Glycolysis and de novo fatty acid synthesis cooperatively regulate pathological vascular smooth muscle cell phenotypic switching and neointimal hyperplasia.

Switching of vascular smooth muscle cells (VSMCs) from a contractile phenotype to a dedifferentiated (proliferative) phenotype contributes to neointima formation, which has been demonstrated to possess a tumor-like nature. Dysregulated glucose and lipid metabolism is recognized as a hallmark of tumors but has not thoroughly been elucidated in neointima formation. Here, we investigated the cooperative role of glycolysis and fatty acid synthesis in vascular injury-induced VSMC dedifferentiation and neointima formation. We found that the expression of hypoxia-inducible factor-1α (HIF-1α) and its target 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB3), a critical glycolytic enzyme, were induced in the neointimal VSMCs of human stenotic carotid arteries and wire-injured mouse carotid arteries. HIF-1α overexpression led to elevated glycolysis and resulted in a decreased contractile phenotype while promoting VSMC proliferation and activation of the mechanistic target of rapamycin complex 1 (mTORC1). Conversely, silencing Pfkfb3 had the opposite effects. Mechanistic studies demonstrated that glycolysis generates acetyl coenzyme A to fuel de novo fatty acid synthesis and mTORC1 activation. Whole-transcriptome sequencing analysis confirmed the increased expression of PFKFB3 and fatty acid synthetase (FASN) in dedifferentiated VSMCs. More importantly, FASN upregulation was observed in neointimal VSMCs of human stenotic carotid arteries. Finally, interfering with PFKFB3 or FASN suppressed vascular injury-induced mTORC1 activation, VSMC dedifferentiation, and neointima formation. Together, this study demonstrated that PFKFB3-mediated glycolytic reprogramming and FASN-mediated lipid metabolic reprogramming are distinctive features of VSMC phenotypic switching and could be potential therapeutic targets for treating vascular diseases with neointima formation. © 2023 The Pathological Society of Great Britain and Ireland.

Glycolysis inhibition ameliorates brain injury after ischemic stroke by promoting the function of myeloid-derived suppressor cells.

Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of immature myeloid cells which are immunosuppressive and glycolytically inactive in inflammatory diseases. However, it is unknown whether MDSCs contribute to ischemic stroke and how glycolysis regulates MDSC function in such a context. Here, we showed that MDSCs arise in the blood of patients at early phase of stroke. Similar results were observed in temporary middle cerebral artery occlusion-induced cerebral ischemic mice. Pharmaceutical exhaustion of MDSCs aggravated, while adoptive transfer of MDSCs rescued the ischemic brain injury. However, the differentiation of MDSCs into immunopotent myeloid cells which coincides with increased glycolysis was observed in the context of ischemic stroke. Mechanistically, the glycolytic product lactate autonomously induces MDSC differentiation through activation of mTORC1, and paracrinely activates Th1 and Th17 cells. Moreover, gene knockout or inhibition of the glycolytic enzyme PFKFB3 increased endogenous MDSCs by blocking their differentiation, and improved ischemic brain injury. Collectively, these results revealed that glycolytic switch decreases the immunosuppressive and neuroprotective role of MDSCs in ischemic stroke and pharmacological targeting MDSCs via glycolysis inhibition constitutes a promising therapeutic strategy for ischemic stroke.

Endothelial nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome regulation in atherosclerosis.

AIMS: The activation of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome in endothelial cells (ECs) contributes to vascular inflammation in atherosclerosis. Considering the high glycolytic rate of ECs, we delineated whether and how glycolysis determines endothelial NLRP3 inflammasome activation in atherosclerosis. METHODS AND RESULTS: Our results demonstrated a significant up-regulation of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3), a key regulator of glycolysis, in human and mouse atherosclerotic endothelium, which positively correlated with NLRP3 levels. Atherosclerotic stimuli up-regulated endothelial PFKFB3 expression via sterol regulatory element-binding protein 2 (SREBP2) transactivation. EC-selective haplodeficiency of Pfkfb3 in Apoe-/- mice resulted in reduced endothelial NLRP3 inflammasome activation and attenuation of atherogenesis. Mechanistic investigations revealed that PFKFB3-driven glycolysis increased the NADH content and induced oligomerization of C-terminal binding protein 1 (CtBP1), an NADH-sensitive transcriptional co-repressor. The monomer form, but not the oligomer form, of CtBP1 was found to associate with the transcriptional repressor Forkhead box P1 (FOXP1) and acted as a transrepressor of inflammasome components, including NLRP3, caspase-1, and interleukin-1ß (IL-1ß). Interfering with NADH-induced CtBP1 oligomerization restored its binding to FOXP1 and inhibited the glycolysis-dependent up-regulation of NLRP3, Caspase-1, and IL-1ß. Additionally, EC-specific overexpression of NADH-insensitive CtBP1 alleviates atherosclerosis. CONCLUSION: Our findings highlight the existence of a glycolysis-dependent NADH/CtBP/FOXP1-transrepression pathway that regulates endothelial NLRP3 inflammasome activation in atherogenesis. This pathway represents a potential target for selective PFKFB3 inhibitors or strategies aimed at disrupting CtBP1 oligomerization to modulate atherosclerosis.

Simultaneous analysis of dopamine and homovanillic acid by high-performance liquid chromatography with wall-jet/thin-layer electrochemical detection.

Wall-jet/thin-layer amperometric electrochemical detection (ECD) coupled with high-performance liquid chromatography (HPLC) was used here for the simultaneous analysis of dopamine (DA) and homovanillic acid (HVA) at a glassy carbon electrode. Compared with the conventional thin-layer mode and wall-jet mode, the presented wall-jet/thin-layer ECD has the advantages of enhanced capture of electroactive DA and HVA on the working electrode to give enhanced responses and more convenient washing/refreshment of the working electrode surface. Under optimized conditions, the HPLC-ECD calibration curves show good linearity from 0.01 to 100 µM for DA and HVA, and the limits of detection (LODs) obtained were 1.1 nM for DA and 0.7 nM for HVA which are lower than those obtained with an UV-vis detector and a commercial electrochemical detector. The method was tested on human urine with satisfactory results. The balance of response-signal, signal-background and noise level for our HPLC-ECD system is also discussed. In addition, a demethylation electrooxidation mechanism for HVA is suggested through potentiostatic bulk electrolysis, electrospray ionization-mass spectrometry, fluorescent spectrophotometry and cyclic voltammetry studies.

High-performance sorbents from ionic liquid activated walnut shell carbon: an investigation of adsorption and regeneration.

In this paper, walnut shells were selected to make activated charcoal using ionic activators. Based on the physical/chemical activation process and the properties of activated carbon products, the Fourier Transform Infrared reflection and Brunauer-Emmett-Teller analysis methods were adopted to comparatively analyse activation principles and pore-structure parameters. Also ciprofloxacin adsorption was compared among various activated carbon. Then, an absence of microporous structure in both walnut shells and their carbonized derivatives was found. Moreover, the specific surface area of activated carbon, prepared via KOH wet activation within physical/chemical procedures, attains a noteworthy 1787.06 m2 g-1, underlining its commendable adsorption performance. The specific surface areas of five distinct activated carbons, processed via ionic activation, extend from 1302.01 to 2214.06 m2 g-1. Concurrently, the micropore volumes span from 0.47 to 0.93 cm3 g-1. Obviously, the adsorption proficiency of ion-activated carbon markedly exceeds that of carbons activated physically or chemically. Of all materials investigated in this paper, ion-activated carbon D consistently exhibits superior performance, maintaining a ciprofloxacin removal rate nearing 100% at 40 °C. Remarkably, the maximum regeneration frequency of ion-activated carbons can reach up to 10 cycles. In conclusion, these five ion-activated carbons, demonstrating superior pore-structure parameters and adsorptive capacities, outperform those prepared through physical/chemical methods. They emerge as promising contenders for new, high-performing adsorbents.

Glycolysis Promotes Angiotensin II-Induced Aortic Remodeling Through Regulating Endothelial-to-Mesenchymal Transition via the Corepressor C-Terminal Binding Protein 1.

BACKGROUND: Endothelial dysfunction plays a crucial role in aortic remodeling. Aerobic glycolysis and endothelial-to-mesenchymal transition (EndoMT) have, respectively, been suggested to contribute to endothelial dysfunction in many cardiovascular diseases. Here, we tested the hypothesis that glycolytic reprogramming is critical for EndoMT induction in aortic remodeling through an epigenetic mechanism mediated by a transcriptional corepressor CtBP1 (C-terminal binding protein 1), a sensor of glycolysis-derived NADH. METHODS: EndoMT program, aortic remodeling, and endothelial expression of the glycolytic activator PFKFB3 (6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase isoform 3) were evaluated in Ang (angiotensin) II-infused mice. Mice with endothelial-specific Pfkfb3 deficiency or CtBP1 inactivation, immunoprecipitation, chromatin immunoprecipitation, and luciferase reporter assay were employed to elucidate whether and how PFKFB3/CtBP1 epigenetically controls EndoMT. RESULTS: The EndoMT program and increased endothelial PFKFB3 expression were induced in remodeled thoracic aortas. In TGF-ß (transforming growth factor-ß)-treated human endothelial cells, activated SMAD2/3 (SMAD Family Member 2/3) transcriptionally upregulated PFKFB3 expression. In turn, the TGF-ß/SMAD signaling and EndoMT were compromised by silencing or inhibition of PFKFB3. Mechanistic studies revealed that PFKFB3-mediated glycolysis increased NADH content and activated the NADH-sensitive CtBP1. Through interaction with the transcription repressor E2F4 (E2F Transcription Factor 4), CtBP1 enhanced E2F4-mediated transcriptional repression of SMURF2 (SMAD ubiquitin regulatory factor 2), a negative regulator of TGF-ß/SMAD2 signaling. Additionally, EC-specific Pfkfb3 deficiency or CtBP1 inactivation in mice led to attenuated Ang II-induced aortic remodeling. CONCLUSIONS: Our results demonstrate a glycolysis-mediated positive feedback loop of the TGF-ß signaling to induce EndoMT and indicate that therapeutically targeting endothelial PFKFB3 or CtBP1 activity could provide a basis for treating EndoMT-linked aortic remodeling.

Gene-dosage effect of Pfkfb3 on monocyte/macrophage biology in atherosclerosis.

BACKGROUND AND PURPOSE: Macrophage-rich atherosclerotic arteries are highly active in glycolysis. PFKFB3, a key glycolytic enzyme, has emerged as a potential therapeutic target in atherosclerosis. Small-molecule inhibitors of PFKFB3, such as 3PO and PFK158, have demonstrated efficacy in hampering atherogenesis in preclinical models. However, genetic studies elucidating the role of Pfkfb3 in atherogenesis need to be conducted to validate pharmacological findings and to unveil potential pharmacological side effects. EXPERIMENTAL APPROACH: Apoe-/- mice with global heterozygous or myeloid cell-specific Pfkfb3 deficiency were fed a Western diet (WD), after which atherosclerosis development was determined. Monocyte subsets in atherosclerotic mice and patients were examined by flow cytometry. Monocyte infiltration was assayed by a Ly6Chi monocyte-specific latex labelling procedure. In situ efferocytosis was assessed on mouse aortic root sections. Additionally, metabolic status, macrophage motility, efferocytosis, and involved mechanisms were analysed in peritoneal macrophages. KEY RESULTS: Global heterozygous or myeloid cell-specific Pfkfb3 deficiency reduced atherogenesis in Apoe-/- mice. Mechanistic studies showed that PFKFB3 controlled the proliferation and infiltration of proinflammatory monocytes. Moreover, PFKFB3 expression was associated with inflammatory monocyte expansion in patients with atherosclerotic coronary artery disease. Surprisingly, homozygous loss of Pfkfb3 impaired macrophage efferocytosis and exacerbated atherosclerosis in Apoe-/- mice. Mechanistically, PFKFB3-driven glycolysis was shown to be essential for actin polymerization, thus aiding the efferocytotic function of macrophages. CONCLUSION AND IMPLICATIONS: Collectively, these findings suggest the existence of a double-edged sword effect of myeloid PFKFB3 on the pathogenesis of atherosclerosis and highlight the need for caution in developing anti-atherosclerotic strategies that target PFKFB3.

Reduced expression of netrin-1 is associated with fetal growth restriction.

The objective of this study was to investigate the expression of netrin-1 in placenta from patients with fetal growth restriction (FGR) and its effect on the viability and apoptosis of human placental microvascular endothelial cells. Thirty-three patients with FGR (including eighteen severe cases) and twenty-four normal late pregnant women were investigated. The expression of netrin-1 in placental tissues was detected by employing immunohistochemistry, real-time PCR, and Western blotting. Human placental microvascular endothelial cells were isolated and, after treatment with netrin-1, examined for their viability and apoptosis by using MTT assay and flow cytometry. We demonstrated that the netrin-1 was present in placenta. Netrin-1 was significantly reduced in pregnant women with FGR as compared with the controls. Furthermore, netrin-1 enhanced the viability of human placental microvascular endothelial cells and inhibited their apoptosis. Netrin-1 may regulate the development of placental vessels and plays a key role in the pathogenesis of FGR.

Tumor Therapy: Targeted Substances Metabolism Reprogramming Induces Tumor Ferroptosis / 生物化学与生物物理进展

There are huge differences between tumor cells and normal cells in material metabolism, and tumor cells mainly show increased anabolism, decreased catabolism, and imbalance in substance metabolism. These differences provide the necessary material basis for the growth and reproduction of tumor cells, and also provide important targets for the treatment of tumors. Ferroptosis is an iron-dependent form of cell death characterized by an imbalance of iron-dependent lipid peroxidation and lipid membrane antioxidant systems in cells, resulting in excessive accumulation of lipid peroxide, causing damage to lipid membrane structure and loss of function, and ultimately cell death. The regulation of ferroptosis involves a variety of metabolic pathways, including glucose metabolism, lipid metabolism, amino acid metabolism, nucleotide metabolism and iron metabolism. In order for tumor cells to grow rapidly, their metabolic needs are more vigorous than those of normal cells. Tumor cells are metabolically reprogrammed to meet their rapidly proliferating material and energy needs. Metabolic reprogramming is mainly manifested in glycolysis and enhancement of pentose phosphate pathway, enhanced glutamine metabolism, increased nucleic acid synthesis, and iron metabolism tends to retain more intracellular iron. Metabolic reprogramming is accompanied by the production of reactive oxygen species and the activation of the antioxidant system. The state of high oxidative stress makes tumor cells more susceptible to redox imbalances, causing intracellular lipid peroxidation, which ultimately leads to ferroptosis. Therefore, in-depth study of the molecular mechanism and metabolic basis of ferroptosis is conducive to the development of new therapies to induce ferroptosis in cancer treatment. Ferroptosis, as a regulated form of cell death, can induce ferroptosis in tumor cells by pharmacologically or genetically targeting the metabolism of substances in tumor cells, which has great potential value in tumor treatment. This article summarizes the effects of cellular metabolism on ferroptosis in order to find new targets for tumor treatment and provide new ideas for clinical treatment.

Research Progress of Cell Senescence in Preeclampsia / 华中科技大学学报(医学版)

A well-developed placenta is the prerequisite for fetal development in utero while the placental dysplasia is the pathological basis of multiple pathological pregnancy,such as preeclampsia.Cell senescence is an irreversible state of cell cycle arrest.There is physiological senescence of the placenta in the progress of normal pregnancy.A variety of adverse factors can ac-celerate the pathological senescence of the placenta,resulting in placental dysplasia and dysfunction,and then induce multiple placental diseases such as preeclampsia.Recent studies have found that a variety of natural or synthetic anti-aging drugs also show a satisfying effect in preeclampsia disease models,suggesting that targeted cell senescence may be a potential mechanism for the treatment of preeclampsia.In this article,the latest literature was reviewed in terms of research status of cell senescence in the pathogenesis of preeclampsia.

Development of a Transformation System for the Medicinal Fungus Sanghuangporus baumii and Acquisition of High-Value Strain

To further explore the molecular mechanism of triterpenoid biosynthesis and acquire highvalue strain of Sanghuangporus baumii , the Agrobacterium tumefaciens-mediated transformation (ATMT) system was studied. The key triterpenoid biosynthesis-associated gene isopentenyl diphosphate isomerase (IDI) was transformed into S. baumii by ATMT system. Then, the qRT-PCR technique was used to analyze gene transcript level, and the widely targeted metabolomics was used to investigate individual triterpenoid content. Total triterpenoid content and anti-oxidant activity were determined by spectrophotometer. In this study, we for the first time established an efficient ATMT system and transferred the IDI gene into S. baumii. Relative to the wild-type (WT) strain, the IDI-transformant (IT) strain showed significantly higher transcript levels of IDI and total triterpenoid content. We then investigated individual triterpenoids in S. baumii, which led to the identification of 10 distinct triterpenoids. The contents of individual triterpenoids produced by the IT2 strain were 1.76–10.03 times higher than those produced by the WT strain. The triterpenoid production showed a significant positive correlation with the IDI gene expression. Besides, IT2 strain showed better anti-oxidant activity. The findings provide valuable information about the biosynthetic pathway of triterpenoids and provide a strategy for cultivating high-value S. baumii strains.

Preliminary analysis of qualitative changes in suspended red blood cells under different low-pressure and hypoxic conditions / 中国输血杂志

【Objective】 To evaluate the quality of suspended red blood cells (SRBC) under low-pressure and hypoxic conditions, in order to lay a theoretical foundation for airline delivery, air drop and storage of SRBC on plateau. 【Methods】 The low-pressure and hypoxic conditions (0.026 MPa and 0.047 MPa) were simulated by a pressure control device. SRBC were divided into 3 groups (5 bags/group, 1.5 U/bag). Each group was stored in 0.026 MPa environment at 2℃-6℃ for 24 hours (named 0.026 MPa), in 0.047MPa environment at 2℃-6℃ for 7 days (named 0.047 MPa), and in a blood storage refrigerator at 4℃ (named the control), respectively. The storage cell characteristics were examined on day 2, 9, 14, 28 and 35. 【Results】 We found that HCT, MCV, K+, Na+, FHb, hemolytic ratio, 2, 3-DPG and rheological properties in group 0.026 MPa and 0.047 MPa were not significant different compared with that in control (P>0.05). The consumptions of Glu in group 0.026 MPa and 0.047 MPa were significantly higher (P<0.05), and a transient increase in LAC concentration of group 0.026 MPa and 0.047MPa were observed, compared to the control. 【Conclusion】 The conditions of low-pressure and hypoxia have no significant effect on the quality of suspended red blood cells.

Prognostic Value of Pre-treatment Albumin/Fibrinogen Ratio in Patients with Diffuse Large B-cell Lymphoma / 中国实验血液学杂志

OBJECTIVE@#To investigate the value of pre-treatment albumin/fibrinogen ratio (AFR) on the prognosis of patients with diffuse large B-cell lymphoma (DLBCL).@*METHODS@#The data of DLBCL patients in the Affiliated Hospital of North Sichuan Medical College from April 2014 to March 2021 were retrieved, and 111 newly diagnosed patients who completed at least 4 cycles of R-CHOP or R-CHOP-like chemotherapy with complete data were included in the study. The clinical, laboratory examination and follow-up data of the patients were collected, and the receiver operating characteristic curve (ROC) was drawn according to patients' AFR before treatment and the survival status at the end of the follow-up, which could be used to preliminarily evaluate the predictive value of AFR for disease progression and patients' survival outcome. Furthermore, the correlation of AFR with the clinical and laboratory characteristics, progression-free survival (PFS) and overall survival (OS) was analyzed, and finally, univariate and multivariate Cox proportional hazard regression models were used to analyze factors affecting PFS and OS of DLBCL patients.@*RESULTS@#The ROC curve indicated that AFR level had a moderate predictive value for PFS and OS in DLBCL patients, with the area under the curve (AUC) of 0.616 (P =0.039) and 0.666 (P =0.004), respectively, and the optimal cut-off values were both 9.06 for PFS and OS. Compared with high-AFR (≥9.06) group, the low-AFR (<9.06) group had a higher proportion of patients with Lugano III-IV stage ( P <0.001), elevated lactate dehydrogenase (P =0.007) and B symptoms (P =0.038). The interim analysis of response showed that the overall response rate (ORR) in the high-AFR group was 89.7%, which was significantly higher than 62.8% in the low-AFR group (P =0.001). With a median follow-up of 18.5 (3-77) months, the median PFS of the high-AFR group was not reached, which was significantly superior to 17 months of the low-AFR group (P =0.009). Similarly, the median OS of high-AFR group was not reached, either, which was significantly superior to 48 months of the low-AFR group (P < 0.001). In multivariate Cox regression analysis, AFR <9.06 was an independent risk factor both for PFS and OS (HR PFS=2.047, P =0.039; HR OS=4.854, P =0.001).@*CONCLUSION@#Pre-treatment AFR has a significant value for the prognosis evaluation in newly diagnosed DLBCL patients.

Progress on standardized nutrition therapy for head and neck cancer during the peri-radiotherapeutic period / 中华放射肿瘤学杂志

Malnutrition is a common complication of cancer patients, and solving nutrition problems is still one of the challenging tasks in clinical practice. The incidence of malnutrition in head and neck cancer patients during the peri-radiotherapeutic period is high, which is not only related to disease-mediated metabolic disorders, complications and psychological factors, but also associated with the toxic and side effects induced by radiotherapy. Malnutrition will reduce the tolerance, accuracy, and therapeutic effects of radiotherapy, which in turn lowers the quality of life and even adversely affects the prognosis of disease. Medical nutrition therapy can improve the nutritional status of the body, ensure smooth progress of radiotherapy, and improve the efficacy of comprehensive cancer treatment. It is necessary and urgent to deliver standardized nutrition therapy and management of head and neck cancer patients during the peri-radiotherapeutic period. Nutritional risk screening, nutritional assessment, and acute radiation injury assessment are required to develop an individualized nutrition treatment plan and make dynamic adjustment. In this article, relevant literature of nutrition therapy for head and neck radiotherapy at home and abroad was summarized, and the standardized nutrition therapy for head and neck cancer patients during the peri-radiotherapeutic period was reviewed.

Sleep initiation patterns and their influence on sleep quality in infants and young children / 中国当代儿科杂志

OBJECTIVES@#To investigate the current status of sleep initiation patterns, influencing factors for sleep initiation patterns, and the influence of sleep initiation patterns on sleep quality in infants and young children.@*METHODS@#A total of 521 infants and young children, aged 0-35 months, who underwent physical examination at the outpatient service of the Department of Growth and Development in Shenzhen Bao'an Women's and Children's Hospital Affiliated to Jinan University were enrolled as subjects. A self-designed questionnaire and Brief Infant Sleep Questionnaire were used to collect the information on family background, parenting behaviors, and sleep quality in the past one week. A multivariate logistic regression analysis was used to evaluate the influencing factors for sleep initiation patterns. A multiple linear regression analysis was used to evaluate the influence of sleep initiation patterns on the number of nighttime awakenings.@*RESULTS@#Among the 521 infants and young children, 258 (49.5%) were breastfed/bottle fed to initiate sleep, 62 (11.9%) were rocked/held to initiate sleep, 39 (7.5%) slept independently, and 162 (31.1%) initiated sleep accompanied by parents. The multivariate logistic regression analysis showed that the children with breastfeeding and a younger age were more likely to be breastfed/bottle fed to initiate sleep (P<0.05) and that the children with a younger age were also more likely to be rocked/held to initiate sleep (P<0.05). The multiple linear regression analysis showed that sleep initiation with breastfeeding/bottle feeding significantly increased the number of nighttime awakenings (P<0.05).@*CONCLUSIONS@#Most infants and young children initiate sleep by being breastfed/bottle fed, rocked/held, or accompanied. The sleep initiation pattern is associated with the age of children and whether they are still breastfeeding. Sleep initiation with breastfeeding/bottle feeding can increase the number of nighttime awakenings. io.

Analysis of clinical phenotype and genotype of Chinese children with disorders of sex development / 中华儿科杂志

Objective: To explore the heterogeneity and correlation of clinical phenotypes and genotypes in children with disorders of sex development (DSD). Methods: A retrospective study of 1 235 patients with clinically proposed DSD in 36 pediatric medical institutions across the country from January 2017 to May 2021. After capturing 277 DSD-related candidate genes, second-generation sequencing was performed to analyzed the heterogeneity and correlation combined with clinical phenotypes. Results: Among 1 235 children with clinically proposed DSD, 980 were males and 255 were females of social gender at the time of initial diagnosis with the age ranged from 1 day of age to 17.92 years. A total of 443 children with pathogenic variants were detected through molecular genetic studies, with a positive detection rate of 35.9%. The most common clinical phenotypes were micropenis (455 cases), hypospadias (321 cases), and cryptorchidism (172 cases) and common mutations detected were in SRD5A2 gene (80 cases), AR gene (53 cases) and CYP21A2 gene (44 cases). Among them, the SRD5A2 mutation is the most common in children with simple micropenis and simple hypospadias, while the AMH mutation is the most common in children with simple cryptorchidism. Conclusions: The SRD5A2 mutation is the most common genetic variant in Chinese children with DSD, and micropenis, cryptorchidism, and hypospadias are the most common clinical phenotypes. Molecular diagnosis can provide clues about the biological basis of DSD, and can also guide clinicians to perform specific clinical examinations. Target sequence capture probes and next-generation sequencing technology can provide effective and economical genetic diagnosis for children with DSD.

Catalpol stimulates VEGF production via the JAK2/STAT3 pathway to improve angiogenesis in rats' stroke model.

ETHNOBOTANICAL RELEVANCE: Catalpol is the main active component of the radix from Rehmannia glutinosa Libosch, which has pleiotropic protective effects in neurodegenerative diseases, ischemic stroke, metabolic disorders and others AIM: Catalpol has been shown to have neuroprotective, neurorepair, and angiogenesis effects following ischemic brain injury. However, its molecular mechanisms are still poorly understood. In previous studies, the JAK2/STAT3 signaling pathway was found to play a role in neuroprotection and angiogenesis. This study investigated the role of catalpol in stimulating angiogenesis via the JAK2/STAT3 pathway after permanent focal cerebral ischemia (pMCAO). METHODS: Rats were subjected to right middle cerebral artery occlusion through electrocoagulation and were treated with catalpol (5mg/kg), AG490 was also used to inhibit STAT3 phosphorylation (pSTAT3). RESULTS: Following stroke, Catalpol improved the neuroethology deficit, increased the cerebral blood flow (CBF) of infarcted brain and upregulated EPO and EPOR. AG490 suppressed the phosphorylation of signal transducer and activator of transcription 3 (STAT3), ultimately inhibited VEGF mRNA expression, which reduced VEGF protein expression and inhibited stroke-induced angiogenesis. However, Catalpol enhanced stroke-induced STAT3 activation and subsequently restored STAT3 activity through the recovery of STAT3 binding to VEGF. Moreover, Catalpol reversed the effect of AG490 on STAT3 activation and nuclear translocation, restored the transcriptional activity of the VEGF promoter by recruiting STAT3 to the VEGF promoter, improved VEGF mRNA and protein expression, increased angiogenesis, reduced the difference in CBF between the infarcted and intact brain and ameliorated the neuroethology behaviors after stroke. CONCLUSION: Catalpol affects neuroprotection and angiogenesis via the JAK2/STAT3 signaling pathway, which is mediated by STAT3 activation and VEGF expression. Catalpol may be used as a potential therapeutic drug for stroke.

Second-generation Sequencing Analysis of Ph / 中国实验血液学杂志

OBJECTIVE@#To screen the core genes of Philadelphia chromosome positive/Ph like T-cell acute lymphoblastic leukemia (Ph@*METHODS@#The WES/RNA-seq examination results of Ph@*RESULTS@#For Ph@*CONCLUSION@#There are obviously abnormal DNA damage repair pathways in children with Ph

Predictive Value of Pre-treatment Serum Uric Acid Level for Prognosis in Newly Diagnosed Patients with Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To evaluate the predictive value of pre-treatment serum uric acid (sUA) level for the prognosis of newly diagnosed multiple myeloma (NDMM) patients.@*METHODS@#The NDMM patients admitted to our center from January 2014 to December 2018 were analyzed retrospectively, and 94 patients among them who were initially treated with bortezomib-based chemotherapy for at least 4 cycles were included in this study. Clinical characteristics, laboratory data and follow-up information were collected, and the predictive value of sUA on the overall survival (OS) of NDMM was evaluated by using receiver operating characteristic (ROC) curve based on the patient's pre-treatment sUA level and the survival status at the end of follow-up, and the correlation of the sUA level with patient's clinical, laboratory characteristics and overall survival (OS) was further analyzed. The univariate and multivariate Cox proportional-hazards model were used to identify the potential factors affecting OS.@*RESULTS@#ROC analysis showed that the area under the curve for predicting OS in NDMM patients with sUA level was 0.702 (P<0.001), and the optimal cut-off value was 455.4 μmol/L. Compared to patients with low sUA (<455.4 μmol/L), patients with higher sUA (≥455.4 μmol/L) were more likely to have international staging system (ISS) stage III disease, beta2-microglobulin (β@*CONCLUSION@#Pre-treatment sUA level is a potential biomarker for the prognosis evaluation in NDMM patients, which deserves a further exploration and verification.

Training the skills of obstetric forceps delivery for resident doctors by using simulation teaching method / 中华医学教育探索杂志

In this study, 15 second-year resident doctors who had standardized training of OBGYN were taken as research subjects to discuss the effect of training resident doctors with obstetrics forceps by using simulation teaching method. The simulation teaching process includes theoretical study, theoretical examination, simulation teaching of forceps skills and Assessment of forceps skills. In theoretical study section: before learning to use forceps, the students acquired the theoretical knowledge by flipped teaching micro-video mode. In theoretical examination section: the mastery of basic clinical knowledge of using forceps was evaluated through online assessment. In simulation teaching of forceps skills: students who scored 8/10 points were admitted to enter the simulation teaching process, and they had practical training of scenario simulation skills in Clinical Skills Center. In assessment of forceps skills: the mastery of forceps skills was evaluated by standardized forceps delivery procedure items. The results showed that the students in the simulation teaching group had excellent teaching assessment results, and the resident doctors had more confidence in operating forceps independently, and the teaching effect was ideal, which could further promote the simulation teaching of obstetric clinical skills.