OBJECTIVE: To establish a RP-HPLC method for the simultaneous determination of Protocatechuic acid and Protocatechuic aldehyde in Fuxuekang granules. METHODS: The chromatography was carried on Achirom Bond-1 C18 column(250 mm?4.6 mm,5 ?m). The mobile phase was composed of methanol - water (20∶80,pH=2.80 adjusted by glacial acetic acid) at a flow rate of 1.0 mL?min-1. The detection wavelengths were programmed at 256 nm for Protocatechuic acid and at 280 nm for Protocatechuic aldehyde. RESULTS: The linear ranges for Protocatechuic acid and Protocatechuic aldehyde were 5.49～73.22 ?g?mL-1(r=0.999 9) and 4.81～64.13 ?g?mL-1(r=0.999 9), respectively, and their average recoveries were 96.86%(RSD=2.52%,n=9) and 97.55%(RSD=3.82%,n=9) respectively. CONCLUSION: The method is simple, sensitive, and reproducible, and it is applicable for the quality control of Fuxuekang granules.

OBJECTIVE:To establish a method for content determination of cyproheptadine hydrochloride in fuyan cream.METHODS:Ultraviolet spectophotometry was applied to determine the contents of cyprohepatadine hydrochloride as well as its adjuvant as two coexisting components without isolation and extraction,the wavelengths were286nm and258nm res_ pectively,linear regression and simultaneous equations were used to analyze the results.RESULTS:Cyproheptadine hydrochlor_ ide and its adjuvant showed good linear relationship in the range of2.52～25.2and2.5022～25.022?g/ml respectively,the average recovery of cyproheptadine hydrochloride was98.61%(RSD=1.08%).CONCLUSION:The present method is simple,convenient,reproducible and reliable,which is suitable for rapid determination for fuyan cream.

Objective To investigate the expression of Golgi protein?73(GP73) and Ki?67 antigen in gallbladder carcinoma ,and to analyze their correlations with proliferation ,invasion ,and prognosis of gallbladder carcinoma. Methods Streptavid?in?peroxidase (SP) immunohistochemistry was used to detect the expression of GP73 and Ki?67 in surgically resected specimens of 58 gallbladder carcinomas ,15 gallbladder adenomas and 15 gallbladder polyps samples . Results The positive rates of GP73 and Ki?67 protein in gallbladder carcinomas were 72.4% and67.24%,respectively ,which wer significantly higher than those in gallbladder adenomas(GP73:40.0%,Ki?67:26.7%,P<0.05)and in gallbladder polyps(GP73:13.3%,Ki?67:25.0%,P<0.05).The expression of GP73 was positively correlated with tumor differentiation ,Nevin staging and lymph node metastasis(P < 0.05), and the expression of GP73 was positively correlated with tumor differentiation ,Nevin staging(P < 0.05). GP73 expression was positively correlated with Ki?67 expression in gallbladder carcinoma (r = 0.473 ,P = 0.000). Patients with negative expression of GP73 and Ki?67 had longer survival time than those with positive expression of GP73 and Ki?67. Conclusion The expression of GP73 and Ki?67 was associated with proliferation ,invasion of gallbladder carcinoma. The combined detection of GP73 and Ki?67 is conducive to judging the progress and prognosis of the gallbladder carcinoma .

Measurements of the magnetic properties and total contents of Cu, Cd, Pb and Fe in 30 automobile emission particulate samples indicated the presence of magnetic particles in them. The values of frequency dependent susceptibility (chi(fd)) showed the absence of superparamagnetic (SP) grains in the samples. The IRM(20 mT) (isothermal remanent magnetization at 20 mT) being linearly proportional to SIRM (saturation isothermal remanent magnetization) (R(2)=0.901), suggested that ferrimagnetic minerals were responsible for the magnetic properties of automobile emission particulates. The average contents of Cu, Cd, Pb and Fe in automobile emission particulates were 95.83, 22.14, 30.58 and 34727.31 mg/kg, respectively. Significant positive correlations exist between the magnetic parameters and the contents of Pb, Cu and Fe. The magnetic parameters of automobile emission particulates reflecting concentration of magnetic particles increased linearly with increase of Pb and Cu content, showed that the magnetic measurement could be used as a preliminary index for detection of Pb and Cu pollution.
Algorithms ; Environmental Monitoring ; methods ; Feasibility Studies ; Magnetics ; Metals, Heavy ; analysis ; chemistry ; Statistics as Topic ; Vehicle Emissions ; analysis

Objective To study the expression and the clinical significance of hypoxia-induced factor-1α (HIF-1α) in gallbladder cancer tissues,and the role and mechanism of HIF-1α in metformin-suppressed metastasis in gallbladder carcinoma GBC-SD cells.Methods 24 specimens of gallbladder cancer tissues and 5 specimens of chronic cholecystitis were collected from the First Affiliated Hospital of Zhengzhou University between June 2016 and February 2017.Immunohistochemistry and qPCR were used to detect the expression of HIF-1α in gallbladder cancer tissues,in adjacent non-cancer tissues and in chronic cholecystitis,and the clinical significance was analyzed.The model of metastasis was induced by hypoxia;the wound healing assay and the Transwell assay were used to detect the ability of cell metastasis;the expressions of HIF-1α and VEGF in gallbladder carcinoma GBC-SD cells were detected by western blotting assay and immunofiuorescence.Results The expression of HIF-1α in gallbladder cancer tissues was higher than the adjacent non-cancer tissues and in chronic cholecystitis.The expression of HIF-1α was correlated with lymph node metastasis and TNM staging in gallbladder cancer tissues (P ＜ 0.05).The wound healing rate after 48 h in the negative control group and in the treatment with hypoxia group (1％ O2) in GBC-SD cells were (46.5 ± 4.8) ％ and (67.3 ± 4.0) ％,respectively.The Transwell data showed that the numbers of metastasis after 24 h in the negative control group and in the treatment with hypoxia group GBC-SD cells were (147.4 ± 11.7) and (234.4 ± 17.7),respectively.When compared with the negative control group,treatment with hypoxia significantly increased the ability of metastasis and up-regulated the expression of HIF-1α and VEGF in GBC-SD cells (P ＜ 0.05).The wound healing rate after 48 h in the negative control group,the metformin group,the hypoxia group and the metformin and hypoxia group in GBC-SD cells were (40.6 ± 7.1) ％,(16.4 ± 9.4) ％,(69.5 ± 4.0) ％ and (22.4 ± 7.4) ％,respectively.The Transwell data showed that the numbers of metastasis after 24 h in the negative control group,the metformin group,the hypoxia group and the metformin and hypoxia group in GBC-SD cells were (148.4 ± 6.9),(90.0 ± 8.4),(185.8 ± 10.2) and (113.4± 8.6),respectively.When comparcd with the hypoxia group,treatment with metformin and hypoxia significantly decreased the ability of metastasis and down-regulated the expression of HIF-1α and VEGF in GBC-SD cells (P ＜ 0.05).The wound healing rate after 48 h in the negative control group,the 2MeoE2 group,the hypoxia group,the 2MeoE2 and hypoxia group in GBC-SD cells were (43.4 ±4.4)％,(25.9 ±9.0)％,(63.3 ±2.2)％,(46.2 ±4.5)％,respectively.The Transwell data showed that the numbers of metastasis after 24 h in the negative control group,the 2MeoE2 group,the hypoxia group,the 2MeoE2 and hypoxia group in GBC-SD cells were (144.2 ± 12.6),(80.2 ±7.7),(203.8 ±7.0),(124.0 ± 5.2),respectively.When compared with the hypoxia group,treatment with HIF-1α inhibitor 2MeoE2 and hypoxia significantly decreased the ability of metastasis and down-regulated the expression of HIF-1α and VEGF in GBC-SD cells (P ＜ 0.05).Conclusions The expression of HIF-1 α was correlated with lymph node metastasis and TNM staging in gallbladder cancer tissues.Treatment with hypoxia significantly increased the expression of HIF-1α and VEGF and promoted metastasis of GBC-SD cells,while treatment with metformin decreased the ability of metastasis induced by hypoxia via inhibiting the HIF-1o/VEGF pathway in GBC-SD cells.

Aim To observe the effects of tetrahydro-biopterin ( BH4 ) on nitric oxide ( NO ) production in the kidney of type 2 diabetic nephropathy ( DN) mice, and to find a new target for the treatment of type 2 DN. Methods The 12 week-old db/db mice developed in-to DN phase were divided into 2 groups:DAHP group, subjected to intraperitoneal injection of 150 mg·kg-1 DAHP (n=8);DN group, subjected to intraperitone-al injection of same dose of normal saline containing 5% DMSO ( n = 6 ) . The age-matched db/m mice ( NS group) were subjected to intraperitoneal injection of same dose of normal saline containing 5% DMSO ( n =6 ) . Three groups of mice were treated for 7 days. Then the fasting blood-glucose, serum creatinine, u-rine protein and activity of iNOS were determined by chemical colorimetry. And the iNOS protein in renal cortex was determined by immunohistochemisty and western blot, respectively. BH4 was measured by HPLC method. NO level was determined by Griess method. Results The levels of fasting blood-glucose, serum creatinine, 24h urine volume, 24h urine pro-tein, BH4 , iNOS and NO in DN group were signifi-cantly higher than those in NS group;The levels of ser-um creatinine, urine volume, urine protein, BH4 , iN-OS and NO in DAHP group were significantly lower than those in DN group. Conclusion In the kidney of type 2 DN mice, the increased BH4 contributes to over-production of NO by the increased iNOS expression, and resultes in the increase of urine volume and urine protein.

Objective To investigate the relationship between cathepsin L and apoptosis cell in rats after cerebral ischemia reperfusion.Methods Sixty healthy male Sprague-Dawley Rats (10-12 weeks old,260-300 g) were chosen.Based on the random number table method,the rats were randomly divided into sham-operated control group (Sham group,n =10),ischemia-reperfusion group (model group,n =25),and Z-FY-DMK intervention group (CLI group,n =25).Rats were randomly divided into 6 h,12 h,24 h,and 48 h four subgroups in model group and CLI group,respectively.Modified transient middle cerebral artery occlusion was made as Longa described,the intervention groups were injected intracerebroventricularly Z-FY-DMK (20 μg / 1μ1 ×5 μl) preoperative 30 min prior to surgery,Sham group and schemia reperfusion injury (IRI) group were injected intracerebroventricularly dimethyl sulfoxide (DMSO) 5 μ1 (10ml/L) at the same time.Cell apoptosis was detected by terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) straining.Western blotting was used to detect the expression of cathepsin L and caspase-3.Results In the cortical area of ischemic brain,apoptosis cells of sham operation group were rare,while apoptosis of nerve cells of model group with 6 hours reperfusion were visible,and were gradually increased in the order of 12 hours,24 hours and 48 hours.At the same time point,the apoptosis cells of CL intervention group (6 h,12 h,24 h,48 h) were obviously less than model group (P ＜0.05).Western blotting found little visible cathepsin L protein expression in ischemic cerebral cortex preoptic in the sham group.For model group,the cathepsin L expression initially increased in sub groups with 6 hours reperfusion,reached to a peak in sub groups with 12 hours and 24 hours,and remained a high level in sub groups with 48 hours reperfusion.Compared to model group,the cathepsin L expressions of CL intervention group were obviously decreased at all time points (P ＜ O.05).Conclusions Cathepsin L may be involved in neuronal apoptosis by means of caspases 3 pathway.

We recruited consecutive patients with stage III epithelial ovarian, tubal, and peritoneal cancers who had optimal residual tumor after primary cytoreductive surgery and who received intraperitoneal chemotherapy between 2002 and 2012. Two propensity score?matched sample cohorts were created. We found that the addition of paclitaxel as a second intraperitoneal agent on a 3?week dosing schedule did not yield significant incremental survival benefits over the intraperitoneal delivery of a single cisplatin?based regimen. If our findings could be confirmed by a prospective randomized study, then it would be interesting to explore the efcacy of shifting back to a dose?dense intraperitoneal delivery of paclitaxel or a dose?dense delivery of a new formulation of paclitaxel for the patients with stage III epithe?lial ovarian, tubal, and peritoneal cancers.

Physiologically based pharmacokinetic (PBPK) modeling and simulation can be used to predict the pharmacokinetic behavior of drugs in humans using preclinical data. It can also explore the effects of various physiologic parameters such as age, ethnicity, or disease status on human pharmacokinetics, as well as guide dose and dose regiment selection and aid drug-drug interaction risk assessment. PBPK modeling has developed rapidly in the last decade within both the field of academia and the pharmaceutical industry, and has become an integral tool in drug discovery and development. In this mini-review, the concept and methodology of PBPK modeling are briefly introduced. Several case studies were discussed on how PBPK modeling and simulation can be utilized through various stages of drug discovery and development. These case studies are from our own work and the literature for better understanding of the absorption, distribution, metabolism and excretion (ADME) of a drug candidate, and the applications to increase efficiency, reduce the need for animal studies, and perhaps to replace clinical trials. The regulatory acceptance and industrial practices around PBPK modeling and simulation is also discussed.

Objective To develop the UHPLC-MS/MS method for the determination of amygdalin, paeoniflorin, ferulic acid, calycosin glucosidase, quercetin and formononetin in Buyang-Huanwu decoction. Methods Isocratic elution was carried out with mobile phase consisting of methanol- 2 mM ammonium formate. The separation was performed on Agilent ZORBAX SB-C18 maintained at 35 ℃. The flow rate was 200 μl/min, and the injection volume was 2 μl. The mass spectrometer was operated in the positive and negative ionization electrospray (ESI) mode using multiple monitoring (MRM) for analysis of six components. The mass spectrometric conditions were that ion source temperature 400 ℃, dry gas flow 500 L/h, atomization gas flow rate 75.8 Kpa, spray voltage 4000 V, dry gas temperature 400 ℃. Results The amygdalin, paeoniflorin, ferulic acid, calycosin glucosidase, quercetin and formononetin were all analyzed exactly, and the linear ranges were 0.5-32, 0.2-12.8, 0.1-6.4, 0.8-51.2, 0.4-25.6, 0.08-5.12 ng, respectively. The r were 0.9921, 0.9945, 0.9928, 0.9958, 0.9947, 0.9966, respectively. The recoveries of six analytes ranged from 99.21％ to 101.44％ and the relative standard deviations were all below 2.05％. Conclusions A sensitive, accuracy and suitable UHPLC-MS/MS method has been developed, and the method could be applied for the determination of amygdalin, paeoniflorin, ferulic acid, calycosin glucosidase, quercetin and formononetin in Buyang-Huanwu decoction.