Objective To analyze the changes of liver and kidney function, blood glucose and lipid metabolism at different follow-up time points of different treatment regimens, and to provide reference for clinical optimization and adjustment of medication in HIV/AIDS patients. Methods The changes of liver and kidney function, blood glucose and lipid metabolism at seven follow-up time points were analyzed retrospectively. The baseline blood collection time of HIV /AIDS patients was set as the starting point, and the final follow-up time was set as the end point. The seven follow-up points were 0, 3, 6, 9, 12, 18 and 24 months respectively. Results There were statistically significant differences in the distribution of sex, age, education, marital status, WHO staging, infection route, and baseline CD4+T lymphocyte count among 605 enrolled patients based on different treatment regimens. Liver function: The level of T-Bil in group E was higher than that of baseline at 9M, 12M, 18M and 24M after treatment (P<0.01); In group F, the level of T-Bil was higher than that of baseline at 9M after treatment (P=0.001); The levels of ALT in group C at the six follow-up points after treatment were higher than the baseline (P<0.001); The level of AST in group C was higher than that of baseline after 3M and 6M treatment (P<0.05). Renal function: The level of UREA in group C was higher than that in baseline after 6M treatment (P=0.007); The level of UREA in group F was higher than that in the baseline after 12M treatment (P<0.001); The level of UA in group F was higher than that of baseline after 3M, 6M and 12M treatment (P<0.05). Blood lipid and blood glucose: The levels of Glu at some follow-up points after ART treatment in group A and group C were higher than that at baseline (P<0.05); The levels of TG at some follow-up points in group A, group E and group F after ART treatment were higher than those at baseline (P<0.05); The levels of TC at some follow-up points in group A, group B, group C, group E and group F after ART treatment were all higher than the baseline (P<0.05). Conclusion Regular monitoring of changes in laboratory indicators of different treatment regimens during ART is of great importance to the prognosis of patients. Different laboratory indicators should be monitored according to different treatment regimens to effectively prevent adverse reactions caused by different treatment regimens.

Objective:To investigate the molecular network of HIV-1 CRF01_AE in Yunnan Province and the factors influencing it.Methods:Demographic data and plasma samples of HIV/AIDS patients in Yunnan drug resistance monitoring database from 2018 to 2021 were collected. HIV-1 pol gene fragments (protease and reverse transcriptase region) were amplified using RT-PCR and then sequenced. The optimal gene distance was selected and a molecular network was constructed based on the sequences of CRF01_AE genotype. Results:In this study, a total of 967 sequences of CRF01_AE genotype were obtained by sequencing. At the optimal gene distance threshold of 1.75%, a total of 320 sequences were involved in the network with a rate of 33.1%, and 84 clusters were identified. In the regional distribution, one cluster dominated by multiple regions, one cluster dominated by Zhaotong, one cluster dominated by Honghe and five clusters dominated by Wenshan were formed in the network. In the network, 75.8% of heterosexual men were connected with other heterosexual men and 54.1% were connected with heterosexual women. There was potential transmission among 66.7% of men who have sex with men (MSM). HIV/AIDS patients in Chuxiong, Dali, Dehong, Honghe, Lincang, Pu′er, Wenshan, Yuxi and Zhaotong were more likely to be involved in the network that those in Kunming. People who were 50 years old and above were more likely to be involved in the network than those less than 25 years old. Factors influencing HIV/AIDS patients with HIV-1 CRF01_AE infection to become high-risk transmitters in Yunnan were not found and further study on this subject was needed.Conclusions:HIV-1 CRF01_AE strains had spread actively in different regions of Yunnan Province and the transmission network was complex. Dynamic monitoring of CRF01_AE strains should be strengthened and a precise intervention for high-risk transmitters should be performed to reduce new infections.

Objective:To explore the relationship between drug resistance occurrence and the distribution pattern of polymorphic loci in individuals with human immunodeficiency virus infection/acquired immunodeficiency syndrome (HIV/AIDS) treated with highly active anti-retroviral therapy (HAART).Methods:HAART-failed HIV/AIDS patients who successfully amplified the gene sequences of the pol region between June 2015 and December 2021 from 16 prefecture-level administrative regions in Yunnan Province were included.The resistant sequences were classified using the human immunodeficiency virus (HIV) basic local alignment search tool (BLAST) and validated through MEGA 6.0, and the obtained sequences were submitted to the Stanford University HIV Drug Resistance Database to identify drug resistance loci. The distribution of polymorphic loci was analyzed across patients exhibiting varying degrees of drug resistance, different treatment regimens and distinct HIV-1 subtypes.Changes of the frequencies of polymorphic loci in patients with different degrees of drug resistance were analyzed using trend chi-square test. Statistical comparisons and further paired comparisons were performed using chi-square test.Results:Gene sequences were amplified from 1 453 patients, and the resistance testing results showed 954 sensitive, 224 potentially or low resistant, 189 moderately resistant, and 86 highly resistant patients. The frequencies of mutations I15V, L19I, D60E in the HIV-1 protease region (PR region) and E36A, T39D, S48T mutations in the HIV-1 reverse transcriptase region (RT region) showed a decreasing trend as the degree of HIV-1 resistance escalated ( χ2trend=19.86, 9.16, 13.66, 37.64, 18.44 and 40.86, respectively, all P<0.01). Conversely, the mutations V77I in the PR region and K122E in the RT region showed an ascending trend ( χ2trend=12.19 and 10.03, respectively, both P<0.01). Distinct treatment groups, namely zidovudine (AZT)+ lamivudine (3TC)+ lopinavir/ritonavir (LPV/r), AZT+ 3TC+ efavirenz (EFV), AZT+ 3TC+ nevirapine (NVP), and tenofovir (TDF)+ 3TC+ EFV, were examined. Statistically significant differences in the frequencies of mutations E35D, M36I, and D60E in the PR region, as well as S48T, K122E, and R211K in the RT region, were observed among these treatment groups ( χ2=22.46, 9.32, 14.46, 26.85, 18.92 and 24.26, respectively, all P<0.05). In paired comparisons, AZT+ 3TC+ LPV/r group displayed higher frequencies of E35D, M36I, and D60E mutations, the AZT+ 3TC+ EFV group showed a higher frequency of S48T mutation, the AZT+ 3TC+ NVP group showed a higher frequency of K122E mutation, and the TDF+ 3TC+ EFV group exhibited a higher frequency of R211K mutation, all with statistically significant differences (all P<0.008). The differences in the frequencies of T12S, I15V, L19I, M36I, V77I, L89M in the PR region and E53D, I135V, S162C, R211K, K277R in the RT region among circulating recombinant form (CRF)08_BC, CRF07_BC and CRF01_AE subtype group were statistically significant ( χ2=693.60, 712.51, 798.11, 434.85, 386.91, 657.78, 932.58, 409.21, 344.39, 469.44 and 260.48, respectively, all P<0.001). In paired comparisons, the frequencies of T12S, I15V, L19I, E53D, I135V, S162C and R211K in CRF08_BC subtype, the frequencies of V77I and K277R in CRF07_BC subtype, and the frequencies of M36I and L89M in CRF01_AE subtype were higher than those in the other two groups, and the differences were all statistically significant (all P<0.017). Conclusions:The polymorphic loci resulting from HIV-1 HAART failure show different distribution patterns across various degrees of drug resistance, treatment regimens and HIV-1 subtypes.These loci demonstrate both specific and shared characteristics. It is necessary to enhance the surveillance of select polymorphic loci.

Objective:To analyze the variation characteristics of gag gene sequence of human immunodeficiency virus type 1 (HIV-1) epidemic strains in four major acquired immunodeficiency syndrome (AIDS) endemic areas in Yunnan Province. Methods:A total of 480 human immunodeficiency virus (HIV)/AIDS patients from designated antiviral treatment institutions in Kunming City, Dehong Dai and Jingpo Autonomous Prefecture, Hani-Yi Autonomous Prefecture of Honghe and Lincang City in Yunnan Province from January 2019 to December 2020 were randomly selected. The epidemiological information of the patients was collected. The plasma samples were collected and sent to the Yunnan Provincial Infectious Diseases Hospital for analysis. HIV-1 gag gene was amplified by nested polymerase chain reaction for genotyping.The phylogenetic tree was constructed by MEGA 6.06, and the characteristic amino acids were analyzed by VESPA online analysis tool. The gene distances of gag and matrix protein p17, capsid protein p24, nucleocapsid protein p7 and p6 protein of gag protein were calculated by Distance program. The ratio of synonymous mutation frequnency and non-synonymous mutation frequnency (Ks/Ka) was analyzed by SNAP program. Statistical comparison among multiple groups was performed using analysis of variance. Results:The gag gene sequences were successfully obtained from 404 patients.Most of these patients were men (250 cases, 61.9%), 59.7%(241/404) of patients were aged 40 to 59 years, and the main transmission route was heterosexual transmission (61.4%, 248/404). The main epidemic subtypes of HIV-1 were circulating recombinant form(CRF)08_BC (38.4%, 155/404), CRF01_AE (18.3%, 74/404), unique recombinant form (URF) (12.9%, 52/404), CRF07_BC(9.7%, 39/404), C subtype (8.4%, 34/404), other subtypes (6.9%, 28/404) and B subtype (5.4%, 22/404). Two main spreading clusters were formed by CRF08_BC in the phylogenetic tree, and there were significant differences in the distribution of characteristic amino acids of eight loci between the two spreading clusters, including No.79, 93, 121, 122, 151, 363, 395 and 396. The gag gene distances of CRF01_ AE, CRF07_ BC and CRF08_ BC were 0.090±0.004, 0.088±0.004 and 0.078±0.002, respectively, and the difference was statistically significant ( F=118.33, P<0.001). The Ks/Ka values of gag of the three subtypes were 4.003±1.309, 4.141±0.860 and 4.514±1.215, respectively, and the difference was statistically significant ( F=1.35, P<0.001). The Ks/Ka values of p17 were 2.590±0.186, 2.831±0.496 and 2.936±0.475, respectively, and the difference was statistically significant ( F=1.59, P<0.001). The Ks/Ka values of p24 were 12.579±1.116, 10.185±0.494 and 8.522±0.595, respectively, and the difference was statistically significant ( F=1.61, P<0.001). The Ks/Ka values of p7 were 10.850±0.711, 9.717±0.932 and 8.522±0.026, respectively, and the difference was statistically significant ( F=4.24, P<0.001). The Ks/Ka values of p6 were 3.122±0.134, 3.040±1.498 and 4.841±0.353, respectively, and the difference was statistically significant ( F=10.68, P<0.001). Conclusions:CRF08_ BC is the main subtype in these four areas in Yunnan Province.The different clusters of CRF08_BC result in a different pattern of amino acid composition.The variation degree of different subtype gag gene is different in each section. Surveillance of HIV variant strains should be strengthened to control the epidemics of HIV.

Objective:To understand the contents of edible salt iodine and urinary iodine of children and pregnant women in Yunnan Province, and to evaluate the iodine nutrition status, so as to provide a basis for scientific prevention and treatment of iodine deficiency disorders (IDD).Methods:From November to December 2020, one county (city) was selected from each prefecture (city), two townships (towns and streets) were selected from each county (city) and two villages (neighborhood committees) were selected from each township (town and street) from each of the 16 prefectures (cities) in Yunnan Province as the investigation sites. A total of 20 non-boarding children (male and female balanced) aged 8 - 10 years old were selected from each primary school in each village (neighborhood committee) to collect salt and urine samples. A total of 80 children were investigated in each county (city). A total of 20 pregnant women were selected from each township (town and street) to collect salt and urine samples. A total of 40 pregnant women were investigated in each county (city). All salt samples and urine samples were tested for iodine contents.Results:A total of 2 009 salt samples and 2 041 urine samples (1 375 for children, 666 for pregnant women) were collected from children aged 8 - 10 years old and pregnant women in 16 counties (cities) of Yunnan Province. Among them, the median salt iodine was 26.0 mg/kg, the coverage rate of iodized salt was 100.0% (2 009/2 009), the qualified rate of iodized salt was 98.7% (1 982/2 009), and the consumption rate of qualified iodized salt was 98.7% (1 982/2 009). The difference of salt iodine content in key populations in different counties (cities) was statistically significant ( H = 258.98, P < 0.01). The median urinary iodine of children aged 8 - 10 years old was 188.5 μg/L. There was statistically significant difference in urinary iodine content among children of different ages ( H = 29.45, P < 0.01), but there was no statistically significant difference in urinary iodine content among children of different genders ( H = 1.43, P > 0.05). In addition, the median urinary iodine of pregnant women was 141.9 μg/L, 52.1% (347/666) was < 150 μg/L. There was statistically significant difference in urinary iodine content of pregnant women in different counties (cities, H = 88.32, P < 0.01). Conclusions:The qualified rate of iodized salt, the coverage rate of iodized salt and the consumption rate of qualified iodized salt in key populations of Yunnan Province are more than 90%, and the iodized salt supply is good. Iodine nutrition of children aged 8 - 10 years old is at an appropriate level (100 - 199 μg/L); iodine nutrition of pregnant women is in an state of iodine deficiency ( < 150 μg/L). It is suggested to strengthen IDD monitoring and health education among key populations, improve residents' awareness of disease prevention, and make scientific iodine supplementation.

Objective: To learn the genotypic drug resistance of men who have sex with men ( MSM ) with HIV who failed in antiviral therapy in Yunnan Province, in order to provide basis for improving the effect of antiviral therapy. Methods: The patients who were infected with HIV-1, homosexual transmitted and failed in antiviral therapy in Yunnan Province from 2014 to 2019 were recruited. Their plasma samples were tested by reverse transcription nested polymerase chain reaction ( RT-nPCR ) , the fragments were spliced using ContigExpress, and the resistance to 8 protease inhibitors ( PIs ) , 7 nucleoside reverse transcriptase inhibitors ( NRTIs ) and 5 non-nucleoside reverse transcriptase inhibitors ( NNRTIs ) were obtained from the HIV drug resistance data website of Stanford University. Results: A total of 205 HIV/AIDS cases were included, 169 positive plasma samples were amplified, 112 cases were drug resistant, and the rate of drug resistance was 66.27%. The patients who were aged 30-49 years ( 76.09% ) , had genotype of CRF01_AE ( 76.34% ) or treated by AZT+3TC+NVP ( 77.08% ) had higher resistance rate. The resistance rates of NNRTIs, NRTIs and PIs were 62.72%, 49.70% and 2.96%, respectively; the resistance rates of NVP and EFV in NNRTIs were 62.72% and 61.54%. The main mutation site associated with NNRTIs was K103, accounting for 21.89% ( 37 cases ) ; the main mutation site associated with NRTIs was M184, accounting for 39.64% ( 67 cases ) ; the main mutation sites associated with PIs were M46L/K, accounting for 2.96% ( 5 cases ) , resulting in high resistance to NFV. Conclusions The drug resistance rate of HIV-infected MSM with failure of antiviral therapy in Yunnan Province is relatively high, with CRF01_AE as the main gene subtype of drug resistance. The drug resistance rate of NNRTIs is relatively high, especially NVP and EFV.

Objective To investigate the influence factors of mortality among human immunodeficiency virus (HIV)-infected children under highly active antiretroviral therapy (HAART).Methods Retrospective cohort study of 652 children initiated HAART from 2005 to 2014 was conducted,and enrolled patients were followed-up until December,2015.Survival data was analyzed using Kaplan-Meier method and Cox regression model was used to identify independent predictors of mortality among these children on HARRT.Chi-square test and Fisher's exact test were used for comparison between groups.Results Overall,26 of the children died over a follow-up period of 3 116.24 child-years,with a mortality rate of 0.83 per 100 child-years.Twelve (46％)of deaths occurred during the first six months after starting HAART.Cox regression analysis of variables showed that the World Health Organization (WHO) clinical stages Ⅲ/Ⅳ (hazard rate [HR] =10.717,95％confidence interal [95％ CI]:4.189-4.749,P =0.000),baseline hemoglobin ＜ 80 g/L (HR =14.768,95 ％ CI:5.721-38.125,P =0.000),tuberculosis co-infection (HR =4.794,95％ CI:2.105-10.918,P =0.000),baseline CD4+T lymphocyte ＜ 50 cells/μL (HR =4.219,95％ CI:1.524-11.680,P =0.006),weight-for-age z-score ＜-2 (HR =2.983,95 ％ CI:1.094-8.135,P =0.033) were independently associated with death,whereas the age ＜ 7 year-old at HAART initiation was protectire (HR =0.293,95％ CI:0.126-0.684,P =0.005).Conclusions The mortality of children receiving HAART is strongly associated with WHO stages Ⅲ/Ⅳ,hemoglobin ＜ 80 g/L,weight-for-age z-score ＜-2,tuberculosis co-infection and older age at treatment.

Objective To compare the differences of CD4 +T lymphocyte (CD4) counts between patients aged 18 and over,to explore the effect of age on treatment,36 months after having received the China National Free AIDS Antiretroviral Treatment on HIV/AIDS.Methods Through the National ART Information Ssystem,we selected those HIV/AIDS patients who initiated the ART 36 months after the ART,between January 1,2010 and December 31,2012 in Guangzhou,Liuzhou and Kunming.Patients were divided into age groups as 18-49,50-59 and 60 or over year olds,at the baseline of treatment.Under different levels of baseline CD4 counts,we chose the baseline and different time-point of CD4 counts as dependent variables,applied mixed linear model to analyze the effects of age,viral suppression,gender,baseline CD4/CDs ratio and initial treatment regimen.Results A total of 5 331 HIV/AIDS patients were recruited.No differences were found on age group ratios between different levels of baseline CD4 counts.At the level of baseline CD4＜200 cells/μl,both the 50-59 and 60 or above years old groups had lower CD4 counts than the 18-49 year-old group,within 36 months after the initiation of ART.However,at the baseline CD4 level of 200-350 cells/μl,no signiftcant differences on CD4 counts between the 50-59 year-old and 18-49 year-old groups were noticed.CD4 counts seemed lower in the 60 and above year-old group than in the 18-49 year-old group.Conclusion Age might serve as an influencing factor on CD4 counts within 36 months after the initiation of ART,suggesting that earlier initiation of ART might be of help to the recovery of immune function in the 50-59 year-old group.

Objective To investigate the subtypes distribution of human immunodeficiency virus (HIV)-1 epidemic strains and the characteristics of amino acid variation in different areas of Yunnan Province.Methods Totally 800 HIV/AIDS plasma specimens and epidemiological information were collected between 2012 and 2015 from 14 areas of Yunnan.Viral RNA was extracted and amplified using RT polymerase chain reaction (PCR).4.5 kb 5'halves fragments were obtained and directly sequenced.Subtypes of strains were identified by Genotyping,MEGA 6.06 and BLAST.Grouping was analyzed by location and subtype.Entropy software was used to analyze the difference of amino acid sequences between different groups according to the sampling location and subtypes to analyze the regional distribution and genetic variation of HIV-1 subtypes in Yunnan Province.Results Of the 800 plasma specimens,a total of 446 genomic sequences from 12 areas were successfully amplified and sequenced.After genotypes were identified,the subtypes of HIV-1 strains prevalent in Yunnan were CRF08_BC (58.3％),CRF01_AE (19.3％),CRF07_BC (11.6％),unknown recombinant forms (7.1％),B(B') (1.7％) and C (1.3％).The geographical distribution in Yunnan was analyzed.The CRF01_AE predominated in Dehong,Xishuangbarma and Wenshan.The CRF08_BC predominated in Lincang,Honghe and Puer (more than 70.0％) and CRF08_BC was prevalent in the other areas.But CRF07_BC in Kunming,Yuxi and Dali accounted for more than 20 ％.The constitutions of amino acid of three majors CRF08_BC,CRF01 _AE and CRF07_BC were different on 17,14 and 18 amino acid sites with statistical differences in the eastern and western regions of Yunnan Province (均P ＜ 0.05).Conclusions HIV-1 strains transmit and vary genetically in the province widely.The amino acid mutation sites of eastern and western strains are different.This difference represents that the same subtype strains in different geographical distribution vary on different genetic background and are selected by immune responses.The epidemic trends need to be closely monitored.

OBJECTIVETo investigate the distribution patterns of mosquito and mosquito-borne viruses in Dehong prefecture, Yunnan province, China.

METHODSMosquito samples were collected using the mosquito traps from five counties of Dehong prefecture on July, 2007 and 2010. Mosquito were cell cultured for viral isolation, and positive isolates were identified using RT-PCR and sequence analysis.

RESULTSA total of 43 634 mosquito comprised of 29 species representing six genera were collected. Culex tritaeniorhynchus and Anopheles sinensis comprised 78.69% and 14.77% of the total. Six strains of viruses were isolated from the mosquito pools. RT-PCR and phylogenetic analysis revealed three strains from Cx. tritaeniorhynchus, identified as genotype I Japanese encephalitis virus (JEV). One strain was identified from Cx. tritaeniorhynchus, as Getah virus (GETV). Two strains isolated from Cx. tritaeniorhynchus and Anopheles vagus were identified as Culex pipiens pallens Densovirus (CppDNV).

CONCLUSIONCx. tritaeniorhynchus had been the major species of mosquito and mainly transmitting vector of mosquito-borne viruses in Dehong prefecture. Genotype I JEV, GETV and CppDNV were the vectors causing transmission of mosquito-borne diseases in this area. Data from phylogenetic analysis showed that these newly discovered isolates seemed to have had close relationship with those viruses previously circulating in Yunnan and other provinces of China.

Alphavirus ; isolation & purification ; Animals ; Arboviruses ; classification ; isolation & purification ; China ; Culicidae ; virology ; Disease Vectors ; classification ; Encephalitis Virus, Japanese ; isolation & purification