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Objective To investigate the role of Toll-like receptor 4(TLR4) in hepatocyte regeneration after acetaminophen(APAP)-induced injury in human normal liver cell(L02) and its possible mechanism. Methods L02 cells were culturedin vitro, and cell viability was detected by CCK-8 assay. The optimal concentration and duration of APAP and the concentration of TLR4 inhibitor(TAK-242) were determined. The protein expression levels of nuclear factor-κB(NF-κB), microtubule-associated protein light chain 3(LC3), p62, receptor interacting protein kinase 1(RIP1), receptor interacting protein kinase 3(RIP3), signal transducer and activator of transcription 3(STAT3), phosphorylation of STAT3(p-STAT3), proliferating cell nuclear antigen(PCNA) and Cyclin D1 were detected by Western blot. The mRNA expression levels of TLR4, NF-κB, tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), interleukin-1β(IL-1β), PCNA, Cyclin D1 and Ki67 were detected by qRT-PCR. Results According to the results of CCK-8, L02 cells were treated with 5 mmol/L APAP for 24, 36, 48 h to simulate liver injury and regeneration modelin vitro, and TAK-242 100 nmol/L was pretreated 2 h before APAP to inhibit TLR4. Compared with the control group, the protein levels of NF-κB, RIP1, p-STAT3, PCNA, Cyclin D1 and the mRNA levels of TNF-α, IL-1β and PCNA increased in the APAP 24 h group; the protein levels of NF-κB, RIP1, RIP3, p-STAT3, PCNA, Cyclin D1 and the mRNA levels of TLR4, NF-κB, TNF-α, IL-1β, PCNA and Cyclin D1 increased in the APAP 36 h group; the protein levels of NF-κB, RIP1, p-STAT3, PCNA, Cyclin D1 and the mRNA levels of TLR4, NF-κB, TNF-α, IL-1β, IL-6, PCNA, Cyclin D1 and Ki67 increased in the APAP 48 h group. The protein levels of NF-κB, RIP1, RIP3, p-STAT3, PCNA, Cyclin D1 and the mRNA levels of TLR4, NF-κB, TNF-α, IL-1β, IL-6, PCNA, Cyclin D1, Ki67 significantly decreased in APAP+TAK-242 24 h and 48 h group than the APAP group at the same time point; the protein levels of NF-κB, PCNA and the mRNA levels of TLR4, NF-κB, TNF-α, IL-1β, IL-6, PCNA and Ki67 in APAP+TAK-242 36 h group were also significantly lower than those in APAP 36 h group. Compared with the control group, autophagy was activated in the APAP group, while autophagy was inhibited in the APAP+TAK-242 group. Conclusion TLR4 may affect the TLR4/NF-κB pathway, up-regulate the levels of inflammatory factors and autophagy, and promote hepatocyte regeneration after APAP-induced liver injury in L02 cells.

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Objective To explore the effects of exposure to lipopolysaccharides in late pregnancy on age-related cognitive changes in offspring of mice, and to investigate whether there is a gender specific genetic effect. Methods Institute of cancer research(ICR)CD-1 mice during gestational days 15-17 were injected with lipopolysaccharide daily(LPS group, 50 μg/kg), or equal volume of normal saline(CON group). At the age of 2 months after their delivery, LPS treated offspring mice(F1-LPS, male and female) were randomly selected and hybridized with age-matched wild-type CD-1 mice. F1-LPS males and females with different littermates, and F1-CON males and females were hybridized to obtain F2 generations of different lineages. Similarly, F2-LPS mice were mated with wild-type mice to conceive the F3 generation. At the age of 3 and 18 months old, F1, F2, and F3 mice(n=8 in each group) were randomly selected to complete the Morris maze experiment in order to test their cognitive abilities. Results Compared with 3-month-old CON mice, 18-month-old CON mice showed poorer learning and memory abilities, especially in females. For F1 generation, the learning and memory abilities of the 3-month-old and 18-month-old F1-LPS mice were inferior to those of the same aged CON mice. For F2 generation, the 3-month-old F2-LPS-parental mice had poorer learning and memory compared to the same aged CON mice, while the F2-LPS-paternal mice only had poorer memory compared to the same aged CON group. The learning and memory abilities of 18-month-old F2-LPS paternal and F2-LPS-parental mice were inferior to those of the same aged CON mice. The learning and memory abilities of F2-LPS maternal male mice were inferior to those of CON male mice, and the memory abilities of F2-LPS maternal mice were stronger than those of F2-LPS-parental mice. With regards to the F3 generation, the memory of the 3-month-old F3-LPS-parental mice was poorer than that of the same aged CON mice. The learning and memory abilities of F3-LPS paternal and F3-LPS-parental mice at 18 months old were inferior to those of CON mice of the same age. The 18-month-old F3-LPS maternal and paternal male mice had better memory than F3-LPS-parental male mice. Conclusion Exposure to lipopolysaccharides in late pregnancy can accelerate age-related cognitive decline in offspring mice, and it has a cross generational genetic effect and gender differences, mainly in paternal inheritance.

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Objective To investigate the effects and mechanisms of oridonin(ORI) on human keloid-derived fibroblasts(HKF). Methods The effects of ORI on the proliferation activity of HKF were assessed using the CCK-8 assay. The experiment was divided into control and experimental groups. Cell scratch and Transwell assays were conducted to evaluate the migration and invasion capabilities of HKF. Real-time quantitative PCR(RT-qPCR) and Western blot were used to examine the impact of ORI on the expression of extracellular matrix-related mRNA and fibronectin 1(FN1), α-smooth muscle actin(α-SMA), type I collagen( COL I), and COL Ⅲ in HKF. The experiment was also divided into control, model, and transforming growth factor(TGF)-β1+ORI groups. RT-qPCR and Western blot were utilized to determine the effects of ORI on the expression of TGF-β1-induced mRNA and nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3), apoptosis-associated speck-like protein containing a CARD(ASC), Smad2, Smad3, phosphorylated Smad2(p-Smad2), and p-Smad3 in HKF. Results CCK-8 assay demonstrated that the cell inhibition rate of HKF progressively increased with increasing concentrations of ORI. Compared with the control group, the experimental group exhibited a significant reduction in the migration area at 24 hours and a decrease in the number of invasive cells. Furthermore, there was a significant downregulation in the expression levels of FN1, α-SMA, COL I, and COL Ⅲ(P<0.05). In comparison with the control group, the model group showed a significant upregulation in the expression of NLRP3, ASC, Smad2, Smad3, p-Smad2, and p-Smad3(P<0.05). Relative to the model group, the TGF-β1+ORI group demonstrated a significant downregulation in the expression of NLRP3, ASC, Smad2, Smad3, p-Smad2, and p-Smad3(P<0.05). Conclusion ORI the proliferation, migration, and invasiveness of HKF, as well as the formation and deposition of the extracellular matrix, through the blockade of the TGF-β1/Smad signaling pathway and the NLRP3-mediated inflammatory response.

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Objective To investigate the expression of miR-142-5p in oral squamous cell carcinoma(OSCC) tissues and cell lines and its effects on oral squamous cell proliferation, migration, invasion and angiogenesis. Methods Sixteen groups of oral tumour tissues and paraneoplastic tissues were collected, and qRT-PCR was applied to detect the expression of miR-142-5p in the tissues. The effects of miR-142-5p on cell proliferation, migration, andinvasion were observed by cell counting kit-8(CCK-8), cloning, wound healing, Transwell, invasion assays, and the effect of miR-142-5p on angiogenesis was also detected by lumen formation assay.The expression of angiogenesisrelated proteins vascular endothelial growth factor(VEGFA), vascular endothelial calreticulin(VE-cadherin), epithelial calreticulin(E-cadherin), matrix metalloproteinase 2(MMP2), and matrix metalloproteinase 9(MMP9) was detected by Western blot after overexpression of miR-142-5p. Results miR-142-5p was lowly expressed in oral tumour tissues and cell lines. CCK-8 and clonogenic assays showed that miR-142-5p was inversely correlated with the proliferation of OSCC cells, wound healing and Transwell assays showed that miR-142-5p was inversely correlated with the migration of OSCC cells, and cell invasion assays showed that miR-142-5p was conversely correlated with the invasion of OSCC cells. Analysis of lumen formation assay showed that overexpression of miR-142-5p reduced the tube length and nodes of HUVECs. Western blot assay showed that up-regulation of miR-142-5p inhibited the VEGFA, VE-cadherin, MMP2, MMP9 expression and promoted E-cadherin expression. Conclusion Overexpression of miR-142-5p inhibites the proliferative, migratory and invasive effects of oral squamous carcinoma cells as well as angiogenesis, suggesting that miR-142-5p is a novel target for anti-tumour angiogenesis and against oral squamous carcinoma.

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Objective To investigate the effects of zalcitabine(ddC), a mitochondrial DNA polymerase γ(POLG) inhibitor, on the migration, invasion, and to preliminarily explore mitochondrial biogenesis of human triple-negative breast cancer MDA-MB-231 cells. Methods The effect of ddC on cell viability was detected using the MTT assay. The migration and invasion abilities of the cells were evaluated using the cell scratch and Transwell invasion assays. Cell apoptosis was determined using flow cytometry and a V-FITC/PI cell apoptosis detection kit. The protein expression of POLG, NADH dehydrogenase subunit Ⅰ(NADH1), NADH dehydrogenase subunit Ⅱ(NADH2), ATP synthase subunit 6(ATPase6), cytochrome c oxidase subunit Ⅰ(COX-1) and cytochrome c oxidase subunit Ⅲ(COX-3) were determined using Western blot. The POLG mRNA level and mtDNA copy number were determined using qPCR. The mitochondrial content and ATP levels were determined using MitoTracker Green fluorescent probe staining and an ATP determination kit. MDA-MB-231 cells were transfected with pcDNA3.1-EGFP-POLG plasmids to overexpress POLG. The inhibitory effects of ddC on cell migration and invasion were detected in POLG-overexpressed MDA-MB-231 cells. Results POLG expression was higher in MDA-MB-231 cells than in normal mammary epithelial cells(MCF-10A)(P<0.01). ddC inhibited cell viability in a dose-dependent manner. ddC inhibited the migration(P<0.01) and invasion(P<0.01) of MDA-MB-231 cells; however, it displayed no significant inhibitory effects on cell viability in normal mammary epithelial cells(MCF-10A) at the same concentration. ddC downregulated the protein(P<0.01) and mRNA(P<0.01) levels of POLG, reduced mtDNA copy number(P<0.01) and downregulated mtDNA-coded NADH1, NADH2, ATPase6, COX-1 and COX-3 protein expression(P<0.01) in MDA-MB-231 cells. Furthermore ddC inhibited mitochondrial content(P<0.01) and ATP(P<0.01) levels in MDA-MB-231 cells. POLG overexpression increased the migration(P<0.05) and invasion(P<0.05) abilities of MDA-MB-231 cells, while ddC did not significantly inhibit the migration and invasion abilities of MDA-MB-231 cells overexpressing POLG. Conclusion ddC downregulates POLG expression in MDA-MB-231 cells and inhibits mitochondrial biogenesis and ATP levels, thereby inhibiting the migration and invasion of MDA-MB-231 cells.

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Objective To investigate the effect of dexmedetomidine(Dex) on the release of glial cell line-derived neurotrophic factor(GDNF) during the anti-ischemic stroke. Methods SD rats were randomly divided into sham group(Sham group), sham + dexmedetomidine group(Sham+Dex group), ischemic stroke group(IS group), ischemic stroke + dexmedetomidine group(IS+Dex group), ischemic stroke + dexmedetomidine group +anti-GDNF(IS+Dex+anti-GDNF group), with 15 rats in each group. IS model was established by Longa′s methods. 24 hours later, the neurological scores were evaluated. Then, the rats were sacrificed and the cerebral spinal fluid, as well as the peripheral blood, were collected to detect the content of GDNF. In addition, TTC staining was used to evaluate the area of cerebral ischemic infarction, and immunofluorescence was applied to detect the activation of astrocytes. Furthermore, the activation phenotype of astrocytes was detected by qRT-PCR. Results Compared with Sham group, there were no significant changes in all indexes of rats in Sham+Dex group. In IS group, the neurological score increased(P<0.05), the local infarction area appeared, the release of GDNF in peripheral and central system increased(P<0.05), and the microglia activation was induced by IS, and the expression of neurotoxic and neuroprotective phenotype genes increased significantly(P<0.05). Compared with the IS group, the neurological function score of the rats in the IS + Dex group significantly decreased(P<0.05), the local infarct area was significantly reduced(P<0.05), and the release of GDNF further increased(P<0.05). The expression of neurotoxic astrocyte phenotype genes significantly decreased(P<0.05), while the expression of neuroprotective astrocyte phenotype genes further increased(P<0.05). However, intervention with anti-GDNF antibody obviously reversed the therapeutic effect of Dex compared with the IS + Dex group.Conclusion Dex can inhibit the phenotype of neurotoxic astrocytes, increase the proportion of neuroprotective astrocytes, further promote the release of GDNF, and play an anti-ischemic stroke role.

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Objective To investigate the mechanism of purinergic ion channel receptor 7(P2X7) intervention in reducing hippocampal damage in epileptic convulsions. Methods A total of 28 SD female rats were divided into 4 groups(n=7 in each group) according to random number table method. The male and female rats were combined with 2 ∶1 cage. The sperm cells observed by vaginal smear on day 2 were marked as day 0 of pregnancy(GD0), and the model of epileptic pregnant mice was prepared by injecting LPS into the tail vein of GD14 and PTZ into the abdomen continuously on GD16-18. The inhibitor group was prepared by intraperitoneal injection of P2X7 inhibitor(A-438079) immediately after intraperitoneal injection of PTZ in GD18. They were divided into:(1) normal pregnancy group(P group);(2) Epilepsy group(LPS+PTZ group);(3) P2X7 inhibitor group(LPS+PTZ+A-438079 group);(4) Eclampsia control group(LPS+PTZ+NS group). Racine scoring criteria were used to determine the grade of epileptic seizure in rats to evaluate the degree of epileptic seizure in each group and record the duration of each phase. The expression levels of P2X7 and NLRP3 in the hippocampus of pregnant mice were detected by Western blot. The expression of TNF-α, IL-1β, PIGF and sFlt-1 inflammatory factors in serum of pregnant mice was determined by ELISA. Results The tic behavioral scores of rats in LPS+PTZ group and LPS+PTZ+NS group were significantly higher than those in P group. Compared with LPS+PTZ+A-438079 group, LPS+PTZ+NS group and LPS+PTZ+NS group, the first stage of seizure latency, the fifth stage of seizure latency and the duration of 1-4 stages of seizure decreased. Both the rate of stage 5 seizures and the duration of stage 5 seizures were prolonged. Western blot results showed that compared with LPS+PTZ group and LPS+PTZ+NS group, the expressions of P2X7 and NLRP3 in hippocampus of pregnant rats in LPS+PTZ+A-438079 group increased. ELISA results showed that compared with P group, serum inflammatory factor markers tumor necrosis factor-α(TNF-α) and interleukin(IL)-1β were detected in LPS+PTZ and LPS+PTZ+NS groups. Soluble FMS-like tyrosine kinase 1(sFlt-1), a marker of preeclampsia; placental growth factor(PIGF) increased, while P2X7 receptor inhibitors were used to reduce all indicators. Conclusion Intervention of P2X7 receptor may reduce the systemic inflammatory response of epileptic pregnant mice, reduce neurohippocampal damage, increase the convulsive threshold, and inhibit the occurrence of epileptic convulsions.

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Objective To isolate aAcinetobacter baumannii(Ab) phage from underground sewage, study its properties, and to provide a theoretical basis for phage treatment of Ab infection. Methods Double-layer agar technique was used to isolate phages by using Ab GY-6 as the host strain. Biological characterization and therapeutic effect of the phage was tested. Genetic information of the phage was analyzed. Results Ab phage Abgy202162 was isolated. Transmission electron microscopy(TEM) analysis showed that the morphology of Abgy202162 exhibited an icosahedral structure. Biological characteristic analysis showed that the optimal multiplicity of infection was 1, the latent period was 5 min, and the burst size was approximately 520 PFU per cell. In addition, Abgy202162 remained stable at different concentrations of chloroform, pH, and temperatures. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) analysis showed that it contained 10 proteins with molecular weights ranging from 15 to 100 ku. The double-stranded(ds) DNA genome of Abgy202162 consisted of 40 889 bp and its G+C content was 38.85%. It contained 47 open reading frames(ORFs), of which 26 had specific functions, but no virulence related genes or antibiotic resistance genes were found. Phylogenetic analysis showed that Abgy202162 was a new phage in the Autographiviridae family, Beijerinkvirinae subfamily, and Friunavirus genus. Abgy202162 showed the ability to prevent Ab infection in the Galleria mellonellain vivomodel. Conclusion The phage Abgy202162 has strong environmental tolerance and high safety, indicating its potential as an antibiotic alternative used in the treatment of infections caused by Ab.

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Objective To investigate the expression and clinical significance of CD155 in oral squamous cell carcinoma(OSCC) tissues, as well as its impact on migration, proliferation and invasion. Methods Immunohistochemistry(IHC) was used to analyze the expression of CD155 in OSCC tissues and its correlation with clinicopathological features and prognosis. The transcription levels of CD155 were assessed by Western blot and RT-qPCR. Cell experiments were conducted to evaluate the effects of CD155 on OSCC cells following transfection with CD155 siRNA. Results CD155 was predominantly expressed on the cell membrane in OSCC tissues, with higher expression levels compared to normal tissues(χ2=50.750, P<0.000 1). High CD155 expression was associated with Ⅲ+IV disease stages(χ2=25.488, P=0.001), poor differentiation(χ2=6.299, P=0.012), T3+T4 depth of invasion(χ2=23.820, P=0.001) and lymph node metastasis(χ2=7.830, P=0.005) in OSCC patients; survival analysis revealed a correlation between high CD155 expression and shorter patient survival(P<0.05). COX regression analysis identified lymph node metastasis as an independent factor affecting the survival of OSCC patients(P<0.05). Both CD155 protein expression and mRNA transcription levels were significantly upregulated in OSCC cells(P<0.05). Transfection with siRNA-CD155 in OSCC cells resulted in significantly reduced proliferation, migration and invasion abilities compared to the control group(P<0.05).Conclusion CD155 is highly expressed in OSCC tissues and is associated with poor patient prognosis. Modulating CD155 expression can influence the biological functions of OSCC cells, leading to and inhibition of proliferation, migration, and invasion.

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Objective To explore the action mechanism of long non-coding RNA(lncRNAs) lncRNA KCTD13-DT in oral squamous cell carcinoma(OSCC) and its potential interaction with transcription factor c-Myc, providing a potential diagnostic and therapeutic target for patients with OSCC. Methods The expression of lncRNA KCTD13-DT in OSCC and paracancerous tissues was detected by qRT-PCR. The effects of c-Myc overexpression and knockdown on human tongue squamous carcinoma cells HN6 and CAL27 were detected by qRT-PCR. Fluorescence in situ hybridization(FISH) assessed the localization of lncRNA KCTD13-DT in cells. A dual luciferase reporter gene was used to analyze the role of c-Myc in target binding to the promoter region of lncRNA KCTD13-DT. Stable cell lines with knockdown or overexpression of lncRNA KCTD13-DT were constructed in human OSCC cell lines HN6 and CAL27 by lentiviral infection, and the knockdown and overexpression efficiencies of lncRNA KCTD13-DT were detected by qRT-PCR. Cell proliferation changes were detected by growth curve assay, CCK-8 assay, colony formation assay, and cell migration was detected by scratch assay and Transwell. Results lncRNA KCTD13-DT expression level was reduced in OSCC tissues and OSCC cells(HN6, CAL27), and Western blot verified that after knocking down and overexpression of c-Myc in HN6 and CAL27, the qRT-PCR experiments showed that c-Myc negatively regulated lncRNA KCTD13-DT, and overexpression of c-Myc significantly down-regulated lncRNA KCTD13-DT; knockdown of c-Myc significantly up-regulated lncRNA KCTD13-DT levels. Dual luciferase reporter gene showed that c-Myc could target lncRNA KCTD13-DT, and c-Myc could be involved in regulating and repressing the transcriptional activity of lncRNA KCTD13-DT. FISH showed that lncRNA KCTD13-DT mainly existed in the nucleus. Growth curve assay, CCK-8 assay, cell scratch assay, Transwell, and colony formation assay showed that knockdown of lncRNA KCTD13-DT promoted the growth and proliferation of OSCC cells, and overexpression of lncRNA KCTD13-DT significantly inhibited the proliferation and migration of OSCC cells. Conclusion lncRNA KCTD13-DT is negatively regulated by c-Myc.Knockdown of lncRNA KCTD13-DT promotes cell proliferation, while overexpression of it inhibits cell growth.

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Objective To explore the impact of bisphenol A(BPA) exposure on liver lipid metabolism in C57BL/6J mice and uncover the mechanisms at work. Methods Male C57BL/6J mice, aged eight weeks, were stratified into six cohorts: a control group on a standard diet(ND), a group on a standard diet with low-dose BPA(BPA-50 ND), a group on a standard diet with high-dose BPA(BPA-500 ND), a control groupon a high-fat diet(HFD), a group on a high-fat diet with low-dose BPA(BPA-50 HFD), and a group on a high-fat diet with high-dose BPA(BPA-500 HFD). Dosages for the low-and high-dose BPA groups were 50 and 500 μg/(kg·d), respectively, administeredviagavage over a duration of 12 weeks. Hepatic tissue underwent histological examination through hematoxylin and eosin(HE) staining. Furthermore, the expression levels of miR-122-5p and miR-143-3p in hepatic tissue, in addition to interleukin(IL)-6 and IL-10 in peripheral serum, were quantitatively measured employing quantitative reverse transcription-polymerase chain reaction(qRT-PCR) and enzyme-linked immunosorbent assay(ELISA), respectively. Results Histopathological analysisviaHE staining indicated intact hepatic lobule architecture in the ND group, whereas other groups displayed variable degrees of lipid droplet accumulation and damage to hepatic lobules. Notably, supplementation with BPA, particularly in conjunction with a high-fat diet, led to a progressive increase in IL-6 levels and a decrease in IL-10 levels in peripheral blood. In the context of a standarddiet, an augmentation in BPA concentration corresponded with a decline in the expression of miR-122-5p and miR-143-3p. Conversely, within the high-fat diet cohort, enhanced BPA concentrations were associated with increased expressions of these microRNAs. Pearson correlation analysis disclosed a significant positive correlation between the expression of miR-122-5p and miR-143-3p and the level of IL-10 in the standard diet group(P<0.01). In the high fat diet group, the expression level of miR-122-5p was positively correlated with the concentration of IL-6(P<0.05), and the expression level of miR-143-3p was negatively correlated with the concentration of IL-10(P<0.05). Conclusion BPA can induce the occurrence and progression of nonalcoholic fatty liver disease(NAFLD) by regulating the expression of miR-122-5p and miR-143-3p and regulating the levels of inflammatory factors IL-6 and IL-10.

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Objective To investigate the role of growth arrest-specific 6(Gas6)/Mer tyrosine kinase(MerTK) signaling pathway in ferroptosis in diabetes retinopathy(DR). Methods Human retinal pigment epithelial cells(ARPE-19) were divided into control group, HG group, HG+sh-Gas6 group, and HG+Gas6 group. Cells were exposed to 25 mmol/L D-glucose for simulating anin vitrohyperglycemic(HG) environment. The control group was exposed to a 20 mmol/L mannitol+5.5 mmol/L glucose environment. Rats were randomly divided into normal control group, DR group, and DR+Gas6 group, with 20 rats in each group. A DR model was established by intraperitoneal injection of STZ solution. Cell proliferation was evaluated using the cell count kit 8(CCK-8) assay. Lipid reactive oxygen species(ROS) levels were measured by flow cytometry, and levels of malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione peroxidase(GSH-Px) were measured by biochemical assays to evaluate iron death. The expression of Gas6 and MerTK proteins was analyzed by Western blot. Results Compared with HG group, the cell viability, SOD, GSH-Px levels in HG+Gas6 group increased significantly(P<0.05), while the levels of lipid-ROS and MDA decreased significantly(P<0.05). In HG+sh-Gas6 group, the cell viability, SOD and GSH-Px levels decreased significantly(P<0.05), while the levels of lipid-ROS and MDA increased significantly(P<0.05). In addition, the expression of GPX4 protein in HG+Gas6 group was significantly higher than that in HG group(P<0.05), and the expression of GPX4 protein in HG+sh-Gas6 group was significantly lower than that in HG group(P<0.05). Compared with the control group, the average thickness of retinal nerve fiber layer in DR group significantly decreased(P<0.05), while that in DR+Gas6 group increased significantly(P<0.05). In addition, the levels of MDA and iron in retinal pigment epithelium(RPE) tissues of DR+Gas6 group decreased significantly(P<0.05), while the levels of GSH and the expressions of Gas6, MerTK and GPX4 proteins increased significantly(P<0.05). Conclusion HG treatment accelerates the clearance of GPX4 by inhibiting the Gas6/MerTK signaling pathway, inducing ferroptosis and cell growth inhibition in ARPE-19 cells. In addition, up-regulating the expression of Gas6/MerTK signal in DR rat retina can alleviate ferroptosis and oxidative stress in RPE tissue, and help to restore the average retinal nerve fiber layer thickness.

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Objective To understand the bioinformatics of human ribosomal protein L22(RPL22) gene, prokaryotic expression and purification of human RPL22 recombinant protein, synthesis of human RPL22(labeled as agent M)in vitro, study the effects of agent M on proliferation, cycle and apoptosis of NSCLC A549 cells, and to analyze the effect of combination chemotherapy with M and cisplatin(DDP). Methods RPL22 bioinformatics was analyzed. RPL22 gene was cloned by PCR, the single chain oligonucleotide was designed and synthesized to obtain the target gene, and connect the pET-28α vector. The pET-28α-RPL22 recombinant plasmid was constructed, and the receptive cell DH5αof Escherichia coli was transformed. The expression of recombinant protein RPL22 was analyzed by SDS-PAGE and Western blot. The semi-inhibitory rate(IC50) of M and DDP in A549 cells and the effect of M on the proliferation of A549 cells were detected by CCK-8. The effect of agent M on apoptosis and cycle of A549 cells was detected by flow cytometry. qPCR and Western blot assays were used to detect M and DDP on phosphatidylinositol-3-kinase/protein kinase B(PI3K/AKT) and adenosine 5′-monophosphate activation of protein kinase/mammalian target of rapamycin(AMPK/mTOR) signaling pathway, respectively. Results RPL22 protein was an unstable and hydrophilic protein. The recombinant protein was dissolved and purified to obtain high concentration of recombinant protein. In A549 cells, the IC50of reagent M was 400 μg/L, and the IC50of DDP was 10 μmol/L, and the concentration of reagent M was inversely proportional to the cell activity. The inhibitory effect of reagent M(200 μg/L) combined with DDP(2.5 μmol/L) was similar to that of DDP(10 μmol/L) alone. Reagent M could induce G2cell cycle arrest and promote apoptosis, and might be involved in the regulation of PI3K/AKT and AMPK/mTOR pathways. Conclusion The recombinant RPL22 protein is cloned and expressed successfully.

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Objective To investigate the effect of miR-199a-5p on common bile duct ligation(BDL)-induced liver fibrosis in rats by regulating intestinal flora. Methods The 25 SD rats were randomly divided into five groups: the Sham group, the BDL group, the negative control adenovirus(NC adv) group, the miR-199a-5p adv group and the miR-199a-5p sponge adv group. The pathological changes of liver tissue and the degree of liver fibrosis were observed by HE, Masson and Sirius Red staining. The levels of aspartate aminotransferase(AST), alanine aminotransferase(ALT), total bilirubin(TBIL) and direct bilirubin(DBIL) in serum of rats were determined by a fully automatic biochemical analyzer. The mRNA expression level of miR-199a-5p in liver tissue of rats was detected by qRT-PCR. The protein expression levels of α-smooth muscle actin(α-SMA) and collagen type 1 alpha 1(COL1A1) in liver tissue of rats were detected by double immunofluorescence staining and Western blot experiment. Rat feces were collected for 16S rRNA high-throughput sequencing. Results The expression of miR-199a-5p was up-regulated in the liver tissue of BDL rats(P<0.01). Compared with the NC adv group, the degree of liver injury and collagen deposition were relatively serious, the levels of AST, ALT, TBIL and DBIL in serum and the expression levels of α-SMA and COL1A1 in liver tissue increased in the miR-199a-5p adv group(allP<0.05). However, the results of miR-199a-5p sponge adv intervention were opposite(allP<0.05). The 16S rRNA sequencing results showed that rats treated with miR-199a-5p adv were characterized by increased diversity and richness of intestinal microbiota, changed composition of intestinal microbiota, while the results of miR-199a-5p sponge adv interfering with the bacterial community were opposite(allP<0.05). Conclusion miR-199a-5p promotes liver fibrosis of BDL rats, and its mechanism may be related to regulating the diversity and abundance of intestinal microbiota.

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Objective To explore the mechanisms of circadian rhythm disorder(CRD) on behavior and testicular spermatogenic capacity in adolescent mice. Methods Thirty SPF grade C57 mice were selected and randomly divided into the control and CRD groups with 15 mice in each group. The control group kept 12 h dark/12 h bright circulating light, and the CRD group kept 24 h light. The trial lasted for 61 days. The growth curves of mice in each group were counted; the elevated plus maze test and open field test were performed to detect mice behavior; neuronal morphology was visualized by Nissl staining. The distribution of ionized calcium-binding adapter molecule 1(Iba1) and neuron-specific nuclear protein(NeuN) in the hypothalamus were detected by immunofluorescence and Western blot. Expression of testosterone synthesis-related genes steroidogenic acute regulatory protein(StAR) and hydroxy-delta-5-steroid dehydrogenase, 3 beta-and steroid delta-isomerase 1(HSD3B1) and spermatogenesis-related genes gametogenetin binding protein 2(GGNBP2) and deleted in azoospermia-like(DAZL) were determined by RT-qPCR. Results The weight of the CRD group was significantly higher than that of control group at 61 days; in the elevated plus maze test, the time, frequency, and percentage of time in the open arm of the CRD group were significantly less than those of the control group; in the open field test, there was no significant difference in movement distance between the two groups; however, the residence time of the central area in the CRD group was significantly less than that in the control group; the frequency of entering the central area in the CRD group was significantly less than that in the control group. Nissl staining results showed that the positive cells in the CRD group were significantly lower than the control group. Immunofluorescence and Western blot results showed that Iba1 protein expression was up-regulated and NeuN protein expression was down-regulated in the hypothalamus of the CRD group. In the RT-qPCR experiment, the expression of HSD3B1 in the CRD group was significantly lower than that of the control group; the expression of GGNBP2 and DAZL in the CRD group was significantly lower than that in the control group. Conclusion The CRD treatment can not only lead to depressive behavior in adolescent mice but also reduce the development of reproductive system in male adolescent mice.

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Objective To study the effect of plasma exosomes on hypoxia tolerance of human microvascular endothelial cells(HMEC-1) in stroke patients and healthy controls before and after remote ischemic preconditioning(RIPC) treatment. Methods HMEC-1 cells were divided into a blank control group(Group C) and oxygen glucose deprivation(OGD) treatment group, using plasma exosomes from stroke patients before and after RIPC(S-b group, S-a group) and healthy adults before and after RIPC(C-b group, C-a group). HMEC-1 cells were incubated with a concentration of exosome(35 μg/ml) for 24 hours, followed by OGD treatment for 6 hours and reperfusion treatment for 24 hours in all groups. Water-soluble tetrazolium salt-1(WST-1) method was used to detect endothelial cell viability, caspase-3 enzyme activity was measured, and Western blot was used to detect the expression levels of zone occluden-1(ZO-1) and claudin-5.Results After RIPC in healthy adults, plasma exosomes significantly increased the cell activity of HMEC-1 cells and decreased the activity of caspase-3 enzyme in HMEC-1 cells.Conclusion After RIPC treatment, plasma exosomes reduced endothelial cell apoptosis and increased endothelial cell hypoxia tolerance.

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Objective To explore the changes in cerebral blood flow caused by mandibular retrusion, as well as the impact and potential mechanisms on stroke recovery. Methods 6-week-old SD male rats were selected as experimental subjects. The metal cannula was bonded to the rat maxillary incisor for one week, forcing mandibular retrusion(MR). Cerebral blood flow was detected by laser speckle imaging. Cognitive function was detected by the Morris water. Then, the stroke model was constructed in MR rats by using the middle cerebral artery occlusion(MCAO) method for one week. Meanwhile, metal cannulae were then removed in rats to restore the lower jaw′s position(MCAO RO), serving as a positive control group. Consequently, rats were randomly divided into the following groups: Sham groups, MCAO groups, MCAO MR groups, and MCAO RO groups. Neurological recovery was assessed through the modified neurological severity score(mNSS). The area of cerebral infarction was evaluated by using triphenyltetrazolium(TTC) staining. The changes in nerve cells were observed by using hematoxylin eosin(HE) staining. The protein expression level of vascular endothelial growth factor(VEGF) was detected by immunohistochemistry. The protein expression levels of platelet-endothelial cell adhesion molecule(CD31), sirtuin 6(SIRT6), and thioredoxin interaction protein(TXNIP) were detected by Western blot. The mRNA expression levels of SIRT6, TXNIP, and VEGF were determined by qRT-PCR. Microglia activation marker molecule 1(IBA-1) was detected by immunofluorescence. Resluts Because of mandibular retrusion, laser speckle showed decreased cerebral blood flow, and the water maze showed decreased cognitive function. Compared to other groups, MCAO MR showed a larger ischemic area in TTC staining, while HE staining and neurological scoring showed poorer neurological function recovery. Western blot and qRT-PCR showed that the MCAO MR group inhibited the mRNA and protein expression levels of SIRT6, upregulated the mRNA and protein expression levels of TXNIP, and increased the activation of microglia. Conclusion Mandibular retrusion reduces cerebral blood flow and alters cognitive function in rats. Mandibular retrusion inhibits recovery in stroke through the SIRT6/TXNIP axis.

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Objective To investigate the effect of leflunomide(LEF) on the expression of associated autophagy genes in synoviocytes of rheumatoid arthritis(RA) by regulating HIF-1α signal pathway. Methods Three generations of RA synovial cells were divided into blank control group, LEF group and Tripterygium wilfordii polyglycosides group. The blank control group was added with the same volume of DMEM culture medium. The drug group was treated with LEF(concentration 0.2 mg/ml) and Tripterygium wilfordii polyglycosides(concentration 0.03 mg/ml), the proliferation and apoptosis of synovial cells were detected by flow cytometry, the expression of IL-1β, TNF-α, ANGPTL-4 and VEGF was detected by ELISA, the expression of HIF-1α mRNA was detected by qRT-PCR, and the expression of HIF-1 α, Beclin-1 and BNIP3 protein was detected by Western blot. Results Compared with Tripterygium wilfordii polyglycosides group, the expression of IL-1 α, TNF-α, ANGPTL-4 and VEGF in synovial supernatant of LEF group decreased; compared with the blank control group, the expression of HIF-1α mRNA in synovial cells of LEF group and Tripterygium wilfordii polyglycosides group decreased, and the effect of LEF group was the most obvious; compared with the blank control group, the protein expressions of HIF-1α, Beclin-1 and BNIP3 in synovial cells of LEF group and Tripterygium wilfordii polyglycosides group decreased, and the effect of LEF group was the most obvious. Conclusion LEF can inhibit the expression of inflammatory factors in RA synovial cells, inhibit HIF-1α signaling pathway, inhibit the expression of autophagy-related genes Beclin-1 and BNIP3, and improve the pathological state of synovitis.

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Objective To screen mitochondria-targeting differential genes in alveolar macrophages of silicosis patients and explore the role of mitochondrial homeostasis in alveolar macrophages of silicosis patients. Methods High-throughput sequencing dataset GSE174725 was downloaded, and differentially expressed genes were screened with R software andP<0.05, |LogFC|>1, and then intermixed with mitochondrial gene bank MitoCarta3.0 to obtain mitochondria-targeted differentially expressed genes. Then enrichment analysis was carried out to obtain the biological processes and pathways involved in differential genes, and the protein-protein interaction network was constructed. In addition, alveolar macrophages from silicosis patients and healthy controls were collected, the expression levels of differential genes were detected by RT-qPCR, and the expressions of mitochondria-related factors mitochondrial fusion protein 1(MFN1), optic atrophy 1(OPA1) and dynamin-related protein 1(DRP1) in alveolar macrophages of silicosis patients were investigated by Western blot. Results A total of 204 differentially expressedgenes were screened in silicosis patients, among which 62 differentially expressed genes were up-regulated, 142 differentially expressed genes were down-regulated, and 22 differentially expressed genes were mitochondria-targeted. The concentration analysis of differentially expressed genes targeted by mitochondria showed that the cell components mainly enriched included mitochondrial membrane, endoplasmic membrane side components, etc. The biological processes mainly enriched included mitochondrial electron transfer from NADH to ubiquinone, inflammatory response, immune response, etc. The main molecular functions enriched included the rotation mechanism of proton transport ATP synthase activity, NADH dehydrogenase activity, chemokine activity, etc. KEGG enrichment analysis mainly focused on the involvement in chemical carcinogenesis-ROS, IL-17 signaling pathway, toll-like receptor signaling pathway, chemokine signaling pathway, TNF signaling pathway, etc. In addition, RT-qPCR results showed that the expressions of mitochondrial cytochrome coxidase 1, mitochondrial cytochrome coxidase 2, mitochondrial cytochrome coxidase 3, mitochondrially encoded NADH dehydrogenase 1, mitochondrially encoded NADH dehydrogenase 3, mitochondrially encoded NADH dehydrogenase 5, superoxide dismutase and mitochondrially encoded ATP synthase 6 gene were down-regulated in silicosis patients(P<0.05). Western blot and RT-qPCR results showed that, in silicosis patients, the expression of MFN1 and OPA1 decreased(P<0.05), while the expression of DRP1 increased(P<0.05). Conclusion Bioinformatics analysis and validation, eight mitochondrial targeted differential genes(MT-CO1, MT-CO2, MT-CO3, MT-ND1, MT-ND3, MT-ND5, SOD and MT-ATP6) were finally obtained, which were enriched in mitochondrial respiratory chain and oxidative stress pathways and might play an important role in the process of silicosis.

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Objective To explore the effects of polystyrene microplastics(PS-MPs) on the growth, activity, oxidative stress levels, biofilm formation and virulence ofKlebsiella pneumoniae. Methods Klebsiella pneumoniaewas exposed to PS-MPs at different concentrations(10, 50 and 100 μg/ml) and particle sizes(0.1, 1.0 and 5.0 μm), and the growth curves were measured. The bacterial activity was determined by CCK-8(cell counting kit-8). The level of intracellular reactive oxygen species(ROS) was determined by fluorescence probe. The biofilm forming ability was determined by crystal violet staining. Real-time quantitative fluorescent PCR(qRT-PCR) was used to detect the relative expression levels of biofilm-forming genes(luxS, mrkA, wbbM, pgaA, wzm) and virulence genes(ureA, uge, wabG, fimH). Results A high concentration(100 μg/ml) of 0.1 μm PS-MPs had a stronger inhibitory effect on the growth and activity ofKlebsiella pneumoniae, and the intracellular ROS level significantly increased, indicating that smaller particle size and higher concentration of PS-MPs were more toxic to bacteria. PS-MPs of 100 μg/ml particle size groups(0.1, 1.0 and 5.0 μm) significantly promoted the biofilm formation ofKlebsiella pneumoniae. The relative expression levels of biofilm formation related genes(luxS, mrkA, wbbM, pgaA, wzm) and virulence genes(ureA, uge, wabG, fimH) increased. Conclusion By inducingKlebsiella pneumoniaeto produce a high level of ROS, PS-MPs can cause oxidative stress, inhibit the growth and activity of bacteria, and enhance the biofilm formation ability and virulence, thus affecting the biological characteristics ofKlebsiae pneumoniae.

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Objective To investigate the survival time of patients diagnosed with sporadic Creutzfeldt-Jakob disease in China between 2020 and 2022 and explore the associated factors influencing survival. Methods A retrospective analysis was conducted on clinically diagnosed cases with complete information on sporadic Creutzfeldt-Jakob disease diagnosed by the China Creutzfeldt-Jakob disease surveillance network from 2020 to 2022, baseline information of patients was obtained from the case files, telephone follow-up was used to obtain the treatment and survival status of the patients after the diagnosis, life-table method was used for estimating the survival rate, Kaplan-Meier method was used for calculating the median survival time and the 95%CI, Cox regression model was used for univariate and multivariate analyses were used to screen for factors influencing survival time. Results The median survival time of the 300 patients was 5 months(95%CI: 4.165-5.835). Univariate analysis revealed that factors such as age at onset, regional distribution, presence of corticobasal or extrapyramidal symptoms as initial manifestations, number of initial symptoms, presence of corticobasal or extrapyramidal functional abnormalities, number of major clinical manifestations, presence of typical electroencephalogram findings, and use of nasal feeding during the course of the disease were potential factors influencing survival time(P<0.1). Multivariate analysis showed that the risk of death in patients with onset age>65 years was 1.350 times higher than in patients with onset age ≤65 years(P=0.021, 95.0%CI: 1.046-1.742). Patients without pyramidal or extrapyramidal dysfunction had a 0.674-fold lower risk of death compared to those with these symptoms(P=0.020, 95.0%CI: 0.483-0.939). Patients who did not receive nasal feeding had a 1.817-fold higher risk of death compared to those who did(P<0.001, 95.0%CI: 1.406-2.349). Conclusion Age at onset, the presence of pyramidal or extrapyramidal functional abnormalities, and the use of nasal feeding during the disease course are factors influencing the survival time of patients clinically diagnosed with sCJD.

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Objective To analyze the interaction between voriconazole(VRC) and immunosuppressants such as tacrolimus and cyclosporine and the effect of CYP2C19 gene polymorphism on the interaction and adverse reactions(ADR) based on the results of CYP2C19 gene polymorphism and therapeutic drug monitoring, so as to provide a basis for the development of individualized VRC combined with immunosuppressants. Methods Two-dimensional liquid chromatography and pyrosequencing was used to detect the concentration of VRC and the CYP2C19 gene polymorphism, respectively. And the concentration of immunosuppressants was detected at the same time. The relationship among CYP2C19 gene polymorphism, the concentration of VRC and immunosuppressant and ADR was analyzed. Results A total of 61 patients were enrolled in this study, and the mutation rates of CYP2C19*2 and CYP2C19*3 were 54.1%(33/61) and 9.84%(6/61), respectively. The concentrations of VRC in patients with extensive metabolism(EMs), intermediate metabolism(IMs) and poor metabolism(PMs) were(4.44±3.46),(3.62±3.02) and(10.05±1.46) μg/ml(P<0.05), respectively. The concentration of tacrolimus after combined with VRC significantly increased compared to tacrolimus alone [(13.4±9.2) ng/mlvs(6.5±3.6) ng/ml;P=0.002], and the concentration of tacrolimus increased along with an increasing of VRC concentration. The concentration of VRC in patients combined with tacrolimus was lower than that in patients without immunosuppressants [(3.81±3.48) μg/mlvs(5.84±3.71) μg/ml;P=0.032]. The concentration of VRC in patients with cyclosporine significantly decreased(P<0.01), while tacrolimus and mycophenolate mofetil had no significant effect on the concentration of VRC. 45.90%(28/61) of the patients had adverse reactions, the concentration of VRC in patients with ADR was significantly higher than that in patients without ADR [(7.07±3.43) μg/mlvs(3.06±2.90) μg/ml;P<0.001]. And the concentration of VRC in patients with ADR was higher than patients without ADR with based on CYP2C19 genotype. Conclusion CYP2C19 gene polymorphism can significantly affect the concentration and adverse reactions of VRC, and VRC has significant interaction with immunosuppressants such as tacrolimus. CYP2C19 gene polymorphism combined with therapeutic drug monitoring can improve the individualized treatment of tacrolimus and voriconazole, and is expected to minimize toxicity and improve treatment effects.

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Objective To analyze the causal relationship between Sjogren′s syndrome(SS) and primary biliary cholangitis(PBC) and abnormal liver function by two-sample Mendelian randomization. Methods From the genome-wide association study, single nucleotide polymorphisms of SS, PBC, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, γ-glutamyltransferase, direct bilirubin and total bilirubin levels were extracted as instrumental variables. Inverse-variance weighted(IVW) analysis was used as the main analysis, supplemented by weighted median, MR-Egger, weighted mode, and simple mode. And a series of sensitivity analyses were carried out to verify the robustness of the results. Results The IVW analysis results of PBC on SS were as follows:P=8.57E-11,OR=1.185 9,95%CI=1.126 4-1.248 5;IVW analysis of PBC on SS(P>0.05). IVW analysis results of SS on alkaline phosphatase were as follows:P=0.041 5,Beta=-0.007 2,95%CI=-0.014 0--0.000 3; IVW analysis results of SS on alanine aminotransferase, aspartate aminotransferase, γ-glutamyltransferase, direct bilirubin, total bilirubin levels showed no statistically significant difference. Conclusion PBC can significantly increase the risk of SS, but SS cannot be proved to have a causal effect on the occurrence of PBC. SS has a potential effect on reducing alkaline phosphatase levels, but it is not proved that SS has causal relationship with alanine aminotransferase, aspartate aminotransferase, γ-glutamyltransferase, direct bilirubin, total bilirubin levels.

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Objective To compare the differences in lipid metabolism between platinum-resistant and platinum-sensitive ovarian cancer patients at stage Ⅲ-Ⅳ, to analyze the differential intestinal flora using 16S rRNA sequencing, and to explore the associations among intestinal flora, lipid metabolism characteristics and platinum resistance in ovarian cancer. Methods Patients diagnosed with ovarian cancer at stage Ⅲ-Ⅳ through surgical pathology were selected, including a platinum-resistant group(11 cases) and a platinum-sensitive group(11 cases). The differences in lipid metabolism between the two groups were compared. The differences in gut microbiota between the two groups were investigated using fecal 16S rRNA sequencing. The association among gut microbiota, lipid metabolism characteristics, and platinum resistance in ovarian cancer was analyzed. Results Significant differences were observed in lipid metabolism-related indicators[total cholesterol(TC), high-density lipoprotein cholesterol(HDL-C), non-high-density lipoprotein cholesterol(n-HDL), low-density lipoprotein cholesterol(LDL-C), apolipoprotein(B)] between the two groups, with higher levels in the platinum-resistant group. The Shannon index(P=0.008 3) and Simpson index(P=0.008 2) both showed higher diversity of gut microbiota in platinum-resistant ovarian cancer patients compared to the platinum-sensitive group. However, based on OTUs species clustering and relative abundance statistics, certain bacterial abundances differed significantly between the groups. Species such asParabacteroides,Akkermansia,Blautia,Lachnoclostridium,Fusicatenibacter, andMegamonashad significantly higher abundances in the platinum-sensitive ovarian cancer group, andAkkermansia(a lipid metabolism-related bacterial group) was the most prevalent. Conclusion The platinum-resistant group of ovarian cancer exhibits significantly higher levels of lipid metabolism and gut microbiota diversity compared to the platinum-sensitive group. This suggests that the increase in lipid metabolism levels and fecal microbiota diversity may be associated with the development of platinum resistance. However, certain microbial taxa are reduced in abundance in the platinum-resistant group, such as the distinctAkkermansiagenus(a lipid metabolism-related microbial community), which may serve as one of the factors inducing platinum-resistance in ovarian cancer.

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Objective To investigate the correlation between the appendicular skeletal mass index(ASMI) and osteoporosis(OP) in postmenopausal patients with type 2 diabetes(T2DM). Methods 164 hospitalized postmenopausal elderly T2DM patients were selected and their bone density(BMD)and appendicular skeletal mass were measured using dual energy X-ray absorption method(DXA). ASMI=appendicular skeletal mass/height2(kg/m2), and they were divided into OP group and non OP group based on T value. The general clinical data, blood biochemical indicators, and ASMI between the two groups were compared, and Logistic regression analysis, ROC curve were further used to analyze the correlation and diagnostic power. Results Compared with non OP group, OP group had a higher incidence of sarcopenia(SAC)(P<0.05); the differences in age, ASMI, body mass index(BMI), and estradiol(E2) had significant differences between the two groups(P<0.05); Logistic regression analysis showed that the reduction of ASMI [OR=0.133,95%CI(0.029-0.611)], BMI[OR=0.785,95%CI(0.625-0.985)], and E2 [OR=0.967, 95%CI(0.942-0.993)] were protective factors for OP; receiver operating curve(ROC) suggested that the AUC of ASMI for OP prediction was 0.752 [95%CI(0.632-0.872),P<0.001] with sensitivity 87.5%,specificity 47.8%,and the best diagnostic value was 5.52 kg/m2. Conclusion The reduction of ASMI, BMI, and E2 is positively correlated with the occurrence of osteoporosis. ASMI is an important protective factor for osteoporosis. Early OP screening and risk factor assessment should be conducted for elderly postmenopausal T2DM patients, and early intervention measures should be taken to reduce the risk of falls and fractures.

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Objective To assess the influence of inter-implant distances on the accuracy of intraoral scanning impressionsin vitro. Methods A master cast model with different inter-implant distances was scanned by laboratory and intraoral scanners, which were used as the reference and intraoral scan data. The distance and angular deviations of the scan bodies corresponding to the reference and intraoral digital scan standard tessellation language(STL) files were measured and calculated in a three-dimensional analysis software. The one-samplet-test, Spearman′s correlation analysis,Brown-Forsythe Ftest,Games-Howell post-hoctest, and Levene′s test were used for comparisons(α=0.05). Results The overall distance and angular deviations of the intraoral digital scan were(27.48±18.14) μm and(0.24±0.19) °, within clinically acceptable limits(P<0.001). The inter-implant distances exhibited a significant positive correlation with both the scanning distance and angular deviations in terms of scanning trueness. The angular deviation differed significantly between the 1-2 group(8.13 mm) and the other distance groups. Additionally, inter-implant distances affected the precision of the intraoral scanner(P<0.05). Conclusion The findings of this study indicate that intraoral scanning impressions of complete-arch implant-supported prostheses are within clinically acceptable ranges of accuracy. Inter-implant distances ≤ 8.13 mm can result in a higher accuracy of intraoral scanning. Increased inter-implant distances can adversely affect intraoral scanning accuracy.

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Objective To evaluate the effect of modified vertical tooth preparation with full zirconium crown restoration in molar area. Methods A randomized study was conducted in 84 patients with all-ceramic crown restoration of posterior teeth. 90 full crown restorations were collected and randomly divided into two groups: 45 teeth in experimental group were treated with modified vertical tooth preparation, while 45 teeth in control group were treated with conventional horizontal tooth preparation with full zirconium crown restoration. When patients wore teeth, the restoration effects of the two groups were evaluated respectively, and then the marginal and internal fitness of the two groups were measured by silicone rubber replication method; gingival index(GI) and sulcus bleeding index(SBI) scores were recorded 6 months after wearing teeth. Results The results of chair-side evaluation of the two groups showed that the marginal fitness of the experimental group was better than that of the control group, and the difference was statistically significant(P=0.036); the measured value of marginal fitness of prostheses in experimental group was better than that in control group, and the difference was statistically significant(P<0.001); after 6 months, there was no mechanical complication between the two groups, and there was no significant difference in GI and SBI scores of periodontal soft tissue indexes. Conclusion Modified vertical tooth preparation abutment can significantly improve the marginal fitness of all-ceramic crown, which will not change the health status of peri-crown gingiva.