Objective To develop a method capable of real-time,rapid,and visual detection of MEs,providing a powerful tool for early diagnosis and assessment of diseases.Methods In this study,flake graphene oxide(2D GO)with excellent peroxidase-like activity(POD-like)was prepared by a modified Hummers'method,and a sensing strategy for the rapid visual detection of MEs was constructed by integrating colorimetry with a smartphone.Results 2D GO can catalyse the decomposition of hydrogen peroxide(H2O2)to produce highly oxidative hydroxyl radicals(·OH),which oxidize the colourless 3,3',5,5'-tetramethylbenzidine(TMB)into blue ox-TMB.The strong reducing effect of MEs on the"2D GO+TMB+H2O2"colorimetric sensing system enables rapid visual detection of MEs.Conclusion The colorimetric platform constructed based on 2D GO has a wide linear detection range(10-1 000 μmol/L)and a good detection limit(LOD<7 μM),and was successfully used for the determination of MEs in foetal bovine serum samples,with satisfactory recovery rates.

In this study,the carboxylated magnetic beads were coupled with bivalent nanobodies a-gainst porcine epidemic diarrhea virus(PEDV)M protein to construct immunomagnetic beads(IM-NBs-Ⅱ),The capture and enrichment function of IMNBs-Ⅱ was verified by using PEDV propaga-ted in Vero cells.A one-step RT-qPCR detection method for PEDV was established by combining the characteristics of IMNBs-Ⅱ with the detection advantages of reverse transcription fluorescence quantitative polymerase chain reaction(RT-qPCR).Specific analysis found that this method has no cross reactivity with swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,porcine circovirus,indicating that it has good specificity.Sensitivity analysis re-vealed that the detection sensitivity of the RT-qPCR based on IMNBs-Ⅱ was increased 10 times compared to traditional RT-qPCR methods.Detection of the clinical samples confirm that the RT qPCR method based on IMNBs-Ⅱ is suitable for rapid and accurate detection of clinical feces and tissue samples.The method established in this study effectively avoids contamination issues during nucleic acid extraction,simplifies experimental procedures,and saves detection time,which pro-vides a method for efficient detection of PEDV.

In this study,the carboxylated magnetic beads were coupled with bivalent nanobodies a-gainst porcine epidemic diarrhea virus(PEDV)M protein to construct immunomagnetic beads(IM-NBs-Ⅱ),The capture and enrichment function of IMNBs-Ⅱ was verified by using PEDV propaga-ted in Vero cells.A one-step RT-qPCR detection method for PEDV was established by combining the characteristics of IMNBs-Ⅱ with the detection advantages of reverse transcription fluorescence quantitative polymerase chain reaction(RT-qPCR).Specific analysis found that this method has no cross reactivity with swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,porcine circovirus,indicating that it has good specificity.Sensitivity analysis re-vealed that the detection sensitivity of the RT-qPCR based on IMNBs-Ⅱ was increased 10 times compared to traditional RT-qPCR methods.Detection of the clinical samples confirm that the RT qPCR method based on IMNBs-Ⅱ is suitable for rapid and accurate detection of clinical feces and tissue samples.The method established in this study effectively avoids contamination issues during nucleic acid extraction,simplifies experimental procedures,and saves detection time,which pro-vides a method for efficient detection of PEDV.

ObjectiveTo investigate the regulatory effect of circular RNA circ_0120051 on the fibrotic phenotype of cardiac fibroblasts and the potential mechanism involved. MethodsThe expression of circ_0120051 and its host gene of solute carrier family 8 member A1（SLC8A1） mRNA in the myocardium of healthy organ donors （n=24） and heart failure （HF） patients （n=21） were assessed by real-time quantitative polymerase chain reaction （RT-qPCR） assay. RNA stability of circ_0120051 was identified by RNase R exonuclease digestion assay. The cytoplasmic and nuclear distribution of circ_0120051 in human cardiomyocyte AC16 was detected by RT-qPCR assay. The expression of fibrosis-related genes in mouse cardiac fibroblasts （mCFs） with adenovirus-mediated overexpression of circ_0120051 was detected by RT-qPCR and Western blot assay， respectively. The effect of overexpression of circ_0120051 on the migration activity of mCFs was evaluated by wound-healing assay. RNA co-immunoprecipitation （RIP） was conducted to detect the interaction between circ_0120051 and miR-144-3p. The binding site of miR-144-3p in the 3'-UTR of isocitrate dehydrogenase 2 （Idh2） mRNA was identified by the dual luciferase reporter gene assay. ResultsCirc_0120051 was significantly up-regulated in the myocardium of HF patients， while the mRNA expression of its host gene SLC8A1 was not changed. Circ_0120051 was mainly located in the cytoplasm of human AC16 cells. Results of RNase R exonuclease digestion revealed that circ_0120051 possesses the characteristic stability of circular RNA compared to the linear SLC8A1 mRNA. Overexpression of circ_0120051 could inhibit the expression of fibrosis-related gene in mCFs and mCFs migration. RIP assay confirmed the specific interaction between circ_0120051 and miR-144-3p. Transfection of miR-144-3p mimic could efficiently promote the expression of fibrosis-related genes in mCFs and reverse the inhibitory effect of circ_0120051 on the fibrotic phenotype of mCFs. Results of the dual luciferase reporter gene assay confirmed the interaction between miR-144-3p and the 3'-UTR of Idh2. Transfection of miR-144-3p transcriptionally inhibited Idh2 expression， and overexpression of circ_0120051 enhanced IDH2 expression in mCFs. MiR-144-3p mimic and Idh2 small interfering RNA （siRNA） could consistently reverse the inhibitory effects of circ_0120051 on fibrosis-related genes expression in mCFs and mCFs migration. ConclusionsCirc_0120051 inhibits the fibrotic phenotype of cardiac fibroblasts via sponging miR-144-3p to enhance the target gene of IDH2 expression.

Hypoxia-inducible factor (HIF-1α), a core transcription factor responding to changes in cellular oxygen levels, is closely associated with a wide range of physiological and pathological conditions. However, its differential impacts on vascular cell types and molecular programs modulating human vascular homeostasis and regeneration remain largely elusive. Here, we applied CRISPR/Cas9-mediated gene editing of human embryonic stem cells and directed differentiation to generate HIF-1α-deficient human vascular cells including vascular endothelial cells, vascular smooth muscle cells, and mesenchymal stem cells (MSCs), as a platform for discovering cell type-specific hypoxia-induced response mechanisms. Through comparative molecular profiling across cell types under normoxic and hypoxic conditions, we provide insight into the indispensable role of HIF-1α in the promotion of ischemic vascular regeneration. We found human MSCs to be the vascular cell type most susceptible to HIF-1α deficiency, and that transcriptional inactivation of ANKZF1, an effector of HIF-1α, impaired pro-angiogenic processes. Altogether, our findings deepen the understanding of HIF-1α in human angiogenesis and support further explorations of novel therapeutic strategies of vascular regeneration against ischemic damage.
Humans ; Vascular Endothelial Growth Factor A/metabolism* ; Endothelial Cells/metabolism* ; Transcription Factors/metabolism* ; Gene Expression Regulation ; Hypoxia/metabolism* ; Cell Hypoxia/physiology*

OBJECTIVE To provide reference for constructing scientific and reasonable pharmaceutical service mode for hospitalized patients with chronic airway diseases. METHODS From October 2023 to March 2024， 250 patients with chronic obstructive pulmonary disease and acute exacerbation of asthma who were hospitalized in the respiratory department of Hebei General Hospital and received pharmaceutical care （PC） were randomly divided into control group （125 cases） and observation group （125 cases）. The control group received general pharmaceutical services throughout their hospitalization， while the observation group received standardized and graded pharmaceutical services throughout their hospitalization. The differences in clinical value indicators， humanistic value indicators， and quality management indicators were compared among different PC service models. RESULTS Among clinical value evaluation indicators， the observation group had better achievement rate of disease treatment goals， correct use score of inhalation devices， the incidence of adverse drug reactions， and the number of drug-related problems solved than the control group （P＜0.05）. Among the humanistic evaluation indicators， compared with the control group， the observation group had better medication compliance scores， pharmacist intervention success rates， and patient satisfaction scores （P＜0.05）. Among quality management evaluation indicators， the proportion of drug costs， the proportion of intravenous medication， the use rate of antibiotics， the intensity of antibiotic use， and the number of pharmaceutical services in the observation group were significantly better than the control group （P＜0.05）. CONCLUSIONS Standardized and graded pharmaceutical care services have improved the efficiency of pharmacists and service effectiveness， making it a new pharmaceutical service model worth promoting.

AIM:To investigate the effect of circular RNA MYO9A-006(circMYO9A_006)on hypertrophic phenotype of cardiomyocytes and the underlying mechanism.METHODS:The effect of adenovirus-mediated overexpres-sion of circMYO9A_006 on the expression of hypertrophy-related proteins,including β-myosin heavy chain(β-MHC),skeletal muscle actin alpha 1(ACTA1)and atrial natriuretic peptide(ANP),was evaluated in neonatal mouse ventricular cardiomyocytes(NMVCs).Moreover,a neonatal rat ventricular cardiomyocyte(NRVC)model of phenylephrine(PE)-in-duced hypertrophy was established.The effect of circMYO9A_006 overexpression on NRVC size was ascertained using Phalloidin-iFluor 647 staining method.Dual-luciferase reporter assay was employed to measure the activity of potential in-ternal ribosome entry sites(IRES)in circMYO9A_006.The translation and intracellular location of the circMYO9A_006-translated protein,MYO9A-208aa,were verified using Western blot.To investigate the role of MYO9A-208aa in the ef-fect of circMYO9A_006 on the cardiomyocyte hypertrophic phenotype,we prepared and used the following adenoviruses:the recombinant circMYO9A_006-ORF adenovirus to express MYO9A-208aa,the recombinant circMYO9A_006-ATG-mut adenovirus that does not express MYO9A-208aa,the recombinant circMYO9A_006 adenovirus,and the adenovirus vector control.These were then employed to infect NRVCs.RESULTS:Successful adenovirus-mediated overexpression of circMYO9A_006 was observed in NMVCs.The increased expression of circMYO9A_006 notably reduced the expres-sion of hypertrophy-related proteins in NMVCs(P<0.01).Concurrently,overexpression of circMYO9A_006 substantially reduced the expression of hypertrophy-associated proteins and diminished the size of PE-induced NRVCs(P<0.05).Dual-luciferase reporter assay identified the activity of 2 IRES in circMYO9A_006.Western blot results indicated that circ-MYO9A_006 could produce the MYO9A-208aa protein with an anticipated molecular weight of 28 kD in NRVCs,primari-ly found in the cytoplasm.Elevated expression of both circMYO9A_006 and MYO9A-208aa consistently reduced the ex-pression of hypertrophy-associated proteins(P<0.01),and counteracted the enlarged size of PE-induced NRVCs(P<0.05).However,increased expression of circMYO9A_006-ATG-mut did not counteract the PE-induced hypertrophic phe-notype of NRVCs.CONCLUSION:circMYO9A_006 attenuates the hypertrophic phenotype of cardiomyocytes by synthe-sizing the MYO9A-208aa protein.

Background:Duodenal neuroendocrine tumors(DNETs)are rare tumors,their disease characteristics are currently not well understood.At present,there are no research data on non-ampullary region DNETs in China.Aims:To analyze the clinical characteristics of patients with non-ampullary region DNETs in order to guide clinical practice.Methods:The clinical data ofnon-ampullary region DNETs diagnosed in the First Affiliated Hospital of Air Force Military Medical University from June 2011 to July 2022 were collected.Using the method of retrospective study,analyze the clinical characteristics of patients.Results:Twenty-two non-ampullary region DNETs patients were screened.Among them,8(36.4%)patients'tumor diameter<2 cm,14(63.6%)patients'tumor diameter≥2 cm.When non-ampullary region DNETs were diagnosed,the main clinical symptoms were abdominal distension(59.1%),followed by abdominal pain(41.0%).When diagnosed,half(50%)of patients with non-ampullary DNETs with tumor diameter<2 cm have no clinical symptoms.The clinical symptoms of non-ampullary DNETs patients with tumor diameter≥2 cm were mainly abdominal distension(85.7%),followed by abdominal pain(57.1%),and a few(14.3%)patients had no clinical symptoms.After diagnosed,the survival time of patients with tumor diameter<2 cm was longer than that of patients with tumor diameter≥2 cm(P=0.048).By the end of follow-up,the median survival time of patients with non-ampullary region DNETs was 451.0 months.Six patients had died,all of their tumor diameter were≥2 cm at diagnosis.Three of patients who died had stage Ⅳ at diagnosis,and all had liver metastases.Patients with tumor diameter<2 cm underwent surgical treatment and all survived after surgery.Conclusions:Abdominal distension is the main clinical manifestation of non-ampullary region DNETs patients,and the organ that is more likely to metastasize is the liver.The survival time of patients with non-ampullary region DNETs with tumor diameter<2 cm was longer than that of patients with tumor diameter≥2 cm.

Pinealoma often comorbid with obstructive hydrocephalus due to their specific location and type, therefore, patients with pinealoma need to be treated by a combined ventriculoperitoneal shunt (VPS) along with surgical resection. Shunt failure is the most common complication after surgery. In this paper, we report a case of obstructive hydrocephalus caused by shunt obstruction after pineal gland tumor resection combined with ventriculoperitoneal shunt. The patient first showed low mood, decreased interest and energy, and gradually developed into manifestations of less eating, less talking, less movement, repeated psychiatric visits, and showed no improvement after treatment with antidepressants. The purpose of the current study is to increase the knowledge of these diseases and reduce misdiagnosis.

Pinealoma often comorbid with obstructive hydrocephalus due to their specific location and type, therefore, patients with pinealoma need to be treated by a combined ventriculoperitoneal shunt (VPS) along with surgical resection. Shunt failure is the most common complication after surgery. In this paper, we report a case of obstructive hydrocephalus caused by shunt obstruction after pineal gland tumor resection combined with ventriculoperitoneal shunt. The patient first showed low mood, decreased interest and energy, and gradually developed into manifestations of less eating, less talking, less movement, repeated psychiatric visits, and showed no improvement after treatment with antidepressants. The purpose of the current study is to increase the knowledge of these diseases and reduce misdiagnosis.