abstract:
Objective To construct triggering receptor expressed on myeloid cells 2(TREM2) gene knockout(TREM2-/-) mice using CRISPR/Cas9 technology, to breed TREM2-/- mice and to analyze the genotype of TREM2-/- mice. Methods CRISPR/Cas9 technology was used to selectively knock out exon 2-3 regions of TREM2 gene to construct a TREM2-/- mouse model, and the genetic background of all mice was C57BL/6J. Polymerase chain reaction(PCR) was used to identify the genotype of mice. Quantitative real-time PCR(qPCR) and Western blot were used to detect the expression level of TREM2 in major tissues of mice, and the authenticity and scientific nature of PCR identification results were verified from mRNA level and protein level. According to the sgRNA sequence, the possible off-target sites were predicted on the CCTop website, and the tail DNA of mice was extracted and amplified by PCR and then Sanger sequencing was performed to detect whether there was off-target effect in TREM2-/- mice. Results TREM2-/- mice were successfully constructed by CRISPR/Cas9 technology, and the mice were genotyped. The results of agarose gel electrophoresis showed that the mouse genotype with only 415 bp band amplified was wild type(WT), the mouse genotype of the 449 bp band amplified only was TREM2-/-, and the mouse genotypes amplified with 415 bp and 449 bp double bands were heterozygous. qPCR results showed that compared with WT mice, the mRNA expression of TREM2 in heart and brain tissues of TREM2-/- mice was down-regulated(P-/- mice was reduced(P-/- mice. Conclusion TREM2-/- mice are successfully constructed and bred, a reliable genotype identification method is established, the genetic stability of the mouse model is verified, which will provide an important genetic animal model for the study of TREM2 gene function.

abstract:
Objective To explore the effect of dendrobium officinale polysaccharides(DOP) on necrotizing enterocolitis(NEC) in neonatal mice and to preliminarily explore the potential molecular mechanisms. Methods Seven-day-old C57BL/6J mice were randomly divided into four groups: control(CTRL) group, necrotizing enterocolitis(NEC) group, low-dose DOP treatment(DOPL+NEC) group and high-dose DOP treatment(DOPH+NEC) group. The NEC model was established by hypoxia, cold stimulation, hypertonic feeding and intraperitoneal injection of lipopolysaccharide(LPS). At the end of the experiment, the small intestine tissues were collected. The general condition of the mice was observed, and body weight was recorded. Hematoxylin-eosin(HE) staining was used to observe pathological changes in the small intestine tissues. The expression levels of E-cadherin, Occludin and Claudin-1 were detected by immunohistochemistry and Western blot. The concentrations of tumor necrosis factor-α(TNF-α), interleukin(IL)-1β, IL-6 and IL-10 were measured by ELISA. The levels of lactate dehydrogenase(LDH), malondialdehyde(MDA), superoxide dismutase(SOD) and glutathione(GSH) were detected using commercial kits. The expression levels of nuclear factor erythroid 2-related factor 2(Nrf2) and heme oxygenase-1(HO-1) proteins were detected by immunohistochemistry and Western blot. Results Compared with the CTRL group, the NEC group exhibited decreased body weight and increased HE pathological scores(P

abstract:
Objective To explore the expression profile of core 1 β1,3-galactosyltransferase 1(C1GALT1) in glioblastoma(GBM) and to elucidate its impact on the initiation and progression of GBM. Methods The expression levels and prognostic significance of C1GALT1 in GBM were analyzed using the GEPIA and CGGA databases. Two representative glioblastoma cells(U251 and LN18) were selected to construct C1GALT1-knockdown cell lines and performed in vitro experiments. The Cell Counting Kit-8(CCK-8) and Transwell assays were employed to evaluate the impact of C1GALT1 on proliferation, migration and invasion of GBM cells. Transcriptome data were analyzed to identify potential signaling pathways. Senescence β-Galactosidase Staining Kit was used to detect β-galactosidase activity. Results Analysis of GEPIA and CGGA databases revealed that C1GALT1 was significantly upregulated in GBM tissues compared to adjacent non-cancerous tissues(P

abstract:
Objective To investigate the effect and mechanism of silencing circFADS2 on ferroptosis in colorectal cancer(CRC) cells. Methods The human colon adenocarcinoma cell line SW480 was used as the research object. circFADS2 siRNA interference plasmid(si-circFADS2) and its negative control si-NC, miR-152-3p inhibitor and its negative control inhibitor-NC, miR-152-3p mimics and its negative control mimics NC, SLC7A11 overexpression plasmid(oe-SLC7A11) and its negative control vector were transfected into SW480 cells. Methylthiazolyldiphenyl-tetrazolium bromide(MTT) method was used to detect cell proliferation activity; The contents of ferrous ion(Fe2+), malondialdehyde(MDA) and the activities of superoxide dismutase(SOD) and glutathione(GSH) in cells were detected by chemical method. Dichlorodihydrofluorescein diacetate(DCFH-DA) probe was used to detect the level of reactive oxygen species(ROS) in cells. Quantitative Real-time polymerase chain reaction(qPCR) was used to detect the expression levels of miR-152-3p and SLC7A11 mRNA in cells. Western blot was used to detect the expression levels of ferroptosis-related proteins such as SLC7A11 and GPX4 in cells. Dual luciferase reporter gene experiment was used to verify the sponge adsorption relationship between miR-152-3p and circFADS2, and the targeted regulation relationship between miR-152-3p and SLC7A11. Results Compared with blank group, the proliferation activity of si-circFADS2 group decreased, the levels of Fe2+, MDA and ROS increased, and the levels of SOD and GSH decreased; At the same time, the expression level of miR-152-3p increased, and the protein expression levels of SLC7A11 and GPX4 decreased in cells(all P2+, MDA and ROS in cells decreased, and the levels of SOD and GSH increased; At the same time, the expression level of miR-152-3p decreased, and the protein expression levels of SLC7A11 and GPX4 increased(all P

abstract:
Objective To investigate the effect of polystyrene microplastics(PS-MPs) on myocardial injury in mice and its molecular mechanism. Methods A total of 60 male C57BL/6 mice were randomly divided into normal saline group, 0.1, 1, 10 mg/kg PS-MPs exposed group, and doxorubicin [5 mg/(kg·w)] group treated for 8 weeks. After treatment, we measured blood pressure, cardiac organ coefficient, cardiac histopathological changes, oxidative stress markers reactive oxygen species(ROS), malondialdehyde(MDA), glutathione(GSH), 4-hydroxynonenal(4-HNE) and nuclear factor E2-related factor 2(Nrf2), serum centroid injury markers creatine kinase MB(CK-MB) and troponin(cTnT), ferroptosis marker recombinant glutathione peroxidase 4(GPX4), Recombinant solute carrier family 7(SLC7A11) as well as ferrous ions(Fe2+). Results Compared with the negative control group, vacuolation, inflammatory infiltration and collagen fiber deposition were evident in the hearts of mice after PS exposure. The levels of myocardial injury markers CK-MB and cTnT significantly increased. Cardiac organ coefficient decreased, blood pressure increased, oxidative stress markers and ferroptosis markers increased. Conclusion PS-MPs exposure can induce oxidative stress and activate ferroptosis pathway, resulting in myocardial injury in mice.

abstract:
Objective To construct a humanized mouse model of the interleukin-9 receptor(IL-9R) coding DNA sequence(CDS) gene and to verify the genotype and IL-9R expression in mice. Methods The CRISPR/Cas9 genome editing technology was used to replace the exon 2-7 fragment of the il-9r gene in mouse embryonic stem cells with the corresponding human IL-9R sequence. After verifying the completion of the gene fragment replacement, tetraploid embryos were constructed and microinjected back into the oviducts of surrogate mice. Through surrogacy by female mice, homozygous humanized mice were obtained. DNA was extracted from the homozygous humanized mice IL-9R CDS gene, and their genotypes were identified by agarose gel electrophoresis after PCR amplification. Western blot was used to detect the expression of IL-9R in the spleen and thymus of homozygous humanized mice with either wild-type(WT) or IL-9R gene humanization. Results Gel electrophoresis after PCR amplification showed that mice with only a 1 805 bp band amplified using WT primers were wild-type, while mice with 2 553 bp and 2 340 bp bands amplified using 5KI and 3KI primers, respectively, were homozygous humanized mice with IL-9R CDS gene. Western blot results indicated that the tissues of homozygous humanized mice model with IL-9R CDS gene expressed IL-9R significantly. Conclusion The humanized mouse model with IL-9R CDS gene has been successfully constructed and characterized.

abstract:
Objective To investigate the mechanism of miR-224-5p on proliferation, apoptosis, invasion and migration of human hepatocellular carcinoma HepG2 cells. Methods The RNA expression levels of miR-224-5p and early growth responsive gene 2(EGR2) in patients with hepatocellular carcinoma were obtained from the TCGA dataset. Normal human hepatocytes LO2 and hepatoma cells HepG2 were cultured in vitro, and the HepG2 cells were transfected with lentiviral vectors(knockdown of miR-224-5p), small interfering RNA fragments or overexpression vectors(interference and overexpression of EGR2). The expression levels of miR-224-5p and EGR2 in hepatocellular carcinoma cDNA chips and cells were detected by quantitative real-time PCR(qPCR). The expression level of EGR2 protein was detected by Western blot. Dual luciferase reporter gene assay was used to detect the binding of miR-224-5p to EGR2. HepG2 cells positive rate were detected by EdU assay, apoptosis rate was detected by flow cytometry, cell invasion number was detected by Transwell assay, and cell mobility was detected by scratch assay. Results Compared with paracancerous tissues, the expression of miR-224-5p was increased and the expression of EGR2 mRNA decreased in HCC tissues. Compared with LO2 group, the expression of miR-224-5p in HepG2 cells increased, and the expression of EGR2 mRNA and protein decreased. Compared with the Lv-sh-NC group, the 24 h EdU positive cell rate, cell invasion number, and 48 h cell mobility of HepG2 cells in the Lv-sh-miR-224-5p group decreased, while the apoptosis rate increased. Compared with Oe-NC group, 24 h EdU positive cell rate, cell invasion number, and 48 h cell mobility of HepG2 cells in Oe-EGR2 group decreased, while apoptosis rate increased. Compared with Lv-sh-NC group, the expression of EGR2 protein in Lv-sh-miR-224-5p group increased. Compared with Lv-sh-miR-224-5p+si-NC group, 24 h EdU positive cell rate, cell invasion rate, and 48 h cell mobility of HepG2 cells in Lv-sh-miR-224-5p+si-EGR2 group increased, while apoptosis number decreased. Conclusion miR-224-5p can promote proliferation, invasion, and migration of HepG2 cells and inhibit apoptosis via binding with EGR2.

abstract:
Objective To investigate the method and evaluation of using different doses of vancomycin(VCM) to construct an acute kidney injury(AKI) model in SD rats. Methods The 6-week-old male SD rats were randomly divided into 4 groups: normal control(NC) group, VCM(200, 400, 600) mg/kg group. The drug was given by intraperitoneal injection once a day for 7 days. The levels of blood urea nitrogen(BUN) and creatinine(Cre) were detected. The kidney was weighed and the kidney body ratio was calculated. Renal tissue was stained with HE and PAS, and the pathological injury of kidney was observed under light microscope. Results With the increase of VCM dose, serum BUN and Cre levels in each dose group showed an upward trend(F=3.85,P<0.05;F=9.02,P<0.01). Compared with the VCM(200, 400) mg/kg groups, the Cre level in the VCM 600 mg/kg group was the highest, with statistically significant differences(P<0.01,P<0.05). As the VCM dose increased, body weight of rats in each dose group decreased(F=54.81,P<0.01), while total kidney weight and kidney body ratio increased(F=11.13,F=14.43, bothP<0.01). Significant statistically differences(P<0.01,P<0.05)were observed in body weight, total kidney weight and kidney body ratio between the VCM 600 mg/kg group and the VCM(200, 400) mg/kg groups. HE staining and PAS staining showed that renal tubules in each dose group were damaged to varying degrees. With the increase of VCM dose, the pathological score of kidney injury increased(F=105.80,F=28.89, bothP<0.01), and the pathological damage of renal tubular epithelial cells such as shedding, swelling and cast formation was the most serious in VCM 600 mg/kg group. Conclusion The AKI model in rats can be stably established by intraperitoneal injection of VCM at 600 mg/kg once a day for 7 days.

abstract:
Objective To investigate the effect and mechanism of M2 macrophage polarization induced by HCT116 with highly metastatic colorectal cancer cells on immune escape in colorectal cancer. Methods After the co-culture of colorectal cancer cells conditioned medium(CM) and PMA-induced M0 macrophages, the polarization of M2 macrophages was observed by flow cytometry, real-time polymerase chain reaction(qPCR) and enzyme-linked immunosorbent assay(ELISA) experiments. The CMM0, CMSW480-Mφ and CMHCT116-Mφ were co-cultured with HCT116 cells, and the expression of programmed death-ligand 1(PD-L1) was detected by Western blot and qPCR. At the same time, the stimulated HCT116 cells were co-cultured with human T lymphocytes to detect the survival of HCT116 cells and the levels of tumor necrosis factor-α(TNF-α) and interferon-γ(IFN-γ). The difference of kruüppel-like factor 4(KLF4) expression between SW480 and HCT116 cells was detected by Western blots, qPCR and ELISA. After pretreatment of HCT116 cells with KFL4 inhibitor Kenpaullone(Ken), the CMHCT116 and CMHCT116+Ken were co-cultured with M0 macrophages and the polarization of M2 macrophages was observed by flow cytometry, qPCR and ELISA. The CMHCT116-Mφ and CM_((HCT116+Ken)-Mφ) was co-cultured with HCT116 cells, and the PD-L1 expression of HCT116 cells was detected by Western blot and qPCR. Results After the stimulation of M0 macrophages with CM, the proportion of CD11b+CD206+ cells in HCT116-Mφ cells was higher, and the expression of M2 macrophage markers interleukin(IL-10) and transforming growth factor-β(TGF-β) were higher. Compared with the CMSW480-Mφ group, the PD-L1 protein expression level was higher in the CMHCT116-Mφ group. After co-culture with T lymphocytes, the cell survival rate are the most in CMHCT116-Mφ group, while the levels of TNF-α and IFN-γ were the lowest. After the addition of Ken, the polarization ratio and markers of M2 macrophages decreased. Compared with CMHCT116-Mφ group, the expression of PD-L1 in HCT116 cells of the CM_((HCT116+Ken)-Mφ) group decreased. Conclusion Highly metastatic colorectal cancer cells induce polarization of M2 macrophages by secreting KLF4, promote PD-L1 expression in colorectal cancer cells, facilitate tumor immune escape, and provide potential targets for clinical immunotherapy.

abstract:
Objective To investigate the expression and biological function of circular RNA(circRNA) hsa＿circ＿0002938 in gastric cancer. Methods 45 pairs of tumor tissues and para-cancerous tissues from gastric cancer patients who underwent tumor reduction surgery at the hospital were collected. Human gastric cancer cells SGC-7901, BGC-823, HGC-27, MGC-803 and immortalized gastric mucosal epithelial cells GES-1 were routinely cultured. RT-qPCR was utilized to detect the expression of hsa＿circ＿0002938 in gastric cancer tissues and cells. Chi-square test was used to analyze its association with the clinicopathological characteristics of patients, and the Kaplan-Meier method was employed to assess the relationship between hsa＿circ＿0002938 expression levels and patient prognosis. The effects of hsa＿circ＿0002938 on the proliferation, migration and invasion abilities of gastric cancer cells were detected by CCK-8, scratch wound healing and Transwell assays. Bioinformatics was used to predict the miRNA and its downstream target genes that hsa＿circ＿0002938 might bind to. The competitive endogenous RNA regulatory network was constructed and the functions of the target genes were enriched. Results The expression of hsa＿circ＿0002938 in gastric cancer tissues and cells was much higher than that in para-cancerous tissues and GES-1 cells(P<0.05). By contrast, patients in the high hsa＿circ＿0002938 expression group had significantly shorter 2-year progression-free survival than that in the low expression group after surgery(P<0.01). Moreover, knocking down the expression of hsa＿circ＿0002938 reduced the proliferative, migratory, and invasive abilities of gastric cancer cells(P<0.05). Bioinformatics predictive analysis showed that hsa＿circ＿0002938 could bind to hsa-miR-342-3p and hsa-miR-503-5p. The downstream target genes of miRNA were involved in several cancer-related pathways, like mitogen-activated protein kinase(MAPK), hippo and Wnt signaling pathway. Conclusion Hsa＿circ＿0002938, which is highly expressed in gastric cancer tissues and cells, is closely associated with poor patient prognosis. Knockdown of hsa＿circ＿0002938 inhibits the proliferation, migration, and invasion abilities of gastric cancer cells. The study reveals the potential role of hsa＿circ＿0002938 in the progression of gastric cancer, offering new insights into its prevention and treatment.

abstract:
Objective To construct microRNA(miR)-22 gene knockout(miR-22-/-) mice using CRISPR/Cas 9 technology, to breed miR-22-/- mice and to identify their genotypes. Methods In this experiment, CRISPR/Cas 9 technology was used to construct miR-22-/- genetically engineered mice. After gene identification, the F0 generation miR-22-/- mice were mated with wild-type mice in the same litter to obtain F1 generation miR-22-/- mice. The miR-22 knockout efficiency was analyzed at the RNA level by real-time fluorescence quantitative polymerase chain reaction(qPCR). Western blot was used to detect the interaction between miR-22 and target genes. Results miR-22-/- mice were successfully constructed using CRISPR/Cas 9 technology, gene identification was performed on the bred mice, and three stable genotypes of miR-22+/+,miR-22+/-, and miR-22-/- were identified. The real-time fluorescence quantitative PCR detection results confirmed that miR-22-/- mice showed almost no expression of miR-22 in the heart, liver, lung, kidney, spleen, and thymus tissues compared to wild-type mice in the same litter. Western blot analysis showed that the relative expression level of NLRP3 protein in miR-22-/- mouse tissues was lower than that in wild-type mice. Conclusion A miR-22-/- mouse model is successfully constructed, and stable genetic homozygous miR-22-/- mice is obtained. This indicates that miR-22 has an inhibitory effect on the downstream target gene NLRP3.

abstract:
Objective To investigate the role and mechanism of long non-coding RNA(lncRNA) H19 in osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs) and bone regeneration in mice with foot fracture.Methods BMSCs were divided into microrNA-149-5 p(miR-149-5 p)-mimic negative control(miR-mimicNC) group and miR-149-5p-mimic(miR-mimic) group and transfected with IncRNA H19-wt and IncRNA H19,respectively.Mut recombinant plasmid,or bone morphogenetic protein 2(BMP2) 3'UTR-wt and BMP2 3' UTRmut recombinant plasmid were transfected,respectively.Dual-luciferase reporter gene assay was used to detect the luciferase activity of each group.To study the regulatory effect and mechanism of IncRNA H19 on osteogenic differentiation in vitro,BMSCs were divided into control group and osteogenic induction group(osteogenic group).After osteogenic induction,BMSCs were transfected with corresponding plasmids and divided into osteogenic+si-NC group,osteogenic+si-H19 group,osteogenic+si-H19+miR-inhibitor-NC group,osteogenic+si-H19+miR-inhibitor group,osteogenic+si-H19+pcDNA-NC group,and osteogenic group si-H19+pcDNA-BMP2 group.Osteogenic differentiation was evaluated by alkaline phosphatase(ALP) activity assay and alizarin red staining.A mouse foot fracture model was established,and 36 mice were randomly divided into sham operation group,model group,model+pcD-null group,and model+pcD-H19 group,with 9 mice in each group.Osteogenic differentiation was assessed by ALP activity assay.Real-time fluorescent quantitative PCR(qPCR) was used to detect the expression of IncRNA H19,miR-149-5p,and BMP2.Western blot was used to detect the expression of BMP2,OCN,OSX,RUNX2,and OPN.Results In cells transfected with IncRNA H19-wt,the luciferase activity in miR-mimic group was lower than that in miR-mimic-NC group(P

abstract:
Objective To investigate the expression and interaction of activating transcription factor 3(ATF3) and smad family member 4(Smad4) in pterygium. Methods In this study, two sets of data GSE51995 and GSE2513 in NCBI database were analyzed by bioinformatics methods to screen out the key difference-expressed gene ATF3 in pterygium when normal conjunctiva was used as the control. The differential genes identified by bioinformatics analysis were further detected by reverse transcription quantitative polymerase chain reaction(RT-qPCR). Finally, the literatures were searched and The Gene Transcription Regulation Database(GTRD) was used to predict whether there was a target gene Smad4 that might bind to the target gene. The expression of Smad4 in normal conjunctival and pterygium tissues was verified by RT-qPCR, and the interaction between the target protein ATF3 and Smad4 in the pterygium was investigated through Co-Immunoprecipitation(Co-IP). Results The results of bioinformatics analysis showed that compared with normal conjunctival tissue, there was significantly different expression of ATF3 gene in pterygium tissue(P

abstract:
Objective To investigate the correlation between the levels of calcitonin gene-related peptide α(CGRPα) and the poor responses to the treatment of phenylephrine in pregnant women with hypotension during caesarean section under intraspinal anesthesia.Methods A total of 87 pregnant women who underwent intraspinal anesthesia cesarean section were recruited.They were grouped according to the presence or absence of hypotension after anesthesia and the response to ephedrine and Phe.As to Anesthesia-0 group,pregnant women had no hypotension,35 cases(40.2%);Anesthesia-1 group,pregnant women had hypotension and good response to ephedrine treatment,12 cases(13.8%);Anesthesia-2 group,pregnant women had hypotension and had poor response to ephedrine treatment,6 cases(6.9%);Anesthesia-3 group,pregnant women had hypotension and good response to Phe treatment,8 cases(9.2%);Anesthesia-4 group,pregnant women had hypotension and no response to Phe treatment,26 cases(29.9%).The levels of CGRPα in serum and placental tissues of pregnant women were determined by ELISA.The rats pregnant normally.A hypotension rat model was established by intragastric administration of risserpine.50 rats were randomly divided into control group,model group,pregnancy group,model+pregnancy group,model+pregnancy+phenylephrine group,with 10 rats in each group.Body weight of the rats was continuously monitored during the 2nd to 6th week of risserpine administration.Systolic blood pressure was measured at the end of the 6th induction week.And the peripheral blood sample and placental tissues were collected and the levels of CGRPα were determined.Results The detection of CGRPα levels in the pregnant women serum and placental tissues showed that compared with Anesthesia-0 group,CGRPα levels in Anesthesia-1-4 groups serum were higher(P 0.05).At the end of the 6th week,compared with pregnancy group,systolic blood pressure decreased in model+pregnancy group and model+pregnancy+phenylephrine group(P

abstract:
Objective To explore the relationship between the replication-associated bromodomain protein 4(BRD4)and human papillomavirus(HPV) 16(HPV16) viral load in cervical squamous cell carcinoma and CINⅠtissues, confirm the effects of BRD4 degradation agent MZ1 on viral load. Methods Thirty HPV16-positive cervical cancer specimens and 30 non-cervical cancer specimens were collected, and the viral load of the samples was detected by real-time fluorescence quantitative PCR, and the expression of BRD4 was analyzed by immunohistochemistry and Western blot. Results The viral load was higher in the samples of cervical cancer group than in the samples of non-cancer group, the difference is statistically significant(P

abstract:
Objective To investigate the role of endoplasmic reticulum stress in the occurrence and development of fatty liver induced by high fat. Methods In the high-fat Drosophila model, the high-fat group was fed with high-fat medium, while the control group was fed with normal medium; in the mouse fatty liver model, the high-fat group was fed with high-fat diet, and the control group was fed with normal diet; in the HepG2 cell steatosis model, the high-fat group was induced by palmitic acid(PA), and the control group was cultured with DMEM. The fat body size of the third instar larvae of Drosophila melanogaster was photographed. Steatosis in mice liver and HepG2 cells was observed by H&E and Oil Red staining. The expression levels of ATF6, Bip and CHOP in the third instar larvae, liver tissues of mice and HepG2 cells were analyzed by quantitative real-time polymerase chain reaction(qPCR) and Western blot. Results In Drosophila model, fat body and fat storage were obviously increased in high fat fed flies when compared with control group. The formation of liver fat droplets and cells vacuolation were confirmed by H&E and Oil Red staining in mice livers fed with high fat and HepG2 cells with palmitic acid treatment. The expression levels of ATF6, Bip and CHOP were significantly increased in third instar larvae and mice livers fed with high fat and palmitic acid treated HepG2 cells with palmitic acid treatment. Conclusion High fat may induce the occurrence and development of hepatic steatosis by activating endoplasmic reticulum stress.

abstract:
Objective To explore the relationship between the nodes of anxiety and depression symptoms in pregnant women,and to analyze the interaction between these symptoms.Methods A convenient sampling method was used to evaluate the level of anxiety and depression among pregnant women who underwent antenatal check-up in the obstetric examination in a Grade Ⅲ A hospital.Data were collected using the General Data Questionnaire,the Generalized Anxiety Disorder Scale(GAD-7),and the Patient Health Questionnaire Depression Scale(PHQ-9).Based on the relationship between anxiety and depressive symptoms in pregnant women,a network was constructed and analyzed.Results Among the 1 742 subjects,476(27.3%) had anxiety,290(16.6%) had depression,and 218(12.5%) had anxiety and depression comorbidities.The univariate analysis revealed statistically significant differences in various factors such as age,education level,occupation,gestational week,sleep status during pregnancy,exercise status during pregnancy,drinking,knowledge of pregnancy,planned pregnancy,living environment,husband-wife relationship,parent-in-law relationship and parental relationship had statistically significant differences in whether they had CAD(all P

abstract:
Objective To explore the causal relationship between 46 phenotypes(including 15 seropositive case-control phenotypes and 31 quantitative antibody-measurement phenotypes) and ulcerative colitis(UC) using two-sample bidirectional Mendelian randomization(TSMR). Methods Single nucleotide polymorphisms(SNPs) significantly associated with the relative abundance of the 46 antibody sera were extracted as instrumental variables according to preset thresholds. Summary statistics for UC were obtained from the OPEN GWAS database(n=47 745). MR-Egger regression, weighted median method(WME), inverse variance weighting(IVW), the simple mode method(SM), and weighted multitude method(WM) were used to estimate the causal relationship between antibody levels and UC, primarily using the IVW method. The results were assessed according to the effect indicator dominance ratios(OR) and 95% confidence intervals(CI). Sensitivity analysis, heterogeneity test, gene pleiotropy test, and outlier test(MR-PRESSO) were combined to verify the stability and reliability of the results, and the causal association study was performed again using reverse Mendelian randomization(MR). Results IVW results showed that Epstein-Barr(EB) virus EA-D antibody levels(OR=0.806, 95%CI=0.693-0.939, P

abstract:
Objective This study employs metabolomics to analyze the characteristic biomarkers and metabolic pathways in the serum of pregnant women with positive thyroid peroxidase antibodies(TPOAb) during early pregnancy. The objective is to explore the relationship between thyroid dysfunction and maternal-fetal health. Methods Early-pregnancy women undergoing antenatal check-ups for thyroid function and antibody testing at our hospital were selected. According to the TPOAb results, participants were categorized into a TPOAb-positive group and a TPOAb-negative group. The serum metabolic profiles were analyzed using liquid chromatography-tandem mass spectrometry(LC-MS/MS) technology to identify differences between the two groups. The Kyoto Encyclopedia of Genes and Genomes(KEGG) database was utilized for metabolic pathway enrichment analysis of differential metabolites. Results A total of 79 significantly different metabolites were identified in the serum of TPOAb-positive pregnant women compared to the control group, including 20 upregulated and 59 downregulated metabolites. KEGG enrichment analysis indicated that these differential metabolites were mainly involved in 21 key metabolic pathways. Among the metabolites associated with TPOAb, 31 were identified, with 6 showing positive correlation and 25 showing negative correlation. Metabolic pathway enrichment analysis revealed that these differential metabolites were closely related to Glycerophospholipid metabolism, Valine, leucine and isoleucine biosynthesis, Glycosylphosphatidylinositol(GPI)-anchor biosynthesis, and Glycerolipid metabolism pathways. Conclusion Significant differences in metabolites and their associated metabolic pathways are identified in the serum of TPOAb-positive pregnant women during early gestation, indicating that these metabolite alterations are closely linked to thyroid dysfunction and maternal-fetal health.

abstract:
Objective To assess and to compare the diagnostic performance of cervical ultrasound and 18F-fluorodeoxyglucose(18F-FDG PET/CT) in identifying cervical lymph node metastases in patients with differentiated thyroid cancer(DTC) following total thyroidectomy, focusing on both early and delayed imaging phases. Methods A retrospective review was performed on 83 DTC patients with DTC who had undergone total thyroidectomy. A total of 143 cervical lymph nodes, comprising both metastatic and non-metastatic nodes, were evaluated using 18F-FDG PET/CT and ultrasound. Sensitivity, specificity, and overall diagnostic accuracy were calculated for each imaging modality. Differences in lymph node detection rates across cervical regions using ultrasound were analyzed, along with nodal size parameters(long and short axis diameters), to investigate potential limitations of ultrasound in this context. Additionally, PET/CT parameters including axial ratio, early and delayed maximum standardized uptake values(SUVmax), and changes in SUVmax between imaging timepoints(ΔSUVmax) were compared between metastatic and non-metastatic lymph nodes to evaluate the diagnostic value and underlying mechanisms of PET/CT in this clinical setting. Results 18F-FDG PET/CT demonstrated a sensitivity of 91.58%(98/107), a specificity of 72.22%(26/36), and an overall diagnostic accuracy of 86.71%(124/143) in identifying cervical lymph node metastases post-total thyroidectomy in DTC patients. In comparison, ultrasound showed a lower sensitivity of 50.47%(54/107), but a higher specificity of 97.22%(35/36), with an overall accuracy of 62.24%(89/143). Notably, ultrasound exhibited a significantly higher false-negative rate in the first cervical lymph node group compared to the second group [56.6%(30/53) vs 43.4%(23/53); χ2= 20.396, Pmax compared to early-phase imaging(Z=-8.147, Pmax threshold of 2.5, delayed imaging identified a greater number of positive lymph nodes than early imaging(χ2=18.127, Pmax values were significantly higher in malignant nodes compared to benign ones(Z=-3.689, Pmax was markedly elevated in metastatic lymph nodes(Z=-4.189, P18F-FDG PET/CT proves to be superior to ultrasound in diagnosing cervical lymph node metastasis in patients with differentiated thyroid cancer(DTC) following total thyroidectomy. Delayed-phase imaging offers higher tumor-to-background ratio, which enhances the visibility of metastatic lymph nodes. In addition, dual-phase 18F-FDG PET/CT plays greater advantages in the differential diagnosis between metastatic lymph nodes and non-metastatic lymph nodes.

abstract:
Objective To establish an enzyme-linked immunosorbent assay(ELISA) for exosome B7-H3 in plasma, and to explore the clinical significance and diagnostic value of exosome B7-H3 in plasma from non-small cell lung cancer(NSCLC). Methods The plasma of 70 NSCLC patients(NSCLC group) and 36 healthy controls(HC group) were collected. Exosomes and microvesicles in plasma were separated by ultra-fast centrifuge method, and the expression levels of B7-H3 in plasma exosomes in NSCLC groups and HC groups were compared by Western blot method. In NSCLC group, the expression levels of B7-H3 in plasma exosomes and microvesicles in NSCLC group were compared. A simple and feasible ELISA method was established to detect the expression level of exosome B7-H3 in plasma by means of polyethylene glycol(PEG) precipitation and its clinical significance was analyzed. Logistic regression model was established to predict plasma-derived exosome B7-H3 as a risk factor, and receiver operating characteristic curve(ROC) was used to investigate the diagnostic value of exosome B7-H3 in NSCLC. Results For exosomes and microvesicles in plasma which were extracted by ultracentrifugation, Western blot results showed that the expression level of B7-H3 in plasma exosomes of NSCLC group was higher than that of HC group(P=0.032), and the expression level of B7-H3 in plasma exosomes was higher than that of microvesicles of NSCLC group(P=0.012). The expression level of exosome B7-H3 in plasma extracted by PEG precipitation was also higher in NSCLC group than that in HC group(P=0.024). The expression level of exosome B7-H3 in plasma of NSCLC patients was not related to gender, age, smoking or pathological type, but was related to T stage(P=0.002), N stage(P

abstract:
Objective To explore the utility of a nomogram integrating contrast-enhanced CT radiomics with clinical features in the preoperative prediction of WHO/ISUP grade for clear cell renal cell carcinoma(ccRCC). Methods A total of 214 patients with pathologically proven ccRCC who underwent enhanced CT scan before surgery were retrospectively included. According to the WHO/ISUP grade system, the cases were classified into low-grade(grades Ⅰ-Ⅱ) and high-grade(grades Ⅲ-Ⅳ), and then randomly divided into training and test set with a ratio of 4 ∶1. Regions of interest were segmented from both unenhanced and three-phase enhanced images, and radiomic features were extracted. Feature selection and dimensionality reduction were performed using Spearman rank correlation coefficients and LASSO regression, followed by the construction of the radiomic model with the KNN algorithm. Clinical and semantic imaging features were selected through univariate and multivariate analyses, and a clinical model was developed using the KNN algorithm. The clinical and radiomics signatures were used to construct a combined model and a nomogram was developed. The ROC curve and delong test were used to evaluate the diagnostic performance of the model, while calibration and decision curve analyses assessed its accuracy and clinical applicability. Results 8 clinical features and 11 radiomic features were selected. The combined model, integrating these clinical and radiomics signatures, exhibited robust predictive performance with AUC values of 0.887 in the training set and 0.800 in the test set. The calibration curve demonstrated good consistency between the nomogram model and actual outcomes, while decision curve analysis indicated a favorable net benefit for the nomogram. Conclusion The nomogram constructed by combining radiomics and clinical signatures can provide evidence for preoperative prediction of ccRCC grade and guide clinical decision-making.

abstract:
Objective To compare the antiviral efficacy and renal safety of nucleoside analogs(NAs) monotherapy versus combination therapy in hepatitis B e antigen(HBeAg)-positive chronic hepatitis B(CHB) patients with high viral load. Methods This study enrolled a total of 353 treatment-na6ve HBeAg-positive chronic hepatitis B(CHB) patients with high viral load, the treatment regimen was divided into 5 groups, consisting of 4 monotherapy groups and 1 combination therapy group as follows: 88 cases in the Entecavir(ETV) group, 135 cases in the Tenofovir Disoproxil Fumarate(TDF) group, 34 cases in the Tenofovir Alafenamide Fumarate(TAF) group, 25 cases in the Tenofovir Amibufenamide(TMF) group, and 71 cases in the ETV combined with TDF(ETV+TDF) group. A retrospective cohort study design was adopted to analyze HBV DNA levels, serological indicators(HBsAg and HBeAg levels), renal function indicators(serum Scr levels, eGFR) at 24 and 48 weeks of treatment across various groups, as well as the HBsAg clearance rates, HBeAg seroconversion rates and HBV DNA suppression rates(HBV DNA0.05). At 48 weeks, the HBV DNA levels in the ETV+TDF combination therapy group was significantly lower than those in all monotherapy groups(P0.05). The HBV DNA level in the ETV group was higher than those in the remaining four groups(P0.05). Conclusion For HBeAg-positive chronic hepatitis B(CHB) patients with high viral load, the combination therapy of ETV and TDF significantly enhances viral suppression compared to monotherapy, without increasing the risk of renal adverse events. This suggests that the combination therapy can be considered a preferred strategy for this specific patient population.

abstract:
Objective To analyze the differential expression profile of miRNAs in amniotic fluid exosomes of fetuses with Down syndrome(DS) and provide insights for identifying novel biomarkers for the prenatal diagnosis of DS. Methods Amniotic fluid samples were collected from fetuses with DS and chromosomally normal fetuses. Exosomes were isolated from the amniotic fluid and subjected to high-throughput sequencing. Differentially expressed miRNAs were identified, and target genes were predicted using TargetScan and miRanda. Target genes located on chromosome 21 were selected, and their biological functions and associated diseases were analyzed using GeneCards, HGNC, NCBI Gene, UniProtKB/Swiss-Prot, Ensembl, and OMIM databases. GO and KEGG enrichment analyses were performed to investigate the biological functions of the enriched genes. Results A total of 59 differentially expressed miRNAs were identified, including 31 upregulated and 28 downregulated miRNAs. Based on a fold change>2 andP<0.05, 10 upregulated and 9 downregulated miRNAs with the highest expression levels were selected. Key miRNAs included hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7b-3p＿1ss22CT, and hsa-miR-199b-5p, withBACH1andIFNAR1identified as their shared target genes. GO analysis revealed that the enriched target genes were primarily involved in protein binding, metal ion binding, transferase activity, DNA binding, transcriptional regulation by RNA polymerase Ⅱ, and nucleotide binding. KEGG pathway analysis indicated that the target genes were mainly associated with metabolic pathways, cancer-related pathways, the PI3K-Akt signaling pathway, and the Rap1 signaling pathway. Conclusion Differential expression of miRNAs in amniotic fluid exosomes was observed between DS fetuses and those with normal karyotypes. Combined analysis with placental miRNAs revealed hsa-miR-199b-5p as a common differentially expressed miRNA in both DS amniotic fluid and placenta. It is hypothesized thatBACH1andIFNAR1, shared target genes of hsa-miR-199b-5p, hsa-let-7b-5p, hsa-let-7c-5p, and hsa-let-7b-3p＿1ss22CT, may play a role in the pathogenesis of DS.

abstract:
Polycystic ovarian syndrome(PCOS) is a very common endocrine and reproductive disease. Its etiology and pathogenesis are complex and not yet fully clear. At present, the clinical treatment is mainly symptomatic. Studies have revealed that ferroptosis, as a new form of cell death, may play a key regulatory role in the occurrence and development of PCOS. In addition, there is an increase in autophagy/mitochondrial autophagy in PCOS patients, which may be closely related to the occurrence of ferroptosis. This review summarizes the pathogenesis of mitochondrial autophagy and ferroptosis in PCOS, and analyzes the interrelationship between mitochondrial autophagy and ferroptosis in granulosa cells, in order to provide new insights and potential therapeutic targets for the clinical treatment of PCOS.

abstract:
Breastfeeding has an irreplaceable effect on the growth and development of premature infants and the reduction of complications during hospitalization. In recent years, with the opening of China′s birth policy, the incidence of premature babies has increased, and the corresponding complications have also gradually increased. Breast milk not only carries a rich supply of nutrients, including rich proteins, lipids, sugars to support the normal growth and development of infants, but also contains a large number of immunoglobulins, hormones and other bioactive substances to promote the improvement of body immune function and the development of nervous system. In recent years, many studies have been carried out on the current situation of breast milk operation in breast milk bank, the collection and influencing factors of breast milk, the strategies of breast milk enrichment and its effects on the growth and development of premature infants and complications. Therefore, breast-feeding and intensive breast-feeding can effectively promote the growth and development of premature infants, and reduce the occurrence of complications.

abstract:
Skin, the largest organ of the human body, serves as the primary barrier against environmental hazards. Skin aging is a complex process influenced by both intrinsic factors, such as endogenous physiological changes, and extrinsic factors, including environmental exposures, reflecting the internal health status of the body. During skin aging, various cell types exhibit diverse responses to aging-related stimulations. This paper systematically reviews the roles of cells in the pathogenesis of skin aging and explores potential therapeutic strategies for mitigating this process. Furthermore, based on the biological activities of cells during skin aging, we elucidate the interaction mechanisms among cells within the skin microenvironment.