Objective To analyze the PLCβ4 gene mRNA expression and its clinical significance in hepatocellular carcinoma (HCC) based on TCGA database. Methods Based on the data on 424 clinical samples (including 374 cases of HCC tissues and 50 cases of nontumor liver tissues) in the TCGA database, Kaplan–Meier method, Cox regression analysis, and immune infiltration analysis were performed to evaluate the relationship between PLCβ4 gene and the clinical characteristics and survival prognosis of HCC patients. Correlation analysis between PLCβ4 gene and 24 types of immune cells was applied to investigate the relationship between PLCβ4 gene and immune cell infiltration and mRNA expression level of TP53 gene, a high-frequency mutation gene in HCC. In addition, paraffin sections of highly, moderately, and poorly differentiated tumor tissues and normal liver tissues from HCC patients were collected. The histopathological observation was carried out via HE staining method, and the expression levels of PLCβ4 and Ki-67 proteins in each clinical sample were verified through the immunohistochemical method. Results The expression level of PLCβ4 gene in HCC was significantly higher than that in normal tissues (P<0.01), and all patients in the PLCβ4 high-expression group had a significantly longer overall survival than those in the low-expression group (P<0.05), which suggested that PLCβ4 substantially affected the prognosis of HCC patients. Correlation analysis showed that the expression level of PLCβ4 gene was highly correlated with immune cell infiltration and the expression level of TP53 gene. As verified by clinical sample experiments, HE staining experiments and immunohistochemical results revealed that PLCβ4 gene expression in HCC tissue samples was significantly higher than that in normal tissues (P<0.001), and it was negatively correlated with the degree of differentiation. Conclusion PLCβ4 may serve as an independent prognostic factor in HCC and is expected to be a novel molecular target for HCC treatment.

Aim To investigate the intervention effect of Rehmannia radix water extract on bleomycin(BLM)-induced pulmonary fibrosis in mice combined with metabolomics and to reveal the potential mechanism,in order to provide new ideas for clinical treatment of pul-monary fibrosis.Methods Male C57BL/6N mice were randomly divided into the control group,model group,pirfenidone group(positive control,PFD,270 mg·kg-1),and low dose(DH-L,4.55 g·kg-1)group,medium dose(DH-M,9.1 g·kg-1)group and high dose(DH-H,18.2 g·kg-1)group of Rehman-nia.Except for the control group,BLM(5 mg·kg-1)was instilled into the trachea to establish the model of pulmonary fibrosis in the other groups.The survival rate,lung index and blood oxygen saturation of mice in each group were evaluated.HE and Masson staining were used to observe the pathological changes of lung tissue.WBP was used to detect lung function.Flow cytometry was used to detect the apoptosis of primary lung cells,ROS and immune cells.ELISA was used to detect the levels of fibrosis markers and inflammatory factors(α-SMA,collagen Ⅰ,collagen Ⅲ,TGF-β1,TNF-α,IL-1 β,and IL-6).Biochemical method was employed to detect the contents of GSH-Px,T-SOD and MDA.Liquid chromatograph mass spectrometer(LC-MS)metabolomics was used to analyze the changes of serum metabolic profile.Results Water extract of Re-hmannia significantly increased the survival rate,oxy-gen saturation and lung function of mice with pulmona-ry fibrosis,reduced the lung coefficient,ameliorated pathological damage and collagen deposition in lung tissue,reduced the levels of apoptosis and oxidative stress,and down-regulated the levels of inflammatory factors in lung tissue.It regulated the levels of metabo-lites such as bile acid metabolism,sphingolipid metabo-lism,and unsaturated fatty acid metabolism.Conclu-sions Water extract of Rehmannia inhibits lung injury and collagen deposition in mice with pulmonary fibrosis by inhibiting inflammatory response,which may be a-chieved by regulating the levels of inflammatory factors through the metabolic pathways of bile acid and sphin-golipid.

BACKGROUND:Ankle fracture complicated with deltoid ligament injury is clinically common,and one stage repair of deltoid ligament with internal fracture fixation has gradually become a main therapeutic method,which can significantly reduce the long-term complications of ankle joint.In recent years,new progress has been made in anatomical structure characteristics and dynamic biomechanical research of deltoid ligament,thus greatly improving repairing techniques for injury in the deep layer of deltoid ligament,but there are still some controversies.OBJECTIVE:To explore the clinical efficacy of suture anchor and medial malleolus repair in the treatment of ankle joint fractures with the deep layer of deltoid ligament.METHODS:A total of 56 patients with ankle joint fractures and complete fracture of the deep and superficial layer of deltoid ligament treated in Affiliated Haikou Hospital,Xiangya School of Medicine,Central South University from January 2017 to January 2022 were selected,and they were divided into two groups according to different treatment methods in repairing the deep layer of deltoid ligament with suture anchor:suture anchor repair group(n=32)and medial malleolus repair group(n=24).The medial clear space of ankle joint and American Orthopedic Foot and Ankle Society Score of patients in the two groups were evaluated before and after operative treatment.RESULTS AND CONCLUSION:(1)All the 56 patients finished the surgery smoothly and were followed up for more than 12 months after operation.Their ankle fracture healed,and the time for fracture healing was 8-12 weeks,with a mean of 10.5 weeks.(2)The medial clear space of ankle joint in the two groups 12 months after operation was remarkably narrower than that before operation,and the difference was statistically significant(P＜0.001).The medial clear space of ankle joint in the two groups maintained a normal range 12 months after operation,and there was no statistically significant difference between the two groups(P＞0.05).(3)The AOFAS scale of patients in the two groups 6 and 12 months after operation was obviously bigger than that before operation(P＜0.001),but there was no statistically significant difference in the American Orthopedic Foot and Ankle Society Score of patients between the two groups at corresponding time points(P＞0.05).(4)It is concluded that both suture anchor and medial malleolus repair in the treatment of injury in the deep layer of deltoid ligament can recover the medial clear space of ankle joint,effectively keep the stability of ankle,and thus achieve good clinical efficacy.

Objective:To investigate the impact of postoperative complications on adverse outcomes following curative-intent resection for gallbladder cancer (GBC).Methods:The multi-center real-world study was conducted. The clinicopathological data of 629 patients with GBC, who were admitted to 14 medical centers including The First Affiliated Hospital of Army Medical University from the national multicenter database of Biliary Surgery Group of Elite Group of Chinese Journal of Digestive Surgery, from April 2020 to April 2024 were collected. There were 225 males and 404 females, aged (64±10)years. Patients underwent open curative-intent resection for GBC. Observation indicators: (1)surgery, postoperative complica-tions and adverse outcomes; (2) analysis of risk factors affecting postoperative adverse outcomes in patients and population attributable fraction (PAF). Missing data in predictor variables were addressed using multiple imputation with chained equations, while cases with missing outcome variables were addressed using the "multiple imputation then deletion (MID)" strategy. The severity of multicollinearity among independent variables was assessed using the variance inflation factor (VIF) test. Multivariable possion regression models with log link and robust error variance were construc-ted incorporating restricted cubic splines (3 knots) to address nonlinear relationships in continuous variables, calculating adjusted relative risk ( RR) with corresponding 95% confidence interval ( CI). Adjusted PAF was calculated for each imputed dataset using the AF package of R software, with subsequent pooling performed according to Rubin's rules. Results:(1) Surgery, postoperative complications and adverse outcomes. All 629 patients underwent curative-intent resection for GBC, of which 143 cases had postoperative complications, including 68 cases of intra-abdominal ascites, 39 cases of pulmonary infection, 21 cases of bile leakage, 12 cases of intra-abdominal hemorrhage, 11 cases of liver failure, 10 cases of pan-creatic fistula, 10 cases of wound infection, 10 cases of gastroparesis, 7 cases of cholangitis, 7 cases of sepsis. The same patient could have more than one kind of complication. Of 629 patients, there were 19 cases of postoperative 90-day death and 11 cases of missing data, 42 cases with post-operative 90-day reoperation and 7 cases with missing data, 44 cases with postoperative 90-day readmission and 3 cases with missing data, 155 cases with prolonged postoperative hospital stay and 3 cases with missing data. (2) Analysis of risk factors affecting the postoperative adverse outcomes in patients and PAF. Results of multivariate analysis showed that pulmonary infection and liver failure were independent risk factors for postoperative 90-day mortality ( RR=3.74, 12.15, 95% CI as 1.18-11.83, 1.98-74.48, P<0.05). Pulmonary infection demons-trated the highest PAF as 4.61% (95% CI as 3.94%-5.28%, P<0.05). Intra-abdominal ascites, pulmonary infection, bile leakage, and intra-abdominal hemorrhage were independent risk factors for post-operative 90-day reoperation ( RR=4.80, 3.62, 3.46, 4.99, 95% CI as 2.49-9.26, 1.42-9.21, 1.34-8.92, 1.55-16.06, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 8.65% (95% CI as 8.22%-9.08%, P<0.05). Intra-abdominal ascites, bile leakage, and liver failure were independent risk factors for postoperative 90-day readmission ( RR=6.20, 3.33, 14.33, 95% CI as 3.21-11.95, 1.33-8.35, 3.72-55.28, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 9.11% (95% CI as 8.85%-9.37%, P<0.05). Intra-abdominal ascites, pulmonary infection, bile leakage, liver failure, and wound infection were independent risk factors for prolonged postoperative hospital stay ( RR=2.29, 2.21, 2.26, 2.14, 3.35, 95% CI as 1.63-3.23, 1.41-3.46, 1.32-3.86, 1.11-4.13, 1.70-6.60, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 6.03% (95% CI as 5.71%-6.35%, P<0.05). Conclusion:Pulmonary infection is the most significant risk factor for postoperative 90-day mortality after curative-intent resection for GBC, while intra-abdominal ascites is the most significant risk factor for postoperative 90-day reoperation, postoperative 90-day readmission, and prolonged postoperative hospital stay.

Objective To investigate 5-aminoimidazole-4-carboxamide ribonucleotide formyl transferase/inosine monophos-phate cyclohydrolase(ATIC),a key regulator of metabolism and cell proliferation,to explore its role in glioma proliferation,e-valuate its association with patient prognosis,and elucidate the underlying molecular mechanisms.Methods Using data from The Cancer Genome Atlas(TCGA),Genotype-Tissue Expression(GTEx),and Chinese Glioma Genome Atlas(CGGA)databas-es,we analyzed differential ATIC expression between tumor tissues and adjacent normal tissues in glioma patients,as well as its correlation with clinical features including pathological grade,isocitrate dehydrogenase(IDH)mutation status,and chromosome 1p/19q deletion.ATIC was knocked down using siRNA transfection.The effect of ATIC on the proliferation of glioma cell lines(LN229,U373,and U251)was evaluated using EdU,CCK-8,and colony formation assays.Furthermore,ATIC overexpression via plasmid transfection was analyzed in conjunction with flow cytometry and Western blotting analysis to assess cell cycle pro-gression and cyclin-related protein expression.Results ATIC expression was significantly elevated in glioma tissues compared to adjacent normal tissues(P＜0.01).Patients with high ATIC expression exhibited shorter overall survival(OS)and were asso-ciated with higher pathological grades,wild-type IDH status,and the presence of chromosome 1p/19q deletion.Compared with U373 and U251 glioma cell lines,LN229 and U87 glioma cell lines demonstrated higher ATIC expression.In siRNA-mediated ATIC knockdown models(siATIC-LN229,siATIC-U373),cell proliferation was suppressed as demonstrated by EdU,CCK-8,and colony formation assays,whereas ATIC overexpression in U251 cells promoted proliferation.Flow cytometry revealed G1-phase arrest and impaired S-phase progression in siATIC-LN229 cells.Conversely,ATIC overexpression in U251 cells decreased G1-phase accumulation and increased S-phase progression.Mechanistically,ATIC knockdown decreased the expression of phos-phorylated Rb(p-Rb),upregulated p21,and downregulated key cyclin-related proteins essential for G1/S transition.In contrast,ATIC overexpression facilitated the G1/S transition through p21 downregulation and enhanced phosphorylation of Rb pro-tein.Conclusion High ATIC expression is associated with poor clinical outcomes in glioma patients and may promote tumor progression through regulation of the p21-Rb signaling pathway.Therefore,ATIC represents a promising biomarker for both clinical diagnosis and prognosis in glioma.

Objective To determine KCNJ14 gene expression in human glioma cells and assess its impact on U251 cell pro-liferation,while exploring the potential mechanism involved.Methods The protein expression levels of KCNJ14 in different hu-man glioma cell lines(U87,U251,SNB19,and LN229)were analyzed by Western blotting.KCNJ14 was knocked down and over-expressed in U251 cells using siRNA and plasmid transfection,respectively.Subsequently,protein expression levels,cell prolif-eration capacity,and the regulation of cell cycle related proteins were measured in each group.The association between KCNJ14 mRNA expression and clinical survival prognosis in glioma patients was evaluated through statistical analysis of public databas-es.Results KCNJ14 protein expression varied across different human glioma cell lines,with the highest level observed in U251 cells.Inhibition of KCNJ14 suppressed U251 cell proliferation and impaired cell cycle progression.Bioinformatics analysis re-vealed that KCNJ14 mRNA expression was significantly associated with clinical characteristics and survival outcomes in glioma patients.Conclusion KCNJ14 exhibits differential expression in glioma cells and is negatively associated with patient prognosis.Mechanistically,it may regulate glioma cell proliferation by modulating cell cycle related proteins.

Creutzfeldt-Jakob disease(CJD)is a rapidly progressive and fatal neurodegenerative disorder caused by prions,with certain infectivity and iatrogenic transmission risks.With the rapid progress and application of new dia-gnostic biomarkers and detection methods,as well as the construction and improvement of surveillance and reporting systems,the detection of CJD in patients domestically and internationally has shown an increasing trend year by year.Due to its long incubation period and heterogeneity of early symptoms,early identification and diagnosis of the disease is difficult,increasing the risk of transmission within medical institutions.Currently,there is a lack of con-sensus on the infection prevention and control of CJD.In order to timely identify and diagnose CJD as well as effec-tively block its transmission in medical institutions,this consensus summarizes 15 clinical concerns and formulates 24 specific recommendations based on the latest domestic and international research findings and clinical evidence,as well as combines with clinical practice,aiming to standardize healthcare-associated infection prevention and control measures for CJD and reduce its transmission risk in medical institutions.

Objective To investigate 5-aminoimidazole-4-carboxamide ribonucleotide formyl transferase/inosine monophos-phate cyclohydrolase(ATIC),a key regulator of metabolism and cell proliferation,to explore its role in glioma proliferation,e-valuate its association with patient prognosis,and elucidate the underlying molecular mechanisms.Methods Using data from The Cancer Genome Atlas(TCGA),Genotype-Tissue Expression(GTEx),and Chinese Glioma Genome Atlas(CGGA)databas-es,we analyzed differential ATIC expression between tumor tissues and adjacent normal tissues in glioma patients,as well as its correlation with clinical features including pathological grade,isocitrate dehydrogenase(IDH)mutation status,and chromosome 1p/19q deletion.ATIC was knocked down using siRNA transfection.The effect of ATIC on the proliferation of glioma cell lines(LN229,U373,and U251)was evaluated using EdU,CCK-8,and colony formation assays.Furthermore,ATIC overexpression via plasmid transfection was analyzed in conjunction with flow cytometry and Western blotting analysis to assess cell cycle pro-gression and cyclin-related protein expression.Results ATIC expression was significantly elevated in glioma tissues compared to adjacent normal tissues(P＜0.01).Patients with high ATIC expression exhibited shorter overall survival(OS)and were asso-ciated with higher pathological grades,wild-type IDH status,and the presence of chromosome 1p/19q deletion.Compared with U373 and U251 glioma cell lines,LN229 and U87 glioma cell lines demonstrated higher ATIC expression.In siRNA-mediated ATIC knockdown models(siATIC-LN229,siATIC-U373),cell proliferation was suppressed as demonstrated by EdU,CCK-8,and colony formation assays,whereas ATIC overexpression in U251 cells promoted proliferation.Flow cytometry revealed G1-phase arrest and impaired S-phase progression in siATIC-LN229 cells.Conversely,ATIC overexpression in U251 cells decreased G1-phase accumulation and increased S-phase progression.Mechanistically,ATIC knockdown decreased the expression of phos-phorylated Rb(p-Rb),upregulated p21,and downregulated key cyclin-related proteins essential for G1/S transition.In contrast,ATIC overexpression facilitated the G1/S transition through p21 downregulation and enhanced phosphorylation of Rb pro-tein.Conclusion High ATIC expression is associated with poor clinical outcomes in glioma patients and may promote tumor progression through regulation of the p21-Rb signaling pathway.Therefore,ATIC represents a promising biomarker for both clinical diagnosis and prognosis in glioma.

Objective To determine KCNJ14 gene expression in human glioma cells and assess its impact on U251 cell pro-liferation,while exploring the potential mechanism involved.Methods The protein expression levels of KCNJ14 in different hu-man glioma cell lines(U87,U251,SNB19,and LN229)were analyzed by Western blotting.KCNJ14 was knocked down and over-expressed in U251 cells using siRNA and plasmid transfection,respectively.Subsequently,protein expression levels,cell prolif-eration capacity,and the regulation of cell cycle related proteins were measured in each group.The association between KCNJ14 mRNA expression and clinical survival prognosis in glioma patients was evaluated through statistical analysis of public databas-es.Results KCNJ14 protein expression varied across different human glioma cell lines,with the highest level observed in U251 cells.Inhibition of KCNJ14 suppressed U251 cell proliferation and impaired cell cycle progression.Bioinformatics analysis re-vealed that KCNJ14 mRNA expression was significantly associated with clinical characteristics and survival outcomes in glioma patients.Conclusion KCNJ14 exhibits differential expression in glioma cells and is negatively associated with patient prognosis.Mechanistically,it may regulate glioma cell proliferation by modulating cell cycle related proteins.

Creutzfeldt-Jakob disease(CJD)is a rapidly progressive and fatal neurodegenerative disorder caused by prions,with certain infectivity and iatrogenic transmission risks.With the rapid progress and application of new dia-gnostic biomarkers and detection methods,as well as the construction and improvement of surveillance and reporting systems,the detection of CJD in patients domestically and internationally has shown an increasing trend year by year.Due to its long incubation period and heterogeneity of early symptoms,early identification and diagnosis of the disease is difficult,increasing the risk of transmission within medical institutions.Currently,there is a lack of con-sensus on the infection prevention and control of CJD.In order to timely identify and diagnose CJD as well as effec-tively block its transmission in medical institutions,this consensus summarizes 15 clinical concerns and formulates 24 specific recommendations based on the latest domestic and international research findings and clinical evidence,as well as combines with clinical practice,aiming to standardize healthcare-associated infection prevention and control measures for CJD and reduce its transmission risk in medical institutions.