The identification of the components absorbed in serum of platycosides in total saponins fraction of Platycodonis Radix is great significance, but there are still great challenges. In this study, 8 types of 44 primordial components from Platycodon saponins were firstly identified using the ultra-performance liquid chromatography-linear ion trap electrostatic field orbitrap high resolution mass spectrometry (UPLC-LTQ-Orbitrap-MS) equipped with the software of Compound Discoverer 3.2 and Trace finder 2.1. Then, the platycosides and their deglycosylated metabolites were used as the template molecules to construct the intestinal microbiota mediated method to identify the primary components and the prototypes in serum. As results, 57 components originating from 44 prototypes of platycosides were identified from drug-containing plasma. Those compounds consist of 12 prototypes of platycosides and 45 metabolites, while the prototypes in serum are also deglycosylated metabolites of other platycosides. The results showed that the intestinal microbiota mediated method could be applied to identify the metabolites of platycosides in total saponins fraction of Platycodonis Radix; and reveal the potential existing forms of prototypes of platycosides in plasma; In addition, it could clearly illustrate the one to many and many to one network of the prototypes and metabolites of platycosides. All animal protocols were approved by the Animal Ethics Committee of Jiangxi University of Traditional Chinese Medicine (No.JZLLSC-202100322).

This study investigated the effects of microRNA-135a (miR-135a) targeting of glycogen synthase kinase 3β (GSK3β) on the epithelial–mesenchymal transition (EMT), migration and invasion of bladder cancer (BC) cells by mediating the Wnt/β-catenin signaling pathway. BC and adjacent normal tissues were collected from 165 BC patients. Western blotting and quantitative real-time PCR were used to detect the expression of GSK3β, β-catenin, cyclinD1, E-cadherin, vimentin and miR-135a in BC tissues and cells. Cells were assigned to blank, negative control (NC), miR-135a mimics, miR-135a inhibitors, small interfering RNA (siRNA)-GSK3β or miR-135a inhibitors+siRNA-GSK3β groups. miR-135a, β-catenin, cyclinD1 and vimentin expression increased, while GSK3β and E-cadherin expression decreased in BC tissues compared with adjacent normal tissues. Compared with the blank and NC groups, the expression of miR-135a, β-catenin, cyclinD1 and vimentin was higher, and cell proliferation, migration, invasion and tumor growth were increased in the miR-135a mimics and siRNA-GSK3β groups. These groups showed an opposite trend in GSK3β and E-cadherin expression and cell apoptosis. The miR-135a inhibitors group was inversely correlated with the blank and NC groups. It was concluded that miR-135a accelerates the EMT, invasion and migration of BC cells by activating the Wnt/β-catenin signaling pathway through the downregulation of GSK3β expression.
Apoptosis ; Blotting, Western ; Cadherins ; Cell Proliferation ; Down-Regulation ; Glycogen Synthase Kinases ; Humans ; Negotiating ; Real-Time Polymerase Chain Reaction ; RNA, Small Interfering ; Urinary Bladder Neoplasms ; Urinary Bladder ; Vimentin

Nonalcoholic fatty liver disease (NAFLD) is a major public health issue worldwide. Immunoglobulin G (IgG) N-glycans are associated with risk factors for NAFLD, such as obesity and diabetes. A cross-sectional study involving 500 Han Chinese adults recruited from a community in Beijing was carried out to explore the association between IgG N-glycans and NAFLD. IgG N-glycosylation was significantly associated with NAFLD, with the disease showing a negative correlation with galactosylation (GP14, GP14n, and G2n), positive correlation with fucosylation (FBG2n/G2n), and positive correlation with bisecting N-acetylglucosamine (GlcNAc) [FBG2n/FG2n and FBG2n/(FG2n+FBG2n)], after controlling age, gender, and prevalence of obesity, type 2 diabetes mellitus, hypertension, and hyperlipidemia. In other words, the present study showed a possible association between NAFLD and the loss of galactose and elevations of fucose and bisecting GlcNAc. Aberrant IgG glycosylation might therefore be a potential biomarker for the primary or secondary prevention of NAFLD.
Biomarkers ; blood ; China ; Cross-Sectional Studies ; Diabetes Mellitus, Type 2 ; blood ; complications ; Female ; Glycosylation ; Humans ; Immunoglobulin G ; blood ; Male ; Middle Aged ; Non-alcoholic Fatty Liver Disease ; blood ; etiology ; Obesity ; blood ; complications ; Odds Ratio ; Polysaccharides ; blood ; Risk Factors

Variations in drug metabolism may alter drug efficacy and cause toxicity; better understanding of the mechanisms and risks shall help to practice precision medicine. At the 21International Symposium on Microsomes and Drug Oxidations held in Davis, California, USA, in October 2-6, 2016, a number of speakers reported some new findings and ongoing studies on the regulation mechanisms behind variable drug metabolism and toxicity, and discussed potential implications to personalized medications. A considerably insightful overview was provided on genetic and epigenetic regulation of gene expression involved in drug absorption, distribution, metabolism, and excretion (ADME) and drug response. Altered drug metabolism and disposition as well as molecular mechanisms among diseased and special populations were presented. In addition, the roles of gut microbiota in drug metabolism and toxicology as well as long non-coding RNAs in liver functions and diseases were discussed. These findings may offer new insights into improved understanding of ADME regulatory mechanisms and advance drug metabolism research.

AIM:To observe the inhibitory effect of siRNA targeting to Wip1 gene on the Wip1 gene expression in the colon cancer cells and to investigate the influence of Wip1 gene silencing on the chemotherapy sensitivity of colon cancer cells.METHODS:Wip1-811 siRNA targeting to Wip1 gene was transfected into RKO colon cancer cells with high expression of Wip1 gene.The mRNA expression of Wip1 was measured by real-time PCR.The protein level of Wip1 was detected by Western blotting.The viability of RKO colon cancer cells was measured by MTS assay.The cell apoptosis and cell cycle were analyzed by flow cytometry.RESULTS: Wip1-811 siRNA efficiently inhibited the expression of Wip1 at mRNA and protein levels.The enhanced chemotherapy sensitivity of RKO colon cancer cells was observed after inhibition of Wip1 gene expression.The viability of RKO colon cancer cells was decreased from (89.4 ±6.6)%to (74.7 ±3.9)%af-ter treated with 5-fluorouracil (P<0.05) and decreased from (77.9 ±2.4)%to (66.7 ±2.9)%after treated with oxali-platin ( P<0.05 ) .The cell apoptotic rate was increased from ( 7.7 ±0.5 )% to ( 12.3 ±3.2 )% and from ( 14.7 ± 2.1)% to (34.0 ±2.1)% when RKO colon cancer cells were treated with 5-fluorouracil and oxaliplatin, respectively (P<0.05).CONCLUSION:Wip1 gene silencing enhances chemotherapy sensitivity of colon cancer cells.

[ ABSTRACT] AIM:To study the process of promoting mouse embryonic stem cells ( ESC) to specify to definitive endoderm by up-regulating of Nodal signal pathway in order to find the best cultivated systems of differentiation of mouse ESC to definitive endoderm cells.METHODS:The cells were divided into different groups based on the culture medium:ESC group ( serum-free medium +LIF) , natural differentiation group ( serum-free medium) and activin A group ( serum-free medium +50μg/L activin A).The cells and the sterilized coverslips with cells were collected at 1, 3, 5 and 7 d of the cultivation.The proportion of CXCR4 +cells was detected by flow cytometry.The expression of CXCR4 was determined by immunocytochemical method, and the protein expression of OCT4 and CXCR4 was detected by Western blot.RE-SULTS:The proportion of CXCR4 +cells showed no dramatic change in ESC group along with the extending of cultivation day, while there were gradually increased in natural differentiation group and activin A group and the highest level was ob-served at 5 d.Among the 3 groups, the proportion of CXCR4 +cells at 5 d was the highest in activin A group.The brown or tan staining in the cells observed under microscope was considered as positive CXCR4 by immunocytochemistry.The pro-tein levels of OCT4 and CXCR4 in ESC group along with the extending of cultivation days was observed.The expression levels of OCT4 were gradually decreased in the cells in natural differentiation group and activin A group, while those of CX-CR4 were gradually increased with the highest level at 5 d.It was highest in the cells in activin A group.CONCLUSION:The proportion of definitive endoderm was the highest at 5 d of the induction during in vitro mouse ESC differentiation.Up-regulation of Nodal signal pathway by adding activin A at the early stage of mouse ESC differentiation promotes mouse ESC to specify to definitive endoderm with CXCR4 molecular marker.

Objective To explore the histopathological features of 850 patients with esophageal malignant tumor in 10 years.Methods From January 2002 to January 2012, 850 patients diagnosed with esophageal malignant tumor were enrolled.Tumor location, general type, pathological type and TNM stage were retrospectively analyzed.All the data were described as case number and percentage.Results Among the 850 cases of esophageal malignant tumor, 33 lesions (3.9％) located in the neck segment of esophagus, 119 lesions (14.0％) located in the upper segment, 44 lesions (5.2 ％) located in the upper-middle segment, 409 lesions (48.1％) located in the middle segment, 123 lesions (14.5 ％) located in the middle-lower segment, 122 lesions (14.4％) located in the lower segment.Among the 724 eases clearly diagnosed as esophageal malignant tumor by general type, the most cases were ulcer type (305 cases, 42.1％), followed by medulla type (260 cases, 35.9％), fungating type (80 cases, 11.0％) and constrictive type (70 cases, 9.7％), and the least cases were intraluminal type (nine cases, 1.2％).Among the 850 cases of esophageal malignant tumor, squamous cell carcinoma (794 cases, 93.4 ％) was the most common cytological type, followed by small cell carcinoma (19 eases, 2.2％), and the least common cytological type was adenocarcinoma (seven cases, 0.8 ％).Among the 724 cases with clear TNM staging, case number of Tis, T1, T2, T3 and T4 stage was eight (1.1％), six (0.8％), 271 (37.4％), 278 (38.4％) and 161 (22.2％), respectively.Among the 122 cases of distal esophageal carcinomas (104 cases with clear TNM staging), most cases were squamous cell carcinoma (112 cases, 91.8 ％), the others cases were adenocarcinoma (three cases, 2.5 ％), small cell carcinoma (three cases, 2.5 ％), basaloid squamous cell, adenosquamous, neuroendocrine carcinomas and carcinosarcoma (one case in each type, 0.8％).Conclusions Esophageal carcinoma was mostly located in the middle segment of in which squamous cell carcinoma was predominant while adenocarcinoma was less common.Esophageal cancer located at lower segment of esophagus is with a wide range of pathological spectrum, squamous cell carcinoma was still dominant, however, esophageal adenocarcinoma is rare.

Objective To explore the relation between clinical-pathological features,Siewert classification and prognosis of esophagogastric junction (EGJ) carcinoma,and to assess the applicability of the new edition of American Joint Committee of Cancer (AJCC) staging guideline on EGJ adenocarcinoma in China.Methods From 2002 to 2012,the clinical data,pathological features,treatment and prognosis of 218 patients with EGJ malignant tumor were retrospectively analyzed.The patients were typed according to Siewert classification criteria and each case was staged according to 7th edition of AJCC TNM staging criteria for esophagus adenocarcinoma and gastric cancer.Kaplan-Meier method and Log-rank test were performed for survival analysis.Results According to the Siewert classification,type Ⅰ was rare (nine cases,4.1％),type Ⅱ was the most common type (150 cases,68.8％) and followed by type Ⅲ (59 cases,27.1％).There was no significant difference in survival curve among the three types (P＞0.05).The survival curve was drawn according to 7th edition of AJCC TNM staging criteria for esophagus adenocarcinoma.In T staging,the prognosis of patients at T4b was better than that of patients at T4a,the prognosis of patients at ⅡB was better than that of patients at ⅡA.The survival curve of patients at Ⅲ C obviously crossed with that of patients at Ⅳ,which was not in conformity with clinical results.The survival curve was drawn according to 7th edition of AJCC staging criteria for gastric cancer.In T staging,the survival curve of patients at Tis was overlapped with that of patients at T1a.The survival rate of patients at ⅡB could not be accurately predicted by the overall staging.In general,the survival of patients with EGJ carcinoma was better predicted according to 7th edition of AJCC staging criteria for gastric cancer than 7th edition for esophagus adenocarcinoma.Conclusions Neither 7th edition of AJCC staging criteria for esophagus adenocarcinoma nor for gastric cancer could accurately predict its prognosis.In our country,EGJ malignant tumor was similar to gastric cancer and had specific clinical-pathological features.It is necessary to research and establish EGJ carcinoma staging criteria instead of applying the current staging criteria for esophagus adenocarcinoma or gastric cancer.

Objective To investigate the clinical characteristics and prognosis of patients with systemic lupus erythematosus (SLE) complicated with acute pancreatitis (AP) .Methods From January 1999 to December 2013 ,the clinical data of 16 patients with SLE complicated with AP among the total 2 526 cases of SLE was collected .A retrospective analysis was performed and the clinical data of patients was classified and documented ,which included general information ,past history ,clinical symptoms , laboratory findings ,imaging findings ,treatment and outcome .The rank sum test was performed for analysis of non‐normal distributed measurement data ,and the Fisher′s exact test was used for count data analysis .Results The incidence of SLE complicated with AP was 0 .63% (16/2 526) .Among them ,ten patients were mild acute pancreatitis (MAP) and six patients were severe acute pancreatitis (SAP) .All patients were treated with fasting ,gastrointestinal decompression ,nutritional support ,anti‐acid ,anti‐inflammatory ,glucocorticoid and somatostatin and so on . Six patients were cured , seven patients improved and three patients died (two SLE complicated with SAP ,one SLE complicated with MAP) . Compared with the SLE patients complicated with SAP ,the SLE patients complicated with MAP were more easily to have lupus nephritis(6/6 versus 5/10 ,Fisher′s exact test) ,hematological system injuries (6/6 versus 5/10 ,Fisher′s exact test) ,liver injuries (5/6 versus 0/10 ,Fisher′s exact test) ,more organs involved (mean 7 versus 3 ,Z= -3 .225) and higher SLE disease active indexes (DAI) score (mean 13 .5 versus 6 .5 ,Z= -2 .876);the differences were statistically significant (all P<0 .05) .Compared with the cured and improved SLE patients complicated with AP ,lupus encephalopathy (2/3 versus 1/13 ,Fisher′s exact test) ,more organs involved (mean 7 versus 5 ,Z= -2 .276) and higher SLE DAI score (mean 21 versus 12 ,Z= -2 .195) was more common in dead SLE patients complicated with AP;the differences were statistically significant (all P< 0 .05) .Conclusions SLE patients complicated with SAP are more easily to get lupus nephritis ,hematological system injuries ,liver injuries ,activity of SLE and multiple‐organ systems involved . The prognosis of SLE patients complicated with AP was poor in those with activity of SLE ,multiple‐organ involved and lupus encephalopathy .

AIM:To investigate the role of side population ( SP) cells in multidrug resistance of colon cancer cells and microRNA biomarkers of SP cells in colon cancer cells .METHODS:SP cells in colon cancer cells were sorted by flow cytometry .The cell viability was measured by MTT method .MicroRNA expression profiles were detected by mi-croRNA chip.MicroRNA expression was verified by real-time PCR.RESULTS:The ratios of SP cells in HCT-15, HT-29 and LoVo colon cancer cell lines were 16.75%, 13.02%and 9.52%, respectively.In all 3 colon cancer cell lines, IC50 of the antitumor drugs including 5-fluorouracil , oxaliplatin and adriamycin for the SP cells were significantly higher than those for non-SP cells (P<0.05).MicroRNA profiling showed that miR-5000-3p, miR-5009-3p and miR-552 were all up-regulated in the SP cells of all 3 colon cancer cell lines .This result was verified by real-time PCR.CONCLUSION:miR-5000-3p, miR-5009-3p and miR-552 are all up-regulated in the SP cells of colon cancer cell lines , and may be the poten-tial microRNA biomarkers of SP cells in colon cancer .