1. Separation and purification of naphthaquinone components from rhizoma of Arnebia euchroma(Royle) johnst by medium and low pressure preparative chromatography
Chinese Pharmaceutical Journal 2012;47(11):923-926
OBJECTIVE: To establish a new chromatography technique to, separate and purify naphthaquinone components from rhizoma of Arnebia euchroma(Royle). METHODS: Medium and low pressure preparative chromatography and Flash column were used to separate and purify the constituents, and their structures were elucidated by physicochemical properties, IR, MS, 1H-NMR and C-NMR. RESULTS: Six monocases were obtained, which were deoxyshikonin, shikonin, isobutylshikonin, acetylshikonin, β-ace-toxyisovalerylshikonin, and β, β-dimethylacrylshikonin. The purities of the 6 compounds analyzed by HPLC were 99.62%, 99.02%, 99.5%, 99.31%, 99.3% and 99.2%, respectively. CONCLUSION: A medium and low pressure preparative chromatography method has been established for the first time which can accurately and rapidly separate monocases from herbs.
3.Effects of DNA methyhransferase 1 gene silencing on DNA methylation of pancreatic cancer
Min XU ; Youli ZHANG ; Daojian GAO ; Yuqi ZHANG ; Zhaoshen LI ; Jun GAO ; Yiqi DU ; Yanfang GONG ; Hongyu WU ; Fei GAO
Chinese Journal of Pancreatology 2012;12(4):234-237
ObjectiveTo assess the effects of DNA methyhransferase 1 ( DNMT1 ) gene silencing on DNMTs activity and methylated CpG sites of hMLH-1 in pancreatic cancer cell line PaTu8988.Methods DNMT1 siRNA and negative control siRNA was constructed by Ambion Company of United States.Then they were transfected into pancreatic cancer cell line PaTu8988 at the concentrations of 15,30 nmol/L,and the cells without transfection was used as the control group.Real-time PCR and Western blotting were applied to detect the DNMT1 mRNA and protein expression,and DNMTs activity was detected by using DNMTs activity assay kit.Change of methylation of CpG island of hMLH-1 was detected by bisulfite sequencing PCR (BSP).The expression of hMLH-1 mRNA was detected by Real-time PCR.ResultsAt 48 h after transfection,Realtime RT-PCR analysis showed that the levels of DNMT1 mRNA in DNMT1 siRNA group ( 15 nmol/L) and DNMT 1 siRNA group (30 nmol/L) were 0.573 ± 0.026 and 0.143 ± 0.044,which were significantly lower than those in control group 1.020 ±0.217 and negative siRNA 15 nmol/L group 0.900 ±0.475,and negative siRNA 30 nmol/L group 0.938 ± 0.327 (P <0.05 ).Western blotting analysis showed that the level of DNMT1 protein of DNMT1 siRNA group was also lower than those of negative siRNA and control groups.DNMT activity in DNMT1 siRNA15,30 nmol/L groups was 0.364 ± 0.124and 0.250 ± 0.072,which were significantly lower than those in control group 0.931 ± 0.065and negative siRNA group 0.665 ± 0.055 and 0.472 ± 0.040.DNMT activity was positively correlated with DNMT1 mRNA expression ( r =0.69,P < 0.01 ).DNMT1 RNA interference decreased 8 methylated CpG sites of hMLH-1 to 1 site.Concluslons DNMT1siRNA can specifically inhibit the expression of DNMT1 gene of PaTu8988 and DNMT activity,and can decrease methylated CpG sites of hMLH-1 gene.
4.Expressions of PPAR γ protein during the course of pancreatic fibrosis of chronic pancreatitis in Wistar rats
Ling HUANG ; Jun GAO ; Fei JIANG ; Zheng LU ; Xiaohua MAN ; Aifang XU ; Zhaoshen LI
Chinese Journal of Pancreatology 2011;11(6):426-429
ObjectiveTo observe the expressions of PPAR-γ protein during the course of pancreatic fibrosis of chronic pancreatitis (CP) in Wistar rats and its significance.Methods Bibutyltin dichloride ( DBTC,8 mg/kg body weight) in a volume of 200 ml solvent was injected into the tail vein to establish the CP rat's model.Wistar rats were randomly divided into control group and 1,3,5,7,14,42 d group according to weights.Pancreatic tissue underwent routine pathological examination,and collagen accumulation in pancreatic sections was determined by staining for Sirius Red.Pancreatic myeloperoxidase (MPO)activity was determined.Expressions of α-SMA and PPAR-γ proteins were assessed by immunohistochemical method.Results Light microscopy showed signs of acute pancreatitis with interstitial edema and infiltration of neutrophilic granulocytes 7 d after DBTC injection.Acinar cells necrosis,atrophy,lymphocytes and monocytes infiltration,fibrosis within lobule and peri-lobule as well as pancreatic duct changes were found,which was in accord with the course from AP to CP.One days after induction,the activity of MPO,expressions of α-SMA was significantly increased[ (0.78 ±0.71) vs (0.15 ±0.05)U/g,6.67 ±3.14 vs 0,P<0.05],then it did not increase with time of induction.Seven days after induction,collagen level reached the peak [ (45.42 ±15.99)% ],which was significantly higher than that in control group [ (10.87 ± 2.28 )%,P < 0.05 ].Collagen fibers accumulated from periductal to intra-acinar and/or inter-acinar areas.In control rats,the expression of PPAR-γ protein was positive only in vessel walls,and the expression level was 0.17±0.41 and increased with time of induction,then reached the peak of 4.83 ± 2.71 at 42 d.ConclusionsDuring the course of pancreatic fibrosis in rats,the expression of PPAR-γ protein is gradually increased,and plays a limited anti-inflammation and anti-fibrosis role.
5.The effect of α-tocopherol on fibrosis of chronic pancreatitis rat
Fei JIANG ; Xiaohua MAN ; Yanfang GONG ; Jun GAO ; Zhuan LIAO ; Lianghao HU ; Zhaoshen LI
Chinese Journal of Pancreatology 2011;11(6):430-432
Objective To investigate the effect of α-tocopherol on fibrosis of chronic pancreatitis (CP) rat and explore its mechanism.MethodsMale Wistar rats were randomly divided into control group,acute necrotizing pancreatitis (ANP) group,α-tocopherol group.CP was induced by dibutyltindich loride ( 8 mg/kg) infusion into the tail vein.Gastric lavage of α-tocopherol (800 mg/kg body weight,daily) was started 24 hours after dibutyhindich loride infusion for 4 weeks.The rats in ANP and control group received 0.6 ml salad oil gastric lavage.The rats were sacrificed 4 weeks later.Pancreatic tissue was harvested for histological examination and collagen staining,and measurement of the levels of hydroxyproline and malondialdehyde (MDA) of the pancreas were performed.The mRNA expression of transforming growth factor (TGF)-β1 was measured by real time PCR.ResultsAfter gastric lavage for 4 weeks,the pancreatic tissue inflammation,fiber deposition and abnormal structure in rats of α-tocopherol group were greatly reduced.The levels of MDA and hydroxyproline in rats of α-tocopherol group were significantly lower than those in ANP group [ (0.40 ±0.20) vs (1.07 ±0.41) nmol/100mg,(402.49 ±27.62) vs (664.92 ±29.04) μg/g,P<0.05].The expressions of TGF-β1 mRNA in rats of o-tocopherol group were significantly lower than those in ANP group (2.24 ± 0.89 vs 3.35 ± 0.66,P < 0.05 ).Conclusions Tocopherol gamma can improve pancreatic inflammation and fibrosis by reducing the oxidative stress level and down-regulating the expression of TGFβ1mRNA in rats with CP.
6.Risk factors and outcomes of patients with acute renal injury after intra-coronarystent implantation
Fei HE ; Jun ZHANG ; Zhongqiu LU ; Qingling GAO ; Dujuan SHA ; Guofeng FAN ; Ligang PEI
Chinese Journal of Emergency Medicine 2012;21(5):514-518
Objective To investigate the risk factors and outcomes of acute kidney injury (AKI) in patients after intra--coronary stent implantation.Methods A retrospective and case control study was done with data analysis in 325 patients who underwent intra-coronary stent implantation from January 2010 to March 2011.The patients were divided into two groups as per the criteria of AKI identified on the 7th day after implantation of stent.The variables to be studied included:(1) age,gender,hypertension,diabetes,cerebrovascular disease,left ventricular insufficiency,peripheral angiopathy,creatinine,urea nitrogen,estimated glomerular filtration rate,hyperuricemia,proteinuria,emergency operation,hydration,and medication (ACEI/ARB,statins) before operation; (2) dose of contrast media,operation time,hypotension during intra-operative period; and (3) postoperative:hypotension.The variables were analyzed with the process of One-way ANOVA and multivariate Logistical regression analysis.Consequently,the independent risk factors of AKI in patients after intra-coronary stent implantation could be found.Further,the prognosis of AKI patients was analyzed.Results Of the 325 patients,51 (15.7%) developed AKI.Compared the normal group,hospital stay (P < 0.01 ) and in-hospital mortality (P < 0.05) increased significantly in the AKI group.Monofactorial analysis showed that age,pre-operative laboratory and clinical data including left ventricular insufficiency,peripheral angiopathy,creatinine,urea nitrogen,estimated glomerular filtration rate, hyperuricemia, proteinuria, hydration and emergency operation, and intraoperative information such as operation time and hypotension,and postoperative hypotension in AKI patients group were significantly different in comparison with control group ( P < 0.05 ). Multivariate logistic regression analysis revealed that elderly age (OR =0.253),pre-operative proteinuria (OR =5.351 ),preoperative left ventricular insufficiency ( OR =8.704),eGFR ≤ 60 ml/ ( min · 1.73 m2 ) ( OR =6.677 ),prolonged operation time ( OR =1.017),intra-operative hypotension ( OR =25.245 ) were independent risk factors of AKI ( P < 0.05 ).Conclusions AKI is a common complication and associated with increase in mortality after intra-coronary stent implantation.Increase in age,pre-operative proteinuria,pre-operative left ventricular insufficiency,pre-operative low estimated glomerular filtration rate,prolonged operation time,intra-operative hypotension are the independently risk factors associated with AKI.
7.Cyclooxygenase-2 blockade inhibits accumulation and function of myeloid-derived suppressor cells and restores T cell response after traumatic stress.
Ren-jie, LI ; Lin, LIU ; Wei, GAO ; Xian-zhou, SONG ; Xiang-jun, BAI ; Zhan-fei, LI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(2):234-40
Myeloid-derived suppressor cells (MDSCs) play a crucial role in T cell dysfunction, which is related to poor outcome in patients with severe trauma. Cyclooxygenase-2 (Cox-2) contributes to immune disorder in trauma and infection via production of prostaglandin E2. However, the role of Cox-2 in the accumulation and function of MDSCs after traumatic stress has not been fully elucidated. In the present study, we treated murine trauma model with NS398, a selective Cox-2 inhibitor. Then the percentages of CD11b+/Gr-1+ cells, proliferation and apoptosis of CD4+ T cells were determined. Arginase activity and arginase-1 (Arg-1) protein expression of splenic CD11b+/Gr-1+ cells, and delayed-type hypersensitivity (DTH) response were analyzed. The results showed that Cox-2 blockade significantly decreased the percentages of CD11b+/Gr-1+ cells in the spleen and bone marrow 48 and 72 h after traumatic stress. NS398 inhibited arginase activity and down-regulated the Arg-1 expression of splenic CD11b+/Gr-1+ cells. Moreover, NS398 could promote proliferation and inhibit apoptosis of CD4+ T cells. It also restored DTH response of traumatic mice. Taken together, our data revealed that Cox-2 might play a pivotal role in the accumulation and function of MDSC after traumatic stress.
8.Prevalence of fatty liver in a district of Shanghai detected by B-type ultrasonography and serum activity of alanine aminotransferase
Xiao-Bo CAI ; Jian-Gao FAN ; Jun ZHU ; Xin-Jian LI ; Rui LI ; Fei DAI ;
Chinese Journal of General Practitioners 2005;0(10):-
Objective To study the prevalence of fatty liver and its risk factors in adult population of Pudong New District,Shanghai detected by combination of B-type uhrasonographic features and elevated serum activity of alanine aminotransferase (ALT).Methods A cross-sectional survey was performed in 2017 residents aged 16 years over recruited from four neighborhoods of Prdong New District of Shanghai with multi-phase cluster sampling,including interview with questionnaire,physical check-up,anthropometry, measurement of plasma glucose and lipid profile,ALT activity and real-time B-type ultrasnnography.Serum hepatitis B surface antigen (HBsAg) was further detected for those with elevated ALT activity.Results Prevalence of fatty liver was 21.32 percent (430/2017) in the residents of the District participated in this survey.Prevalence of abdominal obesity,hypertriglyceridemia,hyperlipoproteinemia (low-density lipoprotein-cholecterol),essential hypertension,impaired glucose tolerance,diabetes and metabolic syndrome were 71.16,71.16,11.86,66.74,35.58,24.40 and 47.21 percent in those with fatty liver, respectively,as compared to 26.34,12.73,4.79,39.57,24.01,6.81 and 11.28 percent in those without fatty liver (controls),respectively.Multivariate logistic regression analysis showed that body mass index, 2-h postprandial glucose level,diastolic blood pressure,serum level of triglyceride,abdominal obesity and diabetes all were independent risk factor for tatty liver,with odds ratio (OR) of 1.080,1.149,1.035, 1.526,1.960 and 1.391,respectively.Conclusions Prevalence of fatty liver was relatively high in Shanghai Pudong New District.Fatty liver closely associates with disturbance of carbohydrate and lipid metabolism.
9.Survey of cognitive function in motor neuron disease
Fei GAO ; Dongsheng FAN ; Huali WANG ; Qiong YANG ; Yan ZHANG ; Jun ZHANG ; Yang SHEN
Chinese Journal of Internal Medicine 2009;48(1):31-34
Objective The aim of the study is to investigate the prevalence of cognitive impairment and frontotemporal dysfunction in Chinese patients with motor neuron disease(MND).Methods 100 Datients diagnosed as MND underwent a series of survey including mini mental state examination(MMSE),neuropsychiatry inventory(NPI),Hamilton depression scale(HAMD)and Hamilton anxiety scale (HAMA).Demographics,site of onset,and disease severity-functional rating scale(FRS)were also investigated.Results The prevalence of mild cognitive impairment determined with MMSE score was 24.2%.Compal ison between patients with normal and abnormal MMSE showed statistic differences in depression state and FRS score.Since patients with anxiety and depression would also have abnormal NPI,a follow uP stndy after 3-month antidepressive therapy for the depression patients was made and 2 patients were found to haye Drobably frontotemporal dysfunction lasting for more than 6 months.Conclusion MND patients might have mild impairmented of cognitive function and some of the patients were neuroethologically abnormal.2 of the patients might have frontotcmporal dysfunction.
10.Analysis of oral and maxillofacial malignant tumors implanted with 125I radioactive seeds under guidance of ultrasound
Lang, QIAO ; Jun-fei, GAO ; Ken, WANG ; Zhi-min, ZHANG ; Na, LI ; Kai-ling, XIONG
Chinese Journal of Medical Ultrasound (Electronic Edition) 2010;07(12):2145-2150
Objective To retrospectively analyze 45 cases of oral and maxillofacial malignant tumors and to assess the efficacy and feasibility of oral and maxillofacial malignant tumors implanted with 125I radioactive seeds under guidance of ultrasound.Methods A total of 47 focuses in these 45 patients were determined with the size of these tumors by imaging study,the section was planed by ultrasound,the number and distribution of radioactive seeds were determined with the help of the particle treatment planning system,and were percutaneously implanted particles under guidance of ultrasound.The number and the distribution of particles were assessed by CT.Efficacy endpoints were reexamined and evaluated regularly by ultrasonic and CT according to the standards of WHO.Results The total percentage of efficacy was 70.2% (including complete remission,partial remission).The treatment effect of metastatic carcinoma of lymph node is superior to the parotid tumor.There was no serious complication during the period of implanting and 2 patients with oral ulcers were found after operation.Conclusion The oral and maxillofacial malignant tumor treated implanted with 125I radioactive seeds under guidance of ultrasound is very effective and safe,which is deserved to popularize.The ultrasound is the first choice as a guided method for oral and maxillofacial malignant tumors.