Objective To construct and express recombinant cecropin B-binding site of luteinizing hormone releasing hormone(CB-LHRH')gene,and to evaluate the anticancer function of CB-LHRH' on human ovarian cancer cell line SKOV3 and human endometrial cancer cell line HEC-1B.Methods The sequence of the cDNA encoding CB-LHRH' was designed,artificially synthesized,verified by DNA sequence analysis and expressed by Bac-to-Bac baculovirus expression system.The expression of CB-LHRH' proteins were identified by western dot blot using rabbit polyclonal antibody against LHRH as the primary antibody.To determine the anticancer effects of the CB-LHRH' protein,ovarian cancer cell line SKOV3 and endometrial adenocarcinoma cell line HEC-1B were treated by different doses of the CB-LHRH' protein.Cell growth inhibition assay was performed using the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)5[(phenylamino) carbonyl]-2H-tetrazolium hydroxide(XTT)kit at different times,and cell morphologic changes were observed under the inverted microscope.Results The inhibitory rate of proliferation by CB-LHRH' increased with the increase of dose and time respectively:SKOV3 cell,from(5.03?0.08)% to(53.24 ?1.22)%;HEC-1B cell,from(5.13?0.37)% to(56.16?1.08)%.The inhibitory effect on HEC-1B cell was stronger than that on SKOV3 cell(P

Objective To investigate the effect of chymase on the proliferation of rat cardiac fibroblasts (CFs) and the role of transforming growth factor-?1 (TGF-?_1).Methods Cultured CFs of neonatal SD rats were isolated by trypsinization.Cell number and DNA synthesis were evaluated by MTT assay (A_(490) value) and [~3H]-deoxythy- midine [~3H]-TdR incorporation.The mRNA expression of TGF-?_1 in CFs was determined by RT-PCR.Results Chymase increased CFs numbers and [~3H]-TdR incorporation in a dose-dependent manner.The A_(490) value of CFs stimulated by 15,30 and 60 ng/mL chymase was 0.263?0.033,0.348?0.031 and 0.387?0.026,respectively, which were all significantly higher than that of control (0.201?0.019,P

SUMMARY Objective:To investigate the influence of cryoprotectants and cooling rates in vitrificationmethod on the spindles of rabbit M Ⅱ oocytes.Methods:Rabbit oocytes were verified by using cryoloopwith ethylene glycol(EG)singly or EG combined with dimethyl sulphoxide(DMSO)as cryoprotectants,and cooled by taking oocytes directly into liquid nitrogen or by vitrification machine.After frozen rabbit o-ocytes thawed,the microtubulin and chromosome of the spindles were fixation and stained by immunofluo-rescent method.Confocal microscope was used to reveal spindle configuration.Results:In the two proto-cols of single EG used and EG combined with DMSO,the spindles were severely injured.But in protocolof EG combined with DMSO and at ultra-rapid cooling rate,the normal configuration of spindle rate ofthawed rabbit oobytes was similar to that of the control group.Conclusion:The protocol of EG combinedwith DMSO as cryoprotectants and with extremely high cooling rate by vitrification machine can producethe best effect on conservation of spindle configuration in vitrification of rabbit oocytes.

OBJECTIVETo study 3 different strategies of urine drainage following hypospadias urethroplasty, the clinical nursing in their application, and their effects.

METHODSWe retrospectively analyzed the clinical data of 595 cases of hypospadias treated by urethroplasty. After surgery, 133 of the patients underwent urine drainage by suprapubic cystostomy (group A), 202 by urethral stent- tube indwelling (group B), and 260 by early initiative micturition with the urethral stent-tube (group C). All the patients received routine postoperative nursing care required for hypospadias repair.

RESULTSOperations were successfully completed in all the cases. Group C showed a remarkably shorter hospital stay and lower incidence rates of urinary fistula and urethral stricture than groups A and B (P<0.05), but there were no significant differences in the three indexes between A and B (P<0.05).

CONCLUSIONFor urine drainage following hypospadias repair, early initiative micturition with the urethral stent-tube can significantly reduce postoperative complications, decrease difficulties and workload of nursing care, and shorten the hospital stay of the patient.

Cystostomy ; Drainage ; methods ; Humans ; Hypospadias ; surgery ; Length of Stay ; Male ; Postoperative Complications ; prevention & control ; Reconstructive Surgical Procedures ; Retrospective Studies ; Stents ; Urethra ; surgery ; Urethral Stricture ; prevention & control ; Urinary Fistula ; prevention & control ; Urine ; Urologic Surgical Procedures, Male

The study was aimed to investigate the effects of emodin on the proliferation and apoptosis of adriamycin-resistant HL-60/ADR cells, and to explore the underlying mechanism. The cell viability and colony formation were detected by MTT assay and colony formation assay respectively. Apoptotic cells were tested by means of cell cycle analysis, mitochondrial transmembrane potential levels, caspase-3 activity detection, Annexin V FITC/PI staining and TUNEL labeling. RT-PCR was used to analyze the bcl-2 and c-myc mRNA expressions. The protein expressions of Bcl-2, c-Myc and caspase-3 precursor were determined by Western blot. The results showed that HL-60/ADR cell growth was significantly inhibited by emodin in dose and time dependent manners. Cell colony formation obviously decreased with IC50 5.79 micromol/L. G0/G1 phase cell population increased while G2/M phase cells decreased in 40 and 80 micromol/L groups compared with control group (p < 0.01), and no significant difference of cell cycle was observed in 20 micromol/L group (p > 0.05). The typical hypo-diploid peak (apoptotic peak) appeared in each dose group. The levels of mitochondrial transmembrane potential of HL-60/ADR cells decreased and caspase-3 activity increased when incubated with emodin for 12 and 24 hours respectively. Apoptosis occurred in a dose-dependent manner, and its earlier and later stages were identified by Annexin-V FITC/PI staining and TUNEL labeling methods respectively. The expressions of bcl-2, c-myc mRNA and Bcl-2, c-Myc, caspase-3 precursor protein were all down-regulated in a time-dependent manner after treatment with emodin at different times. It is concluded that emodin efficiently inhibits growth and induces apoptosis on HL-60/ADR cells, which may be related with the down-regulation of mitochondrial transmembrane potential and expressions of bcl-2 and c-myc, as well as up-regulation of caspase-3 activity.
Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; drug effects ; Emodin ; pharmacology ; HL-60 Cells ; Humans

Pseudomonas sp. ZD8 isolated from contaminated soil was immobilized with platane wood chips to produce packing materials for a novel biofilter system utilized to control restaurant emissions. The effects of operational parameters including retention time, temperature, and inlet gas concentration on the removal efficiency and elimination capacity were evaluated. Criteria necessary for a scale-up design of the biofilter was established. High and satisfactory level of rapeseed oil smoke removal efficiency was maintained during operation and the optimal retention time was found to be 18 s corresponding to smoke removal efficiency greater than 97%. The optimal inlet rapeseed oil smoke loading was 120 mg/(m(3) x h) at the upper end of the linear correlation between inlet loading and elimination capacity.
Air Pollutants ; isolation & purification ; Biodegradation, Environmental ; Cells, Immobilized ; Filtration ; instrumentation ; methods ; Microscopy, Electron, Scanning ; Oils ; metabolism ; Pseudomonas ; metabolism ; ultrastructure ; Restaurants ; Smoke ; Temperature ; Time Factors ; Waste Management ; instrumentation ; methods

OBJECTIVETo probe the expression level of insulin-like growth factor-I receptor (IGF-IR) in malignant clone cells of myelodysplastic syndromes (MDS).

METHODThe combination of fluorescence in situ hybridization (FISH) and immunochemistry (alkaline phosphatase anti-alkaline phosphatase, APAAP) was used to detect the expression of IGF-IR in the bone marrow cells of 40 MDS patients with abnormal karyotypes.

RESULTSThe average IGF-IR expression level on the clone cells \[(84.5 ± 14.2)%\] from the MDS cases was markedly elevated compared to the corresponding level on normal cells \[(11.4 ± 12.1)%\]. And the percentages of malignant clone cells in all 40 MDS cases were significantly correlated with the relevant percentages of IGF-IR positive nucleated cells (r = 0.929, P < 0.01).

CONCLUSIONSIGF-IR might be taken as a marker of clone cells in MDS for its trait to malignant proliferation.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Clone Cells ; metabolism ; pathology ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; metabolism ; pathology ; Receptor, IGF Type 1 ; metabolism ; Young Adult

OBJECTIVEThe study was aimed to investigate the inhibitory effect and mechanism of astragaloside IV (ASI) on the activation of microglial cells.

METHODAfter pre-incubated with ASI for 2 h, microglial cells BV-2 were stimulated with interferon-γ (IFN-γ) for 1. 5 h and 24 h, respectively. Secretion of nitric oxide (NO) in the medium was measured by Griess method. Production of tumor necrosis factor alpha (TNF-α) was detected by ELISA approach. Cellular gene expressions of CD11b, TNF-α, interleukin 1β (IL-1β) and induced nitric oxide synthase (iNOS) were examined by quantitative-PCR analysis. Total and phosphorylation of STAT1, IκB and NF-κB was analyzed by Western blot method.

RESULTASI could significantly inhibit the increased secretion of TNF-α and NO from BV-2 cells upon IFN-γ stimulation (P < 0.001). Further study showed that ASI significantly down-regulated gene expression of IL-1β and TNF-α (P < 0.01, P < 0.05) and exhibited a trend to reduce that of iNOS. IFN-γ and ASI have no obvious effect on gene expression of CD11b. Moreover, ASI inhibited the phosphorylation of STAT1, IκB and NF-κB elicited by IFN-γ stimulation.

CONCLUSIONASI could restrain microglial activation through interfering STAT1/IκB/NF-κB signaling pathway, reducing gene expres- sion of IL-1β and TNF-α, and thus inhibiting the production of proinflammatory mediators such as NO and TNF-α.

Animals ; Astragalus Plant ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; I-kappa B Proteins ; genetics ; metabolism ; Interferon-gamma ; genetics ; metabolism ; Mice ; NF-kappa B ; genetics ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; STAT1 Transcription Factor ; genetics ; metabolism ; Saponins ; pharmacology ; Signal Transduction ; drug effects ; Triterpenes ; pharmacology

Objective To investigate the mechanism and suppression effect of human cytomegalovirus (HCMV) on proliferation of megakaryocyte progenitors(CFU-Mk)in vitro.Methods Colony forming unit-assay was applied to observe the effect of HCMV-AD 169 strain on CFU-Mk of cord blood. The technique of PCR was used to demonstrate the existence of HCMV-AD 169 DNA in the colony cells of cultured CFU-Mk.Results 1.The number of CFU-Mk colonies in HCMV-infected groups decreased significantly compared with that of control group. The CFU-Mk formation was inhibited significantly after HCMV-AD 169 strain infection.The suppression effect showed a dose-dependent fashion: 46.7 % inhibition with 10 -1of HCMV, 29.7 % with 10 -2 and 14.5 % with 10 -3 in the CFU-Mix assay. The peak of CFU-Mk colonies (d16-18) was not significantly different between control group and experimental groups, but the duration of the CFU-Mk colonies in infected groups was significantly shorter than that in control group.2. HCMV-DNA was positively detected in the colony cells of viral infected group by PCR, while negative in control group.Conclusions HCMV-AD 169 strain may inhibit the differentiation and proliferation of CFU-Mk by infecting the hematopoietic progenitors. HCMV may cause the suppression of hematopoiesis by direct infection, which may be the main reason for HCMV infection associated with thrombocytopenia.

As a therapeutic agent in thrombosis the fibrinolytic enzymes are of interest and the search for a new enzyme continues. A novel fibrinolytic enzyme was produced from Rhizopus chinensis 120, which was screened from the starter for brewing rice wine in the South of China, by solid fermentation, and purified through ammonium sulfate precipitation, hydrophobic interaction, ionic exchange and gel filtration chromatographies. The purified enzyme hydrolyzed fibrin, it cleaved the alpha-, beta- and gamma-chains of fibrinogen simultaneously, and it also activated plasminogen to plasmin. The enzyme hydrolyzed N-Succinyl-Ala-Ala- Pro-Phe-pNA, and Km was 0.23 mmol/L and Kcat 16.36 s(-1). The optimal temperature of the enzyme for hydrolying fibrin was 45 degrees C, and the optimal pH range of 6.8 - 8.8. The isoelectric point of the enzyme estimated by isoelectric focusing electrophoresis was 8.5 +/- 0.1. The enzyme was a glycoprotein. EDTA, PCMB, PMSF inhibited the activety of the enzyme, and SBTI, Lys, TPCK, Aprotinine had none obvious inhibition, which suggested that the activity centre of the enzyme had hydrosulfuryl, metal and serine. The first 12 amino acids of the N-termimal sequence of the enzyme were NH2-Ser-Val-Ser-Glu-Ile-Gln-Leu-Met-His-Asn-Leu-Gly, and had none homology with that of other fibrinolytic enzyme from other microbes. The novel fibrinolytic enzyme from Rhizopus chinensis 12# has potential to become a therapeutic agent in thrombosis.
Enzyme Stability ; Fermentation ; Fibrinolysin ; metabolism ; Fibrinolysis ; Fibrinolytic Agents ; chemistry ; Humans ; Plasminogen ; metabolism ; Rhizopus ; enzymology