Objective:To provide data support for acquired immunodeficiency syndrome (AIDS) clinical treatment by evaluating the antiviral therapy effect of integrase inhibitors (INIs) and analyzing the drug resistant mutations.Methods:Patients who exposed to INIs more than 1 year were analyzed retrospectively. Virus suppression failure subjects(viral load ≥1 000 copies/ml) were sequenced for drug resistance analysis.Results:Of 408 patients, 12 cases(2.9%) were virologic failure, 8 cases had drug resistance mutations. The proportion of drug resistance was 2.0%(8/408). Among them, only 1 case(0.2%, 1/408) was resistant to INIs with associated mutations: T66I, G118R and E138K; 7 cases(1.7%, 7/408) were resistant to nucleoside reverse transcriptase inhibitors(NRTIs) with major mutations: M184V/I, D67N for NRTIs; 5 cases(1.2%, 5/408) were resistant to non-nucleoside reverse transcriptase inhibitors(NNRTIs) with major mutation: K103N; 1 case(0.2%, 1/408) was resistant to protein inhibitors (PIs), and associated mutations was I50L.1 case was resistant simultaneously to INIs, NRTIs and NNRTIs, Dual-class mutations were observed in 4 cases(3 cases to NRTIs and NNRTIs, 1 case to PIs and NNRTIs).Conclusions:The study result showed that the drug resistance rate of INIs was low in Hunan province, however, the INIs drug resistance surveillance was necessary.

Objective:To understand the status of HIV-2 infection and the epidemiological characteristics of HIV-2 infected individuals in Hunan province.Methods:The stock blood samples with HIV-2 indicative bands in the original records of 15 AIDS confirmation laboratories in Hunan province from 2003 to 2020 were collected. The samples were tested by Mikrogen GmbH (HIV 1+ 2) antibody comfirmatory kit (Germany). And meanwhile the epidemiological data of the first HIV test and the epidemiological follow-up records of the cases who could be contacted were collected.Results:A total of 378 samples with HIV-2 indicative bands were collected, involving 363 individuals. Serological test result showed that 326 cases (89.81%) were HIV-1 antibody positive, 18 cases (4.96%) were HIV-2 antibody positive, 12 cases (3.31%) were HIV antibody positive but could not be typed, 6 cases (1.65%) were HIV antibody uncertain and 1 case (0.28%) was negative for HIV-1/2 antibody. There were 15 cases from Xiangtan city and the other 3 cases from Changsha city among the 18 HIV-2 antibody positive individuals. The result of epidemiological investigation showed that there were 12 males (66.67%) and 6 females (33.33%), with a male-to-female ratio of 2∶1. The average age of the first detection was (56.94±12.52) years, in the males the age ranged from 33 to 76 years and in the females the age ranged from 44 to 66 years. The male age span was larger than that of the female. There were 10 married cases (55.56%) including one couple, 6 divorced or widowed cases (33.33%), 2 cases (11.11%) with unknown marital status. There were 13 cases from hospital monitoring (72.22%), 2 cases each from prison detention and blood screening in volunteer donors (22.22%), and 1 case from spouse screening (5.56%). These samples were tested for the first time in 2005, 2009, 2013, 2014, 2018, and 2020, with 1 case each, 6 cases in 2016, 2 cases in 2017, and 4 cases in 2019. By the end of 2020, 9 cases were alive and 9 cases were dead or missing. Three cases have accepted antiviral treatment. Of the 7 cases that could be followed up, all admitted heterosexual transmission, 6 cases denied any history of traveling abroad, and 1 case denied any history of foreign sexual intercourse.Conclusions:HIV-2 epidemic has lasted for a long time in Hunan province, local infection is not an occasional case, and there is a possibility of long-term clustering epidemic spread. This suggests that the country should strengthen the surveillance, detection, diagnosis and treatment of HIV-2 research and pay attention to the risk of HIV-2 transmission.

Drosophila neural development undergoes extensive chromatin remodeling and precise epigenetic regulation. However, the roles of chromatin remodeling in establishment and maintenance of cell identity during cell fate transition remain enigmatic. Here, we compared the changes in gene expression, as well as the dynamics of nucleosome positioning and key histone modifications between the four major neural cell types during Drosophila neural development. We find that the neural progenitors can be separated from the terminally differentiated cells based on their gene expression profiles, whereas nucleosome distribution in the flanking regions of transcription start sites fails to identify the relationships between the progenitors and the differentiated cells. H3K27me3 signal in promoters and enhancers can not only distinguish the progenitors from the differentiated cells but also identify the differentiation path of the neural stem cells (NSCs) to the intermediate progenitor cells to the glial cells. In contrast, H3K9ac signal fails to identify the differentiation path, although it activates distinct sets of genes with neuron-specific and glia-related functions during the differentiation of the NSCs into neurons and glia, respectively. Together, our study provides novel insights into the crucial roles of chromatin remodeling in determining cell type during Drosophila neural development.

Objective To investigate the clinical value of CT lymphangiography (CTL) combined with direct lymphangiography (DLG) in primary intestinal lymphangiectasia (PIL). Methods Sixteen patients diagnosed as PIL with intestinal enteroscopy were recruited in this retrospective study. All the patients were performed DLG and CTL one week before exploratory laparotomy. Subjective assessment in DLG included weak lymphatic fluid drainage, lymphangiectasia, lymphatic reflux, fistula and thoracic outlet reflux or obstruction. While for CTL combined with DLG, the intestinal and extra-intestinal lesions were evaluated, including lymph node, edema, lymphangiectasia and abnormal distribution, fistula, and lymphangiomatosis. All the diagnosis was compared with intestinal endoscopy results. Results For DLG, 16 weak lymphatic fluid drainages, 9 lymphangiectasia, 1 fistula with abdomen and 14 thoracic outlets weak lymphatic fluid drainage or obstruction were found. For DLG combined with CTL, 16 intestinal lumens dilatation and 14 circumferential intestinal thickening were found in intestinal lesions. While for extra-intestinal lesions, the imaging features included edematous findings (12 in mesentery, 7 ascites only, 2 hydrothorax and ascites, and 3 pericardial, thoracic and abdominal effusions), abdominal lymph nodes (6 cases), lymphangiectasia and abnormal distributions (14 cases), fistulas (lymph-intestinal luminal fistula in 4 cases, and lymph-abdominal fistula in 3 cases), lymphangiomatosis (3 cases), and thoracic duct outlet dysfunction and reflux (14 cases).The number of cases diagnosed as intestinal lymphangiectasia, intestinal luminal lymph exudation and lymph fistula were 16, 10 and 6 with intestinal endoscopy, while the number were 11, 0, and 4 with CTL combination with DLG. Conclusion Combination of CTL with DLG is valuable in the diagnosis of PIL.

Objective To evaluate the value of imaging examinations in the treatment of lymphangioleiomyomatosis (LAM) with chylothorax by thoracic duct extremity exploration.Methods Data of 34 LAM with chylothorax confirmed by pathology and clinical diagnosis were retrospectively analyzed.All patients underwent 99Tcm-DX lymphoscintigraphy and CT lymphangiography (CTL).Thoracic duct lesion types of 99Tcm-DX lymphoscintigraphy were type Ⅰ (abnormal concentration pattern),type Ⅱ (ectopic drainage pattern),and type Ⅲ (without image or transient image pattern).The type Ⅰ and type Ⅱ were diagnosed as thoracic duct abnormalities.Thoracic duct lesion types of CTL were type Ⅰ (dilatation pattern),type Ⅱ (distal obstruction pattern),type l (truck constriction pattern),type Ⅳ (ectopic drainage pattern),and type Ⅴ (no-display pattern).Type Ⅰ-Ⅳ were diagnosed as thoracic duct abnormalities.Consistency of displaying thoracic duct abnormalities between 99Tcm-DX lymphoscintigraphy and CTL was evaluated.Results The thoracic duct abnormalities in 99Tcm-DX lymphoscintigraphy were 58.82％ (20/34;type Ⅰ in 17,type Ⅱ in 3,type Ⅲ in 14),and in CTL were 73.53％ (25/34;type Ⅰ in 15,type Ⅱ in 3,type Ⅲ in 5,type Ⅳ in 2,type Ⅴ in 9).The consistency of CTL and 99Tcm-DX lymphoscintigraphy for detecting thoracic duct abnormalities was good (Kappa=0.679).In CTL thoracic duct types,type Ⅰ and Ⅱ were operated by thoracic duct-venous anastomosis or thoracic duct extremity release operation,type Ⅲ was operated by thoracic duct adhesion or compression band release operation,operative approach and method were chosen according to the abnormal thoracic duct flow path in type Ⅳ,type Ⅴ was took conservative treatment.Conclusion CTL is superior to 99Tcm-DX lymphoscintigraphy,which can clearly display the type of thoracic duct lesion and provide imaging informations to choose operation methods in thoracic duct exploration treatment for LAM with chylothorax.

OBJECTIVETo determine the prevalence rates of nucleotide reverse-transcriptase inhibitor (NRTI) and nonnucleoside reverse transcriptase inhibitor (NNRTI)TDRs among HIV-1 ART-naїve patients in Hunan province using the ultra deep sequencing (UDS) technique.

METHODSART-naїve subjects diagnosed in Hunan between 2010 and 2011 were evaluated by both UDS technique and Sanger sequencing techniques, to the 1% variant level. Mutations were scored using the Stanford HIVdb algorithm to infer the status on drug resistance.

RESULTSUDS method was performed on 90 ART-naїve subjects that seeking service of care, in Hunan. In total, 42.2% (38/90) of the subjects showed major NRTI or nonnucleoside reverse transcriptase inhibitor NNRTI TDRs by UDS technique, at a HIV variant frequency level of ≥1%, 15.6% (14/90) showed NRTI TDR, 16.7% (15/90) showed a major NNRTI TDR and 10% (9/90) were both resistant to NRTI and NNRTI when variants were analyzed by Stanford HIVdb.

CONCLUSIONART-naїve subjects from Hunan province, which had been predominately infected by subtype AE, would frequently possess HIV variants with NRTI/NNRTI TDRs that would affect the use of first line ART in the region, identified by the UDS technique. Further studies were needed to describe the prevalence of TDRs and to gather related information.

Anti-HIV Agents ; therapeutic use ; China ; Drug Resistance, Viral ; genetics ; HIV Infections ; drug therapy ; virology ; HIV-1 ; genetics ; High-Throughput Nucleotide Sequencing ; Humans ; Mutation ; Prevalence ; Reverse Transcriptase Inhibitors ; analysis

Objective To determine the prevalence rates of nucleotide reverse-transcriptase inhibitor (NRTI) and nonnucleoside reverse transcriptase inhibitor (NNRTI) TDRs among HIV-1 ART-na?ve patients in Hunan province using the ultra deep sequencing(UDS) technique. Methods ART-na?ve subjects diagnosed in Hunan between 2010 and 2011 were evaluated by both UDS technique and Sanger sequencing techniques,to the 1%variant level. Mutations were scored using the Stanford HIVdb algorithm to infer the status on drug resistance. Results UDS method was performed on 90 ART-na?ve subjects that seeking service of care,in Hunan. In total,42.2%(38/90)of the subjects showed major NRTI or nonnucleoside reverse transcriptase inhibitor NNRTI TDRs by UDS technique,at a HIV variant frequency level of≥1%,15.6%(14/90)showed NRTI TDR,16.7%(15/90) showed a major NNRTI TDR and 10%(9/90)were both resistant to NRTI and NNRTI when variants were analyzed by Stanford HIVdb. Conclusion ART-na?ve subjects from Hunan province, which had been predominately infected by subtype AE,would frequently possess HIV variants with NRTI/NNRTI TDRs that would affect the use of first line ART in the region,identified by the UDS technique. Further studies were needed to describe the prevalence of TDRs and to gather related information.

Objective To investigate the diagnostic values of direct lymphangiography for the thoracic duct outlet obstruction.Methods The image data of direct lymphangiography were retrospectively analyzed in 124 patients with lymphedema,Chylothorax,chylous ascites,chyluria and intestinal lymphangiectasis,and compared with the results of neck thoracic duct surgical exploration,2 radiologists reviewed DLG DSA images in a double blind manner.The number of neck stem,subclavian stem,bronchialmediastinal stem and TD terminal into blood obstruction on the operation side showed by DLG were assessed using Kappa analysis.Results Of 124 patients,80 patients had the left cervical lymphatic stem reflux on DLG,75 patients with the left subclavian lymphatic stem reflux,30 patients with the left bronchial-mediastinal lymphatie stem reflux,118 patients showed the thoracic duct outlet barrier into the blood.The consistency rate of DLG were 89.9％ (80/89),92.6％ (75/81),90.9％ (30/33) and 95.2％ (118/124) compared with the neck thoracic duct surgical exploration.Tow radiologists had a high degree of diagnostic consistency (K =0.82,P ＜ 0.05).In addition,114 patients (91.9％) had tortuous,dilated waist lymphatic stem,only 10 patients (8.1％) were normal.The cisterna chyli reflux were found in 92 patients (74.2％),intestinal stem reflux in 16 patients (12.9％),reflux to the kidney area in 11 patients (8.9％),to the pericardium reflux in 5 patients (4.0％),vaginal lymphatic leakage in 7 patients (5.6％),retroperitoneal lymph leakage in 2 patients (1.6％),pleural lymphatic leakage in 3 patients (2.4％),tracheal lymph leakage in 1 patient (0.8％).Conclusion Direct lymphangiography has a high consistency with the cervical thoracic duct surgical exploration in displaying thoracic duct outlet obstruction.

Objective Toinvestigatethevalueofimagingdiagnosisofthe lymphangioleiomyomatosis( LAM ).MethodsFifteen patients with LAM confirmed by pathological assessment were analyzed retrospectively for radiologic findings.They had chest radiograph, chest highresolution CT (HRCT),abdominal CT, direct lymphangiography(DLG), chest CT and abdominal CT after DLG.Results Chest radiograph findings included normal (1),increasing of lung markings (3),disseminated honeycomb or reticular pattern ( 11 ), pneumothorax ( 2 ), and pleural effusion ( 14 ). Chest conventional CT and HRCT showed typical imaging manifestation of PLAM in all cases, including sporadic or disseminated cysts in bilateral lungs. According to the grading standard of pulmonary disease made by Avila et at, there were 3 cases in grade Ⅰ , 5 cases in grade Ⅱ and 7 cases in grade Ⅲ . Fourteen of 15 patients with LAM had positive abdominal CT findings in retroperitoneum and pelvic cavity. Common abdominal CT findings included cystic lymphangioma in 9 of 14 patients, lymphangiomyoma in 13 and both coexisting in 7.One of the14patients alsohadhepaticlipomaandangiomyolipomas.Onepatienthadrenal angiomyolipomas; and one patient had hysteromyoma. All 15 cases underwent DLG, 1 cases had lymphatic obstruction in the lumbar 3 level, the remaining 14 cases had varying degrees of thoracic duct stenosis, or obstruction. Neck trunk, subclavian trunk and bronchial trunk showed lymphatic reflux. On post-DLG CT,thoracic duet outlet obstruction was not demonstrated in 3 cases, the remaining 12 cases showed thoracic outlet obstruction, consistent with the DLG findings.Conclusion HRCT is a useful diagnostic method showing characteristic findings of PLAM. MSCT can help to detect abdominal LAM. DLG and MSCT after DLG have value in displaying obstruction site of thoracic duct or lymphatic trunks and provide guidance for operative treatment.

Proteolytic processing of viral polyproteins is indispensible for the lifecycle of coronaviruses. The main protease (M(pro)) of SARS-CoV is an attractive target for anti-SARS drug development as it is essential for the polyprotein processing. M(pro) is initially produced as part of viral polyproteins and it is matured by autocleavage. Here, we report that, with the addition of an N-terminal extension peptide, M(pro) can form a domain-swapped dimer. After complete removal of the extension peptide from the dimer, the mature M(pro) self-assembles into a novel super-active octamer (AO-M(pro)). The crystal structure of AO-M(pro) adopts a novel fold with four domain-swapped dimers packing into four active units with nearly identical conformation to that of the previously reported M(pro) active dimer, and 3D domain swapping serves as a mechanism to lock the active conformation due to entanglement of polypeptide chains. Compared with the previously well characterized form of M(pro), in equilibrium between inactive monomer and active dimer, the stable AO-M(pro) exhibits much higher proteolytic activity at low concentration. As all eight active sites are bound with inhibitors, the polyvalent nature of the interaction between AO-M(pro) and its polyprotein substrates with multiple cleavage sites, would make AO-M(pro) functionally much more superior than the M(pro) active dimer for polyprotein processing. Thus, during the initial period of SARS-CoV infection, this novel active form AOM(pro) should play a major role in cleaving polyproteins as the protein level is extremely low. The discovery of AOM(pro) provides new insights about the functional mechanism of M(pro) and its maturation process.
Coronavirus ; metabolism ; Cysteine Endopeptidases ; Endopeptidases ; metabolism ; Humans ; Peptides ; chemistry ; metabolism ; Polyproteins ; chemistry ; metabolism ; Protein Binding ; SARS Virus ; chemistry ; metabolism ; Viral Proteins