Pseudoileus caused by primary visceral myopathy in a Han Chinese patient with a rare MYH11 mutation: A case report.

BACKGROUND: Chronic intestinal pseudo-obstruction (CIPO) is a syndrome of intestinal motor dysfunction caused by intestinal nerve, muscle, and/or Cajal stromal cell lesions. CIPO is a serious category of gastrointestinal dynamic dysfunction, which can eventually lead to the death of patients with intestinal failure. Due to considerable phenotypic heterogeneity, the estimated incidence of CIPO is 1/476190 and 1/416666 in men and women, respectively. According to the etiology, CIPO can be divided into idiopathic and secondary, of which the latter is the most common, often secondary to tumor, virus infection, connective tissue disease, neurological diseases, and endocrine diseases. Idiopathic CIPO in the intestinal tract is divided into visceral myopathy, neuropathy, and stromal cell lesions according to the location. Surgery is usually not recommended for CIPO, because it often does not benefit patients with CIPO, and postoperative intestinal obstruction is likely to occur, which may even worsen the condition. CASE SUMMARY: Here, we describe the case of a 43-year-old male Han Chinese patient with a 15-year history of recurrent abdominal distention with no clear cause. The results of physical, biochemical, and other relevant examinations showed no clear abnormalities. Contrast-enhanced computed tomography (CT) indicated a large duodenum, clear expansion of the intestinal lumen, and CIPO. Whole exome sequencing (WES) of the patient and his mother confirmed the diagnosis of primary familial visceral myopathy type 2 chronic pseudoileus with a rare heterozygous gene mutation in MYH11. This is the second reported case of CIPO with a heterozygous MYH11 [NM_001040113.1: c.5819delC (p.Pro1940Hisfs*91)] mutation. CONCLUSION: This case report indicates that physicians can perform routine clinical examinations, CT, and WES to achieve a diagnosis and treatment of CIPO in early disease stages.

Proton pump inhibitor in the prevention of upper gastrointestinal mucosal injury associated with dual antiplatelet therapy after coronary artery bypass grafting (DACAB-GI-2): study protocol for a randomized controlled trial.

BACKGROUND: Dual antiplatelet therapy (DAPT) is recommended in secondary prevention after coronary artery bypass grafting (CABG), but it is inevitably associated with the risk of bleeding, of which gastrointestinal bleeding accounts for more than half. Proton pump inhibitors (PPIs) may increase the risk of major cardiovascular adverse events when reducing the risk of upper gastrointestinal bleeding. Therefore, the optimal duration of a PPI in combination with DAPT is unclear. METHODS: The "Proton Pump Inhibitor Preventing Upper Gastrointestinal Injury in Patients on Dual Antiplatelet Therapy after CABG" (DACAB-GI-2) study is a prospective, single-center, open-label, parallel, randomized controlled trial. A total of 232 eligible subjects who are scheduled or initiated on DAPT (clopidogrel plus aspirin or ticagrelor plus aspirin) for 12 months immediately after CABG will be enrolled and be randomized in a 1:1 ratio to either a 12-month pantoprazole treatment arm or a 1-month treatment arm. The primary outcome is to assess the rate of gastroduodenal erosions and ulcers evaluated by esophagogastroduodenoscopy (EGD) within 12 months after randomization, based on the modified Lanza score. Secondary outcomes include reflux esophagitis and upper gastrointestinal bleeding. Other pre-specified outcomes include major adverse cardiovascular events, graft failure, and all-cause death. DISCUSSION: This study aims to compare the efficacy and safety of 12 months and 1 month of pantoprazole treatment in preventing DAPT-related upper gastrointestinal mucosal injury after CABG. TRIAL REGISTRATION: ClinicalTrials.gov NCT03908593 .

Upper gastrointestinal mucosal injury associated with ticagrelor plus aspirin, ticagrelor alone, or aspirin alone at 1-year after coronary artery bypass grafting.

BACKGROUND AND AIM: The presence and severity of upper gastrointestinal mucosal lesions have not been evaluated using esophagogastroduodenoscopy (EGD) in patients receiving ticagrelor plus aspirin or alone after myocardial revascularization. We assessed upper gastrointestinal mucosal injury and the use of proton pump inhibitors (PPIs) in patients receiving 1 year of antiplatelet therapy after coronary artery bypass grafting (CABG). METHODS: In this single-center prospective substudy of a randomized trial, 231 patients completing 1-year antiplatelet therapy (ticagrelor 90 mg twice daily plus aspirin 100 mg once daily, ticagrelor 90 mg twice daily, or aspirin 100 mg once daily, in 81, 80, and 70 patients, respectively) after CABG underwent 13 C urea breath testing and EGD. Gastroduodenal lesions were assessed by modified Lanza score, and reflux esophagitis was evaluated according to Los Angeles classification. Additionally, at least one ulcer ≥ 5 mm was separately analyzed. RESULTS: Among 231 patients, EGD showed 28 (12.1%) with ulcers ≥ 5 mm, which were detected in 13.6% (11/81) of ticagrelor plus aspirin recipients, 8.8% (7/80) of ticagrelor recipients, and 14.3% (10/70) of aspirin recipients, and 24 (10.4%) had reflux esophagitis. Eighty-eight (38.1%) patients had a positive 13 C urea breath testing after 1 year of treatment, and one patient received eradication therapy during follow up. Nineteen (8.2%) patients received a PPI for ≥ 6 months. CONCLUSIONS: Severe upper gastrointestinal mucosal lesions were more frequently observed in patients treated with ticagrelor plus aspirin and aspirin monotherapy than in patients treated with ticagrelor monotherapy for 1 year post-CABG. Prophylactic use of PPIs might be inadequate.

Development and Preliminary Evaluation of Chinese Preschoolers' Caregivers' Feeding Behavior Scale.

BACKGROUND: Caregivers' feeding behavior plays a crucial role in the development of overweight and obesity in preschoolers. However, to date, there is no broadly accepted scale or questionnaire for assessing preschoolers' caregivers' feeding behavior in China. OBJECTIVE: To develop a scale that can be used to assess preschoolers' caregivers' feeding behavior in China and to conduct a preliminary evaluation of the scale's reliability, validity, and discriminative ability. DESIGN: The scale was created through a literature review and qualitative interviews with the target population. Items were reviewed by 50 caregivers of preschoolers and 10 experienced pediatricians, and 95 items were selected to form a draft scale. The draft scale underwent three rounds of investigation, and the results from these evaluations were used to select items that formed the final scale. PARTICIPANTS/SETTING: Three groups of caregivers (n=175, 400, and 912) were sampled and stratified from urban and suburban kindergartens in the cities of Jinan and Xi'an between March 2016 and October 2017 to participate in evaluations of the draft scale. From these caregiver groups, 146, 362, and 768 participants completed valid questionnaires, respectively, which were used in the scale's evaluation. PRIMARY OUTCOME MEASURES: The general demographic data of the participants and scores of each item in the scale were the primary outcome measures. STATISTICAL ANALYSES PERFORMED: Exploratory factor analysis and variability analysis were used to evaluate the draft scale, based on data from two rounds of investigation. The structure of the scale was explored through confirmatory factor analysis, and its reliability, construct validity, and discriminative ability were evaluated based on data from a third round of investigation. RESULTS: The Chinese Preschoolers' Caregivers' Feeding Behavior Scale (CPCFBS) consisted of 35 items and seven dimensions; the total cumulative variance contribution rate was 58.6%; the Cronbach's α coefficient was .91; the split-half reliability coefficient was 0.89; and the test-retest reliability coefficient was 0.85. The age and weight status of the children and the caregivers' age and education levels, as well as family incomes and child-caregiver relationships, were correlated with feeding behavior. CONCLUSIONS: The CPCFBS appeared to have good reliability and construct validity in specific Chinese populations. Future studies are needed to confirm existing findings in different Chinese populations with larger sample sizes.

Prevalence, Awareness and Associated Risk Factors of Diabetes among Adults in Xi'an, China.

The study aimed to investigate the prevalence, awareness, treatment and glycaemic control of diabetes and its associated risk factors among adults in Xi'an, China. We collected data among participants aged 18 years or older through a self-developed questionnaire and an additional health examination. A total of 8150 participants were included, with an overall prevalence of diabetes of 8.0%. Among 655 participants with diabetes, 52.5% were aware they had diabetes, 48.1% took antidiabetic treatment, and 19.1% had their fasting blood glucose level at less than 7.0 mmol/l. Older age, lower educational level, higher body mass index, larger waist circumference, having an unhealthy diet and having more comorbidities were positively associated with the risk of diabetes. Participants who were older, who had higher education and who had more comorbidities were more aware that they had diabetes. Being older age, having higher education and having more comorbidities were also factors for better treatment. Participants who were older were more likely to have their glucose level controlled. The prevalence of diabetes among adults in Xi'an is high, with suboptimal awareness, treatment, and glycaemic control rates. Comprehensive integrated strategies based on risk factors should be implemented to improve the prevention and glycaemic control of diabetes.

Prevalence, awareness, treatment, and control of hypertension and associated risk factors among adults in Xi'an, China: A cross-sectional study.

To investigate the prevalence, awareness, treatment, and control of hypertension, and the associated risk factors among adults in Xi'an, China.From October to December 2013, participants in Xi'an, China were recruited for the study by using a multiple-stage sampling method. A self-developed questionnaire with an additional health examination was used to collect data on the history of hypertension diagnosis and antihypertensive medication. The status on prevalence, awareness, treatment, and control of hypertension were analyzed and related risk factors were identified by using logistic regression analysis.A total of 8193 participants were included with an overall prevalence of hypertension of 20.4%. Among the hypertensive participants, 63.7% were aware of their conditions, 47.3% took antihypertensive medication, and 17.8% had their blood pressure (BP) controlled within 140/90 mm Hg. More complications and less frequent BP measurements were associated with hypertension. Older participants, non-drinkers, and those with more complications and more frequent BP measurements were more aware of their hypertension. Being older, living in an urban area, and having more frequent BP measurements were all factors for better treatment. Participants who were women, living in an urban area, with a higher educational level and who were not obese were more likely to have their hypertension controlled.The prevalence of hypertension among adults in Xi'an is high with suboptimal low awareness, treatment, and control rates. Further comprehensive integrated strategies based on these risk factors should be taken into account in order to improve the prevention, awareness, treatment, and control of hypertension.

Gallbladder gallstone disease is associated with newly diagnosed coronary artery atherosclerotic disease: a cross-sectional study.

BACKGROUND AND AIMS: The association between gallstone disease and coronary artery atherosclerotic disease (CAD) remains unclear. To clarify their relationship, patients with CAD newly diagnosed by coronary angiography were investigated in this cross-sectional study. METHODS: The study cohort consisted of 1,270 patients undergoing coronary angiography for the first time between January 2007 and September 2011. Patients with ≥50% diameter stenosis in any major coronary artery on coronary angiography were defined as being CAD positive (n = 766) and those with no stenosis as CAD negative (n = 504). Multivariate logistic regression was used to investigate the relationship between gallstone disease and CAD. The odds ratios (OR) of factors associated with CAD were calculated. In addition, CAD-positive and CAD-negative patients were matched one-to-one by age, gender and metabolic syndrome (MetS), and the association between gallbladder disease and CAD was determined. RESULTS: The prevalence of gallstone disease was significantly higher in CAD-positive than in CAD negative patients (149/766 [19.5%] vs 57/504 [11.3%], P<0.01). Gallstone disease was significantly associated with CAD (adjusted OR = 1.59, 95% confidence interval [CI] 1.10-2.31). Following matched pairing of 320 patients per group, gallstone disease remained significantly associated with CAD (adjusted OR = 1.69, 95% CI: 1.08-2.65). CONCLUSION: Gallstone disease is strongly associated with CAD diagnosed by coronary angiography.

Fatty liver index correlates with non-alcoholic fatty liver disease, but not with newly diagnosed coronary artery atherosclerotic disease in Chinese patients.

BACKGROUND: Fatty liver index (FLI) was recently established to predict non-alcoholic fatty liver disease (NAFLD) in general population, which is known to be associated with coronary artery atherosclerotic disease (CAD).This study aims to investigate whether FLI correlates with NAFLD and with newly diagnosed CAD in a special Chinese population who underwent coronary angiography. METHODS: Patients with CAD (n = 231) and without CAD (n = 482) as confirmed by coronary angiography were included. Among them, 574 patients underwent B-ultrosonography were divided into NAFLD group (n = 209) and non-NAFLD group (n = 365). Correlation between FLI and NAFLD was analyzed using pearson's correlation. The associations between FLI and NAFLD as well as CAD were assessed using logistic regression. The predictive accuracy of FLI for NAFLD was evaluated using receiver operating characteristics (ROC) curve analysis. RESULTS: FLI was significantly higher in NAFLD group (37.10 ± 1.95) than in non-NAFLD group (17.70 ± 1.04), P < 0.01. FLI correlated with NAFLD (r = 0.372, P < 0.001). The algorithm for FLI had a ROC-AUC of 0.721 (95% CI: 0.678-0.764) in the prediction of NAFLD. Logistic regression analysis showed that FLI was associated with NAFLD (adjusted OR = 1.038, 95% CI: 1.029-1.047, P < 0.01). The proportion of patients with CAD did not differ among the groups of FLI ≤ 30 (32.3%), 30-60 (31.0%), and ≥60 (35.3%). No significant association was found between FLI and CAD (adjusted OR = 0.992, 95% CI: 0.981-1.003 in men and OR = 0.987, 95% CI: 0.963-1.012 in women, P > 0.05). CONCLUSIONS: FLI showed good correlation with NAFLD in patients who underwent coronary angiography, but not with newly diagnosed CAD. This might be underestimated because some patients in non-CAD group may have other underlying cardiovascular diseases.

Prognostic factors and outcome of resected patients with gastrointestinal stromal tumors of small intestine.

We document the clinical behavior of gastrointestinal stromal tumors (GISTs) of the small intestine and identify predictors for long-term disease-free survival (DFS) for small intestine GIST patients. From December 2001 to 2008, 114 consecutive patients with mesenchymal tumors involving the small intestine were enrolled. There were 54 male and 60 female (50.6%) patients. After a median follow-up period of 36 months (ranging from 12 to 96 months), recurrence was noted in 19 patients (16.7%) with a median time of 20 months (ranging from 7 to 50 months). There were 12 patients (10.5%) who died of GISTs with a median time from recurrence to death of 14 months (ranging from 8 to 22 months). Univariate analysis by log-rank test indicated that tumor size and mitotic activity were statistically significant for DFS (P = 0.001 and 0.036, respectively). Tumor size was the only significant predictive factor for DFS according to multivariate analysis (P = 0.006). Small tumor size, indicating low risk, predicted more favorable DFS of small intestine GIST patients who underwent curative resection.

Stat3 is involved in angiotensin II-induced expression of MMP2 in gastric cancer cells.

The expression of matrix metalloproteinase-2 (MMP2) has been linked with tumor invasion, angiogenesis, and metastasis. It has been reported that angiotensin II (Ang II) can induce MMP2 expression in gastric cancer cells. However, the molecular basis for Ang II regulates MMP2 expression in gastric cancer cells remains unclear. The aim of our study is to explore whether angiotensin II could induce MMP2 expression mediated through the Stat signaling pathway and its potential mechanism. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP)-binding assays was employed to determine the DNA-STAT binding activity. MMP2 and VEGF expression was analyzed with real-time PCR and Western blots. To examine the role of Stat3 in angiotensin II-induced MMP2 expression, A JAK-specific inhibitor and AG490 were used. Angiotensin II activated STAT-DNA binding activity in dose-dependent manners in gastric cancer cells. AG490 markedly inhibited angiotensin II-induced Stat3 activation and the expression of MMP2 and VEGF in gastric cancer cells. These results indicate that Stat3 may at least in part mediate angiotensin II-induced MMP2 mRNA expression in human gastric cancer cells. The activation of the JAK/Stat3 signaling pathway plays an important role in the progression of gastric cancer and that blockade of JAK/Stat3 signals may provide a novel therapeutic strategy for gastric cancer.

Suppression of gastric cancer growth by baculovirus vector-mediated transfer of normal epithelial cell specific-1 gene.

AIM: To study the inhibitory effect of baculovirus-mediated normal epithelial cell specific-1 (NES1) gene therapy on gastric cancer (GC) in vitro and in vivo. METHODS: We first constructed recombinant baculovirus vectors and then transfected them into gastric cancer cells (SGC-7901). Efficiency of the baculovirus for gene transfer into SGC-7901 cells and cell growth curves were detected by fluorescence microscopy, Western blot and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in vitro, respectively. The therapeutic effect of this gene therapy on GC was confirmed in xenografted nude mice. Tumor growth was determined by tumor volume, and expression of NES1 in tumor was analyzed by immunohistochemistry. RESULTS: Baculovirus vectors were successfully transfected into SGC-7901 cells. SGC-7901 cells transfected with the NES1 gene inhibited cell growth. In the Bac-NES1 treated group, tumor growth was significantly reduced with a high level of NES1 expression CONCLUSION: Baculovirus-mediated NES1 gene can be used in gene therapy for GC.

Angiotensin II type 1 receptor antagonist suppress angiogenesis and growth of gastric cancer xenografts.

Angiotensin II (Ang II) has been reported to promote tumor progression, tumor growth and angiogenesis in many cancers. We previously observed that angiotensin II type 1 receptors (AT1R) were upregulated in human gastric cancer and may be involved in the progression of gastric cancer. We studied the effects of AT1R antagonist on angiogenesis and growth in gastric cancer xenografts to observe the mechanism action of AT1R in the gastric cancer. The results showed that the growth of gastric cancer cells was significantly suppressed by treatment with AT1R antagonist. In vivo, TCV-116, at doses of both 2 and 5 mg/kg/day, significantly suppressed tumor growth in mice (47.3 and 70.2%). Microvessel density was significantly decreased by TCV-116 (3.4 +/- 0.9 and 2.8 +/- 0.5 per field) compared with the control group (12.9 +/- 1.1 per field), and VEGF expression was significantly suppressed by AT1R antagonist. These results demonstrate that AT1R plays an important role in the progression of gastric cancer. Suppression tumor angiogenesis could be one of the mechanisms by which AT1R antagonist suppresses the growth of gastric cancer. These findings also provide a theoretical basis for the future clinical application of AT1R antagonist against gastric cancer.

Angiotensin II type 1 receptor expression in human gastric cancer and induces MMP2 and MMP9 expression in MKN-28 cells.

Angiotensin II (Ang II), a main effector peptide in the renin-angiotensin system, acts as a growth-promoting and angiogenic factor via angiotensin II receptor1 (AT1R). In this study, we investigated the expression of angiotensin II type1 receptor (AT1R) in gastric cancer and the effects of Ang II on the expression of MMP2 and MMP9 in the human gastric cancer cell line MKN-28 cells. The expression of the Ang II type I receptor was examined by western and immunocytochemistry in gastric cancer cell lines and detected by real-time PCR and immunohistochemistry in normal and gastric cancer tissues. The expression of MMP2 and MMP9 were detected by real-time PCR and western after treatment with Ang II and/or AT1R antagonist. AT1R were expressed in all human gastric cancer lines and the expression of AT1R was significantly higher in cancer tissues than that in normal gastric tissues (P < 0.01). Furthermore, Ang II promoted the expression of MMP2 and MMP9 in MKN-28 cells, and the stimulatory effects of Ang II could be blocked by AT1R antagonist. These results suggest that AT1R is involved in the progression of gastric cancer and may promote the angiogenesis of gastric cancer cell line (MKN-28), and these effects may be associated with the upregulation of MMP2 and MMP9.

Downregulation and CpG island hypermethylation of NES1/hK10 gene in the pathogenesis of human gastric cancer.

The normal epithelial cell-specific-1 (NES1)/Kallikrein 10 gene is proposed to be a novel putative tumor suppressor gene in several malignant diseases. The role of NES1 gene in gastric cancer has not been fully understood. Our study revealed that CpG island hypermethylation plays an important role in the downregulation of NES1 mRNA expression in gastric cancer. In situ hybridization showed that loss or reduction of NES1 mRNA expression is associated with differentiation level during tumor progression suggesting that NES1 inactivation might contribute to the malignant progression of human gastric cancers.

HSF1 down-regulates XAF1 through transcriptional regulation.

Studies have indicated the role of HSF1 (heat-shock transcription factor 1) in repressing the transcription of some nonheat shock genes. XAF1 (XIAP-associated factor 1) was an inhibitor of apoptosis-interacting protein with the effect of antagonizing the cytoprotective role of XIAP. XAF1 expression was lower in gastrointestinal cancers than in normal tissues with the mechanism unclear. Here we showed that gastrointestinal cancer tissues expressed higher levels of HSF1 than matched normal tissues. The expression of XAF1 and HSF1 was negatively correlated in gastrointestinal cancer cell lines. Stress stimuli, including heat, hypo-osmolarity, and H2O2, significantly suppressed the expression of XAF1, whereas the alteration of HSF1 expression negatively correlated with XAF1 expression. We cloned varying lengths of the 5'-flanking region of the XAF1 gene into luciferase reporter vectors, and we evaluated their promoter activities. A transcription silencer was found between the -592- and -1414-nucleotide region that was rich in nGAAn/nT-TCn elements (where n indicates G, A, T, or C). A high affinity and functional HSF1-binding element within the -862/-821-nucleotide region was determined by electrophoretic mobility shift assay and chromatin immunoprecipitation assay. Inactivation of this "heat-shock element" by either site-directed mutation or an HSF1 inhibitor, pifithrin-alpha, restored the promoter activity of the silencer structure. Moreover, pretreatment with antioxidants suppressed HSF1 binding activity and increased the transcriptional activity and expression of XAF1. These findings suggested that endogenous stress pressure in cancer cells sustained the high level expression of HSF1 and subsequently suppressed XAF1 expression, implicating the synergized effect of two anti-apoptotic protein families, HSP and inhibitors of apoptosis, in cytoprotection under stress circumstances.

Activation of STAT3 signaling in human stomach adenocarcinoma drug-resistant cell line and its relationship with expression of vascular endothelial growth factor.

AIM: To investigate the difference in activation of STAT3 signaling between two human stomach adenocarcinoma cell lines: 5-fluorouracil resistant cell line and its parental cell line, and to evaluate its relationship with the expression of vascular endothelial growth factor (VEGF). METHODS: Western blot and electrophoretic mobility shift assay (EMSA) were used to detect the expression of phospho-STAT3 protein and constitutive activation of STAT3 in two human stomach adenocarcinoma cell lines, 5-fluorouracil resistant cell line SGC7901/R and its parental cell line SGC7901, respectively. The mRNA expression of VEGF was analysed by semi-quantitative RT-PCR. The expressive intensity of VEGF protein was measured by immunocytochemistry. RESULTS: The expressions of phospho-STAT3 protein and constitutive activation of STAT3 between two human stomach adenocarcinoma cell lines were different. Compared with the parental cell line SGC7901, the STAT3-DNA binding activity and the expressive intensity of phospho-STAT3 protein were lower in the drug-resistant cell line SGC7901/R. The expression levels of VEGF mRNA and its encoded protein were also decreased in drug-resistant cell line. CONCLUSION: Over-expression of VEGF may be correlated with elevated STAT3 activation in parental cell line. Lower VEGF expression may be correlated with decreased STAT3 activation in resistant cell line, which may have resulted from negative feedback regulation of STAT signaling.

[Constitutive activation and clinical significance of Stat3 in human gastric cancer tissues and cell lines].

OBJECTIVE: To investigate the activation of signal transducers and activators of transcription 3 (Stat3) in different types of gastric cancer cell lines and tissues and evaluate the relationship with their clinicopathological parameters. METHODS: Western blotting and electrophoretic mobility shift assay (EMSA) were used to detected the expression of Stat3 protein and Stat3 DNA-binding activity in normal human gastric epithelial cell line 3T3 and five gastric cancer cell lines with different differentiation: MKN28, SGC7901, MKN45, AGS and NCI-SNU-1, respectively. The localization of phospho-Stat3 was determined by immunocytochemistry. The expressive intensity of phospho-Stat3 protein in 50 cases of gastric cancer tissues and adjacent normal mucosa were measured by immunohistochemistry. RESULTS: Compared with normal gastric epithelial cell line 3T3, elevated activities of Stat3 were found in five different human gastric cancer cell lines. The Stat3 DNA-binding activity in moderately and poorly differentiated stomach adenocarcinoma cell lines (SGC7901, MKN45 and AGS) was higher than that of other cell lines (MKN28 and NCI-SNU-1). Phospho-Stat3 was detected primarily in the nuclei of AGS cells. The expressive intensity of phospho-Stat3 protein was significantly increased in gastric cancer tissues as compared with the adjacent normal gastric mucosa, especially in moderately and poorly differentiated cancers (both P < 0.05). The expressive intensity of phospho-Stat3 protein in stage II and stage III tumors was higher than that in stage I tumors (P < 0.05). No statistic difference of phospho-Stat3 expression was found between stage IV and stage I tumors (P > 0.05). The expression of phospho-Stat3 was closely correlated with the differentiation of gastric cancer. CONCLUSION: Elevated activity of Stat3 can be found in different types of human gastric cancer cell lines and gastric cancer. JAK/STAT signal transduction pathway may play an important role in the development of human gastric cancer.

[The impact of decreased Stat3 activation on 5-fluorouracil resistance of human gastric cancer cell line].

OBJECTIVE: To investigate the relationship between different activation of Stat3 signaling and the drug resistance mechanisms in two human gastric cancer cell lines, 5-fluorouracil (5-FU) resistant cell line and its parental cell line. METHODS: Electrophoretic mobility shift assay and Western blot were used to detected Stat3 DNA-binding activity and the expression of phospho-Stat3 protein in 5-FU resistant cell line SGC7901/R and its parental cell line SGC7901, respectively. The mRNA expression of Stat3 and vascular endothelial growth factor (VEGF) were analysed with semi-quantitative RT-PCR. The expressive intensity of VEGF protein was measured by immunocytochemistry. RESULTS: The constitutive activation of Stat3 and the expression of phospho-Stat3 protein were different in two human gastric cancer cell lines. Compared with the parental cell line SGC7901, the Stat3-DNA binding activity and the expressive intensity of phospho-Stat3 protein were lower in the drug-resistant cell line SGC7901/R. The expression level of Stat3 mRNA was also decreased in drug resistant cell line, so did VEGF mRNA and its encoded protein. CONCLUSIONS: The decreased Stat3 activation in 5-FU resistant human gastric cancer cell line SGC7901/R is related to the drug resistance mechanisms and may be correlated with the lower VEGF expression.

[An improved quantitative method for evaluation of ischemic injury and neuroprotection in mouse brain slices].

OBJECTIVE: To establish a simpler and more accurate method for evaluating in vitro ischemic injury and neuroprotective effects of drugs through improving experimental instrument and quantitative index in mouse brain slices. METHODS: An incubation instrument was developed and its application tested. 2,3,5-triphenyltetrazolium chloride (TTC) was used as a substrate to biosynthesize formazan standard in mouse brain slices, and formazan was isolated, purified and identified. Ischemic injury of mouse brain slices was induced by oxygen/glucose deprivation (OGD), the produced formazan from TTC in the cortex and striatum was measured at 490 nm spectrophotometrically. Edaravone and ONO-1078 were added into the incubation medium to observe their neuroprotective effects. RESULT: The incubation instrument worked well for incubating brain slices and obtaining stable results efficiently. Standard formazan was biosynthesized and purified with a purity of 99.3%, and showed a linear range of 0.05 - 1 mg/ml in absorbance at 490 nm (r=0.9997). OGD decreased formazan production in the cortex and striatum in a duration-dependent manner. Edaravone (0.01 to 1 micromol/L) recovered OGD-induced decrease of formazan production, but ONO-1078 showed no effect. CONCLUSION: The incubation instrument and quantitative measurement of formazan developed in this study are efficient,accurate and simple for evaluating ischemic injury and neuroprotection,which can be used in screening of neuroprotective drugs in vitro.