Exosomes are secreted by living cells,which have the lipid bilayer and a large number of inclusions.We have found that exosomes derived from tumor cells play a crucial role in the development of tumor.Tumor-derived exosomes not only act as the messenger between tumor cells,but also perform as a carrier that affects the normal cells far away.Moreover,it has been reported to play positive role in the proliferation,differentiation,metastasis and immune resistance of tumor cells.

BACKGROUND: As a main gene engineering vector, adeno-associated virus (AAV) is characterized by its extensive host cells, lasting and stable expression and less immune response to hosts, and is applied widely. But AAV is a kind of defective virus, and need incasing cells to supply E1 protein. As important and special AAV incasing cells, AAV-293 cells can produce E1 in trans. But AAV-293 cells are delicated and cultivated difficultly, and the biological character is easy to be changed. Therefore, it is necessary to establish a culture method of AAV-293 cells to meet the need of gene engineering.OBJECTIVE: To establish a culture method of AAV-293 cells in vitro.DESIGN: An opening study.SETTING: Department of Neurology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: AAV-293 cells line was provided by Stratagene Corporation; high-carbohydrate OMEM (H-DMEM) powder by Gibco Company; there plasmids in AAV Helper-Free by Stratagene Company.METHODS: This experiment was carried out in the neurology laboratory of Tongji Hospital in Wuhan during the period from October 2006 to April 2007. AAV-293 cells were resuscitated and cultivated with H-DMEM growth medium in vitro, and were passaged and stored in liquid nitrogen when the cells monolayer confluence reached 50%. At the same time, their growing state was observed by inverted microscope, and their growth curve was noted. According to whether AAV-293 cells could give out green fluorescence or not (observed by fluorescence inverted microscope) after they were cotransfected with the there AAV system plasmids and infected with AAV supernatant, their biological character of packing AAV was assessed.MAIN OUTCOME MEASURES: ① Morphological observation of AAV-293 cells; ② the growth curve; ③ the package of AAV.RESULTS: ① AAV-293 cells observed by fluorescence inverted microscope were growing adhesively well with irregular polygons, light endochylemas and ambiguous nuclei appearances. ②The growth curve showed that the growing adaptive phase was the first day after AAV-293 cells were passaged, the actively growing phase was from the second day to the fifth day, and the growing platform phase was after the sixth day. ③ AAV-293 cells with green fluorescence observed by fluorescence inverted microscope, and cotransfection of the there AAV system plasmids was successful. AAV-293 cells gave out green fluorescence after infected with AAV supernatant, and AAV package succeeded.CONCLUSION: The culture method established by the authors in the experiment is simple and useful, and the cultured AAV-293 cells remain a good function of AAV package.

Objective To study preventive measures against conversions to open surgery during laparoscopic cholecystectomy(LC).Methods The study was based on 25 cases of conversions to laparotomy out of 852 cases of LC(2.93%)from December 1999 to October 2005 in this hospital.The clinical data revealed a bile duct injury in 3 cases,bleeding in 1 case,severe adhesion in abdominal cavity or between the gallbladder and neighboring tissues in 9 cases,disturbed anatomy and adhesion in the region of the Calot's triangle in 5 cases,cholecystoduodenal fistula in 2 cases,and Mirizzi syndrome(type Ⅱ)in 5 cases.Results The conversion to open surgery was successfully completed in all the 25 cases.No complication was encountered.Follow-up examinations for 1～2 years showed no bile duct stenosis.Conclusions Proper preoperative patient selection and use of intraoperative preventive measures are helpful to minimize the possibility of conversion to open surgery.

A 13 year old male farmer suffered from cutaneous and nail phaeohyphomycosis for five years is reported in this paper. The lesions were dull red, nodules and well defined plaques with ulcerations, mainly on the face, extremities, palmar and plantar surfaces and buttocks. Some of them were covered with thick greyish black crusts. General examination did not reveal abnormal findings except the skin lesions. Histopathology showed granulomatous response with numerous light brown septate branching hyphae. The colonies were greyish brown on SDA and PDA at 25℃ and 37℃ with brownish black ascomata. The ioslated strain was identified as Chaetomium globosum based on the morphological features of ascomata, ascomal hairs and ascospores.

Several years after trauma,a patien t suffered from skin verrucous hyperplasia for more than 30years,accompanied by ulcer occasio nally,on the feet,ankles and legs su ccessively.Mycological examinati on(9times)had been done before treatment.Slig ht septate hyphae and /or spores were demonstrated by direct mi-croscopy(5times);and yellowish green colony grew on S abouraud agar at 25℃for 2times,it wa s identified as Epidermophyton floccosum.Yeast -like colony grew on Sabourau d agar at 25℃for 5times,it was identi fied as Candida ciferrii by API.After treatment,Candida ciferrii could still be detected by mycologic al examination(6times),but no Epidermophyton floccosum was found.Slight septate hyphae were demonstrated in stratum spinosum,stratum granulosum and stratum corneum by histopathological examination.Isolated or clustered blastospores,spores and chlamydospores were demonstrated in stratum c orneum by histopathological examin ation.Therefore,it was concluded that the skin verrucous hyperplasia in this c ase is caused by mixed infection of Epidermophyton floccosum and Candida ciferrii.The patient was treated with flucon azole capsule 50mg daily for5weeks,followed by terbinafine tab let 250mg daily for 36weeks,and then250mg twice daily for 18weeks.In the 5th week of terbinafine therapy,a part of the lesions on the shin of left leg healed.No further therapeutic effect was observed.

Objective To study the correlation between postoperative tumor recurrence and VEGF expression in preoperative biopsy specimens of early gastric carcinoma together with p53 gene abnormality and tumor cell proliferation. Methods Sixty-eight endoscopically biopsied specimens obtained preoperativelly from patients with early gastric carcinoma were studied immunohistochemically. Date was processed by computer image analysis system. Result The rate of VEGF positivity was significantly higher in patients (n=5) with postoperative tumor recurrence than in those (n=63) without (5/5 vs. 14/63, P

Objective To understand the characteristics of common causes of acute pancreatitis (AP) in China and evaluate the association of the aetiology with gender and mortality.Methods The relevant literature was searched by the China journal full-text database (CNKI),Wanfang database,Weipu database and other databases and proceeding.Based on collecting data retrieval strategy,according to the inclusion criteria selection literature,Meta analysis was performed mainly for gallstone,hyperlipidemia,alcohol and other AP from the aspects of gender and case fatality rate.Results The Meta analysis included 11 articles which were accordance with the criteria,involving 13 601 patients,including 6 732 cases of biliary AP,1 372 cases of hyperlipidemia AP and 1 169 cases of alcohol AP.The Meta analysis showed that biliary AP male to female ratio was 0.79 ∶ 1,hyperlipidemic AP male to female ratio was 1.54 ∶ 1,alcoholic AP male to female ratio was 10.47 ∶ 1,overeating AP male to female ratio was 1.29 ∶ 1,and there was significant difference (P ＜ 0.05).Alcoholic AP mortality rate was the highest,which was 2.81 times than the biliary AP and 2.46 times than the hyperlipemic AP,and there was significant difference (P ＜0.05).Conclusions Biliary tract disease is the main etiologic cause of AP in China,and there are more females than males.The mortality rate of alcoholic pancreatitis is the highest,and there are more males.But we should investigate further high-quality,large-scale trails in patients with AP.

Objective To evaluate the indications and effectiveness of lower incisor extraction in orthodontics.Methods Twenty-three adult patients with lower incisor extraction were included in the study.The cases consisted of 14 female and 9 male patients with an average age of 23.2 years at the start.One single lower incisor was extracted in 17 patients and the other tow upper premolars were extracted in 6 patients.Cephalometric and casts analyses were carried out.Results Total treatment time was 13-23 months.All extraction space was closed after the treatment.All patients achieved satisfactory treatment results,with normal overbite and overjet and stable occlusal relationship.There were no significant changes before and after treatment on cephalometric analysis.The intercanine width was reduced by from pretreatment to posttreatment [(26.89--2.89) mm vs (23.92-t-1.54) mm (P＜0.05)].Conclusions The intercanine width is reduced after lower incisor extraction.In order to achieve satisfactory clinical effect,we should carefully select cases and pay attention to the corresponding techniques and methods.

Objective To investigate the association between hepatic fibrosis and anti-neutrophil cytoplasmic antibodies (ANCA)in the patients with schistosomiasis. Methods Totally 145 schistosomiasis patients confirmed by Jingzhou First Peo-ple's Hospital during the period of February 2014 to June 2016 were randomly selected as an experimental group,and 100 healthy people during the same period were randomly selected as a control group. The serum samples of the two groups were col-lected and the level of ANCA was detected by indirect immunofluorescence,and the levels of hyaluronic acid,laminin,procol-lagen type Ⅲ,and collagen type Ⅳ were detected by radioimmunoassay. The levels of the above indexes of the two groups were compared. Results The positive rate of ANCA was 28.96%(42/145)in the experimental group and 1.00%(1/100)in the healthy control group,and the difference between them was statistically significant(P < 0.05). The levels of hyaluronic acid, laminin,procollagen type Ⅲ and collagen type Ⅳ in the experimental group were significantly higher than those in the control group,and the differences were statistically significant(all P < 0.05),and the proportions of those with abnormal levels of hyal-uronic acid,laminin,procollagen type Ⅲ and collagen type IV in the experimental group were significantly higher than those in the control group,and the differences were statistically significant(all P < 0.05). In the experimental group,among the 42 pa-tients with ANCA positives,the proportions of those with abnormal levels of hyaluronic acid and laminin were significantly high-er than the proportions of those with ANCA negatives,and in the former,the levels of the four indicators of liver fibrosis were all higher than the normal values,and the average levels of hyaluronic acid and laminin in the patients with ANCA positives were significantly higher than those in the patients with ANCA negatives(all P < 0.05). Conclusion The patients with schistosomia-sis hepatic fibrosis could be positive for the autoimmune antibody ANCA,and ANCA may be associated with the autoimmune process of liver fibrosis.

OBJECTIVE To monitor the distrubution and the change in drug resistance of pathogenic bacteria from lower respiratory tract in patients with comprehensive intensive care to use the antibiotic properly in the clinics.METHODS All lower respiratory infection cases admited to the ICU in 2005-2006 were reviewed,and the distribution of pathogenic bactieria and their drug resistance profile were analyzed.RESULTS 89.3% pathogenic bacteria were Gram-negative ones,among them Pseudomonas aeruginosa(47.6%) and Klebsiella pneumoniae(21.2%) were prevailed.Staphylococcus aureus(6.2%) was the most prominent Gram-positive bacteria,MRSA showed increasing trend.Fungi accounted for 5.8%.All pathogenic bacteria showed high resistance to the antibiotics.CONCLUSIONS The pathogenic bacteria and their resistance to the antibiotics are highly changed at ICU.Intensive observations from the clinicians are recommanded so as to make the antibiotics effective.