ObjectiveTo observe the effect of M1 bone marrow-derived macrophages （M1-BMDM） with Wnt5a knockdown on liver fibrosis and regeneration in a rat model of liver cirrhosis， and to investigate its gain-of-function effect compared with unmodified M1-BMDM. MethodsPrimary bone marrow-derived macrophages were isolated from rats and were polarized to M1 phenotype to construct M1-BMDM^Wnt5a-KD cells. A rat model of liver cirrhosis induced by CCl₄/2-AAF was established， and at the end of week 8， rats were randomly divided into model group， M1-BMDM group， M1-BMDM Wnt5a-knockdown empty vector group （M1-BMDM^KD-EV group）， and M1-BMDM Wnt5a-knockdown group （M1-BMDM^Wnt5a-KD group）， with 6 rats in each group. On the first day of week 9， the rats in each group were given a single injection of the corresponding cells via the caudal vein， along with an intraperitoneal injection of a CCR2 inhibitor. Six rats without any treatment were used as normal control group. Samples were collected at the end of week 12 to assess liver histopathology， serum liver function parameters， hepatic stellate cell activation， and the expression levels of mature hepatocyte markers. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the model group， all cell treatment groups had significant alleviation of liver inflammatory response and significant reductions in the activities of alanine aminotransferase and aspartate aminotransferase （AST） in serum （all P<0.01）， and the M1-BMDM^Wnt5a-KD group had a significantly lower serum level of AST than the M1-BMDM group （P<0.05）. The semi-quantitative analysis based on immunohistochemical staining showed that compared with the model group， all cell treatment groups had a significant reduction in the percentage of CD68-positive area （all P<0.05）， and compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had a significant reduction in the percentage of CD68-positive area and a significant increase in the percentage of CD163-positive area （both P<0.05）. Compared with the model group， all cell treatment groups had significant reductions in the mRNA expression levels of CD68 and tumor necrosis factor-α （all P<0.05） and the protein expression level of CD68 （all P<0.01）； compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significant increases in the protein and mRNA expression levels of CD163 （both P<0.05）， significant reductions in the protein and mRNA expression levels of CD68 （both P<0.05）， and a significant reduction in the protein expression level of tumor necrosis factor-α （P<0.01）. Sirius Red collagen staining and alpha-smooth muscle actin （α-SMA） immunohistochemical staining showed that compared with the model group， all cell treatment groups had significant alleviation of liver collagen deposition and α-SMA-positive area， with the most significant changes in the M1-BMDM^Wnt5a-KD group， and compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significantly smaller Sirius Red-positive area and α-SMA-positive area and a significantly lower content of hydroxyproline in liver tissue （all P<0.05）. Compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significant reductions in the protein and mRNA expression levels of α-SMA and the mRNA expression level of COL-I and TGF-β （all P<0.05）. Compared with the model group， all cell treatment groups had a significant increase in the protein expression level of HNF-4α in liver tissue （all P<0.05）， and the M1-BMDM^Wnt5a-KD group had significantly higher protein and mRNA expression levels of HNF-4α and hepatocyte specific antigen than the M1-BMDM^KD-EV group （both P<0.05）. The M1-BMDM^Wnt5a-KD group had a significantly higher serum level of albumin than the M1-BMDM^KD-EV group （P<0.01）. Immunofluorescence co-staining showed that compared with the model group， all cell treatment groups had a significant increase in the number of cells stained positive for HNF and HNF-4α and Ki67 （all P<0.01）， and the M1-BMDM^Wnt5a-KD group had a significantly higher number of such cells than the M1-BMDM^KD-EV group （P<0.05）. ConclusionInhibition of Wnt5a expression enhances the therapeutic effect of M1-BMDM on rats with liver cirrhosis induced by CCl₄/2-AAF， which provides new ideas for enhancing the anti-cirrhotic effect of M1-BMDM through genetic modification.

Objective:To investigate the prognosis of postoperative adjuvant therapy for hepatocellular carcinoma after conversion therapy of combined targeted therapy and immunotherapy followed by sequential hepatectomy.Methods:The retrospective cohort study was conducted. The clinicopathological data of 103 patients with initially unresectable hepatocellular carcinoma (HCC) who were admitted to 11 medical centers in China, including Mengchao Hepatobiliary Hospital of Fujian Medical University et al, from November 2019 to May 2023 were collected. There were 83 males and 20 females, aged (54±12)years. All 103 patients underwent conversion therapy of tyrosine kinase inhibitors (TKIs) and immune checkpoint inhibitors (ICIs) successfully followed by sequential hepatectomy, of which 72 patients undergoing postoperative adjuvant therapy were divided into the adjuvant therapy group, and 31 patients undergoing postoperative follow-up monitoring were divided into the follow-up monitoring group. Observation indicators: (1) follow-up and postoperative condi-tions; (2) analysis of factors influencing recurrence-free survival time of patients; (3) stratified ana-lysis. Comparison of count data between group was conducted using the chi-square test or Fisher exact probability. The R software was used to draw survival curves, and the Log-rank test was used for survival analysis. Univariate and multivariate analyses were conducted using the Cox proportional hazard model. Results:(1) Follow-up and postoperative conditions. All 103 patients were followed up for 21.0(range, 1.9?47.2)months, with the median recurrence-free survival time of 28.7 months and the 1-, 2-, 3-year recurrence-free survival rates of 68.6%, 55.6%, 41.2%. The median overall survival time of 103 patients was unreached, and the 1-, 2-, 3-year overall survival rates were 90.9%, 82.1%, 69.6%, respectively. The median recurrence-free survival time was 33.1 months in patients of the adjuvant therapy group, with the 1-, 2-year recurrence-free survival rates as 77.2%, 61.5%. The median recurrence-free survival time was 11.1 months in patients of the follow-up monitoring group, with the 1-, 2-year recurrence-free survival rates as 46.6%, 40.8%. There was a significant difference in recurrence-free survival between the two groups of patients ( χ2=5.492, P<0.05). (2) Analysis of factors influencing recurrence-free survival time of patients. Results of multivariate analy-sis showed that pathologic complete response and postoperative adjuvant therapy were independent factors influencing recurrence-free survival time of HCC patients undergoing conversion therapy of combined targeted therapy and immunotherapy followed by sequential hepatectomy ( hazard ratio=0.297, 0.492, 95% confidence interval as 0.137?0.647, 0.268?0.903, P<0.05). (3) Stratified analysis. Of the 71 patients with non-pathologic complete response, the median recurrence-free survival time of 48 patients in the adjuvant therapy group was 24.0 months, with the 1-, 2-year recurrence-free survival rates as 67.4%, 48.8%. The median recurrence-free survival time of 23 patients with non-pathological complete response in the follow-up monitoring group was 7.4 months, with the 1-, 2-year recurrence-free survival rates as 35.0%, 26.3%. There was a significant difference in recurrence-free survival between the 48 patients with non-pathologic complete response in the adjuvant therapy group and the 23 patients with non-pathologic complete response in the follow-up monitoring group ( χ2=5.241, P<0.05). Conclusion:For HCC patients with conversion therapy of TKIs and ICIs followed by sequential hepatectomy, postoperative adjuvant therapy, compared to postoperative follow-up monitoring, can prolong the recurrence-free survival time of patients, of whom cases with non-pathologic complete response can benefit from adjuvant therapy.

Microfibrino-associated protein (MFAP) 2 is an extracellular matrix glycoprotein and a member of the MFAP family. It participates in the assembly of extracellular elastic microfibers.Upregulation of MFAP2 can promote the occurrence and development of various tumors and regulate multiple cancer-related signaling pathways and is related to their prognosis, making it a potential target for tumor treatment. This article summarizes the research progress on the pathogenesis, targeted therapy and prognosis of MFAP2 in malignant tumors.

Objective To investigate the association of serum exosomal microRNAs(miRNAs)with hepatic inflammatory injury and histological outcomes in patients with chronic hepatitis B(CHB).Methods Peripheral serum samples were collected from six healthy adults and six patients with CHB,and size exclusion chromatography was used to extract exosomes.Small RNA sequencing and transcriptomic analysis were used to identify the serum exosomal miRNAs associated with liver inflammatory injury and fibrosis,and quantitative real-time PCR was used for validation in a mouse model of acute liver injury induced by lipopolysaccharide/D-galactosamine,a rat model of liver fibrosis induced by carbon tetrachloride,and 84 CHB patients undergoing liver biopsy twice before and after treatment.The independent-samples t test was used for comparison of normally distributed continuous data between two groups;an analysis of variance was used for comparison between multiple groups,and the Tukey test was used for further comparison between two groups.The Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups;the Kruskal-Wallis H test was used for comparison between multiple groups,and the Dunn test was used for further comparison between two groups.The chi-square test or the Fisher's exact test was used for comparison of categorical data between groups.The univariate and multivariate Logistic regression analyses were used to investigate influencing factors.Results Abnormal expression of serum exosomal miR-122-5p was observed in patients with CHB,and it was downregulated in patients with confluent necrosis and advanced fibrosis.In the mouse model of acute liver injury and the rat model of liver fibrosis,compared with the control group,the model group had a significant reduction in the expression level of miR-122-5p in the liver(P=0.048 and 0.014),and compared with the patients with mild liver injury,the patients with severe confluent necrosis and advanced fibrosis showed a significant reduction in the expression level of miR-122-5p in liver tissue(P<0.05).Among the 84 CHB patients,the patients with severe hepatic confluent necrosis or advanced liver fibrosis had a significantly lower expression level of serum exosomal miR-122-5p than those with mild liver injury(P<0.001 and P=0.003).The multivariate Logistic regression analysis showed that the expression level of miR-122-5p was an independent influencing factor for confluent necrosis(odds ratio[OR]=0.001,95%confidence interval[CI]:0.000-0.037,P=0.005)and liver fibrosis degree(OR=0.568,95%CI:0.331-0.856,P=0.019).In addition,compared with the patients with low expression of miR-122-5p,the patients with high expression of miR-122-5p before treatment had a significantly higher reversal rate of liver fibrosis after 72 weeks of antiviral therapy(64.3%vs 38.1%,P=0.029).Conclusion Serum exosomal miR-122-5p in CHB patients is closely associated with the progression of hepatic confluent necrosis and fibrosis,and the reduction in the expression level of miR-122-5p may aggravate hepatic confluent necrosis,promote the progression of fibrosis,and affect the histological outcome of CHB patients after antiviral therapy.

Microfibrino-associated protein (MFAP) 2 is an extracellular matrix glycoprotein and a member of the MFAP family. It participates in the assembly of extracellular elastic microfibers.Upregulation of MFAP2 can promote the occurrence and development of various tumors and regulate multiple cancer-related signaling pathways and is related to their prognosis, making it a potential target for tumor treatment. This article summarizes the research progress on the pathogenesis, targeted therapy and prognosis of MFAP2 in malignant tumors.

A 20-year-old male patient with T-lymphoblastic lymphoma/leukemia received 9/10 human leukocyte antigen-compatible unrelated peripheral blood stem cell transplantation. He was transplanted with 5.91×10 8 mononuclear cells/kg and 2.88×10 6 CD34 + cells/kg, and neutrophil engraftment was obtained at +11 days and platelet engraftment at +9 days. After transplantation, he presented with repeatedly increased serum creatinine levels, BK virus (BKV) -associated hemorrhagic cystitis, and BKV viremia. BK virus nephropathy was diagnosed based on renal biopsy and metagenomic next-generation sequencing. After adjusting the immunosuppressant, intravenous immunoglobulin, and donor lymphocyte infusion treatment, the patient’s renal function deteriorated progressively, and he eventually died of multiple organ failure at +289 days.

Objective To investigate the correlation between serum microRNA-210(miR-210),tumor necrosis factor-stimulated gene 6(TSG-6),and complement C1q tumor necrosis factor-related protein 3(CTRP3)and their association with myocardial fibrosis and prognosis in patients with dilated cardiomyopathy(DCM).Methods A total of 117 patients with DCM admitted to Taizhou People's Hospital between March and August 2024 were enrolled in the DCM group.Based on cardiac magnetic resonance imaging findings,these patients were further classified into a myocardial fibrosis group(n=96)and a non-fibrosis group(n=21).Additionally,according to the occurrence of acute heart failure during one-year follow-up,they were categorized into a heart failure group(n=47)and a non-heart failure group(n=70).Concurrently,58 age-and sex-matched healthy volunteers were recruited as the control group.Serum levels of miR-210,TSG-6,CTRP3,N-terminal propeptide of type Ⅲ procollagen(PⅢNP),left ventricular ejection fraction(LVEF),and N-terminal pro-B-type natriuretic peptide(NT-proBNP)were measured and compared across all groups.Results The DCM group exhibited significantly higher serum levels of miR-210,TSG-6,PⅢNP,and NT-proBNP,lower CTRP3 levels,and reduced LVEF compared to the healthy controls(P<0.05).Similarly,the fibrosis group showed elevated serum levels of miR-210,TSG-6,PⅢNP,and NT-proBNP,decreased CTRP3 levels,and impaired LVEF relative to the non-fibrosis group(P<0.05).The heart failure group also demonstrated higher serum concentrations of these biomarkers,along with lower CTRP3 and reduced LVEF,compared to the non-heart failure group(P<0.05).Serum miR-210 and TSG-6 levels were positively correlated with PⅢNP and NT-proBNP(P<0.05)and negatively correlated with LVEF(P<0.05).Multivariate analysis revealed that elevated serum miR-210(OR=2.065,95%CI:1.116～3.821)and TSG-6(OR=1.047,95%CI:1.013～1.083)were independent risk factors for heart failure in DCM patients(P<0.05),whereas higher CTRP3 levels(OR=0.911,95%CI:0.849～0.978)were associated with a protective effect(P<0.05).The sensitivity of serum miR-210,TSG-6,and CTRP3 in predicting heart failure in DCM patients was 72.34%,74.47%,and 74.47%,respectively,with specificities of 62.86%,62.86%,and 68.57%,yielding AUC values of 0.669,0.712,and 0.759,respectively.Conclusions Serum levels of miR-210 and TSG-6 are elevated,whereas CTRP3 levels are reduced in patients with DCM.These biomarkers are closely associated with myocardial fibrosis and cardiac function impairment.Moreover,miR-210,TSG-6,and CTRP3 exhibit significant predictive value for the prognosis of DCM.

Objective To investigate the clinical features of chronic hepatitis C(CHC)patients with hepatitis C virus genotype 3(HCV GT3)infection and the risk factors for disease progression.Methods A multicenter retrospective cohort study was conducted among 1 002 CHC patients from 11 clinical centers in Northwest China from December 2017 to November 2023,and according to their genotype,they were divided into GT1,GT2,GT3,and GT6 groups.Clinical features were compared between the patients with different genotypes.The one-way analysis of variance was used for comparison of normally distributed continuous data between groups,and the Scheffe test was used for further comparison between two groups.The Kruskal-Wallis H test was used for comparison of data with skewed distribution between groups;the chi-square test or Fisher test was used for comparison of categorical data between groups.The multivariate logistic regression analysis was used to explore the influencing factors for the progression of CHC to liver cirrhosis.Results In terms of the genotype,there were 427 patients with GT1 infection,242 with GT2 infection,299 with GT3 infection(210 patients with GT3a infection,87 with GT3b infection,and 2 with unclassified genotype),and 34 with GT6 infection.The patients with GT3 infection had a significantly younger age than those with GT1 infection(51.3±0.5 years vs 53.2±0.6 years,P<0.05)or GT2 infection(51.3±0.5 years vs 53.7±0.8 years,P<0.05),and for the patients with liver cirrhosis,the patients with GT3 infection had a significantly younger age than those with GT1 infection(52.1±0.5 years vs 59.4±0.9 years,P<0.001)or GT2 infection(52.1±0.5 years vs 58.1±1.1 years,P<0.001).Among the patients with GT3 infection,male patients accounted for 77.9%and the patients with liver cirrhosis accounted for 46.2%,which were significantly higher than those among the patients with GT1,GT2 or GT6 infection(all P<0.001).At baseline,the patients with GT3 infection had significantly higher levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)than those with GT1 or GT2 infection,significantly higher aspartate aminotransferase-to-platelet ratio index(APRI)and fibrosis-4(FIB4)than those with GT1,GT2 or GT6 infection,a significantly lower platelet count(PLT)than those with GT2 or GT6 infection,a significantly higher level of alpha-fetoprotein than those with GT2 or GT6 infection,and a significantly lower level of albumin(Alb)than those with GT6 infection(all P<0.05).There were no significant differences between the patients with GT3a infection and those with GT3b infection in age,sex,the proportion of patients with liver cirrhosis,comorbidities,HCV RNA quantification,PLT,ALT,AST,alkaline phosphatase,Alb,APRI,and FIB-4(all P>0.05).The multivariate logistic regression analysis showed that PLT≤150×109/L(odds ratio[OR]=10.72,95%confidence interval[CI]:5.76-35.86,P<0.001)and Alb≤35 g/L(OR=3.74,95%CI:1.22-11.45,P=0.021)were risk factors for liver cirrhosis.Conclusion Most CHC patients with GT3 infection are male in Northwest China,and compared with the patients with other genotypes,such patients tend to have a younger age of onset and higher degrees of liver inflammation activity and fibrosis.Low PLT and a low level of Alb are risk factors for progression to liver cirrhosis in CHC patients with GT3 infection.

Objective:To investigate the prognosis of postoperative adjuvant therapy for hepatocellular carcinoma after conversion therapy of combined targeted therapy and immunotherapy followed by sequential hepatectomy.Methods:The retrospective cohort study was conducted. The clinicopathological data of 103 patients with initially unresectable hepatocellular carcinoma (HCC) who were admitted to 11 medical centers in China, including Mengchao Hepatobiliary Hospital of Fujian Medical University et al, from November 2019 to May 2023 were collected. There were 83 males and 20 females, aged (54±12)years. All 103 patients underwent conversion therapy of tyrosine kinase inhibitors (TKIs) and immune checkpoint inhibitors (ICIs) successfully followed by sequential hepatectomy, of which 72 patients undergoing postoperative adjuvant therapy were divided into the adjuvant therapy group, and 31 patients undergoing postoperative follow-up monitoring were divided into the follow-up monitoring group. Observation indicators: (1) follow-up and postoperative condi-tions; (2) analysis of factors influencing recurrence-free survival time of patients; (3) stratified ana-lysis. Comparison of count data between group was conducted using the chi-square test or Fisher exact probability. The R software was used to draw survival curves, and the Log-rank test was used for survival analysis. Univariate and multivariate analyses were conducted using the Cox proportional hazard model. Results:(1) Follow-up and postoperative conditions. All 103 patients were followed up for 21.0(range, 1.9?47.2)months, with the median recurrence-free survival time of 28.7 months and the 1-, 2-, 3-year recurrence-free survival rates of 68.6%, 55.6%, 41.2%. The median overall survival time of 103 patients was unreached, and the 1-, 2-, 3-year overall survival rates were 90.9%, 82.1%, 69.6%, respectively. The median recurrence-free survival time was 33.1 months in patients of the adjuvant therapy group, with the 1-, 2-year recurrence-free survival rates as 77.2%, 61.5%. The median recurrence-free survival time was 11.1 months in patients of the follow-up monitoring group, with the 1-, 2-year recurrence-free survival rates as 46.6%, 40.8%. There was a significant difference in recurrence-free survival between the two groups of patients ( χ2=5.492, P<0.05). (2) Analysis of factors influencing recurrence-free survival time of patients. Results of multivariate analy-sis showed that pathologic complete response and postoperative adjuvant therapy were independent factors influencing recurrence-free survival time of HCC patients undergoing conversion therapy of combined targeted therapy and immunotherapy followed by sequential hepatectomy ( hazard ratio=0.297, 0.492, 95% confidence interval as 0.137?0.647, 0.268?0.903, P<0.05). (3) Stratified analysis. Of the 71 patients with non-pathologic complete response, the median recurrence-free survival time of 48 patients in the adjuvant therapy group was 24.0 months, with the 1-, 2-year recurrence-free survival rates as 67.4%, 48.8%. The median recurrence-free survival time of 23 patients with non-pathological complete response in the follow-up monitoring group was 7.4 months, with the 1-, 2-year recurrence-free survival rates as 35.0%, 26.3%. There was a significant difference in recurrence-free survival between the 48 patients with non-pathologic complete response in the adjuvant therapy group and the 23 patients with non-pathologic complete response in the follow-up monitoring group ( χ2=5.241, P<0.05). Conclusion:For HCC patients with conversion therapy of TKIs and ICIs followed by sequential hepatectomy, postoperative adjuvant therapy, compared to postoperative follow-up monitoring, can prolong the recurrence-free survival time of patients, of whom cases with non-pathologic complete response can benefit from adjuvant therapy.

Objective To investigate the effects of electroacupuncture at Neiguan(PC6)combined with Buyang Huanwu Decoction on myocardial edema-related proteins and its cardioprotective role in mice with acute high-altitude hypoxic myocardial injury,and to explore the potential mechanisms by which this combined therapy ameliorates acute hypoxic myocardial damage.Methods Mice were randomly divided into control,hypoxia model,electroacupuncture at Neiguan,Buyang Huanwu Decoction,and electroacupuncture at Neiguan+Buyang Huanwu Decoction groups.Except for the normal control group,all other groups were subjected to the establishment of an acute high-altitude hypoxia-induced myocardial injury model.Four days before entering the low-pressure hypoxia animal simulation chamber,the electroacupuncture at Neiguan group was treated with bilateral electroacupuncture at Neiguan,the Buyang Huanwu Decoction group was treated with Buyang Huanwu Decoction by gavage,and the electroacupuncture at Neiguan+Buyang Huanwu Decoction group was treated with a combination of electroacupuncture at Neiguan and Buyang Huanwu Decoction.The intervention lasted for 7 days.The normal control group and the hypoxia model group were handled normally without any other treatment.Myocardial pathology and ultrastructure were evaluated using HE staining and transmission electron microscopy.Serum levels of creatine kinase-MB(CK-MB)and cardiac troponin I(cTn-I)were measured by ELISA.Western blot was performed to quantify β1-AR,cAMP,PKA,and AQP1 protein expression.Results Compared with normal control group,the hypoxia model group exhibited significant myocardial damage,elevated cardiac biomarkers,and upregulated β1-AR/cAMP/PKA pathway proteins with increased AQP1 expression(all P<0.01).The electroacupuncture at Neiguan+Buyang Huanwu Decoction group demonstrated attenuated myocardial injury,reduced biomarker levels,and downregulated target proteins(all P<0.01)versus the hypoxia model group.Conclusion Electroacupuncture at Neiguan combined with Buyang Huanwu Decoction alleviates myocardial edema and injury in acute hypobaric hypoxia by reducing vascular permeability,potentially via suppression of the β1-AR/cAMP/PKA pathway and subsequent inhibition of AQP1 expression.