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.

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 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 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 review the characteristics of animal models of sequelae of pelvic inflammatory disease(SPID)to provide a reference for standardizing the modeling process and improving the modeling rate.Methods Literature relevant to animal models of SPID from the past 40 years was searched,and animal species,modeling method,modeling cycles,modeling substances,positive control drugs,and evaluation indexes were summarized and analyzed.Results A total of 243 study manuscripts were included,most of which induced the SPID model in rats via the phenol paste or microbial infection method.The modeling cycles were typically between 14 and 16 days,and the success of the models was mostly determined by pelvic tissue morphology observation and pathological HE staining.Few research reports have focused on the traditional Chinese medicine(TCM)disease combination model.Conclusions No consistent criteria have been established for SPID animal model modeling,and thus it is recommended that researchers evaluate changes to animal behavior,pelvic histomorphology,and pathology.TCM syndrome modeling lacks effective method and evaluation standards,which need further research and development.Finally,the selection and use of positive-control drugs need to be further explored and perfected.

Objective To study the effect of Honokiol(HKL)on pulmonary microvascular endothelial cells in lipopolysaccharide(LPS)-induced acute respiratory distress syndrome(ARDS)and its potential mechanism.Methods Mouse lung microvascular endothelial cells(PMVECs)were cultured with DMEM+10％FBS in a six-well plate and divided into control(Con)group 1,Honokiol(HKL)group 1,LPS treated(LPS)group 1,and LPS+HKL treatment(HKL+LPS)group 1.The levels of malondialdehyde(MDA)and reactive oxygen species(ROS)in cell lysates were determined by lipid peroxidation assay kit and H2DCF-DA,respectively.TUNEL/DAPI double staining was used to detect apoptosis.Cell junctions were visualized via VE-cadherin/DAPI and Claudin-5/DAPI double staining.Western blot was used to detect caspase-3,cleaved caspase-3,Sirt3,SOD2,and acetylated SOD2(Ac-SOD2)expression.Thirty-two mice were randomly divided into control(Con)group 2,Honokiol(HKL)group 2,LPS treated(LPS)group 2,and LPS+HKL treatment(HKL+LPS)group 2.Hematoxylin and eosin(HE)staining was used to observe pathological changes to the lung tissue.Results HKL pretreatment significantly reversed the LPS-induced increase in ROS and MDA levels(P＜0.05),SOD2 acetylation and Sirt3 down-regulation(P＜0.05).TUNEL and caspase analysis showed that HKL protected against the apoptosis of PMVECs induced by LPS.VE-cadherin fluorescence staining demonstrated that HKL pretreatment prevented LPS from disrupting cell adhesion junctions.Claudin-5 fluorescence staining showed that HKL pretreatment prevented LPS from disrupting the tight junctions between cells.In the animal experiments,HE staining showed that HKL significantly inhibited the typical pathological changes of ARDS in the lung tissue of mice in the LPS group.Conclusions HKL can significantly inhibit the LPS-induced oxidative stress,apoptosis,and cell-connection breakdown of PMVECs,thereby alleviating ARDS symptoms.

Objective To exploring the correlation between serum miR-195 and metabolic associated fatty liver disease(MAFLD)in type 2 diabetes mellitus(T2DM)patients.Methods A total of 79 patients with T2DM who were treated in the Endocrinology Department of Gansu Provincial People's Hospital were enrolled in this study from October 2022 to August 2023.They were divided into simple T2DM(n=37)and MAFLD group(MAFLD,n=42)according to whether they were complicated with MAFLD.Meanwhile,34 healthy individuals who underwent physical examinations were selected as the normal control(NC)group.Results Compared with the NC group,FPG,HbA1c and HOMA-IR increased,while HDL-C decreased in T2DM and MAFLD group(P<0.05).The TG and hs-CRP levels were higher in MAFLD group than in NC and T2DM group(P<0.05).The serum miR-195 expression decreased sequentially(P<0.05),while FAS increased sequentially in NC,T2DM,and MAFLD group(P<0.05).Spearman correlation analysis showed that miR-195 was negatively correlated with FPG,HOMA-IR,hs-CRP,and FAS(P<0.05).Multiple linear regression analysis showed that HOMA-IR and FAS were the influencing factors for serum miR-195.Conclusions Down-regulation of serum miR-195 expression in patients with T2DM combined with MAFLD may be by glucose and lipid metabolism,IR and inflammatory response.

Objective Some colorectal cancer patients still face high recurrence rates and poor prognoses even after they have undergone the surgical treatment of radical resection.Identifying potential biochemical markers and therapeutic targets for the prognostic evaluation of patients undergoing radical resection of colorectal cancer is crucial for improving their clinical outcomes.Recently,it has been reported that the T cell immunoglobulin and mucin domain protein 3(Tim-3)and its ligand galactose lectin 9(galectin-9)play crucial roles in immune dysfunction caused by various tumors,such as colorectal cancer.However,their expressions,biological functions,and prognostic value in colorectal cancer are still unclear.This study aims to investigate the relationship between Tim-3 and galectin-9 expression levels and the clinicopathological characteristics and prognosis of patients undergoing radical resection of colorectal cancer.Methods A total of 171 patients who underwent radical resection of colorectal cancer at Chengdu Fifth People's Hospital between February 2018 and March 2019 were selected.Immunohistochemistry was performed to assess the expression levels of Tim-3 and galectin-9 in the cancer tissue samples and the paracancerous tissue samples of the patients.The relationship between Tim-3 and galectin-9 expression levels and the baseline clinical parameters of the patients was analyzed accordingly.Kaplan-Meier analysis was performed to assess the association between Tim-3 and galectin-9 expression levels and the relapse-free survival(RFS)and the overall survival(OS)of colorectal cancer patients.Cox regression analysis was conducted to identify factors associated with adverse prognosis in the patients.Results The immunohistochemical results showed that the high expression levels of Tim-3 and galectin-9 were observed in 70.18%(120/171)and 32.16%(55/171),respectively,of the colorectal cancer tissues,whereas the low expression levels were 29.82%(51/171)and 67.84%(116/171),respectively.Furthermore,the expression score of Tim-3 was significantly higher in colorectal cancer tissues than that in the paracancerous tissues,while the expression score of galectin-9 was lower than that in the paracancerous tissues(P＜0.05).Further analysis revealed that the expression of Tim-3 and galectin-9 was associated with the depth of tumor infiltration,vascular infiltration,and clinical staging(P＜0.05).During the follow-up period of 14-63 months,7 out of 171 patients were lost to follow-up.Among the remaining patients,49 and 112 cases presented abnormally low expression of Tim-3 and galectin-9,respectively,whereas 115 and 52 cases presented high expression of Tim-3 and galectin-9,respectively.Kaplan-Meier survival analysis demonstrated that patients with high Tim-3 expression in colorectal cancer tissues had significantly lower RFS and OS than those with low expression did(RFS:log-rank=22.66,P＜0.001;OS:log-rank=19.71,P＜0.001).Conversely,patients with low galectin-9 expression had significantly lower RFS and OS than those with high expression did(RFS:log-rank=19.45,P＜0.001;OS:log-rank=22.24,P＜0.001).Cox multivariate analysis indicated that TNM stage Ⅲ(HR=2.26,95%CI:1.20-5.68),high expression of Tim-3(HR=0.80,95%CI:0.33-0.91),and low expression of galectin-9(HR=1.80,95%CI:1.33-4.70)were independent risk factors affecting RFS and OS in patients(P＜0.05).Conclusion Aberrant expression of Tim-3 and galectin-9 is observed in colorectal cancer tissues.High expression of Tim-3 and low expression of galectin-9 are closely associated with adverse clinico-pathological characteristics and prognosis.They are identified as independent influencing factors that may trigger adverse prognostic events in patients.These findings suggest that Tim-3 and galectin-9 have potential as new therapeutic targets and clinical indicators.