AIM: To determine the ultimate preservation time of murine cardiac grafts in 4℃ Ringer's solution. METHODS: Murine cardiac grafts were implanted to the abdominal vessels heterotopically 2, 4, 6, 8 and 10 hours after cold preservation. Graft survival rate and histological morphological changes, as well as the neutrilphil, T cell, macrophage infiltration, ICAM expression were determined by immunohistochemistry. RESULTS: The cardiac function-recovery rate and 1-week graft survival rate were 100% and 83.3% in 6-hour preservation group. Compared with non-preservation control group, no more apparent histological damages, cell infiltration and ICAM expression were found. CONCLUSION: The ultimate preservation time of murine cardiac graft in 4℃ Ringer's solution was 6 hours. [

Objective To investigate the clinical significance of detecting mammaglobin in sentinel lymph nodes(SLN) of breast cancer. Methods In 32 breast cancer patients, methylene blue was injected around parechyma of breast cancer to locate the SLNs, and nested RT PCR(reverse transcriptase polgmecane chain reaction) was used to examine the expression of mammaglobin mRNA. Results The SLN was successfully identified in 30 of the 32 cases(93.8%).The micrometastases detection between group SLNs and non SLNs had significant statistical difference(P

ObjectiveTo study the expression of pleiotrophin (PTN) in colorectal cancer and its impact on the malignancy.MethodsRT PCR and immunoperoxidase staining were applied on the resected samples.ResultsPTN mRNA was expressed in 18 cases of 24 colorectal cancer tissues and 18 cases of 19 adjacent normal tissues. PTN protein was also found expressed in stroma, but the expression rate was higher in colorectal cancer tissues (34%) than in adjacent normal tissues (9%, P

AIM: To test the effects of FTY720 on mouse intestinal allografts.METHODS: C_3H mice(H-2~k)were used as donor and C57BL/6 mice(H-2~b) as recipients.FTY720 group,allogeneic control group and isogeneic control group were set up.6 and 14 days after transplantation,murine intestinal grafts were harvested for histologic assessment.Lymphocytes were collected from mesenteric lymph nodes(MLN),Peyer's patch(PP),lamina propria lymphocytes(LPL) and intraepithelial lymphocytes(IEL) in the graft,then were analyzed by cytometry.RESULTS: Rejection was inhibited in FTY720 group at the 6th post-transplant day,although not at the 14th day.Recipient CD4~+ and CD8~+ T cells,CD19~+ B cells,as well as ?? TCR lymphocytes,were greatly reduced by FTY720 therapy.The similar action of FTY720 was also revealed in Gr1~+CD11b~+ monocytes.CONCLUSION: FTY720 is efficient on alleviating allo-immune response by reducing the infiltration of both lymphocytes and monocytes into the graft in a mouse intestinal transplantation model.

Objective: To investigate hyperthermia enhanced expression of heat shock protein70 (HSP70) in breast cancer drug sensitive cell line MCF7 and multi-drug resistant (MDR) cell line MCF7/ADR, so as to increase cytotoxic activity of immunologic effector cells against the target cells, and to provide an experimental basis for cell immunotherapy based on hyperthermia. Methods: The immunological effector cells were induced and expanded by cytokines. Breast cancer cell lines MCF7 and MCF7/ADR were treated at 42 ℃ for an hour. Then after being incubated for additional 4 hours, 24 hours and 48 hours, the cells were digested. Flow cytometry (FCM) was used to detect the expression of HSP70 on the target cells. MTT assay was employed to evaluate cell proliferation and the cytotoxic activity of immunologic effector cells against target cells. Results: The expressions of HSP70 in both the target cells had significant difference. The expression of HSP70 in MCF7/ADR cells was higher than in MCF7 cells before hyperthermia. After hyperthermia the expression of HSP70 increased by 6 folds in MCF7/ADR cells, and 22 folds in MCF7 cells and was higher than in MCF7/ADR cells at hour 4. The proliferation inhibition fraction of hyperthermia against MCF7 was significantly lower than that of MCF7/ADR. The cytotoxic activity of immunologic effector cells against MCF7 cells was lower than against MCF7/ADR cells before hyperthermia. After hyperthermia the cytotoxic activity of immunologic effector cells against MCF7 cells rose by 86.23%, and was higher than against MCF7/ADR cells (rose by 30.32%). Conclusion: The expression of HSP70 in breast cancer drug sensitive cell line MCF7 and MDR cell line MCF7/ADR were enhanced significantly by hyperthermia. Cytotoxic activity of immunologic effector cells against both the target cells were increased by hyperthermia. The differential expressions of HSP70 in breast cancer drug sensitive cell line MCF7 and MDR cell line MCF7/ADR affect the cytotoxic activity of immunologic effector cells.

Objective To investigate the clinical efficacy of neoadjuvant chemotherapy combined with radical gastrectomy for advanced gastric cancer.Methods The retrospective cross-sectional study was conducted.The clinicopathological data of 73 patients who underwent neoadjuvant chemotherapy combined with radical gastrectomy for advanced gastric cancer at the First Affiliated Hospital of Zhejiang University between June 2004 and December 2009 were collected.Neoadjuvant chemotherapy regimens included XELOX and FOLFOX.Patients received radical gastrectomy within 2 weeks after the completion of the last cycle of neoadjuvant chemotherapy and then continued to undergo postoperative neoadjuvant chemotherapy.Observation indicators:(1) adverse event of neoadjuvant chemotherapy;(2) surgical and postoperative situations;(3) follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect survival of patients up to December 2014.Measurement data with skewed distribution were described as M (range).Overall survival time was from the beginning of treatment to death or end of follow-up (patients with loss to follow-up).Progression-free survival time was from the beginning of treatment to tumor progression,recurrence and metastasis or death.The survival curve was drawn by the Kaplan-Meier method.Results (1) Adverse event of neoadjuvant chemotherapy:of 73 patients,38 received XELOX regimens and 35 received FOLFOX regimens,with a median cycle of 3 (range,1-7 cycles).There were 55 adverse events during neoadjuvant chemotherapy,including 47 with grade 1-2 and 8 with grade 3-4.(2) Surgical and postoperative situations:all the 73 patients underwent successful D2 radical gastrectomy for gastric cancer,including 40 receiving total gastrectomy,31 receiving distal gastrectomy,1 receiving total gastrectomy with transverse colon resection and 1 receiving distal gastrectomy with cholecystectomy.Of 73 patients,10 with postoperative complications were improved by conservative treatment,including 3 with pleural effusion,2 with peritoneal effusion,2 with anastomotic bleeding,2 with cholecystitis and 1 with lympha fistula.No patient received reoperations or died within 30 days postoperatively.Pathological TNM staging:22 patients were detected in stage Ⅰ-Ⅱ,45 in stage Ⅲ,4 in stage Ⅳ and 2 in stage T0N1M0.Three patients (in stage T0N0M0) had complete remission.Forty-three patients underwent postoperative chemotherapy.(3) Followup:all the 73 patients were followed up for 8-125 months,with a median time of 51 months.The median survival time,5-year overall survival rate and 5-year disease-free survival rate of 73 patients were 52 months,41.1％ and 34.2％,respectively.Conclusion XELOX and FOLFOX regimens of neoadjuvant chemotherapy combined with radical gastrectomy for advanced gastric cancer are safe and effective.

AIM: To establish a method of in vitro donor heart perfusion in murine cardiac transplantation during preservation and apply it in adenovirus mediated gene transfection for donor heart. METHODS: Donor heart was transfected with recombinant adenovirus and stored for 2 hours after harvest, then it was transplanted heterotopically in abdomen. The grafts were appraisal by palpitation. Marked gene products were determined by X-Gal staining, aod T cell infiltration was determined by immunohistochemistry. The activation markers of recipients' lymphocytes were examed by cytometry. RESULTS: The grafts survival rate is 100% after perfusion and cold storage. The LacZ staining became strong 1 week after transplantation. The grafts remained an intact structure and no apparent T cell infiltration. The activation status of recipients' lymphocytes were not enhanced by transfected cardiac graft. CONCLUSION: In vitro perfusion during graft cold preservation is feasible for adenovirus mediated gene transfection. [

OBJECTIVE To study the clinical features of acute leukemia(AL) patients with complication of infection. METHODS A retrospective analysis of prevalence of infection occurring in 123 AL patients,their bacterial spectrum,and the effect of G-CSF on the infection were done. RESULTS The prevalence of infection in AL patients was 94.3%,among which 49.2% were infected with Gram-positive organisms,42.4% with Gram-negative bacilli and 8.4% with fungi.Blood culture occurred mostly with Escherichia coli and Pseudomonas infection.Patient′s neutropenia was significantly related to the infection.The patients with neutrophil count less than 0.2?10~9/L had more frequently suffered severe infection with long-lasting time,and had more than two sites of infection.Age,hemoglobin level,prophylactic intestinal usage of antibiotics could not reduce patient′s infection.The total mortality of AL patients with infection was 6%.Pulmonary infection and septicemia increased mortality,but G-CSF therapy reduced it. CONCLUSIONS AL patients are at high risk of infection which is significantly associated with severe neutropenia.G-CSF therapy exerts an assistant role to antibiotics in controlling the infection.

Objective To detect EPPB41L3 methylation frequency difference between esophageal squamous cell carcinoma (ESCC) tissues and the normal tissues and between ESCC patients′plasma and healthy volenteers′plasma, and to analyze the correlation with clinicopathological parameters. Methods We collected esophageal squamous cell carcinoma tissues (n = 42 patients) and adjacent surrounding normal tissues (n = 42 patients), and plasma from 42 patients with ESCC and from 50 healthy individuals. We used methylation specific PCR (MSP) combined with agarose gel electrophoresis to detect the methylation status of the EPB41L3. We used the SPSS 13.0 software for statistical analysis by χ2 test and Fisher′s exact test. Results EPB41L3 frequency of methylation was significantly higher in tumor tissues than in the adjacent tissues (59.5% vs. 4.8%), the difference was statistically significant (χ2 = 28.873, P < 0.001). For plasma, EPB41L3 methylation frequency was 31.0%in cancer patients, while was not detectable in the healthy volunteers. Methylation of EPB41L3 in tissues was more frequently found in patients with tumor size of ≥ 5 cm or T3 than in patients with tumor size of < 5 cm or T1-2. Conclusions The methylation frequency of EPB41L3 is higher in ESCC tissues than in control normal tissues, and higer in plasma from ESCC patients than that from the healthy volunteers. EPB41L3 methylation is more frequently found in patients with more advanced disease.

Grid management plays an important role in improving the efficiency of social service management. The authors explored the establishment of a nosocomial infection grid management model and implemented it at a tertiary general hospital in Shanghai. χ2 test results showed that since the hospital fully implemented the grid management model in early 2016, the number of hospital infections, surgical site infections, type I surgical incision infections, and ventilator-related pneumonia had shown a downward trend year by year. The number of inspections before the use of antibacterial drugs, the number of blood cultures submitted for fever patients, and the timing of hand hygiene compliance showed an increasing trend year by year( P<0.05), and the management of nosocomial infection was more effective than the previous period. The application of grid management model required establishment of grid organization structure, control of key links, implementation of the special personnel responsibility, formulation of a performance evaluation system, and implementation of incentive measures. The grid management model can significantly reduce the risk of hospital infection, improve the efficiency of hospital infection management, and was valuable to be applicated and promoted in medical institutions.