Objective: To establish a GC method for the determination of content and content uniformity of memantine hydrochloride dispersible tablets.Methods: The sample was dissolved in water, alkalified by sodium hydroxide solution and extracted by methylene chloride.An HP-5 gas chromatography column (50 m×0.32 mm, 1.05 μm) was used.The column temperature was programming increased, and the initial temperature maintained at 120 ℃ for 3 min, and then raised to 220 ℃ at a rate of 10℃·min-1 and maintained for 7 min.A hydrogen flame ionization detector (FID) was used and the split ratio was 1∶1.The inlet temperature was 230 ℃ and the detector temperature was 260 ℃.The injection volume was 1 μl and the carrier gas was nitrogen with high purity at a flow rate of 3.0 ml·min-1.Adamantane was used as the internal standard, and the internal standard method was used for the calculation.Results: The calibration curve was linear over the range of 0.05-1.0 mg·ml -1 (r=0.999 7).The detection limit and the limit of quantification was 1.1 ng and 3.3 ng, respectively.The average recovery was 100.2% (RSD =0.73%, n=9).Conclusion: The method has the advantages of simple operation, small extraction process toxicity, little environmental pollution, high accuracy and high specificity, and can be used for the determination of content and content uniformity of memantine hydrochloride dispersible tablets.

OBJECTIVE:To establish a method for simultaneous residual determination of dichloromethane and ethyl acetate in bisacodyl raw material. METHODS:Head-space GC was performed on the capillary column of 6% cyanopropyl phenyl-94% di-methyl polysiloxane(DB-624)by temperature programming,the temperature of injector was 220 ℃,detector was flame ionization detector with temperature of 250 ℃,carrier gas was high purity nitrogen with the flow rate of 3.0 ml/min,split ration was 1∶10, headspace heating temperature was 70 ℃,equilibration time was 30 min,volume of headspace vial was 5 ml,and the injection volume was 1 ml. RESULTS:The linear range was 6-120μg/ml for dichloromethane(r=0.999 9)and 50-1 000μg/ml for ethyl ac-etate(r=0.999 9);the limit of quantitation was 0.2,1.7 μg,limit of detection was 0.06,0.5 μg;RSDs of precision,stability and reproducibility tests were no higher than 3%;recoveries were 100.30%-102.00%(RSD=0.63%,n=9) and 100.10 %-101.30%(RSD=0.44%,n=9). CONCLUSIONS:The method is simple and accurate,and can be used for the simultaneous residual deter-mination of dichloromethane and ethyl acetate in bisacodyl raw material.

ObjectiveTo observe the influence of calcium - sensing receptor (CaSR) on Fas/Fas ligand (Fas L) pathway during anoxia/reoxygenation (A/R)- induced cardiomyocytes apoptosis in neonatal rat. MethodsSingle cells were dissociated from minced hearts of 2 - day - old Wistar rats with a 0. 25％ solution of crude trypsin and then cultured as monolayers at a density of 5 x 104cells/cm2 in DMEM medium equilibrated with humidified air containing 5％ CO2 at 37C. Three days after the cells were seeded, the cultured cardiomyocytes were randomly divided into three groups. ( 1 ) control group: cardiomyocytes were continuously cultured for 26 hours in DMEM medium.(2) A/R group: cardiomyocytes underwent anoxia for 2 hours and reoxygenation for 24 hours.(3) GdCl3 group: 300μmol/L GdCl3 was added to the culture medium at the beginning of reoxygenation. Apoptosis of cardiomyocytes was assessed by flow cytometer and morphological alterations were observed with transmission electron microscope.The expression of CaSR, Fas and Fas L were analyzed by Western blot.Results The result of flow cytometer showed that cardiomyocytes apoptosis was 12. 18％ ± 1.54％ in A/R group,and was higher than that in the control group (P ＜0. 01 ). At the same time, mitochondrial cristae dissolution and disappearance could be detected. Compared with A/R group, GdCl3, a specific activator of CaSR, further enhanced cardiomyocytes apoptosis to 20. 25％ + 2. 87％ ( P ＜ 0. 01 ), along with an increment in CaSR, Fas and Fas L expressions ( P ＜ 0. 05 ). ConclusionCaSR is closely involved in cardiomyocytes apoptosia during anoxia/reoxygenation. CaSR could induce apoptosis of neonatal rat cardiomyocytes through Fas/Fas L receptor pathway.

Objective To investigate the effect of 20‐HETE on the isolated myocardial ischemia reperfusion injury and to ex‐plore its underlying mechanisms .Methods Experiments were performed in isolated rat hearts subjected to 35 min of ischemia fol‐lowed by 40 min of reperfusion in Langendorff preparations .HET0016 (1 μmol/L) and various concentrations (10 ,30 or 50 nmol/L) of 20‐HETE were infused 10 min before the onset of ischemia and throughout the reperfusion period .Cardiac hemodynamic changes and myocardial contractility were continuously recorded with the Powerlab /8P system .Myocardial infarct size was meas‐ured by TTC staining .The level of ROS and the protein carbonyl content were determined by DHE fuorescence and DNPH method , respectively .Results Perfusion with HET0016 significantly improved myocardial ischemia reperfusion injury reduction in cardiac contractility ,after inhibited the production of 20‐HETE significantly reduced the occurrence of myocardial infarction area (P<0 .05) ,but exogenous join 20‐HETE aggravated I/R‐induced myocardial injury (P<0 .05) .Myocardial ischemia reperfusion injury significantly increased production of ROS and oxidative stress ,both of which were significantly inhibited by HET 0016 and enhanced by 20‐HETE administration(P< 0 .05) .Conclusion 20‐HETE stimulates ROS production and enhance protein carbonylation , which aggravates myocardial ischemia reperfusion injury .

OBJECTIVE:To investigate the similarity of dissolution curves between generic preparations and reference prepara-tions of Bisacodyl enteric-coated tablets in various dissolution mediums,and to provide reference for improving production technolo-gy and internal quality of generic preparations. METHODS:Paddle method was adopted with rotation speed of 75 r/min. The disso-lution test was performed using 1000 mL pH 6.0 phosphate buffer solution,pH 6.8 phosphate buffer solution,water containing 2% sodium lauryl sulfate. HPLC method was used to determine average accumulative dissolution of main components from 3 kinds of generic preparations and reference preparations at different time points to draw out dissolution curves. Similarity factor(f2)meth-od was used to the similarity of dissolution curves. RESULTS:Dissolution curves of reference preparations were basically the same in 3 kinds of dissolution mediums. But the dissolution curve f2 of one generic preparation among 3 manufactures to dissolution curve of reference preparation were ≥50,namely the similarity. CONCLUSIONS:The quality of generic Bisacodyl enteric-coat-ed tablets produced by different manufacturers is obviously different;the generic preparations needs to be further improved in the production technology and internal quality. For domestic generic preparation,it is necessary to strengthen the real-time monitoring of its dissolution curve,to ensure the drug quality.

The results of analyzing the managerial characteristics and complexity of product cooperative development suggest that task planning is an important aspect for process management of product cooperative development and the method for planning tasks should be able to model the dependency between tasks and iterations during the development process. In this paper, a DSM-based method and its corresponding optimization algorithms are developed. At first the coupled task sets and uncoupled task setsare identified, and the tasks are then optimized by the corresponding algorithms. The optimal tasks plan will reduce the development time and cost. Considering the practical requirements in real world, a Multilayer DSM is proposed, and its information communication techniques between DSM and traversing principle are described in details.

Objective:To analyze the association between CRHR1 gene(rs1876828)polymorphism and the effect of inhaled corticosteroids(ICS)in children with bronchial asthma.Methods:A total of 60 children with moderate persistent asthma who were treated in the Department of Pediatrics of the First Affiliated Hospital of Harbin Medical University from January 2018 to June 2019 were included.The CRHR1 gene rs1876828 locus in children with asthma was detected by Sanger sequencing.The children were divided into TT genotype group(TT group) and CC genotype group(CC group)according to the different base sequences of gene loci.There were 22 cases in TT group(36.7%)and 38 cases in CC group(63.3%). Both groups were given aerosol inhalation of ICS and symptomatic treatment.Clinical symptoms and signs were observed and scored before and after treatment for 3d, 10d and 30d, and the days required for complete disappearance of symptoms and signs were recorded.Results:After 3d of treatment, clinical symptoms and signs of TT group and CC group were improved to a certain extent, but there was no statistical significant difference between two groups( P>0.05). At 10d and 30 d after treatment, the recovery of the two groups was better than that at 3d, and the improvement degree of the TT group was significantly better than that of the CC group, with statistical significance( P<0.05). The time of complete remission of symptoms and signs in TT group and CC group was(8.68±7.42)d and(16.21±7.82)d; the difference was statistically significant( P<0.01). Conclusion:There is a polymorphism of CRHR1 rs1876828 locus in children with bronchial asthma, which manifests as TT genotype and CC genotype, and CC genotype is the majority.The polymorphism of CRHR1 gene rs1876828 in asthmatic children is associated with the efficacy of ICS.The efficacy of ICS in children with TT genotype is better than that of CC genotype.

Objective:To evaluate the measurement uncertainty in the determination of sodium valproate tablets by GC with an in -ternal standard method , and determine the main sources of uncertainty .Methods:A GC internal standard method was selected to sys-tematically analyze the uncertainty in the determination of sodium valproate tablets , including the sample quantity , dilution ratio, purity and area repeatability of chromatographic peaks .Results: The expanded uncertainty of sodium valproate tablets was 2.7%, and the determination range of sodium valproate tablets was (96.3 ±2.7)%(k=2).Conclusion:The established GC internal standard meth-od for the uncertainty evaluation is reliable , which is helpful to improving the quality evaluation and control of sodium valproate tablets .

AIM To observe the effects of Huangdi Anxiao Capsules (Puerariae lobatae Radix,Eriobotryae Folium,Notoginseng Radix et Rhizoma,etc.) on blood glucose and insulin resistance in GK rats with type 2 diabetes mellitus (T2DM) and its possible mechanism of action.METHODS GK rats with T2DM included in the experiment were randomly divided into model group,rosiglitazone (1.44 mg/kg) group,Shenqi Jiangtang Granules (1.08 g/kg) group,Huangdi Anxiao Capsules high,medium and low (12,6,3 g/kg) group and normal control group of Wistar rats.After six weeks of consecutive administration,fasting blood glucose (FBG),serum insulin (FINS),fasting plasma glucose (FPG),and insulin resistance index (HOMA-IR) were measured in all groups.Serum biochemical indexes of (TG,TC,HDL-C,LDL-C),glycosylated hemoglobin (HbA1c),glucagon-like peptide (GLP-1) and insulin-like growth factor (IGF-1) were measured.The pancreatic tissue was obtained by routine paraffin embedding and HE staining to observe the pathological changes.RESULTS Compared with the normal group,FBG,FPG,FINS,HOMA-IR,TG,TC,HDL-C,LDL-C and HbA1c of the model group were significantly higher (P ＜0.01),and GLP-1 and IGF-1 expressions were markedly decreased (P ＜ 0.01).Compared with the model group,the levels of FBG,FPG,FINS,HOMA-IR,TG,TC,HDL-C,LDL-C and HbA1 c in Huangdi Anxiao Capsules high-,medium-and low-dose group were significantly decreased (P ＜ 0.05,P ＜0.01);GLP-1 and IGF-1 expressions were markedly increased (P ＜0.05,P ＜0.01),compared with the model group,the structure changes of pancreatic tissue in Huangdi Anxiao Capsules high-,medium-and low-dose groups largely reduced.CONCLUSION Huangdi Anxiao Capsules can reduce GK rats fasting blood glucose,insulin resistance,and its mechanism may be related to promoting the emergence and proliferation of pancreatic islet cells,improving the function of islet beta cells,and increasing insulin secretion.

BACKGROUND:Smooth muscle cells and transitional epithelial cells were traditionally used to construct tissue-engineered bladder and to perform double-sided implantation of scaffold.However,double-sided implantation is difficult to perform,because smooth muscle cells are difficult to isolate or culture in vitro and passage is limited.OBJECTIVE:To verify the feasibility of tissue-engineered bladder reconstruction with bone marrow mesenchymal stem cells(BMSCs)and bladder acellular matrix(BAM).DESIGN:A basic empirical study.SETTING:Linbaixin Medical Research Center,Second Affiliated Hospital,Sun Yat-sen University.MATERIALS:Experiments were performed at the Linbaixin Medical Research Center,Second Affiliated Hospital,Sun Yat-sen University from March 2006 to Mav 2007.The laboratory was the Opening Laboratory of Hospital Affiliated to Health Department of China.One-month old SD rats of either sex,weighting 80-100 g were provided by Animal Experimental Center of Sun Yat-sen University.Fresh porcine bladders were offered by Animal Experimental Center of Southern Medical University.METHODS:Whole bone marrow culture and successive adherence method was used to culture rat BMSCs in vitro.Flow cytometry was employed to detect surface antigen.Eradicator washing method was applied to prepare porcine BAM and measure its purity and characteristies.Third passage of BMSCs were inoculated in BAM and cultured in a medium containing vascular endothelial growth factor(VEGF165)(25 ng/L)in vive and in vitro to test compatibility.Cells cultured alone were considered to be controls for the in vivo trial,and materials non-implanted with cells were considered to be controls for in vitro trial.Suitable microenvironment was simulated to induce the differentiation of BMSCs.Four weeks and eight weeks later,compound materials were respectively removed to perform tissue section test.Simultaneously,immunohistochemistry keratin staining was conducted to examine regeneration of epithelial cells.MAIN OUTCOME MEASURE:Biocompatibility of BMSCs and BAM.RESULTS:①BMSCs were cultured by whole bone marrow method.Flow cytometry demonstrated that third passage of cells were positive for CD29(99.43%).②BAM had good biological characteristics.Homogen matrix and byssoid collagen appeared under a microscope.Compatibility trials showed good compatibility of BMSCs and BAM and well-growth cells.③Four weeks later,histological section test confirmed inflammatory cell infiltration,closely-arranged collagen and elastic fiber.Immunohistochemistry keratin staining showed lamellar and discontinuous simple epithelium.Eight weeks later,no inflammatory cell infiltration was found,and closely-arranged collagen and elastic fiber were detected.Immunohistochemistry keratin staining showed lamellar and continuous multiple epitheliums.CONCLUSIoN:With good compatibility,BMSCs and BAM appear to be an ideal material for bladder tissue engineering.