Objective To investigate the effect of MRE11 on the proliferation and apoptosis of esophageal squamous cancer cells and its molecular mechanism. Methods MRE11 expression was downregulated by MRE11 siRNA transfection in esophageal squamous cancer cells. The AKT agonist SC79 (0, 0.1, 0.5, 1, 1.5, 1.8, 2 μg/ml) were used to treat cells with MRE11 inhibition for 24 h. Overexpression vector pcDNA.3.1-c-myc was constructed and co-transfected cells with MRE11 siRNA. Western blot method was used to detect the protein expressions of MRE11, p-AKT and c-myc in esophageal squamous cancer cells Ec9706 and TE-1. The Annexin-V FITC/PI kit was used to detect the apoptosis of Ec9706 and TE-1 cells; the activity of caspase-3 was detected by the Caspase-3 activity detection kit; the proliferation of Ec9706 and TE-1 cells was tested by the BrdU method. Results The protein expressions of MRE11 in Ec9706 and TE-1 cells were significantly increased, compared with human esophageal epithelial Het-1A cells. After MRE11 siRNA transfection, AKT phosphorylation and the protein expressions of MRE11 and c-myc were significantly decreased in esophageal squamous cancer cells. MRE11 inhibition significantly promoted the apoptosis and caspase-3 activity in Ec9706 and TE-1 cells, while inhibited the proliferation of Ec9706 and TE-1 cells. SC79 (1.5, 1.8 and 2 μg/ml) significantly increased AKT phosphorylation in MRE11-suppressed esophageal squamous cancer cells, and reversed the inhibitory effects of MRE11 inhibition on c-myc protein expression and cell proliferation and the promoting effect on cell apoptosis. Overexpression of c-myc inhibited the inhibitory effect of MRE11 down-regulation on cell proliferation and the promotion on caspase-3 activity. Conclusion MRE11 inhibition could effectively inhibit the proliferation of esophageal squamous cancer cells and promote cell apoptosis by regulating AKT and c-myc.

This study was aimed to investigate the mechanism of all-trans retinoic acid (ATRA) up-regulating apelin expression in vascular smooth muscle cells (VSMCs). The effect of ATRA on apelin expression in the VSMCs was investigated by RT-PCR, real-time PCR and Western blot analysis. To further define whether retinoic acid receptor α (RARα) mediated the induction of apelin by ATRA, endogenous RARα was down regulated by transfection of siRNA against RARα (si-RARα) or RARα was over-expressed by infection of the adenovirus vector pAd-GFP-RARα in the VSMCs. The results showed that ATRA significantly induced apelin expression in a time- and dose-dependent manner in the VSMCs. Although RARα expression was increased in a time-dependent manner, the expressions of RARβ and RARγ were little changed by the ATRA treatment. When VSMCs were treated with a RARα antagonist Ro 41-5253 prior to the addition of ATRA, or si-RARα was used to down regulate endogenous RARα expression, the blockade of RARα signaling partially reduced the response of apelin to ATRA. Moreover, RARα over-expression, induced by infection of pAd-GFP-RARα, further increased the induction of apelin by ATRA. In conclusion, ATRA may up-regulate apelin expression in VSMCs, and the mechanism may be RARα dependent.
Benzoates ; Chromans ; Gene Expression Regulation ; Intercellular Signaling Peptides and Proteins ; metabolism ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, Retinoic Acid ; metabolism ; Retinoic Acid Receptor alpha ; Signal Transduction ; Transfection ; Tretinoin ; metabolism ; Up-Regulation

OBJECTIVE To investigate the effects and mechanism of atractylodin on inflammatory injury of periodontal tissue and alveolar bone loss in periodontitis rats. METHODS A total of 144 SD rats were divided into control group （intragastric and intraperitoneal injection of normal saline）， model group （intragastric and intraperitoneal injection of normal saline）， atractylodin low-dose， medium-dose and high-dose groups （intraperitoneal injection of 6.665， 13.33， and 26.66 mg/kg atractylodin）， metronidazole group （positive control group， intragastric injection of 0.05 g/kg metronidazole， intraperitoneal injection of normal saline）， AMD3100 [stromal cell-derived factor-1 （SDF-1）/CXC chemokine receptor-4 （CXCR4） pathway inhibitor] group （intragastric injection of 1 mg/kg AMD3100， intraperitoneal injection of normal saline）， atractylodin high-dose+AMD 3100 group （intraperitoneal injection of 26.66 mg/kg atractylodin， intragastric injection of 1 mg/kg AMD3100）， with 18 rats in each group. Except for the control group， all other groups of rats were inoculated with Porphyromonas gingivalis to construct a periodontitis model. After successful modeling， they were given relevant medicine or normal saline， once a day， for 4 consecutive weeks. The gingival index of rats was detected； the levels of interleukin-6 （IL-6） and tumor necrosis factor α （TNF-α） in rat serum were also determined； alveolar bone resorption， periodontal histopathologic changes and the number of osteoclasts were detected by methylene blue staining， HE staining and TRAP staining， respectively. The expressions of osteoprotegerin （OPG）， receptor activator of NF-κB ligand （RANKL）， SDF-1 and CXCR4 proteins were determined. RESULTS Compared with the control group， serious pathological injury of periodontal tissue was found in the model group， the gingival index， the levels of IL-6 and TNF- α， alveolar bone absorption value， the number of osteoclasts， and the expression of RANKL protein were all increased significantly （P＜0.05）， while the expressions of OPG， SDF-1 and CXCR4 proteins were decreased significantly （P＜0.05）. Compared with the model group， pathological injury of periodontal tissue in rats was reduced； the gingival index， the levels of IL-6 and TNF-α， alveolar bone resorption value， osteoclast number and RANKL protein expression were decreased significantly， while protein expressions of OPG， SDF-1 and CXCR4 were increased significantly in atractylodin low-dose， medium-dose and high-dose groups and metronidazole group （P＜0.05）. The change trend of corresponding indexes in the AMD3100 group was opposite to the above （P＜0.05）. AMD3100 attenuated the inhibitory effect of high-dose atractylodin on inflammatory response and alveolar bone loss in rats with periodontitis （P＜0.05）. CONCLUSIONS Atractylodin may improve the inflammatory response and alveolar bone loss in periodontitis rats by activating the SDF-1/CXCR4 signaling pathway.

Anti-Inflammatory Agents ; therapeutic use ; Antioxidants ; therapeutic use ; Diabetes Mellitus, Type 2 ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Herbal Medicine ; methods ; Humans ; Hypoglycemic Agents ; therapeutic use

Objective: To evaluate the expression of leucine zipper tumor suppressor 2 (LZTS2) in human breast cancer tissues and cell lines, and to investigate the effects and mechanisms of LZTS2 over-expression on proliferation, invasion and epithelial-mesenchymal transition (EMT) of breast cancer cells. Methods: Fifty pairs of cancerous tissues and para-cancerous tissues resected from breast cancer patients in Department of Breast Surgery of Kaifeng Central Hospital from January, 2016 to December, 2016, as well as breast cancer cell lines (MCF-7, MDA-MB-231, MDA-MB-468) and normal mammary epithelial HBL-100 cells were collected for this study; and Real-time quantitative PCR (qPCR) and Western blotting were used to determine the mRNA and protein expressions of LZTS2 in collected tissues and cell lines. MCF-7 cells were transfected with pcDNA-LZTS2 or pcDNA3.1 (negative control) using lipofectamineTM 2000, and the protein expression of LZTS2 at 49-72 h after transfection was measured by Western blotting; Then, the effects of LZTS2 over-expression on proliferation, migration and invasion of MCF-7 cells were detected by MTT assay and Transwell assay, respectively; Furthermore, Western blotting was performed to detect the expressions of EMT associated proteins (Cyclin D1, Vimentin, Ncadherin, E-cadherin) and PI3K/AKT signaling pathways-related molecules. Results: The mRNA and protein expressions of LZTS2 were down-regulated in breast cancerous tissues and cell lines (MCF-7, MDA-MB-468 and MDA-MB-231) as compared with paired para-cancerous tissues or normal mammary epithelial HBL-100 cells (P<0.05 or P<0.01). Compared with and blank control or pcDNA3.1 group, the protein expression of LZTS2 in MCF-7 cells of pcDNA-LZTS2 group significantly increased (P<0.01), while the proliferation, migration and invasion of MCF-7 cells significantly reduced (P<0.05 or P<0.01). In addition, forced expression of LZTS2 significantly down-regulated the protein expressions of Cyclin D1, Vimentin and N-cadherin (P<0.05 or P<0.01) but up-regulated the expression of E-cadherin in MCF-7 cells (P<0.01), indicating LZTS2 over-expression suppressed PI3K / AKT signaling pathway through inhibiting the expression p-PI3K and p-AKT. Conclusion: The findings collectively demonstrated that the expression of LZTS2 was decreased in breast cancer, and over-expression of LZTS2 efficiently inhibited the proliferation, migration and invasion of breast cancer cells, which might be related with the suppression of PI3K/AKT signaling pathway involved in EMT.

This study was designed to explore the effect of apigenin (Api) on dendritic cell (DCs) maturation and function in murine spleen cells. The single spleen cell was isolated, and then cultured with lipopolysaccharide (LPS) in the present and absence of apigenin. After 24 h, the toxicity of Api and the T cell proliferation were determined by CCK8 kit. In addition, we collected the cell-free supernatants to measure cytokine production using ELISA, collected the cells to determine the DC maturation using flow cytometry. Finally, we purified Api and/or LPS-treated CD11c⁺ DCs which were pulsed with ovalbumin (OVA)_323-339 and then were adoptive transferred into C57BL/6 mice to detect the OVA_323-339-specific T cell proliferation and T helper (Th1) and Th2 cell secreting IFN-γ and IL-4 production, respectively. We found that Api did not affect splenocyte viability, but inhibited the production of pro-inflammatory cytokine IL-1β, IL-6 and TNF-α, not anti-inflammatory cytokine IL-10. In addition, Api inhibited the expression of co-stimulatory CD80, CD86 and MHCII of CD11c⁺ DCs. Finally, compared to LPS+OVA DCs group, DCs from Api and LPS co-treated splenocytes (Api+LPS+DCs) impaired OVA_323-339-specific T cell proliferation and the production of IFN-γ and IL-4 in CD4⁺ T cells, which had the similar responses with OVA+DCs. These data suggest that Api exhibits anti-inflammatory properties via inhibiting DC activation and function, as a new immune-modulator, which may induce immune-tolerance with a benefit to those with chronic inflammation.

Objective:To observe the efficacy of modified Simotang in treatment of adult functional constipation (Qi-stagnancy constipation), and investigate its effects on serum levels of intestinal neurotransmitter nitric oxide synthase (nNOS), nitric oxide (NO), and vasoactive intestinal peptide (VIP), as well as superoxide dismutase (SOD), malonaldehyde (MDA), and glutathione (GSH) levels. Method:One hundred and ten patients with functional constipation were selected and randomly divided into control group (55 cases) and treatment group (55 cases) by referring to random number table. The patients in control group were given with routine therapy, Domperidone tablet (1 tablet/time, tid), and Phenolphthalein tablets (100 mg/time, bid). The patients in treatment group were treated with modified Simotang, 1 dose/day. Both groups were treated for 4 weeks. Then the scores of main clinical symptoms of functional constipation, scores of Qi-stagnancy constipation and clinical efficacy were compared between two groups. Constipation recurrence rate was compared between two groups after stopping medicine. Serum levels of intestinal neurotransmitters nNOS, NO and VIP as well as SOD, MDA, GSH levels were detected in both groups. Result:After treatment, scores for main clinical symptoms (difficult defecation, abdominal distension, defecation time, number of defecation times) and Bristol scores in treatment group were obviously lower than those in control group (P<0.01). Scores for symptoms of Qi-stagnancy constipation (ungratifying defecation, abdominal distension,bowel ringing, frequent flatus, chest and flank tightness) in treatment group were obviously lower than those in control group after treatment (P<0.01). Total clinical efficacy in treatment group 98.04% was superior to that in control group 84.62% ( P<0.05). Constipation recurrence rate was 3.92% and 8.16% in treatment group after 4 and 8 weeks of medication stopping, obviously lower than those in control group 18.18% and 27.78% (P<0.05). After treatment, serum levels of intestinal neurotransmitters nNOS, NO, VIP in treatment group were remarkably lower than those in control group (P<0.01). After treatment, serum levels of SOD and GSH in treatment group were higher while MDA level was lower than those in control group (P<0.01). Conclusion:Based on routine therapy, modified Simotang in treatment of adult functional constipation (Qi-stagnancy constipation) can improve clinical symptoms and syndrome symptoms, increase the clinical efficacy, decrease recurrence rate, and regulate levels of intestinal neurotransmitters nNOS,NO and VIP as well as SOD, MDA, GSH levels.

OBJECTIVETo study the effect of stigma maydis polysaccharide (SMPS) on gastrointestinal movement.

METHODTaking charcoal as the indicator and taking ratio of charcoal movement, beginning time of black excretion and stool amount as the index to observe the effect of SMPS on intestinal movement in mice. Taking emthylorange as the indicator and taking the ratio of residual rate of methylorange as the index to observe the effect of SMPS on gastric emptying in mice. Taking methylene blue as the indicator and taking the time of gastric emptying and movement speed of intestinal content as the index to observe the effect of SMPS on gastrointestinal movement in rats. Observing the changes of cholecystokinin (CCK) level in plasm in rats.

RESULTCompared with control, the ratio of charcoal movement increased in mice (P <0.01). The beginning time of black excretion shortened and the stool amount increased in mice (P <0.01). The ratio of residual rate of methylorange increased in mice (P <0. 01). The time of gastric emptying prolonged in rats (P <0.01). The movement speed of intestinal content in rats accelerated (P <0.01). CCK level in plasm increased in rats (P < 0.05).

CONCLUSIONEffects of stigma maydis polysaccharide on gastrointestinal movement are probably related to the increasing of CCK level in plasm.

Animals ; Cholecystokinin ; blood ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Gastric Emptying ; drug effects ; Gastrointestinal Agents ; isolation & purification ; pharmacology ; Gastrointestinal Motility ; drug effects ; Intestine, Small ; physiology ; Male ; Mice ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar ; Zea mays ; chemistry

Objective To evaluate whether immunization with △A146 Ply could confer protections against pneumococcal infections in murine models and to reveal the possible role of △A146 Ply-specific IgG in the protection elicited.MethodsBALB/c mice were immunized intraperitoneally with △A146 Ply or PBS plus alum.Fourteen days after the third immunization,mice were intranasally challenged with serotype 14 and 19F Streptococcus pneumoniae.Three days after inoculation,lungs were removed from mice and homogenized in PBS,followed by plated on red cell plates.Viable bacteria were counted after overnight incubation.As to the sepsis models,vaccinated mice were challenged intraperitoneally with different dosage bacteria of D39 and serotype 3 strain.The numbers of CFU log10 were compared by Mann Whitney U test and survival rates were analyzed using log-rank test.Passive protection was used to evaluate the role of △A146 Ply-specific IgG in the protection against otherwise lethal infection resulted from D39.Results ELISA analysis demonstrated higher titer specific antibody responses to △A146 Ply was produced after 3 times immunization.Mice boosted twice with △A146 Ply survived significantly longer than that for mice boosted once with △A146 Ply in Alum adjuvant,which were significantly longer than that of control group.Immunization with △A146 Ply was effective in reducing the numbers of pneumococcal strain 31614(serotype 14) and 31693(serotype 19F),which resulted in 50-and 20-fold decreases in bacterial load in the lungs respectively when compared to control protein-immunized mice.60% of vaccinated mice survived the infection with pneumococcal D39 of 1200 CFU.60% protection was achieved when mice intraperitoneally infected with D39 and received △A146 Ply-specific IgG,whereas no mice survived the infection when they were passively administered with △A146 Ply-specific IgG depleted antisera.Conclusion Immunization with △A146 Ply could confer protection against pneumococcal infections,and protection elicited was mediated by △A146 Ply-specific IgG.

Objective To evaluate mechanical radial scanning endoscopic ultrasonography (EUS) for preoperative tumor and lymph node ( TN ) staging of esophageal cancer. Methods From January 2010 to June 2010, a total of 60 patients with esophageal cancer underwent preoperative staging with mechanical radial scanning EUS. The findings of EUS were compared with postoperative pathological outcomes. Results EUS accurately predicted T stage in 80 % of cases and N stage in 71% cases. Sensitivities to T1 , T2 , T3 and T4 were 75% , 100% , 96% and 50% , respectively, and those to N0 and N1 were 55% and 100% , respectively. With exclusion of 11 patients with un-passable lesions, the accurate rate of EUS in T staging of focal and advanced cancers was 90% ( 44/49 ). Conclusion Mechanical radial scanning EUS can accurately predict T and N stages in preoperative patients with esophageal cancer, which also exhibits high differential accuracy in focal and advanced esophageal cancer.