The culture of suspended Vero cells is facing difficulties such as low cell viability and long doubling time. To investigate the main reasons for the slow growth and low viability of suspended Vero cells, this study conducted transcriptomic analysis of suspended Vero cells (Vero-XF) and adherent Vero cells (Vero-AD) to screen the differentially expressed genes (DEGs) affecting the growth of suspended cells. In addition, epidermal growth factor (EGF) was supplemented to the culture system to improve the growth of Vero-XF. The results showed that compared with the Vero-AD group, the Vero-XF group had 7 376 significant DEGs. Kyoto encyclopedia of genes and genomes enrichment analysis revealed that the DEGs were mainly enriched in the autophagy and mitophagy pathways. Eleven DEGs were selected and verified by quantitative real-time PCR, which showed up-regulated expression of ATG9B, WIPI2, LAMP2, OPTN, Rab7a, and DEPTOR and down-regulated expression of ATG4D, being consistent with the results of transcriptomic analysis. In addition, the Vero-XF group showed significantly up-regulated expression of ATG101, ATG2A, and STX17 and insignificant change in the expression of NBR1, compared with the Vero-AD group. The protein levels of LC3 and P62 in Vero-XF and Vero-AD were determined by Western blotting, which showed up-regulated expression of LC3Ⅱ/Ⅰ and down-regulated expression of P62 in Vero-XF, indicating a higher level of autophagy. Finally, the exogenous supplementation of EGF at 10, 20, and 30 μg/L in the culture system reduced the autophagy level of Vero-XF by 22.35%, 48.15%, and 71.29%, increased the specific growth rate by 15.48%, 33.33%, and 57.14%, and decreased the apoptosis rate by 2.84%, 15.46%, and 16.23%, respectively. The results of this study preliminarily reveal that the activation of autophagy is one of the reasons for the slow growth of Vero-XF, which provides reference for the subsequent culture of suspended Vero cells.
Animals ; Vero Cells ; Autophagy/genetics* ; Chlorocebus aethiops ; Epidermal Growth Factor/pharmacology* ; Gene Expression Profiling ; Transcriptome ; Cell Survival

In recent years, significant progress has been made in pharmacological research on the treatment of ischemic stroke with monomeric components of traditional Chinese medicine, among which flavonoids have shown good neuroprotective effects. This article reviews the regulatory role of flavonoids in the signaling pathway of ischemic brain injury.

Objective:To study the clinicopathological features of Sjogren′s syndrome (SS) with liver injury and to improve the understanding of this disease.Methods:Forty-nine patients with SS complicated with liver injury were collected from Beijing Ditan Hospital, Capital Medical University from October 2008 to January 2022. All patients underwent ultrasound-guided liver biopsy, and all specimens were stained with HE. The histopathologic characteristics were observed and the pathologic indexes were graded. Immunohistochemical stains for CK7, CK19, CD38, MUM1 and CD10 were performed by EnVision method; and special histochemical stains for reticulin, Masson′s trichrome, Rhodanine, Prussian blue, periodic acid Schiff (PAS) and D-PAS stains were conducted .Results:The age of patients ranged from 31 to 66 years, including 3 males and 46 females. SS combined with drug-induced liver injury was the most common (22 cases, 44.9%), followed by autoimmune liver disease (13 cases, 26.5%, including primary biliary cholangitis in eight cases, autoimmune hepatitis in 3 cases, and PBC-AIH overlap syndrome in 2 cases), non-alcoholic fatty liver disease (NAFLD, 9 cases, 18.4%) and other lesions (5 cases, 10.2%; including 3 cases of nonspecific liver inflammation, 1 case of liver amyloidosis, and 1 case of porto-sinusoidal vascular disease). Among them, 28 cases (57.1%) were associated with obvious interlobular bile duct injury, mainly in SS combined with PBC group and drug-induced liver injury group. Twenty-three cases (46.9%) were associated with hepatocyte steatosis of varying degrees. In SS with autoimmune liver disease group, ISHAK score, degree of fibrosis bile duct injury, bile duct remodeling, lymphocyte infiltration of portal area, and plasma cell infiltration, MUM1 and CD38 expression; serum ALP and GGT, IgM; elevated globulin; positive AMA, proportion of AMA-M2 positive and IgM positive were all significantly higher than those in other groups(all P<0.05). Serum ALT, direct bilirubin and SSA positive ratio in SS combined with drug liver group were significantly higher than those in other groups(all P<0.05). The serum total cholesterol level in SS combined with PBC group ( P=0.006) and NALFD group ( P=0.011) were significantly higher than those in other groups ( P<0.05). Conclusions:The pathologic manifestations of SS patients with liver injury are varied. The inflammatory lesions of SS patients with autoimmune liver disease are the most serious, and the inflammatory lesions of SS patients with non-alcoholic fatty liver disease and non-specific inflammation are mild. Comprehensive analysis of liver histopathologic changes and laboratory findings is helpful for the diagnosis of SS complicated with different types of liver injury.

Objective:To investigate the clinicopathological characteristics of rashes in monkeypox patients through a series of skin biopsies, and examine their pathological features and the most effective tests.Methods:Patients with monkeypox virus infection admitted to Beijing Ditan Hospital from June to August 2023 were identified. Among them, 24 patients underwent skin biopsies for clinical pathological study that were included in this study. Clinical information, rash pictures, and nucleic acid test results were analyzed using histopathology, immunohistochemistry, RNAscope ? hybridization and electron microscopy. Results:All 24 patients were male, including 14 patients with concurrent human immunodeficiency virus infection. Their average age was (32.3±5.4) years. The nucleic acid test confirmed monkeypox virus infection. The clinical feature of monkeypox rashes was solitary rather than clustered distribution, with rashes occurring in similar phase, distinguishing it from herpesvirus. The rashes in these patients were mostly scattered, with an average of (13.0±11.8) rashes, and most commonly present in the perineum, face, limbs, and trunk. The three main pathological features of these rashes were ballooning degeneration of the epidermal spinous cell layer, the characteristic intra-cytoplasmic Guarnieri′s bodies and significant infiltration of inflammatory cells in whole dermal layer. Immunohistochemistry, RNAscope ? hybridization, and electron microscopy can all effectively detect the monkeypox virus. Electron microscopy showed viral replication in various types of skin cells. Conclusions:The study describes the pathological features of monkeypox virus rashes. Pathological examination of skin biopsy samples is helpful to diagnose these rashes. The study suggests that the monkeypox virus has a unique epitheliotropic affinity and can infect various types of cells in the skin.

Objective:To examine the burden and trends of acute viral hepatitis in Guangdong Province from 1990 to 2019, and provide reference evidences for hepatitis prevention and control in the province.Methods:Data on acute viral hepatitis (hepatitis A, B, C, and E) in Guangdong from 1990 to 2019 were extracted from the Global Burden of Disease Study 2019 database. The incidence, prevalence, mortality, and disability-adjusted life years (DALY) data were analyzed by age and gender, and the estimated annual percentage change (EAPC) was calculated to describe the changing trends in disease burden.Results:From 1999 to 2019, the standardized incidence, prevalence, mortality, and DALY of acute viral hepatitis in Guangdong were higher than the national averages. In 2019, 51.43% (2 245 087/4 365 221) of acute viral hepatitis cases in Guangdong Province were mainly attributed to hepatitis B, and 77.18% (106/138) of deaths were due to acute hepatitis B. In different age groups, except for acute hepatitis B, which was more common in adults, the incidence rates of other types of viral hepatitis such as hepatitis A, B, and E showed an overall decreasing trend with age. The mortality rates of different types of acute viral hepatitis, except for the <5 age group, increased with age. The overall incidence and mortality rates of acute viral hepatitis were higher in men than in women.Conclusions:The overall burden of acute viral hepatitis in Guangdong declined in 2019, but remained higher than the national level. Further efforts are needed to strengthen hepatitis prevention and screening in different population in Guangdong Province, especially in children and the elderly.

Hepatocellular carcinoma(HCC)and biliary tract cancer(BTC)are significant health challenges in China because of their high prevalence and mortality rates.Advances in molecular diagnostics have opened new avenues for personalized treatment strategies.This consensus provides a comprehensive update on the clinical applications of molecular diagnostics in HCC and BTC and addresses the urgent need for personalized treatment strategies tailored to the Chinese population,emphasizing the importance of molecular markers in guiding targeted therapies and immunotherapies.By employing the Delphi method and the Grading of Recommendations Assessment,Development,and Evaluation system,the expert panel formulated evidence-based recommendations for the use of molecular diagnostics in the clinical management of HCC and BTC.Key molecular markers,such as isocitrate dehydrogenase(IDH)1 and 2,fibroblast growth factor receptor 2(FGFR2),BRAF V600E,human epidermal growth factor receptor 2(HER2),rearranged during transfection(RET),and neurotrophic tyrosine receptor kinase(NTRK),are highlighted for their roles in optimizing treatment regimens.The consensus also explores the signifi-cance of emerging biomarkers,such as tumor mutation burden and microsatellite instability,in pre-dicting responses to immune checkpoint inhibitors.The recommendations aim to enhance precision medicine approaches,improve patient outcomes,and foster the integration of molecular diagnostics into routine clinical practice.

Objective To investigate the predictive value of preoperative fibrinogen/albumin ratio (FAR) and systemic immune inflammation index (SII) on the postoperative prognosis of patients with pancreatic ductal adenocarcinoma. Methods An ROC curve was used in determining the best cutoff values of FAR and SII and then grouped. The Cox proportional hazards model was used in analyzing the prognostic factors of radical pancreatic cancer surgery, and then a Nomogram prognostic model was established. C-index, AUC, and calibration curve were used in evaluating the discrimination and calibration ability of the Nomogram. DCA curves were used in assessing the clinical validity of the Nomograms. Results The optimal cutoff values for preoperative FAR and SII were 0.095 and 532.945, respectively. FAR≥ 0.095, SII≥ 532.945, CA199≥ 450.9 U/ml, maximum tumor diameter≥ 4 cm, and the absence of postoperative chemotherapy were independent risk factors for the poor prognosis of pancreatic cancer (P<0.05). The discrimination ability, calibration ability, and clinical effectiveness of Nomogram prognostic model were better than those of the TNM staging system. Conclusion The constructed Nomogram prognostic model has higher accuracy and level of discrimination and more clinical benefits than the TNM staging prognostic model.

OBJECTIVE: To explore the mechanisms that mediate the anti-inflammatory activity of Eurycoma longifolia. METHODS: Kunming mouse models of xylene-induced ear swelling and lipopolysaccharide (LPS)-induced acute pneumonia were used to compare the anti- inflammatory activities of aqueous and ethanol extracts of Eurycoma longifolia. UPLC-Q-TOF-MS/MS was used to identify the chemical composition in the ethanol extract of Eurycoma longifolia, based on which the potential antiinflammatory targets of Eurycoma longifolia were screened using the databases including SwissADME, SwissTargetPrediction, and Genecards. The String database was used to generate the protein-protein interaction (PPI) network, and Cytoscape was used for network topology analysis and screening the core targets. The enrichment of the core targets was analyzed using Metascape database, the core components and targets were docked with Autodock software, and the docking results were visualized using Pymol software. In a RAW264.7 cell model of LPS-induced inflammation, the Griess reagent was used to measure NO level, and Western blotting was performed to detect the expression levels of MAPK1, JAK2, and STAT3 proteins to verify the anti- inflammatory mechanism of Eurycoma longifolia. RESULTS: The ethanol extract (75%) of Eurycoma longifolia (ELE) was the active site, which contained a total of 37 chemical components. These chemical compounds and diseases had 541 targets, involving the JAK/STAT3, cAMP and other signaling pathways. Twelve indicator components were identified, which all showed good results of molecular docking with two core targets involved in the signaling pathways. In the cell validation experiment, treatment of the cells with low-, medium-, and high-dose ELE significantly reduced NO release in the cells, and ELE at the medium dose significantly decreased the cellular expressions of JAK2 and STAT3. CONCLUSION The anti-inflammatory activity of Eurycoma longifolia is attributed primarily to its active ingredients bitter lignin and alkaloids, which may regulate the JAK/STAT3 signaling pathway by targeting JAK2 and STAT3.
Animals ; Mice ; Network Pharmacology ; Eurycoma ; Lipopolysaccharides ; Molecular Docking Simulation ; Tandem Mass Spectrometry ; Anti-Inflammatory Agents/pharmacology* ; Ethanol ; Plant Extracts/pharmacology*

Objective:To investigate the safety and efficacy of ultrasound-guided sclerotherapy combined with radiofrequency ablation on the complex lymphatic malformations (LM) in children.Methods:The clinical data of 21 children with complex LM treated with ultrasound-guided sclerotherapy combined with radiofrequency ablation in the First Affiliated Hospital of Zhengzhou University from June 2018 to October 2021 were retrospectively analyzed.Intraoperative and postoperative complications were recorded.Imaging examinations were performed at 1, 3, 6, 9, 12, 18, 24 months postoperatively to observe the recurrence, the volume of the lesions and their reduction rate were calculated, and the efficacy was analyzed. Friedman test was used to compare the lesion volume at different time points before and after surgery, and the reduction rate of lesion volume at 1 month postoperatively and other time points after surgery. Results:A total of 21 children were included in this study, among them, there were 12 males and 9 females, age range from 1 month to 5 years and 6 months, with a median age of 23 months.A total of 26 LM in 21 children were successfully treated, and no serious complications like organ damage occurred during and after surgery.One patient with abdominal LM had a postoperative infection, which was controlled by 3 weeks of catheter drainage.Four LM in 3 children recurred at 3 or 6 months after surgery, while all lesions were significantly narrowed down than those before surgery and they were cured after 1-3 sessions of continued sclerotherapy.There were significant differences in the lesion volumes before surgery and 1, 3, 6, 9, 12, 18 and 24 months postoperatively [222.26(159.57, 316.40) cm 3vs.43.06(22.74, 62.53) cm 3, 31.56(15.49, 45.94) cm 3, 25.21(9.63, 36.22) cm 3, 19.80(6.79, 28.81) cm 3, 12.80(3.93, 20.38) cm 3, 7.13(0, 11.34) cm 3, and 2.79(0, 4.93) cm 3; all P<0.05]. There were significant differences between the volume reduction rates at 1 month postoperatively and 3, 6, 9, 12, 18, and 24 months postoperatively [79.36(73.30, 87.81)% vs.85.40(81.09, 91.61)%, 88.85(84.70, 93.61)%, 91.67(87.87, 95.05)%, 94.15(94.47, 97.35)%, 97.11(95.02, 100.00)%, and 99.04(97.93, 100.00)%; all P<0.05]. Patients were followed up for 24 months, and all of them were cured. Conclusions:Ultrasound-guided sclerotherapy combined with radiofrequency ablation is a minimally invasive, safe and effective therapeutic strategy for children with complex LM.

Objective:To investigate the predictive value of preoperative lymphocyte-to-monocyte ratio (LMR) combined with platelet-to-lymphocyte ratio (PLR) (LMR-PLR) scoring model for prognosis of pancreatic ductal adenocarcinoma (PDAC) after radical resection.Methods:The retrospective cohort study was conducted. The clinicopathological data of 116 patients with PDAC who were admitted to the Second Hospital of Lanzhou University from January 2015 to December 2019 were collected. There were 73 males and 43 females, aged 61.5(range, 29.0-75.0)years. All patients underwent radical resection for PDAC. Observation indicators: (1) optimal cut-off value of LMR and PLR; (2) clinicopathological features of patients with different scores of preoperative LMR-PLR scoring model; (3) follow-up and survival; (4) influencing factors for prognosis of PDAC patients; (5) construction and verification of nomogram prediction model. Measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test. Comparison of ordinal data was conducted using the Mann-Whitney U test. The Graphpad prism 8 was used to draw survival curve, the Kaplan-Meier method was used to calculate survival rate, and the Log-Rank test was used for survival analysis. The COX proportional hazard regression model was used for univariate and multivariate analyses. The X-tile software was used to determine the optimal cut-off values of LMR and PLR. The nomogram prediction model was conducted based on the results of multivariate analysis, and the receiver operating characteristic (ROC) curve was drawn. The area under curve (AUC) was used to evaluate the discrimination of nomogram prediction model. The calibration curve was used to evaluate the consistency of nomogram prediction model and the decision curve was used to evaluate the clinical benefits. Results:(1) Optimal cut-off value of LMR and PLR. The optimal cut-off values of LMR and PLR were 1.9 and 156.3. (2) Clinicopathological features of patients with different scores of preoperative LMR-PLR scoring model. Cases with LMR-PLR scoring as 0, 1, 2 were 11, 42, 63. Cases with CA125 <12.4 U/mL, cases postoperative with vascular invasion, cases with postoperative chemotherapy in patients with 0, 1, 2 of LMR-PLR scoring were 1, 8, 24, 9, 27, 27, 3, 26, 43, showing significant differences among them ( χ2=6.73, 8.37, 6.68, P<0.05). (3) Follow-up and survival. All 116 patients were followed up for 39(range, 2-86)months. The 1-, 2-, 3-year survival rate of 116 PDAC patients was 50.9%, 37.9%, 19.3%, respectively, with a survival time of 13(range, 1-85)months. The survival time of patients with LMR-PLR scoring as 0, 1, 2 was 3(range, 1-9)months, 7(range, 2-56)months, 26(range, 2-85)months, respectively, showing a significant difference among them ( χ2=48.78, P<0.05). (4) Influencing factors for prognosis of PDAC patients. Results of multivariate analysis showed that carcinoembryonic antigen (CEA), CA19-9, LMR-PLR score, tumor diameter were independent factors affecting prognosis of patients ( hazard ratio=1.61, 1.88, 0.27, 1.87, 95% confidence interval as 1.02-2.54, 1.18-3.00, 0.19-0.39, 1.13-3.09, P<0.05). (5) Construction and verification of nomogram prediction model. The nomogram prediction model was constructed based on CEA, CA19-9, LMR-PLR score and tumor diameter. The AUC of ROC curve in predicting 1-, 2-, 3-year survival rate of patients was 0.86 (95% confidence interval as 0.79-0.93, P<0.05), 0.86 (95% confidence interval as 0.79-0.92, P<0.05), 0.87 (95% confidence interval as 0.78-0.95, P<0.05), respectively. Results of calibration curve showed that the predicted survival rate of nomogram prediction model was consistent with the actual survival rate, with the consistency index as 0.74. Results of decision curve showed that the predictive performance of nomogram prediction model was superior to that of a single factor at a risk threshold of 0.12-0.85. Conclusions:CEA, CA19-9, LMR-PLR score, tumor diameter are independent factors affecting prognosis of patients undergoing radical resection for PDAC, and the nomogram prediction model can predict postoperative survival rate. The predicted survival rate of nomogram prediction model is consistent with the actual survival rate, and the predictive performance of nomogram prediction model is superior to that of a single factor at a risk threshold of 0.12-0.85.