Objective To optimize the extraction, separation and purification process of Actinoside E. Methods Single factor experiment combined with orthogonal test was used to determine the optimal extraction process of Actinoside E using its content as an index. The extracts were separated and purified by optimizing the chromatographic conditions of macroporous resin, silica gel and ODS column. Results 25 times amount of 55% ethanol with heating reflux at 95℃ for one hour were used as the optimal extraction process of Actinoside E. The optimum separation and purification process was as follows: D101 macroporous resin column was eluted with 7 BV of 50% ethanol, silica gel column was eluted with 5 BV of ethyl acetate-ethanol（10∶1）and 50% methanol eluted fraction was purified repeatedly by ODS column to obtain Actinoside E. The transfer rate of Actinoside E in the whole process was 53.70%, the yield was 0.35%, and the purity was 99.9%. Conclusion The process is stable and viable, which can provide material foundation for the development and utilization of Actinoside E.

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.

ObjectiveTo investigate the mutation status of the MET gene in tumor patients， with a focus on its expression in lung cancer tissues and its impact on patient prognosis， providing theoretical basis and potential targets for precise diagnosis and treatment of the tumors. MethodsA total of 1，627 tumor patient samples were collected from January 2021 to September 2025. High-throughput sequencing technology was used to conduct DNA-level gene detection on 137 tumor patients with MET mutations. The distribution of mutation sites in exons and introns， as well as the occurrence frequencies of MET gene amplification and gene fusion were analyzed， with particular attention paid to the mutation frequencies of each exon and the high-frequency variant types. Immunohistochemical staining was performed to analyze the expression levels of c-MET protein in tumor tissues of the 137 patients. Immunohistochemical analysis was conducted on tumor tissues of 9 patients with MET gene exon 14 skipping mutations to study the association between c-MET protein expression and this specific mutation type. RNA sequencing results， immunohistochemical staining data， and follow-up prognosis of lung cancer from the GEPIA2 database and TCGA database were comprehensively analyzed to explore the potential relationship between MET gene expression in tumor tissues and patient prognosis. ResultsAmong 1，627 tumor patients， 137 MET mutation patients were detected， including 93 males with an average age of （66.85±9.61） years and 44 females with an average age of （59.43±11.08） years. Lung cancer was the most common tumor type （117 cases）. MET gene expression was lower in breast cancer and glioblastoma multiforme， but higher in lung cancer and colorectal cancer as compared with the expression in normal tissues. Survival analysis showed that patients with high MET gene expression in lung adenocarcinoma had a shorter overall survival time than those with low expression. A total of 961 mutations were detected by next-generation sequencing， including 547 exon mutations and 374 intron mutations. MET gene amplification was found in 13 cases and gene fusion in 27 cases. The highest mutation frequency was in exon 2， followed by exon 5， exon 4， and exon 19. High-frequency variant types included c.2890C>A （p.L964M） on exon 4 and c.3028G>T （p.D1010Y） on exon 5. The c-MET protein expression was closely related to the type of gene mutation. Missense mutations often led to positive or strongly positive c-MET protein expression， while nonsense mutations often resulted in weak or absent c-MET protein expression. Among the 9 patients with MET gene exon 14 skipping mutations， the c-MET protein expression score was mostly 0 - 1， with only 1 case scoring 2. Among the 117 lung cancer patients， the incidence rate was higher in males than in females， and more common among stage Ⅲ-Ⅳ patients than among stage I-Ⅱ patients. Out of the 41 patients treated with surgery + targeted drugs （gutemitinib， savolitinib）， 8 cases of recurrence or metastasis were found during follow-up. ConclusionMET gene mutations exhibit specific mutation characteristics in different tumor types， and its expression levels show significant differences between various tumor tissues and normal tissues. It is closely related to the poor prognosis of lung adenocarcinoma patients. We found that c-MET protein expression is closely related to the type of MET gene mutation， especially in patients with MET gene exon 14 skipping mutations， which presents a characteristic expression pattern. This provides an important basis for the preliminary screening using immunohistochemistry in clinical practice.

Objective:To explore the prognostic value of peripheral blood lymphocyte subsets in patients with newly diagnosed multiple myeloma (NDMM) .Methods:The study retrospectively analyzed 133 patients with NDMM admitted to the General Hospital of Tianjin Medical University General Hospital between 2017 and 2022. The least absolute shrinkage and selection operator (LASSO) regression was used to screen the predictive subgroups from the peripheral blood lymphocyte subsets, and the optimal cutoff value was calculated through receiver operating characteristic curve analysis. A nomogram was constructed based on the results of the multiple-factor analysis, and the predictive performance of the nomogram was evaluated by the concordance index and calibration curve. Kaplan-Meier curves and log-rank tests were conducted to compare the differences in overall survival (OS) and progression-free survival between the high-risk and low-risk immune risk scores groups.Results:Using LASSO regression, the percentages and absolute counts of CD16 +CD56 + NK cells, CD3 + T lymphocytes, CD3 +CD8 + T lymphocytes, and CD3 -CD19 + B lymphocytes were selected as predictive subgroups. The immune risk score of patients with NDMM was calculated based on the coefficients of each lymphocyte subgroup. The area under the curve of the immune risk score was 0.737, and the optimal cutoff value was -1.834. Based on this, the patients were divided into high-risk and low-risk groups. Survival analysis showed a significant difference in the 3-year OS rate between the high-risk and low-risk immune risk score groups (87.4% vs 49.0%, P<0.001), and a significant difference in the 3-year OS rate between the high-risk and low-risk immune risk score groups in patients with minimal residual disease negative (100% vs 68.6%, P=0.001). Multivariate analysis showed that serum calcium ( P=0.034), high-risk cytogenetic abnormalities ( P=0.002), and immune risk score ( P<0.001) were prognostic factors for patients with NDMM, and a nomogram was constructed based on these factors. The consistency index of the nomogram was 0.793, and the calibration curve showed good predictive ability. The nomogram can accurately classify the risk of different prognostic staging systems. Conclusions:The combined analysis of lymphocyte subsets in the peripheral blood has an important value in predicting the prognosis of patients with NDMM.

AIM To investigate the effects of Qifu Yixin Granules on cardiac inflammation in a rat model of heart failure.METHODS The rats were induced into chronic heart failure(CHF)models by 6-week intraperitoneal injection of doxorubicin followed by the random assignment of the successful rat models into the model group,the captopril group(22.5 mg/kg),and the low-dose,medium-dose,and high-dose Qifu Yixin Granules groups(2.84,5.67,11.34 g/kg),in contrast to the normal rats of the blank group.The rats had their body weight monitored;their cardiac function assessed by echocardiography;their serum levels of NT-proBNP,TNF-α,IL-6,IL-1 and CRP measured by ELISA;their cardiac morphological alterations observed by HE and Masson staining;their cardiac protein expressions of TLR4,MyD88 and NF-κB detected by immunohistochemistry and Western blot;and their cardiac mRNA expressions of TLR4,MyD88 and NF-κB measured by RT-qPCR.RESULTS Compared to the blank group,the model group exhibited significantly reduced body weight,LVEF and LVFS(P＜0.01),alongside significantly elevated LVEDD,LVESD,and serum concentrations of NT-proBNP,TNF-α,IL-6,IL-1 and CRP(P＜0.01).Additionally,the model group displayed greater myocardial inflammatory cell aggregation,increased collagen deposition(P＜0.01);and upregulated myocardial protein and mRNA expressions of TLR4,MyD88 and NF-κB(P＜0.01).Compared to the model group,the groups intervened with captopril or medium/high dose Qifu Yixin Granules demonstrated significantly increased body weight,LVEF and LVFS(P＜0.05,P＜0.01);significantly reduced LVEDD,LVESD,and serum levels of the aforementioned indicators(P＜0.05,P＜0.01);mitigated inflammation and collagen deposition(P＜0.05,P＜0.01);and downregulated myocardial protein and mRNA expressions of TLR4,MyD88 and NF-κB(P＜0.05,P＜0.01).CONCLUSION Qifu Yixin Granules attenuate cardiac inflammation and improve cardiac function in doxorubicin-induced CHF rats;this therapeutic effect is mediated by inhibiting the activation of the TLR4/MyD88/NF-κB signaling pathway.

Objective:To evaluate the efficacy and safety of percutaneous nephrolithotomy (PCNL) in the treatment of severe acute pancreatitis (SAP) with infected pancreatic necrosis (IPN).Methods:A retrospective analysis was conducted on the clinical data of 113 patients with SAP and IPN admitted to Qingdao Municipal Hospital between January 2018 and June 2023. The cohort included 66 males and 47 females, aged (46.2±13.6) years. Based on the treatment approach, patients were divided into two groups: those who underwent percutaneous catheter drainage (PCD) alone were assigned to the PCD group ( n=60), and those who received PCNL were assigned to the PCNL group ( n=53). Clinical parameters such as age, sex, number of debridement procedures, and postoperative complications were recorded. Results:Compared with the PCD group, the PCNL group had a higher CT severity index [8(6, 9) vs. 7(6, 8)], a greater proportion of patients with multiloculated abscesses [71.7%(38/53) vs. 41.7%(25/60)], and a higher percentage of patients with IPN involving >50% necrosis [56.6%(30/53) vs. 33.3%(20/60)]. These differences were statistically significant (all P<0.05). The PCNL group also showed a higher proportion of patients with post-treatment IPN necrosis involving <30% [91.4%(49/53) vs. 70.8%(42/60)], a lower rate of requiring step-up laparoscopic or open surgery [3.8%(2/53) vs. 10.0%(6/60)], fewer debridement sessions [2(1, 4) vs. 4(2, 6)], and a shorter total hospital stay [45(35, 58) d vs. 54(23, 72) d]. These differences were statistically significant (all P<0.05). Additionally, the PCNL group had lower rates of postoperative complications, including enteric fistula [3.8%(2/53) vs. 11.7%(7/60)], intra-abdominal bleeding [5.7%(3/53) vs. 13.3%(8/60)], and pancreatic fistula [15.1%(8/53) vs. 20.0% (12/60)]. These differences were also statistically significant (all P<0.05). The recurrence rate of abscesses was significantly lower in the PCNL group [11.3%(6/53) vs. 16.7%(10/60), χ2=4.14, P=0.042]. Conclusion:PCNL is an effective treatment for SAP complicated by IPN. Compared with PCD, it improves the clearance of necrotic tissue, reduces the number of debridement procedures, shortens the total hospital stay, and lowers the risk of postoperative complications.

Objective:To analyze the clinical efficacy of asymmetric T 1 transpedicular wedge resection Smith-Petersen osteotomy (T 1 SPO) in the treatment of stiff cervical thoracic lateral kyphosis deformity. Methods:This is a retrospective case series study. The clinical data of nine patients with stiff cervical thoracic kyphosis who underwent asymmetric T 1 SPO corrective treatment from June 2012 to October 2022 were collected. There were 7 males and 2 females, aged 45 to 68 years. The surgery time, intraoperative blood loss, and complications were recorded. The chin brow vertical angle (CBVA), cervical thoracic kyphosis Cobb angle, cervical thoracic scoliosis Cobb angle, and cervical thoracic sagittal axis (C 2-T 1 sagittal vertical axis, SVA) before surgery, after surgery, and at the last follow-up were measured and the correction rates were calculate. Results:All 9 patients successfully completed the surgery. The operation time ranged from 245 to 320 minutes, and the intraoperative blood loss was 1 400 to 2 200 ml. All patients were followed up for 24 to 48 months. The preoperative CBVA was 93.7° to 112.0°, which improved to 25.2° to 31.7° at the last follow-up, with an correction rate of 73.4%. The preoperative cervicothoracic kyphosis Cobb angle was -57.0° to -16.6°, which improved to 10.3° to 18.5° at the last follow-up, with an correction rate of 166.7%. The preoperative scoliosis Cobb angle was 13.0° to 16.5°, which improved to 2.2° to 3.8° at the last follow-up, with an correction rate of 84.9%. The preoperative SVA was 7.8 to 12.5 cm, which improved to 4.5 to 6.8 cm at the last follow-up, with an correction rate of 42.3%. One patient experienced numbness and weakness in the left hand after surgery, which recovered after 3 months. One patient had poor healing of the surgical incision, which healed after symptomatic treatment. During the follow-up, the coronal and sagittal balance of all patients was maintained, and no other neurological complications occurred. There were no cases of screw loosening, broken screws, or broken rods, or other internal fixation failures.Conclusion:The application of asymmetric T 1 SPO technique in the treatment of stiff cervical thoracic lateral kyphosis deformity can achieve relatively satisfactory correction effects.

Objective:To compare the efficacy of whole-process visualization system-assisted pedicle screw internal fixation and free-hand pedicle screw internal fixation in the treatment of thoracolumbar burst fracture (TLBF) without neurologic symptoms.Methods:A retrospective cohort study was conducted to analyze the clinical data of 64 patients with TLBF without neurologic symptoms admitted to Zhengzhou Orthopedic Hospital from December 2020 to October 2022, including 41 males and 23 females, aged 23-52 years [(42.1±6.6)years]. The injured vertebrae involved T 11 in 26 patients, T 12 in 17, L 1 in 12, and L 2 in 9. The Wiltse approach was used in all the patients, 31 of whom were treated with pedicle screw internal fixation assisted by the whole-process visualization system (visualization system-assisted screw placement group) and 33 of whom were treated with free-hand pedicle screw internal fixation (free-hand screw placement group). The two groups were compared in terms of operation time, single screw placement time, intraoperative blood loss, intraoperative total radiation dose and total length of hospital stay. The accuracy of pedicle screw placement and penetration rate of the pedicle cortex were evaluated in the two groups. The Cobb angle and lumbar visual analogue scale (VAS) before surgery, at 1 week, 3 months after surgery and at the last follow-up were compared between the two groups. The incidence of postoperative complications was also investigated. Results:All the patients were followed up for 10-33 months [(17.5±4.8)months]. The operation time was (106.9±11.8)minutes in the visualization system-assisted screw placement group, shorter than (121.3±11.4)minutes in the free-hand screw placement group ( P<0.01). The single screw placement time was (9.1±1.0)minutes in the visualization system-assisted screw placement group, shorter than (11.7±1.5)minutes in the free-hand screw placement group ( P<0.01). The total radiation dose was (10.4±2.4)mGy in the visualization system-assisted screw placement group, lower than (51.8±7.2)mGy in the screw placement group ( P<0.01). There was no significant difference in intraoperative blood loss or total length of hospital stay between the two groups ( P>0.05). The accuracy of pedicle screw placement was 96.6% (197/204) in the visualization system-assisted screw placement group, significantly higher than 89.3% (191/214) in the free-hand screw placement group ( P<0.01). Both groups showed significant improvements in Cobb angle and VAS scores at 1 week, 3 months after surgery, and at the last follow-up ( P<0.05). There were no significant differences in Cobb angle or VAS scores between the two groups at each time point ( P>0.05). In the visualization system-assisted screw placement group, one patient had incision infection at 4 days after operation, which was cured with antibiotics. One patient in the free-hand screw placement group developed the symptoms of nerve root irritation at 2 days after surgery, which disappeared at 7 days after revision. There was no significant difference in the incidence of complications between the two groups ( P>0.05). During the follow-up, no patients had broken screws, loosening of internal fixation, or loss of correction in either group. Conclusions:Compared with free-hand pedicle screw internal fixation, the whole-process visualization system-assisted pedicle screw internal fixation in the treatment of TLBF without neurologic symptoms can shorten the time of operation and screw placement, reduce the radiation dose, and improve the accuracy of pedicle screw placement, suggesting that it is a safer and more effective auxiliary method for pedicle screw placement.

Objective:To investigate the feasibility and clinical efficacy of percutaneous vertebroplasty (PVP) with measured saturated bone cement injection in the treatment of elderly patients with stage Ⅱ Kümmell's disease.Methods:A retrospective analysis was conducted to analyze the clinical data of the 41 elderly patients with stage Ⅱ Kümmell's disease who had been treated at Department of Spinal Orthopedics, Zhengzhou Orthopedic Hospital from June 2017 to June 2023 by PVP with bone cement injection into the intravertebral vacuum cleft. According to the amount of bone cement injected, the patients were divided into a saturated volume group (bone cement injection metered ≥ 150% of the cleft volume preoperatively measured) in which there were 21 cases, 4 males and 17 females, aged (78.4±5.2) years and a conventional volume group (bone cement injection metered was 100% to 120% of the cleft volume preoperatively measured) in which there were 20 cases, 6 males and 14 females, aged (79.5±7.4) years. The operative time, vacuum cleft volume measured, actual volume of bone cement injected, and percentage of bone cement injected were compared between the 2 groups. Visual analogue scale (VAS) for pain and Oswestry disability index (ODI) were compared between preoperation, postoperative 3 days, and the final follow-up in the 2 groups, as well as between the 2 groups. Cement leakage and other complications were documented.Results:The differences in the preoperative general data were not statistically significant between the 2 groups, indicating comparability ( P>0.05). All the 41 elderly patients successfully completed their surgery. Follow-up time was (18.1±3.3) months. The operative time [(39.7±7.5) min], actual volume of bone cement injected [(5.6±0.9) mL], and percentage of bone cement injected (1.8%±0.3%) in the saturated volume group were all significantly greater than those in the conventional volume group [(35.5±4.9) min, (4.4±1.0) mL, and 1.2%±0.1%] ( P<0.05). Postoperatively, the incisions healed completely in all patients, with no such complications as cement-related adverse reactions. Cement leakage occurred in 2 patients in the conventional volume group, leading to lumbar pain or discomfort after activity, which was relieved by cement reinforcement and nail-rod internal fixation. VAS pain scores and ODIs at 3 d postoperatively and at the final follow-up were significantly improved in all patients compared with preoperation ( P<0.05). At the final follow-up, both VAS pain score and ODI in the saturated volume group improved significantly greater than those in the conventional volume group ( P<0.05). None of the patients had complications like cement displacement at the final follow-up. Conclusion:PVP with measured saturated bone cement injection into the intravertebral vacuum clefts is a safe and effective treatment for stage Ⅱ Kümmell's disease in elderly patients, offering a new minimally invasive option.

BACKGROUND:Previous studies found that the proliferative scar-specific long non-coding RNA lncRNA HSFAS is a novel biomarker that can be used in the diagnosis of hypertrophic scar,but how it functions in hypertrophic scar is not clear. OBJECTIVE:To investigate the role and mechanism of lncRNA HSFAS in hypertrophic scar.METHODS:Fresh scar tissue and surrounding normal skin tissue samples from three patients with hypertrophic scar were collected,and tissue immunofluorescence was used to detect the expression of lncRNA HSFAS in frozen sections of two skin tissues. Primary fibroblasts were isolated from proliferative scarred skin tissue and normal skin tissue and cultured by enzyme digestion method. Quantitative real-time PCR was used to detect the mRNA expression of lncRNA HSFAS in cells. The proteins bound to lncRNA HSFAS were detected by RNA pull-down combined mass spectrometry. GO and KEGG were used to analyze the main functions and pathways of lncRNA HSFAS involved in hypertrophic scar progression. The targeted binding of lncRNA HSFAS to proteins was determined by catRAPID and RPISeq website analysis. RESULTS AND CONCLUSION:Compared with normal skin tissue and fibroblasts from normal skin tissue,the expression of lncRNA HSFAS in human hypertrophic scar tissue and primary fibroblasts from hypertrophic scar tissue was significantly increased (P<0.05). There were 510 proteins clearly bound to lncRNA HSFAS by RNA pull-down combined mass spectrometry. The results of GO and KEGG analyses showed that these proteins were mainly involved in RNA splicing and processing,chromosome synthesis and separation,and cell cycle. Among them,the proteins involved in RNA splicing and processing included scaffold attachment factor B2 and DICER1,and the binding fraction with lncRNA HSFAS was higher. The results of bioinformatics analysis showed that lncRNA HSFAS was bound to scaffold attachment factor B2 and DICER1 proteins. To conclude,lncRNA HSFAS may affect gene expression by interacting with scaffold attachment factor B2 and DICER1 proteins to regulate RNA splicing and processing modification,thus promoting the occurrence and development of hypertrophic scar.