Metabolic dysfunction-associated steatohepatitis (MASH), a severe type of metabolic dysfunction-associated steatotic liver disease (MASLD), is a leading etiology of end-stage liver disease worldwide, posing significant health and economic burdens. microRNA-320 (miR-320), a ubiquitously expressed and evolutionarily conserved miRNA, has been reported to regulate lipid metabolism; however, whether and how miR-320 affects MASH development remains unclear. By performing miR-320 in situ hybridization with RNAscope, we observed a notable downregulation of miR-320 in hepatocytes during MASH, correlating with disease severity. Most importantly, miR-320 downregulation in hepatocytes exacerbated MASH progression as demonstrated that hepatocyte-specific miR-320 deficient mice were more susceptible to high-fat, high-fructose, high-cholesterol diet (HFHC) or choline-deficient, amino acid-defined, high-fat diet (CDAHFD)-induced MASH compared with control littermates. Conversely, restoration of miR-320 in hepatocytes ameliorated MASH-related steatosis and fibrosis by injection of adeno-associated virus 8 (AAV8) carrying miR-320 in different types of diet-induced MASH models. Mechanistic studies revealed that miR-320 specifically regulated fibroblast growth factor 1 (FGF1) production in hepatocytes by inhibiting regulator factor X1 (RFX1) expression. Notably, knockdown of Rfx1 in hepatocytes mitigated MASH by enhancing FGF1-mediated AMPK activation. Our findings underscore the therapeutic potential of hepatic miR-320 supplementation in MASH treatment by inhibiting RFX1-mediated FGF1 suppression.

In recent decades, the prevalence of hyperuricemia and gout has increased dramatically due to lifestyle changes. The drugs currently recommended for hyperuricemia are associated with adverse reactions that limit their clinical use. In this study, we report that berberine (BBR) is an effective drug candidate for the treatment of hyperuricemia, with its mechanism potentially involving the modulation of gut microbiota and its metabolite, succinic acid. BBR has demonstrated good therapeutic effects in both acute and chronic animal models of hyperuricemia. In a clinical trial, oral administration of BBR for 6 months reduced blood uric acid levels in 22 participants by modulating the gut microbiota, which led to an increase in the abundance of Bacteroides and a decrease in Clostridium sensu stricto_1. Furthermore, Bacteroides fragilis was transplanted into ICR mice, and the results showed that Bacteroides fragilis exerted a therapeutic effect on uric acid similar to that of BBR. Notably, succinic acid, a metabolite of Bacteroides, significantly reduced uric acid levels. Subsequent cell and animal experiments revealed that the intestinal metabolite, succinic acid, regulated the upstream uric acid synthesis pathway in the liver by inhibiting adenosine monophosphate deaminase 2 (AMPD2), an enzyme responsible for converting adenosine monophosphate (AMP) to inosine monophosphate (IMP). This inhibition resulted in a decrease in IMP levels and an increase in phosphate levels. The reduction in IMP led to a decreased downstream production of hypoxanthine, xanthine, and uric acid. BBR also demonstrated excellent renoprotective effects, improving nephropathy associated with hyperuricemia. In summary, BBR has the potential to be an effective treatment for hyperuricemia through the gut-liver axis.

Purpose To investigate the expression of ubiquitin-specific protease 9X(USP9X)and programmed cell death ligand 1(PD-L1)in pulmonary sarcomatoid carcinoma(PSC),and to assess their clinicopathological signif-icance.Methods Immunohistochemical staining was performed on 48 PSC specimens using the EnVision two-step method to detect USP9X and PD-L1 expression.The relationships between their expressions levels and clinicopathologi-cal parameters were analyzed.Results USP9X was positively expressed in 81.3％(39/48)of PSC cases,and the USP9X positivity rate was significantly higher in lymph node-negative tumors compared with lymph node-positive tumors(P＜0.05).PD-L1 expression was observed in 70.8％(34/48)of cases,with a significantly higher positivity rate in male patients than in female patients(P＜0.01).Correlation analysis showed a significant positive association between USP9X and PD-L1 expression in PSC(P＜0.05).Conclusion Both USP9X and PD-L1 are highly expressed in PSC,and their expression levels are significantly positively correlated.Combined detection of USP9X and PD-L1 may aid in predicting the efficacy of immunotherapy in PSC.

In view of the characteristics of H5N6 subtype avian influenza virus(AIV)that it has both high pathogenicity and the risk of cross-species transmission,posing a serious threat to the poultry farming industry and public health security,in order to effectively prevent and control the spread of H5N6 avian influenza,a rapid,sensitive and specific detection technolo-gy was established in this study.The specific monoclonal antibodies against the neuraminidase N6 protein of avian influenza A virus subtype H5N6 were obtained through hybridoma and monoclonal antibody technology.These antibodies were coupled and labeled with carboxyl-functionalized fluorescent quantum dots,along with previously prepared specific antibodies against the hemagglutinin H5 protein.A rapid fluorescence immunochromatographic detection method for the H5N6 subtype of avian influ-enza virus was established according to the principle of double-antibody sandwich immunochromatography.This method a-chieved a detection sensitivity of 1 ng/mL for recombinant hemagglutinin H5 subtype protein and 0.1 ng/mL for recombinant neuraminidase N6 subtype protein.Moreover,the method exhibited no cross-reactivity with other influenza subtypes or patho-gens,such as Newcastle disease(ND),infectious bronchitis(IB),and infectious laryngotracheitis(ILT),thus demonstrating good specificity.The method effectively identified the highly pathogenic avian influenza virus H5 subtype and directly distin-guished the H5N6 subtype with good accuracy.The fluorescent quantum dot immunochromatographic typing detection method established herein met the sensitivity,specificity,and accuracy requirements for H5N6 subtype detection,and can be further used for rapid detection of the H5 and H5N6 subtypes of avian influenza virus.

Objective:To evaluate preoperative high-resolution thin-layer cervical enhanced CT used to predict the venous route of the submental flap reflux vein and its relationship with adjacent structures in order to guide the anatomical understanding and protection of submental flap in pharyngeal cancer surgery.Methods:Sixty consecutive patients with pharyngeal cancer who underwent submental flap repair surgery in our department from March 2022 to December 2024, as well as 60 patients who were accepted neck dissection suffering other cancers, were selected. Before surgery, high-resolution cervical enhanced CT scans were performed, and the position of the transverse section of the facial vein in the venous phase horizontal image gradually variation tendency was focused layer by layer. The direction and adjacent relationship of the submental flap reflux veins were determined and recorded. Combined with 60 patients with other head and neck tumors who underwent neck dissection in our department during the same period (a total of 120 cases, 240 sides), the classification and management of the draining veins of Fang′s mental flap were conducted. Type Ⅰ mainly drains into the internal jugular vein; Type Ⅱ mainly drains into the external jugular vein and Type Ⅲ mainly drains into the anterior jugular vein (often accompanied by an external jugular draining branch). The status and proportion of venous drainage were analyzed.Results:Vascular predictive coincidence rate was 98.3% (59/60) among the 60 patients with pharyngeal cancer. Only one patient was predicted to have a simple return to the external jugular vein. However, during the operation, in addition to the main return to the external jugular vein, a small portion also returned to the internal jugular vein. Submental flap reflux vessels were classified into three types based on intraoperative submental flap venous return in 60 cases of laryngopharyngeal cancer, in conjunction with the analysis of venous return patterns from 240 cervical CT scans. Type Ⅰ mainly refluxed to the internal jugular vein, accounting for 42.1%. Type Ⅱ mainly refluxed to the external jugular vein (47.9%). Type Ⅲ mainly refluxed to the anterior jugular vein (10.0%). The total detection rate of CT reading of 240 venous reflux was 98.7% (237/240). Vascular predictive coincidence rate was 97.9%(235/240).Conclusion:The detailed analysis of submental venous return vessels can accurately predict the direction of reflux veins and its surrounding areas by preoperative high-resolution enhanced CT scan. This provides reliable guidance for the anatomy and protection of the submental flap reflux veins during surgery.

Objective:To compare the survival outcomes of different subsequent treatment regimens in patients with locally advanced hypopharyngeal squamous cell carcinoma (HPSCC) who achieved partial response (PR) after neoadjuvant chemotherapy based on the gross tumor volume regression rate (GTVRR).Methods:This retrospective study included patients with locally advanced HPSCC treated at the Department of Head and Neck Surgery, Beijing Tongren Hospital, from January 2011 to December 2023. The cohort included 135 males and 3 females, aged from 35 to 77 years. All patients received 2-3 cycles of TPF regimen (paclitaxel+cisplatin+5-fluorouracil) neoadjuvant chemotherapy. Subsequent treatments included concurrent chemoradiotherapy or surgery combined with postoperative adjuvant radiotherapy. The impacts of different subsequent treatment modalities on the survivals and prognoses of patients were compared based on GTVRR thresholds of 50% and 70%. The χ 2 test was used to analyze influencing factors; survival analysis and intergroup comparisons were performed using the Kaplan-Meier method and Log-rank test; prognostic factors were assessed using univariate and multivariate Cox regression analyses. Results:The 5-year OS and PFS rates were 56.5% and 47.9%, respectively, while, the 10-year OS and PFS rates were 25.8% and 21.2%, respectively. The median OS was 75 months, and the median PFS was 48 months. The laryngeal function preservation rate for the entire cohort was 83.3%. The patients who underwent surgery combined with postoperative radiotherapy had significantly better OS and PFS outcomes than those treated with concurrent chemoradiotherapy ( P<0.05). Stratification based on GTVRR revealed that the surgery plus postoperative radiotherapy regimen was particularly effective for PR patients with a GTVRR of 30%-70%, showing significantly better OS and PFS compared to the concurrent chemoradiotherapy group ( P<0.05). Conclusion:The optimal subsequent treatment for PR-HPSCC may be surgery-based comprehensive treatment, particularly for patients with a GTVRR of 30%-70%. This study offers valuable insights for the stratified treatment of HPSCC, which could contribute to improving overall patient prognosis.

Objective To explore the relationship between the expression of cytochrome P450 family 2 subfamily W member 1(CYP2W1)and microRNA-491-5p(miR-491-5p)in hepatocellular cancer(HCC)tissues and clinical pathological characteristics and prognosis.Methods One hunderd and twenty HCC patients who underwent surgical treatment in our hospital from February 2019 to February 2021 were selected as the observation subjects,and the HCC tissues and adjacent tissues specimens were collected during the surgical process.The quantitative real-time PCR(qRT-PCR)was used to detect mRNA expression levels and the immunohistochemistry was used to detect protein expression levels.The differences in miR-491-5p and CYP2W1 mRNA expression between HCC tissues and adjacent tissues,as well as the correlation between their expression and clinical pathological characteristics of patients,were analyzed.The Kaplan-Meier survival analysis curves were performed.Results The expression level of CYP2W1 mRNA in HCC tissues was significantly higher than that in adjacent tissues,and the expression level of miR-491-5p was significantly lower than that in adjacent tissues(P<0.05).The positive expression rate of CYP2W1 protein in HCC tissues was significantly higher than that in adjacent tissues(χ2=23.721,P<0.001).The expression levels of miR-491-5p and CYP2W1 mRNA in HCC tissues were negatively correlated(r=-0.538,P<0.001).The expression of CYP2W1 and miR-491-5p was associated with TNM staging,tumor differentiation,lymph node metastasis,and pathological grading in HCC(P<0.05).The Kaplan-Meier survival curves analysis revealed that the 3-year cumulative survival rate in the CYP2W1 positive-expression group was 31.4%,which was lower than that in the CYP2W1 negative-expression group(63.8%)(log-rank χ2=12.243,P=0.000).The 3-year cumulative survival rate in the miR-491-5p high-expression group was 61.3%,which was significantly higher than that in the miR-491-5p low-expression group(37.9%)(log-rank χ2=4.490,P=0.034).The expression level of CYP2W1 mRNA in the recurrent group was higher than that in the non-recurrent group,and the expression level of miR-491-5p was lower than that in the non-recurrent group(P<0.05).The expression level of CYP2W1 mRNA in the metastatic group was higher than that in the non-metastatic group,and the expression level of miR-491-5p was lower than that in the non-metastatic group(P<0.05).Conclusion CYP2W1 is highly expressed while miR-491-5p is lowly expressed in HCC tissues,and both are significantly associated with certain clinicopathological features and prognosis of HCC patients.

Objective To explore the relationship between the expression of cytochrome P450 family 2 subfamily W member 1(CYP2W1)and microRNA-491-5p(miR-491-5p)in hepatocellular cancer(HCC)tissues and clinical pathological characteristics and prognosis.Methods One hunderd and twenty HCC patients who underwent surgical treatment in our hospital from February 2019 to February 2021 were selected as the observation subjects,and the HCC tissues and adjacent tissues specimens were collected during the surgical process.The quantitative real-time PCR(qRT-PCR)was used to detect mRNA expression levels and the immunohistochemistry was used to detect protein expression levels.The differences in miR-491-5p and CYP2W1 mRNA expression between HCC tissues and adjacent tissues,as well as the correlation between their expression and clinical pathological characteristics of patients,were analyzed.The Kaplan-Meier survival analysis curves were performed.Results The expression level of CYP2W1 mRNA in HCC tissues was significantly higher than that in adjacent tissues,and the expression level of miR-491-5p was significantly lower than that in adjacent tissues(P<0.05).The positive expression rate of CYP2W1 protein in HCC tissues was significantly higher than that in adjacent tissues(χ2=23.721,P<0.001).The expression levels of miR-491-5p and CYP2W1 mRNA in HCC tissues were negatively correlated(r=-0.538,P<0.001).The expression of CYP2W1 and miR-491-5p was associated with TNM staging,tumor differentiation,lymph node metastasis,and pathological grading in HCC(P<0.05).The Kaplan-Meier survival curves analysis revealed that the 3-year cumulative survival rate in the CYP2W1 positive-expression group was 31.4%,which was lower than that in the CYP2W1 negative-expression group(63.8%)(log-rank χ2=12.243,P=0.000).The 3-year cumulative survival rate in the miR-491-5p high-expression group was 61.3%,which was significantly higher than that in the miR-491-5p low-expression group(37.9%)(log-rank χ2=4.490,P=0.034).The expression level of CYP2W1 mRNA in the recurrent group was higher than that in the non-recurrent group,and the expression level of miR-491-5p was lower than that in the non-recurrent group(P<0.05).The expression level of CYP2W1 mRNA in the metastatic group was higher than that in the non-metastatic group,and the expression level of miR-491-5p was lower than that in the non-metastatic group(P<0.05).Conclusion CYP2W1 is highly expressed while miR-491-5p is lowly expressed in HCC tissues,and both are significantly associated with certain clinicopathological features and prognosis of HCC patients.

In view of the characteristics of H5N6 subtype avian influenza virus(AIV)that it has both high pathogenicity and the risk of cross-species transmission,posing a serious threat to the poultry farming industry and public health security,in order to effectively prevent and control the spread of H5N6 avian influenza,a rapid,sensitive and specific detection technolo-gy was established in this study.The specific monoclonal antibodies against the neuraminidase N6 protein of avian influenza A virus subtype H5N6 were obtained through hybridoma and monoclonal antibody technology.These antibodies were coupled and labeled with carboxyl-functionalized fluorescent quantum dots,along with previously prepared specific antibodies against the hemagglutinin H5 protein.A rapid fluorescence immunochromatographic detection method for the H5N6 subtype of avian influ-enza virus was established according to the principle of double-antibody sandwich immunochromatography.This method a-chieved a detection sensitivity of 1 ng/mL for recombinant hemagglutinin H5 subtype protein and 0.1 ng/mL for recombinant neuraminidase N6 subtype protein.Moreover,the method exhibited no cross-reactivity with other influenza subtypes or patho-gens,such as Newcastle disease(ND),infectious bronchitis(IB),and infectious laryngotracheitis(ILT),thus demonstrating good specificity.The method effectively identified the highly pathogenic avian influenza virus H5 subtype and directly distin-guished the H5N6 subtype with good accuracy.The fluorescent quantum dot immunochromatographic typing detection method established herein met the sensitivity,specificity,and accuracy requirements for H5N6 subtype detection,and can be further used for rapid detection of the H5 and H5N6 subtypes of avian influenza virus.

Purpose To investigate the expression of ubiquitin-specific protease 9X(USP9X)and programmed cell death ligand 1(PD-L1)in pulmonary sarcomatoid carcinoma(PSC),and to assess their clinicopathological signif-icance.Methods Immunohistochemical staining was performed on 48 PSC specimens using the EnVision two-step method to detect USP9X and PD-L1 expression.The relationships between their expressions levels and clinicopathologi-cal parameters were analyzed.Results USP9X was positively expressed in 81.3％(39/48)of PSC cases,and the USP9X positivity rate was significantly higher in lymph node-negative tumors compared with lymph node-positive tumors(P＜0.05).PD-L1 expression was observed in 70.8％(34/48)of cases,with a significantly higher positivity rate in male patients than in female patients(P＜0.01).Correlation analysis showed a significant positive association between USP9X and PD-L1 expression in PSC(P＜0.05).Conclusion Both USP9X and PD-L1 are highly expressed in PSC,and their expression levels are significantly positively correlated.Combined detection of USP9X and PD-L1 may aid in predicting the efficacy of immunotherapy in PSC.