Objective To investigate the effects of special siRNA targeting TLR4 gene (TLR4-siRNA) on hyperoxia-induced secretion of inflammatory factors from human alveolar epithelial cell line A549.Methods Human alveolar epithelial cell line A549 was purchased from Chongqing Children Hospital and cultured in six-well tissue culture plates and randomly divided into 4 gorups:A549 cells + ambient air group (group C) ; A549 cell + hyperoxia group (group H); A549 cells transfected with TLR4-siRNA + hyperoxia group (group TR) and A549 cells transfected with control-siRNA + hyperoxia group (group NR).TLR4-siRNA and control-siRNA were transfected into A549 cells through lipofectamine 2000.The efficiency of transfection was assessed by detection of FAM expression using flow cytometry.The cells were exposed to 95％ O2-5％ CO2 delivered at 1 L/min for 30 min in groups H,TR and NR.TLR4 mRNA and protein expression was detected by RT-PCR and cytometry respectively.The NF-κB activity in A549 cells was measured by EMSA and the IL-6 and IL-8 levels in supernatant were determined by ELISA.Restults Hyperoxia significantly up-regulated TLR4 mRNA and protein expression and increased NF-κB activity in cells and IL-8 and IL-6 concentrations in supematant in group H as compared with group C.Transfection with TLR4-siRNA significantly attenuated hyperoxia-induced up-regulation of TLR4 mRNA and protein expression and increase in NF-κB activity in the cells and IL-6 and IL-8 concentrations in supernatant in group TR as compared with group H,but transfection with control-siRNA did not.Conclusion TLR4-siRNA could significanly inhibit hyperoxia-induced secretion of inflammatory factors from human alveolar epithelial cell line A549.

Objective To study the effect of Shenfu injection combined with low dose glucocorticoid on endothelial function and blood coagulation in elderly patients with severe pneumonia. Methods 120 patients with severe pneumonia in our hospital from January 2014 to January 2017 were selected,and randomly divided into treatment group and control group,60 cases in each groups.. All patients were treated with conventional treatment. On this basis, the control group was treated with small dose of glucocorticoid. The treatment group was treated with Shenfu Injection on the basis of the control group. The severity, endothelial function and coagulation function of the two groups were compared at different times after treatment. Results The scores of APACHE and 120 h in the two groups were significantly lower than those in the control group (P<0.05). The score of the treatment group was lower (P<0.05) than that of the control group (96 h). After treatment, the two groups of patients with s E-selectin, V WF, APTT, D-D were significantly decreased, PLT significantly increased, significant difference (P<0.05); compared with the control group, s E-selectin, V WF, APTT, D-D and other indicators of the treatment group decreased and PLT increased even more sharply, there is statistical significance (P<0.05). Conclusion Shenfu injection combined with small dose of glucocorticoid therapy, can alleviate the function and coagulation function in patients with endothelial damage, relieve endothelial dysfunction, improve coagulation status of patients, and has higher application value.

Objective To observe the effects of Danqi Granules combined with Xianling Gubao Capsules on bone metabolism and bone mineral density (BMD) in patients with postmenopausal osteoporotic vertebral compression fractures (OVCF) after surgery. Methods 90 cases of postmenopausal women with OVCF were randomly divided into the control group and the observation group, 45 cases in each group. The two groups were treated by vertebroplasty (PVP), and after the surgery, the control group was treated with Xianling Gubao Capsules while the observation group was additionally given Danqi Granules. The serum markers of bone metabolism [(25 (OH) D, PINP, β-CTX] and BMD of L2-4 and left femoral neck were detected before and after treatment. Results The level of serum 25 (OH) D in the observation group after treatment was significantly higher while the levels of PINP and β-CTX were significantly lower than those in the control group (P<0.05), and the BMD of L2-4 and left femoral neck of the observation group were significantly higher than those of the control group (P<0.05). There was no significant difference in the incidence of adverse reactions between the two groups. Conclusion Danqi Granules combined with Xianling Gubao Capsules can significantly accelerate fracture recovery, and improve bone mineral density in the treatment of postmenopausal women with OVCF.

This paper described the measures taken by Sir Run Run Shaw Hospital of Zhejiang University School of Medicine in development of general medicine,and differential care of acute and chronic diseases. Rational allocation of medical resources and leveraging of tertiary hospital′s advantages in medicine and education effectively improved the utilization ratio of medical resources for better health of the people.

Objective To evaluate the feasibility,safety and efficacy of laparoscopic appendectomy for acute appendicitis with previous abdominal surgery.Methods The clinical data of 253 patients with acute appendicitis undergoing laparoscopic surgery was retrospectively studied from Feb.2009 to Jun.2012,including 177 patients without previous abdominal surgery (no previous abdominal surgery group,NPAS group),76 patients with previous abdominal surgery (previous abdominal surgery group,PAS group).Parameters studied were conversion rates,operation time,blood loss,complications rate,length of hospital stay and the intestine function recovery time between two groups.Results The conversion rates were no significant difference between NPAS group and PAS group.The operation time of NPAS group and PAS group was (40.5 ± 12.3) minutes and (62.6 ± 14.2) minutes (P ＜0.05).The blood loss,intestine function recovery time,complications rate,and length of hospital stay were no significant minutes between NPAS group and PAS group after operation (P ＞ 0.05).Conclusions Previous abdominal surgery prolongs the operation time of laparoscopic appendectomy,but history of abdominal surgery has no significant effect on laparoscopic surgical outcome,which may indicate that laparoscopic surgery for acute appendicitis with previous abdominal surgery is safe and effective and still has the adventages of less trauma,faster recovery.

Xanthan gum is a microbial, natural high molecular weight polysaccharide produced by a the bacterium Xanthomonas campestris. Due to its exceptional rheological properties, its numerous areas of application cover a broad range. This review focuses on various aspects of xanthan production, properties, degradation, and application.

Objective To study the impact of maternal high-fat diet during pregnancy and lactation on hepatic steatosis in the early life of offspring rats and its possible mechanism. Methods Female Sprague-Dawley rats were fed either a high fat diet (HF) or control (C) diet for 8 weeks before mating and throughout gestation and ifrst 3 weeks of lactation. The expressions of hepatic fatty acid catabolism related genes, including peroxisome proliferator-activated receptor alpha (PPARα), acyl-CoA syn-thease long-chain family member3 (ACSL3), carnitine palmitoyltransferase-1α(CPT-1α) and 3-hydroxyacyl CoA dehydrogenase (Ehhadh) were determined in offspring liver tissue. The liver pathology was examined in offspring rats at 3 weeks of age. Results Pathohistological ifndings at 3 weeks of age showed that there were diffuse vacuolar degeneration in cytoplasm of hepatocytes and spot necrosis in hepatic lobular in the HF offspring liver. The mRNA expressions of PPARαand Ehhadh genes were markedly increased in the HF offspring as compared to the control group (P<0.05). The mRNA expression of CPT-1αgene was also higher in the HF offspring than that in control group (P=0.19). The level of ACSL3 gene expression, however, was markedly decreased (P<0.05). Conclusions Maternal high fat diet during pregnancy and lactation could result in an increased expression of genes related to hepatic fatty acidβ-oxidation, including PPARα, CPT1αand Ehhadh, but the liver steatosis cannot be reversed in the early life of offspring.

Objective To screen the clinical factors affecting the malignant probability of solitary pulmonary nodule (SPN) with univariate and multivariate Logistic regression analysis,and to establish a clinical prediction model,evaluate its test effectiveness in the differential diagnosis in SPN.Methods A retrospective cohort study included 182 patients with diagnosis of SPN (group A).Clinical data included gender,age,smoking history,quitting smoking,history of tumor,serum carcinoembryonic antigen (CEA),location,diameter,density,vacuole sign,cavity,airbronchogram,calcifcation,lobulation,spiculate sign,spiculation,pleural indentation sign,vascular convergence sign,enhanced CT value,the maximum standard uptake value (SUV~x) of positron-emission tomography (PET),pathological diagnosis were collected.The independent predictors of malignancy were estimated with univariate and multivariate analysis,then the clinical prediction model to identify malignant was established.Other 45 SPN patients (group B) were used to tested value of the model.Results Univariate analysis showed that gender,age,no lower lobe location,diameter,density,airbronchogram,calcification,lobulation,spiculation,pleural indentation sign,enhanced CT value ≥ 15 HU,SUVm,≥ 2.5 of PET significantly affected judgment of SPN of benign or maligant (P ＜ 0.05).Multivariate analysis revealed that female,age,lobulation,short spiculation,pleural indentation sign was the independent predictors of malignancy in patient with SPN,solid nodule suggested benign.The clinical prediction model to identify malignant was established:P =ex/ (1 + ex),x =-3.399 +1.382 × gender + 0.056 × age + 1.377 × lobulation + 1.498 × spiculation-2.096 × solid + 1.005 × pleural indentation sign,e was natural logarithm.P =0.663 was as cut-off point,group B was used to test value of the model:the sensitivity was 86.5％,the specificity was 62.5％,the positive predictive value was 91.4％,the negative predictive value was 50.0％,the accuracy was 82.2％.Conclusions As for SPN,female,age,lobulation,short spiculation and pleural indentation sign is the independent predictor of malignancy in patient with SPN,solid nodule suggests benign.The prediction model is sufficient to estimate the malignancy of patient with SPN.

OBJECTIVETo investigate the effects and mechanisms of Yishen Huoxue decoction (YHD) on chronic renal failure (CRF) rats induced by 5/6 nephrectomy.

METHODSThe glomerulosclerosis model was established by 5/6 nephrectomy in rats. Experimental animals were allocated into the normal group, the model group, the YHD group and the benazepril group. Urine protein of 24 h (UP) at the 6th and 12th weekend after operation, blood urea nitrogen (BUN) and creatinine (SCr), albumin (Alb) and haemoglobin (HB) at the 12th weekend were measured, renal pathology changes were examined with light microscope, the expressions of proliferating cell nuclear antigen (PCNA) and fibronectin (Fn) were examined by immunohistochemistry and mRNA expressions of connective tissue growth factor (CTGF) and plasminogen activator inhibitor-1 (PAI-1) by RT-PCR at the 12th weekend.

RESULTSCompared with those in the normal group, the levels of UP, BUN and SCr, the area of glomerular mesangial matrix, the FN deposition, PCNA expression in glomeruli and tubular interstitium and mRNA expressions of CTGF and PAI-1 were all significantly higher in the model group (P < 0.05). All the above-mentioned indexes were lower in the YHD group than those in the model group (P < 0.05). PCNA positively expressed cells in glomeruli of the normal, model group, YHD group and benazepril group was 7.00 +/- 2.24,34.78 +/- 6.96,15.75 +/- 2.61 and 15.50 +/- 2.57 respectively, positively correlated to the expression of CTGF, PAI-1, FN and SCr level.

CONCLUSIONYHD could delay the progression of CRF in 5/6 nephrectomized rats, and the mechanisms were mainly related to the inhibition on renal cell proliferation and it induced over-expression of cytokines, and accumulation of extracellular matrix.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Extracellular Matrix ; drug effects ; metabolism ; Fibronectins ; biosynthesis ; genetics ; Glomerulosclerosis, Focal Segmental ; drug therapy ; etiology ; metabolism ; Kidney Failure, Chronic ; drug therapy ; etiology ; metabolism ; Male ; Nephrectomy ; Phytotherapy ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

Objective:To establish a method of multi-PCR to amplify the complete DNA sequence (CDS) of TCR ? and ? chain of the antigen-specific T lymphocytes in local pathologic specimen of active pulmonary tuberculosis patients, and to analyze ?/? T cell receptor gene rearrangement and CDR3 repertoire.Methods:The lymphocytes in bronchoalveolar lavage (BAL) of active pulmonary tuberculosis patients were separated. Following total RNA extraction, cDNA synthesis, Multi-PCR, recombinant clones construction, and sequencing, the CDS of TCR ? and ? chains from these lymphocytes were analyzed by using software of DNAstar and internet TCR resources.Results:24 of ? chain CDS and 13 of ? chain CDS from 3 samples of BAL were obtained. As for TCR ? chain, AV1S2 (54%), AV12S3 (41%), and AV12S2(5%) appeared frequently. BV2(38%), BV29S1(46%), BV14(3%), and BV4S2(3%) in TCR ? chain appeared more often. There were CDR3 diversities between samples and even in the same sample by amino acid sequence analysis, but there were a few identical or similar amino acid sequences. There was the same amino acid sequence of SVGTGTLHQETQY in CDR3 region of ? chain of BAL sample No.1 and No.2; The sequence of AVRDWAGNMLT appeared in two ? chains of BAL sample No.2 and No.3; Moreover, the sequence of AV…DNN…RLM appeared in ? chains of BAL sample No.2 and No.3.Conclusion:A method of Multi-PCR is used to amplify TCR ? and ? chain CDS of tuberculosis patients. There are characteristic T cell clones to proliferate,with TCR ? and ? chain repertiore skewing in local infective focus. The sequences of CDR3 in different TCR clones are mostly different but there are a few identical or similar sequences in the same patient or even between different patients. The identical amino acid sequences of CDR3 are possibly specific for recognizing MTB polypeptide.