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.

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.

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.

OBJECTIVETo investigate the effects and mechanism of Tripterygium hypoglaucum (THH) on serum IL-1, IL-6, and TNF-alpha in chronic, nephritis rats.

METHODThe rabbit serum of anti-rat kidney was initially prepared, and then injected into normal rats to induce the formation of chronic glomerulonephritis. In this model, THH was administrated for 4 weeks, while saline and prednisone were respectively used as negative and positive controls. Some of laboratory parameters were observed from the rats above.

RESULTTHH not only significantly decreased urine protein, reduced serum urea nitrogen, but also decreased the releases of inflammatory mediators (such as IL-1, IL-6 and TNF-alpha).

CONCLUSIONTHH is effective in treating rat nephrotoxic serum glormerulonephritis, its mechanism probably related to decreasing inflammatory mediator levels.

Animals ; Chronic Disease ; drug therapy ; Female ; Interleukin-1 ; blood ; Interleukin-6 ; blood ; Kidney ; drug effects ; pathology ; Male ; Nephritis ; blood ; drug therapy ; pathology ; Plant Extracts ; pharmacology ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Time Factors ; Tripterygium ; chemistry ; Tumor Necrosis Factor-alpha ; blood

Objective To clone ORF of DcR3 gene and insert it into eukaryotic expression vector and express it in COS-7 cells. Methods Encoding sequence of human DcR3 gene was cloned by PCR and sequenced. The sequenced ORF was cloned into eukaryotic expression vector pAAV-IRES-hrGFP to construct recombinant plasmid. COS-7 cells were transfected with recombinant plasmid by lipofectamine2000. Expression of recombinant DcR3 gene was verified by Western blotting and confocal microscopy. Results A 1 000-bp gene segment was obtained by PCR and inserted into pAAV-IRES-hrGFP to construct recombinant plasmid. The gene segment was proved to be encoding sequence of human DcR3 gene by sequencing. DcR3 expression in COS-7 cells was verified by Western blotting and confocal microscopy. Conclusion DcR3 gene was successfully cloned and expressed in COS-7 cells.

Objective:To establish a UVB damage cell model with HaCaT cells to investigate the protective effects of Zinc sulfate on the cell damage caused by UVB and its relevant mechanisms .Methods: The cells were divided into normal group , Zinc group,UVB group,Znic and UVB group.The addition of Zinc sulfate to the HaCaT cells was conducted 24h prior to the irradiation to the cells by UVB.Cell apoptosis was detected by Western blot and the expression of metallothionein and NF -κB/p65 were measured by im-munohistochemistry.Results:Compared with normal and Zn+UVB group, Bax/Bcl-2 rate in UVB group increased.Compared with normal group ,MT expression levels in UVB group ,Zn group increased ,and compared with UVB group ,MT expression level in Zn+UVB group increased .Compared with normal group and Zn+UVB group,NF-κB/p65 expression level in UVB group increased .Conclusion:Zinc sulfate alleviates the apoptosis of HaCaT cell induced by UVB because of the expression of MT .