Objective] To investigate the possible mechanism underlying the killing of Culex quinquefasciatus larvae by Lagenidium giganteum from the biochemical point of view. [Methods] The activities of alkaline phosphatase(AKP), acid phosphatase(ACP) and esterase(EST) were observed by using calcium cobalt method, lead nitrate method and naphthol method. Results were photomicrographed and analysed quantitatively by image analysis using computer. [Results] The activities of AKP and ACP were decreased while the activities of EST were increased among the infected groups with different infection intensities. [Conclusions] The changes of these enzyme activities might be one of the important mechanisms of action of Lagenidium giganteum against Culex quinquefasciafus larvae.

Objective To construct the suppression subtracted cDNA library of deltamethrin-resistant Aedes albopictus. Methods Total RNA was extracted from the deltamethrin-resistant (R-lab) and -sensitive (S-lab) isolates, mRNA was obtained after purification. Double stranded cDNAs were synthesized after reverse transcription. Two subtractions were performed by suppression subtractive hybridization with S-lab as tester and R-lab as driver or S-lab as driver and R-lab as tester. Enriched different expressed cDNA was cloned into pMD18-T vector to construct subtractive libraries. Results The subtracted cDNA libraries contained 580 and 477 positive clones respectively. The PCR results of 150 clones picked randomly from each library showed that the positive ratio of constructed cDNA libraries was 93%, with a length of cDNA fragments ranged from 150 bp to 750 bp. Conclusion The suppression subtracted cDNA library of deltamethrin-resistant Ae. albopictus is constructed.

Objective To understand the regional distribution and drug resistance of clinical isolates of bacteria flora ,in order to provide the basis for the diagnosis and treatment of bacterial infection .Methods According to the national clinical test proce‐dures operation separation strains ,HX‐21 bacteria identification/susceptibility analyzer bacteria identification and drug sensitive test .Results Among 1 705 strains of isolated bacteria ,Gram‐positive cocci accounted for 39 .8% ,Gram‐negative bacteria accounted for 60 .2% ;separation rate from high to low were:benzene azole resistance westwood 13 .3% coagulase negative staphylococcus , pseudomonas aeruginosa ,12 .0% benzene azole resistance westwood staphylococcus aureus 11 .3% ,did not produce the ultra broad spectrum beta lactamase 10 .7% ,e .coli to produce ultra broad spectrum beta‐lactamase e .coli 9 .9% ,acinetobacter was 7 .1% ,pro‐ducing ultra broad spectrum beta‐lactamase pneumonia klebsiella bacteria 5 .9% ,did not produce the ultra broad spectrum beta‐lac‐tamase pneumonia klebsiella bacteria 5 .7% ,benzene azole westwood sensitive coagulase negative staphylococcus 4 .8% ,enterococ‐cus was 4 .8% ,benzene azole westwood sensitive staphylococcus aureus 4 .5% ,etc .Conclusion The common pathogenic bacteria is gram‐negative bacilli in the common pathogenic bacteria .In negative bacilli infection ,acinetobacter ,pseudomonas aeruginosa ,e .coli , klebsiella pneumoniae ,etc were more common .In staphylococcus aureus strains ,benzene azole westwood drug‐resistant strain ratio is higher than benzene azole westwood sensitive strain rate for 3 times .In addition to the vancomycin and teicoplanin sensitive ,other commonly used antibiotics shows different degrees of resistance .

Objective:To reveal the dynamic characteristics of the intracellular complement mRNA from mouse macrophage ANA-1 treated with LPS or IL-4. Methods:The polarization models of macrophage ANA-1 were established by treating with LPS(1μg/ml) and IL-4(20 ng/ml),respectively. After treating at 3,8,12 and 24 h,the total RNA were abstracted by Trizol lysis methods . The macrophage polarization were estimated by the expression of IL-1β, CCL2 and Arg-1 mRNA detected by Real-time fluorescent quantitative PCR. The intracellular complement C1q, C3, CfB and CRIg mRNA were quantitatively analyzed. Results: The mouse macrophage ANA-1 cells treated with LPS was polarized to M1 since the levels of IL-1β and CCL2 mRNA were up-regulated significantly,in which their 2-△△Ct value were up to 297. 0±31. 0 and 19. 9±3. 3 respectively at 12 h. On the other hand,the ANA-1 cells treated with IL-4 was polarized to M2 because the level of Arg-1 mRNA was obviously higher( the 2-△△Ct value of Arg-1 mRNA was up to 27.3±9.1 at 24 h)(P<0.05).The intracellular complement C1q,C3,CfB and CRIg mRNAs all were up-regulated in different polarized macrophages. The intracellular C1q and C3 mRNA in polarized M2 were significantly higher,in which the peak value of C1q and C3 were to 94. 9±12. 9 and 11. 3±2. 4 at 12 h,respectively(P<0. 05). Reversely,the CfB mRNA in polarized M1 increased obviously,in which its 2-△△Ct was to 61. 4±6. 2 at 12 h. In addition,the CRIg mRNA in both groups was only up-regulated at 24 h,in which the 2-△△Ct value was 6. 5±1. 8 in M1 and 10. 8±3. 2 in M2(P<0. 05). Conclusion: The macrophage ANA-1 cell polarization models were successfully established by treated with LPS or IL-4. The intracellular complement C1q,C3 and CRIg mRNA in polarized M2 were transcripted more than in M1. But the intracellular CfB mRNA in polarized M1 was up-regulated significantly. These results suggested that the dynamic characteristic of complement components in different polarized macrophage would be correlated with its fun-tions.

Objective:To investigate the prognostic value of preoperative inflammatory indicators for hepatocellular carcinoma (HCC).Methods:The retrospective cohort study was conducted. The clinicopathological data of 73 patients with primary HCC who underwent radical partial hepatectomy in the Beijing Tsinghua Changgung Hospital of Tsinghua University from December 2014 to July 2019 were collected. There were 57 males and 16 females, aged from 33 to 81 years, with a median age of 58 years. Results of blood examination indicators at the first time in hospital were determined for patients. Observation indicators: (1) the best cut-off values of?? preoperative inflammatory indicators calculated by the maximally selected rank statistics; (2) follow-up; (3) influencing factors for prognosis of HCC patients; (4) comparison of clinicopathological parameters of HCC patients; (5) comparison of predictive value for overall survival. Follow-up was conducted using outpatient examination and telephone interview to determine postoperative survival of patients up to September 2019. Measurement data with normal distribution were represented as Mean±SD, and comparison between groups was analyzed using the t test. Measurement data with skewed distribution were represented as M (range). The best cut-off values ??for continuous variables were obtained using the maximally selected rank statistics based on survival at endpoint of follow-up. Count data were represented as absolute numbers, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. Kaplan-Meier method was used to calculate survival rates, and Log-rank test was used for survival analysis. Univariate analysis was performed using the Log-rank test. Multivariate analysis was performed using the COX proportional hazard model. The time-dependent receiver operating characteristic curve (ROC) was used to compare the predictive value of independent prognostic factors. Results:(1) The best cut-off values of?? preoperative inflammatory indicators calculated by the maximally selected rank statistics: the best cut-off values of neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR), and prognostic nutrition index (PNI) were 3.46, 131.05, and 45.65. (2) Follow-up: 73 patients were followed up for 31 months (range, 2-57 months). Twenty patients died during the follow-up. (3) Influencing factors for prognosis of HCC patients: results of univariate analysis showed that NLR, PNI, tumor diameter, and tumor differentiation degree were related factors affecting prognosis of patients ( χ2=10.213, 4.434, 5.174, 4.306, P<0.05). Results of multivariate analysis showed that NLR and tumor differentiation degree were independent factors affecting prognosis of patients ( hazzard ratio=4.429, 13.278, 95% confidence interval as 1.662-11.779, 1.056-10.169, P<0.05). (4) Comparison of clinicopathological parameters of HCC patients: of 73 patients, 64 cases had NLR<3.46 and 9 cases had NLR≥3.46. Cases with tumor length >5 cm or ≤5 cm, neutrophils, lymphocytes were 23, 41, (2.9±1.2)×10 9/L, (1.7±0.6)×10 9/L for 64 patients with NLR<3.46, versus 8, 1, (5.8±2.9)×10 9/L, (1.0±0.3)×10 9/L for 9 patients with NLR≥3.46; there were significant differences in above indicators between the two groups ( χ2=7.017, t=2.982, -3.168, P<0.05). (5) Comparison of predictive value for overall survival: time-dependent ROC curves of NLR and tumor differentiation degree for 1-, 2-, 3-, 4-year survival rates had the area under curve of 0.735,0.611, 0.596, 0.574 and 0.554, 0.583, 0.572, 0.556, respectively. NLR had better predictive value for overall survival of patients than tumor differentiation degree. Conclusion:Preoperative NLR is an independent factor affecting prognosis patients, and its predictive efficacy is better than tumor differentiation degree.

Objective: To investigate the characteristics of HIV-1 molecular transmission network among HIV/AIDS patients in parts of Jiangxi Province, so as to provide insights into guiding AIDS prevention and intervention. Methods: The HIV/AIDS patients newly reported from January to June 2018 in Shangrao City, Yichun City and Ganzhou City were recruited, and demographic information and infection routes were collected. Blood samples were obtained to extract HIV RNA, and HIV-1 pol gene was amplified and sequenced using reverse transcription PCR and nested PCR assays. Gene subtypes were analyzed by constructing a phylogenetic tree. Molecular transmission network was constructed using gene-set distance, and the clustering patterns and the characteristics of HIV/AIDS patients within the clusters were analyzed. Results: A total of 305 pol gene sequences were obtained successfully, including 231 males (75.74%), 184 patients aged 50 years and above (60.33%), and 288 patients with heterosexual contact (94.43%). The main subtypes were CRF07_BC and CRF01_AE, accounting for 51.48% and 29.18%, respectively. Ganzhou City had the most genetic subtypes, with 8 types. Under the 1.0% gene distance threshold, 27 molecular clusters were established, with 107 nodes and 150 edges, and the molecular clustering rate was 35.08%. The largest molecular cluster involved 30 patients from 7 counties (districts) of Shangrao City. All of them were CRF07_BC subtypes, had an average age of (63.03±9.46) years, and were infected through heterosexual contact. Among the 17 patients with high transmission risk (degree value≥4), 10 patients came from Yushan County. Conclusions The main subtypes of HIV-1 are CRF07_BC and CRF01_AE in parts of Jiangxi Province, and the subtypes in Ganzhou City are diversified. There may be potential transmission risk points in Yushan County.

Abstract: Objective　This study aims to explore the roles of interferon-stimulated gene cholesterol-25-hydroxylase (CH25H) on the replication of Japanese encephalitis virus (JEV) and its underlying mechanisms, providing potential targets for developing anti-JEV drugs and theoretical support for the research. Methods　First, we investigated whether CH25H could be induced by type Ⅰinterferons or JEV infection in human kidney HEK293T cells through real-time fluorescence quantitative PCR. The cells were stimulated with different concentrations of interferon α (IFN-α), and 12 hours later, CH25H expression was measured via real-time fluorescence quantitative PCR. HEK293T cells were infected with JEV at different multiplicities of infection (MOI), and 24 hours later, changes in CH25H mRNA expression levels were assessed. Eukaryotic expression plasmid pcDNA3.1-CH25H-3×HA was constructed for overexpression of CH25H in HEK293T cells, and its effect on JEV replication was analyzed using real-time fluorescence quantitative PCR and plaque formation assay. The reasonable use concentration of 25-hydroxycholesterol (25HC), the enzyme active product of CH25H, in HEK293T, BHK21, and Vero cells was obtained by CCK8 assay, and the roles of 25-hydroxycholesterol (25HC), the enzyme active product of CH25H, in JEV replication and its impact on JEV life cycle, were further studied by qPCR, viral plaque assay, immunofluorescence assay and adsorption assay. Point mutation was employed to construct an enzymatically inactive mutant, CH25HM, which was transfected into HEK293T cells. After 24 hours of JEV infection, the dependence of CH25H-mediated impact on JEV replication on its enzymatic activity was revealed through qPCR and plaque formation assay. Results　CH25H could be induced by IFN-α in HEK293T cells, but after JEV infection, CH25H mRNA expression was downregulated. Overexpression of CH25H in HEK293T cells significantly inhibited JEV replication. 25HC, the enzyme product of CH25H, can inhibit JEV replication in HEK293T, BHK21, and Vero cells. In terms of mechanism, 25HC can inhibit JEV replication by affecting the virus adsorption process. Expressing the enzymatically inactive mutant CH25HM in HEK293T cells, also significantly inhibited JEV replication. Conclusions　JEV infection down-regulates the expression of interferon-stimulated gene CH25H, while CH25H inhibits JEV replication in a manner dependent on enzyme activity and non-enzyme activity.

The effects of in vivo local expression of recombined human tissue-type plasminogen activator (t-PA) gene on the thrombosis and neointima formation of vein grafts were explored. Jugular vein-to-artery bypass grafting was performed on 72 New Zealand white rabbits. The rabbits were divided into 3 groups according to the different processing methods: transfected t-PA gene group (n = 24), vector group (n = 24) and blank control group (n = 24). Samples of vein grafts were harvested at different time points after surgery. The expression of t-PA gene in vein graft was detected by RT-PCR and the synthesis of t-PA protein by Western-Blot assay. The t-PA activity was measured by chromogenic substrate assay. The Cr51 labeled platelets accumulation in vein grafts was counted. The histopathological changes were compared in intima hyperplasia index among the three groups after operation. The results showed that at the 2nd, 5th, 14th and 28th day after operation, RT-PCR and Western-blot confirmed the expression of t-PA mRNA and protein at the site of gene transfer. The t-PA activity detected on the 2nd, 5th, 14th and 28th day in experimental group was 370.63 +/- 59.44, 344.13 +/- 48.47, 252.87 +/- 51.80 and 161.75 +/- 68.94 U/g respectively, and disappeared on the 60th day and undetected in the control groups. The number of platelets accumulated in the vein grafts in gene group, vector group and blank control group was (85.04 +/- 21.58) 10(6), (225.87 +/- 85.13) 10(6) and (211.7 +/- 78.02) 10(6) respectively. The number of platelets accumulated in gene group was significantly fewer than that in the control groups. Morphometric analysis revealed that intimal hyperplasia was markedly reduced in the t-PA gene group as compared with that in the control groups. It was suggested that the local expression of t-PA gene in vein graft significantly inhibited the accumulation of platelets, thrombosis and concomitant intimal hyperplasia, by which stenosis of bypass graft could be prevented effectively.

Objective To observe the surface electromyographic characteristics of the bilateral submen-tal muscles in dysphagia secondary to unilateral brainstem stroke. Methods A total of 25 subjects were recrui-ted. There were 8 stroke patients with dysphagia secondary to a left brainstem stroke and 7 stroke patients with dysphagia secondary to a right brainstem stroke. There were also 10 healthy controls matched in age and gender. The duration and peak amplitude of the submental muscle when swallowing 5 ml of warm water were recorded u-sing a surface electromyograph. Results The average amplitude of the left submental muscle in patients with a left brainstem stroke was significantly longer than that of those with a right brainstem stroke, but no significant differences in average duration were observed. Conversely, the amplitude of the right submental muscle in pa-tients with a right brainstem stroke was significantly longer than that of those with left brainstem stroke, but again there were no significant differences in duration. No significant differences were observed among the healthy con-trols. The amplitude and duration of both the affected and healthy sides of the patients were of course significantly longer or stronger than those of the healthy controls. Conclusion The swallowing function of the bilateral sub-mental muscles may be impaired among unilateral stroke survivors with dysphagia. The damage on the affected side is more severe than on the opposite side.

BACKGROUND:Early detection and accurate staging diagnosis of heart failure are the basis of good clinical therapy efficacy. Due to lack of simple and effective staging model for the diagnosis of heart failure, it is difficult to diagnose heart failure in clinics, leading to poor control of heart failure. OBJECTIVE:To establish the disease staging model based on Adaboost and SVM for heart failure, and improve the accuracy of diagnosis and staging of heart failure. METHODS:A total of 194 cases were roled into this study, including heart failure patients and healthy physical examination persons. According to the stage standards formulated by American Colege of Cardiology and American Heart Association, specific clinical feature parameters closely related to heart failure were colected and selected. Based on clinical diagnosis results and using Adaboost model and SVM model, we trained the models for heart failure diagnosis and staging, thus obtaining diagnosis model. RESULTS AND CONCLUSION: The parameters included stroke volume, cardiac output, left ventricular ejection fraction, left atrial diameter, left ventricular internal diameter at end-systole, N-terminal pro-brain natriuretic peptide and heart rate variability. As for the Adaboost model, its sensitivity and specificity was 100% and 94.4%, respectively. At the same time the SVM model had good sensitivity and specificity, 86.5% and 89.4% respectively. Adaboost classification model can be accurate in the diagnosis of heart failure symptoms, the accuracy reached 89.36%. On the basis of the diagnosis of heart failure, the SVM classification model is effective in staging the severity of heart failure, staging accuracy for staging B and C was 86.49% and 81.48%, respectively. The findings indicate that, combining Adaboost and SVM machine learning models could provide an accurate diagnosis and staging model for heart failure.