OBJECTIVE To investigate whether efflux mechanism is involved in fluoroquinolone-resistant Klebsiella pneumoniae strains isolated in China. METHODS We compared the ciprofloxacin accumulation in clinically isolated K. pneumoniae with or without CCCP by fluorospectrophotometry. Use reverse transcriptase-polymerase chain reaction (RT-PCR) to measure the mRNA expression level of acrAB-tolC efflux gene. RESULTS The accumulation of ciprofloxacin in resistant strains was lower than that in susceptible ones, and it could increase to a high level nearly to the susceptible strains. The mRNA level of efflux gene acrA was higher in resistant strains than in susceptible ones. CONCLUSIONS Efflux mechanism is associated with the resistance to fluroquinolones in K. pneumoniae strains isolated in China and CCCP can inhibit its active efflux mechanism , which provides a sensitive method to detect the active efflux system of K. pneumoniae.

Objective To investigate the effect of S100A4 silencing on tumor related gene COX-2,bcl-2,Surviving,MMP-9 mRNA expressions of pancreatic cancer BxPC-3,AsPC-1 cells,and explore their relationship.Methods Small interfering RNA interfering S100A4 gene (siRNA-S100A4) was applied to transfect human pancreatic cancer BxPC-3,AsPC-1 cells,and nonhomologous siRNA-C was used as negative control,and cells without transfection were used as control group.The expressions of S100A4,COX-2,Survivin,MMP-9,bcl-2 mRNA after interference were detected by using RT-PCR.Results S100A mRNA expressions of BxPC-3's control group,siRNA-C group,siRNA-S100A4 group were 0.661 ± 0.023,0.659 ± 0.043,0.379 ± 0.039,and expressions of COX-2 mRNA were 0.760 ± 0.026,0.830 ± 0.017,0.443 ±0.006,and expressions of Survivin mRNA were 0.948 ± 0.049,0.909± 0.081,0.068 ± 0.006,and expressions of bcl-2 mRNA were 0.462 ±0.018,0.421 ±0.049,0.184 ±0.025,and expressions of MMP-9 mRNA were 0.813 ± 0.008,0.908 ± 0.063,0.246 ± 0.027.S100A mRNA expressions of AsPC-I's control group,siRNA-C group,siRNA-S100A4 group were 0.641 ± 0.042,0.626-± 0.053,0.320 ± 0.081,and expressions of COX-2 mRNA were 0.727 ± 0.021,0.743 ± 0.025,0.560 ± 0.035,and expressions of Survivin mRNA were 0.994 ± 0.032,0.984 ± 0.049,0.063 ± 0.005,and expressions of bcl-2 mRNA were 0.458 ±0.004,0.537 ± 0.046,0.181 ± 0.007; and expressions of MMP-9 mRNA were 0.698 ± 0.011,0.718 ± 0.073,0.199± 0.013.The expressions of S100A,COX-2,Survivin,bcl-2,MMP-9 mRNA in groups with siRNA-S100A4 transfection were significantly lower than those of siRNA-C group and control group (P ＜0.01),but the difference between siRNA-C group and control group was not statistically significant.Conclusions S100A4 plays a role in the pathogenesis of pancreatic cancer through up-regulation of COX-2,Survivin,bcl-2,MMP-9 expressions.

Objective: To express vascular basement membrane-derived multiple peptide (VBMDMP) in Pichia pastoris and to identify its bioactivity. Methods: PCR technique was used to obtain the target fragment GST-VBMDMP, which was then cloned into pPIC9K vector. The resultant vector pPIC9K-GST-VBMDMP was transfected into Pichia pastoris cells with spheroplast and the positive recons was identified. The His + Muts transformant was shake-cultured in MGY at 30 ℃. The culture supernatant (1 ml) was collected for preservation at-70 ℃ every 24 hours while maintaining the concentration of methanol at 0.5% in the culture medium. SDS-PAGE analysis was used to detect the protein expression after 8 days and then animal and cell experiments were performed with GST-VBMDMP. Results: SDS-PAGE analysis found that protein express increased during 1-7 days and decreased after 8 days in MGY medium; GST-VBMDMP fused protein was obtained by Glutathione Sepharose 4B and it inhibited the artery endothelial cell tube structure formation in C57BL/6 mouse; GST-VBMDMP( 2, 6, 10 mg/kg)also significantly inhibited the primary cancer Lewis mouse (tumor inhibitor rates being 96.6%, 82.1%, and 61.2%, respectively)and metastatic lung tumors(tumor inhibitor rates being 96.8 %, 87.9 %, and 75.3%, respectively)(P

Epigenetics is the important component of functional genomics. It can be defined as the study of the interplay between environment and genetics, and the study of the heritable change that is not strictly dependent on DNA sequence. Nasopharyngeal carcinoma (NPC) is the common malignant tumor in south China. As a polygenetic inheritance tumor, NPC has the characteristic of obvious tendency of familial aggregation, possesses genomic instability, and is vulnerable to physical, chemical and biological carcinogenic factors. This specific etiological system of NPC refers that it is one of the best model for studying cancer epigenetics. The main point of this review focuses on the progress of studying on the effects of DNA methylation, histone modification, chromatin remodeling and non-coding RNA regulation on the pathologenesis of NPC, and suggests future directions to thoroughly explore the epigenetic pathologenesis of NPC. Moreover, it will open a new prerequisite foundation for finding the epigenetic molecular marks for screening the high-risk susceptibility population, early diagnosis, treatment and prognostic indentification of NPC.

Objective To evaluate the clinical significance of implanting an inferior vena cava filter to prevent pulmonary embolism.Methods Reviewed the causality of 180 cases,which was undertaken the ultrasound examination,patients were diagnosed as the deep vein thrombosis,male patients were 128 cases, female patients were 52 cases,median age was 65 years old,all received the transfemora] implantation of inferior vena cava filter implantation.Results All the operations were successfully conducted,filters were placed into inferior vena cava at 1-3 cm inferior of renal vein,without hematoma in the puncture position and thrombosis.One hundred and forty-nine cases (82.8%) were followed up at 1,6,12 months respectively, 55 cases(30.6%)were followed up more than 36 months, no filter drift,deformation and inferior vena perforation complications was observed.There was no fatal pulmonary embolism occurred for all the cases,the inferior vena unobstructed rate was 95.6%.Conclusion Vena cava filter is an effective and safe method which can prevent and cure pulmonary embolism.

Objective To explore the effects of L-carnitine on the plasma lipid profile and liver function in elderly patients receiving total parenteral nutrition after abdominal operations.Methods In this prospective blinded randomized controlled trial,24 eligible elderly patients were given 6-day total parenteral nutrition.They were further equally divided into L-carnitine group(administered with L-carnitine 50 mg/kg)and control group(without L-carnitine).The changes of lipid profile and liver function and the clinical outcomes were recorded and compared.Results The plasma triglyeride levels were lower in L-carnitine group than in control group on the 4th and 7th post-operative day,but there were not significant difference(P>0.05);There was a significant difference between the two groups in the change of the 4th post-operative day and pre-operative day(P<0.05).However,the liver function indicators were not significantly different between these two groups.Conclusion Administration of L-carnitine during total parenteral nutrition can improve the triglyeride metabolism in elderly patients after abdominal operations and may benefit the recovery of liver function.

Objective To investigate the clinical effects of percutaneous trasluminal angioplasty (PTA) for ischemic lesions of below-the-knee(BTK) arteries in elderly patients.Methods Clinical data of 56 elderly patients(the PTA group) with ischemic lesions of BTK arteries who had received PTA treatment were retrospectively analyzed.The ankle brachial index (ABI) and the skin temperature of the affected foot were recorded and analyzed before and after treatment.The clinical effects were compared with those of conservative treatment 31 patients(the control group) had received during the same period.Results PTA was successfully performed in all of the 56 cases.The skin temperature rose,claudication disappeared or the claudication distance increased,rest pain disappeared or lessened,and no severe complications or death occurred in the PTA group.ABI increased from (0.41±0.12) before treatment to(0.63±0.21),(0.65±0.19),(0.67± 0.22),(0.61±0.20),and (0.58±0.18) at 24 hours,1,3,6 and 12 months after treatment,respectively.The skin temperature increased from(28.4±2.1)℃ before treatment to(35.4±2.4)℃,(35.6±2.1)℃,(35.8±2.4)℃,(34.6±2.1)℃ and(34.3±2)℃ at 24 hours,1,3,6 and 12 months after treatment,respectively.There were statistically significant differences in ABI and the skin temperature between before treatment and any time point after treatment(all P＜0.05) and between the PTA group and the control group at any time point after treatment(all P＜0.05).Conclusions PTA is a safe and effective treatment for ischemic lesions of BTK arteries in elderly patients.

Objective To test the expression level of IL-37 in peripheral blood mononuclear cells (PBMCs)of patients with primary biliary cirrhosis (PBC)and further explore its clinical significance in the pathological process of PBC.Methods Pe-ripheral blood samples were collected from 42 patients diagnosed as PBC and 38 health individuals examined at the same time during June 2013 to August 2015 in Changhai Hospital.PBMCs were separated by sucrose density gradient centrifugation, qualified Real Time-Polymerase Chain Reaction (qRT-PCR)was used to measure IL-37 mRNA expression level in PBMCs. Enzyme-Linked Immuno Sorbent Assay (ELISA)was to measure the protein level of IL-37,IL-6,IL-17,TNF-α,TGF-β,IL-18 and IL-23 in plasma.Meanwhile,the pathological stages of PBC cases were recorded.Pearson correlation analysis was performed on IL-37 and IL-6,TNF-α,IL-17,TGF-β,IL-18 and IL-23.Spearman rank correlation analysis was on IL-37 and pathological stages of PBC.Results The mRNA and protein level of IL-37 in experimental and controlled group were 2.81 ±0.94 vs 1.09±0.56,356.14±169.36 pg/ml vs 86.68±48.23 pg/ml separately(t=9.811,9.462,P<0.000 1),with sta-tistical differences.The correlation analysis showed that IL-37 was positively related with IL-17,TNF-α,IL-6 and TGF-β(r=0.561 2,0.661 9,0.672 1,0.765 3,P<0.001),and disease stages (Ⅰ~Ⅳ)(rs=0.348 9,P<0.05).Conclusion IL-37 might involve in the pathological process of PBC,and it is significant for disease prediction and diagnosis.

Objective To explore the effects of microRNA(miRNA)- 30c knockdown on proliferation,diffe-rentiation of P19 cells. Methods miRNA - 30c knockdown plasmid(miRNA - 30c knockdown group)or no - load vector(negative control group)was transfected into P19 cells by lipo2000 and stable cell lines were selected by Blastici-din;Dual luciferase reporter gene system was used to confirm miRNA - 30c knockdown. Cell counting kit - 8(CCK - 8) assay was adopted to detect cell proliferation activity. An inverted microscope was used to observe morphological chan-ges of P19 cell differentiation. Cells were induced to differentiated to myocardiocyte with dimethyl sulfoxide(DMSO). Differentiation marker genes including cTnT,NKX2. 5,GATA4 relative mRNA expression levels were detected with real - time quantitative polymerase chain reaction,respectively. Results Observation of green fluorescent protein ex-pression under a fluorescence microscope indicated similar transfection efficiencies,and miRNA - 30c knockdown re-leased the activity of target gene Gli2. As a result,miRNA - 30c knockdown vector was constructed successfully(P ﹤0. 001). During differentiation of mouse P19 cells into myocardial cells,the beating cell clusters in miRNA - 30c knockdown cells were much lower than those in the control cells,and cTnT,NKX2. 5,GATA4 in miRNA - 30c knock-down cells showed significantly lower expression than those in the control cells( all P ﹤ 0. 05). Conclusions miRNA - 30c inhibits the P19 cell proliferation and differentiation. This study gives us a new insight of heart develop-ment and we need more efforts on exploring the deep function of heart diseases.

Objective:To evaluate the efficacy of quantitative fecal immunochemical test (fecal immunochemical test, FIT) in the screening of colorectal cancer and precancerous lesions.Methods:47 243 patients who underwent quantitative FIT screening for colorectal cancer in the Health Management Department of Shenzhen People′s Hospital from January 2019 to October 2020 were enrolled as subjects. Colonoscopy was recommended for patients with positive quantitative FIT. A follow-up was done after one year to compare the results of the quantitative FIT positive group and the negative group after colonoscopy. Data were adjusted by propensity score matching method and a receiver operating characteristic curve ( ROC) was established to evaluate the diagnostic effect of quantitative FIT combined with colonoscopy on colorectal cancer and precancerous lesions. Results:A hemoglobin concentration>100 μg/L was set as a positive threshold. There were 2 472 positive cases of quantitative FIT, and the positive rate was 5.23%. After one-year follow-up, 284 patients had completed colonoscopy; the colonoscopy compliance was 11.49%. Of the negative population, 1 493 patients selected colonoscopy within one year. Compared with the results of gold standard colonoscopy, the sensitivity and specificity of quantitative FIT for screening for advanced adenoma and cancer were 26.53% and 86.54%, respectively. The sensitivity for colorectal cancer screening was 94.44% and screening for advanced adenoma was 22.77%. After propensity score matching, there were 256 FIT positive patients and 705 FIT negative patients. Colorectal cancer was the outcome variable, while FIT combined with colonoscopy, age combined with colonoscopy, and FIT combined with age combined with colonoscopy were the diagnostic indicators. The areas under the curve were 0.841(95% CI:0.778-0.904), 0.677(95% CI: 0.535-0.820), and 0.882(95% CI:0.807-0.958), respectively. Conclusion:Quantitative FIT has a high susceptibility to opportunistic screening for colorectal cancer, and a low sensitivity to advanced adenoma. At the same time, quantitative FIT combined with colonoscopy has a good diagnostic accuracy for colorectal cancer, which is better than the effect of using age as a cut-off point for colonoscopy. Quantitative FIT combined with age and colonoscopy has the best effect on screening for colorectal cancer. Quantitative FIT has a high sensitivity to colorectal cancer and a low sensitivity to advanced adenoma.