Background/Aims: Liver transplantation (LT) following total hepatectomy is a life-saving treatment for hepatocellular carcinoma (HCC). The HCC recurrence after LT hinders the effectiveness of the procedure. The objective of this study is to develop a pre-operative risk stratification model based on a liquid biopsy. Methods: We conducted a comprehensive multi-omics study of 260 HCC patients from three centers, including clinical data, low-coverage whole-genome sequencing of cell-free DNA (cfDNA) from plasma, as well as whole-exome, single-nucleus RNA, and spatial transcriptomics from matched tumor and non-tumor tissues. Results: We identified cfDNA-derived copy number alteration (CNA) signatures associated with post-transplant recurrence. By integrating cfDNA-derived CNA profiles with single-cell transcriptomic data, we traced recurrence-associated cfDNA to a distinct subpopulation of malignant cells within the primary tumor. These cells were embedded in a pro-metastatic microenvironment of specialized endothelial subtypes and cancer-associated fibroblasts. Notably, most recurrence-associated lesions were detectable in cfDNA prior to liver transplantation (LT). Building on these insights, we developed the ZJU Criteria based on CNA fragments and tumor markers, a pre-LT risk prediction tool that integrates conventional clinical factors with cfDNA-derived CNA signatures, and validated it using internal and independent external cohorts. Conclusion Our findings suggest that post-transplant recurrence commonly originates from advanced subclones that emerge late during tumor evolution. The ZJU Criteria provides an accurate, non-invasive strategy that significantly improves pre-LT risk stratification and clinical decision-making for patients with HCC.

Objective To review the diagnostic information of 3368 viral hepatitis B inpatient cases reported by some healthcare institutions in Qinghai Province in the National Notifiable Disease Report System(NNDRS)in 2021,and to analyse the consistency of their reports and influence factor.Methods The study used the typical survey method to study 3368 hospitalized cases of hepatitis B reported by 32 medical institutions in Qinghai Province in 2021 from July 2022 to April 2023.The consistency between the reported classification and the reviewed classification was evaluated by reviewing diagnostic information of the cases,combined with the Kappa consistency test,and the consistency-influencing factors were analysed in a univariate and multivariate analysis.Results The consistency between acute,chronic,and unclassified reporting classifications and review classifications was poor(Kappa＜0.40);the consistency between acute and chronic review was poor(Kappa＜0.40).The main influence factor of the consistency of review included the level of medical institution,the patient's gender,occupation,past history,and the unit of diagnosis of initial onset of illness.Provincial medical institutions reported higher consistency,and male cases and herder cases had better consistency than female cases and other occupations.Review consistency of cases with past history was better than that of cases with unclear past history.Conclusion The consistency of diagnosis and reporting of hepatitis B cases in healthcare institutions in Qinghai Province in 2021 was low,with problems of inaccurate classification and duplicate reporting.It is recommended to strengthen the training and supervision of healthcare institutions,standardise the filling of'attached card'information,and promote the hepatitis B core antibody IgM test to improve the classification and diagnosis of acute and chronic hepatitis B,so as to comprehensively improve the accuracy of the diagnosis and reporting of hepatitis B cases.

BACKGROUND:Studies have shown that intradiscal injection of syringin solution can improve the structure and function of the intervertebral disc,prevent and slow down the process of intervertebral disc degeneration in rats.However,the biological half-life of syringin is short and it is difficult for it to continue to play a role in the intervertebral disc.Its bioavailability needs to be further improved.OBJECTIVE:To observe the effect of syringin-chitosan hydrogel on intervertebral disc degeneration in rats and the mechanism of syringin in the treatment of intervertebral disc degeneration.METHODS:(1)Cell experiment:Passages 2-5 rat nucleus pulposus cells were divided into four groups for treatment.The normal control group did not undergo any treatment.The degeneration group was added with interleukin-1β(to establish the intervertebral disc degeneration cell model).The drug group was added with interleukin-1β and syringing.The inhibitor group was added with interleukin-1β,syringing,and phosphatidylinositol 3-kinase(PI3K)inhibitor LY294002.After 24 hours of treatment,apoptosis,extracellular matrix,oxidative stress,and apoptosis-related proteins and PI3K/protein kinase B(AKT)signaling pathway proteins were detected respectively.(2)Animal experiment:Syringin-chitosan hydrogels were prepared,and the micromorphology and slow-release properties of the hydrogels were tested.Thirty SD rats were randomly divided into model control group,model intervention group,hydrogel group,syringin solution group,and syringin hydrogel group,with 6 rats in each group.The intervertebral disc degeneration model was established by the acupuncture method.Immediately after model establishment,the rats in model intervention group,hydrogel group,syringin solution group,and syringin hydrogel group were injected with PBS,chitosan hydrogel,syringin solution,and syringin-chitosan hydrogel,respectively.The samples were taken 8 weeks after modeling for histological detection.RESULTS AND CONCLUSION:(1)Cell experiment:Compared with the normal control group,apoptosis rate,reactive oxygen species level,and expression of BAX,cleaved caspase-9,cleaved caspase-3,and matrix metalloproteinase 13 protein were increased in the nucleus pulpocytes in the degeneration group(P＜0.05),and the expression levels of p-PI3K,p-AKT,BCL-2,and type Ⅱ collagen were decreased(P＜0.05).Superoxide dismutase activity decreased(P＜0.05).Compared with the degeneration group,apoptotic rate,reactive oxygen species level,and expression of BAX,cleaved caspase-9,cleaved caspase-3,and matrix metalloproteinase 13 protein were decreased in the syringin solution and syringin solution groups(P＜0.05),and expression levels of p-PI3K,p-AKT,BCL-2,and type Ⅱ collagen were increased(P＜0.05).Superoxide dismutase activity increased(P＜0.05).LY294002 could partially inhibit the effect of syringin.(2)Animal experiment:Syringin-chitosan hydrogel had a loose porous structure and good slow-release performance.Hematoxylin-eosin and safranin O-fast green staining showed that compared with the model control group and model intervention group,chitosan hydrogel,syringin solution and syringing-chitosan hydrogel could improve the intervertebral disc degeneration in different degrees,and the therapeutic effect of syringing-chitosan hydrogel was better than that of hydrogel and drug solution alone.(3)These findings indicate that syringin can regulate apoptosis of nucleus pulposus cells and extracellular matrix degradation induced by oxidative stress by activating PI3K/AKT signaling pathway,thus delaying disc degeneration.Compared with syringin injection alone,syringin loading in chitosan hydrogel can further delay the progression of intervertebral disc degeneration in rats.

Objective To investigate the application of targeted next-generation sequencing test(tNGS)in detecting patho-gens and drug resistance genes in the diagnosis and treatment of Mycoplasma pneumoniae pneumonia(MPP)in chil-dren.Methods The clinical data of children diagnosed with MPP and hospitalized in Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology from July 2022 to December 2023 were analyzed.The differences of MP se-quence detected by tNGS in children of different gender,age,course,and clinical features were compared.The performance of tNGS in detecting macrolides resistance genes was assessed.The optimal prediction cut-off value of MP resistance gene was ex-plored by ROC.Results A total of 497 cases were included,including 257(51.7％)male and 240(48.3％)female.The age dis-tribution was from 3 months to 13 years old[(6.08±2.68)years old in average],and the average durations of fever and cough were(6.04±3.42)and(7.46±5.54)days,respectively.MP was detected in all patients by tNGS method.There was no signifi-cant difference in different gender or age groups.The MP sequence count in patients with positive nucleic acid was significantly higher than that in patients with negative nucleic acid.The MP sequence count in lower respiratory specimens was significantly higher than that in upper respiratory specimens.MP sequence count in severe cases was significantly higher than that in non-se-vere cases,but there was no significant difference between patients with or without pulmonary complications and refractory or non-refractory cases.The overall diagnostic sensitivity of tNGS for detecting MP drug-resistant genes was 92.8％,specificity was 27.8％,PPV was 48.3％,NPV was 84.2％,and the mutation site was 23S rRNA:A2063G.The ratio of MP pathogenic se-quence to drug-resistant gene sequence was 6.83,which was the best cut-off value for predicting drug resistance.The negative predictive value was higher when the MP pathogenic sequence ratio was greater than 6.83.Conclusion tNGS has the value in early diagnosis of MPP.The MP sequence counts may have a guiding value in predicting disease severity.The negative predictive value is higher in drug-resistant gene detection.

Objective:To identify the risk factors associated with conversion to cesarean delivery in parturients receiving epidural labor analgesia and to construct a predictive model.Methods:In this prospective cohort study, 718 parturients with a singleton fetus at 37-42 weeks of gestation age, aged 22-45 yr, of American Society of Anesthesiologists Physical Status classification Ⅱ, who received epidural labor analgesia at the Women′s Hospital affiliated to Zhejiang University School of Medicine between November 2022 and August 2023, were included and divided into 2 groups based on the final mode of delivery: cesarean delivery group (CD group) and vaginal delivery group (VD group). Data collected included maternal age, height, body mass index at labor onset, gestational age, pregnancy complications, mode of conception, parity, premature rupture of membranes, mode of labor onset and details of labor analgesia. Risk factors for conversion to cesarean delivery were analyzed using multivariate logistic regression analysis, and a prediction model was developed. The receiver operating characteristic curve was ploted, and the area under the receiver operating characteristic curve was calculated to evaluate the accuracy of the predictive model.Results:A total of 682 parturients were included in the final statistical analysis, with 103 in CD group and 579 in VD group. Multivariate logistic regression analysis showed that maternal age≥35 yr, height<160 cm, body mass index at labor onset ≥28 kg/m 2, nulliparous parturients, the use of Foley balloon catheter or dinoprostone suppositories for cervical ripening were risk factors for cesarean delivery, and cervical dilation during analgesia was a protective factor. The area under the receiver operating characteristic curve for the predictive model constructed based on the aforementioned influencing factors yielded predicting conversion to cesarean delivery due to failed vaginal trial of labor was 0.791 (95% confidence interval 0.744-0.837, P<0.001), with a sensitivity of 73.6% and a specificity of 70.9%. Conclusions:Nulliparous parturients, maternal age≥35 yr, height<160 cm, body mass index at labor onset≥28 kg/m 2, and the use of Foley balloon catheter or dinoprostone suppositories for cervical ripening are risk factors for cesarean delivery, and cervical dilation during analgesia is a protective factor. The predictive model established based on these influencing factors has high predictive performance.

Respiratory syncytial virus (RSV) is the most common respiratory pathogen in infants and young children, which can lead to severe pneumonia and even death. Early diagnosis and intervention are crucial for reducing complications and sequelae and improving prognosis. This study presents the complete diagonsis and treatment process of a typical severe RSV pneumonia patient. For high-risk children, the condition can progress rapidly after RSV infection, and may present with respiratory failure and multiple extrapulmonary complications, such as heart failure, liver and kidney function impairment, coagulation dysfunction. After active symptomatic and supportive treatment, the prognosis is generally good. RSV infection will not lead to long-lasting immunity, but as the child grows older and their immune function gradually improves, the condition of a secondary infection is generally relatively mild. Currently, there are no specific antiviral drugs or safe and effective vaccines for RSV. Long-acting monoclonal antibodies are one of the effective means to prevent RSV infection in infants and young children and have been approved for use in China.

Objective::This study aimed to elucidate the effect of the lipopolysaccharides/toll-like receptor 4 (LPS/TLR4) pathway on early atherosclerosis (AS) development and its associated changes in fecal metabolites, thereby providing an experimental foundation for strategies to prevent and treat early AS.Methods::Twelve Tibetan miniature pigs aged 4-5 months were divided into normal control (NC) group and AS group (6 pigs in each). The group assignment was primarily based on body weight; Secondary criteria, including glucose, lipid profiles, and inflammatory indices, were considered to ensure balanced baseline characteristics between the 2 groups (all P > 0.05). AS group received a high-fat diet for 16 weeks to establish an AS model, while the NC group received a normal diet. Subsequently, serum levels of lipids and various inflammation and oxidative stress markers were measured. Pathological changes in the aorta and colon tissue, LPS/TLR4 pathway-associated protein expressions in the aorta, as well as occludin and zonula occludens-1 in the colon were also assessed. Proton nuclear magnetic resonance spectra technology was employed for the metabolomic analysis of fecal extracts. Results::The lipid metabolism was disrupted in AS group, with significantly higher total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol levels ((12.24 ± 5.24) mmol/L vs. (1.86 ± 0.27) mmol/L, P = 0.004,6; (2.39 ± 0.50) mmol/L vs. (0.83 ± 0.07) mmol/L, P = 0.000,5; (6.94 ± 2.87) mmol/L vs. (0.77 ± 0.18) mmol/L, P = 0.003,3), as compared to that in NC group. Serum factors, including LPS, tumor necrosis factor-α, and malondialdehyde levels of AS group were significantly higher than that of NC group ((1,230.00 ± 192.70) EU/L vs. (695.70 ± 213.70) EU/L), P = 0.001,1; (424.20 ± 176.90) ng/L vs. (51.20 ± 26.61) ng/L, P = 0.023,5; (3.60 ± 0.77) nmol/mL vs. (2.62 ± 0.21) nmol/mL, P = 0.025,4). Pathological evaluations revealed prominent lipid deposition area in the aortic arch, thoracic aorta, and abdominal aorta of the AS group compared with that of the NC group (4.17% ± 2.30% vs. 0, P = 0.006,7; 6.23% ± 2.95% vs. 0, P = 0.003,6; 3.78% ± 2.18% vs. 0, P = 0.008,1). TLR4, nuclear factor kappa-B p65, and tumor necrosis factor-α expression in the aorta tissue of the AS group were upregulated, whereas occludin and zonula occludens-1 expression in colon tissues was downregulated. Additionally, metabolomics identified significant differences in 21 metabolites in the feces of the AS group compared to the NC group, with further analysis linking these differences to amino acid metabolism. Conclusions::The Tibetan miniature pig model of early AS induced by high-fat intake displayed pronounced chronic inflammation. Preliminary findings suggest that the underlying mechanisms may be associated with the LPS/TLR4 pathway and intestinal metabolic disorders.

Objective:To explore the healing effects of internal heat needle and electroacupuncture on post-stroke shoulder pain and their impacts on serum interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α).Methods:From January 2023 to March 2025, 94 patients with post-stroke shoulder pain admitted to the Affiliated Hospital of Hangzhou Normal University were included and separated into two groups according to a random number table. The 47 cases in the electroacupuncture group adopted electroacupuncture, while 47 cases in the internal heat needle group adopted internal heat needle. The pre- and post-treatment limb function, quality of life, healing effect, visual analogue scale (VAS), and serum IL-6, IL-1β, and TNF-α were compared.Results:After treatment, the Constant-Murley Shoulder Scale (CMS) score, shoulder range of motion (flexion, outward rotation, extension), Fugl-Meyer Assessment-Upper Extremity (FMA-UE) score, and Modified Barthel Index (MBI) in the internal heat acupuncture group were all better than those in the electroacupuncture group:(63.45 ± 10.31) points vs. (56.51 ± 9.15) points, (136.67 ± 32.44)° vs. (109.32 ± 25.01)°, (51.64 ± 12.84)° vs. (42.95 ± 11.72)°, (34.35 ± 10.15)° vs. (29.53 ± 9.82)°, (45.63 ± 10.25) points vs. (39.16 ± 10.43) points, (83.42 ± 16.09) points vs. (74.55 ± 15.83) points, there were statistical differences ( P<0.05). While the levels of serum IL-1β, IL-6, TNF-α, and VAS score in the internal heat needle group were decreased in the electroacupuncture group: (7.39 ± 3.68) ng/L vs. (9.28 ± 4.10) ng/L, (19.25 ± 2.12) ng/L vs. (28.50 ± 2.43) ng/L, (9.55 ± 3.54) ng/L vs. (11.17 ± 3.77) ng/L,(1.69 ± 0.45) points vs. (2.13 ± 0.56) points, there were statistical differences ( P<0.05). The therapeutic effect in the internal heat needle group was significantly higher than that in the electroacupuncture group:93.62% (44/47) vs. 78.72% (37/47), there was statistical difference ( P<0.05). Conclusions:For post-stroke shoulder pain, internal heat needle therapy has a clear healing effect than electroacupuncture, and can effectively reduce the levels of inflammatory factors such as IL-1β, IL-6, and TNF-α in serum.

Objective:To evaluate long-term outcomes of endoscopic papillectomy (EP) for duodenal papillary adenomas and to identify risk factors for incomplete resection.Methods:Clinical data of 180 patients diagnosed as having duodenal papillary adenoma via postoperative pathology after EP in Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School from January 2010 to December 2022 were retrospectively analyzed. Patients were divided into two groups based on their postoperative margin status: the complete resection group (negative resection margins) and the incomplete resection group (positive/uncertain resection margins). Recurrence rates were compared between the two groups, and logistic regression analysis was performed to identify risk factors for incomplete resection.Results:Among the 180 patients included in the study, 137 underwent complete resection, and 43 had incomplete resections. Recurrence rate was significantly higher in the incomplete resection group than that in the complete resection group (30.2% VS 15.3%, χ2=4.75, P=0.029). logistic regression analysis indicated that high-grade intraepithelial neoplasia was an independent risk factor for incomplete resection ( OR=2.43, 95% CI:1.12-5.26, P=0.024). Conclusion:Patients with incomplete resection after EP have a higher recurrence rate in the long-term follow-up. High-grade intraepithelial neoplasia is an independent risk factor for incomplete resection. Close surveillance and aggressive management are warranted for patients with positive or uncertain resection margins to mitigate the recurrence risk.

BACKGROUND:Studies have shown that intradiscal injection of syringin solution can improve the structure and function of the intervertebral disc,prevent and slow down the process of intervertebral disc degeneration in rats.However,the biological half-life of syringin is short and it is difficult for it to continue to play a role in the intervertebral disc.Its bioavailability needs to be further improved.OBJECTIVE:To observe the effect of syringin-chitosan hydrogel on intervertebral disc degeneration in rats and the mechanism of syringin in the treatment of intervertebral disc degeneration.METHODS:(1)Cell experiment:Passages 2-5 rat nucleus pulposus cells were divided into four groups for treatment.The normal control group did not undergo any treatment.The degeneration group was added with interleukin-1β(to establish the intervertebral disc degeneration cell model).The drug group was added with interleukin-1β and syringing.The inhibitor group was added with interleukin-1β,syringing,and phosphatidylinositol 3-kinase(PI3K)inhibitor LY294002.After 24 hours of treatment,apoptosis,extracellular matrix,oxidative stress,and apoptosis-related proteins and PI3K/protein kinase B(AKT)signaling pathway proteins were detected respectively.(2)Animal experiment:Syringin-chitosan hydrogels were prepared,and the micromorphology and slow-release properties of the hydrogels were tested.Thirty SD rats were randomly divided into model control group,model intervention group,hydrogel group,syringin solution group,and syringin hydrogel group,with 6 rats in each group.The intervertebral disc degeneration model was established by the acupuncture method.Immediately after model establishment,the rats in model intervention group,hydrogel group,syringin solution group,and syringin hydrogel group were injected with PBS,chitosan hydrogel,syringin solution,and syringin-chitosan hydrogel,respectively.The samples were taken 8 weeks after modeling for histological detection.RESULTS AND CONCLUSION:(1)Cell experiment:Compared with the normal control group,apoptosis rate,reactive oxygen species level,and expression of BAX,cleaved caspase-9,cleaved caspase-3,and matrix metalloproteinase 13 protein were increased in the nucleus pulpocytes in the degeneration group(P＜0.05),and the expression levels of p-PI3K,p-AKT,BCL-2,and type Ⅱ collagen were decreased(P＜0.05).Superoxide dismutase activity decreased(P＜0.05).Compared with the degeneration group,apoptotic rate,reactive oxygen species level,and expression of BAX,cleaved caspase-9,cleaved caspase-3,and matrix metalloproteinase 13 protein were decreased in the syringin solution and syringin solution groups(P＜0.05),and expression levels of p-PI3K,p-AKT,BCL-2,and type Ⅱ collagen were increased(P＜0.05).Superoxide dismutase activity increased(P＜0.05).LY294002 could partially inhibit the effect of syringin.(2)Animal experiment:Syringin-chitosan hydrogel had a loose porous structure and good slow-release performance.Hematoxylin-eosin and safranin O-fast green staining showed that compared with the model control group and model intervention group,chitosan hydrogel,syringin solution and syringing-chitosan hydrogel could improve the intervertebral disc degeneration in different degrees,and the therapeutic effect of syringing-chitosan hydrogel was better than that of hydrogel and drug solution alone.(3)These findings indicate that syringin can regulate apoptosis of nucleus pulposus cells and extracellular matrix degradation induced by oxidative stress by activating PI3K/AKT signaling pathway,thus delaying disc degeneration.Compared with syringin injection alone,syringin loading in chitosan hydrogel can further delay the progression of intervertebral disc degeneration in rats.