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 discuss the effects of extract of Ginkgo BilobaLeaves(EGb) on expression of cytokine of renal interstitial fibrosis induced by transforming growth factor-?1 (TGF-?1) and extracellular matrix. Methods Cultured human kidney cells(HKC) were divided into three groups: control group,TGF-?1(8 ng/mL) group,and TGF-?1(8 ng/mL) added EGb(25,50,100,150 mg/L)group.After 72 h,expression of ?-SMA was detected by cell immunochemistry ABC,and collagen type I by Real-time PCR and Western blotting. Results Treated with TGF-?1(8 ng/mL) for 72 h,expression of ?-SMA and collagen type Ⅰ were up-regulated markedly compared with control group.Treated with EGb(25,50,100,150 mg/L)and TGF-?1(8 ng/mL)concomitantly for 72 h,expression of ?-SMA and collagen typeⅠ were down-regulated in dosage dependent manner compared with TGF-?1 group. Conclusion EGb can inhibit expression of ?-SMA and collagen type I in HKC induced by TGF-?1,and the possible mechanism might be related to the inhibition of EGb on renal tubular epithelial-myofibroblast transdifferentiation.

Objective To observe the expression and location of TF and TFPI in femoral artery atherosclerotic plaque.Methods We detected the expressions and locations of TF and TFPI in femoral artery atherosclerotic plaque by immunohistochemical and double-stain immunohistochemical method.We detected TF mRNA and TFPI mRNA expressions in atherosclerotic plaque by RT-PCR,with the normal umbilical artery as a control.ResultsThe normal umbilical artery contained little TF,TFPI and their mRNA in the adventitia.A great deal of TF,TFPI and their mRNA were found in the tunica intima of the femoral artery atherosclerotic plaque.Conclusion Expression of TF,TFPI and their mRNA of all types of cells and stroma in the proliferative tunica intima.

Objective To determine the purity,composition and antitumor activities of polysaccharide AHP-12 from abalone harslet.Methods Its purity was checked by HPLC with TSK-GEL G4000PWXL column and agarose gel electrophoresis;Molecular weight was determined by gel filtration chromatography;Sulfate content was identified by gelatine nephelometry and aminohexose content by chromatometry;Gas chromatograph was applied to determine monosaccharide composition;Antitumor activities were investigated by MTT method.Results AHP-12 from abalone harslet was a homogeneous polysaccharide both measured by molecular weight and electric property;The molecular weight was about 3?105;It was contained sulfate 13.07% and aminohexose 4.98%;AHP-12 was composed of rhamnose,fucose and galactose(the ratio in mole is 1∶2.2∶1.7);the activity determined by MTT method showed it could hamper the growth of HeLa cell.Conclusion AHP-12 was extracted and purified from abalone harslet.It was a homogeneous polysaccharide,which contained sulfate and aminopolysaccharide,and with weak antitumor activities in vitro.