In this study, hydrophilic matrix sustained release tablets of total lactones from Andrographis paniculata were prepared and the in vitro release behavior were also evaluated. The optimal prescription was achieved by studying the main factor of the type and amount of hydroxypropyl methylcellulose (HPMC) using single factor test and evaluating through cumulative release of three lactones. No burst drug release from the obtained matrix tablets was observed. Drug release sustained to 14 h. The release mechanism of three lactones from A. paniculata was accessed by zero-order, first-order, Higuchi and Peppas equation. The release behavior of total lactones from A. paniculata was better agreed with Higuchi model and the drug release from the tablets was controlled by degradation of the matrix. The preparation of hydrophilic matrix sustained release tablets of total lactones from A. paniculata with good performance of drug release was simple.
Andrographis ; chemistry ; Delayed-Action Preparations ; chemistry ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; chemistry ; Hydrophobic and Hydrophilic Interactions ; Kinetics ; Lactones ; chemistry ; Tablets ; chemistry

To study still further the activity of CNTF mutant designed by computer molecular modling,the methods of dissociated cultures of chick dorsal root ganglion、TF-1 prolification and the normal mice'weight loss tests weve used.The results indicated that the mutant protein promoted the survival of dorsal root ganglion、induced TF-1 prolification and made the normal mice lose weight,decrease appetite and reduce fat index.The weight loss effect was dependant with its administration dosage,ED50 was 150.986?g/kg/d.To TF-1,the specific activity reached 2.0?106U/mg against international reference reagent.In a word,CNTF mutant had excel bioactivity.So it provided clues for its development and application.

In the present study, the genome shuffling was used to improve lipase production of Penicillium expansum. A lipase producing mutant strain-Penicillium expansum FS8486 and a wild type of Aspergillus Tamarii FS-132 isolated from soil of a volcano in Xinjiang were used as the parental strains. After two rounds of genome shuffling, several elite daughter strains were screened. The lipase activity in one of the daughter strains was increased 317% over the starting strain FS8486. Comparisons of the morphology, RAPD (Random Amplification of Polymorphic DNA) polymorphism and the fatty acid compositions between the daughter and the parental strains suggested that the filial generation were generated by genome shuffling. In this study, the genome shuffling used successfully first time in eukaryotic microorganism and increases the production of the desired metabolite in short time, the study will be useful to spread the genome shuffling in eukaryotic microbial breeding.
Aspergillus ; genetics ; DNA Shuffling ; methods ; Genetic Enhancement ; methods ; Genome, Fungal ; genetics ; Lipase ; biosynthesis ; genetics ; Penicillium ; enzymology ; genetics ; Random Amplified Polymorphic DNA Technique

OBJECTIVETo evaluate application of the sponge of demineralized bone matrix (SDBM) in tissue engineering of bone.

METHODSSDBM was prepared from long bone of rabbits. Bone marrow cells were flushed from the bone shaft of femurs of a two-month-old New Zealand white rabbit. After the cells were cultured for 9 days, the flasks were added into dexamethasone (10(-8) mol/L), beta-glycerophosphate sodium (10 mmol/L) and L-ascorbic acid (50 micrograms/ml). After 5 weeks, the cultured cells were collected and marked by 5-Bromo-2'-dexyouridine (BrdU). The grand sum of cells seeded on a piece of SDBM was about (4-6) x 10(6). The composites of cells and SDBM (tissue engineered chip, TEC) were implanted into muscles and bone defects of radius in rabbits. A standard procedure was applied to make a 10 mm long defect bilaterally in the radius of nine skeletally mature male New Zealand white rabbits. All of the 18 defects were randomly divided into three groups: group I, six defects were grafted by TEC; group II, six defects were grafted with SDBM alone; group III, six defects were empty.

RESULTSThe results of radiographic and histological evaluation showed that all of the defects were repaired in group I and group II at 6 weeks, none of the defects was repaired in group III. The results of BrdU staining showed that the staining was positive in group I, but negative in group II. Biomechanical test showed that the compressive ultimate strength (CUS) of new bone in TEC implanted group was comparable with normal radius (P = 0.623) and in SDBM implanted group was significant lower than normal radius (P = 0.038).

CONCLUSIONSThe TEC can form cartilage and bone tissue in muscles and repair segmental bone defects. SDBM is a kind of effective natural scaffold in tissue engineering of bone.

Animals ; Bone Demineralization Technique ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; Bone Matrix ; Implants, Experimental ; Male ; Rabbits ; Radius Fractures ; surgery ; Random Allocation ; Stem Cells ; cytology ; Tissue Engineering

Objective：This study was designed to explore the effect of BRD7 gene negative associated with nasopharyngeal carcinoma (NPC) on the growth of NPC cell line HNE1. Methods： The mammal expression vector of BRD7, pcDNA3.1(＋ )/BRD7, was constructed and transfected into HNE1 cell. Stable G418 resistant clones were isolated, and the integration of the exogenous vector DNA and the expression of BRD7 gene were detected by Southern blot and RT PCR respectively. Finally the cytobiological characterization of positive clone (B 4) was analyzed by using population double time (PDT), soft agar assay, cytometry, and xenograft. Results： The PDT of G418 resistant HNE1 cell with epression of BRD7 was 53 h and significantly longer than that of vector transfected HNE1 cell and untransfected HNE1 (P< 0.01). Flow cytometric data shown that more BRD7 transfected cells went into phase G0－ G1 than controls. And it also presented decreased clonogenicity and tomorigenicity in soft agar assay and tumor formation in nude mice. Conclusion： The reexpression of BRD7 could favor the malignant phenotype revision of NPC cells. And BRD7 gene might be a good candidate of tumor suppressor gene correlated with NPC.

Streptomyces coelicolor is the model species among streptomycetes. Until now, proteomic analyses of S. coelicolor have been conducted using two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry method, few integral membrane proteins were identified due to the hydrophobic and low-abundance nature of these proteins. In this work, 154 possible inner membrane proteins from S. coelicolor were identified using high pH-proteinase K sample preparation method and multidimensional protein identification technology, among them 44 are integral membrane proteins containing at least one transmembrane domain, most peptides and their corresponding proteins were identified experimentally for the first time.
Bacterial Proteins ; analysis ; Cell Membrane ; chemistry ; Genome, Bacterial ; genetics ; Mass Spectrometry ; methods ; Membrane Proteins ; analysis ; Proteome ; analysis ; genetics ; Streptomyces coelicolor ; chemistry ; genetics

Objective:To evaluate and compare the efficacy of laparoscopic and open tension-free surgery for adult unilateral primary inguinal hernia.Methods:Randomized controlled trial(RCT)reports published from 2006 to 2016 were searched in the PubMed,EMBASE,Web of Science,CBM and VIP databases.Data related to the clinical outcome,which compared the efficacy of laparoscopic and open surgery for adult unilateral primary inguinal hernia,were independently extracted and analyzed by two reviewers according to Cochrane Handbook for Systematic Reviews of Interventions 5.0.1.Statistical analysis was performed using RevMan 5.3 software.Objective:A total of 3 200 adult patients with unilateral primary inguinal hernia from 12 RCTs in 13 papers were enrolled in this Meta-analysis.Compared with open surgery,the cost of hospitalization in laparoscopic repair group increased(SMD=4.99,95％CI 3.13-6.85,P<0.001),and the rates of complication occurrence(RR=0.82,95％CI 0.68-0.98,P=0.03),NRS(MD=-1.67,95％CI-2.29——1.05,P<0.001)and persistent pain recurrence(RR=0.40,95％CI 0.23-0.69,P=0.001)reduced.However,there was no significant difference in the operation time(MD=0.14,95％CI 2.69-2.97,P=0.92)and the recurrence rate(RR=0.96,95％CI 0.32-2.88,P=0.06).Conclusions:Compared with open surgery,laparoscopic surgery is superior for adult primary unilateral inguinal hernia,but with higher cost.Accordingly,the appropriate surgical operation should be selected.

OBJECTIVETo study the expression and possible function of RhoA in human gastric cancer cell lines.

METHODSThe expression of RhoA in human gastrointestinal cancer cell lines was detected by Western blot. Antisense plasmid of RhoA was constructed by pGEFL and transferred into gastric cancer cell line AGS by lipofectamine. Cell survival was examined by MTT assays, and cell cycle was detected by flow cytometry.

RESULTSThe expression of RhoA protein in 10 different kinds of human cancer cell lines was much higher than that in immortalized human intestinal epithelial cell line. After being transfected with antisense RhoA, with the decrease in RhoA protein expression, the growth rate of AGS was inhibited, and the number of cells in S phase was increased by 14%.

CONCLUSIONRhoA is overexpressed in many human cancer cell lines. Some of the malignant characteristics of a gastric cancer cell line can be partially reversed by inhibiting RhoA expression.

Antisense Elements (Genetics) ; pharmacology ; Cell Cycle ; Cell Line, Tumor ; Genetic Therapy ; Humans ; Stomach Neoplasms ; chemistry ; pathology ; therapy ; rhoA GTP-Binding Protein ; analysis ; antagonists & inhibitors ; physiology

OBJECTIVETo identify genetic variation of HIV-1 predominant subtype B and C strains in China during rapid horizontal transmission and to elucidate the potential relationship between genetic variation and selective pressure.

METHODSAfter the fragments of HIV-1 env gene were amplified by nested-PCR from the whole blood of 258 HIV-1 infected individuals, PCR products were directly sequenced using ABI 377 DNA sequencer. The sequences covering env V3-V4 region of 72 HIV-1 subtype B(n=37) and C(n=35) strains were selected for phylogenetic analysis. In addition, the ratios of synonymous (Ks) substitutions per nonsynonymous (Ka) substitutions were calculated using DIVERGE.

RESULTSThe genetic distances of the V3-V4 region of subtype B strains were higher than that of subtype C strains. Furthermore, sequence analysis revealed that the V4 region was more variable than the V3 region for both subtype B and C strains. What's more, the V3 loop was less variable compared with the V3 upstream region and C3 region for subtype C Ks/Ka ratios of the entire aligned sequence of the two subtypes were below 1 0, with the lowest values found in the V3 region of subtype B strain and the V4 region of subtype C strain.

CONCLUSIONSThe majority of variation in both subtypes B and C strains occurred in the V4 rather than the V3 region. It is important that our study found for the first time the V3 loop was more conservative than the V3 upstream region and C3 region for subtype C. Calculations of the Ks/Ka ratios throughout the V3-V4 region demonstrate that significant selective pressures experienced during the rapid horizontal spread of the virus in the Chinese HIV-1 infected population may have directed change in the V3 loop for the subtype B strain and the V4 loop for the subtype C strain. These results will contribute to the policy of AIDS prevention and control and the ongoing development vaccine.

Amino Acid Sequence ; China ; epidemiology ; Gene Products, env ; genetics ; Genetic Variation ; HIV Infections ; epidemiology ; virology ; HIV-1 ; genetics ; Humans ; Molecular Sequence Data ; Phylogeny ; Sequence Analysis, DNA ; Sequence Analysis, Protein

OBJECTIVETo investigate the relationship between immunogenicity and decellularization processes of chemically acellular nerve allografts.

METHODSAdult Sprague Dawley rats were used as nerve donors and adult male Wistar rats used as nerve recipient hosts. 25 mm nerve segments were excised from SD rats' sciatic nerves. The nerve segments were decellularized via an improved chemical decelluarization treatment as follows: (1) nerve segments were rinsed with cold sterile Ringer's solution; (2) stabilized by pinning the ends to a thin plastic support, and submerged in 4% Triton-100 solution 12 h; (3) soaked into 3% sodium deoxycholate for 12 h; (4) washed in distilled water for 6 h. The procedures were repeated once again. The acellular nerve allografts from SD rats were sterilized by gamma irradiation and implanted into Wistar rats subcutanously. The control group was implantation of fresh nerve allografts from SD rats. The immunogenicity of acellular nerve allograft was tested by immunohistochemical examination of the intensity of CD3(+), CD4(+) and CD8(+) cells that infiltrated the allografts. Ulnar nerve segments were obtained from forearms of dogs and decellularized according to above procedures. According as the decellularization times, The ulnar nerve segments were divided into three subgroups: in group I, group II and group III, the nerve segments were decellularized repeatedly two, three and four cycles respectively. Each ulnar nerve segment was subdivided into five portions from proximal to distal end. The degrees of decellularization, demyelination and basal lamina integrity of extracellular matrix scaffold were observed with microscope and assessed by a score system. The immunohistochemical staining of GAG was observed.

RESULTSThe intensity of CD3(+), CD4(+) and CD8(+) T cells that infiltrated the allografts was greatly lower in acellular nerves than in fresh nerves. The mild cell-mediated host-graft immunorejection in acellular nerves was observed. On the decellularization procedures, the cells were completely extracted from nerves in all groups, but the myelin sheath were partially existed, and the GAG was present in the basal membrane of myelin sheath. In the score of demyelination, there were no statistical differences between groups (P > 0.05). The statistical difference of basal lamina integrity scores between group I and group II, group I and group III were significant (P < 0.05). As increasing the times of process, the degrees of disintegrity of basal lamina was significantly enhanced.

CONCLUSIONSAlthough decellularization processes significantly reduce the cell-mediated immunorejection of acellular nerve allografts, it can induce mild immunoreaction all the same, the antigen that responsible for immunogenicity may be the residual component of GAG in myelin sheath.

Animals ; Cell Separation ; methods ; Dogs ; Immunohistochemistry ; Male ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Sciatic Nerve ; cytology ; immunology ; transplantation ; Transplantation, Homologous ; immunology ; Ulnar Nerve ; anatomy & histology ; cytology