1.Determination of Strychnine and Brucine in Strychnos Nux-vomica Linn by SPME-HPLC
China Pharmacist 2016;19(5):994-996
Objective:To establish a SPME-HPLC method for the determination of strychnine and brucine in Stryehnos nux-vomiea Linn. Methods:A novel capillary microtube monolithic column coupled with HPLC was prepared for the selective solid-phase microextraction of strychnine and brucine in Stryehnos nux-vomiea Linn. A Hypersil ODS column(150 mm × 4. 6 mm, 5 μm)was used and the mobile phase was acetonitrile- 0. 01 mol·L -1 sodium heptanesulfonate mixed with the same quantity of 0. 02 mol · L -1 potassium dihydrogen phosphate(adjusting pH to 2. 8 with 10% phosphoric acid)(21 ∶79). The detection wavelength was 260 nm,the sample size was 20 μl,and the column temperature was 25℃ . Results:The linear range of strychnine was 1. 2-90. 0μg·ml -1(r = 0. 999 8)and that of brucine was 1. 0-75. 0μg·ml -1(r = 0. 999 6). The average recovery was 100. 05%(RSD = 0. 58% ,n = 6)for strychnine and 99. 98%(RSD = 0. 27% ,n = 6)for brucine. Conclusion:The method is accurate and highly efficient,which can be used for the determination of strychnine and brucine in Stryehnos nux-vomiea Linn.
2. The sphere formation and tumorigenesis of lung cancer stem cells
Tumor 2008;28(9):751-754
Objective: To induce the sphere formation from human lung adenocarcinoma cell line SPC-A1, identify cancer stem cells among the spheres, and evaluate their tumorigenic potential. Methods: The spheres were induced by incubation of cancer cells in the serum-free medium. The sphere formation was induced by epidermal growth factor (EGF), insulin-like growth factor-1 (IGF-1), and fibroblast growth factor-10 (FGF-10). The stem cell-related markers were measured by RT-PCR and immunofluorescent staining in the resultant sphere-forming cells. Their tumorigenesis was tested in NOD/SCID mice. Results: The float-growing spheres (named pulmospheres) were developed after SPC-A1 cells were cultured for 5-10 days. RT-PCR demonstrated that the pulmospheres expressed the lung cancer stem cell surface markers (CD24 and CD221), the bronchioalveolar stem cell markers [surfactant proteins C (SP-C) and Clara cell secretory protein (CCSP)], the embryonic stem cell stemness genes (OCT4, Nanog, and Bmi-1), and the lung stem cell stemness gene (TTF-1). Among these markers, the expressions of SP-C, CCSP, and OCT4 were further evaluated with the immunofluorescent staining. Approximately 80 % of the sphere cells were shown to be positive for the proteins. The pulmosphere cells were highly tumorigenic in NOD-SCID mice, with tumor development after implantation of 5 × 103 cells per mouse. Conclusion: The lung cancer stem cells are enriched in the pulmosphere-forming cell population, which are tumorigenic.
3.The value of the Montreal Cognitive Assessment in diagnosing mild cognitive impairment
Jie XIANG ; Deqin GENG ; Zhaohui QIN
Chinese Journal of Geriatrics 2009;28(11):905-907
Objective To explore the value of Montreal Cognitive Assessment (MoCA) in the diagnosis of mild cognitive impairment. Methods Mini-mental state examination (MMSE) was used in 532 elderly persons aged 60 years and over in Xuzhou city Gulou county. The 69 healthy people and patients with mild cognitive impairment were chosen to undergo MoCA. Then the sensitivity and specificity of MoCA were analyzed. Results According to MMSE, there were 19 patients with mild cognitive impairment (27. 5 %), 50 healthy persons (72.5 %). While according to MoCA, there were 58 patients with mild cognitive impairment (84.1%), 11 healthy persons (15.9 %). The consistency of the two scales was not good. And compared with MMSE, the sensitivity of MoCA was 94.7%, and the specificity was 20. 1%. Conclusions In the diagnosis of mild cognitive impairment, MoCA is more sensitive than MMSE.
5.Effects of Telmisartan on the expression of Toll-like receptor 4 in monocytes of ApoE-deficient mice
Xiaoyong GENG ; Mingzhao QIN ; Bingying NIU
Chinese Journal of Geriatrics 2012;31(1):68-70
ObjectiveTo observe the effects of telmisartan on toll-like receptor (TLR) 4 expression in monocytes of mice atherosclerosis (AS)model, and explore the mechanism of inflammation.MethodsAS mice model were established in ApoE-deficient mice fed with high-fat diet,and treated with telmisartan as intervention group. After 8 weeks, serum low density lipoprotein,triacylglycerol and total cholesterol were detected in group fed with high-fat diet and intervention group.TLR 4 expressions in two groups were analyzed by flow cytometry. Results There was no significant difference in levels of blood lipids between the two groups.TLR4 expression in monocytes in telmisartan intervention group was lower than group fed with high-fat diet (P<0.05).ConclusionsTelmisartan may intervene the inflammation during atherosclerosis by downregulating TLR4 expression in monocytes.
6.Nested-PCR and LAMP in detection of B .burgdor f eri in ticks
Lin ZHANG ; Xuexia HOU ; Zhen GENG ; Qin HAO
Chinese Journal of Zoonoses 2014;(12):1192-1195
ABSTRACT:Nested‐PCR and loop‐mediated isothermal amplification (LAMP) were applied to identify the Borrelia burg‐dor f eri (B .burgdor f eri) in ticks in this study .A total of 112 adult ixodes were collected from vegetation in a forest area and farm animals in Xunhua County ,Qinghai Province and Xinbin County ,Liaoning Province .The ticks were examined for the presence of B .burgdorferi by nested‐PCR and LAMP .Results showed that 12 in 51 samples were found positive in Xunhua County (23 .53% ) .While positive rate in Xinbin County was 29 .51% with 18 samples positive in 61 samples .In total of 112 tick samples ,the PCR‐positive rate was 17 .86% with 20 positive samples identified ,whereas 15 positive samples were con‐firmed with positive rate of 13 .39 % by LAMP assay .There was no significant difference between the two assays (Х2 =0 .85 , P>0 .05) .Results suggest that both nested‐PCR and LAMP could be used in identifying B .burgdorferi in ticks .Combina‐tion of these two assays could improve the testing results .This is the first report of B .burgdorferi in ticks in Xunhua and Xinbin counties ,and helps to complete the database of the infection rate of B .burgdor f eri in ticks in the widely‐forested area of China .
7.Advances in AEG-1 mediated drug resistance
Kexin CHEN ; Chen LI ; Yu QIN ; Guodong YAO ; Jingshu GENG
Practical Oncology Journal 2014;(5):447-450
It has been demonstrated that astrocyte elevated gene 1 ( AEG-1 ) can promote tumor initia-tion and progression .Over expression of AEG -1 is correlated with tumor angiogenesis ,metastasis and chemother-apy resistance of tumor cells of different origins .The present article is a review on the mechanism of AEG -1 me-diated drug resistance .Studies have shown that AEG -1 participates in carcinogenesis through Ha -ras,myc,NF-κB and PI3K/Akt signalling pathways .AEG-1 can also promote autophagy through activating AMP Kinase . Other researchers demonstrate that AEG -1 promotes MDR1 protein translation by up-regulating MDR1 mRNA expression ,and thus increases polyribosome .It is testified that AEG-1 can influence drug susceptibility and ex-pression of MDR gene as a RNA binding protein .Multiple functions of AEG -1 in drug resistance in multiple cancers demonstrate that AEG -1 can be used as a novel target for antitumor drugs .
8.Association between single nucleotide polymorphism in Nrf2 gene and acute mountain sickness
Qianwen GENG ; Jihang ZHANG ; Jun QIN ; Lan HUANG ; Jun JIN
Military Medical Sciences 2015;(2):105-107
Objective To explore the correlation of single nucleotide polymorphism ( SNP ) in Nrf2 gene with acute mountain sickness ( AMS) among Han populations in China .Methods As a nested case-control study , 603 Chinese Han young men who had been quickly exposed to 3700 m were adopted and divided into case group and control group ( 369 vs 234,respectively) by Lake Louise Self-assessment Scoring System(LLSS).The Sequenom Mass Array system was used to detect the SNPs of rs10497511 and rs2364722 in Nrf2 gene.Results Alleles of rs10497511 and rs2364722 were respec-tively detected in both case and control groups , which were T-C and A-G.Statistically significant difference was not found in allele frequencies ( P>0.05 ) .Further analysis showed that there was still no significant difference between the codomi -nant, dominant and recessive genotype models (P>0.05).Conclusion rs10497511 and rs2364722 of Nrf2 gene may not be related to susceptibility to AMS in Chinese Han populations .
9.Mechanism of long non-coding RNA MALAT1 in promoting migration of hepatocellular carcinoma cells
Hui HAN ; Haibei LI ; Pengbo CAO ; Geng QIN ; Gangqiao ZHOU
Military Medical Sciences 2017;41(6):409-414
Objective To investigate whether the long non-coding RNA (lncRNA) metastasis associated lung adenocarcinoma transcript 1 (MALAT1) can act as a competitive endogenous RNA (ceRNA) to promote the migration of hepatocellular carcinoma (HCC) cells.Methods Transient transfection of small interfering RNA (siRNA) against MALAT1 was used to knockdown MALAT1 in HepG2 cells.Transwell assays were employed to assess the migration capabilities of HepG2 cells upon MALAT1 knockdown.RNA pull-down assays were performed to validate the direct binding between MALAT1 and miR-126*.Quantitative reverse transcription PCR (qRT-PCR) and Western blotting assays were used to detect the mRNA and protein levels of the miR-126* target genes.The dysregulation and prognostic significance of MALAT1 and miR-126* were analyzed in the public dataset of The Cancer Genome Atlas (TCGA).Results Compared with the control group, MALAT1 knockdown significantly inhibited the migration of HCC cells.MALAT1, with three miR-126* response elements, directly sponged miR-126* in a sequence-specific manner.The mRNA and protein levels of CXCL12, which was the miR-126* target gene, were significantly down-regulated upon MALAT1 knockdown.The TCGA database showed that MALAT1 was significantly up-regulated in HCC and high expression levels of MALAT1 were significantly associated with poor disease-free survival, whereas an opposite pattern of miR-126* was observed.Conclusion This study suggests that MALAT1 directly sponges miR-126* and upregulates the expression of CXCL12, which in turn promotes the migration of HCC cells.
10.Molecular mechanism of the reversion back towards fetal energy metabolism occurring with the development of cardiac hypertrophy and the role of Carvedilol
Qin HU ; Longgui LI ; Zhaohua GENG ; Feng JIN ;
Journal of Third Military Medical University 2003;0(16):-
Objective To study the reversion of the metabolic gene expression pattern of hypertrophic cardiac and role of Carvedilol and to explore the molecular regulatory mechanism of Carvedilol on attenuating the reversion back towards fetal energy metabolism occurring with the development of cardiac hypertrophy. Methods A model of hypertrophy induced by coarctation of abdominal aorta(CAA) in male Wistar rats was employed and changes of parameters such as hemodynamics, ventricular remodeling parameters, free fatty acid in blood serum and cardiac myocyte and expressions of muscle carnitine palmitoyltransferase Ⅰ (M CPT Ⅰ) and medium chain acyl CoA dehydrogenase (MCAD) mRNA were investigated in the experimental animals in operation group(CAA), sham operation group(SH) and Carvedilol intervention group(CAR) at 16 weeks after operation. Results Significant hypertrophy was found in the left ventricle in CAA group(3.41?1.30 vs 2.46?1.31, P