ObjectiveTo explore the target of Erzhiwan in reducing myocardial injury in ovariectomized mice through non-targeted myocardial metabolomics combined with experimental verification. MethodsOvariectomized mouse model was selected， 40 female C57BL/6 mice were randomly divided into sham operation group， model group， estrogen group（estradiol valerate， 1.3×10^-4 g·kg^-1）， Erzhiwan low and high dose groups（3.12， 9.36 g·kg^-1）， with 8 mice in each group. Each administration group was given the corresponding dose of Erzhiwan by gavage， and the sham operation group and model group were given equal volume of distilled water by gavage for 12 weeks. Echocardiography was used to detect cardiac function， hematoxylin-eosin（HE） staining was used to observe myocardial morphological changes， and enzyme-linked immunosorbent assay（ELISA） was used to detect the levels of estrogen， N-terminal pro-brain natriuretic peptide（NT-proBNP）， hypersensitive troponin T（hs-TnT）， total cholesterol（TC）， triglyceride（TG）， low density lipoprotein cholesterol（LDL-C）， high density lipoprotein cholesterol（HDL-C）， interleukin（IL）-1β， IL-18 and tumor necrosis factor-α（TNF-α）. The non-targeted metabolomics of mouse myocardium were analyzed by ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS）， and the differential metabolites and corresponding metabolic pathways were obtained. The mRNA expression levels of phosphatidylinositol 3-kinase（PI3K） and protein kinase B（Akt） in mouse myocardial tissues were detected by real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）， and the protein expression levels of PI3K， Akt， phosphorylated（p）-Akt were detected by Western blot. ResultsCompared with the sham operation group， the model group showed abnormal cardiac function， increased myocardial fiber space， cardiomyocyte atrophy， sarcoplasmic aggregation， and occasional dissolution or rupture of muscle fiber， the level of estrogen in the serum was decreased， the levels of NT-proBNP， hs-TnT， IL-1β， IL-18， TNF-α， TG， TC and LDL-C were increased， and the level of HDL-C was decreased（P<0.01）. Compared with the model group， Erzhiwan could increase the level of estrogen， improve the abnormal cardiac function， reduce the pathological injury of myocardial tissue， decrease the levels of myocardial injury markers（NT-proBNP， hs-TnT） and inflammatory factors（IL-1β， IL-18， TNF-α）， decrease the levels of TG， TC， LDL-C， and increased the level of HDL-C（P<0.01）. The results of non-targeted myocardial metabolomics showed that 31 of the 162 differential metabolites between the model group and sham operation group were significantly adjusted after administration of Erzhiwan， which were mainly glycerol phospholipid metabolites. Pathway enrichment results showed that Erzhiwan mainly affected glycerophospholipid metabolic pathway， PI3K-Akt pathway， cyclic guanosine monophosphate（cGMP）-protein kinase G（PKG） pathway and other metabolic pathways. Compared with the sham operation group， the levels of phosphatidylcholine（PC， 11 types） and phosphatidylethanolamine（PE， 5 types） in mouse myocardial tissue of the model group were increased（P<0.05， P<0.01）， and the mRNA and protein expressions of PI3K and p-Akt were decreased（P<0.05， P<0.01）. Compared with the model group， the levels of PC（11 types） and PE（5 types） were decreased（P<0.05， P<0.01） in myocardial tissue of Erzhiwan group， the mRNA and protein expressions of PI3K and p-Akt were elevated（P<0.01）. ConclusionErzhiwan can alleviate the pathological injury of myocardium in ovariectomized mice， improve the abnormal cardiac function， improve lipid metabolism disorder， and reduce the levels of myocardial injury markers and inflammatory factors， which involves a number of signaling and metabolic pathways in the heart， among which glycerophospholipid metabolism pathway and PI3K/Akt pathway may have key roles.

N6-methyladenosine(m6A)is the most prevalent modification that regulates gene expression in eukaryotes.It regulates splicing,degradation,stability,and translation of RNA.Numerous studies have demonstrated the close association between m6A methylation and tumor development,highlighting its crucial role in regulating tumor immune response.The m6A modification actively participates in governing immune cell differentiation and maturation as well as modulating anti-tumor immune responses.Within the tumor microenvironment,m6A modification can also impact the recruitment,activation,and polarization of immune cells,thereby either promoting or inhibiting tumor cell proliferation and metastasis.Consequently,it plays a pivotal role in reshaping the tumor immune microenvironment.In recent years,immunotherapy for tumors has been increasingly applied to clinical practice with notable success achieved through approaches such as immune checkpoint inhibitor therapy and adoptive cell immunotherapy.Targeting m6A modifications to interfere with the immune system,such as targeting dysregulated m6A regulators through small molecule inhibitors and inducing immune cell reprogramming,can improve anti-tumor immune response and strengthen immune cells' ability to recognize and kill tumor cells.The m6A modification represents a novel avenue for potential clinical application within tumor immunotherapy.This review provides a comprehensive summary of the regulatory impact of m6A methylation modification on immune cells in the context of cancer,while also delving into novel targets for tumor immunotherapy.

Objective:To explore the diagnostic value of serum cystatin C (CysC) and C1q tumor necrosis factor-related protein 9 (CTRP9) levels for diabetic retinopathy (DR) and diabetic macular edema (DME) in patients with type 2 diabetes.Methods:A cross-sectional study was conducted.A total of 135 patients with type 2 diabetes, aged 45-75 years, who were treated in Gansu Provincial Hospital from April 2021 to April 2022 were included.According to DR grading standard, patients were divided into non-DR (NDR) group, non-proliferative DR (NPDR) group and proliferative DR (PDR) group, with 45 patients in each group.The DR patients were subdivided into DME group (51 cases) and non-DME group (39 cases).A total of 45 healthy subjects were selected as the normal control group.Fasting peripheral venous blood was collected to detect serum glycosylated hemoglobin, fasting blood glucose, triacylglycerol, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, CysC and CTRP9 levels.The expression of CysC and CTRP9 levels among different groups were compared.The independent influencing factors of DR and DME were evaluated by multivariate logistic regression analysis model.The diagnostic value of serum CysC and CTRP9 in DR and DME were evaluated by receiver operating characteristic (ROC) curve.This study adhered to the Declaration of Helsinki and the study protocol was approved by the Ethics Committee of Gansu Provincial Hospital (No.2021-301).All patients were informed about the purpose and methods of the study and signed an informed consent form.Results:Serum CysC levels in normal control group, NDR group, NPDR group and PDR group were 0.74(0.67, 0.83), 1.03(0.85, 1.22), 1.40(0.98, 1.63) and 1.66(1.31, 1.85)mg/L, respectively, showing a gradually increasing trend, and the serum CTRP9 levels were (136.90±14.95), (120.23±16.31), (109.50±14.71) and (90.99±13.88)pg/ml, respectively, showing a gradually decreasing trend, with statistically significant overall comparison differences among groups ( Z=89.430, P<0.001; F=74.242, P<0.001), the comparison within groups was statistically significant (all at P<0.05).Compared with non-DME group, the serum CysC level was significantly increased and serum CTRP9 level was significantly decreased in DME group (both P<0.05).Multivariate logistic regression analysis showed that serum CysC (odds ratio [ OR]=19.742, 95% confidence interval [ CI]: 4.515-86.316, P<0.001) was the independent risk influencing factors for the occurrence of DR, and CTRP9 ( OR=0.937, 95% CI: 0.908-0.966, P<0.001) was a protective factor for the occurrence of DR.Serum CTRP9 level ( OR=0.838, 95% CI: 0.778-0.903, P<0.001) was a protective factor for DME.The ROC curve showed that the area under ROC curve (AUC) for serum CysC and CTRP9 levels alone and in combination for the diagnosis of DR in patients with type 2 diabetes mellitus complicated by DR were 0.798, 0.802 and 0.870, respectively.The cutoff values of serum CysC and CTRP9 levels to obtain the best diagnostic efficacy were 1.34 mg/L and 110.12 pg/ml, respectively.The AUC for serum CysC and CTRP9 level alone and in combination for the diagnosis of DME in DR patients were 0.682, 0.923 and 0.923, respectively.The cutoff value of serum CTRP9 level to obtain optimal diagnostic efficacy was 104.68 pg/ml. Conclusions:The enhanced expression of serum CysC level and reduced expression of serum CTRP9 level are the risk factors for the development of DR in type 2 diabetes patients.The decrease of serum CTRP9 level is one of the risk factors for the development of DME in DR patients.

Objective To evaluate the efficacy of medroxyprogesterone acetate(MA)plus metformin as the primary fertility-sparing treatment for atypical endometrial hyperplasia(AEH)and early-stage grade 1 endometrial adenocarcinoma(G1 EAC)and the recurrence rate after treatment.Methods Sixty patients(aged 20-42 years)with AEH and/or grade 1 EAC limited to the endometrium were enrolled prospectively and randomized into two groups(n=30)to receive oral MA treatment at the daily dose of 160 mg(control)or MA plus oral metformin(850 mg,twice a day)for at least 6 months.The treatment could extend to 12 months until a complete response(CR)was achieved,and follow-up hysteroscopy and curettage were performed every 3 months.For all the patients who achieved CR,endometrial expressions of IGFBP-rP1,p-Akt and p-AMPK were detected immunohistochemically.Results A total of 58 patients completed the treatment.After 9 months of treatment,23(76.7%)patients in the combined treatment group and 20(71.4%)in the control group achieved CR;two patients in the control group achieved CR after converting to the combined treatment.The recurrence rate did not differ significantly between the control group and combined treatment group(30.0%vs 22.7%,P>0.05).Ten(35.7%)patients in the control group experienced significant weight gain of 5.7±6.1 kg,while none of the patients receiving the combined treatment exhibited significant body weight changes.Compared with the control group,the patients receiving the combined treatment showed enhanced endometrial expressions of IGFBP-rP1 and p-AMPK with lowered p-Akt expression.Conclusion Metformin combined with MA may provide an effective option for fertility-sparing treatment of AEH and grade 1 stage IA EAC,and the clinical benefits of metformin for controlling MA-induced weight gain and promoting endometrial expressions of IGFBP-rP1 and p-AMPK while inhibiting p-Akt expression warrants further study.

Objective To evaluate the efficacy of medroxyprogesterone acetate(MA)plus metformin as the primary fertility-sparing treatment for atypical endometrial hyperplasia(AEH)and early-stage grade 1 endometrial adenocarcinoma(G1 EAC)and the recurrence rate after treatment.Methods Sixty patients(aged 20-42 years)with AEH and/or grade 1 EAC limited to the endometrium were enrolled prospectively and randomized into two groups(n=30)to receive oral MA treatment at the daily dose of 160 mg(control)or MA plus oral metformin(850 mg,twice a day)for at least 6 months.The treatment could extend to 12 months until a complete response(CR)was achieved,and follow-up hysteroscopy and curettage were performed every 3 months.For all the patients who achieved CR,endometrial expressions of IGFBP-rP1,p-Akt and p-AMPK were detected immunohistochemically.Results A total of 58 patients completed the treatment.After 9 months of treatment,23(76.7%)patients in the combined treatment group and 20(71.4%)in the control group achieved CR;two patients in the control group achieved CR after converting to the combined treatment.The recurrence rate did not differ significantly between the control group and combined treatment group(30.0%vs 22.7%,P>0.05).Ten(35.7%)patients in the control group experienced significant weight gain of 5.7±6.1 kg,while none of the patients receiving the combined treatment exhibited significant body weight changes.Compared with the control group,the patients receiving the combined treatment showed enhanced endometrial expressions of IGFBP-rP1 and p-AMPK with lowered p-Akt expression.Conclusion Metformin combined with MA may provide an effective option for fertility-sparing treatment of AEH and grade 1 stage IA EAC,and the clinical benefits of metformin for controlling MA-induced weight gain and promoting endometrial expressions of IGFBP-rP1 and p-AMPK while inhibiting p-Akt expression warrants further study.

Temporomandibular joint (TMJ) diseases are common clinical conditions. The number of patients with TMJ diseases is large, and the etiology, epidemiology, disease spectrum, and treatment of the disease remain controversial and unknown. To understand and master the current situation of the occurrence, development and prevention of TMJ diseases, as well as to identify the patterns in etiology, incidence, drug sensitivity, and prognosis is crucial for alleviating patients′suffering.This will facilitate in-depth medical research, effective disease prevention measures, and the formulation of corresponding health policies. Cohort construction and research has an irreplaceable role in precise disease prevention and significant improvement in diagnosis and treatment levels. Large-scale cohort studies are needed to explore the relationship between potential risk factors and outcomes of TMJ diseases, and to observe disease prognoses through long-term follw-ups. The consensus aims to establish a standard conceptual frame work for a cohort study on patients with TMJ disease while providing ideas for cohort data standards to this condition. TMJ disease cohort data consists of both common data standards applicable to all specific disease cohorts as well as disease-specific data standards. Common data were available for each specific disease cohort. By integrating different cohort research resources, standard problems or study variables can be unified. Long-term follow-up can be performed using consistent definitions and criteria across different projects for better core data collection. It is hoped that this consensus will be facilitate the development cohort studies of TMJ diseases.

Objective To investigate the protective effects and underlying mechanisms of sodium butyrate on rats exposed to hypoxia and cold conditions.Methods Fifty-eight male SD rats (aged 7～8 weeks,weighing 240～260 g ) were randomly divided into normoxia normothermia saline control (NNC ) group (n=10),normoxia normothermia sodium butyrate(NNB)group(n=10),hypoxia cold saline control (HCC) group (n=19),and hypoxia cold sodium butyrate (HCB)group (n=19).The intragastric dose of sodium butyrate was 200 mg/kg for the NNB and HCB groups,while the NNC and HCC groups were given normal saline of 5 mL/kg.①After continuous intragastric administration for 7 d,the rats in the HCC and HCB groups were placed in a low-pressure hypoxic chamber to simulate an altitude of 5000 m and exposed to a temperature of 8 ℃ for 7 d.Subsequently,blood samples were collected from the abdominal aorta for blood gas analysis,blood routine test,and detection for serum biochemical indicators.ELISA was used to determine serum inflammatory cytokines and endocrine hormones.The rats in the NNC and NNB groups(n=10)were fed outside the chamber and underwent the same tests in 7 d later to evaluate the protective effects of sodium butyrate on the body.②Core body temperature monitoring was conducted to assess the impact of sodium butyrate on the rmoregulation in rats exposed to hypoxia and cold(n=3).③Hypoxia exercise tolerance of the HCC group and HCB group in a hypoxic chamber (11％O2 )was evaluated for their hypoxia resistance (n=6).Results Compared to the NNC group,the rats in the HCC group exhibited significant decreases in arterial oxygen saturation (SaO2 )and arterial oxygen partial pressure (PaO2 ),serum levels of IL-4,estradiol (E2)and cortisol (F),core temperature,and exercise duration (P<0.05),and had notably increased levels of red blood cell (RBC)count,hemoglobin (HGB),hematocrit (HCT),cardiac troponin (CRDAC-T),uric acid (UA),alanine aminotransferase (ALT),total cholesterol (TC),low-density lipoprotein (LDL),IL-6 and granulocyte-macrophage colony-stimulating factor (GM-CSF)(P<0.05).Compared to the HCC group,the rats in the HCB group exhibited significant increases in SaO2,PaO2,IL-4,E2,F,corticotropin releasing hormone (CRH)and adrenocorticotropic hormone (ACTH)(P<0.05),remarkably longer exercise duration under hypoxic exposure (P<0.05 ),but decreases in RBC count,serum levels of HGB,HCT,CRDAC-T,UA,ALT,TC,LDL,IL-6,GM-CSF and free thyroxine (FT4 ),and core temperature (P<0.05).Conclusion Sodium butyrate exhibits protective effects on rats exposed to hypoxia and cold conditions,and it is helpful in their adaptation to these hypoxia and cold environments.

Objective:To investigate the value of diffusion kurtosis imaging (DKI) combined with quantitative dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) in predicting axillary lymph node metastasis in breast cancer.Methods:A total of 150 cases of breast cancer confirmed by pathology in the Affiliated Hospital of Jining Medical University were retrospectively analyzed. 68 cases had axillary lymph node (ALN) metastasis and 82 cases had no ALN metastasis. All breast lesions were examined by DKI and DCE-MRI before operation. We analyzed clinical case data, routine MRI features, DKI, and DCE-MRI parameters between two groups, including diffusion kurtosis (MK), mean diffusion rate (MD), volume transfer constant (K trans), extravascular volume fraction (Ve), and rate constant (Kep); The receiver operating characteristic (ROC) curve was used to analyze the diagnostic efficacy of quantitative parameters for ALN metastasis of breast cancer. Results:The proportion of lesions with blurred edges in the metastatic group was higher than that in the non ALN metastatic group ( P=0.032); The proportion of uneven and circular enhancement within the ALN metastasis group was relatively high ( P=0.018). The MD value of the ALN transfer group was lower than that of the group without ALN transfer ( P=0.021); The MK value, K trans value, and Kep value were higher than those in the group without ALN metastasis (all P<0.01). The K trans value of DCE-MRI model was the most effective in diagnosing ALN metastasis of breast cancer, and the area under the ROC curve (AUC) was 0.831; The AUC of DCE-MRI model was 0.833, which was higher than that of DKI model (AUC=0.733), and the difference was statistically significant ( Z=2.208; P=0.027). The AUC of DCE-MRI and DKI models were higher than that of conventional MRI models ( Z=3.184, P=0.002; Z=1.917, P=0.046). The sensitivity and accuracy of combined DKI and DCE-MRI models in the diagnosis of ALN metastasis in breast cancer were higher than those of single model. Conclusions:DKI and DCE-MRI models can be used to predict axillary lymph node metastasis in breast cancer. Among them, the K trans value of DCE-MRI model is the most effective in diagnosing axillary lymph node metastasis in breast cancer.

Objective To analyze the uptake capacity of the malignant melanoma cell line A375 towards exosomes derived from insulin-like growth factor binding protein 7 (IGFBP7) transfected human adipose-derived mesenchymal stem cells (ADSCs) and the subsequent effects on apoptosis, providing new insights for gene therapy in malignant melanoma. Methods Primary adipose-derived stem cells were prepared by adherent culture of tissue blocks in vitro and passaged to obtain the second-generation. Cell surface markers were identified by cell climbing immunofluorescence. IGFBP7-GFP overexpression vectors were transfected into ADSCs to create ADSCsIGFBP7, followed by measuring the mRNA level of IGFBP7 by fluorescent quantitative PCR. Exosomes were isolated and extracted, and their ultrastructure was observed under electron microscopy. The protein expressions of CD63 and CD81 proteins were detected using Western blot. ADSCsIGFBP7 exosomes were applied to A375 cells, with the uptake capacity of A375 cells for ADSCsIGFBP7 exosomes being observed under a fluorescence microscope. Apoptosis of A375 cells was detected using flow cytometry. Results Immunofluorescence results showed positive expression of CD44 and negative expression of CD34 in the isolated cells, indicating that the isolated and cultured cells displayed a specific phenotype of ADSCs. IGFBP7 mRNA expression in the transfection group was higher compared to the control group (t=11.50, P<0.001), suggesting successful transfection of IGFBP7-GFP overexpression vector into ADSCs to construct ADSCsIGFBP7 cells. Transmission electron microscopy revealed that the exosomes had distinct heterogeneity, showing the morphology of exosomes of ADSCsIGFBP7 appeared as spherical structures of varying sizes with a darker color in the center and low-density substances at the edges. Western blot results indicated positive expression of exosome marker proteins CD63 and CD81, which confirmed the successful extraction of exosomes. After the exosomes of ADSCsIGFBP7 were co-cultured with A375 cells for 6 hours and the exosomes were labeled with PKH26, it was observed that A375 cells exhibited an increased uptake of ADSCIGFBP7 exosomes after 6 hours compared with the control group. The apoptosis rate of A375 cells in the A375 cell group and A375+ADSCIGFBP7 exosome group were (0.743±0.050)% and (4.990±0.196)%, respectively. The difference was statistically significant compared to the control group (t=20.93, P<0.001), with the apoptosis rate of A375+ADSCs IGFBP7 exosome cells being higher. Conclusions ADSCsIGFBP7 exosomes can be taken up by A375 cells and promote their apoptosis.

Ulcerative colitis （UC） is one of the chronic refractory inflammatory bowel diseases characterized by abdominal pain， diarrhea， and mucus， pus and blood in the stool. In recent years， with changes in human life style and improvements of the diagnosis， the incidence and prevalence of UC have been increasing. The pathogenesis of UC is closely related to intestinal mucosal immune dysfunction， intestinal flora disturbance， and abnormal bile acid secretion. Patients with UC have abnormal bile acid secretion and intestinal flora imbalance. A large number of studies have found that abnormal bile acid secretion inhibits immune function， affects signal transduction， and destroys the intestinal mucosal barrier. Intestinal flora disturbance has an important impact on the occurrence and development of inflammation， immune homeostasis， and stress. Bile acids indirectly or directly affect the structure and function of intestinal flora， and at the same time， they produce secondary bile acids under the modification of intestinal flora， entering the liver through enterohepatic circulation. Therefore， the complex dialogue mechanism of bile acid-intestinal flora axis is closely related to the occurrence and development of UC. Based on the basic theory of traditional Chinese medicine（TCM） and clinical research， it is found that emotion is an important factor that induces this disease， spleen and stomach weakness is the root of the disease， and liver depression and spleen deficiency are the key pathogenesis of UC. Combined with modern medicine and molecular biology research， it is believed that abnormal secretion of bile acids is a microscopic manifestation of liver depression in TCM， and intestinal flora disturbance is the biological basis of spleen deficiency. In the pathogenesis of UC， the imbalanced bile acid-intestinal flora axis is consistent with the pathogenesis of liver depression and spleen in TCM. The exploration of the biological connotation of the pathogenesis of UC with liver depression and spleen deficiency from the perspective of bile acid-intestinal flora axis can better explain the scientific nature of its pathogenesis， which provides new clinical solutions and reliable references for studying the pathogenesis of UC with liver depression and spleen deficiency and finding representative prescriptions to prevent and treat this disease.