Effects of morphine and pethidine on the expression of P-glycoprotein in mouse brain microvascular endothelial cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effects of morphine and pethidine on P-glycoprotein (P-gp) expression in mouse brain microvascular endothelial cells and investigate the role of nuclear factor-kappaB (NF-kappaB) signaling pathway in morphine-induced up-expression of P-gp.</p><p><b>METHODS</b>The mouse brain microvascular endothelial cell line (b.END3) was subjected to pre-incubation with NF-kappaB inhibitor PDTC (5 micromol/L) for 1 h followed by stimulation with morphine (1 microg/ml) or pethidine (1 microg/ml) for 24 h. The bEnd.3 cells were then collected for Western blotting for P-gp expression.</p><p><b>RESULTS</b>A 24-h morphine stimulation induced an up-expression of P-gp in bEnd.3 cells by almost 200%. Pethidine in similar conditions did not affect P-gp expression in the cells. PDTC, the specific inhibitor of NF-kappaB, inhibited morphine-induced up-expression of P-gp in the cells.</p><p><b>CONCLUSION</b>Morphine can induce up-expression of endogenous P-gp in mouse brain microvascular endothelial cells. NF-kappaB signaling pathway is involved in the morphine-induced up-expression of P-gp.</p>

Medium- and long-chain fatty acid triacylglycerol reduce body fat and serum triglyceride in overweight and hypertriglyceridemic subjects / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate effects of medium- and long-chain fatty acid triacylglycerols (MLCT) on body fat and serum lipid in overweight and hypertriglyceridemic subjects.</p><p><b>METHODS</b>A double-blind, controlled clinical trial was carried out, in which 112 subjects with hypertriglyceridemia were enrolled and divided into two groups, there were 56 subjects in each group. One group was randomized to consume long-chain fatty acid triacylglycerol (LCT), and the other to MLCT. All volunteers were asked to consume 25 - 30 g test oil daily for consecutive 8 weeks. Anthropometric measurements of body weight, body fat weight, waist circumference(WC), hip circumference(HC), WHR (ratio of WC/HC), total fat weight, subcutaneous fat area, visceral fat area, and serum biochemical variables of glucose, total cholesterols(TC), triglycerides(TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C)were measured at the initial and final time of the study.</p><p><b>RESULTS</b>11 subjects were excluded from the study because of various reasons. Of the 101 included cases, there were 50 (male subject 34, 68.0%) and 51 (male subject 33, 64.7%) subjects left in LCT and MLCT group respectively. The proportion of men in MLCT (64.7%, 33/51) was not significantly different (chi(2) = 0.1227, P > 0.05) compared to those in LCT (68.0%, 34/50). The average age of MLCT was (54.2 +/- 12.5) which was not significantly different (t = 0.39, P > 0.05) compared to those in LCT (53.2 +/- 13.0); Body mass index (BMI) of MLCT was (25.9 +/- 3.3) kg/m(2), which was not significantly different (t = 0.08, P > 0.05) compared to those of LCT (25.9 +/- 2.4) kg/m(2). After consumption of test oil for 8 weeks, extent of decrease in BMI, percent of body fat, subcutaneous fat, serum TG and serum LDL-C in overweight subjects of MLCT were (-0.73 +/- 0.61) kg/m(2), (-1.53 +/- 1.32)%, (-16.29 +/- 19.25) cm(2), (-0.57 +/- 0.86) mmol/L and (-0.05 +/- 0.64) mmol/L respectively, those in overweight subjects of LCT were (-0.19 +/- 0.61) kg/m(2), (-0.58 +/- 1.02)%, (4.69 +/- 19.06) cm(2), (0.65 +/- 1.10) mmol/L and (0.38 +/- 0.58) mmol/L respectively, all of them were significantly different (the value of t were -2.70, -2.43, -3.20, -3.81 and -2.09 respectively, all of P value were less than 0.05).</p><p><b>CONCLUSION</b>Consumption of MLCT can reduce body fat weight and serum triacylglycerol and LDL-C in overweight hypertriglyceridemic subjects under an appropriate dietary regime.</p>

Expression, Purification and Characterization of Gly14-Humanin, a Novel Neuroprotective Peptide / 中国生物工程杂志

Humanin (HN, its analogue [Gly14]-Humanin, HNG) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults. But the relative low content of this peptide in its natural sources limits its further characterization. An expression vector pET32a/HNG was corstructed and transformed it into E. coli BL21 trxB (DE3). HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography. Subsequently, the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC. A 23 mg recombinant HNG (rHNG) from 1 L bacterial culture was purified. The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide. The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence. Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

Expression, Purification and Characterization of Gly14-Humanin, a Novel Neuroprotective Peptide / 中国生物工程杂志

Humanin (HN, its analogue [Gly14]-Humanin, HNG) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults. But the relative low content of this peptide in its natural sources limits its further characterization. An expression vector pET32a/HNG was corstructed and transformed it into E. coli BL21 trxB (DE3). HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography. Subsequently, the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC. A 23 mg recombinant HNG (rHNG) from 1 L bacterial culture was purified. The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide. The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence. Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

A Method of Construction Polycistron Tandem Gene of Small Peptide / 中国生物工程杂志

Objective: To construct a polycistron tandem repeated Echistatin (Ecs) gene. Methods: Three Ecs genes with independent initiation and termination codon were ligated tandem through restriction enzyme sites after amplified with 3 pairs of primers using pMD18T-Ecs as template. The polycistron Ecs gene was inserted into pET30a and expressed in E.coli BL21(DE3) with IPTG induction. The expression results were identified by 18% SDS-PAGE and Western blot. Results: The expression of Ecs polycistron was accomplished with 18% expression level of total protein determined by SDS-PAGE and Western blot. Conclusion: The successful expression of Ecs polycistron provided a new method for the preparation of low molecular weight protein.

Expression,Purification and Characterization of [Gly14]-Humanin,a Novel Neuroprotective Peptide / 中国生物工程杂志

Humanin(HN,its analogue [Gly14]-Humanin,HNG)was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults.But the relative low content of this peptide in its natural sources limits its further characterization.An expression vector pET32a/HNG was corstructed and transformed it into E.coli BL21 trxB(DE3).HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography.Subsequently,the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC.A 23 mg recombinant HNG(rHNG)from 1 L bacterial culture was purified.The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide.The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence.Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

Effects of genistein on bone mineralization in ovariectomized rats / 中国应用生理学杂志

<p><b>AIM</b>To investigate the effects of genistein on bone mineralization in ovariectomized rats.</p><p><b>METHODS</b>Forty-seven Wistar rats were randomly allocated into six groups: sham-operated (sham), ovariectomized (ovx), ovariectomized supplied with diethyl stilbestrol (E, 20 microg x kg bw(-1) x d(-1)) or genistein (25, 50, 100 mg x kg bw(-1) x d(-1)). After the rats had been fed for three months, analysis of the bone mineral density, parameters related to mineralization, bone content of Ca, P, Mg, Mn and Zn and serum concentration of parathyroid calcitonin and estrogen was performed.</p><p><b>RESULTS</b>Bone mineral density, bone Ca, P, Zn and Mg content and serum estrogen concentration in ovariectomized rats were significantly decreased, but mean osteoid width increased, mineralization lag time and osteoid maturation period prolonged compared with sham animals. After three months supplementation to ovariectomized rats, bone Ca, P and Mg content increased, mean osteoid width decreased, mineralization lag time and osteoid maturation period shortened compared with ovariectomized animals.</p><p><b>CONCLUSION</b>Genistein promotes bone mineralization by increasing bone Ca, P, Mg and adjusting serum calcitonin to prevent osteoporosis.</p>

Effects of genistein on proliferation and differentiation of osteoblast / 中国应用生理学杂志

<p><b>AIM</b>To study the effects of genistein on proliferation and differentiation of osteoblasts in neonatal rat calvaria cultures.</p><p><b>METHODS</b>Osteoblasts were isolated from neonatal rat calvaria through trypsin and collagenase digestion, and cultured in the presence of different doses of genistein (10(-5) mol/L, 10(-6) mol/L and 10(-7) mol/L). The proliferation and DNA and collagen synthesis of osteoblasts were assayed by MTT method and 3H-TdR and 3H-proline incorporation. The activity of ALP were measured by ALP assay kit.</p><p><b>RESULTS</b>Genistein significantly increased osteoblast 3H-TdR and 3H-proline incorporation and MTT, 10(-6) mol/L genistein increased ALP activity.</p><p><b>CONCLUSION</b>Genistein increased osteoblast DNA and collagen synthesis in neonatal rat calvaria cultures, and promoted osteoblast proliferation and differentiation.</p>

Effects of zinc deficiency on bone mineralization and its mechanism in rats / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the influence of zinc deficiency on bone mineralization.</p><p><b>METHODS</b>Thirty Wistar rats were randomly divided into three groups with ten in each group, i.e., zinc-deficient group (ZD), control group, and pair-fed group. Histomorphological changes of bone mineralization, bone mineral content and bone density, bone contents of zinc, calcium, phosphorus, magnesium, manganese, copper and hydroxyproline, and serum levels of parathyroid hormone, calcitonin and osteocalcin in the rats were measured.</p><p><b>RESULTS</b>The results showed that the mineral deposit rate and bone contents of zinc, phosphorus and hydroxyproline, and serum levels of calcitonin and osteocalcin lowered significantly in ZD group, as compared with those in the control and pair-fed groups, with (3.26 +/- 0.34) micro m/d, (64.54 +/- 2.34) g/kg, (54.4 +/- 9.5) mg/kg, (9.28 +/- 1.62) g/kg, (41.2 +/- 13.5) micro g/L, (82 +/- 30) micro g/L in ZD group; (5.37 +/- 0.53) micro m/d, (69.01 +/- 4.05) g/kg, (117.4 +/- 8.0) mg/kg, (11.31 +/- 1.30) g/kg, (68.3 +/- 14.4) micro g/L, (131 +/- 46) micro g/L in the control group; and (5.45 +/- 0.30) micro m/d, (67.81 +/- 3.56) g/kg, (106.7 +/- 8.4) mg/kg, (10.88 +/- 1.47) g/kg, (63.7 +/- 12.0) micro g/L, (120 +/- 52) micro g/L in the pair-fed group, respectively. While the time for mineralization lag and osteoid maturation obviously prolonged, (1.08 +/- 0.19) d and (7.12 +/- 2.30) d in ZD group, (0.39 +/- 0.06) d and (2.21 +/- 1.12) d in the control group, and (0.40 +/- 0.06) d and (2.12 +/- 0.58) d in the pair-fed group, respectively. In addition, bone mineral content and bone density and serum parathyroid hormone in ZD group decreased significantly and were lower than those in the control group, but not significantly different from those in the pair-fed group. There were no significant difference in femoral contents of calcium, magnesium, manganese and copper between the ZD group and the control and pair-fed groups.</p><p><b>CONCLUSIONS</b>Zinc deficiency could lower the contents of parathyroid hormone and calcitonin in blood circulation affecting bone mineral deposit and causing defect in bone mineralization.</p>