Objective To establish a sandwich enzyme-linked immunosorbent assay (ELISA) method for the quantitative detection of Chromogranin A (CHGA) in saliva, and to explore its preliminary application in the testing of saliva samples. Methods Recombinant human CHGA protein was used to immunize BALB/c mice, and monoclonal antibodies (mAbs) were prepared and screened using conventional hybridoma technology. A double-antibody sandwich ELISA detection method was constructed, and the matrix effect of saliva samples was optimized. This method was then applied to detect the concentration of CHGA in the saliva of stressed individuals. Results Twenty-one stable hybridoma cell lines secreting high affinity anti-human CHGA antibodies were obtained. A pair of detection antibodies with the best effect was selected, and the optimal coating concentration was determined to be 10 μg/mL, with the optimal dilution of detection antibodies being 1:32 000. The accuracy and reproducibility of this method were verified, with both intra-batch and inter-batch variation coefficients less than 15×, and the recovery rate between 80× and 120×. The matrix effect was further optimized to make it suitable for saliva sample detection. Saliva samples from individuals in different stress states were collected, and the CHGA levels were detected using the method established in this study, indicating its potential to reflect the intensity of stress. Conclusion A reliable saliva CHGA ELISA detection method has been successfully established, and its potential as a biomarker in stress-related research has been preliminarily explored.
Saliva/metabolism* ; Enzyme-Linked Immunosorbent Assay/methods* ; Humans ; Animals ; Mice, Inbred BALB C ; Mice ; Chromogranin A/immunology* ; Antibodies, Monoclonal/immunology* ; Female ; Male ; Reproducibility of Results ; Adult

Advances in targeted therapy and immunotherapy have significantly improved clinical outcomes for patients with advanced non-small cell lung cancer (NSCLC), reshaping treatment paradigms. However, most patients ultimately face drug resistance, with limited options for subsequent therapies and suboptimal treatment efficacy, presenting a prominent challenge in current clinical practice. Antibody-drug conjugates (ADCs), characterized by high efficacy and favorable safety profiles, have emerged as a promising therapeutic frontier in recent years. This systematic review provides a comprehensive overview of the latest advancements in ADCs-based therapies for lung cancer, alongside discussions of the prevailing challenges in this rapidly evolving domain.
.
Humans ; Carcinoma, Non-Small-Cell Lung/immunology* ; Immunoconjugates/therapeutic use* ; Lung Neoplasms/immunology* ; Antineoplastic Agents/therapeutic use* ; Immunotherapy

Mutations in ion channel genes have long been implicated in a spectrum of epilepsy syndromes. However, therapeutic decision-making is relatively complex for epilepsies associated with channelopathy. Therefore, in the present study, we used a patient-derived organoid model with a novel SCN2A mutation (p.E512K) to investigate the potential of utilizing such a model as a platform for preclinical testing of anti-seizure compounds. The electrophysiological properties of the variant Nav1.2 exhibited gain-of-function effects with increased current amplitude and premature activation. Immunofluorescence staining of patient-derived cortical organoids (COs) displayed normal neurodevelopment. Multielectrode array (MEA) recordings of patient-derived COs showed hyperexcitability with increased spiking and remarkable network bursts. Moreover, the application of patient-derived COs for preclinical drug testing using the MEA showed that they exhibit differential responses to various anti-seizure drugs and respond well to carbamazepine. Our results demonstrate that the individualized organoids have the potential to serve as a platform for preclinical pharmacological assessment.
Organoids/physiology* ; NAV1.2 Voltage-Gated Sodium Channel/genetics* ; Humans ; Anticonvulsants/pharmacology* ; Epilepsy/drug therapy* ; Mutation ; Cerebral Cortex/drug effects* ; Action Potentials/drug effects* ; Carbamazepine/pharmacology*

Objective To analyze the characteristics and drug resistance of pathogenic bacteria isolated from patients undergoing solid organ transplantation(SOT)at West China Hospital,Sichuan University in re-cent years,in order to provide a basis for empirical anti infective treatment after SOT surgery.Methods A retrospective analysis was conducted on the isolation of major pathogens and their resistance to common anti-biotics from various specimens collected from patients undergoing kidney transplantation(KT),liver trans-plantation(LTx),and lung transplantation(LT)at West China Hospital,Sichuan University from 2017 to 2022.Results A total of 1 077 non-duplicate strains of pathogens were isolated from the samples of patients with infections after SOT surgery during the 6-year period,of which approximately 74.8％(806/1 077)were Gram negative bacteria and 25.2％(271/1 077)were Gram positive bacteria.There were differences in the distribution of pathogenic bacteria among different types of SOT groups and different specimens.Compared to E.coli isolated from urine specimens,the strains of E.coli isolated from non-urinary specimens exhibited a higher resistance rate to common antimicrobial drugs(P＜0.05).The resistance rate of E.coli to β-lactam/β-lactamase inhibitor combinations(cefoperazone/sulbactam and piperacillin/tazobactam)in the LTx group was significantly higher than that in the KT group(P＜0.05).The overall proportion of multidrug-resistant bac-teria after SOT surgery was 11.3％(122/1 077).The proportion of carbapenem resistant Acinetobacter bau-mannii and carbapenem resistant Pseudomonas aeruginosa among the same group and type of pathogens in the LTx group(93.8％,37.5％)was significantly higher than that in the KT group(55.8％,9.2％).Conclusion The specimen types and strain distribution of pathogenic bacteria after different types of SOT surgery are different.The same pathogenic bacteria have different antibiotic resistance among different types of SOT groups and specimens.Therefore,it is necessary to strengthen the pathogen examination after different types of SOT and optimize the anti infection treatment plan related to transplantation based on drug sensitivity re-sults.

Objective: To establish a mouse model of lung injury induced by self-antigens and explore its pathological mechanisms to provide a reliable animal model for studying the pathogenesis of rheumatoid arthritis-associated interstitial lung disease(RA-ILD). Methods: The mice were injected intradermal with antigenemulsion made of complete freund′s adjuvant(CFA) and lung tissue protein, and the emulsion prepared with incomplete Freund′s adjuvant(IFA) was used to enhance immunity. HE staining was used to observe the histopathological changes in the mice. Masson staining was used to detect pulmonary fibrosis. The mRNA levels of rheumatoid factor(RF), krebs von den lungen-6(KL-6) and surfactant protein D(SP-D) were detected by qPCR. The levels of ACPA, IgG1, IgG2a and IgG3 antibodies were detected by ELISA. The changes of inflammatory cells and α-smooth muscle actin(α-SMA)expression in lung tissue were detected by immunofluorescence staining. The protein expression levels of Collagen I and α-SMA were detected by Western blot. The changes of immune cells were detected by flow cytometry. Results: HE staining showed that inflammatory cell infiltration increased and tissue structure changed significantly in lung tissue after the model was established by self-lung tissue antigen. Masson staining showed increased collagen deposition in lung tissue of model mice. qPCR tests revealed elevated mRNA levels of RF, KL-6 and SP-D. ELISA tests revealed elevated levels of ACPA, IgG1 and IgG3 antibodies. Immunofluorescence results showed that monocytes and T cells increased, and α-SMA expression increased in the model group. Western blot results showed increased protein expressions of Collagen Ⅰ and α-SMA. The flow cytometry results showed an increase in T cells and monocytes in the lung tissue. Conclusion The mouse model of lung injury induced by self-antigens is successfully established, and T cells and monocytes may be involved in the occurrence and progression of the disease.

Objective: To explore the role of tumour necrosis factor-α(TNF-α) in splenic infection of mice by Leishmania major. Methods: To establish an infection model, promastigotes of Leishmania were injected intradermally into the right hind foot of mice. The thickness of the footpad and body weight were measured to monitor the infection. Histological changes in the spleen after infection were observed by HE staining. Changes in lymphocytes and monocytes in the spleen were detected by flow cytometry. The expression level of arginase-1(Arg-1) and inducible nitric oxide synthase(iNOS) in the spleen was determined by indirect immunofluorescence. The effect of TNF-α on macrophage infection with Leishmania was evaluated in vitro. Results: Compared to B6.WT mice, the spleens of B6.TNF-/- mice showed significant enlargement 42 days post-infection, with structural disruption. Various cells infiltrated and were dispersed throughout the entire spleen. Flow cytometry results indicated that after infection with Leishmania, there was no significant change in the proportions of T cells and B cells in the spleens of the mice, while CD11b monocytic cells significantly increased. Immunofluorescence results revealed that the M2 macrophage/monocyte marker Arg-1 was highly expressed in the spleens of B6.TNF-/- mice(P < 0. 05) . The expression of iNOS in the spleens of B6. WT mice was relatively strong (P < 0. 05) . In vitro studies found that the absence or inhibition of TNF⁃α significantly increased the infection of Leishmania by peritoneal macrophages ( P <0. 01), while the addition of TNF-a markedly inhibited this infection (P <0. 01). Conclusion The splenic infec-tion in B6. TNF mice following subcutaneous inoculation of Leishmania in the hind footpad may be associatedwith the absence of TNF-a, which leads to M2-type differentiation of macrophages and reduced nitric oxide producton.

In March 2020, the outflow of age limited videos from ICU in Spain inspired us to rethink whether there is age discrimination in the allocation of scarce medical resources. This paper frist reflected on the problem of age discrimination caused by this phenomenon from four moral intuitions: the sacred view of life, the quality of life and values, public health ethics and Chinese culture, and then examined whether it is illegal from the legal level, finally pointed out the negative impact on the society, and put forward that taking age as the standard for the allocation of scarce medical resources is not suitable for China’s national conditions.

The Spt-Ada-Gcn5-acetyltransferase (SAGA) is an ancillary transcription initiation complex which is highly conserved. The ADA1 (alteration/deficiency in activation 1, also called histone H2A functional interactor 1, HFI1) is a subunit in the core module of the SAGA protein complex. ADA1 plays an important role in plant growth and development as well as stress resistance. In this paper, we performed genome-wide identification of banana ADA1 gene family members based on banana genomic data, and analyzed the basic physicochemical properties, evolutionary relationships, selection pressure, promoter cis-acting elements, and its expression profiles under biotic and abiotic stresses. The results showed that there were 10, 6, and 7 family members in Musa acuminata, Musa balbisiana and Musa itinerans. The members were all unstable and hydrophilic proteins, and only contained the conservative SAGA-Tad1 domain. Both MaADA1 and MbADA1 have interactive relationship with Sgf11 (SAGA-associated factor 11) of core module in SAGA. Phylogenetic analysis revealed that banana ADA1 gene family members could be divided into 3 classes. The evolution of ADA1 gene family members was mostly influenced by purifying selection. There were large differences among the gene structure of banana ADA1 gene family members. ADA1 gene family members contained plenty of hormonal elements. MaADA1-1 may play a prominent role in the resistance of banana to cold stress, while MaADA1 may respond to the Panama disease of banana. In conclusion, this study suggested ADA1 gene family members are highly conserved in banana, and may respond to biotic and abiotic stress.
Musa/genetics* ; Phylogeny ; Fungal Proteins ; Cell Nucleus ; Histones ; Stress, Physiological/genetics*

Objective To establish a nomogram model for predicting clinical pregnancy rate in patients with endometriosis undergoing fresh embryo transfer.Methods We retrospectively collected the data of 464 endometriosis patients undergoing fresh embryo transfer,who were randomly divided into a training dataset(60%)and a testing dataset(40%).Using univariate analysis,multiple logistic regression analysis,and LASSO regression analysis,we identified the factors associated with the fresh transplantation pregnancy rate in these patients and developed a nomogram model for predicting the clinical pregnancy rate following fresh embryo transfer.We employed an integrated learning approach that combined GBM,XGBOOST,and MLP algorithms for optimization of the model performance through parameter adjustments.Results The clinical pregnancy rate following fresh embryo transfer was significantly influenced by female age,Gn initiation dose,number of assisted reproduction cycles,and number of embryos transferred.The variables included in the LASSO model selection included female age,FSH levels,duration and initial dose of Gn usage,number of assisted reproduction cycles,retrieved oocytes,embryos transferred,endometrial thickness on HCG day,and progesterone level on HCG day.The nomogram demonstrated an accuracy of 0.642(95%CI:0.605-0.679)in the training dataset and 0.652(95%CI:0.600-0.704)in the validation dataset.The predictive ability of the model was further improved using ensemble learning methods and achieved predicative accuracies of 0.725(95%CI:0.680-0.770)in the training dataset and 0.718(95%CI:0.675-0.761)in the validation dataset.Conclusions The established prediction model in this study can help in prediction of clinical pregnancy rates following fresh embryo transfer in patients with endometriosis.

Immunotherapy has changed the treatment landscape of advanced non-small cell lung cancer(NSCLC),showing great potential in the treatment of untreated and relapsed or refractory(R/R)patients.However,numerous issues that need further exploration remain with the wide application of immunotherapy.They include the exploration of biomarkers for efficacy prediction,the optimization of immunotherapy modalities,immune-related adverse effects,and the management of special populations.This review summarizes the progress of the research on immunotherapy for advanced NSCLC and discusses its challenges and future directions.