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.

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.

Objective:To analyze the application effect of integrated medical and nursing management model based on intelligent medical system in preventing postoperative breast cancer lymphedema.Methods:A total of 180 patients undergoing axillary lymph node dissection for breast cancer were selected in the Affiliated Hospital of Nantong University from July 2018 to August 2019. According to the random number table method, they were divided into treatment group and control group for 90 cases in each group, and finally completed the study: 86 cases in treatment group and 82 cases in control group. The control group was given routine health management, and the treatment group was given an integrated management model based on intelligent medical systems. After 6 months of follow-up, the two groups of patients were compared for their cognition of lymphedema, prevention behavior, incidence of lymphedema, and patient satisfaction.Results:The incidence, clinical manifestations, risk factors, prevention methods, and overall awareness rates of lymphedema in the treatment group were 82.56%(71/ 86), 84.88%(73/86), 83.72%(72/86), 83.72%(72/86), 83.72%(72/86), and the control group were 67.07%(55/82), 70.73%(58/82), 68.29%(56/82), 69.51%(57/82), 70.73%(58/82), the differences were statistically significant ( χ2 values were 4.046-5.508, P<0.05). The total scores of skin care, lifestyle, avoidance of upper limb compression, and prevention of lymphedema in the treatment group were (9.54±1.04), (30.45±2.45), (9.35±1.08), (58.92±8.20) points, and the control group were (8.12±1.32), (8.12±1.32), (8.74±1.14), (53.45±7.64) points, the differences were statistically significant ( t values were 3.561-7.764, P<0.01). The incidence of lymphedema in the treatment group was 9.30%(8/86), and that in the control group was 23.17%(19/82), the difference was statistically significant ( χ2 value was 5.985, P<0.05). Satisfaction was 95.35%(82/86) in the treatment group and 82.93%(68/82) in the control group, the difference was statistically significant ( χ2 value was 6.771, P<0.01). Conclusions:The integrated management of medical care and patients based on intelligent medical system can help improve the level of lymphedema cognition in patients with breast cancer surgery, promote the development of lymphedema prevention behavior, reduce the incidence of postoperative lymphedema, and improve patient satisfaction.

Objective:To analyze the clinical characteristics, therapeutic options and risk factors of mortality in patients with carbapenem-resistant Acinetobacter baumannii (CRAB) bloodstream infection, and to provide evidence for clinical treatment option and prognosis evaluation of CRAB bloodstream infections. Methods:A retrospective study was carried out in 224 patients with confirmed diagnosis of CRAB bloodstream infection during the period from January 2012 to December 2017 in West China Hospital, Sichuan University. The patients were divided into the death group and the survival group according to the survival status 28 days after collecting blood samples. The clinical features and therapeutic options of antibacterial drugs were reviewed. Student′s t test was used for analyzing normally distributed data and Mann-Whitney U test for non-normal data.Chi-square test was used for categorical variables. Univariate and multivariate logistic analysis were used to analyze the risk factors of mortality associated with CRAB bloodstream infection. Results:Among 224 cases of CRAB bloodstream infection, 121 cases died (54.02%). These patients were mainly in intensive care unit (ICU) and hematology department. The common underlying diseases were severe acute pancreatitis and severe cardiovascular events. The interleukin (IL)-6 level (median (interquartile range)) in the death group (480.40 ng/L (1 432.95 ng/L)) was higher than that of the survival group (107.05 ng/L (263.08 ng/L)), the difference was statistically significant ( Z=4.526, P<0.01). The procalcitionin (PCT) levels in the death group and the survival group were 3.81 μg/L (17.26 μg/L) and 2.12 μg/L (12.74 μg/L), respectively, with no difference between the two groups ( P>0.05). The death rate of empirical treatment with a single or more non-active antimicrobial agents was 57.14% (64/112), that of monotherapy with active agent was 45.68% (37/81), and that of combination therapy with at least one active drug was 64.52% (20/31). The differences had no statistical significance ( P=0.130). The logistic regression analysis showed that the risk factors of mortality associated with CRAB bloodstream infection were renal dysfunction (odds ratio ( OR)=2.181, P=0.024) and multiple organ dysfunction syndrome (MODS; OR=20.376, P<0.01). Conclusions:The fatality rate of patients with CRAB bloodstream infection is high. These patients with renal dysfunction or MODS have poor prognosis. In addition to early effective antibacterial therapy, individual comprehensive treatment should be implemented in order to improve the curative effect.

Objective? To investigate the training, operation and examination of nursing care for lymphedema in Jiangsu Province. Methods? We selected 99 nurses provided nursing care for lymphedema at 13 class Ⅲ hospitals of prefecture-level city in Jiangsu Province as subjects by purposive sampling. All of nurses were investigated with the self-designed questionnaire involving general information, application and training of nursing care for lymphedema, nursing operation and examination of lymphedema. Results? Among 99 nurses, 68 (68.69%) nurses could carry out training regularly, and 59 (59.60%) nurses participated in training on nursing care for lymphedema. The main causes failing to could carry out training regularly included the teaching staff and lack of related content. Conclusions? At present, the training of nursing care for lymphedema needs to be strengthened. There is a pressing need to formulate the unified standardized operation and examination standards so as to improve the quality of work and professional connotation of nursing care for lymphedema.

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.

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 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 observe the specific immune responses induced by the recombinant major outer membrane protein (rMOMP) vaccine against Chlamydia trachomatis E serotype in rhesus monkeys.[Methods] Six rhesus monkeys were equally divided into three groups:adjuvant and protein group vaccinated with purified rMOMP and Freund's adjuvants,adjuvant group immunized with Freund's adjuvants only,and control group immunized with phosphate buffer.All the rhesus monkeys were intramuscularly immunized in the triceps brachii for 3 times at a 2-week interval.Two weeks after the last vaccination,serum,vaginal wash and venous blood samples were collected from the rhesus monkeys,and lymphocytes were isolated from the blood samples.Enzyme linked immunosorbent assay (ELISA) was performed to determine the specific IgG antibody and interferon level in sera and secretory IgA (sIgA) level in wash samples,and methyl thiazolyl tetrazolium (MTT) assay to evaluate the proliferation of lymphocytes after stimulation with Chlaraydia trachomatis serotype E elementary bodies.Delayed hypersensitivity was observed in rhesus monkeys challenged by inactivated Chlamydia trachomatis serotype E elementary bodies.In vitro antibody neutralization assay was conducted with the serum from rhesus monkeys.Indirect immunofluorescenee was used to detect Chlamydia trachomatis in exfoliative vaginal cells from rhesus monkeys from week 1 to 10 after challenge with Chlamydia trachomatis.Data were statistically analyzed by using one-way analysis of variance and least significant difference (LSD) test with the SPSS 14.0 software.[Results] The adjuvant and protein group differed statistically from the adjuvant group and control group in the serum level of specific IgG antibody (1.718 ± 0.213 vs.0.841 ± 0.315 and 0.791 ±0.437,both P＜ 0.05),interferon ((1086 ± 121.730) ng/L vs.(409 + 53.440) ng/L and (162 ± 48.046) ng/L,both P＜ 0.05),lymphocyte proliferation index (7.012 ± 1.026 vs.4.473 ± 1.850 and 1A26 ± 1.104,both P＜0.01 ) and the diameter of nodus in delayed hypersensitivity assay ( ( 1 1 ± 2.134) mm vs.(3 ± 0.914) mm and 0,both P ＜ 0.01 ).After attack,the exfoliative cells kept positive for Chlamydia trachomatis in the adjuvant and protein group from week 1 to 5,and in the other 2 groups from week 1 to 10,but were negative in the adjuvant and protein group from week 6 to 10.[Conclusion] The rMOMP vaccine can induce a specific,protective,humoral and cellular immune response against Chlamydia tracbomatis in rhesus monkeys.

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.