ObjectiveTo evaluate the antitumor activity of Periplaneta americana extract（PAE） against human-derived lung adenocarcinoma organoids（LUAD-PDOs） and to elucidate its potential mechanism based on transcriptomics. MethodsFresh tumor and adjacent normal tissues from patients with LUAD were collected to construct LUAD-PDOs and normal lung organoid（Nor-PDOs） models using 3D organoid culture technology. The effective intervention concentration of PAE was determined using the cell counting kit-8（CCK-8） assay. Experimental groups included the model group（LUAD-PDOs）， normal group， model administration group（LUAD-PDOs+PAE）， and normal administration group（Nor-PDOs+PAE）. Hematoxylin-eosin（HE） staining was used to observe the pathological structures of PDOs， immunohistochemistry（IHC） was performed to detect the expressions of the proliferation marker Ki-67 and lung adenocarcinoma differentiation markers cytokeratin-7（CK-7） and Napsin A， TUNEL staining was applied to detect cell apoptosis. RNA sequencing（RNA-Seq） was conducted to identify differentially expressed genes（DEGs）， followed by Gene Ontology（GO）， Kyoto Encyclopedia of Genes and Genomes（KEGG）， and Gene Set Enrichment Analysis（GSEA）， alongside protein-protein interaction（PPI） network analysis to screen core mechanisms. Finally， key targets were validated by integrating external database analysis with immunofluorescence（IF）. ResultsNor-PDOs and LUAD-PDOs that highly recapitulated the pathological characteristics of the primary tissues were successfully established. The CCK-8 assay determined that the effective intervention concentration of PAE was 16 g·L^-1. Morphological observation showed that Nor-PDOs exhibited lumen-forming structures， whereas LUAD-PDOs displayed dense， solid structures. CCK-8 and TUNEL assays revealed that， compared with the model group， PAE intervention inhibited the proliferation of LUAD-PDOs and promoted apoptosis in LUAD cells， while showing no significant effect on the viability of Nor-PDOs. Transcriptomic analysis identified 719 DEGs that were significantly reversed after PAE intervention（347 up-regulated and 372 down-regulated）（P<0.05）. GO enrichment analysis indicated that DEGs in the model administration group were significantly enriched in biological processes related to cell cycle regulation compared to the model group. KEGG pathway analysis revealed that PAE affected pathways related to proliferation and metabolism， including pathways in cancer and the p53 signaling pathway. GSEA further confirmed that PAE significantly enhanced the activity of the p53 signaling pathway（P<0.05）. PPI network analysis indicated that breast cancer type 1 susceptibility protein（BRCA1） and checkpoint kinase 1（CHEK1） were the core down-regulated targets in the p53 pathway. IF verified the high expression of BRCA1 and CHEK1 in LUAD-PDOs and their significant downregulation after PAE intervention（P<0.05）. Furthermore， survival analysis based on The Cancer Genome Atlas（TCGA） database indicated that low expression of BRCA1 and CHEK1 was significantly associated with prolonged overall survival in patients with LUAD（P<0.05）. ConclusionPAE effectively inhibits proliferation of LUAD-PDOs and promotes their apoptosis， its anti-tumor mechanism is potentially associated with the activation of the p53 signaling pathway， with BRCA1 and CHEK1 genes likely serving as key downstream targets for the effects of PAE.

OBJECTIVE: To systematically evaluated clinical efficacy of Kirschner's needle and elastic intramedullary nail fixation in treating proximal humeral fractures in children by Meta-analysis. METHODS: Literature on the treatment of proximal humeral fractures in children with Kirschler needles and elastic intramedullary nails published on Wanfang, VIP, CNKI and China biology medicine (CBM), PubMed, Embase, and Web of Science databases were searched from the establishment of databases to October, 2023. Literature extraction, management and data entry were performed by Endnote X9 and Excel 2019, and Meta-analysis was conducted by RevMan 5.3 software. The operation time, hospital stay, fracture healing time, shoulder joint extension range of motion, disabilities of arm, shoulder and hand(DASH) questionnaire score, Neer score or Constant-Murley score and complications were compared between two groups. RESULTS: A total of 7 literatures were included, 1 was prospective study, 6 were retrospective cohort study. There were 521 children, 264 children in Kirschner wire group and 257 children in elastic intramedullary nail fixation group. The results of Meta analysis showed operation time[WMD=-12.61, 95%CI(-24.89, -0.33), P=0.04], fracture healing time[WMD=-0.26, 95%CI(-0.49, -0.02), P=0.03], total complication rate [OR=6.83, 95%CI(3.33, 14.01), P<0.001], nail tract infection rate[OR=6.77, 95%CI(1.72, 26.69), P=0.006] and displacement fracture rate[OR=3.57, 95%CI(1.35, 9.44), P=0.01] between two groups had statistically differences(P>0.05), while there were no statistically significant difference in comparison of hospital stay, shoulder joint extension range of motion, DASH, Neer score, Constant-Murley score, and incidence of skin irritation between two groups (P>0.05). CONCLUSION Kirschner's needle internal fixation has a short operation time and simple operation, but it has a higher incidence of complications compared with elastic nail internal fixation technique. In terms of efficacy and safety, elastic intramedullary nail fixation is one of the options for the treatment of proximal humeral fractures in children.
Humans ; Fracture Fixation, Intramedullary/instrumentation* ; Child ; Shoulder Fractures/physiopathology* ; Bone Nails ; Bone Wires ; Male ; Needles ; Female

Objective： The aim of this study is to explore the predictive value of biparametric magnetic resonance imaging(bpMRI)for pelvic lymph node metastasis in prostate cancer patients with PSA≤20 μg/L and establish a nomogram. Methods： The imaging data and clinical data of 363 patients undergoing radical prostatectomy and pelvic lymph node dissection in the First Affiliated Hospital of Nanjing Medical University from July 2018 to December 2023 were retrospectively analyzed. Univariate analysis and multivariate logistic regression were used to screen independent risk factors for pelvic lymph node metastasis in prostate cancer, and a nomogram of the clinical prediction model was established. Calibration curves were drawn to evaluate the accuracy of the model. Results： Multivariate logistic regression analysis showed extrocapusular extension (OR=8.08,95%CI=2.62－24.97, P<0.01), enlargement of pelvic lymph nodes (OR=4.45,95%CI=1.16－17.11,P=0.030), and biopsy ISUP grade(OR=1.97,95%CI=1.12－3.46, P=0.018)were independent risk factors for pelvic lymph node metastasis. The C-index of the prediction model was 0.834, which indicated that the model had a good prediction ability. The actual value of the model calibration curve and the prediction probability of the model fitted well, indicating that the model had a good accuracy. Further analysis of DCA curve showed that the model had good clinical application value when the risk threshold ranged from 0.05 to 0.70.Conclusion： For prostate cancer patients with PSA≤20 μg/L, bpMRI has a good predictive value for the pelvic lymph node metastasis of prostate cancer with extrocapusular extension, enlargement of pelvic lymph nodes and ISUP grade≥4.
Humans ; Male ; Prostatic Neoplasms/diagnostic imaging* ; Lymphatic Metastasis ; Retrospective Studies ; Nomograms ; Prostate-Specific Antigen/blood* ; Lymph Nodes/pathology* ; Pelvis ; Predictive Value of Tests ; Prostatectomy ; Lymph Node Excision ; Risk Factors ; Magnetic Resonance Imaging ; Logistic Models ; Middle Aged ; Aged

Acute lung injury (ALI) has been a kind of acute and severe disease that is mainly characterized by systemic uncontrolled inflammatory response to the production of huge amounts of reactive oxygen species (ROS) in the lung tissue. Given the critical role of ROS in ALI, a Fe₃O₄ loaded bovine serum albumin (BSA) nanocluster (BF) was developed to act as a nanomedicine for the treatment of ALI. Combining with NIR irradiation, it exhibited excellent ROS scavenging capacity. Significantly, it also displayed the excellent antioxidant and anti-inflammatory functions for lipopolysaccharides (LPS) induced macrophages (RAW264.7), and Sprague Dawley rats via lowering intracellular ROS levels, reducing inflammatory factors expression levels, inducing macrophage M2 polarization, inhibiting NF-κB signaling pathway, increasing CD4⁺/CD8⁺ T cell ratios, as well as upregulating HSP70 and CD31 expression levels to reprogram redox homeostasis, reduce systemic inflammation, activate immunoregulation, and accelerate lung tissue repair, finally achieving the synergistic enhancement of ALI immunotherapy. It finally provides an effective therapeutic strategy of BF + NIR for the management of inflammation related diseases.

[Objective]To evaluate the effect of polysaccharide from Polygonatum sibiricum on Toll-like receptor 4(TLR4)activation and clarify its antigen specific response-enhancing effect on mice.[Methods]The HEK-BlueTM hTLR4 cell line which was stably transfected with genes of human TLR4 and secreted embryonic alkaline phosphatase(SEAP)was used to quickly evaluate the activation of four different types of polysaccharide from Polygonatum sibiricum,including Polygonatum kingianum NP H2O(PKNH),Polygonatum kingianum AP H2O(PKAH),Polygonatum kingianum NP 5%EtOH(PKNEt)and Polygonatum kingianum AP 5%EtOH(PKAEt)on TLR4.C57BL/10ScNJ(TLR4-/-)mice were used to determine the relationship between the drug effect and TLR4.Ovalbumin(OVA)immunized mouse model was used to evaluate the enhancement of drug on specific immune response.The serum immunoglobulin G1(IgG1)and the cytokines related to the TLR4 signaling pathways such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and interferon-γ inducible protein-10(IP-10)levels were detected by enzyme-linked immunosorbent assay(ELISA),and lymphocyte proliferation was evaluated by 5-[3-(Aarboxymethoxy)phenyl]-3-(4,5-dimethyl-2-thiazolyl)-2-(4-sulfophenyl)-2H-tetrazolium innersalt(MTS)assay.The mRNA expression levels of cytokines related to the TLR4 signaling pathways was determined by reverse transcription-polymerase chain reaction(RT-PCR).[Results]TLR4 was activated most strongly by treatment of neutral polysaccharides from PKNH for 24 h(P<0.001).Compared with wild type,the cell proliferation and the expressions of TNF-α,IL-6 and IP-10 were significantly decreased in PKNH treated splenic lymphocyte from C57BL/10 ScNJ mice(P<0.001).Intragastric administration of PKNH significantly increased serum IgG1 level(P<0.01)and promoted splenic lymphocyte proliferation in OVA immunized mice(P<0.05).[Conclusion]The polysaccharide from Polygonatum sibiricum PKNH enhances the antigen specific immune response through activation of TLR4.

[Objective]To analyze the thought of Professor HE Ruoping in applying Xiaoyao Powder,in order to expand the clinical application of Xiaoyao Powder,and improve the ability of clinical syndrome differentiation and prescription selection.[Methods]Through daily consultations with teachers,collecting medical cases and literature retrieval,this paper discusses the new interpretation of the modification of Xiaoyao Powder.It also presents typical medical cases treated with this formula for conditions such as menstrual disorders,anxiety disorders,ovarian cysts and human papillomavirus(HPV)infection.Furthermore,it summarizes Professor HE's clinical experience in applying Xiaoyao Powder.[Results]Professor HE suggests modifying the original Xiaoyao Powder by removing Mentha haplocalyx Briq.,replacing Zingiber officinale Rosc.with Zingiber officinale Rosc.(dried)to enhance the formula's ability to warm the middle and disperse cold.Additionally,she adds Paeonia suffruticosa Andr.to invigorate blood and dispel blood stasis,thus strengthening the formula's overall effect of promoting blood circulation and tonifying the spleen.Cases treated with Xiaoyao Powder as the base formula such as menstrual disorders,anxiety,ovarian cysts and HPV infections shared the common pathogenesis of liver Qi stagnation and spleen deficiency.By combining differentiation of syndromes with appropriate treatment methods,including regulating and harmonizing Qi and blood,clearing and draining heat and blood stasis,resolving masses and transforming blood stasis,and clearing heat and draining dampness,good therapeutic results were achieved.[Conclusion]Professor HE's clinical application of Xiaoyao Powder often removes Mentha haplocalyx Briq.,and uses Zingiber officinale Rosc.(dried)instead of Zingiber officinale Rosc.added with Paeonia suffruticosa Andr.She believes that this formula can be used for gynecological,psychiatric and other diseases that belong to liver depression and spleen deficiency.Different diseases can be treated with the same formula,expanding the scope of treatment and fully reflecting the role of syndrome differentiation and treatment.It is worth promoting and learning from.

Objective:To analyze the results of Healthy Zhejiang assessment and reveal the weakness in the process of Healthy Zhejiang construction,in order to propose suggestions for promoting the high-quality of Healthy Zhejiang construction.Methods:Descriptive statistics were used to analyze the points-deduction rates of the assessment indicators of Healthy Zhejiang in 90 counties(county-level cities and districts)of Zhejiang Province and non-parametric tests were used to analyze the results of Healthy Zhejiang assessment from 2020 to 2023.Results:The overall effect of Healthy Zhejiang construction from 2020 to 2023 was remarkable,but there were inadequacies and weakness in four aspects:residents'proactive health awareness,primary healthcare service capacity,healthy living environments,and the construction and management of sports facilities.Conclusions:It is still necessary to formulate multifaceted health education program,strengthen the construction of the primary healthcare service system,improve the collection,transportation and disposal system of rural domestic waste,and continuously promote the high-quality development of public sports service.

Objective The myocardial injury was induced by hypobaric hypoxia through regulating the expression of various proteins.The expression of proteins was mainly detected by western blot,but the selection of internal reference proteins and their variations have not been systematically studied.Methods Myocardial injury was induced in a low-pressure,low-oxygen chamber simulating an altitude of 6000 m,for 24 and 72 h.Establishment of the myocardial injury model was confirmed by hematoxylin eosin(HE)staining.Expression levels of internal control proteins,including vinculin,α-tubulin,eukaryotic translation initiation factor-5(EIF5),β-actin,glyceraldehyde-3-phosphate dehydrogenase(GAPDH),cyclophilin B,and cofilin,were detected by Western Blot and total protein expression was detected by Ponceau S and Coomassie Blue staining.An adult mouse cardiomyocytes(AMCMs)injury model was induced by hypoxia for 12 and 24 h and confirmed by terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL staining).Internal control proteins were detected by Western Blot,as in the in vivo model,and total protein expression was detected by Ponceau S and Coomassie Blue staining.Results A myocardial injury model was established by hypobaric hypoxia for 24 and 72 h,the total protein expression levels remained consistent.The expression of internal control proteins including vinculin,EIF5,β-actin,cyclophilin B,and cofilin was consistent between the control and model groups.Expression levels of α-tubulin were similar in the plain control and 24 h hypobaric hypoxia group,but were significantly lower in the 72 h hypobaric hypoxia group compared with the plain control group.GAPDH expression was significantly higher in the 24 and 72 h hypobaric hypoxia groups than in the plain control group.An AMCM injury model was established by hypoxia for 12 and 24 h.Total protein levels and expression levels of the internal control proteins EIF5 and β-actin were consistent,but vinculin,α-tubulin,GAPDH,cyclophilin B,and cofilin expression levels were higher in both hypoxia groups compared with the normoxic control group.Conclusions EIF5 and β-actin may be the suitable loading control proteins for studies of hypobaric hypoxia-induced myocardial injury using Western Blot.Total protein is also a good choice for hypobaric hypoxia studies.

Background: s/Aims: Non-invasive models stratifying clinically significant portal hypertension (CSPH) are limited. Herein, we developed a new non-invasive model for predicting CSPH in patients with compensated cirrhosis and investigated whether carvedilol can prevent hepatic decompensation in patients with high-risk CSPH stratified using the new model. Methods: Non-invasive risk factors of CSPH were identified via systematic review and meta-analysis of studies involving patients with hepatic venous pressure gradient (HVPG). A new non-invasive model was validated for various performance aspects in three cohorts, i.e., a multicenter HVPG cohort, a follow-up cohort, and a carvediloltreating cohort. Results: In the meta-analysis with six studies (n=819), liver stiffness measurement and platelet count were identified as independent risk factors for CSPH and were used to develop the new “CSPH risk” model. In the HVPG cohort (n=151), the new model accurately predicted CSPH with cutoff values of 0 and –0.68 for ruling in and out CSPH, respectively. In the follow-up cohort (n=1,102), the cumulative incidences of decompensation events significantly differed using the cutoff values of <–0.68 (low-risk), –0.68 to 0 (medium-risk), and >0 (high-risk). In the carvediloltreated cohort, patients with high-risk CSPH treated with carvedilol (n=81) had lower rates of decompensation events than non-selective beta-blockers untreated patients with high-risk CSPH (n=613 before propensity score matching [PSM], n=162 after PSM). Conclusions Treatment with carvedilol significantly reduces the risk of hepatic decompensation in patients with high-risk CSPH stratified by the new model.

ObjectiveTo explore effect of Xianlian Jiedu prescription on the recurrence of colorectal cancer （CRC） and investigate the related mechanisms. MethodsA postoperative recurrence model was established in 25 Balb/c mice by injecting CT26 cells subcutaneously into the armpit， followed by surgical removal of 99% of the subcutaneous tumor. The mice were randomly divided into model group， low-dose Xianlian Jiedu prescription （XLJDP-L） group （6.45 g·kg^-1·d^-1）， medium-dose Xianlian Jiedu prescription （XLJDP-M） group （12.9 g·kg^-1·d^-1）， high-dose Xianlian Jiedu prescription （XLJDP-H） group （25.8 g·kg^-1·d^-1）， and 5-fluorouracil （5-FU） group （1×10^-3 g·kg^-1·d^-1）. The mice were euthanized after 14 days of continuous intervention， and recurrent tumor tissue was harvested. Hematoxylin and eosin （HE） staining was used to observe pathological and morphological changes in the recurrent tumor tissue. Immunohistochemistry （IHC） was employed to assess the expression of proliferating cell nuclear antigen （Ki67）， vascular endothelial growth factor （VEGF）， and platelet-endothelial cell adhesion molecule （CD31） in recurrent tumor tissue. The Western blot was used to detect the protein expression levels of angiopoietin-2 （ANG-2）， VEGF， phosphorylated-protein kinase B （p-Akt）， protein kinase B （Akt）， phosphorylated-phosphatidylinositol 3-kinase （p-PI3K）， and phosphatidylinositol 3-kinase （PI3K） in recurrent tumor tissue. ResultsBefore treatment， there were no statistical differences in tumor volume， tumor weight， and body mass among the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group compared to the model group， indicating model stability. After treatment， compared with those in the model group， the tumor volume and tumor weight in the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group were significantly reduced （P<0.01）， showing dose dependency. Meanwhile， there were no significant differences in body weight among the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group compared to the model group. HE staining showed that compared with that in the model group， tumor tissue in the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group had loosely arranged cells， increased intercellular spaces， small and shriveled nuclei， light staining， fewer mitotic figures and atypical nuclei， and increased necrotic areas. IHC showed that compared with those of the model group， the positive rates of Ki67， VEGF， and CD31 in the recurrent tumor tissue of the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group were significantly reduced （P<0.01） in a dose-dependent manner. Western blot results showed that compared with those of the model group， the protein expression levels of ANG-2 and VEGF in the recurrent tumor tissue of the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group were significantly downregulated （P<0.05， P<0.01）， and the p-Akt/Akt and p-PI3K/PI3K ratios were significantly decreased in a dose-dependent manner （P<0.05， P<0.01）. ConclusionXianlian Jiedu prescription significantly inhibits the recurrence of CRC in mice after subcutaneous tumor surgery. The mechanism may involve regulating the PI3K/Akt pathway and downregulating key angiogenic proteins such as ANG-2， VEGF， and CD31.