Diclofenac potassium delayed-sustained release pellets were prepared by double-layer coating method with ethylcellulose aqueous dispersion.The effects of release condition and pellet compositions on the in vitro drug release were evaluated.The formulation was optimized by the central composite design-response surface methodology.It was shown that the pH of the media greatly affected the in vitro drug release of the pellets while the viscosity of the media had little influence.Drug release from the pellets was related to the proportion of the inner coat to the outer coat and the amount of pore forming agent in the outer coat.The optimization of the formulation could be achieved by the central composite design-response surface methodology.

OBJECTIVE:To provide reference for medical costs control in China. METHODS:Based on main characteristics of the United States health service system,information asymmetry,principal-agent theory and Freedman’s consumption theory were used to analyze the advantages of restrictive relationship among doctors,hospitals and health insurance institutions in control-ling medical costs and improving the quality of diagnosis and treatment. The growth rate of medical cost,the percentage of drug ex-penditure and other aspects were compared between China and the United States;the effect of restrictive relationship on medical cost control was demonstrated. RESULTS & CONCLUSIONS:In the United States,there are hierarchical medical system and two-way referral system;for-profit hospitals and non-profit hospitals are mutually complementary;different natures of health insur-ance system,different payment methods and strict“commercial bribe”monitoring system are carried out;doctors,hospitals and health insurance restrict each other. Not only there are many advantages in theory,but also in practice the growth rate of medical cost and the percentage of drug expenditure are superior to our country. Finally it controls the rapid growth of medical cost to a cer-tain extent. Combined with our national conditions,learning from the United States experience,restrictive relationship among doc-tors,hospitals and health insurance institutions is established to control the increase of medical cost in China through reducing infor-mation asymmetry and standardizing payment audit;establishing a scientific pattern of mixed payment;strengthening the indirect impact of the health insurance institutions on doctors and hospitals,etc.

Both sides of the picornavirus genome have 5'-untranslated region (5'UTR) and 3'- untranslated region (3'UTR). This study demontrated that both the 5'-and 3'-UTR can form complex structures, such as stem-loop, clover and pseudoknot structure, These structures play an important role in the regulaton of the replication and translation of the viruses. This article reviewed the progress of research on the structure and function of picornavirus' 3'-UTR over recent years.
3' Untranslated Regions ; Animals ; Humans ; Nucleic Acid Conformation ; Picornaviridae ; chemistry ; genetics ; metabolism ; Picornaviridae Infections ; virology ; RNA, Viral ; chemistry ; genetics ; metabolism

Objective To observe the therapeutic effect of microencapsulated human retinal pigment epithelial (RPE) cell transplantation into the striatum in a rat model of Parkinson' disease (PD). Methods Cultured RPE cells were microencapsulated by alginate-polylysine-alginate (APA) using a high voltage electrostatic system. The Parkinsonian rats were divided into 4 groups, namely the model group, RPE group, APA group and RPE-APA group, and in the latter 3 groups, RPE cells, empty APA microcapsules and APA-capsulated RPE cells, respectively, were transplanted into the right striatum of the rats via stereotactic surgery. After the transplantation, the changes in apomorphine-induced rotation of the rats were investigated and the striatum DA contents were measured with high-performance liquid chromatography with electrochemical detection. Results TThe rotation of the rats in RPE-APA group was reduced by 51.48%, showing significant difference from that in the model control group 4 weeks after the transplantation (P<0.05). At 8 weeks after the transplantation, apomorphine-induced rotation in the RPE-APA group showed further significant reduction by 54.43% (P<0.05). The changes of DA contents in the striatum after transplantation were consistent with the changes in the rotation behavior of the rats. Conclusions Microencapsulated human RPE cell transplantation is a promising approach for the treatment of PD.

Objective To evaluate the value of using a linear stapler device for the cloure of the pharynx during total laryngectomy.Methods Sixteen total laryngectomies were performed between August 2010 and December 2011,during the operation,the TA 60 linear stapler was used for pharyngeal closure.Among these patients,two patients had the history of pre-operative radiotherapy,four patients recurred after radiotherapy,ten patients were treated for the first time.100 ml methylene blue was injected into the newly closed laryngopharyngeal cavity through the nasopharyngeal breather pipe for checking up whether it was watertight or not.Results Amnong the sixteen patients,methylene blue leakage from the mucosal joint of the gular cavity closed by the stapler were not found in fifteen patients,it was only found in one patient.The transudatory places were sutured with absorbable Vicryl sutures. This patient healed well without pharyngocutaneous fistula.Negative surgical margins were achieved in all patients.No patient needed to be tranfered to open surgery.Using a linear stapler device in total laryngectomy,45 minutes could be saved as compaired to manual suture. One patient developed a light pharyngocutaneous fistula.The incidence of pharyngocutaneous fistula was 6.25％ (1/16). Conclusions This stapled closed technique for pharyngoplasty is efficient,eliminates the risk of wound contamination,saves operation time and decreases the incidence of pharyngocutaneous fistula.This technique can be recommended as alternative for repairing the pharynx in patients undergoing total laryngectomy.

Objectives To construct the cancellous bone explant model and a method of culturing these bone tissues in vitro, and to investigate the effect of mechanical load on growth of cancellous bone tissue in vitro.
Methods Cancellous bone were extracted from rabbit femoral head and cut into 1-mm-thick and 8-mm-diameter slices under sterile conditions. HE staining and scanning electron microscopy were employed to identify the histomorphology of the model after being cultured with a new dynamic load and circulating perfusion bioreactor system for 0, 3, 5, and 7 days, respectively. We built a three-dimensional model using microCT and analyzed the loading effects using finite element analysis. The model was subjected to mechanical load of 1000, 2000, 3000, and 4000μεrespectively for 30 minutes per day. After 5 days of continuous stimuli, the activities of alkaline phosphatase (AKP) and tartrate-resistant acid phosphatase (TRAP) were detected. Apoptosis was analyzed by DNA ladder detection and caspase-3/8/9 activity detection.
Results After being cultured for 3, 5, and 7 days, the bone explant model grew well. HE staining showed the apparent nucleus in cells at the each indicated time, and electron microscope revealed the living cells in the bone tissue. The activities of AKP and TRAP in the bone explant model under mechanical load of 3000 and 4000μεwere significantly lower than those in the unstressed bone tissues (all P<0.05). DNA ladders were seen in the bone tissue under 3000 and 4000μεmechanical load. Moreover, there was significant enhancement in the activities of caspase-3/8/9 in the mechanical stress group of 3000 and 4000με(all P<0.05).
Conclusions The cancellous bone explant model extracted from the rabbit femoral head could be alive at least for 7 days in the dynamic load and circulating perfusion bioreactor system, however, pathological mechanical load could affect the bone tissue growth by apoptosis in vitro. The differentiation of osteoblasts and osteoclasts might be inhibited after the model is stimulated by mechanical load of 3000 and 4000με.

BACKGROUNDIt is generally accepted that spleen plays a complex role in the tumor immunity, which would change in the different periods of cancer. In this study, we investigated the changes in the function of splenic macrophage (Mphi) in different stages of liver cancer induced by diethylnitrosamine (DEN) in rats. The aim was to support the characteristics of "two-way" and "phase" of spleen in tumor immunity.

METHODSThe model of pulmonary metastasis of liver cancer was established in forty male SD rats by DEN. In the 8th, 13th and 16th week, 10 rats were randomly chosen and sacrificed, and divided into cirrhosis, liver cancer and pulmonary metastasis groups depending on the pathological result, respectively. The other 10 rats were taken as control group. The Mphi was isolated by anchoring cultivation. The changes in ultrastructure, phagocytosis, cytokine secretion, antigen processing and presenting, and viability of splenic Mphi were detected by transmission electron microscopy, Vybrant(TM) Phagocytosis Assay, DQ(TM) Ovalbumin, and rat TNF-alpha ELISpot kits.

RESULTSUnder the electron microscope, the Mphi in the control group had some pseudopodium-like prominences, and mitochondria, ribosome, rough endoplasmic reticulum, lysosome can be found in the cytoplasm, and phagocytized RBC. In the liver cirrhosis and liver cancer group, Mphi had more prominences, meanwhile much more mitochondria, ribosome, rough endoplasmic reticulum, lysosome can be found in the cytoplasm, especially in the liver cancer group. In the pulmonary metastasis group, the Mphi was swelling, with few organelle. As compared to the control group, the function of splenic Mphi increased in cirrhosis and cancer groups, but decreased in metastasis group (phagocytosis rate: (84.7 +/- 1.9)%, (89.5 +/- 3.1)%, and (36.0 +/- 2.6)% vs (75.6 +/- 1.7)%, P < 0.05, P < 0.01; viability: (1.53 +/- 0.15)%, (1. +/- 0.14)%, and (1.12 +/- 0.29)% vs (1.48 +/- 0.17)%, P < 0.05, P < 0.01; TNF-alpha secretion: (741.0 +/- 52.9)%, (1126.2 +/- 174.5)%, and (313.8 +/- 50.8)% vs (626.6 +/- 24.6)%, P < 0.05, P < 0.01; positive cell rate of antigen processing and presenting: (24.03 +/- 1.87)%, (27.95 +/- 2.63)%, and (10.46 +/- 2.16)% vs (16.45 +/- 1.86)%, P < 0.01).

CONCLUSIONSIn the stage of cirrhosis and early cancer, the immune functions of splenic Mphi were reinforced. It may promote the non-specificity tumor immunity. On opposite, in the stage of pulmonary metastasis, the immune functions of splenic Mphi were impaired. It may lead to the decrease of tumor immunity.

Animals ; Cells, Cultured ; Diethylnitrosamine ; toxicity ; Disease Models, Animal ; Liver Cirrhosis ; immunology ; pathology ; Liver Neoplasms, Experimental ; chemically induced ; complications ; immunology ; ultrastructure ; Lung Neoplasms ; immunology ; secondary ; ultrastructure ; Macrophages ; pathology ; ultrastructure ; Male ; Microscopy, Electron, Transmission ; Rats ; Rats, Sprague-Dawley ; Spleen ; pathology ; ultrastructure

Animals ; Binding Sites ; Blood Glucose ; metabolism ; Diabetes Mellitus, Type 2 ; blood ; Enzyme Activation ; drug effects ; Enzyme Activators ; chemistry ; pharmacology ; Glucokinase ; chemistry ; metabolism ; Humans ; Hypoglycemic Agents ; chemistry ; pharmacology ; Molecular Conformation ; Phosphorylation ; drug effects ; Sulfones ; pharmacology ; Thiazoles ; pharmacology

It review the structure-function relationship of natural anthraquinone derivatives from Radix et Rhizoma Rhei. The anthraquinone derivatives had many identical activities because they have the identical mother nucleus; but the strength of their activities were different, because they have different substitution groups. The anthraquinone derivatives shown the obvious structure-function relationship in many respects, such as antioxygenation, antibiosis, anticancer, the influence of immunity and so on.
Anthraquinones ; chemistry ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Molecular Structure ; Rheum ; chemistry ; Structure-Activity Relationship

OBJECTIVETo establish fluorescence resonance energy transfer (FRET) assay method of detecting proteolytic activity of non-structural protein 3-4A (NS3-4A) serine protease of hepatitis C virus (HCV) for high throughput screening inhibitors against HCV in vitro.

METHODSHCV recombinant plasmid pMAL~c2/NS3-4A was transformed into the E.coli strain K12TB1. Maltose-binding-protein (MBP) NS3-4A fusion protein expression was induced by adding isopropyl-β-D-thiogalacto-pyranoside (IPTG) and purified by affinity chromatography. The proteolytic activity of MBP-NS3-4A protease was analyzed by FRET with the special protease substrate. The reaction system in this model was optimized, and the reliability of the model was evaluated.

RESULTSHigh throughput screening model for HCV NS3-4A protease inhibitors was established, and the best concentrations of enzyme and substrate were optimized. In the model, the Km value of protease was 4.74 μmol/L, Z factor was up to 0.80, and coefficient of variation (CV) was 1.91%. BILN 2061, one of the known HCV protease inhibitors, was measured with the Ki of 0.30 nmol/L.

CONCLUSIONThe assay model using FRET method for HCV NS3 4A serine protease is stable and reliable, and the model is suitable for high throughput screening for HCV NS3 4A protease inhibitors.

Antiviral Agents ; pharmacology ; Drug Evaluation, Preclinical ; Fluorescence Resonance Energy Transfer ; Hepacivirus ; enzymology ; High-Throughput Screening Assays ; methods ; Protease Inhibitors ; pharmacology ; Viral Nonstructural Proteins ; antagonists & inhibitors ; genetics