Objective To explore the effects of early growth response gene-1 (Egr-1) on bone marrow mesenchymal stem cells (BMSC) proliferation and osteogenic differentiation,which is aimed at providing new molecular targets for the treatment of osteoporosis.Methods Bone marrow was collected from adult men and the BMSCs were cultured primarily and observed by microscope.Meanwhile,flow cytometry was used for BMSCs phenotypic identification;After transfection of pcDNA3.1/Egr-1 into BM SCs,the level of BMSCs proliferation was determined by MTT respectively on the 2 d,4 d and 6 d;On the 7 d after transfection,the ALP activity assay was used for testing the ALP activity in BMSCs.And then,alizarin red S-calcium kit was used for measuring the calcified knots respectively on the 7 d,14 d and 21 d;On the 21 d after transfection,real-time qPCR and Western blotting were used respectively for measuring the expression of mRNA and protein of Egr-1,Runx2 and NDRG1;Further,BMSCs were transfected with Egr-1 siRNA,and the content of calcium nodules,ALP activity,the expression of Egr-1,Runx2 and NDRG1 were detected as above methods.Results The cells cultured in vitro showed high level of CD90 and CD29 and very low level of CD34 and CD45,which is accorded with the characteristic of BMSCs.The pcDNA3.1/Egr-1 transfection for BMSCs had no effect on cells prolifera tion.However,the calcified knots,ALP activity and the expression of Egr 1,Runx2 and NDRG1 were increased after transfection of pcDNA3.1/Egr-1 for BMSCs.In addition,Egr-1 siRNA showed the opposite effect with pcDNA3.1/Egr-1 transfection for BMSCs.Conclusion Egr-1 induces osteogenic differentiation of BMSCs by promoting NDRG1 but has no effects on proliferation of BMSCs.

Objective To study the effect of varicocele(VC) on the expressions of hypoxia-inducible factor-1?(HIF-1?) and apoptosis-associated gene Bax in germ cells of adolescent rats,and investigate the mechanism of the infertility resulting from varicocele. Methods The varicocele model was established by partial ligation of the left renal vein in adolescent male Wistar rats. A total of 25 rats were randomly divided into 2 groups,namely varicocele group(n=15) and sham operation group (SOG,n=10). All the animals were killed 30 d after operation. The left testis was collected and the expression of HIF-1? and Bax was detected by immunohistochemical method. Results The expression of HIF-1? and Bax in left testis of varicocele group was significantly higher than that of sham operation group(P

Objective To explore the proliferative and apoptotic effect of 3 different siRNAs targeting survivin in human bladder cancer cell line EJ28. Methods Three siRNAs targeting survivin and 1 fluorescence-labeled siRNA as a negative control were chemically synthesized. Transfetion efficiency was observed under fluorescence microscope after transfecting the fluorescence-labeled siRNAs. EJ28 cells were divided into 6 groups: siRNA164 group, siRNA167 group, siRNA389 group, and negative control group, lipofectin group and cell control group. The proliferation of EJ28 cell was detected by MTT assay at 24, 48 and 72 h after the transfection. Apoptotic rate was detected by Annexin V-FITC-FCM assay at 48 h.Results All the sequence-specific siRNAs targeting survivin efficiently suppressed the proliferation of EJ28 cells. siRNA 164 had the most powerful effect because of its highest inhibitory rate of (41.32?2.54)% and the highest apoptotic rate of (13.20?0.25)% at 48 h after transfection. Conclusion RNAi targeting survivin has a potential value in the gene therapy of bladder cancer.

Objective To study the expression of major histocompatibility complex(MHC)on muscle biopsy specimens of idiopathic inflammatory myopathies(IIM),and assess diagnostic value of MHC in IIM. Methods Forty-five patients with IIM(19 polymyositis and 26 dermatomyositis)were selected for this study.Thirty healthy subjects were included as controls.Immunohistochemical staining was applied to identify the expression of HLA-A/B/C and HLA-DR on muscle biopsy-specimens in polymyositis (PM)/deFmatomyosiris (DM) patients and healthy controls.Results HIA-A/B/C antigens were expressed in muscle fibers in 18 out of 19 PM patients(95%),24 out of 26 DM patients(92%) and 3 out 0f 30 healthy controls(10%) respectively.The positive expression rate of HLA-DR in PM,DM patients and healthy controls were 84%, 81%,13%,respectively.The expressions of HLA-A/B/C and HLA-DR were significantly increased in PM and DM patients than those in healthy controls(both P<0.05),but no significant differences were found between PM and DM groups(both P>0.05).No significant correlations were demonstrated between the over-expression of HLA-A/B/C or HLA-DR and the extent of inflammatory infiltrations,muscle damage or clinical features in PM/DM groups (all P values>0.05).Conclusion MHC-Ⅰ and MHC-Ⅱover-expression in muscle fibers are the early events in PM and DM,and may occur in the absence of lymphocyte infihration and muscle damage.Immunostaining for MHC-Ⅰ and MHC-Ⅱcan be used as a routine test in the diagnosis of PM and DM.

Objective To explore the impact of chronic cough on children’s life quality, and to observe their life quality after drugs and psychological intervention. Methods One hundred 9 to 12 years old children with chronic cough were randomly selected. The drugs and psychological intervention were administrated. The children had been follow-up. The children’s quality of life was assessed by“Inventroy of Subjective Life Questionnaire”before and after treatment. Meanwhile 100 healthy children were randomly selected as a control group. Results With the prolonged treatment, the recovery rate and effective rate in children with chronic cough increased. Before the treatment, the scores of family life, peer interaction, self cognitive, experience of depression, experience of anxiety, cognitive and emotional component, and overall satisfaction were significantly lower in children with chronic cough than those in healthy children (P<0.05). After the treatment, the scores of family life, peer interaction, self cognitive, experience of depression, experience of anxiety, physical experience, cognitive and emotional component, and overall satisfaction were significantly improved in children with chronic cough (P<0.05), even the scores of physical experience and emotional component were significantly higher in children with chronic cough than those in healthy children (P<0.05). Conclusions The quality of life in children with chronic cough decline, however, drug and psychological intervention can improve their quality of life.

Objective:To establish the quality control of Zhiqikang capsules. Methods:TLC was used to identify Gastrodia tuder halimasch, rhubarb and Astragalus mongholicus in the preparations. A spectrophotometry method with 3, 5-dinitrosalicylic acid (DNS) was used to measure the polysaccharide content in Zhiqikang capsules. A spectrophotometry method with Forint phenol method ( Low-ry) was used to measure the peptide content in the capsules. Results:The linear range of polysaccharide was obtained between 6. 412 and 32. 060μg·ml-1(r=0. 999 5), the average recovery was 95. 86% and RSD was 0. 86%. The linear range of peptide was ob-tained between 0.059 7 and 0.298 4 mg·ml-1(r=0.999 0), the average recovery was 100.3% and RSD was 1.88%(n=6). Conclusion:The assay method is simple and accurate in the quality control of the preparations.

Objective To summarize the clinical characteristics of primary pigmented nodular adrenocortical disease (PPNAD).Methods Eight patients with PPNAD from 2001 to 2009 in PUMC Hospital were reviewed,their clinical data were collected.Results PPNAD often occurred in adolescents.62.5% of patients with PPNAD were complicated with Carney complex(CNC).In addition to general features of Cushing's syndrome,amenorrhea and growth retardation in stature were very frequent in clinical manifestations of PPNAD.Plasma ACTH was undetectable,circadian rhythm of plasma cortisol was disappeared,glucocorticoid excretion was increased paradoxically during the dexamethasone suppression test in 50% patients with PPNAD.Adrenal imaging from 75% patients revealed normal-sized adrenal glands or suspectable micronodular changes.Adrenal pathologic analysis revealed numerous brown cortical nodules containing lipofuscin pigmentation.Unilateral adrenalectomy may relieve symptoms of Cushing's syndrome,but plasma ACTH and circadian rhythm of plasma cortisol were difficult to recovere.Hypercorticoidism might re-occure after unilateral adrenalectomy.Conclusion PPNAD should be bewared in ACTH independent Cushing's syndrome patients without apparent adrenal mass,and CNC should be screened and followed up.

AIM:To establish the GC fingerprint of Huoxiang Zhengqi Solution.METHODS:The volatile constituents of Huoxiang Zhengqi Solution were analyzed by capillary GC with FID detector using hydrodistillation and hexane extraction under n-hepladecane used as the reference substance.RESULTS:GC fingerprint of Huoxiang Zhengqi Solution,16 common peaks were established on the basis of systematic methodology after 10 batches of samples were tested.Variation in the relative retention time of 16 identified common peaks were within 0.5% range.CONCLUSION:The analytical method for Huoxiang Zhengqi Solution is precise and reliable.The research would be helpful to offer an effective pattern for quality control of Huoxiang Zhengqi Solution.

Objective·To study the change of circulating endothelial progenitor cells (cEPCs) in acute ischemic stroke (AIS) patients within one week after attack,and the correlation of cEPCs with the prognosis.Methods·Ninty-two patients with AIS (AIS group) and 20 patients with risk factors (Risk group) were recruited.The proportion of cEPCs (CD34TKDR+ cells) in peripheral blood mononuclear cells of AIS patients was measured by flow cytometry (FCM) on the first day of admission and the seventh day after attack.Functional recovery was assessed by modified Rankin Scale (mRS) on the 90th day after onset.The cEPCs percentages of AIS patients with different mRS were compared to analyze their correlation.Results·Compared with Risk group,cEPCs percentage of AIS group on the 1st day of admission was lower (P=0.016).In AIS group,compared with poor prognosis group (mRS＞2),eEPCs percentage of good prognosis group on the 7th day after onset (mRS ≤ 2) elevates (P=0.002).The result of multiple linear regression showed that cEPCs percentage on the 7th day after onset was positively correlated with mRS on the 90th day (t=4.608,P=0.011).Conclusion·The percentage of cEPCs in peripheral blood of AIS patients decreases significantly during the acute phase.The percentage on the 7th day after onset is correlated with prognosis of AIS patients.

AIM:To establish the GC fingerprint of Huoxiang Zhengqi Solution.METHODS:The volatile constituents of Huoxiang Zhengqi Solution were analyzed by capillary GC with FID detector using hydrodistillation and hexane extraction under n-heptadecane used as the reference substance.RESULTS:GC fingerprint of Huoxiang Zhengqi Solution,16 common peaks were established on the basis of systematic methodology after 10 batches of samples were tested.Variation in the relative retention time of 16 identified common peaks were within 0.5% range.CONCLUSION:The analytical method for Huoxiang Zhengqi Solution is precise and reliable.The research would be helpful to offer an effective pattern for quality control of Huoxiang Zhengqi Solution.