D-D4FC (?-D-2′,3′-didehydro-2′,3′-dideoxy-5-fluorocytidine),a new anti-HIV drug,is on its PhaseⅡ clinical trials in America,France and Germany. Our lab has synthesized D-D4FC successfully using N-deoxyribosyltransferase from Lactobacillus helveticto catalyzing the ribose transfer from D4T (?-D-2′,3′-unsaturated thymidine) to 5-FC (5-fluorocytidine).The yield of D-D4FC reached 25%.We discovered the reaction could also be done by using intact cells.The yield could increase to 50% in 12.5 hours and more convenient to industrial continuous process.In this paper,the conditions including pH,buffer,substrates concentration,cells amount,reaction time and a possible catalytic mechanism were studied and discussed.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; adverse effects ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; Cisplatin ; administration & dosage ; adverse effects ; Deoxycytidine ; administration & dosage ; adverse effects ; analogs & derivatives ; Drug Chronotherapy ; Female ; Humans ; Leukopenia ; chemically induced ; Lung Neoplasms ; drug therapy ; pathology ; Male ; Middle Aged ; Nausea ; chemically induced ; Neoplasm Staging ; Neutropenia ; chemically induced ; Remission Induction ; Taxoids ; administration & dosage ; adverse effects

Adenoma, Pleomorphic ; surgery ; Adult ; Female ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; surgery

Coloboma ; Humans ; Infant, Newborn ; Male ; Optic Nerve ; abnormalities

OBJECTIVETo study the effects of low concentration dopamine(DA) on hydrogen peroxide-induced apoptosis in cultured rat cardiomyocytes as well as the possible molecular mechanisms.

METHODSCultured neonatal rat cardiomyocytes were randomly divided into the following groups: control group (control), hydrogen peroxide group (H2O2), pretreated with low concentration dopamine ( DA + H2O2), dopamine receptor l(DR1) antagonist group (DR1 + DA + H2O2), dopamine receptor 2(DR2) antagonist group (DR2 + DA + H2O2). The cell apoptosis was then assessed by MTT and flow cytometry. The cellular ultrastructure changes were observed by transmission electron micro- scope. The activity of lactate dehydrogenase(LDH )and superoxide dismutase (SOD) in cell medium was analyzed by colorimetry. The protein expressions of Cytochrone c, Caspase 3 and Caspase 9 were obtained by Western blot.

RESULTSCompared with hydrogen peroxide group, low concentration dopamine(10 µmol/L) decreased the apoptosis rate and the expression of protein of apoptosis related protein, enhanced SOD activity, decreased LDH activity. DR1 antagonist SCH-23390 treatment inhibited dopamine induced cardiac protective effect. DR2 antagonist haloperido treatment had no changes compared with dopamine group.

CONCLUSIONAbove findings indicate that low concentration dopanine inhibits apoptosis induced by hydrogen peroxide in neonatal rat cardiomyocytes, which is partly associated with the activation of DR1.

Animals ; Apoptosis ; Benzazepines ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cells, Cultured ; Dopamine ; pharmacology ; Hydrogen Peroxide ; L-Lactate Dehydrogenase ; metabolism ; Myocytes, Cardiac ; drug effects ; Rats ; Rats, Wistar ; Receptors, Dopamine D1 ; metabolism ; Superoxide Dismutase ; metabolism

AIM: To investigate the variation and significance of mRNA and protein concentration of myeloid cell leukemin-1 (Mcl-1) in apoptotic HepG2 cells induced by apoptin. METHODS: The apoptin expression vector pCDNA3.0-VP3 was transfected into HepG2 cells via liposome. Mcl-1 mRNA was analyzed by real-time quantitative reverse transcriptase-polymerase chain reaction. The protein of apoptin, Mcl-1 and cytochrome C were detected by Western blotting. RESULTS: The VP3 gene was transfected into HepG2 cells successfully and expressed steadily. Compared to blank control, Mcl-1 mRNA and protein levels of VP3 positive cells were decreased (mRNA: 0.09%±0.00% vs 0.41%±0.14%, P<0.05; protein: 0.43%±0.01% vs 0.90%±0.04%, P<0.01). Released cytochrome C from mitochondrion was increased (0.98%±0.02% vs 0.62%±0.03%, P<0.01). CONCLUSION: In the course of the apoptosis of HepG2 cells induced by apoptin, the amount of Mcl-1 mRNA and protein is decreased, and released cytochrome C from mitochondrion is increased. The apoptosis induced by apoptin may be correlated with the down-regulation of Mcl-1 mRNA and protein.

Objective Using mouse autoimmune premature ovary failure (POF) model to seek theoretical evidence for a possible clinical therapy of autoimmune POF with glucocorticoid (GC) or an-drogen. Methods After autoimmune POF was induced in 60 mice by Pzp3, the mice were randomly as-signed into 3 groups (n=20) : Two groups were treated with GC or androgen and the control group was treated with distilled water. We observed the changes in the sexual cycles of the mice, the serum level of AzpAb, infiltration of cells positively expressing CD45 in the ovary, and pathological alterations of the ovary. Results The sexual cycle of each therapy group was significantly different from that of the control group. The mean serum level of AzpAb of each therapy group was significantly lower than that of the con-trol group, and the mean serum level of AzpAb in the GC group was significantly higher than that of the androgen group. The percentage of growing follicles in the ovary of each therapy group was significantly higher than that of the control group. Ovaries infiltrated by cells positively expressing CD45 of each thera-py group were significantly fewer than those of the control group. Conclusion GC or androgen in mice with autoimmune POF could obviously ameliorate the pathogenetic conditions of the disorder, and both treatments have similar therapeutic efficacy.

Objective To explore the effect of endoscopic sinus surgery combined with postoperative nasal irrigation on the symptoms improvement and prognosis for patients with allergic fungal sinusitis.Methods The clinical data of 145 patients with allergic fungal sinusitis who had received treatment in our hospital from January 2010 and January 2014 were reviewed.According to different treatment methods,they were divided into two groups,65 cases in the control group was treated with conventional surgical treatment,80 cases in the obeservation group was treated with endoscopic sinus surgery combined with postoperative treatment.The therapeutic conditions of two groups were compared. Results The symptom scores of patients with polyps,edema,rhinorrhea in two groups were decreased compared with those before treatment, and the obeservation group decreased more significantly (P <0.05,P <0.01).After the operation,patients with nasal mucociliary transport rate of two groups were higher compared with postoperative 6 months and 9 months,and the obeservation group improved more significantly (P <0.05).In addition,two groups of postoperative SNOT-20 scores (physiological problems,limitations and affective outcomes)treatment were lower than those before,and the obeservation group decreased significantly than that of the control group (P <0.05,P <0.01).Con-clusion Endoscopic sinus surgery combined with postoperative nasal irrigation can effectively alleviate the symptoms and improve the prog-nosis in the treatment of allergic fungal sinusitis.

Objective To investigate the effect on apoptosis and extracellular signal regulated kinase (ERK) expression of cholangiocarcinoma RBE cell line induced by paclitaxel and the role of extracellular signal regulated kinase in paclitaxel-induced apoptosis. Methods By techniques of cell culture in vitro, annexin-V-FITC and propidium iodide(PI)double staining and flow-cytometry (FCM), RBE cell line was treated by paclitaxel or together with MEK inhibitor PD98059 for 24 h. Using Western blot test the expression of ERK1, ERK2, pERK after RBE cell line was treated by paclitaxel or together with PD98059. Results Paclitaxel induced RBE cell line to apoptosis, the ratio of apoptosis was 14.8%?2.7%(n=6), the control apoptosis was 3.4%?1.3%(n=6,P

Objective To observe the expression of p38MAPK activity during mitomycin-induced cell cycle arrest of cholangiocarcinoma. Using p38MAPK inhibitor SB203580 to analyze the effect on chemotherapy of cholangiocarcinoma. Methods The selected human cholangicarcinoma cell lines RBE was treated with mitomycin or together with SB203580, cell apoptosis was analysed by AnnexinV-FITC and propidium iodide(PI)double staining and flow cytometry (FCM), p38 and ATF-2 activity was detected by Western blot. Results Mitomycin induced apoptosis of RBE cell line, the ratio of apoptosis was (8.91?0.83)%( P