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 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 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 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.

Objective To compare the therapeutic effect of video-assisted thoracoscopic surgery and traditional thoracotomy in fixation of traumatic multiple rib fractures.Methods Clinical data of 56 patients with traumatic multiple rib fractures treated surgically between July 2005 and September 2012 were analyzed retrospectively.Based on the treatments,the patients were assigned to video-assisted thoracoscopy group (thoracoscopy group,n =27) and traditional thoracotomy group (thoracotomy group,n =29).A comparison was done on the variables including operation time,intraoperative blood loss,ventilator support rate,duration of mechanical ventilation,length of ICU stay,incidence of lung infections,visual analogue scale (VAS) at day 3 postinjury and mortality between the two groups.Results Operation time [(128.9 ± 21.1) min vs (140.7 ± 24.2) min],ventilator support rate (70％ vs 76％) and mortality (4％ vs 7％) in thoracoscopy group revealed no statistical differences compared with thoracotomy group (P ＞ 0.05),but intraoperative blood loss [(321.1 ± 30.1)ml vs (438.1 ± 43.2)ml],duration of mechanical ventilation [(4.3 ± 2.1) d vs (7.2 ± 1.6) d],length of ICU stay [(5.9 ± 21.1) d vs (8.5 ± 1.7) d],incidence of lung infection (33％ vs 90％),and VAS [(7.0 ± 1.4) points vs (8.3 ± 0.9) points] were significantly reduced in thoracoscopy group than in thoracotomy group (all P ＜ 0.01).Conclusion Video-assisted thoracoscopic surgery is characterized by fewer intraoperative bleeding,shorter duration of mechanical ventilation and ICU stay,and lower lung infection rate during treatment of traumatic multiple rib fractures compared to traditional thoracotomy.

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 The action mechanisms of Qingfei Oral Liquid ( QFOL) in the treatment of respiratory syncytial virus ( RSV) infection need to be studied more deeply.The aim of this study is to examine the expressions of interleukin ( IL)-10 and IL-17 in the lung tissue and those of Treg and Th17 in the spleen tissue of RSV-infected mice before and after treated with QFOL, and to explore the action mechanisms of QFOL from the perspective of the Treg/Th17 cy-tokines balance. Methods Fifty BABL/c mice were equally ran-domized to five groups: blank control, RSV model, Ribavirin, low-dose QFOL, and high-dose QFOL.Models of RSV ( long strain) infec-tion were made in the latter four groups.At 48 hours after viral activa-tion, the mice of the control and RSV model groups were treated intragastrically with 0.9%normal saline and those in the Ribavirin and QFOL groups with Ribavirin at 0.0025 g/mL and QFOL at 1.33 g/mL and 4 g/mL, respectively, all for 72 hours.Then all the mice were killed and the lung tissue harvested from 5 animals in each group for pathological analysis, while the levels of IL-10 and IL-17 in the bronchoalveolar lavage fluid of the other 5 detected by ELISA.The expressions of the cytokines Treg and Th17′in the spleen from 4 mice in each group were determined by flow cytometry. Results Compared with the RSV models, pathologic changes were significantly re-duced in the mice of the QFOL, Ribavirin and control groups (P<0.01), the expression of IL-10 remarkably up-regulated in the low-dose QFOL, high-dose QFOL, Ribavirin, and control groups ([39.21 ±1.57] vs [43.54 ±1.03], [46.64 ±0.48], [47.83 ±0.87], and [50.44 ±1.04] ng/L, all P<0.01), while the level of IL-17 markedly down-regulated ([70.96 ±0.53] vs [55.92 ±0.83], [33.66 ±0.70], [21.92 ±1.38], and [9.42 ±0.59] pg/mL, all P<0.01).The expressions of Treg/Th17′were significantly in-creased in the low-dose QFOL, high-dose QFOL, Ribavirin, and control groups (2.89 ±0.52, 6.38 ±0.36, 3.95 ±0.26, and 3.54 ± 0.85) as compared with that in the RSV models (0.96 ±0.16) (all P<0.01).Both low-and high-dose QFOL groups showed statisti-cally significant differences from the Ribavirin group in the levels of Treg, Th17, and Treg/Th17 (P<0.05). Conclusion QFOL can regulate the balance of Treg/Th17, increase the expression of IL-10 and decrease that of IL-17 in the lung tissue of RSV-infected mice, which further proves the efficacy of QFOL in the treatment of RSV-induced pneumonia.

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 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.