Clinical characteristics of patients with human adenovirus infection undergoing allogeneic hematopoietic stem cell transplantation / 中华内科杂志

Objective:To investigate the clinical characteristics and prognosis of human adenovirus (HAdV) infection in patients with allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:This is a retrospective case series study. Patients who received allo-HSCT and had symptoms of HAdV infection were tested in the Hematology Department at Perking University People′s Hospital from August 2015 to October 2019. Real-time quantitative PCR was used to detect HAdV DNA from 2 728 patients with potential infection. HAdV DNA-positive patients were defined as having HAdV infection. The clinical features of these patients were analyzed, and a case-pair method was used to select patients without HAdV infection as the control group in a 1∶3 ratio. The clinical results of the two groups were compared using Kaplan-Meier and Log-rank testing.Results:A total of 7 119 samples were tested for HAdV, of which 99 samples from 36 patients were positive. Of these patients, 22 developed HAdV viremia, and 24 patients had concurrent infection with another virus. Nineteen patients had fever (53%), 25 had gastrointestinal symptoms (69%), 11 had respiratory symptoms (31%), nine had reduced liver function (25%), and six had nervous system symptoms (17%). Twenty-three patients developed acute graft-versus-host disease of grade 2 or higher. Of all the patients with HAdV infection, nine were treated with cidofovir, seven of whom became HAdV negative and two had invalid treatment. The median follow-up time was 496 (216, 940) d post-HSCT. The overall survival at 5 years post HSCT was 48.4%±9.2% vs. 91.3%±3.5% ( χ2=65.03, P<0.001) in patients with and without HADV, respectively. The non-relapse mortality at 5 years post-HSCT was 40.8%±8.8% vs. 4.0%±2.0% ( χ2=34.17, P<0.001) in patients with and without HADV, respectively. Conclusions:After allo-HSCT, HAdV-infected patients are dominated by gastrointestinal and respiratory symptoms and have an increased risk of combined acute graft-versus-host disease of >2 degrees. Patients with HAdV infection have poor overall survival and high non-relapse mortality.

Clinical features of twenty-three cases of adenoviral encephalitis in children / 中华传染病杂志

Objective:To investigate the clinical features of adenoviral encephalitis (AE), and to provide reference for clinical diagnosis and treatment of adenoviral encephalitis.Methods:From January 2012 to December 2020, 1 185 cerebrospinal fluid (CSF) samples of hospitalized children with suspected central nervous system infection in the Second Affiliated Hospital of Shantou University Medical College were collected for the detection of 22 common respiratory pathogens and common pathogens for encephalitis by polymerase chain reaction. Records of patients with adenovirus positive in CSF were reviewed and relevant clinical manifestations, laboratory tests and imaging examination results were collected for analysis.Results:Among 1 185 CSF samples, 242 samples were positive for viral nucleic acid, with detection rate of 20.4%, including 1.9%(23/1 185) of adenovirus. As for 23 children diagnosed with AE, 18 were male, five were female, with the age of (44.8±35.9) months, ranging from two months and 19 days to 10 years. Of 23 children, 21(91.3%) presented with fever, followed by convulsions (16 cases, 69.6%), headache (four cases, 17.4%), vomiting (11 cases, 47.8%), consciousness change (11 cases, 47.8%) and emotion disturbance (three cases, 13.0%). Among 23 children, eight cases had white blood cell counts (WBC) of (6 to <10)×10 9/L, 10 cases had WBC of (10 to 20)×10 9/L and the white blood cell classification was mainly neutrophils (21 cases, 91.3%), and C reactive protein of 20 cases (87.0%) was in the normal range. Cerebrospinal fluid examination showed that WBC were less than 15×10 6/L in 20 cases (87.0%), and WBC ≥15×10 6/L in three cases, which were up to 500×10 6/L; the protein of 19 cases was in the normal range, the glucose of 15 cases was in the normal range, and the chloride of 19 cases was in the normal range. Among 16 cases with brain magnetic resonance imaging examination, eight cases did not show abnormality, six cases with local meningeal linear enhancement, one case with small intracranial malacia, and one case with extensive intracranial lesions. For 13 cases who received electroencephalogram (EEG) test, seven cases showed normal EEG or marginal state, four cases showed extensive medium and high amplitude slow wave, one case showed spike wave or spike slow wave and one case had both of the above two changes. Among 23 children, 22 cases recovered including one case had secondary epilepsy, and the remaining one case had severe brain dysfunction and was unable to suck when discharged, with an indwelling gastric tube and accompanied by secondary epilepsy. Conclusions:The clinical manifestations and auxiliary examinations of children with AE have no obvious specificity. Most children with AE have a good prognosis, but a small number of them may have serious sequelae.

Classification of adenoviral conjunctivitis based on sequencing and phylogenetic analysis of hexon and fiber / 中华实验眼科杂志

Objective:To classify human adenovirus (HAdV) of adenoviral conjunctivitis by sequencing and phylogenetic analysis of hexon protein and fiber protein.Methods:A total of 256 conjunctival swabs were collected from the inferior conjunctival sac of 256 patients with viral conjunctivitis in Shanghai from January 2015 to August 2017.After DNA extraction, the whole length of hexon and fiber was amplified by PCR to perform gene sequencing and phylogenetic analysis.The study protocol was approved by the Ethics Committee of Shanghai General Hospital (No.2020-202).Written informed consent was obtained from each subject.Results:Among the samples, 89(34.76%) were positive for hexon gene amplification, including 1(1.12%) of HAdV-C1, 7(7.87%) of HAdV-C2, 20(22.47%) of HAdV-B3, 6(6.747%) of HAdV-E4, 23(25.84%) of HAdV-D8, 17(19.10%) of HAdV-D19, and 15(16.85%) of HAdV-D37.In phylogenetic analysis, sequenced hexon gene was clustered with the reference prototype correctly.In fiber phylogenetic analysis, 15 of HAdV-D19 and 1 of HAdV-D37 were cross clustered.Conclusions:The combined sequencing of hexon and fiber can provide abundant and effective biological information for the subtype and pathogenicity analysis of HAdV.

Management of acute respiratory distress syndrome in a child with adenovirus pneumonia: case report and literature review / Manejo da síndrome do desconforto respiratório agudo em criança com pneumonia por adenovírus: relato de caso e revisão da literatura

ABSTRACT Objective: To report the case of a child who developed acute respiratory distress syndrome (ARDS) from a pulmonary infection by adenovirus. Case description: A female patient aged 2 years and 6 months, weighting 10,295 grams developed fever, productive cough and vomiting, later on progressing to ARDS despite initial therapy in accordance with the institutional protocol for ARDS treatment. The child evolved to refractory hypoxemia and hypercapnia, requiring high parameters of mechanical pulmonary ventilation and use of vasoactive agents. In the treatment escalation, the patient received steroids, inhaled nitric oxide (iNO), was submitted to the prone position, started oscillatory high-frequency ventilation (HFOV) and extracorporeal membrane oxygenation (ECMO) was indicated due to severe refractory hypoxemia. During this time, the patient's clinical response was favorable to HFOV, improving oxygenation index and hypercapnia, allowing the reduction of vasoactive medications and mechanical ventilation parameters, and then the indication of ECMO was suspended. The patient was discharged after 26 days of hospital stay without respiratory or neurological sequelae. Comments: Adenovirus infections occur mainly in infants and children under 5 years of age and represent 2 to 5% of respiratory diseases among pediatric patients. Although most children with adenovirus develop a mild upper respiratory tract disease, more severe cases can occur. ARDS is a serious pulmonary inflammatory process with alveolar damage and hypoxemic respiratory failure; Adenovirus pneumonia in children may manifest as severe pulmonary morbidity and respiratory failure that may require prolonged mechanical ventilation. Exclusive pulmonary recruitment and HFOV are advantageous therapeutic options.

RESUMO Objetivo: Descrever paciente que evoluiu com síndrome do desconforto respiratório agudo (SDRA) a partir de infecção pulmonar por adenovírus. Descrição do caso: Paciente de dois anos e seis meses, sexo feminino, peso de 10295 g, que apresentou com quadro de febre, tosse produtiva e vômitos, evoluindo para SDRA. Apesar da terapêutica inicial em conformidade com o protocolo institucional de tratamento da SDRA, a criança evoluiu para hipoxemia e hipercapnia refratárias, necessitando de elevados parâmetros de ventilação pulmonar mecânica e utilização de agentes vasoativos. No escalonamento da terapêutica, a paciente recebeu terapias adjuvantes, foi iniciada ventilação oscilatória de alta frequência (VOAF) e indicada oxigenação por membrana extracorpórea (OMEC) pela hipoxemia grave refratária. Nesse ínterim, a paciente apresentou resposta clínica favorável à VOAF, melhorando do quadro ventilatório e possibilitando a redução das medicações vasoativas e dos parâmetros de ventilação mecânica. A paciente recebeu alta hospitalar após 26 dias de internação, sem sequelas respiratórias ou neurológicas. Comentários: As infecções por adenovírus ocorrem principalmente em lactentes e crianças com menos de cinco anos de idade e representam de 2 a 5% das doenças respiratórias entre os pacientes pediátricos. Embora a maioria das crianças com infecção por adenovírus desenvolva doença leve do trato respiratório superior, casos mais graves podem ocorrer com comprometimento do trato respiratório inferior. A pneumonia por adenovírus em crianças pode se manifestar com morbidade pulmonar grave e insuficiência respiratória com risco de vida, o que resulta na necessidade de suporte mecânico prolongado. O recrutamento pulmonar exclusivo pela VOAF pode ser uma opção terapêutica útil.

Human adenovirus type 7 associated respiratory infections and its laboratory examination / 中华检验医学杂志

In recent years , respiratory tract infectious diseases and public health events caused by human adenovirus type 7(HADV-7) had been rising.Especially in Asia, the outbreaks of acute respiratory illness caused by HADV-7, accounting for more than 60%in the world, had brought great influence to the public health.In order to prevent the occurrence of HADV-7 epidemic and improve the diagnostic performance of laboratory , the research progress of HADV biological features and typing , HADV-7 epidemiological characteristics , clinical and differential diagnosis and laboratory examination methodologies were reviewed in this paper.

Adenovírus e rotavírus em águas superficiais do córrego Ribeirão Preto, São Paulo, Brasil / Adenovirus and rotavirus in surface water of the Ribeirão Preto stream, São Paulo, Brazil

Com o desenvolvimento humano, os recursos hídricos sofrem modificações derivadas da expansão demográfica e principalmente da falta de saneamento básico, que interferem diretamente na qualidade desses recursos com a introdução de substâncias químicas tóxicas e bioagentes patogênicos. Os vírus entéricos de veiculação hídrica, como adenovírus e rotavírus, merecem destaque considerando seus impactos na saúde pública. O objetivo deste estudo foi avaliar a presença de adenovírus e rotavírus em águas superficiais do Córrego Ribeirão Preto, SP. A coleta foi realizada em 11 pontos distintos, da nascente até a confluência com o Rio Pardo. O Teste de ELISA foi o método utilizado para a detecção dos vírus (RIDASCREEN® Adenovirus, RIDASCREEN® Rotavirus - R-Biopharm). De acordo com os resultados obtidos, 73% das amostras foram positivas para adenovírus, enquanto que 36% foram positivas para rotavírus. Considerando a virulência desses vírus entéricos e a vulnerabilidade de indivíduos imunossuprimidos, idosos e crianças, é preocupante a contaminação por esses agentes patogênicos de veiculação hídrica amplamente disseminados em águas superficiais.

Water resources have been suffering with the changes in demographic expansion and the lack of basic sanitation. These factors directly interfere with the quality of these resources due to the discharge of toxic chemicals and pathogenic bio-agents. The waterborne enteric viruses, such as rotavirus and adenovirus, deserve attention considering its impact on public health. The aim of this study was to evaluate the presence of adenovirus and rotavirus in surface water of Ribeirão Preto stream. Samplings were conducted in 11 different points from the spring to the confluence with the Rio Pardo. The ELISA test was carried out for the detection of viruses (RIDASCREEN® adenovirus, rotavirus RIDASCREEN® - R-Biopharm). According to the results, 73% of the samples were positive for adenovirus, while 36% were positive for rotavirus. Given the virulence of these enteric viruses and the vulnerability of immunosuppressed individuals, i.e. the elderly and children, contamination with these waterborne pathogens widely disseminated in surface water is concerning

Advance research of p53 gene therapy combination with transcatheter arterial chemoembolization for hepatocellular carcinoma / 国际外科学杂志

About the malignant tumor in China,the hepatocellular carcinoma mortality is second only to lung cancer and serious threat to the life and health of the masses.Furthermore,because most patients has been in advanced cancer during medical treatment,so had lost the chance of one-stage surgical resection.However,the sensitivity of hepatocellular carcinoma to chemotherapy,radiotherapy and other treatments are poor.Transcatheter arterial chemoembolization is the main method of the treatment for patients those have lost the chance of operation,though the clinical effect is significant,the inadequate is also presence,such as tumor necrosis,incompletely clear,residual tumor nidus and the damage of the immune function after operation.Recombinant adenovirus p53 gene can validly infect tumor cells,transcription and expression of p53 protein,it also regulate the expression of related genes inhibiting tumor cell growth and induce cell apoptosis directly or indirectly.It is increasingly highly attention that recombincanting rAd-p53 with transcatheter arterial chemoembolization(TACE) to treat hepatocellular carcinoma.In order to evaluate the clinical value and promising future,we will make a brief summary for the research progress in this area in recent years.

Clinical study of endoscopic intratumor injection of recombinant human adenovirus p53 (rAd-p53) combined with esophageal stent implantation in the treatment of advanced esophageal cancer / 中国基层医药

Objective To compare the efficacy and incidence of complications of stenting alone and intratumoral injection of recombinant human adenovirus p53 combined with stent implantation in the treatment of advanced esophageal cancer patients.Methods 2 4 cases with advanced esophageal cancer,who could only eat semi liquid,liquid food or could not eat,were randomly divided into control group(A group) and treatment group (B group).12 cases in A group were treated with esophageal stent implantation,12 cases in B group were given endoscopic intratumor injection of recombinant human adenovirus p53 combined with food tract stent implantation.Results The rate of improvement of dysphasia had no significant difference in survival period:the treatment group survival time of 3.2-38 months,mean (27.0 ± 2.3) months,while the control group survival time of 2.8-27months,an average of (14.7 ± 8.0) months,the difference between the two groups was significant (t =3.521,P ＜ 0.05).The incidence rate of restenosis in treatrnent group was 8.3％,that in control group was 41.7％.There was significant difference (x2 =7.532,P ＜0.05).Conclusion Endoscopic intratumor injection of recombinant human adenovirus p53 combined with esophageal stent implantation in the treatment of advanced esophageal carcinoma can prolong the survival time of patients with esophageal stent implantation,and to reduce the incidence of restenosis.

Clinical and epidemiological aspects related to the detection of adenovirus or respiratory syncytial virus in infants hospitalized for acute lower respiratory tract infection / Aspectos clínicos e epidemiológicos relacionados à detecção de adenovírus ou vírus sincicial respiratório em crianças hospitalizadas por doença aguda do trato respiratório inferior

OBJECTIVE: To characterize and compare clinical, epidemiological, and laboratory aspects ofinfants with acute lower respiratory infection (ALRI) associated with the detection of adenovirus(ADV) or respiratory syncytial virus (RSV). METHODS: A preliminary respiratory infection surveillance study collected samples of nasopharyngeal aspirate (NPA) for viral research, linked to the completion of a standard protocol, from children younger than two years admitted to a university hospital with ALRI, between March of 2008 and August of 2011. Polymerase chain reaction (PCR) was used for eight viruses: ADV, RSV, metapneumovirus, Parainfluenza 1, 2, and 3, and Influenza A and B. Cases with NPA collectedduring the first 24 hours of admission, negative results of blood culture, and exclusive detection of ADV (Gadv group) or RSV (Grsv group) were selected for comparisons. RESULTS: The preliminary study included collection of 1,121 samples of NPA, 813 collected in thefirst 24 hours of admission, of which 50.3% were positive for at least one virus; RSV was identifiedin 27.3% of cases surveyed, and ADV was identified in 15.8%. Among the aspects analyzed inthe Gadv (n = 58) and Grsv (n = 134) groups, the following are noteworthy: the higher meanage, more frequent prescription of antibiotics, and the highest median of total white blood cellcount and C-reactive protein values in Gadv. CONCLUSIONS: PCR can detect persistent/latent forms of ADV, an aspect to be considered wheninterpreting results. Additional studies with quantitative diagnostic techniques could elucidatethe importance of the high frequency observed. .

OBJETIVO: Caracterizar e comparar aspectos clínicos, epidemiológicos e laboratoriais delactentes com evidências de infecção aguda do trato respiratório inferior (IATRI) associada à detecção do adenovírus (ADV) ou do vírus sincicial respiratório (VSR). MÉTODOS: Um estudo preliminar de vigilância de infecções respiratórias desenvolveu coleta de aspirado nasofaríngeo (ANF) para pesquisa viral, vinculada ao preenchimento de protocolo padrão, de menores de dois anos internados com quadro de IATRI em hospital universitário, entre março de 2008 e agosto de 2011. Utilizou-se técnica da reação em cadeia da polimerase (PCR) para oito vírus: ADV, VSR, metapneumovírus, parainfluenza 1, 2 e 3 e influenza A e B. Foram selecionados para comparações os casos com ANF coletado nas primeiras 24 horas da admissão, resultado de hemocultura negativo e detecção exclusiva de ADV (grupo Gadv) ou VSR (grupo Gvsr). RESULTADOS: O estudo preliminar incluiu coleta de 1.121 amostras de ANF, sendo 813 coletadas nas primeiras 24 h da admissão, das quais 50,3% foram positivas para ao menos um dos vírus, com VSR em primeiro lugar, em 27,3%, e ADV em segundo, em 15,8% dos casos pesquisados. Dentre os aspectos analisados nos grupos Gadv (n = 58) e Gvsr (n = 134), destacaram-se a média da idade mais elevada, maior frequência da prescrição de antibióticos e medianas mais elevadas para contagem total de leucócitos e valores da proteína C-reativa no Gadv. CONCLUSÕES: A PCR utilizada pode detectar formas persistentes/latentes de ADV, aspecto aser considerado ao interpretar os resultados. Estudos complementares com técnicas diagnósticas quantitativas, por exemplo, poderiam evidenciar a importância da elevada frequência verificada. .

Construction of tissue engineering adult cardiac myocytes / 中国组织工程研究

BACKGROUND:Construction of tissue engineering adult cardiac myocytes has been a new research hotspot in cardiovascular fields. OBJECTIVE:To explore a simple, fast method for the separation of adult rat cardiomyocytes, and preliminarily explore the construction methods of tissue engineering adult cardiac myocytes. METHODS:Segmented enzymatic digestion method was used to isolate cardiac myocytes from adult rats. Subsequently, cardiac myocytes were transfected with adenovirus and liposome-mediated red fluorescent protein gene. Construction efficiency of tissue engineering cells was qualified using inverted fluorescence microscopy and flow cytometry. Final y, cardiac myocytes were transfected with adenovirus-mediated hypoxia-inducible factor-1a, and the expression of hypoxia-inducible factor-1a was examined by western blot analysis. RESULTS AND CONCLUSION:A lot of cardiac myocytes were col ected using the segmented enzyme digestion method. Flow cytometric analysis showed that the survival of adult cardiomyocytes was (87.03±0.70)%. Compared with liposomal transfection (transfection efficiency was 0), adenovirus infection efficiency was (70.31± 1.39)%, and the cells expressed red fluorescence under fluorescence microscope. After 4 days of adenovirus infection, transfected cells expressed hypoxia-inducible factor-1a protein. These results showed that segmented enzyme digestion is a fast way to isolate adult cardiac myocytes, and recombinant adenovirus vector is a good vector to transfect cardiac myocytes.

The aetiological analysis of common viruses of acute lower respiratory infection among the hospitalized children 0 to 7 years old in Nantong of Jiangsu / 中国基层医药

Objective To investigate the infection status and clinical features of common viruses in acute lower respiratory infection (ALRI) among the hospitalized children 0 to 7 years old in Nantong of Jiangsu.Methods 1 376 swab samples from pharynx nasalis in the ALRI inpatients 0 to 7 years old were collected.The human respiratory syncytial virus (RSV),adenovirus (ADV),influenza virusA,B (IVA,B),parainfluenza virus Ⅰ ～ Ⅲ (PIV Ⅰ ～ Ⅲ)were detected by direct immunofluorescence assay,and the results were analyzed.Results In 1 376 respiratory tract samples,there were 577 cases(41.93％) of positive samples.In all positive samples,there were 376 cases of RSVpositive (65.16％),42 ADV-positive (7.28％),63 IVA-positive (10.92％),24 IVB-positive (4.16％),20 PIV Ⅰ-positive(3.47％),19 PIV Ⅱ-positive (3.29％),108 PIV Ⅲ-positive (18.72％),68 cases with mixed infection (11.79％) [two virus-positive ones in 59 cases (86.76％),three virus-positive ones in 9 cases (13.24％)].In different age group,the highest positive rate was in 0 ～ 6-month-old group(53.32％),with the lowest in 5-7 years old group(6.90％).Virus detection rate was higher in March 2012 (58.67％),December 2012 (53.33％),and January 2013(53.63％)than the rest months,including the lowest June 2012(33.33％).Bronchiolitis virus detection rate was the highest(69.23％)among ALRI.Conclusion The virus is major pathogen of children 0 to 7 years old with ALRI in Nantong of Jiangsu,and with difference among different ages,seasons and diseases.Infants and young children are the main affected population.

Research on induced apoptosis of recombinant human adenovirus-p53 injection in patients with oral leukoplakia / 肿瘤研究与临床

Objective To probe the biological effect of multiple intraepithelial injections of recombinant adenovirus-p53(rAd-p53)on inducing the apoptosis in patients with dysplastic oral leukoplakia.Methods 18 patients clinically and histopathologically diagnosed as dysplastic oral leukoplakia were recruited for this study.Intraepithelial injections of rAd-p53 were administered.Immunohistochemistry was used to examine the protein expression of p53 and bcl-2.TUNEL was performed to detect apoptotic cells.Results In the post-treatment patients,p53 protein expression were significantly enhanced(100 ％,18/18),yet bcl-2 protein presented low expression(17 ％,3/18).Statistical analysis revealed the expression of p53 protein had a negative correlation with that of bcl-2 protein(r =-0.837,P ＜ 0.01).15 post-treatment samples (83 ％)were detected obvious apoptotic cells,especially in the samples that were strong p53-positive(r =0.684,P ＜ 0.01).Conclusion Intraepithelial injections of rAd-p53 can induce apoptosis for patients with dysplastic oral leukoplakia.It may be a promising treatment for oral leukoplakia.

Infection with adenovirus-mediated luciferase reporter gene in mesenchymal stem cells and bioluminescence imaging / 中华核医学与分子影像杂志

Objective To construct adenovirus vector containing firefly luciferase reporter gene (AdLuc) and infect bone marrow mesenchymal stem cells (BMSC),then to take bioluminescence imaging in vitro and in vivo for identification.Methods The luciferase gene was amplified with PCR from psiCHECK-2 plasmid and cloned into the adenoviral shuttle vector (pShuttle-CMV).It was confirmed by Nhe Ⅰ/Xba Ⅰ digestion and sequencing.PShuttle-CMV-Luc and backbone vector (pAdeno) were homologous recombined.Then the recombinant plasmid was packaged in HEK293 cells and the virus titer was detected.The BMSC were infected by the recombinant adenovirus.The bioluminescence imaging in vitro was performed to determine the best multiplicity of infection (MOI),and the relationship between bioluminescence intensity and MOI was analyzed by curve fitting regression analysis.Viability was evaluated via Trypan blue staining.The transfected BMSC (l× 106) were implanted into the muscles of forelimb of SD rats,and then tracked by bioluminescence imaging in vivo.Cell viability was compared using two-way repeated measures analysis of variance between groups.Results Enzyme digestion and sequence analysis indicated that Ad-Luc was successfully constructed.The virus titer was 1 × 1010 plaque forming unit (PFU)/ml.The bioluminescence detection in vitro showed that Ad-Luc could infect BMSC high efficiently to express luciferase and the best MOI was 50.The bioluminescence intensity enhanced with increase of MOI (R2 =0.98).No statistically significant difference was found in cell viability between transfected and untransfected BMSC at 1,3,5,7 d.The cell survival rates were (92.5±2.3)％ vs (94.1±1.8)％,(91.4±0.9)％ vs (92.7±2.0)％,(92.1±1.6)％ vs (93.3± 2.4) ％,(91.9 ± 1.5) ％ vs (93.0 ± 3.1) ％,respectively (F =4.38,P ＞ 0.05).The bioluminescence imaging in vivo showed that BMSC survived 1,3,7 d after implantation.However,bioluminescence signal decreased gradually over time.Conclusion It is feasible to apply the optical reporter gene imaging for tracing transplanted stem cells in vitro and in vivo due to the effective transformation of luciferase reporter gene into BMSC by adenovirus vector.

Uso de ganciclovir 0,15 por cento gel para tratamento de ceratoconjuntivite adenoviral / Ganciclovir 0.15 percent ophthalmic gel in the treatment of adenovirus keratoconjuntivitis

OBJETIVOS: Determinar a efetividade e a toxicidade do ganciclovir 0,15 por cento gel no tratamento de ceratoconjuntivites adenovirais e na prevenção de complicações tais como infiltrados corneanos, membranas ou pseudomembranas conjuntivais. MÉTODOS: Ensaio clínico duplo-cego, intervencionista, randomizado. Os 33 pacientes com diagnóstico clínico de ceratoconjuntivite adenoviral com início dos sintomas há menos de cinco dias foram randomizados em dois grupos: Grupo 1 (tratamento) com 19 pacientes que usaram ganciclovir e Grupo 2 (controle) com 14 pacientes que usaram lágrima artificial sem conservante. Todos pacientes responderam a um questionário de sinais e sintomas e foram submetidos a um exame oftalmológico. No 6º dia de tratamento responderam ao mesmo questionário por telefone e no 10º dia foram reavaliados pelo mesmo examinador e responderam novamente ao questionário. Os sinais e sintomas foram comparados. Para análise estatística foi utilizado os testes T de Student, Mann-Whitney e Wilcoxon, com significância estatística p<0,05. RESULTADOS: Tendência de melhor resposta no grupo tratamento em relação à percepção pelos pacientes, além da melhora mais rápida desse grupo em relação ao grupo controle (p=0,26). Houve menor transmissão para o olho adelfo (p=0,86) e para pessoas do convívio (p=0,16) no grupo tratamento. Comparando os dois grupos não houve diferença estatística em relação aos sintomas e sinais da conjuntivite. Comparando isoladamente cada grupo entre o pré-tratamento e no decorrer do tratamento, observou-se melhora estatisticamente significativa da dor, prurido e fotofobia apenas no grupo tratamento. Ganciclovir não mostrou toxicidade e teve maior tolerância pelos pacientes. Não houve diferença significativa no aparecimento de complicações da conjuntivite entre os dois grupos. CONCLUSÕES: O estudo evidenciou uma tendência à melhora mais rápida dos sinais e sintomas dos pacientes tratados com ganciclovir em relação ao grupo ...

PURPOSE: To evaluate the efficacy and the toxicity of 0.15 percent ganciclovir gel in the treatment of adenoviral conjunctivitis and in preventing ocular complications after adenoviral conjunctivitis, such as corneal infiltrates and pseudomembranes. METHODS: Double blind, interventional and randomized clinical trial. Thirty-three patients with clinical diagnosis of adenoviral conjunctivitis with onset of symptoms for five or less days were randomized in two groups: Group 1 (treatment) with 19 patients used ganciclovir gel and Group 2 (control) with 14 patients used artificial tears without preservative. Patients answered a questionnaire of signs and symptoms and were submitted to an ophthalmologic exam. On the 6th and 10th days of treatment they answered the same questions and were re-examined by the same ophthalmologist. Signs and symptoms were compared. T Student, Mann-Whitney e Wilcoxon tests were used to statistical analysis. RESULTS: Trend of better response in the treatment group in relation of patients' perception, besides faster improvement of this group compared to the control group (p=0.26). There were lower transmission to the fellow eye (p=0.86) and to people living together (p=0.16) in the treatment group. No statistical difference related to signs and symptoms of conjunctivitis were found comparing both groups. We observed statistical difference in pain, itch and photophobia only in the treatment group, comparing each group alone. No toxicity and more tolerance of the ganciclovir were observed. There was no statistical difference in the ocular complications after conjunctivitis between both groups. CONCLUSIONS: This study showed trend of better and faster response of the signs and symptoms of the patients treated with ganciclovir compared with the control group, but with no statistical significant. These results need to be confirmed by additional studies, with more patients and longer follow-up. Clinical Trails.gov: NCT01349452.

Oncolytic adenovirus expressing lipocalin 2 suppresses the growth of transplantation tumors of colonic carcinoma / 中华普通外科杂志

Objective To investigate the antitumor activity of the oncolytic adenovirus expressing lipocalin 2 gene for colorectal cancer in vivo.Methods BALB/C nude mice subcutaneously inoculated by SW620 cells and grown tumors were treated with injection of ZD-55 virus, Ad-lipocalin 2 virus and ZD55- lipocalin 2 virus respectively.The weight of implanted tumors and the tumor inhibition rate were calculated to evaluate the anti-tumor effect.Cell apoptosis was determined by TUNEL and the protein expression of VEGF and MVD were determined with immunohistochemistry.Results ZD55-lipocalin 2 inhibited the growth of transplanted tumor more significantly than ZD-55 virus and Ad-lipocalin 2 virus ( P ＜ 0.05 ).Tumor cell apoptosis was upregulated and the MVD reduced significantly in ZD55-lipocalin 2 group in contrast to the other two groups (P ＜0.05).Conclusions ZD55-lipocalin 2 induces apoptosis of colorectal tumor cells and inhibits tumor microvascular formation, slowing down the growth of transplantation tumors.

Inhibitory effect of Ad-hTERTp-HSV-TK/GCV system on mice ascites production planted by liver cancer cell line / 肿瘤研究与临床

Objective To observe the effect of Ad-bTERTp-HSV-TK/GCV system on malignant ascites of mice and probe into its mechanism of action.Methods The SX1 inbred strain mice were injected with H22 cell line of liver cancer and were divided into 4 groups at random.The mice in each group were given corresponding treatment after 48 hours.The production of ascites and survival period were evaluated. The apoptosis rates of tumor cells were detected by FCM.Morphological changes of tumor cells were studied by electromicroscope.Results Compared with other groups.Ad-hTERTp-HSV-TK/GCV Can obviously inhibit the production of ascites(P<0.01),prolong the survival period (P<0.01),and apoptosis rate in this group (27.12±2.12)% was significantly higher than that in other groups.No obvious side effect Was found during the treatment.Conclusion Ad-hTERTp-HSV-TK/GCV system Can inhibit production of ascites and prolong the survical period of mice by inducing apoptosis of hepatoma cells,which is a safe and feasible treatment for hepatoma therapy.

Effects of wt-p53, BT-1 and CM-CSF genes cotransfection on proliferation and immunogenicity of SH-SY5Y cell lines / 国际儿科学杂志

Objective To observe the effeet of cotransduction of wt-p53,B7-1 and GM-CSF genes mediated by recombinant adenovirus on cell growth, apoptosis and immunogenicity of neuroblastoma cell line SH-SY5Y, Methods We transfected SH-SYSY cells separately with recombinant adenovirus, recombinant adenovirus mediating human wild type p53(wt-p53) gene and BB-102.We separately named them with SH/Ad,SH/p53 and SH/BB-102 group.The protein expression of p53, B7-1 and GM-CSF were measured by Western blot, FCM and enzyme linked immunosorbent assay. Cell number was counted and growth curves were drawed. Cell apoptosis was tested by FCM. Lymphocyte proliferation and cytokine secretory was evaluated by mixed lymphocyte culturing. Results After transection with BB- 102, p53, B7-1, and GM-CSF, high-efficiency expression of target genes were found. Growth of the cells was inhibited, apoptosis was induced in the SH/p53 and SH/BB-102 groups. Proliferation of lymphocytes was stimulated and cytokine increased visorously in the SH/BB-102 group. Condusions After transfection with BB-102, growth of the SH-SY5Y cells was inhibited,apoptosis was induced,and immunogenicity was greatly enhanced.

Polarization of protective immunity induced by replication-incompetent adenovirus expressing glycoproteins of pseudorabies virus

Replication-incompetent adenoviruses expressing three major glycoproteins (gB, gC, and gD) of pseudorabies virus (PrV) were constructed and used to examine the ability of these glycoproteins to induce protective immunity against a lethal challenge. Among three constructs, recombinant adenovirus expressing gB (rAd-gB) was found to induce the most potent immunity biased to Th1-type, as determined by the IgG isotype ratio and the profile of the Th1/Th2 cytokine production. Conversely, the gC-expressing adenovirus (rAd-gC) revealed Th2-type immunity and the gD-expressing adenovirus (rAd-gD) induced lower levels of IFN-gamma and IL-4 production than other constructs, except IL-2 production. Mucosal delivery of rAd-gB induced mucosal IgA and serum IgG responses and biased toward Th2-type immune responses. However, these effects were not observed in response to systemic delivery of rAd-gB. In addition, rAd-gB appeared to induce effective protective immunity against a virulent viral infection, regardless of whether it was administered via the muscular or systemic route. These results suggest that administration of replication-incompetent adenoviruses can induce different types of immunity depending on the expressed antigen and that recombinant adenoviruses expressing gB induced the most potent Th1-biased humoral and cellular immunity and provided effective protection against PrV infection.

Construction of cancer-specific replication-competent adenovirus vector expressing IL-18 gene and its cytotoxic effect on hepatocellular carcinoma cells in vitro / 肿瘤

Objective: To construct conditional replication adenovirus vector CNHK600-IL-18, in which the IL-18 expression was driven by hypoxia promoter and adenovirus replication was regulated by human telomerase reverse transcriptase (hTERT) promoter. Specific replication ability and the killing effect of CNHK600-IL-18 on hepatocellular carcinoma (HCC) cells were investigated. Methods: Viral replication experiments (TCID50 method) were performed to evaluate the selective replication ability of pSG600 and CNHK600-IL-18 during 96 h in three different hepatocellular carcinoma cell lines. MTT assay was applied to evaluate the killing efficacy of pSG600 and CNHK600-IL-18 on the three HCC cell lines at different MOI. BJ cells were infected with CNHK600-IL-18. The supernatant was collected and the level of IFN-γ secreted by T cells was measured. Results: Western blot showed that IL-18 was detected in SMMC-7721 cells after CNHK600-IL-18 infection. The supernatant of CNHK600-IL-18 -infected BJ cells induced T cells to secret IFN-γ. CNHK600-IL-18 replications in HCC cell lines BEL-7404 and SMMC-7721 were 31 658 and 125 396 after 96-h incubation, respectively, which were 400 and 1587 folds as much as in BJ cell line. Conclusion: CNHK600-IL-18 selectively replicates in HCC cells and generated bioactive IL-18 with high efficacy. CNHK600-IL-18 has an obvious tumor-killing effect against HCC.

Ectopic bone formation using bone morphogenetic protein 2 adenoviral vector / 吉林大学学报(医学版)

Objective To evaluate the osteoinductive activity of recombinant adenoviral vector carrying the human BMP-2 gene(AdBMP-2) and investigate the mechanism of bone formation induced by the bone morphogenetic protein 2(BMP-2).Methods AdBMP-2 infected rabbit bone messenchymal stem cells(BMSC) were cultivated in vitro.The expression of hBMP-2 was determined by in situ hybridization and immunohistochemical analysis,and ALP was determined by enzyme cytochemistry 7 d after infection.At 21th day,Alizarin red S staining was used to evaluate calcium-rich deposits by cells transfected in culture.30 ?L AdBMP-2 solution(2?10~(12)pfu?mL~(-1)) was injected into the muscle of the left hind limb of the nude rat,and 30 ?L Ad? gal solution(2?10~(12)pfu?mL~(-1)) was injected into the muscle of the right hind limb as control.Animals were sacrificed at 20th day following injection to investigate the bone formation histologically and the expression of BMP-2 in the specimens of the injection sites by immunohistochemistry and in situ hybridization method.Results The expressions of BMP-2 and ALP in the transduced BMSC were confirmed at 7 th day after infection.A deeply stained mineralized layer was evident at 21th day in transduced cells.In vivo experiment,BMP-2 gene therapy induced mesenchymal monocytes chemotaxis,proliferation, differentiation into chondrocytes,with subsequent endochondral bone formation.At the same time,hBMP-2 expression was observed in the chondrocytes,osteoblasts and mesenchymal monocytes.(Conclusion AdBMP-2) posseses the osteoinductive activity in vitro and in vivo.