Action Potentials ; drug effects ; physiology ; Animals ; Female ; Guinea Pigs ; Heart Ventricles ; drug effects ; Papillary Muscles ; drug effects ; physiology ; Progesterone ; pharmacology

Background Some drugs with inhibitory effect on the proliferation of lens epithelial cells have a limiting application in clinic because of their adverse response.To screen the effective and less side-effect drug for supressing LECs growth is very inportant for the prevention and treatment of after cataract.Objective This study was to explore the effects of docetaxel on LECs growth and compare its role with epirubicin hydrochloride,pirarubicin hydrochloTide and rahitrexed.Methotis Immortalized human LECs line (SRA01/04) were cultured and passaged.Different concentrations of docetaxel,epirubicin hydrochloride,pirarubicin hydrochloride and rahitrexed were added into the medium respectively for 24.48 and 72 hours.The proliferation of LECs was detect by M1Yr.Flow cytometry analysis Was used to analyze the influence of different concentrations of docetaxel on cellular cycle at 48 hours after addition of docetaxel,and Annexin V-FITC/PI marking method was used to assesse the apoptosis of LECs under the action of docetaxel.Expression of bcl-2 protein in LECs Was evaluated by Westeru blot. Result The growth rate of LECs Wag 100%in 8-519 pmol/L doeetaxel groups with the normal cell shape.Majority of abnormal cells and low growth rate were found in 66 nmoVL docetaxel group at 48 and 72 hours.The IC50 of docetaxel was lowest in 48 and 72 hours in docetaxel group in comparison to epirubicin hydrochloride and pirarubicin hydrochloride. However,no evident inhibition on LECs growth in 23.22-523.56 μmol/L of raltitrexed.At 48 hours,the percentage of LECs in G2/M phase increased as the asccnte of concentration of docetaxel,showing a significant difference among 4 groups(F=2633.05,P＜0.01).The percentage of early apoptotic cells increased to 22.4%(χ2=20.00,P＜0.01) and 27.9%(χ2=42.68,P＜0.01)from normal control 3.1% at 48 hours after LECs exposed to 8.3 nmol/L and 266 nmol/L docetaxe.The expression of bcl-2 protein in LECs was obviously weakened after addition of docetaxel,especially 8.3 nmol/L docetaxel group. Conclusion Docetaxel,epirubicin hydrochloride and pirarubicin hydrochloride can inhibit the proliferation of human LECs in vitro.But there is no supression on LECs growth inraltitrexed.Docetaxel is proved to have a strongly arrested effect on the proliferation of LECs in comparison with epirubicin hydrochloride and pirarubicin hydrochloride and play its role at concentration-and time-dependent manner.

Objective To explore the changes of CD40 and CD40 ligand(CD40L) levels and investigate their significances in children with graft-versus-host disease(GVHD)after related-donor human leukocyte antigen(HLA) matching allogeneic hematopoietic stem cell transplantation(HSCT).Methods Nineteen patients with ?-thalassemia major and 1 patient with congenital inherent hemolytic anemia accepted umbilical cord blood transplantation(UCBT) and allogeneic peripheral blood stem cell transplantation(allo-PBSCT),respectively,and all cases were received successfully related-donor human leukocyte antigen matching allogeneic HSCT.Peripheral blood samples were obtained before and after transplantation,the time when GVHD happened,the expressions of CD40 and CD40L were measured by using immunofluresence asssy.Results Four UCBT children and 3 allo-PBSCT children had no acute GVHD.Thirteen children had acute GVHD(degreeⅠ-Ⅳ),the expressions of CD40L on CD4+ and CD8+ T cells in the patients with acute GVHD increased,especially in allo-PBSCT.Five cases of UCBT and 12 cases of allo-PBSCT patients had chronic GVHD,the expressions of CD40L+,CD25+ and CD69+ on CD4+ and CD8+ T cells in patients with chronic GVHD increased obviously.The expression of CD19+CD40+ was lower than normal within 3 months after transplantation.Conclusions The high expression of CD40L+T cells in periferal blood after HSCT was related to the activation and proliferation of T cells in the development of GVHD in HSCT.

Objective To construct cDNA entry library and cDNA expression library of Armillifer agkistrodontis (A.) nymphs and make a preliminary immunoscreening for the cDNA expression library.Methods The nymphs were collected from the Kunming mice infected experimentally with A.agkistrodontis eggs and the total RNA were extracted from the nymphs using TRIzol Reagent.After purifying the mRNA,the synthesized cDNAs were cloned into the donor vector pDONR222 by BP reaction of Gateway technology and the recombinants were transformed into the DH10B cells by electroporation,the cDNA entry library was obtained.Next,the expression vector pDEST17 was ligated with entry clones by LR reaction,and the recombinants were transformed into the BL21 (DE3) cells.Hence,the cDNA expression library was constructed.Then,the expression library was immunoscreened with the mixed sera of mice infected with A.agkistrodontis,and the insertions of positive clones were sequenced.After that,the open reading frame(ORF) of positive slone sequence,the homology of the screened genes and their encoded proteins were analyzed by Finder and BLAST (basic local alignment search tool) program of National Center of Biotechnology Information(NCBI),and the discovered new genes were submitted into the GenBank.Besides,the physico-chemical properties,secondary structure and B cell epitopes of encoded proteins were also analyzed by bioinformatics software.Results The average titer and total clones of the cDNA entry library were 1.45 × 105 CFU/ml(colony-forming unit,CFU) and 1.74 × 106 CFU,respectively,and the range of fragment length of the inserted cDNA was between 0.2-4.0 kb,with an average of 1.4 kb.The total clones of cDNA expression library were 1.00 × 105 CFU,and the fragment length of the inserted cDNA was between 0.3-2.2 kb,with an average of 1.0 kb.Five positive clones,coded S1,S5,A1,D1 and F1,respectively,were obtained through preliminary immunoscreening.The sequence and homology of the five positive clones were sequenced and analyzed by BLAST program.No significant similarities were found in pentastomida species,which meant that they were all novel genes of A.agkistrodontis.The gene sequences were submitted to GenBank,with the accession number from JQ180451 to JQ180455.Also,results obtained by bioinformatics software showed that the predictive encoding proteins were all potential to be valuable recombinant diagnostic antigens.Conclusions The cDNA library of A.agkistrodontis nymphs is successfully constructed,and five new genes of A.agkistrodontis are discovered.The establishment of cDNA library and the discovery of the new genes will lay a foundation for further studying the gene functions and screening the immunodiagnostic antigens.

In order to obtain higher L-lactic acid yield industrial strain, the original strain Rhizopus oryzae PW352 was mutated by means of N+ ions implantation and a mutant strain Rhizopus oryzae RE3303 was obtained. Its lactic acid yield was increased by 75% than that of the original one. The acid producing condition was optimized by orthogonal design. The concentration of L-lactic acid reached to 131～136g/L and the conversion rate of glucose was as high as 86%～90% under the optimum condition.

Objective To explore the expression and distribution of calcineurin(CaN)in normal and failing human myocardium.Methods Left and right ventricles were obtained from end-staged heart failure patients(n=12)undergoing heart transplantation and donor hearts(n=5)taken from victims of vehicle accidents.Immunohistochemistry and SDS-PAGE technique were used to demonstrate expression and distribution of CaN. Results Positive immunoreactive staining for CaN was detected in human cardiomyocytes, cardiac fibroblasts and epicardial mesothelial cells,but not detected in cardiac vascular endothelial cells and smooth muscle cells.There was no difference in CaN protein levels between failing hearts and donor hearts (Band intensity of right ventricle in failing hearts and donor hearts was 130.20±8.66 and 139.87±6.21, P=0.33.Band intensity of left ventricle in failing hearts and donor hearts was 106.45 and 126.34 ±12.09)and between left ventricular and right ventricular myocardium(Band intensity of left and right ventricles in failing hearts was 96.99±10.67 and 104.58±13.18, P=0.63.Band intensity of left and right ventricles in failing hearts was 132.12 and 120.74).Conclusions CaN is expressed in human cardiomyocytes, fibroblasts and epicardial mesothelial cells and the protein level and distribution of CaN are similar in failing and donor hearts.

Açaí (Euterpe oleracea) emerged as a source of herb has a long history in South America, which was approved by the Ministry of Health used in China and it has been introduced planting in Guangdong and Taiwan. This article summarized applied history of Açaí and its present status in China. Did theoretical study on the Chinese herbal properties of Açaí based on the Chinese traditional philosophical culture to analysis the function and symptom preliminary, combining with used for medical recordation, chemical component, biological activity. It is aiming at establishing the theoretical foundation for the application under the guidance of TCM theory.
Euterpe ; chemistry ; Medicine, Chinese Traditional ; Models, Theoretical ; Phytotherapy ; Plant Extracts ; pharmacology ; South America

OBJECTIVETo investigate the feasibility of chondrogenesis in vitro with bone marrow stromal cells (BMSCs) induced by the co-cultured chondrocytes.

METHODSThe BMSCs and chondrocytes were separated from pig and cultured. The supernatant of chondrocytes was used as the inducing solution for BMSCs from the 2nd generation. 7 days later, samples were taken and underwent immunohistochemistry and RT-PCR for detection of the expression of specific type II cartilage collagen, type II collagen and aggrecan mRNA. The cultured BMSCs and chondrocytes were mixed at a ratio of 8:2 (BMSC: cartilage cell) and were inoculated into a polyglycolic acid/polylactic acid (PGA/PLA) scaffold at the final concentration of 5.0 x 10(7)/ml. The cartilage cells and BMSCs were also inoculated separately at the same concentration as the positive and negative control. Pure cartilage cells at 20% of the above mentioned concentration (1.0 x 10(7)/ml) were used as the low concentration cartilage cell control group. Samples were collected 8 weeks later. General observations, wet weight, glycosaminoglycans (GAGs) determination and histological and immunohistochemistry examinations were performed.

RESULTSThe expression of type II collagen, type II collagen and aggrecan mRNA were positive in induced BMSCs. In the co-cultured group and the positive control group, pure mature cartilage was formed after 8 weeks of culture in vitro, and the size and shape of the scaffold were maintained. The newly formed cartilage in the two groups were almost the same in appearance and histological properties. The immunohistochemistry results indicated that the cartilage cells of the two groups all expressed ample cartilage-specific type II collagen. The average wet weight and GAG content in the co-cultured group reached more than 70% of those in positive control group. Only an extremely small amount of immature cartilage tissues was formed in local regions in pure BMSC group, and the scaffold was obviously shrunk and deformed. Although the wet weight of newly generated cartilage tissue in the low concentration cartilage cell group reached 30% of that in positive control group, the scaffold was obviously shrunken and deformed. Only regional and discontinuous cartilage tissues were formed, and the amount of newly formed cartilage was obviously less than that in the co-culture group and the positive control group.

CONCLUSIONSChondrocytes can provide a micro-environment for the formation of cartilage, and also effectively induce BMSC to differentiate into chondrocytes and form tissue-engineered cartilage in vitro.

Aggrecans ; metabolism ; Animals ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; Coculture Techniques ; Collagen Type II ; metabolism ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Swine ; Tissue Scaffolds

OBJECTIVETo establish chemical pattern recognition method for the identification and evaluation of Atractylodes macrocephala.

METHODThe chemical constituents in methanol extract of 32 samples of A. macrocephala were determined by HPLC. The fingerprints were obtained and were handled by hierarchical clustering analysis and stepwise discriminant analysis.

RESULT AND CONCLUSIONAccording to the result of classification, all samples collected were devided into three Grades--the superior, the ordinary and the fake. Chemical pattern recognition method was established. It may be of practical value for the quality control of A. macrocephala.

Atractylodes ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Cluster Analysis ; Pattern Recognition, Automated ; Plants, Medicinal ; chemistry ; Quality Control ; Rhizome ; chemistry

BACKGROUND AND OBJECTIVEEpstein-Barr virus (EBV) has been detected in about 10% of gastric carcinomas. However, the pathogenetic role of EBV in gastric carcinoma is uncertain. This study was to explore the correlation of Fas/FasL expression to the apoptosis of tumor cells and tumor-infiltrating lymphocytes (TIL) in EBV-associated gastric carcinoma (EBVaGC).

METHODSFas/FasL expression in 49 specimens of EBVaGC, 20 specimens of EBV-negative gastric carcinoma (EBVnGC) and 12 specimens of normal gastric mucosa was detected by immunohistochemistry. The apoptotic index (AI) of cells was determined by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL).

RESULTSThe positive rates of Fas were 91.7% in normal gastric mucosa and 76.8% in gastric carcinoma (P < 0.05); those of FasL were 16.7% in normal gastric mucosa and 58% in gastric carcinoma (P < 0.05). The positive rate of Fas was significantly lower in EBVaGC than in EBVnGC (71.4% vs. 90.0%, P < 0.05). The positive rate of FasL in EBVaGC was significantly higher than that in EBVnGC (63.2% vs. 45%, P < 0.05). The AI of EBVaGC cells was significantly lower than that of EBVnGC cells (P = 0.002). The number and AI of TIL in EBVaGC were higher than those in EBVnGC (P < 0.05). The AI of TIL was positively correlated with the level of FasL expression in tumor cells (r=0.237, P = 0.028).

CONCLUSIONUp-regulation of FasL expression and decrease of TIL apoptosis in EBVaGC may facilitate the escape of tumor cells from the host immunosurveillance, and it might contribute to the development and progression of carcinoma.

Adult ; Apoptosis ; Epstein-Barr Virus Infections ; Fas Ligand Protein ; metabolism ; Female ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Immunohistochemistry ; Immunologic Surveillance ; In Situ Nick-End Labeling ; Lymphocytes, Tumor-Infiltrating ; pathology ; Male ; Middle Aged ; Stomach Neoplasms ; metabolism ; pathology ; virology ; Tumor Escape ; fas Receptor ; metabolism