Oxidative stress induces apoptosis of many cells and arrest of their differentiation.Both Danshensu(DSS)and hydrogen sulfide(H2S)produce significant antioxidant effect in various systems. In this study,we synthesized SDSS,a new H2S-releasing compound derived from DSS,and studied its antioxidant effect in an H2O2-induced MC3T3-E1 osteoblastic cell injury model. We first characterized the H2S releasing property of SDSS in both in vivo and in vitro models. HPLC chromatogram showed that intravenous injection of SDSS in adult rats released ADT-OH,a well proved H2S sustained-release moiety, within several minutes in the rat plasma. Using an H2S selective fluorescent probe, we further confirmed that SDSS released H2S in MC3T3-E1 osteoblastic cells. Biological studies revealed that SDSS had no significant toxic effect but produced protective effects against H2O2-induced MC3T3-E1cell apoptosis. SDSS also reversed the arrest of cell differentiation caused by H2O2treatment. This was caused by the stimulatory effect of SDSS on bone sialo protein,runt-related transcription factor-2, collagen expression, alkaline phosphatase activity, and bone nodule formation. Further studies revealed that SDSS reversed the reduced superoxide dismutase activity and glutathione content,and the increased ROS production in H2O2treated cells. In addition, SDSS significantly attenuated H2O2-induced activation of p38-, ERK1/2-, and JNK-MAPKs. SDSS also stimulated phosphatidylinositol 3-kinase/Akt signaling pathway.Blockade of this pathway attenuated the cytoprotective effect of SDSS.We also observed the effect of SDSS on aspirin-induced gastric injury and found that SDSS protected against aspirin-induced gastric damage. In conclusion, SDSS protects cells against H2O2-induced apoptosis by suppressing oxidative stress.

Objective To discuss the influence of different concentration sulindac on pancreatic cancer cell line PANC-1 cell proliferation and apoptosis,and investigate the possible mechanism that sulindac can inhibit the Wnt/beta-catenin pathway to kill pancreatic cancer cells. Methods PANC-1 cell were divided into negative control group (added containing no sulindac DMSO)and experimental group (added sulindac with concentrations of 0.25 ,0.5 ,1 ,1.5 ,2 mM medium,respectively,name as 0.25 mM group,0.5 mM group,1.0 mM group,1.5 mM group,2.0 mM group),and treated with different time,cell proliferation inhibition ratio in each group was detected by MTT assay,cell apoptosis ratio was detected by flow cytometry,the expression ofβ-catenin mRNA and protein were detected by RT-PCR and immunocytochemistry.Results MTT results showed that sulindac can inhibit the cell proliferation of PANC-1 by a dose-and time-dependent manner.Cell apoptosis increased after sulindac treatment in different degrees,and there were statistical differences between 1.5,2.0 mMgroup and control groups (P<0.05).RT-PCR results showed that the expression ofβ-catenin mRNA decreased after the treatment of sulindac,there were statistical differences between 1.5,2.0 mMgroup and control group (P<0.05). In the 2.0mM group,the expression ofβ-catenin decreased along with the time extending (P<0.05 ).ICC results showed that sulindac inhibited the expression ofβ-catenin protein and nuclear accumulation,there were no statistical differences in 0.25 ,0.5 mM group and control group,but there were statistical differences in 1.0,1.5,2.0 mMgroup.Conclusion Sulindac could inhibit cell proliferation and facilitate apoptosis of PANC-1,this effect is dose-and time-dependent.The inhibition of Wnt/beta-catenin signal pathway may be a possible mechanism of its cytotoxicity.

Objective:To establish the determination method for icariin in Zhuyun Capsules. Methods: The main active component icariin of Herba Epimedii was determined by TLC scanning. Results: This methods was quick, simple, accurate and reproducible. Conclusion: This method can be used as one of quality control standards for Zhuyun Capsules.

BACKGROUND:Basic fibroblast growth factor(bFGF)can promote production of collagen,fibronectin and matrix enzyme in healing wounds.However,dysregulation of this process,such as the abnormal coordination of cell proliferation,extracellular.matrix and neovasculadzation formation,or remodeling of the wound matrix will lead to excess accumulation of scar tissues.OBJECTIVE:To investigate effects of bFGF on normal skin wound healing and hypertrophic scar formation.METHODS:Normal and hypertrophic scar fibroblasts from tissue biopsies from 5 patients who underwent plastic surgery for repairing hypertrophic scars were isolated and cultured.The expressions of collagen,fibronectin and protein synthesis were detected by RT-PCR and ELISA.The mitochonddal membrane potential changes were measured using JC-1 staining and flow cytometry.Simultaneously,adenosine tdphosphate(ATP)levels were determined by chemiluminescence method.The effects of bFGF on these indexes of normal and hypertrophic scar fibroblasts were observed.RESULTS AND CONCLUSION:Hypertrophic scar fibroblasts become slower after being exposed to bFGF,which selectively inhibited type Ⅰ collagen production in hypertrophic scar fibroblasts(P<0.05).Although bFGF inhibited type]collagen production,it had no effect on type Ⅲ collagen expression in both normal and hypertrophic scar fibroblasts.However,fibronectin expression in the normal fibroblasts was up-reguleted after bFGF treatment(P<0.05).In addition,the mitochonddal membrane potential tended to depolarization,although no statistical difference,in hypertrophic scar fibroblasts treated with bFGF(10 or 100 μg/L).bFGF treatment increased the cellular ATP levels in the normal fibroblasts,while there were no significant alterations in the hypertrophic scar fibroblasts over a treatment of bFGF(10 or 100 μg/L,P<0.05).The results suggest that there are differential effects and mechanisms on the skin fibroblasts with bFGF treatment in normal wound healing and hypertrophic scar formation.

Objective To evaluate the maternal and fetal outcomes of pregnant women with lupus nephritis (LN) and the risk factors.Methods Ninety-three patients with 97 pregnancies from January 1st,1990 to December 31st,2012 in Peking Union Medical College Hospital were evaluated retrospectively.Objects of study were divided into three groups:stable lupus before pregnancy (stable group,52 cases),active lupus before pregnancy (active group,26 cases),and newly diagnosed LN during pregnancy (19 cases).Adverse maternal outcomes included exacerbated disease during pregnancy,preeclampsia,increased proteinuria and impaired renal function during pregnancy or postpartum,maternal death,thrombocytopenia and hypocomplementemia.Adverse fetal or neonatal outcomes included therapeutically termination of pregnancy,fetal loss,neonatal death,preterm labor,small gestational age and asphyxia.Statistical analysis was performed by Chi-square test or Fisher's exact test.A binary logistic regression model was used to evaluate the risk factors for adverse maternal and fetal outcomes.Results (1) Adverse maternal outcomes:There was no significant difference between exacerbated cases during pregnancies in stable group and that in active group [53.8 ％ (28/52) vs 61.5 ％ (16/26),x2 =0.417,P＞0.05].After deleting abortions before 20 weeks of gestation (5 cases in stable group and 4 cases in active group),there was no significant difference between preeclampsia incidence in stable group and that in active group [36.2％ (17/47) vs 59.1％ (13/22),x2 =3.204,P＞0.05].In nineteen newly diagnosed LN women,eighteen cases were over 20 weeks of gestation,during which preeclampsia incidence was 6/18.(2) Adverse fetal or neonatal outcomes:Therapeutically termination of pregnancy rate was higher in active group than that in stable group[42.3％(10/26) vs 7.7％(4/52),Fisher's exact test,P＜0.01].After deleting patients who required termination of pregnancy (three cases in stable group) and therapeutically termination of pregnancy (four cases in stable group and ten cases in active group),the rate of fetal loss and neonatal death was higher in active group than that in stable group [5/16 vs 6.7％(3/45),Fisher's exact test,P＜0.05].The rate of adverse fetal or neonatal outcomes was higher in active group than that in stable group [92.3％(24/26) vs50％(26/52),x2=13.483,P＜0.001].Among the nineteen newly diagnosed LN cases during pregnancy,the numbers of therapeutically termination of pregnancy and fetal loss were five and three cases respectively; among eleven live birth cases,two newborns died from severe asphyxia,and nine cases were preterm birth.(3) Binary logistic regression analysis showed that the independent risk factors for exacerbated lupus during pregnancy were hypocomplementemia (OR =0.300,95％ CI:0.104-0.863) and thrombocytopenia (OR =0.054,95％CI∶0.007-0.439).The independent risk factors for preeclampsia in LN pregnant women were thrombocytopenia (OR=0.151,95％CI:0.046-0.499) and LN recurrence or first diagnosed during pregnancy (OR=0.135,95％CI:0.027-0.679).The independent risk factors for adverse fetal or neonatal outcomes were preeclampsia (OR=0.134,95％CI:0.028-0.637) and lupus active during pregnancy (OR =0.026,95 ％ CI:0.005-0.138).Conclusions Active lupus before pregnancy is associated with poor maternal and fetal outcomes in lupus nephritis pregnancy.All pregnancies with LN should be planned,preferably after more than six months of quiescent disease.Blood pressure,renal function,proteinuria and level of platelet and serum complements should be closely monitored.

Objective To study pharmacokinetics of curcumin in Curcuma phaeocaulis in rats in vivo.Methods HPLC method was used to determine the curcumin in rat plasma.The conditions were column: Lichrospher-5-C_(18)(250 mm?4.6 mm, 5 ?m); column temperature: 25 ℃;mobile phase: CH_3CN-5% HAc water solution(45∶55);flow: 1 mL/min;detection wavelength: 420 nm.Results The calibration curve was liner(r=0.999 5) at the range of 6.5—104 ?g/mL.The average recovery was 98.5%.RSD was 2.41%(n=5).The pharmacokinetic parameters of curcumin were as follows: k_a was 0.53/h,k_e was 0.10/h,t_(1/2ka) was 1.32 h,t_(1/2k) was 6.89 h,t_(peak) was 3.89 h,C_(max) was 93.15 ng/mL,AUC was(1 369.38) ng/mL.Conclusion This method is stable,simple,and reliable,which can be applied for the determination of curcumin in plasma and pharmacokinetic studies.

Objective Present study aimed to research the mechanism of the use of 0.2% brimonidine tartrate eye drops in the treatment of optic nerve crush injury of rat.MethodsAnimal models of optic nerve crush injury were created in 60 SD female rats by clipping the exposed optic nerve at 2 mm in retrobulbar for 6 seconds with 78 grams of reverse forceps.The successful model was identified as Marcus-gun pupil without bleeding of fundus after operation.The animals were randomly assigned to model group and brimonidine treating group,and another 30 normal SD rats were used as the normal control group.The 0.2% brimonidine tartrate eye drops was topically administered at 2 hours before operation and after operation twice per day in the brimonidine treating group.The retinas from 18 rats were isolated after 3,7,21 days for RGCs counting by H&E staining,and the retinal ultrastructure was examined under the transmission electron microscope.The retinas from the other 72 SD rats (including normal,model and brimonidine groups) were prepared for the detection of bcl-2 and bax using immunohistochemistry l,3,5,7,14,21 days after the operation.ResultsNormal and almost normal retina structure was exhibited in rats of the normal group and brimonidine treating group,but disorder of cellular arragement and decrease of retinal thickness were found in the model rats under the optical microscope.The RGCs counting was significantly different among the three groups from 3 days through 21 days after operation with the considerably declination in the model group and brimonidine treating group compared with the normal control group (P＜0.05-0.01).However,that of the brimonidine treating group was obviously increased in comparison with the model group (P＜0.01).The expression of bax in rat retina was obviously reduced (P＜0.01),but the expression of bcl-2 was increased in brimonidine group compared with the model group from 5 through 7 days after operation (P＜0.01).ConclusionThe 0.2% brimonidine tartrate eye drops has a preventive effect on optic nerve crush injury of rat,and its inhibition on apoptosis is one of the mechanisms.

Objective To compare intravascular ultrasound (IVUS) and coronary angiography in measuring the lumen diameter and coronary arteries stenosis rate, to investigate the impact of IVUS in the choice of surgical indications, surgical procedure guidance and effects of operation.Methods The patients who underwent percutaneous coronary intervention(PCI) therapy from may 2013 to may 2014 were divided into IVUS-guided intervention therapy group (n=89) and coronary angiographyguided group (n=90).Their baseline parameters, lesion features, MACE and restenosis during follow-up were analyzed.Results Baseline clinical and angiographic characterisitcs were well matched and show no significant differences between the two groups.Compared to angiography-guided group, the minimum lumen diameter (MLD) of IVUS-guided group were higher, diameter stenosis (DS) and lesion length (LL) were lower in IVUS-guided intervention therapy group.The detection rate of calcified and eccentric lesion were significantly higher in IVUS-guided group.There were 204 (93.6 ％) and 195 (87.8％) high-pressure balloons used in post-inflation in two groups, respectively.The MLD and plaque burden of IVUS-guided group were obviously improved after stent implantation.Conclusions IVUS-guided intervention therapy in PCI is safe and effective, may be helpful for the judgment of lesion, evaluating stent implantation and guiding high-pressure balloon post-inflation.IVUS-guided intervention could get the bigger immediate lumen diameter and lower plaque burden than coronary angiography without serious short-term or long-term complications.

Objective To evaluate the residual risk (i.e.failure risk in detecting aneuploidies abnormalities except for chromosome 13,18,21,X and Y) of cytogenetic abnormalities using interphase fluorescence in situ hybridization (FISH) for the second-trimester amniocytes.Methods The results of interphase FISH and conventional karyotyping of 2 837 consecutive amniotic fluid specimens were analyzed retrospectively.Probes for chromosomes 13,18,21,X and Y were used.The detection rate and residual risk for interphase FISH were calculated for the following three major clinical indications for prenatal diagnosis (advanced maternal age,abnormal maternal serum screening indicating an increased risk for trisomy 18 or trisomy 21,and ultrasound abnormalities).Results Consecutive interphase FISH and karyotyping of second-trimester amniocytes for prenatal diagnosis were performed from January 1,2010 to July 31,2013.Among the 2 837 cases,85 (3.0％) cases with abnormal karyotypes were found,including 73 cases of aneuploidies involving chromosome 13,18,21,X and Y,which were considered detectable by interphase FISH; 12 cases of chromosomal anomalies,other than aneuploidies of chromosome 13,18,21,X and Y,were diagnosed after karyotyping and were not detected by interphase FISH,including six cases of balanced rearrangements,five cases of imbalanced rearrangements,and one case of pseudomosaic of trisomy 20.Of these 12 chromosomal anomalies,three cases of imbalanced rearrangements involving chromosome 21 showed positive FISH results,and the other nine cases showed negative FISH results among which four case of hereditary balanced rearrangemerts and two cases of novel balanced rearrangements.The total detection rate for interphase FISH was 89.4％ (76/85),the misdiagnosis rate of chromosome abnormalities was 14.1％(12/85),and the residual risk was 0.43％ (12/2 761) following interphase FISH of the second-trimester amniocytes.Conclusions Interphase FISH is a useful adjunct to conventional karyotyping,but should not be regarded as a replacement for karyotyping as too many structural chromosomal abnormalities will be missed.Providing patients with a detection rate and residual risk during counselling may help them understand the advantages and limitations of interphase FISH in their prenatal diagnostic evaluation.

Using cyproheptadine ( CYP) as template molecule, methacrylic acid ( MAA) as monomer, ethylene glycol dimethacrylate (EGDMA) as cross-linker, molecularly imprinted polymers (MIP) with high selectivity to cyproheptadine (CYP) were prepared by the optimization of porogen, monomer, and the mole ratio of monomer to template. The specific surface area of the prepared polymers was 24. 9 m2 / g. The recovery of CYP was above 94. 0% when the following procedure was applied to the cartridge of MIP as adsorptive material: conditioning with methanol and water, loading with water, washing with water and methanol, and eluting with methanol-ammonia (95: 5, V/ V). As a control, the recovery of CYP on non-imprinted polymers cartridge (NISPE) was only 38. 9% . The binding capacity of the molecularly imprinted solid phase extraction (MISPE) towards CYP found to be about 8. 8 mg of CYP/ g polymers and the imprinting factor (IF) was about 2. 32. Under optimal conditions, a mixed standard solution of CYP, amitriptyline, sulfadiazine and trimethoprim (10 mg / L each) was uploaded on the MISPE and NISPE for selectivity experiment. The gradient elution was used by using 0. 05% sodium pentanesulfonate solution (A)-acetintrile (B) as a mobile phase. The recoveries on the MISPE for sulfadiazine and trimethoprim (different structure with CYP) were less than 10% , however, the recovery for the similar structural amitriptyline was more than 70% , and the recovery more than 90% for CYP. All the recoveries on the NISPE for four analytes were less than 30% . This new MISPE cartridge was applied to extract and enrich CYP in livestock drinking water sample, and the recoveries of CYP ranged from 80. 5% -97. 7% , and the limit of detection (LOD) was 0. 01 mg / L.