RESUMO
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.
RESUMO
Objective:To investigate the relationship between caspase recruitment domain protein 9 (CARD9) gene polymorphism and the risk of sepsis related liver injury.Methods:A case-control study was conducted to recruit 122 patients with sepsis in intensive care unit (ICU). The septic patients with liver injury were divided into the liver injury group ( n=66), and the patients without liver injury were set as the control group ( n=60). TaqMan genotyping technique was performed to detected the six single nucleotide polymorphisms of CARD9 gene in peripheral blood, and the baseline data of the two groups were observed. The differences of genotype distribution and allele frequency between the two groups were compared, and the odds ratios ( OR) were calculated. Kolmogorov-Smirnov test was used for normal distribution test, the measurement data that do not conform to the normal distribution are expressed in M ( Q1, Q3), and the Mann Whitney U test was used for the comparison between the two groups. The comparison of counting data between groups was adopted χ 2 inspection and Fisher exact probability method. χ 2 test is used to determine whether the genotype distribution conforms to hardy Weinberg equilibrium test. Results:Compared with the control group, the CC genotype distribution of rs10781500 locus of CARD9 gene in liver injury group was significantly increased (19 vs. 36, χ 2=6.87, P=0.033), and sepsis patients with C allele are more likely to induce liver injury ( OR=1.375, 95% CI 1.024-1.846, P=0.023). Patients with CC genotype have a higher risk of liver injury ( OR=2.696, 95% CI 1.238-5.869, P=0.012). In the liver injury group, the alanine aminotransferase and SOFA score in CC genotype at re10781500 locus were significantly higher than those in non-CC genotype (310 (213, 648) U/L vs. 187 (114, 304) U/L, Z=3.32, P=0.001 and 9 (6, 14) points vs. 7 (5, 9) points, Z=2.49, P=0.013). Conclusion:The CC genotype at rs10781500 locus of CARD9 gene was associated with sepsis-induced liver injury, and the C allele may be a susceptibility gene.
RESUMO
Objective:To investigate the role of caspase recruitment domain protein 9(card9) from macrophage in pancreatic acinar-to-ductal metaplasia.Methods:Card9 siRNA1, card9 siRNA2 and card9 siRNA3 were constructed; fluorescence microscopy was used to investigate the fluorescence intensity of macrophages, and real-time quantitative PCR method was performed to detect the expressed level of card9 mRNA to obtain the best transfection rate. 100 μg/ml β glucan was added into 5×10 5 macrophages in vitro culture for 12 or 24 hours, which were divided into positive group (macrophages), β glucan-stimulated positive group (β dextran+ macrophage), negative group (card9 -/- macrophage) and β glucan-stimulated negative group (β dextran+ card9 -/- macrophages). Western blotting was applied to determine the protein level of card9 in macrophages. Then, 1×10 5 macrophages and 1×10 5 pancreatic acinar cells were co-cultured in upper and lower transwell chamber in vitro for 120 hours, which were divided into positive group (macrophages+ acinar cells), 100 μg/ml and 500 μg/ml β glucan-stimulated positive group, negative group (card9 -/- macrophage+ acinar cell), 100 μg/ml and 500 μg/ml β glucan-stimulated negative group. Pancreatic acinar cells in the lower chamber were collected and immunofluorescence was applied to assay the duct metaplasia marker CK19 protein expression. Results:At 24 hours of transfection using siRNA, the intracellular fluorescence intensity in macrophages reached a peak. Card9 siRNA at the concentration of 200 nmol/l showed the highest interference efficiency. Card9 protein in positive group, β glucan-stimulated positive group, negative group, and β glucan-stimulated negative group were 0.81±0.05, 1.46±0.05, 0.42±0.06 and 0.46±0.06, respectively; card9 expression in β glucan-stimulated positive group was obviously higher than that in positive cell group, and the difference was statistically significant ( P<0.05). Finally, after 100 or 500 μg/ml β glucan stimulation, the green fluorescence in pancreatic acinar cells increased significantly compared with positive group, exhibiting β glucan concentration dependence. Conversely, CK19 protein in negative group and 100 and 500 μg/ml β glucan-stimulated negative group was obviously decreased compared with positive group. Conclusions:The expression level of card9 in macrophages can induce acinar-to-ductal metaplasia, indicating that card9 may mediate in the pathogenesis of pancreatic cancer.
RESUMO
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.