ObjectiveTo investigate how Xiaoyao Shukun decoction （XYSKD） regulates the formation and release of neutrophil extracellular traps （NETs） via the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway， thereby reducing inflammation， inhibiting the excessive proliferation of fibroblasts in pelvic adhesion tissue， decreasing adhesion and fibrosis， and repairing the tissue damage in sequelae of pelvic inflammatory disease （SPID）. MethodsA total of 84 Wistar rats were randomly allocated into seven groups： blank， model， XYSKD （8 mg·kg^-1）， mTOR agonist （10 mg·kg^-1）， mTOR agonist + XYSKD （10 mg·kg^-1+8 mg·kg^-1）， mTOR inhibitor （2 mg·kg^-1）， and mTOR inhibitor + XYSKD （2 mg·kg^-1+8 mg·kg^-1）. The rat model of SPID was constructed by starvation， fatigue， and ascending Escherichia coli infection. After 14 days of drug intervention， the ultrastructure of fibroblasts in the pelvic adhesion tissue was observed by transmission electron microscopy. The general morphology of the uterus， fallopian tube， and ovary was observed by laparotomy. The levels of interleukin-1β （IL-1β）， interleukin-17 （IL-17）， and tumor necrosis factor-α （TNF-α） in the peritoneal flushing fluid were determined by enzyme-linked immunosorbent assay （ELISA）. The expression of myeloperoxidase （MPO） and citrullinated histone 3 （H3） in the fallopian tube was detected by immunofluorescence. Western blot and Real-time quantitative polymerase chain reaction （Real-time PCR） were employed to determine the relative protein and mRNA levels， respectively， of neutrophil elastase （NE）， intercellular adhesion molecule-1 （CD54）， α-smooth muscle actin （α-SMA）， H3， PI3K， and Akt. ResultsCompared with the blank group， the model group presented a large number of collagen fibers in bundles， numerous cytoplasmic folds of fibroblasts， reduced or absent mitochondrial cristae， and disordered and expanded endoplasmic reticulum. By laparotomy， extensive pelvic congestion， connective tissue hyperplasia， thickening and hardening of the tubal end near the uterus， and tubal and ovarian adhesion or cyst were observed in the model group. In addition， the model group showed raised levels of IL-1β， IL-17， and TNF-α in the peritoneal flushing fluid （P<0.01）， increased average fluorescence intensities of MPO and H3 （P<0.01）， and up-regulated protein and mRNA levels of NE， H3， CD54， PI3K， and Akt （P<0.01）. Compared with the model group， the mTOR agonist group showed increased fibroblasts and cytoplasmic folds， absence of mitochondrial cristae， endoplasmic reticulum dilation， and evident collagen fiber hyperplasia. Pelvic adhesions were observed to cause aggravated damage to the uterine， fallopian tube， and ovarian tissues. The levels of IL-1β， IL-17， and TNF-α in the peritoneal lavage fluid elevated （P<0.01） and the average fluorescence intensities of MPO and H3 enhanced （P<0.01） in the mTOR agonist group. In contrast， the XYSKD group and the mTOR inhibitor group showcased decreased fibroblasts and collagen fibers， alleviated mitochondrial crista loss and endoplasmic reticulum dilation， improved morphology and appearance of the uterine， fallopian tube， and ovarian tissues， lowered levels of IL-1β， IL-17， and TNF-α in the peritoneal lavage fluid （P<0.01）， decreased average fluorescence intensities of MPO and H3 （P<0.01）， and down-regulated protein and mRNA levels of NE， H3， CD54， PI3K， and Akt （P<0.05）. Compared with the mTOR agonist group， the mTOR agonist + XYSKD group showed alleviated pathological changes in the pelvic tissue， declined levels of IL-1β， IL-17， and TNF-α （P<0.01）， decreased average fluorescence intensities of MPO and H3 （P<0.01）， and down-regulated protein levels of NE， H3， CD54， α-SMA， p-PI3K/PI3K， and p-Akt/Akt （P<0.01） and mRNA levels of NE， H3， CD54， α-SMA， PI3K， and Akt （P<0.01）. Compared with the mTOR inhibitor group， the mTOR inhibitor + XYSKD group demonstrated reduced pathological severity of the pelvic tissue， reduced levels of IL-1β， IL-17， and TNF-α （P<0.01）， decreased average fluorescence intensities of MPO and H3 （P<0.01）， and down-regulated protein and mRNA levels of NE and CD54 （P<0.05）. ConclusionXYSKD can inhibit the excessive formation and release of NETs via PI3K/Akt/mTOR to ameliorate the inflammatory environment and reduce fibrosis and adhesion of the pelvic tissue， thereby playing a role in the treatment of SPID. It may exert the effects by lowering the levels of IL-1β， IL-17， and TNF-α and down-regulating the expression of NE， H3， CD54， α-SMA， PI3K， and Akt in the pelvic adhesion tissue.

Objective:To analyze the value of 18F-FDG PET/CT combined with MRI in the diagnosis and differential diagnosis of ductal carcinoma in situ (DCIS) and early stage invasive ductal carcinoma (IDC). Methods:From September 2019 to December 2023, 12 patients with DCIS (all females; age 36-67 years) and 34 patients with early stage IDC (all females; age 36-73 years) in Qingdao Central Hospital were retrospectively analyzed. The general clinical information, MRI features, and 18F-FDG PET/CT features of patients were analyzed. χ2 test, Fisher exact test, and Mann-Whitney U test were used to analyze the data. The independent predictors of DCIS were analyzed by logistic regression analysis. The value of different indicators in diagnosing DCIS was analyzed using ROC curves analysis, and Delong test was used to assess the differences among AUCs. Results:The differences in tumor metabolic volume (MTV; 18.55(10.90, 76.30) vs 4.00(2.00, 11.45)cm 3) and total lesion glycolysis (TLG; 44.85(25.30, 125.30) vs 9.40(6.68, 22.35)g) of breast lesion, enhancement pattern (non-mass enhancement (NME); 8/12 vs 29.4%(10/34)), lobulation sign (0/12 vs 58.8%(20/34)), and apparent diffusion coefficient (ADC; 1.33 (1.16, 1.63)×10 -3vs 1.08 (0.75, 1.28)×10 -3mm 2/s) between DCIS and early stage IDC groups were statistically significant ( Z values: from -3.91 to -2.56, χ2=5.17, all P<0.05). When differentiating DCIS from early stage IDC, NME (odds ratio ( OR)=36.50, 95% CI: 2.15-618.52, P=0.013), ADC ( OR=7.85, 95% CI: 1.11-55.46, P=0.044), and TLG ( OR=1.06, 95% CI: 1.02-1.11, P=0.007) were independent predictors. The AUC of the three predictors combination was 0.941, which was higher than those of single predictors ( Z values: 2.00-2.80, P values: 0.005-0.046). Conclusion:The combination of 18F-FDG PET/CT and MRI improves the efficacy of differential diagnosis between DCIS and early stage IDC, thereby providing a basis for developing personalized treatment plans for patients.

OBJECTIVES: To investigate the effects of early life intervention with Bifidobacterium bifidum BD-1 (B. bifidum BD-1) on hyperactivity in a female mouse model of attention deficit hyperactivity disorder (ADHD) and explore the underlying mechanisms. METHODS: Eight newborn female Wistar-Kyoto (WKY) rats and 6 spontaneous hypertensive rats (SHRs) were gavaged with saline and another 6 SHRs were gavaged with B. bifidum BD-1 (10⁹ CFU) daily for 3 weeks. Open field test of the rats was conducted at 7 weeks, and fecal samples were collected at weaning (3 weeks) and at 7 weeks for 16S rRNA sequencing. Immunofluorescent staining was used to detect dopamine transporter (DAT) and tyrosine hydroxylase (Th) levels in the striatum and activated microglia in the prefrontal cortex. Treg cells in the mesenteric lymph nodes, spleen and blood were analyzed using flow cytometry. RESULTS: The SHRs traveled a significantly greater distance in open fields test than WKY rats, and this behavior was significantly attenuated by B. bifidum BD-1 intervention. The expression of DAT and Th in the striatum was significantly lower in the SHRs than in WKY rats, while B. bifidum BD-1 treatment obviously increased Th levels in the SHRs. B. bifidum BD-1 intervention significantly deceased the number of activated microglia and increased Treg cell counts in the spleen of SHRs. The treatment also enhanced α diversity in gut microbiota of the SHRs and resulted in a decreased Firmicutes/Bacteroidota ratio, more active Muribaculaceae growth, and suppression of Clostridia_UCG-014 proliferation. CONCLUSIONS Early life intervention with B. bifidum BD-1 alleviates hyperactivity in female SHRs by modulating the gut microbiota and peripheral immune response, suppressing neuroinflammation and improving dopaminergic system function. These findings provide evidence for early prevention strategies and support the development and application of psychobiotics for ADHD.
Animals ; Female ; Rats ; Rats, Inbred WKY ; Rats, Inbred SHR ; Attention Deficit Disorder with Hyperactivity/therapy* ; Bifidobacterium bifidum ; Probiotics/therapeutic use* ; Dopamine Plasma Membrane Transport Proteins/metabolism* ; Tyrosine 3-Monooxygenase/metabolism* ; Gastrointestinal Microbiome ; Disease Models, Animal

Objective To explore the diagnostic value of 18F-fluorodeoxyglucose(FDG)PET/CT in primary pulmonary enteric adenocarcinoma.Methods The clinical and imaging data of 9 patients with primary pulmonary enteric adenocarcinoma who under-went 18F-FDG PET/CT examination were retrospectively analyzed,including lesion distribution,morphology,maximum standardized uptake value(SUVmax),clinical symptoms and signs,gastroscopy finding,puncture pathological results,and serum tumor markers[carbohydrate antigen 72-4(CA72-4),cytokeratin 19 fragment antigen 21-1(CYFRA21-1),carcinoembryonic antigen(CEA),carbo-hydrate antigen 199(CA199)].Results Pathological examination confirmed a diagnosis of primary pulmonary enteric adenocarcinoma after excluding gastrointestinal primary tumors through clinical evaluation.In all nine patients,18F-FDG PET/CT examination did not reveal any evidence of digestive system malignancies,and gastrointestinal microscopy was negative.Primary lesions were observed as masses or nodular types in 6 cases(5 in the left lung and 1 in the right lung),while 3 cases exhibited diffuse bilateral pulmonary involvement(manifested as multiple patchy opacities,nodules,ground-glass opacities,and consolidations).All pulmonary primary lesions showed increased 18F-FDG uptake,with SUVmax ranging from 2.7 to 12.8,mean 8.6±3.7.The six masses-or nodular-type primary lesions showed maximum diameters ranging from 2.1 to 10.5 cm,mean(5.23±3.06)cm.Four cases demonstrated hilar and mediastinal lymph node metastases,intrapulmonary metastases,and distant metastases,while 1 case showed only distant metastasis.Elevated levels of serum tumor markers were observed as follows:CA72-4 in 7 cases(10-273.3 U/mL),CEA in 7 cases(5-147.4 ng/mL),CA199 in 6 cases(31.22-4 364 U/mL),and CYFRA21-1 in 5 cases(8.31-99.7 ng/mL).Conclusion When pathological biopsy of a pulmonary lesion suggests primary pulmonary enteric adenocarcinoma after excluding gastrointestinal primary tumors,and 18F-FDG PET/CT shows no gastrointestinal masses,this may support the diagnosis of primary pulmonary enteric adenocarcinoma.

Objective To explore the diagnostic value of 18F-fluorodeoxyglucose(FDG)PET/CT in primary pulmonary enteric adenocarcinoma.Methods The clinical and imaging data of 9 patients with primary pulmonary enteric adenocarcinoma who under-went 18F-FDG PET/CT examination were retrospectively analyzed,including lesion distribution,morphology,maximum standardized uptake value(SUVmax),clinical symptoms and signs,gastroscopy finding,puncture pathological results,and serum tumor markers[carbohydrate antigen 72-4(CA72-4),cytokeratin 19 fragment antigen 21-1(CYFRA21-1),carcinoembryonic antigen(CEA),carbo-hydrate antigen 199(CA199)].Results Pathological examination confirmed a diagnosis of primary pulmonary enteric adenocarcinoma after excluding gastrointestinal primary tumors through clinical evaluation.In all nine patients,18F-FDG PET/CT examination did not reveal any evidence of digestive system malignancies,and gastrointestinal microscopy was negative.Primary lesions were observed as masses or nodular types in 6 cases(5 in the left lung and 1 in the right lung),while 3 cases exhibited diffuse bilateral pulmonary involvement(manifested as multiple patchy opacities,nodules,ground-glass opacities,and consolidations).All pulmonary primary lesions showed increased 18F-FDG uptake,with SUVmax ranging from 2.7 to 12.8,mean 8.6±3.7.The six masses-or nodular-type primary lesions showed maximum diameters ranging from 2.1 to 10.5 cm,mean(5.23±3.06)cm.Four cases demonstrated hilar and mediastinal lymph node metastases,intrapulmonary metastases,and distant metastases,while 1 case showed only distant metastasis.Elevated levels of serum tumor markers were observed as follows:CA72-4 in 7 cases(10-273.3 U/mL),CEA in 7 cases(5-147.4 ng/mL),CA199 in 6 cases(31.22-4 364 U/mL),and CYFRA21-1 in 5 cases(8.31-99.7 ng/mL).Conclusion When pathological biopsy of a pulmonary lesion suggests primary pulmonary enteric adenocarcinoma after excluding gastrointestinal primary tumors,and 18F-FDG PET/CT shows no gastrointestinal masses,this may support the diagnosis of primary pulmonary enteric adenocarcinoma.

Objective:To analyze the value of 18F-FDG PET/CT combined with MRI in the diagnosis and differential diagnosis of ductal carcinoma in situ (DCIS) and early stage invasive ductal carcinoma (IDC). Methods:From September 2019 to December 2023, 12 patients with DCIS (all females; age 36-67 years) and 34 patients with early stage IDC (all females; age 36-73 years) in Qingdao Central Hospital were retrospectively analyzed. The general clinical information, MRI features, and 18F-FDG PET/CT features of patients were analyzed. χ2 test, Fisher exact test, and Mann-Whitney U test were used to analyze the data. The independent predictors of DCIS were analyzed by logistic regression analysis. The value of different indicators in diagnosing DCIS was analyzed using ROC curves analysis, and Delong test was used to assess the differences among AUCs. Results:The differences in tumor metabolic volume (MTV; 18.55(10.90, 76.30) vs 4.00(2.00, 11.45)cm 3) and total lesion glycolysis (TLG; 44.85(25.30, 125.30) vs 9.40(6.68, 22.35)g) of breast lesion, enhancement pattern (non-mass enhancement (NME); 8/12 vs 29.4%(10/34)), lobulation sign (0/12 vs 58.8%(20/34)), and apparent diffusion coefficient (ADC; 1.33 (1.16, 1.63)×10 -3vs 1.08 (0.75, 1.28)×10 -3mm 2/s) between DCIS and early stage IDC groups were statistically significant ( Z values: from -3.91 to -2.56, χ2=5.17, all P<0.05). When differentiating DCIS from early stage IDC, NME (odds ratio ( OR)=36.50, 95% CI: 2.15-618.52, P=0.013), ADC ( OR=7.85, 95% CI: 1.11-55.46, P=0.044), and TLG ( OR=1.06, 95% CI: 1.02-1.11, P=0.007) were independent predictors. The AUC of the three predictors combination was 0.941, which was higher than those of single predictors ( Z values: 2.00-2.80, P values: 0.005-0.046). Conclusion:The combination of 18F-FDG PET/CT and MRI improves the efficacy of differential diagnosis between DCIS and early stage IDC, thereby providing a basis for developing personalized treatment plans for patients.

Objective To investigate the imaging features and pathogenesis of lethal iatrogenic hemobilia(LIH)and the value of transarterial intervention in the treatment of LIH.Methods A total of 269 patients with upper gastrointestinal bleeding who were admitted to The First Affiliated Hospital of Nanjing Medical University from August 2009 to July 2023 were enrolled,among whom 24 had a confirmed diagnosis of LIH and received treatment,and a retrospective analysis was performed for the clinical data of these 24 patients,including the iatrogenic causes,angiographic findings,and arterial interventions of LIH.Among the 24 patients,23 received transarterial embolization(TAE)with gelatin sponge particles and coils,and 1 received a covered stent for isolation.The main criteria for assessing treatment outcome included the technical success rate of surgery,procedure-related complications,and long-term clinical follow-up.Results Among the 24 patients with LIH,12 had LIH caused by interventional procedures,and 12 had LIH caused by hepatobiliary and pancreatic surgery.The main clinical manifestations included a significant reduction in blood pressure or a persistent reduction in hemoglobin in 13 patients and upper gastrointestinal bleeding in 18 patients.Among the 24 patients,2 developed symptoms during surgery,4 developed symptoms within 24 hours,and 18 developed symptoms after 24 hours.Angiography showed a positive bleeding rate of 100%(24/24),and imaging findings included pseudoaneurysms in 15 patients,hepatic artery truncation in 3 patients,extravasation of contrast medium in 5 patients,and hepatic arteriobiliary fistula in 3 patients.Among the 24 patients,23 received TAE and 1 received stent implantation.Successful hemostasis was achieved for 23 patients,with a technical success rate of 95.8%(23/24).Four patients developed hepatic necrosis and abscess after TAE,and there was no rebleeding or recurrence after hemostatic treatment.Conclusion Various iatrogenic injuries may result in LIH with diverse clinical and imaging findings,and integrated diagnostic imaging combined with transarterial intervention is the best effective life-saving measure for LIH.

Objective To study the changes in peripheral blood T lymphocytes in patients with ankylosing spondylitis and their correlation with the disease's severity and prognosis.Methods We selected 120 patients with ankylosing spondylitis treated between January 2020 and March 2023 as the research group and 120 healthy people who had medical examinations in the same period as the health group.We detected the changes in CD4+,CD8+and CD4+/CD8+values of peripheral blood T lymphocyte subsets with flow cytometry,compared the differences in T lymphocyte subsets between the two groups,and analyzed the correlation with the disease severity of the patients.All the 120 patients with ankylosing spondylitis were followed up for 6 months after treatment to assess their prognosis.General information and T lymphocyte sub-groups CD4+,CD8+level,CD4+/CD8+value changes were compared among patients with different prognosis.We analyzed the value of T lymphocyte sub-groups in predicting the prognosis of patients with ankylosing spondylitis.Results In the research group CD4+and CD4+/CD8+were lower but CD8 1 was higher than those in the healthy group(P＜0.05).CD4+and CD4+/CD8 were lower but CD8+was higher in patients with advanced ankylosing spondylitis than in early and mid-term patients(P＜0.05).The ROC curve analysis showed that the AUC of CD4+,CD8+,and CD4+/CD8+combined diagnosis of ankylosing spondylitis patients was 0.878,with higher diagnostic sensitivity than that of the single diagnosis(P＜0.05).In the poor prognosis group,CD8+was higher than that in the excellent prognosis group,but CD4+and CD4+/CD8 value were lower than the latter(P＜0.05).The results of Pearson test showed that CD4+and CD4+/CD8+were negatively correlated with the prognosis of patients with ankylosing spondylitis(r=-0.568,-0.656,P＜0.001).CD8+was positively correlated with the prognosis of patients with ankylosing spondylitis(r=0.623,P＜0.001).ROC curve analysis showed that the AUC of the combined diagnosis of CD4+,CD8+and CD4+/CD8+for ankylosing spondylitis patients was 0.910,and the diagnostic sensitivity was higher than that of single diagnosis(P＜0.05).Conclusion The abnormal levels of peripheral blood T lymphocyte subsets in patients with ankylosing spondylitis are closely related to the severity and prognosis of the disease,and can be used as a reference indicator for diagnosing the severity and prognosis of ankylosing spondylitis.

Objective We aimed to explore the mechanism by which Bushen Yangjing Soup improves elderly infertility based on oocyte mitochondrion.Methods C57BL/6J mice were used,including 15 young females(6-8 weeks old),45 elderly females(10 months old),and ten young males(6-8 weeks old).The female 6-8-week-old young mice were assigned to the young group,and the female 10-month-old elderly mice were randomly divided into the model group,the dehydroepiandrosterone(DHEA)group,and the Bushen Yangjing Soup group.Mice in the DHEA and Bushen Yangjing Soup groups were given DHEA[11.26 mg/(kg·d)]and Bushen Yangjing Soup[11.41 g/(kg·d)].Mice in the other groups were given normal saline.After 45 days,oocytes were collected,the number of oocytes and the proportion of mature oocytes were recorded,and in vitro fertilization(IVF)was carried out,followed by calculation of the successful fertilization rate.Real-time PCR was used to determine the copy number of mitochondrial DNA in oocytes,mito-tracker green was used to observe the distribution of mitochondria,and the mitochondrial membrane potential(MMP)of oocytes was determined using the JC-1 kit.The mRNA and protein levels of mitofusin 2(Mfn2)in oocytes were determined by real-time PCR and Western blotting,respectively.Results Compared with the young group,the number of oocytes,the proportion of mature oocytes,and the IVF rate were significantly reduced(P＜0.01),the copy number of mitochondrial DNA in oocytes was decreased,the rate of abnormal mitochondrial distribution was increased,the MMP was decreased(P＜0.01),and the mRNA and protein levels of Mfn2 were decreased(P＜0.01)in the elderly model mice.Compared with the elderly model group,the number of oocytes,the proportion of mature oocytes,and the IVF rate were significantly increased(P＜0.05),the copy number of mitochondrial DNA in oocytes was increased(P＜0.05),the rate of abnormal mitochondrial distribution was decreased,the MMP was increased(P＜0.05),and the mRNA and protein levels of Mfn2 were increased(P＜0.01)in the Bushen Yangjing Soup group.Compared with the DHEA group,the number of oocytes,the proportion of mature oocytes,and the IVF rate were increased,the copy number of mitochondrial DNA in oocytes was increased,the rate of abnormal mitochondrial distribution was decreased,the MMP was increased,the mRNA and protein levels of Mfn2 were increased in the Bushen Yangjing Soup group,and there was a significant difference in the number of oocytes and the expression of Mfn2 protein(P＜0.05).Conclusion Bushen Yangjing Soup may improve the quantity,distribution,and function of oocyte mitochondria by increasing the mRNA and protein levels of Mfn2,thereby improving oocyte quality.This is one of the possible mechanisms for clinical application of Bushen Yangjing Soup to improve ovarian function and treat elderly infertility.

Background::Hyperglycemia frequently induces apoptosis in endothelial cells and ultimately contributes to microvascular dysfunction in patients with diabetes mellitus (DM). Previous research reported that the expression of integrins as well as their ligands was elevated in the diseased vessels of DM patients. However, the association between integrins and hyperglycemia-induced cell death is still unclear. This research was designed to investigate the role played by integrin subunit β5 (ITGB5) in hyperglycemia-induced endothelial cell apoptosis.Methods::We used leptin receptor knockout (Lepr-KO) ( db/ db) mice as spontaneous diabetes animal model. Selective deletion of ITGB5 in endothelial cell was achieved by injecting vascular targeted adeno-associated virus via tail vein. Besides, we also applied small interfering RNA in vitro to study the mechanism of ITGB5 in regulating high glucose-induced cell apoptosis. Results::ITGB5 and its ligand, fibronectin, were both upregulated after exposure to high glucose in vivo and in vitro. ITGB5 knockdown alleviated hyperglycemia-induced vascular endothelial cell apoptosis and microvascular rarefaction in vivo. In vitro analysis revealed that knockdown of either ITGB5 or fibronectin ameliorated high glucose-induced apoptosis in human umbilical vascular endothelial cells (HUVECs). In addition, knockdown of ITGB5 inhibited fibronectin-induced HUVEC apoptosis, which indicated that the fibronectin-ITGB5 interaction participated in high glucose-induced endothelial cell apoptosis. By using RNA-sequencing technology and bioinformatic analysis, we identified Forkhead Box Protein O1 (FoxO1) as an important downstream target regulated by ITGB5. Moreover, we demonstrated that the excessive macroautophagy induced by high glucose can contribute to HUVEC apoptosis, which was regulated by the ITGB5-FoxO1 axis. Conclusion::The study revealed that high glucose-induced endothelial cell apoptosis was positively regulated by ITGB5, which suggested that ITGB5 could potentially be used to predict and treat DM-related vascular complications.