OBJECTIVETo explore the role of the basic fibroblast growth factor (bFGF) in the directional differentiation of bone marrow mesenchymal stem cells (BMSCs) into Leydig cells.

METHODSAfter purification and identification, we inoculated the third-generation BMSCs of SD rats onto a six-orifice board and then randomly divided them into groups A (normal saline control), B (human chorionic gonadotropin [hCG] + platelet-derived growth factor [PDGF] induction), C (hCG + PDGF + 5.0 ng/ml bFGF induction), D (hCG + PDGF + 10.0 ng/ml bFGF induction), and E (hCG + PDGF + 20.0 ng/ml bFGF induction). On the 7th, 14th and 21st day of induction, we observed the morphological changes of the cells and measured the level of testosterone (T) and expression of 3 beta hydroxy steroid dehydrogenase (3β-HSD) in the supernatant by immunofluorescence staining.

RESULTSAfter induction, the BMSCs of groups B, C, D, and E exhibited microscopic features of enlarged size, inter-connection, long-shuttle or irregular shape, adherent growth, and large round nuclei, all characteristic of Leydig cells. With the prolonging of time and enhanced concentration of bFGF, gradual increases were observed in the T level and the count of 3β-HSD-positive BMSCs in the four induction groups, with statistically significant differences between group B and groups C, D, and E (P < 0.05), as well as between group C and groups D and E (P < 0.05), but not between D and E (P > 0.05).

CONCLUSIONThe bFGF has an obvious promoting effect in the in vitro induced differentiation of rat BMSCs into Leydig cells.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Differentiation ; Cells, Cultured ; Chorionic Gonadotropin ; metabolism ; Fibroblast Growth Factor 2 ; pharmacology ; Humans ; Leydig Cells ; cytology ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Testosterone ; metabolism

Objective To study the therapeutic efficacy of Rex shunt in treating pediatric patients with prehepatic portal hypertension (PHPH).Methods Five children with PHPH who were admitted from October 2014 to May 2015 were reviewed.There were three boys and two girls,with age ranging from 50 to 95.5 months [(75.8 ± 1.9) months].They all suffered from recurrent upper gastrointestinal (GI) bleeding.Their red blood cell (RBC),white blood cell (WBC) and platelet (PLT) counts were decreased,but laboratory findings revealed no liver dysfunction.Ultrasound and CT scan diagnosed cavernous transformation of portal vein (CTPV) and splenomegaly.The mean splenic length was (42.8 ± 8.2) cm.Indirect portal venography revealed patent left portal vein.All patients underwent Rex shunt and were followed up for 3 ～ 7 months.Results The mean duration of operation was (566.0 ± 39.7) min.Intraoperative bleeding varied from 10 to 50 ml.The portal pressure significantly decreased after surgery from [(25.6 ± 1.5) mmHg,1 mmHg =0.133 kPa] preoperatively to (19.2 ± 3.3) mmHg postoperatively (P ＜ 0.05).Portal venography indicated patent left portal vein after the Rex shunt.The postoperative course was uneventful in the five patients with a mean hospital stay of (26 ± 9.3) days.There was no further GI bleeding.The RBC,WBC and PLT counts increased.Ultrasound indicated patent anastomotic stomas and decreased splenic size.Conclusion A Rex shunt in treating patients with PHPH is safe,feasible and efficacious.

Objective To study the influence of the preparation and quality control of Xiaochuan cataplasm on its drug action. Methods Xiaochuan cataplasm was prepared with hydrosoluble polymers matrix. A sequence of tests such as adhesive force test, recontour test, paste-content test, cold-resistance test, heat-resistance test, toxicity and pharmacodynamic action tests were performed on this drug. Results Xiaochuan cataplasm contains a sound volume of medicine, and shows good adhesive property, no sensibilization and stimulation, and notable drug action. Conclusion The results demonstrate that the preparation craft of Xiao-chuan cataplasm is reasonable and practicable., the quality is controllable and the method is a safe and effective transdermal drug delivery system.

Human cytomegalovirus (HCMV) late mRNA expression in megakaryoblast and in turn the pathogenesis of idiopathic thrombocytopenic purpura (ITP) patients with HCMV infection, and effectiveness of ganciclovir were investigated. Colony forming unit-megakaryocytes (CFU-MK) of 46 ITP patients with HCMV infection were incubated from patients' bone marrow mononuclear cells (MNC). Reverse transcriptase-polymerase chain reaction (RT-PCR) was subsequently used for CFU-MK for HCMV-late mRNA detection. Ganciclovir therapy was given to both HCMV-late mRNA positive and negative groups for comparison of therapeutic effectiveness. The results in 19 of 46 CFU-MK culture cells specimens with positive HCMV-DNA by PCR or positive CMV-IgM by enzyme linked immunosorbent assay (ELISA) in the correspondent serum of peripheral blood were positive for HCMV-late mRNA. Sixteen out of 19, patients with positive HCMV-late mRNA CFU-MK had a positive response to ganciclovir. Amongst 27 patients with negative HCMV-late mRNA CFU-MK, only 4 positive responders to ganciclovir therapy were observed. Curative effectiveness of ganciclovir in HCMV-late mRNA positive group was significantly higher than that in HCMV-late mRNA negative group (P<0.01). It was suggested that HCMV could directly infect CFU-MK, which might be one of the mechanisms responsible for HCMV related ITP. The ganciclovir is an effective therapy in resulting in the increases in thrombocyte in the ITP patients whose HCMV- late mRNA was positive in their CFU-MK.

Objective To explore the effectiveness of sandplay therapy combined with family counseling in children with school phobia and its influence of child′ self-consciousness. Methods Integrative sandplay therary with family consulting were used to treat 28 patients with school phobia regularly for 2 months. Sandplay and family consulting therapy were given once a week for 45 minutes . Clinical outcomes were assessed using CGI-GI and Piers-Harris children′s self-consciousness scale before and after treatment as well as 3 months posttreatment. Results Overall response rate was 85%. In addition, the physical appearance and characteristic factor before and after treatment were no significant difference (P>0.05). The rest of the various factors and total score compared with pre-treatment significantly improved (P<0.05). After treatment for 3 months, every factor in self-consciousness of children and total score were no significant difference (P>0.05). Conclusion Integrative sandplay therapy with family counseling has better and long-lasting treatment effect to self-consciousness of children with school refusal.

Objective To generate an autoimmune cholangitis animal model and investigate whether CXCR3 and its ligand were involved in the pathogenesis of primary biliary cirrhosis (PBC). Methods Female C57BL/6 wild-type (WT) mice and CXCR3 knockout (CXCR3-/-) mice were injected with 5 mg/kg of poly I:C intra-peritoneally twice a week for 24 consecutive weeks. Liver specimens were collected to evaluate the degree of cell infiltration. AMA was assayed by ELISA. Differences in pathology were compared between CXCR3-/- mice and wild-type. Student's test was used to assess the differences. Results Anti-mitochon-drial antibody (AMA) and alkaline phosphatase (ALP) level were elevated significantly in the sera of all poly Ⅰ:C injected mice compared with control mice. AMA titers in serum were increased in the poly I:C injected WT mice compared with that of the control mice at week 8, 16, and 24 respectively (0.70±0.41 vs 0.17±0.04,0.48±0.35 vs 0.19±0.07, 0.69±0.44 w 0.20±0.06, P＜0.01). There was no significant difference between AMA titers in the serum of WT PBC mice and CXCR3-/- PBC mice (0.70±0.41 vs 0.56±0.25, 0.48±0.35 vs 0.46±0.35, 0.69±0.44 vs 0.85±0.34). Serum alkaline phosphatase (ALP) levels were raised among the poly I:Cinjected WT mice compared with WT controls (115±33) vs (65±7) U/ml; (119±32) vs (70±9) U/ml; (133±52) vs (77±10) U/ml. The serum ALP levels in CXCR3-/- PBC mice were (106±29), (112±29)and (122±60) U/ml at week 8, 16 and 24 respectively. There was no significant difference in ALP level between WT and CXCR3-/- mice PBC model. Considerable numbers of infiltrating cells were detected at the portal areas 8weeks after poly I:C injection, which progressed up to 24 weeks. At week 24, the interlobular bile ducts were lost and bile-plug were evident. Compared to WT mice model, this results revealed that CXCR3-/- mice, had fewer foci of inflammation and significant reduction in total inflammatory cells in their livers than those of the WT mice at 8, 16 and 24 weeks after injection of poly I:C. At week 24, there were no cholestasis orgranulomas in CXCR3-/- mice of PBC models. Compared to the control model, CXCL10 serum level was increased in PBC model. With the disease progression, CXCL10 serum level was elevated gradually. In comparison with wild mice, CXCR3-/- mice model had a higher serum level of CXCL10. At week 24, there was significant difference of CXCL10 serum level between wild-type mice and CXCR3-/- mice model. Conclusion In conclusion, these findings indicate that, CXCR3-/- mice might have a delayed onset and ultimately could not recruit sufficient effector cells to the liver for inflammation development.

Objective To generate an primary biliary cirrhosis animal model and investigate whether CXCR3 and its ligands were involved in the pathogenesis of primary biliary cirrhosis (PBC).Methods Female C57BL/6 wild-type(WT)mice were injected with 5 mg/kg of poly I:C intraperitoneally twice a week for 24 consecutive weeks.Establishment of PBC was confirmed by liver function test,serom autoantibodies and liver biopsy.Expression of CXCR3 on lymphocytes of liver/spleen and level of CXCL10 in peripheral blood were tested by flow cytometry assay and ELISA.The t-test was used for two group data comparison.Results Anti-mitochondrial antibody was detected in the sera of all poly I:C injected wild type C57BL/6mice.Considerable numbers of inflammatory cells were detected at the portal areas 8 weeks after the initiation of poly I:C injection,which progressed up to 24 weeks.Compared the to control mice,serum level of CXCL10was increased in PBC mice.With the disease progression,CXCL10 serum level was elevated.The level of CXCL10 at 8,16 and 24 weeks in PBC model was 0.28±0.10,0.33±0.19 and 0.27±0.11,which were much higher than those of the control mice.CXCL10 serum level of control mice was 0.07±0.03.0.08±0.05,0.10±0.04 respectively.Compared to control model,the proportion of CXCR3+ positive cells was inereased in the intrahepatic infiltrates of PBC model,mostly on CD8+ T cells.Moreover,the expression of CXCR3 was decreased in CD3+and CD8+splenocytes from PBC model compared with control model.Conclusion CXCR3and CXCL1O may attract T cells to the liver of PBC mice mode in the process of PBC development

Objective To assess the mid-term result of laparoscopy in resection of choledoehal cyst,hepaticojejunostomy and discuss the technical improvement.Methods Retrospectively analyzed the data of 31 patients who had undergone laparoscopic choledochal cyst excision from November 2010 to November 2012.Under laparoscopic guidance,intraoperative cholangiogram was performed,Rorx-Y jejunojejunostomy was performed extracorporeally through umbilical incision,then the dilated bile duct were completely excised and a hepaticojejunostomy was carried out intracorporeally.Results Twenty-nine patients successfully underwent laparoscopic choledochal cyst resection,while 2 patients were converted to open surgery because of severe adhesion.The mean operation time was 260 minutes (range from 200 to 460 minutes).A follow-up of 3 to 27 months were achieved in all patients.Three patients developed early complications:one bile leakage,one incision port hemorrhage and one intestinal torsion.The laboratory tests were normal and no dilation of the bile duct was detected in all patients.Conclusions Laparoscopic total cyst excision with Roux-Y hepaticojejunostomy was effective and safe procedure with satisfied results.

Objective To explore the significance, indicatinos and complications of emergency tracheotomy for patients with severe craniocerbral trauma. Methods The clinical data of 42 patients with severe craniocerbral trauma were retrospectively analyzed. Results Of them, 20 were basically cured (48%), 12 had improvement (29%) ,3 were released from the hospital(7 % ), 1 survived(2 % ) in a vegetative state,and 6 died(14%). Conclu- sion Emergent tracheotomy should be performed on patients with severe craniocerbral trauma to resolve respiratory tract obstruction, improve air current, prevent cerebral anoxia, prevent and treat pulmonary infection, increase the therapeutic efficacy.

Objective To investigate the function of acid pocket in reflux esophagitis. Methods The 15 healthy controls and 24 reflux esophagitis patients were identified by reflux disease questionnaire (RDQ) and gastric endoscopy. The location of subjects' lower esophageal sphincter (LES) was determined by 4 channel esophageal manometry system. Then a single-channel pH electrode was positioned 1 cm below the distal border of the LES to monitor fasting pH for half an hour. After a standard meal, the pH was continuously measured for two hours. Then the electrode was moved to 5 cm above the proximal border of the LES to monitor the dynamic pH for 24 h.Results Acid pocket was found in 16 cases of reflux esophagitis patients(66.67%) and 10 cases of healthy individuals (10/15). Acid pocket occurred earlier in reflux esophagitis group than healthy controls [11.00(4.25-17.00) min vs 30.00(15.50-54.25) min, P＜0.05], and the average pH value was lower [1.84(1.59-2. 19) vs 2.32 (1.96-2.71), P＜0.05]. There was no statistic difference in mean pH value of gastroesophageal junction and the duration of acid pocket before the meal.Conclusion There is abnormal acid reflux in reflux esophagitis patients, and acid pocket with earlier occurrence and lower pH value may relevant to esophageal mucosal impairment.