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 explore the effect of radiofrequency catheter ablation( RFCA) on cardiac reverse remode-ling and improvement of life quality in cardiomyopathy patients with paroxysmal atrial fibrillation( PAF) . Methods 95 cardiomyopathy patients with PAF were enrolled in our study and divided into two groups. RFCA group:62 patients received circumferential pulmonary vein isolation, ( left ventricular end-diastolic ) LVEDD ≥55 mm (male), LVEDD ≥50 mm (female); Drug group:33 patients were treated with drug for controlling heart rate ( resting heart rate around 60~80 bpm, heart rate during daily activity <100 bpm) . 72 hours after admission or 6 months after surgery in RFCA group, when the heart rate returned to normal or 6 months after treatment in Drug group, Short-Form36(SF-36) was used to evaluate the quality of living in the patients respectively; transthoracic echocardiography was performed in sinus rhythm;LAD, LVEDD and LVEF of the patients were measured. Results in RFCA group, LAD and LVEDD of 62 patients reduced and LVEF increased in 6 months after surgery statisti-cally significant(P<0. 05). In Drug group, 6 months after treatment, LAD and LVEDD of 33 patients increased ( P<0. 05 ) , without significant change in LVEF. There was no statistical significance in psychological health, physical function and general health perceptions, but there was significant improvement in social function and phys-ical function,affective state, physical role and energy in both RFCA group and drug group (P<0. 05), and it was more obvious in RFCA group(P<0. 05). Conclusion RFCA can reverse cardiac structural remodeling via sinus rhythm maintenance and improve the quality of life in cardiomyopathy patients with PAF.

Female ; Humans ; Infant ; Mitochondrial Proton-Translocating ATPases ; deficiency ; genetics ; Mutation

Objective To evaluate the prognostic value of dynamic delta model of end stage liver disease(MELD)in patients with decompensated liver cirrhosis.Methods Ninty-seven patients with decompensated liver cirrhosis were enrolled in the study and followed for 1 year followed up.Child-Turcotte- Pugh(CTP)score and MELD score were calculated twice for each patient on the first day of admission and one month later.The difference between two MELD scores represented the delta MELD.The predictive value related with delta MELD,MELD and CTP scores was determined by the area under receiver operating characteristic(ROC)curve.Results Ten patients died within 3 months,whose delta MELD(3.23?2.77) were higher than those of survivors(0.15?0.39)(P

Epiglottis ; pathology ; Humans ; Palatine Tonsil ; Tissue Adhesions ; diagnosis ; Tonsillar Neoplasms ; radiotherapy

Objective To observe the expression of Frizzled protein in mouse tooth germ at bell stage and explore the possible function of Wnt/Frizzled signal molecules. Methods Mouse embryos at 17 and 19 days of gestation (E17 and E19) as well as mice postnatal day 2 (PN2) were employed in our study. The frozen sections of the first lower molar were made and indirect immunofluorescence technique was carried out on the sections. The location of Frizzled protein was observed under the fluorescence microscope. Results Frizzled protein was expressed weakly in the inner- and outer- dental epithelium at E17. At E19,the positive staining was found to distribute at the summit of both pre-ameloblasts and pre-odontoblasts. At P2,Frizzled protein was expressed strongly at the summit of ameloblasts and odontoblasts. Conclusion Wnt/Frizzled signal molecules may be involved in the differentiation of ameloblasts and odontoblasts.

Objective To explore the expression of thioredoxin reductase (TrxR) mRNA and protein,as well as the relationship between TrxR and myeloid leukemia.Methods Bone marrow samples from 20 acute myeloid leukemia (AML),20 chronic myeloid leukemia (CML),and 20 healthy persons as normal control group were collected.The human non small cell lung cancer A549 cells were selected as the positive control group.The expression of TrxR mRNA was detected by real-time quantitative polymerase chain reaction.The expression of TrxR protein level was detected by Western blot.Results The relative quantitation expressions of TrxR mRNA were 6.751 (13.459),4.321 (11.389),18.477 (2.089),1.045 (0.467) in AML,CML,positive control and normal control group,were 17.814±3.979 and 4.860±1.550 in the primary diagnosed/relapse group and complete remission (CR) group of AML patients,and were 19.552 (5.758) and 3.459 (2.047) in the primary diagnosed and treatment group of CML patients.The expression levels of TrxR mRNA in AML,CML and positive control group were significantly higher than that in normal control group (H =43.978,P ＜ 0.001).For AML patients,the expression levels of TrxR mRNA in the primary diagnosed/relapse group,CR,positive control group were significantly higher than that in normal control group (F=246.793,P ＜ 0.001),and the difference between the primary diagnosed/relapse group and positive control group was no significant (P ＞ 0.05).The expression of TrxR mRNA were higher in the primary diagnosed/relapse group than that in CR group,and that in CR group than that in normal control group (both P ＜ 0.05).For CML patients,the expression levels of TrxR mRNA in the primary diagnosed,treatment,positive control group were significantly higher than that in normal controls (H =38.222,P ＜ 0.001),the difference between that in the primary diagnosed and that in positive control group was no significant difference (P ＞ 0.05),but the expression of TrxR were significantly higher in the primary diagnosed group than that in treatment group,and that in treatment group than that in normal controls (both P ＜ 0.05).The expression of TrxR mRNA in the primary diagnosed/relapse group of AML and the primary diagnosed group of CML showed no significant difference (P ＞ 0.05).The positive levels of TrxR protein were 0.679,0.606,0.877 and 0.095 in AML,CML,positive control and normal control group.TrxR protein was highly expressed in myeloid leukemia patients.Conclusion The expression levels of TrxR mRNA and protein are increased in myeloid leukemia.The low expression of TrxR in normal hematopoietic stem cells is highly expressed in myeloid leukemia,and is downgraded after treatment,which may be used as one of the parameters to diagnosis,effect and prognosis of myeloid leukemia.

Objective To learn the psychological needs of the hospital staff in terms of their life,medical care and psychological health.Methods 2910 hospital staff were interviewed with questionnaires and the outcomes analyzed with x2 test and descriptive analysis.Results 96.9％of the surveyed found themselves in need of psychological counseling; considerable consulting needs of the staff; most of them turn to friends to complain instead of their leaders.Conclusion The psychological counseling should be enhanced to build effective communication channels and ease stress of the staff.

Objective To compare the efficacy of alfentanil and remifentanil in minimizing the propofol injection pain. Methods A total of 175 adult female patients undergoing gynecological procedures with general anesthesia were randomly divided into four groups.Patients received alfentanil 1mg(2 mL,AL group,n=43),remifentanil 0.01 mg(2 mL,REM1 group,n=43),remifentanil 0.02 mg(2 mL,REM2 group,n=45) or normal saline(2 mL,control group,n=44) 30 seconds prior to propofol administration.Visual analogue scale(VAS) was employed to evaluate the subjective feelings of pain due to propofol injection,and adverse effects were recorded. Results One patient in REM2 group and one patient in control group were excluded due to difficulty in venous catheterization.The injection pain in AL group,REM1 group and REM2 group was significantly less severe than that in control group(P

Background Mesenchymal stem cells (MSCs) have been applied in basic and clinical researches of organ transplantation.Our previous study showed that intravenous injection of MSCs prolonged corneal allograft survival in rat.However,the effect of local administration of MSCs on corneal allograft rejection remains unclear.Objective The aim of this study was to investigate the effect of subconjunctival injection of MSCs on corneal allograft rejection in rat model of keratoplasty.Methods MSCs were isolated and cultured from femur and tibia bone marrow of clean Wistar rats,and then the cells were identified by induced differentiation of osteoblast and adipocyte.The third generation of MSCs was used in subsequent experiment.Allogenic penetrating keratoplasty was performed with the bilateral corneas of 20 Wistar rats as donor grafts and the right eyes of 40 Lewis rats as recipients.PBS 0.1 ml containing 2 × 106 MSCs and 0.1 ml PBS only was subconjunctivally injected immediately and postoperative 3 days respectively in randomized two groups,and another 6 normal Lewis rats served as the normal control group.Corneal rejection response was evaluated under the slit lamp after surgery based corneal opacity,edema and neovascularization,and the grafts were scored according to the criteria of Larkin.The corneal samples were extracted from 12 rats of the PBS control group and the MSCs group separately 10 days after surgery.The relative expressions of Th1 cytokines (interferon-γ [IFN-γ] mRNA and interleukin-2 [IL-2] mRNA) and Th2 cytokines (IL-4 mRNA and IL-10 mRNA) were detected by real-time quantitative PCR.Protein levels of IL-4 and IL-10 proteins in the corneas were assayed by ELISA.All experimental protocols involving rats were approved by the laboratory animal care and use committee of the Tianjin Medical University and treated with the ARVO statement for the use of animals in ophthalmic and vision research.Results The cells grew well with the orange stain for alizarin red in differentiated the osteoblasts and red stain for Oil red O in differentiated adipocytes.The survival time of corneal graft in the MSCs group was (11.8±1.6) days,it was significantly longer than (9.6±1.4) days in the PBS control group (P=0.004).The levels of IL-4 mRNA and IL-10 mRNA in the MSCs group were significantly higher than those in the PBS control group (both at P =0.00);while the levels of IFN-γ mRNA and IL-2 mRNA were not significantly different between the groups (both at P＞0.05).The IL-10 protein contents were (22.74 ±7.06),(68.40±12.83) and (215.41 ±44.66)pg/ml in the normal control group,PBS control group and MSCs group,showing significant difference among the three groups (F =55.06,P =0.00) and a significant increase in the MSCs group compared with the PBS control group and the normal control group (both at P ＜ 0.05).Conclusions Subconjunctival injection of MSCs prolongs the survival time of cornea allograft in penetrating keratoplasty probably by modulating the balance between Th1 and Th2 cytokines,especially by up-regulating Th2 cytokines.