AIM:To establish a mouse model of chronic obstructive pulmonary disease(COPD)induced by cigarette smoke(CS)exposure combined with polyinosinic-polycytidylic acid(Poly I:C)nasal drip,and to investigate the mechanism of airway epithelial barrier injury in COPD.METHODS:(1)Ninety-six male BALB/c mice were randomly divided into control group,CS group,Poly I:C group,and CS+Poly I:C group(n=24).The model was established from week 1 to week 8,with pulmonary function tested every 4 weeks.Six mice from each group were sacrificed at the end of weeks 4,8,16,and 24.Changes in minute volume(MV),enhanced pause(Penh),mean linear intercept(MLI)and bronchial wall thickness(BWT)were observed.The protein levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),zonula occludens-1(ZO-1)and E-cadherin(E-Cad)in the lung were detected.(2)Human bronchial epithe-lial BEAS-2B cells were stimulated with CS extract(CSE)combined with Poly I:C for 24 h,and then the protein levels of occludin(Occ),ZO-1,and phosphorylated epidermal growth factor receptor(EGFR),P38 and extracellular signal-regu-lated kinase(ERK)1/2 were analyzed.RESULTS:(1)Compared with control group,at the 8th week,the mice in CS and CS+Poly I:C groups showed typical pathological changes in lung tissues,including significant inflammatory cell infil-tration,alveolar cavity expansion,alveolar wall rupture and fusion,and airway wall thickening.The Penh,BWT,MLI,and lung IL-1β and TNF-α levels were significantly increased(P<0.05 or P<0.01),while MV and lung ZO-1 and E-Cad levels were remarkably decreased(P<0.05 or P<0.01).By the 24th week,these pathological changes remained relative-ly stable in CS+Poly I:C group.(2)Compared with control group,CSE and its combination with Poly I:C dramatically in-duced a reduction in ZO-1 and Occ protein expression in BEAS-2B cells(P<0.05 or P<0.01),and increased the levels of phosphorylated EGFR,P38 and ERK1/2(P<0.01).The effects in CSE combined with Poly I:C group were considerably superior to those in CSE or Poly I:C group alone.CONCLUSION:Poly I:C can enhance the pathological changes and airway epithelial barrier damage induced by CS in a mouse model of COPD,which may be related to the activation of EGFR/ERK/P38 signaling pathway.

Objective:To investigate the epidemiological characteristics, diagnosis, treat-ment and prognosis of gallbladder cancer in China from 2010 to 2017.Methods:The single disease retrospective registration cohort study was conducted. Based on the concept of the real world study, the clinicopathological data, from multicenter retrospective clinical data database of gallbladder cancer of Chinese Research Group of Gallbladder Cancer (CRGGC), of 6 159 patients with gallbladder cancer who were admitted to 42 hospitals from January 2010 to December 2017 were collected. Observation indicators: (1) case resources; (2) age and sex distribution; (3) diagnosis; (4) surgical treatment and prognosis; (5) multimodality therapy and prognosis. The follow-up data of the 42 hospitals were collected and analyzed by the CRGGC. The main outcome indicator was the overall survival time from date of operation for surgical patients or date of diagnosis for non-surgical patients to the end of outcome event or the last follow-up. Measurement data with normal distribu-tion were represented as Mean±SD, and comparison between groups was conducted using the t test. Measurement data with skewed distribution were represented as M( Q1, Q3) or M(range), and com-parison between groups was conducted using the U test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Univariate analysis was performed using the Logistic forced regression model, and variables with P<0.1 in the univariate analysis were included for multivariate analysis. Multivariate analysis was performed using the Logistic stepwise regression model. The life table method was used to calculate survival rates and the Kaplan-Meier method was used to draw survival curves. Log-rank test was used for survival analysis. Results:(1) Case resources: of the 42 hospitals, there were 35 class A of tertiary hospitals and 7 class B of tertiary hospitals, 16 hospitals with high admission of gallbladder cancer and 26 hospitals with low admission of gallbladder cancer, respectively. Geographical distribution of the 42 hospitals: there were 9 hospitals in central China, 5 hospitals in northeast China, 22 hospitals in eastern China and 6 hospitals in western China. Geographical distribution of the 6 159 patients: there were 2 154 cases(34.973%) from central China, 705 cases(11.447%) from northeast China, 1 969 cases(31.969%) from eastern China and 1 331 cases(21.611%) from western China. The total average number of cases undergoing diagnosis and treatment in hospitals of the 6 159 patients was 18.3±4.5 per year, in which the average number of cases undergoing diagnosis and treatment in hospitals of 4 974 patients(80.760%) from hospitals with high admission of gallbladder cancer was 38.8±8.9 per year and the average number of cases undergoing diagnosis and treatment in hospitals of 1 185 patients(19.240%) from hospitals with low admission of gallbladder cancer was 5.7±1.9 per year. (2) Age and sex distribution: the age of 6 159 patients diagnosed as gallbladder cancer was 64(56,71) years, in which the age of 2 247 male patients(36.483%) diagnosed as gallbladder cancer was 64(58,71)years and the age of 3 912 female patients(63.517%) diagnosed as gallbladder cancer was 63(55,71)years. The sex ratio of female to male was 1.74:1. Of 6 159 patients, 3 886 cases(63.095%) were diagnosed as gallbladder cancer at 56 to 75 years old. There was a significant difference on age at diagnosis between male and female patients ( Z=-3.99, P<0.001). (3) Diagnosis: of 6 159 patients, 2 503 cases(40.640%) were initially diagnosed as gallbladder cancer and 3 656 cases(59.360%) were initially diagnosed as non-gallbladder cancer. There were 2 110 patients(34.259%) not undergoing surgical treatment, of which 200 cases(9.479%) were initially diagnosed as gallbladder cancer and 1 910 cases(90.521%) were initially diagnosed as non-gallbladder cancer. There were 4 049 patients(65.741%) undergoing surgical treatment, of which 2 303 cases(56.878%) were initially diagnosed as gallbladder cancer and 1 746 cases(43.122%) were initial diagnosed as non-gallbladder cancer. Of the 1 746 patients who were initially diagnosed as non-gallbladder cancer, there were 774 cases(19.116%) diagnosed as gallbladder cancer during operation and 972 cases(24.006%) diagnosed as gallbladder cancer after operation. Of 6 159 patients, there were 2 521 cases(40.932%), 2 335 cases(37.912%) and 1 114 cases(18.087%) undergoing ultrasound, computed tomography (CT) or magnetic resonance imaging (MRI) examination before initial diagnosis, respec-tively, and there were 3 259 cases(52.914%), 3 172 cases(51.502%) and 4 016 cases(65.205%) undergoing serum carcinoembryonic antigen, CA19-9 or CA125 examination before initially diagnosis, respectively. One patient may underwent multiple examinations. Results of univariate analysis showed that geographical distribution of hospitals (eastern China or western China), age ≥72 years, gallbladder cancer annual admission of hospitals, whether undergoing ultrasound, CT, MRI, serum carcinoembryonic antigen, CA19-9 or CA125 examination before initially diagnosis were related factors influencing initial diagnosis of gallbladder cancer patients ( odds ratio=1.45, 1.98, 0.69, 0.68, 2.43, 0.41, 1.63, 0.41, 0.39, 0.42, 95% confidence interval as 1.21-1.74, 1.64-2.40, 0.59-0.80, 0.60-0.78, 2.19-2.70, 0.37-0.45, 1.43-1.86, 0.37-0.45, 0.35-0.43, 0.38-0.47, P<0.05). Results of multivariate analysis showed that geographical distribution of hospitals (eastern China or western China), sex, age ≥72 years, gallbladder cancer annual admission of hospitals and cases undergoing ultrasound, CT, serum CA19-9 examination before initially diagnosis were indepen-dent influencing factors influencing initial diagnosis of gallbladder cancer patients ( odds ratio=1.36, 1.42, 0.89, 0.67, 1.85, 1.56, 1.57, 0.39, 95% confidence interval as 1.13-1.64, 1.16-1.73, 0.79-0.99, 0.57-0.78, 1.60-2.14, 1.38-1.77, 1.38-1.79, 0.35-0.43, P<0.05). (4) Surgical treatment and prognosis. Of the 4 049 patients undergoing surgical treatment, there were 2 447 cases(60.435%) with complete pathological staging data and follow-up data. Cases with pathological staging as stage 0, stage Ⅰ, stage Ⅱ, stage Ⅲa, stage Ⅲb, stage Ⅳa and stage Ⅳb were 85(3.474%), 201(8.214%), 71(2.902%), 890(36.371%), 382(15.611%), 33(1.348%) and 785(32.080%), respectively. The median follow-up time and median postoperative overall survival time of the 2 447 cases were 55.75 months (95% confidence interval as 52.78-58.35) and 23.46 months (95% confidence interval as 21.23-25.71), respectively. There was a significant difference in the overall survival between cases with pathological staging as stage 0, stage Ⅰ, stage Ⅱ, stage Ⅲa, stage Ⅲb, stage Ⅳa and stage Ⅳb ( χ2=512.47, P<0.001). Of the 4 049 patients undergoing surgical treatment, there were 2 988 cases(73.796%) with resectable tumor, 177 cases(4.371%) with unresectable tumor and 884 cases(21.833%) with tumor unassessable for resectabi-lity. Of the 2 988 cases with resectable tumor, there were 2 036 cases(68.139%) undergoing radical resection, 504 cases(16.867%) undergoing non-radical resection and 448 cases(14.994%) with operation unassessable for curative effect. Of the 2 447 cases with complete pathological staging data and follow-up data who underwent surgical treatment, there were 53 cases(2.166%) with unresectable tumor, 300 cases(12.260%) with resectable tumor and receiving non-radical resection, 1 441 cases(58.888%) with resectable tumor and receiving radical resection, 653 cases(26.686%) with resectable tumor and receiving operation unassessable for curative effect. There were 733 cases not undergoing surgical treatment with complete pathological staging data and follow-up data. There was a significant difference in the overall survival between cases not undergoing surgical treatment, cases undergoing surgical treatment for unresectable tumor, cases undergoing non-radical resection for resectable tumor and cases undergoing radical resection for resectable tumor ( χ2=121.04, P<0.001). (5) Multimodality therapy and prognosis: of 6 159 patients, there were 541 cases(8.784%) under-going postoperative adjuvant chemotherapy and advanced chemotherapy, 76 cases(1.234%) under-going radiotherapy. There were 1 170 advanced gallbladder cancer (pathological staging ≥stage Ⅲa) patients undergoing radical resection, including 126 cases(10.769%) with post-operative adjuvant chemotherapy and 1 044 cases(89.231%) without postoperative adjuvant chemo-therapy. There was no significant difference in the overall survival between cases with post-operative adjuvant chemotherapy and cases without postoperative adjuvant chemotherapy ( χ2=0.23, P=0.629). There were 658 patients with pathological staging as stage Ⅲa who underwent radical resection, including 66 cases(10.030%) with postoperative adjuvant chemotherapy and 592 cases(89.970%) without postoperative adjuvant chemotherapy. There was no significant difference in the overall survival between cases with postoperative adjuvant chemotherapy and cases without postoperative adjuvant chemotherapy ( χ2=0.05, P=0.817). There were 512 patients with pathological staging ≥stage Ⅲb who underwent radical resection, including 60 cases(11.719%) with postoperative adjuvant chemotherapy and 452 cases(88.281%) without postoperative adjuvant chemotherapy. There was no significant difference in the overall survival between cases with postoperative adjuvant chemo-therapy and cases without post-operative adjuvant chemo-therapy ( χ2=1.50, P=0.220). Conclusions:There are more women than men with gallbladder cancer in China and more than half of patients are diagnosed at the age of 56 to 75 years. Cases undergoing ultrasound, CT, serum CA19-9 examination before initial diagnosis are independent influencing factors influencing initial diagnosis of gallbladder cancer patients. Preoperative resectability evaluation can improve the therapy strategy and patient prognosis. Adjuvant chemotherapy for gallbladder cancer is not standardized and in low proportion in China.

Objective: To explore the best surgical timing after neoadjuvant chemoradiation for advanced rectal cancer patients. Methods: According to the time interval between neoadjuvant chemoradiation and surgery, 117 patients with advanced rectal cancer were divided into short interval group (≤7 weeks, n=54) and long interval group (>7 weeks, n=64). The endpoints included postoperative pathology, short-term efficacy, tumor recurrence and patient survival between the two groups. Results: There were 8 cases PCR in short interval group and 20 cases in long interval group(P=0.415). There were 23 cases of T downgrade in short interval group and 40 cases in the long interval group, which has significant difference (P=0.039). There were 21 cases of N downgrade in short interval group and 38 cases in long interval group, which has significant different (P=0.033). The short-term group was effective in 28 cases, stable in 20 cases, and progressed in 5 cases. In short term efficacy comparison, the cases of complete response, stable disease and progressive disease in short interval group was 28 cases, 20 and 5, long interval group was 47 cases, 14 cases and 3 cases, which has no significant difference(P=0.068). The 3-year local recurrence rate of short interval group and long interval group was 17.0% and 4.7%, respectively, and the difference was statistically significant(P=0.029). The incidence of recurrence in 3 years of short interval group and long interval group was 64.2% and 79.7%, respectively, and the difference was not significant (P=0.061). The highest PCR rate was reached in the 10th and 11th week after neoadjuvant chemoradiotherapy. Of the 12 and 8 patients who underwent surgery, 3 (25.0%) and 2 (25.0%) achieved PCR, respectively. Conclusion PCR and local recurrence rate might be improved by time interval between neoadjuvant chemoradiation and surgery was more than 7 weeks.

Aim To investigate the effects of aspirin on Epstein-Barr virus (EBV)-transformed human B-lym-phocytes.Methods EBV-transformed human B-lym-phocytes were treated with certain concentrations of as-pirin.Cellular proliferation was analyzed by MTT as-say.Further evaluation of apoptosis of aspirin-treated cells was performed through light-field microscope, transmission electronic microscope(TEM),propidium iodide(PI)staining and flow cytometric analysis and DNA electrophoresis. Finally, immunoblot analysis was used to determine the expression levels of apopto-sis-associated proteins, proteins involved in mTOR pathway and PU.1 -Bim axis.Results Aspirin treat-ment inhibited proliferation of EBV-transformed human B-lymphocytes.We observed that aspirin treatment in-duced apoptosis in EBV-transformed human B-lympho-cytes,resulting in the decreased number and size of cells.Ultramicroscopic structural analysis via TEM in-dicated that aspirin treatment deformed the cellular nu-cleus,and led to peripheral chromatin and cytoplasmic vacuole.PI staining and flow cytometric analysis indi-cated that aspirin increased the permeability of cell membrane and decreased the viability of treated cells. Agarose electrophoresis revealed DNA smear in aspirin-treated cells.Mechanistically,mTOR signaling was in-hibited in aspirin-treated cells,as evidenced by the de-creased phosphorylation of S6K1 and S6 via immunob-lot analysis.Aspirin treatment led to the decrease of hematopoietic transcription factor PU.1 .Consequently, pro-apoptotic Bim, apoptosis-associated proteins caspase-3 and PARP were activated in aspirin-treated cells.Conclusion Aspirin may show anti-lymphoma effects via its inhibition of proliferation and induction of apoptosis of EBV-transformed human B-lymphocytes, in which mTOR signal pathway and PU.1 -Bim axis may be involved.

BACKGROUND:A variety of factors contribute to biliary injury that is difficult to be repaired. Stent implantation is extensively used for bile duct injury, but either scaffolds made by metal or plastics can lead to certain adverse reactions. OBJECTIVE:To explore the biological characteristics of a novel biodegradable scaffold and its repair effects on bile duct injury. METHODS:The biological characteristics of the novel biodegradable scaffold were detected by fresh bile, and its degradation was observed at different time points. Thirty Bama mini pigs were included and were randomly divided into observation group (n=15) and control group (n=15). After bile duct injury models were prepared, the control group was subjected to the bile duct interrupted suture, while the observation group was subjected to the novel biodegradable scaffold combined with omentum majus. The biological properties of the scaffolds were observed. Hepatic enzymes and serum total bilirubin levels were detected, as wel as hematoxylin-eosin staining, Masson staining and immunohistochemistry detection ofα-smooth muscle actin were performed. RESULTS AND CONCLUSION:Before and 1, 3 and 6 months after surgery, hepatic enzymes and total bilirubin of two groups were detected, and neither intra-group nor intergroup comparisons had significant differences (P>0.05). Hematoxylin-eosin staining and Masson staining revealed that inflammatory reactions and fiber hyperplasia at the anastomotic site in the observation group were lighter than those in the control group at different time points after surgery. Theα-smooth muscle actin-positive scores in both two groups were in a rise at 1 and 3 months after surgery, and peaked at the 3rd month, and then began to decline. Moreover, theα-smooth muscle actin-positive scores in the observation group were significantly lower than those in the control group at 3 and 6 months after surgery (P<0.05). These results show that the novel biodegradable scaffold has good biological characteristics and can obtain ideal repair effects in the bile duct injury.

Objective To isolate and identify viruses from fecal samples of tree shrew with diarrhea.Methods Fecal sample supernatant of tree shrew with diarrhea was inoculated to three cell lines ( Vero, LLC-MK2 and KMB17 ) , and the cytopathic effects on the cells were observed.The infectious particles in the culture supernatant were further ana-lyzed by transmission electron microscopy ( TEM) , genomic RNA-PAGE, rotavirus detection kit, amplification of S1 com-plete segment and bioinformatics analysis.Results Constant cytopathic effects were induced in Vero, LLC-MK2 and KMB17 cell lines after three passages of culture.The results from TEM, RNA-PAGE and rotavirus analysis indicated that they belong to reoviruses.Analysis of the S1 segments revealed that the S1 sequence from KMB17 cell culture had the high-est homology with that of prototype isolate T1L (85%nucleotide homology and 90%amino acid homology), therefore this isolate was named as type I reovirus.The other two S1 sequences from LLC-MK2 and Vero cell culture were identical to have 85%nucleotide homology and 92%amino acid homology with the prototype isolate T3D, named as type III reovirus. Phylogenetic analysis indicated that the isolates in this study are evolutionally adapted to tree shrews.Conclusions It is the first report here that 2 genotypes of Tupaia orthoreovirus are isolated and identified from one fecal sample via three cell lines and viral S1-specific primers, which provides useful guidelines for the isolation and identification of other reoviruses from tree shrew or other hosts.

Objective To explore the changes of mTOR signaling in LPS/D-gal-induced acute hepatitis in mice . Methods Twenty-six healthy adult female ICR mice were divided into two groups:the control group and experimental group, 13 mice in each group .LPS/D-gal was used to induce acute hepatitis in the mice .The survival of mice was moni-tored within 24 hours after LPS/D-gal challenge .At 6 hours after challenge , samples of serum and liver tissue were collect-ed for further analysis.Results Injection of LPS/D-gal resulted in acute death of the mice within 24 hours.At 6 hours post LPS/D-gal injection , the blood levels of ALT and AST were significantly increased .The mRNA expression of inflammatory cytokines Tnfa and Il6 was up-regulated in LPS/D-gal-induced hapatitis , in which DNA fragmentation and activation of caspase-3 were subsequently observed .Immunoblot analysis showed that both mTOR pathway and NF-κB pathway were ac-tivated.Unexpectedly , inhibition of mTOR signaling could neither decrease the apoptosis in the liver nor increase the sur -vival of mice .Conclusions The results of the present study indicate that mTOR signaling may play pleiotropic roles in the pathogenesis of LPS/D-gal-induced hepatitis .

Objective By means of value-added reflection of students' learning behavior, this article gives an objective description of the current situation of pharmaceutical undergraduate education and puts forward some suggestions to strengthen the connotation construction of pharmaceutical education.Method Using the Chinese College Student Survey (CCSS) developed by Tsinghua University and Indiana University (US), 1100 students participated in a one-to-one web-based anonymous questionnaire, and using SPSS19.0 to analyze the data.Five comparable indexes and students' recognition behaviors were discussed using descriptive analysis and independent sample t test.Result The five comparable index scores and learning behavior skill index scores of the students in Chinese traditional medicine specialty and pharmacy were higher than the national overall level of all kinds of colleges and universities of science categories.Compared with the national norm of medical colleges, except that the score of 43.36 of the five comparable indexes in pharmaceutical professional academic challenge (level of academic challenge, LAC) was less than 43.63 national medical colleges LAC norm level, the other index scores were higher.LAC and active collaborative learning scores of Chinese pharmaceutical majors were lower than those of pharmacy students, and the difference was statistically significant (P＜0.05).In the study of behavioral skills, the scores of two major students' use, evaluation and comprehensive behavior were lower than the national standard of medical institutions, and there was a statistically significant difference in the scores of Chinese medicine and pharmaceutical majors (P＜ 0.05).Conclusion Teaching experience and learning support need to be optimized for pharmaceutical students.Learning scientificalness and comprehensive abilities in pharmaceutical education need to be improved.Mid and higher level of learning behavior needs to be enhanced for pharmaceutical students.

Objective To investigate the efficacy and safety of laparoscopic splenectomy (LS) in the treatment of immune throm-bocytopenia(ITP) .Methods 230 cases of ITP were collected in this hospital from December 2011 to December 2012 and randomly divided into two groups with 115 cases in each group .Among them ,the control group was treated by the conventional treatment of open splenectomy(OS) ,while the observation group was treated by LS .The differences of the operative time ,hospitalization dura-tion ,intraoperative blood loss and platelet count ,clinical effects and the occurrence situation of adverse reactions were compared be-tween the two kinds of operation method .Results The operative time and the length of hospital stay in the observation group were significantly shortened compared with the control group (t=14 .706 ,t=13 .625 ,P<0 .05);intraoperative blood loss was significant-ly decreased(t=18 .238 ,P<0 .05) and the platelet count was significantly increased (t=17 .248 ,P<0 .05);the total effective rate of the observation group was 96 .52% ,which was significantly higher than 86 .09% of the control group ,(χ2 =9 .35 ,P<0 .05);the adverse reaction rate and the recurrence rate in the observation group were 6 .09% and 6 .96% respectively ,which were significantly lower than 10 .43% and 19 .13% in the control group(χ2 =8 .35 ,χ2 =8 .75 ,P<0 .05) .Conclusion LS for treating immune throm-bocytopenia has definite efficacy with high effective rate ,low adverse reactions and low recurrence rate .

Objective To investigate the effects of Ulinastatin (UTI) on the expression of NF-κB and ET-1 in rats with acute necrotizing pancreatitis (ANP)-associated lung injury and morphology of lung tissue.Methods 60 Sprague-Dawley rats were randomly divided into 3 groups with 20 rats in each group,including sham operation (SO) group,ANP group and UTI group.ANP was induced by injection of 5% sodium taurocholate (1 ml/kg) into pancreatic duct;normal saline was injected for SO group with same amount.UTI was injected for UTI group with the amount of 10 000 U/L via tail vein after ANP induction.The rats were sacrificed 24 h later.The contents of serum amylase,TNF-α and wet/dry weight ratio of the lung were measured.The expression of NF-κB and ET-I protein were detected by immunohistochemical method.The level of apoptosis was detected by TUNEL Results The contents of serum amylase,TNF-α,and wet/dry weight ratio of the lungin in UTI group at 24 hours were (5 648±378)IU/L,( 89.19±3.54) ng/L and 4.55 ±0.07,respectively;which were significantly lower than the corresponding (6 799±437 )IU/L,(183.30±8.18) ng/L and 4.89±0.20 in ANP group (P＜0.05 ).There was no NF-κB and ET-1 expression,and no apoptosis was present in SO group.The positive rates of NF-κB and ET-1 in UTI group were (19±3 ) % and(8±1) %,respectively,the corresponding values in ANP group were (25±2) % and (13±1 ) %,respectively (P ＜ 0.05 ).The level of apoptotie index in UTI group was 13.75±1.25,which was higher than that (6.90±0,85) in ANP group ( P ＜ 0.05 ).Conclusions The high expression of NF-κB and ET-1 in lung tissue may cause lung injury.UTI could ameliorate the microcirculation and lung injury caused by inflammation.