OBJECTIVE To optimize the simmering technology of Rheum palmatum from Menghe Medical School and compare the difference of chemical components before and after processing. METHODS Using appearance score， the contents of gallic acid， 5-hydroxymethylfurfural （5-HMF）， sennoside A+sennoside B， combined anthraquinone and free anthraquinone as indexes， analytic hierarchy process （AHP）-entropy weight method was used to calculate the comprehensive score of evaluation indicators； the orthogonal experiment was designed to optimize the processing technology of simmering R. palmatum with fire temperature， simmering time， paper layer number and paper wrapping time as factors； validation test was conducted. The changes in the contents of five anthraquinones （aloe-emodin， rhein， emodin， chrysophanol， physcion）， five anthraquinone glycosides （barbaloin， rheinoside， rhubarb glycoside， emodin glycoside， and emodin methyl ether glycoside）， two sennosides （sennoside A， sennoside B）， gallic acid and 5-HMF were compared between simmered R. palmatum prepared by optimized technology and R. palmatum. RESULTS The optimal processing conditions of R. palmatum was as follows: each 80 g R. palmatum was wrapped with a layer of wet paper for 0.5 h， simmered on high heat for 20 min and then simmered at 140 ℃， the total simmering time was 2.5 h. The average comprehensive score of 3 validation tests was 94.10 （RSD＜1.0%）. After simmering， the contents of five anthraquinones and two sennosides were decreased significantly， while those of 5 free anthraquinones and gallic acid were increased to different extents； a new component 5-HMF was formed. CONCLUSIONS This study successfully optimizes the simmering technology of R. palmatum. There is a significant difference in the chemical components before and after processing， which can explain that simmering technology slows down the relase of R. palmatum and beneficiate it.

Inflammation plays a pivotal role in the etiology and progression of various diseases. In traditional Chinese medicine, the whole plants of Thesium chinense Turcz. and its preparations (e.g. Bairui Granules) have been employed to manage inflammatory conditions. While flavonoids were previously considered the primary anti-inflammatory components, other potentially active constituents have been largely overlooked and not thoroughly investigated. This study presents a novel finding that the total alkaloids of T. chinense (BC-Alk) are potent active substances underlying the traditional and clinical applications of T. chinense and Bairui Granules as anti-inflammatory agents. UPLC-MS/MS analysis identified the composition of BC-Alk as quinolizidine alkaloids. The anti-inflammatory efficacy of BC-Alk was evaluated using a lipopolysaccharide (LPS)-induced lung inflammation model in mice. Results demonstrated that BC-Alk significantly mitigated LPS-induced lung inflammation, attenuated the overproduction of IL-1β and the overproduction of inflammatory factors (TNF-α), and ameliorated lung tissue hyperplasia in mice in vivo. Mechanistic studies in vitro revealed that BC-Alk upregulated the expression of Nrf2 and its downstream proteins NQO1 and glutamate-cystine ligase and modifier subunit (GCLM), inhibited NF-κB phosphorylation, and suppressed NLRP3 activation. Collectively, these findings indicate that BC-Alk exerts potent inhibitory effects against lung inflammation by modulating Nrf2, NF-κB, and NLRP3 pathways. This study provides new insights into the anti-inflammatory constituents of T. chinense and Bairui Granules.
Animals ; Lipopolysaccharides/adverse effects* ; Alkaloids/pharmacology* ; NF-kappa B/metabolism* ; NF-E2-Related Factor 2/metabolism* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Mice ; Signal Transduction/drug effects* ; Anti-Inflammatory Agents/pharmacology* ; Male ; Mice, Inbred C57BL ; Humans ; Drugs, Chinese Herbal/administration & dosage* ; Pneumonia/genetics*

OBJECTIVE: To explore the clinical and genetic characteristics of two children diagnosed with two rare genetic diseases simultaneously. METHODS: Two children with comorbidity of two genetic diseases due to dual genetic mutations diagnosed at the Third Affiliated Hospital of Zhengzhou University respectively in May 2022 and March 2023 were selected as the study subjects. Clinical and genetic data of the two children were retrospectively analyzed. This study has been approved by the Ethics Committee of the Third Affiliated Hospital of Zhengzhou University (Ethic No. 2021-062-01). RESULTS: Child 1 was a 2-year-and-4-month-old boy whose clinical manifestations included facial dysmorphism, developmental delay, short stature, microcephaly, cleft palate, cryptorchidism, hypospadias, recurrent infections and immunological abnormalities. Whole exome sequencing revealed that he had harbored a heterozygous c.6595delT (p.Y2199Ifs*65) variant of the KMT2D gene and a heterozygous c.1892G>A (p.R631Q) variant of the PIK3R1 gene. This has led to a dual genetic diagnosis of Kabuki syndrome and PI3Kδ-related immunodeficiency type 36. Child 2 was a 15-year-old girl whose clinical manifestations included epilepsy, Albright's hereditary osteodystrophy, long body trunk, short limbs, hypocalcemia, hyperphosphatemia and hyperparathyroidism. The child also had a family history of short stature. Whole exome sequencing revealed that she had harbored a heterozygous c.2T>C (p.Met1?) variant of the GNAS gene and deletion of exons 2 to 6 of the SHOX gene. The two variants have led to dual diagnose of pseudohypoparathyroidism and X-linked idiopathic short stature. CONCLUSION When the clinical phenotype of a genetic disease is complex and cannot be fully explained with a single genetic variant, multiple pathogenic variants should be considered, and this may lead to the diagnosis of co-morbid genetic diseases. To adopt or supplement corresponding genetic testing in time and re-analyze the genetic data may facilitate accurate diagnosis of co-morbid genetic diseases.
Child, Preschool ; Female ; Humans ; Male ; Class Ia Phosphatidylinositol 3-Kinase/genetics* ; Comorbidity ; Exome Sequencing ; Mutation ; Rare Diseases/genetics* ; Retrospective Studies ; Adolescent

OBJECTIVE: To analyze the clinical characteristics and genotypic changes of six children with RARS2 gene variants. METHODS: The clinical data of 6 children with RARS2 gene variants diagnosed at the Third Affiliated Hospital of Zhengzhou University from January 2017 to August 2024 were collected. Genetic variants were detected using trio-whole exome sequencing. Genomic DNA was extracted from samples and subjected to high-throughput sequencing. Variants were detected and analyzed using relevant databases and software. Pathogenic variants were validated by Sanger sequencing. The protein structure encoded by a previously unreported variant was predicted using a SWISS-MODEL online server. This study was approved by the Medical Ethics Committee of the Third Affiliated Hospital of Zhengzhou University (Ethics No.: 2024-373-01). RESULTS: Among the six children, four were males and two were females, with the most recent follow-up age ranging from 1-year-and-1-month to 7 years old. The age of onset was under 1 year in all cases. All six children exhibited seizures, including infantile spasms in three, spasms and tonic spasms in one, and focal seizures in two. One child became seizure-free for 4 ~ 5 years following Valproic acid combined with topiramate and adrenocorticotropic hormone (ACTH) pulse therapy, but subsequently experienced a relapse. Another child has remained seizure-free for nearly one year with oral sodium valproate, levetiracetam, and a "cocktail" therapy. Seizures were not controlled in the remaining four children. Pontocerebellar hypoplasia was observed on neuroimaging in two children. All six patients exhibited severe psychomotor retardation. A total of 10 RARS2 gene variants were identified, three of which were previously unreported. CONCLUSION The predominant clinical features of Pontocerebellar hypoplasia associated with RARS2 gene variants include infantile onset, severe psychomotor retardation or regression, drug-resistant epilepsy, and feeding difficulties. The characteristic neuroimaging finding is pontocerebellar hypoplasia. However, its appearance may vary widely with time. The majority of affected children have a poor prognosis.
Humans ; Male ; Female ; Child, Preschool ; Infant ; Child ; Olivopontocerebellar Atrophies/genetics* ; Arginine-tRNA Ligase/genetics* ; Mutation ; Cerebellar Diseases

Objective:To investigate the effects of the Y-box binding protein 1(YBX1)on the proliferation and apoptosis of granulosa cells by regulating silent information regulator 1(SIRT1),and explore the differential ex-pression of YBX1 and SIRT1 of premature ovarian insufficiency(POI)patients and health controls,as well as its clinical significance.Methods:We recruited patients with POI and patients with normal ovarian function during in vitro fertilization(IVF)/intracytoplasmic sperm injection(ICSI)assisted pregnancy in the Department of Reproduc-tive Medical Center of the Fourth Affiliated Hospital of Jiangsu University from June 2022 to July 2023.The granu-losa cells and serum were collected.Western-Blot(WB)to detect YBX1 protein expression.Real-time quantitative polymerase chain reaction(RT-qPCR)to detect YBX1 and SIRT1 mRNA levels,and the correlation between the abundance of YBX1 and SIRT1 in serum and follicle stimulating hormone(FSH),luteinizing hormone(LH),estra-diol(E2),anti-Mullerian Hormone(AMH),and antral follicle count(AFC)were analyzed.5-bromo-2-deoxyuracil(EdU)and cell counting Kit-8(CCK8)were applied to detect cell proliferation;TdT-mediated dUTP-biotin nick end labeling assay(TUNEL)to detect cell apoptosis;RT-qPCR to detect cell proliferating cell nuclear antigen(PC-NA),B cell lymphoma 2(Bcl2),Bcl2 associated X protein(BAX),and cysteine-aspartate protease 3(Caspase-3)mRNA expression;RNA Immunoprecipitation(RIP)experiment to detect the interaction between YBX1 and SIRT1.Results:The expression of YBX1 protein and SIRT1 mRNA in granulosa cells and serum of POI patients were significantly lower than that of the control group(P＜0.05).The expression of YBX1 and SIRT1 mRNA in serum were negatively correlated with FSH,LH(r＜0,P＜0.05),and positively correlated with E2,AMH and AFC(r＞0,P＜0.05).Upregulating YBX1 in granulosa cells increased SIRT1 protein expression,SIRT1 mRNA stabili-ty,EdU positive rate,cell viability,PCNA mRNA expression level,Bcl2/BAX ratio(P＜0.05),while decreased Caspase-3 mRNA expression level,TUNEL positive rate(P＜0.05).Conclusions:The expression levels of YBX1 and SIRT1 in POI patients were significantly reduced and correlated with clinical ovarian function indicators.YBX1 can bind to SIRT1 mRNA and enhance its stability,which may promote the proliferation of granulosa cells,and in-hibit apoptosis.These findings suggested that YBX1 and SIRT1 are expected to become new targets for POI diag-nosis and treatment.

Objective:To investigate the effects of the Y-box binding protein 1(YBX1)on the proliferation and apoptosis of granulosa cells by regulating silent information regulator 1(SIRT1),and explore the differential ex-pression of YBX1 and SIRT1 of premature ovarian insufficiency(POI)patients and health controls,as well as its clinical significance.Methods:We recruited patients with POI and patients with normal ovarian function during in vitro fertilization(IVF)/intracytoplasmic sperm injection(ICSI)assisted pregnancy in the Department of Reproduc-tive Medical Center of the Fourth Affiliated Hospital of Jiangsu University from June 2022 to July 2023.The granu-losa cells and serum were collected.Western-Blot(WB)to detect YBX1 protein expression.Real-time quantitative polymerase chain reaction(RT-qPCR)to detect YBX1 and SIRT1 mRNA levels,and the correlation between the abundance of YBX1 and SIRT1 in serum and follicle stimulating hormone(FSH),luteinizing hormone(LH),estra-diol(E2),anti-Mullerian Hormone(AMH),and antral follicle count(AFC)were analyzed.5-bromo-2-deoxyuracil(EdU)and cell counting Kit-8(CCK8)were applied to detect cell proliferation;TdT-mediated dUTP-biotin nick end labeling assay(TUNEL)to detect cell apoptosis;RT-qPCR to detect cell proliferating cell nuclear antigen(PC-NA),B cell lymphoma 2(Bcl2),Bcl2 associated X protein(BAX),and cysteine-aspartate protease 3(Caspase-3)mRNA expression;RNA Immunoprecipitation(RIP)experiment to detect the interaction between YBX1 and SIRT1.Results:The expression of YBX1 protein and SIRT1 mRNA in granulosa cells and serum of POI patients were significantly lower than that of the control group(P＜0.05).The expression of YBX1 and SIRT1 mRNA in serum were negatively correlated with FSH,LH(r＜0,P＜0.05),and positively correlated with E2,AMH and AFC(r＞0,P＜0.05).Upregulating YBX1 in granulosa cells increased SIRT1 protein expression,SIRT1 mRNA stabili-ty,EdU positive rate,cell viability,PCNA mRNA expression level,Bcl2/BAX ratio(P＜0.05),while decreased Caspase-3 mRNA expression level,TUNEL positive rate(P＜0.05).Conclusions:The expression levels of YBX1 and SIRT1 in POI patients were significantly reduced and correlated with clinical ovarian function indicators.YBX1 can bind to SIRT1 mRNA and enhance its stability,which may promote the proliferation of granulosa cells,and in-hibit apoptosis.These findings suggested that YBX1 and SIRT1 are expected to become new targets for POI diag-nosis and treatment.

OBJECTIVE To mine adverse drug event （ADE） signals of lacosamide， and to provide references for clinically safe drug use. METHODS ADE data for lacosamide reported to the United States FDA adverse event reporting system from January 1， 2009， to December 31， 2022， were collected. Data mining was conducted using the reporting odds ratio method and Bayesian confidence propagation neural network method. Classification statistics were performed using the system organ class （SOC） and preferred terms （PT） from ADE terminology set of Medical Dictionary for Regulatory Activities （Version 25.0）. RESULTS A total of 21 360 lacosamide ADE reports were received， identifying 203 ADE signals across 24 SOCs， with 19 signals not included in the drug’s instruction. The top five PTs ranked by occurrence frequency were medication overdose， technical errors during device use， product use issues， intentional product misuse， and therapy discontinuation. The top five PTs ranked by signal strength were changes in seizure presentation type， congenital hypoplasia of depressor anguli oris muscle， multidrug resistance， brain surgery， and vagus nerve stimulator implantation. ADEs not recorded in the drug instruction included congenital hypoplasia of depressor anguli oris muscle， multidrug resistance， mitochondrial DNA mutation， dissociative identity disorder， and congenital auricular anomaly. CONCLUSIONS For lacosamide-induced ADEs that occur frequently and are already listed in the drug’s instructions， such as bradycardia and atrioventricular block， the clinical application should be careful and attentive， adjusting the dosage timely according to the patient’s condition to avoid severe ADEs. Newly discovered suspect ADEs， such as congenital hypoplasia of depressor anguli oris muscle， mitochondrial DNA mutation， overmature infant， dissociative identity disorder， pigmenturia， behavioral disorders， and dissociative disorders， should be vigilantly recognized to ensure the safety of drug use.

Objective To investigate the expression of circRAF1 in primary ovarian insufficiency(POI)and explore its effect on cell proliferation and apoptosis of human ovarian granulosa cells(GCs)line(KGN cells).Methods The expression of circRAF1 in GCs and serum of patients with normal ovarian reserve function(n = 50)and patients with POI(n = 50)were detected with RT-qPCR.The correlation of circRAF1 with ovarian reserve function indexes was analyzed.Small interfering RNA(siRNA)targeting circRAF1 was constructed and trans-fected into KGN cells,with the cell proliferation detected by CCK-8 and EdU assay,and the cell apoptosis detected by JC-1 and Tunel assay.The mRNA and protein levels of genes related to cell proliferation and apoptosis(FSHR,PCNA,Bcl-2,Casp-9,Bax)were detected by RT-qPCR and WB.Results The expression of circRAF1 decreased in GCs and serum of POI patients.The expression of circRAF1 was positively correlated with serum E2 and AMH levels(P<0.001),but negatively correlated with serum FSH and LH levels(P<0.001).At the same time,the expression of circRAF1 was positively correlated with AFC(P<0.001).Interfering with the expression of circRAF1 could inhibit the proliferation of KGN cells and promote their apoptosis.Conclusion The expression of circRAF1 in the GCs and serum of POI patients is down-regulated,which is correlated with the decline of ovarian reserve function.Interfering with circRAF1 can inhibit the proliferation of GCs and promote their apoptosis.

Objective:To investigate the effects of the Y-box binding protein 1(YBX1)on the proliferation and apoptosis of granulosa cells by regulating silent information regulator 1(SIRT1),and explore the differential ex-pression of YBX1 and SIRT1 of premature ovarian insufficiency(POI)patients and health controls,as well as its clinical significance.Methods:We recruited patients with POI and patients with normal ovarian function during in vitro fertilization(IVF)/intracytoplasmic sperm injection(ICSI)assisted pregnancy in the Department of Reproduc-tive Medical Center of the Fourth Affiliated Hospital of Jiangsu University from June 2022 to July 2023.The granu-losa cells and serum were collected.Western-Blot(WB)to detect YBX1 protein expression.Real-time quantitative polymerase chain reaction(RT-qPCR)to detect YBX1 and SIRT1 mRNA levels,and the correlation between the abundance of YBX1 and SIRT1 in serum and follicle stimulating hormone(FSH),luteinizing hormone(LH),estra-diol(E2),anti-Mullerian Hormone(AMH),and antral follicle count(AFC)were analyzed.5-bromo-2-deoxyuracil(EdU)and cell counting Kit-8(CCK8)were applied to detect cell proliferation;TdT-mediated dUTP-biotin nick end labeling assay(TUNEL)to detect cell apoptosis;RT-qPCR to detect cell proliferating cell nuclear antigen(PC-NA),B cell lymphoma 2(Bcl2),Bcl2 associated X protein(BAX),and cysteine-aspartate protease 3(Caspase-3)mRNA expression;RNA Immunoprecipitation(RIP)experiment to detect the interaction between YBX1 and SIRT1.Results:The expression of YBX1 protein and SIRT1 mRNA in granulosa cells and serum of POI patients were significantly lower than that of the control group(P＜0.05).The expression of YBX1 and SIRT1 mRNA in serum were negatively correlated with FSH,LH(r＜0,P＜0.05),and positively correlated with E2,AMH and AFC(r＞0,P＜0.05).Upregulating YBX1 in granulosa cells increased SIRT1 protein expression,SIRT1 mRNA stabili-ty,EdU positive rate,cell viability,PCNA mRNA expression level,Bcl2/BAX ratio(P＜0.05),while decreased Caspase-3 mRNA expression level,TUNEL positive rate(P＜0.05).Conclusions:The expression levels of YBX1 and SIRT1 in POI patients were significantly reduced and correlated with clinical ovarian function indicators.YBX1 can bind to SIRT1 mRNA and enhance its stability,which may promote the proliferation of granulosa cells,and in-hibit apoptosis.These findings suggested that YBX1 and SIRT1 are expected to become new targets for POI diag-nosis and treatment.

Objective:To investigate the effects of the Y-box binding protein 1(YBX1)on the proliferation and apoptosis of granulosa cells by regulating silent information regulator 1(SIRT1),and explore the differential ex-pression of YBX1 and SIRT1 of premature ovarian insufficiency(POI)patients and health controls,as well as its clinical significance.Methods:We recruited patients with POI and patients with normal ovarian function during in vitro fertilization(IVF)/intracytoplasmic sperm injection(ICSI)assisted pregnancy in the Department of Reproduc-tive Medical Center of the Fourth Affiliated Hospital of Jiangsu University from June 2022 to July 2023.The granu-losa cells and serum were collected.Western-Blot(WB)to detect YBX1 protein expression.Real-time quantitative polymerase chain reaction(RT-qPCR)to detect YBX1 and SIRT1 mRNA levels,and the correlation between the abundance of YBX1 and SIRT1 in serum and follicle stimulating hormone(FSH),luteinizing hormone(LH),estra-diol(E2),anti-Mullerian Hormone(AMH),and antral follicle count(AFC)were analyzed.5-bromo-2-deoxyuracil(EdU)and cell counting Kit-8(CCK8)were applied to detect cell proliferation;TdT-mediated dUTP-biotin nick end labeling assay(TUNEL)to detect cell apoptosis;RT-qPCR to detect cell proliferating cell nuclear antigen(PC-NA),B cell lymphoma 2(Bcl2),Bcl2 associated X protein(BAX),and cysteine-aspartate protease 3(Caspase-3)mRNA expression;RNA Immunoprecipitation(RIP)experiment to detect the interaction between YBX1 and SIRT1.Results:The expression of YBX1 protein and SIRT1 mRNA in granulosa cells and serum of POI patients were significantly lower than that of the control group(P＜0.05).The expression of YBX1 and SIRT1 mRNA in serum were negatively correlated with FSH,LH(r＜0,P＜0.05),and positively correlated with E2,AMH and AFC(r＞0,P＜0.05).Upregulating YBX1 in granulosa cells increased SIRT1 protein expression,SIRT1 mRNA stabili-ty,EdU positive rate,cell viability,PCNA mRNA expression level,Bcl2/BAX ratio(P＜0.05),while decreased Caspase-3 mRNA expression level,TUNEL positive rate(P＜0.05).Conclusions:The expression levels of YBX1 and SIRT1 in POI patients were significantly reduced and correlated with clinical ovarian function indicators.YBX1 can bind to SIRT1 mRNA and enhance its stability,which may promote the proliferation of granulosa cells,and in-hibit apoptosis.These findings suggested that YBX1 and SIRT1 are expected to become new targets for POI diag-nosis and treatment.