Non-alcoholic fatty liver disease （NAFLD） is a chronic liver disease closely related to metabolism， which is mainly characterized by abnormal lipid deposition in hepatocytes. In recent years， with the increasing prevalence of obesity and metabolic syndrome， NAFLD has become one of the most common chronic diseases in the world. The pathogenesis of NAFLD is complex and varied， involving the cross-regulation of multiple signaling pathways such as glucose-lipid metabolism， oxidative stress， and inflammation. The TLR4 signaling pathway plays a key role in the development and progression of NAFLD， and abnormal activation of this pathway accelerates the deterioration of NAFLD by promoting the release of pro-inflammatory cytokines， inducing oxidative stress， and exacerbating insulin resistance. Studies have shown that traditional Chinese medicine （TCM） can regulate the TLR4 signaling pathway to alleviate the symptoms and pathological features of NAFLD. The present review summarizes the experimental research progress in the TCM regulation of the TLR4 signaling pathway in treating NAFLD in the past 5 years， covering a wide range of TCM active ingredients （such as polysaccharides， terpenoids， alkaloids， flavonoids） and compound prescriptions. The active ingredients and compound prescriptions of TCM can effectively ameliorate lipid metabolism disorders， reduce insulin resistance， regulate intestinal flora， and inhibit inflammation and oxidative stress by regulating the TLR4 signaling pathway via multiple targets and pathways， thus slowing down the progression of NAFLD. Through in-depth analysis of the pathological mechanisms of NAFLD and exploration of the potential of TLR4 signaling pathway as a therapeutic target， we can provide theoretical support for the application of TCM in the treatment of NAFLD， as well as new perspectives and directions for future clinical research and new drug development， thereby promoting the innovation and development of therapeutic strategies for NAFLD.

Currently,human health due to corona virus disease 2019(COVID-19)pandemic has been seriously threatened.The coronavirus severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)spike(S)protein plays a crucial role in virus transmission and several S-based therapeutic approaches have been approved for the treatment of COVID-19.However,the efficacy is compromised by the SARS-CoV-2 evolvement and mutation.Here we report the SARS-CoV-2 S protein receptor-binding domain(RBD)inhibitor licorice-saponin A3(A3)could widely inhibit RBD of SARS-CoV-2 variants,including Beta,Delta,and Omicron BA.1,XBB and BQ1.1.Furthermore,A3 could potently inhibit SARS-CoV-2 Omicron virus in Vero E6 cells,with EC50 of 1.016 pM.The mechanism was related to binding with Y453 of RBD deter-mined by hydrogen-deuterium exchange mass spectrometry(HDX-MS)analysis combined with quan-tum mechanics/molecular mechanics(QM/MM)simulations.Interestingly,phosphoproteomics analysis and multi fluorescent immunohistochemistry(mIHC)respectively indicated that A3 also inhibits host inflammation by directly modulating the JNK and p38 mitogen-activated protein kinase(MAPK)path-ways and rebalancing the corresponding immune dysregulation.This work supports A3 as a promising broad-spectrum small molecule drug candidate for COVID-19.

Objective This study investigated the impact of occupational mercury(Hg)exposure on human gene transcription and expression,and its potential biological mechanisms. Methods Differentially expressed genes related to Hg exposure were identified and validated using gene expression microarray analysis and extended validation.Hg-exposed cell models and PTEN low-expression models were established in vitro using 293T cells.PTEN gene expression was assessed using qRT-PCR,and Western blotting was used to measure PTEN,AKT,and PI3K protein levels.IL-6 expression was determined by ELISA. Results Combined findings from gene expression microarray analysis,bioinformatics,and population expansion validation indicated significant downregulation of the PTEN gene in the high-concentration Hg exposure group.In the Hg-exposed cell model(25 and 10 μmol/L),a significant decrease in PTEN expression was observed,accompanied by a significant increase in PI3K,AKT,and IL-6 expression.Similarly,a low-expression cell model demonstrated that PTEN gene knockdown led to a significant decrease in PTEN protein expression and a substantial increase in PI3K,AKT,and IL-6 levels. Conclusion This is the first study to report that Hg exposure downregulates the PTEN gene,activates the PI3K/AKT regulatory pathway,and increases the expression of inflammatory factors,ultimately resulting in kidney inflammation.

Objective To establish a nomogram model for efficiently predicting overall survival(OS)and cancer-specific survival(CSS)in patients with CRCHBM.Method 2239 patients from 2010 to 2019 were retrospectively analyzed from the Surveillance,Epidemiology,and End Results Program(SEER)databases and Wuhan Union Hospital Cancer Center.SEER is randomly assigned to the training and internal validation cohorts,and the Wuhan database serves as the external validation.Cox regression analyses were used to determine the independent clinicopathological prognosis factors affecting OS and CSS,and a nomogram was constructed to predict OS and CSS.The clinical utility of columnar plots was assessed using calibration curves,area under the curve(AUC),and decision curve analysis(DCA).Result OS column line graphs were constructed based on nine independent predictors:age,tumor location,degree of differentiation,tumor size,TNM stage,chemotherapy,primary focus surgery,number of lymph nodes sampled,and serum carcinoembryonic antigen(CEA)level.The C-index of the nomogram to predict the 1-,3-,and 5-year OS were 0.764,0.790,and 0.805 in the training group,0.754,0.760,and 0.801 in the internal validation group,and 0.822,0.874,and 0.906 in the external validation group.CSS column line graphs were constructed based on 3 independent predictors of TNM staging,radiotherapy and chemotherapy.The 1-,3-,and 5-year CSS AUROC values of the training group were 0.791,0.757,and 0.782,respectively.0.682,0.709,0.625 in the internal validation group and 0.759,0.702,0.755 in the external validation group,respectively.The results of receiver operating characteristic curve(ROC),ROC and DCA showed that the use of our model was more effective in predicting OS and CSS than other single clinicopathological features.Conclusion In summary,the nomogram based on significant clinicopathological features can be conveniently used to predict OS and CSS individually in patients with CRCHBM.

Objective To explore the treatment for endometriosis(EMs)by Professor SITU Yi,a Guangdong famous traditional Chinese medicine physician,thus to provide reference for the diagnosis and treatment of EMs in traditional Chinese medicine based on personalized knowledge graph analysis and data mining technology.Methods Based on the literature research achievements of diagnosis and treatment of EMs,the ontology knowledge tree of famous physician's principles,methods,prescriptions and medicines was established by user-computer integration method.The medical records of EMs patients treated by Professor SITU Yi in the outpatient department of Guangdong Provincial Hospital of Chinese Medicine from 2013 to 2019 were collected.After standardization of the data of medical records,the knowledge set was defined,the knowledge map of TCM diagnosis and treatment of EMs was constructed,and the symptom characteristics and medication rules for EMs were analyzed.Results The core symptoms of EMs patients were dysmenorrhea,chronic pelvic pain and infertility,with the predominated syndrome of blood stasis.The pathogenesis of EMs patients was mainly due to qi stagnation,kidney deficiency,and qi deficiency.Frequent itemset analysis yielded the core drug groups of"Corydalis Rhizoma,Cyperi Rhizoma,Paeoniae Radix Alba""Lycii Fructus,Taxilli Herba,Poria,Dioscoreae Rhizoma"and"Aurantii Fructus,Leonuri Herba".The EMs word cloud map constructed on the basis of knowledge element clustering and neural network algorithm showed that the drug combinations of"Salviae Miltiorrhizae Radix et Rhizoma,Paeoniae Radix Rubra""Corydalis Rhizoma,Curcumae Radix""Cinnamomi Ramulus,Moutan Cortex,Paeoniae Radix Rubra,Poria""Salviae Miltiorrhizae Radix et Rhizoma,Paeoniae Radix Rubra,Sparganii Rhizoma,Curcumae Rhizoma""Chuanxiong Rhizoma,Corydalis Rhizoma"and"Taxilli Herba,Cuscutae Semen"were commonly used for the treatment of EMs by Professor SITU Yi.The results of hierarchical clustering analysis and association network construction for Chinese medicinals,symptoms and syndromes showed that there were complex inner link among the syndromes,symptoms and medication.Conclusion For the treatment of EMs,Professor SITU Yi usually adopt the methods of activating blood to remove stasis and tonifying kidney,and also performs the periodic therapy and simultaneous application of purging and nourishing therapeutics.Knowledge graph analysis and data mining based on semantic web are helpful to unveil the tacit knowledge of clinical diagnosis and treatment experience of Professor SITU Yi,which can provide reference for the differentiation and treatment of EMs with Chinese medicine.

Objective:To explore the role and mechanism of P311 in the differentiation of mouse skin fibroblasts (Fbs) into myofibroblasts.Methods:The study was an experimental research. Six 2-day-old male C57BL/6 mouse were used to extract skin Fbs by enzymatic hydrolysis method and routinely cultured. The 1 st to 3 rd passage cells were taken and divided into empty vector group transfected with empty adenovirus and P311 group transfected with P311 high expression adenovirus, and P311+myocardin-related transcription factor A (MRTF-A) small interfering RNA (siMRTF-A) group transfected with P311 high expression adenovirus and siMRTF-A according to the random number table. After 72 h of culture, the cell proliferation vitality of cells in 3 groups was detected by cell counting kit 8, the protein expressions of MRTF-A, α-smooth muscle actin (α-SMA), and serum response factor (SRF) in cells in 3 groups were detected by Western blotting, the collagen gel contraction assay was performed and the 72 h gel contraction rates in 3 groups were calculated. The sample numbers in the above experiments were all 3. The protein expressions of MRTF-A and SRF in cells, cytoplasm, and nucleus in cells in empty vector group and P311 group were detected by Western blotting, with sample number of 4. Results:After 72 h of culture, the cell proliferation vitality of cells in empty vector group, P311 group, and P311+siMRTF-A group was similar ( P>0.05). After 72 h of culture, compared with those in empty vector group, the protein expressions of MRTF-A, α-SMA, and SRF in cells in P311 group were significantly increased ( P<0.05), while the protein expressions of MRTF-A and SRF in cells in P311+siMRTF-A group were significantly decreased ( P<0.05). Compared with those in P311 group, the protein expressions of MRTF-A, SRF, and α-SMA in cells in P311+siMRTF-A group were significantly decreased ( P<0.05). The 72 h gel contraction rate showing cell contractility in P311 group was (84.8±6.2)%, which was significantly higher than (27.8±2.6)% in empty vector group and (24.7±3.2)% in P311+siMRTF-A group (with P values all <0.05). The 72 h gel contraction rates in empty vector group and P311+siMRTF-A group were similar ( P>0.05). After 72 hours of culture, the protein expressions of MRTF-A (with t values of 5.86 and 3.77, respectively, P<0.05) and SRF (with t values of 3.95 and 3.97, respectively, P<0.05) in cells and cytoplasm in P311 group were significantly higher than those in empty vector group, while the protein expressions of MRTF-A and SRF in the nucleus of cells were similar between the two groups ( P>0.05). Conclusions:P311 can promote the differentiation of fibroblasts into myofibroblasts through MRTF-A, and then participate in scar formation.

Objective To investigate the effect of hairy and enhancer of split 5(HES5)on transdifferentiation and apoptosis of renal tubular epithelial cells induced by TGF-β1 and its potential mechanism.Methods The differentially expressed genes in GSE66494 data were analyzed and screened.The mouse model of unilateral uretera obstruction(UUO)was established,and the expression level of HES5 was detected in the renal tissue.HK-2 cells were treated with 10 ng/mL TGF-β1 for 24 h to establish a tubular epithelial-mesenchymal transition(EMT)model,and then qRT-PCR and Western blotting were performed to detect the expression of HES5 at mRNA and protein levels.After HK-2 cells were transfected with the plasmid overexpressing HES5,the protein levels of fibronectin,collagen Ⅰ,vimentin,apoptosis markers Bax and Bcl2 were detected in 24 h later.Then,HK-2 cells were divided into Control group,siHES5 group,TGF-β 1 group,and siHES5+TGF-β1 group.The protein level of fibrosis and apoptosis markers were measured in above groups with Western blotting.TUNEL staining and flow cytometry were employed to detect cell apoptosis.Western blotting was applied to determine the protein levels of AKT,p-AKT,PI3K and p-PI3K.HK-2 cells overexpressing HES5 were treated with PI3K inhibitor LY294002,and the expression of vimentin was detected.Results The expression of HES5 was significantly up-regulated in both chronic kidney disease(CKD)and fibrotic kidneys of mice.Overexpression of HES5 promoted the synthesis of fibronectin,collagen Ⅰ,vimentin and Bax in HK-2 cells,and inhibited the expression of Bcl2(P＜0.05).HES5 knockdown not only down-regulated the expression of fibrosis markers,but also inhibited the apoptosis of HK-2 cells.Furthermore,HES5 knockdown inhibited the activation of PI3K/AKT signaling pathway induced by TGF-β1 in HK-2 cells(P＜0.05).Inhibitors of the PI3K/AKT signaling pathway inhibitor attenuated the induction of HES5 on vimentin.Conclusion HES5 knockdown inhibits the transdifferentiation and apoptosis in TGF-β1-induced renal tubular epithelial cells,which may be related to the decreased activity of the PI3K/AKT signaling pathway.

Objective:To preliminarily evaluate the effects and analyze the influencing factors of continuous renal replacement therapy (CRRT) in severe burn patients complicated with acute kidney injury (AKI).Methods:This study was a retrospective case series study. From January 2010 to December 2020, 79 severe burn patients complicated with AKI who received CRRT and met the inclusion criteria were admitted to the First Affiliated Hospital of Army Medical University (the Third Military Medical University). The general data (the same below) of all patients were collected, including gender, age, body mass index, burn area, burn index, cause of injury, whether combined with inhalation injury, acute physiology and chronic health status evaluation Ⅱ (APACHE Ⅱ) score and sepsis-related organ failure assessment (SOFA) score on admission, admission time after burn, and time of AKI after admission. The total efficacy of CRRT, including overall effective rate, complete effective rate, partial effective rate, ineffective rate, and deterioration rate, creatinine, urea, cystatin C, and fluid overload rate before and after treatment, in-hospital mortality, predictive mortality based on Baux scoring model, the most common cause of death, and length of hospital stay were recorded. According to the effect of CRRT, the patients were divided into effective group (42 patients) and ineffective group (37 patients). The general information of patients, the time to initiate CRRT after the occurrence of AKI, the duration of CRRT, etiology of AKI, AKI stage before CRRT initiation, CRRT mode, anticoagulant type, and in-hospital mortality were compared between the two groups of patients. The independent influencing factors for CRRT in severe burn patients complicated with AKI were screened. According to the etiology of AKI, the patients were divided into prerenal group (22 patients) and renal group (57 patients). The general information of patients, the time to initiate CRRT after the occurrence of AKI, the duration of CRRT, and total efficacy of CRRT (except for the most common cause of death) were compared between the two groups of patients.Results:Among the 79 patients, 73 cases were male and 6 cases were female, with age of (46±14) years, body mass index of (24.0±2.9) kg/m 2, total burn area of (69±26)% total body surface area (TBSA), full-thickness burn area of (44±25)%TBSA, and burn index of 57 (36, 76). There were 36 cases of flame burns, 19 cases of electrical burns, 16 cases of hydrothermal burns, 6 cases of explosive burns, and 2 cases of chemical burns. Thirty-nine patients were complicated with inhalation injury. The APACHE Ⅱ score was 16 (12, 18) and the SOFA score was 11 (5, 13) on admission. The patients were admitted to the hospital on 0 (0, 2) d after burn, and AKI occurred on 0 (0, 6) d after admission. The overall effective rate of CRRT was 53.16% (42/79), the complete effective rate was 30.38% (24/79), the partial effective rate was 22.78% (18/79), the ineffective rate was 31.65% (25/79), and the deterioration rate was 15.19% (12/79). The creatinine and urea of patients after treatment were significantly lower than those before treatment (with Z values of -3.26 and -2.54, respectively, P<0.05); there were no statistically significant differences in the cystatin C and fluid overload rate of patients before and after treatment ( P>0.05). The in-hospital mortality of patients was 17.72% (14/79), and the predictive mortality based on Baux scoring model was 75.10% (18.94%, 91.84%). The most common cause of death was multiple organ failure, and the length of hospital stay was 39.43 (11.52, 110.58) d. There were statistically significant differences in the full-thickness burn area, the duration of CRRT, and etiology of AKI of patients between effective group and ineffective group (with Z values of -1.99 and -2.90, respectively, χ2=5.58, P<0.05). There were no statistically significant differences in the other indicators ( P>0.05). The etiology of AKI and full-thickness burn area were the independent influencing factors for CRRT in severe burn patients complicated with AKI (with odds ratios of 4.21 and 1.03, respectively, 95% confidence intervals of 1.20-14.80 and 1.00-1.05, respectively, P<0.05). There were statistically significant differences in the cause of injury, overall effective rate of CRRT, total burn area, burn index, admission time after burn, time of AKI after admission, the time to initiate CRRT after the occurrence of AKI, and predictive mortality based on Baux score model of patients between prerenal group and renal group (with χ2 values of 12.59 and 5.58, respectively, Z values of 2.46, 2.43, -2.43, -4.03, -3.01, and -2.31, respectively, P<0.05). Before treatment, urea and cystatin C of patients in renal group were significantly higher than those in prerenal group (with Z values of -2.98 and -2.77, respectively, P<0.05), and the liquid overload rate was significantly lower than that in prerenal group ( Z=-2.99, P<0.05); after treatment, the cystatin C of patients in renal group was significantly higher than that in prerenal group ( Z=-2.08, P<0.05); there were no statistically significant differences in the other indicators ( P>0.05). Conclusions:CRRT can significantly improve renal function, avoid fluid overload, and alleviate renal injury in severe burn patients complicated with AKI. Prerenal AKI is the main independent influencing factor leading to ineffective CRRT.

Objective To investigate the changes in motor symptoms and cognitive levels in α-synuclein A53T transgenic mice at different months of age and possible mechanisms.Methods The α-synuclein A53T transgenic mice and wild C57/BL6J mice,aged 6 months,were selected.The open field test and the rotarod test were used to observe the motor function and emotional response of mice;fear conditioning and the novel object recognition test were used to observe the cognitive function of mice;Western blotting was used to measure the expression level of M1 cholinergic receptor in the hippocampus of the mice in the two groups,and the transcriptome method was used to observe the difference in gene ex-pression in the hippocampus between the two groups.Results Behavioral experiments showed that compared with the wild-type mice,the A53T transgenic mice showed cognitive dysfunction at 12 months of age[novel object recognition test:(36.74±13.33)vs(46.90±6.58),t=-2.368,P=0.027],and this phenomenon occurred before motor symptoms.West-ern blotting showed that there was no significant difference in the level of M1 cholinergic receptor in the hippocampus be-tween the A53T transgenic mice and the wild-type mice(P=0.537).The transcriptome analysis identified 290 differen-tially expressed genes between the two groups,among which there were 20 genes with significant difference,and the sig-nificantly downregulated gene sets in the hippocampus of A53T mice were mainly involved in anti-cellular inflammatory re-sponse and antioxidant level.Conclusion Cognitive dysfunction in A53T transgenic mice occurs earlier than motor symp-toms,with a relatively late time node.Transcriptome tests suggest that cellular inflammatory response and oxidative stress injury might be involved in the early pathological process of cognitive impairment in A53T mice.

This study was aimed at understanding the trends in,and scope of,severe fever with thrombocytopenia syndrome(SFTS)in Nanjing,analyzing the spatial distribution pattern,detecting high incidence cluster areas and key popula-tions,and scientifically guiding prevention and control strategies and measures.We obtained data on SFTS cases from 2010 to 2023 in Nanjing from the China Disease Control and Prevention Information System,and described the time,popu-lation,and spatial distribution characteristics.We used joinpoint regression to calculate the annual percentage change(APC)in incidence,then used FleXScan spatial clustering scan analysis to explore spatial clustering areas at the street level.A total of 507 SFTS cases were reported from 2010 to 2023 in Nanjing.The APC was 31.8％(95％CI:22.5％-41.9％,P＜0.001),and the reported incidence in 2023 was 1.42/100 000(134 cases).The seasonal indices from May to August were 2.7,2.1,3.0,and 1.3,respectively,accounting for 76.1％of the total cases.The median age was 66(IQR:55,73)years,which gradually increased from 59 years in 2010-2011 to 68 in 2022-2023(P＜0.001);94.1％of cases were in individuals 45 years or older.Farmers,homemakers/unemployed individuals,and retirees accounted for 90.1％.The epidemic area increased from 11 streets in four districts in 2010-2011 to 58 streets in 11 dis-tricts in 2022-2023.Except for 2012-2013,global spatial autocorrelation analysis showed positive Moran's I values(0.224-0.526,P＜0.001),and FlexScan scan indicated that several streets in Lishui District and Jiangning District were the most likely clusters.Four streets in Pukou District were the secondary clusters from 2018 to 2023,and three streets in Luhe District in 2022-2023 were the secondary clusters(all P＜0.05).The reported incidence of SFTS in Nanjing showed a rapid upward trend,with spread of epidemic areas.The spatial distribution pattern was clustered.Strengthened training in diagnosis and treatment technology and detection ability of medical institutions,surveillance in high-incidence areas,tracing of case flow,and health education of tick and disease prevention knowledge are recommended.