OBJECTIVE To investigate the inhibitory effect of human β defensin-3(HBD3)on the replication of in-fluenza A virus(IAV)[A/PR/8/34(H1N1)virus strain]in human bronchial epithelial cells(BEAS-2B)via the mitochondrial autophagy pathway.METHODS BAS-2B cells were infected with IAV,and cell condition observa-tion:plaque assay and light microscopy.Drug treatment:HBD3,autophagy agonist rapamycin(Rapa),autoph-agy inhibitor LY294002;The expression levels of TUFM、MAVS、NP、M2 and PB1-F2 genes were detected by real-time fluorescence quantitative polymerase chain reaction(qPCR).Western blot was used to detect the protein expression levels of P62、LC3Ⅱ and LC3Ⅰ.RESULTS Plaque formation experiments showed that the number of plaques increased with the increase of viral titer.With the increase of viral titer or the prolongation of infection time,the expres-sion levels of TUFM,MAVS,NP,M2 and PB1-F2 genes in BEAS-2 B cells gradually increased,the expression levels of P62 protein decreased,and the protein expression levels of LC3Ⅱ/LC3Ⅰ increased(P＜0.05).Forty-eight hpi of BE-AS-2B cells with IAV,the number of cells in the IAV group was significantly lower than that in the IAV+HBD3 group,the intercellular space was enlarged,and the cells shrunk significantly.Compared with the IAV group,the expression lev-els of TUFM,MAVS,NP,M2 and PB1-F2 genes in BEAS-2B cells in the IAV+HBD3 group decreased,the expres-sion levels of P62 protein increased,and the protein levels of LC3Ⅱ/LC3Ⅰ decreased(P＜0.05).Gene expression levels of TUFM,MAVS,NP,M2 and PB1-F2 were in the IAV+Rapa+HBD3 group were lower than those in the IAV+Ra-pa group(P＜0.05).The protein expression level of P62 in the IAV+Rapa+HBD3 group was higher than that in the IAV+Rapa group(P＜0.05).The protein expression levels of LC3Ⅱ/LC3Ⅰ were lower in the IAV+Rapa+HBD3 group than in the IAV+Rapa group(P＜0.05).CONCLUSION With the increase of viral titer or the prolongation of in-fection time,the proliferation of IAV in BEAS-2B cells increases,and the cell damage exacerbates.HBD3 can inhibit the replication of IAV after its entry into the cells;HBD3 can protect host cells and inhibit IAV replication by inhibiting MA-VS,TUFM-mediated mitophagy pathways.

Brain computer interface(BCI)is a cutting-edge technology that involves the cross-fertilisation of multiple disci-plines such as neuroscience,computer science,bioengineering,electrical engineering,psychology,etc.It can establish a direct communication path between the brain and external devices,providing the possibility for patients with spinal cord injury(SCI),stroke,amyotrophic lateral sclerosis and other conditions to restore their motor or communication abilities.From the first re-cording of brain electrical activity by Hans Berger,to the gradual development of BCI technology,it shows the continuous pro-gress of BCI technology.According to the signal acquisition method,BCI can be classified as noninvasive and invasive types,and the application scenarios of the two experimental paradigms are different.This review explores the current applications of BCI in spinal cord injury,including the treatment of paraplegic patients and the relief of neuropathic pain.BCI devices will develop to-wards the directions of personalization,household use and intelligence,and BCI technology is expected to demonstrate its unique value and potential in more fields.

Objective:To learn about the clinical characteristics of patients with neurobrucellosis (NB) and provide references for clinical diagnosis and treatment of NB.Methods:The clinical data of 22 NB patients diagnosed and treated at Beidahuang Industry Group General Hospital from January 2018 to November 2024 and 178 healthy individuals undergoing physical examinations during the same period were retrospectively collected. The epidemiological characteristics, clinical manifestations, laboratory tests, treatment and prognosis of NB patients were analyzed.Results:Among the 22 NB patients, 12 were males (54.55%) and 10 were females (45.45%). The age was (51.77 ± 12.75) years old, ranging from 27 to 80 years old. Most of the patients were farmers (95.45%, 21/22), and 16 cases (72.73%) had contacted with cattle/sheep. The onset seasons were mainly in summer (40.91%, 9/22) and spring (31.82%, 7/22). Among all NB patients, there were 10 cases of encephalitis/meningoencephalitis, 9 cases of myelitis, and 3 cases of meningitis. The general symptoms were mainly fever (68.18%, 15/22), the neurological symptoms were mainly nausea and vomiting (36.36%, 8/22), and the physical signs were mainly muscle weakness (50.00%, 11/22) and pathological signs (45.45%, 10/22). The laboratory test results showed that the levels of hemoglobin, hematocrit, C-reactive protein, total protein, albumin, γ-glutamyl transpeptidase, α-hydroxybutyric acid dehydrogenase, lactate dehydrogenase, and glutathione reductase in NB patients were significantly different from those in healthy individuals ( P < 0.001). Brain magnetic resonance imaging revealed ischemic changes in 5 cases (22.73%), abnormal brain signals in 2 cases (9.09%), and demyelinating lesions in white matter in 1 case (4.55%). After treatment, 18 NB patients were followed up and showed good prognosis, with only 2 cases exhibiting varying degrees of sequelae (walking disorders or memory impairment). Conclusions:The clinical manifestations of NB patients are diverse. A comprehensive judgment should be made by combining epidemiological characteristics, clinical manifestations, laboratory tests and imaging examinations.

Purpose To investigate the immunohistochemical HER2 expression in a large group of patients with urothelial carcinoma and to compare the results with the pathologic features and survival rate.Methods A total of 519 urothelial carcinoma specimens were collected from two centers.HER2 expression was measured by EnVision immuno-histochemistry.HER2 expression was compared with clinicopathological parameters,and further analyzed in relation to patient prognosis.Results The median age of the 519 patients was 66 years with a male to female ratio of 1.93∶1.Among them,160 cases were radical specimens,and 359 were transurethral resection specimens.The overall HER2 positivity rate was 61.7％(320/519),of which 148 cases(28.5％)were HER2 1+,238(45.9％)were HER2 2+,and 82(15.8％)were HER2 3+.In addition,51 cases(9.8％)were HER2-negative.HER2-positive ex-pression was associated with age,tumor location,histological grade,histological type,surgical approach,lymphovas-cular invasion,and neural invasion(all P＜0.05),but there was no significant correlation with gender,pT stage,muscular invasion,or lymph node metastasis.Univariate and multivariate logistic regression analysis showed that age≥ 66 years,higher tumor grade,and lymphovascular invasion were risk factors for positive HER2 expression,and high histological grade and lymphovascular invasion were independent risk factors affecting HER2 expression after controlling for confounders.Survival analysis showed no significant difference in recurrence-free survival between patients with HER2-positive and HER2-negative non-muscle-invasive urothelial carcinoma(P=0.274).Conclusion High histologic grade,tumor lymphovascular invasion,and neural invasion were associated with positive HER2 expression in patients with urothelial carcinoma,and higher histologic grade and lymphovascular invasion are important factors affect-ing HER2 expression.However,HER2-positive expression may not affect the prognosis of patients with non-muscle-invasive bladder urothelial carcinoma.

OBJECTIVE To investigate the inhibitory effect of human β defensin-3(HBD3)on the replication of in-fluenza A virus(IAV)[A/PR/8/34(H1N1)virus strain]in human bronchial epithelial cells(BEAS-2B)via the mitochondrial autophagy pathway.METHODS BAS-2B cells were infected with IAV,and cell condition observa-tion:plaque assay and light microscopy.Drug treatment:HBD3,autophagy agonist rapamycin(Rapa),autoph-agy inhibitor LY294002;The expression levels of TUFM、MAVS、NP、M2 and PB1-F2 genes were detected by real-time fluorescence quantitative polymerase chain reaction(qPCR).Western blot was used to detect the protein expression levels of P62、LC3Ⅱ and LC3Ⅰ.RESULTS Plaque formation experiments showed that the number of plaques increased with the increase of viral titer.With the increase of viral titer or the prolongation of infection time,the expres-sion levels of TUFM,MAVS,NP,M2 and PB1-F2 genes in BEAS-2 B cells gradually increased,the expression levels of P62 protein decreased,and the protein expression levels of LC3Ⅱ/LC3Ⅰ increased(P＜0.05).Forty-eight hpi of BE-AS-2B cells with IAV,the number of cells in the IAV group was significantly lower than that in the IAV+HBD3 group,the intercellular space was enlarged,and the cells shrunk significantly.Compared with the IAV group,the expression lev-els of TUFM,MAVS,NP,M2 and PB1-F2 genes in BEAS-2B cells in the IAV+HBD3 group decreased,the expres-sion levels of P62 protein increased,and the protein levels of LC3Ⅱ/LC3Ⅰ decreased(P＜0.05).Gene expression levels of TUFM,MAVS,NP,M2 and PB1-F2 were in the IAV+Rapa+HBD3 group were lower than those in the IAV+Ra-pa group(P＜0.05).The protein expression level of P62 in the IAV+Rapa+HBD3 group was higher than that in the IAV+Rapa group(P＜0.05).The protein expression levels of LC3Ⅱ/LC3Ⅰ were lower in the IAV+Rapa+HBD3 group than in the IAV+Rapa group(P＜0.05).CONCLUSION With the increase of viral titer or the prolongation of in-fection time,the proliferation of IAV in BEAS-2B cells increases,and the cell damage exacerbates.HBD3 can inhibit the replication of IAV after its entry into the cells;HBD3 can protect host cells and inhibit IAV replication by inhibiting MA-VS,TUFM-mediated mitophagy pathways.

Brain computer interface(BCI)is a cutting-edge technology that involves the cross-fertilisation of multiple disci-plines such as neuroscience,computer science,bioengineering,electrical engineering,psychology,etc.It can establish a direct communication path between the brain and external devices,providing the possibility for patients with spinal cord injury(SCI),stroke,amyotrophic lateral sclerosis and other conditions to restore their motor or communication abilities.From the first re-cording of brain electrical activity by Hans Berger,to the gradual development of BCI technology,it shows the continuous pro-gress of BCI technology.According to the signal acquisition method,BCI can be classified as noninvasive and invasive types,and the application scenarios of the two experimental paradigms are different.This review explores the current applications of BCI in spinal cord injury,including the treatment of paraplegic patients and the relief of neuropathic pain.BCI devices will develop to-wards the directions of personalization,household use and intelligence,and BCI technology is expected to demonstrate its unique value and potential in more fields.

Purpose To investigate the immunohistochemical HER2 expression in a large group of patients with urothelial carcinoma and to compare the results with the pathologic features and survival rate.Methods A total of 519 urothelial carcinoma specimens were collected from two centers.HER2 expression was measured by EnVision immuno-histochemistry.HER2 expression was compared with clinicopathological parameters,and further analyzed in relation to patient prognosis.Results The median age of the 519 patients was 66 years with a male to female ratio of 1.93∶1.Among them,160 cases were radical specimens,and 359 were transurethral resection specimens.The overall HER2 positivity rate was 61.7％(320/519),of which 148 cases(28.5％)were HER2 1+,238(45.9％)were HER2 2+,and 82(15.8％)were HER2 3+.In addition,51 cases(9.8％)were HER2-negative.HER2-positive ex-pression was associated with age,tumor location,histological grade,histological type,surgical approach,lymphovas-cular invasion,and neural invasion(all P＜0.05),but there was no significant correlation with gender,pT stage,muscular invasion,or lymph node metastasis.Univariate and multivariate logistic regression analysis showed that age≥ 66 years,higher tumor grade,and lymphovascular invasion were risk factors for positive HER2 expression,and high histological grade and lymphovascular invasion were independent risk factors affecting HER2 expression after controlling for confounders.Survival analysis showed no significant difference in recurrence-free survival between patients with HER2-positive and HER2-negative non-muscle-invasive urothelial carcinoma(P=0.274).Conclusion High histologic grade,tumor lymphovascular invasion,and neural invasion were associated with positive HER2 expression in patients with urothelial carcinoma,and higher histologic grade and lymphovascular invasion are important factors affect-ing HER2 expression.However,HER2-positive expression may not affect the prognosis of patients with non-muscle-invasive bladder urothelial carcinoma.

BACKGROUND:In recent years,the application of exosomes of periodontal ligament stem cells in periodontal tissue regeneration engineering has been widely studied,but the effect of exosomes on periodontal ligament stem cells derived from inflammatory environment is still unclear.OBJECTIVE:To investigate the effects of exosomes secreted by periodontal ligament stem cells from healthy and inflammatory environments on the proliferation and differentiation of periodontal ligament stem cells from inflammatory environments.METHODS:Human periodontal ligament stem cells from healthy and inflammatory tissues were isolated and cultured by enzyme digestion method.Exosomes were extracted from two kinds of periodontal ligament stem cells using ultracentrifugation.Passage 3 periodontal ligament stem cells derived from inflammatory tissue were selected and cultured in three groups.Cells in the blank group were cultured routinely.The healthy exosome group was added with exosomes secreted by peripheral ligament stem cells derived from healthy tissue.The inflammatory exosome group was added with exosomes secreted by human periodontal ligament stem cells derived from inflammatory tissue.Cell proliferation and cloning were detected.The expression of alkaline phosphatase,the formation of mineralized nodules,and the expression of mRNA and protein of genes related to osteogenesis were detected under osteogenic differentiation.RESULTS AND CONCLUSION:(1) CCK-8 assay and clonal formation test showed that compared with the blank group,two kinds of exosomes could promote the proliferation and colony formation of periodontal ligament stem cells from inflammatory tissue (P<0.05),and the effect of the healthy exosome group was stronger than that of the inflammatory exosome group (P<0.05).(2) Alkaline phosphatase and alizarin red staining showed that compared with the blank group,the two kinds of exosomes could promote the expression of alkaline phosphatase and the formation of mineralized nodules in periodontal ligament stem cells from inflammatory tissue,and the promoting effect of the healthy exosome group was stronger than that of the inflammatory exosome group.RT-PCR and western blot assay showed that compared with the blank group,the two kinds of exosomes could promote the expression of alkaline phosphatase,RUNX2,and type Ⅰ collagen mRNA and protein in periodontal ligament stem cells from inflammatory tissue (P<0.05).The promoting effect of the healthy exosome group was stronger than that of the inflammatory exosome group (P<0.05).(3) The results showed that exosomes secreted by human periodontal ligament stem cells could promote the proliferation and osteogenic differentiation of periodontal ligament stem cells derived from inflammatory environments,and the promoting effect of exosomes secreted by human periodontal ligament stem cells derived from healthy tissues was better than that from human periodontal ligament stem cells derived from inflammatory tissues.

Objective:To compare the the cultivation effects of human embryos in dry and humid incubators.Methods:A total of 479 infertile patients who underwent in vitro fertilization (IVF) treatment at Reproductive Center of Guiyang Maternal and Child Health Care Hospital from October 2020 to April 2022. The study was divided into two stages. The first stage of the study was a self-comparative research with 95 cases from the same period and source. The embryos were divided into dry and humid incubator groups to compare the embryo development indicators. In the second stage of the study, the patients were divided into six groups, including 10 μL humid incubator group ( n=64), 20 μL humid incubator group ( n=64), 30 μL humid incubator group ( n=64), 10 μL dry incubator group ( n=64), 20 μL dry incubator group ( n=64), and 30 μL dry incubator group ( n=64). The general clinical data, embryo development indicators, pregnancy outcomes, and the osmotic pressure and pH values of each group at 24 h, 48 h and 72 h were detected and compared. Results:After cultivation of the same patient's embryos in dry and humid incubator, the total blastocyst formation rate [62.3% (162/260)] and high-quality blastocyst rate [24.6% (64/260)] in dry incubator were lower than those in the humid incubator [71.6% (252/352), P=0.015; 32.1% (113/352), P=0.043]. Compared with the other microdroplet groups, the osmotic pressure of cleavage culture medium in 10 μL group of dry incubator at 48 h and 72 h and blastocyst culture medium were significantly increased, the differences among the groups were significant (cleavage culture medium, all P<0.001; blastocyst culture medium, P=0.006, P=0.008). There was no significant difference in pH value among different microdroplet volume groups at the same period (all P>0.05). There were no significant differences in general data among the different microdroplet groups (all P>0.05). Compared with the other microdroplet groups, 10 μL dry incubator group exhibited significantly lower transferable embryo rate (all P<0.001). When compared with 20 μL and 30 μL groups in both dry and humid incubators, 10 μL dry incubator group showed a lower day 5 blastocyst formation rate, lower total blastocyst formation rate, and lower high-quality blastocyst formation rate, the differences among the groups were significant (all P<0.05). There were no significant differences in the number of transferred embryos, the ratio of cleavage-stage embryos and the ratio of high-quality embryos among different groups (all P>0.05). Compared with the other microdroplet groups, the clinical pregnancy rate, the embryo implantation rate, the live birth rate of fresh transplanted embryos and the cumulative pregnancy rate in 10 μL group in the dry incubator decreased, and the miscarriage rate increased, but all were not significant (all P>0.05). Conclusion:Compared with humid incubators, there are no significant differences in embryo development and pregnancy outcomes for droplet volumes of 20 μL or above in dry incubators. However, the 10 μL microdroplet culture in the dry incubator is not conducive to embryonic development, which may be related to the increased osmotic pressure of the microdroplet.

Objective:This study aimed to evaluate the feasibility and safety of surgical intervention for patients with stage Ⅳ esophageal cancer who demonstrated tumor regression following first-line treatment.Methods:This was a descriptive case series. The inclusion criteria for surgery were as follows: (1) an initial diagnosis of stage Ⅳ esophageal cancer, i.e. cT4b or cM1; (2) the presence of residual tumor following first-line therapy deemed potentially resectable upon reassessment; and (3) sufficient organ function to tolerate surgical procedures. Clinical data were retrospectively collected for 63 patients with stage Ⅳ esophageal cancer who underwent surgery following first-line therapy at Sun Yat-sen University Cancer Center between January 2014 and December 2023. Of these patients, 12 were initially staged as IVA, and 51 as IVB. Post-treatment restaging revealed that 9 patients achieved a clinical complete response, while 3 were downstaged to stage Ⅰ, 14 to stage Ⅱ, 24 to stage Ⅲ, and 13 to stage ⅣB (with regression of distant metastatic lesions enabling curative resection). Surgical approaches included right thoracic esophagectomy ( n=55), left thoracic esophagectomy ( n=4), and transmediastinal esophagectomy ( n=4). Additionally, 7 patients required extended organ resection. Two-field lymph node dissection was performed in 49 patients, while 14 underwent three-field lymph node dissection. Postoperative management varied: 31 patients received no adjuvant therapy, 11 underwent immunochemotherapy, 8 received immunotherapy alone, 8 underwent chemotherapy, 4 received chemoradiotherapy, and 1 received combined radiotherapy and immunotherapy. The primary endpoints were overall survival (OS) and progression-free survival (PFS), with secondary endpoints including surgical outcomes and postoperative complications. Results:All 63 patients successfully underwent surgery without intraoperative mortality. R0 resection was achieved in 58 cases (92.1%), while R1 and R2 resections were performed in 1 case (1.6%) and 4 cases (6.3%), respectively. The mean operative time was 357±135 minutes. Postoperative complications were observed in 27 cases (42.9%), with 9 cases (14.3%) classified as Clavien-Dindo grade Ⅲ or Ⅴ. One patient (1.6%) died perioperatively. The median follow-up duration was 21 months (range: 4–107 months). The median OS was 64.8 months (95% CI: 50.9–78.6 months), and the median PFS was 68.0 months (95% CI: 53.9–82.3 months). Among 24 patients with supraclavicular lymph node metastases, 6 experienced recurrence and 8 died. Of 25 patients with abdominal metastases, 3 had recurrence and subsequently died. All 4 patients with lung metastases and both patients with bone metastases experienced recurrence and death.Conclusions:Surgical intervention is a feasible and safe treatment option for selected patients with stage Ⅳ esophageal cancer who demonstrate the potential for curative resection following first-line therapy.