Objective:To collect the bioburden information in clean room to understand the bioburden status, find out weak points and risks in microbial control and improve the management efficiency in clean room. Methods: According to GB/T 16293-2010 and the standard operation practice ( SOP) in our lab, the bioburden information was obtained by the collection and identification of air-borne microbe and surface bacteria in the four main areas of clean room ( microbial limit test room, sterile room 1 and 2, positive room) and on the person entered clean room. Results:The preliminarily established bioburden information indicated that the main mi-croorganism in clean room was Micrococcus and Staphylococcus. The detection rate of fungi was about 5% in clean room. Conclusion:The movement of people and goods in clean room should be strengthened, and samples should be with thorough disinfection.

Objective To construct and identify recombinant expression plasmid of small interfering RNA (siRNA)targeting hepatitis B virus X protein(HBx), and observe its effect on mitoehondrial function in healthy liver cell line steadily expressed HBx gene (HL-7702/HBx). Methods Two siRNA sequences containing short hairpin structure, which target on the total length HBx gene, were synthesized and cloned into the vector psiRNA-Hh1GFPzeo to eonstruct recombinant expression plasmids pX1 and pX2. Non-specific recombinant pScr plasmid served as control. After siRNA transfected into HL-7702/HBx cells line by liposome, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were performed to identify the suppressive effect on HBx expression. Levels of intraeellular reactive oxygen species (ROS) and mitochondrial membrane potential (△ m) were determined by flow cytometry. The experimental results were compared by analysis of variance. Results Successful constructions of pX1 and pX2 were confirmed by restriction enzyme digestion and sequencing. The expressions of HBx mRNA and protein after 48 h of transfection into HL-7702/HBx cells in control group were 0.65± 0.12 and 0.62± 0.09, respectively, which were both higher than those (0.33±0.10 and 0.19±0.08, respectively) in group pX1 (t=4.73, P<0.05; t=7.53, P<0.05) and those (0.48±0.10 and 0.37±0.11, respectively) in group pX2 (t=2.39, P<0.05;t=4.43,P<0.05). But the inhibition of group pX1 was stronger than that of pX2 (t=2.28,P<0.05). Levels of ROS and △ m after RNA interference were 5.00±0.38 and 33.86±0.50, respectively, while those in control group were 72. 10±0. 55 and 3. 57±0.26, respectively (ROS: t=276.22, P<0.05; △ m: t=107.15, P<0.05). Conclusions siRNA targeting HBx can efficiently and specifically suppress the HBx expression in HL-7702/HBx cells, and decrease the level of ROS and increase the level of △ m, thus relieve cellular oxidative stress.

The aim of the present study was to increase distribution of yuanhuacine in the lungs and achieve the purpose of reducing toxicity and increasing efficiency.Therefore,yuanhuacine was designed to be dry powder inhalers innovatively and directly delivered to the lungs.Accordingly,inhaled lactose was used as a carrier to adsorb yuanhuacine on the surface of lactose.Fine particle fraction (FPF) was utilized as evaluation index to filtrate the optimal prescription for pulmonary administration.Besides,an UHPLC-MS/MS method was established for the analysis of heart,liver,spleen,lung,kidney,brain and reproductive system of rats.Intravenous injection was taken as reference to investigate the distribution of yuanhuacine and calculate relevant targeting parameters.The experimental result indicated that the prescription (rough lactose ∶fine lactose =10 ∶ 1) has the highest FPF,which can be chosen as the most suitable prescription for pulmonary administration of yuanhuacine.Moreover,by comparing the distribution of yuanhuacine through pulmonary administration and intravenous injection,it was found that the concentration of yuanhuacine in the lung tissue was greatly increased by pulmonary administration,which decreased the distribution in heart,liver,spleen,kidney,brain and reproductive system,thus sequentially reducing the toxicity in other tissues and increased the efficiency.

Objective To investigate scanning and reconstruction techniques of multi-slice spiral CT (MSCT) in patients with complex congenital heart diseases (CCHD).Methods One hundred eighty-four patients suffering from CCHD underwent 16-detector MSCT scanning without ECG-gating.Multi-planar reconstruction (MPR),maximum intensity projection (MIP),curved-planar reconstruction (CPR) and volume rendering (VR) were used to reconstruct images.CT findings were compared with those of surgical operation or angiocardiography.Results A total of 616 cardiac deformities were found with MSCT and proved by angiocardiograms or surgical operation.The diagnostic accuracy of extracardiac malformation with MSCT was 100%,of atrial septal defect was 54.65%,and of ventricular septal defect was 78.62%.MSCT failed to display heart valve disease well.Conclusion MSCT can accurately detect extracardiac malformations of CCHD.

Objective　To study efficacy of 2,4-dibromo-5, 5-d imethylhydantoin in killing vegetative forms of bacteria and spores of B. subtil is var.niger, and efficacy of 2,4-dibromo-5,5-dimethylhydantoin in dest roying antigenicity of HBsAg. Methods　Neutralizer test and efficacy of so lution of 2,4-dibromo-5,5-dimethylhydantoin in killing vegetative forms of ba cteria and spores. Neutralizer test and efficacy of solution of 2,4-dibromo-5, 5-dimethylhydantoin in destroying HBsAg antigenicity in suspension. Resul ts　The killing rate of Staphylococcus aureus and E. coli. was 100 % a fter exposure to its solution containing 4 mg*L-1 and 2 mg*L-1 av ailable bromo after 10 min and 20 min. The killing rate of spores of Bacil lus subtilis var. Niger also was 100% after exposure to its solution co ntainin g 2 000 mg*L-1 available bromo after 30 min. Its solution containing 1 0 00 mg*L-1 available bromo with could destroy HBsAg in su spension for 5 min. Conclusions　2,4-dibromo-5,5-dimethylhydantoi n can effe ctively kill vegetative forms of bacteria and spores of B. subtilis var.ni ger, and can completely destroy the antigenicity of HBsAg in the water.

The purpose of this study is to investigate the applicability of a natural swelling matrix derived from boat-fruited sterculia seed (SMS) as the propellant of osmotic pump tablets. The sugar components, static swelling, water uptake and viscosity of SMS were determined and compared with that of polythylene oxide (WSR-N10 and WSR-303). Both ribavirin and glipizide were used as water-soluble and water-insoluble model drugs. Then, the monolayer osmotic pump tablets of ribavirin and the bilayer osmotic pump tablets of glipizide were prepared using SMS as the osmotically active substance and propellant. SMS was mainly composed of rhamnose, arabinose, xylose and galactose and exhibited relatively high swelling ability. The area of the disintegrated matrix tablet was 20.1 times as that at initial after swelling for 600 s. SMS swelled rapidly and was fully swelled (0.5%) in aqueous solution with relative low viscosity (3.66 +/- 0.03) mPa x s at 25 degrees C. The monolayer osmotic pump tablets of ribavirin and the bilayer osmotic pump tablets of glipizide using SMS as propellant exhibited typical drug release features of osmotic pumps. In conclusion, the swelling matrix derived from boat-fruited sterculia seed, with low viscosity and high swelling, is a potential propellant in the application of osmotic pump tablets.

In this paper, chloramphenicol was selected as a model drug to prepare in situ gels. The intrinsic dissolution rate of chloramphenicol from in situ gel was evaluated using the surface dissolution imaging system. The results indicated that intrinsic dissolution rate of chloramphenicol thermosensitive in situ gel decreased significantly when the poloxamer concentration increased. The addition of the thickener reduced the intrinsic dissolution rate of chloramphenicol thermosensitive gel, wherein carbomer had the most impact. Different dilution ratios of simulated tear fluid greatly affected gel temperature, and had little influence on the intrinsic dissolution rate of chloramphenicol from the thermosensitive in situ gel. The pH of simulated tear fluid had little influence on the intrinsic dissolution rate of chloramphenicol thermosensitive in situ gel. For the pH sensitive in situ gel, the dissolution rates of chloramphenicol in weak acidic and neutral simulated tear fluids were slower than that in weak alkaline simulated tear fluid. In conclusion, the intrinsic dissolution of chloramphenicol from in situ gel was dependent on formulation and physiological factors. With advantages of small volume sample required and rapid detection, the UV imaging method can be an efficient tool for the evaluation of drug release characteristics of ophthalmic in situ gel.

Objective To establish a method of HPLC assay for determining stilbene glucoside in Zishen Ningshen Pills(ZNP),and to study the influence factors on the content of stilbene glucoside in the process of preparation.Methods HPLC was used for the determination of stilbene glucoside in ZNP.Through simulation the process of preparation,the stilbene glucoside content in the intermediate products was determined by HPLC,and its retention rate and metastasis rate were also investigated.Results The resolution and the linearity of stilbene glucoside were fine,the average recoveries being 98 % ～ 102 %.The retention rate of stilbene glucoside in the drying powder was 60.3 %,lower than that in the original medicinal powder.Conclusion The quantitative method for determining the ingredients in ZNP is simple,feasible and reproducible,and is beneficial for quality control of ZNP.The drying process under normal pressure is the main influence factors of the decrease of stilbene glucoside content,and the decompression drying can be taken into account to take the place of the atmospheric drying.

Objective To investigate the effects of alcohol on hepatitis C virus( HCV) replication and type I interferon signaling pathway in human hepatocytes .Methods Primary hepatocytes were treated with different concentrations of alcohol , and then infected with HCV .The infected cells were collected to measure the level of HCV RNA .The alcohol-treated hepatocytes were also collected to detect the expression of HCV Core, IFN-α, IFN-β, IRF-7, suppressor of cytokine signaling SOCS-2 and SOCS-3 at mRNA and protein levels by real-time quantitative PCR and ELISA or Western blot , respectively .Results Alcohol treatment enhanced HCV infection and replication in primary hepatocytes at concentrations higher than 10 mmol/L in a dose-dependent manner (P<0.05).Treatment with 40 mmol/L of alcohol significantly reduced the expression of IFN-α, IFN-βand IRF-7 at mRNA and protein levels , and increased the expression of SOCS-2 and SOCS-3 at mRNA and protein levels .Conclusion Alcohol treatment could damage the host in-nate immunity in human hepatocytes and promote HCV replication by reducing the expression of type Ⅰinter-feron ( IFN-αand IFN-β) and IRF-7 and increasing the expression of negative regulators including SOCS-2 and SOCS-3.These results demonstrated that the impairment of innate immunity in liver of alcohol abusers might contribute to the enhancement of HCV infection and result in poor therapeutic effect of IFN -α.

Objective To study the phylogenetic evolution and genetic variations of gag gene among the prevalent human immunodeficiency virus (HIV )‐1 strains in Guangxi Zhuang Autonomous Region . Methods Plasma samples of 158 HIV‐1 infected patients in Guangxi area were collected during October 2011 to March 2012 .The gag gene fragments of HIV‐1 were amplified by reverse transcription/nested‐polymerase chain reaction and then sequenced .MEGA 5 .03 was utilized to construct phylogenetic tree and to calculate the genetic distances and selection pressures (globle ω) of gag gene and its coding regions . The comparisons between two groups were tested by Student′s t test ,and the comparisons of multiple groups were tested by one‐way ANOVA .Results A total of 140 amplification products of gag gene were obtained from 158 samples .Four subtypes of HIV‐1 were found ,including CRF01_AE (80 ,57 .1% ) , CRF08_BC (46 ,32 .9% ) ,CRF07_BC (10 ,7 .1% ) ,and subtype B (B′) (4 ,2 .9% ) .The genetic distances of gag gene of the above subtypes were 0 .036 ± 0 .001 ,0 .031 ± 0 .002 ,0 .043 ± 0 .003 and 0 .102 ± 0 .006 ,respectively ,with statistical significance (F=220 .62 ,P<0 .01) .The p17 and p24 coding regions suffered negative selection pressure (globleω<1) .Neither the globle ω in p17 region nor that in p24 region had significant differences among different subtypes (F=0 .761 ,P=0 .469 and F=0 .037 ,P=0 .964 , respectively ) . Conclusion CRF01_AE is the major subtypes of HIV‐1 in Guangxi Zhuang Autonomous Region .The coding regions of gag gene are relatively conserved during evolution .Changes of HIV‐1 prevalence ,however ,may affect the genetic variation of gag gene ,which should be continuously monitored .