Study on value of HbA1c,GSP,SHBG,TG and FFA for diagnosing gestational diabetes mellitus / 国际检验医学杂志

Objective To investigate the clinical value of glycosylated hemoglobin(HbA1c),glycosylated serum protein(GSP),sex hormone binding globulin(SHBG),three acylglycerol(TG),free fatty acid(FFA)in diagnosing gestational diabetes mellitus(GDM).Methods 103 cases of GDM patients and 98 cases of healthy pregnant women from February 2015 to August 2016 were selected as the GDM group and control group.The positive detection rates and levels of HbA1c,GSP,SHBG,TG and FFA were compared between the two groups.Moreover the diagnostic efficiency of various indicators was analyzed.Results The levels of HbA1c, GSP,TG and FFA in the GDM group were significantly higher than those in the control group,while the SH-BG level was significantly lower than that in the control group,the difference was statistically significant(P<0.05);the positive rates of HbA1c,GSP,SHBG,TG and FFA in the GDM group were significantly higher than those in the control group,the difference was statistically significant(P<0.05);the specificity,sensitivity and positive prediction value of HbA1c,GSP,SHBG,TG and FFA for jointly diagnosing GDM were signifi-cantly higher than those of single indicator,the difference was statistically significant(P<0.05).Conclusion Detecting HbA1c,NGSP,SHBG,TG and FFA is more accurate for jointly diagnosing GDM,has an important diagnostic value,and can serve as the assisted diagnostic indicators.

HIV small-molecule entry inhibitors targeting gp120, CD4 and CXCR4:research advances / 国际药学研究杂志

AIDS is an infectious disease caused by human immunodeficiency virus(HIV)and has done great harm to human beings. The envelope glycoprotein surface subunit gp120 and its receptor CD4 and coreceptor CXCR4 play important roles in HIV-1 en?try. The Phe43 pocket and Arg59 of gp120 are two important regions that interact with CD4. Recently,HIV small-molecule entry inhib?itors have become a hot topic in anti-HIV drug research,which are able to inhibit the virus before the cells are infected. The Phe43 pocket is an attractive target and the study of Phe43 pocket-targeting small-molecule entry inhibitors is underway,including BMS-378806,BHS-488043 and its analogs,and NBD-556 and its analogs. Besides,CXCR4 antagonist is another important approach.

Constructing CXCR4 lentiviral vector and transfecting breast cancer cells / 实用医学杂志

Objective To construct lentivirus containing CXCR4 gene and transfect MCF-7 cells , and obtain CXCR4 high-expressing MCF-7 cells. Methods CXCR4 gene was amplified by RT-PCR to construct CXCR4/pSin-EF2, which was transfected into HEK293T cells with psPAX2 and pMD2G vector for lentivirus packing. Packaged lentivirus was used to transfect human breast cancer cells MCF-7, with empty lentivirus as control. CXCR4 mRNA and protein expression levels were detected by RT-PCR and Western blot before and after transfection. And flow cytometry was used to detecte cell surface CXCR4 expression. Results The recombinant plasmid CXCR4/pSin-EF2 was constructed successfully,identified by double digestion and sequencing, and transfected into HEK293T cells to obtain high-titer lentivirus. RT-PCR and Western blot confirmed that the expression of CXCR4 in MCF-7 cells increased significantly after CXCR4 lentivirus transfection. Flow cytometry results showed that the CXCR4 positive rate increased from 26.78% to 99.29%, while there is no significant difference in CXCR4 expression between vector-transfected MCF-7 cells and non-transfected MCF-7 cells. Conclusion CXCR4 lentivirus and the breast cancer cell line with high and stable expression of CXCR4 (MCF-7CXCR4) were successfully constructed.

Soluble expression and activity evaluation of SDF-1/54R, a specific antagonist of CXCR7 / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a soluble prokaryotic expression vector of the CXCR7-specific antagonist SDF-1/54R and evaluate its activity.</p><p><b>METHODS</b>SDF-1/54r gene amplified by PCR was inserted into the soluble expression vector pET-41a+ engineered with GST fusion tag, and the recombinant vector was transformed into E. coli strain BL21 (DE3). After IPTG induction of E. coli, the expressed recombinant protein was purified with GST affinity chromatography purification system and confirmed by SDS-PAGE and Western blotting assay. The target protein SDF-1/54R was obtained after digestion of the purified product with enterokinase. Breast cancer MCF-7 cells with high expression of CXCR7 was treated with SDF-1/54R and the cell proliferation and metastasis was evaluated with MTT and chemotaxis assays.</p><p><b>RESULTS</b>The target protein SDF-1/54R obtained showed an obvious inhibitory effect on the proliferation and metastasis of MCF-7 cells as confirmed by MTT and chemotaxis assays.</p><p><b>CONCLUSION</b>SDF-1/54R is a good antagonist of CXCR7 and shows a potential value as an effective anti-cancer agent.</p>

Construction of SDF-1P2G54, a specific antagonist of CXCR4 / 南方医科大学学报

<p><b>OBJECTIVE</b>To obtain a specific antagonist of CXCR4, SDF-1P2G54 by mutating SDF-1 second proline (P) into glycin (G) and removing the α-helix of its C-terminal.</p><p><b>METHODS</b>SDF-1p2g54 gene amplified by PCR was inserted into the vector pET-30a (+) and transformed into Escherichia coli (E. coli) strain BL21. After IPTG induction of E. coli, the expressed recombinant protein was purified with nickel-affinity chromatography column under denaturing conditions and refolded with gradient dilution and ultra-filtration. The chemotactic effect of SDF-1P2G54 on Jurkat cells and its antagonistic effect against SDF-1 were determined by transwell assay; flow cytometry was used to assay the ability of SDF-1P2G54 to induce calcium influx and CXCR4 internalization in MOLT4 cells.</p><p><b>RESULTS</b>The recombinant protein SDF-1P2G54 completely lost the functions to activate CXCR4 or to induce transmembrane migration of Jurkat cells and calcium influx in MOLT4 cells, but maintained a high affinity to CXCR4. SDF-1P2G54 effectively inhibited the chemotactic effect of wild-type SDF-1 to Jurkat cells, and induced rapid CXCR4 internalization in MOLT4 cells.</p><p><b>CONCLUSION</b>SDF-1P2G54 is a new antagonist of CXCR4 with a potential value as an effective inhibitor of HIV-1 infection, cancer metastasis or other major diseases.</p>

Construction of intrakine mutant SDF-1alpha/54/KDEL and its inhibiting effects upon CXCR4 expression on cell membrane / 生物医学工程学杂志

To investigate the impact of phenotypic knockout of CXCR4 on Molt-4 cells via intrakine technology,the C-terminal alpha-helix gene SDF-1alpha/54/KDEL of human stromal cell-derived Faceor-1 deletion is fused to a retention signal 4-peptide -KDEL that retains the newly synthesized receptor within the Molt-4 cells endoplasimc reticulum. Subsequently, PCR is used to amplify the target gene SDF-1alpha/54/ KDEL from the constructed plasmid SDF-WT-Gly x 4-Dec/PET-30a(+) at its C-terminal and subclone it into eukaryotic expression vectors pEGFP-C3 for generating recombinant vector cells by lipEGFP-C3/SDF-1alpha/54/KDEL, and then have it sequenced. After the transfection of recombinant plasmids into COS-7 posome, SDF-1alpha/54/KDEL protein is confirmed with Western blot. The recombinant plasmids pEGFP-C3/SDF-1alpha/54/KDEL are isolated and transiently transfected in Molt-4 cells by electroporation. Flow cytometric analysis shows a dramatic reduction of CXCR4 expression on Molt-4 cells. The conclusion is that SDF-1alpha/54/KDEL could assume a role in the phenotypic knockout of CXCR4, and the findings suggest that the inhibiting effect of SDF-1alpha/54 against CXCR4 is not influenced by the deletion of SDF-1alpha helix at the C terminal.

The action of decorin in anti-fibrosis and anti-cancer / 生物医学工程学杂志

Decorin (DCN) is a member of the small leucine-rich proteoglycan gene family. Many studies indicated that DCN inhibited fibrosis and scar-formation by neutralization of TGF-P and interfering the binding of TGF-beta with its receptor, which induced ectopic deposition of extracellular matrix. Additionally, DCN can prevent the proliferation and metastasis of tumor cells by activating EGFR/MAPK/p21 signal pathway and inhibiting the cell proliferation pathway mediated by EGF-EGFR. It is suggested that the recombinant DCN had potential pharmaceutical potency in treatment of chronic fibrosis and neoplasm for its critical biological activities and low immunogenicity.

The role of SDF-1 and its receptor CXCR4 in tumor metastasis / 生物医学工程学杂志

The chemokine SDF-1 (CXCL12) and its receptor, CXCR4, have been implicated in organ-specific metastases of several malignancies. CXCR4 expression has recently been characterized in many cancer cell types and is thought to play a pivotal role in directing the migration of metastasizing tumor cells to SDF-1-rich tissues. SDF-1, which is highly expressed in the organs where breast cancers preferentially metastasize, has been shown to promote cancer cell migration. The tumor cells use chemotaxis which occurred between CXCR4 and its ligand SDF-1 to direct migration from their primary sites via the circulation to the preferential sites of metastases, and further studies on the mechanism involved in a variety of cellular signaling pathways are beneficial to tumor therapy.

Extraction and Isolation of Different Polar Alkaloids from Herba Dendrobii and Their Influences on Human Lens Epithelial Cells Proliferation / 广州中医药大学学报

Objective To observe the protection of human lens epithelial cells(HLEC) by different polar alkaloids extracted from Herba Dendrobii(HD).Methods We extacted the Herba Dendrobii powder with ethanol,and then treated the extract with falling-film concentration,acidification,salting out,chloroform extraction,and washing with water.Different polar alkaloids were extracted from HD after the above treatment.The protective effect of HD alkaloids was observed on HLEC,which were cultured with DMEM medium containing 10% fetal calf serum.Ten groups were set up for the experiment: normal control group,model group,high-and low-dose water-soluble alkaloids groups,high-and low-dose fat-soluble alkaloids groups,high-and low-dose low-polar alkaloids group,and high-and low-dose weak-polar alkaloids groups.The high-dose dosage of the alkaloids was 25.0 ?g/L and low-dose dosage was 12.5 ?g/L.Methyl thiazolyl tetrazolium(MTT) assay was used to evaluate the proliferation of HLEC under the different conditions of interventions.Results The single-factor experiments showed that the highest extracting rate of HD alkaloids was obtained under the conditions of extracting the powder with 80% ethanol for 3 times and for 3 hours.The results of protective experiment showed that the proliferation of HLEC in the model group was inhibited by hydrogen peroxide(H2O2),and the inhibitive rate was lower in low-dose fat-soluble alkaloids group than that in the model group(P