ObjectiveTo investigate the effect and potential mechanism of caffeic acid （CA） on severe acute pancreatitis （SAP） induced by caerulein combined with lipopolysaccharide （LPS）， and to provide a basis for the research on novel drugs for the treatment of SAP. MethodsC57BL/6J mice， aged 6 weeks， were divided into control group， model group， CA group， and octreotide acetate （OA） group， with 6 mice in each group. The mice in the control group were given injection of normal saline， and those in the other groups were given intraperitoneal injection of caerulein combined with LPS to establish a mouse model of SAP. At 1 hour after the first injection of caerulein， the mice in the CA group and the OA group were given intraperitoneal injection of CA or subcutaneous injection of OA at an interval of 8 hours. The general status of the mice was observed after 24 hours of modeling， and serum， pancreas， lung， and colon samples were collected. HE staining was used to observe the histopathological changes of the pancreas and lungs， and the serum levels of α-amylase， lipase， tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， alanine aminotransferase， aspartate aminotransferase， and creatinine were measured. RT-PCR was used to measure the expression of proinflammatory factors in the pancreas and lungs； myeloperoxidase （MPO） immunohistochemistry was used to observe the degree of neutrophil infiltration； Western blot was used to measure the activation of nuclear factor-kappa B （NF-κB） and the level of citrullinated histone H3 （CitH3）， a marker for the formation of neutrophil extracellular traps （NETs）， in the pancreas and lungs， as well as the expression level of ZO-1 in colon tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the Dunnett’s t-test was used for further comparison between two groups. ResultsCompared with the control group， the model group had severe injury in the pancreas and lungs and significant increases in the activity of serum α- amylase and lipase and the levels of the proinflammatory cytokines IL-6， interleukin-1β （IL-1β）， and TNF-α in serum and lung tissue （all P<0.05）， as well as significant increases in NF-κB activation， neutrophil infiltration， and the formation of NETs in the pancreas and lungs （all P<0.05）. Compared with the model group， the CA group had alleviated pathological injury of the pancreas and lungs and significant reductions in the activity of serum α-amylase and the levels of the proinflammatory cytokines IL-6， IL-1β， and TNF-α in serum and lung tissue （all P<0.05）， as well as significant reductions in NF-κB activation， neutrophil infiltration， and the formation of NETs in the pancreas and lungs （all P<0.05）. ConclusionCA can alleviate SAP induced by caerulein combined with LPS in mice， possibly by inhibiting neutrophil recruitment and the formation of NETs.

OBJECTIVE: Benzylisoquinoline alkaloids (BIAs) have pharmacological functions and clinical use. BIAs are mainly distributed in plant species across the order Ranunculales and the genus Phellodendron from Sapindales. The BIA biosynthesis has been intensively investigated in Ranunculales species. However, the accumulation mechanism of BIAs in Phellodendron is largely unknown. The aim of this study is to unravel the biosynthetic pathways of BIAs in Phellodendron amurens. METHODS: The transcriptome and metabolome data from 18 different tissues of P. amurense were meticulously sequenced and subsequently subjected to a thorough analysis. Weighted gene co-expression network analysis (WGCNA), a powerful systems biology approach that facilitates the construction and subsequent analysis of co-expression networks, was utilized to identify candidate genes involved in BIAs biosynthesis. Following this, recombinant plasmids containing candidate genes were expressed in Escherichia coli, a widely used prokaryotic expression system. The purpose of this genetic engineering endeavor was to express the candidate genes within the bacteria, thereby enabling the assessment of the resultant enzyme activity. RESULTS: The synonymous substitutions per synonymous site for paralogs indicated that at least one whole genome duplication event has occurred. The potential BIA biosynthetic pathway of P. amurense was proposed, and two PR10/Bet v1 members, 14 CYP450s, and 33 methyltransferases were selected as related to BIA biosynthesis. One PR10/Bet v1 was identified as norcoclaurine synthase, which could catalyze dopamine and 4-hydroxyphenylacetaldehyde into (S)-norcoclaurine. CONCLUSION Our studies provide important insights into the biosynthesis and evolution of BIAs in non-Ranunculales species.

Traditional Chinese medicine (TCM) serves as a treasure trove of ancient knowledge, holding a crucial position in the medical field. However, the exploration of TCM's extensive information has been hindered by challenges related to data standardization, completeness, and accuracy, primarily due to the decentralized distribution of TCM resources. To address these issues, we developed a platform for TCM knowledge discovery (TCMKD, https://cbcb.cdutcm.edu.cn/TCMKD/). Seven types of data, including syndromes, formulas, Chinese patent drugs (CPDs), Chinese medicinal materials (CMMs), ingredients, targets, and diseases, were manually proofread and consolidated within TCMKD. To strengthen the integration of TCM with modern medicine, TCMKD employs analytical methods such as TCM data mining, enrichment analysis, and network localization and separation. These tools help elucidate the molecular-level commonalities between TCM and contemporary scientific insights. In addition to its analytical capabilities, a quick question and answer (Q&A) system is also embedded within TCMKD to query the database efficiently, thereby improving the interactivity of the platform. The platform also provides a TCM text annotation tool, offering a simple and efficient method for TCM text mining. Overall, TCMKD not only has the potential to become a pivotal repository for TCM, delving into the pharmacological foundations of TCM treatments, but its flexible embedded tools and algorithms can also be applied to the study of other traditional medical systems, extending beyond just TCM.

OBJECTIVE: Anemoside B4 (AB4), the most abundant triterpenoidal saponin isolated from Pulsatilla chinensis, inhibited influenza virus FM1 or Klebsiella pneumoniae-induced pneumonia. However, the anti-SARS-CoV-2 effect of AB4 has not been unraveled. Therefore, this study aimed to determine the antiviral activity and potential mechanism of AB4 in inhibiting human coronavirus SARS-CoV-2 in vivo and in vitro. METHODS: The cytotoxicity of AB4 was evaluated using the Cell Counting Kit-8 (CCK8) assay. SARS-CoV-2 infected HEK293T, HPAEpiC, and Vero E6 cells were used for in vitro assays. The antiviral effect of AB4 in vivo was evaluated by SARS-CoV-2-infected hACE2-IRES-luc transgenic mouse model. Furthermore, label-free quantitative proteomics and bioinformatic analysis were performed to explore the potential antiviral mechanism of action of AB4. Type I IFN signaling-associated proteins were assessed using Western blotting or immumohistochemical staining. RESULTS: The data showed that AB4 reduced the propagation of SARS-CoV-2 along with the decreased Nucleocapsid protein (N), Spike protein (S), and 3C-like protease (3CLpro) in HEK293T cells. In vivo antiviral activity data revealed that AB4 inhibited viral replication and relieved pneumonia in a SARS-CoV-2 infected mouse model. We further disclosed that the antiviral activity of AB4 was associated with the enhanced interferon (IFN)-β response via the activation of retinoic acid-inducible gene I (RIG-1) like receptor (RLP) pathways. Additionally, label-free quantitative proteomic analyses discovered that 17 proteins were significantly altered by AB4 in the SARS-CoV-2 coronavirus infections cells. These proteins mainly clustered in RNA metabolism. CONCLUSION Our results indicated that AB4 inhibited SARS-CoV-2 replication through the RLR pathways and moderated the RNA metabolism, suggesting that it would be a potential lead compound for the development of anti-SARS-CoV-2 drugs.

Objective:To investigate the clinical significance of intraoperative prepositioning of dual mediastinal drains in elderly patients developing anastomotic leakage(AL)after a total endoscopic Ivor-Lewis procedure.Methods:This retrospective case-control study analyzed the clinical data of 500 elderly patients who underwent total endoscopic Ivor-Lewis surgery for esophageal or cardia cancer from January 2020 to December 2022.In the control group, one mediastinal drainage tube was placed intraoperatively, while in the study group, two mediastinal drainage tubes were placed.Both groups had a chest tube placed conventionally.The study compared the incidence of anastomotic leak(AL)at 1 month postoperatively, inflammatory indexes in patients with AL, grading of AL, rate of nasal fistula placement, incision infection, anastomotic stenosis, and incidence of hoarseness.Additionally, it compared ICU occupancy, ventilator use, and ICU length of stay between the two groups.Results:The analysis included clinical data from 455 elderly patients.Among the patients who developed AL, the study group had significantly lower peak body temperature[(39.58±1.03)℃ vs.(38.05±0.56)℃, t=4.298, P<0.05], white blood cell count[(18.63±3.35)×10 9/L vs.(14.28±2.78)×10 9/L, t=3.450, P<0.05], and C-reactive protein(CRP)levels[(154.66±41.64)mg/L vs.(122.19±31.29)mg/L, t=2.131, P<0.05]. The study group also had a significantly lower grading of AL and rate of nasal fistula placement(82.4% vs.30.0%, P<0.05). In terms of ICU indicators, the study group had a significantly lower ICU admission rate(64.7% vs.10.0%, P<0.05), shorter period of ventilator use[(6.47±8.15)days vs.(0.90±2.23)days, t=2.62, P<0.05], and shorter ICU stay[(11.70±8.89)days vs.(4.70±6.27)days, t=2.184, P<0.05]. Conclusions:Double mediastinal drainage tubes, have been found to significantly alleviate inflammation, decrease the rate of nasal fistula placement and ICU admission, and shorten the length of ICU stay in elderly patients.Therefore, they are considered safe and deserving of clinical promotion.

Objective To investigate the effect and mechanism of cinobufotalin in inhibiting hepatocellular carcinoma(HCC)metastasis by regulating epithelial-mesenchymal transition(EMT).Methods A total of 36 male BALB/c nude mice,aged 6 weeks,were given injection of MHCC97H cells via the caudal vein to establish a model of HCC lung metastasis,and then the mice were randomly divided into high-and low-dose cinobufotalin groups and control group.Since the day of modeling,the mice in the high-and low-dose cinobufotalin groups were given intraperitoneal injection of cinobufotalin at a dose of 120 μL/kg and 60 μL/kg,respectively,and those in the control group were given intraperitoneal injection of normal saline,twice a week.After 8 weeks,HE staining was performed for lung tissue to measure the lung metastasis rate of HCC.MHCC97H cells were treated with high-dose(2.5 μL/mL)or low-dose(5 μL/mL)cinobufotalin for 24 hours,and wound healing assay,RT-PCR,and Western blot were used to measure cell migration ability and the expression of EMT-related molecules.MHCC97H cells were induced in a simulated hypoxic environment with CoCl2 incubation,with high-and low-dose cinobufotalin added for intervention,and wound healing assay and Western blot were used to investigate the effect of cinobufotalin on cell migration ability and EMT induced by hypoxia.Transcriptome analysis was used to investigate the effect mechanism of cinobufotalin on MHCC97H cells,and Western blot was used to observe the effect of cinobufotalin on the expression levels of protein kinase B(AKT)and phosphorylated AKT(P-AKT)in MHCC97H cells.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups;the independent-samples t test was used for comparison of categorical data between two groups.Results Compared with the control group,the cinobufotalin group had a significant reduction in the lung metastasis rate of HCC.Compared with the control group,cinobufotalin intervention reduced the wound healing rate of MHCC97H cells,upregulated the expression of epithelial-type molecules(t=2.860,P<0.05),and downregulated the expression of EMT transcription factors(EMT-TFs)and mesenchymal molecules(t=3.545,2.022,2.852,and 2.341,all P<0.05).Hypoxia induction upregulated the wound healing rate of MHCC97H cells and the expression levels of mesenchymal molecules and EMT-TFs(P<0.05),and cinobufotalin intervention reversed EMT change and inhibited wound healing(P<0.05).The transcriptome analysis of MHCC97H cells showed significant gene differences between the cinobufotalin group and the control group,and cinobufotalin mainly affected the expression of genes associated with tumor,metabolism,immunity,and signal transduction,with the largest number of differentially expressed genes in the AKT signal transduction pathway.Further measurement showed that cinobufotalin intervention downregulated the expression levels of AKT,P-AKT,and P-AKT/AKT in MHCC97H cells(t=2.434,3.401,and 2.258,all P<0.05).Conclusion Cinobufotalin can inhibit the metastasis of HCC,especially hypoxia-induced HCC metastasis,and regulation of EMT mediated by the AKT signal transduction pathway in HCC cells might be one of its mechanisms of action.

The training of orthopedic residents in Taiwan, China includes post graduate year and specialist training, which contain continuous orthopedic specialist training, humanistic training, and holistic education to ensure the high post competency of orthopedic residents. The training process is oriented towards competency and outcome, emphasizing the cultivation of core competencies. Comprehensive quantitative standards have been established to comprehensively evaluate the post competency of residents through theoretical, technical, and clinical work assessments. In this paper, the literature on medical education and resident training in Taiwan was reviewed. The overall training system, assessment requirements, and humanistic training of orthopedic residents in Taiwan were introduced. In addition, the models of training orthopedic specialists were compared between Taiwan and Mainland of China represented by Peking University Medical Department to provide reference for the training program of orthopedic residents.

Objective To explore the genetic polymorphism and mutation rate of 36 Y-STR loci in Sichuan Han population and provide basic data for their forensic applications.Methods Blood samples were collected from 307 unrelated male individuals and 367 father-son pairs in Sichuan Han population using FTA blood sampling cards.The Y41SE-v1.2 kit was used to type 36 Y-STR loci,and the population genetic parameters and mutation rates of each locus were calculated.The genetic relationship between Sichuan Han population and other ethnic populations was analyzed in combination with 19 other populations in China.Results A total of 79 mutations were observed in 36 Y-STR loci in 367 father-son pairs of Sichuan Han population,with an average mutation rate of 6.00×10-3.There were 74(93.67%)one-step mutations and 5(6.33%)multi-step mutations.A total of 383 alleles were found at 36 Y-STR loci in 307 Sichuan Han males.The number of alleles at each locus ranged from 5(0.016 29)to 56(0.182 41),GD ranged from 0.100 02(DYS645)to 0.959 99(DYS385),and 306 haplotypes were found.The HD,DC and HMP values were 0.999 973,99.67%and 0.003 284,respectively.The results of genetic distance(Fst)analysis showed that the Sichuan Han population had the closest genetic distance with the Chongqing Han population(0.000 9)and the furthest genetic distance with the Xinjiang Mongolian group(0.032 0).The MDS based on genetic distance was basically consistent with the results of cluster analysis.Conclusion The 36 Y-STR loci have high genetic polymorphism in the Sichuan Han population.The study data can provide data support for the construction of the Y-STR database in this region,and also provide basic genetic information for forensic investigation and population genetics research.

Objective:The Nomogram prediction model of secondary insomnia in adolescent depression patients was constructed to provide reference and basis for clinical prevention of secondary insomnia in adolescent depression patients.Methods:A total of 158 adolescent depression patients who were treated in the Department of Pediatric Psychiatry of Shaoxing Seventh People′s Hospital from May 2021 to March 2022 were selected as the research objects by convenient sampling method. According to the Pittsburgh Sleep Quality Index (PSQI) score, the group was divided into a normal sleep group (PSQI ≤ 7 points) and a secondary insomnia group (PSQI>7 points). A cross-sectional survey was conducted using the Hamilton Depression Scale, Hamilton Anxiety Scaleand Social Support Rating Scale. Multivariate binary Logistic regression was used to analyze the influencing factors of secondary insomnia and construct a Nomogram prediction model for secondary insomnia in adolescents with depression. The accuracy of the model was judged by receiver operating characteristic curve. In addition, 70 adolescent depression patients who were treated in Shaoxing Seventh People′s Hospital from May 2022 to March 2023 were selected for success rate prediction. The results were compared with the actual observed results, and the fitting degree of the model was evaluated by Hosmer-Lemeshow test.Results:There were 72 cases in the normal sleep group, including 40 males and 32 females, aged (15.34 ± 2.62) years old, and 86 cases in the secondary insomnia group, including 51 males and 35 females, aged (14.89 ± 2.13) years old. The results of multivariate analysis showed that depression ( OR=6.381, 95 % CI 1.548-26.295, P<0.05), anxiety ( OR=6.248, 95 % CI 1.445-27.005, P<0.05) and social support ( OR=0.586, 95 % CI 0.346-0.994, P<0.05) were independent influencing factors for the occurrence of secondary insomnia in the two groups. On this basis, a risk prediction model was established. The AUC was 0.999, the sensitivity was 98.80 %, the specificity was 98.60 %, and the Youden index was 0.974. The total prediction accuracy of the external validation model was 99.80 %. Conclusions:The Nomogram prediction model of secondary insomnia in adolescent depression patients has excellent predictive efficacy. The degree of depression, anxiety and social support are the main influencing factors of secondary insomnia in patients. Medical staff should provide more effective psychological support for patients.