Objective:To compare the difference between the combined diagnostic effect of plasma Septin9(SEPT9) and polyligand Syndecan-2(SDC2) methylation with four serum tumor markers in the diagnosis of colorectal cancer.Methods:In this study, 128 patients who were treated in the affiliated Hospital of Xuzhou Medical University from March to December in 2019 were selected for a case-control study.All the subjects were examined by gastroenteroscopy.According to the pathological results, they were divided into three groups: colorectal cancer group( n=74) and colorectal adenoma group( n=7). The patients with no abnormal or inflammatory polyps or proliferative polyps examined by gastroenteroscopy were taken as the control group( n=47). The methylation levels of SEPT9 gene and SDC2 gene were detected by Roche Lightcycler 480 II real-time fluorescence quantitative polymerase chain reaction, and the concentrations of alpha-fetoprotein, carcinoembryonic antigen, carbohydrate antigen 125 and carbohydrate antigen 199 were detected by Roche Cobas 8000 electrochemiluminescence instrument.Chi-square test was used to compare the positive rate of each marker in the three groups.Medcalc was used to draw the receiver operating characteristic curve (ROC) curve of the subjects′ working characteristic curve, and the value of each index in the diagnosis of colorectal cancer was analyzed. Results:The positive rates of SEPT9 gene and SDC2 gene methylation were 81.1%(60/74) and 67.6%(50/74) respectively in colorectal cancer group, and increased to 85.1%(63/74) after combined detection.The positive detection rates of alpha-fetoprotein, carcinoembryonic antigen, carbohydrate antigen 125 and carbohydrate antigen 199 in colorectal cancer group were 1.4%(1/74), 33.8%(25/74), 6.8%(5/74) and 13.5%(10/74), respectively.When the four tumor markers were detected together, the positive detection rates were only increased to 43.2%(32/74), except for AFP and carbohydrate antigen 125(χ 2=3.847, 2.430, all P>0.05). The differences were statistically significant (χ 2=48.230, 30.487, 43.285, 3.847, 8.788, 6.988, 8.722, all P<0.05). The area under the curve (AUC) of SEPT9 methylation, SDC2 methylation, alpha fetoprotein, carcinoembryonic antigen, carbohydrate antigen 125 and carbohydrate antigen 199 were 0.854 (0.781, 0.910), 0.795 (0.715, 0.861), 0.575 (0.485, 0.662), 0.685 (0.597, 0.764), 0.603 (0.513, 0.689) and 0.631 (0.541, 0.715), respectively.The AUC of combined detection of two DNA methylation markers was better than that of alpha fetoprotein, carcinoembryonic antigen, carbohydrate antigen 125 and carbohydrate antigen 199, and the differences were statistically significant (alpha fetoprotein: Z=4.990, P<0.001; carcinoembryonic antigen: Z=3.743, P<0.001; carbohydrate antigen 125: Z=4.951, P<0.001; carbohydrate antigen 199: Z=3.983, P<0.001). The combined detection of two kinds of gene methylation was better than the combined detection of four kinds of serum markers in the diagnosis of colorectal cancer, and the difference was statistically significant ( Z=3.334, P<0.001). Conclusion:The combined detection of SEPT9 gene and SDC2 gene methylation in plasma is more suitable for non-invasive diagnosis of colorectal cancer than the combined detection of 4 serum tumor markers.

Objective:To explore the clinical characteristics and risk factors of incomplete endoscopy of small intestine capsule.Methods:From August 2015 to February 2019, the data of 282 patients with small intestine examination by capsule endoscopy in Affiliated Hospital of Xuzhou Medical University were analyzed retrospectively.The clinical and laboratory examination data were statistically analyzed.The risk factors of incomplete small intestine examination were analyzed by multivariate logistic regression.Results:In all 282 patients, 45 cases did not complete the examination, the incomplete rate was 16.0% (45/282). Multivariate analysis showed that male ( OR=3.261, 95% CI1.084-9.815, P=0.035), age (64.78±13.23) years ( OR=1.046, 95% CI1.007-1.086, P=0.020), intragastric time (59.78 (27.89, 108.67)) min ( OR=1.014, 95% CI1.005-1.024, P=0.004), gastrointestinal bleeding ( OR=3.017, 95% CI1.025-8.880, P=0.045), abdominal operation history ( OR=4.902, 95% CI1.553-15.470, P=0.007) were risk factors for incomplete examination.Hemoglobin (95.20±26.16) g/L( OR=0.977, 95% CI0.957-0.998, P=0.033), albumin (35.73±7.11) g/L( OR=0.871, 95% CI0.790-0.961, P=0.006) were protective factors for incomplete examination. Conclusion:Male, age, intragastric time, gastrointestinal hemorrhage and abdominal operation history were risk factors for incomplete small bowel examination with capsule endoscopy, and hemoglobin and albumin were protective factors.Before capsule endoscopy, patients should not only evaluate the intestinal stenosis, but also pay attention to the nutritional status and clinical symptoms of patients, so as to improve the completion rate of patients.

Objective To investigate the relationship between Helicobacter pylori infection and serum gastrin 17 levels and colorectal polyps. Methods The clinical data of 214 patients with colorectal polyps who underwent gastrointestinal endoscopy at the Affiliated Hospital of Xuzhou Medical University from June 2017 to April 2019 were collected. The specimens were divided into two groups after pathological diagnosis. The group included 126 cases with adenomatous polyps (adenomatous polyps group) and 88 cases with hyperplastic polyps( hyperplastic polyps group) . Another 89 patients without obvious abnormality were selected as the control group. Serum Hp antibody was detected by western blot,and serum gastrin 17 levels were quantitatively detected by ELISA. Hp infection rate and serum gastrin 17 levels were compared between the groups. Results The infection rate of 74. 2%( 66 ／ 89) type I HP in adenomatous polyp group was significantly higher than that of 55. 6%( 30 ／ 54) in proliferative polyp group and 48. 7%( 19 ／ 39) in control group,the difference was statistically significant( χ2＝5. 271,P＝0. 022; χ2＝7. 867,P＝0. 005). The infection rate of type I HP in proliferative polyp group was not statistically significant ( P＞0. 05) . The HP infection rate in colorectal polyp group was 66. 8%(143 ／ 214) and 67. 1%(96 ／ 143) respectively,which was significantly higher than that in control group (43. 8%(39 ／ 89) and 48. 7%(19 ／ 39),the difference was statistically significant ( χ2 ＝ 13. 87, 4. 467, all P＜0. 05 ) . The infection rate of Hp in colorectal proliferative polyp group was 61. 4%(54／88) and adenomatous polyp group was 70. 6%(89／126) higher than that in control group (43. 8%(39／89),the difference was statistically significant( χ2＝5. 46,15. 57,all P＜0. 05) . The serum gastrin 17 level in adenomatous polyp group (11. 35 ( 6. 67,20. 87) pmol／L) was significantly higher than that in proliferative polyp group (7. 88(3. 11,13. 07) pmol／L) and control group (5. 69 (2. 94,11. 37) pmol／L), the difference was statistically significant ( Z＝4. 91, all P＜0. 05) . The serum gastrin 17 level in adenomatous polyps group with type I Hp infection (14. 35 (8. 12,23. 68) pmol／L) was significantly higher than that of Hp-negative patients ( 8. 42 ( 2. 42, 20. 84) pmol／L), and the difference was statistically significant (Z＝2. 87,P＜0. 05). Conclusion HP infection is closely related to colorectal polyps,especially adenomatous polyps. The increased level of serum gastrin 17 is a risk factor for colorectal adenomatous polyps. Type I HP infection can increase the level of serum gastrin 17,and it is more closely related to adenomatous polyps.

Objective: To investigate the effects of glutamate (Glu) injected into hypothalamic paraventricular nucleus (PVN) on visceral pain of chronic visceral hypersensitivity (CVH) rats and its possible mechanism. Methods: Newborn SD rats were given CVH rat model by colorectal distension (CRD) on the 8th, 10th and 12th day after birth. Thirty rats with successful CVH model were randomly divided into CVH model group (CVH group), CVH + injection of saline into PVN group (NS group), CVH+ injection of Glu into PVN (3, 6, and 12 μg Glu, namely G3, G6, and G12, respectively), 6 rats in each group, and 6 SD rats with matching body mass were taken as sham operation group (Sham group). The pain behavior of the rats was evaluated by pain threshold, abdominal withdrawal reflex (AWR) score, and abdominal external oblique muscle electromyography (EMG). The expression of arginine vasopressin (AVP) and the proliferation of colon tissue were detected by immunohistochemical staining. The apoptosis of colon tissue was detected by TUNEL. Results: Compared with the NS group, the pain thresholds of the G3, G6 and G12 groups increased, and the AWR scores and EMG amplitudes decreased. The differences were statistically significant(Pain threshold: t=7.65, 16.31, 24.78, both P<0.05; AWR scores: t=-2.98, -4.77, -7.29, both P<0.05; EMG amplitudes: t=-3.06, -5.75, -8.92, both P<0.05). Compared with the Sham group, the expression of AVP in PVN of the CVH group and NS group decreased ((42.63±5.20) %, (18.67±2.94) %, (17.53±2.47) %; t=6.95, t=7.56, both P<0.05). The expression of AVP was increased after different doses of Glu into PVN, and the AVP level in G12 group ((18.15±6.49)%) was higher than that of NS group, the difference was statistically significant (t=-4.21, P<0.05). Compared with the Sham group, the expression of PCNA in colonic mucosal cells of the CVH group and NS group decreased ((65.48±1.68) %, (18.39±1.67) %, (17.69±1.68) %; t=34.35, t=34.80, both P<0.05). The expression of PCNA was increased after different doses of Glu injected into PVN, and the PCNA level in G12 group ((59.91±5.63)%) was higher than that of NS group, the difference was statistically significant (t=-12.44, P<0.05). Compared with the Sham group, the expression of apoptotic cells in colonic mucosal cells of the CVH group and NS group increased ((23.38±11.40)%, (83.79±3.57)%, (80.91±2.47)%; t=-8.77, t=-8.54, both P<0.05). The expression of apoptotic cells was decreased after different doses of Glu into PVN, and the G12 group was ((18.15±6.49) %). Compared with NS group, the difference was statistically significant (t=15.65, P<0.05). Conclusion Injection of Glu into hypothalamic PVN can alleviate the visceral pain behaviors in CVH rats, and its mechanism may be related to arginine vasopressin.

Objective To analyze the serum gastric function and Helicobacter pylori ( HP ) infection in patients with gastric hyperplastic polyps and gastric fundic gland polyps.Methods From December 2017 to December 2018, 135 patients with gastric polyps and pathologically confirmed gastric hyperplastic polyps and gastric fundic gland polyps were enrolled in the hospital of Xuzhou Medical University.Among them, 68 patients with hyperplastic polyps, 67 cases of the gastric fundic gland polyps.Serum Hp antibodies ( UreA, UreB, VacA, CagA antibodies ) were qualitatively detected by immunoblotting.Eighty patients with chronic superficial gastritis were selected as the control group.Three groups of serum pepsinogen?I ( PG?I),pepsinogen?Ⅱ( PG?Ⅱ),gastrin were detected by enzyme?linked immunosorbent assay (ELISA).Gastrin?17( G?17) and calculate PGⅠ and PGⅡ ratio( PGR).Results The levels of serum PGⅡ(13.13(8.15,20.30) μg ／L) and G17 (8.44(3.72,27.17) pmol／L) in the gastric hyperplastic polyp group were higher than those in the control group (9.16(5.56,15.14) μg／L and 1.83(0.87,5.95) pmol／L) ( P＜0.05),and the PGR level was lower than the control group ( P＜0.05);serum PGI ( 120.12 ( 86.72,174.70) μg／L), PGII ( 11.92 ( 7.27,22.26) μg／L),G17 ( 5.68 ( 1.79, 14.65) pmol／L) in the gastric fundic gland polyp group was higher than the control group (( 101.32 (79.17,131.33) μg ／L,9.16 ( 5.56,15.14) μg ／L,1.83 ( 0.87,5.95) pmol／L) ( P 均＜0.05)) ( P＜0.05); serum G17 (8.44(3.72,27.17) pmol／L) level in gastric hyperplastic polyp group was higher than gastric fundus polyp group (5.68(1.79,14.65) pmol／L) ( P＜0.05); Hp infection rate in gastric hyperplastic polyp group61.76%(42／68)was higher than that in the gastric fundic gland polyp group40.30%(27／67) (P＜0.05),and type I Hp was the main one (P＜0.05).The serum PGⅡ and G17 levels in the gastric hyperplastic polyp group were higher than those of Hp negative ( all P＜0.05).There were no significant differences in serum PGI, PGⅡ, G17, and PGR levels between the HP?positive and negative?positive patients in the gastric fundus polyp group.The serum PGI and PGR levels in the hypertrophic polyp group were higher than those in the HPⅡ type ( all P＜0.05).There was no significant difference in the levels of serum PGⅠ,PGⅡ,G17,and PGR between the gastric fundic gland polyps group and the type Ⅱ.Conclusion Serum PG and G17 levels in patients with gastric hypertrophic polyps and gastric fundic gland polyps are higher than those in patients with chronic superficial gastritis.Patients with gastric hyperplastic polyps have higher HP infection rate and abnormal gastric function than gastric fundic gland polyps.

Background:Cerebellar fastigial nucleus (FN)is involved in regulation of visceral activities such as cardiovascular, ingestion,respiratory,and acute gastric mucosal injury,yet it is unclear whether it participates in the regulation of visceral hypersensitivity and what is the possible mechanism. Aims:To investigate the effect and possible mechanism of glutamic acid (Glu ) injection into cerebellar FN on chronic visceral hypersensitivity in rats. Methods: Chronic visceral hypersensitivity rat model was established by neonatal colorectal distension (CRD). After 8 weeks,the rats were divided into CRD group,solvent group (0. 2 μL 0. 9% NaCl solution injection into cerebellar FN),high-,medium-,low-dose Glu groups (12,6,3 μg Glu injection into cerebellar FN,respectively),3-MPA +Glu group (12 μg Glu injection after glutamate decarboxylase inhibitor 3-MPA injection into cerebellar FN),Bic + Glu group (12 μg Glu injection into cerebellar FN after GABAAreceptor blocker Bic injection into lateral hypothalamic area). Pain threshold,abdominal withdrawal reflex (AWR)score and abdominal external oblique muscle electromyography (EMG)were used to detect visceral sensitivity,and malondialdehyde (MDA)content and superoxide dismutase (SOD)activity were measured. Results:Chronic visceral hypersensitivity rat model was successfully established. Compared with CRD group,pain threshold was significantly increased (P＜0. 05),AWR score,EMG amplitude,MDA content were significantly decreased (P＜0. 05 ),and SOD activity was significantly increased in a dose-dependent manner in Glu group (P ＜0. 05 ). Compared with 12 μg Glu group,pain threshold was significantly decreased (P＜0. 05),AWR score,EMG amplitude, MDA content were significantly increased (P ＜0. 05),and SOD activity was significantly decreased in 3-MPA +Glu group,Bic+Glu group (P＜0. 05). Conclusions:Glu injection into cerebellar FN can significantly reduce the visceral sensitivity in rats. The mechanism may be that Glu in cerebellar FN produces GABA via glutamate decarboxylase,and then binding GABAAreceptor in lateral hypothalamic area,resulting in increased intestinal mucosal antioxidant capacity, thereby reducing visceral hypersensitivity.

Objective To investigate the effects of NANOG/deleted in breast cancer 1 (DBC1)pathway on biological behavior of gastric cancer cells.Methods From May 2014 to May 2015,25 patients who underwent gastric cancer resection were selected.The expression of NANOG and DBC1 was detected by real time reverse transcription polymerase chain reaction (RT-PCR) and Western blotting in N-tera,SGC-7901,HGC-27,MKN-45,MGC803,NCI-N87,BGC823 cell lines,normal gastric epithelial cell line GES-1 and gastric cancer tissues.The proliferation,apoptosis and colony formation ability of MKN-45 cells in short hairpin (sh)NANOG-1,shNANOG-2,sh-control and shDBC1 groups were determined by MTT assay,flow cytometry and colony forming assay.The effects on the expression of the two genes in MKN-45 cells were verified by polymerase chain reaction (PCR) in shNANOG,sh-control,shDBC1 and shDBC1+NANOG groups and the effects of down regulation of DBC1 on cell biological behavior were further investigated.The differences in gene expression profile after interference which were screened by gene chips and bioinformatics were analyzed.The mechanism of NANOG regulating DBC1 was explored by Dual-luciferase assay.T test was used for two groups comparison while one-way analysis of variance was for multiple groups.Results NANOG and NANOG mRNA were highly expressed in N-tera cells,which were 1.02±0.08 and 0.95 ±0.03,respectively,and the expressions in SGC-7901,HGC-27,MKN-45 and NCI-N87 cell lines were 0.67±0.03 and 0.64±0.04,0.58±0.02 and 0.28±0.02,0.83±0.03 and 1.04 ± 0.05,and 0.61 ± 0.02 and 0.64 ± 0.08,respectively;no expression was detected in normal gastric epithelial cell line GES-1,and the expressions in MKN-45 cells were the highest in gastric cancer cells (F=21.51 and 85.53,both P＜0.01).The expression of DBC1 in HGC-27,MGC803,NCIN87,SGC-7901,BGC823 and MKN-45 cells were 0.37±0.02,0.33±0.02,0.42±0.01,0.58±0.04,0.33±0.05,and 0.87±0.02,respectively;while there was no expression of NANOG,NANOG mRNA and DBC1 in GES-1 cells.The expression of NANOG mRNA and DBC1 was detected in gastric cancer tissues of 24.0％ (6/25) patients.Compared with that of the sh control group,the apoptosis rates of MKN-45 cells in the shNANOG-1,shNANOG-2 groups were increased ((2.24±0.17)％ vs (6.03±0.24) ％ and (6.95 ± 0.38) ％),and the difference was statistically significant (F =81.18,P ＜ 0.01).Compared with that of the sh-eontrol group,the colony forming abilities of MKN-45 cells in the shNANOG-1 and shNANOG-2 groups were significantly decreased (172.03±6.35 vs 74.32±5.32 and 53.08±3.82),and the difference was statistically significant(F=171.61,P＜0.01).The results of PCR showed that compared with that of sh-eontrol group,the expression levels of NANOG mRNA and DBC1 mRNA in shNANOG group were lower (1.04±0.05 vs 0.54±0.03,1.08±0.08 vs 0.42±0.03),the level of DBC1 mRNA in shDBC1 group was lower (1.08±0.08 vs 0.50±0.04),and the differences were statistically significant (t=9.15,7.37 and 6.06,all P＜0.01).The expression level of NANOG mRNA in shDBC1 + NANOG group was higher (1.04 ± 0.05 vs 3.01 ± 0.08),while the expression level of DBC1 mRNA was lower (1.08 ± 0.08 vs 0.71 ± 0.06),and the difference was statistically significant (t=-20.22 and 3.74,both P＜0.05).The expression level of DBC1 mRNA in shDBC1±NANOG group was higher than that in shDBC1 group (0.71±0.06 vs 0.50±0.04),and the difference was statistically significant (t=4.00,P＜0.05).Bioinformatic analysis showed that DBC1 gene promoter region had the potential NANOG protein binding sites.Dual-luciferase assay indicated NANOG played the role in transcription activation in DBC1 promoter regions.Conclusion NANOG and DBC1 are highly expressed in various gastric cancer cell lines.NANOG may affect the proliferation,apoptosis and colony formation of MKN-45 cells by regulating the expression of DBC1.NANOG/DBC1 pathway may be a promising new target of gastric cancer treatment.

Objective:To explore the mechanism of the cytotoxicity of human NK cells induced by atorvastatin to colon cancer cell lines. Methods:After colon cancer cells (HCT-116,SW-480,Caco-2) were cultured with different concentrations of atorvastatin, CCK-8 assay was used to assess the effect of atorvastatin on growth of colon cancer cells. The amplification of human NK cells was induced by SCGM medium in vitro. Automatic biochemical analyzer was applied to test the cytotoxicity of NK cells to colon cancer cells which cultured with different concentration of atorvastatin. FCM was used to detect the expression rate of MICA/B on the cells. Results:(1) The cultivation of NK cells:The proportion of NK cells attained to 93. 1% from 4. 5% after cultured for 10 days. (2) The effects of atorvastatin on the growth of the colon cancer cells:After cultured with atorvastatin,the inhibition rate of HCT-116 cells was higher than that in control when the density of atorvastatin increased from 5 μmol/L to 40 μmol/L after 48 h and from 1. 25 μmol/L to 40 μmol/L after 96 h ( P<0. 05 ) . Correlation analysis showed that the concentration of atorvastatin and the growth inhibition rate of HCT-116 cells were positively correlated(r[48 h]=0. 13,r[96 h]=0. 22,P<0. 05). (3) The cytotoxicity of NK cells to colon cancer cells effected after atorvastatin: In different atorvastatin concentrations groups,the cytotoxicity of NK cells to three colon cancer cell lines was all higher than that in control ( P<0. 05 ) . The atorvastatin concentration was from 2. 5 μmol/L to 10 μmol/L for HCT-116 cells,from 5 μmol/L to 20μmol/L for SW-480 cells,and from 2. 5μmol/L to 20μmol/L for Caco-2 cells. Among the three cell lines, the cytotoxicity of NK cells to HCT116 was the highest in the same concentration. (4)NK cells by atorvastatin cutting statins 96 h,the concentration of 20 mmol/L and 40 mmol/L inhibition rate was higher than that of control group,more than other groups on NK cell growth without significant effect. ( 5 ) The impact of atorvastatin on MICA/B expression of colon cancer cells: After cultured with different concentrations of atorvastatin,the expression of MICA/B on colon cancer cells was higher than that in control(P<0. 05). The concentration was 2. 5μmol/L and 5μmol/L for HCT-116 cells,10μmol/L and 20μmol/L for SW-480 cells,and from 2. 5μmol/L to 40 μmol/L for Caco-2 cells. Conclusion:Atorvastatin could inhibit the growth of colon cancer cells (HCT-116,SW-480 and Caco-2) in a dose-dependent manner;and it could enhance the cytotoxicity of NK cells to colon cancer cells;it also could promote the expression of MICA/B of colon cancer cells,and improve the immunogenicity of colon cancer cells.

Objective To evaluate the clinical effectiveness treatment of chronic gastritis by Kangfuxin liquid.Methods Computer -based online was used to retrieve Cochrane library,PubMed,CNKI,Wanfang data,VIP database(since build library retrieval time),to find the Kangfuxin liquid and add and subtract randomized controlled trials for the treatment of CAG Meta analysis was performed to evaluate the data by using RevMan5.3 software.Results Six articles were included in the study,a total of 742 patients.Meta analysis results showed that the total effective rate of Kangfuxin liquid in the treatment of chronic atrophic gastritis was better than that of routine drugs (OR =6.00,95 ％ CI:3.65-9.86,P ＜ 0.05).The helicobacter pylori eradication rate of Kangfuxin liquid in the treatment of chronic atrophic gastritis was better than that of routine drugs(OR =3.71,95％ CI:1.89-7.29,P ＜ 0.05).The effect of Kangfuxin liquid together with traditional triple therapy was better than traditional triple therapy (OR =6.15,95 ％ CI:3.24-11.68,P ＜ 0.05).Conclusion The effect of Kangfuxin liquid in the treatment of chronic atrophic gastritis is more outstanding than conventional drug treatment.

Objective To evaluate the effecacy of Bifidobacterium viable triple capsule on anti-Helicobacter pylori (Hp) treatment and its adverse reactions during eradication therapy .Methods T rials on the effect of Bifidobacterium viable triple capsule on Hp eradication were analyzed by Meta-analysis with the software Stata 12 .0 for comprehensive quantitative analysis of its clinical efficacy and adverse reactions during treatment .Results Eight randomly controlled trials (n=114 4) were includ-ed .Pooled Hp eradication rates were 85.91% and 69 .21% for patients with and without Bifidobacterium viable triple capsule respectively ,with the odds ratio (OR) being 3 .07 (95% CI=2 .25~4 .19 ,test for overall effect Z= 7 .08 , P<0 .01);the occurrences of total adverse reactions were 7 .58% and 20.87% respectively ,with the summary OR being 0 .42 (95% CI=0 .29~0 .61 ,test for overall effect Z= 4 .48 , P< 0 .01) .The difference of the two groups was statistically significant . Conclusion Current evidence shows that Bifidobacterium viable triple capsule could increase eradication rate of anti-Hp treat-ment and improve adverse reactions during therapy .