Objective To study characters of pathology associated with functional alteration of primary brainstem injury (PBI) at different injury severities in rabbits .Methods Animal model of graded PBI was produced using rabbits .Animals were di‐vided into five groups ,group Ⅰ to Ⅳ with an increase of impact power ,10 cases in each group ,and the control group with 5 cases . The pathology characters of PBI were investigated combining dissection observation with unaided eye ,tissue HE histochemical stai‐ning and electron microscope .Results Slight brainstem injury were observed in group Ⅰ ,and pathological results showed regional subarachnoid hemorrhage (SAH) ,stripping of regional cerebral pia mater ,a few petechial hemorrhage in surface ,nerve cell edema , normal medulla sheath and axon roughly .The brainstem injuries were apparent in group Ⅱ ,and the pathological changes indicated SAH in sheet ,petechial and sheeted hemorrhage in surface ,and slight swelling and vacuoles in nerve cells .The brainstem injuries were observed obviously in group Ⅲ ,exhibiting thick SAH ,petechial and sheeted hemorrhage in surface and inside ,degeneration of nerve cells ,abruption of axon ,and atrophy of axoplasm .Eight of ten animals died of respiratory depression induced by brainstem in‐jury in group Ⅳ ,presenting thick hemorrhage in subarachnoid surrounding brainstem ,the whole brainstem injured ,microscopically with multiple small hemorrhage ,nerve cells only residual nuclei ,myelin lamellar severe stratification and fracture ,and axonal tran‐section ,disintegration .No abnormal pathological changes were shown in control group .Conclusion The impacts to brainstem with higher powers lead to more manifest functional and more severe pathological changes ,with an alternation of injury location from surface to deep .

Objective To establish a rapid assay with high sensitivity and specificity based on the sequences for group specific gene (GS) and pathogenicity island pag A gene.Methods The PCR primers and probes were designed after the whole sequence was systemically analyzed with bio-informafion tools and blasted with Genebank database.The amplicons were inserted into plasmids so that they could be used as the standard templates to evaluate the sensitivity of the diagnostic system.This assay was based on TaqMan probes and portable Smartcycle PCR machine.Results The detection level was approximately 100 copies per reaction.There was no cross-reaction with other species of Bacillus.This assay could be completed in one hour in laboratory.Conclusion The duplex TaqMan PCR assay could be used to detect Bacillus anthracis rapidly with high sensitivity and specificity.

BACKGROUNDGastroesophageal reflux disease with extra-esophageal symptoms, especially those with respiratory distress was attracting more and more attention. The related mechanisms were still in controversy. The purpose of the work was to explore airway inflammation triggered by gastroesophageal reflux.

METHODSSixteen Sprague-Dawley rats were used as study group and 9 as control. In the study group, a plastic extender with a trumpet-shaped distal end was inserted into the lower esophagus to dilate the cardia, the pylorus was ligated. One ml of 0.1 mol/L hydrochloric acid was injected into the stomach. While a simple laparotomy was performed for control animals. All animals from two groups were sacrificed 24 hours after operation. Then tracheotomy was carried and the bronchoalveolar lavage fluid was collected in all animals. Cells in the fluid were counted and levels of interleukin (IL)-5, -6, -8 in it were measured.

RESULTSCompared with control group, the study group presented a neutrophil pattern of airway inflammation and an elevated concentration of IL-5, -6, -8 with no significant difference regarding eosinophil count.

CONCLUSIONThe gastroesophageal reflux-triggered airway inflammation is characterized by a neutrophilic airway inflammation which differed from that caused by asthma, and enhanced levels of IL-5, -6 and -8, which are similar to that caused by asthma.

Animals ; Asthma ; etiology ; Bronchoalveolar Lavage Fluid ; immunology ; Disease Models, Animal ; Female ; Gastroesophageal Reflux ; complications ; Inflammation ; etiology ; Interleukin-5 ; analysis ; Interleukin-6 ; analysis ; Interleukin-8 ; analysis ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the features and duration of viral nucleic acid shedding in children with influenza A.

METHODSThe clinical data of 90 children with influenza A with positive influenza A virus nucleic acid in nasopharyngeal swab detected by PCR were collected, and these children were divided into simple influenza A group (n=10), influenza A-pneumonia group (n=61), influenza A-nervous system damage group (n=10), and influenza A-underlying disease group (n=9). A retrospective analysis was performed for clinical features, treatment process, duration of viral nucleic acid shedding, and prognosis.

RESULTSThe most common symptoms in these children were fever (89/90, 99%), cough (89/90, 99%), running nose (69/90, 77%), shortness of breath (26/90, 29%), and myalgia (23/90, 26%). The mean duration of viral nucleic acid shedding in 90 children was 9.4±2.9 days. The simple influenza A group had a significantly shorter duration of viral nucleic acid shedding than the influenza A-pneumonia, influenza A-nervous system damage, and influenza A-underlying disease groups (p<0.05), while there were no significant differences between the influenza A-pneumonia, influenza A-nervous system damage, and influenza A-underlying disease groups (p>0.05). The children who received antiviral therapy within 48 hours after disease onset had significantly shorter duration of viral nucleic acid shedding and time to body temperature recovery than those who received antiviral therapy more than 48 hours after disease onset (p<0.05). Of all the children with body temperature recovery, 83% still tested positive for viral nucleic acid.

CONCLUSIONSComplications, underlying diseases, and timing of antiviral therapy are influencing factors for the duration of influenza A virus nucleic acid shedding, and whether body temperature returns to normal cannot be used to decide whether to continue antiviral therapy.

Child ; Child, Preschool ; Female ; Fever ; etiology ; Humans ; Infant ; Influenza A virus ; isolation & purification ; Influenza, Human ; virology ; Male ; Nucleic Acids ; metabolism ; Retrospective Studies ; Time Factors ; Virus Shedding

AIMTo study the effect of volume expansion by 0.9% and 1.8% sodium solution on cardiac-renal reflex activity and the role of cardiac-renal reflex in the regulation of integrated function.

METHODS18 health pentobarbital-anaesthetized rabbits were divided evenly into 2 groups at random, bilateral sino-aortic denervation, intubated via right jugular vein to monitor CVP, left renal nerve separated and ending sectioned to record ERSNA, bilateral ureter intubated to collect urine, right femoral intubated to get blood sample. 15% of whole body blood volume of 0.9% and 1.8% sodium solution were injected via jugular vein 10 ml per minute respectively. The CVP, ERSNA, bilateral urine volume and coefficient of sodium excretion were measured before treated, during treated, one minute, five minutes and ten minutes after treated.

RESULTSVolume expansion by 0.9% and 1.8% sodium solution respectively resulted in the increase of CVP by 64.00% +/- 15.56% and 77.00% +/- 23.74%; the decrease of the frequency of ERSNA by 44.00% +/- 13.64% and 63.00% +/- 12.49%, the average burst time of ERSNA by 37.00% +/- 16.49% and 31.00% +/- 10.69%, the increase of average interval of ERSNA bursts by 60.00% +/- 18.38% and 68.00% +/- 27.04%; the increase of urine volume by 158.00% +/- 28.10% and 640.00% +/- 155.39% in left kidney, 192.00% +/- 32.26% and 1343.00% +/- 429.95% in the right; the increase of coefficient of sodium excretion by 132.00% +/- 35.23% and 376.00% +/- 121.72% in the left, 300.00% +/- 76.99% and 856.00% +/- 261.48% in the right.

CONCLUSIONVolume expansion by different solution stimulate the receptors of cardiopulmonary and regulate the water and sodium excretion of the kidney by the cardiac-renal reflex to modulate the stabilization of blood volume.

Animals ; Blood Volume ; drug effects ; physiology ; Central Venous Pressure ; Heart ; drug effects ; innervation ; Kidney ; drug effects ; innervation ; Rabbits ; Reflex ; Saline Solution, Hypertonic ; pharmacology

Humans ; Liver Diseases ; complications ; diagnosis ; Liver Failure ; diagnosis ; etiology ; Models, Biological ; Prognosis ; Proportional Hazards Models

Collagen Type I ; biosynthesis ; Hepatocyte Growth Factor ; biosynthesis ; Hepatocytes ; cytology ; metabolism ; physiology ; Liver Cirrhosis ; metabolism ; pathology ; physiopathology ; Liver Regeneration ; Receptors, Vitronectin ; biosynthesis

OBJECTIVETo analyze the clinical characteristics of plastic bronchitis associated with 2009 influenza A virus (H1N1) infection.

METHODA retrospective investigation of the clinical manifestation, bronchoscopy, and the histology of the cast, clinical course and outcome of 8 children with plastic bronchitis associated with influenza A virus (H1N1) infection during winter of 2009 and 2010 was performed.

RESULTAll 8 cases were boys, the range of age was 3 to 6 years. Five cases occurred in 2009 winter, accounting for 3.3% (5/150) of hospitalized children with influenza A (H1N1) infection; 3 cases occurred in 2010 winter, accounting for 15.8% (3/19) of hospitalized children with influenza A (H1N1) infection. Two patients had an underlying chronic disease, 1 had asthma, and the other had allergic rhinitis and atopic dermatitis. All the 8 cases had fever, cough and sputum; 2 had wheezing; 5 had respiratory distress. All 8 cases were diagnosed as influenza A virus (H1N1) infection complicated with pneumonia, of whom 5 patients had atelectasis, 2 had pneumothorax, 1 had pneumomediastinum, 1 had parapneumonic effusion, 2 patients were suspected of foreign body aspiration. Seven cases were admitted to an ICU, 5 patients developed respiratory failure, and 3 patients required mechanical ventilation. Flexible bronchoscopy and bronchial lavage was performed in all cases and showed bronchial cast. Histological examination of the bronchial cast revealed a fibrinous material containing large quantity of eosinophils, neutrophils, and lymphocytes in 7 patients, fibrinous material and necrotic material without inflammatory cells in 1 patient. After the bronchial cast was removed, all patients were improved greatly, no patients died.

CONCLUSIONPlastic bronchitis is a life-threatening complication associated with 2009 influenza A (H1N1) virus infection in children. In children with rapid and progressive respiratory distress with lung atelectasis or consolidation on chest radiograph, plastic bronchitis should be considered. Bronchoscopic extraction of casts should be carried out early.

Antiviral Agents ; administration & dosage ; therapeutic use ; Bronchitis ; complications ; diagnosis ; therapy ; virology ; Bronchoscopy ; Child ; Child, Preschool ; Foreign Bodies ; complications ; Glucocorticoids ; administration & dosage ; therapeutic use ; Humans ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; complications ; virology ; Intensive Care Units ; Male ; Pulmonary Atelectasis ; diagnosis ; therapy ; virology ; Rare Diseases ; Respiratory Insufficiency ; diagnosis ; therapy ; virology ; Retrospective Studies ; Tomography, X-Ray Computed ; Treatment Outcome

Objective To investigate the clinical characteristics of plastic bronchitis (PB) so as to improve the awareness of the disease.Methods Twenty-four children with PB were collected from Jul.2009 to Mar.2012 in Shenzhen Children's Hospital.The clinical manifestation,bronchoscopy,histology of the cast,clinical course and outcome were reviewed retrospectively.Results Of the 24 children with PB,18 cases were male,6 cases were female,and the range of age was 1 year and 2 months to 10 years and 3 months,with the median age of 3 years and 4 months.Three patients had an underlying chronic disease,1 case had asthma,1 case had hydronephrosis,and 1 case had ventricular septal defect repair before 1 year and 8 months.All the cases had fever,cough and sputum,while 10 cases had wheeze,and 5 cases had respiratory distress.All cases were diagnosed as pneumonia or severe pneumonia,of which 14 case had atelectasis,10 cases had parapneumonic effusion,5 cases suspected of foreign body inhalation,3 cases had pneumothorax,and 3 cases had mediastinal hernia.Fourteen cases were admitted to PICU,6 patients developed respiratory failure,and 9 patients required mechanical ventilation.Flexible bronchoscopy and bronchial lavage were performed in all cases and showed bronchial cast.Histological examination of the bronchial cast revealed that fibrinous material containing large quantity of eosinophils,neutrophils,and lymphocytes in 23 patients,and no inflammatory cells in 1 patient.After a bronchial cast was removed,all patients were improved greatly,and no patient dead.Conclusions Plastic bronchitis is a rare pediatric critical disease,which has high mortality.In children with rapid and progressive respiratory distress with lung atelectasis,pleural effusion or consolidation on chest radiograph,PB should be considered.Bronchial endoscopy is the most effective method for treatment of PB.

Objective To construct the recombinant varicella zoster virus (VZV)carrying 3xflag gene,3xflag was added to the VZV open reading frame 7 (ORF7)using GalK-based homologous recombination.Methods GalK and 3xflag gene fragments with 50 bp VZV ORF7 homologous arms were amplified by PCR.The obtained fragments were purified and transferred to the competent cells of VZV bacterial artificial chromosome (SW102-VZVWTBAC). The clones of VZV ORF7 with 3xflag (SW102-VZV ORF7-3xflag-BAC)were obtained by homologous recombination and selection from medium containing GalK and replaced GalK. The recombinant plasmids were extracted and transfected into ARPE-19 cells.The effect of VZV ORF7-3xflag on ARPE-19 cells was observed.Results The clones of VZV ORF7 with 3xflag (SW102-VZV ORF7-3xflag-BAC)were obtained.The virus patches with green fluorescence were observed three days after SW102-VZVWTBAC and SW102-VZV ORF7-3xflag-BAC were transfected into ARPE-19 cells.Western blot showed that ORF7 expression was effectively enhanced with 3xflag.Conclusion The recombinant VZV carrying 3xflag gene was obtained,which suggests that GalK-based homologous recombination is convenient,efficient and accurate in manipulating the gene virus of interest.