OBJECTIVE:The preparation of microspheres was optimized by central composite design in order to improve the lung targeting of tetrandrine chitosa microspheres. METHODS:Microspheres were prepared using an emulsion-chemical crosslink technique.Effects of three independent variables i.e. weight percent of tetrandrine and christon, volume percent of water phase and organic phase and christon concentration in aqueous phase were investigated on four response variables.Response variables selected in the research were yield,drug loading, envelop efficiency, mean diameter and span of dispersity.Second-order polynomial and linear equations were fitted to the data,and the resulting equations were used to produce three dimensional response surface graphs,by which optimal experimental conditions were selected. RESULTS:Five response variables were found to be dependent on three independent variables. According to optimal experimental conditions.An optimized formulation contained weight percent of tetrandrine and christon was 61.97%, volume percent of water phase and organic phase was 13.51% and christon concentration in aqueous phase was 2.37%. CONCLUSION:The central composite design can be used to optimazation of preparation formulation,diameter of the microspheres optimized by it can meet the demands of lung-targeting.

OBJECTIVE:To study the lung targeted characteristic of tetrandrine polylactic acid microsphere(TPM).MET_HODS:TPM and tetrandrine injection were injected intravenously into mouse,an RP-HPLC was established to measure the content of TPM in biological samples,and the concentrations of tetrandrine in different tissues of mice were determined and compared.RESULTS:The determinable concentration of tetrandrine in plasma was in the linear range of 0.519～17.000?g/ml(r=0.9 996),the recovery was 97.32%,the mean value of RSD was 4.46%.After the use of TPM,the concentrations of tetrandrine in mouse tissues were significantly higher than tetrandrine injection,and the highest concentration was detected in lungs of mouse.CONCLUSIONS:TPM is distinguishingly lung targeted.

OBJECTIVE:To provide scientific evidence for the microsphere's m etabolism in vivo of lung targeted tetran?drine polylactic acid(TET-PLA)microspheres.METHODS:Blood was sampled from carotid artery after microspheres were injected into rabbit through auricular vein,the concentration of TET in plasma was determined by RP-HPLC method;The pharmacokinetic parameters were obtained by using3p87program.RESULTS:Concentration-time curves of TET micro?spheres were fitted to a2-compartment model with t 1/2? of(286.49?237.55)min and AUC of(810.33?287.49)(?g?min)/ml.CONCLUSION:TET-PLA microspheres show sustained release effects in rabbits.

OBJECTIVE:To establish a method for the rapid contents determination of potassium chloride and calcium chloride in Compound sodium chloride injection. METHODS:After diluted in appropriate way,Compound sodium chloride injection was sampled directly and contents of potassium chloride and calcium chloride were determined simultaneously by ICP-OES. The powder was 1 300 W,plasma gas flow rate was 15 L/min,auxiliary cooling gas flow was 0.2 L/min,atomizer flow rate was 0.8 L/min, the peristaltic pump rate was 0.8 L/min,atomizer pressure was 315 kPa,and the observation was axial observation,analysis spec-tral lines of potassium and calcium were 766.490 nm and 315.887 nm. RESULTS:The linear ranges of potassium and calcium were 1.0-12.0 mg/L (r=0.999 7 and 0.999 9);RSDs of precision and reproducibility tests were lower than 1%;recoveries were 98.5%-100.5%(RSD=0.59%,n=9)and 99.3%-102.3%(RSD=0.98%,n=9). CONCLUSIONS:The method is simple,rapid and simple,and can be used for the quality control of Compound sodium chloride injection.

Objective To explore the effects and mechanisms of urokinase-type plasminogen activator (uPA) on high glucose-induced rat mesangial cells proliferation and phenotype transformation. Methods Rat mesangial cells were cultured and incubated in media containing either 5 mmol/L D-glucose or 30 mmol/L D-glucose with or without addition of wortmannin, or uPA (105 U/L) for different time periods. At the end of the incubation period, mesangial cells proliferation was assessed by MTT assay and flow cytometric analysis. Cyclin-dependent kinase 2 (CDK2) and p27kip1 expression and activation of Akt were evaluated by Western blotting and Akt kinase assay respectively. Furthermore, the expression and distribution of α-SMA were detected with laser confocal microscopy. Results MTT assay and flow cytometric analysis demonstrated that high glucose induced mesangial cells proliferation (P＜0.05) and an incresed proportion of cells in G2/M+S stage after 24 h incubation (P＜0.01), which were attenuated by uPA or wortmannin (P＜0.01). High glucose induced the enhance of Akt activity after 3 h (P＜0.05), and the effect was inhibited by wortmannin or uPA (P＜0.01). High glucose did not alter CDK2 expression (P＞0.05),but significantly inhibited p27kip1 expression (P＜0.05), which was attenuated by wortmannin or uPA (P＜0.01). High glucose induced the up-regulation of α-SMA expression and perinucleus location in mesangial cells after 24 h (P＜0.01), which were alleviated by wortmannin or uPA (P＜0.01). Conclusion uPA up-regulates p27kip1 expression and counteracts high glucose-induced mesangial cells proliferation and phenotype transformation via blocking PI3K-Akt signaling pathway.

ObjectiveTo investigate the effect of Simvastatin on LOX-1 and ROS in NRK52E induced by ox-LDL.MethodsNRK-52E cells were divided into three groups: Control group,ox-LDL group (50 μg/ml ox-LDL) and Simvastatin group (10 μmol/L Simvastatin +50 μg/ml ox-LDL).After incubation for 24 h,the expression of LOX-I was analyzed by Western blotting,and production of reactive oxygen species (ROS) was analyzed with confocal laser scanning microscopy.ResultsNRK-52E expressed LOX-1 at low level,50 μg/ml ox-LDL increased the expression of LOX-1 by 6.80 times.Pre - treatment with Simvastatin decreased LOX-1 expression by 65％.There was little ROS generation in NRK52E cells,50μg/ml ox-LDL promoted the expression of LOX-1 by 4.86.times.Pre - treatment with Simvastatin decreased ROS generation by 60％.ConclusionsSimvastatin upregulate LOX-1 expression and ROS generation induced by Ox-LDL in NRK52E cells.

Purpose　The aim is to prepare the extract of immunocompetent cell proliferation from human placenta and to try to find out a suitable method of preserving the extract.Methods　The extract was mainly prepared by heat-treating placenta homogenated fluid. Then the activity to stimulate murine splenic lymphocyte proliferation in vitro was done with MTT , exposing　the　extract to　radioisotope 60　Co.Results　The content of protein was 2～3mg per gram placenta measured by Lowry′s Method.The rate to promote cell proliferation was more than 80 percent.The activity lasted a few months after being exposed to radioisotope.Conclusion　The extract prepared by heat-treating not only had high activity, but also had the unique method and good repeatability.This prepared extract as a kind of stable,reliable and remarkably promoting lymphocyte proliferation reaction had the value of development production on broad scale as well as in clinical practice.

Objective To evaluate the effects of rehabilitation training on muscle strength and exercise toleranee in hemodialysis patients with muscle atrophy.Methods Nine hemodialysis patients with muscle atrophy because of end renal failure were recruited in this study. A structured exercise program(90 minutes a sedssion.3 sessions a week)was administered to all the subjects for 6 month.Immediately before and at the end of the exercise programme,the muscle strength of the lower limbs,the motor conduction velocity of the peroneal nerve and maximal oxygen consumption of the patients were examined. Results It was shown that all the patients had impaired exercise capacity,weakend muscle strength and slowed nerve conduction velocity before rehabilitation training.After the exercise programme,the patients' exercise capacity as reflected by the maximal oxygen consumption and exercise time was significantly increased.The muscle strength and the motor nerve conduction velocity were significantly increased.Conclusions Muscle atrophy in hemodialysis patients results in poor exercise tolerance, but rehabilitation exercise programme improves amyotrophy and therefore has beneficial effects on the patient's overall work performance.

Objective To investigate the effect of surfactant protein A (SP-A) on the production of MIP-2 and binding activity of NF-?B in rat tubular epithelial cells, and evaluate its possible role in renal inflammation. Methods Confluent cultures of NRK-52E cells (a renal tubular epithelial cell line of rat origin) were pretreated with various concentrations of SP-A(0 to 80 ?g/ml) and stimulated by lipopolysaccharide (LPS) (10 ?g/ml) with 2% serum. MIP-2 expression was measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). The effect of SP-A on NF-?B binding activity was assessed by electrophoretic mobility shift assay (EMSA). Results MIP-2 mRNA and protein was expressed and up-regulated in NRK-52E cells stimulated by LPS. The expression of MIP-2 was down-regulated by SP-A. NF-?B binding activity was inhibited by SP-A in a concentration-dependent manner. Conclusion SP-A binding activity and down-regulates the expression of MIP-2 in renal tubular epithelial cells, which may play an important role in the modulation of renal tissue inflammation.