Positive response of a hemodialysis patient with pure red cell aplasia on recombinant human erythropoietin therapy to cyclosporine and Roxadustat.

Recombinant human erythropoietin (rHuEPO) is commonly used to treat anemia associated with chronic kidney disease (CKD). EPO-induced Pure Red Cell Aplasia (PRCA) is a rare condition of profound anemia with EPO treatment. Upon finding the development of EPO-induced PRCA, the treatment requires immediate withdrawal of EPO therapy and initiate new treatments with immunosuppression or renal transplantation. Anti-EPO antibody assay is not always positive in EPO-induced PRCA. Here, we report a case on the sudden development of PRCA in a hemodialysis patient receiving EPO and how we treated the condition successfully with cyclosporine and subsequently maintained the hemoglobin with Roxadustat, a hypoxia-inducible factor prolyl hydroxylase inhibitor (HIF-PHI). Even though the anti-EPO antibody was negative by Enzyme Linked Immunosorbent Assay (ELISA) in our case, the clinical course, the markedly reduced reticulocyte count < 10,000/µL, the bone marrow (BM) biopsy revealing reduced erythroblasts, and its subsequent response to cyclosporine, were similar to EPO-induced PRCA. The clinical picture of EPO-induced PRCA, the limitation of the EPO-neutralizing antibody (Ab) assay, and treatment strategies were discussed.

Principle and design of clinical efficacy observation of extracorporeal cardiac shock wave therapy for patients with myocardial ischemia-reperfusion injury: A prospective randomized controlled trial protocol.

BACKGROUND: Acute ST-segment elevation myocardial infarction (STEMI) remains a serious life threatening event with a poor prognosis due to myocardial ischemia/reperfusion injury despite coronary revascularization. Extracorporeal cardiac shock wave (ECSW) is a safe, effective and non-invasive new method for the treatment of cardiovascular diseases. The current results show that extracorporeal cardiac shock wave provides a new treatment option for patients with severe and advanced coronary heart disease. However, there are relatively few clinical studies on the application of in vitro cardiac shock waves in patients with myocardial ischemia-reperfusion injury. We hypothesized that extracorporeal cardiac shock therapy would also be effective in reducing clinical endpoints in patients with STEMI reperfusion. OBJECTIVE: This study is order to provide a new therapeutic method for patients with myocardial ischemia-reperfusion injury and reveal the possible mechanism of ECSW for ischemia-reperfusion injury. METHODS AND MATERIALS: CEECSWIIRI is a single-center, prospective randomized controlled trial that plans to enroll 102 eligible patients with acute ST-segment elevation myocardial infarction reperfusion. Eligible patients with STEMI reperfusion will be randomly divided into external cardiac shock therapy (ECSW) trial group and blank control group. The blank control group will receive optimal drug therapy, and the experimental group will receive optimal drug therapy combined with ECSW. The shock wave treatment plan will be 3-month therapy, specifically 1 week of treatment per month, 3 weeks of rest, 3 times of ECSW in each treatment week, respectively on the first day, the third day and the fifth day of the treatment week, lasting for 3 months and follow-up for 2 years. The primary endpoint will be to assess the 2-year improvement in all-cause death, re-hospitalization due to cardiovascular disease, major unintentional cerebrovascular events, including cardiogenic death, myocardial infarction, heart failure, arrhythmia, emergency coronary revascularization, and stroke in patients with STEMI reperfusion. Secondary endpoints will include improvements in angina pectoris, quality of life, cardiac structure and function, coronary microcirculation, and endothelial progenitor cell-derived miR-140-3p in relation to survival outcomes. TRIAL REGISTRATION NUMBER: ClinicalTrial.gov.org PRS:NCT05624203; Date of registration: November 12, 2022.

Golgi Phosphoprotein 3 Promotes Cervical Cancer Progression via Wnt/ß-catenin Mediated Epithelial-Mesenchymal Transition.

OBJECTIVE: Cervical cancer (CC) is one of the fatal malignancies affecting the life expectancy of women worldwide. Golgi Phosphoprotein 3 (GOLPH3) has been shown to play a key role in the development of various tumors. However, the role of GOLPH3 in the development of CC is unclear. METHODS: GOLPH3 levels were measured using quantitative real-time polymerase chain reaction (qRT-PCR) and western blot assays. Cell Counting Kit-8 (CCK-8) and colony formation assays were used to detect cell proliferation. Xenograft tumor models were used to explore the effects of GOLPH3 on tumor growth of mice, and immunohistochemistry assay was performed to determine the expression of GOLPH3 and Ki-67. Transwell assay was performed to evaluate cell migration and invasion. Western blot assay was used to analyze the signaling molecules-related proteins regulated by GOLPH3. RESULTS: GOLPH3 was upregulated in human CC tissues from the GEO database (GSE39001 and GSE63514), and further demonstrated that GOLPH3 level was elevated in human CC cells. GOLPH3 enhanced CC cell proliferation, and knockdown of GOLPH3 suppressed tumor growth and decreased Ki-67 level in xenograft mice. In addition, GOLPH3 aggravated the migration and invasion of CC cells. The data indicated that Wnt/ß-catenin signaling might be one of the key targets of GOLPH3. Blockage of the Wnt/ß-catenin pathway by XAV-939 significantly affected the effects of GOLPH3 on cell proliferation and epithelial-mesenchymal transition (EMT) related molecules, whereas LiCl (a Wnt/ß-catenin signal activator) reversed these above effects. CONCLUSION: GOLPH3 promotes cell proliferation, migration and invasion in CC, possibly by regulating the Wnt/ß-catenin signaling pathway, which may provide a new idea for the development of CC therapeutic targets.

[Prenatal diagnosis for a fetus with 5p deletion syndrome].

OBJECTIVE: To explore the genetic basis for a fetus with club foot detected upon mid-pregnancy ultrasonography. METHODS: Amniotic fluid of the fetus and peripheral blood samples of its parents were collected and subjected to G-banding karyotype analysis and copy number variation sequencing (CNV-seq). The result was verified by fluorescence in situ hybridization (FISH). RESULTS: The fetus and its parents all had a normal karyotype. CNV-seq analysis revealed that the fetus has harbored a 23.12 Mb on chromosome 5 and a 21.46 Mb duplication on chromosome 7. FISH assay has verified that its mother has carried a cryptic t(5;7)(p14.3;q33) translocation. CONCLUSION: CNV-seq combined with FISH can effectively detect cryptic chromosome aberrations, and can help to reduce severe birth defects and provide a basis for prenatal genetic counseling.

Circ_0042986 Presence Restrains Cervical Cancer Development via Upregulating PEG3 by Directly Targeting miR-582-3p.

The participation of circular RNAs (circRNAs) in carcinogenesis is widely established. Numerous circRNAs with aberrant expression in cervical cancer (CC) are identified by RNA sequencing, whereas the function of these circRNAs remains unclear. We thus aimed to unveil the effects and mechanisms of circ_0042986 in CC. Circ_0042986 with aberrant downregulation in CC was obtained from GSE10286 dataset. qPCR and western blotting were employed for the detection of circ_0042986, miR-582-3p, and paternally expressed 3 (PEG3) expressions. EdU assay, colony formation assay, wound healing assay, transwell assay, tube formation assay, and flow cytometry assay were applied for functional analyses. The potential binding site was ensured by dual-luciferase reporter analysis, and their binding relationship was verified by pull-down assay. The transplanted tumor models were constructed for in vivo function verification of circ_0042986. Our findings exposed that the downregulation of circ_0042986 was verified in clinical CC samples. Circ_0042986 overexpression largely attenuated CC cell growth, invasiveness, angiogenesis, and survival. MiR-582-3p was targeted by circ_0042986, and the inhibitory roles of circ_0042986 presence were partly abolished by miR-582-3p enrichment. MiR-582-3p combined with PEG3, and circ_0042986 increased PEG3 expression via decoying miR-582-3p. The interaction between miR-582-3p and PEG3 on CC cell functions was confirmed, evidenced by the reversal effects of PEG3 knockdown on miR-582-3p deficiency-inhibited cancer cell malignant behaviors. Circ_0042986 overexpression also limited tumorigenesis of CC in animal models. In summary, circ_0042986 overexpression decoyed miR-582-3p to increase PEG3 expression, thereby blocking the malignant progression of CC.

Dumbbell in the atrium: A rare case report of biatrial myxoma that crosses atrial septum diagnosed by multimodal echocardiography.

We reported a very rare case, a 59-year-old female whose heart myxoma was present in both atrium, the mass in biatrial was connected to each other at the oval foramen, resembling "dumbbell-like." By means of multimodality echocardiography techniques such as transthoracic echocardiography (TTE), contrast enhanced ultrasound (CEUS), and Real-time three-dimensional transesophageal echocardiography (RT-3D TEE), we have clarified the diagnosis. The patient underwent open-heart surgery to remove the biatrial myxoma which adhered to the oval fossa, with a slightly wider base and smooth lobulated surface. This case demonstrates the importance of multimodality echocardiography in the diagnosis of atypical myxomas.

Aggregation-Induced Emission Effect within Peroxyoxalate-Loaded Mesoporous Silica Nanoparticles for Efficient Harvest of Chemiluminescence Energy in Aqueous Solutions.

Aggregation-induced emission (AIE) molecules that can avoid the aggregation-caused quenching (ACQ) effect and break the concentration limit have been widely used for biosensing. Similar to fluorescence dyes, AIE molecules can be chemiexcited simply by a peroxyoxalate-based chemiluminescence (CL) reaction, but the hydrolysis of peroxyoxalate is often a problem in an aqueous solution. Herein, we report an AIE effect within peroxyoxalate-loaded silica nanoparticles (PMSNs) for an efficient harvest of CL energy as well as alleviation of bis(2,4,5-trichloro-6-carbopentoxyphenyl) oxalate (CPPO) hydrolysis. Peroxyoxalate (i.e., CPPO) and AIE molecules (i.e., 1,2-benzothiazol-2-triphenylamino acrylonitrile, BTPA) were loaded together within the mesoporous silica nanoparticles (MSNs) to synthesize the BTPA-PMSN nanocomposite. The BTPA-PMSNs not only allowed CPPO to be dispersed well in an aqueous solution but also avoided the hydrolysis of CPPO. Meanwhile, the proximity between BTPA and CPPO molecules in the mesopores of MSNs facilitated the BTPA aggregate to harvest the energy from CL intermediates. Hence, the CL system of BTPA-PMSNs can work efficiently in aqueous solutions at a physiological pH. The CL quantum yield of the BTPA-PMSN system was measured to be 9.91 × 10-5, about 20 000-fold higher than that obtained in the rhodamine B (RhB, a typical ACQ dye)-PMSN system. Using BTPA-PMSNs for H2O2 sensing, a limit of detection (LOD) as low as 5 nM can be achieved, 1000-fold lower than that achieved in the RhB-PMSNs system. Due to the feasibility of working at a physiological pH, this CL system is also quite suitable for the detection of oxidase substrates such as glucose and cholesterol. This BTPA-PMSN CL system with the merits of high CL quantum yield at a physiological pH is appealing for biosensing.

Cardiac Shock Wave Therapy Improves Ventricular Function by Relieving Fibrosis Through PI3K/Akt Signaling Pathway: Evidence From a Rat Model of Post-infarction Heart Failure.

Objection: Cumulative studies have identified the effectiveness of cardiac shock wave therapy (CSWT) in treating heart failure after acute myocardial infarction (AMI), but little have been discussed with regard to the beneficial effects of CSWT on anti-fibrosis along with the underlying mechanism. In this study, we investigated whether CSWT could reduce post-AMI fibrosis and further explored the molecular mechanism. Methods: Rat heart failure (HF) models induced by ligating the left anterior descending coronary artery were established and validated by echocardiography. Eligible animals were randomly categorized into five groups: the sham group, the HF group, the HF + CSWT group, the HF + LY294002 group, and the HF + CSWT + LY294002 group. The cardiac weight, serum level of BNP, NT-pro BNP and echocardiography parameters were measured to assess cardiac function in different groups. Masson's trichrome staining was used to assess the proportions of the fibrotic area. The expression level of CD34, αSMA was measured by RT-PCR, Immunohistochemistry and Immunofluorescent analyses and the level of PI3K/Akt was quantified by Immunohistochemistry and Western blotting. Results: The application of CSWT significantly improved cardiac function and reduced myocardial fibrosis and level of CD34 and αSMA, compared to the HF group. CSWT led to significant elevations of p-PI3K and p-Akt expression levels compared to that of the HF group and the inhibition of the PI3K/Akt pathway abolished the observed beneficial effects of CSWT. Conclusion: CSWT can facilitate the alleviation of cardiac fibrosis induced by AMI through the activation of PI3K/Akt signaling pathway.

Adsorption Promoted Aggregation-Induced Emission Showing Strong Dye Lateral Interactions.

Aggregation-induced emission (AIE) is a powerful method to produce fluorescence for a diverse range of applications. While most previous work induced aggregation by change of solvent, ionic strength, pH, or self-assembly, we herein explored adsorption-induced aggregation using 4,4'-(hydrazine-1,2-diylidene bis(methanylylidene)) bis(3-hydroxybenzoic acid) (HDBB) as an AIE luminogen. HDBB is known to aggregate with AIE at low pH but not at neutral pH, and its aggregation facilitates excited state intramolecular proton transfer for enhanced emission. Using a nonquenching nanomaterial, Y2O3 nanoparticles, HDBB showed sevenfold fluorescence increase at pH 7.0. Fluorescence lifetime showed that HDBB was in the aggregated state in the presence of Y2O3. For comparison, a fluorescent porphyrin compound showed that adsorption caused quenching after mixing with Y2O3, whereas other dyes such as fluorescein, calcein, and rhodamine B failed to be adsorbed by Y2O3. Adsorption did not follow a Langmuir isotherm, but it showed strong lateral HDBB interactions because adsorption was only achieved with a high concentration of HDBB. Adsorption was inhibited by salt and by phosphate, indicating the importance of electrostatic and metal-binding interactions. Comparisons were made with other nanomaterials, where graphene oxide and CeO2 quenched HDBB and a cationic liposome also enhanced its emission, although with a less red-shifted peak wavelength. This study provides a simple method to induce aggregation of an AIE dye and its aggregation in turn-enhanced adsorption.

Adsorption of cholesterol oxidase and entrapment of horseradish peroxidase in metal-organic frameworks for the colorimetric biosensing of cholesterol.

The co-immobilization of two enzymes onto single support commonly exhibits low efficiency due to the competition against limited sites. Water-stable metal-organic frameworks (MOFs) [i.e., PCN-333(Al)] with a high surface area and ultra-large cavities were employed to efficiently adsorb cholesterol oxidase (ChOx) and encapsulate horseradish peroxidase (HRP), respectively. The prepared PCN-333/ChOx&HRP was characterized through SEM, XRD, confocal microscopy, N2 adsorption isotherms, and thermal gravity analysis (TGA). The high surface area and high concentration of mesoporous cages resulted in the high loadings of both ChOx and HRP. The absorbed ChOx and the encapsulated HRP presented excellent activities without additional chemical modification. The immobilized enzymes were stable against protease digestion, organic solvents, temperature changes, and pH variation. Thus, a colorimetric biosensor for cholesterol detection was fabricated depending on cascade catalytic reactions of the immobilized bi-enzymes. An extended linear range from 0.0 to 40.0 µM with a low detection limit of 0.6 µM was obtained using the biosensor. The co-immobilization of the enzymes onto the surface and into the mesopores of MOFs provided a new and excellent platform for the development of highly stable and sensitive colorimetric biosensors.

[Construction of cell sheets of stem cells from apical papilla and characterization of their biological properties].

PURPOSE: Cell sheets of stem cells from apical papilla (SCAP) were constructed. Their histological and biological properties were studied to provide experimental basis for its application in dental pulp regeneration in immature permanent teeth. METHODS: SCAP were isolated and cultured from immature permanent third molars. Flow cytometry was used to evaluate surface marker expression of SCAP. Alizarin red S staining was used to test osteo/odontogenic differentiation capacity of SCAP. SCAP (at P3) were cultured in cell sheets medium for 14 days. Hematoxylin-eosin(H-E) staining was used for histological observation; flow cytometry analysis was used to test cell cycle of SCAP cell sheets; Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression levels of runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), and collagen I (Col-I). Western blot was used to detect the protein expression levels of Runx2 and ALP. SPSS 17.00 software package was used for statistical analysis. RESULTS: H-E staining showed that SCAP cell sheets contained multiple layers of cells (5 to 6 layers) with high cell density, retained tight junctions and secreted rich extracellular matrix. The cell cycle of SCAP cell sheets showed that G2+S phase was lower, while G1 phase was higher than SCAP, which indicated the proliferation rate of SCAP cell sheets was down-regulated (P＜0.05). Moreover, compared with SCAP, osteo/odontogenic differentiation capacity of SCAP cell sheets was significantly increased (P＜0.05). CONCLUSIONS: SCAP cell sheets secret rich extracellular matrix, and osteo/odontogenic differentiation capacity in SCAP cell sheets is higher than SCAP. SCAP cell sheets may possess more potentials in dental pulp regeneration.

Catalytic degradation of organophosphorous nerve agent simulants by polymer beads@graphene oxide with organophosphorus hydrolase-like activity based on rational design of functional bimetallic nuclear ligand.

The degradation of organophosphorous nerve agents is of primary concern due to the severe toxicity of these agents. Based on the active center of organophosphorus hydrolase (OPH), a bimetallic nuclear ligand, (5-vinyl-1,3-phenylene)bis(di(1H-imidazol-2-yl) methanol) (VPIM), was designed and synthesized, which contains four imidazole groups to mimic the four histidines at OPH active center. By grafting VPIM on graphene oxide (GO) surface via polymerization, the VPIM-polymer beads@GO was produced. The obtained OPH mimics has an impressive activity in dephosphorylation reactions (turnover frequency (TOF) towards paraoxon: 2.3 s-1). The synergistic catalytic effect of the bimetallic Zn2+ nuclear center and carboxyl groups on surface of GO possibly contributes to the high hydrolysis on organophosphate substrate. Thus, a biomimetic catalyst for efficient degradation of some organophosphorous nerve agent simulants, such as paraoxon and chlorpyrifos, was prepared by constructing catalytic active sites. The proposed mechanism and general synthetic strategy open new avenues for the engineering of functional GOs for biomimetic catalysts.

Thiolated DNA-templated silver nanoclusters with strong fluorescence emission and a long shelf-life.

Thiolated DNA (DNA-SH) was employed as a template in the synthesis and stabilization of AgNCs (DNA-SH-AgNCs). Resulting from the synergistic protective effect of specific Ag+-DNA interactions and Ag-S bonding, DNA-SH-AgNCs exhibited high quantum yields and resistance to oxidation.

Assembly of the active center of organophosphorus hydrolase in metal-organic frameworks via rational combination of functional ligands.

Different from popular mimics of bimetallic nuclear centers bridged by a hydroxide, a total coordination sphere of the active center of organophosphorus hydrolase was assembled in metal-organic frameworks by rational design and combination of ligands, which resulted in efficient destruction of nerve agent stimulants without a base as a co-catalyst.

Mimicking the Active Sites of Organophosphorus Hydrolase on the Backbone of Graphene Oxide to Destroy Nerve Agent Simulants.

Recent global military events, such as the conflict in Syria, have emphasized the need to find effective strategies to rapidly destroy organophosphorus-based nerve agents. In this work, we designed active site-engineered graphene oxide (GO) via polymerization (polymer bead-GOs) as organophosphorus hydrolase (OPH) mimetic hotspots for the rapid degradation of nerve agents. This hybrid catalyst has a high total turnover frequency value of 0.65 s-1 and good stability (94.8% activity maintained after 5 cycles). Mechanism investigations suggested that the high catalytic performance could be attributed to the synergistic effect among the clusters of imidazole and the presence of - COOH groups on the GO surface and Zn2+.

Association Analysis of Nonsyndromic Congenital Heart Disease and Tag Single Nucleotide Polymorphisms of TBX20 and Genes in the Ras-MAPK Pathway.

AIMS: The present study was performed to determine whether there are variants in TBX20 and genes of the Ras-MAPK pathway associated with nonsyndromic congenital heart disease (ns-CHD). MATERIALS AND METHODS: A total of 223 ns-CHD patients and 273 healthy controls from China were selected as study subjects to perform an association analysis using 22 tag single-nucleotide polymorphisms (tag SNPs) located either in one of three genes in the Ras-MAPK pathway (MAP2K2, BRAF, and RAF1) or the TBX20 gene that have previously been associated with syndromic congenital heart disease. RESULTS: The results showed that none of these tag SNPs are associated with ns-CHD. CONCLUSIONS: The results suggested that these disease-causing genes, which were previously discovered in familial cases, might not be the major genetic factors causing the development of ns-CHD in Chinese.

Label-free fluorometric detection of chymotrypsin activity using graphene oxide/nucleic-acid-stabilized silver nanoclusters hybrid materials.

Pancreatic function tests are used to determine the presence of chronic pancreatitis, particularly in the early stage of the disease. Chymotrypsin is an indicator of pancreatic function and is thus related to pancreatic diseases. A new fluorescent biosensing method for assay of chymotrypsin activity was developed using DNA (dC12)-templated silver nanoclusters and graphene oxide (GO). A peptide probe was also designed using chymotrypsin-cleavable amino acid sequence and a cysteine terminus. The peptide probe formed Ag-S bond to dC12-AgNCs to enhance the fluorescence of dC12-AgNCs. After the addition of GO, the peptide was adsorbed to the negative GO surface and the fluorescence of dC12-AgNCs was quenched by FRET. The peptide was then degraded into amino acid fragments upon addition of chymotrypsin; these fragments were released from the GO surface, and the FRET was terminated. The developed label-free method features lower cost and higher sensitivity to chymotrypsin activity assay compared with conventional fluorescence analysis. The method can be used to analyze chymotrypsin (as low as 3ng/mL, signal/noise =3) across a dynamic range of 0.0-50.0ng/mL. The proposed biosensing strategy can also be extended to other proteases by using different peptide substrates.

A peptide with a cysteine terminus: probe for label-free fluorescent detection of thrombin activity.

Thrombin has been implicated in atherosclerotic disease development. However, thrombin activity detection is currently limited because of the lack of convenient fluorescent probes. We developed a label-free fluorescent method to assay thrombin activity on the basis of a designed peptide probe with a thrombin-cleavable peptide sequence and a cysteine terminus. The peptide probe can be conjugated to DNA-templated silver nanoclusters (DNA-AgNCs) through Ag-S bonding; as a result, the fluorescence of DNA-AgNCs was enhanced. As the DNA-AgNCs-peptide conjugate was adsorbed to graphene oxide (GO), the enhanced fluorescence of DNA-AgNCs was quenched. Once the peptide probe was cleaved by thrombin, the resulting release of the DNA-AgNCs from the surface of GO restored the enhanced fluorescence. Thrombin can be determined with a linear range of 0.0-50.0 nM with a detection limit of 1 nM. The thrombin-sensitive probe with a cysteine terminus may be developed into probes to detect other proteases.