OBJECTIVE: To study the preparation technology of tamsulosion hydrochloride sustained-release capsules and investigate the release degree in vitro. METHODS: The pellets containing tamsulosin hydrochloride were prepeared in the fluid-bed using bottom gush medicine. Then, it was coated with ethylcellulose aqueous dispersion (surelease), and in the following procedures, water-based acrylic resin enteric system (Acryl-EZE®) was used as coating material, hydroxypropylmethylcellulose E6 (HPMCE6) was considered as porous agent by fluid-bed. Based on the release degree in vitro, prescription influence factors were evaluated, as well as drug releases curve was compared, according to the single factor experiment. RESULTS: The preparation technology referred in our research was available to make tamsulosin hydrochloride sustained-release capsules, and drug release curve of self-made sustained-release capsules was similar to the commercial one. Additionally, the products reproducibility of intra-batch and inter-batch was excellent. CONCLUSION: The tamsulosin hydrochloride sustained-release capsules prepared in this study exhibited ideal sustained-release characteristics in vitro. The formulation is reasonable and feasible. It is suitable for industrial production.

BACKGROUND:Chitosanase is an enzyme for efficient and special hydrolysis of chitoan, and hence its effective and stable expression with enzymatic activity wil contribute to improving gene therapeutic effect.
OBJECTIVE: To construct a chitosanase gene for the efficient and specifical hydrolysis of chitosan, and to investigate its expression inEscherichia coli and the main influencing factors of enzymatic activity.
METHODS:According to the sequences of endo-chitosanase ofAspergilus sp. CJ22-326 provided in Genbank (EU302818), primers were designed and synthesized. The Asperguilus endo-chitosanase gene was amplified by successive extension PCR. And then the recombinant pET28a-His6-CSN was constructed and expressed in
Escherichia coli BL21. Finaly the recombinant His6-CSN fusion protein was analyzed by sodium dodecyl sulfate polyacrylamide gelelectrophoresis (SDS-PAGE), the western blot and dinitrosalicylic acid assay for detecting the enzyme activity of eluted His6-CSN fusion protein. The influence of different pH value and temperature on the enzyme activity of the recombinant chitosanase was investigated.
RESULTS AND CONCLUSION: SDS-PAGE showed that 29 kDa proteins were expressed and the western blot assay showed that His6-CSN expressed successfuly in the host. Dinitrosalicylic acid assay determined the
enzymatic activity of His6-CSN was significantly higher than that of lysozyme, but lower than that of chitosanase from Streptomyces griseus (P < 0.05). The recombinant chitosanase displayed the maximal activity at temperature of 50℃ and pH value of 6.0. There were a higher enzymatic activity remaining at pH value of 4.0-7.0 and
temperature of 30-50℃.

Objective:To analyze the correlation of single nucleotide polymorphisms ( SNP) of ATP binding cassette transporter A1 gene (ABCA1) and lower extremity atherosclerotic disease (LEAD). Methods: The clinical data and peripheral blood were col-lected from 630 participants (314 LEAD cases and 316 normal controls) in Han population of Minnan. The 9 SNP genotypes in the ABCA1 gene were detected by Sequenom MassArray. Results:Among the 9 SNP genotypes, rs2980083 was rejected because it wasn' t in accordance with Hardy-Weinberg equilibrium. Obvious linkage disequilibrium was found between rs2066714 and rs2066715, rs1800976 and rs2246293, rs2246293 and rs2980083, and rs1800976 and rs2980083(D′>0. 9,r2 >1/3). There were no significant differences (P>0. 05) in 6 haplotypes of ABCA1 gene groups between the LEAD cases and the normal controls. No significant differ-ences (P>0. 05) were found in frequency distribution between the LEAD cases and the normal controls in 8 SNP according to the re-sults of genotype statistics. There was no onset risk of LEAD according to the gene logistic regression analysis. Conclusion:The SNPs of rs10124755, rs2980083, rs1800976, rs4149341, rs2066714, rs2066715, rs2066716, rs2230808 and rs2246293 might not correlate with the susceptibility of LEAD in Han population of Minnan.

BACKGROUND:Sodium alginate and chitosan are the polycation and polyanion natural polymer materials respectively, and they can be crosslinking agents complementing each other to form composite gel and avoid the cytotoxicity resulting from some common crosslinking agents .
OBJECTIVE:To prepare the chitosan-sodium alginate composite gel and evaluate its cytotoxicity in vitro. METHODS:Chitosan was dissolved in 0.25 mol/L acetic acid to make a 30 g/L mass concentration solution, and 0.1 mol/L NaOH solution was added to neutralize its acidity. Neutralization of the chitosan solutions leads to the formation of a precipitate in ultrasmal particles. Then the chitosan and 3%sodium alginate solution in deionized water were mixed in 1:1 volume ratio by high frequency oscil ating to produce composite gel. The composite gel were detected by scanning electron microscopy and Fourier transform infrared spectrometry after freeze-drying. The 24-hour and 72-hour leaching solutions of composite gel, 24-hour and 72-hour leaching solutions of polyethylene and phenol solution were added to the L-929 cells’ culture medium respectively in order to evaluate the cytotoxicity of composite gel in vitro.
RESULTS AND CONCLUSION:The results of Fourier transform infrared spectrometry showed the variation of characteristic peak values of composite gel which were different from sodium alginate and chitosan;and under scanning electron microscope, a spatial network structure formed with abundant intervals. Result of the cytotoxicity valuation was qualified for the chitosan-sodium alginate composite gel. These findings indicate that the chitosan-sodium alginate composite gel can be used as tissue engineering scaffold materials.

BACKGROUND:Chitosan is wel known as good biocompatibility and biodegradability;however, its extensive use in biomedical applications is restricted due to its poor transfection efficiency. OBJECTIVE:To prepare the polyethyleneimine-chitosan/DNA nanoparticles loading enhanced green fluorescent protein gene, and to detect their physicochemical properties and gene transfection efficiency towards chondrocytes in vitro.
METHODS:Low molecular weight polyethyleneimine was covalently linked to chitosan backbone to construct chitosan-graft-polyethyleneimine;then the chitosan-graft-polyethyleneimine was mixed with DNA nanoparticles, which loaded enhanced green fluorescent protein gene, by a complex coacervation method. The nanoparticle morphology was observed under a scanning electron microscopy. The sizes and zeta-potentials of the
nanoparticles were measured by a Marven-nano laser diffractometer. The binding capacity of plasmid DNA was evaluated by agarose gel electrophoresis analysis. The gene transfection experiments in vitro were performed towards rabbit’s chondrocytes. The gene transfection efficiency was measured with flow cytometry and under fluorescence microscope. How marked DNA entered into the nucleus of chondrocytes mediated by the nanoparticles was detected by laser scanning confocal microscopy.
RESULTS AND CONCLUSION:The prepared nanoparticles were mainly spherical, with an average size of (154.6±18.6) nm, and zeta-potential of (24.68±6.82) mV. The agarose gel electrophoresis analysis confirmed that the nanoparticles could effectively protect plasmid DNA from DNase Ⅰ-induced degradation. Gene transfection in vitro proved that the nanoparticles were efficient in transfecting rabbit’s chondrocytes and the expression of green fluorescent proteins was observed under fluorescent microscope, with a transfection efficiency of (23.80±1.74)%that was significantly higher than that of the naked plasmid DNA and chitosan/DNA nanoparticles (P<0.05). But no significant differences were observed between polyethyleneimine-chitosan/DNA nanoparticles and LipofectamineTM 2000. These findings indicate that the polyethyleneimine-chitosan/DNA nanoparticles can effectively protect plasmid DNA from nuclease degradation, and exhibit the favorable transfection ability towards articular chondrocytes.

Congenital Hyperinsulinism ; diagnosis ; Humans ; Infant, Newborn ; Male

OBJECTIVE: The significance of lymph node dissection in the VI area of cN0 thyroid papillary carcinoma. METHOD: Collect 150 cases of patients diagnosed with cNO thyroid papillary carcinoma and they were performed thyroid gland lobe and isthmic portion excision including lateral VI area lymph node cleaning. The specimens were pathologic examined to determinate the size, the position, invasion of thyroid papillary carcinoma,the number and metastasis of lymph node, etc. RESULT: In the 150 patients performed the lymph node VI area groups cleaning, 93 cases had VI area of lymph node metastases, so the transfer rate was 62.0%. In the VI area, metastasis rate of tracheal side lymph nodes was 62.0% (93/150), lymph node before throat group was 4.67% (7/150), lymph node before trachea group was 3.33% (5/150), lymph nodes near the trachea laryngeal recurrent nerve ventral group was 52.0% (78/150), and next to the trachea laryngeal recurrent nerve dorsal lymph node group was 21.33% (32/ 150). CONCLUSION In CN0 thyroid papillary carcinoma, VI zone of lymph node metastasis rate is high, and region VI lymph node metastasis rate from high to low in order for: paratracheal lymph node, prelaryngeal lymph node, pretracheal lymph node. The metastasis rate of paratracheal throat back nerve ventral lymph node was the highest in central lymph node.
Carcinoma ; pathology ; surgery ; Carcinoma, Papillary ; pathology ; surgery ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Neck ; Neck Dissection ; Recurrent Laryngeal Nerve ; Thyroid Cancer, Papillary ; Thyroid Neoplasms ; pathology ; surgery

Objective To express and detect the antigenicity of human laminin alpha4 LG3-4 module (hLN?4LG3-4) protein by gene engineering techniques. Methods The cDNA encoding hLN?4LG3-4 was amplified by RT-PCR from human placenta, then inserted into pMD-18T vector by T/A cloning and sequenced. Prokaryotic expression vector pET-28a-LG3-4 was constructed by recombinant DNA technique. The hLN?4LG3-4 fusion protein expressed in BL21(DE3)/pET system was identified by SDS-PAGE, purified by Ni-NTA resin, and assayed by Western blotting. Results The cDNA fragment of hLN?4LG3-4 was cloned successfully. While BL21 (DE3)/pET28a-LG3-4 bacterium was induced with IPTG, a new protein band with a relative molecular weight of 44 000 was shown on SDS-PAGE profile. hLN?4LG3-4 fusion protein of high purity (95%) was obtained and specific protein band was detected by Western blotting. Conclusion The hLN?4LG3-4 fusion protein was successfully expressed.

Objective: To identify and preliminarily analyze the plants of Marsdenia R. Br. and its related species by ITS2 and rbcL sequences. Methods: The ITS2 and rbcL regions were amplified and sequenced directly. The sequence was manually calibrated using CodonCode Aligner, and splited and jointed by Contig software. The genetic distance analysis was carried out by MEGA 4.0 and the phylogenetic tree was constructed using the Neighbor-joining method. Results: The samples analyzed were well identified by ITS2 and rbcL sequences, and the authentication ability of ITS2 was superior to rbcL. The genetic relationship among the plants of Marsdenia R. Br., Dregea E. Mey., and Gymnema R. Br. was very close. Conclusion: ITS2 and rbcL sequences could distinguish Marsdenia R. Br. species very well, and the authentication ability of ITS2 is superior to rbcL. The preliminary studies suggest that the Dregea E. Mey. and Gymnema R. Br. should be classified into genus Marsdenia R. Br.

AIM: To construct the prokaryotic expression system containing protein transduction domain (PTD) with heat shock protein 27 (HSP27) in order to prepare and purify the recombinant protein , and to verify whether the recombinant protein PTD-HSP27 has the ability to penetrate the human lens epithelial cell ( HLEC) membrane and the rabbit cornea.METHODS:The plasmid pKYB-PTD-HSPB1-6His was constructed by the technique of overlap extension PCR.The plasmid was transformed and PTD-HSP27 was purified through nickel affinity chromatography column and identi-fied by Western blotting.PTD-HSP27-6His was labeled with the fluorescein isothiocyanate (FITC).The penetrating ability of PTD-HSP27 into HLECs and rabbit cornea was tested .RESULTS:The recombinant PTD-HSP27 plasmid was success-fully cloned and effectively expressed .The correctness of the recombinant protein PTD-HSP27 was demonstrated .Fluores-cence microscopic examination showed that PTD-HSP27-FITC was internalized by HLECs .Fluorescent labeled PTD-HSP27 was then observed in the rabbit aqueous humor .CONCLUSION:The recombined gene PTD-HSPB1 was constructed by o-verlap extension PCR technique and the PTD-HSP27 fusion protein was prepared and purified by nickel affinity chromatog-raphy column.Using the technique of PTD-fusion protein, HSP27 was transduced into HLECs and passed through the cor-nea .