Infección grave diseminada por adenovirus en receptores de trasplante hepático / Severe adenovirus infections in hepatic transplant recipients / Infecção grave por adenovírus disseminado em receptores de transplante de fígado

Resumen: Los adenovirus humanos continúan siendo un desafío en la práctica clínica, se trata de virus de tipo ADN, con amplia distribución en la población, causa frecuente de enfermedad autolimitada en niños. Sin embargo, en pacientes inmunosuprimidos, principalmente trasplantados de células hematopoyéticas y de órganos sólidos, puede llegar a ser causa de enfermedad diseminada severa, con elevada morbimortalidad. Compartimos el primer reporte nacional de enfermedad por adenovirus diseminado, en el que se presentan dos casos clínicos de pacientes trasplantados hepáticos, que desarrollan enfermedad diseminada grave por adenovirus. Se realiza una revisión de la literatura sobre el tema.

Abstract: Human adenoviruses still constitute a challenge in the clinical practice. These are DNA viruses that are widely disseminated among the population and often cause self-limiting diseases in children. However, it may result in a severe diseminated disease with high morbi-mortality rates in immunocompromised patients, in particular transplant recipients of hematopoietic cells and solid organs. The study shares the first national report for disseminated adenovirus disease, presenting two clinical cases of hepatic transplant recipients who developed severe disseminated adenovirus disease. And conducting a literature review on the topic.

Resumo: Os adenovírus humanos continuam sendo um desafio na prática clínica; são vírus do tipo DNA, com ampla distribuição na população, causa frequente de doença autolimitada em crianças. Entretanto, em pacientes imunossuprimidos, principalmente receptores de transplante de células hematopoiéticas e órgãos sólidos, pode se tornar causa de doença disseminada grave, com alta morbimortalidade. Este é o primeiro relato nacional de doença de adenovírus disseminado, no qual são apresentados dois casos clínicos de pacientes transplantados de fígado que desenvolvem doença de adenovírus disseminada grave. Realizou-se também revisão da literatura sobre o assunto.

Adenovirus: de la neumonía a la bronquiolitis obliterante / Adenovirus: from pneumonia to obliteran bronchiolitis

Post-infectious bronchiolitis obliterans (BOPI) is a chronic obstructive disease, resulting from an acute injury and an abnormal repair process, with diffuse pulmonary fibrosis and peribronchiolar fibrosis, which cause chronic respiratory failure with prolonged oxygen dependence. The most common cause of this disease is severe bronchiolitis / pneumonia due to adenovirus (ADV), mainly in group B, before 2 years of age. In its pathogenesis are factors of the host and the characteristics of the virus that has mechanisms to prevent immunity and cause a chronic infection with great inflammatory response. This involves numerous cells (mainly lymphocytes) and cytokines that are produced by a chronic infection by ADV, which maintains a prolonged inflammatory process, determining different degrees of lung damage. In this article we will discuss the mechanisms by which this damage occurs.

La bronquiolitis obliterante postinfecciosa (BOPI) es una enfermedad obstructiva crónica, resultante de una injuria aguda y un proceso de reparación anómalo, con fibrosis pulmonar y peribronquiolar difusa, que causan insuficiencia respiratoria crónica con dependencia de oxigeno prolongada. La causa más frecuente de esta enfermedad es una bronquiolitis/neumonía grave por adenovirus (ADV), principalmente del grupo B, antes de los 2 años de vida. En su patogenia intervienen factores del huésped y las características del virus que tiene mecanismos para evitar la inmunidad y provocar una infección crónica con gran respuesta inflamatoria. En esta participan numerosas células (principalmente linfocitos) y citoquinas que se producen por una infección crónica por ADV, lo que mantiene un proceso inflamatorio prolongado, determinando distintos grados de daño pulmonar. En este artículo abordaremos los mecanismos por los cuales se produce este daño.

Función pulmonar en bronquiolitis obliterante postinfecciosa / Pulmonary function in post-infectious bronchiolitis obliterans

Bronchiolitis obliterans is a rare and severe chronic lung disease resulting from a lower respiratory tract injury. It may occur after a bone marrow or lung transplantation, infectious diseases, or less frequently after inhaling toxic substances or connective tissue diseases. Pathogenesis and molecular biology, as well as the best treatment of bronchiolitis obliterans, remain the subject of ongoing research. This review discusses our current knowledge of lung function of post-infectious bronchiolitis obliterans.

La bronquiolitis obliterante es una enfermedad pulmonar crónica rara y grave que resulta de una lesión del tracto respiratorio inferior. Puede ocurrir después de un trasplante de médula ósea o pulmón, enfermedades infecciosas, o menos frecuentemente después de inhalar sustancias tóxicas o después de enfermedades del tejido conectivo. La patogénesis y la biología molecular, así como el mejor tratamiento de la bronquiolitis obliterante, siguen siendo objeto de investigación. Esta revisión analiza nuestro conocimiento actual sobre la función pulmonar de los pacientes con bronquiolitis obliterante secundaria a infecciones.

Fatal nosocomial hemorrhagic enterocolitis probably caused by adenovirus: report of one case / Enterocolitis hemorrágica fatal nosocomial probablemente causada por adenovirus: informe de un caso

ABSTRACT Adenovirus (ADV) is a recognized cause of severe disease among immunocompromised patients. We report a previously healthy 39-year-old female, admitted with influenza pneumonia and evolving with lung hemorrhage and acute renal failure requiring mechanical ventilation and hemodialysis. She received high corticosteroid doses due to an initial suspicion of alveolar hemorrhage. Lymphopenia already present before steroid use (567/μL), was maintained during the whole hospital stay (mean 782/μL). From the second week of admission she presented a high-volume diarrhea (mean 2.5 L/day) associated to intermittent bloody stools. An ulcerative enterocolitis was confirmed by CT images and colonoscopy. ADV was detected in a colonic tissue sample by real time PCR but not by a commercial filmarray test. Cidofovir-probenecid and racecadotril therapy were indicated without changing the clinical course of diarrhea and the patient finally died.

Adenovirus (ADV) es una causa reconocida de enfermedades graves en pacientes inmunocomprometidos. Informamos el caso de una mujer de 39 años, previamente sana, que ingresó por neumonía grave por influenza, evolucionando con hemorragia pulmonar y falla renal aguda, requiriendo ventilación mecánica y hemodiálisis. Recibió altas dosis de corticoides por la sospecha inicial de una hemorragia alveolar. Tuvo linfopenia durante toda su estadía (promedio 782/μL), la que ya estaba presente antes del uso de los corticoides (567/μL). Desde la segunda semana de hospitalización, presentó una diarrea de alto volumen (promedio 2,5 L/día) asociada a la presencia de sangre en deposiciones en forma intermitente. Se confirmó una enterocolitis ulcerativa por tomografía computada y colonoscopía. Se detectó ADV en muestras de biopsia colónica por PCR en tiempo real pero no por un test de PCR múltiples automatizado comercial. Fue tratada con cidofovir-probenecid y racecadrotrilo sin impacto clínico y la paciente finalmente falleció.

Bilateral disciform keratitis of presumed adenoviral etiology

Adenoviral conjunctivitis may lead to subepithelial corneal infiltrates as a late complication. Herein, we aim to present a 19-year-old healthy female, who developed bilateral disciform keratitis three weeks after suffering adenoviral conjunctivitis. She presented with widespread subepithelial corneal infiltrates in addition to central corneal edema with white distinct border resembling immune stromal ring, as well as Descemet's folds and keratic precipitates in the central area. Following topical corticosteroid and ganciclovir for 10 days, her condition improved. After 1 month, she had another episode. Short-term topical corticosteroids in addition to long-term topical cyclosporine and nonpreserved artificial tears were able to prevent further recurrences.

Disseminated adenovirus infection in a 10-year-old renal allograft recipient

Disseminated adenovirus infection can result in high mortality and morbidity in immunocompromised patients. Here, we report the case of a 10-year-old renal allograft recipient who presented with hematuria and dysuria. Adenovirus was isolated from his urine. His urinary symptoms decreased after intravenous hydration and reduction of immunosuppressants. However, 2 weeks later he presented with general weakness and laboratory tests indicated renal failure necessitating emergency hemodialysis. Adenovirus was detected in his sputum; therefore, intravenous ganciclovir and immunoglobulin therapy were initiated. Renal biopsy revealed diffuse necrotizing granulomatous tubulointerstitial nephritis compatible with renal involvement of the viral infection. Adenovirus was detected in his serum. Despite cidofovir administration for 2 weeks, adenovirus was also detected in the cerebrospinal fluid, resulting in generalized tonic-clonic seizure. The patient died 7 weeks after the onset of urinary symptoms. Adenovirus should be considered in screening tests for post-renal transplantation patients who present with hemorrhagic cystitis.

Comparison of Clinical Manifestation and Laboratory Findings between Adenoviral Infection with or without Kawasaki Disease / 이화의대지

OBJECTIVES: Adenovirus infection, which has been known to mimic Kawasaki disease (KD), is one of the most frequent conditions observed during differential diagnosis when considering KD. Accordingly, it is essential to being able to differentiate between these two diseases. Therefore, we performed multiplex reverse transcriptase-polymerase chain reaction and tissue-Doppler echocardiography to distinguish between adenovirus patients and KD patients. METHODS: A total of 113 adenoviral infection patients (female 48, male 65) diagnosed from January 2010 to June 2016 were evaluated. We divided adenoviral infection patients into two groups: group 1, which consisted of individuals diagnosed with KD according to the KD American Heart Association criteria (n=62, KD with adenovirus infection); and group 2, which comprised individuals only diagnosed with adenovirus infection (n=51). Laboratory data were obtained from each patient including N-terminal pro-brain natriuretic peptide. Echocardiographic measurements were compared between two groups. In addition, reverse transcriptase-polymerase chain reaction was performed using nasopharyngeal secretions to diagnose adenoviral infection. RESULTS: Conjunctival injection, cervical lymphadenopathy, polymorphous skin rash, abnormalities of the lip or oral mucosa and abnormalities of extremities were significantly higher in group 1 than group 2. Moreover, group 1 had significantly higher C-reactive protein and alanine aminotransferase levels, as well as lower platelet counts and albumin levels than group 2. Coronary artery diameter was significantly greater in group 1 than group 2. CONCLUSION: In patients with adenoviral infection with unexplained prolonged fever, echocardiography and C-reactive protein can be used to differentiate KD with adenoviral infection from adenoviral infection alone.

Adenovirus 36 y su potencial contribución en el desarrollo de obesidad / The potential contribution of adenovirus 36 to the development of obesity

The evidence of the last 20 years shows a link between viral infections and obesity in animals and humans. There are five adenovirus which have been associated with development of obesity in animals. SMAM-1 virus was the first studied in humans associated with obesity. There is compelling evidence that Ad-36 virus could contribute to the development of obesity in humans and it is related with body mass index (BMI). This manuscript reviews the association between Ad-36 and the other four virus infections with obesity. An electronic search of articles in the databases PubMed and Scielo, with use of key words: obesity, infection, adipose tissue, Ad-36, 3T3-L1 was performed. The search was restricted "human" and "animals". The importance of the relationship between virus infections and obesity has increased over the past two decades. Ad-36 shows more compelling evidence in humans. There are reports involving this virus in the enhancement of adipogenesis, adipocyte differentiation, a lower secretion of leptin and an increased insulin sensitivity. Future work should focus in larger cohort studies to confirm this association, which explains the global obesity epidemic from a new perspective.

Enfermedad de Kawasaki e infección por adenovirus en un lactante / Kawasaki disease and adenovirus infection in an infant

La enfermedad de Kawasaki es una vasculitis febril aguda propia de la infancia con afectación de vasos de pequeño y mediano calibre. Predomina en niños de entre 1 a 5 años, siendo excepcional en lactantes. El diagnóstico se basa en criterios clínicos, sin embargo debido a su frecuente presentación atípica genera dificultades diagnósticas que pueden determinar retardo en la instauración del tratamiento lo que repercute negativamente en el pronóstico. Se presenta el caso clínico de un lactante de 3 meses, previamente sano, que en el curso de una infección respiratoria por adenovirus desarrolla enfermedad de Kawasaki. La infección viral previa y/o concomitante, descrita en la literatura como un posible factor desencadenante en individuos genéticamente predispuestos, planteó mayores dificultades al equipo tratante. El objetivo de esta comunicación es alertar sobre la presentación de esta enfermedad en asociación con infecciones virales en edades precoces, para contribuir al diagnóstico y tratamiento oportunos.

Kawasaki disease (KD) is an acute febrile vasculitis of early childhood which affects small and medium blood vessels. It prevails in children between 1 to 5 years old, being it unusual in children younger than one year old. Diagnosis is based on clinical criteria, although, due to its frequently atypical presentation, it is difficult to diagnose, what may result in a delayed initiation of treatment and the subsequent negative impact on prognosis. The study presents the clinical case of a 3 month old child with a healthy medical record, who developed Kawasaki disease during a respiratory infection caused by adenovirus. A previous viral infection and/or concomitant, posed greater difficulties to the treating team because according to literature. Viral infections are described as possible triggering factors in individuals who are genetically predisposed. The objective of this comunication is to warn on how this medical condition can be associated to viral infections at early ages as well as to contribute to early diagnosis and treatment.

Recombinase polymerase amplification combined with a lateral flow dipstick for rapid and visual detection of adenovirus / 军事医学

Objective To develop a rapid, accurate, visual, and portable detection method for adenovirus types B (AdvB) and E ( AdvE).Methods Universal primers were targeted on type-specific conserved regions to allow the simultaneous detection of both human Adv (HAdV) species.A detection method based on the combination of recombinase polymerase amplification ( RPA) and lateral flow dipstick ( LFD) was established the sensitivity and specificity evaluated , and throat swab specimens of 19 patients infected with AdvB and AdvE as well as 10 healthy volunteers were detected with this method.Results The detection limit of the method was 10 copies/μl Adv DNA, which was close to that of qPCR , and there were no cross-reactions with other species of Adv and unrelated virus .The detection could be finished within 15 to 20 min within the temperature range of 25 to 45℃.When applied to clinical samples , this method showed 100% sensitivity and specificity.Conclusion This detection assay is a sensitive , specific, rapid and simple method that eliminates the need for expensive equipment , trained personnel or laboratories .The characteristics of this system render it suitable for use in grass-roots healthcare departments , and the system is especially effective for field testing and on-site testing.

Recombinase polymerase amplification combined with a lateral flow dipstick for rapid and visual detection of adenovirus / 军事医学

Objective To develop a rapid, accurate, visual, and portable detection method for adenovirus types B (AdvB) and E ( AdvE).Methods Universal primers were targeted on type-specific conserved regions to allow the simultaneous detection of both human Adv (HAdV) species.A detection method based on the combination of recombinase polymerase amplification ( RPA) and lateral flow dipstick ( LFD) was established the sensitivity and specificity evaluated , and throat swab specimens of 19 patients infected with AdvB and AdvE as well as 10 healthy volunteers were detected with this method.Results The detection limit of the method was 10 copies/μl Adv DNA, which was close to that of qPCR , and there were no cross-reactions with other species of Adv and unrelated virus .The detection could be finished within 15 to 20 min within the temperature range of 25 to 45℃.When applied to clinical samples , this method showed 100% sensitivity and specificity.Conclusion This detection assay is a sensitive , specific, rapid and simple method that eliminates the need for expensive equipment , trained personnel or laboratories .The characteristics of this system render it suitable for use in grass-roots healthcare departments , and the system is especially effective for field testing and on-site testing.

Negative immune regulatory molecule TIPE2 for treating SLE mice through regulating macrophage subtype / 重庆医学

Objective To investigate the role of tumor necrosis factor (TNF)-alpha-induced protein 8-like 2 (TIPE2) for regulating the macrophage polarization in systemic lupus erythematosus and its curative effects on experimental SLE mice.Methods The mice were treated with activated lymphocytes derived DNA (ALD-DNA) for inducing mice model,randomly divided into AAV-scr control group and AAV-TIPE2 experimental group,and injected with AAV-TIPE2 or AAV-scr virus solution from the tail vein of mice.The expression of TIPE2 mRNA and protein in polarized macrophages,serum dsDNA antibody titer,urine protein and renal pathological index were detected.Results (1) The TIPE2 expression level of TIPE2 mRNA and protein in AAV-TIPE2-transfected cells was 13.5±1.6 times and 10.8±1.6 times of AAV-scr control group respectively.(2) M2 macrophage specific molecule MGL+ was 59.6% in AAV-TIPE2 group and MGL + cells in the AAV-scr group was 8.4%.M2/M1 odds ratio of AAV-TIPE2 experimental group to AAV-scr control group was 16.(3) The recombinant TIPE2 adenovirus related vector could stably expressed in transfected HEK-293.In vitro and in vivo experiments confirmed that AAV-TIPE2 was able to induce M2 polarization of macrophages in ALD-DNA-induced lupus mice.(4) The serum anti-dsDNA antibody,urinary protein and renal pathology in the AAV-TIPE2 group were significantly lower than those in the AAV-scr group(P<0.01).Conclusion TIPE2 alleviates the disease condition of ALD-DNA induced SLE mice through induction of macrophage polarization to M2 phenotype,which may be used as a promising therapeutic method for ALD-DNA induced SLE mice.

Dvelopment of a DNA-based microarray for detection of human adenovirus serotypes / 军事医学

Objective To develop a chemiluminescence imaging DNA microarray method for simultaneous,quick and accurate detection of serotypes of human adenovirus (HAdV ),namely,HAdV3,HAdV7,HAdV11,HAdV14 and HAdV55.Methods Based on the specific gene sequences in the conserved region of adenovirus from GenBank, oligonucleotide primers and probes were designed and synthesized to prepare the oligonucleotide microarray.The specific genomic sequences were amplified by multiplex PCR method.The multiplex PCR amplification products were hybridized with the specific probes of microarray that was scanned after washing and chemiluminescenceb before the result was analyzed.After optimization of the multiple PCR system,hybridization reactions and conditions of chemiluminescence,the specificity,sensitivity and reproducibility of the chip were evaluated.Results The microarray displayed high sensitivity, specificity and reproducibility.The minimum detection limit of plasmidg DNA was 3 ×103 copies/reaction.The microarray detection results of 38 clinical samples were approximately consistent with those using the direct sequencing method(37 /38).Conclusion A chemiluminescence imaging DNA microarray method for quick,sensitive and specific detection of five serotypes of HAdV is established,which can provide a new means for detecting serotypes of HAdV.

Construction and functional identification of rat Hes1 adenovirus expression vector / 重庆医学

Objective to construct the high titers rat Hes1 adenovirus expression vector (Ad-Hes1).Methods With the rat cDNA as a template,the Hes1 fragment was amplified by PCR,which constructed pShuttle-CMV-Hes1 shuttle plasmid by directly clone.Based on pShuttle-CMV-Hes1,pAdeno-Hes1 virus plasmid was constructed,pAdeno-Hes1 was transfected into 293 cells to package Ad-Hes1,virus titers were determined by modified TCID50.Hes1 was detected by Western blot after Ad-Hes1 infected with H9c2 myocardial cells.Results pShuttle-CMV-Hes1 shuttle plasmid and pAdeno-Hes1 plasmid were constructed successfully,with a general titer of 1.6 × 1011 PFU,Ad-Hes1 can be expressed in H9c2 myocardial cells,and its MOI value was 30.Conclusion Ad-Hes1 is successfully constructedand packaged,thus provide basis for further research on the protection effect of Hes1 on myocardium.

Generation of Insulin-Expressing Cells in Mouse Small Intestine by Pdx1, MafA, and BETA2/NeuroD

BACKGROUND: To develop surrogate insulin-producing cells for diabetes therapy, adult stem cells have been identified in various tissues and studied for their conversion into β-cells. Pancreatic progenitor cells are derived from the endodermal epithelium and formed in a manner similar to gut progenitor cells. Here, we generated insulin-producing cells from the intestinal epithelial cells that induced many of the specific pancreatic transcription factors using adenoviral vectors carrying three genes: PMB (pancreatic and duodenal homeobox 1 [Pdx1], V-maf musculoaponeurotic fibrosarcoma oncogene homolog A [MafA], and BETA2/NeuroD). METHODS: By direct injection into the intestine through the cranial mesenteric artery, adenoviruses (Ad) were successfully delivered to the entire intestine. After virus injection, we could confirm that the small intestine of the mouse was appropriately infected with the Ad-Pdx1 and triple Ad-PMB. RESULTS: Four weeks after the injection, insulin mRNA was expressed in the small intestine, and the insulin gene expression was induced in Ad-Pdx1 and Ad-PMB compared to control Ad-green fluorescent protein. In addition, the conversion of intestinal cells into insulin-expressing cells was detected in parts of the crypts and villi located in the small intestine. CONCLUSION: These data indicated that PMB facilitate the differentiation of mouse intestinal cells into insulin-expressing cells. In conclusion, the small intestine is an accessible and abundant source of surrogate insulin-producing cells.

Epidemiological Correlation between Fecal Adenovirus Subgroups and Pediatric Intussusception in Korea

We investigated the adenoviral etiology and seasonal epidemic trends in intussusception and each adenoviral subgroup. Also we confirmed whether we can use the adenovirus data of Acute Infectious Agents Laboratory Surveillance Report (AIALSR) as an epidemic predictor of intussusception. Patients with intussusception (n = 126), < 5 years old, were enrolled and matched by age and sex with controls suffering acute gastroenteritis without intussusception (n = 106), all recruited at 8 centers. All fecal specimens were assayed for adenovirus, including subgroups A, B, C, E, and F, with reverse transcriptase-polymerase chain reaction (RT-PCR). Adenovirus was detected in 53 cases and 13 controls (P < 0.001). Nonenteric adenoviruses (NEAds) were detected in 51 cases and four controls (P < 0.001). We used Spearman's correlation analysis to analyze the incidence of intussusception and adenoviral epidemic trends, and compared them with fecal and respiratory adenoviral epidemic trends in the AIALSR. The trend of intussusception correlated with total NEAds (r = 0.635; P = 0.011), as did the fecal AIALSR adenovirus trends (r = 0.572; P = 0.026). Among the NEAd subgroups, subgroup C was dominant (P < 0.001), but subgroups B (P = 0.007) and E (P = 0.013) were also significant to intussusception. However, only subgroup C showed a significant epidemic correlation (r = 0.776; P = 0.001) with intussusception. Not respiratory but fecal AIALSR adenovirus trends correlated with the incidence of NEAds and intussusception. We suggest the possibility of using fecal AIALSR adenovirus data as an approximate epidemic predictor of intussusception.

Clinical Impact of Mixed Respiratory Viral Infection in Children with Adenoviral Infection

BACKGROUND: Although adenovirus (ADV) infection occurs steadily all year round in Korea and the identification of respiratory viral coinfections has been increasing following the introduction of multiplex real-time polymerase chain reaction tests, the clinical impact of viral coinfection in children with ADV infection has rarely been reported. MATERIALS AND METHODS: Medical records of children diagnosed with ADV infection were retrospectively reviewed. The enrolled children were divided into two groups based on the identified respiratory viruses: ADV group and coinfection group. Clinical and laboratory parameters were compared between the two groups. RESULTS: In total, 105 children (60 males and 45 females) with a median age of 29 months (range: 0-131 months) diagnosed with an ADV infection were enrolled. Fever (99.0%) was by far the most frequent symptom, followed by respiratory (82.9%), and gastrointestinal (22.9%) symptoms. Upper and lower respiratory tract infections were diagnosed in 56 (53.3%), and 32 (30.5%) children, respectively. Five (4.8%) children received oxygen therapy, and no child died due to ADV infection. Coinfection was diagnosed in 32 (30.5%) children, with rhinovirus (46.9%), and respiratory syncytial virus (21.9%) being the most frequent. The proportions of children younger than 24 months (P <0.001), with underlying medical conditions (P = 0.020), and diagnosed with lower respiratory tract infection (P = 0.011) were significantly higher in the coinfection group than in the ADV group. In a multivariate analysis, only the younger age was significantly associated with coinfection (P <0.001). Although more children in the coinfection group received oxygen therapy (P = 0.029), the duration of fever and hospitalization was not significantly different between the two groups. CONCLUSION: Respiratory viral coinfection with ADV occurred more frequently in children younger than 24 months of age compared with children aged 24 months or older. Respiratory viral coinfection may increase the severity of ADV infection, however, appropriate therapy prevented prolonged hospitalization and poor prognosis due to coinfection.

Pneumonia Caused by Adenovirus Genotype 55 in an Army Recruit Training Center / 대한내과학회지

Adenoviral pneumonia is rare in healthy adults. However, several cases were recently reported in a military training center in South Korea. Adenovirus genotype 3, 6, and 7 are predominant in South Korea. More recently, genotype 55, which emerged sporadically in China, has been reported in South Korea. In this study, we present a case of adenoviral pneumonia caused by genotype 55 in a healthy soldier from an army recruit training center in South Korea.

Study on biocompatibility of Ad-rhBMP-2 transfection on rabbit BMSCs combined with allogeneic DBM / 重庆医学

Objective To observe the biocompatibility of rabbit bone marrow mesenchymal stem cells (BMSCs)combined with allogeneic decalcified bone matrix(DBM)after transfecting adenoviral recombinant human bone morphogenetic protein-2(Ad-rhBMP-2).Methods The rabbit allogeneic DBM material was prepared according to the Ursit method.After transfecting Ad-BMP-2 on rabbit bone marrow mesenchymal stem cells,the immunohistochemical was used to detect the expression of BMP-2 in the transfected cells;after 48 h of transfection,the cells were planted on the allograft DBM,then the scanning electron microscopy was used to observe the cell growth and adhesion condition on material,and the proliferation condition of BMSCs was detected by MTT. Results After 48 h of adenoviral transfection,BMSCs could express BMP-2 successfully.The scanning electron microscopy showed that the cells after transfection adhered well and massively proliferated on DBM material.The MTT assay showed that the prolifer-ation condition of the cells after transfection planted on DBM was normal,which showed no statistically significant difference when compared with the control group (P >0.05).Conclusion The Ad-BMP-2 transfection on BMSCs is well biocompatible to allogene-ic DBM.

Comparing the multiplex RT-PCR method and liquichip technology in the detection of diarrhea-related virus / 中华检验医学杂志

Objective To compare the detection efficiency between multiplex RT-PCR method and liquichip technology for screening the viral etiological agents of diarrhea.Methods The development of the multiplex RT-PCR method.A total of 107 feces samples from patients who suffered from diarrhea and attended to Zhujiang Hospital of Southern University from September 2013 to February 2014 were collected and tested in parallel by both multiplex RT-PCR and xTAG Gastrointestinal Pathogen Panel ( xTAG GPP) for Adenovirus, Norovirus genogroupⅠandⅡ, as well as by both multiplex RT-PCR and monoplex RT-PCR for Astrovirus and Sapovirus.To evaluate the sensitivity and specificity of multiplex RT-PCR, xTAG GPP and monoplex RT-PCR were used as reference.Kappa coefficient test was used to evaluate the consistency among the methods.The detection limit and accuracy of multiplex RT-PCR were evaluated by detection of serial dilution of positive plasmids and products sequencing for the five viral agents.Results The multiplex RT-PCR showed high consistency with xTAG GPP and monoplex RT-PCR, in which Kappa value was 0.885 and 1.000 respectively( P=0.000 ).Compared to xTAG GPP, the sensitivity and specificity of the multiplex RT-PCR were at average of 80.8%( 21/26 ) and 100%( 295/295 ) respectively.The detection limit and accuracy of multiplex RT-PCR were 104 copies /μl-106 copies/μl.Conclusion The high consistency indicated that both the multiplex RT-PCR and xTAG GPP are useful as a special,sensitive, high throughput and rapid diagnostic tools for the detection of the major viral pathogens related to diarrhea in clinical laboratory.