ObjectiveBiochemistry and Molecular Biology, a discipline that elucidates life phenomena at the molecular level, serves as a core foundational course in medical education. It provides the theoretical basis for studying other basic and clinical medical subjects, as well as for understanding pathogenesis, disease diagnosis, and treatment. However, its complex content and highly abstract concepts have posed a dual challenge to traditional teaching models: “inefficient instruction” and “inadequate learning outcomes”. Within limited classroom hours, how to engage students and stimulate their intrinsic motivation, and how to help them recognize, understand, and develop a passion for biochemistry from the perspective of the discipline’s essence, have long been key focuses of curriculum research. MethodsUsing the lipid metabolism chapter as an example, this study employs “Rain Classroom”, a generative artificial intelligence (AI)-assisted platform, to support education in four dimensions: teaching, learning, evaluation, and research. In teaching, it assists instructors through virtual experiments, lesson preparation support, knowledge mapping, and assignment design. For learning, it serves as an intelligent study assistant for students, providing automated assignment review, enabling educational resource sharing, and facilitating personalized learning pathways. In evaluation, the platform automates assignment grading, analyzes student performance data, and offers diagnostic feedback and teaching recommendations. In research, it aids educators in collecting and analyzing teaching data, as well as searching for and summarizing relevant literature. ResultsThe results indicate that an educational model integrating teacher-led instruction, student-centered learning, and generative AI assistance significantly enhances teaching quality, students’ self-directed learning abilities, and knowledge mastery. Furthermore, with the support of generative AI, curriculum-based ideological education—focusing on cutting-edge disciplinary advances and topical medical issues—helps cultivate students’ medical spirit of “honoring life and healing the wounded”, thereby fostering the establishment of appropriate professional values. Finally, while generative AI presents both opportunities and challenges for higher education, this study also analyzes potential risks in its teaching applications, emphasizing the need for both instructors and students to avoid over-reliance and to ensure that technological tools consistently serve the fundamental goals of education. ConclusionThis study demonstrates that integrating generative AI, specifically via the “Rain Classroom” platform, can effectively enhance biochemistry education. By supporting teaching, learning, evaluation, and research, this approach improves both educational effectiveness and student outcomes. It also facilitates the incorporation of cutting-edge knowledge and professional ethics, nurturing a patient-centered mindset. Additionally, the study addresses potential implementation risks to ensure that such technological tools remain aligned with the core purpose of education.

Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.

ObjectiveTo establish an objective evaluation system based on the quality characteristics of Gardeniae Fructus without relying on the origin label according to the theory of excellent shape， high quality， and superior effect of authentic medicinal materials， thus providing a scientific basis for the standardization of Gardeniae Fructus. MethodsDifferent batches of Gardeniae Fructus samples were collected from different producing areas. Through herbal textual research and a review of modern literature， fruit length， maximum fruit diameter， and chroma were selected as quantitative indicators to characterize the traditional appearance traits of Gardeniae Fructus. The above indicators were measured by a vernier caliper and a spectrophotometer. Through classical famous prescription data and literature analysis， total iridoid glycosides， total crocus glycosides， and total flavonoids were selected as the core component indicators related to efficacy， and their content was determined. Finally， through cluster analysis， the threshold ranges of the quantitative indicators of excellent shape and high quality were determined， and the applicability of the established evaluation system was verified by the determination of the indicators for each batch of medicinal materials. ResultsThe excellent shape indicators of Gardeniae Fructus were the fruit length-to-maximum fruit diameter ratio ≤1.86， 0°≤h≤45°， and L^*≥ 37.32. The content of total iridoid glycosides， total crocus glycosides， and total flavonoids should be no less than 2.2%， 0.44%， and 0.028%， respectively. The measurement results of different batches of Gardeniae Fructus showed that the evaluation system could effectively distinguish the Gardeniae Fructus samples in line with the traditional cognition of excellent shape and high quality. ConclusionThis study preliminarily constructs an excellent shape and high quality-based quality evaluation system for Gardeniae Fructus， which links appearance traits with chemical indicators and provides a methodological basis for the quality control and scientific grading of this herbal medicine.

Perioperative management， especially the selection and implementation of anesthesia protocols， is increasingly regarded as a potentially modifiable factor affecting long-term postoperative outcomes in tumor patients. This paper systematically discusses the key pathophysiological mechanisms in the perioperative period that influence tumor outcomes， including inflammation and immunosuppression caused by surgical trauma， physiological stress responses， and subsequent changes in the tumor microenvironment， which create opportunities for tumor cell escape and progression. It focuses on the potential effects of different types of anesthetics （sedative-hypnotics， inhaled anesthetics， local anesthetics， analgesics） on the biological behaviors of tumors （such as proliferation， invasion， metastasis， and immune response） as well as their possible molecular mechanisms. Meanwhile， this paper reviews current clinical research evidence on the impact of anesthesia modality selection and specific anesthesia management strategies on the prognosis of tumor patients. Although existing clinical evidence is insufficient to establish standardized perioperative anesthesia protocols that can significantly improve tumor prognosis， both basic and clinical studies strongly suggest that optimizing perioperative management （including the selection of anesthetics， the application of anesthesia techniques， and comprehensive supportive measures） has the potential to influence tumor outcomes by multi-dimensionally regulating the stress-inflammation-immune axis and the tumor microenvironment. Future research should focus on further clarifying the relevant mechanisms and conducting high-quality prospective clinical trials， so as to promote the establishment of evidence-based， precise and individualized perioperative management strategies for tumor patients， and ultimately improve long-term patient survival.

ObjectiveThis study aimed to investigate whether dexmedetomidine （Dex） alleviates stress-induced mammary tumor progression in mice by inhibiting the TLR4/NF‑κB signaling pathway， and to examine its effects on anxiety‑depression‑like behaviors and stress hormone levels. MethodsAn orthotopic EMT6 mammary tumor model was established in mice， and chronic mild stress （CMS） was applied to induce anxiety‑depression‑like states. The mice were divided into six groups： naive， tumor， CMS， CMS+tumor， CMS+tumor+Dex， and CMS+tumor+Vehicle groups. Behavioral changes were evaluated using the open field test and tail suspension test. Plasma stress hormone levels were measured by high‑performance liquid chromatography. Western blot and RT‑qPCR were performed to detect the expression of TLR4， p‑NF‑κB p65， p‑IκBα， and inflammatory cytokines （IL‑6， IL‑1β， TNF‑α） in tumor tissues. In vitro， EMT6 cells were treated with cortisol to mimic the stress-related microenvironment， and the effects of Dex on the TLR4/NF‑κB pathway were assessed by CCK‑8， Western blot， and RT-qPCR. ResultsTumor‑bearing mice exhibited significant anxiety‑depression‑like behaviors （P＜0.05）， along with markedly elevated plasma levels of norepinephrine， epinephrine， and cortisol （P＜0.05）. Chronic stress further aggravated these behavioral abnormalities and promoted tumor growth （P＜0.05）. Dexmedetomidine intervention significantly improved anxiety‑depression‑like behaviors （P＜0.05） and lowered peripheral stress hormone levels （P＜0.05） in stress‑exposed tumor‑bearing mice. In vitro experiments demonstrated that cortisol treatment activated the TLR4/NF‑κB pathway （P＜0.001） and upregulated the expression of inflammatory cytokines （P＜0.05 in EMT6 cells， whereas dexmedetomidine effectively suppressed this activation （P＜0.001）. In the in vivo model， dexmedetomidine treatment downregulated the expression of TLR4， p‑NF‑κB p65， p‑IκBα， and pro‑inflammatory cytokines in tumor tissues （P＜0.001） and significantly inhibited tumor growth （P＜0.05）. ConclusionDexmedetomidine inhibits the TLR4/NF‑κB signaling pathway， alleviates stress‑induced inflammatory responses， improves anxiety- and depression‑ like behaviors， reduces stress hormone levels， and suppresses mammary tumor progression. These findings provide an experimental basis for understanding anxiety- and depression- associated breast tumor progression and suggest that Dexmedetomidine and the TLR4/NF-κB pathway may serve as potential therapeutic targets.

Objective To propose a framework for risk assessment and stepwise management of pulmonary nodules based on existing guidelines and key studies, and to clarify the boundaries for triage, workup escalation, and referral in different detection scenarios. Methods A problem-oriented evidence synthesis approach was used, prioritizing Chinese expert consensus, Fleischner Society Guidelines, British Thoracic Society (BTS) Guidelines, and Lung-RADS. This was supplemented by studies on risk models, imaging measurements, liquid biopsy, tissue sampling, and surgical treatments. This paper is not presented as a formal consensus or a validated clinical pathway. Results A three-tiered framework consisting of basic non-invasive assessment, advanced assessment, and invasive diagnosis was established. It specified the applicable populations, boundaries for the use of risk models, imaging review, management of conflicting results, evaluation of histopathological consistency, and post-intervention follow-up. Liquid biopsy, genetic testing, and artificial intelligence were defined as investigational or adjunctive tools. Conclusion This framework can serve as a preliminary reference for multidisciplinary teams in developing local protocols. It cannot replace applicable guidelines, pathological diagnosis, shared decision-making with patients, or real-world validation.

Abstract In order to explore the pathogenesis of autism spectrum disorder (ASD) from a new perspective, it is found through literature review that vitamin D(Vit D) may play a significant role in the onset and development of ASD by inhibiting early brain overgrowth, regulating metabolism of central nervous system, modulating immune responses, reducing inflammation and influencing genome stability. Furthermore, Vit D supplementation in individuals with ASD has shown to alleviate symptoms. However, the mechanisms by which Vit D affects ASD remains unclear, and the effectiveness of Vit D treatment in improving ASD symptoms remains controversial due to variations in sample sizes, detection methods, dosage and confounding factors. Future research needs to focus on multi center, large scale randomized controlled trials and ASD animal models with Vit D deficiency to further explore the specific role of Vit D in the pathogenesis and treatment of ASD, which could provide reliable evidence for the early detection, intervention and treatment of ASD.

Background and aims:Liver fibrosis is a prevalent pathological stage of various chronic liver diseases and has the potential to progress to liver cirrhosis and hepatocellular carcinoma.However,experimental models for in vivo research are limited.Unexpectedly,increased liver inflammation and fibrosis were previously observed in mice treated with aluminum adjuvant(commercial Imject Alum,a mixture of Al(OH)3 and Mg(OH)2).Our study aimed to reveal the pathogenesis and pathological features of Imject Alum-induced liver injury and evaluate its potential as an experimental model of fibrotic liver disease.Methods and materials:C57BL/6J mice were randomly divided into the following four groups:(ⅰ)control group,which received phosphate-buffered saline injections on days 1,12,26,40,and 54;(ⅱ)Imject Alum(Al(OH)3 160 mg/kg)D26 group,which was administered with Imject Alum(Al(OH)3 160 mg/kg)on days 1,12,and 26;(ⅲ)Imject Alum(Al(OH)3 80 mg/kg)D54;and(ⅳ)Imject Alum(Al(OH)3 160 mg/kg)D54 groups,which were treated with 80 mg/kg and 160 mg/kg of Imject Alum(Al(OH)3),respectively,on days 1,12,26,40,and 54.All reagents were delivered by intraperitoneal injection.Serum biochemical pa-rameters,liver pathology,and expression of genes related to inflammation and fibrogenesis were eval-uated.Transcriptome sequencing was performed.The genetic characteristics of the Imject Alum-induced liver lesions in the existing fibrosis model and patients with cirrhosis were determined.Results:Administration of Imject Alum(Al(OH)3 160 mg/kg)at certain points for 54 days led to extensive hepatic inflammation and fibrosis,accompanied by disturbed bile acid metabolism in mice.Moreover,Imject Alum aggravated liver inflammation and injury by activating the pyroptosis-related inflamma-some pathway.Transcriptome analysis revealed that Imject Alum-induced liver lesions had differentially expressed genes that were significantly enriched in pathways related to inflammation,fibrogenesis,and multiple metabolic processes.Moreover,Imject Alum-induced liver lesions exhibited gene signatures similar to those of existing fibrosis models and patients with cirrhosis.Conclusions:Aluminum adjuvant(Imject Alum;Al(OH)3 160 mg/kg)administration at certain points for 54 days resulted in notable liver injury,inflammation,and fibrosis.This model had similar gene expression characteristics with existing fibrosis models and liver samples from patients with cirrhosis.Overall,aluminum adjuvant(Imject Alum)-induced mouse model may be a novel approach for estab-lishing a liver fibrosis animal model.

Aim To establish and evaluate an alternative meth-od for detecting skin sensitization of cosmetics based on local lymph node assay using bromodeoxyuridine(BrdU)with flow cytometry(FCM).Methods(1)25％hexyl cinnamic alde-hyde(HCA)was chosen as a positive control with an acetone:olive oil(4∶1,V/V,AOO)mixture as a vehicle control for the experiment.The dorsal sides of both ears of mice were treated with test solutions on day 1,day 2,and day 3.Brdu solution was injected inter-peritoneally on day 5.On day 6,the bilateral ears and mandibular lymph nodes were excised,and the number of Brdu positive cells was measured by flow cytometry.The stim-ulation index(SI)was calculated to identify whether it was ≥3,in order to establish the method of LLNA:Brdu-FCM.(2)BrdU-FCM test was conducted using a blind method with the fif-teen reference substances listed in OECD TG429 whose skin sensitization potentials were known.The test substances were dissolved in AOO,N,N-dimethylformamide(DMF)or dimeth-yl sulfoxide(DMSO)at three different concentrations.Tests were performed the same as above.SI and EC2.7 were calculat-ed to evaluate whether the test substance was categorized as a skin sensitizer.The reliability and accuracy of the method were validated by comparing the classification of test substances with that in OECD TG429.Results The SI for 25％HCA was 3.9,showing positive in the skin sensitization test.It demonstrated that the LLNA:Brdu-FCM test method was properly implemen-ted.Nine test substances(2,4-dinitrochlorobenzene,4-pheny-lenediamine,cobalt chloride,2-mercaptobenzothiazole,hexyl-cinnamaldehyde,eugenol,phenyl benzoate,cinnamic alcohol,imidazolidinyl urea)were positive,and six test substances(methyl methacrylate,chlorobenzene,isopropanol,lactic acid,methyl salicylate,salicylic acid)were negative.The method was evaluated with sensitivity of 90％,specificity of 100％,positive prediction rate of 100％,negative prediction rate of 83％,false positive rate of 0％,false negative rate of 17％and accuracy of 93％.The LLNA:BrdU-FCM assay could correctly categorize the test substances that were skin sensitizers or non-sensitizers.Conclusion The LLNA:BrdU-FCM assay appears to be a relia-ble predictor of skin sensitization protential of chemicals,and it is expected to an alternative method for identifying skin sensitization as a supplementary in safety evaluation of cosmetic ingredient.

Objective To study the effect of esketamine on postoperative inflammatory celluar cyto-kines and mental state in the patients with total hysterectomy.Methods A total of 162 patients with elective laparoscopic total hysterectomy in Wenzhou municipal Hospital of Integrated Traditional Chinese and Western Medicine from August 2020 to June 2024 were selected as the study subjects and randomly divided into the control group(n=54),low-dose group(n=53)and high-dose group(n=55).The three groups routinely conducted the general anesthesia.The low-dose group and high dose group were intravenously injected by 0.2 mg/kg and 0.4 mg/kg esketamine once at preoperative 10 min respectively,and the same amount of normal saline in the control group was injected.The general condition and operation time in the three groups,occur-rence of nausea and vomiting,hallucinations,nightmares,agitation,drowsiness and dizziness on postoperative 1 d as well as the VAS scores in 3 groups were recorded.Venous blood on preoperative 1 d(T0),postoperative 1 d(T1)and 3 d(T2)was collected,the levels of the nucleotide-binding oligomerization domain-like receptor pyrin domain protein 3(NLRP3),interleukin(IL)-6,IL-1β,tumor necrosis factor-α(TNF-α),brain-derived neurotrophic factor(BDNF)and neuropeptide Y(NPY)were measured.The Hamilton Anxiety Scale(HA-MA)and Hamilton Depression Rating Scale(HAMD)were used to evaluate the anxiety and depression of the patients.The difference in the HAMA scores generated based on general data at T2 among various the sub-groups between the low-dose group and the control group was analyzed.Results Compared with the control group,the incidence rate of nausea and vomiting and VAS score in the low-dose group and high-dose group were lower,and the difference was statistically significant(P＜0.05).Compared with the control group,the levels of NLRP3,IL-6,IL-1β and TNF-α at T1 and T2 in the low-dose group and high-dose group were lower,and the differences were statistically significant(P＜0.05).Compared with the control group,the BDNF level at T1 and T2 in the low dose group and high dose group was higher,the NPY level was lower,and the differ-ences were statistically significant(P＜0.05).Compared with at T0,the HAMA and HAMD scores of T1 and T2 in the low-dose group and high-dose group were decraesed,and the differences were statistically significant(P＞0.05).Compared with the control group,the HAMA and HAMD scores at T1 and T2 time points the low-dose group and high-dose group were lower,and the differences were statistically significant(P＜0.05).Conclusion Esketamine could improve the anxiety and depression condition and reduce the inflammation level in the patients.