Progress on the relationship of aldehyde dehydrogenase 2 with human diseases and its small-molecule activators / 药学实践杂志

Aldehyde dehydrogenase 2 (ALDH2) is one of important factors against from the damage under oxidative stress in human body. A high proportion of East Asians carry ALDH2 inactive mutation gene. There are many diseases closely related to ALDH2, such as cardiovascular diseases, neurodegenerative diseases and liver diseases. Recent studies also have found that ALDH2 is associated with ferroptosis. Therefore, ALDH2 has becoming a potential target for the treatment of the above related diseases. Several types of small molecule activators with potential value of clinical application have been reported. The research progress on the structure and function of ALDH2 , the relationship with human diseases and its activators were summarized in this paper.

Mechanism of catalpol regulating Th17 cell differentiation by interfering PKM2/LDHA expression / 中国药房

OBJECTIVE To investigate the mechanism of catalpol affecting the differentiation of helper T cell 17 （Th17） by interfering the expressions of pyruvate kinase M2 （PKM2） and lactate dehydrogenase A （LDHA）. METHODS The naive CD4+ T cells were selected from the spleen of C57BL/6 mice， and were differentiated into Th17 cells by adding directional differentiation stimulants for 72 hours. At the same time， the cells were treated with 0 （directed control）， 20， 40 and 80 μg/mL catalpol. The flow cytometry was used to detect the proportion of Th17 cell differentiation in cells； the colorimetric method was adopted to detect the levels of pyruvate and lactate in cell culture supernatant； mRNA expressions of retinoid-related orphan nuclear receptor gamma t （RORγt）， PKM2 and LDHA were detected by qRT-PCR method； Western blot was used to detect the expression levels of PKM2， LDHA， signal transducer and activator of transcription 3 （STAT3）， and phosphorylated STAT3 （p-STAT3） proteins in cells. RESULTS Compared with the directed control group， after 72 hours of treatment with 20， 40， 80 μg/mL catalpol， the differentiation ratio of Th17 cells were decreased by 6.74%， 8.41%， 9.24%， and the levels of pyruvate and lactate in the cell culture supernatant， the mRNA expressions of PKM2， LDHA and RORγt as well as the protein expressions of PKM2 and LDHA and the phosphorylation of STAT3 were significantly reduced （P＜0.05）. CONCLUSIONS Catalpol can reduce the glycolysis level by down-regulating the expressions of PKM2 and LDHA， thereby inhibiting the differentiation of Th17 cells.

Lactate dehydrogenase as a hematopoietic stem cell mobilization biomarker in autologous transplantation

ABSTRACT Introduction: Pre-apheresis peripheral blood CD34+ cell count (PBCD34+) is the most important predictor of good cell mobilization before hematopoietic stem cell transplantation, albeit flow cytometry is not always immediately available. Identification of surrogate markers can be useful. The CD34+ cells proliferate after mobilization, resulting in elevated lactate dehydrogenase (LDH) activity and correlating with the PBCD34+ count. Objective: To determine the LDH cut-off value at which adequate CD34+ cell mobilization is achieved and its diagnostic yield. Materials and methods: A total of 103 patients who received an autologous stem cell transplantation (ASCT) between January 2015 and January 2020 were included. Demographic and laboratory characteristics were obtained, including complete blood count, pre-apheresis PBCD34+ and LDH levels. Receiver operating characteristic (ROC) curves were performed to identify the optimal serum LDH activity cut-off points for ≥ 2 and ≥ 4 × 106 cells/kg post-mobilization CD34+ count and their diagnostic yield. Results: A post-mobilization serum LDH cut-off value of 462 U/L yielded a sensitivity (Se) = 86.8% (positive predictive value [PPV] = 72.7%), a pre- and post-mobilization serum LDH difference cut-off value of 387 U/L, an Se = 45.7% (PPV = 97%) and an LDH ratio of 2.46, with an Se = 47.1% (PPV = 97%) for an optimal mobilization count (CD34+ ≥ 4 × 106). Conclusion: The LDH measurement represents a fast and affordable way to predict PBCD34+ mobilization in cases where flow cytometry is not immediately available. According to the LDH diagnostic yield, it could be used as a surrogate marker in transplant centers, supporting the CD34+ count, which remains the gold standard.

CORRELATION OF SERUM FERRITIN AND LDH ON CRITICALLY ILL PATIENTS OF COVID-19 DISEASE DURING 1ST AND 2ND WAVE OF COVID 19 PANDEMIC IN TERTIARY CARE HOSPITAL, INDIA

In both the earlier waves of COVID-19 variants, severe and fatal respiratory disease like acute respiratory distress syndrome (ARDS) became more fatal in population with comorbid conditions. Therefore, early identi?cation of severe COVID-19 is very important for individual's precise management, including antiviral, oxygen support and intensive care unit (ICU) management. First case of COVID-19 got reported in the medical record of India on 30th January 2020 in a student who had returned from Wuhan, China. In 2020 and 2021 it was found that individuals with increased serum ferritin and LDH level landed up with severe and very severe COVID-19 if not treated timely and correctly. So correlation between S. Ferritin and LDH in 1st and 2nd wave was required to evaluate the condition of patients who remained admitted in critical care unit with or without comorbid conditions. This is hospital based cross- sectional observational study on 50-50 (total-100) critically ill patients admitted during 2020 and 2021 respectively. We found that In 2020 during the 1st wave serum LDH and serum Ferritin levels were signi?cantly high with the mean value of 481.65 U/L and 532.56 ng/ml respectively and in 2021 during 2nd wave serum LDH and serum Ferritin levels were again signi?cantly high with the mean value of 488.43 U/L and 667.27 ng/ml respectively. In 2020 patients with comorbid conditions showed S. LDH and Ferritin mean value of 543.47 U/L and 582.63 ng/ml respectively and in 2021 during 2nd wave it showed S.LDH and Ferritin levels mean value of 672.72 U/L and 727.38 ng/ml respectively. Both in?ammatory markers were signi?cantly more increased in the critically ill patients who presented with co-morbidities. This study will provide improved con?dence to health workers working in remote areas and COVID-19 hospitals in predicting transfer of COVID-19 patients to tertiary care hospitals for critical care management at the earliest.

Ventajas de la secuenciación de próxima generación sobre la hibridación fluorescente in situ para detectar la codeleción 1p/19q en oligodendrogliomas / Advantages of next generation sequencing over fluorescent in situ hybridization to detect 1p/19q codeletion in oligodendroglioma / Vantagens do sequenciamento de próxima geração sobre a hibridização fluorescente in situ para detectar codeleção 1p/19q em oligodendrogliomas

El perfil molecular de los gliomas permite garantizar la precisión del diagnóstico, informar el pronóstico e identificar opciones de tratamiento. Esta revisión tiene como objetivo exponer que con la secuenciación de próxima generación (NSG) el diagnóstico de los pacientes con oligodendrogliomas puede ser más exacto. Además, con un dispositivo de diagnóstico in vitro, basado en la NSG (F1CDx), en el que se utilizan los bloques de parafina de gliomas para analizar hasta 395 genes relacionados con cáncer (incluido IDH 1 y 2), se puede también informar la pérdida de la totalidad del brazo corto del cromosoma 1 y del brazo largo del cromosoma 19 (codeleción 1p/19q), a diferencia de la hibridación fluorescente in situ (FISH) que detecta desde la más mínima deleción, lo cual los hace sensibles pero no específicos ya que el FISH es incapaz de distinguir entre la pérdida de la totalidad del brazo del cromosoma y una deleción focal. Esta distinción es importante ya que la sobrevida es inferior en tumores con deleción parcial en rela­ción con los oligodendrogliomas, que tienen por definición la pérdida total de ambos cromosomas. Se hace también alusión a otras plataformas genómicas como GlioSeq y GLIO-DNA panel, que pueden cumplir la misma función. En conclusión, la F1CDx puede determinar con precisión 1p/19q con una concordancia del 96.7% frente a FISH. Los casos en que el FISH dio positivo y no concordaban con F1CDx, era porque no se trataba de oligodendrogliomas. F1CDx también analiza todos los genes que permiten la aproximación más exacta al diagnóstico de oligodendroglioma.

Molecular profiling of gliomas helps ensure diagnostic accuracy, inform prognosis, and identify treatment options. This review aims to show that with next generation sequencing (NGS) the diagnosis of patients with oligodendrogliomas can be more accurate. In addition, with an in vitro diagnostic device, based on NSG (F1CDx), in which glioma paraffin blocks are used to analyze up to 395 cancer-related genes (including IDH 1 and 2), it is also possible to report the loss of the entire short arm of chromosome 1 and the long arm of chromosome 19 (1p/19q codeletion), unlike fluorescence in situ hybridization (FISH) that detects even the slightest deletion, making them sensitive but not specific, as FISH is unable to distinguish between the loss of the entire arm of the chromosome and a focal deletion. This distinction is important since survival is lower in tumors with partial deletion compared to oligodendrogliomas, which by definition have the total loss of both chromosomes. Reference is also made to other genomic platforms such as GlioSeq and GLIO-DNA panel, which can fulfill the same function. In conclusion, the F1CDx can accurately determine 1p/19q with a concordance of 96.7% against FISH. The cases in which the FISH was positive and did not agree with F1CDx, it was because they were not oligodendrogliomas. F1CDx also analyzes all the genes that allow the most accurate approach to the diagnosis of oligodendroglioma.

O perfil molecular de gliomas ajuda a garantir a precisão do diagnóstico, informar o prognóstico e identificar as opções de tratamento. Esta revisão tem como objetivo mostrar que com o sequenciamento de próxima geração (NSG) o diagnóstico de pacientes com oligodendrogliomas pode ser mais preciso. Além disso, com um dispositivo de diagnóstico in vitro baseado em NSG (F1CDx), no qual blocos de parafina de glioma são usados para analisar até 395 genes relacionados ao câncer (incluindo IDH 1 e 2), também é possível relatar a perda do todo o braço curto do cromossomo 1 e o braço longo do cromossomo 19 (codeleção 1p/19q), ao contrário da hibridização fluorescente in situ(FISH) que detecta desde a menor deleção, o que os torna sensíveis, mas não específicos, pois o FISH é incapaz de distinguir entre a perda de todo o braço do cromossomo e uma deleção focal. Essa distinção é importante, pois a sobrevida é menor nos tumores com deleção parcial em relação aos oligodendrogliomas, que por definição apresentam a perda total de ambos os cromossomos. Também é feita referência a outras plataformas genômicas, como GlioSeq e painel GLIO-DNA, que podem cumprir a mesma função. Em conclusão, o F1CDx pode determinar com precisão 1p/19q com uma concordância de 96,7% versus FISH. Os casos em que FISH foi positivo e não concordaram com F1CDx, foi porque não eram oligodendrogliomas. O F1CDx também analisa todos os genes que permitem a abordagem mais precisa para o diagnóstico de oligodendroglioma.

Creatine supplementation for post-exercise muscle damage / Suplementação com creatina para danos musculares pós-exercício / Suplementación con creatina para daños musculares después del ejercicio

ABSTRACT Introduction: Exercise-induced muscle damage (EIMD) can occur from recent or unusual physical activity, leading to a temporary reduction in muscle function. And increased pain. Several articles indicate the positive impacts of creatine on EIMD. Objective: Evaluate the impact of creatine on EIMD. Methods: Online searches were performed in Scopus, Embase, Medline and Google scholar until March 2022. Results: Thirteen studies met the inclusion criteria. To assess the quality of the studies, the Cochrane collaboration system was used for risk and bias analysis. Due to the high heterogeneity of interventions and studies designed, a meta-analysis was not performed. The current paper reveals that creatine intake is preferable to inactive recovery and only a rest period between several harmful and exhausting physical activities. Conclusion: Benefits were attenuated in EIMD markers that reduce muscle operation and muscle strength loss after exercise. Level of evidence II; Therapeutic studies - Manuscript review.

RESUMO Introdução: O dano muscular induzido pelo exercício (EIMD) pode acontecer por atividade física recente ou não habitual e leva a uma redução temporária da função muscular. e aumento da dor. Vários artigos indicam impactos positivos da creatina sobre a EIMD. Objetivo: Avaliar o impacto da creatina sobre a EIMD. Métodos: Foram feitas pesquisas eletrônicas em Scopus, Embase, Medline e Google scholar até março de 2022. Resultados: Treze estudos preencheram os critérios de inclusão. Para avaliar a qualidade dos estudos, o sistema de colaboração Cochrane foi utilizado na análise de risco e viés. Devido à alta heterogeneidade de intervenções e estudos desenhados, a meta-análise não foi realizada. As informações do documento atual revelam que a ingestão de creatina é preferível a uma recuperação inativa e apenas um período de repouso entre diversas atividades físicas prejudiciais e exaustivas. Conclusão: Os benefícios evidenciaram-se atenuados nos marcadores EIMD que reduzem a operação muscular e a perda de força muscular após os exercícios. Nível de evidência II; Estudos terapêuticos - Revisão de manuscritos.

RESUMEN Introducción: el daño muscular inducido por el ejercicio (EIMD) puede producirse por una actividad física reciente o inusual y provoca una reducción temporal de la función muscular y un aumento del dolor. Varios artículos indican impactos positivos de la creatina en la EIMD. Objetivo: Evaluar el impacto de la creatina en la EIMD. Métodos: Se realizaron búsquedas electrónicas en Scopus, Embase, Medline y Google scholar hasta marzo de 2022. Resultados: Trece estudios cumplieron los criterios de inclusión. Para evaluar la calidad de los estudios, se utilizó el sistema de colaboración Cochrane para el análisis de riesgos y sesgos. Debido a la gran heterogeneidad de las intervenciones y de los estudios diseñados, no se realizó un metanálisis. La información del presente documento revela que la ingesta de creatina es preferible a una recuperación inactiva y sólo un período de descanso entre varias actividades físicas perjudiciales y agotadoras. Conclusión: Los beneficios se mostraron atenuados en los marcadores EIMD que reducen el funcionamiento muscular y la pérdida de fuerza muscular después del ejercicio. Nivel de evidencia II; Estudios terapéuticos - Revisión de manuscritos.

Impact of the lactate dehydrogenase in association with the International Staging System prognostic score in multiple myeloma patients treated in real life

Abstract Introduction Multiple myeloma is characterized by proliferation of clonal plasma cells. The identification of prognostics factors to identify patient's risk is important. Among the studied factors, it was identified of relevant importance the lactic dehydrogenase. Objectives To evaluate the impact of the value of DHL in combination with the score ISS in the medium patients overall survival (OS). Methods It is a retrospective cohort with 252 patients with MM recently-diagnosed that attendance in the institution of the study. Results To evaluate the association between DHL and ISS, we found 6 new groups to be analyzed: ISS I and normal DHL with medium overall survival not reached, and with DHL loud with medium OS of 69,8 months, ISS II and normal DHL with medium overall survival of 78,8 months and with DHL loud with medium OS of 73,9 months, ISS III and normal DHL with medium overall survival of 46,7 months and with DHL loud with medium OS of 45,5 months. Conclusion Through the association of ISS I and normal DHL, ISS III and high DHL and others combinations, we build a new score with superior impact prognostic in our population treated in real life.

Expression of acetaldehyde dehydrogenase 2 in endometrial carcinoma and its correlation with clinical characteristics of patients / 中国医师杂志

Objective:To detect the expression of acetaldehyde dehydrogenase (ALDH) 2 and microRNA (miR)-135b-3p in endometrial cancer (EC) tissues, and to explore their correlation with clinical characteristics.Methods:94 endometrial cancer tissue specimens (EC group) and 60 normal endometrial specimens (control group) were selected from Northwest Women′s and Children′s Hospital from February 2019 to February 2022. Real time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression changes of ALDH2 mRNA and miR-135b-3p in endometrial tissue of two groups, and immunohistochemistry was used to detect the positive expression rate of ALDH2 protein. The relationship between the expression levels of ALDH2 and miR-135b-3p and the clinicopathological characteristics was analyzed.Results:The expression of ALDH2 mRNA in the EC group (0.67±0.15) was lower than that in the control group (1.05±0.12), and the expression level of miR-135b-3p (1.52±0.26) was higher than that in the control group (1.01±0.06). The positive expression rate of ALDH2 in cancer tissue of EC group was 30.85%(29/94), which was lower than 51.67%(31/60) in normal endometrial tissue of the control group ( P<0.05). The expression levels of ALDH2 mRNA and miR-135b-3p in EC patients were related to International Federation of Obstetrics and Gynecology (FIGO) stage, lymph node metastasis, differentiation and myometrium invasion (all P<0.05), but not to age, pathological type, menopause, HPV infection, menarche age, parity, tumor length and BMI (all P>0.05). Conclusions:In EC tissues, the expression of ALDH2 is down-regulated and the expression of miR-135b-3p is up-regulated, which may be involved in the occurrence and development of EC.

Clinical efficacy of ateplase combined with heparin in the treatment of acute moderate- and high-risk pulmonary thromboembolism and its effects on arterial blood gas analysis and myocardial enzyme level / 中国基层医药

Objective:To investigate the clinical efficacy of alteplase combined with heparin in the treatment of acute moderate- and high-risk pulmonary thromboembolism and its effects on arterial blood gas analysis and myocardial enzyme level.Methods:Seventy-eight patients with acute moderate- and high-risk pulmonary thromboembolism who received treatment in Dongyang People's Hospital from January 2015 to January 2022 were retrospectively included in this study. They were divided into observation ( n = 39) and control ( n = 39) groups according to different treatment methods. The control group was treated with heparin, while the observation group was treated with alteplase based on heparin. All patients were treated for 7 days. Clinical efficacy as well as arterial blood gas analysis, myocardial enzymes, pulmonary artery pressure, and tricuspid annular plane systolic excursion pre- and post-treatment were compared between the two groups. Results:The total response rate in the observation group was significantly higher than that in the control group (94.87% vs. 76.92%, χ2 = 5.18, P < 0.05). After treatment, the partial pressure of carbon dioxide in the observation group was significantly lower than that in the control group [(36.24 ± 5.12) mmHg vs. (44.25 ± 3.78) mmHg, 1 mmHg = 0.133 kPa, t = 7.86, P < 0.05]. After treatment, the partial pressure of oxygen in the observation group was significantly higher than that in the control group [(78.82 ± 5.1) mmHg vs. (71.23 ± 4.89) mmHg, t = 6.66, P < 0.05]. After treatment, lactate dehydrogenase, creatine kinase, and creatine kinase isoenzyme in the observation group were (107.42 ± 15.45) U/L, (37.21 ± 10.84) U/L, and (12.28 ± 3.54) U/L, respectively, which were significantly lower than (189.94 ± 21.20) U/L, (65.42 ± 6.57) U/L, and (19.29 ± 3.08) U/L in the control group ( t = 19.64, 13.89, 9.33, all P < 0.001). After treatment, the pulmonary arterial pressure in the observation group was significantly lower than that in the control group [(32.24 ± 3.86) mmHg vs. (37.79 ± 5.17) mmHg, t = 5.37, P < 0.001]. Tricuspid annular plane systolic excursion in the observation group was significantly higher than that in the control group [(14.07 ± 1.27) mm vs. (12.63 ± 1.16) mm, t = 5.22, P < 0.001]. Conclusion:Ateplase combined with heparin has an obvious effect on acute moderate- and high-risk pulmonary thromboembolism. It can improve arterial blood gas analysis and reduce myocardial enzyme levels.

Establishment and application of activity determining methods for different subtypes of xanthine oxidoreductase / 药学学报

Xanthine oxidoreductase (XOR), the key enzyme catalyzing purine to produce uric acid, including two subtypes, xanthine dehydrogenase (XDH) and xanthine oxidase (XO), respectively, in vivo. Usually, XDH and XO can transform to each other. In this study, based on the principle that the subtype XO or XDH uses different electron acceptors, the methods for the measuring the activities of bovine milk XOR (pure enzyme) and its subtypes were established. The optimal concentrations of substrate xanthine (50 μmol·L-1) and electron acceptor NAD+ (50 μmol·L-1), pH value (7.80) were investigated. The ranges of the XOR, XO, XDH activity which could be determined were 0.97-17.5 U·L-1, 1-9 U·L-1, and 66-1 191 mU·L-1, respectively. Furthermore, the methods for determining the activities of XOR and its subtypes in mouse liver were established. The preparation of liver samples, the optimal concentrations of xanthine (100 μmol·L-1) and NAD+ (100 μmol·L-1) were researched. And the activity ranges of XOR, XO and XDH in mouse liver which could be determined were 0.67-3.98, 0.19-1.08, and 0.52-3.55 U·gprot-1, respectively. With the methods above, the effects of classic XOR inhibitor allopurinal (Allo) on XOR, XO and XDH from both milk and mouse liver were determined. All animal experiments have been approved by the Animal Experimental Center, Institute of Materia Medica, Chinese Academy of Medical Science and Peking Union Medical College (00003346). This study established new methods for the determination of XOR and its subtypes activity in pure enzyme system and in mouse liver, respectively, which were accurate and convenient. It laid the experimental foundation for exploring the different pathophysiological effects of XOR in the body and developing new XOR inhibitors.

Application value of C14∶1/C12∶1 for very long chain acyl-CoA dehydrogenase deficiency in newborn screening / 中华检验医学杂志

Objective:To establish the cut-off value of tetradecenoyl carnitine (C14∶1)/dodecenoyl carnitine(C12∶1) based on non-derivatized tandem mass spectrometry (MS/MS), and to explore the application value of C14∶1/C12∶1 to screen newborns for very long chain acyl-CoA dehydrogenase deficiency (VLCADD), determining the best combination of indicators for screening VLCADD.Methods:This retrospective study included data from 17 newborns with VLCADD detected by MS/MS and confirmed by acyl-CoA dehydrogenase very long chain ( ACADVL) gene detection, and 423 507 newborns with normal MS/MS results. The data from these newborns were collected from January 2014 to December 2021 as the newborns received neonatal screening in Nanjing Neonatal Disease Screening Center and Suzhou Neonatal Disease Screening Center. All newborns were divided into 3 groups: all newborns group, full-term newborns group and normal-birth-weight newborns group, and the cut-off values of C14∶1/C12∶1 for VLCADD in these 3 groups were determined by their receiver operating characteristic (ROC) curves individually. With these results, a total of 5 interpretation schemes were composed using different indicators alone or jointly: scheme 1 being C14∶1/C12∶1, scheme 2 being C14∶1, scheme 3 being C14∶1+C14∶1/C2+C14∶1/C16, scheme 4 being C14∶1/C12∶1+C14∶1, and scheme 5 being C14∶1/C12∶1+C14∶1+C14∶1/C2+C14∶1/C16. The detection rate, false-positive rate and positive predictive value of each scheme were calculated, and their screening efficiencies were statistically compared by Chi-square tests. Results:The cut-off values of C14∶1/C12∶1 for VLCADD in the 3 newborn groups were all 2.80. The detection rates of VLCADD with all 5 interpretation schemes were 17/17. Scheme 1 had the highest false positive rate [26.15‰ (11 075/423 524)] and the lowest positive predictive value [0.15% (17/11 092)]. Scheme 4 (Scheme 5) had the lowest false positive rate [0.02‰ (10/423 524)] and the highest positive predictive value [62.96% (17/27)]. Comparing scheme 4 (Scheme 5) with scheme 1, scheme 2 and scheme 3, the differences of false positive rate (χ2=302.30,11 191.50,32.06) and positive predictive value (χ2=102.51,3 485.61,13.83) were statistically significant (all P<0.001). Conclusion:C14∶1/C12∶1 was an effective auxiliary interpretive indicator for VLCADD in newborn screening, and the combination of C14∶1/C12∶1+C14∶1 was tested to be the best indicator for VLCADD screening based on non-derivatized tandem mass spectrometry.

Prenatal diagnosis of pyruvate dehydrogenase E1-α deficiency: a case report / 中华围产医学杂志

This article reported a case of pyruvate dehydrogenase E1-α deficiency suggested by abnormal brain development during prenatal ultrasound imaging. Prenatal ultrasound revealed a mild enlargement of bilateral cerebral ventricles and the possibility of intracranial hemorrhage in the fetus at 25 +1 weeks of gestation. MRI showed the fetus with absent corpus callosum, enlarged bilateral cerebral ventricles and paraventricular cysts. After genetic counseling and careful consideration, the couple opted for pregnancy termination. To clarify the cause of the disease, whole-exome sequencing was performed on the fetal skin to detect possible variants, and which revealed a frameshift mutation c.924_930dup(p.R311Gfs*5) in exon 10 of the PDHA1 gene. Sanger sequencing confirmed the mutation was a de novo pathogenic variant, indicating that the fetus was affected by pyruvate dehydrogenase E1-α deficiency.

Analysis of the clinical, pathological and genetic features of patients with myopathy-type very long chain acyl-coenzyme A dehydrogenase deficiency / 中华神经科杂志

Objective:To investigate the clinical, pathological and genetic characteristics of myopathy-type very long chain acyl-coenzyme A dehydrogenase deficiency (VLCADD).Methods:The detailed clinical data, muscle biopsy pathology and molecular results of 4 patients with genetically confirmed myopathy-type VLCADD admitted to Henan Provincial People′s Hospital and Xuanwu Hospital, Capital Medical University from June 2014 to November 2019 were retrospectively analyzed.Results:All of the 4 patients were late-onset myopathy-type VLCADD. The onset age ranged from 13 to 16 years, with a mean age of 14.5 years. The age at diagnosis ranged from 21 to 54 years, with a mean age of 42.5 years. The main clinical manifestation was repeated rhabdomyolysis, including myalgia, weakness and dark urine. Obvious somnolence was observerd in 1 patient. Muscle biopsy pathology revealed mild lipid accumulation, without vacuoles. Six ACADVL variations were detected in the 4 patients, including c.1283G>A (p.R428H), c.1532G>A (p.R511Q), c.833_835delAGA (p.K278del), c.1843C>T (p.R615 *), c.1748C>T (p.S583L) and c.1391C>T (p.T464I),among which c.1391C>T (p.T464I) was a novel variation, predicted to be likely pathogenic. Other 5 variations were reported pathogenic variations. Conclusions:Myopathy-type VLCADD is characterized by paroxysmal rhabdomyolysis and can be associated with somnolence. There is no specificity in muscle pathology. There are ACADVL variations, among which c.1391C>T is a novel variation.

Role of succinate dehydrogenase in hypoxic postconditioning-induced reduction of hypoxia-reoxygenation injury in myocardial cells of rats and the relationship with mito-K ATP / 中华麻醉学杂志

Objective:To evaluate the role of succinate dehydrogenase (SDH) in hypoxic postconditioning (HPC)-induced reduction of hypoxia-reoxygenation (H/R) injury in myocardial cells of rats and the relationship with mitochondrial ATP-sensitive potassium channels (mito-K ATP). Methods:Myocardial cells isolated from adult male Sprague-Dawley rats were cultured for 48 h and then divided into 7 groups ( n=24 each) using a random number table method: blank control group (Nor group), H/R group, SDHA-siRNA adenovirus+ H/R group (siRNA+ H/R group), HPC group, SDHA-siRNA adenovirus+ HPC group (siRNA+ HPC group), 5-HD+ HPC group, and SDHA-siRNA adenovirus+ 5-HD+ HPC group (siRNA+ 5-HD+ HPC group). Nor group was continuously cultured for 195 min under normoxic conditions. The H/R injury model was prepared by exposing the cells to hypoxia for 45 min in 5% CO 2 + 1% O 2 + 94% N 2, followed by reoxygenation for 150 min. The HPC method involved three cycles of 5 min reoxygenation/5 min hypoxia at the end of 45 min ischemia before 120 min reoxygenation. The mito-K ATP blocker 5-HD administration method involved adding 5-HD at a final concentration of 100 μmol/L at 30 min of hypoxia. The myocardial cells in each siRNA group were successfully transfected with SDHA-siRNA adenovirus to silence SDHA expression. The cell viability, calcium ion level, SDH activity, ATP content, degree of mitochondrial permeability transition pore (mPTP) opening, and mitochondrial membrane potential (MMP) were measured at the end of reoxygenation. Results:Compared with Nor group, the cell viability, ATP content and MMP were significantly decreased, and the degree of mPTP opening, level of calcium ion and activity of SDH were increased in H/R group ( P<0.05). Compared with H/R group, the cell viability, ATP content and MMP were significantly increased, and the degree of mPTP opening, calcium ion level and SDH activity were decreased in siRNA+ H/R group and HPC group ( P<0.05). Compared with HPC group, the cell viability, ATP content and MMP were significantly decreased, and the degree of mPTP opening, calcium ion level and SDH activity were increased in 5-HD+ HPC group ( P<0.05), and the cell viability, ATP content and MMP were significantly increased, and the degree of mPTP opening, calcium ion level and SDH activity were decreased in siRNA+ HPC group ( P<0.05). Compared with siRNA+ HPC group, the cell viability, ATP content and MMP were significantly decreased, the opening degree of mPTP and calcium ion level were increased ( P<0.05), and no significant change was found in the SDH activity in siRNA+ 5-HD+ HPC group ( P>0.05). Compared with 5-HD+ HPC group, the SDH activity was significantly decreased, and no significant change was found in the other parameters in siRNA+ 5-HD+ HPC group ( P>0.05). Conclusions:HPC alleviates H/R injury probably by reducing SDH activity and opening mito-K ATP in myocardial cells of rats.

Succinate dehydrogenase deficiency renal cell carcinoma: a case report / 中华泌尿外科杂志

Succinate dehydrogenase (SDH) defective renal cell carcinoma (RCC) is a new subtype of renal carcinoma newly identified by WHO(2016). Until now, only a few samples and a few cases have been reported retrospectively. This article reported a young female patient who was found to have a small tumor in the left kidney by physical examination and underwent left partial nephrectomy. The postoperative pathological result was SDH-RCC. There was no recurrence and metastasis of the tumor 3 months after operation.

The effect of tumor-associated macrophage on proliferation of renal carcinoma cells and related mechanism / 中华泌尿外科杂志

Objective:To investigate the effect of tumor-associated macrophage(TAM) on proliferation of renal carcinoma cells and its related mechanism.Methods:The model of TAM was established by stimulating human monocytic leukemia cell line THP-1 with phorbol myristate acetate (PMA), bacterial endotoxin (LPS) and interferon-γ (IFN- γ). Then the TAM model was co-cultured with carcinoma cell lines ACHN and 786-O in vitro .The cytokines IL-6, TNF-α and IL-1β in TAM supernatant were detected by enzyme-linked immunosorbent assay (ELISA). MTT method was used to detect the proliferation of ACHN and 786-O cells treated with supernatant of TAM or TAM/Tocilizumab. Western blot was used to detect lactate dehydrogenase A (LDHA) expression of both renal cancer cells co-cultured with TAM or TAM/Tocilizumab. The ACHN and 786-O cells with LDHA-overexpression and LDHA-knockdown were cultured in TAM supernatant in vitro. The cell proliferation was detected by MTT and the relative proliferation rate was calculated.Results:THP-1 cells was differentiated into TAM through the treatment of 80 ng/ml PMA combined with 20 ng/ml LPS and 20 ng/ml IFN- γ.The expression rate of CD68, a cell surface marker on TAM, was (36.2 ±4.5)%. When TAM was co-cultured with ACHN cells, the results of ELISA showed that the secretion of IL-6 in the supernatant was significantly elevated compared with that in the supernatant when ACHN cells cultured alone [(138.0 ±12.4) pg/ml and (19.7±4.9) pg/ml], and the secretion of TNF- α [(122.5 ±14.2) pg/ml and (12.6 ±2.3) pg/ml] and IL-1 β [(89.2 ±6.4) pg/ml and (69.2 ±3.5) pg/ml] were also significantly increased. The secretion of IL-6 [(119.2 ±14.8) pg/ml and (17.1 ±3.3) pg/ml], TNF- α [(122.6 ±14.4) pg/ml and (45.7 ±7.2) pg/ml] and IL-1 β [(95.1 ±11.8) pg/ml and (88.2 ±12.7) pg/ml] in the supernatant were also significantly elevated when 786-O cells co-cultured with TAM compared with 786-O cells cultured alone. After treated with the supernatant of TAM for 72 hours, the relative proliferation rates of ACHN and 786-O cells [(128.6 ±21.4)% and (124.2 ±19.7)%] were significantly higher than that of the control group (100.0%). At the same time, the expression of LDHA in ACHN and 786-O cells increased significantly. After 72 hours of treatment with the supernatant of TAM combined with tocilizumab, the relative proliferation rates of ACHN and 786-O cells [(76.5±13.7)% and (74.8±12.5)%] were significantly lower than that of the control group(100.0%), and the expression of LDHA was also significantly decreased at the same time. The relative proliferation rates of ACHN and 786-O cells in LDHA overexpression group [(121.5 ±17.2)% and (122.7±21.6)%]were significantly higher than that in blank-vector-transfection group[(93.3±10.7)% and (89.8±11.2)%], while the relative proliferation rates in LDHA-knockdown group [(61.4±11.2)% and (58.0 ±10.6)% ]were significantly lower than that in blank-vector-transfection group.Conclusions:By secreting IL-6, TAM can up-regulate the expression of LDHA and promote the proliferation of renal cancer cells.

Inhibitory Effect of Sophora alopecuroides Total Alkaloids and Two Monomer Alkaloids on PC12 Cell Activity / 中国实验方剂学杂志

ObjectiveTo compare the effects of total alkaloids， matrine， and sophoridine extracted from Sophora alopecuroides on the activity of pheochromocytoma cells （PC12 cells）. MethodThe effect of S. alopecuroides total alkaloids， matrine， and sophoridine at concentrations of 2， 1， 0.5， 0.25， 0.125， and 0.062 5 g·L-1 on the proliferation of PC12 cells was evaluated using the 3-（4，5-dimethylthiazol-2-yl）-2，5-diphenyl tetrazolium bromide （MTT） assay. The lactate dehydrogenase （LDH） leakage rate was measured by enzyme-linked immunosorbent assay （ELISA）. Flow cytometry was used to assess cell apoptosis rate， cell cycle distribution， and intracellular Ca2+ levels. Real-time quantitative polymerase chain reaction （Real-time PCR） was performed to determine the mRNA transcription levels of B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X protein （Bax）. Protein expression levels of apoptosis-related proteins Caspase-3， Caspase-8， Bcl-2， and Bax were detected by Western blot. ResultCompared to the control group， S. alopecuroides total alkaloids， matrine， and sophoridine inhibited the proliferation of PC12 cells， increased LDH leakage rate， enhanced intracellular Ca2+ fluorescence intensity， and induced cell apoptosis in concentration-dependent manner （P<0.05， P<0.01）. Among them， S. alopecuroides total alkaloids had the strongest inhibitory effect on cell proliferation and induction of apoptosis in PC12 cells （P<0.01）. After treatment with S. alopecuroides total alkaloids， matrine， and sophoridine， the cell cycle progression of PC12 cells was arrested at G1/G0 in the S. alopecuroides total alkaloids group， and at G1/S in the matrine and sophoridine groups. The expression levels of Bax mRNA were significantly increased （P<0.05， P<0.01）， while the expression levels of Bcl-2 mRNA were significantly decreased （P<0.05， P<0.01）. All treatments significantly downregulated the expression of the anti-apoptotic protein Bcl-2 （P<0.05， P<0.01） and upregulated the expression of the pro-apoptotic proteins Bax， Caspase-3， and Caspase-8 （P<0.05， P<0.01）， with the most significant protein expression changes observed in the S. alopecuroides total alkaloids group. ConclusionAmong the S. alopecuroides total alkaloids， matrine， and sophoridine， S. alopecuroides total alkaloids exhibit the strongest inhibitory effect on the activity of PC12 cells， and its mechanism of action may be related to the inhibition of anti-apoptotic protein expression， upregulation of pro-apoptotic protein expression， and activation of the mitochondrial Caspase-8 apoptotic pathway.

Baicalin inhibits PDK1 to mediate glucose metabolism reprogramming and intervene rheumatoid arthritis synovial inflammation / 药学学报

This study started from the effect of baicalin (BC), the main active component of the labiaceae plant Scutellaria baicalensis, on collagen-induced arthritis (CIA) in rats, to explore the mechanism of glucose metabolism reprogramming in fibroblast like synoviocytes (FLSs), a key effector cell of synovial inflammation in rheumatoid arthritis (RA). First of all, CIA rats and tumor necrosis factor-α (TNF-α)-induced RASFs in vitro and in vivo models were established, the arthritis index (AI) score and histopathological changes of CIA rats after BC administration were observed, and the levels of inflammatory factors in serum and cell supernatant were quantified by ELISA, immunocytochemistry and Western blot were used to detect the expression of G-protein-coupled receptor 81 (GPR81) and pyruvate dehydrogenase kinase 1 (PDK1) proteins. In addition, the kit was used to measure the levels of key products and enzyme activities in glucose metabolism reprogramming. The results showed that BC (50, 100 and 200 mg·kg-1) could alleviate the symptoms of arthritis in CIA rats in a dose-dependent manner, inhibit synovial hyperplasia, alleviate the infiltration of inflammatory cells, down-regulate the levels of pro-inflammatory factors TNF-α and interleukin (IL)-1β, and up-regulate the levels of anti-inflammatory factor IL-10 in CIA rats. At the same time, the secretion levels of lactate, pyruvate, acetyl-CoA, citrate and the activity of lactate dehydrogenase B (LDH-B) were decreased, and the expressions of GRP81 and PDK1 were down-regulated, suggesting that BC mediated the reprogramming process of glucose metabolism. However, when GPR81 inhibitor 3-OBA inhibited lactate uptake, the activity of LDH-B was significantly increased, suggesting that BC inhibited the expression of PDK1, a key enzyme in the reprogramming metabolism from glycolysis to oxidative phosphorylation. All animal experiments in this study were conducted in accordance with the ethical standards of the Laboratory Animal Care Center of Anhui University of Chinese Medicine (approval number: AHUCM-rats-2021049). These studies revealed that baicalin mediated metabolic reprogramming of RASFs from glycolysis to oxidative phosphorylation by inhibiting PDK1 protein expression, and alleviated joint inflammation in CIA rats.

Research progress on T cell glycolytic metabolism in oral lichen planus / 口腔疾病防治

@#Oral lichen planus (OLP) is a chronic inflammatory disease of the oral mucosa. The pathogenesis of OLP is still unclear. Immune abnormalities mediated by T cells and related cytokines play a crucial role in the pathogenesis of OLP. In recent years, glycolytic metabolism-related transporters, enzymes and regulators, such as glucose transporter-1 (Glut1), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), lactate dehydrogenase A (LDHA), mammalian target of rapamycin (mTOR) and hypoxia inducible factor-1α (HIF-1a), have attracted an increasing amount of attention in OLP by regulating the proliferation and differentiation of T cells and the secretion of inflammatory factors. It has been shown that 2-deoxy-D-glucose (2-DG) or rapamycin (RAPA) inhibits the glycolytic metabolism of T cells and then inhibits OLP. This article reviews the research progress of glycolytic metabolism-related transporters, enzymes and regulatory factors in OLP in recent years.

Advances in research on mechanism of lactate dehydrogenase B in tumors / 中国药科大学学报

@#Increased glycolysis is a major feature of metabolic reprogramming in cancer.Glycolysis provides not only energy for cancer cells but also necessary precursors for biosynthesis, which is important for promoting tumor growth.Cancer cells meet their own needs by regulating glycolytic enzymes, which play an active role in promoting cancer survival, metastasis, and invasion.Lactate dehydrogenase (LDH), as a key enzyme in glycolysis, consists of two subunits: lactate dehydrogenase A (LDHA) and lactate dehydrogenase B (LDHB).LDHA is known to play a key role in aerobic glycolysis and has been extensively studied, whereas less has been done on LDHB.However, at present, more and more reports have revealed the important effects of LDHB on the progression of various cancers.A large number of studies have shown that LDHB is abnormally expressed in a variety of cancers, which is related to the malignant progression of tumors.The article reviews the research progress of LDHB in recent ten years, including its regulatory mechanism in tumor, its relationship with cancer development and its role as a biomarker in clinical diagnosis of cancer, which provides some insight for further investigation of the mechanism of LDHB in cancer research.