1.Effect of mulberry leaves extracts on glucose uptake of insulin-resistant HepG2 cells and the mechanism.
Fei FANG ; Mingli LUO ; Nan SU ; Xinrong WU
Acta Pharmaceutica Sinica 2012;47(11):1452-6
The effect and mechanism of mulberry leaves extracts (MLE) on glucose uptake of insulin-resistant HepG2 cells in vitro was explored. The insulin resistant models of HepG2 were induced by high concentration of insulin for 24 h. The models were incubated in a buffer containing mulberry leaves extracts. The glucose consumption was detected by glucose assay kits and the AMP-activated protein kinase (AMPK), Akt activation was examined by Western blotting. Mulberry leaves polysaccharides, mulberry leaves flavonoids and mulberry leaves extracts advanced glucose uptake of insulin-resistant HepG2 cells; Mulberry leaves extracts enhance phosphorylation of AMPK. Mulberry leaves extracts do not change the phosphorylation status of Akt. The glucose consumptions of insulin resistant model of HepG2 were promoted by mulberry leaves extracts. MLE stimulates HepG2 cell AMPK activity acutely without changing the Akt activity.
2.Establishment and evaluation of detection method of bifidobacteria in human fecal using real-time fluorescence quantitative PCR
Liwen ZHANG ; Zongrun WANG ; Xiuli WU ; Mingli FANG ; Yunfeng ZHANG
Journal of Jilin University(Medicine Edition) 2014;(3):686-691
Objective To establish the real-time fluorescence quantitative PCR method for the detection of bifidobacteria in human fecal samples, and to provide an effective means for measuring intestinal bacteria. Methods Total DNA of bacteria was extracted from 60 cases of children's fecal samples. Three primers of bifidobacteria based on the 16S ribosomal RNA (16SrRNA)which possessed specialities of bacteria as amplified region were designed.The part of amplified 16SrRNA gene sequences was used as standard production.The serial dilution of standard was analyzed to build an absolute quantitative standard curve with SYBR GreenⅠ dye method, and the bifidobacterium contents in sixty human fecal samples were calculated. The sensitivity of the reaction was calculated by detecting the lowest detectable standard which determined the sensitivity of the reaction. The PCR products’melting curve was used to evaluate the specificity.The coefficient of variation (CV)of different batches of standard with the same concentration was used to evaluate the stability of reaction.Results The length of PCR product fragment which was used to build the standard curve was about 6 1 3 bp, the sequencing result was consist with the goals, and the standard sample of bifidobacteria was successfully established in real-time fluorescence quantitative PCR.The standard curve showed a good linear relationship with R2=0.999.The minimum detection value was 1.48×102 copies per reaction.The melting curve of real-time fluorescence quantitative PCR was a single peak.The test samples were batched and then examined by fluorescence quantitative PCR.The CV of standards’ Ct values which calculated from the standard (1.48 × 103 -1.48 × 107 copies · μL-1 )were 2.94%, 3.39%, 3.54%,3.08%,and 3.34%,respectively.The contents of bifidobacteria in fecal from 60 children was 7.77± 0.86(copies · g-1 wet fecal)transformed by logarithmic.Conclusion The established real-time fluorescence quantitative PCR method has high sensitivity, strong specificity and good repeatability, which is suitable for detection of human fecal bifidobacteria content.
3.Analysis on association between polymorphism of CD14 and IL-8 gene and susceptibility of necrotizing enterocolitis
Jiayi TIAN ; Tong ZHU ; Jian WANG ; Mingli FANG ; Chaoying YAN
Journal of Jilin University(Medicine Edition) 2016;42(5):958-962
Objective:To investigate the relationship between the gene polymorphism cluster of differentiation 14 (CD14)-159C/T (rs2569190),and interleukin-8 (IL-8)-251A/ T (rs4073)and the susceptibility of necrotizing enterocolitis (NEC),to clarify the influencing factors of susceptibility of NEC and to provide genetics theory basis for the research on the pathogenesis of NEC. Methods:Total 28 newborns with NEC and 41 newborns without NEC were selected.The amplification of peripheral blood DNA was conducted by PCR.The genotypic and allelic frequencies of CD14-159C/T and IL-8-251A/T of the patients were detected by Sanger DNA sequencing method. The relationship between them and the susceptibility of NEC was studied.Results:The distribution of genotypic frequencies of CD14-159C/T and IL-8-251A/T was consistent with Hardy-Weinberg equilibrium (P >0.05).There were no significant differences of the allelic and genotypic frequencies of CD14-159C/T,or genotypic frequencies of IL-8-251A/T between two groups (P >0.05).While in NEC group,the T allelic frequency of IL-8-251A/T site was higher than that in control group (χ2 = 4.184, P = 0.041, OR = 2.14, 95% CI: 1.03 - 4.46 ). Conclusion:The polymorphism of CD14-159C/T is irrelevant to the pathogeny of NEC,but T allelic frequency of IL-8-251A/T site might be related to the susceptibility of NEC.So T allele in IL-8-251A/T may be one of the danger factors of NEC.
4.Practice and reflection on the international remote pathology consultation
Xiangdong YOU ; Jianan WANG ; Wei LI ; Meijuan LAN ; Mingli YANG ; Xu FANG ; Guofeng WANG ; Fangmin GE
Chinese Journal of Hospital Administration 2011;27(8):613-615
Justified the necessity of remote pathology consultation in China, and described the basic approach of such consultation in terms of the conditions, organizational framework, specialists and consultation process of the hospital. on the basis of benefit in pationts, the consultation helps the development of the pathology department and other specialist departments at the same time, builds initially a pathology quality control system, and accelerates the multi-discipline diagnosis and treatment approach. Expect to encourage contemplation on international remote pathology consultation in an effort to improve such a practice for the benefit of patients.
5.Relationship between the CYP2C19 polymorphism and the efficacy of triple therapy with lansoprazole onHelicobacter pylori infection in elderly patients
Junli XU ; Yujie FANG ; Yongguo LIU ; Mingli ZHANG ; Songdi WU ; Yali CUI
The Journal of Practical Medicine 2014;(20):3242-3244
Objective To investigate the Relationship between the CYP2C19 polymorphism and the efficacy of triple therapy with lansoprazole on Helicobacter pylori infection. Methods 125 elderly patients diagnosed with H.pylori infection were treated with triple therapy. Polymorphism of CYP2C19 was measured by AS-PCR. 105 young patients were selected as control group. The relationship between the polymorphism and eradication rate were analyzed. Results Among the 125 patients,eradication rate of extensive metabolizer group,internal metabolizer group and poor metabolizer groups was 85.29%,76%and 89.39%, respectively. There was no significant difference between groups (P > 0.05). Eradication rate showed no difference between experimental and control group either (P>0.05)(P>0.05). Conclusion The results suggests that the CYP2C19 polymorphism has no correlation with the eradication rate of Helicobacter pylori infection by triple therapy with lansoprazole in elderly patients.
6.Relationship between endothelial progenitor cells and cardiovascular diseases in maintenance hemodialysis patients
Yaping ZHAN ; Huili DAI ; Weiming ZHANG ; Mingli ZHU ; Yan FANG ; Renhua LU ; Zhaohui NI ; Jiaqi QIAN
Chinese Journal of Nephrology 2016;32(12):881-887
Objective To investigate the relationship between the variation of endothelial progenitor cells (EPC) number and cardiovascular diseases (CVD) in maintenance hemodialysis (MHD) patients ,and discuss the function of EPC in the progression of CVD in MHD. Methods One hundred and fifteen MHD patients over 18 years whose dialysis vintage was over six months from Department of Nephrology, Renji Hospital, Shanghai Jiao Tong University School of Medicine were enrolled. They were divided into CVD group and non ? CVD group by medical history, electrokardiographie (EKG), cardiac ultrasound, peripheral vascular imaging and cardiovascular imaging. Peripheral blood (5 ml) was collected for detecting EPC number by flow cytometry as CD34/CD133/vascular endothelial growth factor receptor 2 (VEGFR2) cells. The EPC number between CVD group and non?CVD group was compared. The relationship between the decrease of EPC number and CVD risks in MHD patients was analyzed by logistic regression analysis. In a three?year follow?up, the death and new CVD events of the two groups were compared in order to discuss the relationship between EPC number and adverse events. Results Among 115 MHD patients, the average age was 61.57 ± 12.76, male/female was 71/44, the average dialysis vintage was (86.24 ± 56.31) months, the average Kt/V was 1.69 ± 0.29 and average ultrafiltration volume was (2.48 ± 0.90) L. Forty?four patients in 115 (38.3%) were with concurrent CVD. The EPC number in CVD group was significantly lower than that in non CVD group (P=0.015). The CVD group had higher serum phosphate (P=0.013), higher glycosylated hemoglobin (P<0.001), but serum calcium, intact parathyroid hormone (iPTH) and other indicators had no significant difference between two groups. Multiple Logistic regression analysis showed that older age (OR=1.061), history of diabetes (OR=9.796), dialysis vintage (OR=1.015), serum phosphate (OR=3.766), decrease of EPC number (OR=0.909) were the independent impact factors of CVD events in MHD patients. There were 22 patients of the 115 MHD patients had encountered a new CVD event in a three?year follow?up between December 2012 and December 2015, 9 patients from the CVD group and 13 patients from the Non?CVD group, and there was no significant difference between two groups (P=0.776). Nine patients from the CVD group and 7 patients from the Non?CVD group died in the follow?up, and there was no significant difference (P=0.111). Seventy?one MHD patients from the non?CVD group were divided into two groups by the median of EPC number. There were 3 patients in the higher EPC number group encountered CVD events and 10 patients in the lower EPC number group encountered CVD events, which had significant difference (P=0.024). Conclusion The decrease of circulating EPC number may be related with CVD events in MHD patients. Even adjusted by age, sex, diabetes, dialysis vintage and serum phosphate, decreased EPC number is still the independent risk factor of CVD events in MHD patients. The decrease of EPC number in MHD patients may be used to predict the occurrence of cardiovascular events.
7.Practice on the early training of basic scientific research skills for seven-year medical program students in preclinical medical teaching stage
Li YU ; Haihong FANG ; Shujuan LV ; Lingzhi CHEN ; Xiaojuan DING ; Mingli WANG
Chinese Journal of Medical Education Research 2003;0(02):-
The training of basic scientific research skills is very important for seven-year medical program students. Through various kinds of scientific research training in preclinical medical teaching stage, the students’ abilities of scientific thinking, performing experiments, and writing scientific papers have been improved.
8.The effect of Shuanghuanglian on the load of hepatic HCMV DNA in mice infected with HCMV
Jianjun WU ; Guiyu LIU ; Guihai CHEN ; Shu HOU ; Caohui FANG ; Mingli WANG
Chinese Pharmacological Bulletin 1986;0(05):-
Aim To investigate the effect of Shuanghuanglian on the the load of hepatic HCMV DNA in mice infected with HCMV.Method The model of mice hepatitis infected with HCMV was prepared. 10 days after treated with Shuanghuanlian,the hepatic HCMV DNA was detected qualitatively by polymerase chain reaction;the level of alanine aminotransferase (ALT) and aspartate aminotransferase(AST) in serum were assayed by spectrophotometer;and hepatic pathology was observed with HE staining.Results HCMV DNA load in infected model group was 3875.4760 copies/mg,while in high and middling dosage shuanghuanglian therapy group it was 8.3261 copies/mg、9.1476 copies/mg respectively;It decreased significantly compared with the former(P0.05);low dosage of shuanghuanglian(HCMV DNA load was 3812.8980 copies/mg) had no notable difference compared with infected model group;serum transaminase (ALT,AST) levels significantly elevated in infected model group; serum transaminase level decreased sharply and liver histological pathology was lighten after treated with high and middling dosage of Shuanghuanglian(P
9.The expression and clinical significance of circulating tumor cells in small-cell lung cancer
Cheng QIAN ; Mingli GU ; Chaoping FANG ; Lei SHI ; Hu LI ; Yan CHEN ; Chuanlu REN ; Qin QIN ; Anmei DENG
Chinese Journal of Laboratory Medicine 2014;(5):362-364
Objective To investigate the expression of Circulating tumor cells ( CTCs ) in peripheral blood cells of patients with different stages of Small-Cell Lung Cancer ( SCLC) ,and to evaluate its significance in early diagnosis of lung cancer metastasis.Methods Thirty-five patients with SCLC ( 12 in limited stage and 23 in extensive stage ) and 25 patients with benign lung disease were recruited at the Shanghai Changhai Hospital from April 2011 to April 2013.Samples were prepared from 7.5 ml peripheral venous blood and collected in CellSave tubes.The CTC counts were determined using CellTracks AutoPrep fluorescence scanning system according to the manufacturers′instructions.The expression of CTCs in peripheral blood of SCLC patients and benign lung disease patients were analyzed.The expression of CTCs in different stages of SCLC was measured and compared.Furthermore, samples were prepared from 2 ml peripheral venous blood by centrifugation.The serum levels of NSE Neuron specific enolase were measured.The relationship between the expression of CTC and NSE was analyzed.Results CTCs positive rates in SCLC were significantly higher than in benign lung disease controls [ Positive rates of CTC≥1 were 80.0%and 12.0%(χ2 =27.003,P<0.000 1),CTC≥5 were 51.4%and 0 (χ2 =18.367,P<0.000 1), CTC≥10 were 34.3%and 0(χ2 =10.714,P<0.001),CTC≥50 were 17.1 and 0(P=0.036,P<0.05), respectively ].CTCs positive rates in SCLC extensive stage were significantly higher than limited-stage [Positive rates of CTC≥1 were 58.3% and 91.3%(P=0.033,P<0.05), CTC≥5 were 65.2% and 25.0%(P=0.035,P<0.05), respectively].The expression of CTCs was significantly correlated with NSE (r=0.743 7, P<0.000 1).Conclusion The expression of CTC in SCLC patients is significantly higher than those in control group , and is closely related to clinical stages , which may provide new clues to early predicting of SCLC metastasis and deterioration.
10.Correlation of Angiogenic Factor AGGF1 Expression With Prognosis of Patients With Colon Cancer
Chinese Journal of Gastroenterology 2018;23(6):347-350
Background:Angiogenesis plays a critical role in tumor growth and metastasis. AGGF1,a new member of angiogenic factors,has been shown to be aberrantly expressed in a variety of malignant tumors. Aims:To investigate the correlation of AGGF1 expression with prognosis of patients with colon cancer. Methods:Eighty colon cancer patients undergoing surgical operation from Jan. 2010 to Dec. 2012 at Xi’an First Hospital were recruited. Immunohistochemistry was performed to evaluate the expression of AGGF1 in cancerous and paired paracancerous tissues. The correlations of AGGF1 expression in cancerous tissue with clinicopathological characteristics and prognosis were analyzed. Cox regression model was used to identify the prognostic factors of colon cancer. Results:AGGF1 expression was significantly higher in colon cancer tissue than in paracancerous tissue (67. 5% vs. 32. 5%,P<0. 05),and was correlated with tumor differentiation,lymph node metastasis,vascular/lymphatic infiltration and TNM stage (P<0. 05). Kaplan-Meier survival analysis showed that high AGGF1 expression was correlated with decrease of survival rate in colon cancer patients (P <0. 05). Univariate and multivariate analyses identified AGGF1 as a prognostic factor of patients with colon cancer (OR=2. 09,95% CI:1. 67-4. 45,P=0. 011;OR =1. 98,95% CI:1. 72-4. 59,P =0. 000 ). Conclusions:Angiogenic factor AGGF1 is up-regulated and correlated with tumor metastasis and unfavorable prognosis in patients with colon cancer. It might be a potential prognostic indicator and deserves further study.