Traditional herbal medicines, Panax ginseng, Panax quinquefolium and Panax notoginseng, attract our attention for their extensive and powerful pharmacological activities. Ginsenosides are the main active constituents of these medicinal herbs. The related glycosyltransferases involved in ginsenoside biosynthesis are the key enzymes which catalyze the last important step. Modification of ginsenoside aglycones by glycosyltransferases produces the complexity and diversity of ginsenosides, which have more extensive pharmacological activity. At present, ginsenoside aglycones and compound K have been obtained by synthetic biology. As the last step of ginsenoside biosynthesis, glycosylation of ginsenoside aglycones has been studied intensively in recent years. This review summarizes the basic strategies and research advances in studies on glycosyltransferases involved in ginsenoside biosynthesis, which is expected to lay the theoretical foundation for the in-depth research of biosynthetic pathway of ginsenosides and their production by synthetic biology.
Biosynthetic Pathways ; Ginsenosides ; biosynthesis ; Glycosyltransferases ; metabolism ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Synthetic Biology

Objective To study the effects of ?2-adrenergic receptor gene Arg16Gly polymorphism on blood lipid and apolipoprotein ratio and its role in blood lipid and apolipoprotein ratio mediated by high carbohydrate/low fat (HC/LF) diet in healthy young persons.Methods Fifty-six healthy young volunteers had regular diet for 7 d followed by HC/LF diet for 6 d.Twelve-hour fasting venous blood samples were collected on days 1,8 and 14 to measure blood lipid and apolipoprotein (apo) AI and B100 levels,and to calculate ratios of TG/HDL-C,log (TG/HDL-C),TC/HDL-C,LDL-C/HDL-C and apoAI/apoB100.DNA was isolated from genome.Arg16Gly polymorphism was analyzed by PCR-RFLP.Results No significant difference was found in the baseline lipid and apolipoprotein ratio in subjects with AA genotype and G carriers before and after regular or HC/LF diet.The ratios of TG/HDL-C (P=0.017),log (TG/HDL-C) (P=0.031),and apoAI/apoB100 (P=0.006) were significantly higher,while those of TC/HDL-C (P=0.001) and LDL-C/HDL-C (P

Objective:To investigate the expression of miR-92a in colorectal cancer (CRC) and its effect on the regulation mechanisms of tumor angiogenesis. Methods:The miRNA-92a expression in 25 CRC tissues and HT29, SW620, SW480, and HCT116 CRC cells was detected using quantitative real-time polymerase chain reaction. The CD31 positive expression of blood microvessels in CRC tissues was measured by immunohistochemistry. Pearson correlation analysis was used to analyze the relationship between miR-92a and tu-mor angiogenesis. The miR-92a mimic or inhibitor was transfected into HCT116 and SW620 cells in vitro to upregulate or downregu-late the miR-92a expression. The effect of miR-92a overexpression or suppression on the formation of human umbilical vein endotheli-al cell (HUVEC) tubules was detected by tube formation assay. Changes in phosphatase and tensin homolog (PTEN) protein expression were measured by Western blot. Results:The expression level of miR-92a in CRC tissues was significantly higher than that in matched adjacent tissues (P<0.01). The expression levels of miR-92a in HT29, SW480, SW620, and HCT116 colon cancer cell lines were signifi-cantly higher than that of the normal colorectal epithelium control (P<0.05). The number of CD31 positive expression of microvessel density (MVD) in CRC tissues was significantly higher than that in adjacent tissues (P<0.01), and the miR-92a expression level was posi-tively correlated with the MVD in CRC tissues (r=0.580, P=0.01). The cell culture supernatant of HCT116 with miR-92a overexpression could significantly promote the formation of HUVEC tubules (P<0.05). The upregulation of miR-92a expression could significantly inhib-it the expression of PTEN protein in CRC cells (P<0.01). Conclusion:The miR-92a was highly expressed in CRC cells and tissues, which was closely related to the formation of tumor angiogenesis in CRC. The miR-92a could promote tumor angiogenesis in CRC by inhibit-ing PTEN expression.

Objective To explore the expression characteristic of serum mir-16,mir-21,mir-29a, mir-92a and mir-143 in colorectal cancer , evaluate the clinical significance of candidate miRNAs in CRC diagnosis.Methods Case-control study was used.The expression levels of serum mir-16,mir-21,mir-29a, mir-92a and mir-143 from 50 CRC patients and 27 normal controls were detected by real-time PCR, the levels of miRNAs expression in serum of 8 CRC patients 1 day before and 7 days after radical surgery were analyzed.The sensitivity and specificity of serum miRNAs expression for the diagnosis of colorectal cancer were evaluated using the receiver operating characteristic ( ROC) curve.Results The expression level of mir-16, miR-21, mir-92a in serum of patients with CRC were (75.55 ±37.73), (35.96 ±23.81), (24.79 ±8.97) fmol/ml, significantly higher than that of the healthy control group (32.73 ±18.94), (24.36 ±13.27), (16.36 ±5.58) fmol/ml (tmir-16 =2.77, tmir-21 =2.34, tmir-92a =3.85,P <0.05).However, the expression levels of mir-29a and mir-143 showed no significant difference between two groups (tmir-29a=-0.17, tmir-143 =1.59,P>0.05).In addition, serum miR-16 and miR-92a levels of CRC patients 7 days after tumor resection were (36.02 ±19.95), (14.82 ±7.78) fmol/ml, significantly lower than that before operation (62.18 ±34.17), (24.06 ±12.99) fmol/ml (tmir-16 =3.59, tmir-92a =2.60,P<0.05).Area under the ROC curve ( AUC) of combined detection of mir-16, miR-21and mir-92a was 0.877, the sensitivity and specificity were 88% and 85%, respectively, which was higher than any of 3 miRNAs alone and the conventional tumor marker CEA.Conclusion Combination of mir-16, mir-21 and mir-92a in the diagnosis of CRC shows a better sensitivity and specificity , which would be expected as new tumor biomarkers for noninvasive diagnosis and monitoring progression of colorectal cancer.

The cyclization of 2,3-oxidosqualene is the key branch point of ergosterol and triterpenoid biosynthesis. Downregulation of 2,3-oxidosqualene metabolic flux to ergosterol in Saccharomyces cerevisiae may redirect the metabolic flux toward the triterpenoid synthetic pathway. In our study, primers were designed according to erg7 gene sequence of S. cerevisiae. Three fragments including 5' long fragment, 5' short fragment and erg7 coding region fragment were amplified by PCR. 5' long fragment consists of the promoter and a part of erg7 coding region sequence. 5' short fragment consists of a part of promoter and a part of erg7 coding region sequence. These fragments were inserted reversely into pESC-URA to construct antisense expression plasmids. The recombinant plasmids were transformed into S. cerevisiae INVSc1 and recombinant strains were screened on the nutritional deficient medium SD-URA. The erg7 expression level of recombinant strains, which harbored antisense expression plasmid of erg7 coding region, was similar to that of INVScl by semi-quantitative PCR detection. But erg7 expression level of recombinant strains, which harbored 5' long antisense fragment and 5' short antisense fragment, was significantly lower than that of the control. The results of TLC and HPLC showed that the ergosterol content of recombinant strains, which harbored 5' long antisense fragment, decreased obviously. The ergosterol contents of the others were almost equal to that of INVSc1. Lanosterol synthase gene expression was downregulated by antisense RNA technology in S. cerevisiae, which lays a foundation for reconstructing triterpenoid metabolic pathway in S. cerevisiae by synthetic biology technology.
DNA Primers ; Down-Regulation ; Gene Expression ; Intramolecular Transferases ; genetics ; metabolism ; Plasmids ; Polymerase Chain Reaction ; RNA, Antisense ; Saccharomyces cerevisiae ; enzymology ; genetics ; Squalene ; analogs & derivatives ; metabolism ; Transformation, Genetic

Objective To evaluate whether S-100β protein could be a serum marker for traumatic spinal cord injury (SCI). Methods From June, 2013 to October, 2014, 24 patients with complete SCI were measured the serum S-100β protein concentrations with en-zyme-linked immunosorbent assay, one week, three and six months after SCI. Serum from ten healthy persons was as normal control. Re-sults The serum S-100βprotein concentrations increased one week and 3 months after SCI (Z>4.273, P<0.001). Conclusion The increase of serum S-100βprotein may help assessing early impairment after complete SCI.

The mechanism of injury on the human glomerular endothelial cells (ciGENC) induced by preeclampsia serum was investigated. Concentration of maternal serum sFlt-1 protein was detected by ELISA. Fluorescently-labeled bovine serum albumin infiltrating through lower chamber of Transwell was measured by multifunction microplate reader. Morphologic change of ciGENC was observed under inverted phase contrast microscope. The concentration of sflt-1 in preeclampsia groups was significantly increased as compared with control group (P<0.01). Permeability in preeclampsia groups was significantly increased as compared with control group (P<0.01). By contrast with severe preeclampsia group, the permeability of ciGENC monolayer in mild preeclampsia group was decreased significantly (P<0.05). Intervention of exogenous VEGF significantly decreased permeability of ciGENC in preeclampsia groups. It was concluded that sFlt-1 increased ciGENC permeability by damaging integrity of endothelial barrier function.

This study examined the effect of over-expression of sFlt-1 by trophoblasts on the barrier function of glomerular endothelial cells and the role of VEGF in this process in order to explore the pathogenesis of glomerular disease in preeclampsia. SFlt-1 expression in the human trophoblasts (TEV-1 cells) was enhanced by transfecting sFlt-1 plasmid DNA into TEV-1 cells. The monolayer barrier function of glomerular endothelial cells (ciGEnCs) was determined by measuring the fluorescence intensity of bovine serum albumin (BSA) that crossed the monolayer of glomerular endothelial cells. The results showed that the over-expression of sFlt-1 by TEV-1 cells led to the barrier dysfunction of ciGEnCs, and the exogenous VEGF could alleviate the ciGEnCs dysfunction resulting from the over-expression of sFlt-1 to a certain extent. It was concluded that the dysregulation of sFlt-1 and VEGF in preeclamptic pregnancy may contribute to the barrier dysfunction of glomerular endothelial cells, and VEGF may play an important role in maintaining the barrier function of glomerular endothelial cells, but it may not be the sole factor.

This study investigated the effect of epigenetic modification of maspin on extravillous trophoblastic function. The mRNA expression of maspin in placentae from normotensive and preeclamptic pregnant women was detected by RT-PCR. TEV-1 cells, a human first-trimester extravillous trophoblast cell line, were cultured and treated with CoCl(2) (300 μmol/L) to induce chemical hypoxia and with 5-aza (500 nmol/L) to induce demethylation. The mRNA expression of maspin in TEV-1 cells subjected to different treatments was determined by RT-PCR, and the proliferative and migratory abilities of TEV-1 cells were assessed by cell counting kit-8 (CCK-8) and Transwell assays. Our results showed that the maspin mRNA expression level in placentae from preeclamptic women was much higher than that from normotensive women. CoCl(2) or 5-aza could up-regulate the mRNA expression of maspin and significantly suppress the proliferation and migration of TEV-1 cells. It was concluded that the epigenetic modification in promoter region of maspin contributes to incomplete trophoblast invasion, which offers a novel approach for predicting and treating placental dysfunction.

OBJECTIVETo analyze the causes of misdiagnosis of primary hypothyroidism (PH), with an attempt to reduce the misdiagnosis or mistreatment.

METHODSTotally 70 PH children with a history of misdiagnosis but whose conditions were confirmed in Peking Union Medical College Hospital and the First Hospital of Jilin University from July 2000 to May 2009 were enrolled in this study. The clinical data were collected and the causes of misdiagnoses were analyzed.

RESULTSOf these 70 patients, 19 were misdiagnosed as anaemia and dystrophy, 18 as pituitary tumors, 10 as adiposities, 6 as myocarditis or pericardial effusion, 4 as Downs syndrome, 3 as hepatitis, 3 as amyasthenia, 3 as cerebral palsy, 2 as cystis thyrolingualis, and 2 as congenital megacolon. The duration of misdiagnoses ranged from 6 to 72 months. The clinical manifestations of these patients were complicated, involving multiple organs and systems.

CONCLUSIONSPH has complicated clinical manifestations and individual variations, and therefore can be easily misdiagnosed. Good knowledge, sufficient history-taking, and cautious physical examinations can help avoid misdiagnosis. Neonatal screening is helpful for diagnosis and treatment of congenital hypothyroidism.

Adolescent ; Child ; Child, Preschool ; Diagnostic Errors ; Female ; Humans ; Hypothyroidism ; diagnosis ; Infant ; Male