In 2025， Severe Liver Disease and Artificial Liver Group and Nutrition and Regeneration in End-Stage Liver Disease Group of Chinese Society of Hepatology， Chinese Medical Association， convened a panel of national experts to jointly develop China’s first guideline for the diagnosis and treatment of acute-on-chronic liver failure （ACLF）. Based on the latest research findings and clinical practice in China and globally， this guideline establishes a standardized definition of ACLF and provide recommendations for its diagnosis， treatment， and clinical management. This article gives an interpretation of the key points in the guideline， in order to provide a reference for standardized diagnosis and treatment of ACLF.

Objective To investigate the physiological characteristics of Bacillus thuringiensis subspecies israelensis (Bti) with double mutations of cwlE and sigK genes and to assess the activity of Bti against larvae of Culex pipiens pallens under different external factors, so as to provide the theoretical evidence for the use of engineered bacteria of Bti for effective mosquito control. Methods B. thuringiensis wild-type strain Bt-59 and Bt-59 strain with cwlE mutation [Bt-59 (ΔcwlE)] were cultured in nutrient broth media for 24 hours, and Bt-59 strains with sigK mutation [Bt-59 (ΔsigK)] and double mutations of cwlE and sigK [Bt-59 (ΔcwlE-sigK)] were cultured in nutrient broth media for 48 hours. Then, 5 μL of culture media were transferred to glass sides, and cell morphology and mother cell lysis were observed under an optical microscope. The optical densities of Bti strain culture media were measured at different time points of culture, and the growth curves of Bt-59, Bt-59 (ΔcwlE), Bt-59 (ΔsigK), and Bt-59 (ΔcwlE-sigK) strains were plotted. The differences in carbon source metabolism of four Bti strains were analyzed using the Biolog microplate culture method, and the metabolic activity of these strains was estimated with average well color development (AWCD). The fermentation media of these four Bti strains were diluted into final concentrations of 2.000, 1.000, 0.500, 0.250, and 0.125 μL/L, and the median lethal concentrations (LC₅₀ values) of these four strains against the third instar larvae of Cx. pipiens pallens were determined. In addition, the fermentation media of Bti strains were processed as follows: pH adjusted to 5, 7 and 9; treated at 30, 40 ℃ and 50 ℃ for 12 hours; and exposed to irradiation with ultraviolet lights for 0 hour and 6 hours. Then, 20 third instar larvae of Cx. pipiens pallens were exposed to the above processed fermentation media at a final concentration of 1 μL/L in 200 mL of water at 26 ℃ for 24 hours, and the mosquito mortality was estimated to evaluate the effects of pH, temperature and ultraviolet irradiation on the larvicidal activity of four Bti strains. Results The growth curves of the Bt-59 strain and its mutants shared a similar changing trend, and both experienced a stable phase 6 hours post-culture. Both spores and crystal proteins were found in Bt-59 and Bt-59 (ΔcwlE) cells, and only crystal proteins were found in Bt-59 (ΔsigK) and Bt-59 (ΔcwlE-sigK) cells. No lysis was found in the cell wall of the Bt-59 (ΔcwlE-sigK) strain, and the crystal protein was embedded in the mother cell. Biolog microplate culture assay showed that the AWCD values of four Bti strains showed a similar changing trend over time, and 33 carbon sources were found to be metabolized by all of the four strains, including dextrin, D-maltose and D-trehalose. The LC50 values of the fermentation media of Bt-59, Bt-59 (ΔcwlE), Bt-59 (ΔsigK), and Bt-59 (ΔcwlE-sigK) strains were 0.60, 0.51, 0.70 μL/L and 0.72 μL/L against Cx. pipiens pallens, respectively. The adjusted mortality of larval Cx. pipiens pallens reduced by 76.60%, 76.00%, 66.67%, and 0 following exposure to the fermentation media of Bt-59, Bt-59 (ΔcwlE), Bt-59 (ΔsigK), and Bt-59 (ΔcwlE-sigK) strains at a pH of 5 relative to at a pH of 7, and the adjusted mortality reduced by 49.02%, 51.06%, 36.36%, and 4.44% following 6-hour exposure to ultraviolet irradiation relative to 0-hour exposure, while the adjusted mortality was 68.33% to 83.33% following treatment with the fermentation media of four Bti strains at different temperatures. Conclusions Bt-59 (ΔcwlE-sigK) strains do not generate spores, and the absence of cwlE and sigK does not affect the growth, carbon source metabolism, and larvicidal activity of Bti strains against larval Cx. pipiens pallens. Cell wall embedding of Bt-59 (ΔcwlE-sigK) strains may protect larvicidal crystal proteins of Bti strains from external environmental factors, including ultraviolet irradiation, and pH alteration.

Objective:To investigate the current situation of fall fear in elderly patients after operation of intertrochanteric fracture, analyze the influencing factors and build a prediction model.Methods:A prospective cohort study and convenient sampling method were used to select 303 elderly patients with intertrochanteric fracture after surgery in the Department of Orthopedics, Affiliated Hospital of Jining Medical University from January 2021 to December 2022 as the modeling group. According to the scores of fall fear when getting out of bed for the first time, they were divided into 211 cases in the fall fear group and 92 cases in the non-fall fear group. Multivariate Logistic regression was used to analyze the influencing factors of fall fear after operation of femoral intertrochanteric fracture in elderly patients, and a risk prediction score model was established. ROC curve was used to evaluate the differentiation of the model, calibration chart was used to evaluate the calibration degree of the model, and Bootstrap method was used for internal verification. Data of 81 elderly patients with intertrochanteric fracture treated from January to May 2023 were collected as a validation set for external verification of the model.Results:A total of 6 influencing factors were included in the prediction of fall fear risk model for elderly patients after operation of intertrochanteric fracture: gender ( OR = 3.229, 95% CI 1.672 - 6.401), number of co-existing diseases ( OR = 9.578, 95% CI 4.532 - 20.245), pain ( OR = 1.684, 95% CI 1.074 - 2.463), depression ( OR = 1.719, 95% CI 1.371 - 2.155), social support ( OR = 0.859, 95% CI 0.784 - 0.942), and ability to perform activities of daily living ( OR = 0.960, 95% CI 0.932 - 0.989) (all P<0.05). The area under ROC curve of the prediction model was 0.853, the sensitivity was 0.896, and the specificity was 0.663. The area under the externally verified ROC curve was 0.766, the sensitivity was 0.800, and the specificity was 0.645. The calibration curve was close to the reference curve in the model prediction, indicating that the model hasd a good prediction effect. Conclusions:The prediction effect of this model is good, and it can quickly predict the risk of fall fear in elderly patients after operation of intertrochanteric fracture, and provide a basis for medical personnel to provide timely intervention measures.

ObjectiveTo investigate the value of MELD 3.0， MELD， and MELD-Na scores in assessing the 90-day prognosis of patients with acute-on-chronic liver failure （ACLF） through a comparative study. MethodsA retrospective analysis was performed for the clinical data of 605 patients with ACLF who were treated in Tianjin Third Central Hospital， The Fifth Medical Center of Chinese PLA General Hospital， and Beijing YouAn Hospital from November 2012 to June 2019， and according to the 90-day follow-up results after admission， they were divided into survival group with 392 patients and death group with 213 patients. The receiver operating characteristic （ROC） curve， the area under the ROC curve （AUC）， net reclassification improvement （NRI）， integrated discrimination improvement （IDI）， and decision curve analysis （DCA） curve were used to investigate the value of MELD 3.0， MELD， and MELD-Na scores at baseline， day 3， week 1， and week 2 in predicting the prognosis of the disease. ResultsAt day 3 and week 1， MELD 3.0 score had an AUC of 0.775 and 0.808， respectively， with a better AUC than MELD score （P<0.05）. At day 3， week 1， and week 2， MELD 3.0 score showed an NRI of 0.125， 0.100， and 0.081， respectively， compared with MELD in predicting the prognosis of ACLF patients， as well as an NRI of 0.093， 0.140， and 0.204， respectively， compared with MELD-Na score in predicting prognosis. At baseline， day 3， week 1， and week 2， MELD 3.0 showed an IDI of 0.011， 0.025， 0.017， and 0.013， respectively， compared with MELD in predicting the prognosis of ACLF patients. At day 3 and week 2， MELD 3.0 showed an IDI of 0.027 and 0.038， respectively， compared with MELD-Na in predicting the prognosis of ACLF patients. All the above NRIs and IDIs were >0， indicating a positive improvement （all P<0.05）. DCA curves showed that MELD 3.0 was superior to MELD at day 3 and was significantly superior to MELD-Na at week 2. There was no significant difference in the ability of the three scores in predicting the prognosis of ACLF patients with different types， and there was also no significant difference in the ability of the three scores in predicting the prognosis of ACLF patients with the etiology of HBV infection， alcohol， or HBV infection combined with alcohol， while MELD 3.0 was superior to MELD for ACLF patients with other etiologies （P<0.05）. ConclusionMELD 3.0 score is better than MELD and MELD-Na scores in predicting the 90-day survival of patients with ACLF， but with limited superiority.

Background::Sirtuin-3 (Sirt3) has been documented to protect against mitochondrial dysfunction and apoptosis. Honokiol (HKL) is a Sirt3 pharmacological activator with reported neuroprotective effects in multiple neurological disorders. The present study aimed to explore the neuroprotective effects of HKL and the role of Sirt3 following intracerebral hemorrhage (ICH).Methods::An in vivo ICH model in rats was established by injecting autologous blood into the right basal ganglia. PC12 cells were stimulated with hemin. For the in vivo investigation, the modified Neurological Severity Scores and the Morris water maze test were performed to assess neurological deficits. Hematoxylin-Eosin and Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining were employed to evaluate the histopathology and apoptosis. Immunohistochemical staining was used to investigate the expression of Sirt3. Adenosine triphosphate (ATP) levels were quantified to assess mitochondrial dysfunction. Cell counting kit-8, lactate dehydrogenase assay, and flow cytometry were used to analyze cell vitality and apoptosis in vitro. Immunofluorescence staining was performed to observe mitochondrial morphology and dynamin-related protein 1 (Drp1) localization to mitochondria. Western blot was applied to quantify the expression of Sirt3, Bax, Bcl-2, cleaved-caspase-3, Drp1, phosphorylation of Drp1 at serine-616, and phosphorylation of Drp1 at serine-637 in vivo and in vitro.Results::HKL treatment alleviated neurological deficits, attenuated the histopathological damage and cell apoptosis, and restored the decreased ATP levels in ICH rats. HKL improved cell survival rate, reduced cell apoptosis, and inhibited mitochondrial fission in PC12 cells. Moreover, both in vivo and in vitro models showed increased phosphorylation of Drp1 at Ser616, and reduced phosphorylation of Drp1 at Ser637. Meanwhile, immunofluorescence co-localization analysis revealed that hemin increased the overlap of Drp1 and mitochondria in PC12 cells. The phosphorylation and mitochondrial translocation of Drp1 were effectively reversed by HKL treatment. Importantly, the selective Sirt3 inhibitor 3-(1H-1,2,3-triazol-4-yl) pyridine suppressed these effects. Conclusion::Our findings demonstrated that HKL ameliorated ICH-induced apoptosis and mitochondrial fission by Sirt3, suggesting that HKL has immense prospects for the treatment of ICH.

Objective:To analyze the clinical characteristics among types of acute-on-chronic liver failure (ACLF) and explore the new classification criteria for judging the prognosis of acute-on-chronic liver failure, so as to provide a basis for the formulation of more precise therapeutic schedule.Methods:388 cases with ACLF diagnosed in two tertiary level hospitals were included. Patients demographic characteristics, clinical examination information, diagnostic and treatment process information were collected. Laboratory examination data of day 1, 3, 7, 14, 21, 28 and of week 12 or prior to discharge after improvement and at 24 h prior to liver transplantation or death from the diagnosis of ACLF were collected. According to the change trend of the patient's prothrombin activity (PTA), the changes within 4 weeks and 12 weeks were divided into: increased to > 40 %, increase but still ≤ 40%, progressively decreasing or not continuously rising. Moreover, the change trend of total bilirubin (TBil) was divided into: decreasing degree≥50%, decreasing degree < 50%, progressively increasing or not decreasing. Patients meeting the requirements of dynamic classification were screened. PTA and TBil variation tendency of each patient at week 4 and 12 was synthesized, and prognostic condition for dynamic classification was formulated. The clinical characteristics of ACLF patients were analyzed by χ2 test. Results:A total of 262 screened cases were enrolled. At the 4th week of the course of disease, 45% of the patients' PTA had increased to > 40%, and 40.8% of the patients' TBIL had decreased by 50%. When the course of disease was progressed to 12 weeks, 65.3% of the patients' PTA had increased to > 40%, and 63.4% of the patients' TBIL had decreased by 50%. Combined with the prognosis of the patients at the 4th and 12th week, the patients' disease evolution process was divided into five types: Type A: 60 cases (22.9%) of rapid progression; Type B: 82 cases (31.3%) of rapid recovery; Type C: 48 cases (18.3%) of slow progression; Type D: 43 cases (16.4%) of slow recovering; Type E: 29 cases (11.1%) of slow persistence. The proportions of patients with rapid progression combined with upper gastrointestinal hemorrhage, hepatic encephalopathy, and acute renal injury were 16.7%, 33.3%, and 33.3%, respectively; while the above-mentioned complications accounted for 3.7%, 7.3%, and 12.2% only in the rapid recovery type, χ2 = 14.411, 20.060, 12.140, P < 0.05, and the differences were statistically significant. Fungal infection rates were 21.7%, and 10.4% in patients who died of disease or liver transplantation (i.e., patients with rapid progression and slow-progressing types), respectively, and 1.2%, 14%, and 6.9% in patients with rapid progression type, slow-recovering type, and slow persistence type, respectively, and the difference between the rapid progression type and the rapid recovery type was significant, χ2 = 18.925, and the difference was statistically significant ( P < 0.05). Conclusion:The course of disease progression in ACLF patients can be divided into rapid progression type, rapid recovery type, slow progression type, slow recovering type, and slow persistence type. The basis of liver disease, accompanied with fungal infection, gastrointestinal hemorrhage, hepatic encephalopathy and acute renal injury can affect the development of ACLF.

Objective:To compare the clinical features between patients with acute-on-chronic liver failure (ACLF) and decompensated liver cirrhosis (DC) combined with acute kidney injury (AKI).Methods:Demographic data, clinical examination results, diagnosis and treatment information of ACLF and DC patients were collected retrospectively. Clinical characteristics of ACLF combined with AKI and DC combined with AKI and their impact on the 90-day mortality risk were compared.Results:The clinical characteristics of patients with ACLF-AKI and DC-AKI were compared. The results showed that the leukocyte count, absolute neutrophil count, alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil) of ACLF-AKI patients were higher than those of DC-AKI patients, while prothrombin activity (PTA), and albumin were lower than those of DC-AKI patients, and the difference was statistically significant ( P < 0.05). The co-infection rate in patients with ACLF-AKI was significantly higher than that of DC-AKI group (96.9% vs. 39.5%) ( P < 0.05), and during the diagnosis of AKI, the median value of serum creatinine in ACLF patients was 147 μmol / L (IQR: 122-189), while that in DC group was 123.5 μmol / L (IQR: 103.8-155.5), and the difference between the two groups was statistically significant ( P < 0.05). According to the HRS-AKI diagnostic criteria for liver cirrhosis, 44 (68.8%) cases of ACLF-AKI met the diagnosis of HRS -AKI, which was significantly higher than the proportion of 18 (47.4%) cases of DC-AKI ( P < 0.05). Four (10.5%) cases of DC-AKI had died or underwent liver transplantation within 30 days and eight (21.1%) cases had died or underwent liver transplantation within 90 days, while 22 (34.4%) cases of ACLF-AKI patients had died or underwent liver transplantation within 30 days and 35 (54.7%) cases had died or underwent liver transplantation within 90 days, and χ2 values was 7.140 and 11.062, respectively ( P < 0.05). The results of multivariate regression analysis suggested that the independent risk factors that affect the 90-days mortality rate of DC patients were hepatic encephalopathy, gastrointestinal bleeding, and TBil, while the independent risk factors affecting the 90-days death risk of ACLF patients included AKI, PTA and TBil. Conclusion:Compared with DC-AKI patients, ACLF-AKI patients have a higher proportion of infection rate, higher serum creatinine level when diagnosed AKI, and faster disease progression, leading to a greater risk of death.

Objective To explore the changes of Beclin-1,P62/SQSTM1,microtubule-associated protein 1 light chain 3 (LC3)and unc-51 like autophagy activating kinase 1 (ULK-1)in the brains of the rats in the deve-lopmental stage with epilepsy. Methods Seventy-two male Sprague Dawley (SD)rats aged 21 days were randomly divided into the control group and the epilepsy group. The rats in 2 groups were randomly subdivided into 4 groups according to the time intervals (3 h,6 h,12 h and 48 h),respectively,with 9 rats in each group. The rats in the epilep-sy group were injected with kainic acid (12 mg/kg)to induce epilepsy,and the rats in the control group were injected with equal volume of saline. The rats in 2 groups were anaesthetized and sacrificed. Then,the brain tissues of the rats were quickly removed according to the time intervals. The brain damages were determined by adopting Nissl staining method. The apoptotic cells were detected by Terminal - deoxynucleoitidyl transferase mediated nick end labeling (TUNEL)assays. The expressions of Beclin-1,P62/SQSTM1,LC3 and ULK-1 mRNA levels in cortex were mea-sured by using real-time quantitative polymerase chain reaction (qPCR)analysis. Results Nissl staining indicated that many neurons were damaged performing vague outline,irregularly aligned,pyknotic nuclei and shrunken somata in the epilepsy 48 h group. In addition,there was a huge loss of neurons in cortex in the epilepsy 48 h group [(82 ± 8)num-bers],compared with the control group [(122 ± 8)numbers],and the difference was statistically significant (F＝3. 768, P＝0. 01). The apoptotic cells tremendously increased in the epilepsy 48 h group [(13 ± 7)numbers],compared with the control group [(2 ± 1)numbers]by TUNEL analysis,and the diffe-rence was statistically significant (t＝ -3. 821, P＝0. 003). qPCR showed the mRNA levels of Beclin-1,P62/SQSTM1,LC3 and ULK-1 were upregulated in the epi-lepsy 12 h group (1. 70 ± 0. 75,1. 75 ± 0. 77,1. 52 ± 0. 43,7. 48 ± 6. 12)and the epilepsy 48 h group (1. 63 ± 0. 43, 1. 48 ± 0. 74,1. 74 ± 0. 55,7. 69 ± 5. 65),compared with the control group (1. 00,1. 00,1. 00,1. 00),and the differences were statistically significant (F＝2. 820,3. 452,5. 811,5. 002,all P＜0. 05). Conclusion The autophagy activates be-fore apoptosis occurs,and autophagy-related genes probably are involved in epilepsy-induced brain damage.

Objective:To compare the cognitive functions in patients with depressive disorder,hyperlipidemia disorder,and comorbid both of the disorders.Methods:A cross-sectional study was performed in age,gender and education year matched patients with depressive disorder (n =51)(according to the ICD-10),hypedipidemia(n =38) (according to the Chinese adult lipid guideline),comorbid both of the disorders(n =40) and normal controls (n =56) were recruited in this study.All subjects received a battery of neuropsychological tests to access the anxiety and depression symptoms and cognitive function.Results:The scores of MoCA were lower in the patients with comorbid both disorders and patients with depression than patients with hypedipidemia [(24 ± 3),(24 ± 4)vs.(26 ± 3),Ps ＜0.05],and were lower in patients with depression than in normal controls(25 ±3),P ＜0.05.Stroop color test amends numbers were higher in patients with comorbid disorder than in the other three groups (Ps ＜0.05).The scores of immediate and delayed logical memory were higher in patients with hyperlipidemia than in other three groups (Ps ＜0.05).The total number of words in verbal fluency test were lower in patients with comorbid disorders and patients with depression than in patients with hyperlipidemia (Ps ＜0.05).Wisconsin card sorting test category completes were lower in patients with comorbid disorders and patients with depression than in patients with hyperlipidemia and normal controls (Ps ＜ 0.05).The scores of persistent errors were higher in patients with comorbid disorders and patients with depression than in patients with hyperlipidemia and normal controls (Ps ＜0.05).Conclusion:In this study,patients with depressive disorder have impairment of cognitive function,while hyperlipidemia may probably do not impair cognitive function.

BACKGROUND:The application of virtual reality technology in preoperative simulation can reduce the risk in operation effectively and improve the quality of surgery. Virtual reality technology has very important practical significance in the application of surgery. OBJECTIVE:The reverse engineering and virtual reality technology were used to achieve the preoperative simulation of a case of Pilon fracture. METHODS:The affected bones were reconstructed according to the CT data using the patient’s ankle portion in MIMICS software, and the separated bone fractures were restored. According to the characteristics of virtual reality technology, further processing on bone model was carried out using Geomagic software;the models of the fractured bones were exported with STL format. The restored bone fragments were checked up to determine integrity in the virtual reality operation platform. These models were used to do simulated operations in the virtual reality operation platform. RESULTS AND CONCLUSION:A case of Pilon fracture was simulated by using reverse engineering and virtual reality technology preoperatively. The processed 3D model was introduced into the virtual reality system to simulate the operation and help doctors choose the type, position and direction of the surgical approach and plate;the effect is good.