[Analysis on the relationship of molecular transmission between HIV infected men who have sex with men and their sexual partners].

Objective: To investigate the relationship between HIV-infected men who have sex with men and their sexual partners in Zhejiang province. Methods: A cross-sectional survey was conducted to recruit newly confirmed HIV/AIDS among MSM from 2015 to 2017, including sexual partner identification and molecular epidemiological study. Plasma was collected to extract RNA, and the pol gene of HIV-1 was amplified by RT-PCR/nested PCR. Phylogenetic tree and molecular transmission cluster were analyzed to identify the transmission relationship between sexual partners. Results: A total of 937 HIV/AIDS among MSM were recruited to promote HIV testing for their sexual partners, and 173 positive sexual partners were identified. 50.8% (61/120) of the gene sequences were clustered among the positive sex partners. Seven pairs of clustered sex partners combined with the results of recent infection preliminarily determined the transmission direction. In the clusters, there were statistical differences between the partners who were diagnosed in the same year (OR=12.190, 95%CI: 1.563-95.054) or with current residence in the different districts (OR=17.054, 95%CI: 1.742-166.982). Conclusions: Combined with a molecular transmission network, HIV test for the sexual partners of HIV/AIDS among MSM can improve the accurate tracking of cases and preliminarily determine the direction of transmission, according to the results of recent infection. It is suggested that after HIV is confirmed for HIV/AIDS among MSM, HIV tests should be carried out as soon as possible for their sexual partners, including a cross-regional sexual partner tracking test, which is helpful to improve the tracing procedure.

[Characteristics of subtypes and transmission of HIV-1 infected persons among student MSM in Zhejiang province, 2013-2015].

Objective: To investigate the characteristics of subtype diversity and transmission on HIV-1 among 12 to 30 years old student MSM in Zhejiang province. Methods: A total of 290 newly diagnosed HIV infected student MSM were selected as the research objects for molecular studies on HIV, in Zhejiang province during 2013 to 2015. Data on epidemiology and plasma samples of these people were collected. HIV-1 nucleotide sequences of pol gene regions were amplified using the RT-PCR/nested PCR method and sequenced. Phylogenetic analysis was performed to determine the HIV-1 genotypes. Characteristics of transmission mode among these cases were also analyzed. Results: A total of 290 cases, 50.3% were diagnosed in Hangzhou and 81.0% had college or above degrees. 178 sequences including 10 subtypes, were obtained, with the main subtypes as CRF01_AE (49.4%, 88/178) and CRF07_BC (39.3%, 70/178). A total of 18 molecular transmission clusters were formed (42 cases, cluster size from 2 to 4), with the proportions of clusters as 23.6% (42/178). 61.9% (26/42) of student MSM with their schools located in the same district within the transmission clusters. Their sexual partners would include both student MSM and non-student MSM. The proportion of clusters among middle school students was 38.2% (13/34), higher than that of college students (20.1%, 29/144) ( χ(2)=4.996, P<0.05). Conclusions: The HIV-1 subtypes of student MSM in Zhejiang province appeared diversity, which indicated with the diversity of sources of infection. The geographical distribution of cluster cases is relatively centralized. In order to effectively control the spread of AIDS, more attention should be paid to the sexual partners involved and to specific programs on intervention.

[Retrospective cohort study on the correlation between high value of lactic acid and risk of death in 127 patients with extensive burn during shock stage].

Objective: To analyze the relationship between serum lactic acid value and risk of death in patients with extensive burn during shock stage and the related influencing factors. Methods: Clinical data of 127 patients (111 males and 16 females) with extensive burn admitted to Institute of Burn Research of the First Affiliated Hospital of Army Medical University from January 2009 to December 2013 and Department of Plastic Surgery and Burns of the Affiliated Hospital of Southwest Medical University from January 2016 to December 2018, who met the admission criteria, were retrospectively analyzed. The patients aged 21 to 62 years, with total burn area more than 50% total body surface area. All patients were treated with antishock therapy after admission. (1) According to the treatment outcome, the patients were divided into survival group (n=98) and death group (n=29). The gender, age, total burn area, partial-thickness burn area, full-thickness burn area, abbreviated burn severity index (ABSI), admission time after injury, number of patients with inhalation injury, number of patients with acute renal failure, and serum lactic acid values on admission and at post admission hour (PAH) 12, 24, 36, and 48 were recorded. (2) According to the optimal positive cut-off value of serum lactic acid 48 hours after admission, the patients were divided into high lactic acid group and normal lactic acid group. Age, gender, total burn area, indexes at PAH 48 including urea nitrogen, creatinine, alanine aminotransferase (ALT), aspartate aminotransferase (AST), total serum bilirubin, alkaline phosphatase (ALP), albumin, white blood cell count, platelet count, lymphocyte count, prothrombin time (PT), hematocrit value, oxygenation index, respiratory index (RI), the alveolar-arterial oxygen partial pressure difference, mean arterial pressure (MAP) at PAH 48, the average urine volume within 48 hours after admission, the total volume of intravenous fluid infusion within 48 hours after admission, the volume of fluid infusion per kilogram of body mass within the first 24 hours after admission, the volume of fluid infusion per one percent of body surface area per kilogram of body mass within the first 24 hours after admission, the volume of urine per kilogram of body mass per hour within the first 24 hours after admission, and the percentage of hospital death were recorded. Data were processed with t test, chi-square test, and Fisher's exact probability test. Cox regression analysis was used to screen independent risk factors affecting the prognosis of patients. Receiver operating characteristic curve (ROC) of serum lactic acid value at PAH 48 of 127 patients was drawn to predict patients' death and determine the optimal positive cut-off value. Multivariate logistic regression analysis was used to screen independent risk factors causing increase of serum lactic acid. Results: (1) There were significantly statistical differences in total burn area, full-thickness burn area, and ABSI of patients between survival group and death group (t=6.257, 4.476, 5.727, P<0.01), while other indexes between the two groups were close. (2) The serum values of lactic acid of patients in death group on admission and at PAH 12, 24, 36, and 48 were (4.00±0.28), (4.50±0.26), (4.02±0.31), (3.48±0.22), (3.40±0.19) mmol/L, respectively, which were significantly higher than those in survival group [(3.30±0.21), (3.20±0.19), (2.33±0.17), (1.85±0.18), (1.50±0.09) mmol/L, t=14.552, 29.603, 38.133, 40.648, 74.973, P<0.05 or P<0.01]. (3) Cox regression analysis showed that the serum value of lactic acid at PAH 48 was the independent risk factor affecting the prognosis of patients, with risk ratio of 1.853 and 95% confidence interval of 1.342-2.559, P<0.01. (4) The total area under ROC of serum value of lactic acid at PAH 48 to predict death of 127 patients was 0.811, with 95% confidence interval of 0.699-0.924, P<0.01. The optimal positive cut-off value of serum value of lactic acid was 1.75 mmol/L, with sensitivity of 75.0% and specificity of 79.5% for predicting death. (5) There were significantly statistical differences in total burn area, ALT, AST, ALP, PT, total serum bilirubin, total volume of intravenous fluid infusion within 48 hours after admission, volume of fluid infusion per kilogram of body mass within the first 24 hours after admission, and percentage of hospital deaths of patients between high lactic acid group (n=34) and normal lactic acid group (n=93), t=3.592, 6.797, 10.367, 2.089, 2.880, 4.517, 2.984, 4.044, χ(2)=58.498, P<0.05 or P<0.01, while other indexes were close between the two groups. (6) Multivariate logistic regression analysis showed that AST and total serum bilirubin were independent risk factors for increase of serum lactic acid, with odds ratios of 1.021 and 1.064 and 95% confidence intervals of 1.001-1.040 and 1.001-1.132, P<0.05. Conclusions: Serum value of lactic acid at PAH 48 can independently predict the death of patients with extensive burns. Liver injury is an important risk factor causing hyperlacticemia during burn shock stage. Widespread increase of vascular permeability and large amount of fluid resuscitation are the core factors leading to aggravation of abdominal organ injury.

[Characteristic analysis of molecular subtypes and recombinant structure of HIV-1 infection in Zhejiang Province, 2015].

Objective: To understand the molecular subtypes and recombinant structure of HIV-1 infection in Zhejiang Province. Methods: According to the method of stratified random sampling method, a total of 302 newly HIV infected people (excluding AIDS patients) were selected as the subjects of the investigation from April to June, 2015, who were all newly HIV positive patients reported in Zhejiang Province and were diagnosed as HIV positive and had not received antiretroviral treatment. Questionnaires were used to collect information such as gender, residency, marital status, route of transmission and case reporting area. At least 8 ml of venous blood was collected from HIV infected people, reverse transcription-polymerase chain reaction (RT-PCR) and nested PCR were used to amplify the pol gene sequence of HIV-1 strains and circulating recombination forms (CRF) and unique recombinant form (URF) strains were screened using phylogenetic tree, and molecular subtype analysis and recombinant analysis were performed. Results: Of the 302 HIV infected people, 18 cases were infected with new CRFs or URFs, among which 15 were male (83.3%), 3 were heterosexual transmission (17.7%); 8 cases (44.4%) were infected with new CRFs, which included CRF55_01B 5(27.8%), CRF67_01B 2 (11.1%), CRF69_01B 1 (5.6%); 10 cases (55.6%) were infected with URFs, in which 6 cases were infected with two groups of recombinant strains composed of CRF01_AE subtype and CRF07_BC subtypes in pol region, 3 cases were infected with three groups of recombinant strains of the two subtypes in pol region, and 1 case could not be accurately determined of the recombinant structure. The results of recombinant source analysis showed that the newly recombinant strains found from Zhejiang province were highly homologous to the strains from Guangdong, Anhui, Hebei, Yunnan, Liaoning province, and the sequence similarity was 94.4%-98.0%. Conclusion: The new recombinant strains of Zhejiang province included CRFs strains and URFs strains with CRF01_AE and CRF07_BC as the main recombinant structure mode, and all of them were introduced from other places and formed a trans-regional chain of HIV transmission.

[Analysis on probability of premature death and cause eliminated life expectancy of major non-communicable diseases in Chongqing Municipality, 2016].

Objective: To analyze the premature death probability and cause-eliminated life expectancy of cardiovascular disease, cancer, chronic respiratory disease and diabetes in Chongqing residents in 2016 so as to provide recommendation for non-communicable diseases (NCDs) prevention and control in Chongqing. Methods: Death cases of Chongqing Municipality between January 1(st) and December 31(st), 2016 were reported through death case registry system of national center for disease prevention and control. Death cases were sorted by international classification of disease (ICD-10). Mortality rate, standardized mortality rate, constituent ratio, premature death probability, life expectancy, and cause-eliminated life expectancy of four major NCDs were analyzed. Results: A total of 218 004 death cases were reported in Chongqing, 2016, and the mortality rate was 731.73/100 000. Of them, a total of 179 637 death cases of the four major NCDs including cardiovascular disease, cancer, chronic respiratory disease and diabetes were reported, accounting for 82.40% of all death cases. The mortality rate and standardized mortality rate of four major NCDs was 602.95/100 000 and 455.82/100 000, respectively. The premature death probability of four major NCDs was 15.96%, and males (25.39%) had a higher premature death probability than females (10.78%). The premature death probability of cardiovascular disease, cancer, chronic respiratory disease, and diabetes were 6.01%, 8.32%, 2.05%, and 0.43%, respectively. Life expectancy would increase by 6.02, 3.19, 1.89, and 0.19 years, after eliminating cardiovascular disease, cancer, chronic respiratory disease and diabetes respectively. Conclusion: The premature death probability of major NCDs was high in Chongqing, and males had a higher premature death probability than females did. Intervention and health management of the population should be conducted according to different gender-based risk factors to reduce the premature death probability.

An atlas and analysis of bovine skeletal muscle long noncoding RNAs.

Long noncoding RNAs (lncRNAs) have various biological functions and have been extensively studied in recent years. However, the identification and characterization of bovine lncRNAs in skeletal muscle has been very limited compared with that of lncRNAs in other model organisms. In this study, 7188 bovine skeletal muscle lncRNAs were identified by RNA-Seq and a stringent screening procedure in four different muscle tissues. These lncRNAs shared many characteristics with other mammalian lncRNAs, such as a shorter open reading frame and lower expression level than for mRNAs. Furthermore, the chromosomal locations and global expression patterns for these lncRNAs are also described in detail. More importantly, we detected the important interaction relationships of lncRNAs-miRNAs-mRNAs related to muscle development among 36 lncRNAs, 62 miRNAs and 12 mRNAs. Our results provide a global expression pattern of lncRNAs specific to bovine skeletal muscle and provide important targets for revealing the function of bovine muscle development by thoroughly studying the interaction relationships of lncRNAs-miRNAs-mRNAs.

[Analysis on HIV-1 subtypes and transmission clusters in newly reported HIV/AIDS cases in Yiwu, Zhejiang Province, 2016].

Objective: To understand the characteristics of distribution on HIV-1 subtypes and the transmission clusters in Yiwu in Zhejiang province. Methods: A cross-sectional study of molecular epidemiology was carried out on newly reported HIV/AIDS cases in Yiwu. RNA was extracted from 168 plasma samples, followed by RT-PCR and nest-PCR for pol gene amplification, sequencing, phylogenetic tree construction used for analyzing the subtypes and transmission clusters. Mutations on drug resistance was analyzed by CPR 6.0 online tool. Results: Subjects were mainly males (86.3%, 145/168), with average age as (39.1±13.4) years old and most of them were migrants (66.7%, 112/168). The major routes of transmission included homosexual (51.2%, 86/168) and heterosexual (48.8%, 82/168) contacts. The rate of success for sequence acquisition was 89.9% (151/168). The dominant subtypes showed as CRF01_AE (74, 49.0%) and CRF07_BC (64, 42.4%), followed by CRF08_BC (5, 3.3%), CRF55_01B (3, 2.0%), each case of subtype B, CRF45_cpx, CRF59_01B, CRF85_BC and URF (B/C). CRF45_cpx and CRF85_BC were discovered the first time in Zhejiang province. Twenty-six transmission clusters involving 65 cases were found, with the total clustered rate as 43.0% (65/151), in which the CRF01_AE clustered rate appeared as 54.1% (40/74), higher than that of CRF07_BC (21/64, 32.8%). The average size of cluster was 2.5 cases/cluster, with average size of cluster in CRF01_AE patients infected through heterosexual transmission as the largest (3.5 cases/cluster). The prevalence of transmitted drug resistance was 4.6% (7/151). Seven cases with surveillance drug resistant mutations (SDRM) were found, including 5 cases of M46L (3.3%), and one case of F77L or Y181C. Conclusion: HIV genetic diversity and a variety of transmission clusters had been noticed in this study area (Yiwu). Programs on monitoring the subtypes and transmission clusters should be continued and strengthened.

Promoter analysis of bovine cardiomyopathy-associated protein 1 gene.

The CMYA1 (cardiomyopathy-associated protein 1) is an actin-binding protein that plays a vital role in cardiac morphogenesis. CMYA1 is expressed specifically in the myocardial and skeletal muscle and is up-regulated in injured muscle. We therefore speculated that the bovine CMYA1 promoter might be muscle-specific. In this study, the promoter (+20/-1135) region of the bovine CMYA1 gene was cloned into a pEGFP-1 vector, and we found that the EGFP was observed only in C2C12 and myoblast cells. Thus, the CMYA1 promoter is muscle-specific. Thereafter, eight pGL3-basic vectors with various truncated CMYA1 promoter fragments were transfected into C2C12 cells, to identify the core promoter region using a Dual-Luciferase Reporter Assay System. The results showed that the promoter region from -457 to +20 bp was essential for CMYA1 to maintain the promoter activity, implying that this region may be the CMYA1 core promoter. We thus illustrate that the core promoter is muscle-specific. To evaluate the activity of the CMYA1 core promoter, the CMYA1 core and muscle creatine kinase (MCK) promoters were cloned into a pcDNA3.1 vector. The expression levels of their target genes were measured in C2C12 cells using real-time polymerase chain reaction. The CMYA1 promoter drove the expression of the target gene six times higher than did the MCK promoter. The results thus suggest that the CMYA1 promoter could be an effective muscle-specific promoter, which may be useful in further studies of cardiomyopathy treatment and transgenic animal research.

Selection of reference genes in canine uterine tissues.

Real-time quantitative polymerase chain reaction (RT-qPCR) is usually employed in gene expression studies in veterinary research, including in studies on canine pyometra. Canine pyometra is a common clinical disease in bitches. When using RT-qPCR, internal standards, such as reference genes, are necessary to investigate relative gene expression by quantitative measurements of mRNA levels. The aim of this study was to evaluate the stability of reference genes and select reference genes suitable for canine pyometra studies. We collected 24 bitch uterine tissue samples, including five healthy and 19 pyometra infected samples. These were used to screen the best reference genes of seven candidate genes (18SrRNA, ACTB, B2M, GAPDH, HPRT, RPL13A, and YWHAZ). The method of KH Sadek and the GeNorm, Normfinder, BestKeeper, and RefFinder software were used to evaluate the stability of gene expression in both pyometra and healthy uterine samples. The results showed that the expression stability of the candidate gene in pyometra and healthy tissues differed. We showed that YWHAZ was the best reference gene, which could be used as an accurate internal control gene in canine pyometra studies. To further validate this recommendation, the expression profile of a target gene insulin-like growth factor 1 receptor gene (IGF1R) was investigated. We found that the expression of IGF1R was significantly altered when different reference genes were used. All reference genes identified in the present study will enable more accurate normalization of gene expression data in both pyometra infected and healthy uterine tissues.

[Analysis on antiretroviral therapy efficacy for HIV/AIDS in Zhejiang province, 2009-2014].

OBJECTIVE: To analyze the efficacy of highly active antiretroviral therapy(HAART)for HIV/AIDS in Zhejiang province from 2009 to 2014. METHODS: A retrospective cohort study was conducted among patients with access to HAART aged <15 years from 2009 to 2014 through descriptive epidemiological analysis on the patients' demographic characteristics, HAART efficacy. The immune recovery Kaplan-Meier curves were drawn by using baseline CD(4) T lymphocytes cell(CD(4))data. The influencing factors for virus suppression were analyzed by using logistic regression model. RESULTS: There were 9 594 patients with access to HAART in Zhejiang from 2009 to 2014. The' average age of the patients was 38.7 years old, the male to female ratio was 4.11∶1. Heterosexual(55.9%)and homosexual(40.0%)contacts were the main transmission routes. Baseline CD(4) count 200-349 cells/µl was found in 45.4% of the patients, the median of the CD(4) count was 221(IQR: 123-298). The median of time for CD(4) recovery in patients with baseline CD(4) count <200 cells/µl was 3.48 and 11.58 times higher than the patients with baseline CD(4) count 200-349 cells/µl and baseline CD(4) count ≥350 cells/µl. Univariate and multivariate logistic regression analyses showed that marital status, HAART regimen, treatment miss in last week and CD(4) recovery were major influencing factors for HAART efficacy. CONCLUSIONS: HAART had good treatment effecacy in HIV/AIDS patients in Zhejiang. The baseline CD(4) count and adherence to HAART were the most important influencing factors. It is recommended to take measures to expand HIV screening and HAART coverage, conduct HAART adherence education, and strengthen the special care for high-risk groups to improve the HAART efficacy in Zhejiang province.

Overexpression of the A-FABP gene facilitates intermuscular fat deposition in transgenic mice.

Adipocyte fatty acid-binding protein (A-FABP), the most abundant FABP in adipocytes, controls fatty acid uptake, transport, and metabolism in fat cells. We constructed a transgenic mice model that overexpressed the cattle A-FABP gene to investigate the relationship between A-FABP expression and intermuscular fat deposition. There was no significant difference in body weight and serum biochemical indexes between transgenic and wild-type mice. Further, there were no significant differences in intermuscular triglyceride content and A-FABP expression levels over three generations of transgenic mice. However, abdominal adipose rate, A-FABP protein content, and intermuscular triglyceride levels of transgenic mice were significantly higher than those of wild-type mice. In addition, triglycerides were remarkably higher in the skeletal muscle but lower in the myocardium of transgenic mice. Thus, overexpression of cattle A-FABP gene promoted fat deposition in the skeletal muscle of transgenic mice.

Chestnutamide: a novel alkaloid from flowers of Castanea mollissima Blume.

A novel alkaloid, chestnutamide, was isolated from the flowers of Castanea mollissima Blume. The structure was determined on the basis of spectral analysis.