Objective To explore the effect of fibroblast growth factor receptor(FGFR)inhibitor BGJ398 on the vasculogenic mimicry(VM)formation of glioma cells. Methods The phosphor-FGFR(pFGFR)was de-tected by Western blot,the expressions of MMP2 and MMP14 were detected by Western blot and immunocytochem-istry;the VM formation of U87MG and U251MG was tested by tube formation assay;subcutaneously implanted tu-mor model in nude mouse was established and tumor sections were CD34/PAS double-stained to detect the forma-tion of VM in vivo. Results Western blot showed that pFGFR in the experimental groups decreased significantly (P < 0.05);western blot and immunohistochemical staining showed that the expression of MMP2 and MMP14 in the experimental groups decreased significantly compared to the control group. In the tube formation assay ,the tube formation of U87MG and U251MG cells were restrained. In the subcutaneously implanted tumor model ,the VM number of the experimental group(13.85 ± 3.96)was significantly lower than that in the control group(26.40 ± 5.06,P < 0.05). Conclusions In vivo and in vitro experiments confirmed that BGJ398 can inhibit the activa-tion of FGFR,and inhibit the VM formation of glioma cells. These indicate FGFR signaling pathway is involved in the formation of VM.

Objective To study the impact of QseBC on the motility of Salmonella enterica serovar Typhi ( S.Typhi ) . Methods The motility of wild-type ( WT) and null mutants (ΔqseB and ΔqseC) at mid-log phase was investigated by swimming assay.Quantitative RT-PCR was carried out to calculate the transcriptional variation of flhD and qseB among WT,ΔqseB andΔqseC.QseB overexpressing strain was constructed to compare its motility and flhD expression with the wild-type control.Results The result of motility assay showed that the motility of ΔqseB was similar to that of the WT strain , while the motility of ΔqseC was much lower than that of WT .qRT-PCR revealed that compared with WT , the expression of flhD was significantly decreased in ΔqseC while the expression of qseB was increased considerably .The motility of QseB overex-pressing strain was lower .Conclusion The expression of flhD may be regulated by QseBC which has an effect on the motil-ity of S.typhi, and the overexpression of QseB may inhibit the motility .

Twenty-fine samples of stomach and 25 samples of adenocarcinoma stomach were employed for this study. The former have been collected from the patients of duodenal ulcer. Specimens were taken from the large curvature and pylorus. In the adenocarcinoma of stomach, materials were taken from the cancer focus and the cancer margins, fixed in 10% formalin-GaCl_2 Solution, embedded in paraffin and cut at 5? in thickness. Then they were examined with defferent reactions of histochemistry; PAS, Alcian blue (AB), Toludine blue (TB) and Barnett's method. The surface of gastric mucosa obtained from patients of duodenal ulcer after subtotalgastrectomy was covered with a substance which consists of neutral and acid mucopolysaccharide and protein. The epithelial cells of the mucosa contains large amount of neutral muco-polysaccharide. The cells of the neck of gastric gland contain quite a bit of acid muco-polysaccharide, but the cells of the deep segments of the glands do not show any of it. The cells of adenocarcinoma of stomach contain a different deal material mixture which consists of acid muco-polusaccharide and protein. The surface of the cavity in the well-differenlialed cancer glands was covered by a positive substance in TB, AB method, which forms a thin layer of membrance. An evenly distributed substance in the cavity of carcinoma gland was shown with positive reactions of PAS, AB, TB and protein. The degenerated cells show a positive reactions of PAS and protein. The cells of low-differentiated cancer were surrounded with a superficial layer of the acid mucopolysaccharide. The interstitia in the invaded margines often showed clear, distinct positive PAS and TB reactions. It indicates that it cortains more chondroitin sulfuric acid and the hyaline acid. In the superficial layers of mucosa of the cancer tissue, the epithelial cellswere often to have less positive substance than that of the membrane in the parts of cancer margine, it means a lesser amount of neutral-polysaccharide. But the acid muco-polysaccharide in the surface of cancer and its metaplastic margin clearly increases in amount than the metaplastic area far away from the cancer focus. This report discussed the significance of the increased muco-polysaccharide in the tissue of the gastric cancer.

Objective:To investigate the regulation mechanisms of the bone marrow mesenchymal stem cells on lymphocyte pro -liferation of type I diabetic rats .Methods:The rat bone marrow mesenchymal stem cells were isolated , cultured and identified and the effect on lymphocyte proliferation of type Ⅰdiabetic rat was observed by MTT assay , and analyze the CD 4 +CD25 +regulatory T cell ra-tio, cell cycle and apoptosis of type I diabetes rat by flow cytometric .Results:B and C groups was significantly lower than the absor-bance values of group A,the differences between the data were statistically significant (P<0.05), C group was significantly lower than group B absorbance values, the difference was significant (P<0.05);the CD4 +CD25 +regulatory T cells of B and C groups were sig-nificantly higher than group A, the differences of the data were statistically significant (P<0.05), the CD4 +CD25 +regulatory T cell ratio of C group significantly higher than that group B , the differences were statistically significant (P<0.05);the apoptosis levels of B and C groups were significantly higher than group A , the differences were statistically significant (P<0.05), the apoptosis levels of C group were significantly higher in group B , the differences were statistically significant (P<0.05).Conclusion:Bone marrow mesen-chymal stem cells can significantly inhibit lymphocyte proliferation of type Ⅰdiabetic rats, and it may regulate CD4 +CD25 +regulatory T cells, promote apoptosis, thereby affecting the immune function of T lymphocytes , and play its rejection.

Objective To optimize the preparation of nanoparticles encapsulating antisense oligodeoxynucleotides in a-butyleyanoacrylate carrier (ASODN in NP) and investigate their stability. Methods ASODN in NP were prepared by interfacial polymerization of butyleyanoacrylate (BCA). The formulation and technology of the prepared NP was optimized by using orthogonal design based on the single-factor experiment. The morphology of NP was examined by transmission electron microscope; The size and size distribution of NP were determined by Malvern laser granularity equipment;The encapsulation efficiency and drug loading were determined by HPLC; The ability of protecting oligodeoxynucleotides from serum was investigated on a 20% polyacrylamide-7 Murea sequencing gel (PAGE). Results The nanoparticles in the optimal conditions were of regular spherical surface and discrete. The average size was 97.1 nm,the average encapsulation efficiency and drug loading of ASODN in NP were 96.7% and 10.1% respectively; The oligonucleotides were more efficiently protected from degradation by nucleases than by oligonucleotides adsorbed into nanospheres.Conclusion ASODN in NP has good stability,encapsulation efficiency,drug loading and great potential for ASODN delivery.

OBJECTIVE To investigate the produce of extended-spectrum ?-lactamases(ESBLs) and the presence of genotype of the ?-lactamases-encoding genes in Klebsiella pneumoniae isolated from the 98th Hospital of PLA,Huzhou,Zhejiang Province,China.METHODS Twenty-five strains of K.pneumoniae were isolated from the inpatients between Sep 2005 and Apr 2006.ESBLs were tested by phenotypic confirmatory tests recommended by CLSI.Twenty-one kinds of ?-lactamases genes of blaTEM,blaSHV,blaLEN,blaOKP,blaCTX-M-1 group,blaCTX-M-2 group,blaCTX-M-9 group,blaOXA-1 group,blaOXA-2 group,blaOXA-10 group,blCARB,blaPER,blaVEB,blaGES,blaLAP,blaDHA,blaACT/MIR,blaCMY/MOX,blaFOX,blaCMY/LAT,and blaACC were analyzed by PCR and verified by DNA sequencing.RESULTS In 25 strains of K.pneumoniae,the positive,negative,and "uncertainty" rates of ESBLs were 56.0%,20.0%,and 24.0%,respectively.The positive rate of genes of blaTEM,blaSHV,blaCTX-M-1 group,blaOXA-10 group,blaLAP,and blaDHA were 80.0%,4.0%,4.0%,80.0%,4.0% and 32.0%,respectively.The 15 kinds of rest genes were all tested negative.The total positive rate of 21 kinds of ?-lactamases gene was 92.0%.Among them,the blaLAP-2 gene sequence of the HZ12593 strain has been registered in GenBank(GenBank Accession Number: EU529981).CONCLUSIONS There are higher rate of ESBLs-producing strains in K.pneumoniae isolated from the inpatients,and at least 6 kinds of ?-lactamases gene existed.Both genes of blaTEM and blaOXA-10 group are the most common genotypes.Carring blaDHA Gene may influence the result of phenotypic confirmatory test for ESBLs in K.pneumoniae.

Objective To investigate the transcriptional autoregulation of PhoP/PhoQ under different growth conditions in Yersinia pestis.Methods The entire promoter region of YPO1635 was amplified and cloned into the pRW50 vector containing a promoterless lacZ reporter gene.The recombinant LacZ reporter plasmid was transformed into the wild-type strain (WT) and the phoP mutant strain (ΔphoP),respectively,to measure the promoter activity (the β-galactosidase activity) of the target gene in WT and ΔphoP by using the β-galactosidase enzyme assay system.Total RNAs were extracted from WT and ΔphoP strains,and primer extension assay was employed to detect the promoter activity by examining the amount of primer extension products of YPO1635 in WT and ΔphoP.Results The LacZ fusion results showed that the transcription of YPO1635 was positively regulated by PhoP under L-TMH and brain-heart infusion(BHI) conditions,but it was not regulated in H-TMH medium.The primer extension assay detected two transcriptional start sites located at 90 and 118 bp upstream of the translation initiation site of phoP,named P1 and P2,respectively.Under low Mg2+ TMH conditions,the promoter activity of P1 rather than P2 was positively regulated by PhoP.Under high Mg2+ TMH conditions,the promoter activities of both P1 and P2 showed no obvious difference in the WT and ΔphoP strains.Under rich BHI conditions,both promoters were under negative control of PhoP.Conclusion Different autoregualtion patterns of PhoP/PhoQ under different growth conditions would help Y.pestis to quickly adapt to the changing living environment.

Objective To study the transcriptional regulation of vp1667 by H-NS in Vibrio parahaemolyticus.Methods Total RNAs were extracted from Δhns and WT strains.Quantitative RT-PCR was carried out to calculate the transcriptional variation of vp1667 between Δhns and WT.Primer extension assay was also employed to detect the transcription start site and the promoter activity (i.e.the amount of primer extension products) of vp1667 in Δhns and that in WT.The promoter DNA region of vp1667 was amplified, purified, and cloned into the corresponding restriction endonuclease sites of pHRP309 that harbors a gentamicin resistance gene and a promoterless lacZ reporter gene.The recombinant pHRP309 plasmid was transformed into Δhns and WT, respectively, while β-galactosidase activity in cellular extracts was measured using a β-galactosidase enzyme assay system.The over-expressed His-H-NS was purified under native conditions with nickel loaded HiTrap Chelating Sepharose columns.The electrophoretic mobility shift assay (EMSA) and DNaseⅠ footprinting were then applied to analyze the DNA-binding activity of His-H-NS to vp1667 promoter region in vitro.Results and Conclusion The primer extension assay detected one transcription start site for vp1667, which was located at 28 bp upstream of vp1667, and its transcribed activity was under the negative control of the H-NS.The EMSA and DNaseⅠ footprinting assay results showed that His-H-NS was unable to bind to the promoter-proximal DNA region of vp1667, suggesting that H-NS indirectly inhibits the transcription of vp1667.

Objective To evaluate the relationship between serum tissue inhibitors of metaUoproteinase-1 and children with myocarditis,and provide evidences to support the measurement of serum tissue inhibitors of metallopro- teinase-1. Methods Tissue inhibitors of metalloproteinaee-1 and bioehem indexes of the harvested specimens and e -chocardiogram were measured from children with myocarditis(age: 1～14 years old, n = 9) and healthy young chil- dren(age: 1～14 years old, n = 9). Results Children with myocarditis showed reduced ejection fraction and dilated left ventricular compared with healthy children. Tissue inhibitors of metalloproteinaee-1 in children with myocarditis was significantly reduced compared with healthy children. Aspartie acid transanainase, lactate dehydrogenase,creatine kinase, hydroxybutyrie acid dehydrogenase, MB isoenzyrne of creatine kinase, cardiac troponin Ⅰ , C reactive protein showed no difference between two groups. Conclusion The reduced serum level of tissue inhibitors of metallopro- teinase-1 may show the severity of children with myocarditis and maybe one of the mechanisms of children with di- lated cardiomyopathy following myocarditis.

Objective To compare the differences of hospital stays,hospitalization costs and effectiveness via keyhole hematoma debridement (KHED) and internal medicine conservative treatment (IMCT) in treating 30-40 mL hypertensive intracerebral hemorrhage.Methods Fifty-eight patients with hypertensive intracerebral hemorrhage whose bleeding was 30-40 mL,admitted to our hospital from January 2014 to September 2015,were chosen in our study;according to the will of the patients and their family members,the patients were divided into KHED group (n=31) and IMCT group (n=27).The differences of hospital stays,hospitalization costs,neurological dysfunction rate at hospital and three months after discharge,and recovery results were compared between the two groups.Results The average hospital stays of KHED group were (7.4±2.3) d and those of IMCT group were (14.5±5.1) d,with significant difference (P=0.012);the hospitalization costs for the two groups were (36 296.28±5292.12)yuan,and (41 769.48±6342.83) yuan,with significant difference (P=0.027).Glasgow outcome scale of KHED group at discharge indicated 29 patients with good recovery and 2 with poor recovery;that of IMCT group indicated 20 with good recovery and 7 with poor recovery,including two with cerebral edema accepted craniotomy operation in second time.Follow-up for three months showed that the KHED group had basic activities of daily living in 16 patients,mild hemiplegia in 11 and severe hemiplegia in 4,and IMCT group had basic activities of daily living in 9 patietns,mild hemiplegia in 13 and severe hemiplegia in 5;significant differences were noted between the two groups (Z=2.499,P=0.001).Conclusion KHED in treatment of 30-40 mL hypertensive intracerebral hemorrhage can shorten hospitalization time,reduce cost,have better prognosis and better short-term and long-term effectiveness than IMCT.