.Indexes including the width,height,depth,coronal angle and sagittal angle of ankle mortise were measured at the last follow-up.According to the Mazur score at the last follow-up,operated ankle functions were evaluated and divide into three groups (excellent group,good group and poor group.By comparing the five indexes of 47 cases and 3 groups between injury ankle and healthy ankle,we analyzed the correlation of the ankle mortise change and the ankle function.ResultsAll patients were followed up 18-24 months(mean,21 months).The results showed there were significant differences about all the measuring indexes except the height of ankle mortise between the injury ankle and healthy ankle of 47 follow-up cases.About the difference between both sides in 3 groups,the results showed there was a negative correlation between all indexes except the height of ankle mortise and the Mazur score.No correlation was found between the height of ankle mortise and the Mazur score.Recovering the ankle mortise's width,depth,coronal angle as well as sagittal anglein the surgery of severe pilon fractures has a significant effect on ankle function.ConclusionRecovery of the ankle mortise's width,depth,coronal angle as welld as sagittal angle during the surgery of severe pilon fractures has a significant effect on ankle function,so the anatomy of the ankle mortise should be recovered as much as possible.

Research progress of the anti-HIV activity of SAMHD1 / 病毒学报

The SAM and HD domain containing protein 1 (Sterile alpha motif domain and HD domain-containing protein 1, SAMHD1) is a putative negative regulator of the antiviral innate immune response. It can significantly increase the antiviral immune response, mediates the interferon-induced inflammatory response involved in the host foreign-virus defense system. The early studies have focused on its gene mutations associated with Aicardi-Goutières syndrome (AGS), the latest study found that SAMHD1 as a potent dGTP-stimulated triphosphohydrolase restricts HIV-1 replication by hydrolyzing the majority of cellular dNTPs, thus inhibiting reverse transcription and viral complementary DNA (cDNA) synthesis. Auxiliary gene of HIV-2 and simian immunodeficiency virus (SIVsm / mac) encoding the Vpx protein can eliminate HIV-1 restriction. In recent years, the research on SAMHD1, mores forward rapidly this paper overviews the recent research progression related to the above fields.

Cross-genome HIV-specific cytotoxic T-lymphocyte responses among HIV-1 infected individuals with varied infection time and routes / 中华检验医学杂志

Objective To investigate and compare the features of the HIV-1-specific CTL responses among three HIV-infected groups with varied infection history. Methods Three HIV-infeeted groups were enrolled in this study, including two groups infected by blood transmission (one group has been infected for more than 10 years and the other for 1-2 years) and one group of the man who have sex with man. The HIV-1-specific CTL responses were quantified by an IFN-γ based ELISPot assay with a peptide matrix system containing overlapping peptides spanning the entire HIV-1 Clade B genomic consensus sequences. Results The responding rate of CTL responses against all 17 peptide pools among the group that infected 1-2 years,the group infected more than 10 years and the group of MSM were 40% ,65% ,23%. One way ANOVO analysis showed that the responding rate of CTL responses against all 17 peptide pools were statistical significant among the three groups (F=19.96, P＜0.01);the magnitude of CTL responses of the three groups were 0-5 835 SFCs/106 PBMC, 0-7 225 SFCs/106PBMC, 0-9 740SFCs/106pBMC, Kruskal-Wallis test showed that the magnitude of CTL responses were statistical significant among the three groups( H = 101.90 , P ＜0.01);the breadth of CTL were 7 ( 2-11 ), 11(9-14) and 4 (2-6) respectively and Kruskal- Wallis test showed that the breadth of CTL had no statistical significant among the three groups( H = 34. 75 ,P ＜0. 01 ). The sequence of responding rate, magnitude and breadth of CTL from high to low was the group that had been infected for more than 10 years, the group infected 1-2 years and the sex transmission group. The common characteristics of the CTL response among the three groups were that the responding rate and the magnitude of the peptide Nef and Gag was higher than other peptide's. The magnitude of CTL responses among three different CD4count groups (CD4 ＜ 200/μl, CD4 200-500/μl, CD4 ≥500/μl,) was 0-18 475 SFCs/106pBMC, 350-34 095 SFCs/106pBMC, 490-21 550 SFCs/106 PBMC and had no statistic difference among the three different CD4 groups(H=2.93, P=0.23) while the breadth of CTL was 3(0-8), 10(2-17), 10 (1-17)respoctively and the breadth of CTL was lower in the group of CD4 count less than 200/μl than the other two groups( H = 14. 72, P ＜ 0. 01 ). The magnitude of CTL responses among three different viral load (VL)groups (VL＜ LDL, LDL ＜ VL ＜ 1 × 104 copys/ml, VL≥1 ×104 copys/ml) was 490-18 475 SFCs/106pBMC, 0-24 115 SFCs/106pBMC, 770-34 095 SFCs/106 pBMC and had no statistic difference among the three different viral load groups ( H = 0.79, P=0.67) and the breadth of the three different viral load groups CTL was 8( 1-17), 11 (0-17), 8 (1-16) and Kruskal-Wallis test showed that there was no statistic difference among the three different viral load groups (H =5.27, P =0. 07). Conclusions All groups predominantly develope T cell immune responses against Nef and Gag proteins. With the elapse of HIV infection, the CTL responses are increased in both magnitude and responding rate. This information is important for vaccine development.

Immunogenicity of DNA vaccines encoding regulatory/accessory proteins derived from three different prevalent strains in China / 中华微生物学和免疫学杂志

0bjective To determine the immunogenicities of DNA vaccines expressing tat-rev-integrase(c-half)-vif-neffusion genes(TRIVN) derived from prevalent B', B'/C and AE recombinant subtypes of HIV-1 in China. Methods Two DNA vaccines were constructed by inserting the codon optimized tat-revintegrase(c-half)-vif-nef fusion genes derived from B' and B'/C subtype of HIV-1 into mammalian expression vector pSVI. 0. DNA vaccine containing tat-rev-integrase (c-half)-vif-nef fusion gene derived from HIV-1AE2f has been constructed previously. In vitro expression efficiencies of three DNA vaccines were determined by Western blot and their immunogenicities were compared by immunizing female BALB/c mice. IFN-γ ELISPOT assay was used to read out the specific T cell immunity. Results The constructed DNA vaccines were validated by restriction enzyme digestion and DNA sequencing. Western blot assay showed three constructed DNA vaccines could be expressed at a comparable level in vitro. After vaccination, AE-TRIVN mounted T cell immune responses at (948.0 ± 330.0) SFCs/106 splenocytes, followed by the mixed DNA vaccine[ (500.0 ± 155.0) SFCs/106 splenocytes ], RL-TRIVN r[ ( 195. 1 ± 44.0) SFCs/106 splenocytes ]and CN-TRIVN [ (89.5 ± 17.0) SFCs/106 splenocytes]. Interestingly, we observed that single DNA vaccination induced specific T cell responses predominantly targeting Integrase (C-half) and Vif, whereas the mixed DNA could significantly improve T cell responses against Nef. Conclusion AE-TRIVN was the most immunogenic among the three DNA vaccines and the mixed DNA vaccination could change the immunogenic hierarchy of T cell epitopes across the fusion genes vaccine.

HIV-specific IL-2(+) and/or IFN-γ(+) CD8(+) T cell responses during chronic HIV-1 infection in former blood donors / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Conflicting data have been generated from previous studies to determine which kind of relationship exists between HIV-1 specific CD8 Tcell responses and HIV-1 viral load or CD4 count over the course of infection. In this study, 153 HIV-1 infected LTNPs were enrolled to investigate the role of HIV-1 specific CD8 T-cell responses in chronic HIV-1 infection among HIV-1 infected former blood donors.</p><p><b>METHODS</b>The patients were stratified into three groups according to CD4 count: CD4≥500 cells/μL; 350 cells/μL≤CD4<500 cells/μL; CD4<350 cells/μL. PBMCs were isolated from the patients' anticoagulated blood samples. IL-2 and IFN-γ secretions of CD 8 T cells against 17 HIV-1 consensus B full peptide pools were analyzed by using ICS assay.</p><p><b>RESULTS</b>An overall inverse correlation were observed between CD4 count and plasma viral load. Although no significant difference was observed during the comparisons of frequency/breadth of HIV-1 specific CD8 T cell responses, CD4 count stratification analysis showed that different correlation pattern existed in three strata: as for patients whose CD4 counts were less than 350 cells/μL, no significant correlations were identified between frequency/breadth of HIV-1 specific CD8 T cell responses and CD4 count/viral load; as for patients whose CD4 counts ranged from 350 cells/μL to 500 cells/μL, significant correlation was only observed between the response breadth of IL-2+IFN-γ+ CD8 T cells and CD4 count; however, as for patients whose CD4 counts were more than 500 cells/μL, direct correlations were identified between IL-2+IFN-γ+/IL-2+/IFN-γ+ CD8 T cells and viral load or CD4 count.</p><p><b>CONCLUSIONS</b>Universal consistent inverse correlation was only indentified between CD4 count and viral load. The relationship between HIV-1 specific CD8 T cell responses and CD4 count/viral load varied in different CD4 strata, which showed that better preserved CD4 T cells were correlated with better CD8 T cell functions.</p>

Establishing a Th17 based mouse model for preclinical assessment of the toxicity of candidate microbicides / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>To effectively block the invasion of human immunodeficiency virus (HIV)-1 on mucosal surface, vaginal anti-HIV-1 microbicides should avoid inflammatory responses and disruption of mucosa integrity because these will facilitate transepithelial viral penetration and replication. However, existing models fail to predict and evaluate vaginal mucosal toxicity induced by microbicides, and most importantly, they are unable to identify subtle or subclinical inflammatory reactions. This study was designed to develop a cost-effective in vivo model to evaluate microbicide safety in a preclinical study which can recapitulate the mucosal topical reaction.</p><p><b>METHODS</b>A murine model was employed with nonoxynol-9 (N-9) as the topical stimulant within the vagina. Different concentrations of N-9 (1%, 3%, and 4%) were topically applied to the vagina for five consecutive days. A panel of inflammatory cytokines including interleukine-2 (IL-2), IL-4, IL-6, IL-17A, interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and immuno-regulatory IL-10 were assayed in vaginal lavage. Cytokines were quantified by using cytometric bead array (CBA) and reverse transcript (RT) real-time PCR. Histopathological evaluation of vaginal tissues was conducted on hematoxylin-eosin stained slides and scored with a semi-quantitative system according to the severity of epithelial disruption, leucocyte infiltration, edema, and vascular injection. The association between the cytokines and histopathological scores was assessed by linear regression analysis.</p><p><b>RESULTS</b>All three concentrations of N-9 induced inflammatory cytokine production. The 4% N-9 application resulted in a consistent production of cytokines in a time-dependent manner. The cytokines reached peak expression on day three with the exception of IL-4 which reached its peak on day one. Histopathological examination of 4% N-9 treated cervicovaginal tissues on day three showed intensive damage in four mice (sores: 10 - 13) and moderate damage in one mouse (score: 8), which were significantly associated with both inflammatory cytokines IL-17A and IL-6 and anti-inflammatory cytokines IL-4 and IL-10. Interestingly, IL-17A showed significant positive association with inflammatory cytokine TNF-α (r = 0.739; P < 0.05), anti-inflammatory cytokines IL-10 (r = 0.804; P < 0.01) and IL-4 (r = 0.668; P < 0.05).</p><p><b>CONCLUSIONS</b>Our data demonstrate that a panel of cytokines (IL-17A, IL-6, IL-4 and IL-10) could be used as surrogate biomarkers to predict the histopathological damage. Th17 may play a central role in orchestrating inflammatory cytokine responses. This Th17 based mouse model is cost-effective and suitable to assess the toxicity of candidate microbicides in preclinical studies.</p>

Immunogenicity of DNA vaccines encoding structural proteins and regulatory/accessory proteins derived from an HIV-1 CRF01_AE isolate circulating in China / 中华微生物学和免疫学杂志

Objective To construct two DNA vaccines encoding Gag-Env fusion protein and Tat-Rev-Integrase(C-half)-Vif-Nef fusion protein derived from the first HIV-1 CRF01_AE isolate(AE2f) in Chi-na and to evaluate the immunogenicity in mice. Methods Two DNA vaccines were constructed by inserting the codon optimized and synthesized gag-env fusion gene and tat-rev-integrase(c-half)-vif-nef fusion gene de-rived from AE2f into mammalian expression vector pDRVISV1. 0, the generated DNA vaccines were desig-nated as pSVAE/GE and pSVAE/TRIVN, respectively, and their in vitro expression were determined by Western blot with transfected 293T cells. Mice were i. m. immunized with either pDRVI1.0 as mock control, pSVAE/GE or pSYAE/TRIVN for 4 times at two-week interval. Two weeks following the final im-munization, cellular responses to pool of HIV-1 Env, Gag, Tat, Rev, Intergrase, Vif and Nef peptides were evaluated by ELISPOT assay. Results The construction of DNA vaccine pSVAE/GE and pSVAE/TRIVN was validated by restriction enzyme digestion and bidirectional sequencing. Western blot showed a specific band at molecular mass 220×10~3 in lane of pSVAE/GE transfeeted 293T cell and a specific band at 95×10~3 in the lane of pSVAE/TRIVN. Both DNA vaccines mounted significant specific T cell responses with (3010 ± 566) SFC/10~6 splenocytes for DNA vaccine pSV AE/GE and (948 ± 737) SFC/10~6 spleno-cytes for DNA vaccine pSVAE/TRIVN, whereas the mock control of pDRVISV1.0 only raised marginal T cell responses. Conclusion Both pSVAE/GE and pSVAE/TRIVN were capable of expressing the inserted fusion immunogen genes and able to elicit vigorous cellular immune responses, therefore, these DNA vac-cines are highly immunogenic.

A mouse model based on replication-competent Tiantan vaccinia expressing luciferase/HIV-1 Gag fusion protein for the evaluation of protective efficacy of HIV vaccine / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Developing an effective vaccine against human immunodeficiency virus type 1 (HIV-1) remains a grand challenge after more than two decades of intensive effort. It is partially due to the lack of suitable animal models for screening and prioritizing vaccine candidates. In this study, we aim to develop a mice model to test HIV-1 vaccine efficacy.</p><p><b>METHODS</b>We constructed a recombinant vaccinia expressing firefly luciferase and HIV-1 Gag fusion protein based on Tiantan strain, an attenuated but replication-competent poxvirus (rTTV-lucgag). By quantifying the luciferase activity as its read out, we defined the biodistribution of Tiantan strain poxvirus in mice inoculated intraperitoneally and attempted to apply this model to evaluate the HIV-1 vaccine efficacy.</p><p><b>RESULTS</b>Our data demonstrated that the rTTV-lucgag was able to express high level of luciferase (< or = 10(6) relative luciferase units (RLU)/mg protein) and HIV-1 Gag (> 3 folds increase comparing to the control). After intraperitoneal inoculation, this virus had dominant replication in the ovary, uterus, and cervix of mice and the luciferase activities in those organs are significantly correlated with viral titers (r(2) = 0.71, P < 0.01). Pre-immunization with an HIV gag DNA vaccine reduced the luciferase activity in ovary from (6006 +/- 3141) RLU/mg protein in control group to (1538 +/- 463) RLU/mg protein in vaccine group (P = 0.1969).</p><p><b>CONCLUSIONS</b>The luciferase activity in ovary could represent viral replication in vivo; this rTTV-lucgag/mice model may be suitable to assess the protective efficacy of cytotoxic T-cell responses to HIV Gag with less tedious work and high through-put.</p>