Objective To explore the mechanism of malignant behavior of cervical cancer(CC)cells based on AMP-activated protein kinase(AMPK)/rapamycin target protein(mTOR)signaling pathway mediated by nucleo-tide-binding oligomerization domain receptor 2(NOD2).Methods Bioinformatics analysis was performed to deter-mine the expression of NOD2 in CC tissue.Plasmids targeting NOD2(shNOD2)and shRNAs negative control(shNC),NOD2 overexpression(NOD2)and vectors(Vec)were transfected into CC cells.The effect of NOD2 on the growth of CC cells was determined by cell counting kit-8 assay,colony formation and Transwell cell invasion as-say.Transcriptome analysis was performed by high throughput RNA sequencing(RNA-Seq).Western blot was used to detect the expression of NOD2,AMPK/mTOR signaling pathway and autophagy protein in the cell line.24 female BALB/c nude mice were randomly divided into four groups,with 6 mice in each group:vector group(Vec group),NOD2 overexpression group(NOD2 group),shNC group and shNOD2 group.The distant metastasis mod-el was established in mice,and the fluorescence intensity of lung metastasis was monitored and the number of lung metastasis nodules was counted.Results On-line database analysis showed that the expression of NOD2 in CC tis-sues was significantly higher than that in normal tissues,and there were significant differences in the mRNA expres-sion of NOD2 in different stages of CC(P＜0.05).In addition,the high expression of NOD2 was associated with poor overall survival and disease-free survival(P＜0.05).NOD2 overexpression promoted the proliferation,colony formation,migration and invasion of CC cells,while NOD2 knock-down was the opposite.Consistent with the re-sults in vitro,the lung colonization and lung metastasis of CC cells in NOD2 group were significantly higher than those in Vec group(P＜0.05),while those in shNOD2 group were significantly lower than those in shNC group(P＜0.05).RNA-Seq results showed that the expression of NOD2 was significantly related to AMPK signal activa-tion,mTOR signal inhibition,autophagy regulation pathway activation and autophagy formation.Compared with shNC group,the expression of phosphorylated AMPK and LC3 protein decreased significantly in shNOD2 group(P＜0.05),and the expression levels of phosphorylated mTOR and p62 protein increased significantly(P＜0.05).Compared with Vec group,the expression levels of LC3 and AMPK protein in NOD2 group increased significantly(P＜0.05),and the expression levels of phosphorylated mTOR and p62 protein decreased significantly(P＜0.05).Compared with shNC group,the point accumulation of GFP-mRFP-LC3 in shNOD2 group decreased signifi-cantly(P＜0.05).Compared with Vec group,the point accumulation of GFP-mRFP-LC3 increased significantly(P＜0.05).Conclusion NOD2 may promote the proliferation,migration and invasion of CC through AMPK/mTOR signal,and its mechanism partly involves autophagy activation.

Objective:To compare the characteristics of background mucosa, lesion features, and the efficiency of endoscopic submucosal dissection(ESD)between elderly and non-elderly patients with early gastric cancer(EGC).Methods:This study retrospectively collected data on patients with EGC who underwent ESD treatment at Beijing Hospital from April 2020 to December 2022.The clinical characteristics, background mucosa, lesion features, ESD outcomes, and pathological results of the patients were analyzed to compare the differences between elderly and non-elderly patients.Results:A total of 100 patients with EGC were selected, comprising 57 patients in the elderly group and 43 patients in the non-elderly group, with a total of 111 lesions identified(64 lesions in the elderly group and 47 lesions in the non-elderly group).The proportion of patients with a history of chronic atrophic gastritis was significantly higher in the elderly group(89.5%、51/57)compared to the non-elderly group(74.4%、32/43), with a statistically significant difference( P=0.047).Additionally, the difference in the extent of atrophy between elderly patients with EGC and their non-elderly counterparts was statistically significant( P=0.022).Among these patients, the proportion of those classified as Kimura-Takemoto C0 to C1 in the elderly group(15.6%、10/64)was lower than that in the non-elderly group(40.4%、19/47).In contrast, the proportion of patients classified as C2 to C3 in the elderly group(65.6%、42/64)was higher than that in the non-elderly group(51.1%、24/47), and the proportion of those classified as O1 to O3 in elderly patients(12.5%、8/64)was also higher than in the non-elderly group(4.3%、2/47).Furthermore, the difference in the extent of intestinal metaplasia between elderly and non-elderly patients with early gastric cancer was statistically significant( P=0.007).The overall proportion of total intestinal metaplasia in elderly patients(85.9%、55/64)was significantly higher than that in non-elderly patients(61.7%、29/47).Notably, the proportion of patients exhibiting extensive intestinal metaplasia(intestinal metaplasia present in both the gastric antrum and gastric body)was greater in the elderly group(43.8%、28/64)compared to the non-elderly group(23.4%、11/47).The Kyoto gastric cancer risk endoscopic score for elderly patients with EGC was(2.43±1.28)points, significantly higher than that of the non-elderly group(1.72±1.41)points, with a statistically significant difference observed( t=2.778, P=0.006).No statistically significant differences were observed in the proportions of total resection rates, R0 resections, curative resections, or postoperative complications following ESD when comparing elderly patients with EGC to their non-elderly counterparts. Conclusions:The proportion of extensive atrophy and intestinal metaplasia was higher in the background mucosa of elderly patients with EGC, and correspondingly, the Kyoto endoscopic gastric cancer risk score was elevated.Therefore, endoscopic examinations for elderly patients with chronic atrophic gastritis should be conducted with greater care and comprehensiveness.

Objective To investigate the predictive value of preoperative blood inflammatory markers for the recurrence risk in patients with basal cell carcinoma(BCC).Methods A total of 225 patients with BCC were divided into the high-risk recurrence group(155 cases)and the low-risk recurrence group(70 cases).General information and preoperative hematological indicators were collected in the two groups of patients.The neutrophil-to-lymphocyte ratio(NLR),lymphocyte-to-monocyte ratio(LMR),systemic inflammation marker(SIM)and platelet-to-lymphocyte ratio(PLR)were calculated.Receiver operating characteristic(ROC)curves were used to determine the predictive value of hematological markers with statistically significant differences between the two groups for BCC recurrence and to establish optimal cutoff values.Univariate and multivariate Logistic regression analyses were conducted to identify factors influencing BCC recurrence.A multivariate Logistic regression model was established to predict the recurrence risk of BCC.Area under the curve(AUC)and the Hosmer-Lemeshow test were used to evaluate the prediction efficiency and goodness-of-fit of the model.Results ROC analysis identified that optimal cutoff values for LMR and SIM were 5.12 and 0.86,respectively.Univariate Logistic regression analysis showed that LMR,SIM,ulceration at the primary tumor site,UV exposure and tumor maximum diameter were factors influencing BCC recurrence.Multivariate Logistic regression revealed that SIM≥0.86,tumor maximum diameter≥2.0 cm and UV exposure were risk factors for BCC recurrence,while LMR≥5.12 had a protective effect.The Logistic prediction model for BCC recurrence risk was Logit(P)=-1.598-1.517×LMR+1.323×SIM+2.406×UV exposure+3.465×tumor maximum diameter,with good model fit(P=0.725)and an AUC of 0.869(95%CI:0.822-0.917)for predicting BCC recurrence risk.Conclusion Monitoring preoperative LMR and SIM levels can assist in assessing the risk of recurrence in BCC patients and provide important guidance for clinical decision-making.

Objective To investigate the expression of IL-18,IL-18BP,and IL-18R in the peripheral blood B lymphocytes and mono-cytes of patients with atopic dermatitis(AD).Methods Peripheral venous blood was collected from 28 patients with AD and 21 healthy controls,and stimulated with Artemisia sieversiana wild allergen extract,house dust mite allergen extract,or Platanus pollen allergen extract.The expression of IL-18+,IL-18BP+,and IL-18R+in B lymphocytes and monocytes was measured using flow cytometry.Results Compared with the healthy control group,the proportions of IL-18+,IL-18BP+,and IL-18R+cells in the B lymphocyte group of patients with AD at rest increased 2.01-,10.35-,and 20.85-fold,respectively.The proportions of IL-18+and IL-18BP+cells in monocytes increased 5.51-and 41.88-fold,respectively,whereas the proportion of IL-18R+cells did not differ significantly between the groups.Conclusion IL-18,IL-18BP,and IL-18R in B lymphocytes and monocytes may play an important role in AD.IL-18,IL-18BP,and IL-18R may be potential targets for the treatment of AD.

Objective:To explore long-term outcome and risk factors of recurrence in stage Ⅲ gastric cancer patients who underwent radical gastrectomy and adjuvant chemotherapy.Methods:The clinical and pathological data of patients with stage Ⅲ (AJCC V8) gastric adenocarcinoma were analyzed retrospectively. All patients received radical gastrectomy and adjuvant chemotherapy consisting of oxaliplatin, fluoropyrimidines with or without docetaxel in our center during 2006 and 2011.Results:A total of 324 patients were enrolled into the study. With a median follow-up time of 108 months, 175 (54%) patients developed tumor recurrence. One hundred and eighty-three (56.5%) patients died, including 169 (52.2%) dying of gastric cancer recurrence. The median disease-free survival (DFS) was 35 months, and the median overall survival (OS) was 64 months. The 5-year OS rates were 58.2%, 51.5% and 25.6% in patients with stage ⅢA, ⅢB and ⅢC diseases, respectively ( P<0.01). Multivariate analysis revealed that T4b cancers ( P=0.02), higher lymph meta node ratio (LNR) ( P<0.01) and perineural invasion ( P=0.01) were independent negative prognostic factors, while more than 12 weeks of adjuvant chemotherapy may improve survival. Higher LNR was correlated with locoregional ( P<0.01), distant lymph node metastases ( P<0.01), and peritoneal metastases ( P=0.038). Perineural invasion ( P=0.047) was prone to peritoneal metastases. More than 12 weeks of adjuvant chemotherapy could reduce the risk of haematogenous metastases ( P=0.023). Conclusions:Outcomes were significantly different in subgroups of patients with stage Ⅲ gastric cancers after radical gastrectomy. Higher LNR and perineural invasion could predict poor prognosis and different recurrence patterns.

Abstract Non suicide self injury is highly common in adolescents, which is seriously threatening their physical and mental health. It is an important predictor of future suicide, and has become a focus of global public health concern. At present, the research on adolescent non suicidal self injury is still in its infancy, and its formation process is complex. The pathogenesis is not completely clear, and the relevant treatment studies are relatively few. The paper expounds the pathogenesis and treatment of the nonsuicidal adolescent NSSI from the perspectives of genetics, neurobiology, neuroimage and social psychology, aiming to provide a theoretical basis for adolescent NSSI prevention and intervention.

ObjectiveTo investigate the effect and mechanism of Nongsuo Dangguiwan in improving ovarian oxidative stress in rats with ovarian dysfunction. MethodThirty-six adult female SD rats were randomly divided into normal group， model group， positive drug group （Femoston， 0.3 mg·kg^-1）， and high-， medium-， and low-dose groups of concentrated Nongsuo Dangguiwan （2.08， 4.16， 8.32 g·kg^-1）， with six rats in each group. Rats， except for those in the normal group， were injected with 80 mg·kg^-1 vinyl cyclohexene dioxide （VCD） per day for 14 consecutive days to induce ovarian dysfunction. From the 15th day， rats were treated with corresponding drugs by gavage， while those in the model group received 2 mL·kg^-1 saline， once daily for 28 consecutive days. The ovarian index， levels of related hormones including estradiol （E₂）， follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and anti-mullerian hormone （AMH） in serum， as well as superoxide dismutase （SOD） activity and glutathione （GSH） content in serum were measured by enzyme-linked immunosorbent assay （ELISA）. The malondialdehyde （MDA） content in serum was detected by the thiobarbituric acid （TAB） method. Ovarian morphology was observed by hematoxylin-eosin （HE） staining. The expression of nuclear factor E₂-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， SOD2， and SOD1 in ovarian tissues was detected by immunohistochemistry （IHC） and Western blot. ResultCompared with the normal group， the model group showed a significant reduction in growing follicles in the ovary， loose arrangement of granulosa cells in the follicle， decreased body weight， ovarian index， and serum AMH and E₂ levels， increased LH and FSH levels （P<0.01）， reduced levels of SOD and GSH in serum （P<0.01）， and increased MDA level （P<0.01）. Compared with the model group， the groups with drug intervention showed increased ovarian index （P<0.05， P<0.01）， increased serum E₂ level （P<0.05， P<0.01）， decreased FSH， AMH， and LH levels （P<0.05， P<0.01）， increased number of growing follicles in the ovary， potentiated SOD activity in serum， increased GSH content， decreased MDA content （P<0.05， P<0.01）， and up-regulated expression levels of Nrf2， HO-1， SOD2， and SOD1 proteins in ovarian tissues （P<0.05， P<0.01）. ConclusionNongsuo Dangguiwan can regulate serum hormone levels， increase the expression of Nrf2， HO-1， SOD2， and SOD1 in ovarian tissues， and improve ovarian antioxidant capacity to resist oxidative stress injury， thereby improving ovarian reserve function.

Objective:To investigate the relationship of blood lipid levels with bone mass and fracture risk in elderly patients with type 2 diabetes mellitus (T2DM).Methods:A total of 744 elderly patients with T2DM who were treated in Tangshan Second Hospital from November 2018 to May 2020 were divided into normal bone mass group, low bone mass group and osteoporosis group according to bone mass levels. The total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and triglyceride (TG) levels in the three groups were compared, and the relationship between lipid indexes and bone mass was analyzed. The risk of fracture was calculated in the low bone mass group, and the relationship between lipid index and fracture risk was analyzed by linear regression. The blood lipid index between subjects with fracture and without fracture in osteoporosis group was compared, and the relationship between blood lipid index and fracture was analyzed by logistic regression.Results:There were significant differences in gender and age among the three groups (χ 2=38.80, F=4.94, P<0.05). The normal bone mass group had the smallest proportion of women and the youngest average age, while the osteoporosis group had the largest proportion of women and the average age. maximum. The LDL-C level in normal bone mass group was higher than those in the low bone mass group and the osteoporosis group, and LDL-C level in the low bone mass group was higher than that in the osteoporosis group ( F=3.38, P<0.05). In the low bone mass group, the risk of systemic fracture was 3.50% (2.40%, 4.10%) and hip fracture was 0.99% (0.80%, 1.20%). Linear regression showed that LDL-C and TG were positively correlated with the risk of systemic fractures in the low bone mass group (LDL-C: B=0.98, P=0.006;TG: B=0.23, P=0.024);TG was positively correlated with the risk of hip fracture in the low bone mass group ( B=0.16, P=0.002). In the osteoporosis group, the levels of HDL-C and LDL-C were lower in the patients with fractures than those without fractures ( t=3.24, P=0.001; t=2.98, P=0.003). Logistic regression analysis showed that higher HDL-C and LDL-C levels were protection factors for fracture risk in the osteoporosis group ( β=-2.73, P=0.009, OR=0.06, 95 %CI=0.04-0.10; β=-0.15, P=0.033, OR=0.83, 95 %CI=0.74-0.99). Conclusion:The relationship of serum lipid index with bone mass and fracture risk in hospitalized elderly T2DM patients is complicated, it is suggested to set individual blood lipid control targets according to the bone mass of patients.

ObjectiveTo explore the mechanism of Jianpi Yishen Huazhuo prescription in the improvement of ovarian function in polycystic ovary syndrome （PCOS）. MethodSeventy female SD rats in SPF grade were randomly divided into 6 groups， 15 in the blank group and 15 in the model group， 10 in the metformin group （0.1 g·kg^-1·d^-1）， and 10 in the low （1.275 g·kg^-1·d^-1）， medium （2.55 g·kg^-1·d^-1）， and high-dose （5.10 g·kg^-1·d^-1） Jianpi Yishen Huazhuo prescription groups. The blank group was given normal saline （10 mL·kg^-1·d^-1） by gavage and ordinary feed， and the other groups were given letrozole （1 mg·kg^-1·d^-1） by gavage combined with high-fat feed for 21 days to induce the model of PCOS. After modeling， the blank group and model group were given equal volume normal saline by gavage， and each drug group was given the corresponding dose of the drug by gavage for 30 days. The changes in body mass and fasting blood glucose （FPG） of rats before and after modeling were compared. Hematoxylin eosin （HE） staining was used to observe the morphological change in the ovaries of rats in each group. The serum levels of follicle stimulating hormone （FSH）， luteinizing hormone （LH）， testosterone （T）， anti-Mullerian hormone （AMH）， and estradiol （E₂） were measured by enzyme-linked immunosorbent assay （ELISA）， and the LH/FSH ratio was calculated. Immunohistochemical staining （IHC） and Western blot were used to detect the protein expression levels of nucleoside binding oligomerization domain protein like receptor 3 （NALP3）， apoptosis-associated speck-like protein （ASC）， cysteine protease-1 （Caspase-1）， nuclear transcription factor-κB （NF-κB）， interleukin-1β （IL-1β）， interleukin-18 （IL-18）， and interleukin-6 （IL-6） in the rat ovaries. ResultAs compared with the blank group， large follicles with polycystic expansion were found in the ovaries of the model group， no dominant follicles were found， the granular layer of follicles decreased and arranged loosely， and the number of corpus luteum decreased significantly. Serum T， LH， AMH and LH/FSH increased in the model group （P<0.05， P<0.01）， while FSH and E₂ decreased （P<0.05， P<0.01）. The relative protein expression levels of NALP3， ASC， Caspase-1， NF-κB， IL-1β， IL-18， and IL-6 increased （P<0.05， P<0.01） in the ovaries of the model group. Compared with the model group， the low， medium， and high-dose Jianpi Yishen Huazhuo prescription groups and the metformin group showed growing follicles and corpus luteum at all levels， the number of cystic expanding follicles decreased， the thickness of follicular granular layer increased， the number of follicular fluid increased， mature follicles were visible， and the local morphology of oocytes was complete. Serum T， LH， AMH， and LH/FSH in these groups decreased （P<0.05， P<0.01）， while E₂ and FSH increased （P<0.05）. The relative protein expressions of NALP3， ASC， Caspase-1， NF-κB， IL-1β， IL-18， and IL-6 in the ovaries of these groups decreased （P<0.05， P<0.01）. There was no significant difference among the treatment groups. ConclusionBy inhibiting the activation of NLRP3 inflammasome， Jianpi Yishen Huazhuo prescription reduces the release of NALP3， ASC， Caspase-1， NF-κB， IL-18， IL-1β， and IL-6 inflammatory factors in ovarian tissues， regulates endocrine level， and effectively reduces PCOS inflammatory statu， so as to play a role in improving ovarian function.

ObjectiveTo investigate the material basis and mechanism of Arnebia euchroma against hepatocarcinoma by network pharmacology， and to verify the potential targets of A. euchroma against hepatocarcinoma by molecular docking and experiments. MethodThe main active ingredients of A. euchroma were collected by retrieving the literature through China National Knowledge Network （CNKI） and Traditional Chinese Medicine System Pharmacology Database and Analysis Platform（TCMSP）. The active ingredients were screened out by FAFDrug4 platform according to the pharmacokinetics （ADME） properties of the drugs. The screening compounds and liver cancer targets were collected by using several databases and analyzed by drawing Venn diagrams. The protein-protein interaction （PPI） network was constructed by Cytoscape and STRING database. DAVID database was used to perform Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis of key targets. Autodock was used to perform molecular docking of targets on core pathways. Cell counting kit-8 （CCK-8） experiment was carried out to validate the activities of five naphthoquinones. Based on the predicted results in the H22 tumor-bearing mouse model， the key targets of isovalerylshikonin against hepatocarcinoma were verified by hematoxylin-eosin （HE） staining， enzyme-linked immunosorbent assay （ELISA）， and Western blot assay. ResultFifty-five active ingredients and 34 targets of active ingredients against hepatocarcinoma were screened out. The active molecules with high degree values in the “drug-active ingredient-target-disease” network were mainly naphthoquinones. PPI network obtained several core targets of A. euchroma against hepatocarcinoma. Twenty-two pathways were screened out by KEGG analysis， mainly involving phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt）， vascular endothelial growth factor （VEGF）， tumor necrosis factor （TNF）， and other signaling pathways. The results of molecular docking showed that the five naphthoquinones had a good affinity with the targets of the PI3K/Akt signaling pathway. The results of CCK-8 and animal experiments showed that the lipid-soluble component isovalerylshikonin had good anti-cancer potential， and the high-dose group reduced the serum levels of VEGF and alpha fetoprotein （AFP） levels and elevated interferon-γ （IFN-γ） level （P<0.05， P<0.01）. The high-dose group also down-regulated phosphorylate（p）-Akt （Ser473） and B-cell lymphoma （Bcl）-2 protein expressions and up-regulated Bcl-2-antagonist of cell death （Bad） protein expression （P<0.01）. ConclusionA. euchroma can inhibit hepatocarcinoma cell proliferation and tumor angiogenesis and induce cancer cell apoptosis through the PI3K/Akt signaling pathway， which provides ideas and clues for the subsequent in-depth investigation of its specific mechanism.