Enfermedad hepática en la fibrosis quística pediátrica. Una revisión de los conocimientos actuales / Cystic fibrosis liver disease in children ­ A review of our current understanding

La fibrosis quística, la segunda enfermedad genética más frecuente, es el resultado de una proteína de canal mutada, la CFTR, que secreta iones de cloro que fluidifican las secreciones. La esperanza de vida en los pacientes ha aumentado en años recientes gracias a mejoras en el tratamiento. No obstante, las complicaciones hepáticas son la tercera causa de muerte y la comprensión de su fisiopatología es aún deficiente. Se considera que la obstrucción biliar secundaria a la presencia de secreciones espesas conduce a la cirrosis. Sin embargo, el ácido ursodesoxicólico no ha modificado la historia natural. Además, la presencia de hipertensión portal en ausencia de cirrosis no puede ser explicada. Se ha propuesto el rol de la CFTR como modulador de tolerancia inmune, que explica la presencia de una inflamación portal persistente que culmina en fibrosis. El eje intestino-hígado tendría un rol importante en la presentación y la progresión de esta enfermedad

Cystic fibrosis is the second most common genetic disease in infancy. It is the result of a mutated channel protein, the CFTR, which secretes chloride ions, fluidifying secretions. Recent improvements in the treatment have increased life expectancy in these patients. Nevertheless, liver involvement remains the third cause of death. Unfortunately, our understating of the physiopathology is still deficient. Biliary obstruction secondary to the presence of thick secretions is considered to lead to cirrhosis. However, treatment with ursodeoxycolic acid has not changed the natural history. Furthermore, the presence of portal hypertension in the absence of cirrhosis cannot be explained. Recently, the role of CFTR as modulator of immune tolerance has been proposed, which could explain the presence of a persistent portal inflammation leading to fibrosis, and the gut-liver axis would also have a role in disease presentation and progression.

Identification of risk genes in Chinese nonobstructive azoospermia patients based on whole-exome sequencing / 亚洲男科学杂志(英文版)

Nonobstructive azoospermia (NOA) is a severe condition in infertile men, and increasing numbers of causative genes have been identified during the last few decades. Although certain causative genes can explain the presence of NOA in some patients, a proportion of NOA patients remain to be addressed. This study aimed to investigate potential high-risk genes associated with spermatogenesis in idiopathic NOA patients by whole-exome sequencing. Whole-exome sequencing was performed in 46 male patients diagnosed with NOA. First, screening was performed for 119 genes known to be related to male infertility. Next, further screening was performed to determine potential high-risk causative genes for NOA by comparisons with 68 healthy male controls. Finally, risk genes with high/specific expression in the testes were selected and their expression fluctuations during spermatogenesis were graphed. The frequency of cystic fibrosis transmembrane conductance regulator (CFTR) gene pathogenic variant carriers was higher in the NOA patients compared with the healthy controls. Potential risk genes that may be causes of NOA were identified, including seven genes that were highly/specifically expressed in the testes. Four risk genes previously reported to be involved in spermatogenesis (MutS homolog 5 [MSH5], cilia- and flagella-associated protein 54 [CFAP54], MAP7 domain containing 3 [MAP7D3], and coiled-coil domain containing 33 [CCDC33]) and three novel risk genes (coiled-coil domain containing 168 [CCDC168], chromosome 16 open reading frame 96 [C16orf96], and serine protease 48 [PRSS48]) were identified to be highly or specifically expressed in the testes and significantly different in the 46 NOA patients compared with 68 healthy controls. This study on clinical NOA patients provides further evidence for the four previously reported risk genes. The present findings pave the way for further functional investigations and provide candidate risk genes for genetic diagnosis of NOA.

El regulador de transmembrana de fibrosis quística (CFTR) disfuncional y sus efectos extrapulmonares / The dysfunction of transmembrane regulator in cystic fibrosis (CFTR) and its extrapulmonary effects

El daño del regulador de transmembrana de fibrosis quística (CFTR) puede causar una enfermedad grave fuera de los pulmones. El canal de cloruro (Cl-) ha sido el más estudiado, sin embargo, el bicarbonato (HCO3 -) tiene un rol muy importante en el comportamiento de las secreciones y la inflamación secundaria. El hecho de que CFTR funcione no sólo como un canal de Cl- sino también de HCO3- es un campo para la investigación y el desarrollo de fármacos para pacientes con daño genético o adquirido, este último frecuente en la población general. Algunos moduladores de CFTR pueden tener un beneficio terapéutico en el tratamiento de pancreatitis en ambas situaciones. La disfunción del CFTR a nivel renal puede resultar excepcionalmente en alcalosis metabólica y reducción del impulso ventilatorio. Hasta la fecha no está claro cuales serian sus efectos en los sistemas gastrointestinal y hepatobiliar.

Transmembrane regulator in cystic fibrosis (CFTR) can cause severe disease outside of the lungs. The chloride channel (Cl-) has been the most studied, however bicarbonate (HCO3 -) has a very important role in the behavior of secretions and secondary inflammation. The fact that CFTR works not only as a Cl- channel but also as an HCO3- channel is a field for research and development of drugs for patients with genetic or acquired damage, the latter frequent in the general population. Some CFTR modulators may have a therapeutic benefit in the treatment of pancreatitis in both situations. CFTR dysfunction at the renal level can exceptionally result in metabolic alkalosis and reduced ventilatory drive. To date it is not clear what its effects on the gastrointestinal and hepatobiliary systems would be.

Actualización en fibrosis quística: diabetes relacionada a fibrosis quística en la era de los moduladores del regulador de transmembrana de fibrosis quística (CFTR) / Update on cystic fibrosis/cystic fibrosis related diabete in the era of transmembrane regulator in cystic fibrosis (CFTR)

Las disglicemias, objetivadas en el test de tolerancia a la glucosa de 2 horas y en el monitoreo continuo de glicemia, son el factor de riesgo principal para el desarrollo de la diabetes relacionada a fibrosis quística (FQ) (DRFQ), la que constituiría la etapa final de un continuo de alteraciones del metabolismo de la glucosa en los pacientes con FQ. Estas disglicemias se deben tanto al daño directo de las células de los islotes pancreáticos productores de insulina, como al aumento de la resistencia a la insulina asociada al estado inflamatorio sistémico de la FQ. El uso cada vez más precoz de los moduladores del CFTR debiera contribuir a evitar el desarrollo de DRFQ y sus complicaciones. La siguiente revisión se enfoca en los efectos de los moduladores del CFTR en la tolerancia a la glucosa en pacientes con FQ.

Dysglycemia, observed in the 2-hour glucose tolerance test and in the continuous monitoring of glycemia, are the main risk factor for the development of diabetes related to cystic fibrosis (CF), which constitutes the final stage of a continuum of impaired glucose metabolism in people with CF. These dysglycemias are due both to direct damage to insulin-producing pancreatic islet cells, and to increased insulin resistance associated with the systemic inflammatory state of CF. The increasingly early use of CFTR modulators should help prevent the development of CRFD and its complications. The following review focuses on the effects of regulador de transmembrana de fibrosis quística (CFTR) modulators on glucose tolerance in people with CF.

Use of elexacaftor+tezacaftor+ivacaftor in individuals with cystic fibrosis and at least one F508del allele: a systematic review and meta-analysis / Uso de elexacaftor + tezacaftor + ivacaftor em indivíduos com fibrose cística com pelo menos um alelo F508del: revisão sistemática e meta-análise

ABSTRACT Objective: To evaluate the effect of treatment with the combination of three cystic fibrosis transmembrane conductance regulator (CFTR) modulators-elexacaftor+tezacaftor+ivacaftor (ETI)-on important clinical endpoints in individuals with cystic fibrosis. Methods: This was a systematic review and meta-analysis of randomized clinical trials that compared the use of ETI in individuals with CF and at least one F508del allele with that of placebo or with an active comparator such as other combinations of CFTR modulators, following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) recommendations and the Patients of interest, Intervention to be studied, Comparison of interventions, and Outcome of interest (PICO) methodology. We searched the following databases: MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, and ClinicalTrials.gov from their inception to December 26th, 2022. The risk of bias was assessed using the Cochrane risk-of-bias tool, and the quality of evidence was based on the Grading of Recommendations Assessment, Development and Evaluation (GRADE). Results: We retrieved 54 studies in the primary search. Of these, 6 met the inclusion criteria and were analyzed (1,127 patients; 577 and 550 in the intervention and control groups, respectively). The meta-analysis revealed that the use of ETI increased FEV1% [risk difference (RD), +10.47%; 95% CI, 6.88-14.06], reduced the number of acute pulmonary exacerbations (RD, −0.16; 95% CI, −0.28 to −0.04), and improved quality of life (RD, +14.93; 95% CI, 9.98-19.89) and BMI (RD, +1.07 kg/m2; 95% CI, 0.90-1.25). Adverse events did not differ between groups (RD, −0.03; 95% CI, −0.08 to 0.01), and none of the studies reported deaths. Conclusions: Our findings demonstrate that ETI treatment substantially improves clinically significant, patient-centered outcomes.

RESUMO Objetivo: Avaliar o efeito do tratamento com a combinação de três moduladores da proteína cystic fibrosis transmembrane conductance regulator (CFTR, reguladora de condutância transmembrana em fibrose cística) - elexacaftor + tezacaftor + ivacaftor (ETI) - sobre desfechos clínicos importantes em indivíduos com fibrose cística. Métodos: Revisão sistemática e meta-análise de ensaios clínicos randomizados que compararam o uso de ETI em indivíduos com fibrose cística com pelo menos um alelo F508del com o uso de placebo ou de um comparador ativo como outras combinações de moduladores da CFTR. O estudo foi realizado seguindo as recomendações Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) e a metodologia Patients of interest, Intervention to be studied, Comparison of interventions, and Outcome of interest (PICO). Foram realizadas buscas nos seguintes bancos de dados: MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials e ClinicalTrials.gov, desde a sua criação até 26 de dezembro de 2022. O risco de viés foi avaliado por meio da ferramenta de risco de viés da Cochrane, e a qualidade das evidências foi determinada com base no sistema Grading of Recommendations Assessment, Development and Evaluation (GRADE). Resultados: Foram identificados 54 estudos na busca primária. Destes, 6 preencheram os critérios de inclusão e foram analisados (1.127 pacientes: 577 pacientes intervenção e 550 pacientes controle). A meta-análise revelou que o uso de ETI aumentou o VEF1 em porcentagem do previsto [diferença de risco (DR): +10,47%; IC95%: 6,88-14,06], reduziu o número de exacerbações pulmonares agudas (DR: −0,16; IC95%: −0,28 a −0,04) e melhorou a qualidade de vida (DR: +14,93; IC95%: 9,98-19,89) e o IMC (DR: +1,07 kg/m2; IC95%: 0,90-1,25). Os eventos adversos não diferiram entre os grupos (DR: −0,03; IC95%: −0,08 a 0,01), e nenhum dos estudos relatou óbitos. Conclusões: Nossos achados demonstram que o tratamento com ETI melhora substancialmente os desfechos clinicamente significativos centrados no paciente.

Elexacaftor/tezacaftor/ivacaftor-real-world clinical effectiveness and safety. A single-center Portuguese study / Elexacaftor/tezacaftor/ivacaftor-efetividade clínica e segurança no mundo real. Estudo unicêntrico português

ABSTRACT Objective: To evaluate the effectiveness of treatment with elexacaftor/tezacaftor/ivacaftor (ELX/TEZ/IVA) and to characterize its safety profile in cystic fibrosis (CF) patients in a real-world clinical setting. Methods: This was a prospective observational study carried out in a CF referral center in Portugal involving adult CF patients who started treatment with ELX/TEZ/IVA. Clinical characteristics of the patients were collected, and effectiveness and safety data were evaluated. Results: Of the 56 patients followed in the center at the time of the study, 28 were eligible for ELX/TEZ/IVA treatment in accordance with the Portuguese National Authority for Medicines and Health Products at the time of the study. Of these, 24 met the follow-up time requirement to be included in the clinical effectiveness analysis. The mean follow-up time was 167.3 ± 96.4 days. Adverse events were generally mild and self-limited. Significant improvements in lung function, BMI, sweat chloride concentration, and number of pulmonary exacerbations were observed. No significant differences in outcomes between F508del homozygous and heterozygous patients were found. The effectiveness of this new CFTR modulator combination also applied to patients with advanced lung disease. Conclusions: Treatment with ELX/TEZ/IVA showed effective improvement in real-world clinical practice, namely in lung function, BMI, sweat chloride concentration, and number of pulmonary exacerbations, with no safety concerns.

RESUMO Objetivo: Avaliar a efetividade do tratamento com elexacaftor/tezacaftor/ivacaftor (ELX/TEZ/IVA) e caracterizar seu perfil de segurança em pacientes com fibrose cística (FC) em um cenário clínico de mundo real. Métodos: Estudo observacional prospectivo realizado em um centro de referência em FC de Portugal com pacientes adultos com FC que iniciaram o tratamento com ELX/TEZ/IVA. As características clínicas dos pacientes foram coletadas, e os dados de efetividade e segurança, avaliados. Resultados: Dos 56 pacientes acompanhados no centro na época do estudo, 28 eram elegíveis para o tratamento com ELX/TEZ/IVA de acordo com a Autoridade Nacional do Medicamento e Produtos de Saúde. Destes, 24 atenderam ao requisito de tempo de acompanhamento para inclusão na análise de efetividade clínica. O tempo médio de acompanhamento foi de 167,3 ± 96,4 dias. Os eventos adversos foram geralmente leves e autolimitados. Foram observadas melhoras significativas na função pulmonar, no IMC, na concentração de cloreto no suor e no número de exacerbações pulmonares. Não foram encontradas diferenças significativas nos resultados entre os pacientes homozigotos e heterozigotos para F508del. A efetividade dessa nova combinação de moduladores da CFRT em fibrose cística também se aplica a pacientes com doença pulmonar avançada. Conclusões: O tratamento com ELX/TEZ/IVA demonstrou melhora efetiva na prática clínica real, a saber, na função pulmonar, no IMC, na concentração de cloreto no suor e no número de exacerbações pulmonares, sem preocupações de segurança.

Association between phenotypic and genotypic characteristics and disease severity in individuals with cystic fibrosis / Associação entre as características fenotípicas, genotípicas e a gravidade da doença em indivíduos com fibrose cística

Abstract Objective: To analyze the association between phenotypic and genotypic characteristics and disease severity in individuals with cystic fibrosis treated at a reference center in Minas Gerais, Brazil. Methods: This is a retrospective study that collected clinical and laboratory data, respiratory and gastrointestinal manifestations, type of treatment, Shwachman-Kulczycki score, and mutations from the patients' medical records. Results: The sample included 50 participants aged one to 33 years, 50% of whom were female. Out of the one hundred alleles of the Cystic Fibrosis Transmembrane Conductance Regulator gene, the most prevalent mutations were DeltaF508 (45%) and S4X (18%). Mutation groups were only associated with pancreatic insufficiency (p=0.013) and not with disease severity (p=0.073). The latter presented an association with colonization by Pseudomonas aeruginosa and Staphylococcus aureus (p=0.007) and with underweight (p=0.036). Death was associated with age at diagnosis (p=0.016), respiratory symptomatology (p=0.013), colonization (p=0.024), underweight (p=0.017), and hospitalization (p=0.003). Conclusions: We could identify the association of mutations with pancreatic insufficiency; the association of Staphylococcus aureus colonization and underweight with disease severity; and the lack of association between mutations and disease severity. Environmental factors should be investigated more thoroughly since they seem to have an important effect on disease severity.

RESUMO Objetivo: Analisar a associação entre as características fenotípicas, genotípicas e a gravidade da doença em indivíduos com fibrose cística atendidos em um centro de referência de Minas Gerais, Brasil. Métodos: Trata-se de um estudo retrospectivo, em que os dados clínicos, laboratoriais, as manifestações respiratórias e gastrointestinais, o tipo de tratamento, o escore de Shwachman-Kulczycki e as mutações foram coletados dos prontuários de registros dos pacientes. Resultados: A amostra incluiu 50 participantes, de um a 33 anos de idade, sendo 50% do sexo feminino. Do total de cem alelos do gene Cystic Fibrosis Transmembrane Conductance Regulator, as mutações mais prevalentes foram Delta F508 (45%) e S4X (18%). Os grupos de mutações apresentaram associação somente (p=0,013) com a insuficiência pancreática e não com a gravidade da doença (p=0,073). Esta última apresentou associação com a colonização por Pseudomonas aeruginosa e Staphylococcus aureus (p=0,007) e com baixo peso (p=0,036). O óbito foi associado com a idade no diagnóstico (p=0,016), a sintomatologia respiratória (p=0,013), a colonização (p=0,024), o baixo peso (p=0,017) e a ocorrência de internação (p=0,003). Conclusões: Foi possível observar associação entre as mutações e a presença de insuficiência pancreática; entre a colonização por Staphylococcus aureus e o baixo peso com a gravidade da doença; e ausência de associação entre as mutações e a gravidade da doença. Os fatores ambientais merecem ser investigados mais detalhadamente, pois parecem apresentar impacto importante na gravidade da doença.

Mechanism of Xiaoqinglong Decoction and Qingqi Huatan Pills in improving pathological airway mucus based on nuclear factor-κB/microRNA-494 signaling regulation of mucin 5AC and cystic fibrosis transmembrane conductance regulator / 中华危重病急救医学

Objective:To observe the effects of Xiaoqinglong Decoction and Qingqi Huatan Pills on interleukin-1β(IL-1β)-induced mucushypersecretion model of human airway epithelial H292 cellsand related molecules of nuclear factor-κB/microRNA-494 (NF-κB/miR-494) signaling pathway, and to explore the mechanism of the two medicines in improving pathological airway mucus.Methods:Methyl thiazolyl tetrazolium (MTT) colorimetric method was used to detect the effects of different concentrations of Xiaoqinglong Decoction and Qingqi Huatan Pills on the activity of H292 cellsinduced by IL-1β, and the appropriate concentration was selected for subsequent experiments. Cells were randomly divided into blank group, IL-1β model group (5 μg/L IL-1β), NF-κB inhibitor pyrrolidinedithiocarbamate (PDTC) group (5 μg/L IL-1β+100 μmol/L PDTC), Xiaoqinglong Decoction (5 μg/L IL-1β+1 000 mg/L Xiaoqinglong Decoction) and Qingqi Huatan Pill group (5 μg/L IL-1β+1 000 mg/L Qingqi Huatan Pills). 5 μg/L IL-1β was used to induce H292 cells for 24 hours to establish a model of airway epithelial mucus hypersecretion. Enzyme linked immunosorbent assay (ELISA) method was used to detect the levels of mucin 5AC (MUC5AC), tumor necrosis factor-α (TNF-α) and IL-8 and the synthesis of intracellular MUC5AC and cystic fibrosis transmembrane conductance regulator (CFTR). Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression levels of MUC5AC mRNA, CFTR mRNA, miR-494. Western blotting was used to detect protein expression of key proteins (p65) and NF-κB inhibitors (IκB) in NF-κB signaling pathway.Results:Xiaoqinglong Decoction and Qingqi Huatan Pills with the concentration of 1 000 mg/L were selected for the follow-up experiment. Compared with the blank group, the levels of MUC5AC, TNF-α and IL-8 were significantly increased in the model group, intracellular MUC5AC protein content and mRNA expression were also significantly increased, intracellular CFTR protein content and mRNA expression were significantly decreased, and intracellular p65 protein expression was significantly up-regulated, the expression of IκB protein was significantly down-regulated, and the expression of miR-494 was significantly increased. Compared with the model group, the levels of MUC5AC, TNF-α and IL-8 were significantly reduced in PDTC group, Xiaoqinglong Decoction group and Qingqi Huatan Pill group, intracellular MUC5AC protein content and mRNA expression were also significantly decreased, and intracellular p65 protein expression was significantly down-regulated, and IκB protein expression was significantly up-regulated, miR-494 expression was significantly reduced. Intracellular CFTR protein content and mRNA expression were significantly increased in both PDTC group and Qingqi Huatan Pill group. Compared with the PDTC group, the level of TNF-α in the Xiaoqinglong Decoction group was significantly increased (ng/L: 22.77±3.14 vs. 11.09±3.37, P < 0.05), the content and mRNA expression of CFTR and IκB protein expression was significantly decreased [CFTR protein (ng/L): 97.38±6.62 vs. 227.04±19.48, CFTR mRNA (2 -ΔΔCt): 0.99±0.08 vs. 1.21±0.08, IκB/β-actin: 1.69±0.11 vs. 2.00±0.18, all P < 0.05], the level of TNF-α in Qingqi Huatan Pill group was significantly higher (ng/L: 19.08±3.71 vs. 11.09±3.37, P < 0.05). Compared with Xiaoqinglong Decoction group, the protein content and mRNA expression of CFTR and IκB protein expression in Qingqi Huatan Pill group were significantly increased [CFTR protein (ng/L) : 235.01±22.71 vs. 97.38±6.62, CFTR mRNA(2 -ΔΔCt): 1.32±0.15 vs. 0.99±0.08, IκB/β-actin: 1.94±0.16 vs. 1.69±0.11, all P < 0.05]. Conclusions:The effect of Xiaoqinglong Decoctionin improving the hypersecretion of mucus in the airway epithelium may be related to the inhibition of NF-κB/miR-494 inflammatory signal-mediated MUC5AC hypersecretion, while the effect of Qingqi Huatan Pills may be related to the inhibition of NF-κB/miR-494 inflammatory signal-mediated MUC5AC hypersecretion and CFTR dysfunction. Therefore, the difference in the mechanism of the two treatments of airway pathological mucus is mainly in the regulation of CFTR mRNA and protein.

Research progress of cystic fibrosis associated Pseudo-Bartter syndrome / 中华实用儿科临床杂志

Cystic fibrosis (CF) is an autosomal recessive inherited disease affecting multiple body systems.Pseudo-Bartter syndrome (PBS) is a common manifestation of CF, with such clinical features as hypochloremia, hyponatremia, hypokalemia and metabolic alkalosis.However, PBS patients do not have renal tubulopathy.Children with CF are prone to develop electrolyte abnormalities due to fluid and electrolyte loss.In this article, the pathogenesis, clinical manifestations, diagnosis, and treatment of CF associated PBS were reviewed in order to enhance clinical understan-ding of this disease.

Advances in the pathogenesis and molecular regulatory therapy of cystic fibrosis / 中华实用儿科临床杂志

Cystic fibrosis (CF) is an autosomal recessive monogenetic disease caused by homozygous and double-heterozygous mutations of the cystic fibrosis transmembrane conductance regulator ( CFTR)gene.The CFTR gene encodes a chloride ion channel protein, which is distributed in the airway, pancreatic duct epithelium and other body parts.CFTR dysfunction will give rise to a series of clinical manifestations.Remarkable achievements have been made in specific molecular regulatory therapy targeting CFTR dysfunction over the past 10 years.CF has thus become a model disease, which provides new approaches for the research on genetics, molecular and cellular pathogenesis, and drug discovery of other rare genetic diseases.In this article, the latest research progress on the pathogenesis and molecular regulatory therapy of CF in recent years was reviewed to improve clinical understanding of the disease mechanism and molecular regulatory therapy.

Research progress of cystic fibrosis and its therapies / 药学学报

Cystic fibrosis (CF) is a common and life-threatening autosomal recessive disorder in Caucasians populations. The disease is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. The absence of mature CFTR at the correct cellular location or dysfunction of CFTR proteins has been observed in CF patients. CF is frequently accompanied by a variety of complex complications, such as impairments in pulmonary functions, which may lead to successive infections and respiratory failure. Recently, with the understanding of the pathogenesis of CF, a wide array of therapeutic strategies for the treatment of CF has been designed. This review summarizes pathogenic mechanisms of CF, mechanisms of action of drugs, routes of administration, and new drug development as well as provides insights into the advanced treatment strategies for CF.

Establishment of a CFTR-based detection method for the second messenger cAMP in the cytoplasm / 中国应用生理学杂志

Objective: To establish a detection method based on Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) that can sensitively detect the second messenger cyclic AMP (cAMP) in the cytoplasm. Methods: The eukaryotic expression vectors of CFTR and YFP-H148Q / I152L were constructed respectively. FRT cells co-expressing CFTR and YFP-H148Q / I152L were obtained by liposome transfection. The expression of CFTR and YFP-H148Q / I152L in FRT cells was observed by an inverted fluorescence microscopy, and flow cytometry was used to detect the purity of cells; The cell model was identified by the fluorescence quenching kinetics test. The validation of the cell model which could screen CFTR modulators was verified by the fluorescence quenching kinetics experiments. The radioimmunoassay was used to detect the cAMP concentration in cytoplasm after adding CFTR activator. Results: The results of the inverted fluorescence microscope showed that CFTR was expressed in the cell membrane and YFP-H148Q / I152L was expressed in the cytoplasm of FRT cells. The FRT cell model stably co-expressing ANO1 and YFP-H148Q / I152L was successfully constructed. The model could screen CFTR modulators, and the slope of fluorescence change and the concentration of CFTR modulators were in a dose-dependent manner. The slope of the fluorescence could reflect the cAMP concentration in the cytoplasm. The cell model could sensitively detect the intracellular cAMP concentration. Conclusion: The cell model could efficiently and sensitively detect the second messenger cAMP concentration in the cytoplasm, and it provided a simple and efficient method for the study of other targets associated cAMP signal.

Respiratory syncytial virus persistent infection causes acquired CFTR dysfunction in human bronchial epithelial cells / 中南大学学报(医学版)

Objective: Many studies have shown that respiratory syncytial virus persistent infection may be the main cause of chronic respiratory pathology. However, the mechanism is unclear. Cystic fibrosis transmembrane conduction regulator (CFTR) is an apical membrane chloride channel, which is very important for the regulation of epithelial fluid, chloride ion, and bicarbonate transport. CFTR dysfunction will lead to changes in bronchial secretions and impair mucus clearance, which is related to airway inflammation. In our previous study, we observed the down-regulation of CFTR in airway epithelial cells in respiratory syncytial virus (RSV) infected mouse model. In this study, we further investigated the expression and function of CFTR by constructing an airway epithelial cell model of RSV persistent infection. Methods: 16HBE14o- cells were infected with RSV at 0.01 multiplicity of infection (MOI). The expression of CFTR was detected by real-time RT-PCR, immunofluorescence, and Western blotting. The intracellular chloride concentration was measured by N-(ethoxycarbonylmethyl)-6-methoxyquinolium bromide (MQAE) and the chloride current was measured by whole-cell patch clamp recording. Results:16HBE14o-cells infected with RSV were survived to successive passages of the third generation (G3), while the expression and function of CFTR was progressively decreased upon RSV infection from the first generation (G1) to G3. Exposure of 16HBE14o-cells to RSV led to the gradual increase of TGF-β1 as well as phosphorylation of Smad2 following progressive RSV infection. Disruption of TGF-β1 signaling by SB431542 prevented Smad2 phosphorylation and rescued the expression of CFTR. Conclusion:RSV infection can lead to defective CFTR function in airway epithelial cells, which may be mediated via activation of TGF-β1 signaling pathway.

Multi-disciplinary team of the treatment of bilateral lung transplantation for pediatric cystic fibrosis / 器官移植

Objective To improve the surgical efficacy of bilateral lung transplantation for pediatric cystic fibrosis through multi-disciplinary team (MDT). Methods Preoperative MDT consultation was delivered for a 10-year-old child with rare end-stage cystic fibrosis to establish the corresponding treatment protocol. Results The child was diagnosed with cystic fibrosis for 5 years, and the indication of lung transplantation were confirmed. After preoperative MDT consultation, bilateral lung transplantation via a Clam-shell incision was determined. The vital signs were maintained stable during operation. Postoperatively, ventilator-assisted ventilation, anti-infection, immunosuppression, acid suppression, prevention of stress ulceration and other treatments were delivered. Individualized treatment was given according to the characteristics of the child, and the child was well recovered. Conclusions Through preoperative MDT consultation, lung transplantation yields satisfactory surgical efficacy in treating pediatric cystic fibrosis and lowers the risk of postoperative complications, which deserves application in clinical practice.

The role of biochemical testing in cystic fibrosis

@#Cystic fibrosis (CF) is a life-limiting autosomal recessive disorder as a result of CF transmembrane conductance regulator gene mutation. It has a wide range of disease severity in patients with the same genotype. Case report: A 5-year-old Malay boy with a history of recurrent pneumonia, presented with productive cough, fever and worsening tachypnoea. Physical examination revealed coarse crepitations, reduced breath sounds and clubbing. Biochemical investigations showed that he had respiratory type 2 failure as a result of bronchiectasis. Sweat conductivity done twice was raised supporting a diagnosis of CF. Other investigations such as bronchoscopy to look for congenital anomaly of the lung, infectious disease screening and tuberculosis, fungal and viral culture and sensitivity were negative. Further cascade screening revealed high sweat conductivity results in his siblings. Discussion: Although CF prevalence is low in Malaysia, it is nevertheless an important diagnosis to be recognised as it is associated with increased morbidity.

Efficacy of Qingfei Huatantang in Treatment of COPD by Up-regulating Expression of CFTR Through JAK2/STAT3 Pathway / 中国实验方剂学杂志

Objective:To explore the efficacy and mechanism of Qingfei Huatan Tang on chronic obstructive pulmonary disease (COPD). Method:The rat model of COPD was established through smoke inhalation combined with lipopolysaccharide (LPS) and pulmonary compound injection. After successful modeling, the rats were randomly divided into 6 groups, namely the control group, the COPD model group, low, medium and high-dose Qingfei Huatan Tang groups and the ambroxol group. After 28 days of modeling, the drug was administered. Low, medium and high-dose Qingfei Huatan Tang (7.5, 15, 30 g·kg-1) and ambroxol (35 mg·kg-1) were administered continuously for 14 days. Immunohistochemistry and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect protein expression and mRNA expression of cystic fibrosis transmembrane conductance regulator (CFTR) in pulmonary fibrosis. NCI-H292 cells were induced by LPS to establish a mucus hypersecretion model. The experiment was divided into 8 groups, namely the blank control group, LPS group, LPS+10% fetal bovine serum group, LPS+ physiological serum group, LPS+5% drug serum group, LPS+10% drug serum group, LPS+20% drug serum group and LPS+AG490 group. Immunofluorescence, Western blot and Real-time PCR were used to observe the protein and mRNA expressions of CFTR in NCI-H292 cells after LPS stimulation, and western blot was used to detect the expression of tyrosine kinase 2/transcription factor 3 (JAK2/STAT3) signaling pathway in NCI-H292 cells after LPS stimulation. Result:There were a large number of brown particles around the lumen of lung tissues in the normal group, with increased COPD expression. There were a few brown particles around the lumen of lung tissues in the model group compared with the normal group, with decreased COPD expression. Compared with the normal group, mRNA and protein expressions of CFTR in the lung tissues of the COPD model group were significantly decreased (P<0.05). Compared with the model group, mRNA and protein expressions of CFTR in the lung tissues of low, medium and high-dose Qingfei Huatan Tang groups (P<0.05). Compared with the blank control group, mRNA and protein expressions of CFTR in NCI-H292 cells of the LPS group (P<0.05), with significant increases in protein expressions of p-JAK2 and p-STAT3 (P<0.05). Compared with the model group, 5%, 10%, 20% Qingfei Huatan Tang-containing serum groups showed significant increases in mRNA and protein expressions of CFTR, but with significant decreases in p-JAK2, p-STAT3 protein expressions (P<0.05, P<0.01). Conclusion:Qingfei Huatan Tang up-regulated CFTR in the treatment of COPD by inhibiting JAK2/STAT3 pathway.

Cystic fibrosis lung disease: Current perspectives

Cystic fibrosis (CF) is an autosomal recessive disease caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR). These mutations alter the synthesis, processing, function, or half-life of CFTR, the main chloride channel expressed in the apical membrane of epithelial cells in the airway, intestine, pancreas, and reproductive tract. Lung disease is the most critical manifestation of CF. It is characterized by airway obstruction, infection, and inflammation that lead to fatal tissue destruction, which causes most CF morbidity and mortality. This article reviews the pathophysiology of CF, recent animal models, and current treatment of CF.

Lung transplantation for cystic fibrosis: a case report and literature review / 器官移植

Objective To investigate the clinical efficacy and prognosis of lung transplantation in the treatment of cystic fibrosis (CF). Methods Clinical data of one patient with end-stage CF undergoing allogeneic bilateral lung transplantation were retrospectively analyzed. Clinical characteristics, diagnostic methods and treatment strategies of the CF recipient were summarized. Results The recipient had suffered from relevant symptoms since childhood including repeated cough and purulent sputum for 30 years, complicated with recurrent pulmonary infection combined with acute exacerbation, chronic sinusitis and extremely severe malnutrition. Prior to lung transplantation, the patient had to depend upon the invasive ventilator due to respiratory muscle weakness, and admitted to intensive care unit (ICU) for a long time. Imaging examination revealed multiple cystic columnar bronchiectasis accompanied with infection in bilateral lungs. The diagnosis of CF was further confirmed by sweat test and gene detection. The recipient underwent bilateral lung transplantation on August 17, 2017 and received rehabilitation treatment. The lung function was gradually restored to normal. The recipient had obtained the same quality of life to the healthy counterparts since the date of manuscript submission (over 2 years). Conclusions Lung transplantation is an efficacious treatment for end-stage CF, which can not only save patients' lives, but also significantly improve the quality of life of patients.

Moduladores CFTR (cystic fibrosis transmembrane conductance regulator): presente y futuro en la terapia de fibrosis quística. Revisión / Cystic fibrosis transmembrane conductance regulator modulators: Present and future in cystic fibrosis treatment. A review

Los moduladores cystic fibrosis transmembrane conductance regulator (CFTR) representan el presente y el futuro del manejo farmacológico para los pacientes con fibrosis quística. El objetivo de esta publicación es realizar una revisión de esta opción terapéutica. Se revisaron artículos científicos consultando las bases de datos MedLine, información disponible a través de la página oficial Cystic Fibrosis Foundation, desde 2009 hasta 2018, en el idioma inglés. Sin restricciones respecto al tipo de estudio, se seleccionaron 12 artículos que incluyeron información sobre el estado actual de la investigación sobre moduladores CFTR. Actualmente, están aprobados por la Food and Drug Administration tres moduladores: ivacaftor, lumacaftor + ivacaftor y tezacaftor + ivacaftor, y hay otros 11 en diferentes fases de estudio. La terapia con moduladores CFTR es una realidad en desarrollo que apunta al máximo objetivo de la medicina personalizada y que promete mejorar la calidad de vida de pacientes con fibrosis quística.

Cystic fibrosis transmembrane conductance regulator (CFTR) modulators are the present and future of drug management for patients with cystic fibrosis. The objective of this article is to review this therapeutic option. Scientific articles were reviewed by searching the MedLine database, which is available through the Cystic Fibrosis Foundation's official website, from 2009 to 2018, in English. Twelve articles about the current status of research in CFTR modulators were selected without restrictions regarding the type of study. To date, the United States Food and Drug Administration has approved three modulators: ivacaftor, lumacaftor + ivacaftor, and tezacaftor + ivacaftor, while other 11 drugs are being studied in different investigation phases. CFTR modulator therapy is a developing reality aimed at the highest goal of personalized medicine and promises to improve the quality of life of cystic fibrosis patients.

Effects of estrogen on bicarbonate secretion of duodenal mucosa / 中华消化杂志

Objective@#To study the effects of estrogen on bicarbonate secretion of duodenal mucosal, and to observe estrogen receptor (ER) subtypes of estrogen.@*Methods@#Sixteen 4-week-old male C57 mice were divided into control group and estrogen group, with eight mice in each group. The mice serum level of estrogen was detected by chemiluminescence. The expression of cystic fibrosis transmembrane conductance regulator (CFTR), solute carrier family 26 (SLC26) A3 and SLC26A6 in the duodenum tissues were determined by real-time polymerase chain reaction (RT-PCR). After SCBN cells treated with estrogen, ERα and ERβ blocking agent, and transfected with silenced ERα and ERβ for 24 and 48 hours, the expression levels of CFTR, SLC26A3 and SLC26A6 mRNA in cells were detected by RT-PCR. The effects of estrogen before and after silenced ERα and ERβ on bicarbonate secretion of SCBN cells were observed by high-speed ion imaging system. T test and rank sum test were used for statistical analysis.@*Results@#Compared with that of control group, the serum estrogen level of estrogen group was significantly high ((4 874±942) pmol/L vs. (657±187) pmol/L, t=-11.579, P<0.01). The expression levels of CFTR, SLC26A3 and SLC26A6 mRNAs in duodenum tissues of estrogen group were higher than those of control group (0.856±0.302 vs. 0.452±0.246, 2.910±1.680 vs. 1.120±0.540, 1.272±0.667 vs. 0.319±0.114), and the differences were statistically significant (t=-2.317, -2.483 and -3.721, all P<0.05). Compared with those treated with estrogen for 24 and 48 hours, the levels of CFTR mRNA and SLC26A6 mRNA were lower after the ERβ blocking agent were added into estrogen for 24 and 48 hours (CFTR mRNA: 0.171±0.059 vs. 0.522±0.260 and 0.111±0.014 vs. 0.578±0.297; SLC26A6 mRNA: 0.486±0.289 vs. 1.118±0.571 and 0.492±0.231 vs. 1.551±0.605), and the differences were statistically significant (tCFTR mRNA=2.974 and 2.655, tSLC26A6 mRNA=2.393 and 3.272; all P<0.05). Compared with those of silenced ERα group, the levels of CFTR mRNA, SLC26A3 mRNA and SLC26A6 mRNA were higher after ERα silenced and then added estrogen for 24 and 48 hours (24 h: 5.073±2.270 vs. 1.185±0.494, 1.796±1.168 vs. 0.468±0.108 and 3.085±1.357 vs. 0.706±0.347; 48 h: 5.025±1.998 vs. 1.185±0.494, 1.557±0.653 vs. 0.468±0.108 and 3.290±1.750 vs. 0.706±0.347), and the differences were statistically significant (t24 h=-4.122, -2.773 and -4.162, t48 h=-4.604, -4.034 and -3.250; all P<0.05). Compared with that of silenced ERα group, the bicarbonate secretion increased after ERα silenced and then added estrogen for 24 and 48 hours (0.72±0.17 and 1.15±0.25 vs. 0.35±0.17), and pH also elevated, and the differences were statistically significant (t=-6.516 and -12.387, both P<0.01).@*Conclusion@#Estrogen mainly up-regulates the expression of bicarbonate transporter protein in duodenal mucosal epithelial cells through ERβ, and promotes the bicarbonate secretion of duodenal mucosa.