Objective: To screen the flavonoid constituents and targets of Litchi Semen in the intervention of progression and metastasis of colon adenocarcinoma (COAD). Methods: Through DRAR-CPI and SWISS database, potential targets of 19 flavonoids in Litchi Semen were searched. COAD gene expression data and clinical characteristic data from TCGA database were downloaded. Weighted gene co-expression network analysis (WGCNA) was used to establish the gene co-expression network and identify the co-expression module of COAD. The common targets of co-expression module and potential targets were used as the compound to interfere with the drug target of COAD. Protein interaction network analysis, KEGG and GO analysis were performed by String database. The Hub gene was extracted as potential biomarkers of COAD by the cytoHubba, and the interaction network of components, targets and pathways was established by the Cytoscape. The expressions of potential biomarkers were verified by HPA database, and the compounds were docked with the potential biomarkers. Results: A total of 18 co-expression modules were identified with seven of them were correlated with clinical features, such as survival time and tumor stage. Turquoise module was related to the development and transfer of COAD. 19 flavonoids in Litchi Semen acted on 380 potential targets. 34 targets repeated with turquoise module were selected as targets. GO analysis showed that the target points were enriched in 304 GO items, including 229 biological processes, 31 cell composition and 44 molecular functions; KEGG analysis showed that target points were enriched in cancer pathways, cell cycle, and progesterone-mediated 40 pathways including oocyte cancer pathway, cell senescence, and p53 signaling pathway. The genes of CDC25A, CDC25C, CCNB2 and AURKB were screened by cytoHubba as potential biomarkers which related to the progress and transfer of COAD. Compared with para-cancerous tissues, immunohistochemistry results obtained from HPA database showed that the protein expressions of CDC25C, AURKB and CCNB2 in COAD were increased significantly (P < 0.05), which were consistent with gene expression in TCGA data set. Narirutin, procyanidin A2, phloridzin and ent-epicatechin which were well combined to CDC25A, CDC25C and AURKB through hydrogen bond were screened. Conclusion CDC25A, CDC25C, CCNB2 and AURKB were the potential biomarkers closely related to the progression and metastasis of COAD. The mechanism of intervention of flavonoids in Litchi Semen on the progression and metastasis of COAD may be related to the regulation of biological processes, such as cell division, G2/M phase transformation of cell cycle, and the regulation of cancer pathway, p53 signaling pathway and other signaling pathways. Narirutin, procyanidin A2, phloridzin, ent-epicatechin and rutin could be treated as potential inhibitors of CDC25A, CDC25C and AURKB.

Actins ; metabolism ; Adult ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Calcium-Binding Proteins ; metabolism ; Cell Differentiation ; Desmin ; metabolism ; Diagnosis, Differential ; Female ; Fibroadenoma ; metabolism ; pathology ; surgery ; Hamartoma ; pathology ; Humans ; Hyperplasia ; Leiomyoma ; pathology ; Microfilament Proteins ; metabolism ; Muscle, Smooth ; pathology ; Phyllodes Tumor ; pathology

Objective Through observing the clinical effect and the changes of gastrointestinal hormones caused by the treatment of different moxibustion time of the superficial gastritis patients due to the spleen and stomach weakness,to investigate the regulation of the dose-effect relation and the adjustment function on gastrointestinal hormone of serum of patients caused by the warming and nourish effect with moxibustion.Methods Eighty-four superficial gastritis Patients due to the spleen and stomach weakness were divided into group 1(treated by warming moxibustion for twenty minutes)(n=28),group 2(treated by warming moxibustion for forty minutes)(n=28),and drug(n=28)groups.The changes of the content of the Prostaglandin E2(PGE2)、Somatostatin(SS)and Epidermal growth factor(EGF)were observed before and after treatment between 3 groups and the clinical effect in different time.Results ①All 3 groups were compared after treatment,their clinical effect had no significant difference(P＞0.05):②Compared with pre-treatment,the superficial gastritis symptom score had a very significant difference after the treatment(3.07±1.54)、(3.11±1.40)、(3.79±2.25)and during the 1 month follow-up(2.25±1.32)、(2.57±1.10)、(4.11±2.48),(P＜0.01);③After treatment,the content of the PGE2 of the serum was increased obviously,compared with pre-treatment.There was a significant difference in each group(33.751±1.267)pg/ml、(33.774±8.583)pg/ml、(32.583±8.259)pg/ml,(P＜0.05);After warming moxibustion for forty minutes,the content of the EGF of the serum was increased obviously,compared with pre-treatment,showing a significant difference(1.331±0.823)pg/ml,(P＜0.05).Conclusion ①All of the three treatment methods had significant curative effect,and the curative effect had no significant difference among these 3 groups.But the moxibustion groups were markedly higher than the drug group in long-term result;②The moxibustion had established adjustment function on gastrointestinal hormone of the superficial gastritis patients due to the spleen and stomach weakness,which suggested that the gastrointestinal hormone may participate in its onset and the process of pathology and physiology.

Objective: To observe the effects of different doses of ginger-partitioned moxibustion on serum trefoil factor 1 (TFF1) and mucin 5AC (MUC5AC) levels, as well as the expression of epidermal growth factor receptor (EGFR) in gastric mucosa of rats with spleen deficiency syndrome, therefore, to explore the possible mechanism and the dose-effect characteristics of ginger-partitioned moxibustion in spleen deficiency syndrome. Methods: Seventy-five SPF grade Sprague-Dawley (SD) rats were randomly divided into a blank control group (group A), a model group (group B), a 3 moxa-cone ginger-partitioned moxibustion group (group C1), a 6 moxa-cone ginger-partitioned moxibustion group (group C2) and a 9 moxa-cone ginger-partitioned moxibustion group (group C3) using random number table method, 15 rats in each group. Except group A, rats in the other groups received intragastric administration of 4 ℃200% concentrated Da Huang (Radix et Rhizoma Rhei) to prepare spleen deficiency syndrome model. After successful modeling, rats in group B received no treatment; rats in group C1, C2 and C3 were treated with 3, 6 and 9 moxa-cone ginger-partitioned moxibustion at Zusanli (ST 36) and Zhongwan (CV 12) respectively for 8 continuous days. The general symptom score of rats was observed. The serum levels of TFF1 and MUC5AC were detected by enzyme-linked immunosorbent assay (ELISA). The expression of EGFR protein in gastric mucosa was detected by immunohistochemistry. Results: After the treatment, compared with group A, the spleen deficiency symptom score was increased in group B, the levels of serum TFF1 and MUC5AC, the EGFR protein expression in gastric tissues of group C1, C2 and C3 were significantly increased (all P<0.01); compared with group B, the spleen deficiency scores were decreased in group C1, C2 and C3, and the serum levels of TFF1 and MUC5AC, as well as EGFR protein expression in gastric tissues were increased (all P<0.01). Compared with group C1, the spleen deficiency scores were decreased in group C2 and C3, the serum levels of TFF1 and MUC5AC, and the expression of EGFR protein in gastric tissues were increased (all P<0.01), however, there was no significant difference between group C2 and C3 (all P>0.05). The mechanism may be related to the increase of serum TFF1 and MUC5AC levels and activation of EGFR protein. Conclusion: Ginger-partitioned moxibustion can improve the symptoms, as well as promote the proliferation and repair of gastric mucosa in rats with spleen deficiency. The therapeutic efficacy of 6 or 9 moxa-cone ginger-partitioned moxibustion is better than that of 3 moxa-cone ginger-partitioned moxibustion, while the efficacies are equivalent between 6 and 9 moxa-cone ginger-partitioned moxibustion groups.

Objective To observe effects of different dosages of moxibustion with ginger-separated moxibustion on expressions of mitogen extracellular kinase (MEK) 1/2 and extracellular regulated protein kinase (ERK) 1/2 of gastric tissue in rats with spleen deficiency; To explore the possible mechanism and the dose-effect relationship. Methods Seventy-five SD rats were randomly divided into blank control group, model group, ginger-separated moxibustion for three zhuang group, six zhuang group and nine zhuang group according to random digits table method, with fifteen rats in each group. The rat model of spleen deficiency was established by intragastric administration with 200% Rhei Radix et Rhizoma infusion at 4 ℃. Ginger-separated moxibustion groups were treated with different dosage of moxibustion at "Zusanli", "Zhongwan" for eight days after the modeling. Pathological changes of gastric tissue by HE staining were observed under light microscope, and immunohistochemistry was used to detect the expressions of MEK1/2 and ERK1/2 protein in gastric tissue of rats. Results Compared with the blank control group rats, the gastric mucosa injury in the model group was obvious, which showed that the damage and abscission was more serious; compared with the model group, the gastric mucosa of rats was partly exfoliated and the damage was improved in three zhuang group, and the surface of gastric mucosa of rats was more complete and damage was improved obviously in six zhuang group and nine zhuang group; compared with the blank control group, the expressions of MEK1/2 and ERK1/2 protein in gastric tissue increased obviously in other groups (P<0.01);compared with three zhuang group, the expressions of MEK1/2 and ERK1/2 protein in gastric tissue increased in six zhuang group and nine zhuang group (P<0.01), but the effects of the two group were similar, without statistical significance (P>0.05). Conclusion Ginger-separated moxibustion can repair gastric mucosa in rats with spleen deficiency, which may be closely associated with its effect in increasing the expressions of MEK1/2 and ERK1/2 protein in gastric tissue and activating the MEK/ERK signal transduction pathway.

Adenocarcinoma ; metabolism ; pathology ; surgery ; Cardia ; Esophageal Neoplasms ; metabolism ; pathology ; surgery ; Esophagogastric Junction ; metabolism ; pathology ; surgery ; Humans ; Neoplasm Staging ; Receptor, ErbB-2 ; metabolism ; Sirtuin 1 ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; surgery ; Survival Rate

To investigate the molecular mechanism of hydroxycamptothecin (HCPT)-mediated anti-hepatic fibrosis by evaluting its effects on expression of tumor growth factor-beta 1 (TGFb1), alpha-smooth muscle actin (a-SMA) and collagen I (Col I) in hepatic satellite cells (HSCs). Cultured HSCs were treated with different concentrations of HCPT: low-dose group, 0.25 mg/L; middle-dose group, 0.5 mg/L; high-dose group, 0.75 mg/L; and control group, 0 mg/L. Cell proliferation was assessed by the MTT assay. The mRNA expressions of TGFb1, a-SMA and Col I were determined by reverse transcription-polymerase chain reaction. The protein expressions of TGFb1 and a-SMA were detected by Western blot. The content of Col I in the cultured HSCs' supernatant was measured by enzyme-linked immunosorbent assay. The MTT absorbance values of the low-dose group (0.631+/-0.074), middle-dose group (0.469+/- 0.012) and high-dose group (0.204+/- 0.001) were significantly lower than that of the control group (0.793+/-0.098; F = 82.86, P less than 0.01). Compared with the control group, the HCPT-treated groups showed significantly down-regulated gene expressions of TGFb1 (control: 0.716+/-0.064 vs. low: 0.611+/-0.040, middle: 0.510+/-0.014, high: 0.403+/-0.026), a-SMA (control: 0.696+/-0.075 vs. low: 0.579+/-0.037, middle: 0.470+/-0.024, high: 0.299+/-0.017), and Col I (control: 1.019+/-0.056 vs. low: 0.835+/-0.022, middle: 0.696+/-0.055, high: 0.322+/-0.104) (all, P less than 0.01). Meanwhile, HCPT-treated HSCs showed significantly reduced protein expressions of TGFb1 (control: 0.872+/-0.053 vs. low: 0.654+/-0.047, middle: 0.545+/-0.042, high: 0.436+/-0.039) and a-SMA (control: 0.858+/-0.050 vs. low: 0.620+/-0.045, middle: 0.525+/-0.042, high: 0.434+/-0.052) (all, P less than 0.01). The Col I levels secreted by HSCs were significantly lower in the HCPT-treated groups (low: 168.367+/-16.453 ng/ml; middle: 141.284+/-11.731 ng/ml; high: 132.910+/-10.048 ng/ml) than in the control group (188.733 +/-18.299 ng/ml) (all, P less than 0.01). The mechanism of HCPT-mediated anti-hepatic fibrosis may involve down-regulation of TGFb1 expression to inhibit HSC proliferation and activation, as well as reduction of Col I synthesis and secretion.
Actins ; metabolism ; Animals ; Camptothecin ; analogs & derivatives ; pharmacology ; Cell Proliferation ; Cells, Cultured ; Collagen Type I ; metabolism ; Hepatic Stellate Cells ; cytology ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism

BACKGROUNDThe vascular endothelial growth factor (VEGF) is involved in the initiation of retinal vascular leakage and nonperfusion in diabetes. The intracellular adhesion molecule-1 (ICAM-1) is the key mediator of the effect of VEGFs on retinal leukostasis. Although the VEGF is expressed in an early-stage diabetic retina, whether it directly up-regulates ICAM-1 in retinal endothelial cells (ECs) is unknown. In this study, we provided a new mechanism to explain that VEGF does up-regulate the expression of ICAM-1 in retinal ECs.

METHODSBovine retinal ECs (BRECs) were isolated and cultured. Immunohistochemical staining was performed to identify BRECs. The cultured cells were divided into corresponding groups. Then, VEGF (100 ng/ml) and other inhibitors were used to treat the cells. Cell lysate and the cultured supernatant were collected, and then, the protein level of ICAM-1 and phosphorylation of the endothelial nitric oxide synthase (eNOS) were detected using Western blotting. Griess reaction was used to detect nitric oxide (NO).

RESULTSWestern blotting showed that the VEGF up-regulated the expression of ICAM-1 protein and increased phosphorylation of the eNOS in retinal ECs. Neither the block of NO nor protein kinase C (PKC) altered the expression of ICAM-1 or the phosphorylation of eNOS. The result of the Western blotting also showed that inhibition of phosphatidylinositol 3-kinase (PI3K) or reactive oxygen species (ROS) significantly reduced the expression of ICAM-1. Inhibition of PI3K also reduced phosphorylation of eNOS. Griess reaction showed that VEGF significantly increased during NO production. When eNOS was blocked by L-NAME or PI3K was blocked by LY294002, the basal level of NO production and the increment of NO caused by VEGF could be significantly decreased.

CONCLUSIONROS-NO coupling in the retinal endothelium may be a new mechanism that could help to explain why VEGF induces ICAM-1 expression and the resulting leukostasis in diabetic retinopathy.

Animals ; Blotting, Western ; Cattle ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Gene Expression ; drug effects ; Intercellular Adhesion Molecule-1 ; metabolism ; Nitric Oxide ; metabolism ; Reactive Oxygen Species ; metabolism ; Retina ; cytology ; Vascular Endothelial Growth Factor A ; pharmacology

Objective To observe the cost-effectiveness of using continuous subcutaneous insulin infusion (CS Ⅱ) and multi-point daily insulin injections (MDI) in controlling blood sugar in the newly hospitalized type 2 diabetes patients. Methods Retrospective analysis on 86 cases taking CS Ⅱ and 103 cases using MDI on a 'blood sugar control program' among the newly hospitalized patients with type 2 diabetes. The period for observation was 2 weeks, using cost-effectiveness analysis methods to evaluate the two treatment programs. Results After two weeks of treatment, the effectiveness in the control of blood sugar in CS Ⅱ group was similar to the MDI group, with no significant difference(P<0.05) and the adverse reactions were similar. Costs in the CS Ⅱ program (Yuan/person) was less than in the MDI program (1478.34 vs. 1620.46), with significant differences (P< 0.05). The cost-effectiveness ratios (C/E) were 15.07 in the CS Ⅱ group, and 16.34 in the MDI group, with no significant difference (P>0.05). In order to further reduce the cost of CS Ⅱ group as a reference, the incremental cost-effectiveness ratio (△C/ △E)ofthe MDI group was 129.20. Conclusion Costs-effective of the CS Ⅱ program was better than the MDI one in treating the newly hospitalized patients with type 2 diabetes, suggesting that CS Ⅱ program might be a better choice for hospitals to carry on an intensive insulin therapy program.

·AIM: To investigate the correlation of matrix metalloproteinase -9 (MMP-9), glycated albumin (GA), glycosylated hemoglobin ( HbA1c ) and adipokines ( including visfatin, resistin and leptin ) with diabetic retinopathy ( DR) .·METHODS:From March 2015 to March 2017, 74 patients with DR were treated in our hospital, including 40 patients ( 80 eyes ) with non proliferative diabetic retinopathy ( NPDR ) and 34 patients ( 68 eyes ) with proliferative diabetic retinopathy ( PDR ) , and diabetes mellitus 40 patients ( 80 eyes ) with non DR ( NDR ) and 40 healthy volunteers (80 eyes) were selected as controls, the levels of MMP-9, GA, HbA1c, visfatin, resistin and leptin in each group were detected.·RESULTS: PDR group visfatin was 4. 41 ± 0. 82ng/mL, was significantly lower than the NPDR group, NDR group and control group ( P<0. 05 ) , while, resistin, leptin and MMP- 9 were 9. 01 ± 1. 04ng/mL, 17. 96 ± 2. 03μg/L and 740. 06 ± 84. 43μg/L, GA and HbA1c were 26. 14%± 4. 57%and 17. 60% ± 1. 91%, significantly higher than those of NPDR group, NDR group and control group ( P<0. 05 ) . NPDR group visfatin was 6. 44 ± 0. 79ng/mL, was significantly lower than that of NDR group and control group (P<0. 05), while, resistin, leptin and MMP-9 were 7. 80±0. 87ng/ml, 15. 68±1. 98μg/L and 634. 12±80. 22μg/L,GA and HbA1c were 22. 06%± 4. 38% and 12. 46%± 1. 69%, significantly higher than those of NDR group and control group (P<0. 05). MMP-9, GA, HbA1c were positively with DR levels ( rs = 0. 523, 0. 461 and 0. 414, P<0. 05 );visfatin was negatively correlated with DR levels ( rs = -0. 433, P < 0. 05 ), resistin and leptin were positively correlated with DR levels (rs=0. 401 and 0. 460, P<0. 05).·CONCLUSION: MMP-9, GA, HbA1c, and adipokines may play a role in the development and progression of DR, in which MMP-9 is associated with adipokines, both are not significantly related to the levels of GA and HbA1c.