OBJECTIVES: To explore the correlation of ELFN1 expression level with prognosis of colorectal cancer and its regulatory role in colorectal cancer cell proliferation and metastasis. METHODS: We analyzed the expression levels of ELFN1 across 33 cancer types using publicly available databases and identified differential genes related to ELFN1 in colorectal cancer. Gene function annotation and enrichment analysis were used to identify the involved signaling pathways. Logistic analysis, Kaplan-Meier analysis and Cox regression analysis were performed to evaluate the correlation between ELFN1 expression and clinicopathological parameters and survival of colorectal cancer patients. qPCR and Western blotting were used to validate the expression levels of ELFN1 in different colorectal cancer cell lines and tissues, and Transwell and EDU experiments were carried out to assess the effect of ELFN1 knockdown on biological behaviors of SW480 cells. RESULTS: ELFN1 was highly expressed in 14 cancers, and its expression was significantly higher in colon cancer tissues than in adjacent tissues. A high expression of ELFN1 mRNA was associated with a poorer overall survival of colorectal cancer patients. Cox regression analysis indicated that ELFN1 expression was an independent prognostic factor for overall survival of the patients. ELFN1 was significantly enriched in tumor metastasis and proliferation and participated in several tumor signaling pathways. The colon cancer cell lines showed significantly higher expression levels of ELFN1 than normal cells, ELFN1 knockdown obviously inhibited proliferation and migration of SW480 cells in vitro. CONCLUSIONS ELFN1 is overexpressed in colorectal cancer and is associated with poor clinical prognosis of the patients. A high ELFN1 expression is associated with malignant phenotypes of colorectal cancer and promotes cancer cell proliferation and metastasis, suggesting its potential as a prognostic biomarker for colorectal cancer.
Humans ; Colorectal Neoplasms/diagnosis* ; Cell Proliferation ; Prognosis ; Cell Line, Tumor ; Biomarkers, Tumor/metabolism* ; Neoplasm Metastasis ; Gene Expression Regulation, Neoplastic ; Nerve Tissue Proteins/metabolism* ; Female ; Male

BACKGROUND:As a degradable scaffold material for bone tissue engineering,lactic acid is widely used in tissue regeneration and repair research,and plays an important role in promoting tissue healing,new bone formation and angiogenesis. OBJECTIVE:To observe the effect of lactic acid degradation products on osteoclasts and to investigate the effects of lactic-interfered osteoclast conditioned medium on the proliferation,migration and tube-forming capacity of human umbilical vein endothelial cells. METHODS:(1)The mouse monocyte macrophage cell line RAW264.7 at logarithmic growth period was selected,and adherent cells were cultured in the osteoclast induction medium(DMEM medium with nuclear factor-κB receptor-activating factor ligand and 10%fetal bovine serum)containing different concentrations of lactic acid(0,5,10,20 mmol/L).After 5 days of culture,tartrate-resistant acid phosphatase staining and cytoskeletal fibrillar actin staining were conducted.After 24 hours of culture,RT-PCR was used to detect the mRNA expression of tartrate-resistant acid phosphatase 5.(2)RAW264.7 cells at logarithmic growth period were selected and adherent cells were divided into two groups.Control group was cultured in the osteoclast induction medium,while experimental group was cultured in the osteoclast induction medium containing 10 mmol/L lactic acid.After 5 days of culture,the medium in each group was removed and the cells in the two groups were cultured in the serum-free DMEM medium for another 24 hours.Cell supernatant was then collected and used as the conditioned medium after mixed with an equal volume of DMEM medium containing 10%fetal bovine serum.Human umbilical vein endothelial cells at the logarithmic growth phase were taken and separately co-cultured with the conditioned medium of the control and experimental groups.The proliferation,migration and tube-forming ability of human umbilical vein endothelial cells were observed by cell counting kit-8 assay,migration assay,scratch assay and tube-forming assay.The mRNA and protein expression of angiogenesis-related genes and proteins were observed by RT-PCR and western blot. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining and cytoskeletal fibrillar actin staining showed that 5 and 10 mmol/L lactic acid promoted osteoclastic differentiation of RAW264.7 cells and the promoting effect of 10 mmol/L lactate was more significant.RT-PCR results showed that the expression of tartrate-resistant acid phosphatase-5 mRNA of osteoclast-related genes was the highest when the lactic acid concentration was 5,10,and 20 mmol/L(P<0.05),especially 10 mmol/L.Compared with the control group,the proliferation,migration and tube-forming abilities of human umbilical vein endothelial cells were significantly increased in the experimental group(P<0.05).Compared with the control group,the expression levels of vascular endothelial growth factor and angiogenin 1 mRNA and protein were increased in the experimental group(P<0.05).To conclude,lactate-induced osteoclast conditioned medium could promote the angiogenesis of endothelial cells,and the mechanism may be related to the promotion of the expression of vascular endothelial growth factor and angiogenin 1.

BackgroundDepression is a prevalent emotional problem in adolescents， and parental psychological control is an important predictor of adolescent depression. However， existing research on the acting mechanism between the two is not adequate. ObjectiveTo explore the pathway of negative perfectionism and academic stress between parental psychological control and depressive symptoms among secondary school students， so as to provide references for reducing the incidence risk of depression in such population. MethodsFrom February to April 2023， 1 100 students across 2 middle schools and 2 high schools in Zhongshan city were selected as subjects. The survey was conducted adopting Parental Psychological Control Questionnaire， Chinese Frost Multidimensional Perfectionism Scale （CFMPS）， sense of academic stress subscale in Mental Health Inventory of Middle School Student （MMHI-60） and Center for Epidemiological Studies-Depression Scale （CES-D）. Spearman correlation analysis was adopted to examine the correlation between scores of all scales above， and Amos 24.0 was used to test the mediating path of negative perfectionism and academic stress between parental psychological control and depressive symptoms among secondary school students. ResultsAmong the 1 009 valid questionnaires withdrew （91.73% of the total）， 261 students were detected to have depressive symptoms （25.87%）. As the results of Spearman correlation analysis showed， the scores of the Parental Psychological Control Questionnaire， score of negative perfectionism dimension in CFMPS， score of sense of academic stress subscale in MMHI-60 and CES-D score were positively correlated with each other （r=0.323~0.644， P<0.05 or 0.01）. The direct effect value of parental psychological control on depressive symptoms in secondary school students was 0.128 （95% CI： 0.061~0.201）， accounting for 31.37% of the total effect. Negative perfectionism and academic stress played independently as intermediatory roles between parental psychological control and depressive symptoms in secondary school students， and the indirect effect values were 0.099 （95% CI： 0.068~0.133） and 0.100 （95% CI： 0.060~0.143）， accounting for 24.27% and 24.51% of the total effect， respectively. Negative perfectionism and academic stress acted combinedly as the chain effect pathway between parental psychological control and depressve symptoms in secondary school students， with the indirect effect value of 0.081 （95% CI： 0.060~0.106） accounting for 19.85% of the total effect. ConclusionParental psychological control can affect the depressive symptoms among secondary school students directly， and through independent or chain paths of negative perfectionism and academic stress indirectly. ［Funded by Zhongshan Social Welfare Technology Research Project （number， 2022B1060）］

Objective To investigate the influence of different embryo development rates on the clinical outcomes of assisted reproductive technology(ART).Methods The clinical data of female patients who underwent in vitro fertilization and intracytoplasmic sperm injection embryo transfer in Reproductive Medicine Center of The First Affiliated Hospital of Naval Medical University from Jan.1,2015 to Dec.31,2023 were retrospectively analyzed.A total of 1 556 cycles were included.Group A was transferred on day 3,and they were assigned to subgroups according to the embryo development rates until day 3:subgroup A1(≤6 cell stages),subgroup A2(7-9 cell cleavage stages),or subgroup A3(≥10 cell cleavage stages).Group B was transferred on day 4,and they were assigned to subgroups according to the embryo development rates until day 4:subgroup B1(cleavage stages),subgroup B2(1st or 2nd period blastocyst or morula stages),or subgroup B3(3rd period blastocyst or higher stages).Group C was transferred on day 5,and they were assigned to subgroups according to the embryo development rates until day 5:subgroup C1(1st or 2nd period blastocyst or morula stages),subgroup C2(4th or 5th period blastocyst stages),or subgroup C3(6th period blastocyst stages).Group D was transferred on day 6,and they were assigned to subgroups according to the embryo development rates until day 6:subgroup D1(morula or 1st or 2nd period blastocyst stages),or subgroup D2(5th or 6th period blastocyst stages).The clinical pregnancy and live birth rates were calculated for each group.Results Pairwise comparisons of the subgroups A1,B1,C1 and D1,all with relatively slow development rates,showed that the clinical pregnancy rates were 23.7%,37.3%,26.9%and 35.9%,respectively(P＜0.05),the live birth rates were 16.4%,28.4%,19.2%and 26.9%,respectively(P＜0.05),and the clinical pregnancy rate and live birth rate were both the highest in the B1 group.Pairwise comparisons of the subgroups A2,B2,C2 and D2 with normal development rates,the clinical pregnancy rates were 58.0%,59.4%,62.2%and 61.5%,respectively(P＜0.05),the live birth rates were 47.5%,49.4%,53.8%and 52.3%,respectively(P＜0.05),and the clinical pregnancy rate and live birth rate were both the highest in the subgroup C2.Pairwise comparisons of the subgroups A3,B3 and C3 with relatively fast development rates showed that the clinical pregnancy rates were 62.2%,64.6%and 63.5%,respectively(P＜0.05),the live birth rates were 52.2%,56.9%and 54.1%,respectively(P＜0.05),and the clinical pregnancy rate and live birth rate were both the highest in the subgroup B3.Conclusion The 4th day fast-developing blastocysts have better development potential and clinical outcomes.Embryos with slower development rate should be transferred on the 4th day,and embryos with normal development rate are recommended to be cultured and transferred to the 5th day.

Objective:To investigate improvement effect of osthol(OST)on inflammatory response in gestational diabetes mel-litus(GDM)rats by regulating cGAS-STING signaling pathway.Methods:GDM rat model was established by intraperitoneal injection of 35 mg/kg streptozotocin on the fifth day of pregnancy,and randomly grouped into GDM group,OST-L group(1 mg/kg),OST-M group(10 mg/kg),OST-H group(50 mg/kg),OST-H+DMXAA group(50 mg/kg OST solution+25 mg/kg DMXAA),positive control group(200 mg/kg metformin hydrochloride),with 10 rats in each group,another 10 rats with 5-day pregnancy were regarded as con-trol group.After intervention,glucose meter was applied to detect fasting blood sugar;ELISA was applied to detect insulin and TNF-α,IL-6,IL-1β levels;automatic biochemical analyzer was applied to detect contents of total cholesterol(TC),triglyceride(TG)and high-density lipoprotein(HDL);pancreatic tissues were separated and Western blot was applied to detect cGAS and STING proteins expressions in pancreatic tissue;HE staining was applied to detect pathological changes in pancreatic tissue.Results:Compared with control group,pathological damage in GDM group was severe,cGAS and STING protein expressions,pathological score,TNF-α,IL-6,IL-1β,TC,TG,insulin and fasting blood sugar levels were increased obviously,HDL level was decreased obviously(P<0.05);compared with GDM group,pathological damage in OST-L group,OST-M group and OST-H group was improved,cGAS and STING protein expressions,pathological score,TNF-α,IL-6,IL-1β,TC,TG,insulin and fasting blood sugar levels were decreased obviously,HDL level was increased obviously(P<0.05);compared with OST-H group,pathological damage in OST-H+DMXAA group was aggravated,cGAS and STING protein expressions,pathological score,TNF-α,IL-6,IL-1β,TC,TG,insulin and fasting blood sugar levels were increased obviously,HDL level was decreased obviously(P<0.05),however,there was no difference in posi-tive control group(P>0.05).Conclusion:OST can alleviate inflammatory response in GDM rats by inhibiting cGAS-STING signaling pathway.

Objective To investigate the expression of Cyclin O(CCNO)in cervical cancer and its molecular mechanism in the progression of cervical cancer.Methods The pathological sections of 60 patients with cervical cancer in the Department of Oncology and Gynecology of the First Affiliated Hospital of Bengbu Medical University from January 2018 to December 2023 were collected,and the expression of CCNO in cervical cancer was detected by immunohistochemistry.The cells were divided into Vector group,CCNO group,siNC group and siCCNO group by transfection technology.CCK-8 assay and colony formation assay were used to evaluate the ability of CCNO to affect cell proliferation.Transwell assay and wound healing assay were used to evaluate the effect of CCNO on cell invasion and migration.Glycolysis assay and Western blot assay were used to evaluate the effect and mechanism of CCNO on glycolysis of cervical cancer cells.Results The results of immunohistochemistry and WB showed that CCNO was highly expressed in cervical cancer(P<0.001).Overexpression of CCNO promoted the proliferation,invasion,migration and glycolysis of cervical cancer cells(P<0.05),whereas the opposite effect inhibited the proliferation,invasion,migration and glycolysis of cervical cancer cells(P<0.05).Bioinformatics analysis and WB results showed that CCNO overexpression may regulate the occurrence of cervical cancer by activating the PI3K/AKT signaling pathway(P<0.05).Conclusion Cyclin O may promote the proliferation and metastasis of cervical cancer cells by regulating glycolysis.

Objective To screen and analyze the differentially expressed genes (DEGs) related to oxidative stress in a silicosis mouse model using transcriptome sequencing technology. Methods i) A total of 30 workers without occupational dust-exposed history were selected as the control group and 17 patients with silicosis were selected as the silicosis group using a judgment sampling method. The levels of glutathione and malondialdehyde in the plasma of workers in the two groups were determined by enzyme-linked immunosorbent assay. ii) RAW264.7 cells in the logarithmic growth phase were randomly divided into the control group and the silica group, treated with 0 and 50 mg/L silica suspensions for 24 hours. Protein expression of superoxide dismutase 2 (SOD2), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in the cells was determined by Western blotting. iii) The specific pathogen free male C57BL/6 mice were randomly divided into the control group and the silicosis model group, with 10 mice in each group. Mice were exposed to 50 μL of 0.9% sodium chloride solution and silica suspension at a mass concentration of 100 g/L, respectively, using a single tracheal exposure method. After 28 days of exposure, the pathological changes of mouse lung tissues were observed. Transcriptome sequencing was used to screen DEGs in the lung tissues of the silicosis mouse model, and gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed. The expression of DEGs was verified using quantitative real-time polymerase chain reaction (qPCR). Results i) The level of malondialdehyde in the patients′ plasma was higher (P<0.01), while the level of glutathione was lower (P<0.01) in the silicosis group than that of the control group. ii) The relative expression of SOD2 protein decreased (P<0.05), while the relative expression of IL-6 and TNF-α proteins increased (all P<0.05) in the silica group of RAW264.7 cells compared with the control group. iii) The pathological results of lung tissues showed that the alveolar structure of mice was destroyed and silicotic nodules were formed in the silicosis model group. Transcriptome sequencing identified 3 703 DEGs, of which 3 199 were significantly down-regulated and 504 were significantly up-regulated. The GO enrichment analysis results showed that the DEGs were significantly enriched in biological processes such as oxidative stress, inflammation, immunity and hypoxia, with cellular components mainly located in membranes, cytoplasm, and nucleus. Molecular functions were enriched in oxidoreductase activity, protein binding, and adenosine triphosphate binding. The KEGG enrichment analysis results showed that the DEGs were mainly involved in the phosphatidylinositol 3-kinase-protein kinase B signaling pathway, cyclic adenosine monophosphate signaling pathway, chemokine signaling pathway, and apoptosis signaling pathway. A total of 28 DEGs involved in the "oxidative stress response" pathway were screened by GO enrichment analysis. The qPCR verification results showed that the relative expression of DEGs carbonic anhydrase 3 (Car3), matrix metalloproteinase 9 (Mmp9), and MutY DNA glycosylase (Mutyh) involved in the "oxidative stress response" of lung tissues in the silicosis model group were lower than those of the control group (all P<0.05). Conclusion Oxidative stress response exists in silicosis patients. The oxidative stress-related genes Car3, Mmp9, and Mutyh are altered in the mouse lung tissues of the silicosis model through the oxidative stress pathway, suggesting that they could be new targets for the treatment of silicosis.

OBJECTIVE To investigate the inhibitory effect mechanism of Anzheng Kangliu Decoction against colorectal cancer by suppressing glycolysis through regulation of the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)signaling pathway.METHODS Human colorectal cancer SW620 cells were treated with Anzheng Kangliu De-coction,and cell proliferation and migration abilities were assessed.Forty-two BALB/c nude mice were randomly divided into a blank control group,model group,5-fluorouracil(5-FU)group(0.025 g·kg-1),Anzheng Kangliu Decoction low-dose group(7.67 g·kg-1),medium-dose group(15.34 g·kg-1),and high-dose group(30.68 g·kg-1).The inhibitory effect of Anzheng Kan-gliu Decoction on subcutaneous xenograft tumors was evaluated by observing body weight,tumor volume,tumor mass,HE staining,im-munohistochemical staining of Ki67 and other indicators.High-throughput transcriptome sequencing was performed to identify differen-tially expressed genes and pathways in tumor tissues between the model group and the Anzheng Kangliu Decoction medium-dose group,elucidating the potential mechanism of Anzheng Kangliu Decoction against colorectal cancer.Glucose and lactate assay kits were used to measure glucose consumption and lactate production in SW620 cells and tumor tissues after Anzheng Kangliu Decoction intervention.Western blot was employed to detect the expression levels of p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR,hexokinase 2(HK2),and lactate dehydrogenase A(LDHA)in SW620 cells and tumor tissues following Anzheng Kangliu Decoction treatment.RE-SULTS In vitro cell experiments demonstrated that,compared with the blank control group,Anzheng Kangliu Decoction intervention significantly inhibited the proliferation and migration of SW620 cells(P<0.01),reduced glucose consumption and lactate production(P<0.05,P<0.01),and downregulated the protein expression levels of p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR,HK2,and LDHA(P<0.05,P<0.01).In vivo animal experiments revealed that,compared with the model group,Anzheng Kangliu Decoction suppressed the growth of subcutaneous xenograft tumors in nude mice(P<0.01),increased tumor tissue necrosis,decreased glucose consumption and lactate production in tumor tissues(P<0.05,P<0.01),and reduced the protein expression levels of p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR,HK2,and LDHA(P<0.05,P<0.01).CONCLUSION Anzheng Kangliu Decoction exerts an in-hibitory effect on colorectal cancer,and its mechanism may be associated with the suppression of glycolysis through regulation of the PI3K/Akt/mTOR signaling pathway.

Objective:To explore the short-term clinical effects of deep cervical lymph node-venous anastomosis in the treatment of Alzheimer’s disease (AD).Methods:A prospective exploratory study was conducted on the treatment of AD patients using the cervical deep lymph node-venous anastomosis technique in Scar and Wound Treatment Department, Plastic Surgery Hospital, Chinese Academy of Medical Sciences, from September to October 2024. The patients underwent high-frequency ultrasound to locate deep cervical lymph nodes and the external jugular vein. Under general anesthesia, bilateral deep cervical lymph node-venous anastomoses were performed. Indocyanine green (ICG) lymphography was conducted via subcutaneous injection behind the ear to visualize lymph nodes in levels Ⅱ and Ⅲ. After making a skin incision along the posterior margin of the sternocleidomastoid muscle, the external jugular vein, internal jugular veins, and associated lymph nodes were exposed. Adjacent veins were selected for anastomosis of lymph node. Using microsurgical techniques, end-to-side or end-to-end anastomosis was completed for lymph nodes in levels Ⅱ and Ⅲ. Preoperative assessments included the mini-mental state examination (MMSE, a higher score indicates better cognitive function), Alzheimer’s disease assessment scale-cognitive subscale (ADAS-Cog, a higher score indicates greater impairment of cognitive function), Alzheimer’s disease cooperative study scale for activities of daily living (ADCS-ADL, a higher score indicates better ability to perform daily activity), and neuropsychiatric inventory (NPI, a higher score indicates more severe behavioral and emotional symptom). Postoperative follow-up included the same scales to observe changes in cognitive function, activities of daily living, and emotional communication.Results:Four patients (1 male, 3 females, aged 58-79 years) with AD were included. All were diagnosed based on cerebrospinal fluid biomarkers. All patients successfully underwent bilateral deep cervical lymph node-venous anastomoses. On average, 4.3 (2-7 per person) anastomoses were performed per patient. Surgical procedures lasted an average of 6.5 h (5.5-8.5 h) with minimal blood loss (less than 50 ml). Patients resumed normal activity within 6 hours postoperatively and were discharged after an average of 4.1 d (3.5-5.0 d). Postoperative complications included one case each of aspiration pneumonia, lower limb venous thrombosis, and transient delirium, all of whom resolved without long-term effects. Clinical symptoms, including memory decline, mood swings, and anxiety, showed varying degrees of improvement. Patients reported enhanced quality of life, emotional stability, and social engagement, confirming the procedure’s safety and potential cognitive benefits. At one month postoperatively, the MMSE scores of the four patients increased by an average of 0.8 points compared to preoperative levels. Additionally, the two patients who completed the ADAS-Cog assessments showed a decrease in their scores (reduced by 1.0 points and 11.3 points, respectively, compared to preoperative scores), indicating a certain degree of improvement in cognitive function during this period. The ADCS-ADL and NPI scores of four patients varied significantly, without showing any clear pattern.Conclusion:Lymphovenous anastomosis of the deep cervical lymph node-venous anastomosis may provide a new surgical intervention approach for AD, but further large-scale studies and long-term follow-up are needed to validate its safety and effectiveness.

This paper reported a type 1 diabetes patient who had severe diabetic nephropathy, retinopathy, hypertension, and hypothyroidism before pregnancy. The patient's blood glucose control was poor before pregnancy, and the complications were not properly treated. This was an unintended pregnancy, with a pre-pregnancy glycated hemoglobin A1c of 7.8% and early pregnancy urine protein of 3.81-4.53 g/24 h. Considering the patient's poor blood glucose control before pregnancy and the lack of proper treatment for multiple complications including nephropathy, a multidisciplinary consultation at an external hospital recommended termination of the pregnancy. However, the patient was determined to continue the pregnancy and was referred to Peking University First Hospital. Through strict blood glucose control, monitoring and evaluation of complications, and comprehensive management, the patient's blood glucose and blood pressure were well controlled during pregnancy. Regular monitoring of urine protein, renal function, and ocular fundus was conducted. At 31 weeks and 4 days of gestation, the patient's 24-hour urine protein significantly increased. After promoting fetal lung maturity, a cesarean section was performed at 34 weeks and 1 day of gestation, resulting in a successful delivery with good maternal and neonatal outcomes. At the 42-day postpartum follow-up, the patient's blood glucose and blood pressure were stable, urine protein returned to pre-pregnancy levels, and the infant was in good general condition.