Objective To investigate the effects of mutated PS-1 on the proliferation and apoptosis of RA-induced PC12 cells . Methods The MTT assay, flow cytometry, TUNEL method were used in the research. Results An inhibition of proliferation was demonstrated in the RA-induuced PC12 cells expressing PS-1 mutant L286V. The proliferation of the cells was blocked in the G1→S phase. Whether with or without fetal bovine serum(FBS), PS-1 mutant L286V was able to accelerate the apoptosis of RA-induced PC12 cells. This action was enhanced markedly under the culture without FBS. Conclusions The RA-induced PC12 cells expressing PS-1 mutant L286V shows that the proliferation is inhibited and blocked in the G1→S phase. Whether with or without FBS, PS-1 mutant L286V can accelerate the apoptosis of RA-induced PC12 cells. This action is enhanced markedly under the culture without FBS.

Objective To investigate the cholinergic neuron properties and the changes of proliferation and apoptosis of PC 12 cell induced by different concentrations of all trans retinoic acid(RA). Methods PC 12 cell was induced by RA of concentrations of 1-50 ?mol/L. Changes of cell growth curve and cycle, apoptosis and changes of activities of acetylcholinesterase(AchE) and choline acyltransferase(ChAT) were detected by MTT assay, flow cytometry, TUNEL method, spectrophotometry and radiochemistry assay, respectively. Results ① RA significantly enhanced the activity of ChAT with concentrations between 1 to 20 ?mol/L. The activity of ChAT with concentration of 10 ?mol/L was higher than those with other concentrations. ② The apoptosis of RA induced PC 12 cells was markedly enhanced when concentration was higher than 10 ?mol/L. ③ A concentration dependent inhibition of proliferation was demonstrated in the RA induced PC 12 cells. ④ PC 12 cells proliferation was blocked in the G 1→S phase. Conclusion RA can be used to induce PC 12 cells to show the properties of cholinergic neuron with an optimal concentration of 10 ?mol/L.

Objective To observe the sensitivity ofastrocytoma to chemotherapeutic drugs, in order to explore the drug selection using chemosensitivity testing in vitro on astrocytoma. Comparing different WHO grade astrocytoma to chemotherapeutic drugs sensitivity differences,providing the basis data for clinical treatment. Methods The astrocytoma cells were seperated and cultured in vitro from 142 tumor tissues by glioma resection, including 55 cases of low-grade astrocytoma (WHO grade Ⅰ , Ⅱ) and 87 cases of high-grade astrocytoma (WHO grade Ⅲ, Ⅳ). The proliferation inhibition and chemosensitivity of tumor cells to 11 drugs were investigated by using MTT method. Results There were 54.93% (78/142) patients recommended chemotherapeutic drug in 142 cases. The percentage was 41.82%(23/55) in the low-grade astrocytoma, and 63.22% (55/87) in the high-grade astrocytoma respectively. Teniposide,carboplatin,vincristine as the sensitive drugs were similar in different WHO grade astrocytoma. Conclusions The chemosensitivity testing could be used to filtrate sensitive drugs to clinical chemotherapy. The action is more prominent in high-grade astrocytoma than in low-grade astrocytoma. There is no difference in the choice of sensible drugs among different WHO grade astrocytoma.

Objective To approach the feasibility of transfecting the DNA plasmid of encoding red fluorescent protein directly into the nucleus of rabbit primary bone marrow stromal cell with recently developed nucleofection technique. Methods Rabbit primary bone marrow stromal cells (BMSCs) were harvested by means of density gradient centrifugation following a thighbone puncture. The primary BMSCs were cultured and either transfected to pDsRed1-N1 by nucleofectorTM technique (as DsRed group) or left uninfected(as control group) in vitro. The cellular viability, adhesive rate, the growth curves and the efficiency of transfection of both DsRed and control groups were analyzed. Result DsRed were successfully expressed at 48h after nucleofection. Similar morphology evolvement, adhesive rates and growth curves were obtained from the two groups. The positive DsRed expression enhanced gradually alone with a prolonged culturing time, and reached its peak value at the 10th day after marked, with about 54.2% of DsRed-positive cells in the total BMSCs. The DsRed did not attenuate even until 1 month following the mark. Conclusion Neuclofection of pDsRed1-N1 showed no significant effect on the proliferation of rabbit BMSCs. DsRed worked efficiently for the purpose of stable gene marking of rabbit BMSCs, and nucleofection is an efficient method for transferring genes into primary rabbit BMSCs.

Purpose To study the effect of China cobra venom active factor(CCVAF) from China cobra venom on endothelial cells and its mechanism.Methods MTT experiment was adopted to evaluate the effect of CCVAF on bovine arteria pulmonalis vascular endothelial cells(BAVEC).The Eosin-Coomassie brillient blue and rhodamine-phalloidin method was used for actin cytoskeleton.Flow cytometry for [Ca~(2+)]_i and spectrophotometry were used for lactate dehydrogenase(LDH) and nitrogen oxide(NO) levels in cell culture supernatant.Results CCVAF(0.625-20 μg/mL) inhibited the proliferation of BAVEC in dose-dependent manner,and IC50 of CCVAF on BAVEC was 2.45 μg/mL. After CCVAF and BAVEC coincubation, it was showed that regression of intercellular conjunctions and disorder of F-actin distribution occurred. The content of [Ca~(2+)]_i, [LDH] and [NO] increased respectively.Conclusion CCVAF can inhibit BAVEC proliferation and it maybe associated with the change of cytoskeleton and increasing of [Ca~(2+)]_i,[LDH] aod [NO].

Objective To inquire into the curative effects of combination of laproscopic cholecystomy and laparoscopic transcystic duct common bile duct exploration on cholelithiasis, cholecystolithiasis and cho-ledocholith. Methods The clinical data of 19 cases were retrospectively analyzed from Sep. 2006 to Jan.2009. Results All the operations were performed successfully. The operative time was 60～120 min, the drainage time was 3～5 d, and the postoperative hospitalization time 4～7 days. All of the patients were fol-lowed up from 3 to 6 months. No complications occurred. Conclusion Laparnscopic transcystic biliary duct exploration is safe, feasible and worth generalizing.

OBJECTIVE:To observe the efficacy and safety of doxofylline combined with ceftazidime in the treatment of elder-ly patients with chronic emphysema. METHODS:Data of 78 elderly patients with ceftazidime was collected and divided into single drug group(31 cases)and combined medication group(47 cases)according to different treatment methods. All patients adopted flu-id infusion,oxygen inhalation and nutrition support. Based on it,single drug group received 0.05 g Doxofylline injection,once ev-ery 12 h. Combined medication group additionally received 1.0 g Ceftazidime for injection,once every 12 h. If the symptoms were out of control,20 mg prednisone was given,twice a day. The courses of 2 groups were 10 d. Total efficacy,pulmonary functions [maximum mid-expiratory flow(MMEF),1s forced expiratory volume percent predicted(FEV1%),peak expiratory flow(PEF)] before and after treatment,American Concise Health Survey Questionnaire(SF-36 scale)(QLS)and the incidence of adverse reac-tions in 2 groups were observed. RESULTS:The total effective rate in combined medication group was significantly higher than sin-gle drug group,the difference was statistically significant (P<0.05). Before treatment,there was no significant difference in MMEF,FEV1%,PEF and QLS scores between 2 groups(P>0.05). After treatment,MMEF,FEV1%,PEF and QLS scores in 2 groups were significantly higher than before,and combined medication group MMEF,FEV1%、PEF were higher than single drug group,the difference was statistically significant (P<0.05),but there was no significant difference QLS scores in 2 groups(P>0.05). And there was no significant difference in the incidence of adverse reactions in 2 groups (P>0.05). CONCLUSIONS:Based on conventional treatment,doxofylline combined with ceftazidime shows better efficacy than doxofylline alone in the treat-ment of elderly patients with chronic emphysema,it can improve pulmonary functions,with better safety.

Objective To investigate the anti-hepatocarcinoma(anti-HCC) function of HLA-A2-restricted point-mutated Survivin peptide induced CTLs.Methods The HLA-A2-restricted Survivin nonapeptides were evaluated using bioinformatics software.The binding affinity of Survivin peptide to HLA-A2 molecular was determined with flow cytometry analysis.After peptide-induced CTLs were generated in vitro,flow cytometry and ELISA were performed to detect the levels of IFN-γ,which were secreted by reactive CTLs.Peptide-induced CTLs were co-cultured with hepatoma cell lines HepG2 and BEL-7402.The rates of tumor cells lysis were assayed using CytoTox 96(R) and the morphological changes of tumor cells were observed with inverted phase contrast microscope.Results Point-mutated Survivin nonapeptide Sur79M2 (KMSSGCAFL) was filtered out,which was shown higher scores compared with the wild-type peptide Sur79.Consistent with the results of software analysis,Sur79M2 showed higher binding ability in T2 binding assays.At the same time,Sur79M2-induced CTLs could release a large number of IFN-γ after incubated with target cells rather than Sur79.When co-cultured with HCC cell lines HepG2 and BEL-7402,Sur79M2-induced CTLs effectively lysis HepG2 on HLA-A2-restricted manner without killing effect on BEL-7402 that do not express HLA-A2 molecules.Conclusion Sur79M2 could elicited specific cytotoxic T lymphocytes in vitro,which were able to specifically kill HCC cell lines on HLA-A2-restricted manner.

Objective To investigate the clinical features and microsurgical resection techniques of cerebellopontine angle cholesteatoma. Methods Clinical features, surgical approach and techniques, results and complications were analyzed in 30 cases of cerebellopontine angle cholesteatoma. Results Total resection in 26 cases, subtotal resection in 4 cases ,no operative mortality. Post-operative complications was aseptic encephalitis which was most common after treatment, and all recovered. Conclusion Cerebellopontine angle cholesteatoma was a third category of common tumors, CT, MRI examination can provide diagnosis. According to cholesteatoma involving the site ,selecting a good surgical approach and making good use of microsurgical technique could improve surgical total removal rate and reduce postoperative complications.

OBJECTIVE: To observe the biological characteristics of the human olfactory mucosa mesenchymal stem cells (hOM-MSCs). METHODS: The hOM-MSCs were isolated, cultured and identified in vitro. Scanning electron microscope and transmission electron microscope were used to observe the ultrastructure of hOMMSCs. Th e cells were induced towards adipocyte, osteocyte, neural stem cells, neural-like-cells in vitro. RESULTS: The hOM-MSCs were mainly in spindle shape, arranged with radial colony. The hOMMSCs expressed CD73 and CD90 but no CD34 and CD45. Th e short and thick microvilli processes were seen at the surface of hOM-MSCs by scanning electron microscope, and 2 different cellular morphology of hOM-MSCs were seen under transmission electron microscope. Moreover, the hOMMSCs could be differentiated into adipocyte, osteocyte, neural stem cells and neural cells. CONCLUSION The hOM-MSCs possess general biological characteristics of MSCs and display multiple differentiation functions. They can be served as ideal seed cells in tissue-engineering for injury repair.
Cell Differentiation ; Cells, Cultured ; Humans ; Mesenchymal Stem Cells ; cytology ; ultrastructure ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Olfactory Mucosa ; cytology