Short-term efficacy of empagliflozin in children with glycogen storage disease type Ⅰb / 中华儿科杂志

Objective: To analyze the short-time efficacy of empagliflozin in the treatment of glycogen storage disease type Ⅰb (GSD Ⅰb). Methods: In this prospective open-label single-arm study, the data of 4 patients were collected from the pediatric department in Peking Union Medical College Hospital from December 2020 to December 2022. All of them were diagnosed by gene sequencing and had neutropenia. These patients received empagliflozin treatment. Their clinical symptoms such as height and weight increase, abdominal pain, diarrhea, oral ulcer, infection times, and drug applications were recorded at 2 weeks, 1 month, 2 months, 3 months, 6 months, 9 months, 12 months, and 15 months after treatment to assess the therapeutic effect. The liquid chromatography-tandem mass spectrometry method was used to monitor the changes in 1, 5-anhydroglucitol (1, 5AG) concentration in plasma. At the same time, adverse reactions such as hypoglycemia and urinary tract infection were closely followed up and monitored. Results: The 4 patients with GSD Ⅰb were 15, 14, 4 and 14 years old, respectively at the beginning of empagliflozin treatment, and were followed up for 15, 15, 12 and 6 months, respectively. Maintenance dose range of empagliflozin was 0.24-0.39 mg/(kg·d). The frequency of diarrhea and abdominal pain decreased in cases 2, 3, and 4 at 1, 2 and 3 months of treatment, respectively. Their height and weight increased at different degrees.The absolute count of neutrophils increased from 0.84×109, 0.50×109, 0.48×109, 0.48×109/L to 1.48×109, 3.04×109, 1.10×109, 0.73×109/L, respectively. Granulocyte colony-stimulating factor was gradually reduced in 1 patients and stopped in 3 patient. Plasma 1, 5 AG levels in 2 children were significantly decreased after administration of empagliflozin (from 46.3 mg/L to 9.6 mg/L in case 2, and from 56.1 mg/L to 15.0 mg/L in case 3). All 4 patients had no adverse reactions such as hypoglycemia, abnormal liver or kidney function, or urinary system infection. Conclusion: In short-term observation, empagliflozin can improve the symptoms of GSD Ⅰb oral ulcers, abdominal pain, diarrhea, and recurrent infection, also can alleviate neutropenia and decrease 1, 5AG concentration in plasma, with favorable safety.

Application of micro-pen electrospray ionization tandem mass spectrometry in rapid detection of metabolic stability of testosterone and dextromethorphan / 药学学报

Probe electrospray ionization (PESI) is one of the typical types of ambient ionization technology, but its application in quantitative analysis is limited due to its poor sampling stability. Previously, we developed a new micro-pen electrospray ionization tandem mass spectrometry (μPen-ESI-MS/MS) method based on PESI. In this study, a μPen-ESI-MS/MS method to measure testosterone and dextromethorphan in liver microsome samples was developed and validated to further applicate in evaluating drug metabolism stability and CYP450 enzyme activity. A μPen-ESI-MS/MS method for detecting the CYP3A4 substrate testosterone and CYP2D6 substrate dextromethorphan in the liver microsome incubation system were developed, and the linearity, precision and accuracy of the method was validated. The validated method was further used to detect the metabolic stability of testosterone in the liver microsome incubation system. The results showed that the μPen-ESI-MS/MS had high efficiency with 0.3 min spraying time of each sample. The standard curve of the testosterone and dextromethorphan has good linearity (R2 > 0.99), the intra- and inter-batch accuracy of testosterone and dextromethorphan was 95.9%-109.3% and 90.5%-107.3%, respectively; the intra- and inter-batch precision was acceptable with RSD values of 2.4%-13.5% and 3.4%-12.1%. The half-lives of testosterone and dextromethorphan in the liver microsome incubation system were 12 min and 14 min, respectively. This study provided a rapid and sensitive μPen-ESI-MS/MS method for the assay of testosterone and dextromethorphan in liver microsome samples, and provided a new strategy for the evaluation of drug metabolism stability and CYP3A4/CYP2D6 activity.

Dynamic changes of cyclophosphamide-induced liver injury in mice / 药学学报

Cyclophosphamide (CPA) is one of the most commonly used alkylating agents in the treatment of malignant cancer. CPA is metabolized by cytochrome P450 enzymes into 4-hydroxycyclophosphamide in vivo which can exhibit anti-tumor activity. Metabolic activation of CPA can cause adverse reactions such as myelosuppression, cystitis, and liver injury. The aim of this study was to evaluate the dynamic changes of hepatic injury induced by CPA in mice. Male BALB/c mice were injected CPA (200 mg·kg-1) intraperitoneally. Both serum and liver samples were collected at 0, 2, 6, 12 and 24 hours after dosing. The animal experiment protocol was approved by the Institutional Animal Care and Use Committee at Sun Yat-sen University. The results showed that hepatotoxicity was observed at 2 hours after CPA dosing, and the most serious liver injury was measured at 12 hours. The level of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and malondialdehyde (MDA) was significantly increased, glutathione (GSH) level was significantly decreased, hepatocyte edema and vacuolar degeneration were widely observed in liver tissue, and began to recover 24 hours after dosing. In addition, due to oxidative stress damage caused by CPA, nuclear factor-erythroid 2-related factor 2 (NRF2) signaling pathway was activated and the mRNA and protein expression of its downstream targets such as quinine oxidoreductase 1 (NQO1), heme oxygenase-1 (HO-1), glutamate-cysteine ligase catalytic subunit (GCLC) and glutamate cysteine modifier subunit (GCLM) were up-regulated, which alleviated oxidative stress injury. In a summary, this study demonstrate the dynamic change of CPA-induced liver injury and the NRF2-mediated protective mechanisms, providing new insights into the CPA-induced liver injury.

Expression of Serum HMGB1 and CD14 mononuclear Toll-like Receptors in Children with Hemophagocytic Syndrome and Their Effect on Prognosis / 中国实验血液学杂志

OBJECTIVE@#To detect the expression of serum HMGB1 and CD14 monocyte Toll-like receptors in children with hemophagocytic syndrome (HPS), and to analyze its effect on the prognosis of children.@*METHODS@#Eight-three children with HPS admitted in Department of pediatrics of Wuhan Children's Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology were selected and enrolled in HPS group, at the same time 50 healthy children with same age were selected and enrolled in control group. The peripheral blood of children in 2 groups was collected. The flow cytometry was used to detect the expression of Toll-like receptors (TLR2, TLR4 and TLR6) in peripheral blood CD14 monocytes, the ELISA was used to detect the expression of HMGB1 in serum of peripheral blood. The relationship of TLR2, TLR4, TLR6 and HMGB1 expression with the clinical parameters, short-term therapentic efficacy and prognosis was analyzed, the relation of TLR2, TLR4 and TLR6 expression with HMGB1 also was analyzed.@*RESULTS@#The expression of TLR2, TLR4 and TLR6 on surface of CD14 monocytes and fluorescence intensity in HPS group were significantly higher than those in control group (P＜0.05). The serum HMGB1 level in HPS group was significantly higher than that in control group (P＜0.05). The expression levels of TLR2, TLR4, TLR6 and HMGB1 in HPS children who were in acute phase or had decrease of albumin or hemoglobin levels, thrombocytopenia, neutrophil absolute value to low and increase of triglycerides level, in HPS group all significantly increased, the difference in children with different sex and age was no statistically significant (P＞0.05). After treatment, the expressions of TLR2, TLR4, TLR6 and HMGB in CR and NAD groups was significantly lower than that before treatment (P＜0.05), while the expressions levels of TLR2, TLR4, TLR6 and HMGB in AD and RD groups were no statistically significant different from those before treatment (P＜0.05); the expressions levels of TLR2, TLR4, TLR6 and HMGB in dealth group all were higher than those in survival group (P＜0.05). The serum HMGB1 levels in HPS children positively correlated with expression of TLR2, TLR4 and TLR6 on CD14 monocytes (P＜ 0.05).@*CONCLUSION@#The expression rate and level of TLR2, TLR4 and TLR6 on CD14 monocytes in HPS children are significantly higher than those in healthy children.The expression levels of TLR2, TLR4, and TLR6 positively correlate with serum HMGB1 content, which provides reference for the diagnosis and prognosis of children with HPS.

Replication of association study between type 2 diabetes mellitus and IGF2BP2 in Han Chinese population / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The association between IGF2BP2 and type 2 diabetes mellitus (T2DM) has been repeatedly confirmed among different ethnic populations. However, in several genome-wide association studies (GWAS) from the Chinese Han population, the gene IGF2BP2 has not been replicated. The results of relevant studies for the association between IGF2BP2 and T2DM showed controversy in Chinese Han population. It is necessary to systematically evaluate the contribution of common variants in IGF2BP2 to T2DM in Chinese Han population.</p><p><b>METHODS</b>Two single-nucleotide polymorphisms (SNPs, rs4402960 and rs1470579) in IGF2BP2 were genotyped in Chinese Han population (3807 controls/4531 T2DM cases) by Illumina GoldenGate Indexing assay. The association between SNPs and T2DM was evaluated by multiple Logistic Regression analysis. A meta-analysis was used to estimate the effects of IGF2BP2 in 20854 Chinese Han individuals.</p><p><b>RESULTS</b>rs1470579 and rs4402960 were confirmed to have strong association with T2DM in the Chinese Han population (rs1470579 P = 1.80×10(-7), OR (95% CI) = 1.22 (1.14-1.32), rs4402960 P = 7.46×10(-9), OR (95% CI) = 1.26 (1.17-1.37), respectively). Moreover, 11 studies for rs4402960 were included in the meta-analysis and 7 studies for rs1470579. The meta-analysis also showed the association between T2DM and IGF2BP2 (rs1470579 OR of 1.15 (95% CI = 1.10-1.19), P < 0.0001 under an additive model and rs4402960 OR of 1.14 (95% CI = 1.10-1.18), P < 0.0001 under an additive model).</p><p><b>CONCLUSION</b>IGF2BP2 was strongly associated with the risk of T2DM in Chinese Han population.</p>