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Objective To screen and identify serum protein biomarkers for the differential diagnosis between ischemic colitis (IC) and ulcerative colitis (UC) by tandem mass tag (TMT) combined with liquid chromatography/tandem mass spectrometry (LC-MS/MS).Methods From January 2018 to January 2019,at the First Affiliated Hospital of School of Medicine of Zhejiang University,patients with UC or IC,and health controls,each l0 cases,were enrolled into UC group,IC group and normal control (NC) group,respectively.Fasting serum samples of all the subjects were collected.After removal of high-abundance protein,followed by proteolysis,peptide labeling and fractionating,the samples were then processed by mass spectrometry.The protein with TMT data of three groups was obtained and protein with TMT value 0 were removed.Heat map of protein was constructed.The differential protein was defined as the protein fold change over 1.5 or less than 0.67.The Reactome database was used to cluster the pathways of differential proteins among groups.Statistical methods included t test,hypergeometry test and corrected by BH multiple test.Results A total of 357 serum proteins were identified by proteomic profiling.There were 27 differential proteins between the IC group and the NC group,including six up-regulated proteins and 21 down-regulated proteins.There were 228 differential proteins between the UC group and the NC group,including 75 up-regulated proteins and 153 down-regulated proteins.There were 49 differential proteins between UC group and IC group,including 22 up-regulated proteins and 27 down-regulated proteins.In the comparison of differential proteins between the NC group,IC group and UC group,only the expression of fibrin 3 was statistically significant (the fold change between UC and NC,between UC and IC,between IC and NC were 0.24,0.46 and 0.53,respectively;t =-5.089,-7.298 and -3.919,all P ＜ 0.01).The results of pathway cluster analysis showed that in the comparison of differential proteins between IC group and NC group,only the platelet degranulation pathway was enriched,and 10 proteins were involved in this pathway (P ＜ 0.01).In the comparison of differential proteins between UC group and NC group,there were 58 pathways enriched,of which 38 proteins were involved in the platelet degranulation pathway,16 proteins were involved in the initial complement trigger pathway,13 proteins were involved in the complement cascade pathway,and 11 proteins were involved in antibody-mediated complement activation pathway (all P ＜ 0.01).In the comparison of differential proteins between UC group and IC group,three different pathways were obtained.Among them,nine proteins were involved in the platelet degranulation pathway,seven proteins were involved in the initial complement trigger pathway,and five proteins were involved in the complement cascade pathway (all P ＜ 0.01).Conclusions The difference in serum proteome between IC patients and UC patients was significant,and the differential proteins are mainly involved in platelet activation and complement activation.The candidate proteins identified in this study may be used as biomarkers for the differential diagnosis of UC and IC in the future.
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Objective@#To screen and identify serum protein biomarkers for the differential diagnosis between ischemic colitis (IC) and ulcerative colitis (UC) by tandem mass tag (TMT) combined with liquid chromatography/tandem mass spectrometry (LC-MS/MS).@*Methods@#From January 2018 to January 2019, at the First Affiliated Hospital of School of Medicine of Zhejiang University, patients with UC or IC, and health controls, each 10 cases, were enrolled into UC group, IC group and normal control (NC) group, respectively. Fasting serum samples of all the subjects were collected. After removal of high-abundance protein, followed by proteolysis, peptide labeling and fractionating, the samples were then processed by mass spectrometry. The protein with TMT data of three groups was obtained and protein with TMT value 0 were removed. Heat map of protein was constructed. The differential protein was defined as the protein fold change over 1.5 or less than 0.67. The Reactome database was used to cluster the pathways of differential proteins among groups. Statistical methods included t test, hypergeometry test and corrected by BH multiple test.@*Results@#A total of 357 serum proteins were identified by proteomic profiling. There were 27 differential proteins between the IC group and the NC group, including six up-regulated proteins and 21 down-regulated proteins. There were 228 differential proteins between the UC group and the NC group, including 75 up-regulated proteins and 153 down-regulated proteins. There were 49 differential proteins between UC group and IC group, including 22 up-regulated proteins and 27 down-regulated proteins. In the comparison of differential proteins between the NC group, IC group and UC group, only the expression of fibrin 3 was statistically significant (the fold change between UC and NC, between UC and IC, between IC and NC were 0.24, 0.46 and 0.53, respectively; t=-5.089, -7.298 and -3.919, all P<0.01). The results of pathway cluster analysis showed that in the comparison of differential proteins between IC group and NC group, only the platelet degranulation pathway was enriched, and 10 proteins were involved in this pathway (P<0.01). In the comparison of differential proteins between UC group and NC group, there were 58 pathways enriched, of which 38 proteins were involved in the platelet degranulation pathway, 16 proteins were involved in the initial complement trigger pathway, 13 proteins were involved in the complement cascade pathway, and 11 proteins were involved in antibody-mediated complement activation pathway (all P<0.01). In the comparison of differential proteins between UC group and IC group, three different pathways were obtained. Among them, nine proteins were involved in the platelet degranulation pathway, seven proteins were involved in the initial complement trigger pathway, and five proteins were involved in the complement cascade pathway (all P<0.01).@*Conclusions@#The difference in serum proteome between IC patients and UC patients was significant, and the differential proteins are mainly involved in platelet activation and complement activation. The candidate proteins identified in this study may be used as biomarkers for the differential diagnosis of UC and IC in the future.
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This study aims to develop a liquid chromatography with tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of ivabradine and N-demethylivabradine in human plasma, and investigate effects of stable isotope labeled (SIL) internal standard (IS) on ivabradine. The analytes and IS were extracted from plasma by protein precipitation with acetonitrile, and chromatographied on a Capcell PAK C18 (100 mm x 4.6 mm, 5 μm) column using a mobile phase of methanol and 5 mmol x L(-1) ammonium acetate. Multiple reaction monitoring with electrospray ionization (ESI) was used in the positive mode for mass spectrometric detection. The effect of ivabradine isotope peak [M+H+3] + on IS and the effect of SIL IS purity on ivabradine were evaluated. An appropriate concentration of SIL IS was chosen to permit method selectivity and linearity of the assay over the required range. The standard curves were demonstrated to be linear in the range of 0.100 to 60.0 ng x mL(-1) for ivabradine, and 0.050 0 to 20.0 ng x mL(-1) for N-demethylivabradine. The intra and inter day precision and accuracy were within the acceptable limits for all concentrations. Besides, the interaction between IS and ivabradine did not impact the determination of analytes. This method was successfully applied to a pharmacokinetic study of hydrogen sulfate ivabradine sustained release tablets on Chinese healthy volunteers.
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Objective To investigate the clinical significance of serum anti-Saccharomyces cerevisias antibody (ASCA),anti-outer membrane porin C (anti-OmpC),antibody to Pseudomonas fluorescens-associated sequence I2 (anti-I2 )and antibody to bacterial flagellin (anti-CBirl )in the diagnosis and treatment of inflammatory bowel disease (IBD).Methods From 2011 to 2013,87 patients with IBD were enrolled and divided into Crohn′s disease (CD)group (66 cases)and ulcerative colitis (UC)group (21 cases).A total of 62 age and gender matched healthy individuals were enrolled as the control group. Fasting blood samples (2 mL)of the subjects were collected.The expression of ASCA,anti-OmpC,anti-I2 and anti-Cbirl antibodies was detected with enzyme-linked immunosorbent assay (ELISA)kits.The diagnosis value of each antibody in IBD and the differential diagnostic value of in UC and CD were compared by receiver operating characteristic (ROC)curve.Results The area under the curve (AUC)of ASCA between IBD and the healthy control group,between CD group and UC group was 0.580 and 0.512, respectively;the accuracy in diagnosis was low.The AUC of anti-CBirl between IBD and the healthy control group was 0.617.There was no differential diagnosis significance of the other antibodies.The positive rate of ASCA in IBD group was 62.1 % (54/87),which was significantly higher than that in the control group (38.7%,24/62).The positive rates of anti-OmpC and anti-I2 in IBD group was significantly lower than those in the control group and the differences were statistically significant (both P 0.05).The specificity,sensitivity,positive predictive value (PPV)and negative predictive value (NPV)of ASCA in differential diagnosis of CD and UC was 52.4%,66.7%,81 .48% and 33.33%,respectively.The specificity and sensitivity of anti-OmpC,anti-I2 and anti-CBirl in differential diagnosis of CD and UC was 81 .0% to 100.0% and 9.1 % to 37.9%,respectively.The specificity,sensitivity,PPV and NPV of double-positive ASCA and anti-I2 in the diagnosis of CD was 57.1 %,86.4%,82.6% and 50.0%, respectively.The positive rate of ASCA and anti-I2 in CD group was significantly higher than that in UC group (84.8%(56/66)vs 57.1 % (12/21 );χ2 =5 .633,P =0.018 ).Conclusions Positive ASCA has some significance in the diagnosis of patients with IBD in our country.The detection of anti-I2 can help to diagnose ASCA negative CD.Because of low sensitivity and positive rate,anti-OmpC and anti-CBirl have limited value in the diagnosis of IBD and the differential diagnosis of UC and CD in our country.
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Objective To assess the impact of intervention with Insulin therapy management unit program in the poorly controlled type 2 diabetic patients in a community of Shanghai.Methods There were 55 patients with type 2 diabetes with poorly controlled hyperglycemia (HbA1C ≥ 8％) enrolled in this study.They were divided at random into 2 groups:intensive care group and standard care group.The subjects in intensive care group were provided with a glucose meter and required to monitor their blood glucose levels at least 2 or 3 times per day.Community health provider acquired information of blood glucose level,episodes of hypoglycemia,and dosage of insulin every week by cell phone.Standard care patients received diabetes care from the same provider in outpatient clinic every month.Results By the end of 6 months of intervention,the intensive care group showed a significant difference in HbA1C as compared to the standard care group (7.40％ ±0.91％ vs 8.65 ％ ± 1.28％,P＜0.01).The frequency of self-monitoring of blood glucose (SMBG) was 4 times per month in both groups at baseline.After intervention,the frequency of SMBG in intensive care group was greatly increased compared to standard care group (50times per month vs 5 times per month,P＜0.01).The frequency of self-reported hypoglycemia in intensive care group was increased compared to standard care group (3 times per month vs once per month,P＜0.01).The average daily dose of insulin in intensive care group was increased 6 units by the end of the present study(P＞0.05).Conclusion After 6 months of intervention,the glycemic control was obviously improved in type 2 diabetic patients treated with insulin and the daily dose of insulin was not increased significantly.TheInsulin therapy management unit is effective and safe.
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<p><b>OBJECTIVE</b>The content of vindoline, catharanthine and vinblastine in the root, stem, leaf, flower and fruit of Catharanthus roseus at various developmental stages were determined, and the biomass allocation was also determined to find the best harvest time.</p><p><b>METHOD</b>The content of vindoline, catharanthine and vinblastine in the root, stem, leaf, flower and fruit of C. roseus were determined by HPLC.</p><p><b>RESULT</b>The content of these alkaloids were influenced by season and it varied in the different tissues of the plant. The content of vindoline and catharanthine in the leaves were the highest, and there was no vindoline detected in the root, but the content of vinblastine in the flower was the highest; the content of vindoline and catharanthine reached the maximum between the August and September, and the content of vinblastine reached the highest after the September. The biomass was the highest in the initial stage of September.</p><p><b>CONCLUSION</b>The best harvest time was in the initial stage of September.</p>
Subject(s)
Catharanthus , Chemistry , Metabolism , China , Chromatography, High Pressure Liquid , Flowers , Chemistry , Metabolism , Plant Extracts , Metabolism , Plant Leaves , Chemistry , Metabolism , Plant Roots , Chemistry , Metabolism , Seasons , Vinblastine , Metabolism , Vinca Alkaloids , MetabolismABSTRACT
An electrochemical L-lactate biosensor was fabricated by combining Platinum nanoparticles (Pt-nano) with multi-walled carbon nanotubes(MWCNTs).L-lactate oxidase(LOD) was immobilized on the surface of the glassy carbon electrode (GCE) modified with MWCNTs and Pt-nano.The surface of resulting LOD/MWCNTs/Pt-nano electrode was covered by a thin layer of sol-gel to avoid the loss of LOD and to improve the anti-interference ability.The cyclic voltammetric results indicated that MWCNTs/Pt-nano catalyst displayed a higher performance than MWCNTs.Under the optimized conditions, i.e., applied potential of 0.5 V, pH 6.4, 25 ℃, the proposed biosensor's determination range was 0.2-2.0 mmol/L, response time was within 5 s, and the sensitivity was 6.36 (A/(mmol/L).It still kept 90% activity after 4 weeks.The fabricated biosensor had practically good selectivity against interferences.The results for whole blood samples analyzed by the present biosensor showed a good agreement with those analyzed by spectrophotometric method.
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To evaluate the effects of San'ao decoction (SAD) and its analogous prescriptions (APs), compounds of traditional Chinese herbal medicine for asthma, on airway inflammation in mice with respiratory syncytial virus (RSV)- and ovalbumin (OVA)-induced asthma.
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Objective To observe the effect of Ruzengping granule on mammary gland of rat with mastoplasia. Methods After benzoate estradiol and progestogen had been injected into rats for 35 days, the mastoplasia model was made. The model rats were treated with Ruzengping granule for 30 days, to measure the mammary papilla diameter of the rats in all groups, to observe morphologic changes of mammary papilla and the changes of mammary gland under microscope, to determine the content of estradiol in blood serum and blood rheology by radio-immunity. Results Ruzengping granule could significantly inhibit the mammary papilla's diameter of the mastoplasia rats, lessen mammary papilla’s symptoms in morphology, reduce the quantity of lobule of mammary gland, lobule-alveolus in mammary gland and the diameter of acinar lumina. Ruzengping granule could also significantly improve blood rheological nature and degrade content of estradiol in blood serum. Conclusion Ruzengping granule can healing efficaciously rat’s mastoplasia induced by benzoate estradiol.
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Aim To study the mechanisms of Pseudolaric acid B(PAB)-induced MCF-7 cell apoptosis and mitotic arrest.Methods MTT assay was performed to assess the cell growth inhibition,contrast phase microscope was used to observe cellular morphologic alteration,and the change of DNA was detected by fluorescent microscopy.The distribution of cell cycle was determined by flow cytometric analysis of propidium iodide staining,and the protein expression was examined by Western blot analysis.Results PAB inhibited MCF-7 cell growth in a dose-and time-dependent manner.4 ?mol?L-1 PAB induced DNA condensation at 24 h.PAB cleaved PARP in a time-dependent manner.At 36 h,PAB up-regulated the expression of cdc 2 and nuclear cyclin B1.Fas antagonistic antibody UB2 had no effect on apoptosis,but agonistic antibody CH11 enhanced the apoptosis induced by PAB.UB2 exerted no effect on cell cycle arrest,and CH11 had the same action as UB2 except for reducing the mitotic arrest through enhancing apoptotic subdiploid peak.Conclusion PAB inhibited MCF-7 cell growth through mitotic arrest and apoptosis.Apoptosis and mitotic arrest were independent of Fas pathway.