Objective To observe the expression of osteoprotegerin (OPG), receptor activator of nuclear factor κβ ligand(RANKL) and receptor activator of nuclear factor κβ (RANK) in bone tissue of rats with chronic fluorosis and to explore the relation between OPG/RANKL/RANK system and bone damage in chronic fluoride poisoning rat and the antagonism effects of Danlan Xianpeng capsule. Methods SD rats were randomly divided into six groups according to body weight (equal male and female in each group): fluorosis group, high dose drug group, medium dose drug group, low dose drug group, control group, borax group(positive control), 12 rats in each group. The control group drank tap water and the remaining 5 experimental groups consumed 50 mg/L fluoride water, and high, medium and low doses drug group took Danlan Xianpeng capsule at doses of 0.8,0.4,0.2 g/kg,borax group took borax at dose of 0.8 g/kg. OPG, RANKL, RANK protein in rat tibial metaphysis was detected by immunohistochemistry at the 6 month. Results Compared with the control group(173.79 ± 5.23, 174.17 ± 5.01,155.63 ± 7.11), the expressions of OPG, RANKL were increased and the expression of RANK was decreased in fluorosis group(156.83 ± 5.80, 157.74 ± 6.70, 173.92 ± 4.37), the difference was statistically significant (all P ＜ 0.05). Compared with the fluorosis group, the expression of OPG and RANKL were decreased and the expression of RANK was increased in high-dose drug group, middle-dose drug group(169.67±5.07, 168.08 ± 5.05,162.12 ± 4.24, 170.78 ± 5.01, 168.41 ± 7.19, 166.69 ± 5.78, all P ＜ 0.05). Compared with the borax group (167.27 ± 4.08, 167.85 ± 5.01, 166.14 ± 3.95), the expression of OPG and RANKL was increased in the low-dose drug group (163.40 ± 4.11, 159.49 ± 5.78), the expression of RANK was increased in the high-dose drug group (162.12 ± 4.24) and decreased in the low-dose drug group(171.54 ± 8.06), the difference was statistically significant (all P ＜ 0.05). Conclusions Chronic fluoride poisoning can cause increased bone turnover and enhance the activity of osteoelastic absorption by increasing RANKL. Danlan Xianpeng capsule can affect bone remodeling through the OPG/RANKL/RANK system, and antagonises bone damage caused by fluoride.

Objective To explore the effect of experimental design in the experimental teaching of fundamental nursing. Methods 168 and 199 baccalaureate nursing students were in the control and experimental group respectively, the control group received traditional experimental teaching method, and the experimental group must complete the experimental design before entering the lab. The teaching effect between the two groups were compared. Results The mean operation score of the experimental group was significantly higher than the control group, the proportion of those students who reflected their teaching method could enhance creative ability, problem-solving ability and practical ability was higher in the experimental group, 100% teachers reflected experimental design could improve operation level, strengthen theoretical knowledge, animate teaching atmosphere. Conclusions Experimental design could arouse students' enthusiasm and initiative to study, advocate them to think and ask.

Objective To investigate the effect of GATA-6 on endogenous carbon monoxide(CO) inhibited pulmonary vascular smooth muscle cells(PVSMCs) proliferation induced by platelet-derived growth factor(PDGF).Methods Tissue mass culture was done to get PVSMCs artery in SD rats.The PVSMCs were stimulated to proliferation by PDGF(20 ?g/L),and the 3 different concentrations of hemin[a substrate and inducer of heme oxygenase-1(HO-1)] add into the cultures to induce CO production.The PVSMC cell proli-feration was detected by 3-(4,5-dimethyl-2-thiazoly1)-2,5-diphenyl-2H-tetrazolium bromide(MTT) method,DNA synthesis was detected by thymidine incorporation,and GATA-6 mRNA expression was detected by reverse transcription polymerase chain reaction(RT-PCR).Results CO inhibited the PVSMCs proliferation induced by PDGF in a dose-dependent and time-dependent manner.Hemin with high concentration markedly inhibited the proliferation and DNA synthesis of PVSMCs.After 2 hours with PDGF,the expression of(GATA-6) mRNA markedly down-regulated,and began returned after 6 hours.However,CO could reversed this down-regulation.Conclusions CO can inhibit the proliferation of PVSMCs induced by PDGF.PDGF can result in the expression of GATA-6 mRNA down-regulated;the down-regulation is reversed by CO.This study suggested that CO maybe inhibit PVSMCs proliferation by regulating expression of GATA-6.

OBJECTIVETo establish a feasible protocol to provide genetic diagnosis and prenatal diagnosis in Chinese hemophilia patients and their relatives by direct exon sequencing.

METHODSIn our study, genetic diagnosis was performed on 5 unrelated families with informed consent, which included 3 pregnant women who asked for prenatal diagnosis. Umbilical cord blood was obtained from 2 fetuses and amniotic fluid from another fetus. After extraction of the genomic DNA, all of the exons, exon-intron boundaries and their flanks of FⅨ gene were amplified by polymerase chain reaction (PCR). PCR products were detected through direct-sequencing.

RESULTSSequence analysis indicated that the patients and carriers from 5 families have the pathogenic mutations,c.1022G>A (p.R341Q), c.484 C>T (p. R162X), c.1135C>T (p.R379X), c.799C>T (p.H267Y), c.1232G＞T (p.S411I), respectively. Except c.484 C>T (p. R162X), 4 of the 5 mutations were reported firstly in Chinese population. During prenatal diagnosis, one of the fetuses was found to be affected with c.484C>T; p.R162X. The remaining two fetuses were diagnosed as normal, the results of which were later verified by post-birth diagnosis, and factor FⅨ activities in plasma was 52.7% and 76.2%, respectively.

CONCLUSIONIn the quest of strict quality control, exon sequence on FⅨ gene was a rapid and accurate method for genetic diagnosis and prenatal diagnosis of hemophilia B.

Exons ; genetics ; Factor IX ; genetics ; Female ; Hemophilia B ; genetics ; Humans ; Male ; Pedigree ; Prenatal Diagnosis ; methods ; Sequence Analysis, DNA ; methods

AIM: To comparatively analyze the application of laser confocal microscopy and corneal smear in the diagnosis of fungal keratitis.METHODS: Totally 77 patients (77 eyes) diagnosed as fungal keratitis were selected.Laser confocal microscopy and corneal smear examination were performed to observe the characteristics of the images, and the detection rate of fungus were compared between the two methods.RESULTS: Of the 77 patients, 66 eyes (86%) were positive and 11 eyes were negative detected by laser confocal microscopy;51 eyes (66%) were positive and 26 eyes were negative detected by corneal smear examination, the difference was statistically significant compared between two group (P<0.05).CONCLUSION: Laser corneal confocal microscopy is relatively safe and sensitive.If combined the application of laser corneal confocal microscopy and corneal smear examination in clinical, the diagnosis rate of fungal keratitis may be improved.

Objective To discuss the correlation between cervical intraepithelial neoplasia(CIN) and uterine cervix cancer and the combination of TCT and colposcope for the investigation of cervical lesion.Methods 5545 patients were preliminarily screened by cervical fluid basement cell folium smear,in which 307 patients are masculine and 219 are negative,the 526 patients are suspected cervical lesions in clinical symptoms and they further underwent colposcopy and biopsy,the results were analyzed by pathohistology which is golden standard.Results CIN coherent dangerous factors analyzed by single factor Logistic regression analysis are:cervical HPV pollution,age of sexual activity,sexual partners numbers,protection of sexual life, STD history,age and frequency of miscarriage ( P < 0.05 ).The coincidence between TCT,colposeopy and pathologic diagnosis are 49.4%, 83.8% ;the omission diagnostic rate of LSIL,HSIL and CC from TCT are 63.63% ,74.44% ,100% ;the omission diagnostic rate of LSIL,HSIL and CC from colposcopy are 33.37%, 12.22% ,0;the sensitivity of TCT,colposcopy and TCT combine colposcopy for CIN are 61.2% ,84.6% ,94.5% and specificity are 85.4% ,88.2.% ,90.8%.Conclusions (1)The generation of CIN and cervical cancer are correlated with cervical HPV pollution,sexual behavior and various kinds of lower genital tract infection.(2) Fluid basement cell folium smear can promote the property of diagnosis,meet the demands of early stage uterus neck cancer and precancer lesion investigation,are suitable to be routine cheek methods in primary hospital.

Objective To evaluate the role of early cleavage embryo in combination with embryo growth rate and morphology scoring in embryo selection in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. Methods Six hundred and ten IVF/ICSI cycles were randomly assigned to group A(269 cycles) and group B(341 cycles). In group A, transferred embryos were chosen according to embryo growth rate and morphology scoring by 72 h(D3) after fertilization, while early cleavage embryo was added to the selecting system in group B. The pregnancy rate and implantation rate were compared between two groups, and the clinic outcomes were compared between transfers with early cleavage embryos and without early cleavage embryos in group B. Results The pregnancy rate and implantation rate in group B were significantly higher than those in group A (P < 0.05). Transfers with early cleavage embryos also achieved much higher pregnancy rate and implantation rate in group B (P < 0.01). Conclusion Compared with embryo growth rate and morphology scoring, early cleavage embryo in combination with embryo growth rate and morphology scoring can improve the clinical outcomes in IVF/ICSI cycles.

To prepare sagittatoside B with epimedin B Hydrolyzed from cellulase. With the conversion ratio as the index, the effects of pH value, temperature, reaction time, dosage of enzyme and concentration of substrates on the conversion ratio were detected. L9 (3(4)) orthogonal design was adopted to optimize the preparation process. Hydrolyzed products were identified by MS, 1H-NMR, and 13C-NMR. The results showed that the optimum reaction conditions for the enzymatic hydrolysis were that the temperature was 50 degrees C, the reaction medium was pH 5.6 acetic acid-sodium acetate buffer solution, the concentration of substrates was 20 g x L(-1), the mass ratio between enzyme and substrate was 3: 5, and the relative molecular mass of the reaction product was 646.23. NMR data proved that the product was sagittatoside B. The process is simple and reliable under mild reaction conditions, thus suitable for industrial production.
Cellulase ; metabolism ; Drug Compounding ; methods ; Flavonoids ; chemistry ; Hydrogen-Ion Concentration ; Hydrolysis ; Temperature ; Time Factors

OBJECTIVETo observe the myocardial protection of cardioplegic solution with Salvia miltiorrhizae (SM) in extracorporeal circulation of coronary artery bypass graft (CABG) and to investigate the mechanisms of SM.

METHODS30 patients who received CABG under extracorporeal circulation were randomly assigned to two groups, the observation group (15 cases) and the control group (15 cases). Patients in the observation group received the cardioplegic solution with SM and those in the control group received the cardioplegic solution without SM. The indices such as serum SOD activities, MDA contents, LDH, CK-MB, cTnl levels, the rate of heart reskip, activated coagulation time (ACT), the time of assisted respiration, and the days of in-hospital after operation were observed in the two groups pre-operation, post-operation, 6 h and 24 h post-operation, respectively.

RESULTSWhen compared with the control group, MDA contents, LDH, CK-MB, cTnl levels were lower, SOD activities (all P<0.05) and heart re-skip rate (P>0.05) higher in the observation group. There was no statistical significance in the time of assisted respiration, the days of in-hospital, or ACT in the two groups (P>0.05).

CONCLUSIONSThe application of cardioplegic solution with SM in extracorporeal circulation of CABG showed obvious myocardial protection. It had better effects than the cardioplegic solution with no SM.

Aged ; Cardioplegic Solutions ; pharmacology ; therapeutic use ; Coronary Artery Bypass ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Extracorporeal Circulation ; methods ; Female ; Humans ; Male ; Middle Aged ; Myocardium ; metabolism ; Salvia miltiorrhiza

This article firstly established a new efficient method for screening anti-osteoporosis ingredients, which used two-dimensional zebrafish model combined with hyphenated chromatographic techniques to evaluate anti-osteoporosis activities of epimedin A and its metabolite baohuoside I. Adult zebrafish was used for metabolism of epimedin A in 0.5% DMSO, and LC-MS was used for analysis of the metabolite, which was captured by HPLC, and prednisolone-induced osteoporosis model of zebrafish was used to evaluate the anti-osteoporotic activities of trace amounts of epimedin A and baohuoside I. The results indicated that epimedin A and baohuoside I can prevent prednisolone-induced osteoporosis in zebrafish. The developed method in this paper enables the separation, enrichment and analysis of micro-amount metabolite of epimedin A, and anti-osteoporosis activities in vivo of epimedin A and baohuoside I was simple and efficient screening resorting to zebrafish osteoporosis mode. This paper would provide new ideas and methods for a rapid and early discovery of anti-osteoporosis activities of micro-ingredients and its metabolite of traditional Chinese medicine.
Animals ; Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; pharmacology ; Flavonoids ; metabolism ; pharmacology ; Mass Spectrometry ; Osteoporosis ; drug therapy ; Zebrafish