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

This study aims to establish a method to determine the serum acetaminophen concentration based on diazo reaction, and apply it in the gastric emptying evaluation. Theoretically, acetaminophen could take hydrolysis reaction in hydrochloric acid solution to produce p-aminophenol, which could then take diazo reaction resulting in a product with special absorption peak at 312 nm. Then the serum acetaminophen concentration and recovery rate were calculated according to the standard curve drawn with absorbance at 312 nm. ICR mice were given a dose of acetaminophen (500 mg x kg(-1)) by gavage and the serum acetaminophen was dynamically measured through the diazo reaction. Besides, ICR mice were subcutaneously injected with the long-acting GLP-1 analog GW002 before the gavage of acetaminophen, and serum acetaminophen concentration was measured as above to study how GW002 could influence the gastric emptying. The data showed acetaminophen ranging from 0 to 160 μg x mL(-1) could take diazo reaction with excellent linear relationship, and the regression equation was y = 0.0181 x +0.0104, R2 = 0.9997. The serum acetaminophen was also measured with good linear relationship (y = 0.0045 x + 0.0462, R = 0.9982) and the recovery rate was 97.4%-116.7%. The serum concentration of acetaminophen reached peak at about 0.5 h after gavage, and then gradually decreased. GW002 could significantly lower the serum acetaminophen concentration and make the area under the concentration-time curve (AUC) decrease by 28.4%. In conclusion, a method for the determination of serum acetaminophen based on the diazo reaction was established with good accuracy and could be used in the evaluation of gastric emptying.
Acetaminophen ; blood ; pharmacokinetics ; Aminophenols ; Animals ; Gastric Emptying ; Mice ; Mice, Inbred ICR

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 study the roles of the contact heat evoked potential (CHEP) in evaluating the small nerve fibers of cranial and spinal in elderly diabetic patients,and to analyze the feature of the small fibers in order to provide scientific foundations for early diagnosis of diabetic neuropathy (DN).Methods Totally 50 diabetic patients and 40 healthy subjects were included in this study.The diabetic patients were divided into the normal nerve conduction group (n=17) and the abnormal nerve conduction group (n=33).CHEP was determined and thermal stimuli was given to the area of eyelid,forearm and peroneal.Sensory and motor nerve conduction velocity in the upper and lower limbs were tested in all diabetic patients.Results Compared with the healthy group,the N wave latencies were prolonged and the N-P amplitudes were reduced in the area of eyelid,forearm and peroneal in diabetic patients (all P＜0.01).Compared with the healthy group,the logarithm of N-P amplitudes were reduced in areas of forearm and peroneal [(1.70±0.10) vs.(1.60±0.14),(1.65±0.078) vs.(1.54±0.15),both P＜0.05] and N wave latencies were prolonged in eyelid area [(343.1±18.2) ms vs.(385.4±26.5) ms,P＜0.05] in normal nerve conduction group.Compared to group with normal nerve conduction velocity,the logarithm of N-P amplitudes was reduced in peroneal area [(1.54±0.15) vs.(1.44±0.15),P＜0.05] in abnormal nerve conduction group.Conclusions CHEP can detect the early impairment of the small fibers in elderly diabetic patients.The early smallfiber impairments in diabetic neuropathy may be axon-oriented in spinal nerves and myelin-oriented in cranial nerves.

Objective To assess the function of motor nerve fiber in patients with diabetic peripheral neuropathy (DPN) by single-fiber conduction studies.Methods According to the diagnostic standard of DPN issued in Toronto meeting in 2009,on the basis of the result of peroneal nerve conventional conduction study,a total of 65 patients with DPN in the Department of Endocrinology and the Department of Neurology of Tianjin Third Central Hospital from October 2012 to October 2013 were enrolled into the study,from whom 33 had abnormal sensory conduction (sensory-diabetic peripheral neuropathy group,S-DPN group),32 had abnormal sensory motor conduction (sensory motor-diabetic peripheral neuropathy group,SM-DPN group).Single-fiber conduction velocity (SF-CV) and single-fiber distal motor latency (SF-DML)were detected in all subjects.The obtained results were compared with the data from 34 healthy volunteers (control group).The relationship of SF-CV,SF-DML and the duration of diabetes mellitus,fasting glucose,HbA1 c was also studied in DPN patients.Results The SF-CV ((43.1 ± 3.6) m/s) was decreased in S-DPN group compared with control group ((47.5 ± 3.3) m/s,t =5.077,P ＜ 0.01).There were no significant differences in SF-DML ((3.6 ± 0.7) ms),motor nerve conduction velocity (MCV (49.5 ± 2.6)m/s) and DML ((3.4 ± 0.6) ms) in S-DPN group compared with that of control group ((3.4 ± 0.5) ms,(50.9 ± 3.5) m/s,(3.2 ± 0.5) ms,respectively).SM-DPN group had lower SF-CV ((35.2 ± 3.6)m/s,t =9.119,14.219),MCV ((40.9 ± 3.2) m/s,t =11.131,13.025) and increased SF-DML ((4.5±0.7) ms,t=5.692,7.231),DML ((4.2 ±0.7) ms,t=5.561,6.975) compared with the other two groups (P ＜0.01).SF-CV in DPN patients was negatively related to the diabetic duration (r =-0.340,P =0.006),while SF-DML had no correlation with duration of DM,fasting blood glucose and HbAlc.Conclusions Detection of SF-CV is easy to find early motor nerve dysfunction in DPN patients.SF-CV is decreased with the increasing duration of diabetes.