Purpose:To explore the effects and mechanisms o f anti-invasion and anti-metastasis of ginsenoside Rg3 on human hepatocellular carcinoma cells. Methods:To select human hepatocellular carcinoma cell line SMMC -7721 and the human allantoic veins endothelial cell line ECV304 as the study objects. We observed the effet of ginsenoside Rg3’s effect on the growth of SM MC-7721 and ECV304 by MTT methods,the adhesion of SMMC-7721 and Fibronectin by cell adhesion experiment, the expression of gene protein of nm23,CD44 and VEGF by immunohisto-chemical method. Results:Ginsenoside Rg3 could inhibit not only the growth of SM MC-7721 and ECV304 significantly, but also the adhesion of SMMC-7721 and FN. I t might also down-regulate the expression of CD44, VEGF and up-regulate the nm 23 gene expression. Conclusions:Ginsenoside Rg3 can inhibit invasion and metastasis of the hepatic carcinoma cell. The mechanisms are probably that Ginsenoside Rg3 can inhibit the hepatic carcinoma cells’ invasion activity,regulate the expres sion of the gene proteins which are closely related with invasion and metastasis ,inhibit neovasularization.

AIM: To analyze the bacterial resistance of clinical isolates in a hospital. METHODS: Susceptibility of 285 strains of G - bacillus and 133 strains of G + bacillus was observed in 18 kinds of antibiotics. RESULTS: Resistance increased in most G - bacilli to the third generation cephalosporin. The resistance rate was 28.4 % in the 74 strains of staphylococcus aureus, and 3.4 % in enterococci to vancomycin. No vancomycin resistant strain was found. Extend spectrum ? lactamases of E.coli and klebsiella (ESBLs) accounted for 34.0 % and 29.7 %, respectively. CONCLUSION: The drug resistance is severe in antibiotics, indicating that susceptibility determination is important in selection of antibiotics.

Objective To compare the features of magnetic resonance imaging (MRI) and electroencephalogram (EEG) to differentiate Alzheimer's disease (AD) and vascular dementia (VD). Methods It was analyzed of the MRI and EEG from 39 patients with AD and 56 patients with VD, to compare the proportion of cerebral atrophy, hippocampal atrophy and leukoaraiosis in MRI, and the proportion of the moderate to severe disorder of EEG and the power spectrum. Results The proportion of cerebral atrophy and hippocampal atrophy was more and leukoaraiosis was less in the AD group than those in the VD group. The proportion of the moderate to severe disorder of EEG increased in AD group, and the ratio of (θ+δ)/(α+β) of whole brain was more in the AD group than in the VD group (P<0.05). Conclusion It is more likely to be AD in dementia patients with atrophy without leukoaraiosis and cerebral ischemic lesions, especially for those with severe abnormal EEG.

In this paper, the biological compatibilities of FGF/Collagen compound sponges are evaluated. Intracutaneous stimulation test and shortterm muscle embedding test of FGF/Collagen compound sponges made of different materials are performed. Experimental data are analyzed and evaluated according to the standard. The two sponges get marks of 0 in acute eye stimulation test and intracutaneous stimulation test, thus they are not stimulating to tissues. In shortterm muscle embedding test, they don't lead to inflammation reactions and can be degraded and absorbed in short term. FGF/Collagen compound sponge proves good biological compatibility and has a cheerful prospect in medicine.

AIM: To investigate the pharmacological effects of recombined basic fibroblast growth factor (rbFGF) on retinal ganglion cells (RGCs) of rats. METHODS: Using calibrated cross-action forceps a moderate crush injury was inflicted on the nerve. After crush injury, rbFGF, saline and VB 12 were administered by retrobublar injection. Four weeks after injury , the apoptosis of RGCs was measured with flow cytometer. RESULTS: Four weeks after operation, it was shown that the rbFGF, but not saline or VB 12 injection could significantly improve the maintainance of RGCs of rats. After 800 U, 1600 U and 2400 U rbFGF injection, the injured RGCs were rescued by 24.5%, 27.3% and 28.5% respectively. Furthermore, it was also found that rbFGF injection could effectively prevent the axons from injury. The flow cytometer showed that the rate of apoptosis was reduced markedly on 7 days at rbFGF group. CONCLUSION: rbFGF can significantly promote the functional repair of injured optic nerve. [

AIM:To study the mechanism of the unique export of one of human basic fibroblast growth factor (hbFGF) forms lacking the N-terminal nuclear localization signal (NLS),we high expressed and purified this hbFGF form in E.coli strain BL21(DE3).METHODS:The cDNA fragment of the hbFGF amplified by polymerase chain reaction (PCR) was cloned into the expression vector pET3c and expressed in BL21(DE3) by IPTG induction.The expressed hbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate.The mitogenic activity was measured by MTT.RESULTS:The expression level of hbFGF in E.coli was about 20% of the total cellular protein.The appreciable mitogenic activity of the purified hbFGF was comparable to that of hbFGF standard.CONCLUSION:The BL21(DE3)/ pET3c expression system could be used to high express hbFGF lacking NLS.The purified recombinant hbFGF was prepared and sufficient for further study.

AIM:To synthesize the human platelet factor-4(hPF4) gene with a convenient and effective approach, and high express the hPF4 gene in E. coli BL21 (DE3). METHODS: According to the primary structure of hPF4, the nucleotide sequence was synthesized using touch-down PCR method. The resultant gene fragment containing EcoR Ⅰ and Xho Ⅰ overhangs at 5' and 3' ends was cloned into the expression vector pGEX-4T-1 to construct the recombinant plasmid pGEX-4T-1-hPF4,which was then transformed into the E. coli strain BL21 (DE3). RESULTS: hPF4 gene was successfully synthesized by touchdown PCR method. A fusion protein composed of glutathione S-transferase (GST) and the recombinant hPF4 was expressed in BL21(DE3) by IPTG induction. The expression level of the fusion protein in E. coli was about 30% of the total cellular protein. CONCLUSION: Touch-down PCR may provide a convenient and effective approach to obtain other target genes. The expressed fusion protein forms the inclusion bodies, providing sufficient material for further purification and biological activities process.

AIM: To observe the effects of the human acidic fibroblast growth factor mutant (mhaFGF), lacking 27 amino acids at N-terminal, on the proliferation and signal transduction of the hepatocarcinoma cells. METHODS: The hepatocarcinoma cells were treated with human acidic fibroblast growth factor (haFGF) and mhaFGF, respectively. The expression levels of the signal proteins, Grb2 and Erk1/2, in the hepatocarcinoma cells were detected by semi-quantitative Western-blotting after treated for 15 min. The mitogenic activity of both haFGF and mhaFGF was detected by MTT method and the cell cycle was analysed by flow cytometer (FCM) after treated for 48 h. RESULTS: The mitogenic activity and the ratio of G 1 and S phase cells in mhaFGF-treated cells were markedly lower than that of the haFGF, and close to that of the control group. The expression level of both Grb2 and Erk1/2 in the mhaFGF-treated cells were lower than those in the haFGF- treated cells. CONCLUSION: The decrease in the mitogenic activity of mhaFGF is probably associated to its down-regulating the expression of the signal molecular, MAPK-ERK1/2 and Grb2.

In order to decrease the potential side-effects of human basic fibroblast growth factor (hbFGF) caused by its broadspectrum mitogenic activity, a single residue of hbFGF, the residue serine 108, was replaced with neutral alanine residue to construct a mutant of hbFGF (mhbFGF) with reduced mitogenic activity. The mutant was overexpressed in Escherichia coli BL21(DE3) by IPTG induction. The expression level of mhbFGF was about 30% of the total cellular protein. The expressed mhbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate. Measured by MTT method, the effect of mhbFGF on Balb/c 3T3 cell proliferation was much lower than that of wild-type hbFGF. The purified recombinant mhbFGF was prepared and sufficient for the following pharmacological and safety studies.

OBJECTIVESTo investigate the expression of macrophage migration inhibitory factor (MIF) mRNA in Schwann cells after peripheral nerve injury and roles of Schwann cells and MIF in macrophages activation and nerve regeneration.

METHODSFifty SD rats were divided into 10 groups. One group served as normal control. The rest were anesthetized with 3% sodium pentobarbital (30 - 60 mg/kg, i.p) and sciatic nerves were transected distal to the obturator tendon respectively 1 h, 12 h, 1 d, 3 d, 7 d, 10 d, 14 d, 17 d and 21 d before being killed. Sciatic nerves were resected and connective tissues excised. Schwann cells were obtained by digesting the nerve tissues with trypsin and collagenase. RNA was isolated and reverse-transcription-polymerase chain reaction (RT-PCR) was carried out. cDNA was analyzed by automatic system and the parameters were assessed to define the status of MIF mRNA expression in different groups.

RESULTSThe level of MIF mRNA started to increase 12 h after the nerve transection. The level remained high from day 7 up to 10 after the injury. During the period from days 10 to 21, MIF mRNA decreased slowly to the pre-transection level.

CONCLUSIONAfter peripheral nerve injury, Schwann cells can secrete MIF which may play a pivotal role as an immunomodulatory cytokine in macrophage activation and inflammatory reaction.

Animals ; Female ; Macrophage Migration-Inhibitory Factors ; genetics ; Male ; Peripheral Nerve Injuries ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Schwann Cells ; metabolism