Rhynchosia volubilis Lour has been a major drug in a folk prescription for contraception in China, whereas its mechanism remains unknown. Its antifertility effects on male mice and antimicrobial activities on sexually transmitted infection (STI) pathogens were previously reported. This study was undertaken to develop the n-Butanol extract of Rhynchosia volubilis Lour (BERVL) as a spermicidal agent with STI prevention. The spermicidal activities of BERVL with different doses were assessed using selected high-motile sperms of normal human semen samples, and their inhibitory effects on Lactobacillus acidophilus were determined. The mechanism of the spermicidal activity was explored by aqueous Eosin Y and Hoechst 33342/PI staining. The results showed spermicidal activities and inhibitory effects of BERVL on Lactobacillus acidophilus were dose-dependent. Dose of 90 mg/mL BERVL terminated all progressive sperm motility within 2 min, and had slight inhibitory effect on Lactobacillus acidophilus, suggesting it was an effective and safe dose for contraception use. About 80% sperms exposed to BERVL displayed changes consistent with high permeability of head membrane. It is concluded that BERVL as spermicide has advantages over N-9 with strong ability to instantaneously kill human sperm and possesses light inhibitory effect on Lactobacillus acidophilus.

Objective To observe the influence of recombinant human thyrotropin(rhTSH)on serum concentration of endogenous thyrotropin(TSH), free triiodothyronine(FT3), free thyroxine(FT4), thyroglobulin antibody(TGAb), and thyroglobulin(Tg). To evaluate the efficacy of rhTSH-aided radioiodine treatment in patients with differentiated thyroid carcinoma(DTC). Methods The study recruitment took place between November 2007 and March 2009. 62 patients(including 45 females)with biopsy confirmed DTC had undergone total or nearly total thyroidectomy, and received 131I treatment. 31 patients(including 22 females), median age of 45 years(23-72), received radioiodine treatment 4 weeks after L-thyroxine(T4)withdrawal. The other 31 patients(including 23 females), median age of 44 years(14-70), underwent rhTSH-aided radioiodine treatment. Before and after rhTSH injection, serum TSH, FT3, FT4, TGAb, and thyroglobulin were tested. Post-radiotherapy whole body scan was performed 5 to 7 days after radioiodine treatment and qualitatively and blindly evaluated by two nuclear medicine physicians. Follow-up took place 6 to 12 months after radioiodine treatment. The efficacy of rhTSH-aided radioiodine treatment was evaluated by whole body scan with diagnostic dose radioiodine. SPSS 13.0 statistical software was applied. Results (1)Before and after rhTSH-aided radioiodine treatment, the serum TSH was(1.08±4.01)vs(140.26±27.20)mIU/L(P＜0.05), thyroglobulin(23.75±132.92)vs(169.58±178.49)μg/L(P＜0.05), FT3(4.52±1.16)vs(4.42±1.11)pmol/L(P＞0.05), and FT4(15.09±5.83)vs(13.66±5.85)pmol/L(P＞0.05),respectively.(2)rhTSH-aided radioiodine ablation treatment had the same effect as L-T4withdrawal aided. The complete response ratio was 77.4% vs 71.0%(P＞0.05)by radioiodine whole body scan of diagnostic dose. Conclusion rhTSH-aided radioiodine treatment of DTC was effective and safe, and did at least at equivalent degree as did L-T4withdrawal. Furthermore, Serum thyroglobulin level could be effectively stimulated by rhTSH with tumor relapse or metastasis.

Objective To investigate whether the release of fibroblast growth factor-1 ( FGF-1 ) was changed after inhibition of S100A13 gene (small hairpin RNA, shRNA)and serum-deprivation in human thyroid cancer cells (TT cells ). Methods The S100A13-shRNA pENTRTM/U6 entry vector was transfected into TT cells. The expression of S100A13 mRNA and protein was detected by immunoflurescence, real-time RT-PCR, and Western blot. Then TT cells were treated with S100A13 gene inhibition and serum-deprivation. The changes in release of FGF-1 were detected by indirect immunoflurescence, RT-PCR, and ELISA. Results S100A13 shRNA transfected TT cells (S100A13 RNAi cells)had a reduction of S100A13 gene and protein expression by 80%.Indirect immunofluorescence indicated FGF-1 was mostly localized in the cytoplasm and nucleus of TT cells in primary culture. When serum-deprivation stress was given to TT cells, FGF-1 in cytoplasm almost disappeared in the cells at 6 h. RT-PCR indicated that when serum-deprivation stress was given to TT cells the mRNA of FGF-1 was reduced. ELISA showed that with inhibition of S100A13, the release of FGF-1 was reduced (P＜0.05).Conclusion S100A13-shRNA pENTRTM/U6 entry vector transfected TT cells may inhibit the expression of S100A13 and reduce the release of FGF-1.

OBJECTIVETo explore action mechanism of electroacupuncture (EA) at "Fenglong" (ST 40) for treatment of hyperlipidemia.

METHODSForty SPF-grade SD rats were randomly divided into a normal group (group A), a model group (group B), a model diet-control group (group C), a model + EA group (group D) and model diet-control + EA group (group E), 8 cases in each one. The rats in group A were fed with normal diet continuously while those in the rest 4 groups were fed with high-fat diet to establish hyperlipidemia model. Afterwards, the rats in group C and group E were fed with normal diet, while EA at "Fenglong" (ST 40) was applied in group D and group E, 30 min per time, once a day. After 28 days of treatment, macrophage was collected in each group. Oil red O-staining was applied to detect the formation of foam cells, and enzyme-linked immunosorbent assay (ELISE) was adopted to measure the contents of total cholesterol (TC) in macrophage and analyze the rate of cholesterol efflux.

RESULTSThe counts of positive cells of oil red O-staining and the contents of TC in the group B, group C and group D were significantly increased compared with those in the group A (all P < 0.01). The counts of positive cells and contents of TC in the group C, group D and group E were significantly clecreased compared with those in the group B (all P < 0.01), and the decline in group D was more obvious than that in the group C (all P < 0.01). Compared with group C and group D, the counts of positive cells and contents of TC in the group E was obviously decreased (all P < 0.01), which was not statistically different from group A (P > 0.05). In the meantime, compared with group C, the rate of cholesterol efflux in group D and group E was significantly increased (both P < 0.01), and the rise in group E was more obvious than that in the group D.

CONCLUSIONThe electroacupuncture at "Fenglong" (ST 40) could significantly prohibit the transformation of macrophage into foam cell and increase rate of cholesterol efflux in macrophage, which could prevent and reverse the formation of foam cell and play an essential role in treating hyperlipidemia and stopping it from developing into a further level.

Acupuncture Points ; Animals ; Biological Transport ; Cells, Cultured ; Cholesterol ; metabolism ; Electroacupuncture ; Foam Cells ; metabolism ; Humans ; Hyperlipidemias ; metabolism ; therapy ; Macrophages ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

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

OBJECTIVETo study the effects of drug plasma of musk and olibanum (DP-M&O) on the release of inflammatory cytokines from monocytes and the expressions of the proteins associated with inflammation of prostatic or endothelial cells induced by prostate antigen (PAg) stimulation.

METHODSWe prepared DP-M&O using SD rats and monocytes and PAgs using BALB/c mice. We pre-treated the monocytes with DP-M&O at the gradient concentrations of 0, 2.5, 5, 10, and 20% for 1 hour, activated them with PAgs, and then cultured them for 96 hours, followed by detection of the release of inflammatory cytokines. We co-cultured the prostate RWPE-1 cells with the endothelial EA. hy926 cells, pre-treated them with the same gradient concentrations of DP-M&O as above for 1 hour, activated with PAgs, and cultured for 96 hours. Then we determined the expression levels of the proteins associated with inflammation of RWPE-1 and EA. hy926 cells by Western blot.

RESULTSDP-M&O decreased the levels of TNF-alpha, IL-1beta, IL-6, and IL-8 and increased that of IL-10 in a concentration-dependent manner. Significant differences were found between the 20% P-M&O and PAg groups in the release of the inflammatory cytokines TNF-alpha (70.8 +/- 22.3 vs. 277.1 +/- 65.5, P < 0.01) , IL-113 (277.5 +/- 22.6 vs. 630.4 +/- 89.7, P <0.01), IL-6 (232.7 +/- 62.7 vs. 994.2 vs. 182.3, P < 0.01), IL-8 (227.3 +/- 79.2 vs. 769.3 +/- 284.1, P < 0.01), and IL-10 (640.2 +/- 201.2 vs. 271.1 +/- 55.8, P < 0.01). Compared with the PAg group, the 10 and 20% P-M&O groups showed remarkable decreases in the protein expression of MCP-1/CCL2 in the RWPE-1 cells (1.12 +/- 0.34 vs. 0.56 +/- 0.11 and 0.34 +/- 0.08) and that of VCAM-1 in the EA. hy926 cells (0.94 +/- 0.22 vs. 0.52 +/- 0.17 and 0.38 +/- 0.12) (P < 0.05 or 0.01).

CONCLUSIONThe compatibility of musk and olibanum can decrease the expression of MCP-1/CCL2 in prostate cells and VCAM-1 in vascular endothelial cells, blocking the adhesion of leucocytes and suppressing inflammatory response.

Animals ; Blotting, Western ; Cytokines ; metabolism ; Endothelial Cells ; drug effects ; metabolism ; Fatty Acids, Monounsaturated ; pharmacology ; Frankincense ; pharmacology ; Inflammation ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-1beta ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; Male ; Mice ; Mice, Inbred BALB C ; Monocytes ; drug effects ; metabolism ; Prostate ; cytology ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo investigate the effect of Bushengzhuyang Fang (Yangjing Capsule, YJC) on penile erectile function and its action mechanisms in rats.

METHODSFifty-six male SD rats were randomly divided into seven groups of equal number: blank control, daidzein, daidzein + testosterone, daidzein + sildenafil, daidzein + low-dose YJC, daidzein + medium-dose YJC, and daidzein + high-dose YJC. The rats in the blank control group were treated intragastrically with normal saline and those in the other groups with daidzein at the dose of 100 mg per kg per day for 30 days. Then the last five groups received additionally testosterone (4 mg per kg per day), sildenafil (2.5 mg per kg per day), low-dose YJC, (0.315 mg per kg per day), medium-dose YJC (0.63 mg per kg per day), and high-dose YJC (1. 26 mg per kg per day), respectively. At 0, 30 and 60 days of treatment, we observed the apomorphine-induced spontaneous erectile response and pathological changes in the corpus cavernosum of the rats, recorded the number of penile erection and erectile incubation period, and determined the serum levels of testosterone (T) and luteinizing hormone (LH).

RESULTSAt 30 days of treatment, the number of apomorphine-induced erections was decreased, the erectile incubation period prolonged, and the serum levels of T and LH reduced remarkably in all groups of rats (P < 0.05). Compared with the findings at 30 days, the number of penile erections was significantly decreased at 60 days in the daidzein group (1.39 ± 0.42 vs 2.67 ± 0.33, P < 0.05) and daidzein + low-dose YJC group (1.33 ± 0.49 vs 2.83 ± 0.61, P < 0.05); the erectile incubation period was markedly ex- tended ([16.33 ± 3.11] vs [8.50 ± 0.93] min and [15.50 ± 3.21] vs [8.63 ± 1.54] min, P < 0.05); and the serum levels of T ([5.34 ± 0.89] vs [1.24 ± 0.30] ng/ml and [5.28 ± 1.12] vs [2.07 ± 0.76] ng/ml, P < 0.05) and LH ([3.62 ± 0.37] vs [2.09 ± 0.12] ng/ml and [3.79 ± 0.28] vs [2.17 ± 0.33] ng/ml, P < 0.05) were significantly reduced in the daidzein and daidzein + low-dose YJC groups, respectively. Pathological examination revealed slightly decreased cavernous sinuses and blood vessels in the corpus cavernosum of the rats in the daidzein + testosterone, daidzein + sildenafil, daidzein + medium-dose YJC, and daidzein + high-dose YJC groups as compared with those in the blank control group.

CONCLUSIONHigh-dose Yangjing Capsule is efficacious for the recovery of erectile function in rats, especially for phytoestrogen-induced erectile dysfunction.

Animals ; Apomorphine ; pharmacology ; Drugs, Chinese Herbal ; therapeutic use ; Erectile Dysfunction ; chemically induced ; drug therapy ; Humans ; Isoflavones ; pharmacology ; Luteinizing Hormone ; Male ; Penile Erection ; drug effects ; physiology ; Penis ; drug effects ; pathology ; Phytoestrogens ; Phytotherapy ; Piperazines ; therapeutic use ; Purines ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Sildenafil Citrate ; Sulfonamides ; therapeutic use ; Testosterone ; therapeutic use ; Vasodilator Agents ; therapeutic use

OBJECTIVETo investigate the biological characters of human skin fibroblasts in fibroblast populated collagen lattice (FPCL).

METHODSThe human fibroblasts were cultured in 3D and the collagen of the rat tail was also prepared. They were examined with the comprising cell cycle and apoptosis, mRNA expression of TGF beta1, and fibronectin, and cell morphology.

RESULTSThe flow cytometry showed that the G0/G1, stage cells were 79% +/- 3%, 87% +/- 2% after the 7 days and 14 days separately, and there were not apoptosis peak observed. RT-PCR analysis revealed that the mRNA expression of TGF beta1, and fibronectin had no difference between human skin fibroblasts cultured in 3D and 2D. Electron microscope showed the cells were plenty of chromatin and organelles.

CONCLUSIONSThe proliferation of the human skin fibroblasts in FPCL is slow, but its biological viability is better.

Animals ; Cell Culture Techniques ; Cell Division ; Cells, Cultured ; Collagen ; Extracellular Matrix ; Fibroblasts ; cytology ; Humans ; Rats ; Skin ; cytology ; Tissue Engineering ; methods

Objective To evaluate the clinical application of stress/rest MPI in the patients with 50％-75％ coronary artery stenosis.Methods The criteria for patient selection were that the patients should have at least one main coronary artery with stenosis more than 50％,and the maximal stenosis should be less than 75％ according to CAG.The stress/rest MPI was performed in 2 weeks before or after CAG.A total of 244 patients (178 males,66 females) with mean age (57 ± 10) years were included in this study.Symptom restriction stress test was used and stress MPI was performed 1 - 1.5 h after 99Tcm-MIBI (925 MBq)injection at the exercise peak.Rest MPI was performed within 48 - 72 h after stress MPI.Myocardial ischemia was diagnosed when there was a reduced uptake or even a defect in 2 different tomographic sections or in the same part of a myocardium in the continuous 2 slices.When there was an irreversible reduced uptake or defect,myocardial infarction was given as the final diagnosis.No reduced uptake or defect in all slices was shown as normal.The impact of MPI images on the selection for optimal clinical therapy plans was also discussed.X2 test was used for statistical analysis.Results A total of 340 coronary arteries with stenosis 50％ - 75％ were found by CAG.According to stress/rest MPI results,207 patients (84.8％)presented normal,33 had myocardial ischemia,3 had myocardial infarction,and 1 had both myocardial infarction and ischemia.In abnormal MPI images,there were 61 ischemic segments and 9 infarct segments,which were associated with 43 stenotic arteries (23 LAD,10 LCX,and 10 RCA).Patients were divided into 2 groups according to the results of MPI:Group 1 with normal MPI (207/244,84.8％ ) and Group 2 with abnormal MPI (37/244,15.2％ ).In Group 1,9 patients underwent coronary artery revascularization (PTCA or CABG),and the others had medical treatment.Eight patients had PTCA and 29 patients had medical treatment in Group 2.There was a statistically significant difference between the two groups ( X 2 =11.9,P =0.001 ).Conclusion Stress-rest MPI may be an effective method to evaluate ischemia degree for patients with 50％ -75％ coronary artery stenosis and a useful indicator for the individual treatment.

Objective:To study the characteristics of neonatal fungal sepsis and the difference between bacterial sepsis and fungal sepsis.To improve the understanding of neonatal fungal sepsis.Methods:Clinical data of neonatal fungal sepsis in neonatal intensive care unit (NICU) were collected from 2011 to 2016 in Peking University first Hospital.The clinical characteristics were analyzed retrospectively.The difference between neonatal fungal sepsis and bacterial sepsis was also analyzed.Results:Fifteen cases of neonatal fungal sepsis were recruited.Over the study period,the incidence of neonatal fungal sepsis was 0.52％,while it was 2.5％ in very low birth weight infants.Clinical characteristics were nonspecific.All the infants were treated with parenteral nutrition and broad spectrum antibiotics.Peripheral inserted central catheter (PICC) was placed in thirteen patients.Pathogenic analyses indicated Candida glabrata was the main pathogen in our study.All the pathogens were sensitive to amphotericin B.Only one Candida glabrata was resistant to fluconazole.Thirty-four cases of bacterial sepsis were included.The clinical characteristics and laboratory examination results were compared.The platelet count was 61 × 109/L in fungal group,while the platelet count was 178 × 109/L in bacterial group.There was statistical difference between the fungal group and bacterial group (P =0.004).The rate of thrombocytopenia was 80.0％ in fungal group,while it was 29.4％ in bacterial group.It was much higher in fungal group than in bacterial group (P =0.001).The rate of PICC placement was 86.7％ in fungal group,while it was 55.7％ in bacterial group.It was much higher in fungal group than in bacterial group (P =0.037).Receiver operating characteristic (ROC) curve analysis showed that the cut-off value of the platelet count for the diagnosis of neonatal fungal sepsis was 145 × 109/L (sensitivity 61.8％,specificity 92.9％).All the patients were cured after standardized antifungal therapy.The indicators of liver and renal function were also measured before and after antifungal therapy.No significant difference was observed before and after treatment.Conclusion:The clinical characteristics of neonatal fungal sepsis was nonspecific.Candida glabrata was the main pathogen in our NICU.It can be cured as the result of standardized treatment.Decreased platelet count and PICC placement may indicate the possibility of fungal sepsis in neonates.