Objective To observe the distributive characteristics of glial cell line-derived neurotrophic factor(GDNF)in brain tissue during cerebral ischemia-reperfusion in rats,and its role in ischemic brain damage.Methods The model of focal cerebral ischemia was made by occluding middle cerebral artery(MCA) for 2 h and reperfusion for 0.5～48 h, HE Staining was used to investigate the histological features of ischemic cerebral damage,the immunohistochemical method was used to observe the distributive characteristics of GDNF in brain tissue in rats. Results The focal ischemic areas (including preoptic area, striatum and cortex) presented at 0.5 h of reperfusion and peaked at 24 h. The neurons presented irreversible degeneration at 6 h of reperfusion. At 24 h, the ischemic area in the preoptic area developed into infarct form. At 0.5 h of reperfusion, neurons in ischemic cortex showed GDNF weak positive, neurons in peri-ischemic regions showed GDNF moderate positive. During reperfusion 3～48 h, neurons in ischemic regions showed GDNF negative. Up to 48 h of reperfusion, the active microglias or macroglias in periinfart area strongly expressed GDNF.The number of GDNF positive cells in all groups decreased (P

We present a minimal invasive technique for the treatment of broncho-pleural fistula (BPF) after pulmonary lobectomy with lung cancer. 2 cases of BPF were found at the 4th and 7th day after pulmonary lobectomy respectively. They were reoperated on by VATS, direct resuture of stump and consolidation with acrylic or fibrin glue. Both cases were cured. Authors consider mentioned procedure might be a feasible therapeutic approach for early broncho-pleural fistula.

Objective To study the clinical application of video-assisted thoracoscopic surgery (VATS). Methods Clinical records of 78 cases of VATS from July 1998 to December 2002 were retrospectively analyzed. There were 40 cases of bullectomy for spontaneous pneumothorax, 12 cases of surgical exploration for chest trauma, 9 cases of wedge resection for solitary pulmonary nodules, 6 cases of pleural biopsy combined with pleurodesis, 4 cases of resection for mediastinal tumor, 4 cases of pulmonary lobectomy, 2 cases of broncho-pleural fistula following lobectomy and 1 case of resection for esophageal leiomyoma. Results A conversion to open surgery was required in no cases, but a supplementary mini incision was needed in 5 cases because of the adhesion of pleural cupula. The intrathoracic drain was all removed within 48 postoperative hours with exception of 3 middle-old aged patients with spontaneous pneumothorax in whom the continuous air leakage was observed and the drain was removed on 7, 8 and 13 postoperative days, respectively. The delayed healing of drain site was seen in 5 cases. The incidence of operative complications was 10 3% (8/78). Conclusions Broad prospects exists in the clinical application of VATS, but the costs of the disposable is subject to reduce. The application of suture or knotting under thoracoscope may save medical costs.

Objective To explore clinical features of graft-versus-host disease (GVHD) after allogeneic non-myeloablative stem cell transplantation (NST) for haematologic diseases.Methods Eighteen patients were divided into three groups. Group A: Six severe aplastic anemia (SAA) adult patients underwent unrelated umbilical cord blood transplantation (UCBT). Group B: Combined transplantation of G-CSF primed allogeneic bone marrow and peripheral blood stem cells (PBSCs) was performed for 5 SAA patients. Group C: Seven malignant haematologic patients underwent transplantation of bone marrow cells for 3 patients or PBSCs for 4 patients. The protocol consisted of nonmyeloablative conditioning regimens based on anti-themocyte globulin (ATG) or anti-T-lymphocyte globulin (ALG). GVHD prophylaxis consisted of cyclosprine (CSA) and methylprednisolone (MP) for groups A and B, and methotrexate and CSA for group C. Mixed chimerism (MC) patients in group C were subjected to donor lymphocyte infusion (DLI).Results Four patients in group A achieved and sustained MC status, among them, one patient died of fungal septemia and one patient left hospital voluntarily. Three patients in group B achieved short period MC with donor chimerism more than 94 % at early stage post transplantation and converted into full donor chimerism (FDC) with long-term disease-free survival (DFS) and one patient developed chronic GVHD (cGVHD) 8 month post- trasplantation . Another two patients receiving donor stem cell infusion (DSI), one died of secondary mediastina lymphoma after 6 months and one patient recovered haematopoiesis. All patients achieved MC with haematologic partial remission (PR), and did not complicated acute GVHD (aGVHD) prior to DLI. Two cases died of severe infection and lost follow-up respectively. Another 5 patients gradually converted into FDC and achieved haematologic complete remission after 4, 3, 7, 5 and 4 DLIs, but they developed cGVHD (n=4), aGVHD (n=2) and myelosurppression (n=2).Conclusion The treatment of NST for SAA patients achieved better clinical effect with lower incidence of GVHD, and characterized by lower incidence of aGVHD and early mortality and higher incidence of cGVHD and infection for malignant haematologic diseases.

BACKGROUND: At present, non-viral vectors have become a hotspot in gene transfection research because of their strong points of lower toxicity,low immunologic reaction, target orientation, and easy assembly.OBJECTIVE: To evaluate the role of chitosan-DNA nanoparticles in transfection efficiency and cellular toxicity of tumor cells.DESIGN: Observational control trial. SETTING: Institute of Environmental Medical Sciences, Tongji Medical College of Huazhong University of Science and Technology.MATERIALS: Zeta potential/ particle size analyzer; spectrofluorometer;Hoechst 33258; PLPS-3'EGFP plasmid; lung cancer cell A549 and human hepatocarcinoma cell line HepG2.METHODS: This experiment was conducted in the Institute of Environmental Medical Sciences, Tongji Medical College of Huazhong University of Science and Technology, from December 2004 to December 2005. The green fluorescent protein gene was used as report gene; chitosan green fluorescent protein plasmid nanoparticles were prepared with re-coherence gy of prepared nanoparticles; Zeta potential/ particle size analyzer was used to measure the diameter and superficial potential of the nanoparticles;enzyme-protection test was used to measure anti DNA enzyme degradation and lung cancer cell A549 were transfected in vitro. Transfection of the cells was observed under the inverted fluorescence microscope after 24, 48nanoparticles.characteristics: Nucleic acid encapsulation efficiency of chitosan nanoparticles was 91.7%, and the nanoparticles presented globular shape with the mean diameter of 149nm and superficial potential of +20.5 mV.Transfection rate of nanoparticles: It reached the peak 48 hours later; the transfection rate of A549 was 95% while that of HepG2 was only about chitosan could inhibit the growth of HepG2 and A549, and the inhibitory effect of chitosan on cellular growth was stronger than that of the nanoparticles.CONCLUSION: Nanoparticles of chitosan plasmid can transfect HepG2and A549, two kinds of tumor cells, and have inhibitory effects on their growth, suggesting that nanoparticles, as the carrier of DNA, can be used in the transfection of tumor cells.

OBJECTIVE To investigate the drug resistance of Gram-negative bacteria in our hospital′s intensive care unit(ICU).It′s important to apply rationally antibiotics in clinic.METHODS We recruited all samples from infected patients of ICU during from Jan 2005 to Feb 2006.Totally 245 strains were identified to be Gram-negative bacteria.Kirby-Bauer method was performed to test extended spectrum ?-lactamases.RESULTS The first 3 of 245 strains were Pseudomonas aeruginosa(22.4%),Escherichia coli(20.8%) and Klebsiella pneumoniae(16.3%).Antibiogram analysis shew that CTX-M was the major type.CONCLUSIONS Imipenem keeps the highest sensitivity and antibacterial activity for Enterobacteriaceae and nonfermenters.Cefoperazone/sulbactam,cefepime,ceftazidime and amikacin keep the higher antibacterial activity.

AIM:Allogeneic hematopoietic stem cell transplantation for treatment of acute lymphoblastic leukemia characterized with high rate of relapse,especially in Ph+ patients.Presently,researchers focus on how to resolve relapse.Transplantation time,transplantation schedule and adoptive immunotherapy after transplantation are important.The study was performed to preliminarily observe treatment effectiveness after myeloablative stem cell transplantation,non-myeloablative transplantation after donor lymphocyte infusion and non-myeloablative transplantation followed by low-doses of Cyclosporin A.METHODS:Five patients were admitted at Department of Haematology of First People's Hospital between December 1998 and May 2007.Acute lymphoblastic leukemia patients were informed consent for allogeneic hematopoietic stem cell transplantation.The experiment was approved by hospital ethics committee.Among them,one patient was used the traditional preconditioning of busulfan and cyclophosphamide.Four cases were performed with non-myeloablative hematopoietic stem cell transplantation,and one of them was treated with reduced intensity regimen based on anti-thymocyte globulin donor lymphocyte infusion after transplantation;Three cases with fludarabine-based non-myeloablative transplantation were used low-dose Cyclosporin A after engraftment.Graft-versus-host disease prevention regimen was consisted of short-range methotrexate combined with Cyclosporin A.Haematopoiesis,chimerism,graft-versus-host disease and infection were observed after transplantation.RESULTS:All patients achieved successful engraftment.①One patient with mixed chimerism received eight donor lymphocyte infusion based on anti-thymocyte globulin-non-myeloablative transplantation and gradually converted into full donor chimerism with disease-free survival,and complicated acute graft-versus-host disease of skin and liver.②Three patients achieved full donor chimerism based on fuladarabine-non-myeloablative transplantation,one patient relapsed without graft-versus-host disease,and other two cases eliminated BCR/ABL fusion gene-positive cells with acute and chronic graft-versus-host disease.③One case after myeloablative transplantation relapsed and complicated with acute and chronic graft-versus-host disease.CONCLUSION:①The traditional,anti-thymocyte globulin or fludarabine-based non-myeloablative conditioning for transplantation in the treatment of adults with acute leukemia will be eligible for the successful implantation,and adoptive immunotherapy have graft-versus-leukemia effect.②The efficacy and complications of three transplantation strategies should be further studied.

Objective To evaluate the application of donor lymphocyte infusion (DLI) after nonmyeloablative stem cell transplantation (NSCT) for hematologic malignancies. Methods Donor lymphocyte infusion was performed on 5 hematologic malignant patients with mixed chimera (MC) and hemato logic partial remission of case 1, 2, 3, 4 or progression of case 5 after NSCT. The patients received the first DLI on the 4th to 5 th week posttransplant. The first T cell dose of ( 0.5 ～ 1.0 )?10 5/kg was followed by the escalated doses to the range of ( 0.5 ～ 2.0 )?10 8/kg. The total of procedures were performed at an average of 4.6 procedures (range 3～8) at intervals of 3～4 weeks. Results The MC was converted gradually into complete chimera (CC) after DLI in case 1, 2, 3 and 4 who were subjected to 7, 2, 3, 3 procedures respectively, and eventually converted into hematologic complete remission (CR), while case 5 remained MC and progression. The micro residual disease (MRD) of case 2 and 3 disappeared after DLI. Grade Ⅰ/Ⅱ acute graft versus host disease (aGVHD) in case 1, 2 and extensive/limited chronic graft versus host disease (cGVHD) in case 3 and 4 were found, and myelosuppression in case 2 and 4 was found as well. Conclusion Transient mixed donor recipient hematopoietic cell MC may be successfully converted into complete CC by DLI post transplant, and DLI can eliminate MRD. GVHD and myelosuppression remains major complications of DLI.

Eight patients with severe aplastic anemia received unrelated cord blood transplantation at the Guangzhou First People’s Hospital from June 1998 to December 2007. The patients were conditioned with decreased dosage of immunosuppressive agents of cyclophosphamide and antilymphocytic globulin. The median infused donor total nucleated cell were 5.69?10 7/kg of recipient weight, and the CD34+ cell was 4.10?105/kg of recipient weight. Methotrexate and corticoid methylprednisolone were used for prophylaxis of graft versus host disease (GVHD). The time to reach an absolute neutrophil count of 0.5?109/L ranged from 7 to 25 days (median: 17 days) and the time to reach a platelet count of 20?109/L ranged from 13 to 102 days (median: 35 days) after transplantation. DNA finger print map of 7 patients showed donor and recipient chimera. Two patients developed grade I to II acute GVHD, which was controlled. One patients developed grade II chronic GVHD, which was controlled by using methylprednisolone. Five patients had lived for 10-108 months, with no diseases. Taken together, unrelated umbilical cord blood transplantation is effective for adult patients. Partial conditioning regiment could ensure engraft of unrelated umbilical cord blood transplantation.

BACKGROUND:Natural colagen is considered to have low immunogenicity and good biocompatibility relatively. OBJECTIVE:To evaluate the immunogenicity of animal-based colagenin vitro. METHODS:Type I colagen was extracted from bovine tendon after immunogenicity removal. The colagen purity was detected by high performance liquid chromatography and residual DNA was measured quantitatively by fluorescence staining. Fifty BALB/c mice were randomly divided into five groups: subcutaneous injection of normal saline solution (negative control), bovine colagen (positive control), 33.4, 66.8, 133.4 mg/kg of bovine tendon colagen, respectively, once a day. After 12 days of continuously subcutaneous injection, lymphocyte proliferation, and cel classification and NK cel kiling function of mice were detected; after 3 weeks of continuous injection, the spleen, liver, spleen and lung tissue of mice were taken for histological examination. RESULTS AND CONCLUSION:Compared with the standard type I colagen, the purity of purified type I bovine colagen reached more than 99%, but the residual DNA was below 1 mg/L which was far less than the residue level of conventional cel-free DNA in the matrix (dry weight: 50-100 μg/g). After 12 days of continuous injection, there were no changes in lymphocyte proliferation, NK cel kiling function and the proportion of lymphocyte subsets. After 3 weeks of injection, the spleen and lymph sheath of mice around the smal artery became thickened in the 66.8 and 133.6 mg/kg bovine tendon colagen groups, which could cause accidental liver injury and lung injury, but the splenic corpuscle germinal center area had no change. These findings indicate that continuously subcutaneous injection of animal-based colagen can cause the lower lymphocyte immune response to the spleen of BALB/c mice, which may cause accidental liver and lung injuries.