A fusion expression vector pPIC3.5K-PDGFR? was constructed to express recombinant receptor tyrosine kinase PDGFR? and the right Pichia pastoris transformants were screened on his-deficient plates and YPD-G418 plates by turns after electroporation of strain GS115, a high yield strain named M3 was screened. The strain M3 was cultured in a 5 L fermentor and His-GFP-PDGFR? fusion protein was purified by Ni2+ chelating affinity chromatography. One distinct peak was obtained after elution with 250 mmol/L imidazole. Fusion protein was proved to be 90.08 kD by western blotting, and have tyrosine kinase activity by ELISA. Results showed that the receptor tyrosine kinase PDGFR? was successfully expressed in P. pastoris and could be used as a target for small molecule selective inhibitors screening.

Purpose:To explore the preventive effect of hepatic arterial infusion (DDS) against recurrence of hepato- cellular carcinoma (HCC) after radical resection.Methods:From Jan,1996 to May,1998,287 patients of HCC after radical resection were randomly divided into four groups:intra-hepatic arterial infusion(97),intra-portal vein(80),intra-hepatic artery and portal vein (60),control (50).All patients received chemotherapy for two years and observed for postoperative recurrence of HCC.Results:The postoperative recurrence rate of HCC with intra-hepatic arterial infusion was significantly lower than that of intra-portal vein (P0.05).The 1～、2～ and 3～year survival rate of intra-hepatic arterial infusion was significantly higher than any of the other groups.Conclusions:Intra-hepatic arterial infusion (DDS) through gastro-duodenal artery can effectively pro- long the postoperative survival and decrease the post-operative recurrence rate of HCC.The preventive method of DDS through gastro-duodenal artery was safer and effective.

ObjectiveTo study possible mechanism through investigating the pathological and ultrastructural characters of secondary injury to perihematoma in intracerebral haemorrhage (ICH) rats.MethodsSprague Dawley male rats were subjected to ICH models. They were randomly divided into test group and control group. The rats in the test group were divided into 7 subgroups at 1h,3h,12h,24h,48h,72h and 7d after ICH; while those in control group were divided into 3 subgroups at 3h,24h,72h after saline injection. Each subgroup contained 5 rats. 2 rats from each group were stained by 2% triphenyltetrazolium chloride(TTC) to observe the pathological change.3 rats were picked up from each group to do optical microscope and electric microscope investigation on perihematoma tissue and ipsilateral cortex.ResultsHematoma tissue was demonstrated as black brown by TTC staining, no white infarcted area was detected around hematoma. In addition, there was a transitional zone between hematoma and normal tissue under microscopy; the involved tissue looked loose with varied edematous cells. Astrocytes appeared swollen and neural cells looked degenerated and necrosis. Meanwhile, capillary hyperplasia around hematoma with foot plate swollen were detected, no remarkable neural cells change was observed. 24 h after blood injection, astrocytes started to swell, part of them became degenerated and necrosis. Neural cells appeared mild degenerated and blood brain barrier were destroyed. 72 h after ICH, astrocytes showed highly swollen with neural cells degenerated.ConclusionSecondary injury to perihematoma has been identified and the pathological and ultrastructural changes have been observed.

OBJECTIVETo investigate whether the ERK/FoxO3a signal axis could induce the inhibitory effect of vitexin 1 (VB-1) in HepG2 cell proliferation.

METHODThe MTT method was adopted to observe the effect of different concentrations of VB-1 on human hepatoma carcinoma cell line HepG2 and immortalized human embryo liver cell line L-02. The cell growth was assessed by the clone formation assay. The protein phosphorylation levels of ERK1/2 and FoxO3a were measured by the western blot.

RESULTVB-1 inhibited the viability of HepG2 cell line in a concentration-dependent manner, with a weak effect on L-02 cell line. VB-1 could effectively inhibit the anchorage-dependent growth of HepG2 cells, and reduce the expression levels of pERK1/2 and pFoxO3a in a concentration-dependent manner. MEK1/2 inhibitor PD98059 could enhance VB-1' s effect in inhibiting HepG2 cell proliferation and ERK1/2, FoxO3a phosphorylation.

CONCLUSIONVB-1 inhibits the proliferative activity of hepatoma carcinoma cell line HepG2 by blocking the ERK/FoxO3a signal axis.

Apigenin ; pharmacology ; Carcinoma, Hepatocellular ; drug therapy ; genetics ; metabolism ; physiopathology ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; genetics ; metabolism ; Forkhead Box Protein O3 ; Forkhead Transcription Factors ; genetics ; metabolism ; Growth Inhibitors ; pharmacology ; Hep G2 Cells ; Humans ; Liver Neoplasms ; drug therapy ; genetics ; metabolism ; physiopathology ; Signal Transduction ; drug effects

Objective To investigate the adverse effects of taurine on rabbit corneal endothelial cells. Methods Six rabbits (12 eyes) were selected, and 6 histologic sections were prepared from each of the eyes. Rabbit corneal endothelial cells were cultured by explant culture method. Cells were innoculated on a 12-well tissue culture plate, 2%, 4%, 6%, 8% and 10% taurine solutions were added respectively (cells from the right and left eyes of the same rabbit were added the same concentration of taurine solution), and blank control was established. The growth of corneal endothelial cells was observed by inverted microscopy, and cell morphology on the 1st, 2nd, 4th, 6th and 8th day of culture was observed with Wright staining. Results Corneal endothelial cells cultured with 2%, 4% and 6% taurine solutions and those of blank control formed endothelial cell layers after culture for one week, and the cells exhibited hexagonal or round-like morphology. Corneal endothelial cells cultured with 8% taurine solution appeared to be undergrowth with small cell body on the 4th day, and cell death occurred on the 8th day. Corneal endothelial cells cultured with 10% taurine solution turned out to be undergrowth with small cell body on the 2nd day, and cell death had occurred. The same growth velocity and cell morphology were observed in the corneal endothelial cells from the right and left eyes of the same rabbit. Conclusion Taurine with concentration between 2% and 6% has no adverse effects on the growth of rabbit corneal endothelial cells.

The method was established for determination of geniposidic acid, chlorogenic acid, pinoresinoldiglucoside, which are three kinds of constituents of Eucommia ulmoides absorbed into the blood components. LC-MS/MS technique was applied to determine the blood components of the bloodstream after administration of E. ulmoides extract. At the same time, HPLC was used for detection of the ingredients content of the blood sample from 23 batches of E. ulmoides. The results showed that geniposidic acid, chlorogenic acid and pinoresinoldiglucoside are prototype into the blood in rats after oral administration of E. ulmoides extract, The linear range of geniposidic acid, chlorogenic acid and pinoresinoldiglucoside was good, and the average recoveries geniposidic acid, chlorogenic acid and pinoresinoldiglucoside were 98.69%, 100.8% and 98.39%, respectively. The method is simple and feasible with good reproducibility.
Animals ; Chlorogenic Acid ; blood ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Eucommiaceae ; chemistry ; Glycosides ; blood ; Iridoid Glucosides ; blood ; Lignans ; blood ; Male ; Plasma ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

Bridged-Ring Compounds ; poisoning ; Humans ; Male ; Middle Aged ; Poisoning ; therapy ; Treatment Outcome

We herein reported a 27-year-old woman with a right renal mass for two years. She underwent laparoscopic partial nephrectomy. Immunohistochemical examination of the specimen confirmed the diagnosis of solitary fibrous tumor by revealing its positive staining for cluster of differentiation (CD)34, epithelial membrane antigen (EMA), B-cell lymphoma-2 (Bcl-2) and CD99 in the tumor cells. No adjuvant treatment was carried out. The patient was in good health without local recurrence or metastasis during 2 years of follow-up. Laparoscopic partial nephrectomy for renal solitary fibrous tumor is an alternative treatment to radical nephrectomy. It can provide a good outcome. However, further follow-up and more cases of renal solitary fibrous tumor treated with laparoscopic partial nephrectomy are necessary to compare the oncological outcome with radical nephrectomy.

There are similarities between magnetotactic bacteria and Acidithiobacillus ferrooxidans (A. ferrooxidans) which isolated from Acid mine drainage(AMD). The weak magnetotaxis of some bioleaching bacteria isolated were found by microscope. A magnetophoresis apparatus was designed based on these weak magnetotaxis and be used to analysis the movement of these strains. The physiological properties of the anear magnetic field strain and removed magnetic field strain which isolated successfully by magnetophoresis apparatus have large difference. The nanometer magnetic particles was extract from the Acidithiobacillus ferrooxidans which purified by spread plate method from AMFS and its main elements are Fe and O by energy spectrum analysis. The results show that A. ferrooxidans have weak magnetotaxis and can be isolated by magnetophoresis. With the development of this new isolating method, the research of magnetotactic bacteria and bioleaching will get more benefit from it.

Objective To analyze the pathological and clinical characteristics of patients with idiopathic IgA nephropathy accompanied by vasculitic/crescentic lesion (IgA-V/C). Methods Data of 222 patients diagnosed as idiopathic IgA nephropathy by renal biopsy, among them 87 cases with vasculitic/crescenlic (V/C)lesion, from our department in 2004 were analyzed retrospectively. Clinical and pathological data from patients with IgA-V/C were compared to those of non-IgA-V/C patients and of lupus nephritis (LN) patients with V/C lesion. Results Vasculitic/crescentic lesion was found in 39.19% (87/222) patients with idiopathic IgA nephropathy.And about( 14.08?12.75)% of the glomeruli was affected. It should be taken into account that there was no significant differences of clinical manifestations including blood pressure, urinary protein excretion between IgA-V/C group and non-IgA-V/C group .except serum creatinine(Scr)level which was significantly higher in IgA-V/C group. In addition, only 37.9% of IgA-V/C patients presented high Scr level,thus the lesion of V/C in idiopathic IgA nephropathy was easily overlooked. Patients with idiopathic IgA nephropathy were found to have higher percentage of glomerular sclerosis (64.86% vs 40.00%) and ratio of sclerostic glomeruli to total glomeruli [ (26.98 ?24.68)% vs (16.10 ?18.80)% ]as compared to LN group, which further predicated the progressing characteristics of IgA nephropathy. Conclusions Vasculitic/crescentic lesion is a quite common finding in idiopathic IgA nephropathy and often associated with no dramatically symptoms. It is possible for vasculitic/crescentic lesion to induce unmarked lose of nephron slowly and continually, so as to accelerate IgA nephropathy progression to end-stage renal failure.