ObjectiveTo investigate the expression and the clinical significance of diacylglycerol kinase α (DGKα) and protein kinase C (PKC) in human hepatocellular carcinoma (HCC).MethodsDGKα and PKC expressions in the samples from 60 pathologically confirmed HCC patients were analyzed by immunohistochemistry. The relationship between DGKo expression and clinical pathology factors was analyzed.ResultsThe expression positive rates of DGKo and PKC were highest in normal liver tissues [90.0％ (9/10) and 100.0％ (10/10)].The positive rates were 81.7 ％ (49/60) and 71.7 ％ (43/60) in HCC tissues,respectively,and were 58.3 ％ (35/60) and 61.7 ％ (37/60) in carcinoma adjacent tissues,respectively.In three liver tissues,the positive rates of DGKα and PKC were significantly different (P ＜0.05).The location of both kinases in hepatocytes translocated from cytoplasm/nucleus to membrane.The expressions of DGKα and PKC were positively correlation(r =0.495, P ＜ 0.05), The DGKα expression was correlated to differentiation type,portal venous tumor thrombus and TNM staging(all P ＜ 0.05).ConclusionDGKa is expressed with high activity in advanced stage and poorly differentiated HCC. It may be promote the pathological process of HCC.

Rhamnolipid biosurfactant is mainly produced by Pseudomonas aeruginosa that is the opportunistic pathogenic strain and not suitable for future industrial development. In order to develop a relatively safe microbial strain for the production of rhamnolipid biosurfactant, we constructed engineered Escherichia coli strains for rhamnolipid production by expressing different copy numbers of rhamnosyltransferase (rhlAB) gene with the constitutive synthetic promoters of different strengths in E. coli ATCC 8739. We further studied the combinatorial regulation of rhlAB gene and rhaBDAC gene cluster for dTDP-1-rhamnose biosynthesis with different synthetic promoters, and obtained the best engineered strain-E. coli TIB-RAB226. Through the optimization of culture temperature, the titer of rhamnolipd reached 124.3 mg/L, 1.17 fold higher than that under the original condition. Fed-batch fermentation further improved the production of rhamnolipid and the titer reached the highest 209.2 mg/L within 12 h. High performance liquid chromatography-mass spectrometry (LC-MS) analysis showed that there are total 5 mono-rhamnolipid congeners with different nuclear mass ratio and relative abundance. This study laid foundation for heterologous biosynthesis of rhanomilipd.
Bacterial Proteins ; genetics ; Batch Cell Culture Techniques ; Decanoates ; Escherichia coli ; metabolism ; Fermentation ; Glycolipids ; biosynthesis ; Hexosyltransferases ; genetics ; Industrial Microbiology ; methods ; Multigene Family ; Promoter Regions, Genetic ; Pseudomonas aeruginosa ; Rhamnose ; analogs & derivatives ; biosynthesis ; Surface-Active Agents ; metabolism

Objectives It was constructed that the replication defective adenoviral vectors carried the short hairpin sequences of mouse SCP2.And we will make a further study of mechanism between SCP2 gene and cholesterol stone in gallbladder.Methods The short hairpin sequences of mouse SCP2 were cloned by two-step PCR,and connected together with the plasmid pDC312.The Admax Adenoviral Vector System was used to generate the replication defective adenoviral vectors,which were purified by CsCl method.The processes of TCID50 were applied to detect the titers of the adenoviral vectors.Furthermore,Protein levels of SCP2 were determined by Western blot analysis,and the levels of SCP2、CYP7A1、HMGCR mRNA from the hepa1-6 cell of mouse were measured by the usage of RT-PCR.Results SCP2mRNA and SCP2 protein were down-regulated by the replication defective adenoviral vectors carried the SCP2-shRNA.With the decreasing SCP2mRNA,the levels of HMGCRmRNA were down-regulated at same the time,while CYP7A1mRNA were up-regulated.Conclusions The replication defective adenoviral vectors carried SCP2-shRNA were constructed successfully.The lower levels of SCP2 could affect the activities of HMG-CoA reductase and CYP7-a enzyme,which caused the variations of cholesterol metabolism and then decreased the formation of cholesterol stone.

To develop a standard defibrillation energy source which can output monophasic and biphasic de-fibrillation impulses and can display standard energy values. The storage capacitor was charged by single-end flyback transformator and discharged through the H bridge composed of insulated gate bipolar transistor (IGBT). Impulse voltage and current were collected by divider while discharging, and the energy value was calculated by STM32. The en-ergy value and waveform were displayed through the control module. The energy source could output both monophsic and biphasic waveforms, and the accuracy of displayed value was higher than ±2% or ±1 J. The standard defibrillation energy source can be used as standard device for defibrillator analyzer, and the metrological trace-ability system of defibrillation energy may come to be completed.

Objective To evaluate the clinical diagnostic application and operative efficacy of the expression of NKG2D in peripheral blood CD+8 NKT cell and its ligand sMICA in patients with esophageal or cardiac carcinoma.Methods The peripheral blood NKG2D positive CD+8 NKT cell percentage was concomitantly determined by flow cytometry in 53 preoperative patients including 29 postoperative patients with esophageal or cardiac carcinoma and 30 healthy controls.The serum sMICA was determined by ELISA.Results The peripheral blood NKG2D positive CD+8 NKT cell percentage in patients was significantly lower than that in controls [(77.632±8.972) % vs (89.053±6.515) %] (t = -6.113,P ＜0.05); with stage Ⅱ,Ⅲ,Ⅳ,it decreased significantly in order (F = 99.251,P ＜0.01);with lymph node metastasis lower than that without lymph node metastasis (t = -10.384,P ＜0.01); squamous carcinoma was higher than adenocarcinoma (t =9.899,P ＜0.01); postoperative was significantly higher than preoperative (t =-4.319,P ＜0.01).The level of serum sMICA in patients was significantly higher than that in controls [(326.28±85.407) pg/ml vs (210.00±92.560) pg/ml](t =7.292,P ＜0.01); with stage Ⅱ,Ⅲ,Ⅳ,it increased significautly in order (F =63.355,P ＜0.01); with lymph node metastasis higher than that without lymph node metastasis (t =7.770,P ＜0.01); squamous carcinoma was lower than adenocarcinoma (t =-7.593,P＜0.01); postoperative was significantly lower than preoperative (t =7.027,P ＜0.01).Serum sMICA could inhibit peripheral blood CD+8 NKT cell activation receptor NKG2D (F =142.773,P ＜0.05),determination coefficient R2 = 0.7368.Conclusion The level of peripheral blood CD+8NKT cell activation receptor NKG2D and serum sMICA in patients could be an assistant indicator for

Objective To construct recombinant adeno-associated virus and lentivirus carrying siRNA of TIMP-1 and to investigate the efficiency of infection and short-term inhibitory effect of TIMP-1 gene expres-sion on rat hepatic stellate cells. Methods One pair of siRNA which could effectively inhibit expression of the TIMP-1 gene in HSC-T6 was screened and cloned into AAV vector and lentiviral vector to construct the recombinant AAV/siRNA-TIMP-1/GFP and Lenti/siRNA-TIMP-1/GFP. AAV/GFP and Lenti/GFP as neg-ative control were also obtained. Experiments were assigned to five groups: AAV/siRNA-TIMP-1/GFP, AAV/GFP, Lenti/siRNA-TIMP-1/GFP, Lenti/GFP group and mock group. Rat HSC-T6 cells were infected by these recombinant viruses at a concentration of MOI by 10. To monitor the efficiency of infection, fluores-cence microscope and flow cytometer were used. After 7 d post-infection, Western blot was used to detect the TIMP-1 protein expression. Results HSC-T6 had no significant changes after infection. The efficiency of infection in AAV/GFP and Lenti/GFP group were 72.7% and 70.0%, AAV/siRNA-TIMP-1/GFP and Lenti/siRNA-TIMP-1/GFP group were 64.58% and 61.86%. The protein expression levels of TIMP-1 in HSC-T6 cells at 7 d post-infection by the recombinant AAV and Lentivirus were decreased 40.0% compared with those in mock control and normal HSC-T6 (P<0.05). Conclusion Recombinant AAV/siRNA-TIMP-1/GFP and Lenti/siRNA-TIMP-1/GFP could effectively infect HSC-T6 with similar efficiency and suppress the expression of TIMP-1 in rat HSC-T6 remarkably.

Objective To investigate the epidemiological characteristics of renal transplantation recipients, effective prevention and control measures. Methods A total of 456 renal transplant recipients were monitored from January 2014 to December 2017. Postoperative infection including baseline data, infection site and infectious pathogen type was analyzed. Results Among 456 renal transplant recipients, 78 cases (17.1%) developed nosocomial infection. Postoperative infection time was 9(3-21) d. Infection sites mainly included the lower respiratory tract, urinary system and blood infection. Infection pathogens consisted of Staphylococci (n=13), Enterococcus faecium (n=6), fungi (n=6), Stenotrophomonas maltophilia (n=4), Acinetobacter baumannii (n=4), Pseudomonas aeruginosa (n=4), Staphylococcus epidermidis (n=4), Klebsiella pneumoniae (n=1), Escherichia coli (n=1) and other negative bacteria (n=9). Among them, 11 cases (14%) were infected with multi-drug resistant bacteria, and 4 cases died. Conclusions In renal transplant recipients, the incidence of nosocomial infection is relatively high, with early postoperative onset, common multiple drug-resistant bacterial infection and high mortality. Preoperative preparations should be fully implemented, postoperative lower respiratory tract infection should be actively prevented and prevention and treatment measures for multidrug-resistant bacteria should be standardized.

Objective:To investigate the Effect of madecassoside (MC) on hippocampal NR2B expression and cognitive function following traumatic brain injury (TBI).Methods:One hundred and sixteen SD rats were provided by the experimental animal center of Medical College in Xi′an Jiao Tong University. Rats were randomly divided into four groups ( n=29) as following: Control group (Con group), TBI group, MC treated TBI rats group (TBI+ MC group) and MC treated control rats group (Con+ MC group). NR2B protein levels in hippocampus were detected by Western-blot at 12 h, 1 d, 3 d, 7 d, 14 d and 21 d post trauma. Hippocampal NR2B positive cells were studied by immunohistochemical (IHC) staining at 21 d post trauma. Cognitive functions of rats were evaluated by Morris water maze (MWM) test from 21 d to 25 d post trauma. Results:Expression of hippocampal NR2B in TBI group at 12 h, 1 d and 3 d post injury were 3.31±0.28, 2.17±0.31 and 1.96±0.31, which presented statistical difference among the three time-points ( P<0.05) and were significantly increased compared to Con group ( P<0.05). However, there was no difference of NR2B level between TBI group (0.93±0.13) and Con group ( P>0.05) at 7 d post injury. In addition, NR2B expression in TBI group at 14 d and 21 d post injury were 0.45±0.04 and 0.21±0.04, which were much lower than that in Con group. IHC staining revealed that the number of hippocampal NR2B positive cells in TBI group (33.26±9.71) were lesser than that of Con group (86.62±17.05). Increased Escape latency, decreased platform crossing times and target quadrant duration were found in TBI group compared with Con group ( P<0.05). Hippocampal NR2B expression in TBI+ MC group at 12 h, 1 d and 3 d post injury were 2.37±0.34, 1.38±0.22 and 1.14±0.16. Difference among the three time-points exhibited statistical significance ( P<0.05). In addition, they were much lower than that in TBI group at the same time-points ( P<0.05), but no difference was found between TBI+ MC group (0.97±0.06) and TBI group at 7 d post injury ( P>0.05). Moreover, NR2B expression in TBI+ MC group at 14 d and 21 d post injury were 0.86±0.08 and 0.52±0.06, which were much higher than that in TBI group ( P<0.05). IHC staining showed the number of hippocampal NR2B positive cells in TBI+ MC group (69.08±12.24) were much more than that of TBI group (33.26±9.71). A decrease of escape latency, an increase of platform crossing times and target quadrant duration were found in TBI+ MC group compared to TBI group ( P<0.05). Conclusion:MC treatment could inhibit the up-regulation of NR2B in hippocampus at early period of TBI and prevent the down-regulation of NR2B at advanced stage of TBI, which led to a remarkable improvement for the cognitive dysfunction caused by TBI.

Objective:To explore the predictive effect of CHA2DS2-VASc score on the long-term prognosis of acute pulmonary embolism (APE).Methods:Patients who were diagnosed with acute pulmonary embolism in the department of respiratory medicine of Binzhou Second People′s Hospital and Hebei Provincial People′s Hospital from October 2014 to October 2018 were continuously included, and the included patients were divided into two groups according to the CHA2DS2-VASc score: 319 cases in the low CHA2DS2-VASc group (<4 points), and 79 cases in the high CHA2DS2-VASc group (≥4 points). Then the propensity score matching method was used to balance the covariates between the two groups, and then the CHA2DS2-VASc score was used to predict the long-term prognosis of acute pulmonary embolism.Results:The Geneva score, D-dimer level and APE-related adverse events in high CHA2DS2-VASc group were significantly higher than those with low CHA2DS2-VASc group, with statistically significant differences ( P<0.05). The multiple COX regression model showed that the incidence of pulmonary embolism associated adverse events was significantly increased 2.820-fold (95% CI: 1.366-5.822) in the high CHA2DS2-VASc group compared with in the low CHA2DS2-VASc group. After propensity score matching, high CHA2DS2-VASc score was still a strong predictor of poor prognosis in patients with acute pulmonary embolism ( HR: 3.421, 95% CI: 2.164-5.408). Conclusions:After using propensity score matching method balances confounding bias, high CHA2DS2-VASc score is still an independent prognostic risk factors of acute pulmonary embolism.

Objective To explore the effect of chloroquine on death receptor 5 (DR5) expression of hepatocellular carcinoma Huh7 cells and cell proliferation and apoptosis induced by tumor necrosis factor related apoptosis-inducing ligand (TRAIL).Methods Huh7 cells were divided into four groups:the control group (1∶1 000 dimethyl sulfoxide),TRAIL group (50 μg/L),chloroquine group (10 μmol/L) and TRAIL +chloroquine group (TRAIL 50 μg/L + chloroquine 10 μmol/L).Thiazolyl blue tetrazolium bromide (MTT) assay was used to determine the proliferation activity of cells,immunofluorescence was used to detect the expression of DR5,4',6-diamidino-2-phenylindole (DAPI) staining was used to observe cell apoptosis and Western blot was used to detect the expression of cleaved poly ADP-ribose polymerase (PARP).Results TRAIL treatment could decrease Huh7 cells proliferation activity;when compared with the cell viability in the control group,the cell proliferation inhibition rate of chloroquine group,TRAIL group and TRAIL+ chloroquine group was (89±8) ％,(53±10) ％ and (27±7) ％,respectively;compared with TRAIL group alone,cell proliferation activity was decreased in TRAIL+ chloroquine group (t =3.922,P =0.017).The expression of DR5 was upregulated in chloroquine group,and the cell apoptosis signaling was activated in TRAIL + chloroquine group.The cell apoptosis rate of TRAIL group and TRAIL + chloroquine group was (10.0±2.3) ％ and (20.4±4.0) ％,respectively,and there was a statistical difference (t =3.894,P =0.018).Conclusion Chloroquine can enhance the cell chemosensitivity to TRAIL treatment by upregulating the expression of DR5 in Huh7 cells.