Objective To evaluate the effect of remifentanil on Toll-like receptor 2 (TLR2) mRNA expression during renal ischemia-reperfusion (Ⅰ/R) in rats.Methods Fifty-four male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 3 groups (n=18 each) using a random number table:sham operation group (group S),Ⅰ/R group and remifentanil group (group R).Renal Ⅰ/R injury was produced by clamping the bilateral renal arteries for 45 min followed by reperfusion in Ⅰ/R and R groups.Bilateral renal arteries were only exposed but not clamped in group S.Remifentanil 1.0 μg · kg-1 · min-1 was infused via the tail vein starting from 15 min before ischemia until 30 min of reperfusion in group R,while the equal volume of normal saline was given instead in S and Ⅰ/R groups.The animals were sacrificed at 15 min before ischemia and 6 and 24 h of reperfusion,and the renal specimens were obtained for examination of the pathological changes (with light microscope) and for determination of the contents of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) (by ELISA) and expression of TLR2 mRNA (by RT-PCR) and cell apoptosis (by double staining and flow eytometry).The apoptotic rate was calculated.Results Compared with group S,TLR2 mRNA expression was significantly up-regulated,and the contents of TNF-α and IL-6 and apoptotic rate were increased at 6 and 24 h of reperfusion in Ⅰ/R and R groups.Compared with group Ⅰ/R,TLR2 mRNA expression was significantly down-regulated,and the contents of TNF-α and IL-6 and apoptotic rate were decreased at 6 and 24 h of reperfusion in group R.The pathological changes were significantly attenuated in group R as compared with group Ⅰ/R.Conclusion The mechanism by which remifentanil reduces renal Ⅰ/R injury is related to down-regulation of TLR2 expression and decrease in TLR2 activity and inhibition of inflammatory responses in renal tissues and cell apoptosis in rats.

Objective To investigate risk factors related to postoperative death of patients with rectosigmoid junction tumor perforation.Methods The clinical data of 76 cases with rectosigmoid junction tumor perforation confirmed by laparotomy from January 2000 to October 2015 were collected.Results Of the 76 cases,17 patients died postoperatively,the mortality rate was 22％,the single factor analysis showed that age(x2 =4.649,P =0.031),duration of abdominal pain(x2 =8.218,P =0.016),severe heart and lung diseases(x2 =11.996,P =0.007),circulatory and renal function(x2 =10.360,P =0.016),serum albumin(x2 =7.252,P =0.027),white blood cell count(x2 =7.633,P =0.022),Perforation diameter (x2 =9.770,P =0.008),Geroge grade of intraperitoneal contamination (x2 =10.086,P =0.006) were related to postoperative death (P ＜ 0.05).Multivariate analysis showed that complicating severe heart and lung diseases,preexisted circulatory and renal dysfunction,white blood cell count ＜ 4 × 109/L,size ＞ 3 cm,intraperitoneal contamination larger than one quadrant were independent risk factors for postoperative death.Conclusion Risk factors related to postoperative death of rectosigmoid junction tumor perforation were preoperative important organ dysfunction and intraperitoneal infection.

Objective To investigate the efficacies of different times and frequencies of acupoint application in preventing and treating child asthma of spleen-lung qi deficiency type. Method Sixty children with asthma of spleen-lung qi deficiency type were randomly allocated to dog days, dog days reinforcement, coldest days and coldest days reinforcement groups for treatment by acupoint application. The therapeutic effects were evaluated using the TCM symptom score as the observation indicator in the four groups after three months of treatment. Result The TCM symptom score decreased significantly in the four groups at the follow-up after three months of treatment (P<0.05). The main effect of treatment time was marked (F=17.04, P<0.05). The main effect of treatment frequency was marked (F=8.17, P<0.05). Conclusion Different times and frequencies of acupoint application both decrease the TCM symptom score in children with asthma. The therapeutic effect is better in dog days than in three nine-day period after the winter solstice. An increase in treatment frequency influences the therapeutic effect obviously. The therapeutic effect is best in the dog days reinforcement group.

Objective:To explore the inhibition of microRNA (miRNA, miR)-5590-3p on the expression of transforming growth factor beta typeⅡreceptor (TGFBR2) gene and its effect on the invasion and proliferation of gastric cancer cell line HS-746T.Methods:The gastric cancer cell line was cultured in vitro. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to analyze the expression of miR-5590-3p in gastric cancer tissues and cell lines. The miR-NC and miR-5590-3p mimic were transfected into gastric cancer cell line HS-746T by lipofection, and named as miR-NC group and miR-5590-3p group, respectively. qRT-PCR was used to measure transfection effects. Transwell assay and cell counting kit-8 (CCK-8) assay were used to detect cell invasion and proliferation after transfection. The bioinformatics software predicts and the dual luciferase reporter gene system validates the target gene of miR-5590-3p. qRT-PCR and western blot were used to measure the expression of TGFBR2 and its downstream proteins in the transfected cells. Results:The expression of miR-5590-3p in gastric cancer tissues was significantly lower than that in adjacent tissues ( P<0.01). The expression of miR-5590-3p in gastric cancer cell lines was significantly lower than that in normal gastric mucosal epithelial cells ( P<0.05), and lowest in HS-746T cells ( P<0.01). After transfection, the expression of miR-5590-3p in miR-5590-3p group (11.76±0.21) was significantly higher than that in miR-NC group (1.06±0.21), with statistically significant difference ( P<0.01). The number of invasive cells in miR-NC group and miR-5590-3p group were (101.20±15.47) and (26.53±6.53), respectively, and the invasion ability of miR-5590-3p group was significantly decreased ( P<0.01). Compared with the miR-NC group, the cell proliferation ability of the miR-5590-3p group was significantly decreased ( P<0.05). Bioinformatics software showed that the target gene for miR-5590-3p is TGFBR2. The dual luciferase reporter gene system confirmed that miR-5590-3p can target the TGFBR2 gene ( P<0.01). Western blot results showed that compared with miR-NC group, the expression of TGFBR2 in HS-746T cells in miR-5590-3p group was significantly decreased ( P<0.01), the expression of ZEB-1 and ZEB-2, and the expression of CDK1 and cyclin B proteins were decreased as well. Conclusions:miR-5590-3p can inhibit the invasion and proliferation of gastric cancer cell HS-746T by targeting and regulating the expression of TGFBR2 gene.

AIM To study the effects of Rehmanniae Radix Preparata (A) and Poria (B) on decoction amounts of loganin,morroniside and 5-hydroxymethylfurfural in Corni Fructus (C).METHODS These three medicinal materials were combined one another and divided into seven groups (A,B,C,A + B,A + C,B + C and A + B + C).Then the contents of three constituents were determined by HPLC.RESULTS Compared with the single decoction of Corni Fructus,the decoction amounts of loganin and 5-hydroxymethylfurfural were decreased,and that of morroniside was increased at the time of mixed decoction of Rehmanniae Radix Preparata and Corni Fructus,or Rehmanniae Radix Preparata,Poria and Corni Fructus.This situation was just the contrary at the time of mixed decoction of Poria and Corni Fructus.CONCLUSION The mixed decoction of Corni Fructus,Rehmanniae Radix Preparata and Poria increases the decoction amount of morroniside,which may make mixed decoction liquid show better efficacy.

OBJECTIVE:To study the effects of combined decoctions of alismatis rhizoma and curcumae radix and rehmanniae radix praeparata on the decoction amount of acteoside in rehmanniae radix praeparata,and provide reference for studying the effec-tive ingredients of three-drug effect. METHODS:Single decoction-1(rehmanniae radix praeparata 20 g),single decoction-2(alis-matis rhizoma 20 g),single decoction-3(curcumae radix 20 g),combined decoction-1(rehmanniae radix praeparata 20 g,alisma-tis rhizoma 20 g),combined decoction-2(rehmanniae radix praeparata 20 g,curcumae radix 20 g),combined decoction-3(alisma-tis rhizoma 20 g,curcumae radix 20 g)and combined decoction-4(rehmanniae radix praeparata 20 g,alismatis rhizoma 20 g,cur-cumae radix 20 g)were respectively taken to prepare dry extracts after extracting by refluxing,HPLC was used to detect the acteo-side content and calculate its decoction amount in sample. RESULTS:The decoction amounts of acteoside in single decoction-1, combined decoction-1,combined decoction-2 and combined decoction-4 dry extracts were 0.0354,0.0223,0.0228,0.0110 mg/g,respectively. Compared with single decoction-1 group,the last 3 groups had statistical significances(P<0.01);combined decoc-tion-4 showed lowest decoction amount in three-drug combined decoction group. Acteoside was not detected in the negative control groups(single decoction-2,single decoction-3 and combined decoction-3). CONCLUSIONS:The decoction amount of acteoside is reduced when alismatis rhizoma,curcumae radix and rehmanniae radix praeparata were decocted together,indicating that it may not be the main ingredient of playing effects in three-drug combined decoction liquid.

Objective Dendritic cell-associatedC-type lectin-1 ( Dectin-1) is one of the most important receptors in antifungal innate immune response.This study was to construct a recombinant adenovirus vector expressing themurine Dectin-1gene and acquire a high-concentration adenovirus by amplification and purification. Methods The PCR amplification product CLEC7A-pIRES2-EGFP was cloned into the intermediate vector pDONR221, and then recom-bined with the backbone vector pAD/CMV/V5-DEST to produce a re-combinant plasmid pAD-CLEC7A-pIRE2S -EGFP.The recombinant plasmid was linearized with Pac I and transfected into human embryon-ic kidney ( HEK293) cells to produce recombinant adenovirus pAD-CLEC7Ap-IRES 2-EGFP. The adenovirus was propagated in the HEK293 cells and purified by filtering through the cellulose acetate membrane and concentrating column.Fluorescence microscopy and re-al-time PCR were used to determine the expression of the Dectin-1 gene. Results PCR identification, enzyme digestion, and sequen-cing results manifested theDectin-1 gene in the vector, with the final adenovirus titer of 5×1011 IU/mL.Fluorescence microscopy revealed green fluorescence and real-time PCR assay confirmed that the expression of Dectin-1 was improved by 8677.25 times. Conclusion A relatively high-titer adenovirus expressing Dectin-1 was acquired,which may help to further study the high expression of Dectin-1 in anti-fungal innate immunity in vitro and in vivo.

Objective To investigate of the risk factors for the death of severe fever with thrombocytopenia syndrome (SFTS),so as to set up SFTS critical score and evaluate its role in predicting the prognosis for patients with SFTS.Methods A total of 123 SFTS patients hospitalized in Ji′nan Hospital of Infectious Diseases affiliated to Shandong University from June 2011 to October 2014 were enrolled in this study.The univariate Logistic regression analysis was performed to analysis the risk factor for the death of SFTS.Then the SFTS critical score system was set up accordingly.The prognosis value of SFTS critical score was compared with the rapid emergency medicine score (REMS)and the acute physiology and chronic health evaluation Ⅱ (APACHEⅡ)by using receiver operator characteristic curve (ROC).Results Among all the patients,17 males and 14 females were in death group,and 45 males and 47 females were in survival group.The results of the univariate Logistic regression analyses indicated that the glasgow coma scale (GCS),lactate dehydrogenase,activated partial thromboplastin time,oxygen saturation were risk factors for the death of SFTS,with statistically significant difference (all P <0.05). All of the four parameters of SFTS critical scores in the death group were higher than those in the survival group,with statistically significant difference (all P <0.05 ).The REMS,APACHEⅡ score and SFTS critical score in the death group were significantly higher than those in the survival group (all P <0.01 ). The area under the curve (AUC)of REMS,APACHE Ⅱ scores and SFTS critical score were 0.734, 0.746 and 0.788,respectively.The Youden index of the SFTS critical scores was the highest among all three scores (P <0.01).If 15 .0 was used as the cut off value of SFTS critical score,the specificity and the sensitivity for predicting the death risk for the hospitalized patient were 74.2% and 76.1 %, respectively.Conclusion SFTS critical score,REMS and APACHEⅡ score can all effectively predict the prognosis for SFTS patients,among which,the SFTS critical score is the most convenient and has the best predictive value.

OBJECTIVE:To ensure the stability of electronic data capture(EDC)system in drug clinical trials and to improve the quality of drug clinical trials. METHODS:The quality control system for EDC system was established and introduced from the formulation of quality control process,establishment of data standard,trial project management,daily management,trial project design,system operation,system function,etc. RESULTS & CONCLUSIONS:Data standard have been achieved through estab-lishing EDC quality control system by our hospital based on attributable,legible,contemporaneous,original and accurate principle. The management of trial project and daily management are conducted through data registration,staff training,the formulation of da-ta management plan,fault emergency treatment,database backup;multiple verification of support data,data lock and export,trial report autogeneration and other functions have been realized by formulating related standard operation instruction,program file,op-eration manual and quality record. Those aspects improve facticity,accuracy and integrality of data in clinical trials,and lay a foun-dation for further data mining.

Objective:To study the effects and mechanisms of acetyl-11-keto-β-boswellic acid ( AKBA) in myocardial ischemic model induced by isoproterenol hydrochloride ( ISO) in rats. Methods:The SD rats were randomly divided into the sham group, model group, AKBA low dose group and AKBA high dose group. Myocardial injury model was induced by subcutaneous injection of ISO (100 mg·kg-1 ) . The change of ST segment in ECG was observed. Creatine kinase ( CK-MB) , cardiac troponin I ( cTnI) , lactate dehydro-genase( LDH) , malondialdehyde( MDA) and superoxide dismutase( SOD) in the blood were detected by ELISA. The change of histo-logical tissue was determined by HE staining, and cell apoptosis was analyzed by TUNEL assay. Results: Serum CK-MB, cTnI and LDH were decreased significantly in AKBA high dose group when compared with those in the model group. Compared with that in the model group, MDA content was lowered and the SOD activity was increased in AKBA high dose group. Furthermore, AKBA high dose group improved the pathologic changes of myocardium. TUNEL assay revealed significant reduction of cardiomyocytes apoptosis in the hearts of the ischemic rats in AKBA high dose group. Conclusion:AKBA has excellent cardioprotective effect on myocardial ischemic induced by ISO and protection of myocardial cells from injury.