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Objective:To investigate the effects of telomerase reverse transcriptase (TERT) on neutrophils apoptosis in rats with acute necrotizing pancreatitis (ANP).Methods:Twenty-four Sprague Dawley (SD) rats were randomly divided into three groups including control group, ANP (3h、6h) group and TERT inhibitor(BIBR1532)group using random number method with 6 in each group. ANP rats were induced by retrograde injection of 5% sodium taurocholate into the pancreaticcobiliary duct. After 3 and 6 hours, the fresh neutrophils were collected and isolated from peripheral blood of ANP rats. Rats were intraperitoneally injected with 2 mg/kg BIBR1532. After ANP modeling for 3 h, rats were killed and peripheral neutrophils were collected. Subsequently, the expression of TERT mRNA in neutrophils was tested by real-time quantitative PCR; the protein levels of TERT, BCL-xL and Bax were determined by Western blotting; neutrophil apoptosis was detected by flow cytometry; TNF-α and IL-6 were assayed by Elisa; the rat pancreatic tissue was pathologically examined.Results:In neutrophils from control group, ANP 3 h group, ANP 6 h group and BIBR1532 group, TERT mRNA was 1.03±0.26, 3.31±1.07, 5.21±0.78 and 1.95±0.49; TERT protein expression was 0.09±0.03, 0.43±0.12, 0.58±0.11 and 0.22±0.07; Bcl-xL protein expression was 0.19±0.05, 0.50±0.07, 0.85±0.04 and 0.40±0.11; Bax protein expression was 0.29±0.08, 0.23±0.03, 0.17±0.02 and 0.43±0.12; apoptosis rate was 10.03±0.74%, 7.99±0.27%, 6.65±0.36% and 22.98±2.86%. TERT mRNA and protein expression and Bcl-xL protein expression in ANP 3 h and 6 h group were higher than those in control group, but Bax preotein expression and apoptosis rate were lower than those in control group; TERT mRNA and protein expression and Bcl-xL protein expression in BIBR1532 group were lower than those in ANP 3 h group, but Bax protein expression and apoptosis rate in BIBR1532 group were higher than those in ANP 3 h group; and all the differences were statistically significant (all P value <0.05). The level of TNF-α was [(96.67±27.12)ng/L, (382.30±46.33)ng/L and (206.88±36.42)ng/L], IL-6 was [(43.34±14.50)ng/L, (134.21±16.13)ng/L and (88.06±13.05)ng/L in control, ANP 3 h and BIBR1532 group; those in ANP 3 h group were all higher than those in control group, but those in BIBR1532 group were lower than those in ANP 3 h group; all the differences were statistically significant (all P value <0.05). Compared with ANP group, the morphology of pancreatic tissue was obviously alleviated in BIBER1532 group. Conclusions:TERT expression is obviously increased in peripheral neutrophils in ANP rats, and apoptosis rate of neutrophils from ANP rats is greatly increased after TERT inhibitor treatment, demonstrating that TERT could inhibit the apoptosis of peripheral neutrophil in ANP rats.
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Objective:To investigate the relationship between caspase recruitment domain protein 9 (CARD9) gene polymorphism and acute pancreatitis (AP) and the clinical efficacy of somatostatin.Methods:A total of 86 patients with AP treated in Shanghai Songjiang District Central Hospital from June 2019 to may 2020 were selected as the research object, and 81 healthy volunteers were selected as the control group for a prospective cohort study. The nucleotide database of National Center for Biotechnology Information (NCBI) was consulted to screen 10 common single nucleotide polymorphisms of CARD9.The single nucleotide polymorphism of CARD9 was detected by SNapShot micro sequencing. All patients with AP were treated with somatostatin. The relationship between CARD9 single nucleotide polymorphism and clinical symptoms and auxiliary examination indexes was observed.The measurement data of normal distribution were compared by independent sample t-test. The measurement data of non normal distribution are represented by M (Q1, Q3), and the rank sum test is used for comparison between groups. The comparison of counting data between groups was adopted χ 2 inspection. Results:The frequency of CARD9 rs10870077 C>G SNP in patients of AP group was significantly higher than that in healthy controls ( OR=1.934, 95% CI=1.011-3.700, P=0.046). Compared with CC genotype, the disappearance time of abdominal pain and abdominal distension in the somatostatin treatment group of CARD9 rs10870077 C>G moderate and severe AP patients was significantly longer ((5.64±2.06) d and (3.76±1.23) d, t=2.98, P=0.006), and the average hospital stay in the somatostatin treatment group of CARD9 rs10870077 C>G severe AP patients was increased by ((13.25±5.31) d and (9.00±3.68) d, t=1.51, P=0.170). Conclusion:CARD9 rs10870077 C>G is a predisposing factor for AP, which is related to the individual differences in the clinical efficacy of somatostatin in severe AP.
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Objective:To evaluate the correlation between Card 9 gene polymorphism and acute pancreatitis(AP).Methods:70 AP patients and 70 healthy subjects from Shanghai Songjiang District Central Hospital from June 2019 to February 2020 were enrolled. TaqMan probe method was used to assay genotype distributions of the Card 9 polymorphisms rs10870077, rs4077515, rs10781499, rs141992399, rs139265120. Real-time quantitative PCR was used to determine the level of Card 9 mRNA, electrochemiluminescence immunoassay was applied to assay IL-6 and procalcitonin, and nephelometry was performed to measure the C-reactive protein(CRP).Results:Compared with the control group, the genotype and allele frequency of Card 9 gene rs10870077 C>G were significantly elevated in AP patients with statistically significant difference (31.4% vs 50.0%, P<0.05). There was no statistically significant difference on the allele frequency of Card 9 rs4077515AG and rs10781499AG, especially on rs141992399 C>G and rs139265120 A>G. C>Gpolymorphism in Card 9 rs10870077 resulted in an obvious increase of serum Card 9 mRNA expression in AP patients from 3.90±1.96 to 6.20±2.82, and the difference was statistically significant ( P<0.05), but there was no statistically significant difference on the Card 9 mRNA between AP patients with Card 9 rs4077515AG and rs10781499AG and the controls. IL level in AP patients with Card 9 rs10870077CG was greatly higher than that in those with Card 9 rs10870077 CC and GG [(614.7±1531.8 ng/L vs (372.5±1127.9)and (385.5±598.7)ng/L]. But compared with GG genotype, CRP level was obviously decreased [(34.84±50.64)mg/L vs (55.30±87.02)mg/L], and the procalcitonin was obviously increased [(1.98±4.70)μg/L vs (0.77±1.12)μg/L], and all the differences were statistically significant (all P value<0.05). Conclusions:Card 9 rs10870077 C>G mutation could upregulate the expression of Card 9 mRNA and increase IL-6 level, which may be a high risk for the occurrence of AP patients.
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Objective To investigate dynamic changes of serum Tau proteins and their correlation with cognitive dysfunction in patients with acute traumatic brain injury (TBI).Methods A total of 95 patients with acute TBI were retrospectively studied by case-control study.There were 61 males and 34 females,with age of 16-65 years [(40.7 ± 13.6)years].The Glasgow coma scale (GCS) was 3-8 points in 9 patients,9-12 points in 11,and 13-15 points in 75.A total of 30 healthy physical examinees were recruited as control group.The levels of Tau proteins were measured at days 1,3,5,7 and 14 after TBI.The cognitive dysfunction was evaluated by the Montreal Cognitive Assessment (MoCA) score at 6 months after injury.The correlation between Tau protein levels at different time points and MoCA was determined.Results The serum Tau proteins of TBI group was significantly higher than that of control group at all time points (P ＜ 0.05).In TBI group,39 (41％) out of 95 patients developed cognitive dysfunction assessed by MoCA scale.The main manifestations of cognitive dysfunction were the defects in visual spatial and acting function,delayed memory,language,abstract,attention and calculation,with statistical significance compared with control group (allP ＜ 0.05).The serum Tau proteins of patients with cognitive dysfunction were significantly higher than those without cognitive dysfunction at all time points after TBI (P ＜ 0.05).Tau proteins at days 1,3,5 after TBI was significantly correlated with cognitive dysfunction at 6 months after TBI (P ＜ 0.05).Conclusions The levels of serum Tau proteins show a significant increase after TBI,the early changes of which are statistically related to cognitive dysfunction.The early changes of serum Tau protein after TBI can be used as a reliable biomarker for early prediction of cognitive function prognosis.
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Objective To investigate the EUS elastographic patterns of normal pancreas and focal pancreatic lesions, and its value for characterizing and differentiating between benign and malignant pancreatic tissues. Methods Between January and June 2009, 6 patients with normal pancreas and 9 patients with focal pancreatic lesions were enrolled. Real-time elastography was carried out during the conventional EUS examination. The elastographic images were scored with 1 to 5 based on the elastographic pattern. Results Nine focal pancreatic lesions were finally diagnosed as pancreatic cancer (n = 4), cyst-adenocarcinoma (n =1), cyst-adenoma (n = 2) and focal pancreatitis (n = 2), respectively. Pancreatic cancers were scored as 3(n = 1) and 4 (n = 3). Cyst-adenocarcinoma was scored as 5 and cyst-adenomas were scored both as 2,while focal pancreatitis were scored as 2 and 3, respectively. Elastographic pattern was scored as 1 (n = 5)and 2 (n = 1) in 6 normal pancreas. When scores 1 and 2 were assigned to benign lesion and 3 to 5 to malignancy, the overall diagnostic accuracy of EUS elastography for focal pancreatic lesions was 88. 9%(8/9). Conclusion There are apparent differences in elastographic patterns between benign and malignant pancreatic tissues. EUS elastography is a promising method that allows characterization and differentiation of benignancy and malignancy.
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AIM:To study the effect of intestinal endotoxemia(IETM) on hepatic energy metabolism in acute liver failure. METHODS:Intoxication by thioacetamide (TAA) was used to establish rat model of acute liver injury.Ketone body(acetoacetate and ?-hydroxybutyrate) in arterial blood and ATP content of hepatocellular mitochondria were determined by using enzymatic fluorimetric micromethod.Colectomy was adopted in observing the changes in plasma endotoxin content and serum alanine aminotransferase (ALT) activity. RESULTS:In the TAA group,plasma endotoxin content and serum ALT activity were all significantly higher than those in the control group(P