Screening of housekeeping genes in Gelsemium elegans and expression patterns of genes involved in its alkaloid biosynthesis / 生物工程学报

Gelsemium elegans is a traditional Chinese herb of medicinal importance, with indole terpene alkaloids as its main active components. To study the expression of the most suitable housekeeping reference genes in G. elegans, the root bark, stem segments, leaves and inflorescences of four different parts of G. elegans were used as materials in this study. The expression stability of 10 candidate housekeeping reference genes (18S, GAPDH, Actin, TUA, TUB, SAND, EF-1α, UBC, UBQ, and cdc25) was assessed through real-time fluorescence quantitative PCR, GeNorm, NormFinder, BestKeeper, ΔCT, and RefFinder. The results showed that EF-1α was stably expressed in all four parts of G. elegans and was the most suitable housekeeping gene. Based on the coexpression pattern of genome, full-length transcriptome and metabolome, the key candidate targets of 18 related genes (AS, AnPRT, PRAI, IGPS, TSA, TSB, TDC, GES, G8H, 8-HGO, IS, 7-DLS, 7-DLGT, 7-DLH, LAMT, SLS, STR, and SGD) involved in the Gelsemium alkaloid biosynthesis were obtained. The expression of 18 related enzyme genes were analyzed by qRT-PCR using the housekeeping gene EF-1α as a reference. The results showed that these genes' expression and gelsenicine content trends were correlated and were likely to be involved in the biosynthesis of the Gelsemium alkaloid, gelsenicine.

Study on the sleep quality and its influencing factors in rural hypertensive patients in Xuanen country of Hubei Province / 中华疾病控制杂志

Objective To investigate sleep quality and it’s influencing factors of hypertensives among rural area in Hubei Province. Methods The method of stratified sampling by selecting 569 hypertensives in Xuanen county of Hubei was applied to perform the questionnaire survey, including the sociodemographic data, daily life habits and physical health data, as well as pittsburgh sleep quality index(PSQI) and compliance of hypertensive patients scale(CHPS). Results The average score of PSQI in 569 hypertensives was 7.25±3.61, of which 251(44.11%) hypertensives were poor sleep quality. The influencing factors of sleep quality for hypertensives are gender (P=0.006, OR=1.626), the number of other diseases(P=0.001, OR=1.520), regular exercise (P=0.033, OR=0.660) and the compliance of hypertensives (P=0.024, OR=1.707). Conclusions The sleep quality of rural hypertensives in Xuanen county, Hubei Province is poor, which is affected by different factors. Therefore effective measures should be taken to improve the sleep quality of hypertensives.

Effect of niacin on HDL-induced cholesterol efflux and LXRalpha expression in adipocytes of hypercholesterolemic rabbits / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effect of niacin on HDL-induced cholesterol efflux and LXRalpha expression in adipocytes from hypercholesterolemic rabbits.</p><p><b>METHODS</b>Twelve rabbits fed with high-cholesterol diet for 8 weeks were randomly divided into two groups: (1) high cholesterol group (n = 6): maintained high cholesterol diet for 6 weeks; (2) niacin group (n = 6): the same cholesterol diet plus niacin (0.2 g . kg(-1). d(-1)) for 6 weeks. Control group (n = 6) was fed with normal diet for 14 weeks. Subcutaneous adipose was collected for adipocyte culture. RT-PCR was used to evaluate adipocytes LXRalpha mRNA expressions. Cholesterol efflux rate was determined through measuring release of radioactivity from 3H-cholesterol prelabeled cells into medium containing HDL. The direct effect of niacin on LXRalpha and PPARgamma mRNA expression in primary rabbit adipocytes was assayed.</p><p><b>RESULTS</b>High cholesterol diet resulted in decreased LXRalpha mRNA expressions and reduced cholesterol efflux rate in adipocytes. Six weeks of niacin treatment significantly enhanced the cholesterol efflux from adipocytes, which was related to the increased mRNA expressions of LXRalpha (r = 0.71, P < 0.05). In vitro study, niacin dose-dependently stimulated LXRalpha and PPARgamma mRNA expression in cultured adipocytes and there were positive correlations between various doses of niacin-induced cholesterol efflux and LXRalpha and PPARgamma mRNA expression (r = 0.83 P < 0.01, r = 0.76 P < 0.05, respectively).</p><p><b>CONCLUSION</b>LXRalpha and PPARgamma might play an important role in cholesterol efflux from adipocytes. Niacin can up-regulate LXRalpha and PPARgamma mRNA expressions and promote the cholesterol efflux in adipocytes from hypercholesterolemic rabbits.</p>

Effect of niacin on adiponectin levels in the adipocytes secretion in rabbits / 中南大学学报(医学版)

OBJECTIVE@#To explore the effect of niacin on the serum adiponectin concentration in hypercholesterolemia rabbit and the adiponectin concentration secreted by adipocytes in normal rabbits.@*METHODS@#Ten male New Zealand white rabbits fed with high cholesterol diet for 8 weeks were randomly divided into 2 groups: (1) The high cholesterol group maintained a high cholesterol diet for 8 weeks. (2) The same cholesterol diet plus niacin (0.4g/kg*d ) were administrated for 6 weeks in the niacin group. A control group was fed with normal diet for 14 weeks. Subcutaneous adipose from the control group was collected for adipocyte culture. Matured adipocytes were incubated with various concentrations of niacin (0, 0.25, 0.5, 1.0, and 2.0micromol/L). Adiponectin concentrations in the serum and adipocyte culture supernatant were measured by enzyme-linked-immunosorbent assay.@*RESULTS@#Compared with the control group, rabbits in the high cholesterol group showed higher serum levels of total cholesterol, and low density lipoprotein cholesterol (LDL-C), all of which were significantly reduced by niacin treatment (P<0.01),and serum high density lipoprotein-cholesterol (HDL-C) significantly increased (P<0.01). At 8th week, the mean adiponectin concentration of rabbits fed with high cholesterol diet was significantly lower than that of the control group[(1.268+/-0.039)mg/L vs.(1.449+/-0.107)mg/L,P<0.01]. Niacin treatment significantly elevated the serum adiponectin level which was positively related to HDL-C,and negatively related to TC and LDL-C. Cell experiment in vitro indicated that niacin could significantly induce the adiponectin secretion of adipocytes in a dose-dependent manner.@*CONCLUSION@#Niacin can significantly promote the adiponectin secretion of adipocytes, suggesting that niacin probably has an ability of elevating the serum adiponectin level in addition to lipid-lowering effect.

Effects of oxidized low-density lipoprotein on cholesterol efflux in 3T3-L1 cells / 中南大学学报(医学版)

OBJECTIVE@#To explore whether oxidized low-density lipoprotein (ox-LDL) can stimulate the cholesterol efflux in fully differentiated 3T3-L1 cells and the possible mechanism.@*METHODS@#Fully differentiated 3T3-L1 cells were incubated in the medium containing various concentrations of ox-LDL ( 0 to 50 microg/mL) for 8 or 24 hours. 22(R)-Hydroxycholesterol (10 micromol/L) was exposed to preconditioned adipocytes with 25 microg/mL ox-LDL for 24 hours. Reverse transcription polymerase chain reaction (RT-PCR) was used to evaluate ATP binding cassette transporter A1 (ABCA1), scavenger receptor class B type I (SR-BI), and liver X receptor alpha (LXRalpha) mRNA expression. Cholesterol efflux mediated by apolipoprotein A-I (apoA-I) was determined using liquid scintillator.@*RESULTS@#Low levels (12.5-25 microg/mL) of ox-LDL could increase cholesterol efflux via the enhancement of ABCA1 pathway and SR-BI expression, whereas the higher concentration (50 microg/mL) could not. In adipocytes preincubated with 25 microg/mL ox-LDL for 24 hours, 22(R)-hydroxycholesterol could increase ABCA1 and LXRalpha mRNA and apoA-I-mediated cholesterol efflux, but had no effect on the SR-BI mRNA expression.@*CONCLUSION@#Low levels of ox-LDL may enhance the LXRalpha-ABCA1-apoA-I pathway in adipocytes, up-regulate SR-BI mRNA expression, and then increase the cholesterol efflux. This new effect of ox-LDL will not only make contribution to cholesterol homeostasis in adipocytes, but also be potentially atheroprotective.