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Objective:To investigate the value of serum soluble programmed cell death protein 1 (sPD-1), soluble B7 homolog 5 (sB7-H5) and trefoil factor 2 (TFF2) in evaluating the severity of disease and the risk of death in patients with acute pancreatitis (AP).Methods:A prospective research method was adopted. Three hundred and twenty-eight patients with AP (AP group) from February 2020 to February 2021 in Xiangyang Central Hospital were selected, including 124 patients with mild AP (MAP), 106 patients with moderately severe AP (MSAP) and 98 patients with severe AP (SAP). The serum levels of sPD-1, sB7-H5 and TFF2 were measured by enzyme-linked immunosorbent assay and compared with 60 healthy people (healthy control group). The patients with AP were followed up for 90 d, 284 patients survived and 44 died. The amylase, C-reactive protein (CRP), procalcitonin (PCT), acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ), sequential organ failure assessment (SOFA), modified CT severity index (MCTSI), sPD-1, sB7-H5 and TFF2 were compared between the two groups. Pearson method was used for correlation analysis. Multivariate Logistic regression was used to analyze the independent risk factors of death in patients with AP. The efficacy of sPD-1, sB7-H5 and TFF2 in predicting the death in patients with AP was evaluated using the receiver operating characteristics (ROC) curve.Results:The sPD-1, sB7-H5 and TFF2 in AP group were significantly higher than those in healthy control group: (177.99 ± 17.81) ng/L vs. (50.20 ± 10.81) ng/L, (2.69 ± 0.72) μg/L vs. (1.40 ± 0.35) μg/L and (569.97 ± 38.91) μg/L vs. (94.59 ± 11.98) μg/L, and there were statistical differences ( P<0.01). The amylase, sPD-1, sB7-H5 and TFF2 in patients with MSAP and SAP were significantly higher than those in patients with MAP: (639.36 ± 91.67) and (835.24 ± 109.30) U/L vs. (575.24 ± 89.78) U/L, (180.13 ± 20.61) and (221.17 ± 15.70) ng/L vs. (142.03 ± 16.76) ng/L, (2.85 ± 0.74) and (3.34 ± 0.82) μg/L vs. (2.05 ± 0.52) μg/L, (539.66 ± 36.58) and (763.55 ± 40.08) μg/L vs. (442.90 ± 35.79) μg/L, the indexes in patients with SAP were significantly higher than those in patients with MSAP, and there were statistical differences ( P<0.01). Pearson correlation analysis result showed that sPD-1 was positively correlated with sB7-H5 and TFF2 in patients with AP ( r = 0.552 and 0.641, P<0.01), and the sB7-H5 was positively correlated with TFF2 ( r = 0.610, P<0.01). The amylase, CRP, PCT, APACHE Ⅱ, SOFA, MCTSI, sPD-1, sB7-H5 and TFF2 in the dead patients were significantly higher than those in the living patients: (1 098 ± 105) U/L vs. (641 ± 93) U/L, (235.60 ± 40.17) mg/L vs. (118.04 ± 32.90) mg/L, (4.32 ± 0.52) μg/L vs. (3.14 ± 0.44) μg/L, (19.39 ± 3.14) scores vs. (11.18 ± 2.53) scores, (12.13 ± 2.78) scores vs. (7.40 ± 2.15) scores, (7.12 ± 1.73) scores vs. (4.31 ± 1.52) scores, (222.23 ± 22.30) ng/L vs. (171.14 ± 18.50) ng/L, (3.37 ± 0.89) μg/L vs. (2.59 ± 0.59) μg/L and (629.27 ± 39.63) μg/L vs. (560.78 ± 30.45) μg/L, and there were statistical differences ( P<0.01). Multivariate Logistic regression analysis result showed that CRP, PCT, APACHE Ⅱ, SOFA, sPD-1, sB7-H5 and TFF2 were independent risk factors death of in patients with AP ( OR = 1.339, 1.416, 1.285, 1.327, 1.092, 1.171 and 1.080; 95% CI 1.145 to 1.566, 1.146 to 1.751, 1.132 to 1.460, 1.150 to 1.531, 1.024 to 1.164, 1.072 to 1.280 and 1.031 to 1.131; P<0.01). The ROC curve analysis result showed that the area under the curve of sPD-1, sB7-H5 and TFF2 combined detection to predict the death in patients with AP was larger than that of sPD-1, sB7-H5, and TFF2 alone detection (0.870 vs. 0.771, 0.734 and 0.685). Conclusions:The increase of serum sPD-1, sB7-H5 and TFF2 levels in patients with AP is related to the severity of disease of patients with AP. The combined detection of the indexes can assist in evaluating the risk of death in patients with AP.
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Background:Gastric cancer is one of the malignant tumors with highest morbidity and mortality,and the early change of molecular marker in gastric mucosal lesion is the hot spot of gastric cancer study. Aims:To investigate the expressions and clinical significance of trefoil factor 2(TFF2),claudin 18(CLDN18),mucin 5AC(MUC5AC)in gastric mucosal lesions. Methods:Gastroscopy biopsies and surgery specimens from Dec. 2008 to May 2009 at Inner Mongolia People’s Hospital were collected,including 20 normal gastric mucosal tissues,20 intestinal metaplasia tissues,11 dysplasia tissues and 20 gastric cancer tissues. The protein expressions of TFF2, CLDN18, MUC5AC were determined by immunohistochemical SP method. Results:The positive expression rates of TFF2,CLDN18 and MUC5AC in normal gastric mucosal tissues were all 100% ,and were gradually decreased in the order of intestinal metaplasia,dysplasia and gastric cancer tissue,the differences were statistically significant(P < 0. 01). Conclusions:Expressions of TFF2,CLDN18 and MUC5AC protein are closely related to the degree of malignancy of gastric mucosal lesions,and can be considered as a potential biological marker for predicting the development and prognosis of gastric cancer.
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Objective To over-express human trefoil factor 2 (hTFF2) by Escherichia coli system and an-alyze its activities in promoting migration and anchorage-independent growth in SW480 colonic cancer cells. Meth-ods hTFF2 gene encoding mature peptide was obtained by RT-PCR, and the recombinant expression vector pET32a-hTFF2 was constructed. Then pET32a-hTFF2 was transformed into E. coli BL21-32a and TrxA-hTFF2 fu-sion protein was induced to over-express. The expressed product was isolated by Ni-NTA affinity chromatography, purified by dialysis and identified by Western blotting. The activities of the recombinant hTFF2 in promoting SW480 cells migration and anchorage-independent growth were analyzed by MicroChemotaxis Chamber migration assay and Soft-agar assay,respectively. Results The TrxA-hTFF2 fusion protein was expressed to 220 mg/L at high purity. In vitro model demonstrated that recombinant hTFF2 obviously enhanced SW480 cell migration activity and anchor-age-independent growth. Conclusion The recombinant hTFF2 can be expressed in E. coli with high production, purity and biological activities. And its roles in cell migration and anchorage-independent growth suggest that up-regulation of TFF2 in colonic cancer might be involved in cancer invasion and metastases.
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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.