Apoptosis ; Apoptosis Regulatory Proteins ; genetics ; Genetic Vectors ; Humans ; K562 Cells ; Transfection

According to the limited class hours and requirements of medicinary major curriculum application,this paper attempts to make some choice of physical chemistry teaching contents and emphasize on the first class,and also to explore bilingual teaching of partial chapters and experimental teaching.

Objective To evaluate the therapeutic effects of autolngous bone marrow mesenchymal stem cells transplantation in the early phase of acute renal ischemia-reperfusion(I/R)injury in rabbits and their impact on tubular epithelial cell apoptosis.Method Bone marrow mesenchymal stem cells(MSCs)were isolated and cultured in vitro.Thirty rabbits were divided randomly into transplanted group,control group and sham-operated group,10 in each group.Rabbits of transplanted group were induced acute renal I/R injury by clamping bilateral renal arteries for 90 minutes,then transplanted MSCs labeled with BrdU immediately after the resumption of circulation in kidney.The rabbits of control group were induced acute renal I/R injury and then infused with saline instead.The sharn operated rabbits were transplanted autologous MSCs labeled with BrdU after sham operation.Fortyeight hours after transplantation,all rabbits were sacrificed.Renal functional and structural damage were evaluated.MSCs derived BrdU positive cells were determined by immunostalning.Tubular epithelial cell apoptosis was arialyzed by usign TUNEL.The SPSS version 11.3 Software was used for statistical analysis.ANOVA were used to ana lyze the data of renal function and tubular epithelial cell apoptesis.Data of renal structure darnage were analyzed by using the nonparametric Kruskal-Wallis test.Results Compared with control group,the renal functional and struc tural darnage were significantly ameliorated(24 h,BUN:F=7.483,Ser:F:15.091;48 h,BUN:F:17.741.Scr:F=61.865;P<0.05),tubular epithelial cell apoptods Wsa reduced significantly in transplanted group(F=135.495,P<0.01).MSCs derived BrdU positive cell were detected in the renal tissues of transplanted rabbits.Conelusiu Autologous bone marrow mesenchymal stern cells transplantation is curative for acute renal I/R injury in rabbits.Bone marrow MSCs might ameliorate renal I/R injury by reducing tubular epithelial cell apoptosis.

Objective To investigate haemodynamic change during hemodialysis and analyze effects of cardiac index (CI) on hemodynamic parameters and associated influencing factors in maintenance hemodialysis (MHD) patients.Methods Seventy-five patients bearing an arteriovenous fistula (AVF) entered the study.Cardiac output (CO),cardiac index (CI),central blood volume (CBV) and peripheral vascular resistance (PR) were determined by ultrasound dilution technique at the end of 1 hour,2 hours and 3 hours of dialysis.AVF blood flow (Qa) was also measured with the same device before haemodynamic parameters investigation.Results Mean age of patients was (55.84 ±12.39) years old (range 21-81 years) and 43 patients (57.3％) were female.Systolic arterial pressure,SV,CO,CI and CBV were significantly declined and no significant change for diastolic arterial pressure and heart rates at the end of 2 hour and 3 hour hemodialysis,whereas PR was increased gradually during hemodialysis.Patients were divided into there groups with CI less than 2.5 L·min 1·(m2)-1,2.5-4.2 L·min-1·(m2)-1 and more than 4.2 L·min-1 ·(m2)-1 at the end of 1 hour dialysis.Statistically significant decreasing SV,CO and increasing PR were detected in patients with CI＜2.5 L·min-1· (m2)-1 and 2.5-4.2 L· min-1· (m2)-1,compared with CI＞4.2 L·min-1· (m2)-1 group (P＜0.01).The hemodynamic change was the most obvious in the group of CI less than 2.5 L· min-1· (m2) 1,and no significant changes happened in CI＞4.2 L·min-1 ·(m2)-1 group.Some factors were found to be associated to CI values.Qa and systolic arterial pressure had positive relationship with CI,while age and diabetes had negative relationship with CI.Conclusions Systolic arterial pressure,CO,CI and CBV decrease and PR increases during hemodialysis.Obvious change occurs when CI is less than 2.5 L·min-1· (m2)-1.CI is associated with Qa,systolic arterial pressure,age and diabetes.

· AIM: To discuss the cause and management of complications during the manual small incision cataract surgery (MSICS).combined with intraocular lens implant (MSICS-IOL) were done in 160 patietns with cataract. In this paper, we reviewed the clinical data of the intraoperative complications.injury, posterior capsule rupture, iris prolapse and hyphema, were mainly caused by inexpert skill and lacking of experience.techniques, careful operation could decrease the incidence of complications during MSICS operation. By proper management, the intraoperative complications could be solved satisfactorily.

Objective The interleukin-1β (IL-13) expression in serum of acute myeloid leukemia (AML) patients was evaluated. To explore the significance of IL-1β in leukostasis and tissue infiltration by leukemic cells. Methods ELISA was used to investigate the contents of IL-1β in serum of 83 newly diagnosed with AML which contains 16 hyperleukocytic AML patients, and compared the IL-1β level between the hyperleukocytic AML group and non-hyperleukocytic AML group, the infiltrated group and non-infiltrated group. Results The content of IL-1β in AML serum [(88.23±36.30) pg/ml] was higher than that of in the control group[(29.56±15.53) pg/ml], with significant difference (P ＜0.01). There was a higher level for IL-1β in hyperleukocytic AML group[(136.67±65.68) pg/ml] than in non-hyperleukocytic AML group [(69.85±48.35) pg/ml],and there was a significant difference. IL-13 and peripheral blood cells were in linear correlation (r=0.74, P ＜0.01). There was a higher level for IL-1β in infiltrated group[(111.31 ±57.35) pg/ml] than in the other group [(79.68±43.42) pg/ml], and there was a significant difference. Conclusion The IL-1β may play an important role in leukostasis and tissue infiltration by leukemic cells in AML.

Objective To investigate the value of DNA-image cyt-ometry (DNA-ICM) in the diagnosis of urothelial cell carcinomas (UCC). Methods Totally 162 voided urine specimens (92 cases from urothel-ial car-cinomas patients and 70 cases from benign urinary system diseases patients ) were detected with DNA-ICM and liquid-based cytology (LBC), respectively. Results The sensitivity and specificity of DNA-ICM were 65.2%and 100% respectively in the diagnosis of UCC but those of LBC were 27.2% and 98.6%, respectively. The sensitivity of DNA-ICM was significantly higher than that of LBC in the diagnosis of UCC (P < 0.01). The sen-sitivity of DNA-ICM in upper urinary tract urothelial cell carcinomas (UTUC) were 77.1%, which was much higher than that in bladder urothelial cell carcinomas (57.9%) but no statistical significance was found (P >0.01). Conclusion DNA-ICM, which improves the positive rate of urinary cytology, has great application value in the diagnosis of urothelial cell carcinomas and it is an effective screening method for urothelial cancer in diag-nosis and follow-up.

Objective To construct the recombinant adeno-associated virus vector carrying cancerous inhibitor of PP2A (CIP2A)short hairpin RNA (shRNA)for preparation of high-titer viruses.Methods The small hairpin RNA of CIP2A (CIP2A shRNA)was designed,synthesized and cloned into pDC31 6-EGFP-U6 plasmid which was double digested by Bam HⅠ and Hin dⅢ.The resultant plasmid pDC31 6-EGFP-shRNA was confirmed and served as template to appraise primers.EGFP-CIP2A shRNA sequence was amplified by PCR,double digested with Eco RⅠand Sal Ⅰ and ligated to pSNAV2.0 plasmid digested with the same enzyme pair.pSNAV2.0-EGFP-CIP2A shRNA plasmid DNA was prepared,purified,identified and transfected into BHK-21 cells.BHK-21 cells expressing CIP2A shRNA (BHK-21/CIP2A-shRNA ) were obtained and subsequently infected with VGTC’s proprietary AAV packaging system to package the rAAV2-CIP2A shRNA.After purification,the functional and infectious virus was obtained and the titer of virus was detected.Real-time PCR and Western blot methods were used to detect the expression of CIP2A after infection with HepG2 cells,and the empty viral vector rAAV2-EGFP was used as control. Results A recombinant adeno-associated virus-2 vector carrying CIP2A shRNA was constructed successfully.The presence of the target sequence in the vector was confirmed by double enzyme digestion and sequencing.By transfecting the pSNAV2.0-EGFP-CIP2A shRNA plasmid into BHK-21 cells,BHK-21/CIP2A shRNA cells were infected with helper virus HSV1-rc/ΔUL2 to package the rAAV2-CIP2A shRNA to obtain a functional and infectious virus.The titer of the recombinant virus was 0.25×10 1 2 v.g./mL.The expression of CIP2A mRNA and screening value of 1×10 5 MOI effected HepG2 cells.Conclusion A high-titer recombinant adeno-associated virus-2 vector carrying CIP2A shRNA has been constructed successfully.

Objective To evaluate the clinical value of microvascular anastomotic devices in the reconstruction of lower extremity defects by free flap transfer.Methods A retrospective review of 42 consecutive patients who underwent lower extremity microvascular reconstruction performed from May 2013 to November 2013 by microvascular anastomotic devices was performed.Patient charts were reviewed for age,sex,etiology of defect,location of defect,flap type,anastomotic technique,complications and flap survival.Results No patients had an arterial or venous anastomosis revised.The vascular anastomosis patency rates was 100％ and the flap survival rate was 97.6％.Total complication rate (9.5％) was due to 1 partial flap loss,1 partial skin graft loss and 2 hematoma.There were no intraoperative or perioperative complications involving the use of a microvascular anastomotic device itself.Conclusion Microvascular anastomotic devices have effective vessel anastomoses in lower extremity microvascular reconstruction.Thus,it presents an ideal tool for lower extremity microsurgical reconstruction.

Objective To observe the expression of chemokine (MCP-1) and chemokine receptor (CCR2) in bone marrow cells,bone marrow stromal cells of multiple myeloma (MM) patients.Methods 15 cases were diagnosed by domestic uniform standard for MM patients,7 cases of male,8 cases of female,age range from 38 to 67 years,mean age 53.7 years old.According to the Durie-Salmon staging system,patients were divided into Ⅰ (2 cases),Ⅱ (5 cases) and Ⅲ period(8 cases).Control group were from 10 cases of non-malignant blood disease patients.MCP-1,CCR2 expression were measured by flow cytometry.Results Almost 14 cases of bone marrow cells expressed MCP-1and CCR2 in MM patients,while in the control group,bone marrow cells almost did not express MCP-1and CCR2.Stromal cells had similar MCP-1and CCR2 expression profile (68.17 ％ vs 4.27 ％. P＜0.05).Tumor cells of MCP-1/CCR2 expression rates were 3.25 ％ and 32.76 ％. Compared MCP-1/ CCR2 expression of stromal cells and tumor cells with different stages of disease, the activated stage and the stable stage had similar level (68.71％ and 32.76 ％ vs 70.12 ％ and 53.39 ％. P＞0.05). Conclusion Most patients with MM bone marrow were expressed MCP-1and CCR2.MCP-1and CCR2 are the major MM cell surface expression of chemokine/receptor, which play important roles in the progress of.