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Objective:To explore the value of serum activin A (ACT-A) level in early identification of moderate and severe acute pancreatitis (AP).Methods:A prospective case control study was conducted. A total of 120 patients with AP admitted to department of hepatobiliary surgery of Affiliated Nanhua Hospital of Hengyang Medical College of University of South China between October 2020 and April 2022 were recruited. According to the revised Atlanta classification, all patients were classified into mild AP group and moderate-to-severe AP group. The blood samples within 24 hours of onset were drawn, and the serum ACT-A and C-reactive protein (CRP) levels were detected by enzyme-linked immunosorbent assay (ELISA). The Ranson score and the modified CT severity index (MCTSI) were performed. Pearson correlation method was used to analyze the correlation of various parameters. The receiver operator characteristic curve (ROC curve) was plotted to analyze the predictive value of ACT-A and CRP for moderate-to-severe AP.Results:A total of 120 patients with AP were enrolled, including 83 patients with mild AP and 37 patients with moderate-to-severe AP. Serum ACT-A and CRP levels within 24 hours of onset in the moderate-to-severe AP group were significantly higher than those in the mild AP group [ACT-A (ng/L): 140.4±37.7 vs. 53.9±30.5, lg CRP: 1.42±0.91 vs. 0.77±0.70, both P < 0.01], and the Ranson score and MCTSI score were also significantly higher than those in the mild AP group (Ranson score: 5.3±1.3 vs. 1.8±1.6, MCTSI score: 5.5±1.0 vs. 2.7±1.2, both P < 0.01). Correlation analysis showed that the serum ACT-A level was positively correlated with serum CRP level, Ranson score and MCTSI score ( R2 value was 0.272, 0.841, 0.616, respectively, all P < 0.05). ROC curve analysis showed that the serum ACT-A, CRP and Ranson score had predictive value for moderate-to-severe AP. The area under the ROC curve (AUC) was 0.948 [95% confidence interval (95% CI) was 0.909-0.986], 0.711 (95% CI was 0.606-0.815), 0.946 (95% CI was 0.910-0.982), respectively. When serum ACT-A > 112.6 ng/L, the sensitivity and specificity of predicting moderate-to-severe AP were 78.38% and 96.39%, respectively, which was better than serum CRP with sensitivity and specificity of 72.92% and 66.27%, respectively, and the specificity was better than Ranson score (71.08%). Conclusion:ACT-A can be detected in the early stage of AP, and it is positively correlated with the disease severity, which can early identify moderate-to-severe AP.
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ObjectiveTo obtain HSC-T6 cells with stable expression of Cas9 protein and HSC-T6-COX-2-/- cells with COX-2 gene defect by transfecting HSC-T6 cells with CRISPR/Cas9 lentiviral vector, and to provide a good method for further functional research and new strategies for the clinical treatment of liver fibrosis. MethodsThe COX-2 gene-specific sgRNAs (COX-2-sgRNA-1, COX-2-sgRNA-2, COX-2-sgRNA-3) were designed, synthesized, and connected to the GV371 vector, and the recombinant plasmid and the packaging plasmid were transfected into 293T cells to form lentivirus particles; the fluorescence method was used to measure virus titer. The most appropriate amount of the virus was calculated based on MOI. Lenti-Cas9-puro was transfected into HSC-T6 cells, and HSC-T6-Cas9 cells were screened out by puromycin; Lenti-COX-2-sgRNA-EGFP was transfected into HSC-T6-Cas9 cells to obtain HSC-T6-COX-2-/- cells. Cruiser enzyme digestion and Western blot were used to verify gene knockout at the gene and protein levels. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsSequencing verified that the COX-2-sgRNA expression vector was constructed successfully. Recombinant expression plasmids and packaging plasmids were transfected into 293T cells to form lentivirus particles, and the fluorescence method showed a virus titer of ＞1×108. HSC-T6 cells with stable expression of Cas9 protein and HSC-T6-COX-2-/- cells with COX-2 gene defect were successfully constructed. The HSC-T6-Cas9 group had significantly higher relative mRNA expression of LV-Cas9-Puro than the CON group (541.93±105.76 vs 1.00±0.02, t=12.995, P＜0.01). Cruiser enzyme digestion and Western blot showed that the CRISPR/Cas9 lentivirus expression vectors played a role in the target, among which COX-2-sgRNA-2 knockout had the most significant effect, and this group had a significant reduction in the protein expression level of COX-2 compared with the CON group and the NC group (both P＜0.05), suggesting that COX-2-sgRNA was active. ConclusionA CRISPR/Cas9 lentivirus vector is successfully constructed for COX-2 target gene, and HSC-T6-COX-2-/- cells with stable COX-2 gene knockout are obtained.
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ObjectiveTo investigate the effect and significance of cyclooxygenase-2 (COX-2) inhibitors on the expression of the Acsl gene family in the ileum of rats with nonalcoholic fatty liver disease (NAFLD). MethodsA total of 45 Sprague-Dawley rats were randomly divided into normal control group (15 rats given normal diet), NAFLD model group (15 rats given high-fat diet), and nimesulide group (15 rats given high-fat diet and nimesulide). All rats were sacrificed after 12 weeks of feeding, and then blood samples were collected from the inferior vena cava to measure total cholesterol (TC) and triglyceride (TG). HE staining and oil red O staining were performed for the liver to evaluate the degree of hepatic steatosis in each group, and quantitative real-time PCR was used to measure the mRNA expression of the Acsl family genes in the ileum. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the normal control group, the NAFLD model group had significant increases in serum TC and TG and marked hepatic steatosis (all P＜0.05); compared with the NAFLD model group, the nimesulide group had significant reductions in serum TC and TG and degree of hepatic steatosis (all P＜0.05). Compared with the normal control group, the NAFLD model group had a significant increase in the expression of COX-2 in the ileum (P＜0.05), and compared with the NAFLD model group, the nimesulide group had a significant reduction in the expression of COX-2 in the ileum (P＜005). Compared with the normal control group, the NAFLD model group had significant increases in the mRNA expression of Acsl3 and Acsl5 in the ileum (both P＜0.05), and compared with the NAFLD model group, the nimesulide group had significant reductions in the mRNA expression of Acsl3 and Acsl5 (both P＜0.05). ConclusionThe COX-2 inhibitor nimesulide can regulate the expression of the Acsl gene family in the ileum of rats with NAFLD, suggesting that COX-2 inhibitors may inhibit the progression of NAFLD through the Acsl gene.
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Objective To study the radiation-induced mitochondrial damage and energy metabolic alteration in pancreatic exocrine cells,and to explore underlying mechanism.Methods Rat pancreatic exocrine cells (AR42 J) were divided into control group and experimental group irradiated with 6 Gy of X-rays.Mitochondrial membrane potential was detected at 24,48,72 and 96 h,the lactic acid and ATP production were detected at 24 h and 48 h,and reactive oxygen species (ROS) were detected at 24 h after irradiation.The expressions of energy metabolism related factors of HIF-1α,LDHA and PDH were detected by Western blot.The animal experiments were conducted to confirm the changes.According to random number table,eight rats were randomly divided into two groups with 4 rats in each.The irradiated group was exposed to 12 Gy of X-rays,while the control group sham-irradiated.Results Compared with the nonirradiated control group,the mitochondrial membrane potential (△Ψm) of the experimental group was progressively decreased at 24-96 h after irradiation (t =5.438-17.687,P＜0.05).The ATP content in the experimental group decreased at 24 h and 48 h (q=17.300,8.328,P＜0.05),the lactic acid increased (q =21.790,16.250,P＜0.05),and the ROS level increased (t =7.935,P＜0.05).The expressions of HIF-1α and LDHA were significantly increased,but PDH was reduced after radiation.Silencing HIF-1α by siRNA eliminated radiation-induced energy metabolic alteration.These changes were confirmed with animal experiments by locally irradiating rats.Conclusions The expression of HIF-1α is upregulated by irradiation,which leads to the change of energy metabolism as the enhancement of glycolysis pathway and the inhibition of aerobic oxidation of mitochondria in pancreatic exocrine cells.
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OBJECTIVE: To investigate the morphological and hemodynamic parameters associated with middle cerebral artery (MCA)bifurcation aneurysm rupture. METHODS: A retrospective study of 67 consecutive patients was carried out based on 3D digital subtraction angiography data. Morphological and hemodynamic parameters including aneurysm size parameters (dome width, height, and perpendicular height), longest dimension from the aneurysm neck to the dome tip, neck width, aneurysm area, aspect ratio, Longest dimension from the aneurysm neck to the dome tip (Dmax) to dome width, and height-width, Bottleneck factor, as well as wall shear stress (WSS), low WSS area (LSA), percentage of LSA (LSA%) and energy loss (EL) were estimated. Parameters between ruptured and un-ruptured groups were analyzed. Receiver operating characteristics were generated to check prediction performance of all significant variables. RESULTS: Sixty-seven patients with MCA bifurcation aneurysm were included (31 unruptured, 36 ruptured). Dmax (p=0.008) was greater in ruptured group than that in un-ruptured group. D/W (p<0.001) and the percentage of the low WSS area (0.09±0.13 vs. 0.01±0.03, p<0.001) were also greater in the ruptured group. Moreover, the EL in ruptured group was higher than that in un-ruptured group (6.39±5.04 vs. 1.53±0.86, p<0.001). Multivariate regression analysis suggested D/W and EL were significant predictors of rupture of MCA bifurcation aneurysms. Correlation analyses revealed the D/W value was positively associated with the EL (R=0.442, p<0.01). CONCLUSION: D/W and EL might be the most two favorable factors to predict rupture risk of MCA bifurcation aneurysms.
الموضوعات
Humans , Aneurysm , Angiography , Angiography, Digital Subtraction , Biomarkers , Hemodynamics , Intracranial Aneurysm , Middle Cerebral Artery , Neck , Retrospective Studies , Risk Assessment , ROC Curve , Ruptureالملخص
Objective To investigate the therapeutic effect of the ultra-early stent-assisted coil embolization of the ruptured intracranial aneurysms. Methods The clinical data of 13 patients with ruptured intracranial aneurysm treated by ultra-early stent-assisted coil embolization were analyzed retrospectively. The preoperative Hunt-Hess gradeⅠ-Ⅱ was in 7 cases,gradeⅢ was in 4 cases,and grade Ⅳ was in 2 cases. The patients were treated with stent-assisted coil embolization under the general anesthesia with endotracheal intubation within 24 h of aneurysm rupture. The postoperative embolization was assessed according to the Raymond grading standard. The postoperative complications and the assessment of the follow-up results from 1 to 6 months after procedure according to the modified Rankin scale (mRS ) scores were observed. Results All 11 patients recovered well,1 case had postoperative hemiplegia,1 case had postoperative bleeding,and none of them died. During the follow-up period,no patients had rebleeding, 1 had recurrence,and DSA revealed that the patient was embolized completely at 2 months after reembolization. Conclusion The method of ultra-early stent-assisted coil embolization of ruptured intracranial aneurysms is feasible. It may improve the cure rate of the ruptured aneurysms and improve the prognosis of patients.
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ObjectiveTo investigate the experience in the treatment of acute encephalocele associated with delayed intracranial hematoma during severe brain injury operation.Methods42 patients suffered from acute encephalocele during brain injury operation were,retrospectively analyzed.ResultsAll patients with acute encephalocele were associated with delayed intracranial hematoma.The effective treatment of acute encephalocele was to remove the hematoma out of the brain timely and thoroughly.33 patients survived the disorder of acute encephalocele,of which,good recovery occurred in 20 patients,moderate disability in 6 patients,severe disability in 4 patients,and vegetative state in 3 patients according to Glasgow Outcome Score 3 months later.ConclusionHead-CT scanning should be timely performed when intracperative acute encephalocele occurred,whereas the measures of efficacy improvement on acute encephalocele were to timely.