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 CNTF gene expression and expressive variety during postnatal development in rat spinal cord. Methods The spatial expression of CNTF mRNA in spinal cord was examined by in situ hybridization with dig\|CNTF cDNA probe.Reverse transcription\|polymerse chain reaction(RT\|PCR),as hemi\|quantitative method was used to investigate the expressive variety of CNTF mRNA levels in spinal cord during postnatal development. Results Hybridized signals of CNTF mRNA was found only in part of glial cells on the peripheral region of the ventral and lateral white funiculi of spinal cord,but could not be detected in the gray matter of spinal cord.The expression of CNTF mRNA in spinal cord appeared with a lower level on the first postnatal day.It increased significantly at postnatal 15th days,the expression of CNTF mRNA reached the peak at 30th days,and it began to decrease on the 60th days. Conclusion This study indicated that CNTF mRNA might be expressed in part of glial cells in the white matter.There was expression in spinal cord on the 1st day after birth and the expression levels of CNTF mRNA possessed the changeable character during spinal cord development.

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

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.

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.

Red blood cell distribution width is an index reflecting the size heterogeneity of circulating red blood cells, which is usually used for diagnosis and treatment of blood system diseases.In recent years, more and more evidences show that the increase of red blood cell distribution width level is closely related to the occurrence, development and prognosis of cardiovascular diseases.We focused on the relationship between red blood cell distribution width and atrial fibrillation (AF) and its possible mechanism, in order to provide some reference value for the clinical treatment of AF.