Objective To analyze the effect of miR-133a-3p and its potential target transforming growth factor-β-induced factor 2(TGIF2)on the apoptosis and mitochondrial function of acute myeloid leukemia cells OCI-AML3. Methods OCI-AML3 cells were transfected with miR-133a-3p mimic or/and pcDNA-TGIF2.Transfection efficiency was detected by RT-qPCR.The target sites of miR-133a-3p and TGIF2 were predicted by bioinformatics,and the targeting relationship was verified by dual-luciferase reporter gene assay.Cell proliferation was detected by CCK-8 assay and EdU staining.Flow cytometry was applied to detect the apoptosis and mitochondrial membrane potential.ROS level was measured by associated kit.Western blot was used to detect the expression levels of Caspase-3,Caspase-9,TGIF2,Bcl-2,Bax and c-Myc proteins. Results In OCI-AML3 cells,the miR-133a-3p expression was negatively correlated with TGIF2 expression,and miR-133a-3p targetly binded with TGIF2.Compared with those in the control group,the cell viability in the TGIF2 group was significantly enhanced(P<0.05),the number of EdU positive cells was increased significantly,while the apoptosis rate was significantly reduced(P<0.05);the expressions of Bax/Bcl-2,Caspase-3 and Caspase-9 were significantly down-regulated(P<0.05),while the expression level of c-Myc was significantly up-regulated(P<0.05);the mitochondrial membrane potential showed no significant change,whereas ROS content demonstrated a marked decrease(P<0.05).The overexpression of miR-133a-3p reversed the above results.Compared with those in the TGIF2 group,the mimic+TGIF2 group exhibited significantly reduced cell viability(P<0.05),the number of EdU positive cells was significantly declined,and the apoptosis rate was significantly increased(P<0.05);The expression levels of Bax/Bcl-2,Caspase-3 and Caspase-9 were significantly up-regulated(P<0.05),while the expression level of C-MYC was significantly down-regulated(P<0.05);the mitochondrial membrane potential was significantly changed,and ROS level was significantly increased(P<0.05). Conclusion miR-133a-3p inhibits OCI-AML3 cell viability and proliferation,up-regulates ROS level,induces mitochondrial membrane potential depolarization,and promotes apoptosis by targeting TGIF2.

Objective To explore the influence of miR-370-3p on migration and invasion of hepatocellular carcinoma HepG2 cells by targeting STAT3. Methods The miR-NC or miR-370-3p mimic and pcDNA or pcDNA-STAT3 were transfected into HepG2 cells,and the mRNA relative expression levels of miR-370-3p and STAT3 were detected.miR-370-3p mimic and STAT3-wild type(wt)or STAT3-mutant(mut)were co-transfected into HepG2 cells.Finally,the control group,miR-370-3p group,STAT3 group and miR-370-3p+STAT3 group were established.Transwell assay and Wound-healing assay were applied to detect the invasion,migration of cells.Western blot assay was applied to detect the expression level of epithelial-mesenchymal transition associated proteins.The orthotopic xenograft mouse model was established,and mice were divided into miR-NC group and miR-370-3p group. Results Compared with the control group,the relative expression level of miR-370-3p was significantly upregulated in the miR-370-3p mimic group.In the pcDNA-STAT3 group,the relative expression level of STAT3 mRNA was significantly upregulated.Overexpression of miR-370-3p inhibits invasion,migration,and affect the expression of epithelial-mesenchymal transition associated pproteins in HepG2 liver cancer cells.Additionally,miR-370-3p targets STAT3 to suppress the aforementioned metastatic capabilities of HepG2 liver cancer cells.In in vivo experiments,overexpression of miR-370-3p inhibits tumor growth. Conclusion miR-370-3p can inhibit the invasion,migration of hepatocellular carcinoma cells HepG2 by targeting STAT3.

Objective To explore the key role of diaphanous-related formin 1(Diaph1)gene in the kidneys of nephropathy model mice. Methods Wild-type(WT),heterozygous(Diaph1^+/-),and homozygous(Diaph1^-/-)mice were used to establish the control group and the adriamycin model group,respectively,with 7 mice in each group.Mice in the adriamycin model group received adriamycin via tail vein injection,while those in the control group were administered an equal volume of normal saline.The Diaph1 gene was globally knocked out in mice using the CRISPR/Cas9 system,and homozygous mice were obtained by breeding heterozygous littermates.Genotyping was performed by polymerase chain reaction(PCR).Hematoxylin and eosin(HE)staining was used to observe renal morphology in each group.Immunofluorescence staining was applied to detect the expression of Diaph1 in glomeruli and surrounding renal tubules.Serum and urinary albumin levels were measured using a microplate reader method,and creatinine levels were determined by the sarcosine oxidase method.Western blotting was performed to assess Diaph1 protein expression in the renal cortex.Quantitative real-time polymerase chain reaction(qRT-PCR)was conducted to detect the mRNA expression levels of Podocin,CD2-associated protein(Cd2ap),Nephrin,and kin of IRRE-like protein 1(Neph1)in mouse kidneys. Results In wild-type mice,compared with the control group,the adriamycin model group showed significantly decreased body weight(P<0.05)and narrowed glomerular capsule spaces.Biochemical assays revealed elevated urinary albumin(P<0.05)and serum creatinine(P<0.05),along with reduced urinary creatinine(P<0.05).At the molecular level,both mRNA and protein expression of Diaph1 were significantly upregulated in the adriamycin model group(P<0.05).In the Diaph1-knockout mice,compared with adriamycin-treated wild-type mice,adriamycin-treated heterozygous(Diaph1^+/-)and homozygous(Diaph1^-/-)mice exhibited aggravated glomerulosclerosis and further narrowed glomerular capsule spaces.Moreover,renal mRNA expression of Podocin,Cd2ap,and Nephrin was significantly decreased in adriamycin-treated Diaph1^-/- mice(P<0.05),and biochemical analysis showed significantly reduced serum albumin and serum creatinine levels in these mice(P<0.05). Conclusion The Diaph1 gene exerts a critical protective role in renal podocytes and tubular epithelial cells,suggesting that Diaph1 is a potential functional gene in the adriamycin-induced nephropathy model.This provides new insights and directions for the treatment and future research of nephrotic syndrome.

Objective To investigate the effect of small molecule interfering RNA(siRNA)on the proliferation,apoptosis,and programmed cell death-ligand 1(PD-L1)expression of gastric cancer(GC)MGC-803 cells by targeting hypoxia inducible factor-1α(HIF-1α). Methods Cancer and adjacent normal tissues were collected from 35 GC patients.The mRNA expression of patients HIF-1α and PD-L1 was detected,and their correlation was further analyzed.HIF-1α mRNA expression was examined in different GC cell lines,and MGC-803 cells were selected for subsequent experiments.MGC-803 cells were divided into the following groups:normal group,hypoxia group,hypoxia+si-NC group,hypoxia+si-HIF-1α group,hypoxia+si-HIF-1α+pcDNA group,and hypoxia+si-HIF-1α+pcDNA-PD-L1 group.Cell proliferation,apoptosis and related protein expression were assessed.A nude mouse xenograft model was established to verify in vivo effects. Results Compared with those in the adjacent normal tissues,the HIF-1α and PD-L1 mRNA expression levels in the GC tissues were increased,and they were positively correlated(P<0.05).The expression levels of HIF-1α mRNA and protein were higher in MKN-45,SGC-7901,AGS,HGC-27,and MGC-803 cells than in GES-1 cells(P<0.05),with MGC-803 exhibiting the highest expression levels.Silencing HIF-1α downregulated PD-L1 expression in MGC-803 cells under hypoxic conditions,reduced proliferation activity and the expression of proliferating cell nuclear antigen and Bcl-2,and increased apoptosis rate and the expression of Bax and Cleaved-Caspase-3(P<0.05).Silencing HIF-1α reduced xenograft tumor volume,weight,and PD-L1 positive expression rate in nude mice(P<0.05).Overexpression of PD-L1 reversed the above effects. Conclusion Knockdown of HIF-1α expression under hypoxic conditions can downregulate PD-L1 expression,inhibit GC cell proliferation,and promote cell apoptosis.

Objective To investigate the effects of miR-429 on the healing of chronic non-healing wounds and the Wnt/β-catenin signaling pathway. Methods A total of 60 rats were randomly divided into 5 groups:blank control group,model group,negative control group(injected with antagomir NC),LV-429 group(injected with miR-429 antagomir lentivirus),and inhibitor group(injected with miR-429 antagomir lentivirus and Wnt/β-catenin signaling pathway inhibitor IWP-2).The wound healing rates in rats were assessed,and wound tissue samples were collected for measurement of miR-429 levels,as well as the protein expression of Wnt3a and β-catenin,and the expression of vascular endothelial growth factor A(VEGFA)and endothelial nitric oxide synthase(ENOS).Rat serum samples were collectedor measurement of the levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α). Results Compared with that in the model group and the negative control group,the miR-429 expression level was reduced in the LV-429 group and the inhibitor group.Compared with the model group and the negative control group,in the LV-429 group,the wound healing rates,Wnt3a,β-catenin protein expression,and the positive expression rates of VEGFA and ENOS were all increased,while the IL-6 and TNF-α levels were both decreased(P<0.05).In the inhibitor group,the wound healing rate,Wnt3a and β-catenin protein expression,and the positive expression rates of VEGFA and ENOS were all decreased,while the IL-6 and TNF-α levels were both increased(P<0.05). Conclusion Inhibiting the expression of miR-429 can promote the repair of chronic refractory wounds,and this process is mediated by activating the Wnt/β-catenin signaling pathway.

Objective The effect of unc-51 like kinase 1(ULK1)on the invasion and migration of trophoblastic cells in preeclampsia rats was analyzed based on the Wnt signaling pathway. Methods After establishing a preeclampsia rat model,trophoblastic cells were isolated from placental tissues.Preeclampsia trophoblastic cells were randomly divided into 4 groups:negative control group,siRNA ULK1 group,SKL2001 group,and siRNA ULK1+KYA1797K group.The proliferation activity of trophoblastic cells was detected by CCK8 assay;the invasion and migration abilities of trophoblastic cells were detected by Transwell assay;the apoptosis rate of trophoblastic cells was detected by flow cytometry;the expression levels of ULK1 and Wnt pathway proteins in trophoblastic cells were analyzed by Western blotting. Results The expression level of ULK1 in trophoblastic cells of the siRNA ULK1 group and the siRNA ULK1+KYA1797K group was significantly decreased,while the expression level of ULK1 in the SKL2001 group showed no significant change.The proliferation activity of trophoblastic cells in the siRNA ULK1 and SKL2001 groups was increased,their invasion and migration abilities were enhanced,the apoptosis rate was decreased,and the protein expression levels of Wnt3α,β-catenin,c-Myc,and Cyclin D1 were upregulated.In the siRNA ULK1+KYA1797K group,the proliferation activity,invasion and migration abilities,apoptosis rate,and protein expression levels of Wnt3α,β-catenin,c-Myc,and Cyclin D1 showed no significant changes compared with those in the the negative control group. Conclusion Inhibition of ULK1 expression increases the proliferation,migration and invasion activities of trophoblastic cells in preeclampsia rats and reduces the apoptosis rate,and these processes are related to the activation of the Wnt/β-catenin pathway.

Objective To investigate the effects of suppressor of cytokine signaling 1(SOCS-1)on A549 cells infected with Klebsiella pneumoniae via the NOD-like receptor thermal protein domain-associated protein 3(NLRP3)/apoptosis-associated speck-like protein containing a CARD(ASC)/caspase-1 pathway. Methods Human type Ⅱ alveolar epithelial cells(A549 cells)were divided into four experimental groups:siRNA NC group,siRNA SOCS-1 group,pcDNA3.1 NC group,and pcDNA3.1 SOCS-1 group.Three hours after infection with Klebsiella pneumoniae,the corresponding A549 cells were treated with siRNA NC(50 nmol/L),siRNA SOCS-1(50 nmol/L),pcDNA3.1 NC(20 nmol/L),or pcDNA3.1 SOCS-1(20 nmol/L)for 48 h.MTT assays and BrdU staining were used to analyze the proliferative activity of A549 cells;flow cytometry was used to analyze the apoptosis rate of A549 cells;ELISA was used to determine the levels of inflammatory factors in the culture medium of A549 cells in each group;and Western blot analysis was used to detect the expression levels of SOCS-1,NLRP3,ASC,and caspase-1 proteins in A549 cells. Results Infection with Klebsiella pneumoniae led to increased SOCS-1 expression in A549 cells.Compared with the siRNA NC group,the siRNA SOCS-1 group exhibited increased A549 cell proliferation,reduced apoptosis rates and inflammatory cytokine levels,and decreased expression of NLRP3,ASC,and caspase-1.Compared with the pcDNA3.1 NC group,the pcDNA3.1 SOCS-1 group exhibited reduced A549 cell proliferation,increased apoptosis rates and levels of inflammatory factors,and elevated expression levels of NLRP3,ASC,and caspase-1. Conclusion SOCS-1 promotes apoptosis and the release of inflammatory factors in A549 cells infected with Klebsiella pneumoniae and inhibits A549 cell proliferation;this process is closely related to SOCS-1 regulation of the NLRP3/ASC/Caspase-1 signaling pathway.

Objective To investigate the effects of interfering with the expression of the lncRNA NCK1-AS1 on the growth and invasion of the thyroid cancer cell line CAL-62. Methods lncRNA NCK1-AS1 expression in CAL-62 cells was silenced by transfecting the corresponding shRNA.The expression level of lncRNA NCK1-AS1 was detected by RT-PCR.Cell proliferation was assessed by EdU staining,and apoptosis was analyzed by flow cytometry.RT-PCR was used to detect the mRNA expression levels of proliferating cell nuclear antigen(PCNA),p21,and Survivin.Cell invasion was assessed using the Transwell assay,and the number of microtubule-like structures was measured via a microtubule formation assay.Western blot analysis was performed to detect the expression levels of vascular endothelial growth factor(VEGF),E-cadherin,and N-cadherin. Results Compared with the control group,the lncRNA NCK1-AS1 interference group exhibited reduced cell proliferation,increased apoptosis,downregulated PCNA and Survivin mRNA levels,upregulated P21 mRNA levels,decreased number of invading cells and microtubule-like structures,downregulated VEGF and N-cadherin protein expression,and upregulated E-cadherin expression.These differences were statistically significant(P<0.05). Conclusion Silencing the expression of lncRNA NCK1-AS1 inhibits the growth and invasion of CAL-62 thyroid cancer cells.

Objective To analyze the effect of paeonol on spatial learning,memory and depression-like behavior in rats with chronic unpredictable mild stress(CUMS)-induced depression,and to explore its mechanism. Methods The CUMS-induced depression rats were divided into 5 groups:depression group,low-dose,medium-dose and high-dose paeonol groups and Flu group,and the control group was set up.Drug intervention was administered during the third week of stress and continued for one week;rats in the control group received no treatment.The Morris water maze,sucrose preference test,forced swimming test(FST)and tail suspension test(TST)were conducted.The levels of serum CRH,ACTH and CORT,and the expression levels of ERK/CREB proteins in hippocampus were detected. Results Compared with those in the control group,the sucrose preference of rats in the depression group was declined(P<0.05),the escape latency was prolonged,the times of platform crossings and residence time in the third quadrant were decreased(P<0.05),the immobility time in the FST and TST was increased(P<0.05),the levels of serum CRH,ACTH and CORT were increased(P<0.05),and ratios of p-ERK/ERK and p-CREB/CREB in the hippocampus were decreased(P<0.05).Compared with those in the depression group,the above indexes were significantly reversed in rats of the medium- and high-dose paeonol groups and the Flu group(P<0.05). Conclusion Paeonol can improve spatial learning,memory ability and depression-like behavior in rats with CUMS-induced depression,and the mechanism is related to regulation of HPA and ERK/CREB signaling pathways.

Objective To explore the effects of total glucosides of paeony(TGP)on oxidative stress and lung fibrosis in rats with paraquat(PQ)-induced lung injury. Methods Intraperitoneal injection of PQ was performed to induce lung injury in rats,then those rats were intervened with different doses of TGP.The pathological changes of lung tissues,oxidative stress,levels of inflammatory factors,mRNA levels of fibrosis-related factors and expressions of TGF-β1/Smad3 signaling pathway proteins were observed.RLE-6TN cells were treated with PQ and intervened with TGP.The expression levels of TGF-β1/Smad3 pathway related factors in cells were detected. Results After TGP intervention,the lung injury,the levels of MDA and HYP,the levels of serum TNF-α,IL-6 and IL-1β,the mRNA expression levels of α-SMA,Collagen Ⅲ and TIMP-1,and the expression levels of TGF-β1 and p-Smad3/Smad3 were decreased,while the levels of SOD and CAT,and the ratio of p-Smad7/Smad7 were increased,in a dose-dependent manner.After TGP intervention,the expression level of p-Smad7 in RLE-6TN cells was increased,while the expression levels of TGF-β1 and p-Smad3 were decreased. Conclusion TGP can improve oxidative stress and lung fibrosis in PQ rats.The mechanism may be related to TGF-β1/Smad3 signaling pathways.

Objective To explore the effect of rifampicin on miR-28-5p/nuclear factor-erythroid 2-related factor 2(Nrf2)signaling and liver cell injury. Methods L02 cells were divided into 6 groups:control group,low-,medium- ,and high-dose rifampicin groups,miR NC group,and miR-28-5p inhibitor group.The expression level of miR-28-5p,cell viability,apoptosis rate,inflammatory factor levels,superoxide dismutase(SOD),malondialdehyde(MDA),aspartate aminotransferase(AST),and alanine aminotransferase(ALT)in each group of L02 cells were detected.Meanwhile,the targeting relationship between miR-28-5p and Nrf2 in L02 cells was analyzed. Results The expression level of miR-28-5p in L02 cells were increased with the increase of rifampicin concentration.Compared with those in the control group,the viability of L02 cells in the low-,medium-,and high-dose rifampicin groups,miR NC group,and miR-28-5p inhibitor group was decreased,the apoptosis rate was increased,the levels of inflammatory factors,MDA,AST,and ALT were increased,and the levels of SOD and Nrf2 were decreased.Compared with those in the miR NC group,the viability of L02 cells in the miR-28-5p inhibitor group was increased,the apoptosis rate was decreased,the levels of inflammatory factors,MDA,AST,and ALT were decreased,and the levels of SOD and Nrf2 were increased.miR-28-5p could targetly inhibit the expression of Nrf2. Conclusion Rifampicin can promote liver cell injury by miR-28-5p and inhibition of Nrf2.

Objective To explore the effect of ropivacaine on alleviating ovalbumin(OVA)-induced allergic rhinitis(AR)in mice. Methods Mice were randomly divided into 5 groups:control group,AR group,low-dose Ropivacaine group(1 mg/kg),high-dose Ropivacaine group(5 mg/kg)and Dexamethasone(Dex)positive group(2 mg/kg),with 5 mice in each group.Indicator evaluations were conducted after the last drug administration. Results The body weight of mice in the high-dose Ropivacaine group and Dex group was greater than that in the AR group(P<0.05),and the relative spleen weight,the times of nose rubbing,sneezing and runny nose,levels of serum histamine,OVA-specific immunoglobulin E(IgE),total IgE,total IgG1 and β-hexosaminidase,levels of iIL-1β,IL-6,TNF-α,IL-4 and IL-5 in nasal lavage fluid and the protein expression levels of p65 and p-IkBα in the nasal mucosa were lower than those in the AR group(P<0.05).Eosinophil infiltration was reduced. Conclusion High-dose ropivacaine can relieve OVA-induced rhinitis symptoms and inflammatory infiltration of nasal mucosa of mice with AR.The mechanism may be related to the inhibition of NF-κB signaling pathway.

Objective To investigate the therapeutic effect of Mongolian medicine Jixiang Ankun Pill on premature ovarian failure(POF)in rats and explore its potential mechanism. Methods A total of 54 SD female rats were randomly divided into six groups.Except for the control group,the remaining groups were induced to develop a POF model using Tripterygium glycoside tablets.Subsequently,each treatment group was administered the corresponding drug via oral gavage for 36 days.Serum hormone levels,inflammatory factors,and oxidative stress markers were measured;ovarian histopathology was examined;and the expression of molecules related to the PI3K/AKT/mTOR pathway was assessed. Results The model group exhibited hormonal imbalances,abnormal levels of inflammation and oxidative stress,severe damage to ovarian structure,and abnormal expression of PI3K/AKT/mTOR pathway-related molecules.Improvements were observed in all Mongolian medicine concentration groups and the Western medicine group,with the highest concentration group and the Western medicine group showing the most significant effects. Conclusion Jixiang Ankun Pill can effectively improve ovarian function in POF rats via regulating hormone levels,inhibiting inflammation and oxidative stress,and its mechanism is closely related to the regulation of the PI3K/AKT/mTOR signaling pathway and mTOR phosphorylation.

As a key member of the serine/threonine protein kinase family,Polo-like kinase 1(Plk1)is a core regulator of the cell cycle,whose dysfunction is closely related to tumorigenesis.Recent research has found that Plk1 is abnormally expressed in terminally differentiated neurons and is closely associated with the pathological processes of neurodegenerative diseases such as Alzheimer’s disease,Parkinson’s disease and amyotrophic lateral sclerosis.It is particularly noteworthy that Plk1 plays a dual role in diseases like Alzheimer’s:it not only directly mediates neuronal death by driving aberrant cell cycle re-entry but also participates in regulating neuroprotective processes such as autophagy.The complex role of Plk1 in neurodegenerative diseases,particularly its seemingly contradictory dual function,offers a new perspective for understanding the mechanisms underlying neurodegenerative disorders and developing novel therapeutic strategies.

As an innovative high-throughput sequencing technology,nanopore sequencing has become a vital tool in genomics research and clinical diagnostics due to its unique advantages of real-time detection,long-read capability,and PCR-free amplification.In recent years,significant advancements in sequencing speed and data accuracy have driven progress in genome sequencing,transcriptome analysis,and multi-omics integration studies.However,nanopore sequencing still faces challenges such as signal noise,complex data processing,and accuracy improvement.This article systematically reviews the fundamental principles and technological evolution of nanopore sequencing,with a focus on its latest advancements in genome sequencing,transcriptome analysis,and clinical applications.In addition,by exploring current technical bottlenecks and future development directions,this article aims to provide comprehensive theoretical support and practical guidance for researchers,thereby promoting the widespread application of nanopore sequencing technology in life sciences and medicine.