Analysis and identification of volatile components of different parts of Clausena lansium in Hainan by HS-SPME-GC-MS / 中草药

Objective A headspace-solid phase microextraction-gas chromatography-mass spectrometry method (HS-SPME-GC-MS) was adopted for analyzing the volatile components of different parts of Clausena lansium in Hainan Province. Methods Five different fibers were investigated and optimized. Other five experimental parameters such as volume of water, extraction temperature, equilibrium time, extraction time, and salt concentration had been evaluated and optimized by means of the orthogonal design with L16(45) table. Finally, the volatile components of C. lansium leaves, pericarps, and seeds in Hainan were analyzed and identified by GC-MS combined with retention index (RI). Results The optimum extraction conditions were as follows: a 50/30 μm DVB/CAR/PDMS fiber, 10 mg sample powders, 2.0 mL water, 0.2 g NaCl, extraction temperature 80 ℃, equilibrium time 30 min, extraction time 60 min. A total of 83 chemical components were identified from leaves, 96 from pericarps, and 106 from seeds, representing the relative contents of 95.24%, 92.15%, and 95.92% of the total composition. The highest contents were sesquiterpenes in all of the parts, but there were obviously different both in components and contents. Conclusion The HS-SPME-GC-MS method is rapid and sensitive, with a small sample size, without any organic solvents. GC-MS combined with RI has improved the accuracy of analysis and identification. The results may provide experimental basis for further exploitation of C. lansium in Hainan. This method can be used to perform enrichment analysis of the components with high-boiling point and micro-components, which can comprehensively and scientifically characterize and evaluate the quality of Chinese materia medica.

Effect of lentivirus-mediated NOB1 gene silencing by RNA interference on proliferation and apoptosis of human colon cancer cells / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of NOB1 gene on proliferation and apoptosis of human colon cancer cell line RKO by RNA interference.</p><p><b>METHODS</b>Small interference RNA(siRNA) targeting NOB1 gene was cloned into lentivirus vector. Then the lentivirus particles expressing NOB1 short harpin RNA(shRNA) were infected into RKO cells. Real-time PCR and Western blot were performed to examine the expression of NOB1 in lentivirus infected cells. The Thermo Scientific Cellomics ArrayScan VTI HCS Reader was used to test the proliferation and colony-formation of RKO cells, and flow cytometry assay was performed to detect cell cycle and apoptosis. Xenograft tumor was established by injection of RKO cells into nude mice, then NOB1-shRNA was injected into the tumor and tumor volume was detected.</p><p><b>RESULTS</b>Compared to negative controls, the expression levels of NOB1 mRNA and protein were both significantly down-regulated, the proliferation and colony-forming capacity of RKO cells were significantly inhibited, and cell apoptosis was increased after 3 days of NOB1-shRNA lentivirus infection(all P<0.05). The tumor volume was significantly smaller in NOB1-shRNA group than that in Scr-shRNA group[(405±102) mm(3) vs.(870±165) mm(3), P<0.05].</p><p><b>CONCLUSION</b>Silencing NOB1 gene by RNA interference may provide an inhibitive effect on human colon cancer development.</p>

Expression of NOB1 and its significance in colorectal cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of NOB1 in colorectal cancer and its relationship with the clinicopathological characteristics.</p><p><b>METHODS</b>The expression of NOB1 was detected immunohistochemically in 60 primary colorectal cancer tissues and the corresponding normal epithelia (3.0 cm away from the cancer margin) and graded according to the staining intensity and the percentage of positively stained tumor cells.</p><p><b>RESULTS</b>NOB1 overexpression was found in 32 of the 60 cases (53.3%). NOB1 overexpression in the adjacent non-neoplastic tissues was found in 10 of the cases (16.7%), a rate significantly lower than that in the cancer tissues (P<0.05). NOB1 expression was not correlated to such tumor characteristics as gender, age, histological differentiation grade, depth of invasion and lymph node metastasis (P>0.05).</p><p><b>CONCLUSIONS</b>NOB1 expression is higher in colorectal cancer than in normal colorectal tissues, suggesting its involvement in the tumorigenesis and progression of colorectal cancer.</p>

Vector-based RNA interference against vascular endothelial growth factor-C inhibits tumor lymphangiogenesis and growth of colorectal cancer in vivo in mice / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>RNA interference (RNAi) technology is emerging as a very potent tool to generate a cellular knockdown of a desired gene. The aim of this study was to explore whether RNAi targeting vascular endothelial growth factor-C (VEGF-C) could inhibit colorectal tumor lymphangiogenesis and tumor growth.</p><p><b>METHODS</b>We used vector-based RNAi to inhibit VEGF-C expression in colon cancer in vitro and in vivo. VEGF-C expression was quantified by real-time polymerase chain reaction and Westen blotting. To establish LoVo cell tumor xenografts in mice, we subcutaneously inoculated 1.0 x 10(6) LoVo cells in nude mice; after injection, tumors were allowed to grow for 4 weeks until the volume reached (75.8+/-55.8) mm(3). The mice were then randomly divided into two groups: (1) the VEGF-C-siRNA group (n=10) received direct injection of "therapeutic" plasmid 50 microg of LoVo-VEGF-C-siRNA into the tumor mass; (2) the control group (n=10) were injected with LoVo-control in 20 microl of sterile 0.9% NaCl solution into the tumor mass. Tumor growth, microlymphatics and microvessels were compared for mice administered either systemic VEGF-C-siRNA or control over 4 weeks.</p><p><b>RESULTS</b>The mRNA and protein expression of VEGF-C in the colon tumor cell line, LoVo, stably transfected with a VEGF-C-siRNA vector, were significantly downregulated 45.3% and 35.3% respectively. In vivo, four weeks after injection, the tumor volume were significantly smaller in VEGF-C-siRNA group than in LoVo-control group ((314.8+/-54.8) mm(3) vs (553.9+/-90.1) mm(3)); the incidences of lymph node metastasis (30%) in VEGF-C-siRNA were significantly inhibited compared with LoVo-control group (70%); in VEGF-C-siRNA group, the number of microlymphatics per microscopic field was (5.3+/-0.7) and the number of microvessels per microscopic field was (24.2+/-6.5) decreased compared with LoVo-control group (12.5+/-6.9) and (42.1+/-7.4) (all P<0.001).</p><p><b>CONCLUSION</b>Inhibition of VEGF-C expression using siRNA-mediated gene silencing vectors reduced lymphangiogenesis and lymph node metastasis and enhanced survival.</p>

Glycemic patterns and excursions in well-controlled patients with type 2 diabetes mellitus / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To evaluate the glycemic excursions in well-controlled (HbA1c<7%) patients with type 2 diabetes mellitus.</p><p><b>METHODS</b>Thirty-two patients with type 2 diabetes whose HbA1c were<7% underwent CGMS (continuous glucose monitoring system).</p><p><b>RESULTS</b>The highest blood glucose value was in 1.7 h after breakfast. The time duration for blood glucose levels of 7.8 mmol/L, 11.1 mmol/L and 13.9 mmol/L were 28%, 13% and 6%, respectively. The area above 7.8 mmol/L in blood glucose curve was significantly correlated with HbA1c levels. Asymptomatic hypoglycemia and continuous hyperglycemia (BS>13.9 mmol/L more than 2 hours) were also found in the study.</p><p><b>CONCLUSION</b>The apparent hyperglycemia exists in well-controlled type 2 diabetic patients, and CGMS is useful in assessment of glycemia.</p>