Purpose:To investigate effect on bcl-2 and bax expression in apoptic PGCl_(3) cells and to study the mechanism of apoptosis of PGCl_(3) cells induced by death associated protein kinase(DAPK).Methods:Eukaryotic express vector pcDNA3.1-DAPK was tranfected into High-metastasis Non-small Lung Cancer Cell PGCl_(3).Changes of bcl2 and bax expression were detected with RT-PCR.Phospharation of Bcl-2 was examined with Westernblotting.Results:It was demonstrated that bcl-2 expression was down regulated,but bax expression did not change in apoptosis of PGCl_(3) cells induced by DAPK.Level of Phospharation of Bcl-2 increased.Conclusions:Apoptosis of PGCl_(3) cells induced by over-expression of DAPK may be associated with down regulating of bcl-2 expressio and increase of phospharation of Bcl-2.

Objective To review the experience of surgical management for caustic esophageal stricture and discuss the surgical techniques of transverse colon interposition for esophageal replacement. Methods 106 patients with caustic esophageal stricture were treated surgically. 32 patients underwent colon interposition with colonphargageal anastomosis and others received colon interposition with a cerrical anastomosis. The ascending branch of the left artery of the transverse colon was preserved as the supporting vessel of the interpositioned colon. Results There was no postoperative death. The leakage of cervical anastomosis was observed in 12 patients, anastomosis stenosis in 8 patients, and tracheotomy was performed in 3 patients. All patients were perfectly recovered by the treatment. Conclusion Transverse colon interposition for esophageal reconstruction is an optimal approach in the treatment of caustic esophageal stricture.

10.3969/j.issn.2095-4344.2013.26.006

This paper describes the construction and practical experience of the key laboratory for teaching of biochemistry and molecular biology,and indicates that the laboratory promotes the development of teaching and scientific research.It is proved to be a suitable measure for sharing teaching resource,improving teaching quality and raising teacher' academic level.

Both Chinese GB nutrient agar and American FDA standard method agar are commonly used for counting bacteria in food inspection. A number of our experiments showed that the count of bacteria by FDA standard method agar is 23.9% higher than that by GB nutrient agar. The present C8 medium is an improved medium based on the composition of the two media mentioned above The result of our comparison experiment showed that the C8 medium used for counting bacteria was 35.8% higher than that by GB nutrient agar and 9.5% higher than that by FDA standard method agar respectively.

AIM: Open reading frame(ORF) of death associa ted protein kinase1(DAPK1) gene was cloned for studying on tumor forming and met astasis.METHODS: Based on nucleotide sequence of DAPK1 gene f rom GenBank, a pair of primers was designed. DAPK1 gene ORF was transfected into Raji cells in expression vector pcDNA3.1(+) with lipofectamine reagent. Morphol ogic assessment of apoptosis was performed with fluorescence microscope cytotoxi city and cell viability was assayed by MTT. RESULTS: DAPK1 gene ORF was amprified from K562 cells by RT-PCR. It was cloned into plasmid pMD18-T and sequenced. There were seven mutation in 4 300 bp nucleotide sequence rel ativel y to DAPK1 nucleotide sequence from GenBank, but six was synonymous mutation and one was single nucleotide polymorphism. 4 300 bp nucleotide of DAPK1 gene O RF was transfected into Raji cells. DAPK1 gene expression was detected in 48 h a fter it was transfected into Raji cells. Then Raji cells showed apoptosis.CONCLUS ION: Large fragment gene was cloned by RT-PCR and transfected into Raji cells successfully. Over-expression of DAPK1 gene induced Raji cells apoptosis.

BACKGROUND:Bone marrow stem cells are defined by their multi-potential ability, and can be differentiated into intrinsic cells in the kidney.
OBJECTIVE:To study the effects of mobilizing autologous bone marrow stem cells by granulocyte colony-stimulating factor plus stem cellfactor on cellapoptosis and proliferation of rats with renal ischemia-reperfusion injury.
METHODS:Total y 160 male Sprague-Dawley rats were randomly divided into four groups:control group, model group, cytokine treatment group, cytokine control group. Rat models of unilateral renal ischemia-reperfusion injury were established in the model and cytokine treatment groups. Rats in the cytokine treatment group and cytokine control group received subcutaneous injection of granulocyte colony-stimulating factor (50μg/kg) and stem cellfactor (200μg/kg), once a day, for 5 continuous days. Rats in the model and control groups had no treatment. Apoptotic cells were detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling method, and the expression of CD34-positive cells, Caspase-3, Bcl-2, proliferating cellnuclear antigen in the kidney were measured using immunohistochemistry staining.
RESULTS AND CONCLUSION:The number of CD34-positive cells in renal tissue of the cytokine treatment group was significantly higher than that of the control group and model group (P<0.05). The apoptotic index and expression of Capase-3 in the model group and cytokine treatment group were higher than those in the control group and cytokine control group (P<0.05). The apoptotic index and expression of Capase-3 in the cytokine treatment group were lower than that in the model group (P<0.05). The expression of Bcl-2 in the model group and cytokine treatment group was higher than that in the control group and cytokine control group (P<0.05). The expression of Bcl-2 in the cytokine treatment group was higher than that in the model group (P<0.05);however, as time went on, Bcl-2 expression was obviously decreased. Proliferating cellnuclear antigen expressed both in the model group and in the cytokine treatment group. Additional y, the proliferative index reached peak at 24 days in the model group, and then decreased gradual y;while in the cytokine treatment group, it reached the peak at 10 days, maintained a high level until the 17th day, and then decreased gradual y. Mobilization of autologous bone marrow stem cells by combination of granulocyte colony-stimulating factor and stem cellfactor can increase proliferation and decrease apoptosis of renal tubular epithelial cells after renal ischemia-reperfusion injury, and thus, promote the recovery from renal tubular injury.

Objective To explore the cell metabolism of the bilateral hippocampus effected by the inflammatory factor of blood serum in the development of the depression.Methods Using Proton magnetic resonance spectroscopy 1 H-MRS technology to detect multiple metabolic indices of the bilateral hippocampus of 20 first episode depression patients and 20 healthy persons,and detecting the levels of IL-6,IL-2,IL-10 in blood serum were detected.Finally the correlation between them was analyzed.Results Compared with the healthy group,the values of Glx/Cr and NAA/ Cr were reduced in left hippocampus (Glx/Cr: (0.82±0.48),t=2.69,P＜0.05 ; NAA/Cr: (1.12 ±0.44),t =2.81,P＜ 0.05) and the value of Cho/ Cr was increased in left hippocampus ((2.49± 0.78),t =2.36,P＜0.05),but only the value of Glx/ Cr were reduced in right hippocampus ((0.84 ± 0.47),t =2.43,P＜ 0.05),and the other metabolic indices changed unobvious.The depressive group had the significantly higher levels of IL-6,IL-2 in blood serum(IL-6: (12.47±3.19) ng/L,t=4.53,P＜0.05 ; IL-2: (29.44±5.72) ng/L,t=2.44,P＜0.05),but no significant difference was found in the IL-10 level in blood serum.The Glx/ Cr of bilateral hippocampus level was negatively correlated to the IL-6 and IL-2 level in blood serum(left:(IL-6: r=-0.555,P＜0.05;IL-2: r=-0.624,P＜0.05) ;right:(IL-6: r=-0.575,P＜0.05; IL-2: r=-0.523,P＜0.05)).The NAA/ Cr of left hippocampus level was negatively correlated to the IL-6 and IL-2 level in blood serum(IL-6: r=-0.582,P＜0.05;IL-2: r=-0.607,P＜0.05).The Cho/Cr of left hippocampus level was negatively correlated to the IL-6 and IL-2 level in blood serum(IL-6: r=0.601,P＜0.05; IL-2: r=0.552,P＜0.05).Conclusion The abnormal of glutamic acid system in bilateral hippocampus maybe the important performance and the potential originating link,the changes of the inflammatory cytokines level maybe the factor of these abnormal changes.

Objective To explore the clinical effect of percutaneous osteoplasty with SKy bone expander system in the treatment of calcaneal fracture.Methods 63 feet of 55 patients with calcaneal fractures were involved in this study.According to Sanders fracture classification including 38 feet of Sanders Ⅱ,18 feet of Sanders Ⅲ,7 feet of Sanders Ⅳ.There were 6 to 16 days interval between the injuries and the surgeries.The patients were treated by percutaneous osteoplasty with the SKy bone expander system.The standard of operation was the satisfaction of reduction and Bohler's and Gissane's angles under X-ray.Results All of 63 feet of 55 patients were followed up for average 22 months.According to the criterion of therapeutic effect,the results were as follows:excellent in 30 cases,good in 26 cases,fair in 7cases,and no poor case.The excellent and good rate was 88.9％.Conclusion Percutaneous osteoplasty with SKy bone expander system in the treatment of calcaneal fracture,especially in reduction and fixation of Sanders type Ⅱ and Sanders type Ⅲ,can recover Bohler's and Gissane's angles,significantly shorten the duration of illness,and has fast recovery and can possess satisfactory curative effect,and it is worth popularizing.

Objective To investigate the effect of cystatin C (Cys C) on adventitia in rabbit abdominal aorta restenosis after angioplasty and its mechanism.Methods 48 New Zealand white rabbits were randomly divided into injury group (receiving balloon dilation of abdominal aorta),the treatment group (taking Cys C monoclonal antibody therapy) and the control group (receiving femoral artery puncture and catheter sheath without balloon dilation and intervention of Cys C monoclonal antibody injection),and each group had 16 rabbits.Peripheral vein blood was drawn to measure the serum level of cystatin C before and 8 h,1 day,1 week,3 weeks,6 weeks after the operation in all rabbits.After 6 weeks of operation,the abdominal aorta were taken and stained with HE.Vascular morphometry analysis and adventitial cell count were conducted.Smooth muscle actin (SM-actin) and proliferating cell nuclear antigen (PCNA) expressions in the adventitia were observed by immunohistochemical staining.The number of PCNA positive cell in the adventitia was counted and the PCNA proliferation index was calculated.The vascular remodeling index,vascular external elastic lamina area (EELA),internal elastic lamina area (IEIA) were used to evaluate the vascular remodeling and the residual stenosis and vascular cavity area was used to measure the vascular stenosis.Results Plasma Cys C level began to rise at 8h after operation and reached the peak at 1 week after operation,and continuously increased for 5 weeks in injury group,and reached to respectively at 3 weeks and 6 weeks after operation.The Cys C levels were significantly higher in injury group than in the treatment and control groups at different time points (all P＜0.05).There were no significant differences in Cys C levels at different time points between the treatment group and the control group.The injury group showed that the number of PCNA positive cells was higher in injury group than in treatment and control groups,both P＜0.05).Compared with the control group,the vascular luminal area,EELA and IELA were significantly increased (all P＜0.05).After treated with the monoclonal antibody Cys C intervention,the treatment group showed that lumen area,vascular EELA,IELA was significantly decreased (P＜0.05),and the vascular remodeling index and residual stenosis rate were decreased as compared with the injury group (0.871 vs.0.784,33.1％ vs.19.7 ％,both P＜0.05).Correlation analysis showed that Cystatin C level was positively correlated with the vascular lumen area,neointimal area,internal elastic lamina area,external elastic lamina area and the number of PCNA positive cells (r=0.812,0.797,0.876,0.932 and 0.822 respectively,all P＜0.01).Conclusions Plasma Cys C level is increased in rabbit after abdominal aorta balloon injury and has a positive correlation with the severity of arterial stenosis.High Cys C level can induce adventitial fibroblast activation,proliferation,phenotype transformation and migration,and accelerate the processes of atherosclerosis and stenosis.Cys C level is the independent risk factor for abdominal aortic stenosis.