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Objective To study the effect of three kinds of commonly used liquid culture media for in vitro cell experiments on elastic modulus of breast cancer cells, so as to provide references for developing novel diagnosis and treatment approach of tumour based on mechanics principles. Methods The elastic modulus and adhesion force of breast cancer cells MCF7 to atomic force microscopy (AFM) probes in phosphate buffered solution(PBS), Dulbecco’s modified eagle media (DMEM) and DMEM+10% fetal bovine serum (FBS) were measured using AFM technology. Results The elastic moduli of breast cancer cells in PBS, DMEM and DMEM+10% FBS were 2.59, 2.11 and 1.59 kPa, respectively. The cell adhesion forces in the above three kinds of liquid medium environment were 63.81, 66.09 and 121.97 pN, respectively. Cell adhesion force in DMEM+10%FBS was significantly different from that of the other two kinds of liquid media. Conclusions There are significant differences in elastic modulus of breast cancer cells in three kinds of liquid media. The difference between DMEM and DMEM+10%FBS might be caused by the different adhesion force caused by serum proteins in the media, and the difference between DMEM and PBS might be attributed to the difference in pH of the media.
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Background:Previous study has found that harmine inhibited the proliferation of gastric cancer cells by down-regulating cyclooxygenase-2(COX-2)expression. However,its molecular mechanism is not fully clear. Aims:To investigate the effect of harmine on proliferation and apoptosis of gastric cancer cells,and explore the role of PTEN/Akt/MDM2 signaling pathway in this process. Methods:Human gastric adenocarcinoma cell line SGC-7901 and MKN-45 were treated with harmine at different concentrations(2,4,8,16,32 μg/mL)for 24,48,and 72 hours. The cell proliferation and apoptosis were detected by MTT assay and Hoechst staining,respectively. The expressions of PTEN,COX-2, phosphorylated Akt(p-Akt)and p-MDM2 were measured by Western blotting. Results:Harmine dose- and time-dependently inhibited proliferation and induced apoptosis of SGC-7901 and MKN-45 cells. Also,harmine dose-dependently increased PTEN expression,and inhibited p-Akt,p-MDM2 and COX-2 expressions in SGC-7901 and MKN-45 cells. Conclusions:Harmine may inhibit proliferation and induce apoptosis of gastric cancer cells via down-regulating COX-2 expression through PTEN/Akt/MDM2 signaling pathway.
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Background: Previous study has found that ursolic acid (UA) inhibited the proliferation of gastric cancer cells by the down-regulation of cyclooxygenase-2 (COX-2) expression.However,its molecular mechanism is not fully clear.Aims: To investigate the role of adenosine monophosphate-activated protein kinase (AMPK)/signal transducer and activator of transcription 3 (STAT3)/COX-2 signaling pathway in UA-mediated inhibition of gastric cancer cells proliferation.Methods: AMPK-pLVX,AMPK-shRNA,STAT3-pLVX,STAT3-shRNA plasmids were constructed,and then were transfected into human gastric cancer cell lines SGC-7901 and MKN-45,respectively.Gastric cancer cells were cultured with different concentrations of UA for different times.The expressions of phosphorylated AMPK (p-AMPK),phosphorylated STAT3 (p-STAT3) and COX-2 were measured by Western blotting,and cell proliferation was detected by CCK-8 assay.Results: UA dose-and time-dependently increased p-AMPK expression,inhibited p-STAT3 and COX-2 expressions in SGC-7901 and MKN-45 cells.Knockdown of AMPK blocked UA-induced inhibition of STAT3 phosphorylation and COX-2 expression.Overexpression of STAT3 blocked UA-induced down-regulation of COX-2 expression.Knockdown of AMPK and overexpression of STAT3 blocked UA-induced inhibition of proliferation of gastric cancer cells.Conclusions: UA may inhibit the proliferation of gastric cancer cells via down-regulation of COX-2 expression through AMPK/STAT3 pathway.
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Objective To evaluate the relationship of preoperative neutrophil-lymphocyte ratio (NLR) with clinicopathological features and prognosis of colorectal cancer in middle-aged and elderly patients.Methods A retrospective analysis was performed in 212 patients with colorectal cancer in the First Affiliated Hospital of Nanjing Medical University from January 2011 to June 2013.All patients were divided into middle-aged group (46-65 year old,n=130) and old-aged group (66-89 year old,n=82),The optimal cut-off point of NLR was identified by the area under receiver operating characteristic curve,while NLR ＞ 3.13 and NLR≤3.13 were classified as high and low NLR group.The clinicopathological features and prognosis between the two groups were compared.Results There was no difference in gender,tumor growth site,depth of invasion,tumor embolus,lymphatic metastasis,distant metastasis,TNM stage between low and high NLR group (allP＞ 0.05).However,the difference between high NLR group and low NLR group in old-aged group with diabetes mellitus was statistically significant (P＜0.05).The 1-,2-,and 3-year survival rate of the overall 212 patients were 96.2％ (204/212),87.7％ (186/212) and 74.5％ (158/212) In middle-aged group,the 1-,2-,and 3-year survival rates were 98.8％ (85/86),90.7％ (78/86) and 84.9％ (73/86) respectively in low NLR group,but 95.5％ (42/44),84.1％ (37/44) and 72.7％ (32/44) respectively in high NLR group,(allP＜0.05).In old-aged group,the 1-,2-,and 3-year survival rates were 95.7％ (44/46),89.1％ (41/46) and 73.9％ (34/46) respectively in low NLR group,but 91.7％ (33/ 36),83.3％ (30/36) and 52.8％ (19/36) respectively in high NLR group (all P＜0.05).Cox regression showed that TNM stage and NLR were independent risk factors for the prognosis of the middle-aged and elderly patients with colorectal cancer (P＜0.05 or P＜0.01).Conclusions Preoperative NLR ＞ 3.13 suggest that the prognosis is poor in middle-aged and elderly patients with colorectal cancer.
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Aim Our previous study has found that ur-solic acid( UA) increased intracellular reactive oxygen species ( ROS ) production and adenosine monophos-phate-activated protein kinase ( AMPK ) phosphoryla-tion, inhibited signal transducer and activator of tran-scription 3 ( STAT3 ) phosphorylation and cyclooxygen-ase-2 ( COX-2 ) expression in gastric cancer cells . However , the molecular mechanism by which UA in-hibits COX-2 expression in gastric cancer cells has not been fully clarified .In this study we aimed to further clarify the signal transduction pathways involved in the UA-mediated inhibition of COX-2 expression in gastric cancer cells .Methods Human gastric cancer cell lines SGC-7901 and MKN-45 were routinely cultured in RPMI-1640 medium supplemented with 10% heat-in-activated fetal calf serum .Sub-confluent cell cultures were pre-treated with antioxidant N-acetylcysteine ( NAC) , AMPK activator 5-amino-4-imida-zolecarbox-amide-riboside ( AICAR ) , AMPK inhibitor compound C, or STAT3 inhibitor WP1066 and then treated with or without UA for 24 h.The expression of AMPK and phosphorylated AMPK ( p-AMPK ) , STAT3 and phos-phorylated STAT3 ( p-STAT3 ) , as well as COX-2 was detected by Western blot analysis .Results Antioxi-dant NAC and AMPK inhibitor compound C blocked UA-induced inhibition of STAT 3 phosphorylation and down-regulation of COX-2 expression in gastric cancer cells.Both AMPK activator AICAR and UA inhibited STAT3 phosphorylation and COX-2 expression; the combination of two drugs resulted in further reduction . STAT3 inhibitor WP1066 did not affect UA-induced AMPK phosphorylation , whereas it inhibited STAT3 phosphorylation and COX-2 expression .The inhibitory effects on the STAT3 phosphorylation and COX-2 ex-pression were significantly enhanced when SGC-7901 and MKN-45 cells were treated simultaneously with WP1066 plus UA.Conclusion UA inhibits COX-2 expression in gastric cancer cells , which may be medi-ated through ROS/AMPK/STAT3 signal transduction pathway .
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Background:Recent studies have showed that high homocysteine(Hcy)level can increase the risk of gastric cancer, but no related studies have been reported on role of Hcy in gastric precancerous diseases. Aims:To investigate the role of serum Hcy,folic acid and vitamin B12 in patients with gastric cancer and precancerous diseases. Methods:Eighty-six normal controls,46 atrophic gastritis,46 gastric ulcer,31 gastric polyp,52 gastric cancer patients diagnosed by gastroscopy and pathology were enrolled. Serum levels of Hcy,folic acid and vitamin B12 were determined,and their correlations with clinicopathological features in gastric cancer were analyzed. Results:Compared with normal controls, serum Hcy level in patients with atrophic gastritis and gastric cancer was significantly increased(P < 0. 05);serum folic acid and vitamin B12 levels in patients with gastric ulcer,gastric polyp and gastric cancer were significantly decreased(P <0. 05). Serum Hcy level in patients with gastric cancer was positively correlated with depth of tumor infiltration,TNM staging and lymph node metastasis(P < 0. 05),however,serum folic acid and vitamin B12 levels had no correlation with clinicopathological features. Conclusions:Hcy level is increased in chronic atrophic gastritis,gastric cancer;levels of folic acid and vitamin B12 are decreased in gastric ulcer,gastric polyp and gastric cancer. High level of Hcy is involved in infiltration and metastasis of gastric cancer. Intervention in patients with high level of Hcy,low levels of folic acid and vitamin B12 might be an effective strategy for the prevention and treatment of gastric cancer and precancerous diseases.
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Objective To evaluate the clinical application of enteral nutrition by nasojejunal tube insertion and by percutaneous endoscopic gastrostomy (PEG) in elderly patients.Methods A total of 65 elderly patients with dysphagia recruited at our department from January 2010 to November 2014 were divided into the nasojejunal tube feeding group (35 cases) and the PEG feeding group (30 cases).Differences between these two groups in nutritional indexes,immunological indexes,complications and mortality were analyzed retrospectively.Results Serum total protein,albumin and prealbumin and upper arm circumferences all increased after treatment with nasojejunal tube feeding or percutaneous endoscopic gastrostomy (P＞0.05).There was overall improvement in nutritional status,as assessed by Nutritional Risk Screening 2002 (NRS2002).Specifically,the before/one month-after-treatment ratio of scores was 3.72±0.91/1.90±0.61 (t=7.24,P＜0.01) for the nasojejunal tube feeding group and 3.52±1.23/2.02±0.53 (t=4.17,P＜0.01) for the PEG feeding group.Compared with NRS2002 scores at one month post-operation,further improvement was achieved at 3 months postoperation both for the nasojejunal tube feeding group (1.89±0.65,t=5.21,P＜0.01) and for the PEG feeding group (1.91±0.62,t=4.40,P＜0.01).There was no difference in the indexes of nutrition,immune status or mortality between the two groups (P＞0.05).Although improvement in CD3+,CD4+,CD8+,CD4+/CD8+,IgA,IgG,and IgM was seen in both groups after operation,the differences did not reach statistical significance (P＞0.05).The incidence of aspiration pneumonia was notably lower (P＜0.05) while the incidence of diarrhea was much higher (P＜0.05) in the nasojejunal tube feeding group than in the PEG feeding group at one month and three months.The two groups had similar causes of death and mortality rates.Conclusion Both nasojejunal tube and PEG feeding can improve the nutritional status of elderly patients with dysphagia.However,the choice for the route of nutrition should be individualized.
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Objective To investigate the value of detecting methylated SEPT9 gene (mSEPT9) in serum and stool in screening colorectal cancer in elderly people.Methods Subjects were divided into three groups, the colorectal cancer group (n=82), the colorectal polyps group (n=80) and the healthy control group (n=100).Real-time quantitative polymerase chain reaction (PCR) was used to detect mSEPT9 in peripheral blood and feces.The relationship between mSEPT9 and clinical and pathological characteristics of colorectal cancer was analyzed.The agreement between serum and stool results was analyzed.Results The positive rate of serum mSEPT9 was 73.2％ in the colorectal cancer group, 6.3％ in the colorectal polyps group and 4.0％ in the healthy control group, with statistical significance between the colorectal cancer group and the other two groups (P＜0.01).The sensitivity and specificity for plasma mSEPT9 detection in screening colorectal cancer were 73.2 ％ and 95.6％, respectively.The positive rate was higher in patients with tumor maximum diameter over 3.0 cm, invasion of the entire serosa layer, lower differentiation or advanced clinical staging (P＜0.05).The detection results of plasma mSEPT9 were in complete agreement with those of stool mSEPT9 from the same patients.Conclusions The expression of mSEPT9 is high in elderly patients with colorectal cancer and has a high sensitivity and specificity for PCR detection.Complete agreement can be seen in results from plasma and feces.Detecting mSEPT9 can be a valuable approach to screening colorectal cancer in the elderly.
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BacKground:The deveIopment of acute pancreatitis( AP ) is often accompanied by pancreatic microcircuIation disturbance and aIterations in bIood coaguIation and fibrinoIytic systems, which resuIts in abnormaIities of pIateIet parameters. Aims:To investigate the changes of pIateIet parameters in eIderIy AP patients and the effect of uIinastatin ( UTI)on pIateIet parameters and disease status. Methods:Two hundred and seventeen eIderIy AP patients(≥60 years) were enroIIed retrospectiveIy from Aug. 2009 to Dec. 2013 at the First AffiIiated HospitaI of Nanjing MedicaI University. According to the severity of the disease,patients were divided into three groups:miId AP( MAP),moderateIy severe AP ( MSAP),and severe AP( SAP). The pIateIet parameters,incIuding PLT,MPV and PDW before and after treatment,as weII as the differences in pIateIet parameters and cIinicaI efficacy between conventionaI therapy and UTI therapy were compared and anaIyzed. Results:In MSAP and SAP groups,PLT at admission was significantIy Iower(P<0. 01),whiIe MPV and PDW were significantIy higher(P<0. 05)than those in MAP group. After 1-week treatment,PLT increased significantIy(P<0. 01)and MPV and PDW decreased significantIy(P<0. 05)in MSAP and SAP groups. UTI therapy was superior to conventionaI therapy in increasing PLT( P <0. 01 ),decreasing MPV and PDW( P <0. 01 ),and improving the overaII efficacy in MSAP and SAP groups( MSAP:92. 3% vs. 81. 2%,P <0. 01;SAP:90. 0% vs. 80. 8%,P<0. 01). Conclusions:Changes of pIateIet parameters in eIderIy MSAP and SAP patients are different from those in MAP patients,which might refIect the disease severity. UTI is effective in reducing the activity of pIateIet and can be used in the treatment and prevention of disease progression in eIderIy AP patients.
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Objective: To establish the method of percutaneous endoscopic gastrostomy(PEC) and percutaneous endoscopic jejunostomy (PEJ) for enteral nutrition. Methodes: PEG tubes were placed in 114 patients with Pull method. On the foundation of PEG, PEJ tubes were placed in 26 patients by pushing endoscopy to send tubes through Treitz ligment with usingthe the clip. Results: All PEG insertion was performed successfully. PEJ tubes were placed successfully with a new method in 26 patients. 15 patients had a little blooding and 8 patients had slight infection. 21 patients had respiratory tract infection and had been cured by using antibiotic. There was no severe complication. Conclusion: PEG is simple、safe、efficient. The new method of PEJ is feasible.
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Objective Cyclooxygenase (COX), the key enzyme for synthesis of prostaglandins (PGs), exists in two isoforms (COX 1 and COX 2). Conventional non steroidal anti inflammatory drugs (NSAIDs) inhibit both COX 1 and COX 2 activities and induce serious gastrointestinal side effects. Specific COX 2 inhibitors are expected to cause fewer gastric side effects. The aim of this study was to investigate the effects of specific and non specific COX 2 inhibitors on gastric wound healing following acid induced injury. Methods Male Sprague Dawley rats were given 1 ml of 0.6 mol/L hydrochloric acid (HCl) into the stomach. Levels of COX 1 and COX 2 in gastric mucosa were analyzed using western blotting and immunohistochemical staining. At 10 minutes after the administration of the acid, the animals were given 0.4, 4 and 40 mg/kg of NS 398 (NS) or 40 mg/kg of indomethacin (IM). Control group was given 1% arabic gum (AG) in a volume of 5 ml/kg. The rats were sacrificed and laparotomized before and at 1, 3, 6, 12, 24, 48 h after acid administration. Lesion index (LI) was measured and morphological changes of gastric mucosa were assessed under light microscopy. Results Expression of COX 2 was enhanced mainly in surface epithelial cells and neck cells after HCl administration. NS and IM delayed the healing of gastric injury. At 12 h after acid administration, LI was (1.42 ? 0.23)% and (1.42 ? 0.29)% in the groups treated with 4 and 40 mg/kg of NS respectively, and (1.62 ? 0.44)% in the group treated with 40 mg/kg of IM, which was significantly higher than that in control group [(0.58?0.24)%, P
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Objective To investigate the clinical and histopathologic features of patients with primary gastrointestinal malignant lymphoma ( PGIML). Methods The clinical and histopathologic data of 22 patients with PGIML were reviewed and analyzed retrospectively. All cases were confirmed with histological specimen obtained from endoscopic biopsies or surgery. Results Abdominal pain was the most common presenting symptom, seen in 15 of 22 patients (68.2% ). The incidence of PGIML was highest in stomach, seen in 12 of 22 patients (54. 5% ). Modularity of the mucosal surface was the most common endoscopic finding, seen in 15 of 21 patients (71. 4% ). The positive rate of endoscopic biopsy for the diagnosis of PGIML was 52. 6% (10/19 biopsy cases). All cases were non-Hodgkins lymphomas ( NHL). Twenty cases were muco-sa associated lymphoid tissue (MALT) lymphoma, and 13 of 20 cases were extra-nodal marginal zone B-cell lymphoma of MALT. Conclusions Abdominal pain is the most common symptom and the stomach was the most common location in PGIML. Extra nodal marginal zone B-cell lymphoma of MALT is the main histopathologic feature. The prognosis of PGIML is related to the surgical procedure and the post operative chemotherapy.
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Objective To evaluate the accuracy of RUT for the diagnosis of H. pylori infection before and after eradication therapy using bacterial culture and histological examination as gold standard. Methods A total of 250 patients undergoing gastroendoscopy, including 127 and 123 patients with or without previous H. pylori eradication therapy respectively were randomly selected. Each of the biopsy specimens was taken from the antrum and body of stomach for RUT, culture, and histological examination. The data of positive culture and/or positive histology were defined as H. pylori infection. It was judged H. pylori negative or eradication of the bacteria only when the results of culture and histological examination were both negative. Results In cases before eradication therapy, RUT correctly diagnosed in 84 of 86 H. Pylori-positive cases, and 34 of 37 H. Pylori-negative cases. The overall accuracy was 95.9%. The sensitivity and specificity of RUT before eradication were 97. 7% and 91. 9% respectively. In cases after eradication therapy, RUT gave correct diagnoses in 18 of 28 H. pylori - positive cases and in 98 of 99 H. pylori-negative cases with sensitivity 64. 3% and specificity 99. 0% . However, when RUT was used more than 6 months after the end of eradication therapy, both the sensitivity and specificity increased to 100%. Conclusions RUT is highly accurate for the diagnosis of H. pylori infection in untreated and treated patients who were examined more than 6 months after the completion of therapy.
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Objective Adenocarcinoma of the stomach has been established as a consequence of gastric infection with Helicobacter pylori(H.pylori).The specific role of H.pylori in the pathogenesis is unknown yet.Recent studies indicate that expression of mitogen inducible cyclooxygenase-2(COX-2) occurs in gastrointestinal tumors.The purpose of the present study was to investigate expression of COX-2 protein in the human stomach with or without H.pylori infection.Methods Twenty-seven subjects who were asymptomatic referred to the hospital for healthy examination including endoscopic screening.Biopsy specimens were obtained from the subjects without any macroscopic lesions,such as peptic ulcer or gastric malignancies.H.pylori infection was determined by rapid urease test(CLO test),bacterial culture and histology(Giemsa staining).Expression of COX-2 was evaluated by immunohistochemistry using the avidin-biotin-peroxide complex(ABC) method The association between COX-2 expression and H.Pylori infection was assessed by Fisher's exact test.A P value less than 0.05 was considered to be statistically significant.Results specific immunostaining for COX-2 was observed in antral mucosa of 18 subjects infected with H.pylori COX-2 was expressed in gastric mucosal epithelia,mainly in the foveolar epithelial cells.Furthermore,COX-2 was also observed in the neck cells of the gastric glands and inflammatory mononuclear cells beneath the mucosal epithelia.Expression of COX-2 was never found in the gastric mucosa of H.pylori-negative subjects.A positive association of H.pylori infection with COX-2 expression was statistically significant(P
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AIM: To clarify the effects of specific and non-specific cyclooxygenase-2 (COX-2) inhibitors on gastric epithelial cell proliferating and gastric healing following acid-induced damage. METHODS: Male Sprague-Dawley rats were given 1 mL of 0.6 mol/L hydrochloric acid (HCl) into the stomach. Ten minutes after the administration of the acid,the animals were given NS-398 (COX-2 inhibitor) or indomethacin. Levels of COX-1 and COX-2 in the gastric mucosa before and after HCl-administration were analyzed using western blotting and immunohistochemical staining. Proliferating cell nuclear antigen (PCNA) was detected using immunohistochemistry for epithelial cell proliferation. Gastric lesion index (LI) was assessed using planimetry. RESULTS: Expression of COX-2 was enhanced mainly in surface epithelial cells and neck cells following HCl-administration. At 24 h following acid administration,PCNA labeling index (PCNA-LI) was (22.72?4.33) % and (21.98?5.18) % in the groups treated with 40 mg/kg of NS-398 and indomethacin respectively,which was significantly lower than that in the control group [ (34.46?3.61) %,P
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AIM: To clarify the effects of gastrin on t he expression of cyclooxygenase (COX) and several growth factors in rat gastric mu cosa. METHODS: Male Sprague Dawley rats were fasted for 24 hours and s ubcutaneously injected with saline or gastrin 17 at doses of 1 ?g/kg, 10 ?g/kg and 100 ?g/kg, respectively. The expression of COX-1, COX-2, heparin-binding e p idermal growth factor-like growth factor (HB-EGF) and hepatocyte growth factor ( HGF) in the gastric mucosa were examined using Western blotting and immunohistoc hemical staining. Effects of a potent gastrin receptor antagonist YM022 on the e xpression of COX-1, COX-2, HB-EGF and HGF in gastric mucosa were also evaluated. RESULTS: Gastrin dose-dependently increased the expression of C OX-2 and HB-EGF in rat gastric mucosa while the expression of COX-1 and HGF did not change significantly after treatment with gastrin. However, pretreatment wit h YM022 dose-dependently abolished the up-regulation of COX-2 and HB-EGF express ion induced by gastrin. CONCLUSIONS: This study demonstrates that gastrin up-regulates C OX-2 and HB-EGF expression in rat gastric mucosa, indicating that COX-2 and HB-E GF are involved in pathogenesis of the gastrin-related gastric mucosal hyperplas ia and carcinoma of stomach.