We wished to assess the role of chlamydia micro virus capsid protein Vp3 in recombinant molecules, chart its molecular evolution, screen the wild-type strain, and reveal its value in clinical research. Using a protein BLAST multiple-alignment program, we compared various strains of Chlamydia micro virus capsid protein Vp3 sequences. Using a "distance tree" of those results, we created a phylogenetic tree. We applied the Karplus-Schulz method of flexible-region analyses for highly conserved alignments of amino-acid sequences. Gamier-Robson and Chou-Fasman methods were employed to analyze two-level structures of sequences. The Emini method was used for analyses of the accessibility of surface epitopes. Studies of hydrophilic proteins were undertaken using Kyte-Doolittle and Hopp-Woods methods. Analyses of antigen epitopes helped to reveal the antigen index using the Jameson-Wolf method. All sequences in the six strains of chlamydia micro virus capsid protein Vp3 were highly conserved, with the main differences being between Vp3 protein in Chp1 and the other five strains of the micro virus. The viral strain of Vp3 protein was based mainly on micro-alpha helix structures, and multiple epitopes were noted in highly conserved regions. Vp3 protein was highly conserved structurally, and was an important protein of the chlamydiaphage capsid. Vp3 protein has a complicated molecular structure, highly conserved regions with strong immunogenicity, and has considerable research value.
Amino Acid Sequence ; Capsid Proteins ; chemistry ; genetics ; immunology ; Chlamydia ; genetics ; immunology ; Conserved Sequence ; Epitope Mapping ; Evolution, Molecular ; Molecular Sequence Data ; Recombination, Genetic

Objective To test cross immune responses induced in rhesus monkeys immunized with the recombinant major outer membrane protein(rMOMP).Methods Six rhesus monkeys were divided into three groups:the group vaccinated with purified rMOMP and Freund's adjutants,the group vaccinated with Freund's adjutants only and the control group vaccinated with PBS.All of the rhesus monkeys vaccinated intramuscularly at 0,2,4 weeks.Two weeks after the last time,The IFN-γand Chlamydia-specific antibody titers in sera,which were determined by ELISA,lymphocyte proliferation assay were performed by MTT,and observ the delayed hypersensitivity and in vitro neutralization assays.Results The result of the monkeys immunized with rMOMP and Freund's adjuvant:the specific immune responses can be observed.The in vitro neutralization and lymphocyte proliferation assays were observed better in the same group.Conclusion After being vaccinated with rMOMP,the monkeys can develop strong and effective Chlamydia-specific cross immune responses.

School bullying and campus violence is a widespread social problem in the world.School bullying is characterized by its repeatability and suddenness,which could make the victims suffering from both psychological and health damage,and even affect their personality growth.Government should pay close attention to the reduction and control of school bullying and campus violence by establishing school bullying emergency response system and preparedness plan.The school and teacher's role and legal responsibility in the service and management in schools should be cleared and defined.It is necessary to help teachers conduct early detection and intervention for school bullying,conduct morality,mental health and legal educations in students to teach them to act according to the law and protect themselves according to the law and help them identify and avoid risks,encourage the establishment of rescue facility and web of anti-school bullying by non-government organizations,and set hotline for school bullying incident to reduce the incidence of school bullying.

Objective To investigate the incidence of live births with Down syndrome(DS) in Sanya city as well as to discuss the related risk factors .Methods The object of investigation was the 25 032 infants who were born in Sanya city ,then the children with clinical suspected of DS were confirmed by peripheral blood chromosome analysis .Meanwhile ,the clinical data of children and their parents were collected ,calculating the incidence of DS in this region and analyzing the related risk factors .Results In this region , the total incidence of DS was 0 .959‰ ,the incidences in the Han nationality and Li nationality were 0 .834‰ ,1 .442‰ respectively . The mean age of DS children′s mother was 29 .8 years old ,and the incidence of DS increased as their mother grew older .Before pregnancy ,DS children′s mother who had an exposure history of toxicant and drugs accounted for 45 .8% .Conclusion The inci‐dence of DS in Sanya is quite high ,so it is extremely essential to strengthen the women′s pregnancy Down syndrome screening and improve the prenatal diagnosis in this region .

Objective To investigate the curative effect of cisplatin combined with gemcitabine and cisplatin plus pacli-taxel in the treatment of advanced non-small cell lung cancer. Methods All 100 patients with non-small cell lung can-cer in our hospital from January 2012 to January 2014 in our hospital, randomly divided into observation group (gemc-itabine+cisplatin, GP group) and control group (paclitaxel+cisplatin, TP group), each group had 50 cases, two groups of patients every 21 days for a cycle, observed curative effect after 2 cycles, toxic and adverse reaction were compared between the two groups. Results The effective rate of the observation group after treatment was 42.0%(21/50),was higher than that of the control group 40% (20/50), but there was no significant difference (P>0.05). Conclusion Cura-tive effect of cisplatin combined with gemcitabine and cisplatin plus paclitaxel in the treatment of advanced non-small cell lung cancer is similar, the poisonous side effect is different and can be tolerated, for the treatment of advanced non-small cell lung cancer treatment plan better.

Objective To investigate the impact of clinical factors on survival in patients receiving concurrent chemotherapy and three?dimensional radiotherapy ( 3DRT) for stage IV non?small cell lung cancer ( NSCLC) . Methods A total of 203 patients were enrolled in a prospective clincial study from 2008 to 2012, and among these patients, 178 patients were eligible for analysis of clinical factors. All patients were treated with platinum?based doublets chemotherapy, with a median number of chemotherapy cycles of 4( 2?6 cycles) and a median dose of 3DRT of 60?3 Gy (36?0?76?5 Gy).The Kaplan?Meier method was used to calculate overall survival ( OS) rates, the log?rank test was used to compare survival rates between groups, and the Cox regression model were used for multivariate analysis. Results The 1?, 2?, and 3?year overall survival rates were 56%, 16%, and 10%, respectively, and the median survival time was 13 months (95% CI=11?500?14?500). The univariate analysis showed that platelet count ≤221×109/L, neutrophil count ≤5.2×109/L, white blood cell count<7×109/L, and improvement in Karnofsky Performance Scale ( KPS) after treatment significantly prolonged OS ( P=0?000,0?022,0?003, and 0?029) , and metastasis to a single organ and hemoglobin≥120 g/L tended to prolong OS (P=0?058 and 0?075). The multivariate analysis showed that white blood cell count<7×109/L, platelet count ≤221×109/L, and improvement in KPS after treatment were beneficial to OS ( all P<0?05) . Conclusions White blood cell count and platelet count before treatment and KPS after treatment are prognostic factors for patients with stage IV NSCLC receiving concurrent chemotherapy and 3DRT. Clinical Trial Registry ClinicalTrials. gov, registration number:ChiCTRTNC10001026.

Objective:To understand the prevalence of arboviruses in mosquito samples in Xichang City, Sichuan Province, and enrich the data of arbovirus activity and genetic characteristics in southwestern Sichuan Province.Methods:In June 2018, the nucleic acid was extracted from Culex tritaeniorhynchus mosquitoes collected from different pigsties in three villages and suburbs of Xichang City. The specific primers of Yunnan orbivirus, Banna virus, Tibet orbivirus (S7, S10), Flavivirus and alphavirus were used for quantitative polymerase chain reaction examination, and the positive product was cloned for sequencing analysis. Results:A total of 9 012 mosquitoes were collected, of which Cx. tritaeniorhynchus was the dominant species. A number of 88 batches of these mosquitoes were amplified, and 2 strains of Japanese encephalitis virus (JEV), 7 strains of Banna virus (BAV), 7 strains of Tibet orbivirus (TIBOV) and 1 strain of Yunnan orbivirus virus (YOUV) were detected, respectively. By the results of cluster analysis and evolutionary tree analysis, the 17 newly found virus strains were close to the Yunnan isolates, and 2 JEV strains were located in the GI-b clade. The other 7 strains of BAV were A2 evolutionary clades. Of the 7 TIBOV plants, 6 were located in the same clade. One TOUV was in the same clade as the Yunnan strain. Conclusions:Culex tritaeniorhynchus mosquitoes in Xichang city might carry JEV, BAV, YOUV and TIBOV, among them JEV was GI-b type and BAV was A2 type. The results provide data supporting the detection and analysis of arboviruses in Xichang city.

OBJECTIVETo study the cytomorphologic features of anaplastic lymphoma kinase (ALK)-rearranged pulmonary adenocarcinoma.

METHODSThe morphologic features in 153 pulmonary adenocarcinoma cytology specimens encountered during the period from September, 2011 to April, 2015 in Shanghai Cancer Hospital were retrospectively reviewed. Fluorescence in-situ hybridization (FISH) and/or immunohistochemistry (Ventana D5F3) for ALK gene rearrangement were carried out. The samples studied included 34 pleural effusion specimens, 40 endobronchial ultrasound-guided transbronchial needle aspirates (EBUS-TBNA) and 79 fine needle aspirates of palpable masses on body surface.

RESULTSThirty-nine cases (25.5%) of ALK-rearranged samples were identified by FISH and/or immunohistochemistry, including 3 cases diagnosed by FISH and 36 cases by both technologies. The median age of the ALK-positive group was 50 years, significantly younger than that of the ALK-negative group (60 years old, P = 0.002). Only 4 of the ALK-positive patients were smokers, which was significantly less than that of the ALK-negative group (P < 0.01). In ALK-positive group, 3 cases showed cribriform pattern with prominent nucleoli, 3 cases showed cribriform pattern with mucin-rich cells and 8 cases showed extracellular mucus with mucin-rich cells. The above cytomorphologic patterns were significantly less common in ALK-negative tumors (P < 0.01).

CONCLUSIONSALK-rearranged lung adenocarcinoma is associated with certain distinctive morphologic patterns, including cribriform architecture, presence of prominent nucleoli, mucin-rich cells and extracellular mucus, which can be observed in cytology specimens (including conventional smears and cell block sections). These findings, when combined with clinical features, may give clues to detection of ALK-positive cases.

Adenocarcinoma ; genetics ; pathology ; Biopsy, Fine-Needle ; China ; Gene Rearrangement ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; genetics ; pathology ; Receptor Protein-Tyrosine Kinases ; genetics ; Retrospective Studies

Objective To isolate the Japanese encephalitis virus carried by Culex tritaeniorhynchus in Dongchuan District of Yunnan Province and analyze its molecular characteristics, so as to provide insights into the prevention and control of Japanese encephalitis in Yunnan Province. Methods Mosquito specimens were collected using mosquito-trapping lamps from pig farms in Batang Village and Xiaoxin Village, Dongchuan District, Kunming City, Yunnan Province in July 2016, and the mosquito species was identified according to the mosquito morphology. Then, 60 to 100 mosquitoes of each species served as a group and were ground. Baby hamster kidney-21 (BHK-21) cells and Aedes albopictus clone C6/36 cells were used for virus isolation, and positive isolates were identified using flavivirus primers. The positive isolates were amplified using reverse transcription polymerase chain reaction (RT-PCR) assay with 15 pairs of specific primers covering the full length of the genotype I Japanese encephalitis virus, and DNA sequence assembly was performed using the software SeqMan in the DNASTAR package. The obtained sequences were aligned with the complete sequences of 38 Japanese encephalitis virus downloaded from the GenBank with the software MegAlign, and the nucleotide and amino acid homology analyses of the obtained sequences were performed. The difference in amino acid sites was analyzed with the software GeneDoc, and phylogenetic trees were created based on the sequences of the coding region and E protein of the isolated Japanese encephalitis virus with the software Mega X. In addition, the secondary and tertiary structures of the E protein of the Japanese encephalitis virus were predicted using the online tool SOPMA and the software Swiss-Model. Results A total of 5 820 mosquitoes were collected and 3 843 Cx. tritaeniorhynchus (66.03%) were identified according to the mosquito morphology. A positive virus isolate, termed YNDC55-33, was isolated from Cx. tritaeniorhynchoides following batches of virus isolation from mosquito specimens, and cytopathic effect was observed following inoculation into BHK-21 and C6/36 cells. The YNDC55-33 virus isolate was successfully amplified with the flavivirus primes, and a long sequence containing 300 nucleotides was obtained. Following sequence alignment using the BLAST tool, the sequence of the YNDC55-33 virus isolate had high homology with that of the genotype I Japanese encephalitis virus. A long sequence with 10 845 nucleotides in length, which encoded 3 432 amino acids, was obtained by splicing the full sequence of the YNDC55-33 virus isolate. Phylogenetic analysis based on the whole-genome sequence and E gene sequence of the YNDC55-33 virus isolate showed that the new YNDC55-33 virus isolate was most closely related to the genotype I Guizhou isolate (GenBank accession number: HM366552), with nucleotide homology of 98.5% and amino acid homology of 99.4%, and the YNDC55-33 virus isolate shared 97.96% ± 0.33% nucleotide homology and 99.35% ± 0.08% amino acid homology with other genotype I Japanese encephalitis virus isolates, and < 90% nucleotide homology and < 98% amino acid homology with other genotypes of Japanese encephalitis virus. The YNDC55-33 virus isolate and the live attenuated virus vaccine candidate SA14-14-2 isolate differed at 16 amino acid sites on E gene, and 7 out of 8 key amino acid sites related to neurovirulence. The secondary and tertiary structures of the E protein of the YNDC55-33 virus isolate were predicted to be characterized by random coils. Conclusions A genotype I Japanese encephalitis virus was isolated from Cx. tritaeniorhynchus in Dongchuan District, Kunming City. This virus isolate and the live attenuated virus vaccine candidate SA14-14-2 isolate does not differ at antigenic epitopes-related key amino acid sites, and the major protein structure of the virus isolate is random coils. This study adds new data for the epidemiological distribution of Japanese encephalitis virus in Yunnan Province, which may provide insights into the prevention and control of Japanese encephalitis in the province.