With the deepening of the national medical and health system reform,it is particularly important to establish the regional medical cooperative system with the large hospital as the core.The Affiliated Hospital of Xuzhou Medical University establishes interactive coordination relation with county-level hospitals through township hospitals in the cooperation model for collectivize operation,trusteeship and technical support,and it has accumulated some practical experience.The paper investigates and contrasts the core hospital and the member hospital development of the before and after the cooperative relationship,and sums up 6 key measures that the core hospital promoted the regional medical cooperative system construction,that is,improving the member hospital grade,strengthening talent development,support technology,specialty developments,hardware support and public project cooperation in order to provide references to promote the development of the regional medical cooperative system.

Objective To establish a method of quality control for compound Cornu cervi degelatinatum tablets. Methods Thin layer chromatography ( TLC ) was used for qualitative identification of Lonicerae Japonicae Caulis and Pyrola Herba in the preparations. High performance liquid chromatography was used to determine the content of loganin and chiratin in Lonicerae Japonicae Caulis and monotropein in Pyrola Herba. Results The TLC spots were clear and specific in the qualitative identification.HPLC determined the content. In the concentration range, monotropein, loganin and chiratin had a good linear relationship with peak area (r>0.999).The mean recovery was 97.2%, 98.4% and 96.0%;RSD was 1.4%, 0.8% and 0.8%. Conclusion The present study employs TLC for quality control and HPLC for quantity control. The methods are simple and accurate, have good reproducibility, and can effectively control the quality of compound Cornu cervi degelatinatum tablets.

Objective To study and evaluate the diagnostic and differencial diagnostic value of detection of human telomerase reverse transcriptase(hTERT) mRNA and telomerase activity in pancreatic juice of (pancreatic) cancer(PC) .Methods hTERT mRNA expression and telomerase activity in pancreatic juice in 24 cases of (pancreatic) cancer and 14 cases of chronic pancreatitis(CP) were detected and compared.Results Expression of hTERT mRNA was detected in 87 .5% (21/24) of PC cases and 21.4%(3/14) of CP cases(P

We studied the regulatory effects of the estragen receptorbeta (ERbeta) gene on the downstream estrogen signal transfection pathway in colon cancer cells and the possible mechanisms involved. A human ERbeta gene recombinant expression plasmid, pEGFP-C1-ERbeta, was constructed and transfected into the Caco-2 colon cancer cell line, a line with low ERbeta gene expression. The expression of ERbeta mRNA and protein was detected 72 h after transfection. RT-PCR was used to examine the expression levels of the progesterone recepror (PR) gene containing the classic estrogen response element (ERE), the C-fos oncogene containing the AP-1 site (a non-classical ER binding site), the epigenetic modifying genes, such as Dnmt1, Dnmt3a, Dnmt3b, and histone methyltransferase (HMT), and the human mismatch repair gene hMLH1. Methylation-specific PCR was used to detect the changes in the methylated sites of the CpG islands in the promoters of the ERbeta, PR, and C-fos genes. The results indicated that the human ERbeta gene recombinant expression plasmid pEGFP-C1-ERbeta was successfully constructed and transfected into Caco-2 cells. As compared with the control group, the mRNA and protein expression of ERbeta gene was increased significantly 72 h after the transfection of pEGFP-C1-ERbeta into the Caco-2 cells. As compared with the control group, the mRNA expression of the PR, C-fos, Dnmt3a and Dnmt3b genes was increased significantly 72 h after the transfection of pEGFP-C1-ERbeta into the Caco-2 cells, but the mRNA expression of the Dnmt1, HMT, and hMLH1 genes decreased significantly (P<0.05). As compared with the control group, different degrees of demethylation occurred in the promoters of the ERbeta, progesterone receptor (PR), and C-fos oncogene 72 h after the transfection of pEGFP-C1-ERbeta into the Caco-2 cells. The methylation index of the estrogen signal transfection pathway in Caco-2 cells was decreased significantly following the expression restoration of ERbeta gene (P<0.05). It is concluded that the restoration or up-regulation of the ERbeta gene in Caco-2 cells may significantly activate the expression of the related target genes in the downstream estrogen signal transfection pathway and may result in the demethylation changes of the pathway. During the process, the expression level and activity of the epigenetic modifying genes and the human mismatch repair gene have changed simultaneously. The regulatory effect of the ERbeta gene on the estrogen signal transfection pathway to a certain extent partly involves demethylation.

Objective To evaluate the clinical results of total laparoscopic hepatectomy for hepatolithiasis. Methods The clinical data of 72 patients with intrahepatic lithiasis receiving total laparoscopic hepatectomy in our hospital from July 2005 to April 2009 were retrospectively analyzed. Results The mean age of the 72 patients was (43. 8±21.7) yrs (16-65 yrs). For laparoscopic hepatectomy, it was anatomical left liver resection in 34 patients, anatomical resection of left lateral liver in 19 and resection of S6 in 16. The operative duration was (262.5± 115.5)min (125-320 min). The median intraoperative blood loss was 150 ml (50-400 ml). The occurring rate of postoperative complications was 12.50 %. Complications included bile duct infection in 8 patients, bile leakage in 6, gastroparesis in 1,postoperative early inflammatory ileus in 1 and subcapsular fluid collection of liver in 1. All the complications were cured by non-surgical means. Conclusion In the era of minimally invasive surgery, total laparoscopic hepatectomy has gradually become the prominent treatment for hepatolithiasis.

Objective To evaluate laparoscopic resection of gastric stromal tumors. Methods Clinical data of 20 patients undergoing laparoscopic resection of gastric stromal tumors from June 2003 to October 2007 were retrospectively analyzed. Result Laparoscopic wedge resection was completed successfully in all 20 patients with a mean operating time of(60±34) min, and without major complications. The mean hospital stay was (6.0±2.6) days. During a follow-up period from 10 to 22 months there was no recurrence. Conclusions Laparoscopic wedge resection is safe, effective, and minimally invasive for treating gastric stromal tumors.

Objective To investigate the feasibility and effectiveness of laparoscopic splenoctomy (LS)in patients with idiopathic thrombocytopenic purpura(ITP). Methods Clinical data of 17 ITP cases undergoing LS between Augest 2003 and December 2006 were analyzed retrospectively. Remits LS was Successfully conducted in all 17 cases without converting to open surgery with an average intraoperative blood loss of 120 ml in each case.There was no postoperative bleeding,fistula and infection.The platelet count increased rapidly in one week.After stopping glucocorticoid treatment for one month.15 cases achieved complete response(88.2%)and 2 caSes had partial response(11.8%).Fbllow-up of 3～43 months found no recurrence. Conclusions Use of LS for ITP is safe,feasible and effective.

Objective To evaluate the feasibility,safety and short-term outcome of laparoscopic-assisted surgery for colorectal cancer.Methods From August 2001 to November 2004,laparoscopic resection of colorectal carcinoma were performed in 112 cases,including right hemicolectomy(n=23),left himicolectomy(n=7),radical resection of sigmoid cancer(n=15),Dixon procedure(n=49),and Miles procedure(n=18).Results One hundred and five patients underwent laparoscopic resection successfully,7 cases were converted to open surgery because of hemorrhage,obesity or adhesion with adjacent organ,6 of which were left colon or rectal cancer.The mean operating time was(161.2?48.6)min,and the mean operative blood loss was 78.5 mL.There were 8 cases occurred postoperative complications,and no mortality during perioperative period.The length of upper and lower segment of resection for colonic cancer was (14.5?3.2)cm and(11.0?2.6)cm respectively.The length of upper and lower segment of resection for rectal cancer was(15.3?2.7)cm and(2.8?1.6)cm,respectively.The mean number of lymph nodes dissected was(8.2?4.6),and lymph node metastases were found in 49 cases.One hundred and seven cases(95.5%) were followed up for 8-44 months,of which,7 cases had local recurrence and 6 cases had distant metastases.No case of trocar port tumor implantation was observed.Conclusions Laparoscopic surgery for colorectal cancer is feasible and safe,can result in the same outcome as open radical surgery,and has the advantages of mini-invasive procedure.

Objective:To construct GST-tagged human NRP-1 fusion protein expression vector and induce its expression in Escherichia coli ( E.coli) ,then carry on inclusion body refolding and purification so as to obtain GST-NRP-1 fusion protein.Methods:NRP-1 gene was amplified by RT-PCR and inserted into pCR-blunt vector.Then the reconstructed plasmid was inserted into prokaryotic expression vector pGEX-4T-1.The constructed pGEX-4T-1-NRP-1 expression vector was transformed into BL21 cells and induced by i-sopropyl-β-D-thiogalactoside ( IPTG).Bacterial bodies were disrupted by sonication.Then the soluble fraction of fusion proteins were verified by Western blot and purified by Glutathione Sepharose 4B after inclusion body refolding.Results: The NRP-1 gene fragment was amplified by RT-PCR and inserted into pCR-blunt vector.Fusion protein expression vector pGEX-4T-1-NRP-1 was constructed suc-cessfully.After transformation, GST-NRP-1 expression vector was detected in BL21 cells and obtained purifying protein after refolding.Conclusion:The plasmid GST-NRP-1 was constructed successfully and laid basis for subsequent studies.

Objective This study aimed to establish the leukemia mouse model by using EL4/DsRed cell line expressing red fluorescent protein (DsRed) and to evaluate the model. Methods After total body irradiation with X-ray of 7.0 Gy, C57BL/6 mice were inoculated 5×106 bone marrow cells mixed different numbers of EL4/DsRed cells via tail vein. The model was evaluated by flow cytometry (FCM), reverse transcriptase-polymerase chain reaction (RT-PCR), and histopathology. Results The incidence of leukemia was 100 %. The presence of EL4/DsRed cells was found in liver, spleen, bone marrow and peripheral blood of recipients by FCM two weeks after transplantation. Pathological section revealed that all recipients had several organs infiltration apparently. With the increase in the number of inoculated tumor cells, the survival time of recipients was reduced and the infiltration of leukemia cells in organs was more serious. Conclusion Mouse leukemia model was successfully established when C57BL/6 mouse was intravenously transplanted with ≥5×102 EL4/DsRed cells. The model could be employed usefully in the future research such as the pathogenesis of leukemia and minimal residual disease (MRD).