This paper was aimed to study the human plasma protein binding rate of diterpene ginkgolides meglumine injection.The equilibrium dialysis was used to determine the human plasma protein binding rate of ginkgolide A (GA),ginkgolide B (GB) and ginkgolide K (GK) in diterpene ginkgolides meglumine injection.The LC-MS/MS method was used for the content determination of ginkgolides.And then,the plasma protein binding rate was calculated.The results showed that there was no interference from other ingredients for the determination of ginkgolides.The calibration curve of the analytes was in good linearity in certain range of contents.The precision and stability of the analytes met the methodology requirements.After 8 h incubation,the human plasma protein binding rate of GA,GB and GK achieved balance.The human plasma protein binding rate of GA (0.34,1.70 and 8.51μg·mL-1) was 84.03%-88.11%; the human plasma protein binding rate of GB (0.62,3.09 and 15.5μg·mL-1) was 41.21%-53.56%; the human plasma protein binding rate of GK (0.04,0.20 and 1.01μg·mL-1) was 45.24%-59.59%.It was concluded that the method was simple,rapid and sensitive,which met the analysis requirement for biological samples.GA had a high plasma protein binding rate; GB and GK had medium plasma protein binding rate.

This study was aimed to investigate the induction effect ofRe-Du-Ning (RDN) Injection on rat liver microsome CYP450 enzymes. SD rats were randomly divided into the solvent control group, positive control group as well as the low, middle and high dose group of RDN (1, 2, 4 mL·kg-1·d-1). After drugs were administrated continuously for 7 days, the rats were sacrificed. The liver was weighed and prepared to microsomes. Meanwhile, the liver coefficients of rats were calculated. And the protein content was detected by BCA method. Finally, activities of five important subtypes of CYP450 enzymes such as CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A1/2 were measured by the“cocktail” method. The results showed that the levels of liver coefficients, microsome yield rate and activities of CYP450 subtypes increased significantly in the positive control group compared with the solvent control group (P < 0.01). There was no significant difference on the levels of liver coefficients, microsome yield and protein content between the low and middle dose group of RDN. However, there was significant difference on the levels of liver coefficients and microsome yield in the high dose group (P < 0.05). In terms of the influence on enzyme activity, RDN Injection can significantly induce the activities of CYP1A2 with dose dependence. It can induce the activities of CYP2C9 and CYP2C19 at the middle and high dose. However, there was no obvious influence on the activities of CYP3A1/2 and CYP2D6. It was concluded that the positive control group can obviously induce activities of CYP450, which can be used in the evaluation of induction experiments. RDN Injection had induction effect on CYP1A2, CYP2C9 and CYP2C19. But it had no influence on the activities of CYP3A1/2 and CYP2D6.

Objective To explore the clinical significance of epigenetic modification factors and metabolites in the early diagnosis and differential diagnosis of pancreatic cancer by detecting the levels of circulating nu-cleosomes,lactic acid and pyruvate in the serum of patients with pancreatic ductal adenocarcinoma,patients with other pancreatic tumors and healthy people.Methods A total of 26 patients with pancreatic ductal ade-nocarcinoma admitted to Peking University Third Hospital from January 2022 to April 2023 were selected as the experimental group.The other pancreatic tumor group included 8 cases of solid pseudopapillary tumor of the pancreas,11 cases of serous cystadenoma of the pancreas,and 9 cases of neuroendocrine tumor of the pan-creas.Another 26 healthy individuals who underwent outpatient examinations during the same period were se-lected as the health control group.Enzyme linked immunosorbent assay and targeted metabolomics were used to detect the expression levels of circulating nucleosomes,lactate,and pyruvate in serum,respectively.SPSS26.0 statistical software was used for multivariable statistical analysis to compare whether the difference in serum expression in the experimental group,other pancreatic tumor group and the health control group was statistically significant,and the metabonomics results of pancreatic cancer patients at different stages in the public database were analyzed by bioinformatics.Results There were statistically significant differences in the expression levels of lactate(P＜0.001),pyruvate(P＜0.001),and circulating nucleosomes(P＜0.001)be-tween the experimental group and the health control group,suggesting that the three had high sensitivity and specificity for the diagnosis of pancreatic ductal adenocarcinoma.Moreover,the expression levels of the three in the serum of the experimental group were significantly higher than those in the serum of other pancreatic tumor group,suggesting that these could be used to distinguish the types of pancreatic tumors.The serum ex-pression levels of the three in other pancreatic tumor group were significantly higher than those in the health control group,which were expected to become new risk indicators for pancreatic tumors.Among them,pyruvic acid showed the most significant difference in the combined model.Bioinformatics analysis indicated that pyru-vate and lactate were differential metabolites in patients with pancreatic cancer at different stages.Conclusion Circu-lating nucleosomes,lactic acid and pyruvate have high sensitivity and specificity among the three groups,which could be used as new markers of pancreatic cancer.

OBJECTIVE: To explore the genetic etiology of a patient with glycogen storage diseases. METHODS: Clinical data of child and his parents were collected. The genes associated with glycogen storage diseases were subjected to high-throughput sequencing to screen the variants. Candidate variant was validated by Sanger sequencing. Pathogenicity of the variant was predicted by bioinformatic analysis. RESULTS: High-throughput sequencing results showed that the boy has carried a hemizygous c.749C>T (p.S250L) variant of the PHKA2 gene. Sanger sequencing verified the results and confirmed that it was inherited from his mother. This variant was unreported previously and predicted to be pathogenic by bioinformatic analysis. CONCLUSION The patient was diagnosed with glycogen storage disease type IXa due to a novel c.749C>T (p.S250L) hemizygous variant of the PHKA2 gene. High-throughput sequencing can facilitate timely and accurate differential diagnosis of glycogen storage disease type IXa.
Child ; Family ; Genetic Testing ; Glycogen Storage Disease/pathology* ; High-Throughput Nucleotide Sequencing/methods* ; Humans ; Male ; Mutation ; Phosphorylase Kinase/genetics*