Objective To investigate the regulatory role of the programmed cell death protein 1 (PD-1) and its ligand programmed cell death protein ligand 1 (PD-L1) signaling on the subtypes and functions of natural killer (NK) cells at the maternal-fetal interface during the second trimester in mice following Toxoplasma gondii infection during the first trimester. Methods Twelve 6- to 8-week-old female mice of the C57BL/6J strain were divided into a control group and an infection group, of 6 mice in each group. On the 6.5th day of pregnancy (Gd6.5), each pregnant mouse in the infection group was intraperitoneally injected with 150 tachyzoites of the Toxoplasma gondii PRU strain, while mice in the control group were injected with an equal volume of physiological saline. On the 12.5th day of pregnancy (Gd12.5), uterus and placenta tissues were sampled from pregnant mice for pathological observations, and the mRNA expression levels of PD-1, PD-L1, and tumor necrosis factor-α (TNF-α) were quantified in uterus and placenta tissues. The PD-1 and DX5 expression was measured on NK cells at the maternal-fetal interface using flow cytometry. In addition, the in vitro JEG-3 trophoblast cells and NK-92MI cells co-culture system was established as the control group, and the addition of T. gondii tachyzoites in the co-culture system served as the infection group. The PD-1, PD-L1, and DX5 mRNA expression was quantified in cells using real-time fluorescence quantitative reverse transcription PCR (RT-qPCR) assay, and the TNF-α concentration was measured in the cell culture supernatant using enzyme-linked immunosorbent assay (ELISA). Results On Gd12.5, clear and intact cellular structures of placental decidual tissues were seen in pregnant mice in the control group, with no remarkable abnormal changes found in the uterine columnar epithelial cells, and inflammatory cell infiltration and blood stasis at varying degrees were found in uterine and placental tissues from pregnant mice in the infection group. The relative PD-1, PD-L1, and TNF-α mRNA expression was (1.004 ± 0.004), (1.001 ± 0.001), and (1.001 ± 0.001) in uterine tissues from pregnant mice in the control group and (2.480 ± 0.720), (3.355 ± 0.920), and (2.391 ± 0.073) in the infection group, respectively. The relative PD-1, PD-L1, and TNF-α mRNA expression was (1.007 ± 0.010), (1.006 ± 0.006), and (1.001 ± 0.001) in the uterine tissues in the control group and (6.948 ± 1.918), (3.225 ± 1.034), and (1.536 ± 0.150) in the infection group, respectively. The relative PD-1, PD-L1, and TNF-α mRNA expression was higher in both the uterine (t = 3.55, 4.43 and 33.02, all P values < 0.05) and placental tissues (t = 5.36, 3.72 and 6.18, all P values < 0.05) in the infection group than in the control group. Flow cytometry showed that the proportions of PD-1⁺ NK cells, PD-1⁺ DX5⁺ NK cells, and DX5⁺ NK cells were (12.200 ± 1.082)%, (9.373 ± 7.728)%, and (44.000 ± 4.095)% in uterine tissues from pregnant mice in the control group, and (21.733 ± 1.630)%, (18.767 ± 1.242)%, and (73.367 ± 0.611)% in the infection group, respectively. The proportions of PD-1⁺ NK cells, PD-1⁺ DX5⁺ NK cells, and DX5⁺ NK cells were (1.100 ± 0.510)%, (2.277 ± 1.337)%, and (96.167 ± 2.831)% in placental tissues from mice in the control group, and (26.867 ± 9.722)%, (23.433 ± 6.983)%, and (82.467 ± 2.248)% in the infection group, respectively. The proportions of PD-1⁺ NK cells (t = 8.45, P < 0.05) and DX5⁺ NK cells (t = 12.29, P < 0.05) were higher in uterine tissues from pregnant mice in the infection group than in the control group, and no significant difference was seen in the proportion of PD-1⁺ DX5⁺ NK cells (Z = -1.09, P > 0.05). The proportions of PD-1⁺ NK cells (t = 4.58, P < 0.05) and PD-1⁺ DX5⁺ NK cells (t = 5.15, P < 0.05) were higher in placental tissues from pregnant mice in the infection group than in the control group, while the proportion of DX5⁺ NK cells was lower in the infection group than in the control group (t = -6.56, P < 0.05). RT-qPCR assay revealed that the relative PD-1, PD-L1, and DX5 mRNA expression was (1.010 ± 0.005), (1.002 ± 0.003), and (1.001 ± 0.001) in the JEG-3 cells and NK92MI cells co-culture system and (3.638 ± 1.258), (0.397 ± 0.158), and (4.267 ± 1.750) in the control group, and ELISA measured that the TNF-α concentration was higher in the cell culture supernatant in the infection group [(22.056 ± 3.205) pg/mL] than in the control group [(12.441 ± 0.001) pg/mL] (t = 5.20, P < 0.05). The PD-1(t = 3.62, P < 0.05) and DX5 mRNA expression (t = 3.23, P < 0.05) was higher in the infection group than in the control group, and the PD-L1 mRNA expression was lower in the infection group than in the control group (t = -6.63, P < 0.05). Conclusions Following T. gondii infection, both PD-L1 expression and PD-1 expression on DX5⁺ NK cells at the maternal-fetal interface are upregulated in mice during the second trimester; however, the proportion of DX5⁺ NK cells decreases. These findings suggest that PD-1/PD-L1 signaling may suppress NK cell functions by modulating DX5⁺ NK cell subsets.

OBJECTIVES: To investigate the role of activating transcription factor 3 (ATF3) in atherosclerotic plaques for regulating inflammatory responses during atherosclerosis (AS) progression. METHODS: Human coronary artery specimens from autopsy cases were examined for ATF3 protein expression and localization using immunofluorescence staining and Western blotting. Apolipoprotein E-deficient (ApoE^-/-) mouse models of AS induced by high-fat diet (HFD) feeding for 12 weeks were subjected to tail vein injection of adeno-associated virus serotype 9 (AAV9) to knock down ATF3 expression. After an additional 5 weeks of HFD feeding, the mice were euthanized for analyzing structural changes of the aortic plaques, and the expression levels of ATF3, inflammatory factors (CD45, CD68, IL-1β, and TNF-α), and NF-κB pathway proteins (P-IKKα/β and P-NF-κB p65) were detected. In the cell experiment, THP-1-derived foam cells were transfected with an ATF3-overexpressing plasmid or an ATF3-specific siRNA to validate the relationship between ATF3 and NF‑κB signaling. RESULTS: In human atherosclerotic plaques, ATF3 expression was significantly elevated and partially co-localized with CD68. ATF3 knockout in ApoE^-/- mice significantly increased aortic plaque volume, upregulated the inflammatory factors, enhanced phosphorylation of the NF‑κB pathway proteins, and increased the expressions of VCAM1, MMP9, and MMP2 in the plaques. In THP-1-derived foam cells, ATF3 silencing caused activation of the NF‑κB pathway, while ATF3 overexpression suppressed the activity of the NF-κB pathway. CONCLUSIONS AS promotes ATF3 expression, and ATF3 deficiency exacerbates AS progression by enhancing plaque inflammation via activating the NF-κB pathway, suggesting the potential of ATF3 as a therapeutic target for AS.
Animals ; Activating Transcription Factor 3/metabolism* ; Signal Transduction ; NF-kappa B/metabolism* ; Humans ; Mice ; Plaque, Atherosclerotic/metabolism* ; Inflammation/metabolism* ; Apolipoproteins E ; Atherosclerosis/metabolism* ; Diet, High-Fat

Objective:To compare the clinical efficacy of laparoscopic Miles surgery through extraperitoneal stoma and intraperitoneal stoma.Methods:The medical records of 140 patients with low rectal cancer after laparoscopic Miles surgery admitted to Gastrointestinal Surgery of Northern Jiangsu People′s Hospital of Jiangsu Province from January 2018 to December 2022 were retrospectively analyzed. Among them, 80 were males and 60 were females, aged 50 to 75 years old, with an average age of 63.95 years old. They were divided into observation group (extraperitoneal stoma, n=70) and control group (intraperitoneal stoma, n=70) based on the stoma method. Through telephone, WeChat, outpatient follow-up and other contact methods, the intraoperative and postoperative recovery, the incidence of perioperative complications (stoma edema, stoma ischemia, peristoma inflammation, perineal/pelvic infection, lung infection) and the incidence of complications at 6 months and 1 year after surgery (stoma stricture, parastoma hernia/internal hernia, stoma prolapse/retraction), and the difference in the ability of artificial anus to control defecation at 1 year after surgery were compared between the two groups. SPSS27.0 statistical software was used for data analysis and processing. Results:(1) Incidence of individual complications such as lung infection between the two groups of patients during the perioperative period (4.3% vs 4.3%, χ2=0.17, P=0.676), stoma edema (25.7% vs 21.4%, χ2=0.36, P=0.550), stoma ischemia (7.1% vs 7.1%, χ2=0.00, P=1.000), peristomal inflammation (20.0% vs. 18.6%, χ2=0.05, P=0.830), perineal/pelvic infection (15.7% vs 27.1%, χ2=2.72, P=0.099), there was no difference between the two groups. There was still no difference in the overall complication rate between the two groups (72.9% vs 78.6%, χ2=0.62, P=0.430). (2) After follow-up to 6 months after surgery, the overall complication rate was 5.7% in the observation group compared with 22.9% in the control group ( χ2=7.06, P=0.008). In particular, the incidence of post-operative parastomal hernia/internal hernia did not occur in the observation group, while 8.6% of patients in the control group occurred (18.6% vs 42.9%, χ2=4.35, P=0.037). (3) After follow-up to 1 year after surgery, the overall complication rate in the observation group was lower than that in the control group ( χ2=8.59, P=0.003). The incidence of parastomal hernia/internal hernia after operation in the observation group was lower than that in the control group (2.9% vs 14.3%, χ2=4.47, P=0.034). (4) At the one-year follow-up, the overall excellent and good rate in the evaluation of bowel function in the observation group was higher than that in the control group (71.4% vs 48.6%, χ2=7.62, P=0.006). Conclusions:In laparoscopic Miles surgery for patients with rectal cancer, choosing extraperitoneal stoma has achieved good results, which can reduce the risk of complications 6 months or even 1 year after surgery, especially in preventing and controlling parastomal hernia/internal hernia. It has significant advantages, and at the same time, it can also promote the recovery of patients′ bowel function and reduce other related complications, thereby ensuring patient safety.

Objective To investigate the regulatory role and mechanism of mitochondrial riboso-mal protein S35(MRPS35)in the proliferation,invasion,and migration of colon cancer cells.Meth-ods A total of 120 colon cancer tissues and adjacent normal tissues from patients undergoing radical resection for colon cancer were collected.Human colon cancer cell lines(HCT116,SW480,SW620)and a human normal colon epithelial cell line(NCM460)were cultured.Bioinformatics analysis,real-time quantitative polymerase chain reaction,Western blot,immunohistochemical(IHC)analysis,and cellular functional experiments(plate clone formation assay,scratch test,Transwell migration assay,CCK-8 cell viability assay)were conducted to evaluate the expression and regulatory mechanism of MRPS35 in colon cancer.Results Bioinformatics analysis showed that the expression level of the MRPS35 gene was higher in colorectal cancer tissues than in adjacent normal tissues(P＜0.05).The relative expression levels of MRPS35 mRNA and MRPS35 protein were higher in human colon cancer cell lines(HCT116,SW480,SW620)than in NCM460 cells(P＜0.05).The relative ex-pression level of MRPS35 protein was higher in colon cancer tissues than that in adjacent normal tis-sues(P＜0.05).The expression level of MRPS35 was significantly correlated with tumor diameter,tumor differentiation,and T stage(P=0.002,0.021,0.036).Patients with high MRPS35 expres-sion had a higher overall survival rate than those with low MRPS35 expression(Log-rank P=0.015).After knockdown of MRPS35,the abilities of colon cancer cell cloning,proliferation,invasion,and migration were significantly enhanced.Furthermore,the expression of Wnt1,β-Catenin,and their downstream target proteins increased significantly after MRPS35 knockdown.Conclusion MRPS35 is significantly overexpressed in both colon cancer tissues and colon cancer cells,and it may inhibit the occurrence and development of colon cancer by regulating the Wnt/β-Catenin signaling pathway.Therefore,MRPS35 has the potential to become a novel biomarker and therapeutic target for colon cancer.

AIM:To investigate the impact of total flavonoids of Pterocarya hupehensis Skan(PHSTF)on the migration,invasion,and ferroptosis of non-small-cell lung cancer A549 cells.METHODS:The A549 cells were divided into control group,low-,medium-and high-dose(100,150 and 200 μg/mL)PHSTF groups,ferroptosis inhibitor liprox-statin-1(Lip-1)group,and high-dose PHSTF combined with Lip-1 group,each cultured in corresponding media.Cell via-bility was assessed using the CCK-8 assay,while cell migration and invasion abilities were determined through scratch and Transwell assays.Cell lipid peroxidation levels were measured using the glutathione(GSH)assay kit.RT-qPCR was em-ployed to assess the mRNA expression of solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4),while Western blot was utilized to examine the protein expression of SLC7A11,GPX4,Kelch-like epichlorohy-drin-associated protein-1(Keap-1),nuclear factor E2-related factor 2(Nrf2)and heme oxygenase-1(HO-1).RE-SULTS:Compared with control group,PHSTF significantly diminished the viability of A549 cells in a time-and dose-de-pendent manner(P＜0.01),and the cell migration and invasion were also reduced(P＜0.01),along with a significant de-crease in GSH level(P＜0.01).Treatment with PHSTF inhibited the mRNA and protein expression levels of ferroptosis-re-lated proteins,including SLC7A11 and GPX4(P＜0.01),suppressed the protein expression of Nrf2 and HO-1(P＜0.01),and enhanced the expression of Keap-1(P＜0.01).The Lip-1 partially restored the decrease in cell viability in-duced by PHSTF(P＜0.01),significantly up-regulated the protein expression levels of SLC7A11,GPX4,Nrf2 and HO-1,and suppressed the protein expression of Keap-1(P＜0.01).CONCLUSION:Total flavonoids of Pterocarya hupehen-sis Skan can inhibit the migration and invasion of non-small-cell lung cancer A549 cells,and induce the cell ferroptosis by regulating the Keap-1/Nrf2/HO-1 pathway.

Objective To explore the effect of histologic chorioamnionitis(HCA)on clinical outcomes of preterm infants with a gestational age<34 weeks.Methods This retrospective study enrolled 497 cases of premature infants with a gestational age<34 weeks and their mothers who were hospitalized in the Qingdao Women and Children's Hospital from January 2019 to December 2022.According to whether the pathology of placenta was diagnosed as HCA or not,patients were divided into the HCA group(257 cases)and the control group(240 cases).The propensity score matching analysis was performed at a ratio of 1︰1.Ten items were matched,including gestational age,birth weight,gender,cesarean section,gestational diabetes mellitus,gestational hypertension,placental abruption,premature rupture of membranes,use of antenatal glucocorticoids and assisted reproductive technology.The differences of major complications and survival rate were compared between the two groups.Results A total of 156 pairs premature infants were successfully matched.Before matching,the incidences of early-onset sepsis(EOS)and bronchopulmonary dysplasia(BPD)were higher in the HCA group than those of the control group(26.1%vs.7.5%,45.1%vs.25.8%,P＜0.01).The incidence of EOS was higher in the HCA group than that of the control group after matching(24.4%vs.7.7%,P＜0.01),and the incidence of neonatal respiratory distress syndrome(NRDS)was significantly lower in the HCA group than that in the control group after matching(34.0%vs.46.8%,P＜0.05).There were no significant differences in survival rate and the incidences of other complications between the two groups before and after matching(P>0.05).Conclusion Preterm infants exposed to HCA have a higher risk of EOS and a lower risk of NRDS after propensity score matching.HCA has no significant effect on survival rate and other complications of premature infants.

Objective:To explore the clinical application and triage management value of using blood circulating cell-free DNA (cfDNA) (cysteine dioxygenase type 1 gene, CDO1, and Homeobox protein A9 gene, HOXA9) hypermethylation level to detect and diagnose ovarian cancer.Methods:A case-control study was conducted on patients who went for surgery at Chengdu Womens and Childrens Central Hospital from November 2022 to October 2023. Blood samples were collected before surgery for evaluation of cancer antigen 125 (CA125), human epididymis protein 4 (HE4), risk of ovarian malignancy algorithm (ROMA) score, and DNA methylation testing. The basic clinical information, biomarkers, and transvaginal ultrasound (TVS) information were collected simultaneously. Information from a total of 151 patients was collected, including 122 cases with benign pathology and 29 ovarian cancer cases. The pathologic diagnosis of ovarian tissue was defined as the gold standard. The multivariate logistic regression analysis was used to identify high-risk factors for ovarian cancer. The clinical efficacy of DNA methylation detection for ovarian cancer was analyzed using the area under curve (AUC).Results:The results showed that the age, menopausal status, CA125 and HE4 detection, ROMA score, positivity rate of CDO1 gene and HOXA9 gene single or combined testing in ovarian cancer patients were higher than those in the benign group and showed significant differences ( P<0.05). Among these detection protocols, the AUC of CDO1 and HOXA9 dual gene methylation testing for ovarian cancer was the highest at 0.936 (95% CI, 0.878-0.994), with 89.7% (95% CI 73.6%-96.4%) sensitivity and 97.5% (95% CI 93.0%-99.2%) specificity, respectively. The positive detection rate of CDO1 and HOXA9 dual gene methylation in early ovarian cancer FOGO I-II stage is 12/14 higher than other tests. Conclusion:Blood cfDNA methylation detection, a simple, non-invasive, and highly sensitive detection method, is superior to the current ovarian cancer testing in the risk assessment and early detection.

  Objective  To promote the development of high-quality health standards in China and summarize the characteristics of global health standards management systems and development processes.  Methods  We conducted a systematic search of relevant databases and the websites of standardization institutions. A descriptive method was used to analyze the basic characteristics of the standard management systems and development processes.  Results  The majority of the investigated countries and organizations primarily implement voluntary standards, with the exception of Chinese and the European Committee for Standardization that enforce compulsory standards. All countries and organizations have health standards at the national, group, and enterprise levels. In China and Japan, standards authorities are subordinate to the government, while in other countries or organizations, they operate as independent civil societies. The development process for health standards generally involves five stages: "Project/Proposal → Draft → Enquiry → Review → Publication". However, the specific steps in the development of health standards vary across different countries and organizations. Furthermore, China ranks high in the number of both standards and health standards issued, but the proportion of health standards is only 2.06%.  Conclusions  There are significant differences between China and other countries in terms of the management system and development process of health standards. To promote the management and development of high-quality health standards in China, we recommend promoting the coordination mechanism for standard development, increasing efforts to publicize standards and evaluate the implementation effect, strengthening the construction of personnel for the development of health standards, optimizing the efficiency of standard development and improving the internationalization level of health standards.

National health standards involve all kinds of technical requirements formulated and numbered in accordance with the procedures and formats stipulated in the standardisation system for the implementation of national health and hygiene laws, regulations and policies, and the protection of human health. The establishment of health standards in China should align with our legal framework, including laws, regulations, departmental rules, and health and hygiene policies. During the development of these standards, a comprehensive approach is advocated, encompassing in-depth investigations, rigorous demonstrations, and extensive stakeholder engagement. However, the process of standard formulation may suffer from insufficient research evidence. The evidence-based concept emphasizes the significance of evidence. Therefore, integrating evidence-based concept into the process of developing health standards can enhance the quality and scientific basis of these standards. This article systematically elucidates the current status and development process of health standards in China, explores the necessity and feasibility of incorporating evidence-based concept into the development of these standards, analyzes the challenges and opportunities, and presents reflections and suggestions.

Objective:To observe the clinical features, treatment and prognosis of ciliary body tumors.Methods:A retrospective clinical study. From November 2011 to March 2023, 8 cases (8 eyes) with ciliary body tumours confirmed by pathohistological examination at the Department of Ocular Oncology, Beijing Tongren Hospital were included in the study. Patients' age, gender, involved eyes, symptoms, best corrected visual acuity (BCVA), intraocular pressure, cataract, lens subluxation, and imaging manifestations were collected in detail. All affected eyes were treated surgically. The follow-up time after surgery ranged from 1 to 10 years. The patients' clinical presentation as well as imaging, pathohistological features and treatment and prognosis were analysed retrospectively.Results:Among 8 cases (8 eyes), there were 3 males (3 eyes) and 5 females (5 eyes), 3 and 5 eyes in the right and left eyes, respectively. The median age was 44 years. Ciliary body medulloepitheliomas, melanoma, squamous cell carcinoma, leiomyoma, schwannoma, and adenoma of the nonpigmcnted ciliary epithelium were in 2, 2, 1, 1, 1, and 1 eyes, respectively. All reported decreased or loss of vision. Cataract, vitreous opacity, red eye and or (ocular pain), retinal detachment, lens subluxation, and secondary glaucoma were 6, 4, 4, 2, 1, and 1 eyes, respectively. Diagnostic imaging was consistent with pathological findings in 3 eyes. The first surgery was performed for enucleation and orbital implantation in 2 eyes, the patients were 9 and 10 years old with medullary epithelioma; the follow-up time after surgery was 1 and 5 years, respectively. Local tumour resection was performed in 6 eyes. Among them, 3 eyes with benign tumours were followed up for 1 to 9 years after surgery; 2 eyes showed significant improvement in visual acuity, 1 eye with adenoma of the nonpigmcnted ciliary epithelium had a preoperative BCVA of finger count/1 m, and a postoperative BCVA of 0.5, and 1 eye with leiomyoma had a preoperative BCVA of 0.06, and a postoperative BCVA of 0.5; and 1 eye was lost to follow-up. Malignant tumour in 3 eyes, of which 2 eyes recurred after surgery. Re-operation for enucleation and local tumour excision combined with local cryotherapy in 2 eyes of recurrence were 1 eye each, respectively. The follow-up period after surgery was 2 and 4 years, respectively. No recurrence after surgery in 1 eye, but there was no significant improvement in visual acuity during follow-up. No recurrence or metastasis was observed in any of the eyes during the follow-up period or at the final follow-up.Conclusions:Ciliary body tumour types and clinical presentations are complex and varied; imaging can detect tumours but is poor at determining the nature of the lesion. Benign tumours do well with local excision surgery; malignant tumours do well with enucleation.