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 investigate the concentration of 17β-estradiol in bone marrow stem cells (BMSCs) and different tissues of rats, and study the function of estradiol produced in extragonadal sites prelimi-narily. Methods (1) The concentration of 17β-estradiol in lysate of BMSCs and in supernatant cell culture media were detected by Elisa method after rats BMSCs were serum-free cultured in 0 h , 24 h and 48 h respective ly . (2) The tissues of organs were grinded and broken under ultrasonic wave , then washed and weighed. The con-centration of 17β-estradiol in different tissues was detected by Elisa method. Results Compared with the concen-tration of 17β-estradiol at 24 h, the concentration of 17β-estradiol at 48h significantly increased (P < 0.05). The concentration of 17β-estradiol among different tissues were higher, but the difference is not significant (P > 0.05). Conclusions (1) 17β-estradiol can be secreted by BMSCs , and the concentration is proportional to the time to some degree. But whether the extragonadal estrogen can function locally is still unclear. (2) The concentration of 17β-estradiol is non- gonad dependent but whether it is secreted locally remains to be elucidated. The non- gonad estrogen could be the estrogen source of bone metabolism in order to sustain bone health after menopause.

Objective To investigate the expressions and clinical significance of Ki-67,p53,and survivin in esophageal cancer and precancerosis.Methods The expressions of Ki-67,p53,and survivin proteins were detected by immunohistochemical staining in 40 normal esophageal mucosa,136 precancerosis (42 mild atypical hyperplasia,43 moderate atypical hyperplasia,and 51 severe atypical hyperplasia),and 68 esophageal cancer tissues.The correlation of three proteins expressed in esophageal carcinoma tissues was analyzed.Results The positive expression rate of Ki-67 was 0 (0/40)for normal epithelium,35.7％ (15/42) for mild dysplasia,51.2％ (22/43) for moderate dysplasia,74.5％ (38/51) for severe dysplasia,92.6％ (63/68) for squamous carcinoma,respectively.The positive expression rate of p53 protein was 0 (0/40) for normal epithelium,28.6％ (12/42) for mild dysplasia,46.5％ (20/43) for moderate dysplasia,52.9％ (27/51) for severe dysplasia,67.6％ (46/68) for squamous carcinoma,respectively.The positive expression rate of survivin protein was 0 (0/40) for normal epithelium,38.1％ (16/42) for mild dysplasia,55.8％ (24/43) for moderate dysplasia,64.7％ (33/51) for severe dysplasia,89.7％ (61/68) for squamous carcinoma,respectively.Rank correlation analysis showed that abnormal expressions of Ki-67,p53,and survivin were correlated significantly with the pathological grading of the lesions (r =0.637,0.454,0.590,P ＜0.01).The expressions of Ki-67,p53,and survivin were positively correlated in esophageal carcinoma (r =0.407,0.646,P ＜ 0.01).Conclusions The abnormal expressions of Ki67,p53,and survivin were associated with the processes of the esophageal canceration,and the joint detection with three parameters has important clinical value.

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.