Objective To study of the effect of traditional Chinese medicine(TCM): Herba radix polygoni multiflori preparata (RPMP) on the cultured melanocytes(MC). Methods Extracts of TMC RPMP treated the cultured melanocytes, and tyrosinase activity, melanin level and proliferation of the cells were determined. Meanwhile, method of micropore filter was used to measure the ability of cell migration. Results TCM RPMP markedly promoted the proliferation and migration of MC in a dose-dependent manner at a range of 0～150?g/ml. Conclusions TCM RPMP can promote the proliferation and migration of MC in vitro. And it suggests that this drug could be used to treat vitiligo.

Objective To explore the impact of high‐load atorvastatin on T cell subsets in patients with unstable angina (UA) after percutaneous coronary intervention (PCI) .Methods One hundred and eighty patients with UA were randomly divided into high‐load atorvastatin group ,ordinary‐load atorvastatin group and routine group ,60 cases in each group .The ratios of CD4+ T cell , CD8+ T cell and the frequencies of CD4+ CD25+ Treg were detected in 3 groups 1 day before PCI and 1 week ,1 month and 6 months after PCI by flow cytometry analysis .Results Different doses of atorvastatin reduced the ratio of CD4+ T cells and in‐creased the ratio of CD8+ T cells and also the frequencies of CD4+ CD25+ Treg after PCI for 1 week ,1 month and 6 months .The longer the time to take atorvastatin ,the more obvious the effect was(P<0 .05) .The ratios of CD4+ ,CD8+ T cells and the frequen‐cies of CD4+CD25+ Treg after PCI for 1 month and 6 months in high‐load atorvastatin showed significant difference compared with those in ordinary‐load atorvastatin group and routine group(P<0 .05) .Conclusion Atorvastatin could regulate the balance of T cell subsets in patients with UA ,and thus it may reduce the UA onset and the treatment effect of high‐load atorvastatin is more sig‐nificant .

ObjectiveTo investigate the method of establishing damaged endometrial stromal cells (ESC) model in vitro.Methods ( 1 ) From June to December 2011 ESC from normal endometrim at proliferation phase ( n =8 ) and secretory phase ( n =8 ) were isolated,cultured and identified in vitro.( 2 ) ESC was treated with different concentrations of mifepristone or withdrawal of mifepristone at different time point.The proliferation inhibition percent was measured by cell counting kit-8 ( CCK-8 ). ( 3 ) 0 μmol/L (control group)and 60 μmol/L(experimental group) concentration of mifepristone was added into ESC for 48 hours,then withdrew of mifepristone,continued to be cultured for 48 hours.The morphological changes were observed and apoptosis of ESC in different menstrual cycle were detected by flow cytometry.The mRNA and protein level of vascular endothelial growth factor ( VEGF),caspase-3,8,and 9 were determined by one-step quantitative real-time PCR (Q-PCR) and western blot.Results( 1 ) ESC from 16 specimens of endometrium were all isolated and cultured successfully. (2) The proliferation inhibition rate of ESC was correlated with concentration and duration of mifepristone positively. The proliferation of ESC could be recovered at a range of time after withdrawal of mifepristone.However,when the concentration of mifepristone was 100 μmol/L,the growth of ESC recovered very hardly. (3) The damaged ESC spacing increased,the spindle shape and vacuolization in the cytoplasm were observed in experimental group; the rate of apoptosis of these damaged cells was significantly increased compared with control groups,which were (52 ± 12)％ vs.( 13 ± 5 ) ％ at the proliferative phase and (53 ± 6) ％ vs.( 32 ± 3 ) ％ at the secretory phase ( all P ＜0.05).The relative mRNA level of VEGF was 0.52± 0.12 in experimental group and 1.00 ± 0.17 in control group at proliferation phase (P ＜0.05).And the relative mRNA level of VEGF was 0.19 ±0.03 in experimental group and 0.81 ±0.07 in control group at secretory phase (P ＜ 0.05).The relative level of VEGF protein in the experimental group were both decreased 1.98 and 2.79 folds at the proliferation phase and the secretory phase when compared with those in control group,respectively ( P ＜ 0.05 ).While the relative levels of caspase-3,8,9 mRNA were 5.62 ± 0.65,5.41 ± 0.53,7.22 ± 0.51 in the experimental group and 1.00 ± 0.44,1.00 ± 0.21,1.00 ± 0.32 in control group at the proliferative phase.In the mean time,the relative levels of caspase-3,8,9 mRNA were 10.22 ± 0.72,25.3 ± 1.72,9.48 ± 1.89 in experimental group and 1.42 ± 0.14,1.14 ± 0.28,1.16 ± 0.12 in control group at the secretory phase,respectively (P ＜ 0.05).Compared with the control group,the levels of caspase protein in the experimental group were increased 2.04 and 1.60 folds in caspase-3,4.23 and 1.49 folds in caspase-8,2.65 and 3.5 folds in caspase-9 at the proliferative phase and at the secretory phase,respectively (P ＜ 0.05 ).ConclusionThe damaged model of ESC can be established after 48 hours by the withdrawal of 60 μmol/L mifepristone in treatment of ESC for 48 hours.

AIM: To investigate the effects of angiotensin-Ⅱ (Ang-Ⅱ) on the proliferation and collagen synthesis in rat cardiac fibroblasts and the expression of Ang-Ⅱ receptor on fibroblasts. METHODS: [3H]-TdR and [3H]-prolin at different concentrations were incubated with the confluent fibroblasts of newborn rats stimulated by Ang-Ⅱ. Receptor of Ang-Ⅱ on fibroblasts and intracellular free calcium ions were examined by autoradiograms and fluorescence technique. RESULTS: In the presence of Ang-Ⅱstimulation, the increase in incorporation of [3H]-TdR and [3H]-prolin was much higher than that of control in a dose-dependent manner. Autoradiograms showed that Ang-Ⅱ was uniformly distributed over the membrane of the fibroblasts. The silver grains on fibroblasts were obviously decreased after adding unlabeled Ang-Ⅱ by the autoradiograms. The concentration of free calcium ions was increased in fibroblast. CONCLUSION: These findings suggest that Ang-Ⅱ promotes fibroblast proliferation and the syntheses of collagen protein, in which Ang-Ⅱ binds to the membrane receptor of fibroblasts to activate the signal system in cells, such as intracellular free calcium ions. [

A new method for O-ethyl S-[2-( diisopropylamino) ethyl] methylphosphonothiolate ( VX) , sarin detection and its kinetic analysis based on piezoresistive microcantilever aptasensor was developed, where VX, sarin aptamers were immobilized on the microcantilever surface by biotin-avidin binding system. A linear relationship between the response voltage and the concentration of VX in the range of 2-60μg/L was obtained. The linear regression equation was △Ue=0. 886C-1. 039 (n=5, R=0. 984, p<0. 001) and the detection limit was 2μg/L ( S/N≥3 ) . A linear relationship between the response voltage and the concentration of sarin in the range of 10-60 μg/L was obtained, the linear regression equation was △Ue=0. 716C-2. 304 ( n=5, R=0 . 996 , p<0 . 001 ) and the detection limit was 10 μg/L ( S/N≥3 ) . The sensor showed no response for O-butyl methylphosphonochloridate, a structural analog of VX and sarin, which indicated high specificity and good anti-interference ability. On this basis, a reaction kinetic model based on receptor-ligand binding and the relationship with output voltage change was established. Response voltage (△Ue ) and response time( t0 ) were obtained from the fitting equation on different concentrations of VX, sarin fitted well with the measured values.

A new magnetic affinity immunoassay (MAIA) strategy based on enzyme-labeled phage displayed antibody was developed. The assay consisted of a sandwich format in which immobilized polyclonal antibody (pcAb) on magnetic microparticle was used for capture probe, and enzyme-labeled phage displayed antibody for specific detection probe to increase enzyme amount and enhance detection signal. By the proposed method,β-bungarotoxin (β-BGT) was successfully detected. A linear relationship between absorbance value and the concentration of β-BGT in the range of 0. 016-62. 5 μg / L was obtained. The linear regression equation was Y=0. 641X+1. 355 (R =0. 9925, n = 13, p<0. 0001) with a detection limit of 0. 016 μg / L. In comparison with the traditional ELISA, this method gave a 10-fold better sensitivity in β-BGT detection. This strategy also gave a 4-fold better sensitivity comparing with the MAIA based on enzyme labeled monoclonal antibody (mcAb). Due to low detection limit, acceptable reproducibility and high specificity, this method holds great promise in toxin trace detection.

This study assessed the effects of leukemia-related protein 16 (LRP16) on the regulation of pancreatic functions in mouse insulinoma (MIN6) cells. Cells with down-regulated expression of LRP16 were obtained by a shRNA interference strategy. Insulin content and glucose-stimulated insulin secretion (GSIS) were examined by radioimmunoassay. Western blotting was applied to detect protein expression. Glucose-stimulated sub-cellular localization of PDX-1 was immunocytochemically determined. Cell proliferation and apoptosis were detected by flow cytometry. Our results showed that LRP16 regulated insulin content in MIN6 cells by controlling expression of insulin and insulin transcription factors. LRP16 gene silence in MIN6 cells led to reduced cell proliferation and increased apoptosis. The observation of phosphorylation of serine-473 Akt and the localization of PDX-1 to the nucleus under glucose-stimulation exhibited that LRP16 was a component mediating Akt signaling in MIN6 cells. These results suggest that LRP16 plays a key role in maintaining pancreatic β-cell functions and may help us to understand the protective effects of estrogen on the functions of pancreatic β-cells.

Objective To investigate the renal expression changes of microRNA-215(miR-215) and its role in diabetic nephmpathy of type 2 diabetic db/db mice. Methods Fourweek-old diabetic db/db mice and norml control group non-diabetic db/m mice were selected.Real-time PCR was used to detect the relative level of miR-215 at the age of 8,12 and 16 weeks.Catenin beta interacting protein 1 (CTNNBIP1) mRNA and protein level were measured by realtime PCR,WesteRN blotting and immunohistochemisty.A lueiferase reporter assay was used to determine whether CTNNBIP1 was a direct target of miR-215. Results (1)With the growth of db/db mice,the major pathological characteristics of kidney included glomerular hypertrophy,segmental mesangial cells proliferation and mesangial matrix expansion.(2)Compared with the db/m mice,the db/db mice of 8,12 and 16 weeks showed obvious increase in body weight(BW),blood glucose (Glu) and 24 hour urinary albumin excretion (UAE) (P＜0.05,respectively).(3)Compared with the db/m mice,special miR-215 was highly expressed in the kidney of db/db mice and was up-regulated significantly according to the development of DN (P＜0.05).(4)The mRNA and protein expression of CTNNBIPl of kidney were consistently down-regulated in db/db mice than those in controls (P＜0.05,respectively). (5)By luciferase reporter,miR-215 could negatively regulate CTNNBIP1 gene by targeting its 3'-UTR sequence (P＜0.01). Conclusion High expression level of miR-215 plays a potential role in the initiation and progression of DN by down-regulating the expression of CTNNBIPl.

Clinical characteristics,including primary and secondary sexual characteristics,basal endocrine profiles,and imaging results were reviewed.Follow-up data were recorded.16 patients had normal karyotypes,manifest amastia,infantile genitalia,amenorrhea,and delayed epiphyseal fusion at the knee and wrist joints.Serum gonadotropic hormone levels were significantly below normal values.15 patients underwent a gonadotropin-releasing hormone (GnRH) stimulation test and 6 patients had a prolonged GnRH stimulation test.16 patients underwent pituitary or brain magnetic resonance imaging (MRI),which showed small pituitaries in three patients,wing tips of suspicious nodules in 2 patients,an empty sella turcica in 1 patient,and a missing right olfactory bulb and tract in 1 patient.1 patient had no detectable uterus or accessory organs,while the other patients had primordial uteri.1 patient was diagnosed as a case of severe osteoporosis.1 patient suffered from pituitary stalk interruption syndrome.An artificial menstrual cycle due to hormone replacement therapy was not sustained after discontinuation of hormone therapy.As disease severity and the date of initiating hormone replacement varied,the results of treatment were quite different.For patients of reproductive age,it was rare to see a reversal of idiopathic hypogonadotropic hypogonadism after discontinuation of hormone therapy.

To analyze the reasons of impaired vision after LASlK and explore its preventive treatment measures preliminarily.METHODS: ln this retrospective study, 175 eyes of 134 patients whose vision was decreased after LASlK were included. The constituent ratio of every reason was counted and uncorrected visual acuity ( UCVA ) between pre-treatment and post-treatment were compared by paired t-test respectively.RESULTS:The overall incidence of impaired vision after LASlK was 1. 86%. The constituent ratio of regression was 51. 43% and UCVA increased from 0. 61±0. 22 to 0. 90±0. 38 (t=8. 00, P<0. 001) after treatment. The constituent ratio of punctate corneal epithelial defect was 32. 57% and UCVA increased from 0. 60±0. 19 to 1. 20±0. 24 (t=20. 00, P<0. 001 ) after treatment. The constituent ratio of accommodative spasm was 5. 14% and UCVA increased from 0.76±0. 21 to 1. 32±0. 22 (t=8. 14, P<0. 001) after treatment. The constituent ratio of corneal flap shift and gauffer was 4% and UCVA increased from 0. 29 ± 0. 26 to 1. 24 ± 0. 28 ( t = 6. 33, P<0. 001 ) after treatment. The constituent ratio of corticosteroid - induced ocular hypertension was 4% and UCVA increased from 0. 57±0. 05 to 1. 0 ± 0. 16 ( t= 2. 53, P<0. 05 ) after treatment. The constituent ratio of fundus lesions and diffuse lamellar keratitis ( DLK) was 2. 86% and UCVA all increased by different degrees after treatment.CONCLUSlON: The reasons of impaired vision after LASlK are many and varied. These cases could recover their vision by discovery and treatment in time, and the appropriate preventive measures were essential.