AimTo explore the effects of bcl-2 antisense oligonucleotide targeting the coding region of the bcl-2 mRNA on apoptosis induced by Vp16 in HL60 and K562 cells. Methods Drug sensitivity was compared by MTT cytotoxicity assay, and expression of bcl-2 protein and apototic cells were assayed by flow cytometry. Results The bcl-2 antisense oligonucleotide of 10 μmol · L- 1 combined with etoposide inhibited expression of bcl-2 protein,increased apoptosis in HL60 and K562 cells and decreased IC50 of etoposide; The antisense oligonucleotide targeting the coding region of the bcl-2 mRNA had stronger effection than the antisense oligonucleotide targeting the translation initiation. ConclusionThe bcl-2 antisense oligonucleotide targeting the coding region of the bcl-2 mRNA can enhances etoposide induced apoptosis of HL60 and K562 cell.

AIM The antisense drug design will be optimized based on bcl 2 mRNA secondary structure simulated with computer. METHODS bcl 2 mRNA second structures were simulated with computer and Mfold software, and the unstable zones on the second structure, as designing antisense zones, were selected. RESULTS Five antisense deoxynucletides were studied and evaluated with experiments of HL 60 and K562 leukemic cells. Two of them exist significant effect of inhibiting grow of HL 60 and K562 leukemic cells with dose of 10 ?mol?L -1 or more. CONCLUSION The designs with computer and corresponding software will be usefully efficient way to look for antisense drugs.

OBJECTIVE:To approach the information of stem cell transplantation in treating myocardial infarction.DATA SOURCES: An online search of Medline database was undertaken to identify articles about stem cell transplantation in treating myocardial infarction published in English from January 1980 to December 2005 using the keywords of "stem cells, myocerdial infarction, cell transplantation".STUDY SELECTION: The literatures were checked primarily, and articles about the stem cell transplantation in treating myocardial infarction were selected, and the repetitive studies and reviews were excluded.DATA EXTRACTION: Totally 106 relevant articles were collected, finally 30 of them were involved and the other 76 were excluded.DATA SYNTHESIS: Myocardial infarction occurs when the arteries that supply blood to the heart muscle (the coronary arteries) become hardened and narrowed. The transplantation of exogenous cells into damaged myocardium, a procedure known as cellular cardiomyoplasty (CCM), can be used to ameliorate the remodeling process by the regeneration of cardiomyocytes and replenishment of the vascular supply. Advances in stem cell therapy now provide wide cell sources for the treatment of cardiovascular disease by CCM such as skeletal myoblasts (satellite cells),embryonic stem cells (ESCs) and bone marrow-derived mesenchymal stem cells (MSCs). Although there are many problems to be resolved, recent in vitro and in vivo animal studies as well as clinical trials have suggested the promising potential of the transplantation of stem cells for the treatment of myocardial infarction.CONCLUSION:Stem cell transplantation acts as a more potential treatment measure for myocardial infarction. There are still some fundamental issues to be addressed for the clinical application in future.

OBJECTIVE: To investigate theresearch condition of mechanism of cardiomyocytes differentiation from embryonic stem (ES) cells and the application potency of ES cells at home and abroad.DATA SOURCES: We searched in Medline database from 1997 to 2005 with the key words of "embryonic stem cell,cardiomyocyte/cardiac myocyte,differentiation" in English.STUDY SELECTION: The retrieved data were selected primarily. Inclusive criterion: articles of differentiation into cardiomyocytes from ES cells.Exclusive criterion: the articles of repetitive research were excluded.DATA EXTRACTION: Totally more than 200 articles were researched,and 26 were included.DATA SYNTHESIS: Some articles of differentiation of ES cells into cardiomyocytes from the construction of primarily experimental methods of cardiomyocytes differentiation from ES cells to electrophysiologic study of ES cells-derived cardiomyocytes in monoplast level were collected. Furthermore, many investigators studied the expression of transcription factors that controlled cardio development in the differentiation of ES cells to cardiomyocytes from the angle of transcription factors. Cardiac transcription factors GATA4, MEF2C and NKx2.5 had been reported to control a cardiac gene program and thus to play a crucial role in transcriptional regulation during cardiogenesis. Meanwhile, calcineurin, MAPKs and other signaling pathways were also involved in the adjustment of cardiomyocyte differentiation. In addition, the purity of differentiated ES-derived cardiomyocyte cultures for myocardial infarction was a key issue.CONCLUSION: When ES cells are cultured within Ebs and in the absence of leukemia inhibitory factor (LIF), some of them can differentiate into cardiomyocytes. ES-derived cardiomyocytes express characteristic contractile proteins, such as MHC and myosin light chain (MLC)-2V. Transcription factors, including GATA4, MEF2C and NKx2.5, play a crucial role in the cardioblast differentiation and trigger the expression of specific genes such as α-actin, MHC and MLC2V. However, t he molecular mechanisms of cardiomyocyte differentiation from ES cells remain largely unexplored. The identification of factors that can promote cardiac differentiation is fundament in order to study cardiac differentiation.

Object To explore the protoplast fusion of Dendrobium candidum Wall ex. Lindl. and Gynostemma pentaphyllum. (Thunb) Makino. Methods The two kinds of protoplasts were fused by PEG method, and then cultured in the modified liquid medium containing 2 mg/L BA and 1 mg/L NAA. Results High yield, viability, and pure mesophyll protoplasts were isolated from D. candidum and G. pentaphyllum. Conclusion The first cell division occurres within three days after fusion. And some of the cells are divided three times.

Objective To explore the efficacy of radiofrequency ablation (RFA) combined with splenectomy in patients with small hepatocellular carcinoma (sHCC) associated with hypersplenism.Metheds The data of 100 patients with sHCC associated with hypersplenism who received RFA or hepatectomy combined with splenectomy were analyzed retrospectively.The patients were divided into the observation group and the control group based on the intraoperative approach.Fifty-three patients who received RFA and splenectomy were in the observation group,and the remaining 47 patients who received hepatectomy and splenectomy were in the control group.Multiple intraoperative and postoperative factors were compared between the two groups.Results There were significant differences between the two groups in warm ischemia time,operation time,intraoperative blood loss,hospital stay,and amount of blood transfusion (P ＜ 0.05).The postoperative complication rate of the observation group (7.6％,4/53) was significantly lower than the control group (44.7 ％,21/47) (P ＜ 0.05).There were no significant differences in 1-,3-,and 5-year survival (respectively,100.0％,75.5％,and 67.9％ vs 97.9％,76.6％,and 68.1％) and in disease free survival (96.2％,57.5％,and 41.7％ vs 93.5％,58.3％,and 43.8％ respectively) between the two groups (P ＞ 0.05).Conclusion RFA combined with splenectomy can be considered as an alternative treatment for patients with sHCC associated with hypersplenism.

Objective To analyze the causes of clinical misdiagnosis of solid pseudopapillary tumor of pancreas (SPTP),and recommend the countermeasures with a combination of pertinent literature.Methods The clinical data of 10 cases of SPTP with misdiagnosis were retrospectively analyzed.There were 9 women and 1 man,and the average age was 29.3 years old.The clinical symptoms were nonspecific.Results All patients received surgical treatment,and the correct diagnosis was made according to the pathology after surgery.All patients were followed up,and no recurrence or metastasis was found.Conclusions SPTP is a rare disease in clinic.Sufficient understanding in the clinical features and imaging characteristics can improve the rate of diagnosis before operation.

Objective To evaluate the effects of isoflurane anesthesia on the cognitive function and expression of translocator protein 18 kDa ( TSPO) in the brain tissues of aged rats. Methods Twelve pathogen?free male Sprage?Dawley rats, aged 20 months, weighing 500-550 g, were randomly divided into 2 groups ( n=6 each) using a random number table: control group ( group C) and isoflurane anesthesia group ( group I) . The rats inhaled 2% isoflurane in 100% O2 for 4 h in group I, or 100% O2 for 4 h in group C. The rats underwent Morris water maze test at 24 h after anethesia. The escape latency and frequency of crossing the original platform were recorded. Then the rats were sacrificed, and the hippocampus and cerebral cortex were isolated for determination of the expression of TSPO and Iba1 by Western blot and quantitative real?time reverse transcriptase?polymerase chain reaction. Results Compared with group C, the escape latency was significantly prolonged, the frequency of crossing the original platform was decreased, and the expression of TSPO and Iba1 protein and mRNA in the hippocampus and cerebral cortex was up?regulated in group I ( P<0. 05 ) . Conclusion Isoflurane anesthesia results in decreased cognitive function through up?regulating the expression of TSPO in the brain tissues of aged rats.

Objective To investigate the effects of gene transfection with human growth/differentiation factor 5(hGDF5)on the differentiation and proliferation of rabbit bone marrow mesenchymal stem cells (BMSCs).Methods BMSCs were obtained from adult New zealand rabbits and purified by gradient centrifuge.Exogenous recombinant human GDF5 was transfected into BMSCs with liposome method.Then hGDF5 expression at mRNA and protein level was measured separately by reverse transcription-polymerase reaction and indirect immunofluorescence method.Activity of alkaline phosphates(ALP),expression of TypeⅡcollagen(ColⅡ),proteoglycan and growth of the cells were all measured by biological methods to evaluate the effects of hGDF5 gene transfer on the differentiation and proliferation of rabbit BMSCs.Results After hGDF5 gene transfection,BMSCs expressed hGDF5 mRNA and protein,and compared with the control groups,expression of proteoglycan and ColⅡ increased significantly,but no significant difference appeared in ALP activity and cell proliferation.Conclusion Gene transfer with hGDF5 is an effective way to enhance the expression of GDF5 at mRNA and protein.The expression of heterogenetic hGDF5 gene can induce BMSCs' differentiation to chondrogenic cells.But the gene transfection has no obvious effects on the proliferation and ALP activity of BMSCs.

AIM: To investigate whether the bcl-2 antisense oligonucleotide increases the sensitivity of HL60 and K562 cell lines to daunorubicin. METHODS: IC50 for HL60 and K562 was determined with MTT method, the expression levels of Bcl-2 protein were assayed by immunofluorescence using fluoresce isothiocyanate labeling. In addition, apoptosis was detected by morphological observation and flow cytometric analysis of DNA fragmentation. RESULTS: It was found that the two oligonucleotides directed against the coding region and the translation initiation of bcl-2 mRNA, combined respectively with daunorubicin, inhibited expression of bcl-2 protein, increased apoptosis in HL60 and K562 cells, and decreased IC50 of daunorubicin significantly ( P