Objective To investigate the effect of fucosyltransferase 7(Fut7) dysregulation on lipopolysaccharide(LPS)-stimulated inflammatory response in human monocytes THP-1, so as to provide experimental evidence for the study of inflammatory induction mechanism of clinical related diseases.Methods THP-1 cells were treated with different concentrations of LPS(0, 0. 5, 1, 2, 4, 10 μg/mL). The transcription levels of interleukin-1β(IL-1β), IL-6, IL-8 and tumor necrosis factor-α(TNF-α) in the cells were detected by RT-qPCR, and the LPS stimulation concentration for establishing inflammatory cell model was determined. THP-1 cells were stimulated with the optimum concentration of LPS to establish the inflammatory cell model. The effects of Fut7, sialyl Lewis X(sLeX) and Fut7 activity inhibitor SGN-2FF on the transcription levels of IL-1β, IL-6,IL-8 and TNF-α genes in inflammatory cell model were detected by RT-qPCR.Results The LPS stimulation concentration for establishing inflammatory cell model was 4 μg/mL. Overexpression of Fut7 and sLeX significantly increased the transcription levels of IL-1β, IL-6, IL-8 and TNF-α in LPS-induced THP-1 cell inflammatory model(t = 3. 93-30. 05, each P < 0. 05).Inhibiting the expression of Fut7 significantly decreased the transcription levels of IL-1β, IL-6, IL-8 and TNF-α in the inflammatory cell model(t = 6. 98-39. 50, each P < 0. 01). SGN-2FF significantly decreased the transcription levels of IL-1β, IL-6and IL-8(t = 5. 17-6. 44, each P < 0. 001), while significantly increased the transcription level of TNF-α(t = 12. 31, P < 0. 001).Conclusion Overexpression of FuT7 and sLeX can enhance the LPS-induced inflammatory response in THP-1 cells by promoting the expression of inflammatory cytokines. Inhibiting the expression of Fut7 and SGN-2FF, an inhibitor of Fut7enzyme activity, can effectively relieve inflammation, which provides a new strategy for the treatment of inflammatory diseases.

Objective To investigate the possibility of differentiation of multipotent adult progenitor cells from human bone marrow(hMAPCs) into hepatocytes with hepatocyte growth factor (HGF) and fibroblast growth factor-4 (FGF-4) in vitro to find a new cell source for liver tissue engineering. Methods (1) Harvest of the hMAPCs: The bone marrow mononuclear cells were obtained from bone marrow obtained from volunteers by gradient centrifuging. The adherent mononuclear cells were cultured. The mesenchymal stem cells(MSCs) were isolated and collected by magnetic activated cell sorting(MACS) through depletion selection by the use of CD45 and GlyA microbeads. (2) The hMAPCs were induced to differentiate with HGF(20ng/ml) +FGF-4(10ng/ml). L-02 human hepatocytes (cell lines) were used as the control, with undifferentiated hMAPCs serving as the contrast. (3)The expression of albumin (ALB), alpha fetoprotein (AFP), cytokeratin-18(CK-18), cytokeratin-19(CK-19) was detected with immunocytochemistry to identify the characters of the differentiated cells at different time, and to determine the ratio of positive cells. (4) ALB expression of the differentiated cells at different time was detected by Western blot on day 21 and 25. Results (1) The result of immunocytochemistry: The differentiated hMAPCs expressed the late markers (ALB, CK-18) of hepatocyte at all the time but did not express CK-19. The differentiated hMAPCs expressed AFP on day 7. (2) The result of Western blot assay: the differentiated cells expressed the late markers (ALB) of hepatocyte on day 21 and 35. Conclusions hMAPCs can differentiate into hepatocyte-like cells under specific inducing conditions.

Objective:To inve tigate the value o electron beam tomography and echocardiography or diagno ing the comple congenital heart di ea e . Method :Thirty our patient with comple congenital heart di ea e underwent both echocardiography and electron beam tomography,which wa con irmed by angiocardiography in 18 patient .Twenty one patient were operated on.All data were comparatively analyzed Re ult :The rate o diagno i coincidence in diagno ing comple congenital heart di ea e wa 82 4% or electron beam tomography,and 73 5% or echocardiography.The accuracy o electron beam tomography in diagno ing e tracardiac abnormal tructure wa 97 9% and obviou ly higher than 53 2% by echocardiography.The accuracy o echocardiography in diagno ing intracardiac abnormal tructure wa 95 9% and higher than 81 6% by electron beam tomography.Electron beam tomography and echocardiography had a imilar accuracy or diagno ing abnormal tructure at the junction o ve el with cardiac chamber. Conclu ion:Electron beam tomography combined with echocardiography can greatly improve the diagno i accuracy o comple congenital heart di ea e and can al o reduce the u e o inva ive angiocardiography.

0.05), but there were significant differesnces compared with the control group ( P

Ikaros is a regulatory factor playing an important role in control of the development of hema-topoietic cells and immune system;and as a tumor suppressor,its aberration can cause both T and B lineage development arrest and neoplastic transformation,leading to insensitive to therapy and poor prognosis in actue lymphoblastic leukemia. Recently,researches on how Ikaros affects the leukemogenesis and prognosis become the focus of attention.

Objective To observe the expression of retinol-binding protein 4 (RBP4) and phosphatidylinositol 3 kinase (PI3K) in insulin resistant (IR) rats and the role of pioglitazone intervention. MethodsThirty-five male Wistar rats in SPF level were randomly divided into control fed with normal diet (n= 11 ) and IR model fed with high fat sucrose diet (HFSD) (n= 24). The IR rats were then randomly divided into two subgroups, namely IR fed with HFSD(n = 12) and pioglitazone intervention (n= 12) given a daily dose of 20 mg · kg 1 · d-1 pioglitazone and HFSD (IR +Pio group )for 8 weeks.All rats were killed after 16 weeks and the levels of serum TG, high density lipoprotein (HDL-C), low density lipoprotein (LDL-C), fasting blood-glucose (FBG), fasting serum insulin (FIns) and insulin resistance index (HOME-1R) were measured.Enzyme-linked immunosorbent assay (ELISA) and RT-PCR were employed to detect the level of serum RBP4 and the mRNA expression of RBP4 in the epididymis adipose tissues, respectively. Immunohistochemistry was used to detect the protein expression of PI3K in skeletal muscles while UV spectrophotometer was employed to measure level of serum AOPP. The ratio of visceral fat weight of mesentery, epididymis and peritoneum in abdominal cavity to body weight (BW) was calculated. Results(1) the level of BW, TG, LDL-C, FINS and the ratio of visceral fat weight to BW were higher in IR group than in control group, but HDL-C decreased significantly. After the intervention of pioglitazone, the level of BW,TG, LDL-C, FINS, and the ratio of visceral fat weight to BW in IR+Pio group were lower and HDL-C increased significantly than those in IR group.(2) The level of RBP4 from serum and epididymis adipose and serum AOPP were higher in IR group than in control and lower significantly in IR+Pio group than in IR group (all P＜0.05). (3) The expression of PI3K in skeletal muscles were lower in IR group than in NC group, and increased after the invention of pioglitazone. (4) FIns, the ratio of visceral fat weight to BW and LDL-C were positively correlated with RBP4 while HDL-C and PI3K negatively correlated with RBP4. Conclusions(1) The increased RBP4 can lead to metabolic disturbances and oxidative stress in IR rats.(2) RBP4 may decrease the insulin sensibility by weakening insulin signal transduction. (3) Pioglitazone can ameliorate insulin resistance by decreasing the level of RBP4 and serum AOPP and increasing the level of PI3K in skeletal muscles of IR rats.

Preoperative portal vein embolization(PVE)has become an important tool in the management of selected patients with hepatic cancer before the major hepatic resection is carried out.PVE can redirect the portal flow to the intended future remnant liver tissue in order to induce the hypertrophy of the non-diseased portion of the liver and thereby may reduce the occurrence of complications and shorten the hospitalization days after surgery.This article aims to review the technical and clinical considerations in performing PVE and to discuss the PVE-related practical points,including the relevant anatomy,the access approach,the choosing of embolic agents and the pathophysioiogy of PVE.In addition,the indications and contraindications for performing PVE,the use of combination therapies and the concern for tumor growth after PVE are also discussed.

Objective To investigate the prevalence, molecular mechanism and genetic characteristics of azithromycin-resistant Neisseria gonorrhoeae (N.gonorrhoeae) strains isolated in Shenzhen.Methods N.gonorrhoeae strains were collected in Shenzhen from 2011 to 2015.Agar dilution method and E-test were used to detect the minimum inhibitory concentrations (MIC) of these strains to azithromycin.All azithromycin-resistant (AZM-R) strains (MIC≥2 μg/ml) and some azithromycin-sensitive strains (MIC≤0.25 μg/ml) which were randomly selected as the control group were screened for mutations in 23S rRNA, mtrR and erm genes and genotyped by using N.gonorrhoeae multi-antigen sequence typing (NG-MAST).Results A total of 788 N.gonorrhoeae strains were collected, 148 (18.8%) of which were AZM-R strains (MIC≥1 μg/ml).Eighteen out of 21 high-level AZM-R (AZM-HLR) strains had A2143G mutations in the four copies of the 23S rRNA gene.Twelve out of 29 middle-level AZM-R (AZ-MLR) strains had missense mutations, among which C2611T mutations in the four copies of the 23S rRNA were detected in 10 strains.Incidence of G45D/Y105H mutation in AZM-HLR strains was higher than that in AZM-MLR (χ2=12.702, P=0.000) or AZ-S (χ2=4.462, P=0.035) strains according to the analysis of the promoter and coding region of mtrR gene.PCR analysis revealed that only one strain carried ermB gene (MIC=2 μg/ml).The 788 N.gonorrhoeae strains were typed into 81 sequence types (STs) by NG-MAST, most of which were represented by one strain only.STs of ST3356 and ST1866 that were identified in the AZ-R strains in the current study had been noted in a previous report of emerging AZM-R N.gonorrhoeae strains in Nanjing, Chongqing and Guangzhou.Neighbor-joining (NJ) phylogenetic tree showed that the resistant strains did not form a separate cluster.Conclusion Currently, it is not suitable to use azithromycin as a monotherapy for gonorrhea in Shenzhen.Mutations of A2059G and C2611T in 23S rRNA of N.gonorrhoeae were respectively responsible for high-level and middle-level resistance to azithromycin.Repeated emergence of ST1866 and ST3356 will help us monitor and analyze the epidemiological characteristics of N.gonorrhoeae strains resistant to azithromycin in Shenzhen.

Objective To establish a real‐time quantitative PCR method for the detection of cytokine receptor‐like factor 2 (CRLF2) expression .Methods Specific primers amplification target gene CRLF2 and housekeeping genes ABL were designed ,the purified PCR products were performed the TA cloning .After bacterial colony PCR screening and sequencing ,then the recombinant plasmids DNA was extracted and measured by using UV spectrophotometer and converted to copies/mL concentration .Finally it was diluted for preparing the plasmid standard substance ,then the standard curve was drawn for observing the sensitivity and linear rang ,meanwhile the stability of the plasmid DNA was evaluated .This method was initially applied to detect the CRLF2 level of bone marrow mononuclear cells in 10 cases of healthy children and 10 cases of newly diagnosed acute lymphoblastic leukemia (ALL) .Results CRLF2 PCR product had a single specific melting curve;the linear detection range of the standard substance was 103 - 108 copies /ml;the plasmid standard substance by freeze‐thawing for 3 times remained stable;the CRLF2 level of clinical sample was within the linear detection range of standard substance .Conclusion The real‐time quantitative PCR method for CRLF2 established by our laboratory has good specificity ,linearity range and stability ,which can be applied to the quantitative detection of CRLF2 gene in clinical ALL children .

AIM: To study the effect of moderate hypothermia on immature rats with hypoxic-ischimic brain damage (HIBD). METHODS: The rats with HIBD were divided into normothermic recovered group (IN) and moderate-hypothermic recovered group (IH). Sham-operated rat pups were normothermic control group (NC) and moderate-hypothermic control group (HC). 0, 2, 6, 24, 48, 72 h after the end of hypoxic-ischemic (HI) insult, the brain was homogenized for measuring glucose and ATP, brain mitochondria was extracted for SDH activity, complex II activity and the capacity of ATP synthesization. RESULTS: In IN group, the brain glucose was significantly lower at 0 h, and recovered as normal at 2 h. The brain ATP and brain-mitochondrial SDH activities were firstly decreased at 2 h, 6 h and then recovered gradually, it was at it's peak value at 72 h. Brain-mitochondrial complex II activity and the capacity of ATP synthesis were recovered at 2 h, but they decreased again at 6 h and came to normal level at 72 h. In moderate-hypothermic group, all the indexes were significantly higher at all the time point than that in IN groups. CONCLUSION: Moderate hypothermia inhibits the decrease in the mitochondrial SDH activity, mitochondrial complex II activity and the capacity of ATP synthesis, increases the brain ATP concentration, improves the energy metabolism, and then protects the brain tissue. [