In this work,a rapid and label-free sensing platform was designed for visual detection of p53 gene.The rolling circle amplification(RCA)process of the assay platform was activated by p53 gene to produce long DNA-wires,which could bound with berberine hydrochloride(BBH)and further enhanced its fluorescence.This method showed high sensitivity with a low detection limit of 5.63 pmol/L,and high specificity toward p53 gene over other interference materials,even for single-base mutation gene.The method could realize the visual detection of targets under the illumination of a UV lamp.In addition,the designed fluorescence detection platform was successfully applied to p53 gene analysis in 10% fetal bovine serum samples,and the relative standard deviation and the recoveries were 0.1% -1.2% and 99.5% -104.7%,respectively.This approach had satisfactory characteristics,such as low cost,label-free,rapidity,high sensitivity,good selectivity and anti-interference ability,and reliable detection capability for complex practical samples,demonstrating a promising prospect in the diagnosis and treatment of diseases,especially for cancer.

In-fusion cloning technology,as a revolutionary and efficient molecular biology tool,has been applied in multiple research fields such as basic biology,biotechnology,and biomedicine.In this article,we introduce a teaching reform project suitable for undergraduate students in the course of"Molecular Bi-ology Laboratory",which utilizes in-Fusion cloning technology to construct a prokaryotic expression vector for alkaline phosphatase mutant genes.Through specific teaching cases,we systematically explored the design and implementation of experimental projects,and focused on analyzing the key and difficult points of the teaching content.Our teaching practice has found that the implementation of this educational re-form project has achieved very good results in enhancing students' core biological literacy,bioinformatics skills,research thinking,and innovation abilities.At the same time,the application of this technology can significantly improve the quality of experimental teaching,providing new ideas and practical refer-ences for promoting the reform and innovation of National First-Class Courses.

With the rapid development of generative artificial intelligence(GAI)technologies,their widespread application in academic research and writing is continuously expanding the boundaries of sci-entific inquiry.However,this trend has also raised a series of ethical and regulatory challenges,inclu-ding issues related to authorship,content authenticity,citation accuracy,and accountability.In light of the growing involvement of AI in generating academic content,establishing an open,controllable,and trustworthy ethical governance framework has become a key task for safeguarding research integrity and maintaining trust within the academic community.This expert consensus outlines ethical requirements across key stages of AI-assisted academic writing-including topic selection,data management,citation practices,and authorship attribution.It aims to clarify the boundaries and ethical obligations surrounding AI use in academic writing,ensuring that technological tools enhance efficiency without compromising in-tegrity.The goal is to provide guidance and institutional support for building a responsible and sustainable research ecosystem.

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.

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.

BACKGROUND:β-Defensin has the ability against influenza A virus and inhibits a series of inflammatory responses induced by influenza A virus infection within cells.There have been no reports on whether hemagglutinin 1 from influenza A virus can induce the secretion of mouse β-defensin and interferon-γ when acting in mouse tracheal epithelial cells.OBJECTIVE:To investigate the effect of recombinant hemagglutinin 1 on the production levels of mouse β-defensin and interferon-γ in mouse tracheal epithelial cells.METHODS:Primary mouse tracheal epithelial cell were divided into six groups:blank control group(Control),recombinant hemagglutinin 1 group(200 ng/mL),recombinant hemagglutinin 1+influenza A virus group,influenza A virus group(2×TCID50),recombinant hemagglutinin 1+inactivated influenza A virus group,and inactivated influenza A virus(I)group.After the mouse tracheal epithelial cells in each group were treated for 4,8,or 24 hours,hematoxylin-eosin staining was used for pathological observation.The mRNA levels of mouse β-defensin 2,3,and 4,and interferon-γ were detected by qRT-PCR.The protein levels of mouse β-defensin 2,3,and 4 were measured by enzyme-linked immunosorbent assay,and the protein expression of interferon-γ was detected by western blot.RESULTS AND CONCLUSION:(1)The recombinant hemagglutinin 1 acting alone or in combination with influenza A virus could cause different pathological changes in tracheal epithelial cells.Phenomena such as vacuolation,nuclear pyknosis and cell fusion could be observed in the cells.(2)Compared with the control group,recombinant hemagglutinin 1 alone or in combination with influenza A virus or inactivated influenza A virus significantly induced the production of mouse β-defensin 2,3,and 4(P<0.05)and interferon-γ in mouse tracheal epithelial cells(P<0.05).These results indicate that recombinant hemagglutinin 1 alone or in combination with influenza A virus can induce the production of mouse β-defensin 2,3,and 4 and interferon-γ in mouse tracheal epithelial cells.

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.

Neurogenic bladder(NB)is one of the most challenging urinary system disorders,with spinal cord injury(SCI)being an important etiological factor.Animal models provide crucial tools for investigating the pathogenesis,therapeutic strategies,and novel drug screening for NB subsequent to SCI.We reviewed and synthesized recent literature on NB animal models after SCI from both domestic and international sources.This review summarizes and analyzes research advancements using these models in terms of animal species,SCI segments,modeling techniques,and evaluation indicators,with the aim of offering insights and guidance for future experimental research based on animal models of NB following SCI.

Neurogenic bladder(NB)is one of the most challenging urinary system disorders,with spinal cord injury(SCI)being an important etiological factor.Animal models provide crucial tools for investigating the pathogenesis,therapeutic strategies,and novel drug screening for NB subsequent to SCI.We reviewed and synthesized recent literature on NB animal models after SCI from both domestic and international sources.This review summarizes and analyzes research advancements using these models in terms of animal species,SCI segments,modeling techniques,and evaluation indicators,with the aim of offering insights and guidance for future experimental research based on animal models of NB following SCI.