Method validation of Q-PCR detection of host residual DNA in antibody drug based on protein A magnetic beads.

The residual DNA derived from host cells in antibody drugs have potential safety risks. In this paper, the antibody in the test sample was removed by magnetic bead separation method, and the residual DNA were quantitatively determined by Q-PCR method. The residual DNA in the sample was analyzed according to the standard curve. We validated the species specificity, accuracy, precision, quantitative restrictions, reproducibility of this method. The results showed the linearrange was of 1 × 10-1～1 × 102 pg/µL and the curve linear was good, this method can specifically detect CHO cell DNA. Compared with the method of extracting residual DNA by magnetic beads, the method has the advantages of simplicity, rapidity and low cost, and can be used for quantitative determination of the residual host cell DNA in antibody drugs producted by CHO cells.

[Imaging measurement and clinical significance of the angle between the axis of pedicle and the plane of lamina in lower cervical vertebra].

OBJECTIVE: To explore the imaging measurement and clinical significance of the angle between the axis of pedicle and the plane of lamina in lower cervical vertebra.
 Methods: Three dimensional reconstruction of CT scan was performed in 30 patients with cervical deformity, and the angle between the axis of pedicle and the plane of lamina was measured with the specific reconstructed CT image of C3-C7.
 Results: 1) The left and right transverse angle of C3-C7 between the axis of pedicle and the ipsilateral plane of lamina were 98.3°±6.3°, 98.0°±5.1°, 97.5°±6.9°, 95.1°±5.0°, 85.8°±5.4° and 96.7°±8.2°, 98.7°±7.1°, 97.8°±3.6°, 93.2°±6.2°, 86.8°±5.7°, respectively, which showed a gradual decreasing trend. Meanwhile the angle of C3-C6 was more than 90 degrees and C7 was less than 90 degrees. In addition to C6 with C3 and C7 with other segments, the rest of the differences between the sections was not statistically significant (all P>0.05). 2) The left and right transverse angle of C3-C7 between the axis of pedicle and the pedicle of vertebral arch of lamina were 0.2°±4.5°, 1.2°±7.2°, -0.8°±6.8°, -3.3°±5.4°, -14.7°±4.0° and -1.6°±5.4°, 1.9°±4.6°, -0.5°±6.0°, -4.6°±5.3°, -13.7°±3.4°, respectively, which showed a first increasing and then reducing trend. Meanwhile the angle of C4 was maximum angle. In addition to C6 with C3, C6 with C4, and C7 with other segments, the differences between the sections was not statistically significant (all P>0.05). 3) The left and right sagittal angle of C3-C7 between the axis of pedicle and the ipsilateral plane of lamina were 77.7°±7.6°, 77.0°±7.1°, 85.3°±8.4°, 94.1°±2.2°, 94.9°±3.8° and 78.5°±7.1°, 76.2°±6.2°, 86.4°±6.4°, 94.0°±2.7°, 95.6°±3.8°, respectively, which showed a gradual increasing trend. The angle of C3-C4 was less than 90 degrees. C5 showed large variation and C6-C7 was more than 90 degrees. In addition to C3 with C4 and C6 with C7, the differences between the sections was statistically significant (all P<0.05). There was no significant difference between the two sides of the above indexes (all P>0.05).
 Conclusion: In low cervical vertebra, there is a certain angle relationship between the axis of pedicle and the plane of lamina, which can provide reference for the clinical determination of angle of pedicle screw insertion.

Electroacupuncture improves recovery after hemorrhagic brain injury by inducing the expression of angiopoietin-1 and -2 in rats.

BACKGROUND: Angiopoietin (Ang) is one of the major effectors of angiogenesis, playing a critical role in neurovascular remodeling after stroke. Acupuncture has been widely used for treating stroke in China for a long time. Recently, we have demonstrated that electroacupuncture (EA) can accelerate intracerebral hemorrhage (ICH)-induced angiogenesis in rats. In the present study, we investigated the effect of EA on the expression of Ang-1 and Ang-2 in the brain after ICH. METHODS: ICH was induced by stereotactic injection of collagenase type VII into the right globus pallidus. Adult male Sprague-Dawley rats were randomized into the following four groups: sham-operation (SHAM), stroke-no electroacupuncture (SNE), stroke-EA at the Zusanli acupoint (SEZ), and stroke-EA at a nonacupoint (SEN). EA was applied to the bilateral Zusanli (ST36) acupoint in the SEZ group and a nonacupoint in the SEN group. The expression of Ang-1 and Ang-2 was evaluated by immunohistochemistry and quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Some Ang-1 and Ang-2 immunoreactive microvessels with a dilated outline were detected in the perihematomal tissues after ICH, and the vessels extended into the clot from the surrounding area since day 7. The expression of Ang-1 increased notably as long as 2 weeks after ICH, while Ang-2 immunoreactivity declined at about 7 days following a striking upregulation at 3 days. EA at the Zusanli (ST36) acupoint upregulated the expression of Ang-1 and Ang-2 at both the protein and mRNA levels. However, EA at a nonacupoint had little effect on the expression of Ang-1 and Ang-2. CONCLUSIONS: Our data suggest that EA at the Zusanli (ST36) acupoint exerts neuroprotective effects on hemorrhagic stroke by upregulation of Ang-1 and Ang-2.

Comparison of the performance of two thermoluminescent dosimetry systems for the personal dose-equivalent Hp(10) measurement.

The performance of two thermoluminescent dosimetry systems (RGD-3D and RE2000) manufactured in China and Finland was compared. Both of these dosimetry systems demonstrated satisfactory results as their performance met the requirements of the standard. The two dosimetry systems showed similar performance in the energy response. The RGD-3D dosimetry system performed better in nonlinear response, minimum detectable level and blind sample tests, whereas the RE2000 dosimetry system showed better stability.

The dual role of LOXL4 in the pathogenesis and development of human malignant tumors: a narrative review.

Background and Objective: Lysyl oxidase-like protein 4 (LOXL4) is a secreted copper-dependent amine oxidase involved in the assembly and maintenance of extracellular matrix (ECM), playing a critical role in ECM formation and repair. Tumor-stroma interactions and ECM dysregulation are closely associated with the mechanisms underlying tumor initiation and progression. LOXL4 is the latest identified member of the lysyl oxidase (LOX) protein family. Currently, there is limited and controversial research on the role of LOXL4 in human malignancies. Its specific regulatory pathways, mechanisms, and roles in the occurrence, development, and treatment of malignancies remain incompletely understood. This article aims to illustrate the primary protein structure and the function of LOXL4 protein, and the relationship between LOXL4 protein and the occurrence and development of human malignant tumors to provide a reference for further clinical research. Methods: We searched the English literature on LOXL4 in the occurrence and development of various malignant tumors in PubMed and Web of Science. The search keywords include "cancer" "LOXL4" "malignant tumor" "tumorigenesis and development", etc. Key Content and Findings: LOXL4 is up-regulated in human gastric cancer, breast cancer, ovarian cancer, head and neck squamous cell carcinoma, esophageal carcinoma and colorectal cancer, but down-regulated in human bladder cancer and lung cancer and inhibits tumor growth. There are two conflicting reports of both upregulation and downregulation in hepatocellular carcinoma, suggesting that LOXL4 has a bidirectional effect of promoting or inhibiting cancer in different types of human malignant tumors. We further explore the application prospect of LOXL4 protein in the study of malignant tumors, laying a theoretical foundation for the clinical diagnosis, treatment and screening of prognostic markers of malignant tumors. Conclusions: LOXL4 exerts a bidirectional regulatory role, either inhibiting or promoting tumors depending on the type of cancer. We still need more research to further confirm the molecular mechanism of LOXL4 in cancer progression.

Time-course swRNA-seq uncovers a hierarchical gene regulatory network in controlling the response-repair-remodeling after wounding.

Wounding initiates intricate responses crucial for tissue repair and regeneration. Yet, the gene regulatory networks governing wound healing remain poorly understood. Here, employing single-worm RNA sequencing (swRNA-seq) across 12 time-points, we delineated a three-stage wound repair process in C. elegans: response, repair, and remodeling. Integrating diverse datasets, we constructed a dynamic regulatory network comprising 241 transcription regulators and their inferred targets. We identified potentially seven autoregulatory TFs and five cross-autoregulatory loops involving pqm-1 and jun-1. We revealed that TFs might interact with chromatin factors and form TF-TF combinatory modules via intrinsically disordered regions to enhance response robustness. We experimentally validated six regulators functioning in transcriptional and translocation-dependent manners. Notably, nhr-76, daf-16, nhr-84, and oef-1 are potentially required for efficient repair, while elt-2 may act as an inhibitor. These findings elucidate transcriptional responses and hierarchical regulatory networks during C. elegans wound repair, shedding light on mechanisms underlying tissue repair and regeneration.

Supra-Blan2 t score as a multisystem-based risk score to predict poor 3-month outcome in acute ischemic stroke patients with intravenous thrombolysis.

AIM: To develop and validate a novel weighted score integrating multisystem laboratory and clinical variables to predict poor 3-month outcome (mRS score of 3-6) in acute ischemic stroke (AIS) patients with intravenous thrombolysis (IVT) therapy. METHODS: We retrospectively analyzed data from Trial of Revascularization Treatment for Acute Ischemic Stroke study. The Supra-Blan2 t score was derived using the data on age, the National Institutes of Health Stroke Scale score, history of atrial fibrillation, blood sugar level, neutrophil count, direct bilirubin level, platelet-lymphocyte ratio, and TnI level in the derivation cohort of 433 patients, and validated in a cohort of 525 patients. Furthermore, we compared the performance of the Supra-Blan2 t score with DRAGON, TURN, and SPAN-100 scores. RESULTS: The discrimination capacity in the derivation and validation cohorts was good for poor 3-month outcome (the area under the curve was 0.821 and 0.843, respectively). The cumulative incidence of poor 3-month outcome significantly increased across risk categories in the derivation (low-risk, 9.2%; medium-risk, 17.4%; and high-risk, 58.8%) and validation cohorts (12.7%, 36.5%, and 73.6%, respectively). The performance of the Supra-Blan2 t score was similar to or superior to DRAGON, TURN, and SPAN-100 scores. CONCLUSION: The Supra-Blan2 t score, based on easily available multisystem laboratory and clinical variables, reliably predicted poor 3-month functional outcome in AIS patients treated with IVT therapy featuring good calibration and discrimination.

XBP1S associates with RUNX2 and regulates chondrocyte hypertrophy.

BMP2 (bone morphogenetic protein 2) is known to activate unfolded protein response signaling molecules, including XBP1S and ATF6. However, the influence on XBP1S and ATF6 in BMP2-induced chondrocyte differentiation has not yet been elucidated. In this study, we demonstrate that BMP2 mediates mild endoplasmic reticulum stress-activated ATF6 and directly regulates XBP1S splicing in the course of chondrogenesis. XBP1S is differentially expressed during BMP2-stimulated chondrocyte differentiation and exhibits prominent expression in growth plate chondrocytes. This expression is probably due to the activation of the XBP1 gene by ATF6 and splicing by IRE1a. ATF6 directly binds to the 5'-flanking regulatory region of the XBP1 gene at its consensus binding elements. Overexpression of XBP1S accelerates chondrocyte hypertrophy, as revealed by enhanced expression of type II collagen, type X collagen, and RUNX2; however, knockdown of XBP1S via the RNAi approach abolishes hypertrophic chondrocyte differentiation. In addition, XBP1S associates with RUNX2 and enhances RUNX2-induced chondrocyte hypertrophy. Altered expression of XBP1S in chondrocyte hypertrophy was accompanied by altered levels of IHH (Indian hedgehog) and PTHrP (parathyroid hormone-related peptide). Collectively, XBP1S may be a novel regulator of hypertrophic chondrocyte differentiation by 1) acting as a cofactor of RUNX2 and 2) affecting IHH/PTHrP signaling.

[The effect on BiP regulation of IRE1a promoter transcription activity and protein expression].

Usually, secreted or transmembrane proteins complete their three-dimension folding within endoplasmic reticulum (ER). Under the conditions of nutrient depletion, cell differentiation, or other stress statuses, misfolded or unfolded proteins aggregate within ER, and consequently cause ER stress and Unfolded Protein Response (UPR). In response to ER stress, BiP (Binding immunoglobulin protein) dissociates with IRE1a (Inositol-requiring kinase 1) and binds to unfolded proteins as a molecular chaperone in helping maintain their correct structure. Co-related to BiP's dissociation, IRE1a oglimerizes and activated its endoribonuclease domain by transautophosphorylation. Activated IRE1a then, by cleaving mRNA of Xbp1 and activating its transcription activity, triggers UPR. In this paper, in order to determine effect of BiP on transcription activity of IRE1a, we cloned promoter region of IRE1a into reporter gene analysis vector and found that BiP could upregulate promoter activity of IRE1a. Then, we constructed another 6 truncated promoter reporter vectors of IRE1a and pinpoint the core promoter activity region. Furthermore, both our RT-PCR and Western blot results showed that BiP could upregulate mRNA transcription level and protein expression level of IRE1a. Base on these findings, we can propose that, in order to alleviate ER stress caused by the misfolded or malfolded proteins, BiP could upregulate expression of IRE1a by increase its promoter activity. This study may suggest a novel signal pathway on IRE1a regulation in ER stress.

Trend and distribution analysis of occupational radiation exposure among medical practices in Chongqing, China (2008-2020).

The personal dose levels of medical radiation workers in Chongqing from 2008 to 2020 were investigated and analysed. The results showed that a total of 68 379 people were monitored from 2008 to 2020. The number of radiation workers increased year by year, and the number of female radiation workers increased rapidly. The monitoring data were analysed by Mann-Whitney test, Mann-Kendall test and Bonferroni method. The annual mean effective dose from 2008 to 2020 showed a decreasing trend (P < 0.0001). At the 5% significance level, six occupational categories showed a significant decreasing trend (P < 0.0001).The average annual effective dose for medical radiation workers in Chongqing in 2020 was 0.4482 mSv and 94.73% of radiation workers received annual doses less than the public dose limit (1 mSv). Personal dose monitoring results for most radiation workers were low. The protection of radiation workers in interventional radiology and nuclear medicine should be concerned.

CNS-LAND score: predicting early neurological deterioration after intravenous thrombolysis based on systemic responses and injury.

Importance: Early neurological deterioration (END) is a critical complication in acute ischemic stroke (AIS) patients receiving intravenous thrombolysis (IVT), with a need for reliable prediction tools to guide clinical interventions. Objective: This study aimed to develop and validate a rating scale, utilizing clinical variables and multisystem laboratory evaluation, to predict END after IVT. Design setting and participants: The Clinical Trial of Revascularization Treatment for Acute Ischemic Stroke (TRAIS) cohort enrolled consecutive AIS patients from 14 stroke centers in China (Jan 2018 to Jun 2022). Outcomes: END defined as NIHSS score increase >4 points or death within 24 h of stroke onset. Results: 1,213 patients (751 in the derivation cohort, 462 in the validation cohort) were included. The CNS-LAND score, a 9-point scale comprising seven variables (CK-MB, NIHSS score, systolic blood pressure, LDH, ALT, neutrophil, and D-dimer), demonstrated excellent differentiation of END (derivation cohort C statistic: 0.862; 95% CI: 0.796-0.928) and successful external validation (validation cohort C statistic: 0.851; 95% CI: 0.814-0.882). Risk stratification showed END risks of 2.1% vs. 29.5% (derivation cohort) and 2.6% vs. 31.2% (validation cohort) for scores 0-3 and 4-9, respectively. Conclusion: CNS-LAND score is a reliable predictor of END risk in AIS patients receiving IVT.

XBP1S protects cells from ER stress-induced apoptosis through Erk1/2 signaling pathway involving CHOP.

The mammalian unfolded protein response (UPR) protects the cell against the stress of misfolded proteins in the endoplasmic reticulum (ER), and the transcription factor X-box binding protein 1 spliced (XBP1S), a regulator of the UPR, is known to be important for ER stress (ERS)-mediated apoptosis and cell growth, but the molecular mechanism underlying these processes remains unexplored. Here, we report that knockdown of XBP1S by an siRNA silencing approach increased the expression of ERS-associated molecules. The overexpression of XBP1S stimulated, whereas its knockdown inhibited, cell proliferation in chondrocytes and chondrosarcoma cells; in addition, overexpression of XBP1S inhibited, while its repression enhanced, ERS-mediated apoptosis in chondrocytes and chondrosarcoma cells. Furthermore, XBP1S-mediated inhibition of apoptosis in response to ERS is through the Erk1/2 signaling pathway and down-regulation CHOP transcription factor. CHOP is one of the key downstream molecules known to be involved in ERS-mediated apoptosis. Collectively, these findings reveal a novel critical role of XBP1S in ERS-mediated apoptosis and the molecular mechanisms involved.

ATF4 and IRE1α inhibit DNA repair protein DNA-dependent protein kinase 1 induced by heat shock.

With the increase of environment temperature, more and more attentions are payed to the effects of heat stress. Cells under heat shock either are adapted to the condition or are damaged and dead. In this paper, we found that heat shock induced endoplasmic reticulum (ER) stress. ATF4, PERK, and IRE1α were induced by heat shock of 45 °C in the transcriptional level. Under the stress of 45 °C, PERK was phosphorylated and XBP1s was detected. The result indicated that heat shock could induce the ER stress. We found that heat shock of 45 °C induced the dysregulation of HSP70 and DNA-PKcs, and downregulated the expression of PARP1 and XRCC1. Further results showed that after the knockdown of ATF4 or IRE1α, the expression of DNA-PKcs and XRCC1 were increased. It was indicated that ATF4 and IRE1α could inhibit the expression of DNA-PKcs and XRCC1 under the heat stress. Our results suggested that heat shock could activate ER stress. IRE1α and ATF4, as the important ER stress molecules, could inhibit the expression of DNA repair proteins DNA-PKcs, XRCC1, and HSP70 under heat shock. Downregulation of DNA repair proteins could aggravate the cell damage that may cause cell apoptosis. This may explain that heat shock could increase the lethality of chemotherapeutic drugs on tumor cells.

Study on the effect of IRE1a on cell growth and apoptosis via modulation PLK1 in ER stress response.

The mammalian unfolded protein response (UPR) protects the cell against the stress of misfolded proteins in the endoplasmic reticulum (ER). Failure to adapt to ER stress causes the UPR to trigger apoptosis. Inositol-requiring enzyme-1a (IRE1a), as one of three unfolded protein sensors in UPR signaling pathways, senses ER unfolded proteins through an ER lumenal domain that becomes oligomerized during ER stress. It is known to be important for ER stress-mediated apoptosis and cell growth, but the exact molecular mechanism underlying these processes remains unexplored. In this study, we report that knockdown of IRE1a by an siRNA silencing approach enhanced, whereas its overexpression inhibited, cell proliferation in Hepatoma cells. Besides, overexpression of IRE1a induced, while its repression inhibited, ER stress-mediated apoptosis in Hepatomas cells. Furthermore, we found that overexpressed IRE1a can down-regulate Polo-like kinase 1(PLK1) from mRNA and protein two levels. IRE1a-mediated induction of apoptosis and inhibition of proliferation in response to ER stress is through downregulation PLK1, an early trigger for G2/M transition known to be participated in regulating cell proliferation and cell apoptosis. Collectively, these findings reveal a novel critical role of IRE1a in ER stress-mediated apoptosis and the molecular mechanisms involved. IRE1a may be a useful molecular target for the development of novel predictive and therapeutic strategies in cancer.

Expanding the phenotypic spectrum of KCNK4: From syndromic neurodevelopmental disorder to rolandic epilepsy.

The KCNK4 gene, predominantly distributed in neurons, plays an essential role in controlling the resting membrane potential and regulating cellular excitability. Previously, only two variants were identified to be associated with human disease, facial dysmorphism, hypertrichosis, epilepsy, intellectual/developmental delay, and gingival overgrowth (FHEIG) syndrome. In this study, we performed trio-based whole exon sequencing (WES) in a cohort of patients with epilepsy. Two de novo likely pathogenic variants were identified in two unrelated cases with heterogeneous phenotypes, including one with Rolandic epilepsy and one with the FHEIG syndrome. The two variants were predicted to be damaged by the majority of in silico algorithms. These variants showed no allele frequencies in controls and presented statistically higher frequencies in the case cohort than that in controls. The FHEIG syndrome-related variants were all located in the region with vital functions in stabilizing the conductive conformation, while the Rolandic epilepsy-related variant was distributed in the area with less impact on the conductive conformation. This study expanded the genetic and phenotypic spectrum of KCNK4. Phenotypic variations of KCNK4 are potentially associated with the molecular sub-regional effects. Carbamazepine/oxcarbazepine and valproate may be effective antiepileptic drugs for patients with KCNK4 variants.

Role and Mechanism of Gut Microbiota in Human Disease.

The human gut microbiome is a huge microbial community that plays an irreplaceable role in human life. With the further development of research, the influence of intestinal flora on human diseases has been gradually excavated. Gut microbiota (GM) dysbiosis has adverse health effects on the human body that will lead to a variety of chronic diseases. The underlying mechanisms of GM on human diseases are incredibly complicated. This review focuses on the regulation and mechanism of GM on neurodegenerative diseases, cardiovascular diseases, metabolic diseases and gastrointestinal diseases, thus providing a potential target for the prevention and treatment of disease.

Extracellular Matrix-Related Hubs Genes Have Adverse Effects on Gastric Adenocarcinoma Prognosis Based on Bioinformatics Analysis.

Gastric adenocarcinoma (GAC) is the most frequent type of stomach cancer, characterized by high heterogeneity and phenotypic diversity. Although many novel strategies have been developed for treating GAC, recurrence and metastasis rates are still high. Therefore, it is necessary to screen new potential biomarkers correlated with prognosis and novel molecular targets. Gene expression profiles were obtained from the from NCBI Gene Expression Omnibus (GEO) database. We conduct an integrated analysis using the online Venny website to explore candidate hub genes between differentially expressed genes (DEGs) of two datasets. Gene ontology (GO) and Kyoto Encyclopedia 18 of Genes and Genomes (KEGG) pathway enrichment analysis found that extracellular matrix plays an important role in GAC. In addition, we applied protein-protein interaction (PPI) network analysis by using the Search Tool for the Retrieval of Interacting Genes (STRING) and visualized with Cytoscape software. Furthermore, we employed Cytoscape software to analyze the interactive relationship of candidate gene for further analysis. We found that ECM related proteins played an important role in GAC, and 15 hub genes were extracted from 123 DEGs genes. There were four hub genes (bgn, vcan, col1a1 and timp1) predicted to be associated with poor prognosis among the 15 hub genes.

Dynamic compensation control strategy of DC bus voltage based on residual generator.

Aiming at the problem of bus voltage control in DC microgrid, a dynamic compensation control strategy based on a residual generator is designed to complete the voltage compensation of DC-DC converter. Firstly, based on the DC microgrid system architecture, the bus voltage fluctuations are analyzed theoretically, and then the DC-DC converter state-space mathematical models of the DC microgrid system are established to obtain the input-output relationship of the control system. Based on the theory of double coprime decomposition and Youla parameterization stable controller, the proposed control architecture based on the residual generator is obtained, and the output value generated by the current disturbance is compensated in reverse by applying model matching theory. The voltage loop compensation controller Q(s) is obtained by the linear matrix inequality method (LMI), and the current loop compensation controller H(s) is designed according to the dynamic structure diagram of the DC-DC converter. Hardware-in-the-loop simulation (HILS) results show that the architecture can improve the dynamic performance of the DC-DC converter without changing the original system structure parameters, and suppress the DC bus voltage fluctuations caused by load switching, power fluctuations, and AC-side load imbalances, and enhance the robustness of the system.

Bipolar Aggregation-Induced Electrochemiluminescence of Thiophene-Fused Conjugated Microporous Polymers.

Herein, we synthesize the thiophene tetraphenylethene-based conjugated microporous polymer (ThT-CMP) using the tetraphenylethylene derivative [i.e., 1,1,2,2-tetrakis(4-bromophenyl)ethane (TPBE)] and 2,5-thiophenediboronic acid as the precursors. The aggregation of TPBE in the ThT-CMP can induce a strong dual-band bipolar electrochemiluminescence (AIECL) emission at 554 nm (anodic) and 559 nm (cathodic) with tri-n-propylamine (TPrA) and S2O82- as the coreactants, respectively. The anodic and cathodic ECL efficiencies are measured to be 11.49 and 3.82% with respect to the standard of the Ru(bpy)32+/TPrA system, respectively. We further develop a dipolar ECL sensor to sensitively detect rhodamine B (RhB) based on resonance energy transfer. This ECL sensor possesses a large dynamic range and high sensitivity. This research provides a new avenue of designing organic structures with the characteristic of bipolar AIECL for the development of luminescent devices.