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

Endoscopes ; Endoscopy ; Epistaxis ; diagnosis ; etiology ; therapy ; Humans ; Nose

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.

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

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 motor nerve function status in rats with diabetes mellitus by motor unit number estimation (MUNE), and discuss it′s early diagnostic value in diabetic peripheral neuropathy (DPN). Methods Diabetic rat model (DM group) was induced by streptozotocin. The MUNE of gastrocnemius muscle and motor nerve conduction (MCV, CMAP) of the sciatic nerve were measured at the 4th, 8th and 12th week after onset of hyperglycemia in the DM group and the control group (normal SD rats). The ultrastructure of sciatic nerve was observed by electron microscope. Results At the 4th week, MUNE of gastrocnemius muscle was significantly decreased in DM group compared to that of the control group (275.88 ± 87.87 vs 369.71 ± 75.64,P<0.05). There were no significant differences in MCV and CMAP of sciatic nerve be?tween two groups. The electron microscopy observation showed that most nerve fibers were normal;a small amount of axonal atrophy, and myelin lamellar structure was separated in DM group. At the 8th week, compared with the control group, MUNE were reduced in gastrocnemius muscle in DM group (357.49±72.68 vs 221.26±92.41, P<0.01). There were no significant dif?ferences in MCV and CMAP of the sciatic nerve between DM group and control group. The electron microscope observation showed that part of nerve fibers were normal, the myelin focal plate layer was loose and separated, axonal atrophy, the axonal membrane and myelin sheath inner layer was separated with big gap. At the 12th week, MUNE of gastrocnemius muscle (127.87±19.80 vs 366.85±51.25), sciatic nerve MCV [(35.06±4.43) m/s vs (50.47±6.07) m/s] and CMAP [(2.91±1.37) mV vs (5.98±2.14) mV] were significantly decreased in DM group than those of control group (P<0.01). The electron microscopy observation showed severely damaged myelin flex and axonal squeeze. Conclusion MUNE is much earlier in detecting ear?ly motor nerve dysfunction in DM than conventional motor nerve conduction test.