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Three-dimensional in vitro models of neuromuscular tissue

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Neural Regeneration research 2022年第4期17卷 759-766页

作者： Paolo Raffa Maria Easler Anna Urciuolo institute of pediatric research IRP PadovaItaly Molecular Medicine Department University of PadovaPadovaItaly

Skeletal muscle is a dynamic tissue in which homeostasis and function are guaranteed by a very defined three-dimensional organization of myofibers in respect to other nonmuscular components,including the extracellular matrix and the nervous network.In particular,communication between myofibers and the nervous system is essential for the overall correct development and function of the skeletal muscle.A wide range of chronic,acute and genetic-based human pathologies that lead to the alteration of muscle function are associated with modified preservation of the fine interaction between motor neurons and myofibers at the neuromuscular junction.Recent advancements in the development of in vitro models for human skeletal muscle have shown that three-dimensionality and integration of multiple cell types are both key parameters required to unveil pathophysiological relevant phenotypes.Here,we describe recent achievement reached in skeletal muscle modeling which used biomaterials for the generation of three-dimensional constructs of myotubes integrated with motor neurons.

关键词： 3D organization bioengineering biomaterials motor neurons neuromuscular junction skeletal muscle models

Upregulation of cadherin-11 contributes to cholestatic liver fibrosis

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pediatric Investigation 2022年第2期6卷 100-110页

作者： Bo Wu Xinbei Tian Weipeng Wang Jing Zhu Ying Lu Jun Du Yongtao Xiao Department of pediatric Surgery Xin Hua HospitalSchool of MedicineShanghai Jiao Tong UniversityShanghaiChina Department of pediatric Gastroenterology and Nutrition Shanghai Institute of Pediatric ResearchShanghaiChina Shanghai Key Laboratory of pediatric Gastroenterology and Nutrition ShanghaiChina

Importance:Cadherin-11(CDH11),a cell-to-cell adhesion molecule,is implicated in the fibrotic process of several organs.Biliary atresia(BA)is a common cholestatic liver disease featuring cholestasis and progressive liver fibrosis in children.Cholestatic liver fibrosis may progress to liver cirrhosis and lacks effective therapeutic strategies.Currently,the role of CDH11 in cholestatic liver fibrosis remains unclear.Objective:This study aimed to explore the functions of CDH11 in cholestatic liver fibrosis.Methods:The expression ofCDH11 in BA livers was evaluated by database analysis and immunostaining.Seven BA liver samples were used for immunostaining.The wild type(Wt)andCDH11 knockout(CDH11^(-/-))mice were subjected to bile duct ligation(BDL)to induce cholestatic liver fibrosis.The serum biochemical analysis,liver histology,and western blotting were used to assess the extent of liver injury and fibrosis as well as activation of transforming growth factor-β(TGF-β)/Smad pathway.The effect of CDH11 on the activation of hepatic stellate cell line LX-2 cells was investigated.Results:Analysis of public RNA-seq datasets showed thatCDH11 expression levels were significantly increased in livers of BA,and CDH11 was correlated with liver fibrosis in BA.BDL-induced liver injury and liver fibrosis were attenuated inCDH11^(-/-)mice compared to Wt mice.The protein expression levels of phosphorylated Smad2/3 were decreased in livers ofCDH11^(-/-)BDL mice compared to Wt BDL mice.CDH11 knockdown inhibited the activation of LX-2 cells.Interpretation:CDH11 plays an important role in cholestatic liver fibrosis and may represent a potential therapeutic target for cholestatic liver disease,such as BA.

关键词： Cadherin-11 Biliary atresia Cholestatic liver fibrosis TGF-β/Smad

Gastrointestinal viral shedding in children with SARS‑CoV‑2:a systematic review and meta‑analysis

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World Journal of pediatrics 2022年第9期18卷 582-588页

作者： Sepideh Benvari Shima Mahmoudi Mohsen Mohammadi Department of Microbiology Faculty of MedicineQazvin University of Medical SciencesQazvinIran pediatric Infectious Disease research Center Tehran University of Medical SciencesTehranIran Non-Communicable pediatric Diseases research Center Health Research InstituteBabol University of Medical SciencesBabolIran

Background Coronavirus disease 2019(COVID-19)has diferent manifestations in pediatric cases.It is assumed that they might present more gastrointestinal symptoms with a diferent viral shedding pattern in gastrointestinal samples.In this systematic review and meta-analysis,we aimed to evaluate the viral shedding pattern in gastrointestinal specimens of children with COVID-19.Methods We searched all published studies in English language in PubMed,Web of Science,and Scopus,up to date as of October 2021.Our search included the term"severe acute respiratory syndrome coronavirus 2,COVID-19,SARS-CoV-2,novel coronavirus,or coronavirus;and shed,excrete,secret,or carriage;and stool or rectal;and children or pediatrics".We included studies evaluating SARS-CoV-2 shedding in gastrointestinal specimens,including rectal swabs and stool samples of children with COVID-19 infection.We excluded duplicated data,case reports,and studies without original data.Results Twelve studies met the eligibility criteria for the qualitative synthesis,10 of which were included in the meta-analysis.The pooled prevalence of gastrointestinal severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)RNA in children with COVID-19 was 86%(95%confdence interval 73%–96%,I 2=62.28%).After respiratory specimen had become negative,72%(43/60)had persistent shedding in gastrointestinal specimens.The gastrointestinal RNA had a positive test result for more than 70 days after symptoms onset.Conclusions Gastrointestinal shedding of SARS-CoV-2 might occur in a substantial portion of children and might persist long after negative respiratory testing.Further research is recommended to fnd the role of SARS-CoV-2 gastrointestinal shedding in transmission in children.

关键词： Coronavirus disease 2019(COVID-19) Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) Viral shedding

Emerging role of regulated cell death in intestinal failure-associated liver disease

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Hepatobiliary & Pancreatic Diseases International 2024年第3期23卷 228-233页

作者： Si-Yang Cheng Lu Jiang Ying Wang Wei Cai Department of pediatric Surgery Xinhua HospitalSchool of MedicineShanghai Jiao Tong UniversityShanghai 200092China Shanghai Key Laboratory of pediatric Gastroenterology and Nutrition Shanghai 200092China Shanghai institute for pediatric research Shanghai 200092China Division of pediatric Gastroenterology and Nutrition Xinhua HospitalSchool of MedicineShanghai Jiao Tong UniversityShanghai 200092China

Intestinal failure-associated liver disease(IFALD)is a common complication of long-term parenteral nutrition that is associated with significant morbidity and mortality.It is mainly characterized by cholestasis in children and steatohepatitis in adults.Unfortunately,there is no effective approach to prevent or reverse the disease.Regulated cell death(RCD)represents a fundamental biological paradigm that determines the outcome of a variety of liver diseases.Nowadays cell death is reclassified into several types,based on the mechanisms and morphological phenotypes.Emerging evidence has linked different modes of RCD,such as apoptosis,necroptosis,ferroptosis,and pyroptosis to the pathogenesis of liver diseases.Recent studies have shown that different modes of RCD are present in animal models and patients with IFALD.Understanding the pathogenic roles of cell death may help uncover the underlying mechanisms and develop novel therapeutic strategies in IFALD.In this review,we discuss the current knowledge on how RCD may link to the pathogenesis of IFALD.We highlight examples of cell death-targeted interventions aiming to attenuate the disease,and provide perspectives for future basic and translational research in the field.

关键词： Parenteral nutrition Apoptosis Necroptosis Cholestasis Steatosis

Biliary atresia: Where do we stand now?

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World Journal of Hepatology 2016年第36期8卷 1593-1601页

作者： Krishna Kumar Govindarajan Department of pediatric Surgery Jawaharlal Institute of Postgraduate Medical Education and Research

The pathway from clinical suspicion to establishing the diagnosis of biliary atresia in a child with jaundice is a daunting task. However, investigations available help to point towards the correct diagnosis in reasonable time frame. Imaging by Sonography has identified several parameters which can be of utility in the diagnostic work up. Comparison of Sonography with imaging by Nuclear medicine can bring out the significant differences and also help in appropriate imaging. The battery of Biochemical tests, available currently, enable better understanding of the line-up of investigations in a given child with neonatal cholestasis. Management protocols enable standardized care with optimal outcome. The place of surgical management in biliary atresia is undisputed, although Kasai procedure and primary liver transplantation have been pitted against each other. This article functions as a platform to bring forth the various dimensions of biliary atresia.

关键词：胆汁的闭锁新生的 cholestasis Kasai 过程新生的黄疸 Hyperbilirubinemia

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World Journal of pediatrics 2022年第4期18卷 271-277页

作者： Yi Cao Wei-Hui Yan Li-Na Lu Yi-Jing Tao Hai-Xia Feng Qing-Qing Wu Yi-Jing Chu Wei Cai Ying Wang Division of pediatric Gastroenterology and Nutrition Xinhua Hospital Affiliated To Shanghai Jiao Tong University School of MedicineNo.1665Kongjiang Rd.ShanghaiChina Shanghai institute for pediatric research ShanghaiChina Shanghai Key Laboratory of pediatric Gastroenterology and Nutrition No.1665Kongjiang Rd.ShanghaiChina

Background Children with intestinal failure(IF)have frequent catheter-related bloodstream infections(CRBSIs).This study aimed to analyze the clinical presentation and laboratory parameters of CRBSIs in children with IF.Methods This 6-year retrospective study was conducted among IF children with CRBSIs at an intestinal rehabilitation center in China.Clinical data were collected,including data of temperature and gastrointestinal symptoms.Blood/catheter culture,fecal tests,and calculation of inflammatory index were performed,which were obtained within 1 week since CRBSI onset.Results Fifty children with 87 CRBSIs were identified,of which there were 17 suspected and 70 confirmed cases.Seventy-two pathogens were cultured from 70 positive blood cultures:63%were Gram-positive organisms,23%were Gram-negative organisms,and 11%were fungal organisms.Overall,48.6%were enteric organisms;47.2%of bacterial pathogens were consistent between fecal and blood cultures.Moreover,46.3%fecal routines showed abnormalities including increased white blood cells,occult blood positive and the presence of fat droplets.The consistent symptom at onset of CRBSIs was fever and gastrointestinal symptoms including increased stool output,abdominal distension,or both.C-reactive protein(CRP)and procalcitonin(PCT)were elevated,i.e.,16.5 mg/L[interquartile range(IQR)8.7-44.7]and 0.48 ng/mL(IQR 0.2-1.76),respectively.Conclusions IF children had a high rate of CRBSIs,of which larger proportions were due to Gram-positive and enteric organisms.Fever and/or gastrointestinal symptoms,combined with elevated CRP and PCT,is conducive to the early diagnosis of CRBSIs in IF patients.

关键词： Enteric pathogens Fecal examination Gastrointestinal symptoms

Neonatal methicillin-resistant Staphylococcus aureus pneumonia-related recurrent fatal pyopneumothorax: A case report and review of literature

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World Journal of Clinical Cases 2023年第30期11卷 7475-7484页

作者： Xing-Chao Li Li Sun Tao Li Department of pediatrics Taihe HospitalHubei University of MedicineShiyan 442000Hubei ProvinceChina institute of pediatric research Hubei University of MedicineShiyan 442000Hubei ProvinceChina institute of pediatric research Taihe HospitalHubei University of MedicineShiyan 442000Hubei ProvinceChina

BACKGROUND Although neonatal Staphylococcus aureus pneumonia is common and usually curable,it can also be refractory and life-threatening.Herein,we report a case of severe neonatal community-acquired methicillin-resistant Staphylococcus aureus(CA-MRSA)necrotizing pneumonia with bilateral recurrent pyopneumothorax,respiratory failure,heart failure,and cardiac arrest.We hope our report will add to the understanding of this disease.CASE SUMMARY An 18-d-old boy presented with cough for five days,fever for three days,and dyspnea for two days.Preadmission chest radiograph revealed high-density shadows in both lungs.On admission,his oxygen saturation fluctuated around 90%under synchronized intermittent mandatory ventilation.He was unconscious,with dyspnea,weak heart sounds and hepatomegaly.Moist crackles were present throughout his left lung,while the breath sounds in the right lung were decreased.After high-frequency oscillatory ventilation,empiric antimicrobials(meropenem and vancomycin),improved circulation,and right pleural cavity drainage for right pneumothorax(approximately 90%compression),his oxygen saturation level stayed above 95%,and recruitment of the right lung was observed.His condition did not deteriorate until the 5th day of hospitalization(DOH 5).On the morning of DOH 5,his oxygen saturation decreased.Subsequent chest radiograph showed bilateral pneumothorax with nearly 100%compression of the left lung.Desaturation was not relieved after urgent left pleural cavity drainage,and cardiac arrest occurred soon thereafter.Although his spontaneous heartbeat returned through emergency resuscitation and salvage antibacterial therapy(linezolid and levofloxacin)was administered given the detection and antimicrobial susceptibility of MRSA,he showed no improvement,with recurrent pyopneumothorax and continued drainage of purulent fluid and necrotic lung tissue fragments from the pleural cavity.Eventually,his parents refused extracorporeal membrane oxygenation(ECMO)and gave up all the treatments,and the newborn passed

关键词： Newborn Methicillin-resistant Staphylococcus aureus Pyopneumothorax Linezolid Extracorporeal membrane oxygenation Case report

Evaluation of direct intramural injection to the bladder wall as a method for developing orthotopic tumor models

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Animal Models and Experimental Medicine 2022年第6期5卷 575-581页

作者： Masoud Bitaraf Samad Muhammadnejad Ashkan Azimzadeh Saman Behboodi Tanourlouee Erfan Amini Masoumeh Majidi Zolbin Abdol-Mohammad Kajbafzadeh pediatric Urology and Regenerative Medicine research Center GeneCell and Tissue Research InstituteTehran University of Medical SciencesTehranIran Gene Therapy research Center Digestive Diseases Research InstituteTehran University of Medical SciencesTehranIran Uro-oncology research Center Tehran University of Medical SciencesTehranIran

Background:Bladder cancer poses a great burden on society and its high rate of recurrence and treatment failure necessitates use of appropriate animal models to study its pathogenesis and test novel treatments.Orthotopic models are superior to other types since they provide a normal microenvironment.Four methods are described for developing bladder cancer models inside the animal’s bladder.Direct intramural injection is one of these methods and is widely used.However,its efficacy in model development has not yet been studied.We aimed to evaluate the efficacy and success rate of the direct intramural injection method of developing an orthotopic model for the study of bladder cancer.Method:Tumor cell lines were prepared in four microtubes.Aliquots of 200×10^(3) cells were injected through a 27 gauge needle into the ventral wall of the bladders of 4male and 4 female BALB/c mice following a midline 1 cm laparotomy incision.In addition,1 million cells from each microtube were injected into the flanks of control mice.To prevent infection and alleviate pain,5 mg/kg enrofloxacin and 2.5 mg/kg flunixin meglumine,respectively,were injected subcutaneously.Results:Tumors formed in all mice,resulting in 100% take rate and zero post-operation mortality.Surgery time was≤15 min per mouse.In two mice,tumors were found in the peritoneal space as well.Conclusion:Direct intramural injection is a rapid,reliable,and reproducible method for developing orthotopic models of bladder cancer.It can be done on both male and female mice and only requires readily available surgical tools.However,needle track can result in cell spillage and peritoneal tumors.

关键词： animal model bladder cancer bladder wall injection model evaluation orthotopic model

Intranasal nerve growth factor for prevention and recovery of the outcomes of traumatic brain injury

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Neural Regeneration research 2023年第4期18卷 773-778页

作者： Luigi Manni Giorgio Conti Antonio Chiaretti Marzia Soligo institute of Translational Pharmacology National Research Council of Italy(CNR)RomeItaly Intensive pediatric Therapy and pediatric Trauma Center Department of EmergencyAnesthesiological and Reanimation SciencesFondazione Policlinico Universitario A.Gemelli IRCCSRomeItaly institute of pediatrics Department of Woman and Child HealthFondazione Policlinico Universitario A.Gemelli IRCCSRomeItaly

Traumatic brain injury is one of the main causes of mortality and disability worldwide.Traumatic brain injury is characterized by a primary injury directly induced by the impact,which progresses into a secondary injury that leads to cellular and metabolic damages,starting in the first few hours and days after primary mechanical injury.To date,traumatic brain injury is not targetable by therapies aimed at preventing and/or limiting the outcomes of secondary damage but only by palliative therapies.Nerve growth factor is a neurotrophin targeting neuronal and non-neuronal cells,potentially useful in preventing/limiting the outcomes of secondary damage in traumatic brain injury.This potential has further increased in the last two decades since the possibility of reaching neurotrophin targets in the brain through its intranasal delivery has been exploited.Indeed,molecules intranasally delivered to the brain parenchyma may easily bypass the blood-brain barrier and reach their therapeutic targets in the brain,with favorable kinetics,dynamics,and safety profile.In the first part of this review,we aimed to report the traumatic brain injury-induced dysfunctional mechanisms that may benefit from nerve growth factor treatment.In the second part,we then exposed the experimental evidence relating to the action of nerve growth factor(both in vitro and in vivo,after administration routes other than intranasal)on some of these mechanisms.In the last part of the work,we,therefore,discussed the few manuscripts that analyze the effects of treatment with nerve growth factor,intranasally delivered to the brain parenchyma,on the outcomes of traumatic brain injury.

关键词： intranasal delivery nerve growth factor pharmacology traumatic brain injury