Objective To enrich pancreatic cancer stem cells through culturing mammospheres, and to detect the expressions of epidermal growth factor receptor(EGFR) and a disintegrin and metalloprotease 9 (ADAM9) and investigate their significance. Methods PANC1 cells were cultured in serum-free conditioned medium to continuously generate mammospheres, and parts of mammospheres were cultured on a collagen substratum to induce differentiation. Mammospheres cells and differentiated cells were collected, flow cytometry was used to detect the proportion of side population (SP) cells, and the expressions of EGFR, ADAM9 mRNA and protein were detected by real-time PCR and Western blotting. Results PANC1 cells mammospheres were successfully generated and could be passed continuously. After differentiation, mammospheres cells could regain the ability of adherent growth. The proportion SP cells in mammospheres cells and differentiated cells were ( 5.40 ± 0.38 ) % and (2.80 ± 0.42 ) %, and the difference was statistically significant ( P ＜ 0. 05 ).Compared with differentiated cells, the expression of EGFR and ADAM9 mRNA of mammospheres cells upregulated 2.5 and 3.0 folds ( P ＜ 0. 05 ). The expressions of EGFR and ADAM9 protein of mammospheres cells were 0.90 ± 0. 09 and 0.64 ± 0.07, which were significantly higher than those in differentiated cells (0.62 ±0.11 and 0.48 ±0.09, P ＜0.05). Conclusions Mammosphere cells contained higher proportion of pancreatic cancer stem cells. ADAM 9 may play an important role in the occurrence and development of pancreatic cancer through the EGFR signaling pathway.

According to the requirement of simulated actual combat,based on the traditional contents of sanitary reconnaissance of water source,this paper introduced some practical knowledge and skills in drinking water under emergency,including water source search,utilization,drinking principles and some measures to reduce the loss of body water.These practices enriched and consum- mated the subject of sanitary reconnaissance of water source.

Objective To explore the effect of preoperative chemotherapy for children with malignant extragonadial germinoma. Methods Twenty patients with malignant extrogonadal germinoma had been treated with preoperative chemotherapy from Mar.2003 to Mar.2007.Protocol 1 of chemotherapy was to improve PEB:(cisplatin plus etoposide plus bleomycin A5);Protocol 2 was VAC(vincristine plus actinomycin D plus cyclophosphamide) the average time span of the chemotherapy was 16 weeks.Upon completion of chemotherapy,tumor resection was performed.The content of alpha fetoprotein(AFP) was determined before operation and compared with the content before chemotherapy.Re-gular follow-up procedure was taken after operation. Results All 20 patients received chemotherapy and underwent operation.The removal rate of resection operation was 100%,the process of operation was smooth.The volume of tumors in 14 patients decreased over 50% compared with those before chemotherapy.The response rate was 70%.The content of AFP decreased significantly in 15 patients(75%).Fifteen patients had been living without tumor for up to 2 years,for 5 cases the survival time went beyond 5 years. Conclusions Malignant extrogonadal ger-minoma in children grow infiltratively,bring detrimental effects to neighboring organs,and bring obstacles to resection operation.Chemotherapy for children with malignant extragonadal germinoma before operation can reduce volume of tumor and bleeding during operation,raise the resectability rate and lower the content of AFP.It will enhance the effectiveness of operation and improve prognosis.Therefore,it can be used as a conventional clinical method.

Cerebellum ; surgery ; Cystadenoma, Papillary ; complications ; pathology ; surgery ; Epididymis ; pathology ; surgery ; Follow-Up Studies ; Genital Neoplasms, Male ; complications ; pathology ; surgery ; Humans ; Keratin-7 ; metabolism ; Kidney ; surgery ; Male ; Middle Aged ; Mucin-1 ; metabolism ; Vimentin ; metabolism ; von Hippel-Lindau Disease ; complications ; metabolism ; pathology ; surgery

Apoptosis ; drug effects ; Caspases ; physiology ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; pathology ; Membrane Potentials ; drug effects ; Mitochondria ; drug effects ; physiology ; Plant Extracts ; pharmacology ; Proanthocyanidins ; pharmacology ; Seeds ; chemistry ; Vitis ; chemistry

Acupuncture Points ; Adolescent ; Adult ; Aged ; Asthma ; therapy ; Combined Modality Therapy ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Treatment Outcome ; Young Adult

Objective To investigate the genotypes and encoding resistance genes differences of Acinetobacter baumannii and analyze their interrelations with multi-drug resistance.Methods A total of 77strains Acinetobacter baumannii were collected random from the second Xiangya Hospital during September 2008 to September 2009.The K-B method which was WHO recommended was adopted to Acinetobacter baumannii drug sensitivity test to 15 kinds of antibiotics to establish susceptibility spectrum.At the same time,random amplified polymorphic DNA(RAPD)technique was used to establish DNA fingerprinting.The genes of β-lactamase(TEM-1,IMP,OXA-23,OXA-24,AmpC),aminoglycoside-modifying enzymes[aac(3')-Ⅰ ,aac(6')-Ⅰ ,ant(3")-Ⅰ]and 16S rRNA methylase(armA,rmtA,rmtB)were detected by PCR and sequenced,and find the relationship between the gene encoding and multi-drug resistance.In addition,we compared the rates of resistance genes of Acinetobacter baumannii and the relations with the genotype and the multi-resistance.Results Thirty-one sensitive strains and 46 multi-drug resistance strains(10 Pan-drug resistances)were isolated.Seventeen types from A to Q were separated using RAPD technique.E genotype widely popular in the ICU was the advantage type in multi-drug resistance strains,and the rate was 47.1%.While the various types scattered in sensitive strains.The positive rates of TEM-1,IMP,OXA-23,OXA-24,AmpC,aac(3')-Ⅰ ,aac(6')-Ⅰ ,ant(3")-Ⅰ ,armA in the multi-drug resistance strains and the sensitive strains were 95.7%,39.1%,84.8% ,54.3%,87.0%,89.1%,84.8%,45.7%,63.0% and 58.1%,9.7%,32.3%,48.4%,48.4%,29.0%,45.2%,12.9%,9.7%,respectively,and there was significant difference except for OXA-24 using the X2 test(P ＜ 0.05).All isolates were negative for rmtA gene and rmtB gene.Drug susceptibility analysis showed that the resistant rate was significantly higher of the strains carrying resistant genes than that of the resistance negative strains.When the strains were resistant to gentamicin and amikacin,the rate of three aminoglycoside genes positive was 34.8%.The trains containing all the measured β-lactamase genes were all resistant strains.Conclusion Compared with the sensitive Acinetobacter baumannii strains,a broad resistance spectrum and a high drug resistance rate were showed in multidrug resistance strains isolated from clinic,which harboring many kinds of β-lactamase genes and aminoglycosides genes with a high separation rate,and the same clone of multiple drug-resistant strains may be transmitted in and among wards.

Objective To investigate the prevalence of 16S rRNA methylase gene armA and to analyze their effect on the drug resistance in multi drug-resistant strains of Acinetobacter baumannii . Methods A total of 72 Acinetobacter baumannii isolates were collected from the Second Xiangya Hospital from Jan. 2008 to Dec. 2008. The size of inhibitory zone of these strains to gentamycin, tobramycin and amikacin were determinate using Kirby-Bauer( K-B) method. The 16S rRNA methylase genes armA were detected by PCR. PCR products were purified and sequenced. Then we used randomly amplified polymorphic DNA method (RAPD) genotyping technology for the establishment of DNA fingerprinting. In addition, we compared drug sensitivity test with RAPD technology. Results Twenty isolates of 72 strains were armA positive and the resistance rates of the strains with armA gene to gentamycin, tobramycin, amikacin were 90.0% , 90.0% and 90. 0% , respectivily. armA positive stains were divided into 7 types using RAPD technology. A genotype was the advantage type. Conclusion The study showed that 16S rRNA methylases gene armA was prevalent in Acinetobacter baumannii which could lead to resistant to almost all aminoglycosides at a high level. And the main form of armA gene prevalence in our hospital was the spread of the same clone strain inside and outside of clinic department.

Objective To detect the variation of vascular endothelial growth factor (VEGF) in the splanchnic vessels under portal hypertension (PHT) and explore its mechanism and influence on the process of hyperdynamic circulation.Methods In humans,the level of VEGF pathway related proteins were detected in the splenic artery of PHT patients in clinical trials.In animal experiments,the following parameters were observed for rats in the control group (group N,n =7) and the CCl4 induced portal hypertension group (group PHT,n=7):portal vein diameter,portal vein blood flow velocity (PBV),portal vein blood flow (PBF),portal vein pressure (PP),norepinephrine (NE) reactivity in the isolated mesenteric artery microcirculation,contractile reactivity and degree of endothelial ni tric oxide synthase (eNOS) activation in the mesenteric artery by selectively inhibiting the VEGF signal pathway with SU5416.In cell experiments,primary culturing of arterial endothelial cells in vitro were used to verify the effects of VEGF on eNOS activation.Results The results showed that VEGF expression levels in the splenic artery of PHT patients significantly increased.In animal experiments,there was not a significant difference in portal vein diameter between group N and group PHT.How ever,the PBV and PBF of group PHT were lower than those of group N,and SU5416 had no clear effect on PBV and PBF in group PHT.PP of group PHT was much higher than that of group N,and SU5416 had little influence on reducing PP in group PHT.The contractile response of mesenteric artery microcirculation to NE in group PHT decreased significantly,EC50 increased a lot,and SU5416 improved this hypoergia partially.The protein levels of VEGF,VEGFR-2,eNOS,and p-eNOS in the mesentery artery of group PHT raised quite a lot compared to group N,and SU5416 decreased the protein level of VEGFR-2 and activation of eNOS significantly.Experiments in vitro confirmed that VEGF could promote the activation of eNOS.Conclusion The excessive VEGF produced in visceral arteries under PHT may participate in the process of hyperdynamic circulation partially through promoting the synthesis and activation of eNOS and then reducing visceral arteries' response to NE.

Objective To study the effect of chest wall vibration therapy on bronchiolitis. Methods A total of 64 patients with bronchiolitis were divided into an experimental group and a control group, the former included 34 cases and the latter included 30 cases. The experimental group received both routine treatment and chest wall vi- bration, while the control group only received routine treatment. PaO_2, PaCO_2, SaO_2, Heart Rate (HR) and Respi- ration (R) were observed, respectively, in the experimental group and the control group at the beginning and the end of the third day. Time needed for expectoration and length of hospital stay in the two groups were observed. Results It was shown that PaO_2, PaCO_2, SaO_2 , HR, R were significantly improved at the end of the third day when compared with those at the beginning in both groups(P