Characterization of two rat models of cystic fibrosis—KO and F508del CFTR—Generated by Crispr-Cas9

Background: Genetically engineered animals are essential for gaining a proper understanding of the disease mechanisms of cystic fibrosis(CF). The rat is a relevant laboratory model for CF because of its zootechnical capacity, size, and airway characteristics, including the presence of submucosal glands.Methods: We describe the generation of a CF rat model(F508 del) homozygous for the p.Phe508 del mutation in the transmembrane conductance regulator(Cftr) gene. This model was compared to new Cftr-/-rats(CFTR KO). Target organs in CF were examined by histological staining of tissue sections and tooth enamel was quantified by micro-computed tomography. The activity of CFTR was evaluated by nasal potential difference(NPD) and short-circuit current measurements. The effect of VX-809 and VX-770 was analyzed on nasal epithelial primary cell cultures from F508 del rats.Results: Both newborn F508 del and Knock out(KO) animals developed intestinal obstruction that could be partly compensated by special diet combined with an osmotic laxative. The two rat models exhibited CF phenotypic anomalies such as vas deferens agenesis and tooth enamel defects. Histology of the intestine, pancreas, liver, and lungs was normal. Absence of CFTR function in KO rats was confirmed ex vivo by short-circuit current measurements on colon mucosae and in vivo by NPD, whereas residual CFTR activity was observed in F508 del rats. Exposure of F508 del CFTR nasal primary cultures to a combination of VX-809 and VX-770 improved CFTR-mediated Cl-transport.Conclusions: The F508 del rats reproduce the phenotypes observed in CFTR KO animals and represent a novel resource to advance the development of CF therapeutics.

Genome-wide differences in hepatitis C-vs alcoholism-associated hepatocellular carcinoma

AIM:To look at a comprehensive picture of etiology- dependent gene abnormalities in hepatocellular carcinoma in Western Europe. METHODS:With a liver-oriented microarray,transcript levels were compared in nodules and cirrhosis from a training set of patients with hepatocellular carcinoma (alcoholism,12;hepatitis C,10)and 5 controls.Loose or tight selection of informative transcripts with an abnormal abundance was statistically valid and the tightly selected transcripts were next quantified by qRTPCR in the nodules from our training set(12+10) and a test set(6+7). RESULTS:A selection of 475 transcripts pointed to significant gene over-representation on chromosome 8 (alcoholism)or-2(hepatitis C)and ontology indicated a predominant inflammatory response(alcoholism)or changes in cell cycle regulation,transcription factors and interferon responsiveness(hepatitis C).A stringent selection of 23 transcripts whose differences betweenetiologies were significant in nodules but not in cirrhotic tissue indicated that the above dysregulations take place in tumor but not in the surrounding cirrhosis.These 23 transcripts separated our test set according to etiologies. The inflammation-associated transcripts pointed to limited alterations of free iron metabolism in alcoholic vs hepatitis C tumors. CONCLUSION:Etiology-specific abnormalities(chromo- some preference;differences in transcriptomes and related functions)have been identified in hepatocellular carcinoma driven by alcoholism or hepatitis C.This may open novel avenues for differential therapies in this disease.

面部肉芽肿:66例患者的临床病理学研究

Background: Only case reports or studies of small series of patients have focused on granuloma faciale (GF). Objective: We sought to describe the clinicopathologic characteristics ofGF in a large series of patients. Methods: We conducted a retrospective analysis of 66 patients and 73 skin specimens. Results: GF mostly presented as reddish plaques or nodules in middle-aged adults. One third of patients had multiple sites involved and 5 patients had extrafacial lesions. A clinical diagnosis of GF was made in only 10 cases; sarcoidosis, lymphoma, lupus, and basal cell carcinoma were the main differential diagnoses. The most frequent histopathologic features were the presence of a grenz zone, neutrophils, and telangiectases. Vascular changes were frequent, although necrotizing vasculitis appeared to be rare. There was often an association of acute and chronic inflammatory patterns, suggesting that GF follows a chronic history with reccurrent acute phases, rather than distinct successive acute and chronic stages. Limitations: In this retrospective study, a reliable analysis of the outcome of patients could not be performed. Results of direct immunofluorescence tests and laboratory investigations were lacking in many cases. Conclusion: GF is often clinically misdiagnosed; its morphologic spectrum is broader than usually described and includes a lack of eosinophils, the presence of vascular changeswith rare vessel wall necrosis, and associated acute and chronic inflammatory patterns.

接种疫苗后淋巴细胞增生肉芽肿10例(法语)

Background. Few cases of cutaneous lymphocytic hyperplasia secondary to vaccin ation have been published, although such lesions are not rare. Patients and meth ods. We report a series of 10 cases registered between 1993 and 2003. Results. M ean age was 25. The clinical aspect was solitary or multiple subcutaneous nodule s, located on the arm, developing after a delay of 1 to 18 months after vaccinat ion. Histologic examination showed a lymphocytic infiltration of the subcutaneou s fat, with diffuse and/or follicular pattern, without nuclear atypia, the morph ological and immunohistochemical analysis of which revealed the benigh nature. I n all cases, there was fibrosis and granuloma composed of lymphocytes, plasma ce lls, eosinophils and macrophages with basophilic cytoplasm. Morin stain showed i ntralesional aluminium in the 6 investigated cases. Evolution was always benign, with no relapse following exeresis. Discussion. Cutaneous lymphocytic hyperplas ia secondary to vaccination has to be suspected in a young patient with subcutan eous nodules appearing at a vaccination site. Evidence of aluminium in the lesio ns supports the diagnosis and the hypothesis that aluminium in the vaccine excip ient might have a role in the onset of such lesions.

Phylogenetic analysis of avian infectious bronchitis virus isolates from Morocco:a retrospective study(1983 to 2014)

Dear Editor,Infectious bronchitis(IB),one of the most common and difficult poultry diseases,is caused by a gammacoronavirus named infectious bronchitis virus(IBV).IBV frequently causes respiratory and/or renal diseases in chickens and egg production losses in hens.IB has

Dual angiogenesis and PD-1 blockade in hepatocellular carcinoma

Hepatocellular carcinoma(HCC)is the fourth leading cause of cancer related-death worldwide and its incidence is rising in most western countries(1).