Objective To investigate the in vitro effect of sirolimus on the apoptosis of a cutaneous squamous cell carcinoma cell line, Colo-16 cells. Methods Cultured Colo-16 cells were treated with different concentrations (50, 100, 150, 200 nmol/L) of sirolimus for various durations ( 12, 24, 48, 72 hours). Subse-quently, cell proliferation was detected by MTT assay, and cell apoptosis by Annexin V-FITC and PI double staining. Morphological changes of the cells were observed with Hoechst 33258 fluorescent staining. Total RNA was extracted from Colo-16 cells treated with sirolimus for 48 hours, and subjected to reverse tran-scription (RT)-PCR for the detection of mRNA expression of B cell lymphoma/leukmia-2 (Bcl-2) and Bcl-2-associated X Protein (Bax). Results Sirolimus inhibited the proliferation of Colo-16 cells in a time-and dose-dependent fashion. The early apoptosis rate was 7.26%±0.26%, 8.34% ±0.19%, 9.86%±0.14%, 11.92% ±0.15% in Colo-16 cells treated with sirolimus of 50, 100, 150, and 200 nmol/L, respectively, signifi-candy higher than that in untreated cells (1.53%±0.09%, P < 0.05); a positive correlation was observed between the apoptosis rate and concentrations of sirolimus (r = 0.955, P = 0.000). Typical morphological changes of apoptosis, such as chromatin condensation and margination as well as nuclear fragmentation were observed by fluorescence staining. After treatment with sirolimus for 48 hours, a significant decrease was observed in the mRNA expression of Bcl-2, while an increase in that of Bax was noticed. Conclusion Sirolimus could induce Colo-16 cells apoptosis in vitro, which may be associated with the modulation of expression of apoptosis-regnlating genes, such as Bcl-2 and Bax.

Objective:To study the biological characteristics and mineralization activity of strom cells (MSC) isolated from rat bone marrow.Methods: MSCs were isolated from rat bone marrow using gradient centrifuge (460 g,20 min) with 70% percoll and cultured with DMEM. The cells were then incubated with mineralization culture medium (MCM) in the presence of Dex (10～7 mol/L), β-GP (10 mmol/L) and AcA (50 g/L).Cell growth was studied with cell counting,alkaline phosphatase (ALP) expression with a kit and mineralization with a saturated solution of alizarin red S (pH 4.3) stainning.Results: The isolated cells from bone marrow of rat were epithelial-like. The doubling time (DT) of the cells in MCM or DMEM were 50.3 h and 66.1 h respectively. Mineralization was observed after the cells had been cultured in MCM for 28 days. bFGF and rhBMP-2 increased ALP expression of MSC in a dosage dependent manner. Conclusion:In vitro cultured MSCs may be induced to differentiate towards osteoblasts.

The effects of polytetrafluoroethylene (e PTFE) membrane on periodontal ligament cell (PDLC) growth and periodontal tissues and new cement regeneration were studied, and the prospects of clinical application of e PTFE membrane were discussed. The results showed that PDLC grew well on the e PTFE membrane and no toxicity or inhibition was found. Close adhesion of the cells to the membrane was observed by scanning electron microscope. Histological studies suggested that the peculiar porous structure of e PTFE membrane protects and promotes healing of alveolar bone defect when the membrane edge was sealed perfectly on the crown side,exhibiting inhibition of growth of epithelium towards tooth root. The membrane was beneficial to regeneration of new alveolar bone and cement and establishment of new adhesion,and was regarded as a good substitute of old materials.

To summarize the application of proteomics to modernization research of Chinese materia medica (CMM) and offer the reference for modernization of CMM and new medicine preparation exploitation. Based on the references and our project group studies in proteomic of drug biosynthesis pathway and metabolization;the main content,technology strategies and important application of the proteomics were introduced significantly;and the further development of protemics was analyzed. The proteomics will become one of absolutely necessary tackles in the modernization research of CMM.

砄bjective:To study the biological characteristics and mineralization activity of strom cells (MSC) isolated from rat bone marrow.Methods: MSCs were isolated from rat bone marrow using gradient centrifuge (460 g,20 min) with 70% percoll and cultured with DMEM. The cells were then incubated with mineralization culture medium (MCM) in the presence of Dex (10～7 mol/L), ? GP (10 mmol/L) and AcA (50 g/L).Cell growth was studied with cell counting,alkaline phosphatase (ALP) expression with a kit and mineralization with a saturated solution of alizarin red S (pH 4.3) stainning.Results: The isolated cells from bone marrow of rat were epithelial like. The doubling time (DT) of the cells in MCM or DMEM were 50.3 h and 66.1 h respectively. Mineralization was observed after the cells had been cultured in MCM for 28 days. bFGF and rhBMP 2 increased ALP expression of MSC in a dosage dependent manner. Conclusion: In vitro cultured MSCs may be induced to differentiate towards osteoblasts.

To observe repair of dental defect and its curative effect of the vertical root split of molar in the seniors, every affected section of 36 cases (40 teeth) of the senior's root split was extracted by hemisection, the other sections of them were repaired by either complete crown restoration (15 cases) or fixed bridge restoration (21 cases). The results showed that after 6～12 months, excellent curative effect was achieved in 32 cases, good curative effect in 3 cases, poor curative effect in 1 case. Significant difference was found between the two repair methods( P

Objective To study the effect of hemisection in the treatment of the molars. Methods 136 molars were selected and hemisection technique were used. If the pathological change was concentrated to the root, such as periodontics, apical inflammation, root fractures, the pathological root was extracted. If the pathologicalchange was concentrated to the divergence of the roots or the bottom of pulp chamber, hemisection was performed. then all the tooth was restoredwith fixed restoration according to the solid conditions. Results These were followed up and evaluated in three years 94. 85% of the all molars were treated satisfactory. Conclusions The hemisection techniqueoperation is simple, but the indications are extensive,and its effect in the treatment of the molars is regarded as a positive method

Background and purpose:Hypoxia induced the decreased radiosensitivity of tumor cells, which was the cause of tumor radioresistance and relapse and metastasis. During the course, HIF-1a played the most important role in the regulation of hypoxia. However, it’s still unknown about the effect of HIF-1a on the radiosensitivity of hypoxia tumor cells and the relationship with autophagy. This study was to inhibit HIF-1αexpression in hypoxic lung adenocarcinoma cell line A549 with RNA interference (RNAi), and explore its effect on hypoxic cell radiosensitivity and autophagy. Methods: Plasmids pHIF-1α-shRNA and Neg-shRNA (negative control) were constructed and transfected into hypoxic A549 cells, this positive clone was named A549/HIF-1α-shRNA. Clone formation array was applied to calculate the value of D0, SF2, SER. The expression of HIF-1α, LC3, c-parp was detected by Western blot. Results:The SF2 of hypoxic A549 cell was 0.62, which was higher than that of normoxic A549 cell, SER was 1.45. The level of LC3Ⅱincreased significantly and the level of c-parp decreased after the radiation of hypoxic A549 cell. The level of HIF-1a increased in hypoxic A549 cells. The expression of HIF-1αin hypoxic A549 cells was suppressed markedly after transfection of HIF-1α-shRNA;this clone was named A549/HIF-1α-shRNA. The SF2 and SER were significantly lower in A549/HIF-1α-shRNA cells, 0.45 and 0.72 respectively. Under the hypoxic condition and after the inhibition of HIF-1α, the expression of LC3Ⅱ decreased significantly and the expression of c-parp increased. Conclusion:We successfully established a cell model that HIF-1αexpression was suppressed almost completely by RNAi. The inhibition of HIF-1αby shRNA may raise the radiosensitivity and decrease the autophagy of hypoxic A549 cells in vitro.

ObjectiveTo explore the relationship of the fragmented QRS (fQRS) and the fQRS time limit with ventricular arrhythmia in old myocardial infarction (OMI) patients through contrasting the incidence of ventricular arrhythmia in OMI patients whether fQRS or not and ventricular arrhythmia in different fQRS time limit.MethodsAccording to the routine electrocardiogram,321 OMI patients were divided into group A (fQRS appearance,167 cases) and group B(fQRS non-appearance,154 cases).The lead with fQRS extense was ehosen and traced another 50 mm/s electrocardiogram,and 3 consecutive fQRS time limit were measured and them average was taken in group A.According to the fQRS time limit,the patients in group A were divided into 3 groups: group X( ≤0.100 s,96 cases),group Y (0.101-0.119 s,54 cases) and group Z( ≥0.120 s,17 cases).All the patients were continuously monitored with 24 hours dynamic electrocardiogram,and the incidence of ventricular arrhythmia was analyzed.ResultsThe incidence of ventricular arrhythmia in group A [ 78.4％ ( 131/167 ) ] was higher than that in group B [ 63.6％(98/154) ] (P＜ 0.01 ).The incidence of premature ventricular contraction(PVC) ＞ 720/24 hours in group A [ 28.7％(48/167 ) ] was higher than that in group B[ 17.5％(27/154) ] (P ＜ 0.05 ).The incidence of multifocal PVC,coupled PVC,nonsustained ventricular tachycardia and Lown 3-5 grades PVC was 16.2％ (27/167),33.5％ (56/167),12.0％ (20/167),34.1％ (57/167) in group A,7.8％ (12/154),21.4％ (33/154),4.5％(7/154),23.4％(36/154) in group B,there were significant differences between two groups (P＜ 0.05 ).The incidence of ventricular arrhythmia in group Z [ 100.0％( 17/17 ) ] was significantly higher than that in group Y [79.6％(43/54)] and group X [74.0％(71/96)](P＜ 0.05).The incidence of Lown 3-5 grades PVC in group Z[ 70.6％( 12/17 )] was significantly higher than that in group Y[ 42.6％(23/54)] and group X [ 22.9％(22/96) ],and the incidence of Lown 3-5 grades PVC in group Y was significantly higher than that in group X (P＜ 0.05).ConclusionsOMI patients with fQRS have higher incidence and severe degree in ventricular arrlhythmia than those without fQRS.With the fQRS time limit widened,PVC and Iown 3-5 grades PVC significantly increased.So fQRS is a new predicting index of OMI,and fQRS time limit has definite value in predicting the heart event for OMI patients.

Objective To observe the clinical efficacy of the photon therapeutic apparatus on type 2 diabetic patients with periph-eral neuropathy .Methods While giving conventional nerve nourishing ,improving microcirculation treatment and nursing ,adjuvant therapy with Carnation-11 photon therapeutic apparatus was applied to one lower limb only of the 20 diabetes mellitus patients with peripheral neuropathy and compare the curative effects by paired observation of the two lower limbs .Results The evaluation score of peripheral neuropathy dropped obviously with ankle reflex and thumb vibration sensation improved greatly after 10 days of treat-ment and remained an unchanged follow -up test 30 days later ,which means the clinical therapeutic effect of the treated limb was significantly superior to the comparison limb .The difference of the curative effects is statistically significant ( P < 0 .05 ) . Conclusion Conventional therapy accompanied by adjuvant treatment of photon therapy can relieve the clinical symptoms and phys-ical signs of diabetic patients with peripheral neuropathy ,which could improve curative effect on diabetic peripheral neuropathy .