Objective To study the apoptosis-inducing effect of histamin-indomethacin（his-IN）on Lewis lung cancer cell in mice.Methods The suspension of tumor cells from Lewis lung cancer mice was inoculated into the left axilla of Kunming mice.When the tumor grew to 1 cm in diameter,the mice were divided into 5 groups and received corresponding treatment：3.0 mg/kg indomethacin（IN）group,3.5 mg/kg IN group,3.0 mg/kg his-IN group,3.5 mg/kg his-IN group,and saline control group.Ten days later,the therapeutic effects were evaluated by the tumor inhibitory rate,the apoptotic peak and rate of tumor cells were measured by flow-cytometry（FCM）,and tumor necrosis factor α（TNF-α）level was estimated by radio-immunity method.Results TNF-α level,the tumor inhibitory rate and the apoptosis rate of his-IN group and IN group were significantly higher than those of the control group,and those of 3.5 mg/kg his-IN group was the highest.In the groups of his-IN or IN treatment,an apoptotic peak was found before G0/G1 phase,and the peak in his-IN group appeared earlier than IN group.Conclusion IN and his-IN could resist the growth of the tumor and promote cell apoptosis of Lewis lung cancer in mice.

OBJECTIVE:To optimize the extract technology of the total Favonoids from the fruits of Polygonum viviparum L.METHODS: The content of Flavonoids was computed with rutin as standard substance, and the influence of different factors and levels on the extract technology were investigated by means of simple factor experiment and orthogonal experiment. RESULTS: The optimum extraction technology was as follows: with water-bath temperature at 80℃, the ratio of liquid to solid material at 8∶1 and the concentration of ethanol at 50%, reflux extracting total Favonoids twice(1 h at each time). Under this condition, the content of total Flavonoids was 9.6%. CONCLUSION: The extraction method is appropriate, simple and feasible, and it provided theoretic basis for the production of this product.

Objective To study the chemical constituents of Clematis manshurica. MethodsThe extract from the roots and rhizomes of C. manshurica was subjected to chromatography on silica gel and Sephadex LH-20 column. The compounds obtained were identified on the basis of their physicochemical and spectroscopic evidences. ResultsEleven compounds were isolated and their structures were elucidated as squalene (Ⅰ), friedelin (Ⅱ), hexacosoic acid (Ⅲ), ?-sitosterol (Ⅳ), stigmasterol (Ⅴ), oleanolic acid (Ⅵ), ?-daucosterol (Ⅶ), 5-hydroxymethyl-2-furaldehyde (Ⅷ), methyl 3, 4-dihydroxy-phenyl lactate (Ⅸ), 5R-dihydro-5-hydroxymethyl-2(3H)-furanone (Ⅹ), 5R-5-hydroxymethyl-2(5H)-furanone (Ⅺ). ConclusionAll the compounds except for ?-sitosterol are isolated from the plant for the first time.

OBJECTIVE To analysis the effect of air disinfection by using nanometer light catalysis decontamination machine in operating room. METHODS By compare the effectiveness of air disinfection both by using nanometer light catalysis decontamination machine and ultraviolet rays light. RESULTS The result of tests is 0 CFU/m~2 by nanometer light catalysis decontamination machine and 33.3 CFU/m~2 by ultraviolet rays light in unmanned environment;By different groups: F=220.423,P=0.000,P

Objective:To establish an HPLC method for the determination of levofloxacin in levofloxacin hydrochloride ophthalmic in situ gel. Methods:The chromatographic column was Agilent Zorbax C18(150 mm ×4.6 mm,5μm). The mobile phase consisted of hexane sulfonic acid sodium solution-methanol(72∶28). The flow rate was 1. 0 ml·min-1. The detection wavelength was 293 nm. The chromatographic column temperature was 40℃. The injection volume was 10 μl. Results:The calibration curve was linear within the range of 0. 040 3-0. 403 0 mg·ml-1(r=1. 000 0) for levofloxacin. The average recovery was 99. 8% (RSD=1. 08%,n=9). Conclusion:The method is accurate, simple and rapid, and suitable for the content determination of levofloxacin in levofloxacin hydro-chloride ophthalmic in situ gel.

Objective To establish luciferase-labeled mesenchymal stem cells in vitro.Methods From May,2013 to January,2014,recombinant lentiviral vectors containing luciferase gene was transfected in to mesenchymal stem cell line C3H10T1/2.Puromycin was used to filter the cells.Western-blot and immunofluorescenceimaging were used to detect the transfection efficiency.Results Western blotting analysis showed that the size of this protein in cells expressing about 60KD.Showed that infected C3H10T1/2 cells expressing luciferase protein.I mmunofluorescence,compared with the uninfected control group,virus-infected cells group of about 100％ of the cells were positive for red fluorescence.Show that had built a good stable expression of luciferase C3H10T1/2 cells.Conclusion A monoclonal cell line stably and highly expressing luciferase is obtained with luciferase-labeled mesen chymal stem cells.

Objective To investigate mutations of CSMD3 gene in a pedigree of familial cortical myoclonic tremor with epilepsy (FCMTE).Methods Peripheral blood (5 ml) was obtained from FCMTE patients (7 cases),suspected cases,and control individuals.Polymerase chain reaction (PCR) and purification of PCR products for sequencing were used to detect the existence of mutations in 73 exons of gene CSMD3.The resulting products were subjected to agarose gel electrophoresis and gel-imaging system.The PCR amplification products were sequenced.Results The sequencing results of 73 exons were compared with CSMD3gDNA sequence in human GenBank.We neither found any DNA sequence variation nor disease-related mutations.Conclusions The family does not have a mutation in the CSMD3 gene.We need to further find the disease genes and the mutations in this family.

BACKGROUND: Prostate cancer stem cell is an important reason for the invasion and recurrence of prostatic carcinoma. However, separation efficiency of prostate cancer stem cells is very low.OBJECTIVE: To explore the efficient method for isolating and identifying the prostate cancer stem cells from human prostatic carcinoma cell lines PC-3 and LNCap. METHODS: Prostate cancer cell lines PC-3 and LNCap were cultured in serum free medium (SFM) and serum supplemented medium (SSM), respectively. The percentage of prostate cancer stem cells from different medium was detected by flow cytometry through markers CD133 and CD44, and the properties of prostate cancer stem cells were preliminarily identified using inducing differentiation experiments.RESULTS AND CONCLUSION: PC-3 and LNCap formed sphere cells in SFM, which can be induced into adherent cells after culture in SSM. Higher percentage of CD44+/CD133+cells was obtained from LNCap cells (1.71%; 0.73%) than PC-3 cells (0.59%; 0.32%) in both SFM and SSM. The number of CD44+/CD133+ LNCap cells was more than PC-3 using both methods (P < 0.05), but the efficiency of SFM and SSM had no statistical significance (P > 0.05). However, the culture cycle was longer and number of obtained cells was less by SFM culture, directly influencing functional determination of prostate cancer stem cells. Compared with suspension culture method with SFM, SSM is more convenient and effective in isolating prostate cancer stem cells from LNCap cells.

Objective To establish cell model of Parkinson's disease(PD)and to approach the toxic effect of rotenone on dopaminergic neurons and its mechanism.Methods PC12 cells were differentiated into dopaminergic neurons and treated with various concentrations of rotenone.The morphological changes of PC12 cells were observed after treated with rotenone(0,10,25,50,75,and 100 nmol?L-1)for 24,48 and 72 h,the cell viability was measured by MTT assay.Immunohistochemistry and immunofluorescence were used to observe the accumulation of ?-synuclein in cytoplasm.AO/EB double staining was also adopted to test apoptosis.Results After differentiation PC12 cells shaped irregularly with big and long ecptomas and multiple cell conjunctions.After the treatment of rotenone cell ecptomas vanished gradually and cell bodies became smaller and smoother.The cell viability began to decline significantly at a concentration of 50 nmol?L-1 for 24 h(P

Objective To evaluate the effect of salicylic acid on skin barrier function and the efficacy of salicylic acid combined with flumetasone ointment for the treatment of atopic dermatitis (AD).Methods Sixtyfour patients with AD (including 31 males and 33 females) aged 18 to 58 years were recruited into the present study.Four lesional areas of similar size and severity were selected at the similar body sites of both sides of each patient,and randomly classified into four groups to be topically treated with compound flumetasone ointment (containing 0.02％ flumetasone and 3％ salicylic acid,compound flumetasone group),flumetasone 0.02％ ointment (flumetasone group),salicylic acid 3％ ointment (salicylic acid group) and vehicle (control group),respectively;two normal skin areas were chosen from apparently normal skin on the similar body sites of both sides of each patient and topically treated with salicylic acid 3％ ointment (salicylic acid group) and vehicle (control group) respectively.All of these preparations were applied twice a day for 3 weeks.Transepidermal water loss (TEWL) was measured by a Tewameter MPA580 (Courage & Khazaka,Germany) at the baseline as well as on week 1,2 and 3 after initiation of treatment.Symptom and sign scores were evaluated before and after the treatment.Meanwhile,two normal skin areas were selected on bilateral forearm of 30 healthy controls and treated with 3％ salicylic acid ointment (salicylic acid group) and vehicle (control group) respectively twice a day for 3 weeks,and TEWL was measured before treatment as well as on week 1 and 3 after initiation of treatment.Results In the healthy controls,TEWL value showed no significant difference between the salicylic acid group and control group at any of these time points.As far as the lesional skin was concerned,no statistical difference was observed in TEWL value at the baseline between the four groups ((34.26 ± 20.82) vs.(33.02 ±16.71) vs.(34.16 ± 18.03) vs.(33.81 ± 17.11) g· m-2· h-1,P ＞ 0.05),but significant difference was noted after treatment (repeated measurement data analysis of variance,F =39.57,P ＜0.01),with the TEWL value being (22.38 ± 16.16),(17.04 ± 12.74),and (15.34 ± 13.13) g·m-2·h-1 respectively in the compound flumetasone group on week 1,2 and 3,(24.63 ± 17.08),(20.37 ± 9.53),(19.06 ± 9.17) g·m-2·h-1 respectively in the flumetasone group,(26.49 ± 8.59),(21.91 ± 8.46),(21.20 ± 9.38) g·m-2·h-1 respectively in the salicylic acid group,and (29.80 ± 12.48),(26.16 ± 8.31),(25.52 ± 6.05) g·m-2·h-1 respectively in the control group.In detail,the decrease in TEWL value was stronger in the compound flumetasone group than in the flumetasone group on week 1,2,and 3 (all P ＜0.05),in the salicylic acid group than in the control group (P ＜0.05 or 0.01),but similar between the flumetasone group and salicylic acid group.In non-lesional skin,the salicylic acid group showed a more intense decrease in TEWL value compared with the control group on week 2 and 3 (both P ＜0.05).Both the cure rate and response rate were significantly higher in the compound flumetasone group than in the flumetasone group (53.1％ vs.34.4％,x2 =4.57,P＜0.05;83.1％ vs.64.1％,x2 =6.90,P＜0.01).Conclusions The salicylic acid 3％ ointment shows a reparative effect on skin barrier in patients with AD,and the compound flumetasone ointment is superior to the flumetasone ointment in the treatment of AD.