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Objective:To investigate the expression and correlation of serum trefoil factor 3 (TFF3), serum secreted frizzled-related protein 5 (SFRP5), galectin-3 (Gal-3), and nesfatin-1 (NES-1) in patients with type 2 diabetes(T2DM), diabetic nephropathy(DN), chronic kidney disease (CKD), and healthy controls. To explore the relationship between the above factors and the diagnosis of DN and to establish a diagnostic formula for the diagnosis of DN combined with the above four factors.Methods:In each group 36 patients hospitalized in Tianjin Third Central Hospital from April 2017 to June 2019 were enrolled. 36 healthy volunteers were also chosen as the healthy control group. After 8 to 10 hours of fasting, the venous blood of the subjects in each group was centrifuged, the serum was collected for detection, the serum levels of TFF3, SFRP5, Gal-3, and NES-1 were compared, and the Pearson correlation analysis was performed. According to whether the diagnosis of DN was repeated, the subjects were divided into the DN group and the non-DN group (including a healthy control group, T2DM group, and CKD group). The four datasets were analyzed by binary logistic regression, and the diagnostic prediction model of DN was established, which was further verified by receiver operating characteristic (ROC).Results:The serum levels of TFF3, Gal-3 and NES-1 in DN groups were significantly higher than those in healthy control group, T2DM group and CKD group (all P<0.05), but the serum level of SFRP5 in DN group was significantly lower than that in healthy control group, T2DM group and CKD group (all P<0.05). The differences between the four groups in the four aforementioned indicators were all statistically significant (all P<0.001). The Pearson correlation analysis showed that there was a significant correlation between the above four indicators (all P<0.01). The area under the ROC curve of TFF3, SFRP5, Gal-3, and NES-1 was 0.849, 0.807, 0.882, and 0.841 respectively. The area under the curve diagnosed by the combination of four indicators (0.986) was significantly higher than that of a single indicator, and the difference was statistically significant ( Z=3.75, 4.08, 3.63, 4.06, all P<0.05). Conclusions:The joint prediction model based on serum TFF3, SFRP5, Gal-3, and NES-1 can effectively improve the diagnostic accuracy of DN and provide an important basis for clinical diagnosis of DN.
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Objective To investigate the effects of trefoil factor 3 (TFF3) on the proliferation and apoptosis of human thyroid papillary carcinoma cell line (TPC-1) and its molecular mechanism. Methods The lentiviral expression vector of overexpression and knockdown of TFF3 gene were constructed, 293T cell was packaged to produce lentiviral particles, virus solution was collected and transfected into TPC-1 cells, enhanced cell TFF3-TPC-1 and enhanced control group ConTFF3-TPC-1; silencing cell shRNA-TFF3-TPC-1 and silencing control cells shCon -TPC-1. Western blotting, and Real-time PCR were used to detect the expression of TFF3 protein and mRNA of four groups. Growth curve and colony formation assay were used to detect the proliferation. Flow cytometry was used to analyze the apoptosis level of the four groups; Western blotting and immunocytochemistry were used to detect apoptosis-related protein and pathway protein phosphatidylinositol 3-kinase/ protein kinase B (PI3K/ Akt), nuclear factor-κB (NF-κB) expression. Results 1. Overexpression and inhibition of expression of TFF3 stable cell TFF3-TPC-1 and shRNA-TFF3-TPC-1 were constructed suscessfully. 2. The proliferation and cloning ability of TFF3-TPC-1 cells were significantly higher than those of ConTFF3-TPC-1 cells(P<0. 05 or P< 0. 01), the proliferation and cloning ability of shRNA-TFF3-TPC-1 cells were significantly lower than those of shCon-TPC-1 cells(P<0. 01); 3. The apoptosis rate of TFF3-TPC-1 cells was lower than that of ConTFF3-TPC-1 (0. 75%±0. 08% vs 5. 62%±0. 3%, P<0. 01),and the apoptosis rate of shRNA-TFF3-TPC-1 was higher than that of shConTPC-1 (22. 2% ± 1. 2% vs 5. 34% ± 0. 4%, P<0. 01); 4. After silencing TFF3 gene, the expressions of Bax, cytochrome C (Cyt-C), cleaved-Caspase-9, cleaved-Caspase-3 were up-regulated, and the expressions of Bcl-2, Akt, p-Akt and NF-κB-P65 were down-regulated (P<0. 05 or P<0. 01). Conclusion TFF3 may regulate the proliferation and apoptosis of TPC-1 cells by affecting the PI3K/ Akt/ NF-κB signaling pathway.
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Objective To evaluate the relationship between the Trefoil factor 3 (TFF3) serum,concentration and gastrointestinal failure(GIF) and discuss eaely diagnosis,treatment and prognosis in patients with GIF after esophageal cancer surgery.Methods To test the TFF3 levels of the serum during the postoperation of esophageal cancer by ELISA.Results Serum TFF3 concentrations measured prior to the occurrence of GIF were significantly higher than in control group (P ＜ 0.01).serum TFF3 concentration was significantly related to gastrointestinal tract function score(r =-0.712).Cox proportional hazards model analysis showed that the serum TFF3 concentrations at the time of occurrence of gastrointestinal failure,and 48 hours later,could be used as prognostic factors in critically ill pediatric patients with GIF(r =1.443 and 1.872,respectively).Conclusion TFF3 may play an important role in predicting GIF in pediatric critical illness and has a protective function in the mucosal repair process.
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Background:Gastric cancer is the second most frequently diagnosed cancer and the third cause of cancer death in China. Selection of candidates in high risk of gastric cancer by a simple and non-invasive marker with high sensitivity and then undergoing endoscopy is an optimal approach for large scale gastric cancer screening. Aims:To evaluate the clinical value of using trefoil factor 3(TFF3)as a serum biomarker for gastric cancer screening. Methods:Serum samples of 49 gastric cancer patients and 29 healthy subjects were collected for measurements of serum TFFs by ELISA from Jul. 2013 to Jan. 2014 at Ningbo Medical Treatment Center Lihuili Hospital. ROC curve and the area under curve(AUC)were used to verify the diagnostic performance of serum TFF1,TFF2 and TFF3 for gastric cancer. Correlations between TFFs and clinicopathological characteristics of gastric cancer were further analyzed. Results:Serum concentration of TFF3 in gastric cancer group was significantly higher than that in healthy controls[(43. 57 ± 19. 49)ng/ mL vs.(29. 97 ± 14. 20)ng/ mL, P 0. 05). AUC of serum TFF1,TFF2 and TFF3 for diagnosis of gastric cancer were 0. 56,0. 56 and 0. 83, respectively,which indicated that the performance of TFF3 was the best. Taken 33. 0 ng/ mL as the cut off value of TFF3, the sensitivity and specificity were 63. 3% and 82. 8% ,respectively,and the odds ratio for predicting gastric cancer was 8. 27. Significant correlations were existed between serum concentration of TFF3 and TNM stage,differentiation and lymph node metastasis of gastric cancer(P < 0. 05). Conclusions:Serum TFF3 is a promising non-invasive biomarker for gastric cancer screening.
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Background:TLR4 can mediate immune and inflammatory responses,TFF3,MUC2 are the intestinal mucosa protection factor and can maintain the intestinal barrier function. Aims:To investigate the effect of Anchang Yuyang decoction on colon tissue TFF3,MUC2 and TLR4 gene expressions in rats with ulcerative colitis. Methods:TNBS was used to establish ulcerative colitis model in rats. Ninety Wistar rats were randomly divided into normal control group,model group,low,moderate and high dose Anchang Yuyang decoction groups and mesalazine group,and distilled water,different concentrations of Anchang Yuyang decoction and mesalazine were given respectively. All the rats were sacrificed after 21 days. Colonic histopathological score was assessed,and RT-PCR was used to detect gene expressions of colon tissue TFF3, MUC2 and TLR4. Results:Compared with model group,histopathological score and TLR4 expression were significantly decreased in moderate,high dose Anchang Yuyang decoction groups and mesalazine group(P < 0. 05),expressions of TFF3 and MUC2 were significantly increased( P < 0. 05). Compared with moderate dose Anchang Yuyang decoction group,histopathological score in high dose Anchang Yuyang decoction group and mesalazine group was significantly decreased(P < 0. 05),and TFF3 expression was significantly increased( P < 0. 01). Compared with moderate dose Anchang Yuyang decoction group and mesalazine group,MUC2 expression in high dose Anchang Yuyang decoction group was significantly increased(P < 0. 01),and TLR4 expression was significantly decreased( P < 0. 01). Conclusions:Anchang Yuyang decoction can promote the repair of colonic mucosa in rats with ulcerative colitis,and its mechanism may be related to the increase of TFF3 and MUC2 gene expressions and down regulation of TLR4 gene expression.
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Objective To investigate the role of neuropeptide trefoil factor 3 (TFF3) in cocaine reward responses and the underlying mechanism.Methods Fifty SD rats were divided into 6 groups including Control group,Cocaine group,Cocaine + TFF3 (0.01 mg/kg) group,cocaine + TFF3 (0.1 mg/kg) group,cocaine + TFF3 (0.5 mg/kg) group and TFF3 (0.1 mg/kg) group,ip,n=7 ～ 9),and were treated with TFF3 / saline (ip),30 min later,rats were injected with cocaine (10 mg/kg ip) followed by 1 h hypedocomotion test.Immediately after behavioral test,rats (n=4～6) were decapitated and tissues of nucleus accumbens (NAc) were isolated and prepared for neurotransmitters analysis by HPLC; Another twenty one rats were divided into control and TFF3-treatment group,and the rats were trained with a modified 2-day cocaine CPP conditioning procedure.During cocaine CPP conditioning process,saline or TFF3 (0.1 mg/kg ip) was injected 30 min prior to cocaine injection (5 mg/kg ip).Results Systemic administration of TFF3 (0.1 mg/kg,ip) significantly increased the cocaine-induced locomotor activity (Total distance in 1 hour were (180±41) cm,(359±53) cm,(590±75) cm,(153±27) cm for Control,Vehicle + Cocaine,TFF3+Cocaine and TFF3+Vehicle groups respectively) and augmented cocaine rewarding effects in CPP(Post-training CPP score were (98± 18) s,(187±24) s for Vehicle + Cocaine,TFF3+Cocaine groups respectively).TFF3 (0.1 mg/kg ip) administration increased the dopamine concentration in the NAc induced by cocaine injection ((0.65±0.1) ng/ml,(1.24±0.14) ng/ml,(1.75±0.23) ng/ml,(0.74±0.21) ng/ml for Control,Cocaine,TFF3 + Cocaine and TFF3 + Vehicle groups respectively).Conclusion TFF3 is involved in regulation of behavioral response to cocaine,which is associated with the increasing of dopamine in the NAc.
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Objective To investigate the distribution and expression of trefoil factor 3(TFF3) in the eosinophilic cells and basophilic cells in the pars distalis of the rat pituitary .Methods The immunohistochemiscal staining technique was used to show the coexpression of TFF3/growth hormone (GH),TFF3/prolactin (PRL),TFF3/thyrotroph (TSH), TFF3/adrenocorticotropin (ACTH),TFF3/follicle stimulating hormone (FSH),TFF3/luteinizing hormone (LH) in two contiguous slices .Results The immunoreactive products of TFF 3 and chromophil cells were brown granules , mainly expressed in cytoplasm .ACTH positive signals were expressed in the cell membrane and mainly located in the pars distalis hypophyseos.TFF3 existed in parts of GH,PRL,TSH,ACTH,FSH and LH cells in contiguous slices, accounting for 19.4%, 22.4%, 9.2%, 6.5%, 35.7%, 8.3%, respectively , in which FSH was the most , PRL and GH were less . Conclusion TFF3 expresses in GH, PRL, TSH, ACTH, FSH, and LH cells in the pars distalis , which enriches the morphological data for the location of TFF 3 in gland cells of pars distalis hypophyseos .
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BACKGROUND:The intestine is not only the main target attacked by sepsis but also the vital organ which mediated sepsis. The recovery of the damaged intestinal barrier structure and function is related to the occurrence and outcome of multiple organ dysfunction syndrome (MODS). How to protect and reduce the damage of the intestinal mucosa and how to promote the reconstruction of the intestinal mucosa have been the important topics in sepsis for many years. This study aimed to investigate the influential factors of intestinal mucosal reconstruction after intestinal epithelial injuryin vivo in a mouse model of sepsis.METHODS:Mice were subjected to cecal ligation and puncture (CLP) for induction of sepsis to assess intestinal mucosal damage, epithelial cell apoptosis, and transformed number of goblet cells, and to detect the concentration of TNF-α, IL-1 and TGF-β1 and TFF3 (trefoil factor 3) expression in the small intestinal mucosa. All above were performed by HE staining, western blot, ELISA and immunohistochemistry respectively. The experimental animals were divided into a sepsis group and a sham-operation group. The animals with sepsis were separately killed at 6 (7 animals), 24 (7 animals) and 48 hours (7 animals) after CLP.RESULTS:Injured intestinal mucosa was observed in the 3 groups under a light microscope, in which damage scores in the 24-hour and 48-hour groups were higher than in the 6-hour group and no difference was found between the two groups. Moreover, less of goblet cells or other epithelial cells adjacent to the injured surface migrated into the wound to cover the denuded area. The number of goblet cells was substantially decreased in the three CLP groups compared with the sham-operation group. Protein levels of IL-1 and TNF-α were significantly increased by 3-4 fold at all time points when compared with the sham-operation group, and cleaved caspase-3 by 4 fold. Although TFF3 expression was modestly increased for 6 hours after the onset of CLP, it appeared to decline at 24 hours and 48 hours as shown by Western blot. A similar tendency was observed upon TGF-β1, i.e. the protein level was not elevated at 24 hours and 48 hours, but increased modestly at 6 hours.CONCLUSIONS:Sepsis from CLP shows less restitution on the surface of injured intestinal mucosa. There is evidence that both constant inflammatory reaction and epithelial cell apoptosis may affect mucosal reestablishment of the intestine at the onset of sepsis. Mucosa after severe sepsis showed the state of high inflammation, and declined goblet cell function and mucosal reconstruction, which affected the repair of damaged intestinal barrier. Constant inflammatory reaction, and declined goblet cell function and mucosal reconstruction ability may affect the reestablishment of intestinal mucosa at the onset of sepsis.
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Objective To investigate the expression and significance of trefoil factor 3 (TFF3) and stromal cell de-rived factor-1 (SDF-1) in papillary thyroid carcinoma (PTC) and adjacent tissues, and to provide the basis for clinical diagno-sis and prognosis evaluation. Methods The expressions of TFF3 and SDF-1 proteins in PTC and adjacent tissues were de-tected by immunohistochemical SP method. The relationship between expressions of TFF3, SDF-1 and clinicopathological features was analyzed. Results (1) The positive rate and the expression level of TFF3 protein were significantly higher in PTC than those in adjacent tissues, which were higher in patients with lymph node metastasis than those in patients without lymph nodes metastasis, and were higher in PTC patients of stageⅢ/Ⅳthan those in patients of stageⅠ/Ⅱ(P＜0.05). The strong positive rate of TFF3 protein in stageⅢ/Ⅳof PTC patients was higher than that in stageⅠ/Ⅱ(P＜0.01). (2) The posi-tive rate and the expression level of SDF-1 protein were significantly higher in PTC than those in adjacent tissues, which were higher in patients with lymph node metastasis than those in patients without lymph nodes metastasis, and were higher in PTC patients of stageⅢ/Ⅳthan those in patients of stageⅠ/Ⅱ, and were significantly higher in patients over 45 years old than those of patients≤45 years old (P＜0.05). (3) There was a positive correlation between expression levels of TFF3 and SDF-1 proteins in PTC (r=0.266,P＜0.05). Conclusion The higher expressions of TFF3 and SDF-1 in PTC were closely correlated with carcinogenesis and progression, which may play a significant value in judging the malignant degree and pro-gression of PTC.
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ObjectiveTo investigate the unfavorable factors of intestinal mucosa repair after the intestinal epithelial injury in vivo in a mouse model of sepsis. MethodsThe method of cecal ligature and puncture (CLP) was used to induce sepsis and then the intestinal mucosa damage, epithelial cell apoptosis and the number of transformed goblet cells were observed, and the concentrations of serum TNF-αt, IL-1 and TGF-β1 and TFF3 ( trefoil factor 3) in small intestinal mucosa were determined. All above various laboratory examinations were made by different assays including H-E staining, western blot, ELISA and immunohistochemistry respectively. The experimental mice were divided into sepsis group and sham operation control group. The mice with sepsis were separately sacrificed 6 hours ( n = 7 ), 24 hours ( n = 7) and 48 hours ( n = 7) after CLP. Results In septic mice group, the injured intestinal mucosa was found 6 hours after CLP. The damage scores in mice 24 h and 48 h after CLP were higher than those 6 h after CLP, but there was no significant difference between those 24 h and 48 h after CLP. Moreover, a few goblet cells or other epithelial cells adjacent to the injured surface migrated onto the wound to cover the denuded area. The number of goblet cells was substantially decreased in mice of sepsis group 6 hours after CLP compared with sham operation control group. Compared with sham operation control group, levels of IL-1 and TNF-α significantly increased 3-4 times in mice of sepsis group at all intervals, and the phosphorylated caspase-3 increased 4 times. Although TFF3 assayed by using Western blot showed modest increase 6 h after CLP and it declined 24 h and 48 h later. A similar change was found in TGF-β1, it modestly increased 6h after CLP, but it didn't elevate 24 h and 48 h later. ConclusionsSevere sepsis keeps on the inflammatory reaction and epithelial cell apoptosis, preventing the repair of intestinal mucosa from injury.
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Objective To investigate immunological dysfunction of intestine mucosa barrier in a rat model of sepsis. Method Sixty Sprague-Dawley rats were assigned randomly(random number) into sepsis group (n = 45)and control group (n = 15). The animals in sepsis group were subjected to cecal ligation and puncture (CLP), whereas rats of control group underwent a sham surgery. The ileac mucosa and segments were harvested 3 h, 6 h and 12 hours after CLP, and the blood samples were collected. Pathological changes, protein levels of defensin-5 (RD-5) and trefoil factor-3(TFF_3) mRNA, lymphocytes apoptosis in the intestinal mucosa were determined. In an additional experiment, the gut-origin bacterial DNA in blood was detected. Results In the septic animals, in-testinal mucosa showed marked injury with loss of ileal villi, desquamation of epithelium, detachment of the lamina propria, hemorrhage and ulceration. Compared with control, the expression of TFF_3 mRNA and level of RD-5 pro-tein were decreased and the mucosal lymphocyte apoptosis increased (P < 0.05) in sepsis group. Compared with control group, the significant differences in RD-5 and TFF_3 mRNA appeared 3 hours after CLP and those differ-ences were progressively increased in 6 hours and 12 hours after CLP in sepsis group (P < 0.05, F of RD-5 = 11. 76, F of TFF_3 = 16.86 and F of apoptosis = 122.52). In addition, the gut-origin bacterial DNA in plasma de-tected was positive in all sepsis animals. Conclusions It suggests that immunological function of intestinal mucosa is impaired in septic rats and further worsened following the course of sepsis.
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Objective: To investigate the changes of TFF2 and TFF3 expression in sublingual gland during spontaneous healing of experimental gastric ulcer in rats. Methods: A total of 48 male SD rats were divided into gastric ulcer group (n = 42, ulcers were induced by injection of acetic acid to the submucosal of paries anterior gastricus) and normal group (71 = 6). Immunohistochemical and RT-PCR methods were used to examine the expression of TFF2 and TFF3 in the sublingual gland in gastric ulcer group and normal group. Results: Immunohistochemical staining showed that positive signals of TFF2 and TFF3 were mainly located in the striated duct, intercalated duct, myoepithelial cells, and some lumens. Compared with the control group, the integrated optical density (IOD) value of TFF2 was obviously increased on day 1 after gastric ulcer(P<0.01), and reached the bottom on day 2, then gradually increased again on day 4, 6 (P<0.01), and kept at a high level during day 10-23 (P<0.05). The IOD value of TFF3 was similar to that of the normal group on day 1, 2 after gastric ulcer, then gradually increased on day 4, 6 (P<0.05), reached the peak on day 10 after gastric ulcer (P<0.01), and kept at a high level till day 23 after gastric ulcer(P<0.05). The results of RT-PCR showed that the changes of TFF2 mRNA and TFF3 mRNA were basically consistent with the change of their corresponding peptides. Conclusion: TFF2 and TFF3 expression is increased in the sublingual gland during the spontaneous healing of experimental gastric ulcer in rats, and they may participate in the healing process.