OBJECTIVETo study the autophagy activity between rat bone marrow stem cells (BMSCs) neural differentiation in order to explore the mechanism involve in this process.

METHODSBMSCs were passed by 3 generation, then was induced with the revulsant 2% (DMSO) + 200 µmol/L (BHA), NSE expression was detected by immunocytochemical stain, the mRNA expression of autophagy associated genes L3B, Beclinl, Atg5, Atg7, Atg10 were detected by RT-PCR, the autophagy protein LC3B was examined by Western blot and flow cytometry analysis.

RESULTSBMSCs were passed by 3 generation, the purity of BMSCs could reach more than 90%, the morphology of cells were like fibroblasts, after the revulsant 2% DMSO + 200 µmol/L BRA induced, cells were extended long neurites, like nerve cells, positive rate of NSE staining was (83±5) %, RT-PCR results showed that the expression of autophagy associated genes LC3B, Beclinl, Atg5, Atg7 Atg0 were rised after BMSCs neural differentiation, Western blot analysis showed that the LC3B-II protein expression was increased after neural differentiation and the MFI of L3B was highten by flow cytometry.

CONCLUSIONAutophagy is increased after rat BMSC neural differentiation.

Animals ; Autophagy ; Cell Differentiation ; Cells, Cultured ; Flow Cytometry ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Rats

Objective To study the effects of nitric oxide on Ca2+-activated K+ (KCa) channels in the hypothalamus neurons of newborn SD rats. Methods The data were recorded using inside-out or cell-attached configuration of patch-clamp technique and the kinetic changes of KCa channels are compared before and after 100 μmol/L sodium nitroprusside (SNP) was added into the bathing solution. Results When 100 μmol/L SNP was added to the bathing solution in cell-attached configuration, the open probability of the KCa channels increased from (7.3±1.5)% to (40.2±6.5)%, open time from (7.12±1.41) ms to (15.34±3.45) ms, frequency from (11.3±3.5) Hz to (26.6±4.2) Hz. Conclusion NO-cGMP signal pathway could greatly increase the open probability of the channels as a result of both the prolongation of open period and increase of open frequency, but the pathological or physiological roles it may play require further study.

Objective To study the effects of nitric oxide on Ca2+-activated K+ (KCa) channels in the hypothalamus neurons of newborn SD rats. Methods The data were recorded using inside-out or cell-attached configuration of patch-clamp technique and the kinetic changes of KCa channels are compared before and after 100 μmol/L sodium nitroprusside (SNP) was added into the bathing solution. Results When 100 μmol/L SNP was added to the bathing solution in cell-attached configuration, the open probability of the KCa channels increased from (7.3±1.5)% to (40.2±6.5)%, open time from (7.12±1.41) ms to (15.34±3.45) ms, frequency from (11.3±3.5) Hz to (26.6±4.2) Hz. Conclusion NO-cGMP signal pathway could greatly increase the open probability of the channels as a result of both the prolongation of open period and increase of open frequency, but the pathological or physiological roles it may play require further study.

Objective To explore the perinatal outcomes of women with pulmonary hypertension complicating congenital heart disease(CHD).Methods Clinical data of 45 cases of pregnant women with pulmonary hypertension complicating CHD from Apr 1995 to May 2007 were analyzed and they were divided into three groups:29 cases of slight group[pulmonary hypertension of 30 mm Hg(1 mm Hg=0.133 kPa) to 49 mm Hg],8 cases of moderate group(pulmonary hypertension of 50 mm Hg to 79 mm Hg)and 8 cases of severe group(pulmonary hypertension equal to or higher than 80 mm Hg).The types of CHD,cardiac functional status(New York heart association,NYHA),gestational weeks of pregnancy termination,mode of delivery,pregnancy after CHD operation and outcomes of infants were compared between the groups. Results(1)The highest incidence of CHD were atrial septal defect and ventricular septal defect(58%, 26/45).The rate of pregnant women after CHD operation was 29%(13/45),they were mainly in slight group and their NYHA class were in Ⅰ-Ⅱ.(2)The occurrence rate of NYHA class Ⅲ-Ⅳ was 7/8 in severe group.The rate of NYHA class Ⅰ-Ⅱ as 6/8 in moderate group.The rate of NYHA class Ⅰ- was 97%(28 /29)in slight group.(3)The rate of term delivery was 93%(27/29),preterm labor 3% (1/29),abortion 3%(1/29),and the birth weight was(3153?399)g on average in slight group.The rate of term delivery was 5/8,preterm labor occurred in 3 cases in moderate group.The rate of term delivery was 5/8,preterm labor occurred in 2 cases,and iatrogenic abortion in 1 case in severe group.The average birth weight between slight group and moderate or severe group had a significant difference.(4)Caesarean section rate was 78 %(35/45)among all patients.The rate of cesarean section delivery was 76%(22/29)in slight group,6/8 in moderate group,and 7/8 in severe group.(5)The rate of pregnant women who had portent heart failure or heart failure was 24%(11/45),overall maternal mortality was 4%(2/45).Conclusions The higher the pulmonary hypertension,the worse the outcome of the mother and fetus;The pregnant women with good heart function after cardiac operation would have a good perinatal outcome.Cesarean section is more suitable for those women.

Adolescent ; Arrhythmias, Cardiac ; etiology ; Cardiac Catheterization ; Child ; Child, Preschool ; Electrocardiography ; Female ; Heart Septal Defects, Atrial ; therapy ; Humans ; Infant ; Male

Background There have been an abundance of literature on the analysis of the mechanical characteristic of the sclera at the entire seleral level in high myopia.However,some recent studies on high myopia are focused on the mechanical changes of the sclera on a cellular level. Objective This experiment was purposed to study how transforming growth factor-β2(TGF-β2)affected sclerotic desmocytes and the mechanical behaviors of scleral fibroblasts in the posterior part of the eyes in guinea pigs with experimental myopia. Methods Induced myopic animal models were established by wearing-10.00 D concave lens for 30 days in lateral eyes of 2-week-old guinea pigs.The fellow eyes were used as control group.Another 5 matched animals served as normal controls.The scleral fibroblasts of each group were purified with the tissue explant method and passaged for 2 generations in vitro.Cultured cells were identified by immunochemistry with vimentin,desmin,keratin and S-100 antibodies.Different concentrations of TGF-β2(0,1,10,100mg/L)were added into serum-free DMEM for 24 hours,and the viscoelastic properties of scleral fibroblasts were measured by micropipette aspiration technique. Results Compared with the fellow,eyes and normal control eyes,the equilibrium modulus and apparent viscosity in model eyes were significantly higher(P＜0.05).After treatment of TGF-β2,the equilibrium modulus and apparent viscosity in the model group and fellow eyes were positively correlated with the concentrations of TGF-β2(r=0.743,r=0.533,r=0.654,r=0.576,P＜0.05).Following the addition of 1 mg/L TGF-β2 and 10 mg/L TGF-β2,the equilibrium modulus and apparent viscosity of scleral fibroblasts were significantly reduced in model eyes compared with fellow eyes(P＜0.05).No significant difference was found in the equilibrium modulus and apparent viscosity of scleral fibroblasts between model eyes and fellow eyes after treatment with 100 mg/L TGF-β2(P＞0.05). Conclusion TGF-β2 car increase the mechanic indexes in a concentration.dependent manner.1 mg/L,10 mg/L TGF-β2 can lower the equilibrium modulus and apparent viscosity of scleral fibroblasts of normal eye and thus cause more changes in the mechanical behavior of scleral fibroblasts.

OBJECTIVETo analyze the indication of reoperation of thyroid cancer and to explore the timing, surgical pattern of reoperation.

METHODSProtocols of 72 patients underwent reoperation of thyroid cancer from June 2003 to August 2006 were reviewed retrospectively. Causes for reoperation were as follows: residue of the tumor locally as the inappropriate initial operation; local recurrence and cervical lymph node metastasis; before (131)I ablation which differentiated thyroid cancer with distant place metastasis. The reoperation style included residual lobectomy plus isthmus with single tumor below 2 cm, total thyroidectomy in most the other conditions and selective lymph node dissection in finding or suspected cervical lymph node metastasis.

RESULTSThe rate of residual in thyroid confirmed by postoperative pathology was 47.1% (32/68). The rate of residual in cervical lymph node was 81.4% (35/43). The rates of temporary and permanent laryngeal recurrent nerve injury were 5.6% (4/72) and 1.4% (1/72) respectively. The rates of temporary and permanent hypocalcemia were 26.4% (19/72) and 1.4% (1/72) respectively.

CONCLUSIONSInadequate operation, local recurrence and cervical lymph node metastasis of thyroid cancer need revision surgery undoubtedly. The optimal treatment was total thyroidectomy and level VI, VII central compartment lymph node dissection plus intraoperative frozen-section evaluation.

Adolescent ; Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neck Dissection ; Reoperation ; methods ; Retrospective Studies ; Thyroid Gland ; pathology ; surgery ; Thyroid Neoplasms ; pathology ; surgery ; Thyroidectomy ; methods

This study is to investigate the effects of puerarin on the proliferation and differentiation of umbilical cord mesenchymal stem cells (MSCs) into osteoblasts. Umbilical cord MSCs were cultured by tissue adherence and the third passage of cells was used in the experiment. The effect of puerarin on proliferation of umbilical cord MSCs was measured with MTT. The effects of puerarin on umbilical cord MSCs were evaluated by ALP immunohistochemisty and von kossa staining. The OD value decreased with the increase of puerarin concentration. On 7th day, ALP expression of puerarin group was higher than that of control group. On 14th day, ALP staining showed that the positive rate of puerarin group was higher than that of control group. Von kossa staining showed the quantity of calcium nodules was higher in puerarin group than that of control group. Puerarin can promote the umbilical cord MSCs to differentiate into osteoblasts and has an effect on the proliferation of umbilical cord MSCs.
Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Humans ; Isoflavones ; isolation & purification ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; cytology ; Osteogenesis ; drug effects ; Plants, Medicinal ; chemistry ; Pueraria ; chemistry ; Umbilical Cord ; cytology

BACKGROUNDHelical tomotherapy (HT) is a new image-guided intensity-modulated radiation therapy (IMRT). The aim of this study was to evaluate the changes in the target volume and organs at risk (OARs) of patients with nasopharyngeal carcinoma (NPC) during helical tomotherapy.

METHODSForty-three patients with NPC and treated via HT from March 2008 to January 2010 were reviewed retrospectively. Repeated CT scanning and plan adaptation were conducted at the 20th fraction during radiotherapy. The volumetric differences between the two scans were evaluated for nasopharyngeal tumor and retro-pharyngeal lymph nodes (GTVnx), neck lymph nodes (GTVnd), and parotid glands, as well as the axial diameter of the head.

RESULTSThe median interval between the two scans was 25 days (23 - 28 days). The volumetric decrease in GTVnx was 30.1% (median, 29.8%) and in GTVnd 41.6% (median, 45.9%). The variation in the GTVnd volume was correlated with the weight loss of the patient. The volume of the left parotid gland decreased by 35.5% (median, 33.4%) and of the right parotid glands decreased by 36.8% (median, 33.5%). The axial diameter of the head decreased by 9.39% (median, 9.1%).

CONCLUSIONSThe target volume and OARs of patients with NPC varied considerably during HT. These changes may have potential dosimetric effects on the target volume and/or OARs and influence the clinical outcome. Repeated CT scanning and replanning during the HT for NPC patients with a large target volume or an obvious weight loss are recommended.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carcinoma ; Child ; Female ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; radiotherapy ; Organs at Risk ; Radiotherapy, Intensity-Modulated ; Retrospective Studies ; Young Adult

OBJECTIVETo evaluate the feasibility of angiography combined with transthoracic echocardiography (TEE) as a modified management of the transcatheter occlusion of patent ductus arteriosus (PDA).

METHODSForty children with PDA were randomly divided into two groups (n=20 each): observed and control. The control group accepted traditional transcatheter occlusion, and the observed group received a modified management (angiography combined with TEE). The children in the observed group were monitored by realtime TTE.

RESULTSA complete occlusion was acquired by one occlusion operation in each child in the observed group. The TTE demonstrated that the occlusion device was in place, and that the blood flow velocities in the left and right pulmonary artery and the descending aorta were in normal ranges. There were shorter X-ray exposure time, shorter recovering time and less ICU stay time in the observed group than in the control group. The complications associated with blood vessel puncturation occurred in four children from the control group, but none of the observed group had the complications. The total hospitalization cost in the observed group was less than in the control group.

CONCLUSIONSAngiography combined with TEE as a modified management of the transcatheter occlusion of PDA is recommended.

Adolescent ; Cardiac Catheterization ; Cardiac Surgical Procedures ; methods ; Child ; Child, Preschool ; Ductus Arteriosus ; diagnostic imaging ; Ductus Arteriosus, Patent ; diagnostic imaging ; surgery ; Echocardiography ; Humans ; Infant ; Radiography