Objective:To explore the therapeutic effect of ultrasound intervention combined with support in the treatment of diabetic patients with severe cholecystitis and the risk factors of septic shock.Methods:A total of 81 diabetic patients with severe cholecystitis treated in the emergency department of Beijing Friendship Hospital from January to December 2021 were retrospectively selected and divided into sepsis group ( n=41) and septic shock group ( n=40). The clinical data of the patients were collected, and the curative effect of ultrasound intervention combined with supportive treatment for diabetic patients with severe cholecystitis was analyzed. logistic regression and receiver operating characteristic (ROC) curve were used to predict the risk factors for diabetic patients with severe cholecystitis to develop septic shock. Results:In the sepsis group, 5 cases were positive in bile culture and 2 cases were positive in blood culture. In the septic shock group, 28 cases were positive in bile culture and 12 cases were positive in blood culture. The concentration of glycosylated hemoglobin (HbA 1c) before treatment, gallbladder width and final 28-day all-cause mortality in the sepsis group were lower than those in the shock group (all P<0.05). Before treatment, procalcitonin (PCT), lactic acid (Lac), shock index (SI) and Sequential Organ Failure Assessment (SOFA) scores in the sepsis group were lower than those in the sepsis shock group (all P<0.05). The mean arterial pressure (MAP) was higher than that of the shock group ( P<0.01). After treatment for the sepsis group and the septic shock group, white blood cell (WBC), neutrophilic granulocyte percentage (NEU%), platelet (PLT), PCT, C-reactive protein (CRP), total bilirubin (TBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST), MAP, heart rate (HR), Lac, SI and SOFA scores were improved (all P<0.05). After treatment, D-dimer decreased in the sepsis group ( P<0.05), but there was no significant difference in D-dimer between the sepsis shock group and before treatment ( P=0.729 5). Multivariate logistic regression showed that HbA 1c, PCT and MAP were independent risk factors for septic shock in diabetic patients with severe cholecystosis ( OR=9.19, 1.32, 0.58, all P<0.05). The area under ROC curve of SOFA score, HbA 1c and PCT for predicting septic shock due to cholecystitis in diabetic patients were 0.878, 0.918 and 0.715. Conclusions:Ultrasound intervention combined with supportive treatment can significantly alleviate the condition of patients with severe cholecystitis, but early intervention is still needed to reduce the risk of death. HbA 1c and PCT can be used as independent risk factors for septic shock in diabetic patients with severe cholecystitis.

Background::The ability to generate functional hepatocytes without relying on donor liver organs holds significant therapeutic promise in the fields of regenerative medicine and potential liver disease treatments. Clustered regularly interspaced short palindromic repeats (CRISPR) activator (CRISPRa) is a powerful tool that can conveniently and efficiently activate the expression of multiple endogenous genes simultaneously, providing a new strategy for cell fate determination. The main purpose of this study is to explore the feasibility of applying CRISPRa for hepatocyte reprogramming and its application in the treatment of mouse liver fibrosis.Method::The differentiation of mouse embryonic fibroblasts (MEFs) into functional induced hepatocyte-like cells (iHeps) was achieved by utilizing the CRISPRa synergistic activation mediator (SAM) system, which drove the combined expression of three endogenous transcription factors— Gata4, Foxa3, and Hnf1a—or alternatively, the expression of two transcription factors, Gata4 and Foxa3. In vivo, we injected adeno-associated virus serotype 6 (AAV6) carrying the CRISPRa SAM system into liver fibrotic Col1a1-Cre ER; Cas9 fl/fl mice, effectively activating the expression of endogenous Gata4 and Foxa3 in fibroblasts. The endogenous transcriptional activation of genes was confirmed using real-time quantitative polymerase chain reaction (RT-qPCR) and RNA-seq, and the morphology and characteristics of the induced hepatocytes were observed through microscopy. The level of hepatocyte reprogramming in vivo is detected by immunofluorescence staining, while the improvement of liver fibrosis is evaluated through Sirius red staining, alpha-smooth muscle actin (α-SMA) immunofluorescence staining, and blood alanine aminotransferase (ALT) examination. Results::Activation of only two factors, Gata4 and Foxa3, via CRISPRa was sufficient to successfully induce the transformation of MEFs into iHeps. These iHeps could be expanded in vitro and displayed functional characteristics similar to those of mature hepatocytes, such as drug metabolism and glycogen storage. Additionally, AAV6-based delivery of the CRISPRa SAM system effectively induced the hepatic reprogramming from fibroblasts in mice with live fibrosis. After 8 weeks of induction, the reprogrammed hepatocytes comprised 0.87% of the total hepatocyte population in the mice, significantly reducing liver fibrosis. Conclusion::CRISPRa-induced hepatocyte reprogramming may be a promising strategy for generating functional hepatocytes and treating liver fibrosis caused by hepatic diseases.