Objective:To analyze the clinical characteristics of dengue fever patients, summarize the course and characteristics of the disease, and analyze the risk factors that affect the condition.Methods:Retrospective collection of general information, clinical symptoms, medical history, laboratory tests, prognosis and other clinical data of dengue fever patients that admitted to Jinghong First People's Hospital and severe dengue fever patients at People's Hospital of Xishuangbanna Dai Autonomous Prefecture from June to December 2023 was conducted using a case report form (CRF). According to the diagnostic criteria of the World Health Organization (WHO), patients were divided into dengue fever group, dengue fever with warning signs group, and severe dengue fever group. The differences in clinical data between different groups of patients were analyzed and compared. Binary multiple factor Logistic regression analysis was used to explore the risk factors affecting the severity of dengue fever in patients. Receiver operator characteristic curve (ROC curve) was drawn to analyze the predictive value of prediction models constructed for various risk factors for severe dengue fever. Subgroup analysis was performed on the prognosis of severe dengue fever patients, and the differences in clinical data between two groups of patients with different prognoses were compared. Binary multivariate Logistic regression analysis was used to explore the risk factors affecting the prognosis of severe dengue fever patients. ROC curve was drawn to analyze the predictive value of prediction models constructed for various risk factors on the prognosis of severe dengue fever patients.Results:A total of 2 264 patients were included, including 499 cases in the dengue fever group, 1 379 cases in the dengue fever with warning signs group, and 386 in the severe dengue fever group (43 deaths and 343 survivors). The most common symptom of dengue fever patients was fever (94.70%), followed by muscle soreness (70.54%), headache (63.12%), fatigue (58.92%), and chills (46.02%). Compared with the dengue fever group and the dengue fever with warning signs group, the ratio of thalassemia and the levels of cardiac troponin (cTnI, cTnT), MB isoenzyme of creatine kinase (CK-MB), and myoglobin were significantly increased in patients with severe dengue fever group, albumin (Alb) was significantly decreased in patients with severe dengue fever group. The levels of cTnT and myoglobin in patients with dengue fever with warning signs group were significantly higher than those in the dengue fever group, and the level of Alb in patients with dengue fever with warning signs group was significantly lower than that in the dengue fever group, the differences were statistically significant (all P < 0.05). Binary multivariate Logistic regression analysis showed that thalassemia [odds ratio ( OR) = 6.214, 95% confidence interval (95% CI) was 2.337-16.524, P < 0.001], Alb ≤ 36 g/L ( OR = 6.297, 95% CI was 4.270-9.286, P < 0.001), and cTnT levels ( OR = 1.008, 95% CI was 1.002-1.015, P = 0.016) were risk factors for severe dengue fever. ROC curve analysis showed that the area under the ROC curve (AUC) for predicting severe dengue fever based on the prediction models constructed for the above risk factors was 0.856, with the best predictive value of 0.067, sensitivity of 67.1%, and specificity of 99.4%. In the subgroup analysis of patients with severe dengue fever, compared with the survival group, the levels of hematocrit (HCT), cTnT, and CK-MB in the death group patients were significantly increased, while the level of Alb was significantly decreased, and the differences were statistically significant. Binary multivariate Logistic regression analysis showed that Alb ( OR = 0.839, 95% CI was 0.755-0.932, P = 0.001), HCT ( OR = 1.086, 95% CI was 1.010-1.168, P = 0.025), elevated troponin level ( OR = 10.119, 95% CI was 2.596-39.440, P < 0.001), and CK-MB ( OR = 1.081, 95% CI was 1.032-1.133, P < 0.001) were risk factors for mortality in patients with severe dengue fever. ROC curve analysis showed that the AUC for predicting death in severe dengue fever patients based on the prediction models constructed for the above risk factors was 0.881, with the best predictive value of 0.113, sensitivity of 75.0%, and specificity of 88.9%. Conclusion:Thalassemia, Alb≤36 g/L, and cTnT level are risk factors for severe dengue fever, while HCT level, Alb level, CK-MB level, and elevated troponin level are risk factors for death in patients with severe dengue fever.

Objective:To analyze the biological characteristics, phylogeny and the taxonomic status of strain 7712 (=CGMCC 1.17031=NBRC 113783=KCTC 15766) isolated from a clinical blood sample.Methods:Strain 7712 was identified by the cultural properties, cellular and colonial morphology, physiological and biochemical reactions, matrix-assisted laser desorption ionization time-of-flight mass spectrometry system, and genome correlation index analysis. The genomic phylogenetic tree was construct to analyze the taxonomic position. The virulence factors and resistance genes of strain 7712 and related strains were then compared by the online virulence factor database and online comprehensive antibiotic research database respectively.Results:Strain 7712 was urease negative, gram-negative nonfermenters, which was identified as Ochrobactrum anthropi by VITEK GN card. The 16S rRNA gene analysis showed that the strain was closely related to the members of genera Ochrobactrum and Brucella. The phylogenetic tree showed that strain 7712 was clustered together with Ochrobactrum teleogrylli LCB8 T and Ochrobactrum haematophilum CCUG 38531 T, along with genus Brucella and other Ochrobactrum species. The genome relatedness indexes analysis showed that the average nucleotide identity between strain 7712 and Ochrobactrum teleogrylli LCB8 T was 98.16%, which was higher than the threshold for prokaryotic species. Genetic prediction showed that strain 7712 carried several virulence-related genes and resistance-related genes, of which the existence of OCH gene might be responsible to the resistance to cephalosporin. Conclusions:A case of human infection caused by Ochrobactrum teleogrylli is identified, which would help promote the understanding of biodiversity of genus Ochrobactrum.

Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which causes great economic losses. At the moment, no effective neutralizing antibody is available for scientific research and treatment. Therefore, developing a method for screening the neutralizing monoclonal antibodies is of great significance for the prevention and treatment of PRRSV and the screening of antigen sites. Monoclonal antibodies have been widely used in the treatment and diagnosis of many human and animal diseases. Therefore, screening effective neutralizing antibodies for different pathogens is an urgent task. Among the methods for monoclonal antibody screening, B cell immortalization is an effective method to obtain neutralizing monoclonal antibody. Specifically, in this study, the bcl-6 and bcl-xl genes were connected by f2a and then the yielded product was ligated to a vector for retrovirus packaging. The swine lymphocytes immunized with PRRSV were infected the yielded mature viruses and cultured in the complete medium containing CD40L and IL21 cytokines. Then, CD21 was used as the marker to screen B cells with the magnetic bead method. Finally, monoclonal B cells were obtained and the secretion of antibodies was tested. The results showed that the plasmid, either being transfected alone or with the packaged plasmids, could be expressed, and that the packaged retrovirus could infect the cells. Moreover, the infected lymphocytes secreted antibodies, so did the screened B cells. Therefore, the method for screening monoclonal antibody against PRRSV was successfully established.
Animals ; Antibodies, Monoclonal ; Antibodies, Neutralizing ; Antibodies, Viral ; Humans ; Porcine Reproductive and Respiratory Syndrome/prevention & control* ; Porcine respiratory and reproductive syndrome virus/genetics* ; Swine

Objective To understand the epidemiological characteristics of foodborne disease outbreaks in Zhaotong City of Yunnan Province from 2010 to 2019, and to provide a scientific basis for the prevention and control of foodborne disease outbreaks in Zhaotong City. Methods Data were collected from National Foodborne Diseases Surveillance Network during 2010-2019, and a descriptive analysis of the data was carried out. Results A total of 82 foodborne disease outbreaks were reported from 2010 to 2019, involving 2 060 cases and 40 deaths. In 2016, there were 12 (accounting for 14.63%) foodborne disease outbreaks. Foodborne disease outbreaks displayed seasonal pattern, and mainly occurred from July to October. Families were the main places of foodborne disease outbreaks, accounting for 63.41%. Poisonous mushroom was the main food for outbreaks, which caused 39 outbreaks of foodborne diseases (accounting for 47.56%）. Twenty-six people died from eating wild mushroom, accounting for 65.00% of the total deaths. Drinking and eating by mistake were the main cause of the disease, leading to 47 cases (57.31%). Conclusions Summer and Autumn are the high-risk seasons for foodborne disease outbreaks in Zhaotong City. Eating and drinking by mistake are the main cause of the disease, and eating poisonous mushroom is the predominant reason. It is necessary to strengthen food safety education, to improve food safety awareness, and to strengthen the supervision of food safety in collective canteens and issue early warning intervention in time in the season of high incidence of foodborne diseases.

Antigenic purity is important for quality control of the foot-and-mouth (FMD) whole virus inactivated vaccine. The recommended method for evaluation the antigenic purity of FMD vaccine is to check the serum conversion to non-structural protein (NSP) 3AB antibody after 2 to 3 times inoculation of animals with inactivated vaccine. In this study, we developed a quantitative ELISA to detect the amount of residual 3AB in vaccine antigen, to provide a reference to evaluate the antigenic purity of FMD vaccine. Monoclonal antibody (Mab) of NSP 3A and HRP-conjugated Mab of NSP 3B were used to establish a sandwich ELISA to quantify the NSP 3AB in vaccine antigen of FMD. Purified NSP 3AB expressed in Escherichia coli was serially diluted and detected to draw the standard curve. The detectable limit was determined to be the lowest concentration of standard where the ratio of its OD value to OD blank well was not less than 2.0. Results: The OD value was linearly corelated with the concentration of 3AB protein within the range between 4.7 and 600 ng/mL. The correlation coefficient R² is greater than 0.99, and the lowest detectable limit is 4.7 ng/mL. The amount of 3AB protein in non-purified inactivated virus antigen was detected between 9.3 and 200 ng/mL depending on the 12 different virus strains, whereas the amount of 3AB in purified virus antigen was below the lowest detectable limit. The amount of 3AB in 9 batches of commercial FMD vaccine antigens was between 9.0 and 74 ng/mL, whereas it was below the detectable limit in other 24 batches of commercial vaccine antigens. Conclusion: the sandwich ELISA established in this study is specific and sensitive to detect the content of 3AB protein in vaccine antigen of FMD, which will be a useful method for evaluation of the antigenic purity and quality control of FMD inactivated vaccine.
Animals ; Antibodies, Viral ; Enzyme-Linked Immunosorbent Assay ; Foot-and-Mouth Disease/prevention & control* ; Foot-and-Mouth Disease Virus ; Viral Nonstructural Proteins/genetics* ; Viral Vaccines

Objective To clarify the characteristics and postoperative benefit of gallbladder carcinoma in elderly patients (≥ 65 years old).Methods Two hundred and seventy-three patients of gallbladder carcinoma were collected,who were treated intent resection from January 2004 to December 2012 in the Department of Hepatobiliary Surgery,Eastern Hepatobiliary Hospital,Second Military Medical University.More than 65 years old was defined as the elderly,else was defined as the younger.The clinical-pathological features and prognosis of 85 elderly patients(elderly group) and 188 younger patients (younger group) were retrospectively analyzed.The survival of patients were followed up by telephone or outpatient.The incidence of hypertension,incidence of diabetes,TNM staging,and median CA19-9 were compared between the two groups.Continuous variables using a two-sample t test or Mann-Whitney U test.Categorical variables were compared by the Chi-square test or Fisher probability method.The survival curve was drawn by the Kaplan-Meier method.The univariate analysis and multivariate analysis of prognosis were respectively done using the Log-rank test and COX regression model.Results The incidence of hypertension,incidence of diabetes,TNM stage ratio (Ⅲ + Ⅳ/Ⅰ + Ⅱ),and CA19-9 median in the elderly group were 30.6％,11.8％,27.6 and 69.3 U/ml,respectively.The differences in the younger group were 13.8％,4.8％,7.9 and 28.2 U/ml,respectively,with statistically significant difference between the two groups (all P ＜ 0.05).The incidence of complications was 54.1％ in the elderly group and 48.9％ in the younger group,with no significant difference between the two groups (P =0.302).The median survival of the elderly group was 28.01 months,and the median survival of the younger group was 36.20 months,with no statistical difference between the two groups (P =0.131).Cox analysis showed that independent prognostic risk factors for the elderly patients with gallbladder cancer included liver invasion (HR =2.386,95％ CI:1.379-4.127,P =0.002) and lymph node metastasis (HR =1.866,95 ％ CI:1.100-3.167,P =0.021).Conclusions Radical resection is safe and feasible for elderly patients with gallbladder carcinoma.Age is not a contraindication for surgery.Radical resection can get the same benefits as young people.Liver invasion and lymph node metastasis are independent risk factors affecting the prognosis of the elderly patients with gallbladder carcinoma.

Chinese traditional culture is the soil for the survival and development of traditional Chinese medi-cine,among them,the concept of A thing is valued if it is rare portal factions and master has influenced the development,communication and popularization of traditional Chinese medicine.But the spirit and thinking of harmony unity of heaven and man comparative state benevolence and self-cultivation and self-re-straint has played an active role in the development of traditional Chinese medicine.Thus this paper put forward the suggestions and opinions of strengthening the cultivation of humanistic spirit of higher traditional Chinese medi-cine talents:establishing a comprehensive evaluation standards of higher traditional Chinese medicine talents,es-tablishing scientific development concept to treat the inheritance and innovation of traditional Chinese medicine cul-ture,perfecting the curriculum provision of relevant professions,creating the conducive environment for the trans-mission of traditional Chinese medicine culture,hoping to provide a reference for the explanation of the relationship between traditional culture and traditional Chinese medicine.

Objective To identify and analyze the homology of Ochrobactrum isolated from clinical blood samples of children.Methods The 26 strains of Ochrobactrum anthropi were identified by Vitek 2 Compact and test strips of API 20 NE bacterial identification system.The biochemical phenotypes were identified by manual tests.The 16S rRNA and recA gene were amplified by PCR and sequenced.The drug sensitivity tests of Ochrobactrum anthropi were performed by Vitek 2 Compact and matched GN13 card.The homology was analyzed by pulsed field gel electrophoresis.Results Based on the identification of the instruments and the manual tests for biochemical phenotype,all the 26 experimental strains were Ochrobactrum anthropi.The results of sequencing for 16S rRNA and recA gene amplification products showed 25 strains were Pseudochrobactrum saccharolyticum and the other 1 was O.grignonensein.Drug sensitivity analysis showed that the all the 26 strains were resistant to aztreonam,but the sensitive rates to quinolones,aminoglycosides,trimethoprim sulfamethoxazole,four generation of cephalosporins and the antibiotics compound of piperacillin/tazobactam were all more than 80％.Pulsed field gel electrophoresis analysis showed that the 25 strains were highly homologous with differences of only 1 to 3 bands in fingerprint profiles.Conclusion Based on the biochemical phenotype and the sequencing of 16S rRNA and recA gene,the Ochrobactrum-like bacteria could be identified to the level of species.The highly homologous strains of Pseudochrobactrum saccharolyticum may be sourced from a clustered infection.

Objective To evaluate the application of time to positivity (TTP)in differential diagnosis of intracranial infection caused by coagulase-negative Staphylococcus (CNS).Methods One hundred and twenty-four adult patients with positive CNS isolated from cerebrospinal fluid (CSF)including 70 cases with intracranial infection and 54 cases of CSF contamination,who were admitted in Xiangya Hospital of Central South University during January to December 201 5,were retrospectively analyzed.The difference of TTP between two groups was compared,receiver operating characteristic (ROC)curve was analyzed and the area under the ROC curve (AUC)was calculated.The application of TTP in differential diagnosis of CNS infection was evaluated.SPSS 1 8.0 software was used to analyze the data.Results TTP in intracranial infection group was shorter than that in CSF contamination group [(23.5 ±7.5 )h vs. (37.6 ±1 0.5)h,t =-8.71 7,P =0.000].The AUC of TTP was 0.854.Taking the cut-off value of 27.94 h,the sensitivity,specificity,positive and negative predictive values in differentiation of two groups were 72.7%,91 .4%,90.0% and 72.2%,respectively.There were statistically differences in TTP of Staphylococcus epidermidis,Staphylococcus haemolyticus and Staphylococcus capitis between two groups (Z =-4.496,-2.322 and -2.399,respectively,P <0.05 or <0.01 ).Conclusion TTP can be used to discriminate early intracranial infection and CSF contamination caused by CNS,and also can identify intracranial infection caused by different categories of CNS.

Objective To test a snapback primer for the identification of Legionella and Legionella pneumophila ( L.pneumophila) strains in a one-step real-time PCR assay.Methods A novel primer was designed with a pair of genus-specific primers of Legionella strains.The species-specific probe sequences of L.pneumophila strains were linked at the 5′end of the reverse primer.The sensitivity and specificity of the novel PCR assay were tested with 43 types of Legionella and non-Legionella strains.The established PCR as-say was used to identify 186 wild Legionella strains isolated from 11 provinces of China and 15 environmental water samples.Results The amplicon melting peak of Legionella strains was detected at 85-86℃.The snapback melting peak of L.pneumophila was detected at 71℃.No melting peak of non-Legionella strains was detected.The sensitivity of the standard strains and simulated water samples were 1 ng/μl DNA tem-plates and (1×103-1×104 )/ml, respectively.186 Legionella strains and 44 L.pneumophila strains isolated from environmental water samples were successfully identified with the snapback primer.Twealve Legionella strains and 4 L.pneumophila strains were identified from 15 environmental water samples with the snapback primer as compared with 8 Legionella strains identified with the culture method.Conclusion The snapback primer mediated one-step PCR assay could be used for the identification of Legionella and L.pneumophila strains with the advantages of high specificity and sensitivity.