Alcohol promotes epithelial mesenchymal transformation-mediated premetastatic niche formation of colorectal cancer by activating interaction between laminin-γ2 and integrin-β1

BACKGROUND Colorectal cancer(CRC) is a common malignant tumor. Alcohol consumption is positively correlated with CRC malignant metastasis;however, the mechanism is unclear. The interaction between laminin-γ2(LAMC2) and integrin-β1(ITGB1) plays a role in premetastatic niche signaling, which may induce epithelial mesenchymal transformation(EMT) and lead to metastasis.AIM To investigate the effects of alcohol on CRC metastasis from the molecular mechanism of the premetastatic niche.METHODS The interaction between LAMC2 and ITGB1 was measured by Duolink assay, and the expression levels of LAMC2, ITGB1 and focal adhesion kinase(FAK), snail, fibronectin, N-cadherin and special AT-rich sequence binding protein 1(SATB1) were measured by quantitative real-time polymerase chain reaction, immunohistochemistry and western blotting. Interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α) and IL-6 levels were measured via enzyme-linked immunosorbent assay, histopathological assessment via hematoxylin eosin staining, and determination of aberrant crypt foci via methylene blue.RESULTS The lymph node metastasis rate was higher in the alcohol group than non-alcohol group. There was a significant increase in interaction signals between LAMC2 and ITGB1, and an increase in phosphorylate-FAK/FAK, snail, fibronectin, N-cadherin and SATB1, whereas E-cadherin was reduced in the alcohol group compared to the non-alcohol group in both animal and clinical samples. Serum IL-1β, TNF-α and IL-6 were higher in alcohol group than in non-alcohol group. Alcohol may promote CRC metastasis by influencing the molecular mechanism of the premetastatic niche.CONCLUSION Our study suggests that alcohol promotes EMT-mediated premetastatic niche formation of CRC by activating the early interaction between LAMC2 and ITGB1 and lead to CRC metastasis.

Effect of Bu-Zhong-Yi-Qi-Tang on deficiency of N-glycan/nitric oxide and islet damage induced by streptozotocin in diabetic rats

AIM: To investigate the effect of Bu-Zhong-Yi-Qi-Tang (Decoction for Reinforcing Middle Jiao and Replenishing Qi) on deficiency of N-glycan/nitric oxide (NO) and islet damage induced by injecting two medium doses of streptozotocin (STZ). METHODS: Diabetes was induced by intraperitoneal injection of STZ at 55 mg/kg on day 1 and day 8. Islet damage was evaluated using a scoring system. Nitrite, nitrate, α-mannosidase and amylase activities were measured by colorimetry. N-glycan patterns of amylase were determined with lectin [ConA, pisum sativum agglutinin (PSA), peanut agglutinin (PNA), and lens culinaris agglutinin (LCA)] affinity precipitation method. RESULTS: Severe islet necrosis and mild islet atrophy were observed in diabetic rats. The number and size ofislets, the activities of α-mannosidase, amylase and nitrite were decreased, while the binding of PNA and LCA to amylase was increased. All of which were improved after treatment with Bu-Zhong-Yi-Qi-Tang. Islet damage was significantly correlated with nitrite, nitrate, α-mannosidase, amylase and the binding of LCA, PNA, and PSA to amylase.CONCLUSION: STZ-induced islet damage is related to N-glycan deficiency in proteins by blocking α-mannosidase activity and no deficiency, accumulation of unfolded proteins, and endoplasmic reticulum stress and activation of cellular signals, all of which are improved after treatment with Bu-Zhong-YiQi-Tang.

Discussing the six meridional diseases of Shanghan Lun based on stress theory

As the theoretical source of Traditional Chinese Medicine clinical medicine,Shanghan Lun plays a vital role in guiding the diagnosis and treatment of clinical diseases.Stress is a systemic nonspecific and adaptive response that occurs when the body is stimulated by internal and external environmental factors.This paper discusses the correlation between the three phases of stress containing alarm,resistance,and exhaustion and the three yin and three yang diseases.It was concluded that sanyang diseases were related to the alarm stage and resistance stage,and sanyin diseases were related to the failure stage.The results indicated the essence of diseases of six meridians from the perspective of stress and provided reference for the diagnosis and treatment of six classics diseases.

Meta-analysis about Western Medicine combined with activating blood drugs on modulating blood glucose and lipids in diabetic patients

OBJECTIVE： To assess the effect of activating blood drugs （AB drugs） on blood glucose and lipid levels of diabetic patients METHODS： Systematic searches were conducted on 6 electronic databases and the search time was cut off in May 2016. Randomized controlled trials （RCTs） were included if they investigated diabetic patients who received Western medicine combined withAB drugs in the experimental groups and solitary usage of corresponding Western medicine or placebo in the control groups. 6 outcome measures, which were considered as inclusion, were changes in fasting blood glucose （FBG）, glycosylated haemoglobin A1 c （HbA 1 c）, total triglyceride （TG）, total cholesterol （TC）, low density lipoprotein cholesterol （LDL-c） and high density lipoprotein cholesterol （HDL-c）. The literature quality was assessed by Cochrane score for risk of bias in Review Manager 5.2, as well as meta-analysis was performed. RESULTS： 7 RCTs comprising 999 patients met all inclusion criteria. Meta-analysis showed beneficial effects of Western medicine combined with AB drugs on modulating FBG, TC, LDL-c and HDL-c compared with Western medicine or placebo （P〈0.05）, while no difference was observed in HbAlc, and TG （P〉0.05）. To some extent, the therapeutic effects of Western medicine combined with AB drugs were better than that of solitary usage of Western medicine or placebo. Besides, AB drugs were safe and reliable with little toxic and side-effects. CONCLUSIONS： The meta-analysis suggests beneficial effects of Western medicine combined with AB drugs on modulating blood glucose and lipids in diabetic patients.

Polysaccharide extracts of Astragalus membranaceus and Atractylodes macrocephala promote intestinal epithelial cell migration by activating the polyamine-mediated K^+ channel

Astragalus membranaceus(Radix Astragali, RA) and Atractylodes macrocephala(Rhizoma Atractylodis Macrocephalae, RAM) are often used to treat gastrointestinal diseases. In the present study, we determined the effects of polysaccharides extracts from these two herbs on IEC-6 cell migration and explored the potential underlying mechanisms. A migration model with IEC-6 cells was induced using a single-edged razor blade along the diameter of cell layers in six-well polystyrene plates. The cells were grown in control--media or media containing spermidine(5 μmol·L^(-1), SPD), alpha-difluoromethylornithine(2.5 mmol·L^(-1), DFMO), 4-Aminopyridine-(40 μ-mol·L^(-1), 4-AP), the polysaccharide extracts of RA or RAM(50, 100, or 200 mg·L^(-1)), DFMO plus SPD, or DFMO plus polysaccharide extracts of RA or RAM for 12 or 24 h. Next, cytosolic free Ca^(2+)([Ca^(2+)]cyt) was measured using laser confocal microscopy, and cellular polyamine content was quantified with HPLC. Kv1.1 mRNA expression was assessed using RT-qPCR and Kv1.1 and RhoA protein expressions were measured with Western blotting analysis. A cell migration assay was carried out using Image-Pro Plus software. In addition, GC-MS was introduced to analyze the monosaccharide composition of both polysaccharide extracts. The resutls showed that treatment with polysaccharide extracts of RA or RAM significantly increased cellular polyamine content, elevated [Ca^(2+)]cyt and accelerated migration of IEC-6 cells, compared with the controls(P < 0.01). Polysaccharide extracts not only reversed the inhibitory effects of DFMO on cellular polyamine content and [Ca^(2+)]cyt, but also restored IEC-6 cell migration to control level(P < 0.01 or < 0.05). Kv1.1 mRNA and protein expressions were increased(P < 0.05) after polysaccharide extract treatment in polyamine-deficient IEC-6 cells and RhoA protein expression was increased. Molar ratios of D-ribose, D-arabinose, L-rhamnose, D-mannose, D-glucose, and D-galactose was 1.0:14.1:0.3:19.9:181.3:6.3 in RA and 1.0:4.3:0.1:5.7:2.8:2.2 in RAM. In conclusion, treatment with RA and RAM polysaccharide extracts stimulated migration of intestinal epithelial cells via a polyamine-Kv1.1 channel activated signaling pathway, which facilitated intestinal injury healing.

Bioinformatics Research on Chronic Superficial Gastritis of Pi-deficiency Syndrome by Gene Arrays

Objective： To determine the bioinformatical characteristics of differential gene expression in patients with chronic superficial gastritis （CSG） with the Pi-deficiency syndrome （PDS） and those of the non- Pi-deficiency syndrome （non-PDS）, i.e. patients of CSG with Pi （脾）-Wei （胃） dampnese-heat syndrome and healthy persons. Methods： With the BRB-Array Tools software package, original data collection and bioinformatic analysis of gene arrays were conducted in 6 CSG patients of PDS （CSG-PDS）, 6 CSG patients of non-PDS （CSG-nPDS）, and 6 healthy volunteers （Normal）. Results： Compared with non-PDS, the gene expressions in PDS with regards to protein synthesis, energy metabolism, immune reaction and ionic transport tended to be down-regulated, while those concerning secretion, cytoskeleton and ubiquitinization were up-regulated dominantly. Conclusions： The two kinds of samples, CSG-PDS/Normal and CSG-PDS/CSG-nPDS, have their respective gene expression profiles with different characteristics. Gene expression profile has certain referential significance in syndrome classification.

Differential expression of immune-related genes between healthy volunteers and type 2 diabetic patients with spleen-deficiency pattern

OBJECTIVE: To investigate the clinical differentiation of spleen-deficiency pattern(SDP), a group of symptoms and signs defined in terms of Traditional Chinese Medicine for its clinical practice.METHODS: Peripheral venous blood(> 3 m L) was collected from each of six type 2 diabetes mellitus(T2DM)-SDP patients and six healthy volunteers. After the isolation of peripheral white blood cells(PWBCs), total RNA was extracted, and quality control was performed on all RNA samples. Microarray experiments were conducted using the Agilent human whole genome gene chip, and genes demonstrating differential expression were screened. Bioinformatics analysis was conducted on these genes using several online databases.RESULTS: We screened a total of 175 differentially expressed genes(DEGs), of which 111(63%) were down-regulated and 64(37%) were up-regulated in T2DM-SDP patients compared with healthy controls. Among the 175 genes, 158 had biological function annotations: 46(29%) were directly related to an individual's immune regulation or response, 25(16%) were associated with substance and energy metabolism of PWBCs which could also indirectly influence immunity, and the remaining87(55%) were involved in a variety of PWBC biological processes that might eventually influence the immune function. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis revealed that the DEGs were predominantly enriched in seven immune-related pathways. Hierarchical cluster analysis identified gene expression patterns that were distinguishable between the two study groups.CONCLUSION: Our results suggest that T2DM-SDP patients experience significant hypoimmunity and/or immune dysfunctions, and possess a specific gene expression profile. These findings offer new insights into SDP and the clinical pattern differentiation of T2DM-SDP.

Study on Clinical Effect and Mechanism of Jianpi Qingre Huayu Recipe (健脾清热化瘀方) in Treating Patients with Gastric Ulcer

Objective： To study the effects of Jianpi Qingre Huayu Recipe （健脾清热化瘀方, JQH） in curing gastric ulcer and to preliminarily probe into its pathogenic mechanism, Methods： Fifty patients with gastric ulcer of Pi （脾）-insufficiency and stasis-heat syndrome type were assigned to the treated group （30 patients） and the control group （20 patients）. They were treated respectively with JQH and Ranitidine. At the same time, another group consisting of 20 healthy persons was set up for normal control. The clinical effect on gastroscopic figure and traditional Chinese medicine （TCM） syndrome were observed. Changes of T-cell subsets and interleukin-8 （IL-8） in serum as well as IL-8 in mucosa around the gastric ulcer were determined before and after treatment by flow cytometry and ELISA. Results： Comparison of the total effective rate on gastroscopic figure in the treated group and the control group （86.7% vs 80.0%） showed insignificant difference, but the cure rate and markedly effective rate in the former （50.0% and 20.0%） was higher than that in the latter （40.0% and 15.0%） respectively. Comparison of the total effective rate on TCM syndrome in the treated group and in the control group （96.7% vs 70.0%） showed insignificant difference, but the cure rate and markedly effective rate in the former （63.3% and 23.3%） was higher than that in the latter （50.0% and 20.0%） respectively. Serum levels of CD3^＋, CD4^＋, CD8^＋ got restored to normal range in the treated group after treatment but it was not so in the control group. IL-8 level in gastric mucosa was improved in both groups but the improvement in the treated group was better. Cenclusien： JQH could effectively treat gastric ulcer and partly reduce its recurrence through improving patients＇ immune function.

Influences of AMY1 gene copy number and protein expression on salivary alpha-amylase activity before and after citric acid stimulation in splenic asthenia children

OBjECTIVE:To compare the correlations between salivary alpha-amylase(sAA) activity and amylase,alpha 1(salivary) gene(AMY1) copy number or its gene expression between splenic asthenia and healthy children,and investigate the reasons of attenuated sAA activity ratio before and after citric acid stimulation in splenic asthenia children.METHODS:Saliva samples from 20 splenic asthenia children and 29 healthy children were collected before and after citric acid stimulation.AMY1 copy number,sAA activity,and total sAA and glycosylated sAA contents were determined,and their correlations were analyzed.RESULTS:Although splenic asthenia and healthy children had no differences in AMY1 copy number,splenic asthenia children had positive correlations between AMY1 copy number and sAA activity before or after citric acid stimulation.Splenic asthenia children had a higher sAA glycosylated proportion ratio and glycosylated sAA content ratio,while their total sAA content ratio and sAA activity ratio were lower compared with healthy children.The glycosylated sAA content ratio was higher than the total sAA content ratio in both groups.Splenic asthenia and healthy children had positive correlations between total sAA or glycosylated sAA content and sAA activity.However,the role played by glycosylated sAA content in sAA activity in healthy children increased after citric acid stimulation,while it decreased in splenic asthenia children.CONCLUSION:Genetic factors like AMY1 copy number variations,and more importantly,sAA glycosylation abnormalities leading to attenuated sAA activity after citric acid stimulation,which were the main reasons of the attenuated sAA activity ratio in splenic asthenia children compared with healthy children.