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ObjectiveThis study aimed to investigate the HIV genotypic subtypes and molecular transmission clusters among men who have sex with men （MSM） with newly reported HIV infections in Dehong Dai and Jingpo Autonomous Prefecture （Dehong Prefecture）， Yunnan Province， China， between 2010 and 2019. The study aimed to identify potential high-risk transmitters and provide reference data for screening， management， and intervention of infection sources. MethodsPlasma samples from newly reported HIV-positive MSM individuals in Dehong Prefecture between 2010 and 2019 were collected. The viral pol gene fragments were amplified， sequenced， and genotyped. Genetic distances （GD） between pairwise sequences were analyzed and calculated. MEGA 7.0 and Gephi were used for phylogenetic and molecular transmission network analysis. ResultsA total of 159 newly reported HIV infections among MSM were included in the study， with successful genotyping of 100 cases. Nine HIV-1 subtypes were identified， with the most prevalent being CRF01_AE subtype （52%）， followed by CRF07_BC subtype （31%）， CRF55_01B subtype （10%）， and others （7%）. Cluster analysis revealed a total network access rate of 67%， forming three transmission clusters. CRF01_AE subtype formed two transmission clusters with 38 and 3 infected individuals， while CRF07_BC subtypes formed one transmission cluster with 26 infected individuals. The transmission network within the CRF01_AE clusters exhibited a more complex relationship. Significant differences in educational level were observed between the two main transmission clusters. ConclusionThe predominant HIV subtypes among newly reported MSM cases in Dehong Prefecture between 2010 and 2019 were CRF01_AE and CRF07_BC. Significant cultural differences are observed between the main transmission clusters. Continued monitoring of genotypic subtypes and targeted interventions within transmission clusters are warranted.
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Objective:To investigate the distributions of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) genotypes among newly reported HIV/HCV co-infected Burmese patients in Dehong Dai and Jingpo Autonomous Prefecture, Yunnan Province from 2016 to 2019.Methods:A total of 1 289 newly reported HIV/HCV co-infected Burmese patients in Dehong Dai and Jingpo Autonomous Prefecture were collected through the National Acquired Immunodeficiency Syndrome Comprehensive Prevention and Control Data Information System From January 2016 to December 2019. Among them, 996 subjects with a plasma volume of ≥200 μL were selected to perform HIV and HCV genotyping. The HIV pol gene, the HCV core protein-binding envelope protein ( CE1) gene and non-structural protein 5B ( NS5 B) gene were amplified using the nested polymerase chain reaction.The phylogenetic tree was constructed by MEGA 7.0 software to classify the genotypes. Chi-square test was used for statistical analysis. Trend chi-square test was used to analyze the trend of HIV and HCV genotypes. Results:Among the 996 cases with HIV/HCV co-infection, HIV and HCV sequences from a total of 554 subjects (55.6%, 554/996) were successfully obtained, and the genotypes of HIV and HCV were diverse. HIV genotype C (40.3%, 223/554) and BC recombinant (33.6%, 186/554) were the most prevalent, followed by genotype B (6.5%, 36/554) and circulating recombinant form (CRF)01_AE (3.6%, 20/554). HCV genotype 3b was the most prevalent (31.2%, 173/554), followed by genotype 6u (19.5%, 108/554), 1a (17.5%, 97/554), 6n (11.4%, 63/554), 3a (8.7%, 48/554) and 6xg (6.3%, 35/554). The prevalence of HIV genotype C showed a downward trend ( χtrend2=7.23, P<0.001), while the prevalence of BC recombinant showed an upward trend ( χtrend2=5.97, P<0.001), and the proportion of BC recombinant was higher than genotype C in 2019 (54.9%(101/184) vs 21.7%(40/184)). However, there were no statistically significant differences in the proportions of genotype 3b, 6u and 1a from 2016 to 2019 ( χtrend2=1.43, 1.79 and 0.39, respectively, P=0.152, 0.074 and 0.695, respectively). The HIV genotype distribution among patients with different ethnic groups were significantly different ( χ2=22.06, P=0.037). Conclusions:The diversity of HIV and HCV genotypes is high and complex among HIV/HCV co-infected Burmese patients in Dehong Dai and Jingpo Autonomous Prefecture. BC recombinant shows a trend of becoming the predominant HIV genotype among these co-infected patients. Therefore, surveillance of the prevalence of HCV and HIV genotypes in Burmese population needs to be further strengthened.
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Objective To investigate the role of LncRNAs-MEG3 in the carcinogenesis and progression of nasopharyngeal carcinoma (NPC) and the possible molecular mechanism. Methods qRT-PCR was used to detect the content of MEG3 and miR-543 in NPC cells. Luciferase reporter method was used to study the relation between MEG3 and miR-543, and the changes of cell proliferation and apoptosis induced by MEG3 or KLF4 were analyzed. Western blot was used to detect the expression of KLF4, Bcl-2 and Bax proteins. Results Compared with the control group, the expression of miR-543 in NPC cell line was significantly increased (P < 0.05), while the expression of MEG3 was decreased (P < 0.05). Luciferase report and Western blot showed that MEG3 could regulate the expression of KLF4 by adsorbing miR-543 to inhibit cell proliferation, promote cell apoptosis and affect the expression levels of Bcl-2 and Bax proteins. Conclusion LncRNA-MEG3 could regulate the expression of KLF4 by adsorbing miR-543 and then plays a role in inhibiting the occurrence and development of NPC. It may be a new biomarker for NPC targeted therapy.
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Objective To understand the characteristics on major strain subtypes of hepatitis C virus among HIV/HCV co-infected patients,so as to explore the molecular transmission clusters and related risk factors of HCV strains.Methods A total of 336 newly reported HIV-infected patients were diagnosed as HIV/HCV co-infection in Dehong Dai and Jingpo autonomous prefecture (Dehong)in 2016.We used Nested PCR to amplify CE 1 and NS5B genes among 318 samples with plasma levels above 200 μl,before using the combining phylogenetic tree and constructing molecular propagation network method to analyze the related data.Results A total of 267 HIV/HCV co-infection patients who had met the HCV genotyping requirements were screened the gene subtypes were diversified.Among these genotypes,proportions of 3b,6n,6u,1a,3a and other subtypes appeared as 32.6％ (87/267),18.4％ (49/267),15.7％(42/267),13.1％(35/267),11.2％(30/267) and 9.0％(24/267) respectively.Molecular transmission network of five major HCV genotypes was constructed with a clustering rate of 39.1％ (95/243).The clustering rate of subtype la was the highest,as 71.4％ (25/35).Results from the multivariate logistic regression showed that ethnic minorities other than the Yi and Jingpo (vs.the Han,OR=0.17,95％ CI:0.04-0.71),the married spouses (vs.the unmarried,OR=0.42,95％ CI:0.18-0.94),the 6n and 3a subtype (vs.the 3b subtype,OR=0.34,95％CI:0.12-0.95;OR=0.22,95％CI:0.05-0.93) were more difficult to form transmission clusters.However,the 6u and 1a subtype (vs.the 3b subtype,OR=3.10,95％CI:1.21-7.94;OR=4.00,95％CI:1.32-12.11) seemed more likely to form the transmission clusters.Conclusion Ethnicity,marital status and genetic subtypes were factors significantly associated with the formation of transmission clusters related to the major HCV gene subtypes among newly reported HIV/HCV co-infection in Dehong.
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Objective To understand the characteristics on major strain subtypes of hepatitis C virus among HIV/HCV co-infected patients,so as to explore the molecular transmission clusters and related risk factors of HCV strains.Methods A total of 336 newly reported HIV-infected patients were diagnosed as HIV/HCV co-infection in Dehong Dai and Jingpo autonomous prefecture (Dehong)in 2016.We used Nested PCR to amplify CE 1 and NS5B genes among 318 samples with plasma levels above 200 μl,before using the combining phylogenetic tree and constructing molecular propagation network method to analyze the related data.Results A total of 267 HIV/HCV co-infection patients who had met the HCV genotyping requirements were screened the gene subtypes were diversified.Among these genotypes,proportions of 3b,6n,6u,1a,3a and other subtypes appeared as 32.6％ (87/267),18.4％ (49/267),15.7％(42/267),13.1％(35/267),11.2％(30/267) and 9.0％(24/267) respectively.Molecular transmission network of five major HCV genotypes was constructed with a clustering rate of 39.1％ (95/243).The clustering rate of subtype la was the highest,as 71.4％ (25/35).Results from the multivariate logistic regression showed that ethnic minorities other than the Yi and Jingpo (vs.the Han,OR=0.17,95％ CI:0.04-0.71),the married spouses (vs.the unmarried,OR=0.42,95％ CI:0.18-0.94),the 6n and 3a subtype (vs.the 3b subtype,OR=0.34,95％CI:0.12-0.95;OR=0.22,95％CI:0.05-0.93) were more difficult to form transmission clusters.However,the 6u and 1a subtype (vs.the 3b subtype,OR=3.10,95％CI:1.21-7.94;OR=4.00,95％CI:1.32-12.11) seemed more likely to form the transmission clusters.Conclusion Ethnicity,marital status and genetic subtypes were factors significantly associated with the formation of transmission clusters related to the major HCV gene subtypes among newly reported HIV/HCV co-infection in Dehong.
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Objective To explore distribution of HIV gene subtypes among newly reported HIV/AIDS cases from China and Myanmar in Dehong Dai and Jingpo prefecture of Yunnan province in 2016.Methods We conducted DNA extractions from newly reported HIV/AIDS cases in 2016.The gag,env and pol genes were amplified by using reverse transcription-PCR (RT-PCR) and sequenced to identify HIV subtypes.Results A total of 1 112 newly diagnosed HIV cases were reported in Dehong in 2016,and the HIV subtypes were identified for 860 cases.Subtype C was predominant (33.6％),followed by unique recombinant forms (URFs) (28.4％),CRF01_AE (18.6％) and so on.URFs include four recombination,among which the recombination of CRF01 AE and C subtype were predominant.The HIV subtype distribution was associated with nationality and transmission route in HIV/AIDS cases from Myanmar.Conclusions The gene subtypes of C,URFs and CRF01_AE were mainly distributed;distribution of URFs remained complex and diverse among newly reported HIV/AIDS cases in Dehong in 2016.
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Objective To explore distribution of HIV gene subtypes among newly reported HIV/AIDS cases from China and Myanmar in Dehong Dai and Jingpo prefecture of Yunnan province in 2016.Methods We conducted DNA extractions from newly reported HIV/AIDS cases in 2016.The gag,env and pol genes were amplified by using reverse transcription-PCR (RT-PCR) and sequenced to identify HIV subtypes.Results A total of 1 112 newly diagnosed HIV cases were reported in Dehong in 2016,and the HIV subtypes were identified for 860 cases.Subtype C was predominant (33.6％),followed by unique recombinant forms (URFs) (28.4％),CRF01_AE (18.6％) and so on.URFs include four recombination,among which the recombination of CRF01 AE and C subtype were predominant.The HIV subtype distribution was associated with nationality and transmission route in HIV/AIDS cases from Myanmar.Conclusions The gene subtypes of C,URFs and CRF01_AE were mainly distributed;distribution of URFs remained complex and diverse among newly reported HIV/AIDS cases in Dehong in 2016.
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An efficient method for the analysis of multiclass plant growth regulators and pesticide (imidacloprid, acetamiprid) residues in tea was developed based on ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The samples were extracted with acetonitrile/formic acid (99∶1, V/V) solution, cleaned up with four sorbents including C18, strong anion exchanger (SAX), primary secondary amine (PSA) and anhydrous MgSO4. The compounds were separated on a HSS T3 column under positive/negative electrospray ionization mode, detected by scheduled multiple reaction monitoring (SMRM), and quantified by matrix-matched external standard curves. All pesticide residues showed good linearity in the concentration range of 1-200 μg/L (6-benzylaminopurine, paclobutrazol, uniconazole, forchlorfenuron, mepiquat chloride, imidacloprid, acetamiprid) or 5-1000 μg/L (2,4-dichlor-ophenoxyacetic acid, 4-chlorophenoxyacetic acid, indole-3-acetic acid, gibberellic acid, 1-naphthaleneacetic acid, indole-3-butyric acid) , with correlation coefficient (R2≥0.99). Limits of detection (LOD, S/N=3) and limits of quantitation (LOQ, S/N=10) were 0.18-9.68 μg/kg and 0.61-32.26 μg/kg, respectively. In addition, the spiked recoveries of tea samples were 73.1％-108.9％, and RSDs were 0.6％-8.0％. This method was applied to commercial samples, and all the detections were confirmed by acquiring transitions for each pesticide in the samples.
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Comprehensive geriatric assessment (CGA) is widely used for creating tailored individual care plans for seniors who usually are suffering from multiple diseases,frailty,disability and cognition decline.Skilled communicators are of importance for high-quality clinical data collection.Communication skills used in CGA are summarized in this article by reviewing related references and combining practical experiences in the process of applying international resident assessment instruments (interRAI).
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An efficient technique for the determination of multiclass pesticide residues ( glyphosate, aminomethyl phosphonic acid ( AMPA) and glufosinate) in four kinds of tea ( green tea, black tea, Oolong tea and Puer tea) was developed based on ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Samples were extracted with 0. 05 mol/L NaOH solution, regulated pH value with HCl, followed by purification by Oasis HLB column, and then precolumn-derived with 9-fluorenylmethyl chloroformate ( FMOC-CL ) in borate buffer. All pesticide residues studied showed good linearity with correlation coefficient ( R2 ) greater than 0. 99 in the concentration range of 5-1000 μg/L. Limits of quantitation (LOQs) of three pesticide compounds ranged from 0. 03 to 0. 08 mg/kg (S/N=10). Tea samples spiked with 0 . 1 , 0 . 4 and 4 mg/kg showed the recovery ranging from 72 . 1% to 109 . 9%, and RSDs from 0. 5% to 9. 8% (n=6). In addition, the validated method was applied to commercial samples, and all the detections were confirmed by acquiring transitions for each pesticide in samples.
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Objective To optimize the preparative procedure for stachydrine in Fructus Leonuri. Methods The preparation was screened by orthogonal experiment, and a mathematical model of relationship of extraction time, methanol concentration, and solid-liquid ratio with the content of stachydrine hydrochloride was established by using back-propagation (BP) neural network. And the process parameters were optimized with genetic algorithm (GA) . Results The optimum process parameters were as follows: extraction with 69% of methanol concentration and with solid-liquid ratio being 11 times for 62 min. The content of stachydrine obtained by BP neural network modeling and GA was higher than that achieved by orthogonal experiment. Conclusion The optimum preparative procedure could be achieved by combining BP modeling with GA. The model developed in this study was proved to be predictable and feasible for the optimization of process parameters of multi-dimension nonlinear system.
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Objective To study the efficacy and safety of CO2 laser microsurgery for early glottis carcinoma.Methods The data of 96 cases of early glottic laryngeal carcinoma treated with CO2 laser microsurgery were retrospectively analyzed.Surgical specimens were conventionally embedded with paraffin and serially sectioned.Sections were stained by using hematoxylin-eosin for pathologic examination.The local control rate was observed after operation.Results Among the 96 specimens,88 surgical margins were negative and 8 were positive.10 patients (10.4 ％) recurrenced followed up for 3 years.The recurrence rates of Tis,T1a,T1b and T2 were 0 (0/10),4.1％ (2/48),25.0 ％ (4/16) and 18.1％ (4/22),respectively,with significant differences among groups (X2 =6.105,P ＜ 0.05).All of 8 cases with positive margin and 2 cases with negative margin were recurrened.The recurrent rate of patients with involvement of the anterior commissure was 30.0 ％ (6/20),which was higher than that of patients with no tumor invasion [5.3 ％ (4/76)] (X2 =9.624,P ＜ 0.01).Conclusions The safe edges can be obtained by CO2 laser microsurgery for early stage of glottis carcinoma,which have advantages in local control rate,vocal function of the larynx and curative effect.