Objective To study how polyclonal antibodies against ciliary neurotrophic factor (CNTF) reduce botulinum toxin-in- duced axonal sprouting and affect paralytic muscle.Design Experimental study.Participants 20 New Zealand rabbits.Methods In 10 rabbits randomly selected,left superior rectus were as control (Group 1),right superior rectus were injected with botulinum toxin A (BTXA) 2.5U (Group 3).At 14th day,bilateral superior recti were taken out under general anaesthesia.In the other 10 rabbits,left su- perior recti were injected equivalent physical saline (NS) as control (Group 2);right superior recti were injected with BTXA 2.5U,after 4 days,injected with 50?l polyclonal antibodies against CNTF at same site (Group 4).And at 14th day,bilateral superior recti were taken out for electron microscopy,dyed with acetylcholinesterase,argentums (Ag) to show nerve axonal sprout,and accounted and mea- sured with Leica microsystems.Main Outcome Measures The mean number of sprouts and the mean total length of sprouts,and the uhrastructure change of muscle by electron microscopy.Results In Group 1,the mean number of sprouts and the mean total length of sprouts were 5.75% and 10.53?m respectively;Group 2 were 6.11% and 11.16?m;Group 3 were 84.04% and 170.71?m;and Group 4 were 54.77% and 68.12?m.The differences were statistically significant (all P=0.000).Electron microscopy showed that after admin- istration BTXA alone (Group 3),muscle atrophied obviously,nerve myelin sheath increased,while the structure of nerve and muscle re- main invariable.The Group 4 showed that local myofilament disrupted and dissolved,degenerative myocytes necrotized and disintegrat- ed into fragments,which led to partial unreversible destroy.Conclusion Polyclonal anti-CNTF can reduce BTXA-induced axonal sprout- ing,lead to partial unreversible destroy of muscle,which may prolong the time of paralytic muscle resuming.It suggests that polyclonal anti-CNTF could prolong the duration of muscle paralyses induced by botulinum toxin.

Central Nervous System Neoplasms ; secondary ; Humans ; Neoplasm Metastasis ; Neuroblastoma ; pathology

China ; Conservation of Natural Resources ; Electricity

OBJECTIVE To investigate the effect of cadmium (Cd) exposure in vivo and in vitro on duodenal epithelial cells in mice and the mechanism. METHODS In vivo, C57BL/6 mice were ig administered with CdCI210 mg-kg" once per day for 30 d to establish a chronic cadmium poisoning model, or were ig administered with a single dose of CdCh 80 mg • kg-1 to establish an acute cadmium poisoning model before the survival status and survival rate of mice were observed. Duodenal epithelial cells of acute and chronic cadmium poisoning mice were isolated and cultured. The cells were identified as epithelial cells by E-cadherin immunofluorescence. Then, the content of cadmium ion in duodenal epithelial cells was detected by confocal laser microscopy. In vitro, duodenal epithelial cells of normal C57BL/6 mice were isolated and cultured, and incubated with CdCI2 2.5-100 pmol-L-1 for 24 h. CellTiter-Blue was used to detect cell viability. Subsequently, the duodenal epithelial cells of normal mice were incubated with CdCI215 Mmol • L-1 for 24 h, and cell cycle was detected by flow cytometry. Then, the duodenal epithelial cells of normal C57BL76 mice were incubated with CdCI230 Mmol-L'1 for 3-12 h and the expression of cleaved-caspase 3 was detected by Western blotting. RESULTS Compared with the control group, the activities of mice with chronic cadmium poisoning were all normal. The mice with acute cadmium poisoning showed depression, less food intake and death. At the 5th day of acute cadmium exposure, the survival rate of mice decreased to 40%. The content of cadmium ion in duodenal epithelial cells of acute and chronic cadmium poisoning mice increased significantly (F<0.01). Furthermore, CdCI2 inhibited the viability of duodenal epithelial cells cultured in vitro and the half inhibitory concentration (IC50) was (24.55±0.84) pmol-L-1. Compared with the control group, CdCI2 blocked the cell cycle of duodenal epithelial cells at Go/G, phase (P<0.05), and the expression of cleaved-caspase 3 in duodenal epithelial cells was significantly increased after CdCI2 treatment for 6, 9 and 12 h (P<0.05, P<0.01). CONCLUSION Cadmium that enters the body through the digestive tract can be absorbed by duodenal epithelial cells and cause damage to the cells. The mechanism of cadmium-induced damage may be related to cell cycle arrest and caspase 3-mediated apoptosis.

Objective To clone the alliinase gene from the garlic bulb and construct the eukaryote expression plasmid for expressing the recombinant alliinase in Pichia pastoris system and analyzing its bioactivity. Metheds The alliinase gene was cloned from the Zhejiang garlic bulb by RT-PCR and the eukaryote expression plasmid of alliinase was constructed with the pPIczαC vector. The recombinant plasmid was transformed into Pichia pastoris X-33 by eletroporation. The positive clones were screened and were induced by methanol. Supernatants after induction were analyzed by SDS-PAGE and Western blotting. The activities of the recombinant protein and the extracted alliinase were detected by the pyruvic acid method and compared by specific activity. The contents of the two kinds of alliinase were detected by Lowry method. Results The alliinase gene was successfully cloned from the garlic bulb, the length of alliinase gene was 1 500 bp, the molecule of the recombinant alliinase was about 5.5 × 104, existed in the supernatant of Pichia pastoris. The specific activity of the recombinant protein was (82.09 ± 3.89) U/mg and the nature alliinase was (176.49 ± 5.06) U/mg. Conlusion The alliinase gene is successfully expressed in Pichia pastoris system. The recombinant alliinase has the activity of enzyme, but is lower than that of the extracted alliinase.

Drugs, Chinese Herbal ; pharmacology ; Humans ; Kidney Failure, Chronic ; mortality ; physiopathology ; Renal Dialysis ; methods ; mortality ; Survival Rate

ObjectiveTo investigate the efficacy and toxicity of docetaxel combined with compound tegafur capsule(S-1)on anthracycline-refractory recurrent metastatic breast cancer (ARMBC).Methods Thirty-eight ARMBC patients were given intravenous 70 mg/m2 docetaxel at day 1,and oral 60 mg/m2 S-1twice every day at day 1 to 14.Every 3 weeks was one cycle and each patient received at least two cycles.ResultsAfter treatment,among these 38 patients,there was 2 complete response (CR) (5.3 ％),20 partial response (PR) (52.6 ％),10 stable disease (SD) (26.3 ％),and 6 progressive disease (PD) (15.8 ％).Overall objective response rate was 57.9 ％ (95 ％ confidence intervaal: 42.6 ％-74.2 ％) while clinical benefit response rate was 73.7 ％ (95％ confidence interval: 58.4 ％-89.1％).The median time to progression (TTP) was 7.8 months(95 ％ confidence interval:6.7-8.9 months),and median overall survival time(OS)was 15.7 months (95 ％ confidence interval: 12.9-18.8 months).The main toxic reaction was myelosuppression,and grade Ⅲ and Ⅳ adverse events including leucopenia occurred in 21.1％ of all cases.Most common grade Ⅰ and Ⅱ adverse events,such as hand-foot syndrome,nausea,vomiting,diarrhea,liver dysfunction,and oral mucositis,were tolerable.ConclusionGood clinical efficacy is achieved in the therapy of metastatic breast cancer with docetaxel and S-1 combination regimen and toxic reaction is tolerable.

Objective To investigate the effect of xuebijing injection applied after abdominal operation on the inflammatory reaction,infection rate,length of stay and hepatorenal function. Methods Sixty patients received abdominal operations were randomly divided into therapeutic group (30 cases) and control group (30 cases). The inflammatory reaction indexes (temperature,heart rate,leucocyte) and hepatorenal function indexes(aminopherase, total bilirubin, creatinine, blood urea nitrogen) were observed in both groups before operation and after operation 3,7 days respectively. The infection rate and length of stay were compared in two groups. Results Temperature and leucocyte post operative 3 days [temperature:(37.0 ± 0.2) ℃ vs. (37.9 ± 0.4) ℃ ;leucocyte: (8.8 ± 1.1 ) × 109/L vs. ( 10.3 ± 1.7) × 109/L, P＜ 0.05] and length of stay in therapeutic group was obviously better than that in control group[ (8.7 ± 1.9 ) d vs. ( 10.9±1.6) d, P ＜ 0.05 ]. The hepatorenal function indexes and infection rate in both groups had no significant difference(P ＞ 0.05). Conclusion Xuebijing injection could significantly palliate the inflammatory reaction after abdominal operation, shorten the length of stay and has no hepatorenal toxicity in short term.

Objective To set up a novel clinical method for storage of oocytes by slowly freezing and successful pregnancy by intracytoplasmic sperm injection (ICSI). Methods The frozen oocytes (n=33) in metaphase Ⅱ (28),which were normal in morphology,were denuded of their cumulus-corona complex.The protective freezing solution contained 1,2-propanediol (1.5 mol/L) and sucrose (0.35 mol/L) and the process was slow for freezing and rapid for thawing.The recipients of these oocytes received hormone replacement therapy (HRT). Results A high survival rate of oocytes (32/33) was obtained when the sucrose concentration was 0.35 mol/L.ICSI induced a higher fertility rate (23/28),a good embryo cleavage rate (21/23) and a satisfactory embryo morphology for thawed oocytes.Embryo transfers were performed in 6 cycles/6 cases. Four cases got clinical pregnancy by demonstration of four gestation sacs on B ultrasonic examination 7 weeks after ICSI,1 fetal was first trimester spontaneous abortion,another 3 survival fetal were ongoing pregnancy and the first case gestation week is 17 weeks. Conclusions This study is the first successful application of human oocyte cryopreservation in China and this method can increase the survival rate of freezing oocytes.It’s easy to carry out,low in cost and high in the recovery rate of oocytes after thawing.