BACKGROUND: It has been proved that the application of basic fibroblast growth factor (bFGF) combined with Brucea Javanica oil emulsion can accelerate wound healing and inhibit scar formation.OBJECTIVE: To observe the effects of bFGF plus Brucea Javanica oil emulsion cream on accelerating the skin wound healing of rabbits.DESIGN: A randomized controlled observation.SETTING: Center of Biotechnological Research and Development, Jinan University; College of Pharmacy, Jinan University.MATERIALS:The experiment was carried out in the College of Pharmacy, Jinan University from June to September in 2004. Eight Beijing big-ear white rabbits (4 males and 4 females) of 2.0-2.5 kg were provided by the experimental animal center of Southern Medical University (certification number: SoKx-2002-010). bFGF sterile freeze dried powder agent,provided by Guangzhou Changsheng Gene Pharmaceutical Co.,Ltd (batch number: 20040219; specific activity was 6 000 U/bottle), was prepared to solution with water for injection before application. Brucea Javanica oil emulsion (manufactured by Zhejiang 999 Bang'erkang Pharmaceutical Co.,Ltd.) was provided by Professor Yao from staff Room of Pharmacy, Shenyang Pharmaceutical University.METHODS: The rabbits were anesthetized and disinfected, 5 round wounds with diameter of 1.8 cm and area of 2.54 cm2were induced from front to back by bilateral incision at 1.5 cm from middle spine of rabbit. The 5 wounds of each rabbit were randomly divided into bFGF-treated group (90 U/cm2), bFGF+Brucea Javanica oil emulsion group [the wound was smeared with Brucea Javanica oil emulsion (30 mg/cm2) 30 minutes after bFGF (90 U/cm2)], Brucea Javanica oil emulsion treated group [the wound was smeared with Brucea Javanica oil emulsion (30 mg/cm2)], blank emulsion group (30 mg/cm2) and blank control group (the wound was smeared with saline). The medication was give immediately after injury, and changed once a day for 16 days. At 4, 8, 12 and 16 days after injury, the wound areas were recorded with the method of hyaline membrane tracing (the wound was covered with clean saran wrap, the size of wound was traced,and then sheared to be weighed, and converted to calculate the area), and the volume of wounded cavity was measured by infusing water. At 8 and 16 days, the wound tissue was removed, stained after routine tissue sections, and the conditions of growth of granulation tissue and reepithelization on the wound surface were observed.MAIN OUTCOME MEASURE: The wound area, volume of wounded cavity, and the conditions of growth of granulation tissue and reepithelization on the wound surface were obviously at different time points after injury in each group.RESULTS:All the 8 rabbits were involved in the analysis of results. ① Wound areas at different time points in each group: The wound areas in the bFGF+Brucea Javanica oil emulsion group at 4, 8 and 12 days after medication were smaller than those in the blank control group at corresponding time points [(2.05±0.35), (1.59±0.25), (0.55±0.25) cm2;(2.53±0.30), (2.41±0.19), (1.09±0.34) cm2, P＜0.05-0.01]. The wound areas in the bFGF group at 8 and 12 days after medication were (1.71±0.31) and (0.51±0.10) cm2, which were significantly smaller than those in the blank control group at corresponding time points (P＜0.05-0.01). ② Volume of the wounded cavity in each group: The wound volume in the bFGF group at 4 days after injury was markedly smaller than that in the blank control group at corresponding time point [(0.49±0.12), (0.59±0.1) mL, P＜0.05]. The wound volumes in the bFGF+Brucea Javanica oil emulsion group at 4 and 8days after injury were significantly smaller than those in the blank control group at corresponding time points [(0.47±0.12), (0.30±0.08) mL; (0.59±0.1), (0.41±0.07) mL, P＜0.05, 0.01]. ③ Growth of granulation tissue and reepithelization on the wound surface in each group: At 8 days after injury, the inflammatory reaction was milder and fibroblasts proliferated significantly in the bFGF+Brucea Javanica oil emulsion group, and the numbers of capillary plumules and fibroblasts were significantly more than those in the blank control group. The conditions in the blank control group and blank emulsion group were generally the same that there were severe inflammatory reactions, obvious increase of granulation tissue, fewer new capillaries, and unobvious proliferation of epidermic cells. At 16 days after injury, the contraction and reepithelization on the wound surface were obvious, and the new epithelia went towards the wound center rapidly in the bFGF+Brucea Javanica oil emulsion group.CONCLUSION : The application of Brucea Javanica oil emulsion plus bFGF can obviously accelerate the repair of incised wound on the back of rabbits.

Objective To evaluate the clinical outcome and relative factors of intervention treatment for atherosclerotic renal artery stenosis in elderly patients.Methods The clinical data of 79 patients diagnosed as atherosclerotic renal artery stenosis by angiography and treated by revascularization were analyzed.Results There were 55(69.6%)successes and 24(30.4%)failures in decreasing blood pressure and 28(35.4%)successes and 51(64.6%)failures in improving renal function after intervention treatment.Predictors of favorable outcome of intervention treatment in decreasing blood pressure were related to lower urine protein,higher glomerular filtration rate,higher systolic and diastolic blood pressure before treatment,lower resistance index(RI)of renal artery,and no complication of cerebral vascular diseases.Predictors of favorable outcome of intervention treatment in improving renal function were related with percentage of angiographic stenosis,category of antihypertension and lower urine protein.The logistic regression analysis showed that the percentage of angiographic stenosis was the most important predictor of intervention treatment for blood pressure control,age and level of serum creatinine before intervention treatment were the most important predictors of intervention treatment for improving renal faction.Conclusion Percentage of stenosis(≥85%),age(133 ?mol/L)can be used as the predictors of therapeutic success for renovascular stenosis in older patients.

As a new type of pesticides and because of their high performance and low toxicity, pyrethroid insecticides are widely used in place of organochlorine insecticides both in agriculture and in the home. In the recent years, more and more evidence indicates that pyrethroid insecticides can reduce sperm count and motility, cause deformity of the sperm head, increase the count of abnormal sperm, damage sperm DNA and induce its aneuploidy rate, as well as affect sex hormone levels and produce reproductive toxicity. The present article reviews the advances in the studies of male reproductive toxicity of pyrethroid pesticides by experiment in animals and human population, discusses the mechanism of male reproductive toxicity of pesticides and raises some problems concerning the evaluation of human reproductive hazards.
Animals ; Genitalia, Male ; drug effects ; pathology ; physiopathology ; Humans ; Insecticides ; poisoning ; toxicity ; Male ; Mice ; Pyrethrins ; poisoning ; toxicity ; Rats ; Toxicity Tests

OBJECTIVETo study the anticancer effect in vitro and in vivo and mechanism of DHA compound.

METHODSCervical cancer cell line HeLa cells, glioma cell line U251 cells and mouse hepatoma H(22) tumor were used in this study. Transmission electron microscopy and fluorescence microscopy were used to observe the morphological changes of cell apoptosis. Western blot was used to detect the expression of caspase-3. RT-PCR was used to determine the effect on Bcl-2 and Bax mRNA transcription in U251.

RESULTSAntitumor effect was observed in vivo and in vitro. Typical morphological changes were seen in cancer cells. The level of caspase-3 was significantly increased and the content of Bcl-2 mRNA was decreased significantly, while the content of Bax mRNA was significantly increased in the U251 cells after treatment with DHA compound.

CONCLUSIONDHA compound can inhibit the growth of some types of tumors and the increase of caspase-3 and Bax mRNA and decrease of Bcl-2 mRNA may be involved in its mechanism of action.

Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Docosahexaenoic Acids ; pharmacology ; Glioma ; pathology ; HeLa Cells ; Humans ; Liver Neoplasms, Experimental ; metabolism ; pathology ; Male ; Mice ; Neoplasm Transplantation ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

Based on site-specific transposition of an expression cassette into a baculovirus shuttle vector (Bacmid) which propagated in Escherichia coli, the Bac-to-Bac System provides a rapid and efficient method to generate recombinant baculoviruses and is widely used for high level expression of heterologous proteins. And the efficiency of recombinant baculovirus infecting cells plays an important role on the protein expression. In this study, we introduced an EGFP expression cassette driven by polyhedrin promoter into the p74 locus of Bacmid by homologous recombination. The target Bacmid-egfp was then transformed into E. coli DH10B containing the transposition helper plasmid to gain a new transposition receipt strain E. coli DH10Bac-egfp. Because of the intact attTn7 sites and lacZ', target gene cloned in a pFastBac vector can be transposed into the Bacmid-egfp shutter vector to construct recombinant baculovirus, which would allow the tracing of the target protein expression and the recombinant Bacmid transfection or recombinant baculoviral infection under fluorescence microscopes. Recombinant virus Bac-egfp-DsRed was constructed by transposing DsRed into the Bacmid-egfp in E. coliDHl0Bac-egfp, and the Sf9 cells infected with the recombinant virus expressed DsRed and EGFP efficiently. Another protein IL-6 fused with 6 x his tag was expressed and purified sucessfully from Sf9 cells infected with recombinant virus Bac-egfp-6 x his-IL6 constructed by the improved Bac-to-Bac system.
Animals ; Baculoviridae ; genetics ; Green Fluorescent Proteins ; genetics ; Interleukin-6 ; biosynthesis ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Spodoptera

OBJECTIVETo investigate hereditary susceptibility to coronary heart disease (CHD) in apolipoprotein E(apo E) and apo B polymorphisms of youths.

METHODSPolymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to analyze apoE, apoB Xba I, apoB 3' variable number of tandem repeat (VNTR) genotypes for 244 healthy Han students (among them were 109 students with positive CHD family history).

RESULTSThe allele frequencies of apo e4, XbaI x(+), 3'VNTR-B(hypervariable element, HVE>38) in the positive group were obviously higher than those in the negative group(P<0.05), and were significantly correlated with the increase in TC, LDL-C, apoB100 levels (P<0.05).

CONCLUSIONThe alleles for apo e4, XbaI x(+), 3'VNTR-B may be the important genetic markers of Han CHD.

Adolescent ; Alleles ; Apolipoproteins B ; genetics ; Apolipoproteins E ; genetics ; Coronary Disease ; genetics ; Female ; Gene Frequency ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Young Adult

To explore anti-tumor active components of Chimonanthus salicifolius, the phytochemistry of the chloroform fraction from leaves extract was investigated by repeated silica gel column chromatography. Twelve compounds were isolated and their structures were identified by physicochemical properties and spectroscopic data analysis as 9-epi-blumenol C(1), blumenol C(2), (+)-dehydrovomifoliol (3), (+)-vomifoliol (4), robinlin (5), (-)-loliolide (6), isofraxidin (7), scopoletin (8), 6,7-dimethoxycoumarin (9), 6, 7, 8-trimethoxycoumarin (10), beta-sitostenone (11), and beta-stigmasterol(12). Compounds 1-6 belonging to nor-sesquiterpenoids were isolated from the family Calycanthaceae for the first time. Compound 1 was a new natural product. Compounds 7, 11 and 12 were obtained from this plant for the first time.
Antineoplastic Agents ; analysis ; isolation & purification ; Calycanthaceae ; chemistry ; Chloroform ; chemistry ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Plant Leaves ; chemistry

OBJECTIVETo compare the effects of different doses of microwave on the proliferative activity and cell cycle of cultured epithelial cells of rabbit lens, and to investigate the limit tolerant of microwave exposure.

METHODSCultured epithelial cells of rabbit lens were exposed to microwave radiation with frequency of 2,450 MHz and power density of 0.10, 0.25, 0.50, 1.00, 2.00 mW/cm(2) for 8 h in vitro. HE staining was used to observe the morphological changes of lens epithelial cells, the proliferative activity and cell cycle were measured by MTT assay and PI fluorescent staining.

RESULTS8 h after radiation, 0.50, 1.00 and 2.00 mW/cm(2) microwave could decrease the proliferation of lens epithelial cells, make the cells disordered arrangement, shrinkage, detachment, and inhibit the synthesis of cell DNA. The percentage of G(0)/G(1) phase cells were 71.95% +/- 2.12%, 75.68% +/- 3.35% and 82.40% +/- 8.68% respectively, which were higher than that in control group (61.68% +/- 5.76%, P < 0.05 or P < 0.01). The percentage of S phase cells were 19.32% +/- 3.07%, 16.08% +/- 4.91% and 12.98% +/- 8.08% respectively, which were lower than that in control group (28.05% +/- 5.12%, P < 0.05 or P < 0.01). No obvious changes could be detected in 0.10, 0.25 mW/cm(2) microwave groups (P > 0.05).

CONCLUSIONMicrowave exceeding 0.50 mW/cm(2) may make injury to lens epithelial cells after 8 hour radiation, which may be related to the effect of microwave radiation on cell cycle.

Animals ; Cell Cycle ; radiation effects ; Cells, Cultured ; DNA ; metabolism ; Epithelial Cells ; cytology ; metabolism ; radiation effects ; Lens, Crystalline ; cytology ; metabolism ; radiation effects ; Microwaves ; Rabbits

Recombinant adeno-associated viruses (rAAV) vectors have been shown to mediate long-term transgene expression in mice and nonhuman primates. We have adapted viral vector system based on two rAAV vectors, namely rAAV1 and rAAV2. We have generated rAAV vectors expressing the envelope glycoprotein (E1 and E2) derived from Chinese HCV patient (genotype 1b) and used these to immunize BALB/c mice. We detected the total antibody titer by IFA and neutralizing antibody (nAb) using in vitro HCV neutralizing assays based on HCV pseudotyped particles. Furthermore, IFN-gamma ELISpot assay was used to assess the T cellular response against HCV at 12 weeks after rAAV1-E1E2 immunization. We also analyzed HCV envelope glycoprotein expression in muscle of rAAV1-E1E2 immunized mice. Our data showed: (i) rAAV1 directed long-term expression of HCV genes in mice; (ii) immunized intramuscularly with a single dose of rAAV elicited durable and effective immune responses in mice; and (iii) Moreover, rAAV1-E1E2 induced higher total antibody and nAb titers than rAAV2-E1E2 did. These data suggest that rAAV1 vectors could stimulate robust, durable, and effective immune responses against HCV.
Animals ; Antibodies, Viral ; blood ; Dependovirus ; genetics ; metabolism ; Female ; Genetic Vectors ; genetics ; metabolism ; Hepacivirus ; genetics ; immunology ; Hepatitis C ; immunology ; virology ; Humans ; Mice ; Mice, Inbred BALB C ; Vaccines, DNA ; administration & dosage ; genetics ; immunology ; Viral Envelope Proteins ; administration & dosage ; genetics ; immunology ; Viral Vaccines ; administration & dosage ; genetics ; immunology

In this study, three expression vectors encoding unmodified glycoproteins E1 and E2 from H77 (1a), Hebei (1b) and JFH1 (2a) strains were constructed to form pVRC-H77-E1E2, pVRC-HeBei-E1E2 and pVRC-JFH1-E1E2 expressing constructs. The protein expression was confirmed by immunofluorescene assay(IFA) and Western blot. The Lentiviral vector has the ability to package the cellular membrane into pseudo-particles. The plasmid expressing HCV E1-E2 glycoproteins in native form was co-transfected into 293FT cells with a lentiviral packaging plasmid (pHR'CMV delta R8.2)and a self-inactivated (SIN) transfer plasmid (pCS-CG) containing a reporter EGFP gene to produce infectious HCV pseudo-particles(pp). Flow cytometry assays showed that the HCVpp could infect Huh7 and Huh7-CD81, and the infectivity in Huh7-CD81 was about 2-3 times higher than that in Huh7 cells. Meanwhile, HCVpp could neither infect non-liver cells, for example, the 293 cells, nor HepG2 cell . Titration of HCVpp by p24 ELISA assay or infection assay showed that this HCVpp may contain 5-25 ng/mL p24 or 10(4)-10(5) TU (transducing unit)/ ml. An in vitro HCV neutralizing assays based on HCVpp (1a, 1b, 2a) were then established using AP33, a monoclone antibody with cross-neutralizing ability to different HCV strains. The neutralizing ability of the antibodies from HCV infected patients was further studied with this HCVpp system. In summary, three kinds of HCVpp (1a, 1b, 2a subtype) were successfully developed; In vitro HCV neutralizing assays based on HCVpp and SIN lentiviral system were established. This system paves a way for characterization of early steps of HCV infection (host tropisms, receptor binding, membrane fusion, et al. ) or screening anti-HCV drugs (such as inhibitor to virus entry). This system can be further applied to assess the human immune responses in HCV patients or evaluate HCV vaccine candidates.
Hepacivirus ; immunology ; Hepatitis C Antibodies ; immunology ; Hepatitis C, Chronic ; immunology ; Humans ; Neutralization Tests ; Viral Envelope Proteins ; immunology ; Virion ; immunology