OBJECTIVE: To explore the laboratory phenotype and molecular pathogenesis in a Chinese pedigree affected with Hereditary coagulation factor Ⅻ (FⅫ) deficiency. METHODS: A male proband admitted to Ningbo No.2 Hospital on July 17, 2021 due to chronic gastritis and members of his pedigree (7 individuals from three generations) were selected as the study subjects. Prothrombin time (PT), activated partial thromboplastin time (APTT), FⅧ activity (FⅧ: C), FⅨ activity (FⅨ: C), FⅪ activity (FⅪ: C), FⅫ activity (FⅫ: C), and FⅫ antigen (FⅫ: Ag) were determined. All of the exons, exon-intronic boundaries, as well as the 5'- and 3'-untranslated regions of the F12 gene were subjected to Sanger sequencing. Candidate variants were verified by cloning sequencing. The effect of candidate variants on the protein function was analyzed by bioinformatics software. RESULTS: The proband, a 47-year-old male, had significantly prolonged APTT (180.0 s) and decreased FⅫ:C and FⅫ:Ag levels (< 1%). His father, mother, brother and two sons also showed certain degrees of reduction. Genetic testing revealed that the proband has harbored compound heterozygous variants of the F12 gene, namely c.1092_1093insC (p.Lys365Glnfs*69) in exon 10 and c.1792_1796delGTCTA (p.Val579Hisfs*32) in exon 14. His mother and elder son were heterozygous for the c.1092_1093ins variant, whilst his father, brother, and younger son were heterozygous for the c.1792_1796delGTCTA variant. Analysis of the promoter region of exon 1 also showed that the proband and both sons had harbored a 46T/T polymorphism, whilst other family members were 46C/T. Bioinformatic analysis suggested that the p.Val579 is a highly conserved site. Protein model analysis showed that, with the p.Val579Hisfs*32 variant, a benzene ring was added and the hydrogen bond of surrounding amino acids was changed. Based on the guidelines from the American College of Medical Genetics and Genomics, the c.1792_1796delGTCTA was classified as a pathogenic variant (PVS1+PM2_Supporting+PM4). CONCLUSION The c.1092_1093insC (p.Lys365Glnfs*69) and c.1792_1796delGTCTA (p.Val579Hisfs*32) compound heterozygous variants of the F12 gene probably underlay the decreased FXII levels in this pedigree. Above finding has also enriched the mutational spectrum for FⅫ deficiency.
Male ; Humans ; Aged ; Middle Aged ; Pedigree ; East Asian People ; Exons ; Introns ; Family ; Factor XII Deficiency/genetics* ; 3' Untranslated Regions ; Factor XII/genetics*

OBJECTIVETo compare sub-acute toxic effects of cinnabar and Wansheng Huafeng Dan with mercury chloride and methyl-mercury.

METHODHealthy SD rats were orally administered with Wansheng Huafeng Dan (0.42 g x kg(-1)), cinnabar (0.15 g x kg(-1)), HgS (0.15 g x kg(-1)), HgCl2 (0.02 g x kg(-1)), MeHg (0.001 g x kg(-1)) and saline for 21 days under observed and their weights were monitored. After the final administration, they were decapitated and their blood, liver, kidney and brain tissues were collected for calculating hepatic and renal indexes and detecting the contents of serum glutamic pyruvic transaminase, urea nitrogen and creatinine and the mercury accumulation in liver, kidney and brain tissues. Besides, relative expressions of liver metallothionein-1 (MT-1) and cytochrome P450 gene subtypes (Cyp1a1, Cyp2b1, Cyp2e1, Cyp3a2, Cyp4a10) mRNA.

RESULTHgCl2 caused obvious weight lose in rats. Mercury contents in liver and kidney were markedly increased by HgCl2 and MeHg, and MeHg markedly increased mercury contents of brain either, but these advent effects were not notable in Wansheng Huafeng Dan and cinnabar groups. However, blood biochemistry and histopathology did not show significant changes in all groups. The expression of rat hepatic MT-1 mRNA was remarkably induced by both HgCl2 and MeHg. The expression of hepatic Cyp3a2 was increased by Wansheng Huafeng Dan and cinnabar, while the expression of Cyp2e1 was inhibited by HgCl2 and MeHg.

CONCLUSIONThe administration of Wansheng Huafeng Dan with equivalent dose for three weeks shows a much low sub-acute toxicity than HgCl2 and MeHg in rats.

Administration, Oral ; Animals ; Brain ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Kidney ; drug effects ; Liver ; drug effects ; Male ; Mercuric Chloride ; toxicity ; Mercury Compounds ; toxicity ; Methylmercury Compounds ; toxicity ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo detect numerical chromosomal aberrations and copy number variations (CNVs) in abortus tissue samples with a benchtop semiconductor sequencing platform (SSP).

METHODSOne hundred and forty patients with early spontaneous abortions at between 7 and 16 weeks were selected. Following DNA extraction, library preparation, high-throughput sequencing was carried out.

RESULTSChromosomal anomalies were detected in 82 cases (58.57%), which included 67 aneuploidies and 15 CNVs. Trisomies 16, 21, 22, and X/Y were the most common. A significant difference was found in the anomaly rates between women <35 and ≥35 years old (P< 0.01).

CONCLUSIONThe SSP platform can detect chromosomal aneuploidies and CNVs among abortus tissues. Numerical chromosomal aberration is the main cause of early spontaneous abortion, and advance maternal age is a high risk factor for chromosomal abnormality.

Objective To explore the clinical efficacy of using rituximab and atoxiban in the treatment of cervical incompetence after emergency cervical cerclage surgery.Methods Sixty patients with cervical incompetence admitted from May 2020 to February 2022 were selected as the study subjects.Divide into control group A,control group B,and trial group C using random number table method,with 20 cases in each group.All three groups underwent emergency cervical cerclage surgery.After the surgical treatment,control group A received treatment with rituximab hydrochloride,control group B received treatment with atoxiban,and experimental group C received treatment with atoxiban on the basis of rituximab hydrochloride.Analyze and compare the inhibitory effects of uterine contractions,incidence of adverse reactions,and neonatal outcomes among three groups.Results The inhibition rate of uterine contractions in control group A and test group C was higher than that in control group B(P<0.05);The incidence of postoperative complications in experimental group C was lower than that in control group A and control group B,with a statistically significant difference(P<0.05);Compared with control group A and control group B,the incidence of fetal loss,neonatal asphyxia,and low birth weight infant outcomes in experimental group C was not statistically significant(P>0.05).Compared with control group A and control group B,experimental group C had a longer gestational week extension and a higher Apgar score for newborns,with statistically significant differences(P<0.05).Conclusion The combination treatment of rituximab and atoxiban after emergency cervical cerclage surgery for this disease has good effects,can better inhibit uterine contractions,appropriately prolong gestational age,improve neonatal outcomes and prognosis,and reduce adverse drug reactions in pregnant women.

Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels are confirmed to be expressed in bladder interstitial Cajal-like cells (ICC-LCs), but little is known about their possible role in cystitis-associated bladder dysfunction. The present study aimed to determine the functional role of HCN channels in regulating bladder function under inflammatory conditions. Sixty female wild-type C57BL/6J mice and sixty female HCN1-knockout mice were randomly assigned to experimental and control groups, respectively. Cyclophosphamide (CYP)-induced cystitis models were successfully established in these mice. CYP treatment significantly enhanced HCN channel protein expression and I(h) density and significantly altered bladder HCN1 channel regulatory proteins. Carbachol (CCH) and forskolin (FSK) exerted significant effects on bladder ICC-LC [Ca²⁺]i in CYP-treated wild-type (WT) mice, and HCN1 channel ablation significantly decreased the effects of CCH and FSK on bladder ICC-LC [Ca²⁺]i in both naive and CYP-treated mice. CYP treatment significantly potentiated the spontaneous contractions and CCH (0.001-10 µM)-induced phasic contractions of detrusor strips, and HCN1 channel deletion significantly abated such effects. Finally, we demonstrated that the development of CYP-induced bladder overactivity was reversed in HCN1 -/- mice. Taken together, our results suggest that CYP-induced enhancements of HCN1 channel expression and function in bladder ICC-LCs are essential for cystitis-associated bladder hyperactivity development, indicating that the HCN1 channel may be a novel therapeutic target for managing bladder hyperactivity.
Animals ; Carbachol ; Colforsin ; Cyclophosphamide ; Cystitis ; Female ; Humans ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels* ; Mice* ; Telocytes* ; Up-Regulation* ; Urinary Bladder*

OBJECTIVE： To study early toxicity of paracetamol （APAP） to drug-induced liver injury in mice based on lipid metabonomics research， and to provide reference for finding potential biological marker. METHODS： Totally 20 mice were randomly divided into normal group and APAP liver injury group， with 10 mice in each group. APAP liver injury group was given intraperitoneal injection of APAP 300 mg/kg to establish acute liver injury model； normal group was given constant volume of normal saline intraperitoneally. 1 h later， the blood of mice was collected to isolate plasma. UPLC-Triple-TOF-MS method was used to detect plasma metabolites and perform metabonomics analysis. PCA， PLS-DA and OPLS-DA analysis distinguished the difference of metabolism profiles between groups. The lipid metabolites were screened and identified according to HMDB， Metlin and LIPID MAPS databases. Meanwhile， the changes of APAP level in plasma of mice were detected. The lipid metabolites with variable influence in the projection （VIP） greater than 1 and P＜0.05 in OPLS-DA analysis were identified as differential metabolites. The correlation between lipid differential metabolites and plasma APAP level was analyzed. RESULTS： PCA， PLS-DA and OPLS-DA results showed that sample points in normal group and APAP liver injury group were located in different areas with good differentiation. Compared with liver injury group and normal group， levels of 5 fatty acid metabolites were significantly increased or decreased； levels of 8 glycerophospholipids were significantly decreased and one sphingolipids was significantly increased. 9-thiastearic acid， tetradecanedioic acid， 9-hydrogen peroxide-10，12-octadecadienoic acid， L-myristoyl carnitine （fatty acid） and scyphostation A （sphingolipids） levels had a significant correlation with APAP level in plasma. CONCLUSIONS： The plasma lipid metabolomics showed abnormal changes 1 hour after acetaminophen exposure. A total of 14 related lipid differential metabolites are found， and 5 of which are significantly correlated with APAP level in plasma.

Objective:To observe the clinical features of uveal metastases from lung carcinoma.Methods:A retrospective case study. From 1983 to 2014, 14 patients with uveal metastases of lung cancer confirmed by ocular examination in Peking Union Medical College Hospital were included in the study. Among them, 7 were male, 7 were female; 11 were monocular and 3 were binocular. The mean age was 54.5±9.6 years. Pathologic examination showed primary bronchial lung cancer, including 13 patients of non-small cell lung cancer (10, 2 and 1 patients of lung adenocarcinoma, squamous cell carcinoma and adenosquamous cell carcinoma, respectively) and 1 patient of small cell lung cancer. Four patients (28.6%) were diagnosed with lung cancer before ophthalmology consultation, and 10 patients (71.4%) were first diagnosed with ophthalmology due to ocular symptoms. The duration from ocular symptoms to lung cancer diagnosis was 1 week to 6 months. The course from diagnosis of lung cancer to ophthalmological consultation was ranged from 10 to 60 months, and the average course was 29.5±19.0 months. There were 7, 4 and 3 patients with impaired vision, occlusion of visual objects and deformation of visual objects, respectively. All patients underwent visual acuity, slit lamp microscope, B-mode ultrasound and UBM examinations. FFA was performed in 8 eyes, and 2 eyes were examined for ICGA. Orbital MRI was performed in 5 patients. Vitreoretinal surgery was performed on 1 eye. The clinical characteristics of the patients were analyzed and observed.Results:In 17 eyes, there were 2 eyes with visual acuity of light perception, 3 eyes of hand movement to counting finger before the eyes, 5 eyes of 0.1- 0.3, 4 eyes of 0.4-0.6, 3 eyes of greater than 0.8. Metastatic cancer was located in iris in 1 eye, it presents as a red mass with irregular shape on the surface, which is full of small nourishing blood vessels. Metastatic cancer were located in choroid in 16 eyes, they presented yellowish-white or grayish-yellow lumps under the posterior pole or equatorial retina, including 14 eyes with a single lesion and 2 eyes with 2 lesions, with retinal detachment in 8 eyes and increased intraocular pressure in 5 eyes. B-mode ultrasonography showed posterior polar flat or surface irregular wavy intraocular space occupying lesions with localized or extensive retinal detachment. FFA and ICGA showed the focal, apical and patchy fluorescence of the tumor. MRI showed that T 1WI medium and high signal consistent with the vitreous body, while T 2WI showed low signal. Conclusions:Uveal metastatic may be the first manifestation of lung cancer, and visual impairment, part of solid mass lesions with fundus flattening may be accompanied by secondary glaucoma and retinal detachment as the main clinical manifestations. Most of the metastatic sites are located in choroid, which is more common in single eye and single lesion. Adenocarcinoma is the most common type of uveal metastasis in non-small cell lung cancer.

OBJECTIVE To optimite the purification technology of total triterpenoid extracts from Inonotus obliquus ，and to investigate the anti -tumor activity of its purified products . METHODS Using inotodiol as control ，the method was established for the content determination of total triterpenoid in I. obliquus. The type of macroporous adsorption resin ，sample volume ，sample concentration，sample flow rate ，eluent volume ，eluent dosage and elution flow rate were selected by single factor experiments . The purification technology of the crude extract was determined and verified . The effects of total triterpenoid purified from I. obliquus on the proliferation ，migration and apoptosis of human cervical cancer HeLa cells were detected by cell proliferation test ， migration test ，flow cytometry and AO/EB kit . RESULTS The best purification technology of total triterpenoid crude extracts from I. obliquus was as follows ：AB-8 macroporous adsorption resin was used ；mass concentration of the sample solution was 2.0 mg/mL；sample volume was 140 mL，and the flow rate was 1.0 mL/min；the impurity was removed with 50% ethanol 40 mL， then eluted with 95% ethanol 160 mL，at the elution flow rate of 3.0 mL/min. After purification ，mass concentration of total triterpenoid from I. obliquus increased from 34.36% to 73.39%. The total triterpenoid of I. obliquus could inhibit the proliferation of HeLa cells ，and the 50% inhibitory concentration was 184.20 μg/mL. Compared with control group ，the purified products could significantly inhibit the migratio n and promote the apoptosis of HeLa cells （P＜0.05 or P＜0.01）. CONCLUSIONS The purification technology of total triterpenoids extracts from I. obliquus is successfully optimited . The purified product could inhibit the proliferation and migration of HeLa cells and induce their apoptosis.