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 establish a stable,reliable,and clinically relevant porcine model of endotoxin-induced acute respiratory distress syndrome(ARDS).Methods Ten 8-month-old male Bama minipigs were deeply sedated,followed by invasive mechanical ventilation and electrocardiographic monitoring.Lipopolysaccharide(LPS)was intravenously pumped at 600 μg/(kg·h)for 3 hours,then maintained at 15 μg/(kg·h)thereafter.Dynamic monitoring was performed at five time points after LPS injection(LPS 0,1,3,5,and 8 h),including arterial blood gas analysis and chest computed tomography(CT)scans.Pathological examination of lung tissues obtained via bronchoscopic biopsy(HE staining and transmission electron microscopy)was conducted.These indicators were comprehensively used to evaluate the success of the animal model.Results At 5 hours after LPS administration,8 minipigs developed symptoms such as skin cyanosis,elevated body temperature,and respiratory distress.The oxygenation index decreased to<300 mmHg.Chest CT scans showed diffuse pulmonary infiltrates.Histopathology revealed alveolar edema and hyaline membrane formation.Transmission electron microscopy demonstrated disruption of pulmonary blood-air barrier,depletion of lamellar bodies in type Ⅱ pneumocytes,inflammatory cell infiltration,and exudation of plasma proteins and fibrin.Compared with LPS 0 h,at LPS 8 h,the oxygenation index and arterial blood pH were significantly decreased(P<0.001),while blood lactic acid and serum potassium were significantly increased(P<0.05);serum calcium and base excess were significantly decreased(P<0.05),and the lung injury score based on HE-stained lung sections was significantly increased(P<0.01).Conclusion The porcine ARDS model established by continuous LPS injection can dynamically simulate the pathophysiological characteristics and typical pathological manifestations of clinical septic ARDS,making it an effective tool to study the pathogenesis,prevention,and treatment strategies of septic ARDS.

Objective:To analyze the clinical value of delayed solidification of cement at freezing point combined with establishment of cement channels in the treatment of further leakage of cement in percutaneous kyphoplasty (PKP).Methods:A retrospective analysis was performed for the medical records of 261 patients with osteoporotic vertebral fracture in the thoracolumbar segment who underwent PKP treatment in Beijing Puren Hospital from April 2019 to April 2023. According to the method of dealing with PKP cement leakage, it was divided into freezing point group (using bone cement combined with cement channel reconstruction treatment at freezing point temperature) and temperature gradient group (using temperature gradient method). There were 128 cases in the freezing point group, including 37 males and 91 females, aged 75.57±4.60 years (range, 65-85 years), and fracture locations were 18 cases in T 10, 30 cases in T 11, 44 cases in T 12, 23 cases in L 1 and 13 cases in L 2. There were 133 cases in the temperature gradient group, including 36 males and 97 females, aged 75.66±4.51 years (range, 65-85 years), and fracture locations were 17 cases in T 10, 32 cases in T 11, 51 cases in T 12, 22 cases in L 1, and 11 cases in L 2. The intraoperative blood loss, operation time, intravertebral cement area, cement leakage area, cement leakage area increase, cement bolus time and incidence of injection difficulty, as well as the pain visual analogue scale (VAS), Oswestry disability index (ODI), kyphosis angle, the height of the anterior edge of the injured vertebral body and the difference between it before and after surgery were compared. Results:All patients were followed up for 3 consecutive months. The intraoperative blood loss and initial cement leakage area were 9.48±2.64 ml and 32.56±7.05 mm 2 in the freezing point group and 9.04±2.25 ml and 32.86±7.00 mm 2 in the temperature gradient group, respectively, and the difference was not statistically significant ( P>0.05) ; The operation time, the area of bone cement in the vertebral body, the final leakage area of bone cement, and the increase of bone cement leakage in the freezing point group were 55.08±4.13 min, 1 175.45±117.11 mm 2, 35.84±8.67 mm 2, and 0.00(0.00, 13.32) mm 2, respectively, and the temperature gradient group were 53.02±3.96 min, 823.70±144.79 mm 2, and 73.38±29.16 mm 2 and 44.39(20.13, 56.61) mm 2, the differences were statistically significant ( P<0.05). The height of the anterior edge of the vertebral body was 21.54±2.06 mm and 21.24±2.33 mm immediately after surgery and 3 months after surgery in the freezing point group, which were higher than those in the temperature gradient group 21.10±1.60 mm and 18.92±1.51 mm, respectively, and the difference was statistically significant ( P<0.05). The VAS scores of the freezing point group were 2.29±0.62 and 1.03±0.66 points, ODI were 23.20%±3.97%, 10.43%±4.33%, and the kyphosis angles were 9.09°±2.80° and 9.44°±2.93°, respectively, which were lower than those of the temperature gradient group (4.11±0.79 and 2.79±0.65 points), ODI (35.97%±6.42%, and 23.73%±5.72%), and the kyphosis angles (10.24°±2.33° and 13.22°±2.56°), the differences were statistically significant ( P<0.05). The operating time of bone cement in the freezing point group was 10.89±2.35 min, which was longer than that in the temperature gradient group 5.77±0.52 min, and the difference was statistically significant ( t=24.021, P<0.001). The incidence of cement injection difficulty was 0 in the freezing point group and 27.1% (36/133) in the temperature gradient group. Conclusion:The establishment of bone cement combined with bone cement channel at freezing point temperature can effectively prolong the bolus time of bone cement and reduce the re-leakage of bone cement, which is conducive to increasing the injection volume and distribution area of bone cement in the vertebral body, effectively reducing the amount of bone cement leakage and obtaining better clinical efficacy.

Objective:To analyze the clinical value of delayed solidification of cement at freezing point combined with establishment of cement channels in the treatment of further leakage of cement in percutaneous kyphoplasty (PKP).Methods:A retrospective analysis was performed for the medical records of 261 patients with osteoporotic vertebral fracture in the thoracolumbar segment who underwent PKP treatment in Beijing Puren Hospital from April 2019 to April 2023. According to the method of dealing with PKP cement leakage, it was divided into freezing point group (using bone cement combined with cement channel reconstruction treatment at freezing point temperature) and temperature gradient group (using temperature gradient method). There were 128 cases in the freezing point group, including 37 males and 91 females, aged 75.57±4.60 years (range, 65-85 years), and fracture locations were 18 cases in T 10, 30 cases in T 11, 44 cases in T 12, 23 cases in L 1 and 13 cases in L 2. There were 133 cases in the temperature gradient group, including 36 males and 97 females, aged 75.66±4.51 years (range, 65-85 years), and fracture locations were 17 cases in T 10, 32 cases in T 11, 51 cases in T 12, 22 cases in L 1, and 11 cases in L 2. The intraoperative blood loss, operation time, intravertebral cement area, cement leakage area, cement leakage area increase, cement bolus time and incidence of injection difficulty, as well as the pain visual analogue scale (VAS), Oswestry disability index (ODI), kyphosis angle, the height of the anterior edge of the injured vertebral body and the difference between it before and after surgery were compared. Results:All patients were followed up for 3 consecutive months. The intraoperative blood loss and initial cement leakage area were 9.48±2.64 ml and 32.56±7.05 mm 2 in the freezing point group and 9.04±2.25 ml and 32.86±7.00 mm 2 in the temperature gradient group, respectively, and the difference was not statistically significant ( P>0.05) ; The operation time, the area of bone cement in the vertebral body, the final leakage area of bone cement, and the increase of bone cement leakage in the freezing point group were 55.08±4.13 min, 1 175.45±117.11 mm 2, 35.84±8.67 mm 2, and 0.00(0.00, 13.32) mm 2, respectively, and the temperature gradient group were 53.02±3.96 min, 823.70±144.79 mm 2, and 73.38±29.16 mm 2 and 44.39(20.13, 56.61) mm 2, the differences were statistically significant ( P<0.05). The height of the anterior edge of the vertebral body was 21.54±2.06 mm and 21.24±2.33 mm immediately after surgery and 3 months after surgery in the freezing point group, which were higher than those in the temperature gradient group 21.10±1.60 mm and 18.92±1.51 mm, respectively, and the difference was statistically significant ( P<0.05). The VAS scores of the freezing point group were 2.29±0.62 and 1.03±0.66 points, ODI were 23.20%±3.97%, 10.43%±4.33%, and the kyphosis angles were 9.09°±2.80° and 9.44°±2.93°, respectively, which were lower than those of the temperature gradient group (4.11±0.79 and 2.79±0.65 points), ODI (35.97%±6.42%, and 23.73%±5.72%), and the kyphosis angles (10.24°±2.33° and 13.22°±2.56°), the differences were statistically significant ( P<0.05). The operating time of bone cement in the freezing point group was 10.89±2.35 min, which was longer than that in the temperature gradient group 5.77±0.52 min, and the difference was statistically significant ( t=24.021, P<0.001). The incidence of cement injection difficulty was 0 in the freezing point group and 27.1% (36/133) in the temperature gradient group. Conclusion:The establishment of bone cement combined with bone cement channel at freezing point temperature can effectively prolong the bolus time of bone cement and reduce the re-leakage of bone cement, which is conducive to increasing the injection volume and distribution area of bone cement in the vertebral body, effectively reducing the amount of bone cement leakage and obtaining better clinical efficacy.

The use of hepatitis B virus (HBV)-positive donor kidneys to expand the donor pool has been implemented, but limited evidence exists regarding their impact on transplant outcomes. This study aimed to investigate the effects of donor HBV infection on transplant outcomes. Methods: Donor and recipient data between 2015 and 2021 were collected. A total of 743 kidney transplant cases were screened, including 94 donor hepatitis B surface antigen (HBsAg)+/recipient HBsAg– (D+R–) and 649 donor HBsAg–/recipient HBsAg– (D–R–) cases. The analysis endpoints included recipient HBV infection, delayed graft function (DGF), peak estimated glomerular filtration rate (eGFR) within 12 months, recipient survival, and death-censored graft survival (DCGS). Results: The D+R– group had a significantly higher risk of HBV infection compared to the D–R– group (6/72 vs. 3/231; relative risk, 6.4; p = 0.007). The risk of HBV transmission decreased significantly with increasing hepatitis B surface antibody (HBsAb) titer (p for trend = 0.003). Furthermore, the D+R– group did not exhibit an increased risk of DGF compared to the D–R– group (odds ratio, 0.70; p = 0.51) in the multivariable mixed model. Both groups had similar peak eGFR within 12 months (β = 1.01, p = 0.71), and this had no impact on patient survival (hazard ratio [HR], 0.36; p = 0.10) and DCGS (HR, 0.79, p = 0.59) in the shared-frailty Cox model. Conclusion: The use of HBsAg-positive donor kidneys appears relatively safe for HBV-immunized recipients in the short term. D+R– does not negatively affect graft function recovery and provides comparable posttransplant outcomes. Maintaining an HBsAb titer over 100 mIU/mL before transplantation is critical to reduce the risk of HBV transmission.

Objective To investigate the effect of microglia activation regulated by C-X3-C motif chemokine ligand 1(CX3CL1)-C-X3-C motif chemokine receptor 1(CX3CR1)pathway on memory function in hemorrhagic shock/resuscitation rats.Methods The experiment was divided into two parts.In the first part,the rats were randomly divided into sham group,model-0.5 hour group,model-1.5 hour group,model-3 hour group,10 rats in each group.There were differences in the time of hemorrhagic shock among each group.In the second part,rats were randomly divided into control group and CX3CL1 group,10 rats in each group.The rats in CX3CL1 group were treated with CX3CL1 protein factor(intraventricular injection),and the rats in control group were treated with saline.All rats were trained in Morris water maze experiments before model construction,and tests of Morris water maze experiments were carried out after 4 days of model construction.After completion,the whole brains were taken for HE staining and immunohistochemical staining.Cerebrospinal fluid was taken for detection of inflammatory cytokines,and hippocampus tissues were taken for Real-time PCR detection and Western blotting detection.Results Compared with the sham group,the escape latency of rats in model group increased,the number of platform crossings and the resident time in the third quadrant decreased.The neuronal state was impaired in HE staining in model group.In addition,compared with the sham group,the expression of ionized calcium binding adaptor molecule-1(Iba1)in the brain of the rats in model group increased,the contents of tumor necrosis factor-α(TNF-α)and interleukin(IL)-6 in the cerebrospinal fluid increased,and the M1-type microglia markers CD16,TNF-α,IL-1β and inducible nitric oxide synthase(iNOS)mRNA content increased.At the same time,compared with the sham group,the expressions of CX3CL1 and CX3CR1 in the brain of model group decreased,and the expressions of phosphorylated nuclear factor-κB(p-NF-κB)and nucleotide binding oligomerization domain(NOD)-like receptor protein 3(NLRP3)increased.However,compared with the control group,rats in CX3CL1 group had reduced escape latency,increased platform crossing times and quadrantⅢresident time,and recovered neuronal states.In addition,the expression of Iba1 in the brain of CX3CL1 group decreased,the contents of TNF-α and IL-6 in the cerebrospinal fluid decreased,the mRNA contents of M1-type microglia markers like CD16,TNF-α,IL-1β and iNOS decreased,and the mRNA contents of markers of M2-type microglia glial like CD206,transforming growth factor-β(TGF-β),arginase-1(Arg1),Chitinase 3-like protein 1(Ym 1)increased.Conclusion CX3CL1 can help inhibit the excessive activation of microglia,induce the polarization of microglia to M2 type,inhibit the polarization of M1 type,reduce the release of inflammatory cytokines,and alleviate the memory function damage induced by hemorrhagic shock/resuscitation.

OBJECTIVE To establish the quality standard of Triadica cochinchinensis and to perform the acute toxicity study. METHODS Appearance properties, powder microscopic identification, and thin-layer chromatography(TLC) identification were researched. The specific chromatogram was established by HPLC. The content of cadmium(Cd), lead(Pb), arsenic(As), copper(Cu), and mercury(Hg) was determined by inductively coupled plasma-mass spectrometry(ICP-MS). Acute toxicity was studied by maximum dose. RESULTS The outer skin of herbs was dark brown, and the inner surface was light yellow brown and fibrous. Besides, crystal sheath fiber was common, and calcium oxalate clusters arranges in rows. In the TLC diagram of the test product, the fluorescent spots of the same color were displayed at the corresponding position of the control product(scopoletin, isofraxidin). Five common peaks were calibrated in the characteristic map and the three characteristic peaks(scopoletin, isofraxidin, dimethylfraxetin) were recognized. The content of the measured heavy metal elements was lower than the national limit standard. The linear correlation coefficient was R2 > 0.999. The precision, stability, repetitive RSD were < 10%. The average recovery rate of the added sample was 80%−120%, and the RSD was < 10%. The maximum dose of the acute toxicity test was 184.09 g·kg−1. The 14 d internal body mass, food intake, organ-body ratios, the serum glutamic pyruvic transaminase, glutamic oxaloacetic transaminase, blood urea nitrogen, and creatinine were not significantly different by comparing with the normal controls. Therefore, no significant toxicity was observed. CONCLUSION The established standard can provide a reference for evaluating the quality of Triadica cochinchinensis. The heavy metal content of ten batches of medicinal materials is within the safe range. Acute toxicity test show that there is no obvious significant adverse teactions after oral administration, and the safe dose range is large, which can provide a reference for the subsequent development and utilization.

Electric vertical takeoff and landing(eVTOL)aircraft was introduced from the aspects of basic concept,development history,classification mode,characteristic and advantage.The feasibility of eVTOL aircraft used in aeromedical rescue was discussed in terms of time,space,transport capacity and safety.It's pointed out that eVTOL aircraft would be applied widely in pre-hospital emergency care,inter-hospital transfer and emergency medical rescue.[Chinese Medical Equipment Journal,2024,45(2):82-86]

Objective To investigate the expression of prolyl 4-hydroxylase β-polypeptide(P4HB)in glioblastoma multiforme(GBM)and its impact on clinical prognosis,as well as on the proliferation and migration of U87 cells.Methods(1)According to the Cancer Genome Atlas(TCGA)database,GTEx database and GEPIA2 database,the difference expression of P4HB in GBM and normal brain tissues were analyzed by R software.(2)A total of 52 patients with GBM who underwent surgical treatment from February 2017 to December 2019 were collected from Department of Neurosurgery,the Second People's Hospital of Guiyang.The normal brain tissues of 10 patients were selected as controls.Immunohistochemical method was used to detect the expression level of P4HB in tumor tissues and normal tissues.The Kaplan-Meier method with the log-rank test was employed for survival analysis.Receiver operating characteristic(ROC)curve was used to analyze the predictive valuable of P4HB expression in survival rate of GBM.Univariate and multivariate Cox regression analysis were used to identify the expression of P4HB and related clinicopathological factors affecting the survival and prognosis of the patients.(3)Human GBM U87 cells were randomly assigned into three groups:control group,NC-siRNA group and P4HB-siRNA group.P4HB expression was interfered with by the transfection of siRNA in P4HB-siRNA group.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the content of P4HB mRNA in U87 cells.Cell counting kit-8(CCK-8)and immunofluorescence assay were used to analyze the effects of P4HB on the proliferation of U87 cells.Scratch test was used to analyze the effects of P4HB on cell migration.Results The expression of P4HB was significantly upregulated in GBM tissues compared with normal brain tissues(P<0.05).The γδ T cells(r=-0.227)and follicular helper T cells(r=-0.226)were negatively correlated with the expression of P4HB,while natural killer cell(r=0.417),macrophages(r=0.374),neutrophils(r=0.344),and immature dendritic cells(r=0.263)were positively correlated with the expression of P4HB.Kaplan-Meier survival analysis showed that the progression-free survival and disease-specific survival of GBM patients with high P4HB expression were significantly lower than those with low expression(P<0.05).ROC curve showed that the area under the curve(AUC)of P4HB in predicting overall survival rate of GBM patients was 0.982,and 1-year,3-year,and 5-year survival was 0.655,0.724,0.861,respectively.The immunohistochemistry results suggested that P4HB protein was significantly highly expressed in GBM tumors.Survival analysis indicated that high expression of P4HB was associated with bad prognosis in GBM patients(P<0.05).Multivariate Cox regression analysis indicated that high expression of P4HB and TERT promoter mutations were the independent prognostic risk factors for GBM(P<0.05).Compared with control group and NC-siRNA group,the expression levels of P4HB were decreased significantly after transfected with siRNA in U87 cells of P4HB-siRNA group(P<0.01),and the proliferation ability and the wound healing rate were decreased significantly in P4HB-siRNA group(P<0.001).Conclusions P4HB is significantly highly expressed in GBM,which indicates that the prognosis of patients is poor.Knockout of P4HB could inhibit cellular proliferation and migration of GBM U87 cells.P4HB may be used as the relevant predictive marker and potential therapeutic target in GBM.