ObjectiveTo analyze the lipid characteristics and related risk factors of children with Wilson disease （WD）. MethodsThe clinical data of 315 children with WD and 248 healthy children who underwent physical examination were retrospectively collected. The differences in lipid levels between the two groups were compared. The WD group was divided into the normal lipid group （85 cases） and the dyslipidemia group （230 cases） according to lipid levels. General data and biochemical indicators were compared between the two groups， and multivariate Logistic regression analysis was used to analyze the independent risk factors for dyslipidemia in children with WD. ResultsThe incidence of dyslipidemia in children with WD was 73.0% （230/315）， comprising 77 cases （33.5%） of hypertriglyceridemia， 11 cases （4.8%） of hypercholesterolemia， 104 cases （45.2%） of combined hyperlipidemia， 9 cases （3.9%） of hyperlipoprotein（a）emia， and 29 cases （12.6%） of hypoalphalipoproteinemia. There were statistically significant differences in age distribution among different types of dyslipidemia （P<0.05）， whereas no significant differences were observed in gender distribution （P>0.05）. Compared with the normal lipid group， the dyslipidemia group showed significantly higher levels of homocysteine， superoxide dismutase （SOD）， platelet count （PLT）， fibrinogen， alanine aminotransferase， aspartate aminotransferase， γ-glutamyl transferase， uric acid， cystatin C （Cys-C）， type IV collagen （C-IV）， and laminin （all P<0.05）. Multivariate Logistic regression analysis showed that C-IV （OR=5.037， 95% CI： 2.562-9.900）， PLT （OR=2.297， 95% CI： 1.580-3.339）， Cys-C （OR=2.008， 95% CI： 1.395-2.890）， and SOD （OR=1.996， 95% CI： 1.342-2.970） were all independent risk factors for dyslipidemia in children with WD. ConclusionChildren with WD have a high incidence of dyslipidemia， which is dominated by combined hyperlipidemia and hypertriglyceridemia. C-IV， PLT， Cys-C， and SOD are independent risk factors for dyslipidemia in children with WD.

Immunotherapy has recently shown significant potential in treating malignant tumors.However,the immunosuppressive nature of the tumor microenvironment(TME)limits the number of patients who benefit from this approach.Combining ultrasound with drug-loaded microbubble technology can directly induce tumor cell death through cavitation and pore-forming effects;this promotes tumor anti-gen presentation and enhances immune recognition of these antigens.This strategy also modulates the immunosuppressive state of the tu-mor microenvironment,making it more conducive to an effective immune response.It also normalizes blood vessels and reduces interstitial fluid pressure within the tumor,as well as helps therapeutic agents or genes to penetrate tumor tissues,thereby enhancing overall immune efficacy.This review examines the use of low-intensity ultrasound combined with drug-loaded microbubbles to improve the tumor microen-vironment and boost tumor immune responses.

Objective : To explore the risk factors of biliary stricture after liver transplantation and to construct a nomogram prediction model. Methods : The clinical data of 208 liver transplant recipients in hospital were retrospectively analyzed, including 54 cases in the biliary stricture group and 154 cases in the non-biliary stricture group. Multivariate Logistic regression analysis was used to screen out independent predictors, fit the prediction model and construct a visual nomogram to evaluate the prediction model. Survival curves were drawn and multivariate Cox regression analysis was performed. Results : Autoimmune liver diseases(OR=6.610,95%CI: 1.410-30.99), alanine aminotransferase(ALT)(OR=1.007,95%CI: 1.003-1.011), warm ischemia time(OR=1.972,95%CI: 1.399-2.780), cold ischemia time(OR=1.016,95%CI: 1.010-1.022), cytomegalovirus infection(OR=6.037,95%CI: 1.480-24.63) and hepatic vascular stenosis(OR=7.784,95%CI: 2.312-26.20) were independent predictors of biliary stricture after liver transplantation. The area under the curve(AUC) of the nomogram prediction model was 0.921, the cut-off value was 0.238, the sensitivity was 0.889, and the specificity was 0.838. The model showed good discrimination. The Brier score was 0.092, Hosmer-Lemeshow goodness-of-fit testP=0.253, Calibration curve(B=1 000) was in good agreement, and the model showed good calibration. Decision curve analysis(DCA) showed that the application of the model could benefit liver transplant recipients. The postoperative follow-up time was 27-60 months. The cumulative survival rate of the non-biliary stricture group was better than that of the biliary stricture group(P=0.019), but multivariate Cox regression analysis showed that biliary stricture(HR=1.194, 95%CI: 0.624-2.285) was not an independent risk factor for survival after liver transplantation. Conclusion The nomogram model based on autoimmune liver diseases, ALT, warm ischemia time, cold ischemia time, cytomegalovirus infection and hepatic vascular stenosis performs well and can be used to predict the occurrence of biliary stricture after liver transplantation.

Objective To clone and express the heat shock cognate protein 20 (SjHsc20) of Schistosoma japonicum, and to preliminarily investigate its biological characteristics. Methods The target fragment of the SjHsc20 gene was amplified using PCR assay and cloned into the pET-28a(+) expression plasmid to generate the recombinant expression vector pET-28a(+)-SjH-sc20, which was then transformed into Escherichia coli BL21 (DE3) competent cells. The recombinant SjHsc20 (rSjHsc20) protein was induced with isopropyl β-D-thiogalactopyranoside (IPTG) and purified, and the expression of the rSjHsc20 protein was checked with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The immunogenicity of the rSjHsc20 protein was detected using Western blotting, and the transcriptional levels of SjHsc20 were quantified in S. japonicum worms at different developmental stages and in male and female adult worms using real-time quantitative PCR (RT-qPCR) assay. Thirty female BALB/c mice at ages 6 to 8 weeks were divided into three groups, including the rSjHsc20 immunization group, the PBS control group, and the ISA 206 adjuvant group, of 10 mice in each group. Mice in the rSjHsc20 immunization group were subcutaneously immunized with 20 μg rSjHsc20 on days 1, 15 and 31, and animals in the PBS control group were subcutaneously injected with the same volume of PBS on days 1, 15 and 31, while mice in the ISA 206 adjuvant group were subcutaneously immunized with the same volume of ISA 206 adjuvant on days 1, 15 and 31, respectively. All mice in each group were infected with (40 ± 2) S. japonicum cercariae via the abdomen 14 day following the last immunization. Levels of serum specific IgG and its subtypes IgG1 and IgG2 antibodies against rSjHsc20, and the serum titers of anti-rSjHsc20 antibody were detected in mice using indirect enzyme-linked immunosorbent assay (ELISA). All mice were sacrifice 42 days post-infection, and S. japonicum worms were collected from the hepatic portal vein and counted. The eggs per gram (EPG), worm burden reductions and egg burden reductions were estimated to evaluate the protective efficacy of the rSjHsc20 protein. Results The SjHsc20 gene had an open reading frame (ORF) with 756 bp in length and encoded 252 amino acids, and the rSjHsc20 protein had a relative molecular mass of approximately 29 kDa. The rSjHsc20 protein was recognized by the serum of mice infected with S. japonicum and the serum of mice immunized with the rSjHsc20 protein, indicating that rSjHsc20 had a good immunogenicity. There was a significant difference in the transcriptional levels of the SjHsc20 gene among the 7-day (1.001 4 ± 0.065 7), 12-day (2.268 3 ± 0.129 2), 21-day (1.378 5 ± 0.160 4), 28-day (1.196 4 ± 0.244 0), 35-day (1.646 3 ± 0.226 1), 42-day worms of S. japonicum (1.758 0 ± 0.611 1) (F = 38.45, P < 0.000 1), and the transcriptional level of the SjHsc20 gene was higher in the 12-day worms than in worms at other developmental stages (all P values < 0.000 1). The serum levels of anti-rSjHsc20 IgG antibody were 0.106 6 ± 0.010 7, 0.108 3 ± 0.010 4, and 0.553 2 ± 0.069 1 in the PBS control group, ISA 206 adjuvant group, and rSjHsc20 immunization group following the last immunization, respectively, and the serum levels of IgG1 antibody were 0.137 3 ± 0.054 0, 0.181 1 ± 0.096 8, and 1.765 8 ± 0.221 1, while the levels of IgG2a antibody were 0.280 3 ± 0.197 6, 0.274 0 ± 0.146 3, and 1.560 4 ± 0.106 0, respectively. There were significant differences in the serum levels of anti-rSjHsc20 IgG (F = 397.70, P < 0.000 1), IgG1 (F = 401.00, P < 0.000 1) and IgG2a antibodies (F = 229.70, P < 0.000 1) among the three groups, and the serum levels of anti-rSjHsc20 IgG, IgG1 and IgG2a antibodies were higher in the rSjHsc20 immunization group than in the PBS control group and the ISA 206 adjuvant group (all P values < 0.000 1). There was a significant difference in the IgG1/IgG2a ratio among the rSjHsc20 immunization group (1.177 2 ± 0.143 6), the PBS control group (0.428 4 ± 0.199 8) and the ISA 206 adjuvant group (0.559 9 ± 0.181 1) (F = 43.97, P < 0.000 1), and the IgG1/IgG2a ratio was > 1 in the rSjHsc20 immunization group, which was higher than in the PBS control group and the ISA 206 adjuvant group (both P values < 0.000 1). The titers of serum anti-rSjHsc20 antibody were all above 1∶16 384 in the rSjHsc20 immunization group following immunizations on days 1, 15 and 31, indicating that the rSjHsc20 protein had a strong immunogenicity. The mean worm burdens were (16.60±5.75), (15.80±5.58) worms per mouse and (14.40±5.75) worms per mouse in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group 42 days post-infection with S. japonicum cercariae (F = 0.50, P > 0.05), and the EPG were 68 370 ± 22 690, 67 972 ± 19 502, and 41 075 ± 13 251 in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group (F = 4.55, P < 0.05), with lower EPG in the PBS control group and the ISA 206 adjuvant group than in the rSjHsc20 immunization group (both P values < 0.05). Immunization with the rSjHsc20 protein resulted in a worm burden reduction of 13.25% and an egg burden reduction of 39.92% relative to the PBS control group. Conclusions SjHsc20 is successfully cloned and expressed, and the rSjHsc20 protein induces partial immunoprotective effects in mice, which provides a basis for deciphering the biological functions of SjHsc20 and assessing the potential of SjH-sc20 as a vaccine candidate.

Immunotherapy has recently shown significant potential in treating malignant tumors.However,the immunosuppressive nature of the tumor microenvironment(TME)limits the number of patients who benefit from this approach.Combining ultrasound with drug-loaded microbubble technology can directly induce tumor cell death through cavitation and pore-forming effects;this promotes tumor anti-gen presentation and enhances immune recognition of these antigens.This strategy also modulates the immunosuppressive state of the tu-mor microenvironment,making it more conducive to an effective immune response.It also normalizes blood vessels and reduces interstitial fluid pressure within the tumor,as well as helps therapeutic agents or genes to penetrate tumor tissues,thereby enhancing overall immune efficacy.This review examines the use of low-intensity ultrasound combined with drug-loaded microbubbles to improve the tumor microen-vironment and boost tumor immune responses.

Objective:To predict the status of microvascular invasion (MVI) in intrahepatic cholangiocarcinoma (ICC) patients preoperatively based on the radiomics analysis of contrast-enhanced CT to provide imaging evidence for early identification of patients at high risk of recurrence.Methods:Clinical data of 40 ICC patients who underwent radical hepatectomy at Nanjing Drum Tower Hospital from January 2021 to May 2024 were retrospectively collected. Patients were divided into the MVI group ( n=8) and the non-MVI group ( n=32) according to the MVI status of the postoperative pathology report. Whether there were differences in each pathological index between the groups and the efficacy of radiomics analysis of contrast-enhanced CT for the preoperative prediction of MVI were analyzed. The regions of interest (ROI) were outlined on the arterial and venous phase images using the 3D Slicer software. Then, radiomics features were extracted from each ROI based on Python. Finally, the LASSO regression and glm function were used to screen radiomics features and establish a prediction model based on the R language. The established predictive model′s diagnostic efficacy, calibration, and net clinical benefit were evaluated using the receiver operating characteristic (ROC) curve, calibration curve, and decision curve analysis (DCA), respectively. Normally distributed measurement data were expressed as mean±standard deviation ( ± s) and compared using the t-test. Count data were expressed as frequency and compared using the chi-square test. Results:Patients in the MVI group had more poorly differentiated tumors and a significantly higher proportion of lymph node metastases ( P<0.05). The established radiomics prediction model included six features, 1 first-order statistical feature and 5 gray texture features. The area under the ROC curve was 0.87, the sensitivity was 75.0%, and the specificity was 90.6%. The calibration curve showed good agreement between the predicted MVI and actual MVI status, and the decision curve demonstrated that the model could provide a large net clinical benefit. Conclusion:Radiomics analysis of contrast-enhanced CT can identify the MVI status of ICC patients preoperatively and aid in clinical decision-making, providing vital evidence for individualized and precise treatment of ICC.

African swine fever virus(ASFV)I177L gene deletion vaccine is one of the key directions of African swine fever(ASF)live attenuated vaccine research and development.In order to effec-tively distinguish between the wild-type ASFV strain and the I177L gene-deleted strain,specific primers and probes were designed based on ASFV B646L and I177L genes,respectively.After screening and optimization,a duplex real-time PCR method was developed that can simultaneously detect these two genes.The results showed that ASFV B646L and I177L genes were detected spe-cifically and simultaneously by the method developed without cross-reactions with porcine circovir-us type 2,Seneca virus A,classical swine fever virus,foot-and-mouth disease virus,porcine respira-tory and reproductive syndrome virus.The detection limits of the duplex real-time PCR for recom-binant plasmids pUC57-B646L and pUC57-I177L were 1×103 copies/mL.The intra-and inter-as-say coefficients of variation were less than 4％,respectively.Detection of 122 pork and pork prod-ucts using the duplex real-time PCR developed and the real-time PCR recommended by WOAH showed that the coincidence rates of the two methods for B646L gene detection was 100％with two amplification curves appeared in the positive results of the established methods.The method established in this study can be used for the detection of ASFV I177L gene deletion strains,which provides technical support for ASF surveillance and epidemiological investigation.

Objective:To investigate the clinical value of split domino donor auxiliary liver transplantation.Methods:The retrospective and descriptive study was conducted. The clinco-pathological data of 3 liver transplantation recipients who were admitted to Nanjing Drum Tower Hospital affiliated to Nanjing University Medical School and 1 liver transplantation recipient who was admitted to external hospital in September 2018 were collected. The first case was male, aged 22 years, who was diagnosed as type II citrullinemia (CTLN2). The second case undergoing liver transplantation in external hospital was male, aged 59 years, who was diagnosed as decompensated alcoholic cirrhosis. The third case was female, aged 52 years, who was diagnosed as hepatocellular carcinoma of right lobe of liver. The fourth case was female, aged 51 years, who was diagnosed as hepatocellular carcinoma of right lobe of liver. The donor liver from a brain and cardiac death donor was split in vitro into the left liver and the right liver, in which the right liver without middle hepatic vein, and the modified piggyback liver transplantation using the left liver and the classical orthotropic liver transplantation using the right liver was conducted on the first and the second case, respectively. The original liver of the first case was split in vivo into the left liver and the right liver, and the piggyback auxiliary liver transplantation using the left liver and the piggyback auxiliary liver transplantation using the right liver was conducted on the third and the fourth case who underwent extended right hemihepatectomy, respectively. Observation indicators: (1) intraoperative situations; (2) follow-up. Follow-up was conducted using outpatient examination and telephone interview to detect liver function, liver imaging, complication and survival of recipients up to October 2021.Results:(1) Intraoperative situations. Liver transplantation was conducted successfully on the first, third and fourth case, with the operation time, the volume of intraoperative blood loss, the donor liver cold ischemia time, the graft-to-recipient weight ratio were 400 minutes, 370 minutes, 390 minutes, 600 mL, 1 300 mL, 1 600 mL, 230 minutes, 152 minutes, 135 minutes, 1.2%, 0.8%, 1.1%. (2) Follow-up. B-ultrasound examination of the first, third and fourth case after liver transplantation showed that the blood flow was normal, and all the 3 cases discharged and were followed up at postoperative 1, 6 and 12 month. The liver function, the level of blood ammonia and citrulline were normal of the first, third and fourth case at postoperative 1 week. Imaging examina-tion showed normal liver morphology of the first and third case, and a transplanted liver atrophy caused by portal vein steal of the fourth case. ① The level of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil), direct bilirubin (DBil) of the first case before liver transplantation, at postoperative 1 day, 2 day, 3 day, 7 day, 10 day, 6 month and 1 year were 22.8 U/L, 404.1 U/L, 355.5 U/L, 289.6 U/L, 31.0 U/L, 23.1 U/L, 42.1 U/L and 25.8 U/L, 31.5 U/L, 517.7 U/L, 327.6 U/L, 172.9 U/L, 15.9 U/L, 21.4 U/L, 47.5 U/L and 29.7 U/L, 3.8 μmol/L, 92.1 μmol/L, 87.4 μmol/L, 79.7 μmol/L, 90.1 μmol/L, 130.6 μmol/L, 33.8 μmol/L and 25.4 μmol/L, 2.3 μmol/L, 47.0 μmol/L, 44.1 μmol/L, 47.1 μmol/L, 57.4 μmol/L, 70.9 μmol/L, 24.7 μmol/L and 9.7 μmol/L, respectively. The level of citrulline and blood ammonia of the first case before and after liver transplantation were 999.0 μmol/L, 196.0 μmol/L and 14.6 μmol/L, 9.0 μmol/L, respectively. The first case was followed up for 3 years and survived without any liver transplantation related complication. ② The level of ALT, AST, TBil, DBil of the third case before liver transplantation, at postoperative 1 day, 2 day, 3 day, 7 day, 10 day, 6 month and 1 year were 21.3 U/L, 143.9 U/L, 182.0 U/L, 132.0 U/L, 17.2 U/L, 10.1 U/L, 17.6 U/L and 16.8 U/L,20.0 U/L, 291.0 U/L, 227.5 U/L, 106.4 U/L, 15.8 U/L, 10.8 U/L, 17.1 U/L and 19.4 U/L, 6.8 μmol/L, 50.9 μmol/L, 45.0 μmol/L, 34.0 μmol/L, 32.4 μmol/L, 22.3 μmol/L, 12.8 μmol/L and 14.9 μmol/L, 2.5 μmol/L, 18.4 μmol/L, 17.2 μmol/L, 14.9 μmol/L, 14.8 μmol/L, 12.1 μmol/L, 3.6 μmol/L and 4.4 μmol/L. The level of citrulline and blood ammonia of the third case after liver transplantation were 24.9 μmol/L and 16.0 μmol/L. The third case was followed up for 3 years and survived without any liver transplantation related complication. ③ The level of ALT, AST, TBil, DBil of the fourth case before liver transplantation, at postoperative 1 day, 2 day, 3 day, 7 day, 10 day, 6 month and 1 year were 35.0 U/L, 268.7 U/L, 682.0 U/L, 425.8 U/L, 57.5 U/L, 34.0 U/L, 29.4 U/L and 18.1 U/L, 37.0 U/L, 419.1 U/L, 436.2 U/L, 139.5 U/L, 35.2 U/L, 32.4 U/L, 54.7 U/L and 32.8 U/L, 7.1 μmol/L, 64.2 μmol/L, 41.4 μmol/L, 17.6 μmol/L, 34.2 μmol/L, 48.7 μmol/L, 14.1 μmol/L and 21.8 μmol/L, 2.8 μmol/L, 18.9 μmol/L, 16.1 μmol/L, 6.0 μmol/L, 14.6 μmol/L, 26.7 μmol/L, 3.9 μmol/L, 11.8 μmol/L. The level of citrulline and blood ammonia of the fourth case after liver transplantation were 8.4 μmol/L and 47.0 μmol/L. One week after surgery, the transplanted right liver of the fourth case occurred atrophy due to blood stealing from the right branch of the portal vein. B-ultrasound examination showed that the reflux of the hepatic artery and hepatic vein was unobstructed. Immunosuppressants were discontinued 3 months after operation on the fourth case and there was no complication such as rejection, bile leakage, biliary stricture, thrombosis and vascular stricture during follow-up. The fourth case died of lung metastasis 19 months after operation.Conclusion:Split domino donor auxiliary liver transplantation can be used for the treatment of metabolic liver disease and advanced hepatocellular carcinoma.

Objective:The composition of thrombi obtained during mechanical thrombectomy in patients with acute ischemic stroke is analyzed to investigate its relation with stroke etiology and its influence in surgical parameters and clinical prognoses.Methods:The thrombi and clinical data of 41 patients with acute ischemic stroke directly treated by mechanical thrombectomy in our hospital from January 2019 to December 2019 were collected. Hematoxylin-eosin (HE) staining was used to quantitatively analyze the composition of thrombi, and the components of thrombi in patients with different causes of stroke (large artery atherosclerosis [LAA], cardiogenic embolism [CE], and unexplained type) were compared. These patients were divided into erythrocyte-rich group (erythrocyte content>fibrin content) and fibrin-rich group (erythrocyte content0.05). Conclusion:Thrombus composition has influence in surgical parameters, but no influence in clinical prognoses.

Objective To explore the value of intravoxel incoherent motion (IVIM) analysis in evaluation of pathological complete response (pCR) in patients with locally advanced rectal cancer (LARC) after neoadjuvant chemotherapy.Methods Totally 61 patients with pathologically proven rectal adenocarcinoma were retrospectively analyzed.All patients underwent MR scan,including multi-b DWI IVIM sequence scanning before and after neoadjuvant chemotherapy.ADC values and IVIM parameters (slow diffusion coefficient [D],fast diffusion coefficient [D*] and perfusion fraction [f]) were measured by two radiologists independently.Then the patients were divided into pCR group (n=14) and non-pCR group (n=47).Independent-samples t test was used to compare all the parameters before and after therapy,as well as the percentage changes (△ratio) between pCR and non-pCR group.Area under the curve (AUC) was used to evaluate the diagnostic performance.Results ADC (P=0.002) and D (P=0.007) values were significantly lower before therapy,while △ratio ADC (P＜0.001) and △ratio D (P =0.002) were significantly higher in pCR group compared with those in non-pCR group.D*,f and their △ratio value were not statistically different pre-chemotherapy and post-chemotherapy in both two groups (all P＞0.05).△ratio D had the highest AUC (0.786,P＜0.01).Conclusion IVIM-derived D values can help to predict and differentiate pCR of LARC after neoadjuvant chemotherapy.