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.

In order to investigate the characteristics of transdermal delivery of ferulic acid under the treated of microneedle arrays and the influence on permeability of rat skin capillaries, improved Franz-cells were used in the transdermal delivery experiment with the rat skin of abdominal wall and the length of microneedle arrays, different insertion forces, retention time were studied in the influence of characteristics of transdermal delivery of FA. The amount of FA was determined by HPLC system. Intravenous injection Evans blue and FA was added after microneedle arrays treated. Established inflammation model was built by daubing dimethylbenzene. The amount of Evans blue in the rat skin was read at 590 nm wavelength with a Multiskan Go microplate reader. Compared with passive diffusion group the skin pretreated with microneedle arrays had a remarkable enhancement of FA transport (P <0.01). The accumulation of FA increased with the enhancement of insertion force as to as the increase of retention time. Microneedle arrays with different length had a remarkable enhancement of FA transport, but was not related to the increase of the length. The research of FA on the reduce of permeability of rat skin capillaries indicated that the skin pretreated with microneedle arrays could reduce the content of Evans blue in the skins of rat significantly compared with the untreated group. The permeation rate of ferulic acid transdermal delivery had remarkable increase under the treated of microneedle arrays and the length of microneedle arrays ,the retention time so as to the insertion force were important to the transdermal delivery of ferulic acid.
Administration, Cutaneous ; Animals ; Coumaric Acids ; administration & dosage ; pharmacokinetics ; Male ; Needles ; Rats ; Rats, Sprague-Dawley ; Skin Absorption

Objective To study the mechanism of chemotherapy resistance caused by interleukin-6 (IL-6) in ovarian cancer cells and its related signal pathways. Methods Ovarian cancer cell lines A2780(IL-6 receptor positive, while non-IL-6-expressing and cisplatin/paclitaxel-responsive) and SKOV3 cell lines( overexpressing of IL-6 receptor and IL-6 and cisplatin/paclitaxel-resistant) were suitable models for this study. The effect of exogenous (a short period of treatment with recombination IL-6) and endogenous IL-6(by transfecting with plasmid encoding for sense IL-6 ) in A2780 cells or deleting of endogenous IL-6expression in SKOV3 cells (by transfecting with plasmid encoding for antisense IL-6) on the sensitivity to cisplatin and paclitaxel was investigated. Meanwhile, the mechanism of chemotherapy resistance caused by IL-6 in ovarian cancer cells and its related signal pathways were also analyzed. Results We found that both exogenous and endogenous IL-6 induce cisplatin and paclitaxel resistance in non-IL-6-expressing A2780 cells (the resistance multiple to cisplatin and paclitaxel was: exogenous, 6. 25 and 7.31; endogenous, 7. 13 -8. 34 and 7. 61 - 10. 70), while deleting of endogenous IL-6 expression in IL-6-overexpressing SKOV3 cells promotes its sensitivity to anticancer drugs ( the resistance multiple to cisplatin and paclitaxel was 0. 15 and 0. 10, 0. 10 and 0. 08). IL-6 significantly up-regulated the expression levels of mRNA and protein of drug resistance-associated genes, MDR1 and GST-π, and apoptosis-inhibiting genes, bcl-2, bcl-xL and XIAP in a dose-dependent manner in A2780 cells. In accordance with this finding, the mRNA and protein levels of MDR1 and GST-π enhanced in sense IL-6-transfected A2780 cells, and reduced in antisense IL-6-transfected SKOV3 cells compared with the corresponding parental and control vector-transfected cells, which had no difference. It was found that PD98059 [ mitogen-activated protein kinase-extracellular signalregulated kinase (MEK) inhibitor ] and wortmannin [ phosphatidylinositol 3-kinase (PI3K) inhibitor ]significantly antagonized IL-6-induced phosphorylation of extracellular signal-regulated kinase ( ERK ) and protein kinase B (Akt), respectively, and both of them blocked IL-6-induced cisplatin and paclitaxel resistance and the inhibitory effects of PD98059 and wortmannin were dependent on its concentration.Conclusions These data suggest that IL-6-induced chemoresistance may be associated with increase of both drug resistance-associated genes ( MDR1 and GST-π) and apoptosis-inhibiting genes ( bcl-2, bcl-xL and XIAP), and activation of MEK/ERK and PL3K/Akt. Therefore, modulation of IL-6 expression or its related signaling pathway may be a promising strategy of treatment for drug-resistant ovarian cancer.

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 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.

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.