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

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.

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.

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.

Objective:To investigate the role of hepatitis B virus X protein (HBx) in glomerular podocyte immune disorder and its regulatory mechanism.Methods:Fourteen 6-week-old male hepatitis B virus (HBV) transgenic (HBV-Tg) mice were selected, and age-matched wild type (WT) mice were as controls. They were fed to different weeks, and 24 h urinary protein, blood biochemistry, renal pathology and podocyte changes under electron microscope were detected. The expression of HBx and the infiltration of immune cells in kidney tissue of HBV-Tg mice were observed by immunohistochemistry. Human podocyte cell line was transfected with pcDNA3.1/myc-HBx plasmid, and the localization of HBx and Nephrin in podocytes was detected by immunofluorescence. The expression of major histocompatibility complex Ⅱ (MHC-Ⅱ) and co- stimulatory molecule CD40 on the cell surface was detected by flow cytometry. The contents of multiple cytokines in cell culture supernatants were determined by enzyme-linked immunosorbent assay. Transcriptome sequencing (RNA-seq) was used to screen the downstream related genes regulated by HBx, and real-time quantitative PCR was used to verify their expressions. After overexpression or silencing of Notch1 gene with overexpressed plasmids or short hairpin RNA (shRNA) in podocytes, the effects on the expression of immune molecules and cytokines secretion was observed. The Notch receptor inhibitor N-[N-(3, 5-difluorophenyl-l- alanyl)]-(s)-phenylglycine tert-butyl ester (DAPT) was used to block Notch1 signaling pathway in HBV-Tg mice, and then blood biochemistry, renal pathological changes and infiltration of immune cells in kidney tissue were observed. Results:Twenty-four-hour urine protein, serum creatinine and urea nitrogen levels were markedly increased (all P<0.05) and renal pathological injury was significantly aggravated in HBV-Tg mice than those in WT mice. Also, HBx was up-regulated and immune cells infiltrated in the glomerulus of HBV-Tg mice. After transfection with HBx in podocytes, the expression of MHC-Ⅱ and CD40 on the cellular surface was up-regulated (all P<0.05), the contents of monocyte chemotactic protein-1 (MCP-1), tumor necrosis factor -α (TNF-α) and interleukin (IL)-1β in the supernatants were increased (all P<0.05), and the secretion of IL-4 and interferon γ (IFN-γ) was unbalanced. RNA-seq screened downstream genes of HBx, such as Notch1, PLA2R, TLR4, etc; and further confirmed that HBx could promote the up-regulation of Notch1 mRNA and protein (all P<0.05). After over-expression of Notch1 gene, HBx-induced expression of MHC-Ⅱ and CD40 on the cellular surface was significantly up-regulated (all P<0.05), and the contents of MCP-1, TNF-α and IL-1β in the supernatants were obviously increased (all P<0.05), and the imbalance of IL-4/IFN-γ was further aggravated. After Notch1 gene silencing, the above results showed the opposite changes. In vivo, the results indicated that serum creatinine levels were obviously decreased (all P<0.05), renal pathological injury and immune cell infiltration were significantly alleviated in HBV-Tg+DAPT group than those in HBV-Tg+DMSO group. Conclusions:HBx protein can promote the up-regulation of Notch1 signaling pathway in podocytes. And Notch1 signaling pathway promotes the expression of immune molecules on the surface of podocytes and regulates the imbalance of cytokines, then causes glomerular injury and dysfunction of immune microenvironment.

To improve the current situation of multiple preoperative visits and evaluations for elderly patients and other patients with complex conditions, in December 2022, Peking Union Medical College Hospital established preoperative multidisciplinary evaluation clinic (shorted as joint clinic). The joint clinic established a multidisciplinary team, clarified service targets, and developed standardized clinic workflows to provide patients with a " one-stop" preoperative assessment(physical fitness assessment, nutritional assessment, and frailty assessment, etc.), nutritional optimization intervention, and prerehabilitation education and guidance services. This practice strengthened preoperative risk management, improved preoperative assessment efficiency, and ensured the safety of patients during the perioperative period. As of September 2023, the joint clinic had received a total of 128 patients, of which 86 underwent surgery after preoperative evaluation and prehabilitation optimization. The obesity rate, smoking rate, and number of frailty cases of these patients had decreased from 13.96%, 11.63%, and 18 at the time of visit to 9.30%, 4.65%, and 14 on the day before surgery, respectively. They had recovered well after surgery. This practice had improved the preoperative status of patients and created conditions for high-risk patients to undergo surgery smoothly, so as to provide references for other hospitals to carry out multidisciplinary collaborative preoperative evaluation works.

As a new type of environmental pollutants, microplastics are widely distributed in the global ecosystem, and ingestion of microplastics may produce a number of toxic effects. Based on currently available publications, this paper describes the main pathways of exposure to microplastics, and summarizes the toxic mechanisms of microplastics in mammals, including oxidative stress, inflammatory response, immune damage, imbalance of gut microbiota, energy metabolism disorder and DNA damage, so as to provide insights into elucidation of the toxic mechanism mechanisms and health risk assessment of microplastics.

Pregnancy ; Female ; Humans ; Pregnancy Outcome ; Cross-Sectional Studies ; Lupus Erythematosus, Systemic/epidemiology* ; Pregnancy Complications/epidemiology* ; Menstruation Disturbances