Objective:To produce rhBMP-4 with bioactivity in E.coli. Methods: The full-length human BMP-4 gene was mutated by PCR without changes in amino acid sequence, then the synthesized gene was cloned into plasmid pET-3c, transducted into BL21(DE)plysS, and induced by adding IPTG to a final concentration of 1.0 mmol/L. The protein product was purified using ion-exchange chromatography method and then renaturated, bioactivity was checked by C2C12 differentiation in vitro and mouse ectopic bone formation in vivo. Results: A 438 bp gene fragment encoding mature peptide of hBMP-4 was cloned , the protein product was mostly in the form of inclusion body, after renaturation, the engineering protein shows better bioactivity. Conclusion:The mutant strategy can enhance the expression of bioactive rhBMP-4 in E.coli expression system.

OBJECTIVETo compare efficacy of different adjuvant chemotherapy regimens for stage II-III gastric cancer after D2 gastrectomy in Asian patients.

METHODSAssociated literatures were searched through electronic databases and hand-searching. Prospective randomized clinical trials (RCTs) comparing adjuvant chemotherapy after D2 gastrectomy with surgery alone were included in the study. Overall survival and disease-free survival were chosen as the endpoints. Relative hazard was analyzed by Bucher adjusted indirect comparison.

RESULTSTwo RCTs were selected, including comparison between S-1 versus surgery alone and comparison between XELOX versus surgery alone. There was no statistical difference in overall survival between the two regimens (HR=0.94, 95%CI:0.62-1.44, P=0.79). The recurrence risk of S-1 was slightly higher as compared to XELOX, but no statistical difference was found (HR=1.11, 95%CI:0.80-1.53, P=0.54).

CONCLUSIONThe adjuvant chemotherapy with S-1 is similar to XELOX for stage II-III gastric cancer after D2 gastrectomy in Asian patients.

Antineoplastic Combined Chemotherapy Protocols ; Chemotherapy, Adjuvant ; Deoxycytidine ; analogs & derivatives ; Fluorouracil ; analogs & derivatives ; Humans ; Postoperative Care ; Randomized Controlled Trials as Topic ; Stomach Neoplasms ; drug therapy ; surgery ; Treatment Outcome

Animals ; Fatty Liver ; drug therapy ; metabolism ; prevention & control ; Liver ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism ; Taurine ; therapeutic use

OBJECTIVETo explore the expression and significance of secretions of hypothalamus-pituitary-adrenal (HPA) axis in human hypertrophic scar.

METHODSHypertrophic scar tissues obtained from 12 patients with deep-partial thickness burn or full-thickness burn and normal skin tissues from the same 7 patients with hypertrophic scar were harvested for determination of gene expression of corticotrophin-releasing hormone (CRH), CRH receptor 1 (CRH-R1), pro-opiomelanocortin (POMC), melanocortin receptor 2 (MC-2R), and glucocorticoid receptor α (GR-α) by real-time fluorescence quantitative PCR. After addition of corresponding antibodies, distribution differences of CRH, CRH-R1, adrenocorticotropic hormone (ATCH), MC-2R, and GR-α were observed with immunohistochemical staining. Data were processed with t test.

RESULTSThe mRNA expression of CRH, CRH-R1, POMC, and GR-α in hypertrophic scar was respectively 3.1 ± 0.8, 0.05 ± 0.03, 0.020 ± 0.007, and 0.0030 ± 0.0010, which were significantly lower than those in normal skin (20.6 ± 4.7, 0.30 ± 0.12, 0.060 ± 0.020, and 0.0200 ± 0.0070, with t values from 2.10 to 4.75, P values all below 0.05). There was no statistical difference in MC-2R mRNA expression between hypertrophic scar and normal skin (t = 1.48, P = 0.15). Immunohistochemical observation showed CRH, CRH-R1, ACTH, MC-2R, and GR-α in hypertrophic scar were located in basal layer of epidermis, fibroblast of dermis, and tube wall of sweat gland. Expressions of these indexes could also be observed in sebaceous gland and hair follicle besides above-mentioned structures.

CONCLUSIONSDecreasing expression of active material of HPA axis may be related to formation of hypertrophic scar.

Adolescent ; Adrenocorticotropic Hormone ; metabolism ; Adult ; Child ; Cicatrix, Hypertrophic ; metabolism ; Female ; Glucocorticoids ; metabolism ; Humans ; Hypothalamo-Hypophyseal System ; metabolism ; Male ; Pituitary-Adrenal System ; metabolism ; Young Adult

OBJECTIVETo study the effect of melatonin on proliferation and apoptosis of fibroblasts in human hypertrophic scar and its mechanism.

METHODSFibroblasts from human hypertrophic scar were isolated and cultured with DMEM medium containing 10% FBS, and then they were divided into control (C, added with ethanol), low concentration (LC, added with 1 × 10(-5) mmol/L melatonin), middle concentration (MC, added with 1 × 10(-3) mmol/L melatonin), and high concentration (HC, added with 1 mmol/L melatonin) groups according to the random number table. After being cultured for 24 hours, cell morphologic change was observed under microscope; XTT-PMS assay was used to examine cell proliferative activity; cell cycle and apoptosis were assessed with flow cytometry after double staining of FITC and PI, and the levels of cyclin E, p53, and Fas mRNA were determined with fluorescence quantitative RT-PCR. Data were processed with analysis of variance and LSD test.

RESULTS(1) Fibroblasts in C group were spindle-shaped with growth in colonies. Along with the increase in melatonin concentration, fibroblasts in LC, MC, and HC groups gradually dispersed, deformed and atrophied, with shrunk cellular membrane, and decrease in ratio of nucleus and cytoplasm. (2) Proliferative activity of fibroblasts in LC, MC, and HC groups decreased along with an increase in melatonin concentration (1.49 ± 0.15, 1.24 ± 0.20, and 0.92 ± 0.09), which were lower that in C group (1.79 ± 0.10, F = 67.61, P < 0.05). Cell ratios of S and G2/M phases in LC, MC, and HC groups decreased along with an increase in melatonin concentration, which were all lower than those in C group [(10.6 ± 1.1)%, (6.1 ± 1.2)%, (3.2 ± 0.8)% vs.(16.9 ± 1.3)%, F = 286.10, P < 0.05; (13.5 ± 1.1)%, (9.8 ± 1.0)%, (6.0 ± 0.7)% vs. (16.7 ± 1.6)%, F = 162.69, P < 0.05]. Apoptotic rates in early and late stages of LC, MC, and HC groups increased along with an increase in melatonin concentration, all higher than those in C group (with F value respectively 424.05, 236.44, P values all below 0.05). The expressions of cyclin E mRNA in LC, MC, and HC groups decreased along with an increase in melatonin concentration, which were lower than that in C group (1.58 ± 0.21, 0.90 ± 0.20, and 0.24 ± 0.12 vs. 2.90 ± 0.30, F = 266.79, P < 0.05), while the expressions of p53 mRNA and Fas mRNA showed opposite tendency (with F value respectively 10.11, 12.03, P values all below 0.05).

CONCLUSIONSMelatonin can inhibit proliferation and induce apoptosis of fibroblasts in hypertrophic scar through regulating the gene expressions of cyclin E, p53, and Fas.

Adult ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Cyclin E ; metabolism ; Female ; Fibroblasts ; drug effects ; metabolism ; pathology ; Humans ; Male ; Melatonin ; pharmacology ; Oncogene Proteins ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; fas Receptor ; metabolism

OBJECTIVETo evaluate the release in fixed position of pH-dependent and enzyme-dependent Kangfuxin colon targeting capsules in vivo and in vitro.

METHODThe dissolution was tested in vitro and X-ray radiography was used for the evaluation in vivo.

RESULTAfter two hours pH-dependent colon targeting in man-made colon fluid, medicine release in fixed position on the whole, colon loc-release. Add enzyme into man-made colon, when enzyme-dependent colon targeting in it, then medicine release quickly, mainly release in fixed position; The conveying time in vivo of pH-dependent and enzyme-dependent capsules have big individuality difference. In the experiment, disintegration is stabilize among individuales, between 2.0-3.5 hours.

CONCLUSIONKangfuxin colon targeting capsules of two principles all release in fixed position to achieve the goal.

Animals ; Capsules ; Colon ; diagnostic imaging ; metabolism ; Delayed-Action Preparations ; chemistry ; Drug Carriers ; Drug Delivery Systems ; Female ; Humans ; Hydrogen-Ion Concentration ; Male ; Materia Medica ; administration & dosage ; isolation & purification ; pharmacokinetics ; Periplaneta ; chemistry ; Polygalacturonase ; chemistry ; Radiography

OBJECTIVETo prepare coated micro-pellets of pH-dependent and enzyme-dependent kangfuxin colon targeting delivery system, to make them go to colon, then release, educe partial effect.

METHODWe eploy pan-pill to prepare simple pellets, and prepare tunicatus pellets with fluidized bed coating. We investigated the preparation and parameter of pellets, so, we bolting the best shaping and tunicatus artwork.

RESULTThe ingredients for preparing the micro-pellets are 125% starch +2% CMC-Na, and add 30% ethanol to be binder, pellets were coated with Eudragit S100 to prepare ph-dependent and pectin-HPMC to prepare enzyme-dependent colon targeting micro-pellets.

CONCLUSIONWe get two micro-pellets of pH-dependent and enzyme-dependent kangfuxin colon targeting.

Animals ; Colon ; metabolism ; Delayed-Action Preparations ; Drug Carriers ; Drug Compounding ; methods ; Drug Delivery Systems ; Hydrogen-Ion Concentration ; Hypromellose Derivatives ; Materia Medica ; administration & dosage ; isolation & purification ; metabolism ; Methylcellulose ; analogs & derivatives ; Pectins ; Periplaneta ; chemistry ; Polymethacrylic Acids

OBJECTIVETo establish a bioluminescent MDA-MB-231 cell line which can stably express luciferase and green fluorescent protein to allow bioluminescent imaging in nude mouse models bearing human triple-negative breast cancer xenografts.

METHODSThe lentivirus carrying luc2, eGFP and neo fusion genes were packaged in 293T cells via calcium phosphate co-precipitation. Human triple-negative breast cancer cell line MDA-MB-231 was infected by the lentivirus, and the positive cell clones were tested for eGFP and luc2 expressions by fluorescence microscopy and Xenogen IVIS200 bioluminescent imaging system, respectively. MTT assay, transwell invasion assay and wound healing assay were performed to evaluate the changes in the proliferation, invasion and migration abilities of the infected cells. The cells were then orthotopically implanted into the right second mammary fat pat of female BALB/c nude mice. The tumor growth was monitored by the in vivo imaging system every week, and the tumor tissues were harvested to evaluate the in vivo stability and tumorigenicity of the modified cells using cryosection and HE staining.

RESULTSThe lentivirus-infected MDA-MB-231cells could stably express luc2 and eGFP, and the luciferase activity reached 9689 phontons/s/per cell. No significant changes occurred in the biological activities of the lentivirus-infected MDA-MB-231 cells. We successfully established the nude mouse model bearing orthotopically implanted human triple-negative breast cancer cells.

CONCLUSIONThe modified MDA-MB-231 cell line can be detected sensitively at the primary implantation site and distant metastasis site in nude mice, which provides a convenient and sensitive platform for the research of metastatic mechanism and new antitumor drugs of human triple-negative breast cancer. The combination of eGFP and luc2 is superior to single reporter gene.

Animals ; Brain Neoplasms ; secondary ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Genes, Reporter ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Lentivirus ; genetics ; metabolism ; Luciferases ; biosynthesis ; genetics ; Luminescent Measurements ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Transplantation ; Receptor, ErbB-2 ; metabolism ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism

The aim of this study was to evaluate the therapeutic effects and adverse reactions of fludarabine-based regimen for patients with chronic lymphocytic leukemia(CLL).18 patients with CLL were treated with F regimen [fludarabine 30 mg/(m(2)·d) intravenously for 3 d, repeatedly every 28 days]. 22 patients were treated with FC regimen [fludarabine 25 mg/(m(2)·d) plus cyclophosphamide 250 mg/(m(2)·d) intravenously for 3 d, repeatedly every 28 days]. The results showed that the rate of complete remission (CR), partial remission (PR) and overall remission (OR) reached 16.7%, 61.1% and 77.8% in the F regimen groups and 59.1%, 40.9% and 100% in the FC regimen groups (P < 0.05, P > 0.05 and P > 0.05), respectively. FC regimen resulted in significantly higher CR rate than that in single-agent fludarabine regimen. The main adverse reactions were myelosuppression and immunosuppression. No significant differences were found between the two regimens. FC regimen did not increase the rate of severe infections. It is concluded that FC regimen can give higher CR rate as compared with F regimen, fludarabine-based regimens is effective and safe first-line regimen for patients with CLL.
Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; administration & dosage ; Female ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell ; drug therapy ; Male ; Middle Aged ; Treatment Outcome ; Vidarabine ; administration & dosage ; analogs & derivatives

No abstract available.
Asian Continental Ancestry Group ; Humans ; Spastic Paraplegia, Hereditary