ObjectiveTo explore the sensitivity of cancer stem cells to chemotherapy in human pancreatic carcinoma.MethodsPANC-1 ceils were cultured in an incubator filled with 5％ CO2 at a temperature of 37℃,and were labeled with Hoechst 33342.The SP analysis and sorting were performed using a FACSVantage SE.RT-PCR was performed to detect the expression of human CD133 ABCG2 and Notch1.SP and non-SP cells from the PANC-1 cell line were treated with 5-fluorouracil (5-FU; 1,10,or 100 μg/ml) or gemcitabine (10,100,or 1000μg/ml),and the cell viability was determined using the MTT assay.The sensitivity of sorted tumor cells to chemotherapeutics was determined in NOD/SCID mice model.ResultsSP cells were found to have higher drug-resistance both in vivo and in vitro and higher levels of mRNA expression for CD133,ABCG2 and Notch1,when compared to non-SP ceils.The xenografted tumors derived from injected SP cells and treated with gemcitabine had more CD133± cells than the untreated group.ConclusionsThe SP of PANC-1 pancreatic carcinoma cells are enriched with highly gemcitabine-resistant CSCs and determine the carcinogenesis of the PANC-1 pancreatic carcinoma cells.

Objecfive To establish the quality standard of Zaodi Anshen Pills. Methods TLC method was used to identify Spinosae and Radix polygoni Multiflori. HPLC method was used to determine the schisandrin, deoxyschizandrin and schisandrin B. Eclipse XDB-C18 column (250 mm×4.6 mm, 5 μm) was adopted by gradient elution using MeOH-water solution as the mobile phase. The detection wavelength was at 254 nm, column temperature was at 40 ℃ and the flow rate was 1.0 mL/min. Results The TLC spots were clear without interference of the negative control. The linear ranges of schisandrin, deoxyschizandrin and schisandrin B were within 338-1690 ng (r 2=0.999 1), 128-640 ng (r 2=0.999 3) and 236-1180 ng (r 2=0.999 4), and the average recoveries were 99.2%, 99.0% and 98.9% respectively. Conclusion This method is accurate, reliable, specific and reproducible, and can be used for quality control of Zaodi Anshen Pills.

Objective To evaluate emergency nursing methods to permanent pacemaker patients with severe complication and acute hemodynamic changes during implantation. Methods The emergency nursing methods were analyzed retrospectively in 9 of 2 027 patients over 6 years, who had serious complication occurred during the pacemaker implantation Results There were 4 patients with pneumothorax, 2 with pericardial tamponed, 1 with ventricular fibrillation and 2 with acute left ventricular failure in 9 patients (0.4%). Observed the changes of symptoms and signs carefully,found the severe complications at the first time and performed the specific nursing based on different kinds of conditions and improved the prognosis of these patients. Conclusions The serious complications during the implantation of permanent pacemaker were life-threatening, and the specific nursing scheme to different types of complications could increase the successful rate of emergency service.

Objective To observe the expression of connective tissue growth factor (CTGF) in pancreas, and discuss its significance. Methods The pancreatic fibrosis model was induced by high fat diets. The rats were sacrificed 16 weeks later, and the pancreatic tissue was harvested for routine pathologic examinations. Pancreatic collagen fibrosis I was determined by HE and Sirius red staining;α-SMA and CTGF expression were detected by immunohistochemistry. Results After pancreatic fibrosis, pancreatic lobules and acinar atrophy was observed, lobules gap was widened, interstitial fibrous tissue was significantly proliferated, the synthesis of pancreatic collagen fibrosis I was significantly increased when compared with normal pancreas ( 1500.2 + 255.8 vs. 57.4 ± 23.2, P ＜ 0. 01 ), the expression of α-SMA was significantly increased when compared with normal pancreas( 1500.2 + 255.8 vs. 57.4 + 23.2, P ＜ 0. 01 ), and the expression of CTGF was significantly increased when compared with normal pancreas (2950.5 ± 431.9 vs. 382.2 + 190.8, P ＜0.01 ), and there were abundant activated PSCs. Conclusions CTGF participated in the regulation of pancreatic fibrosis development; the function of CTGF was closely related to PSCs activation.

Objective To compare the efficacy of alfentanil and remifentanil in minimizing the propofol injection pain. Methods A total of 175 adult female patients undergoing gynecological procedures with general anesthesia were randomly divided into four groups.Patients received alfentanil 1mg(2 mL,AL group,n=43),remifentanil 0.01 mg(2 mL,REM1 group,n=43),remifentanil 0.02 mg(2 mL,REM2 group,n=45) or normal saline(2 mL,control group,n=44) 30 seconds prior to propofol administration.Visual analogue scale(VAS) was employed to evaluate the subjective feelings of pain due to propofol injection,and adverse effects were recorded. Results One patient in REM2 group and one patient in control group were excluded due to difficulty in venous catheterization.The injection pain in AL group,REM1 group and REM2 group was significantly less severe than that in control group(P

The degranulation of mast cells represents a pivotal e-vent in the allergic disorders.The Src family kinases(SFKs)are as a starting signal in the activation of mast cell.Lyn,Fyn and Syk play important regulatory role in the degranulation of mast cells.Regulating SFKs can reduce the degranlation process and inhibit the allergic disorders.Therefore,SFKs inhibitors can be potential drugs in the allergy.It is necessary to study the targeted medicine of SFKs,which will be a new direction of drug develop-ment.

Objective:To screen the proteins interacting with the human cytomegalovirus(HCMV)UL132 protein from the human fetus brain cDNA library by using Yeast Two-Hybrid System, and to elucidate the possible mechanism of UL132 protein in congenital cytomegalovirus infection.Methods:The HCMV UL132 fragment was amplified by polymerase chain reaction,the amplified HCMV UL132 fragment and expression vector pGBKT7 were digested and purified,and the HCMV UL132 fragment was linked to the vector pGBKT7.The pGBKT7-UL132 was constructed and transformed to yeast AH109, then the Human Fetal Brain DNA Library DNA was transformed into AH109 yeast.Using HCMV UL132 as abait, a human fetus brain cDNA was screened and the proteins interacting with UL132 protein were searched, the positive clone was sequenced and analyzed by bioinformatics methods.Results:The bait expression vector pGBKT7-UL132 was successfully constructed.The results of double enzyme digestion showed that there were two visible bands of 800 and 7 000 bp, respectively.After transformation of library plasmid, the transformation efficiency was calculated, and the transformation efficiency was 6.6×103 cfu· μg-1.There were 95 blue clones by X-gal coloration reactionsequencing and there were 10 clones interacting with the protein encoded by UL141 protein.The BLAST analysis showed that 7 of them were highly homologous with CAML.Conclusion:CAML might be one interaction protein with HCMV UL132 in Human Fetus Brain cDNA Library,suggesting that the interaction may be associated with the invasion and proliferation of the HCMV.

Objective To screen a human fetal brain cDNA library for proteins that can interact with HCMV UL145 using a yeast two-hybrid system.Methods A bait plasmid (pGBKT7-UL145) was constructed.Using HCMV UL145 as bait,a human fetal brain cDNA library was screened and proteins interacting with UL145 were identified using bioinformatic methods to sequence and analyze the positive clones.Results Three clones interacting with HCMV UL145 were found,and identified as FOXG1.Conclusion Several proteins interacting with HCMV UL145 in the human fetal brain cDNA library were identified as FOXG1,indicating that this protein may play an important role in the course of HCMV infection.

Fermentation conditions for the production of spores of Bacillus mucilaginosus mutant 021120 were optimized through the single factor and orthogonal experiments.The results showed that the biomass and formation of spores were obviously affected by carbonate,followed by nitrogen.Addition of CaCO3 and enhancing air flux notably promoted the formation of spores.The optimal culture medium was composed of 2% starch,0.4% Yeast,0.1% K2HPO4,0.1% MgSO4?7H20 and 0.5 %CaCO3,pH7.5.6% of the inoculum prepared with two-stage extensive culture was inoculated into a 70L fermentor.The fermentation was carried out with 2.0~2.5 vvm of air flux at 32℃ for 38~42h,and the spores of 9.80?108 cfu ml-1 was obtained.Under these optimal fermentation conditions,the capsule was successfully controled and the formation of the spores was effectively improved,thus the satisfying bacteial product was obtained.

Objective To analyze the epidemiological characteristics of overseas imported malaria reported in Chongqing City from 2011 to 2015,so as to provide the evidence for improving the imported malaria control strategies. Methods The epi-demiological data of overseas imported malaria cases were collected and analyzed descriptively for the species,original coun-tries,diagnosis and treatment in Chongqing City from 2011 to 2015. Results A total of 148 overseas imported malaria cases were reported in Chongqing City from 2011 to 2015,in which 96(65.54%)cases were falciparum malaria,and 37(24.32%) cases were vivax malaria;125(84.46%)cases were infected in Africa,and the rest of 23(15.54%)cases were infected in Southeast Asia. Most of the patients were 30 to 50 years old male workmen,and the ratio of male to female was 11.42:1. There was no obvious seasonal distribution among the reported timelines of the cases;however,there were two small peaks from June to August and from January to February. The median interval time of imported malaria cases from malaria onset to see a doctor was 1 day and from seeing the doctor to get malaria diagnosis was 2 days. The patients'first selected institutions were county medical institutions(50 cases,33.78%),then provincial medical institutions(36 cases,24.325%)and private doctors(20 cases,13.51%),and only 79(53.38%)patients got malaria diagnoses in their first selected institutions. The standard treatment were provided to 142(95.54%)cases. There were 43(29.05%)patients had serious complications and three patients were dead. Conclusion It is very important to enhance the multi-sector's collaboration to establish the collaborative investigation mechanism for screening malaria patients,and strengthen malaria health education for overseas workers and training courses in primary care medical institutions.