Objective To establish a method for ex vivo cord blood stem cell expansion,increase the number of the stem/progenitor,especially megakaryocyte progenitors so as to decrease the thrombocytopenia period after CBT.Methods Mononuclear cells (MNC) from 20 cord blood samples were cultured in presence of different cytokines:group 1:IL 3,IL 6,group 2:IL 3,IL 6,TPO,group 3:IL 3,IL 6,TPO and cord blood plasma.The final concentration of the cytokines was 50ng/ml.The control group had all medium except the cytokines and cord blood plasma.The cells were cultured for 21 days.At every 7 days,tests were done for nucleate cells number,CFU GM,CFU Mk,and CFU GEMM.Results At different times and in different groups,there were different changes.For NC,maximal increase were obtained in group 3 on day 21,which was 239.65?153.48.For CFU GM,the biggest expansion occurred on day 21 in group 3,the number being 184.17?140.40.For CFU Mk,the biggest expansion was obtained on day 14 in group 3,with the number of 135.34?105.20. The immature progenitor CFU GEMM achieved the max increase in group 3 on day 14,the number was 49.39?36.68. Conclusion The expansion of CFU Mk progenitor cells can be obtained in the presence of IL 3,IL 6,TPO.This combination also enhances the expansion of CFU GM and CFU GEMM.Addition of cord blood plasma can promote the expansion of mature and immature progenitor cells.

Attention is being paid to the treatment of hyperopia(presbyopia) with laser thermokeratoplasty.This article mainly presents the theory of laser thermokeratoplasty(LTK),equipment configuration and some key techniques in laser delivery system.

Objective To discuss the etiology and treatment of extrahepatic biliary stenosis due to iatrogenic bile duct injuries. Methods A total of 12 cases of extrahepatic biliary stenosis after iatrogenic bile duct injuries(including 5 cases of open cholecystectomy and 7 cases of laparoscopic cholecystectomy) from January 1998 to January 2005 in this hospital was reviewed.After the establishment of a percutaneous transhepatic access,choledochoscopic stone removal,balloon dilatation,and drainage stent placement were performed for treating biliary stenosis.Results A F_(20) Gruntzig balloon catheter was employed for bile duct dilatation.The plastic drainage stent at 6~8 mm in diameter was successfully placed after 2 times of dilatation in 8 cases and after 3 times of dilatation in 4 cases,for 6~12 months of indwelling.Follow-up observations in the 12 cases for 2~3 years (mean,2.6 years) found no abdominal pain,fever,or jaundice.B-ultrasonography and MRCP findings showed no biliary stenosis and recurrent stones.Conclusions Choledochoscopic stone removal,balloon dilatation,and drainage stent placement are minimally invasive,safe,and effective in the treatment of extrahepatic biliary stenosis.

Objective To investigate methods and effects of percutaneous choledochoscopy via cholangio-jejunal anastomotic stoma in the treatment of recurrent intrahepatic biliary duct stones. Methods Eight cases of recurrent intrahepatic biliary duct stones after a Roux-en-Y cholangiojejunostomy from January 2000 to February 2005 were treated with percutaneous choledochoscopy. An enterostomy was made at the afferent jejunal loop. Then a choledochoscope was introduced into the intrahepatic biliary ducts by way of the cholangio-jejunal anastomotic stoma for stone removal and intrahepatic duct or anastomotic stoma dilatation. Results The intrahepatic stones were thoroughly removed in all the 8 cases. The biliary duct stenosis in 4 cases and the anastomotic stoma stenosis in 2 cases were relieved. Follow-up checkups in the 8 cases for 1~3 years revealed no recurrence of intrahepatic stones or stenosis. Conclusions Percutaneous choledochoscopy via cholangio-jejunal anastomotic stoma is minimally-invasive, safe, feasible, and effective in the treatment of recurrent intrahepatic biliary duct stones.

Objective: To investigate the characteristics of volunteer blood donors with regular apheresis platelets in Zhoushan City, Zhejiang Province, so as to provide insights into recruitment of regular blood donors. Methods: Gender, age, occupation, educational level, and volume and times of blood donation were collected from volunteer blood donors with apheresis platelets in Zhoushan Central Blood Station during the period from 2019 to 2021 through the Zhejiang Provincial Blood Management Information System BIS version 3.0, and the willingness of blood donation, routes for acquiring blood donation knowledge and cause of blood donation were collected from volunteer blood donors with regular apheresis platelets using questionnaire surveys. The factors affecting regular apheresis platelets were identified among volunteer blood donors using a multivariable logistic regression model. Results: Totally 236 volunteer blood donors participated in regular apheresis platelets in Zhoushan City from 2019 to 2021, accounting for 51.08% of all volunteer blood donors with regular apheresis platelets, and there were 1 488 blood donations, accounting for 71.23% of all volunteer blood donations with apheresis platelets. A total of 2 086.00 U platelets were donated, accounting for 71.82% of all volumes of volunteer blood donations with apheresis platelets. The volunteer blood donors included 160 men (67.80%) and 76 women (32.20%) and had an age of 35 (17) years. There were 63 employees (26.69%) and 111 with a bachelor degree and above (47.03%). Multivariable logistic regression analysis showed a high proportion of regular blood donations among participants at ages of 31 to 60 years (OR:1.767-3.257, 95%CI: 1.010-6.250), university students (OR=2.598, 95%CI: 1.238-5.450) and civil servants (OR=2.733, 95%CI: 1.193-6.263). There were 228 participants with willingness to continue to participate in volunteer blood donation with regular apheresis platelets (96.61%), and 215 participants with a history of participating in volunteer blood donation surveys (91.10%). Internet was the predominant route for acquiring knowledge about apheresis platelet donation (41.10%), and “think that blood donation is a meaningful thing” was the predominant cause of participating in volunteer apheresis platelet donation (33.05%). Conclusions There is a high proportion of regular blood donation among civil servants and university students.

Objective To study the relationship between the level of fluoride in urine and the prevalence of dental caries in children before and after the defluoridation, and to provide a basis for assessment of the effects of defluoridation projects and for control of dental caries.Methods Wamiao Village, in Jiangsu Province, a formerly severe endemic fluorosis area, was selected to carry out the study.All children aged 8-13 years old before the defuoridation were investigated from September to November in 2002, and children 8-10 years old who were born after the defluoridation were investigated from September to November in 2013.Urinary fluoride level of the children and the prevalence of dental caries were determined, and their relationships were analyzed.The urina sanguinis samples of children were collected in the morning, and the urine fluoride level was tested using the fluoride ion selective electrode.Dental caries diagnoses was referenced to Dental Caries.Results Totally children's urine samples were 236 and 68 respectively before and after defluoridation.Urinary fluoride level of the children was significantly decreased from (3.53 ± 1.81)mg/L (before defluoridation) to (1.39 ± 0.66)mg/L (after defluoridation, t =9.506, P ＜ 0.01);the prevalence of dental caries was increased from 52.73％ (29/55, before defluoridation) to 63.24％ (43/68, after defluoridation), however, the difference was not significant (x2 =1.383, P ＞ 0.05).The DMFT increased from 1.18 (before defluoridation) to 1.68 (after defluoridation), and the epidemic levels of dental caries were all at lower levels.The relationship between urine fluoride level and the prevalence of dental caries as well as the DMFT before defluoridation was a U-shape dose-response curve;which was gone after defluoridation.Conclusions The urinary fluoride level is significantly decreased after defluoridation for 10 years, the prevalence of dental caries is increased but not significantly.The results of this study indicate that the measure of fluoridation to prevent dental caries needs to be further validated.

AIM: Tongxinluo had showed effects of inducing proliferation and differentiation on neural stem cells in previous observations on animal models. The experiment would investigate the time-effect and dose-effect relationship of Tongxinluo on neurocyte lineage development from rat embryonic neural stem cells (NSCs). METHODS: Experiments were performed at the Laboratory of Functional Science of Southern Medical University from June to October 2007. ①Pregnant rats of 12-14 days were provided by Experimental Animal Center of Southern Medical University. The experiment was performed in strict compliance with all standards for ethics. Tongxinluo contained ginseng, leech, scorpion, ground beetle, centipede, pellicula cicadae, red peony root, borneol and so on. Tongxinluo was produced by Shijiazhuang Yiling Pharmaceutical Co., Ltd., No. Z19980015. Preparation of serum containing Tongxinluo: 1 g/kg per day Tongxinluo suspension was given by lavage in rats of high-dose Tongxinluo group, and 0.5 g/kg per day in rats of low-dose Tongxinluo group. 7 days later, sera were extracted from all the rats and sterilized with filtrating, followed by separate-loading and cryopreservation at –70 ℃. ②NSCs cultured were separated from 12th-14th day SD rat embryo. The third passage of cells was treated with serum containing Tongxinluo in the high-dose and low-dose Tongxinluo group. Rats were treated with common serum as a control. ③Proportion of various kinds of nerve cells was observed by immunofluorescence staining at days 1, 3 and 7. RESULTS: ①At day 1, there was few cells labeled with 5-bromodeoxyuridine (BrdU) (+) and glial fibrillary acidic protein (GFAP) (+) in high-dose Tongxinluo group, with rest cells all labeled with BrdU (+) and Nestin (+). While in low-dose Tongxinluo group and control group, all cells were BrdU(+)Nestin(+). ②At day 3, the ratios of cells labeled with Nestin (+), ?tubulin (+) or GFAP (+) differentiated with each other (P 0.05). ③The ratios of NSCs labeled with nestin (+) decreased soon after administered Tongxinluo, and increased after that. The ratios of ?tubulin (+) cells increased in the high-dose Tongxinluo group while increased firstly then decreased in the low-dose Tongxinluo group and control group. The ratios of GFAP (+) cells increased in the low-dose Tongxinluo group and control group while increased firstly then decreased in the high-dose Tongxinluo group. The ratios of GalC (+) cells had no remarkable change in the high-dose and low-dose Tongxinluo group, but increased significantly at day 7 in the control group. CONCLUSION: Tongxinluo may induce proliferation and neuronal differentiation of rat embryonic NSCs with time-effect and dose-effect relationship. The more Tongxinluo is administered, the more significant and permanent effects will be achieved.

BACKGROUND:Menstrual blood-derived mesenchymal stem cels are proved to be a new type of adult stem cels. To date, little is reported on the methods of establishing this kind of cel lines systematicaly.
OBJECTIVE:To isolate, culture, identify and finaly establish menstrual blood-derived mesenchymal stem cel lines.
METHODS:Menstrual blood-derived mesenchymal stem cels were isolated from the menstrual blood of healthy female donors and cultured accordingly using density gradient centrifugation method. Cel morphology and proliferation characteristics were observed, and proliferative ability was detected by cel counting kit-8 assay. Cels were colected periodicaly for karyotype analysis. Expressions of cel surface antigens such as CD34, CD38, CD44, CD45, CD73, CD90, CD105 and Oct-4 were analyzed using flow cytometry. Cel pluripotency were identified by inducing adipogenesis and osteogenesis in vitro.
RESULTS AND CONCLUSION:We successfuly established four menstrual blood-derived mesenchymal stem cel lines from the menstrual blood of six healthy volunteers. The cultured cel lines had a typical spindle shape and kept normal 46, XX karyotype. The cels grew rapidly and entered into the logarithmic growth phase at 48 hours after passaging. The results of flow cytometry showed that menstrual blood-derived mesenchymal stem cels were positive for CD44, CD73, CD90, CD105 and Oct-4, while negative for CD38, CD34 and CD45. Menstrual blood-derived mesenchymal stem cels were positive for oil red O staining at 22 days after adipogenic induction and also positive for alizarin red staining at 2 weeks after osteogenetic induction. In conclusion, our study suggests that menstrual blood-derived mesenchymal stem cels with stable karyotype and pluripotency can proliferate quickly, and we can provide a new resource for stem cel therapy and regenerative medicine through establishing the menstrual blood-derived mesenchymal stem cel lines.