Objective To establish a duplex fluorescence PCR for detection of HIV proviral DNA and to evaluate its application for early diagnosis of HIV infection in infants .Methods A duplex fluores-cence PCR system was set up based on TaqMan technology for detection of human ribonuclease P ( RNase P) gene and long terminal repeat ( LTR) region of HIV.A recombinant plasmid containing the targeted gene fragment , pTG19-T, was constructed by TA cloning technique and used as the template for evaluation of sen -sitivity of the assay .Blood samples from 11 healthy individuals and 98 HIV-infected patients were collected and detected to validate the assay specificity .The assay of duplex fluorescence PCR was then carried out to detect 96 infant blood samples collected from several maternal and child health hospitals in Zhejiang province from January 2011 to September 2012 for early diagnosis of HIV infection .The results were compared with those by using the Roche HIV DNA qualitative detection kit .Results The established duplex fluorescence PCR could specifically detect HIV proviral DNA with a specificity of 100%and a detection sensitivity of 100 cps per reaction .The coincidence rate between the established assay and the Roche HIV DNA qualitative de -tection kit was 100%in the detection of 96 blood samples .Conclusion The duplex fluorescence PCR as-say showed advantages of cost-effectiveness , convenience , good specificity and accuracy with high sensitivi-ty.It could be used for early diagnosis of HIV infection in infants and also as a general technical platform for the detection of HIV proviral DNA .

Helicobacter pylori (Hp) is a microaerophilic Gram-negative bacterium that colonizes the gastric mucosa. After colonization, many proteins expressed by Hp, such as Helicobacter pylori urease (HPU), neutrophil-activating protein (NAP), outer membrane protein (OMP), cytotoxin-associated gene A (CagA) and vacuolating cytotoxin A (VacA), may serve as antigens to mediate neutrophil chemotaxis. Thereafter, neutrophils infiltrate in gastric mucosa and play roles in mucosal immunity via secreting cytokines and releasing reactive oxygen species (ROS) and myeloperoxidase (MPO). This article reviewed the advances in study on the role of neutrophil infiltration in Hp infection-related diseases.

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.

Objective To explore the nursing effect of home care in patients with minimally invasive total hip arthroplasty (THA) in patients with minimally invasive surgery,and to provide comprehensive and seamless care for patients.Methods A total of 79 patients with THA were divided into observation group (38 cases)and control group(41 cases).The observation group received home care,including health education,telephone follow-up,home visits and other nursing measures.Control group followed the routine nursing education.Activity of Daily Living Scale(ADL),Functional Independence Measure(FIM) and Harris scores were compared between the two groups of patients when discharged and three months after discharge.Results In the observation group and the control group,the ADL,FIM and Harris scores were gradually increased at 3 months after discharge which had no difference when discharge (P ＞ 0.05).The ADL,FIM and Harris scores of the observation group were significantly higher than that in the control group at 3 months after discharge (P ＜ 0.01).Conclusions Implementation of home care can improve patients' ability of daily life,the whole hip joint function recovery and hip function independence,thus it makes their quality of life better,achieving the goal of comprehensive rehabilitation.

Objective:To compare the performance of four commercial kits for anti-HIV confirmatory assay and to provide a reference for clinical application.Methods:Two western blot (WB) kits and two recombinant immunoblot assay/line immune assay kits were used to detect 185 samples in parallel, McNermar test was used to analyze the consistency between the testing results and HIV infection status. Compared to the testing result of the most used MP company’s WB kit, the odds ratio ( OR) was calculated. Results:The consistency of the testing results with the status of HIV infection by the four kits was 83.24%-92.43%, and the Kappa value was 0.558-0.831.The OR value of IMT company’s WB kit obtained from 57 HIV-uninfected samples was 0.932(95% CI: 0.446-1.946, P=0.851), and the OR values of WT company’s recombinant immunoblot assay (RIBA) and MK company’s line immune assay (LIA) were 2.348(95% CI: 1.069-5.158, P=0.034) and 23.064 (95% CI: 5.125-103.789, P<0.001), respectively. The OR value of LIA obtained from 128 HIV-infected samples was 0.153 (95% CI: 0.034-0.700, P=0.016). LIA showed fewest reactions by testing 95 HIV positive screening samples, which was significantly different from any other kit ( P< 0.05). There was no significant difference among the other three kits. In the case of non-specific reactions, the numbers of bands were all ≤4, and the proportion of light-colored bands accounted for 52.1% (37/71). The main bands were gp160(81.3%), p24(71.9%), and p17(28.1%). Conclusions:There was no significant difference between the two WB kits. The RIBA/LIA detection kits could reduce the generation of indeterminate result, but the LIA kit has the risk of false negative for weakly reactive samples.

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 H1V/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.

Objective? To expLore the effects of Home FaLL Hazards Assessment (HFHA) on the incidence rate of home faLLs and recovery of hip function in patients receiving totaL hip arthropLasty (THA). Methods? TotaLLy 423 patients who received uniLateraL THA for the first time in the Department of Orthopedics, Nanjing Drum Tower HospitaL admitted from JuLy 2015 to JuLy 2017 were seLected by convenient sampLing and divided into the controL group (n=203) and the treatment group (n=220). Patients in the controL group received conventionaL postoperative nursing care and discharge guidance, whiLe the home faLL hazards of patients in the treatment group were evaLuated by HFHA in addition to conventionaL postoperative nursing care and discharge guidance. Nursing care was provided to them based on the assessment resuLts. The joint function score at discharge and 3 months after discharge, the incidence rate of faLLs over the past one year and the severity of injury caused by faLLs were compared between the two groups. ResuLts? There was no statisticaL difference in Harris index and BartheL index scores between the two groups at discharge (P＞ 0.05). Harris index and BartheL index scores 3 months after discharge were statisticaL differences in both groups (P＜0.05). 55 faLLs occurred over the past one year in the controL group, 7 of them with moderate or above injuries, whiLe 35 faLLs occurred over the past one year in the treatment group, 1 of them with moderate or above injuries. There were statisticaL differences in the incidence rate of faLLs and injury degrees between the two groups (P＜0.05). ConcLusions? HFHA, when used to assess the home faLL hazards, enabLes THA patients to know and note the high-risk hazards contributing to faLLs in their environment of rehabiLitation and effectiveLy reduces the patients home faLLs by improving the home environment.

Objective:To investigate the factors associated with delay in anticoagulant therapy in patients with cerebral venous sinus thrombosis (CVST) and its effect on outcome.Methods:Patients with CVST admitted to Changhai Hospital, Naval Medical University from January 2010 to August 2021 were retrospectively enrolled. Patients were divided into early anticoagulation group and late anticoagulation group by the median time interval from first symptom to initiation of anticoagulation. The modified Rankin Scale was used for outcome assessment at 90 d after onset. 0-2 scores were defined as good outcome and 3-6 were defined as poor outcome. Demographic and clinical data were compared for the early versus late anticoagulation group and for the good versus poor outcome groups. Multivariable logistic regression was used to identify independent influencing factors of delay in anticoagulation and the correlation of delay in anticoagulation with poor outcome. Results:A total of 131 patients were included, their age was 40.07±15.11 years old, and 68 (51.91%) were male. Of these, 65 patients (49.62%) were in the early anticoagulation group and 14 (10.69%) were in the poor outcome group. Compared with the late anticoagulation group, the early anticoagulation group had a significantly higher proportion of patients with seizures and brain parenchymal damage as well as higher D-dimer levels on admission, while the proportion of patients with visual impairment/papilloedema was significantly lower (all P<0.05). Compared with the good outcome group, the poor outcome group had significantly higher proportions of patients with seizures, dyskinesia, impaired consciousness, low Glasgow Coma Scale score, and brain parenchymal damage as well as higher D-dimer, total cholesterol and low density lipoprotein cholesterol levels, sites of thrombus involvement were more common in the superior sagittal and straight sinuses, and significantly lower proportions of patients with headache and lower albumin levels on admission (all P<0.05). Multivariate logistic regression analysis showed that visual impairment/papilloedema (odds ratio [ OR] 0.119, 95% confidence interval [ CI] 0.030-0.473; P=0.002) and brain parenchymal damage ( OR 1.341, 95% CI 1.042-1.727; P=0.023) were independently associated with a delay in anticoagulation treatment, and a delay in anticoagulation treatment ( OR 6.102, 95% CI 1.185-30.504; P=0.030) and D-dimer level on admission ( OR 1.299, 95% CI 1.141-1.480; P<0.001) were the independent predictors of poor outcome in patients with CVST. Conclusions:Visual impairment/papilloedema and absence of brain parenchymal damage on cranial imaging are the independent risk factors for delay in anticoagulation in patients with CVST. The delay in anticoagulation is strongly associated with the poor outcome in patients with CVST.

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) ( χ2=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.

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.