Substitution of thymidine with 5-Bromo-deoxyuridine (BUdR) was performed in vivo in mice for the detection of gossypol effects on the chromosome aberration and sister chromatid exchange (SCE) in spermatogonia. The male Kun-ming mice received a daily dose of 4mg/kg of gossypol for two weeks, followed by a single injec- tion intraperitoneally of BUdR adsorbed on activated charcoal. Metaphases from spermatogonia were prepared for observation 54 hours after the injection.The results demonstrated that the mean frequencies of chromosome aberration and SCE in spermatogonia of gossypol treated mice did not differ significantly from that of the controls. Whereas, the incidences of SCE in mice had been exposed to known mutagen mitomycin C which used as positive controls were markedly increased. This data provide evidence of more sensitive indices for chromosome damage to support the notion that gossypol was not a mutagenic agent.The method of in vivo analysis of SCE in spermatogonia and its application and significance in the detection of genetic damage of contraceptive agents was discussed.

Conservative sperm acrosomal antigens that react with a monoclonal sperm antibody against human spermatozoa HS63 were purified by immunoaffinity chromatography from rabbit testes. The purity and the molecular weight of the purified antigens were determined by SDS acrylamide gel electrophoresis. There is only a band on the acrylamide gel. The molecular weight is more than 60000. It is found that the antigen is a glycoprotein with the carbohydrate moiety of 17%. Following successive immunization antisera of high titers were raised and shown to react specifically with antigens on sperm acrosome, but not with any somatic cells as judged. By using mouse in vitro fertilization experiments and sperm penetration assay with zona-free hamster ova, the isoimmune sera from rabbit exhibited high degrees of fertilization inhibition as compared to the control sera. The results suggest that sperm-specific antigen reactive to HS63 may be a good candidates for the development of immunocontraceptive vaccines in humans.

Objective To study a new operative procedure for hydrocele in children under mini-lapascope.Methods 103 children with hydrocele aged from 1 to 9 years old were performed by suture around internal rings under mini-laparoscope from November 2000 to March 2002. Results The operative time was (5～8) minutes and hospital stay (4～5) days. The incision didn't need suture and there was no obvious scar after operation. All cases were followed up at 1st month, 6th month and 1st year postoperatively. All cases recovered except for 2 cases recurred at 1st month postoperatively. Conclusions Suture around internal rings under mini-laparoscope is a minimally invasive and simple method for the treatment of hydrocele in children. It can find and deal with hidden patent internal rings.

Objective To explore the clinical effects of laparoscopic hernioplasty in patient over 70 years old. Methods We utilized the modified laparoscopic intraperitoneal onlay mesh (IPOM) to treat the inguinal hernia in 69 elderly patients. Results The laparoscopic operations were completed in all the 69 patients. The operative time was 20～30 min (mean, 25 min). No operative complications took place and the intraoperative blood loss was hardly seen. The duration of hospitalization was 5～7 days. The patients recovered smoothly, without wound infection or scrotum hematoma. Follow-up observations for 6～24 months in the 69 patients found no recurrence. Conclusions Modified laparoscopic IPOM treating inguinal hernia in elderly patients is feasible, on the premise of proper perioperative management.

Objective To explore the clinical value of laparoscopy combined with radionuclide imaging in the diagnosis and treatment of pediatric gastrointestinal hemorrhage, especially for Meckel’s diverticulum and double intestine. Methods ~ 99m Tc~-pertechnetate abdominal scintigraphy was performed in 22 children with a history of recurrent hemafecia or melena in this hospital from December 1998 to December 2005. All the children were given a laparoscopic exploration. Results Among the 22 patients, scintigraphy showed positive findings in 18 patients and negative, 4 patients. The positive patients were all confirmatively diagnosed by laparoscopic surgery and pathological examinations, including Meckel’s diverticulum in 14 patients and duplications of alimentary tract in 4 patients. In the other 4 patients with negative results, no organic pathologic changes was identified by laparoscopy in 3 patients and Meckel’s diverticulum was found after laparoscopic exploration in 1 patient. Conclusions Radionuclide imaging is an important method in the diagnosis of gastrointestinal tract hemorrhage in children and provides scientific basis for surgical intervention. The combination of laparoscopy and radionuclide imaging not only has values in the diagnosis and treatment of pediatric gastrointestinal hemorrhage, but also minimizes the blindness and trauma of exploratory laparotomy.

We describe a modified technique for the in vitro study of human spermatozoal penetration into hamster oocytes which is easier to master and more simple to perform and needs no complicated facilities.It comprises the following aspects:1 The hamster oviduct is stained with 1％ neutral red agar slics, so that the opening of the oviduct is easy to find and easy to be inserted with needle for flushing eggs.2. A special small tube,3 cm long with a diameter of 0.5 cm is used for fertilization instead of plastic dish with paraffin oil covering the media.3. Ordinary incubator is used rather than CO_2 incubator. With these procedures, the sperm penetration rates we got were 75％, 25％, 40％, 80％ and 46％ respectively in 5 experiments. It could meet the needs to evaluate the fertilizing capacity of human spermatozoa.

Objective To investigate the methods and results of reverse island flap of the adjacent digit pedicled with the Y-V vascular of digital artery by anastomosis of superficial veins for repairing soft tissue defects of the fingers.Methods From March 2009 to June 2011,twenty cases with soft tissue defect distal to the proximal interphalangeal join of fingers were treated by reverse island flap of the adjacent digit pedicled with the Y-V vascular of digital artery by anastomosis of superficial veins.There were 12 cases of the index finger,eight of middle finger,the largest area of the flaps was 4.5 cm × 3.5 cm,and the smallest area was 3.5 cm × 2.5 cm,an average of the pedical length was 4.0 cm.All cases anastomosis one superficial vein,fourteen cases suture dorsal digital nerve,and the donor area covered with full-thickness skin graft.Results All flaps survived.Postoperative follow-up time ranged from 8 to 16 months,the appearance and texture of the flaps were excellent,the flaps with suture nerves,the two-point discrimination was 7 mm to 9 mm,the other flaps that the nerves were disconnected.The sensation of the flaps recovered to S2-S3,no morbidity of the donor fingers occurred.Conclusion Reverse island flap of the adjacent digit pedicled with the Y-V vascular of digital artery by anastomosis of superficial veins can form a longer vascular pedicle,to repair the soft tissue defect distal to the proximal interphalangeal joint,through anastomoses superficial venous can reduce the flap venous pressure obviously,improve the survival quality of the flap,the effect is satisfacted.

Background Oxidative stress is a pathophysiological process of retina,so it is very important to explore a protective way against retinal oxidative stress.Studies determined that extract of ginkgo biloba (EGb) has antioxidant,anti-apoptosis,anti-thrombosis and anti-inflammatory effects,however,the effect of EGb on human Müller cells in oxidative stress is still below understood.Objective This study was to investigate the protection of EGb against oxidative stress of human retinal Müller cells induced by As2O3 in vitro.Methods Human retinal Müllercell line was cultured in DMEM/F12 with 10％ fetal bovine serum (FBS),2 μmol/L glutamine and 1％ antibiotics.As2O3 solution at the final concentration 5 mg/L was added in the medium for 24 hours to establish oxidative models,and then the EGb with the final concentrations of 5,10 and 20 mg/L was used to cell models for 24 hours,respectively.Cell viability was detected by MTT assay,and reactive oxygen species (ROS) levels in the cells were detected with CM-H2DCFDA fluorescent probe.The relative expression levels of caspase-3 mRNA in cytoplasm and cell nuclei were assayed by reverse-transcription PCR (RT-PCR),and the expressions of Nrf2 protein were quantitatively detected by Western blot.Results Müller cells adhered well 24 hours after cultured.At 6-7 days after culture,Müller cell body was large with abundant cytoplasm and mosaic-like arrangement.However,floating cells were seen after As2 O3 treatment.Cell viability (absorbance) was significantly different among the normal culture group,As2 O3-treated group,As2 O3 + 5 mg/L EGb group,As2 O3 + 10 mg/L EGb group and As2 O3 + 20 mg/L EGb group,with the strongest viability in the normal culture group and the weakest viability in the As2 O3-treated groups (F=163.57,P =0.00).The fluorescence intensity of ROS was the weakest in the normal culture group and the strongest in the As2 O3-treated group and was gradually weakened with the increase of EGb doses,showing a remarkable difference among the groups (F =4 013.61,P =0.00).The relative expression level of caspase-3 mRNA in the cells was gradually reduced with the increase of EGb doses,with a statistically significant difference among the groups (F =2 199.72,P =0.00).In addition,no considerable difference was seen in the expression level of Nrf2 protein (grey scale) in cytoplasm among the groups (F=15.42,P=0.40);while in the nuclei,the expression levels of Nrf2 protein were 100.01 ±0.04,46.59±0.63,54.51 ±0.62,59.93 ±0.17 and 67.60±0.24 in the normal culture group,As2 O3-treated group and As2O3+5 mg/L EGb group,As2O3+10 mg/L EGb group,As2O3+20 mg/L EGb group respectively,with a significant difference among them (F=7 271.72,P=0.00).Conclusions EGb can protect human retinal Müller cells against As2O3-induced damage in a dose-dependant manner by antioxidant and antiapoptotic effects in vitro,and the activities occur primarily in cell nucleus.

Objective To investigate the method and result of repairing multi-fingers soft tissue defects using the dorsal metacarpal flaps with cutaneous branches as pedicle.Methods From February,2010 to January,2013,9 patients with multi-fingers tissue defects were treated with the 2nd,3rd,4th dorsal metacarpal flaps with cutaneous branches as pedicles.The area of flaps ranged from 1.2 cm × 2.5 cm to 2.5 cm × 5.0 cm.The donor sites were sutured with full thick skin graft.Results All flaps survived.After a followed-up of 8 months to 24 months(average 12 months),the texture and shape of the flaps were good and non-bloated.The flap sensibility as sessment were S3-S3+.The two-point discrimination testing were 10 to 13 mm (average 11.6 mm).The TAM score of range of motion was 60％ to 75％ of the healthy side.The skin graft of donor site were soft.Conclusion Procedure of dorsal metacarpal flaps with cutaneous branches as pedicles easy is a good method to repaire the soft tissue defects of muhi-fingers.

Background Oxidative stress is a main cause of age-related macular degeneration (AMD).Lutein has a preventive role for AMD, but its antioxidant mechanism remains unclear.Objective Present study was to investigate the effect of lutein on oxidative stress of Müller cells and its signaling pathway.Methods Human Müller cells (human Müller cell strain) were cultured, and the cells at logarithmimic growth phase were incubated in 96 well plate overnightly.Oxidative stress cell models were established by adding 160 μmol/L H2O2, a median lethal dose for Müller cells.The models were divided into the model control group and 12.5,25.0,50.0 mg/L lutein groups,and the different concentrations of lutein were used to culture the cells for 24 hours, respectively.The routine cultured cells served as the blank control group.Growth of the cells was assayed by MTT method (absorbancy);the reactive oxygen species (ROS) content in the cells was assayed by flow cytometry;the mRNA and protein levels of nuclear factor-E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) in the cells were detected by quantitative real-time PCR and Western blot, respectively.Results The inhibitory effects on the cells were gradually enhanced with the increase of H2O2 concentrations,showing a significant difference among the groups (F =43.890,P＜0.01).A significant difference was found in apoptotic rate of the cells among the blank control group,model control group and 12.5,25.0,50.0 mg/L lutein groups (F =346.770, P =0.000) , and the apoptosis rate was significant elevated with the increase of lutein dose (all at P＜0.05).The ROS contents in the cells were 1.92±0.18,64.89±2.86,52.70±2.80,32.61 ±4.20 and 5.68 ± 1.35 in the blank control group, model control group and 12.5,25.0,50.0 mg/L group, respectively, with significant difference among the groups (F =324.900, P =0.000), and the ROS content was gradually reduced as the increase of lutein dose (all at P＜0.05).The relative mRNA and protein expressions of Nrf2 and HO-1 were remarkedly higher in the 12.5,25.0,50.0 mg/L lutein groups than those in the model control group (F =236.960,242.620,186.830,263.120, all at P =0.000) , and no significant difference was seen in the relative expression level of nuclear Nrf2 protein among the groups (F =1.790, P =0.210).Conclusions Lutein can induce the expression of antioxidant enzymes by inducing the expression of nuclear translocation of Nrf2 and consequently inhibit the oxidative stress status.