OBJECTIVE:To study the effects of scorpion’s proteins with different molecular weights on angiogenesis of the transgenic zebrafish. METHODS:The vascular fluorescence transgenic zebrafish models were established. Scorpion’s proteins were separated by ultrafiltration and ion exchange chromatography to obtain the scorpion protein fractions with different molecular weights (3-10 ku,>10-50 ku and>50 ku). The embryos of transgenic zebrafishes were cultured in the above 10,100 and 500 μg/ml scor-pion’s proteins. Intersegmental vessels of the transgenic zebrafishes were counted under the fluorescence microscope to optimize the most suitable scorpion’s protein molecular weight. The vessels were counted again with >50 ku scorpion protein component 1 and 2,so as to select suitable component.RESULTS:The >50 ku scorpion’s protein fraction component 1 with the mass concentration of 500 μg/ml had the highest inhibitory activity for the angiogenesis of the transgenic zebrafish,with inhibitory rate of 92.59%. CONCLUSIONS:Scorpion’s protein and its fractions have the activity of angiogenesis inhibition,which may be one of anti-cancer mechanisms of scorpion.

Objective To investigate effects of hyperpolarization-activated cyclic nucleotide-gated cation nonselective channel in the hu-man ureter on the spontaneous contraction of smooth muscles. Methods Four HCN subtypes were detected in human ureteral tissue using reverse transcription polymerase chain reaction,Western blotting and immunohistochemical. ZD7288,the HCN blocker, was used to observe the changes of ureteral muscle contraction amplitude and frequency by applying the ureteral smooth muscle strip test in vitro. Results HCN1-4 isoforms were all identified in human ureter using reverse transcription-polymerase chain reaction and Western blotting. Through the immunohistochemical,HCN channel was found mostly in the urothelium layer and muscular layer of human ureteral wall. ZD7288 significantly decreased the bladder excitation. Conclusion All 4 HCN channel hypotypes exist in the human ureter, and affect the ureteral excitation.

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*

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 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.