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.

Cancer stem cells (CSCs) are widely acknowledged as primary mediators to the initiation and progression of tumors. The association between microbial infection and cancer stemness has garnered considerable scholarly interest in recent years. Porphyromonas gingivalis (P. gingivalis) is increasingly considered to be closely related to the development of oral squamous cell carcinoma (OSCC). Nevertheless, the role of P. gingivalis in the stemness of OSCC cells remains uncertain. Herein, we showed that P. gingivalis was positively correlated with CSC markers expression in human OSCC specimens, promoted the stemness and tumorigenicity of OSCC cells, and enhanced tumor formation in nude mice. Mechanistically, P. gingivalis increased lipid synthesis in OSCC cells by upregulating the expression of stearoyl-CoA desaturase 1 (SCD1) expression, a key enzyme involved in lipid metabolism, which ultimately resulted in enhanced acquisition of stemness. Moreover, SCD1 suppression attenuated P. gingivalis-induced stemness of OSCC cells, including CSCs markers expression, sphere formation ability, chemoresistance, and tumor growth, in OSCC cells both in vitro and in vivo. Additionally, upregulation of SCD1 in P. gingivalis-infected OSCC cells was associated with the expression of KLF5, and that was modulated by P. gingivalis-activated NOD1 signaling. Taken together, these findings highlight the importance of SCD1-dependent lipid synthesis in P. gingivalis-induced stemness acquisition in OSCC cells, suggest that the NOD1/KLF5 axis may play a key role in regulating SCD1 expression and provide a molecular basis for targeting SCD1 as a new option for attenuating OSCC cells stemness.
Porphyromonas gingivalis/pathogenicity* ; Stearoyl-CoA Desaturase/metabolism* ; Humans ; Carcinoma, Squamous Cell/pathology* ; Mouth Neoplasms/metabolism* ; Animals ; Neoplastic Stem Cells/microbiology* ; Mice, Nude ; Mice ; Nod1 Signaling Adaptor Protein/metabolism* ; Kruppel-Like Transcription Factors/metabolism* ; Cell Line, Tumor

Innate lymphoid cells are an important part of innate mucosal immunity and participate in immune response by secreting effector cytokines and regulating the functions of other immune cells.They are similar to T helper cells in transcription factors and secretory cytokines,but they also have some unique functions.Different innate lymphocyte subsets play different roles in chronic obstructive pulmonary disease(COPD)according to their respective characteristics.Compared with adaptive lymphocyte,innate lymphocyte is relatively less in lymphoid tissue,accounting for only a small part of lung immune cells,but these lymphocyte cells play a crucial role in the development and progression of COPD.In this paper,the role of innate lymphoid cells in COPD and the related research progress were reviewed.

Objective To investigate the correlation of plasma homocysteine(Hcy)level and the polymorphisms of its key metabolic enzymes methylenetetrahy-drofolate reductase(MTHFR)gene with ischemic stroke(IS).Methods A total of 310 patients with IS were enrolled as the case group and 330 healthy subjects during the same period were selected as the control group.Plasma Hcy concentration was detected by enzyme cycling method,and the real-time fluorescent quantitative PCR(RT-qPCR)was used to detecte the genotypes of MTHFR C677T.Results The frequencies of TT genotype(36.13%),CT genotype(10.00%)and T allele(28.06%)of MTHFR gene C677T locus in stroke patients were significantly higher than that in control group(P＜0.05).The frequency of the TT genotype was significantly higher in IS group compared to control group,indicating a recessive mode of inheritance(P＜0.05);In the dominant mode of inheritance,the frequency of CT+TT genotype in IS group was also significantly higher than that in control group(P＜0.05);The plasma Hay concentration of MTHFR C677T genotype TT,CT and CC patients was statistically different(P＜0.05),which were(20.91±6.78)μmol/L(17.20±5.39)μmol/L,(14.35±4.32)μmol/L,respectively;The area under the ROC curve(AUC)of plas-ma total Hcy level was 0.610(95%CI:0.566～0.653,P＜0.001).It indicated that it might play an impor-tant role in predicting the risk of suffering from IS.Multivariate Logistic regression analysis showed that plasma Hcy level and MTHFR C677T gene polymorphism were important risk factors of IS.Conclusions Elevated plasma Hcy level is associated with IS,and the synergistic effect of elevated Hcy level and MTHFR C677T gene mutation may increase the risk of IS.

Objective To investigate the relationship between methylenetetra-hydrofolatereductase(MTHFR)gene polymorphism and serum homocysteine(Hcy)level and subtypes of ischemic stroke(IS).Methods The study was conducted according to the matched principle of case-control design,310 patients with IS and 330 healthy people during the same period were selected as the case group and the control group.Recycling enzyme method and the real-time fluorescent quantitative PCR(RT-qPCR)method were used to detect the level of serum Hcy and the genotypes of MTHFR C677T,respectively.Results There was a significantly difference in MTHFR C677T genotype and allele frequency between the case and control groups(P＜0.05).The correlation analysis with different subtypes indicated that the frequencies of CT genotype(38.02%),TT genotype(10.74%),and T allele(29.75%)were significantly different in the LAA group(OR=1.662,95%CI:1.058-2.608,P＜0.05;OR=2.373,95%CI:1.110-5.073,P＜0.05;OR=1.663,95%CI:1.190-2.323,P＜0.05);The frequencies of TT genotype(10.53%)and T allele(27.30%)in SAO group were also significantly different(OR=2.130,95%CI:1.046-4.336,P＜0.05;OR=1.474,95%CI:1.075-2.021,P＜0.05).Further analysis of serum Hcy level showed that LAA group(19.55±5.61)μmol/L and SAO group(16.37±5.20)μmol/L were significantly higher than the control group(14.46±4.61)μmol/L(P＜0.001);Among the patients of both subtypes the serum Hcy levels in those with CT genotypes and TT genotypes were significantly higher than those in patients of CC genotypes(P＜0.001).Conclusions The gene polymorphism of MTHFR C677T has a significant effect on Hcy level in pa-tients with LAA and SAO stroke.

Objective To investigate the value of cranial CT cisternal grading combined with D-dimer(D-D)and Glasgow Coma Scale(GCS)score in predicting the short-term postoperative prog-nosis of patients with severe traumatic brain injury.Methods A total of 165 patients with severe trau-matic brain injury who were treated in the hospital from January 2019 to May 2024 were selected as study subjects,all underwent craniotomy surgery.Postoperative follow-up was conducted for 3 months to analyze the differences in clinical data and preoperative indicators such as cranial CT cisternal grad-ing,D-D levels,and GCS scores between patients with poor and good prognosis.The value of cranial CT cisternal grading,D-D levels,and GCS scores in predicting short-term postoperative poor prognosis in patients with severe traumatic brain injury was also analyzed.Results Compared with patients with good prognosis,patients with poor prognosis had higher proportion of age,cranial CT cisternal grading of Ⅰ to Ⅱ,D-D levels,and GCS scores＜6(P＜0.05).There were no statistically significant differences in C-reactive protein,prothrombin time,activated partial thromboplastin time,international normalized ratio,total cholesterol,triglycerides,high-density lipoprotein cholesterol,and low-density lipoprotein cholesterol levels between patients with poor and good prognosis(P＞0.05).Cranial CT cisternal grading,D-D levels,and GCS scores were influencing factors for short-term postoperative poor prognosis in patients with severe traumatic brain injury(P＜0.05).The area under the curve for poor prognosis by three indicators in combination was 0.941(95％CI,0.906 to 0.975),which was higher than the area under the curve for the individual predictions of cranial CT cisternal grad-ing,D-D levels,and GCS scores(P＜0.05).Conclusion The influencing factors for short-term postoperative prognosis in patients with severe traumatic brain injury include cranial CT cisternal grading,D-D levels,and GCS scores.The model based on these three indicators has certain appli-cation value in predicting patient prognosis.

Objective To explore the effects of polystyrene microplastics(PS-MPs)on the growth,activity,oxida-tive stress levels,biofilm formation and virulence of Klebsiella pneumoniae.Methods Klebsiella pneumoniae was exposed to PS-MPs at different concentrations(10,50 and 100 μg/ml)and particle sizes(0.1,1.0 and 5.0 μm),and the growth curves were measured.The bacterial activity was determined by CCK-8(cell counting kit-8).The level of intracellular reactive oxygen species(ROS)was determined by fluorescence probe.The biofilm forming ability was determined by crystal violet staining.Real-time quantitative fluorescent PCR(qRT-PCR)was used to detect the relative expression levels of biofilm-forming genes(luxS,mrkA,wbbM,pgaA,wzm)and viru-lence genes(ureA,uge,wabG,fimH).Results A high concentration(100 μg/ml)of 0.1 μm PS-MPs had a stronger inhibitory effect on the growth and activity of Klebsiella pneumoniae,and the intracellular ROS level signifi-cantly increased,indicating that smaller particle size and higher concentration of PS-MPs were more toxic to bacte-ria.PS-MPs of 100 μg/ml particle size groups(0.1,1.0 and 5.0 μm)significantly promoted the biofilm forma-tion of Klebsiella pneumoniae.The relative expression levels of biofilm formation related genes(luxS,mrkA,wbbM,pgaA,wzm)and virulence genes(ureA,uge,wabG,fimH)increased.Conclusion By inducing Kleb-siella pneumoniae to produce a high level of ROS,PS-MPs can cause oxidative stress,inhibit the growth and activi-ty of bacteria,and enhance the biofilm formation ability and virulence,thus affecting the biological characteristics of Klebsiae pneumoniae.

Peripheral pulmonary lesions(PPL),including peripheral pulmonary nodules,are common lung problems.As the increase of patients with lung nodules,the demand for tissue sampling also increases.Safe and accurate biopsy techniques are very important for patients to identify benign and malignant lesions.Electronic bronchoscopy is one of the biopsy techniques for the diagnosis of PPL in recent decades.Various guiding techniques,such as radial probe endobronchial ultrasound and virtual navigation bronchoscope,have been proved to improve the performance of conventional bronchoscopy.This paper aims to provide an review of the available data on advanced bronchoscopic techniques and explore their application in diagnosing PPL.

Gallbladder cancer constitutes the most prevalent malignant tumor of the biliary tract. It exhibits clinical characteristics such as challenging diagnosis in early stages, aggressive invasiveness, and poor prognosis. Studies have discovered that the occurrence, development, and metastasis of malignant tumors are closely associated with the inflammatory response, and the level of systemic inflammation within the body can be reflected by peripheral blood inflammatory indicators. This review delineates the research progress on the correlation between peripheral blood inflammatory markers and the prognosis of gallbladder cancer, with the intention of providing references for the diagnosis, treatment, and prognosis evaluation of gallbladder cancer.