Given that ovarian stimulation is vital for assisted reproductive technology (ART) and results in elevated serum estrogen levels, exploring the impact of elevated estrogen exposure on oocytes and embryos is necessary. We investigated the effects of various ovarian stimulation treatments on oocyte and embryo morphology and gene expression using a mouse model and estrogen-treated mouse embryonic stem cells (mESCs). Female C57BL/6J mice were subjected to two types of conventional ovarian stimulation and ovarian hyperstimulation; mice treated with only normal saline served as controls. Hyperstimulation resulted in high serum estrogen levels, enlarged ovaries, an increased number of aberrant oocytes, and decreased embryo formation. The messenger RNA (mRNA)‍-sequencing of oocytes revealed the dysregulated expression of lysine-specific demethylase 6b (Kdm6b), which may be a key factor indicating hyperstimulation-induced aberrant oocytes and embryos. In vitro, Kdm6b expression was downregulated in mESCs treated with high-dose estrogen; treatment with an estrogen receptor antagonist could reverse this downregulated expression level. Furthermore, treatment with high-dose estrogen resulted in the upregulated expression of histone H3 lysine 27 trimethylation (H3K27me3) and phosphorylated H2A histone family member X (γ-H2AX). Notably, knockdown of Kdm6b and high estrogen levels hindered the formation of embryoid bodies, with a concomitant increase in the expression of H3K27me3 and γ-H2AX. Collectively, our findings revealed that hyperstimulation-induced high-dose estrogen could impair the demethylation of H3K27me3 by reducing Kdm6b expression. Accordingly, Kdm6b could be a promising marker for clinically predicting ART outcomes in patients with ovarian hyperstimulation syndrome.
Female ; Mice ; Demethylation/drug effects* ; Embryonic Stem Cells ; Estrogens/administration & dosage* ; Gene Expression/drug effects* ; Histones/metabolism* ; Jumonji Domain-Containing Histone Demethylases/metabolism* ; Mice, Inbred C57BL ; Oocytes ; Ovary/drug effects* ; Reproductive Techniques, Assisted ; Animals

BACKGROUND: B-cell maturation antigen (BCMA)-directed chimeric antigen receptor T (CAR-T) therapy yield remarkable responses in patients with relapsed/refractory multiple myeloma (R/RMM). Circulating tumor DNA (ctDNA) reportedly exhibits distinct advantages in addressing the challenges posed by tumor heterogeneity in the distribution and genetic variations in R/RMM. METHODS: Herein, the ctDNA of 108 peripheral blood plasma samples from patients with R/RMM was thoroughly investigated before administration of anti-BCMA CAR-T therapy to establish its predictive potential. Flow cytometry is used primarily to detect subgroups of T cells or CAR-T cells. RESULTS: In this study, several tumor and T cell effector-mediated factors were considered to be related to treatment failure by an integrat analysis, including higher percentages of multiple myeloma (MM) cells in the bone marrow (P = 0.013), lower percentages of CAR-T cells in the peripheral blood at peak (P = 0.037), and higher percentages of CD8+ T cells (P = 0.034). Furthermore, there is a substantial correlation between high ctDNA level (>143 ng/mL) and shorter progression-free survival (PFS) (P = 0.007). Multivariate Cox regression analysis showed that high levels of ctDNA (>143 ng/mL), MM-driven high-risk mutations (including IGLL5 [P = 0.004], IRF4 [P = 0.024], and CREBBP [P = 0.041]), number of multisite mutations, and resistance-related mutation (ERBB4, P = 0.040) were independent risk factors for PFS. CONCLUSION: Finally, a ctDNA-based risk model was built based on the above independent risk factors, which serves as an adjunct non-invasive measure of substantial tumor burden and a prognostic genetic feature that can assist in predicting the response to anti-BCMA CAR-T therapy. REGISTERATION Chinese Clinical Trial Registry (ChiCTR2100046474) and National Clinical Trial (NCT04670055, NCT05430945).

Objective:To evaluate the feasibility and clinical value of pre-treatment non-enhanced chest CT radiomics features and machine learning algorithm to predict the mutation status and subtype (19Del/21L858R) of epidermal growth factor receptor (EGFR) for patients with non-small cell lung cancer (NSCLC).Methods:This retrospective study enrolled 280 NSCLC patients from first and second affiliated hospital of University of South China who were confirmed by biopsy pathology, gene examination, and have pre-treatment non-enhanced CT scans. There are 136 patients were confirmed EGFR mutation. Primary lung gross tumor volume was contoured by two experienced radiologists and oncologists, and 851 radiomics features were subsequently extracted. Then, spearman correlation analysis and RELIEFF algorithm were used to screen predictive features. The two hospitals were training and validation cohort, respectively. Clinical-radiomics model was constructed using selected radiomics and clinical features, and compared with models built by radiomics features or clinical features respectively. In this study, machine learning models were established using support vector machine (SVM) and a sequential modeling procedure to predict the mutation status and subtype of EGFR. The area under receiver operating curve (AUC-ROC) was employed to evaluate the performances of established models.Results:After feature selection, 21 radiomics features were found to be efffective in predicting EGFR mutation status and subtype and were used to establish radiomics models. Three types models were established, including clinical model, radiomics model, and clinical-radiomics model. The clinical-radiomics model showed the best predictive efficacy, AUCs of predicting EGFR mutation status for training dataset and validation dataset were 0.956 (95% CI: 0.952-1.000) and 0.961 (95% CI: 0.924-0.998), respectively. The AUCs of predicting 19Del/L858R mutation subtype for training dataset and validation dataset were 0.926 (95% CI: 0.893-0.959), 0.938 (95% CI: 0.876-1.000), respectively. Conclusions:The constructed sequential models based on integration of CT radiomics, clinical features and machine learning can accurately predict the mutation status and subtype of EGFR.

Aconitum kusnezoffii is a traditional Chinese medicine of Ranunculaceae family. Its toxicity is relatively strong, and its dosage is similar to that of poisoning. In clinical practice, poisoning events are often caused by excessive dosage or improper use. There is no specific antidote for kusnezoff root poisoning. Severe kusnezoff root poisoning can cause malignant arrhythmia and even death.A case of severe kusnezoff monkshood poisoning was reported in January 2021, which was treated with nificaran hydrochloride for injection in the emergency medicine department of the First Hospital of Handan City. The patient developed ventricular tachycardia, ventricular fibrillation and AS syndrome. In addition to conventional treatment, the patient did not have arrhythmia again after intravenous injection of 25 mg of nifekalan load and continuous pumping of 0.4 mg/kg/h for 7 hours, and did not relapse after discontinuation of nifekalan 24 hours later. It is suggested that the malignant arrhythmia caused by clinical severe kusnezoff monkshood poisoning can be controlled by nifekalan. Whether nifekalan is superior to conventional antiarrhythmic drugs still needs more accumulation and verification of clinical application data.

Aconitum kusnezoffii is a traditional Chinese medicine of Ranunculaceae family. Its toxicity is relatively strong, and its dosage is similar to that of poisoning. In clinical practice, poisoning events are often caused by excessive dosage or improper use. There is no specific antidote for kusnezoff root poisoning. Severe kusnezoff root poisoning can cause malignant arrhythmia and even death.A case of severe kusnezoff monkshood poisoning was reported in January 2021, which was treated with nificaran hydrochloride for injection in the emergency medicine department of the First Hospital of Handan City. The patient developed ventricular tachycardia, ventricular fibrillation and AS syndrome. In addition to conventional treatment, the patient did not have arrhythmia again after intravenous injection of 25 mg of nifekalan load and continuous pumping of 0.4 mg/kg/h for 7 hours, and did not relapse after discontinuation of nifekalan 24 hours later. It is suggested that the malignant arrhythmia caused by clinical severe kusnezoff monkshood poisoning can be controlled by nifekalan. Whether nifekalan is superior to conventional antiarrhythmic drugs still needs more accumulation and verification of clinical application data.

OBJECTIVE: To assess the association of CYP2C19 and CYP3A5 gene polymorphisms with the risk of myocardial infarction. METHODS: Five hundred patients with myocardial infarction and 500 healthy controls were randomly selected. Fluorescent PCR and Sanger sequencing were used to detect the CYP2C19 and CYP3A5 gene polymorphisms. Logistic regression was used to analyze the correlation between the polymorphisms and myocardial infarction. Quanto software was used to evaluate the statistical power. RESULTS: The two groups had significant difference in the frequency of AG, GG genotypes and A allele of the CYP2C19 gene rs4986893 locus and the AA, AG, GG genotypes and G allele of the CYP3A5 gene rs776746 locus ( P<0.05), but not in the frequency of genotypes and alleles of CYP2C19 gene rs4244285 and rs12248560 loci, and the AA genotype of the rs4986893 locus. After correction for age, gender, and body mass index, Logistic regression indicated that the AG genotype and A allele of the CYP2C19 gene rs4986893 locus, and the GG genotype and G allele of CYP3A5 gene rs776746 locus are associated with susceptibility of myocardial infarction, while rs4986893 GG genotype and AA and AG genotypes of rs776746 may confer a protective effect. Based on the sample size and allele frequency, analysis with Quanto software suggested that the result of this study has a statistical power of 99%. CONCLUSION CYP2C19 and CYP3A5 gene polymorphisms may increase the risk for myocardial infarction.
Cytochrome P-450 CYP2C19/genetics* ; Cytochrome P-450 CYP3A/genetics* ; Gene Frequency ; Genotype ; Humans ; Myocardial Infarction/genetics* ; Polymorphism, Genetic ; Polymorphism, Single Nucleotide

Objective To analyze the clinical and biochemical,as well as genetic characteristics of a patient with Wiedemann-Steiner syndrome (WDSTS).Methods The clinical data of a patient with WDSTS were collected.The patient was treated with recombinant human growth hormone (rhGH) combined with gonadotrophine-releasing hormone agonist (GnRHa).Blood samples of the patient and her parents were taken for whole-Exome Sequencing (WES).Relevant literatures about KMT2A mutations were reviewed.Results The 5-year old girl presented with growth retardation,with height 100 cm (-2.4 SD),torpid reaction,and facial anomalies including low hairline,thick eyebrow and hair,hypertelorism,a wide nasal bridge.She had small and puffy hands and feet,excessive hair around back of neck,bilateral forearm and lower limbs.Her GH peak level was 26.6 ng/ml during GH stimulation test.She was re-examined at the age of 10.4 years,with severe short stature (120 cm/-3.58 SD) and a Tanner stage 2 of breast development.Her bone age was found to be approximately 11.4 years.Height increased from 120 cm at the age of 10.4 years to 147.3 cm after rhGH treatment combined with GnRHa for 2.5 years.rhGH therapy alone continued for 1.1 years and a height of 150 cm was reached at the age of 14.9 years,with bone age 14 years.Gene sequencing revealed a de novo frameshift mutation (c.10051 delA,p.Thr3351 Leufs * 17) of exon 27 in KMT2A gene of the patient,but without any mutation in her parents.Through a literature review,seventy-one patients with WDSTS (including present case) presented with intellectual disability (70/71),facial anomalies (70/71),short stature (50/71),and hypertrichosis (39/71).Conclusion Patients presented with short stature,typical facial dysmorphism,intellectual disability,and hypertrichosis should be considered for WDSTS.The mutation p.Thr3351Leufs * 17 in the KMT2A gene detected in our patient is a novel mutation.This is so far the first report of WDSTS patient who was successfully treated with a combination of GH and GnRHa at the onset of puberty to improve her adult height.

Objective To analysis the clinical features,diagnosis,treatment and prognosis of adrenal eosinophilic tumor with low testosterone levels.Methods The clinical data of a 22 years old male patient with adrenal eosinophilic tumor and low testosterone levels was analyzed.Blood pressure was 151/88 mmHg.The patient got bilateral gynecomastia.His bilateral testicular was soft and became smaller,with short penisr.Endocrine examination results showed:Estradiol 666 pg/ml,Prolactin 19.08 ng/ml,Testosterone 0.18 ng/ml,follicle stimulating hormone ＜ 0.2 U/L.The CT showed the mass density of soft tissue in the left adrenal region with diameter 7 cm,which was inhomogeneous and enhanced.There were many small vessels enhanced in the CT arterial phase,and the blood flow in the tumor was abundant.Clinical diagnosis of left adrenal tumor was pheochromocytoma.The patient underwent laparoscopic left adrenal tumor resection.The left adrenal gland was located in the superior pole of the left kidney,and there was an independent supply of the artery.Results Pathological result showed the tumor weigh was 60 g,7 cm in diameter and brown in section.The tumor cells were arranged in solid nests or acini,with more eosinophilic granules in cytoplasm.The nuclei was round and the nucleoli was located in the center,had clusters of pleomorphic and clustered cells.The tumor was wrapped in a thick fibrous envelope,mainly consisted of eosinophils,granulation tissue.There was no necrosis,mitosis,and vascular invasion.Immunohistochemical staining showed that the expression of CD56 and syn protein was positive.Pathological diagnosis was left adrenal eosinophilic tumor.After 4 months,the blood testosterone levels rose to 3.90 ng/ml,the blood pressure returned to normal (118/75 mmhg).The estradiol (21 pg/ml) was significantly inhibited.The patient began to appear beards and breasts became smaller.There were no signs of clinical or imaging recurrence.After 16 months follow-up,serum testosterone was 4.68 ng/ml and serum estrogen levels dropped to 33 pg/ml.Semen routine showed no sperm.Conclusions The clinical morbidity of functional adrenocortical oncocytoma with low testosterone levels and high estradiol levels is low.The pathological components are mainly eosinophilic granulation tissue.The adrenocortical oncocytoma are rare and preoperative diagnosis is difficult.Clinical manifestation,imaging examination and adrenal biochemistry examination should be considered to determine the localization and qualitative of tumor.Minimally invasive surgery is an effective treatment.The close follow-up after operation is essential.

Objective To establish a patient-derived xenografts (PDX) mouse model of liver cancer (LC) and to explore its role in precision medicine.Methods PDX model was established by subcutaneous implantation of tumor tissues in NCG mice.The morphological structure of tumor tissue was exaimed using HE staining.Fifteen BALB/c nude mice were subcutaneously inoculated with tumor cell suspension from the PDX models.The xenograft mice were randomly divided into 5-fluorouracil (5-FU) group, sorafenib group and negative control group.The tumor volume and body weight of the tumor-bearing mice were measured regularly, the tumor inhibition rate was calculated and the curative effect was evaluated.Results The success rate was 33.3% (6/18) in the establishment of liver cancer PDX mouse model, and the model well retained the characteristics of the primary tumor.In one case of PDX mouse model, the tumor inhibition rates of 5-FU and sorafenib group were 63.7% and 29.6%, with a statistically significant differece between them (P< 0.05), and there was no significant difference between the sorafenib group and negative control group, consistent with clinical observation.Conclusions The PDX mouse model of liver cancer can maintain the histological structure of primary tumor, and can be applied to precision medicine for patients with liver cancer.