Objective Daidzein solid dispersions were prepared by solid dispersion technology to improve in vitro dissolution rate. Methods Daidzein solid dispersions were prepared by solvent method using polyvinyl pyrrolidone K30 ( PVP K30) as carrier.The in vitro dissolution characteristics of solid dispersions were evaluated,and the properties were detected by IR and XRD. Results The dissolution rates of different mass ratio of daidzein-PVP solid dispersion were significantly improved compared with that of daidzein API.The cumulative dissolution of solid dispersion with mass ratio of 1∶6 within 30 minutes was up to 87.8%,equivalent to six times of API. The in vitro drug release kinetics were fitted mathematically to Korsemeyer-Peppas model. Conclusion Solid dispersion with PVP K30 as carrier could significantly improve dissolution rate of daidzein.

Objective To investigate the location,fine structure of melanocytes in human fetal scalp hair follicles.Methods The scalp with hair follicles was obtained from a dead fetus of 6 months of age,and divided into two parts.One part was embedded in paraffin,tissue sections were prepared with a width of 7 μm and stained with NKI/beteb,monoclonal antibodies to HMB-45,tyrosinase and tyrosinase-related protein 1(TRP1),respectively.The other part with hair follicles was treated with collagenase type Ⅱ 0.1 g/L and trypsin,then,cell suspension was collected and cultured.After 14-day culture,follicle melanocyte cells (FMC)were separated from keratinocytes by differential trypsinization,and fibroblasts were removed with geneticin.Following three times of pure passage,FMC were seeded and fixed on mica for scanning electron microscopy(SEM)and atomic force microscope(AFM)scanning.Results Histopathological examination showed that NKI/beteb positive cells located at the outer root sheath of human hair follicles,and these cells stained negatively for HMB-45,tyrosinase and TRP1 antibodies.However,in the hair bulb,lots of cells expressed HMB-45,tyrosinase and TRP1 antigens.After fibroblasts and keratinocytes were removed,two kinds of melanocytes remained in the culture:one was small in number and showed abundant melanin,which was lost after subsequent passage;the othgr was large in number and had no melanin initially,but proliferated very rapidly.After three passages,almost all the melanocytes were positive for NKI/beteb.As SEM and AFM showed,most cultured melanocytes appeared fusiform with two(rarely three)dendrites,and the cell body was round or oval with a few melanosomes scattered in but no clear secondary branches on the dendrites.Conclusions The melanocytes in outer root sheath of hair follicles from the fetal scalp are presumed as melanocyte stem cells or their progenies.In vitro,these cells proliferate very rapidly during early phases,but the morphology and function of them still remain immature,which is unfavorable for melanosome transport.

ObjectiveTo study the dose and time effects of X-ray radiation on the expression of Pokemon gene and protein in human lung adenocarcinoma cell line A549.MethodsA549 cells was exposed to different doses of X-ray (2,4,6 and 8 Gy),and the expression of Pokemon mRNA and protein of the cells was detected by using Quantitative real-time PCR and western-blotting at 2,4,8,12,24,and 48 h after irradiation. 3-( 4,5-Dimethylthiazol-2-yl )-2,5-diphenyltetrazoliumbromidewasused to detectthe proliferation of A549 cells at 1,2,3,4,and 5 d after 2 Gy X-ray irradiation.The mock treated A549 cells were used as the control.ResultsThe expression of Pokemon mRNA trended to decrease after irradiated with 4,6 and 8 Gy in the earlier period and increased in the later period with statistical difference at the most time points (t =3.40 -154.76,P =0.000 -0.041 ).The expression of Pokemon protein trended to increase and reached the peak at 8 h after irradiated of 2,4,6 and 8 Gy with statistical difference at the most time points ( t =4.18 - 89.64,P =0.000 - 0.039).Compared with the control,the proliferation of A549 cells was significantly inhibited during 3 to 5 d after irradiation of 2 Gy ( t =2.34 - 18.19,P =0.000 -0.040).ConclusionsX-ray irradiation may increase the expression of Pokemon mRNA and protein in A549 cells,which might be correlated with radiation-resistance of A549 cells.

Objective To analyze the nursing interventions of two kinds of percutaneous nephrolithotomy (PCNL),so that we can provide personalized service for patients with PCNL and improved quality of nursing. Methods 140 cases of patients undergoing PCNL were chosen from January 2010 to January 2012.70 of them received standard PCNL,another 70 cases were given tubeless PCNL.The same nursing method was given before the operation,and personalized nursing was given according to difference in operation method.A self-designed questionnaire was conducted for satisfaction degree with nursing service of all patients on discharge. Results Satisfaction degree with psychological care,catheter nursing and complication nursing of tubeless PCNL was higher than standard PCNL. Conclusions Two methods of PCNL have become conventional therapy for upper urinary calculus,and careful nursing is very important,especially for tubeless PCNL.

Objective: To observe the clinical efficacy of combined electroacupuncture and nerve growth factor (NGF) injection at acupoints in the treatment of common fibular nerve paralysis and provide evidences for integrative Chinese & western medicine against diabetic peripheral neuropathy (DPN). Methods: Forty subjects were randomized into two groups and NGF injection; and control group was given herbal suffocation, oral Dibazol and compound vitamin B and Mecobalamin Injection. The clinical symptoms and nerve conduction velocity were observed and compared. Results: The cure rate was higher in treatment group than in control group (P<0.05); after treatment, the nerve conduction velocity was improved in both groups (P<0.01), with a significant improvement in treatment group than in control group (P<0.01). Conclusion: Combined electro-acupuncture and NGF injection at acupoints is quite effective in the treatment of common fibular nerve paralysis.

AIM: To investigate the expression of matrix metalloproteinases(MMPs) in pulmonary arterioles of rats with chronic hypoxia and hypercapnia-induced pulmonary hypertension. METHODS: MMP-2, MMP-9 and MMP-2 mRNA, MMP-9 mRNA were observed in pulmonary arterioles by the techniques of immunohistochemistry and in situ hybridization. RESULTS: ①The mean pulmonary artery pressure (mPAP) and weight ratio of right ventricle to left ventricle and septum (RV/LV+S) of hypoxia-hypercapnia groups were higher than those of normal control group ( P

AIM:To investigate the effect of puerarin on pulmonary vessel collagen metabolism in pulmonary hypertension rats induced by chronic hypoxia and hypercapnia. METHODS: Collagen Ⅰ,Ⅲ and their mRNA were observed in pulmonary arterioles by the technique of immunohistochemistry and in situ hybridization. RESULTS: ① Light microscopy showed media thickness of pulmonary arterioles was much higher in HH(hypoxic-hypercapnia) group than that of NC(normal control) group, and, vessel cavity turned more straiter in HH group than that of NC group.However, the damage of pulmonary arterioles in HP(hypoxic-pueratin) group was much slighter than that of HH group. ② The levels of plasma ET-1 and lung homogenates Hyr were much higher in HH group than those of NC group( P 0.05). CONCLUSION: Puerarin inhibited the deposition of collagen and improved pulmonary vessel remodeling.

Objective: Duchenne and Becker muscular dystrophy (DMD/BMD) is an X-linked lethal recessive disease caused by mutations in the dystrophy gene. There is no efficient treatment for this serious and disabling disease. We established a combination method to detect carriers and perform prenatal diagnosis. Methods: In our study, from 1994 to 2005, using a different combination of 5 methods, including SRY gene amplification, multiplex PCR, multiplex Fluorescence PCR capillary electrophoresis, multiplex ligation-dependent probe amplification (MLPA) and linkage analysis of short tandem repeats (STR), 36 prenatal diagnosis were performed for pregnancies at risk of having a DMD/BMD baby through amniocentesis. Results: Fourteen out of 21 male fetuses were found to be affected and respective pregnancies were terminated. A combined diagnostic rate of 83% was achieved for 30 cases with deletions, duplications, and non-deletion mutations after tested by more than one method. Conclusion: Using a combined method, we can diagnoses patients and carriers in DMD families, and perform prenatal diagnosis for the risk fetus. MLPA provides a simple, rapid and accurate method for deletions and duplications of all the 79 DMD exons. MLPA method for DMD diagnosis is the first report in our country.

Objective To investigate the location and expression of hypoxia inducible factor (HIF) subunits in the remnant kidney of 5/6 nephrectomy rats. Methods Remnant kidneys were produced in adult male SD rats by 5/6 nephrectomy. The renal function and histopathological changes were evaluated at week 1, 2, 4, 6, 8 and 12 after operation. Tissues of remnant kidneys were collected to detect the location and expression of HIF-1α and HIF-2α by immunohistochemistry staining and Western blotting. The mRNA levels of HIF targeted genes vascular endothelial growth factor (VEGF) and heme oxygenase-1 (HO-1) were determined by RTPCR. Results (1) 5/6 nephrectomy rats underwent one week of acute renal failure at first[Scr (122.8±22.1) μmol/L] and then developed compensative chronic renal failure [(66.0±3.7)-(66.4±8.4) μmol/L], but the level of Scr increased quickly after week 6 [(66.4±8.4)-(127.8±22.7) μmol/L],concomitantly with progressive tubulointerstitial fibrosis in remnant kidney cortex. (2) In cortex, HIF-1α was expressed only in the atrophic and dilated tubular cells while HIF-2α was located in endothelial, interstitial fibroblasts, and vascular smooth muscle cells. The semiquantitative results of imunohistochemistry and Western blotting revealed that HIF-1α and HIF-2α were both gradually up-regulated during the early stage of remnant kidney, peaked at week 4 and 6, and then gradually down-regulated. (3) The mRNA levels of HIF targeted genes VEGF and HO-1 transiently peeked at week 4 and 6, and then decreased gradually. Conclusions The increased stabilization of HIF-αprotein and transcription of HIF targeted genes at the early stage of this model is a compensation reaction towards hypoxia. The mechanism of decreased expression of HIF-α at the end stage of chronic kidney disease deserves further investigation.

Objective: To investigate the relationship between blood triglyceride (TG) level and brachial-ankle pulse wave velocity (baPWV) in relevant population. Methods: A total of 22622 participants in Kailuan area were enrolled and divided into 4 ranks: Stroke rank, n=5219, Aged rank n=1754, Pregnancy hypertension Rank, n=1561and Peripheral vascular diseases rank, n=14088. All participants finished baPWV examination. The relationship between TG and baPWV was studied by partial correlation analysis, the impact of TG on baPWV was assessed by multivariate linear regression and multivariate Logistic regression analysis. Results: There were 21713 subjects finally enrolled for cohort study and 909 subjects excluded for missing TG information. With elevated TG levels, the average baPWV and the detection rate of baPWV≥1400 cm/s showed the increasing trend. Partial correlation analysis presented that TG level was positively related to baPWV, r=0.235; with adjusted age and gender, TG was still positively related to baPWV, r=0.080. Logistic regression analysis indicated that with other adjusted confounding factors, compared with the first quartile of TG, the second quartile, third quartile and fourth quartile of TG were the risk factors for baPWV≥1400 cm/s, OR=1.161, 95% CI (1.035-1.302), OR=1.312, 95% (1.165-1.476) and OR=1.652, 95% CI (1.463-1.865) respectively. Conclusion: Blood TG level was positively and independently related to baPWV in relevant population.