This study explored the potential therapeutic targets and mechanisms of Danggui Shaoyao San(DSS) in the prevention and treatment of Alzheimer's disease(AD) through transcriptomics and metabolomics, combined with animal experiments. Fifty male C57BL/6J mice, aged seven weeks, were randomly divided into the following five groups: control, model, positive drug, low-dose DSS, and high-dose DSS groups. After the intervention, the Morris water maze was used to assess learning and memory abilities of mice, and Nissl staining and hematoxylin-eosin(HE) staining were performed to observe pathological changes in the hippocampal tissue. Transcriptomics and metabolomics were employed to sequence brain tissue and identify differential metabolites, analyzing key genes and metabolites related to disease progression. Reverse transcription-quantitative polymerase chain reaction(RT-qPCR) was employed to validate the expression of key genes. The Morris water maze results indicated that DSS significantly improved learning and cognitive function in scopolamine(SCOP)-induced model mice, with the high-dose DSS group showing the best results. Pathological staining showed that DSS effectively reduced hippocampal neuronal damage, increased Nissl body numbers, and reduced nuclear pyknosis and neuronal loss. Transcriptomics identified seven key genes, including neurexin 1(Nrxn1) and sodium voltage-gated channel α subunit 1(Scn1a), and metabolomics revealed 113 differential metabolites, all of which were closely associated with synaptic function, oxidative stress, and metabolic regulation. RT-qPCR experiments confirmed that the expression of these seven key genes was consistent with the transcriptomics results. This study suggests that DSS significantly improves learning and memory in SCOP model mice and alleviates hippocampal neuronal pathological damage. The mechanisms likely involve the modulation of synaptic function, reduction of oxidative stress, and metabolic balance, with these seven key genes serving as important targets for DSS in the treatment of AD.
Animals ; Alzheimer Disease/genetics* ; Male ; Drugs, Chinese Herbal/administration & dosage* ; Mice ; Mice, Inbred C57BL ; Metabolomics ; Transcriptome/drug effects* ; Maze Learning/drug effects* ; Hippocampus/metabolism* ; Humans ; Disease Models, Animal ; Memory/drug effects*

Objective : To explore the impacts and fundamental mechanisms of the PU. 1 inhibitor DB2313 on the immune function in MRL/lpr mice. Methods: A total of thirty MRL/lpr lupus mice were randomly distributed into three separate groups : the model control group , the PU. 1 inhibitor DB2313 treatment group ( administered at a dose of 17 mg/kg) , and the positive drug control Telitacicept (TACI_Ig) group (administered at a dose of 7. 5 mg/ kg) . Furthermore , a group of ten BALB/c mice were assigned as the normal group. The DB2313 administration group was treated with intraperitoneal injections of the drug on three occasions per week , while the TACI_Ig group received subcutaneous injections every second day; both treatment protocols were maintained for a duration of five weeks. Both the control group and the model group were administered intraperitoneal injections of a volume of saline that was equivalent across groups. After the drug was given , mice were sacrificed by dislocation after orbital vein blood collection. The thymus and spleen were aseptically excised , individually weighed , and subsequently utilized to compute the thymus index and spleen index. The relative distribution of T lymphocyte subsets within the spleen was ascertained through flow cytometry. Serum concentrations of anti_nuclear antibodies ( ANA) and antidouble_stranded DNA antibodies were quantified using an enzyme_linked immunosorbent assay (ELISA) . The levels of inflammatory factors interleukin_6 (IL_6) , tumor necrosis factor_α (TNF_α) , interferon_γ(IFN_γ) were meas ured by CBA method. Hematoxylin and eosin ( HE) staining was employed to examine pathological alterations in the spleen. The expression of PU. 1 and IL_9 in spleen tissue was detected using immunohistochemistry. Additionally , the expression level of PU. 1 protein in the spleen tissue was ascertained through Western blot analysis. Results: The administration of DB2313 significantly ameliorated spleen lesions in MRL/lpr mice and decreased the levels of anti_ds_DNA , ANA , TNF_α , IL_6 , and IFN_γ . It also reduced the proportion of total T cells , TFH cells , Th17 cells , and Th9 cells in the mouse spleen , while increasing the proportion of Treg cells. Furthermore , it lowered the level of PU. 1 protein in the spleen. Immunohistochemistry results demonstrated that DB2313 treatment significantly diminished the expression of PU. 1 and IL_9 in spleen tissue. Conclusion The PU. 1 inhibitor DB2313 can improve spleen lesions in MRL/lpr mice and slow the progression of the disease , and its mechanism is related to the regulation of immune cell functions.

Objective : To establish an interleukin-1β (Il-1β) induced inflammatory model of rat articular chondro- cytes (ACs) , and to investigate the relationship between the expression of lysine acetyltransferase 7 (KAT7) under inflammatory stimulation and the senescence of ACs. Methods: Primary ACs were obtained by digestion of rat knee cartilage with collagenase type Ⅱ and identified. The inflammatory model of ACs was induced by IL-1β . KAT7 was over-expressed or knocked down in ACs by adeno-associated virus infection or small interfering RNA transfection , respectively. A negative control group was set up. Transwell assay was used to detect cell migration ability. Senes- cent cells were stained with senescence-associated β-galactosidase (SA-β-Gal) . Western blot ( WB) was used to detect the protein expression levels of KAT7 , collagen type II (Col Ⅱ ) , matrix metalloproteinase 13 (MMP13) , tumor protein p53 (p53) and cyclin-dependent kinase inhibitor 1A (p21) . The cells of negative control group and KAT7 over-expression group were performed for RNA sequencing , and WB was used to verify the related signaling pathways obtained by Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis. Results: Compared with the control group , the SA-β-Gal staining was enhanced , the protein expression of Col Ⅱ decreased , the pro- tein expression of MMP13 and p53 increased , the cell migration ability decreased , and the expression of KAT7 also increased in the ACs of rats after IL-1β stimulation. Compared with the negative control group , the SA-β-Gal stai- ning was enhanced , the protein expression of Col Ⅱ decreased , the protein expression of MMP13 , p53 and p21 in- creased , and the cell migration ability decreased in the KAT7 over-expression group. Compared with the negative control group , the SA-β-Gal staining was weakened , the protein expression of Col Ⅱ increased , the protein expres- sion of MMP13 , p53 and p21 decreased , and the cell migration ability was enhanced in the KAT7 knockdown inflammatory model of ACs. KEGG enrichment analysis showed that phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway was activated. Compared with the negative control group , the relative protein ex⁃pression levels of phosphorylated protein kinase B (p⁃AKT)/AKT and phosphorylated mammalian target of rapamy⁃cin (p⁃mTOR)/mTOR in KAT7 over⁃expression group increased. The relative protein expression levels of p ⁃AKT/AKT and p ⁃mTOR/mTOR in KAT7 knockdown cells decreased. Conclusion Rat ACs with high expression of KAT7 exhibit senescence and osteoarthritis phenotype , and the mechanism may be related to the activation of PI3K/AKT/mTOR signaling pathway by KAT7.

Objective To explore the impacts and fundamental mechanisms of the PU.1 inhibitor DB2313 on the immune function in MRL/lpr mice.Methods A total of thirty MRL/lpr lupus mice were randomly distributed into three separate groups:the model control group,the PU.1 inhibitor DB2313 treatment group(administered at a dose of 17 mg/kg),and the positive drug control Telitacicept(TACI-Ig)group(administered at a dose of 7.5 mg/kg).Furthermore,a group of ten BALB/c mice were assigned as the normal group.The DB2313 administration group was treated with intraperitoneal injections of the drug on three occasions per week,while the TACI-Ig group received subcutaneous injections every second day;both treatment protocols were maintained for a duration of five weeks.Both the control group and the model group were administered intraperitoneal injections of a volume of sa-line that was equivalent across groups.After the drug was given,mice were sacrificed by dislocation after orbital vein blood collection.The thymus and spleen were aseptically excised,individually weighed,and subsequently uti-lized to compute the thymus index and spleen index.The relative distribution of T lymphocyte subsets within the spleen was ascertained through flow cytometry.Serum concentrations of anti-nuclear antibodies(ANA)and anti-double-stranded DNA antibodies were quantified using an enzyme-linked immunosorbent assay(ELISA).The lev-els of inflammatory factors interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ)were meas-ured by CBA method.Hematoxylin and eosin(HE)staining was employed to examine pathological alterations in the spleen.The expression of PU.1 and IL-9 in spleen tissue was detected using immunohistochemistry.Addition-ally,the expression level of PU.1 protein in the spleen tissue was ascertained through Western blot analysis.Re-sults The administration of DB2313 significantly ameliorated spleen lesions in MRL/lpr mice and decreased the levels of anti-ds-DNA,ANA,TNF-α,IL-6,and IFN-γ.It also reduced the proportion of total T cells,TFH cells,Th17 cells,and Th9 cells in the mouse spleen,while increasing the proportion of Treg cells.Furthermore,it low-ered the level of PU.1 protein in the spleen.Immunohistochemistry results demonstrated that DB2313 treatment sig-nificantly diminished the expression of PU.1 and IL-9 in spleen tissue.Conclusion The PU.1 inhibitor DB2313 can improve spleen lesions in MRL/lpr mice and slow the progression of the disease,and its mechanism is related to the regulation of immune cell functions.

Objective To investigate the impact of corona virus disease 2019(COVID-19)vaccination on incidence of COVID-19 pneumonia and pulmonary function in early convalescence phase.Methods A total of 1 178 COVID-19 patients from Zhongshan hospital,Fudan University were enrolled from 7 Jan 2023 to 7 Feb 2023 and divided into two groups according to the incidence of COVID-19 pneumonia.All the patients were asked to finish the questionnaire on COVID-19 related-symptoms,and other clinical parameters in our study.Vaccination situation,Pulmonary functions including ventilation,diffusion function and fractional exhaled nitricoxide(FeNO)were collected.T-tests and Chi-squared tests were used to analyze the protective effect of vaccination on pulmonary function and occurrence of COVID-19 pneumonia between groups.Univariate analysis and multivariate analysis were performed to predict the significant prognostic factors for incidence of COVID-19 pneumonia.Results Lung function including ventilation and diffusion function was significantly declined in unvaccinated patients than vaccinated patients(P<0.05).Those unvaccinated patients and patients with chronic diseases were more likely to have COVID-19 pneumonia and severe related symptoms.Vaccinated with two or three doses,rather than one dose of COVID-19 vaccines could protect the patients from COVID-19 pneumonia(P<0.05).Those with chronic diseases were inclined to have severe symptoms,while vaccination could alleviate the symptoms of those patients and patients with no history.Conclusion COVID-19 infection can impair the lung function of patients in early convalescence phase.COVID-19 vaccination could protect the patients from COVID-19 pneumonia,ameliorate the related symptoms and attenuate lung function decline.

Aim To investigate the protective poten-tial of paeoniflorin 6-oxy-benzenesulfonate(CP-25)in preserving knee cartilage integrity in osteoarthritis mice through inhibition of GRK2 activity.Methods The posttraumatic osteoarthritis model was established fol-lowing DMM surgery.The experiment consisted of a sham operation group,a model group,a CP-25 admin-istration group,and a paroxetine positive control group.Intragastric administration commenced after the surgery.After 12 weeks of administration,the animals were euthanized.Micro-CT imaging was used to ob-serve the knee cartilage degeneration and abnormal bone remodeling,and the joint histopathology of mice was observed by staining with ferrubens solid green.Immunohistochemistry and immunofluorescence were used to detect the expression level of related molecules in cartilage tissue.Furthermore,Western blot was em-ployed to determine GRK2 and EP4 membrane protein expression levels as well as total protein levels of GRK2 and MMP13 following CP-25 treatment.Results Compared with the sham operation group,the articular cartilage in the model group was significantly degrad-ed,with the cartilage surface calcifying and osteo-phytes forming.CP-25 could significantly reduce the number of osteophytes and the thickness of the sub-chondral plate of articular cartilage,promote the regen-eration of the cartilage matrix,reduce the expression of cartilage matrix degradation protein,and have a signifi-cant protective effect on knee cartilage.Immunohisto-chemical and immunofluorescence results showed that compared with the model group,CP-25 treatment sig-nificantly decreased the expressions of GRK2,AD-AMTS5 and MMP13 in knee tissue and increased the expressions of Col Ⅱ and Aggrecan in knee tissue.The results of in vitro experiments showed that CP-25 ad-ministration could significantly reduce the expression levels of GRK2 membrane protein and total protein,in-crease the level of EP4 membrane protein,and de-crease the level of MMP13.Conclusions The ad-ministration of CP-25 can significantly promote the re-generation of articular cartilage matrix in OA mice,re-duce the degradation of cartilage matrix,and exhibit therapeutic effects on OA.The mechanism behind this is related to the inhibition of GRK2-mediated cartilage matrix metabolism.

OBJECTIVE: To explore the immunological mechanisms underlying spermatogenetic malfunction in patients with non-obstructive azoospermia (NOA) based on bioinformatics and machine learning, and to screen out the key genes associated with spermatogenesis failure. METHODS: NOA-related datasets were obtained from the GEO database, and the differentially expressed genes identified by differential analysis and weighted gene co-expression network analysis (WGCNA). A model of spermatogenesis scoring was established for analysis of the immunological microenvironment and cell interaction networks related to spermatogenesis failure. The key genes were screened out by machine learning, followed by analysis of their correlation with T cells and macrophages. An NOA mouse model was constructed for validation of transcriptome sequencing. RESULTS: Seventy-five differentially expressed genes were identified for the establishment of the spermatogenesis scoring model. The low spermatogenesis score group showed a higher infiltration of the immune cells, with an increased proportion of T cells and macrophages and a correlation of cell interaction signals with immunity. SOX30, KCTD19, ASRGL1 and DRC7 were identified by machine learning as the key genes related to spermatogenesis, with down-regulated expressions in the NOA group, and their expression levels negatively correlated with the infiltration of T cells and macrophages. The accuracy of the spermatogenesis scoring and machine learning models, as well as the trend of the expression levels of the key genes, was successfully validated with the transcriptome sequencing data on the NOA mouse testis. CONCLUSION The development of NOA is closely associated with enhanced immunological microenvironment in the testis. T cells and macrophages may play important roles in spermatogenesis failure. SOX30, KCTD19, ASRGL1 and DRC7 are potential biomarkers for the diagnosis and treatment of NOA.
Male ; Azoospermia/genetics* ; Machine Learning ; Animals ; Computational Biology ; Mice ; Humans ; Spermatogenesis/genetics* ; Gene Expression Profiling ; Macrophages/immunology* ; Gene Regulatory Networks ; T-Lymphocytes/immunology* ; Transcriptome

OBJECTIVE: To investigate the value of pretreatment inflammatory-nutritional biomarkers in predicting the pathological response of locally advanced rectal cancer (LARC) after neoadjuvant chemotherapy (nCT). METHODS: This retrospective study included eligible participants who underwent nCT followed by radical surgery. Pretreatment inflammatory nutritional biomarkers were calculated within one week prior to nCT. Correlations between biomarkers and pathological responses were analyzed. The cut-off values of the pretreatment biomarkers for predicting non-response were determined using receiver operating characteristic (ROC) curve analysis. The inflammation-nutrition score was calculated using the lymphocyte level, neutrophil-to-lymphocyte ratio (NLR), and prognostic nutritional index (PNI). RESULTS: A total of 235 patients were retrospectively recruited between January 2017 and September 2022. Lower lymphocyte levels, lymphocyte monocyte ratio (LMR), and PNI, and higher NLR and platelet-to-lymphocyte ratio (PLR) were observed in patients without response. Multivariate logistic regression analysis revealed that NLR could independently predict non-response to nCT in patients with LARC. The sensitivity and specificity of the inflammation-nutrition score for predicting nonresponse were 71.2% and 61.7%, respectively. CONCLUSION The pretreatment inflammation-nutrition score is a practical parameter for predicting non-response to nCT in patients with LARC. Patients with high scores were more likely to respond poorly to nCT.
Humans ; Retrospective Studies ; Neoadjuvant Therapy ; Lymphocytes ; Biomarkers ; Rectal Neoplasms/pathology*

Objective To compare the prevalence and disease burden of thyroid cancer and their trends between China and the globe from 1990 to 2019.Methods With the global disease burden data in 2019,Joinpoint was used to predict the trends of the disease burden of thyroid cancer in China and the globe from 1990 to 2019,and logarithmic linear model was used to test the predicted trends.The R language was used for predictive analysis and graphic plotting of the disease burden from 2020 to 2035.Results From 1990 to 2019,the standardized incidence rate and the standardized mortality rate of thyroid cancer in China were lower than those in the globe.The standardized incidence rate in China and the globe showed an increasing trend(with the increases of 102.65% and 40.65%,respectively),while the standardized mortality rate showed a decreasing trend(with the decreases of 7.63% and 4.91%,respectively).Compared with those of the female population,the standardized incidence and mortality rates of the Chinese male population increased significantly from 1990 to 2019(the rates of change in the male population were 48.65% and 214.60%,respectively;and the rates of change in the female population were -39.01% and 60.44%,respectively).China's overall standardized years of life lost(YLL),years lived with disability(YLD),and disability-adjusted life years(DALY)rates during the 30-year period were lower than the global average.The Chinese and global populations showed the standardized YLL rate decreasing by 16.61% and 6.88% and the standardized DALY rate decreasing by 10.77% and 3.65%,respectively,while the rates of standardized YLD increased by 128.91% and 46.89%,respectively.The magnitude of DALY in China and the world was mainly influenced by YLL.The standardized incidence,mortality,and DALY rates of the Chinese male population were gradually approaching the global levels.From 1990 and 2019,thyroid cancer showed a higher mortality rate in the population with the age ≥ 75 years and a higher incidence rate in the population with the age <75 years.It is projected that from 2020 to 2035,the standardized incidence rates in China and the world will increase by 36.66% and 21.15%,respectively;the standardized mortality rates will decrease by 20.19% and 3.46%,respectively;and the standardized DALY rate is expected to decrease by 7.08% in China and increase by 4.35% in the world.Conclusions From 1990 to 2019,China's standardized incidence rate of thyroid cancer increased and had a higher increase than the global level,and the standardized mortality rate decreased,with a slightly higher decrease than the global level.However,the increases in the standardized incidence rate and mortality rate of this disease in China's ≥75 years male population were severe.Although China's disease burden of thyroid cancer showed a decreasing trend in line with the global trend as a whole,the disease burden in the Chinese males was higher than that in the females.Specifically,the disease burden due to premature death was predominant,and the burden in specific populations requires policy attention.
Male ; Humans ; Female ; Aged ; Quality-Adjusted Life Years ; Reference Standards ; Cost of Illness ; China/epidemiology* ; Thyroid Neoplasms/epidemiology* ; Incidence

Objective: To investigate the clinical and pathological features, treatments and prognosis of laryngeal neuroendocrine carcinoma (LNEC). Methods: We conducted the retrospective analysis of the clinical data of 12 patients with LNEC admitted to the Department of Otorhinolaryngology Head and Neck Surgery, Second Hospital of Shanxi Medical University from May 2014 to December 2021, including 9 males and 3 females, aged 50-77 years. There were 4 cases of typical carcinoid tumour (highly differentiated), 5 cases of atypical carcinoid tumour (moderately differentiated) and 3 cases of neuroendocrine small cell carcinoma (hypofractionated). The clinical features, diagnosis, treatment and prognosis of LNEC were analysed. Results: The clinical manifestations of LNEC varied according to the tumour type but did not correlate with the pathological types. The supraglottic type was characterized by sore throat, foreign body sensation in the pharynx, coughing, obstructive sensation when eating and choking on water. The treatments were determined according to the pathological types, lesion location and invasion scope. Of 12 patients 4 underwent horizontal partial laryngectomy plus elective lymphatic dissection plus postoperative radiotherapy/chemotherapy, 4 underwent vertical partial laryngectomy (3 of them with cervical lymphatic dissection), 3 underwent supported laryngoscopic plasma laryngectomy for laryngeal cancer, and 1 abandoned for treatment. With the follow-up of 8 -78 months, 5 patients were alive, 1 died from chemotherapy reactions, 3 died from other diseases, 1 died from lung metastasis, 1 died from lung infection and 1 was lost to follow-up. Conclusion: LNEC is clinically rare, the clinical manifestations are less specificity, diagnosis relies on pathological and immunohistochemical examinations, and treatment modalities and prognoses are closely related to the pathological subtypes of LNEC.
Humans ; Male ; Female ; Laryngeal Neoplasms/pathology* ; Retrospective Studies ; Carcinoma, Neuroendocrine/pathology* ; Laryngectomy ; Carcinoid Tumor/pathology*