Avian influenza viruses (AIV) can cause serious economic losses and threaten to human health. The laboratory methods for rapid and accurate detecting AIV ensure timely implementation of intervention strategies so that it plays a key role in avian influenza prevention and control. The laboratory technologies for detecting AIV are topic subject and developed quickly those days. This paper reviews the advances of laboratory diagnosis technologies for AIV at 3 aspects of virus isolation, immunoassay and molecular diagnostics.

This article aimed to study the antigenicity of nucleocapsid proteins (NPs) in six pathogenic phleboviruses and to provide theoretical evidence for the development of serological diagnostic reagents. NPs of six pathogenic phleboviruses were expressed and purified using a prokaryotic expression system and rabbits were immunized with individual recombinant NPs. Cross-reactions among NPs and rabbit sera were determined by both indirect ELISA and Western blotting analyses, and the sera titer was determined by indirect ELISA. Furthermore, sera from SFTS patients were also detected by each recombinant NP as a coating antigen using indirect ELISA. The cross-reactions and the sera titer were subsequently determined. Both the concentration and purity of recombinant NPs of six pathogenic phleboviruses met the standards for immunization and detection. The results of indirect ELISA and Western blotting showed that each anti-phlebovirus NP rabbit immune serum had potential serological cross-reactivity with the other five virus NP antigens. Furthermore, the sera from SFTS patients also had cross-reactivity with the other five NP antigens to a certain extent. Our preliminary study evaluated the antigenicity and immune reactivity of six pathogenic phleboviruses NPs and laid the foundation for the development of diagnostic reagents.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; genetics ; immunology ; Cross Reactions ; Humans ; Nucleocapsid Proteins ; genetics ; immunology ; Phlebotomus Fever ; diagnosis ; immunology ; virology ; Phlebovirus ; classification ; genetics ; immunology ; isolation & purification ; Rabbits

To explore the expression potential of heterogeneous genes using the backbone of infectious bronchitis virus (IBV) Beaudette strain, the ectodomain region of the Spike gene (1,302 bp) of IBV H120 strain was amplified by RT-PCR and replaced into the corresponding location of the IBV Beaudette strain full-length cDNA. This recombinant was designated as BeauR-H120(S1). BeauR-H120(S1) was directly used as the DNA template for the transcription of viral genomic RNA in vitro. Then, the transcription product was transfected into Vero cells by electroporation. At 48 h post-transfection, the transfected Vero cells were harvested, and passaging continued. A syncytium was not observed until the recombinant virus had passed through four passages. The presence of rBeau-H120(S1) was verified by the detection of the replaced ectodomain region of the H120 Spike gene using RT-PCR. Western blot analysis of rBeau-H120 (S1)-infected Vero cell lysates demonstrated that the nucleocapsid (N) protein was expressed, which implied that rBeau-H120(S1) could propagate in Vero cells. The TCIDs0 and EIDs0 data demonstrated that the titer levels of rBeau-H120(S1) reached 10(590+/-0.22)TCID50/mL and 10(6.13+/-0.23)EID50/mL in Vero cells and 9-day-old SPF chicken embryos, respectively. Protection studies showed that the percentage of antibody-positive chickens, which were vaccinated with rBeau-H120(S1) at 7 days after hatching, rose to 90% at 21 days post-inoculation. Inoculation provided an 85% rate of immune protection against a challenge of the virulent IBV M41 strain (103EID50/chicken). This recombinant virus constructed using reverse genetic techniques could be further developed as a novel genetic engineering vaccine against infectious bronchitis.
Animals ; Cercopithecus aethiops ; Chick Embryo ; Chickens ; Coronavirus Infections ; veterinary ; virology ; Infectious bronchitis virus ; chemistry ; genetics ; growth & development ; metabolism ; Poultry Diseases ; virology ; Protein Structure, Tertiary ; Spike Glycoprotein, Coronavirus ; chemistry ; genetics ; metabolism ; Transfection ; Vero Cells

OBJECTIVETo explore the relationship between expression of Fas, bcl-2 and apoptosis of cardiomyocytes in myocardial ischemia in rats.

METHODSMyocardial ischemia was experimentally induced by ligating the left coronary artery. The rate were killed from 10 minutes to 7 days after the operation. Apoptotic myocardial cells were detected by TUNEL method. The expression of Fas and bcl-2 was studied by ABC immunohistochemistry.

RESULTSCardiomyocytic apoptosis appeared from 3 to 36 hours after ischemia. This phenomenon however could not be detected by TUNEL method 7 days after ischemia. The expression of Fas could be detected by ABC immunohistochemistry from 3 hours to 7 days after ischemia, and the expression of bcl-2 from 10 minutes to 7 days. Cardiomyocytic apoptosis and Fas / bcl-2 expression appeared in different regions of myocardium: apoptosis in the ischemic regions, while Fas and bcl-2 expression in regions without obvious ischemia.

CONCLUSIONIn rats, Fas and bcl-2 may not directly regulate apoptosis of cardiomyocytes in case of myocardial ischemia.

Animals ; Apoptosis ; Male ; Myocardial Infarction ; metabolism ; pathology ; Myocytes, Cardiac ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; fas Receptor ; metabolism

Objective To explore the effects and safety of hemoperfusion(HP) therapy on tetramine poisoning in infants.Methods Thirty-five infants with tetramine poisoning were divided into two groups: HP group(n=18) and non HP group(n=17).The changes of blood tetramine concentration and clinical symptom improving of both groups after the treatment were observed together with the adverse effects of HP group.Results The average blood tetramine concentration of HP group was higher than that of non HP group(342.2?333.4 vs 117.9?50.8 ?g/L,P

OBJECTIVETo study the incidence of human cytomegalovirus (CMV) and human herpesvirus 6 (HHV-6) infection in pediatric patients with hemopoietic stem cell transplantation (HSCT), and to explore the relationship between CMV and HHV-6 infection in pediatric patients with HSCT.

METHODSPediatric patients with HSCT in hemotology center of Beijing Children's Hospital were enrolled into this study from June 2007 to October 2009. Peripheral blood were collected every week after HSCT, and Fluorescent quantitation PCR and conventional PCR were used to detect CMV DNA load in serum and HHV-6 DNA in peripheral blood respectively. Genetic typing was conducted on HHV-6.

RESULTSFifty two pediatric patients with HSCT were enrolled into this study, and six hundreds and thirty six specimens were collected totally. CMV DNA was detected in fifty two specimens from twenty cases. The median time was 56 days after HSCT. The incidence of CMV infection was 38.5% (20/52) in all HSCT patients and 47.6% (20/42) in allogene HSCT patients. The incidence of late CMV infection was 22.2% (6/27) in allogene HSCT. Three patients died of CMV infection,and two died of CMV interstitial pneumonia. HHV-6 DNA was detected in thirty three specimens from fourteen cases. The median time was 23 days after HSCT. The incidence of HHV-6 infection was 26.9% (14/52)in all HSCT patients and 31% (13/42) in allogene HSCT patients. The genotype of HHV6 was all type B. HHV-6 DNA was positive in six of twenty cases with CMV infection. The incidence of co-infection was 30% (6/20).

CONCLUSIONSThere was a substantial incidence of CMV and HHV6 infection after HSCT. The relationship between earlier HHV6 infection and later CMV infection in pediatric patients with HSCT need further study.

Adolescent ; Child ; Child, Preschool ; Cytomegalovirus ; genetics ; Cytomegalovirus Infections ; etiology ; immunology ; Female ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Herpesviridae Infections ; etiology ; immunology ; Herpesvirus 6, Human ; genetics ; Humans ; Infant ; Male ; Molecular Typing ; methods

AIMTo explore the influence of hyperbaric oxygen (HBO) preconditioning on anoxic resistance and anti-weariness at high altitude.

METHODS(1) SOD, MDA, NO, NOS, BLA and BUN of 20 youths living at 3 700 m altitude for half year were tested, then they were divided into group A (n=10, received HBO pretreatment twice) and group B (n=10, received HBO pretreatment 5 times) randomly. They were asked to pedal the EMG-bicycle-ergometer at the second and eighth day, and then the same items were tested. (2) 29 youth who would go to Astronomical Spot (5200 m) were randomly divided into group HBO (n=11, received HBO pretreatment once per day for 2 days at Yecheng (1400 m)) and comparison group (n=10). When they reached I Astronomical Spot, thematic biochemical index were investigated. (3) When 20 youth reached Thirty Milepost Barracks (3700 m) at the second day in their way to Immortal Gulf (5380 m) from Yecheng were randomly divided into group HBO (n=10, received HBO pretreatment once per day for 3 days) and comparison group (n=10). When they reached Immortal Gulf, the thematic biochemical index were investigated.

RESULTS(1) SOD, NO, NOS were increased and BLA, BUN, MDA were decrease in group A compared with that in group B until the eighth day, there was significant difference (P < 0.01). (2) SOD, NO, NOS were increased and BLA, BUN, MDA were decrease in group HBO compared with that in comparison that in group, there was significant difference between groups (P < 0.01, or P < 0.05).

CONCLUSIONHBO could enhance the activity of anti-oxidase and the cleared ability of lactic acid, and the effect of anti-weariness could last for 8 days.

Altitude ; Altitude Sickness ; prevention & control ; Fatigue ; prevention & control ; Humans ; Hyperbaric Oxygenation ; methods ; Hypoxia ; physiopathology ; Ischemic Preconditioning ; methods ; Male ; Oxidative Stress ; drug effects ; physiology ; Superoxide Dismutase ; metabolism ; Young Adult

OBJECTIVETo study the correlation of polymorphisms of CYP1A1 MSPI and glutathiones S-transferase (GST-M1) independently and in combination with the risk of lung cancer.

METHODSA case control study which included 91 cases of lung cancer and 138 controls collected from the First Affiliated Hospital of Sun Yat-sen University of Medical Sciences, Guangzhou Tumor Hospital and The Red Cross Hospital of Guangzhou or conmunity area. All subjects were investigated with a uniform questionnaire. Blood samples were collected from all cases and controls for detecting CYP1A1 MSPI and GST-M1 polymorphisms which were analyzed by PCR and RFLP.

RESULTSIt showed that there was no significant difference in frequencies of this genotypes of CYP1A1 MSPI between the two groups. The frequency of GST-M1 null (0/0) genotype was higher in the case group than in the control group, with an OR of 1.38 (95% CI 0.81 - 2.38), but there was no statistical significance. However, combination of several genotypes was strongly associated with lung cancer. There was a synergistic interaction between the m2m2 genotype of CYP1A1 MSPI and GST-M1 (0/0) genotype, with an OR of 2.47 (95% CI 1.03 - 5.90).

CONCLUSIONThe combination of two genetic polymorphisms significantly increases the risk of lung cancer.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Cytochrome P-450 CYP1A1 ; genetics ; Female ; Genotype ; Glutathione Transferase ; genetics ; Humans ; Lung Neoplasms ; etiology ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; Risk ; Smoking ; adverse effects

OBJECTIVETo review retrospectively the experience of surgical repair of sinus of valsalva aneurysm (SVA) in 70 patients.

METHODSBetween September, 1988 and October, 2003, Seventy patients with SVA underwent surgical repair by the aid of general anesthesia and cardiopulmonary bypass, comprised 1.4% (70/4960) of all open-heart operation. Forty-five were male and 25 female. Age ranged from 3 to 69 years old [mean (29 +/- 15) years]. The aneurysms ruptured into the right ventricle in 46 patients, right atrium in 23 and left ventricle in 1 respectively. The aneurysms originated from right and noncoronary sinus in 61 patients (87%) and 9 patients (13%) respectively. The most common associated cardiovascular lesions were ventricular septal defect (VSD, n = 34) and aortic valve incompetence (n = 21). Repairs were achieved through an incision in right atriotomy, right ventriculotomy or aortotomy only or both aortotomy and right atriotomy (or right ventriculotomy). The defects in the sinus of valsalva was repaired with either direct sutures (n = 43) or a patch (n = 27). The aortic valve was replaced in 6 patients.

RESULTSThere were no deaths in early time after repair. Postoperative hospital stay was 8 approximately 33 days [mean +/- standard deviation, (14.3 +/- 6.4) days] before 1997 and 6 approximately 15 days [mean +/- standard deviation, (9.1 +/- 2.6) days] after 1997 respectively. Complications included infection (n = 4), hemorrhage (n = 4), pneumothorax (n = 1), arrhythmia (n = 4) and residual shunt (n = 1) of VSD. Fifty-three (76%) patients (2 months approximately 13 years) were followed-up [mean +/- standard deviation, (6.6 +/- 3.8) years]. All patients survived except that one died of rupture of dissecting aortic aneurysm 7 years after operation.

CONCLUSIONSThe ruptured sinus of valsalva aneurysm and unruptured sinus of valsalva aneurysm with ventricle septal defect or(and) aortic valve regurgitation should be repaired surgically as soon as the diagnosis was confirmed. Long-term results are associated with preoperative aortic valve regurgitation.

Adolescent ; Adult ; Aged ; Aortic Aneurysm ; complications ; surgery ; Aortic Rupture ; surgery ; Aortic Valve Insufficiency ; complications ; surgery ; Child ; Child, Preschool ; Female ; Heart Septal Defects, Ventricular ; complications ; surgery ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Sinus of Valsalva ; surgery ; Treatment Outcome

OBJECTIVETo screen and obtain the validate immunogenic membrane antigens in pancreatic cancer.

METHODSPancreatic cancer cell line SW1990 membrane protein was extracted and separated by two-dimensional gel electrophoresis (2-DE). One of the three parallel 2-DE gels underwent Coomassie blue staining while the other two underwent immunoblot. Serum IgG was purified from clinically collected sera of 66 pancreatic cancer patients and 24 chronic pancreatitis patients and used as the primary antibody of the immunoblot. Positive dots of immunoblot were identified by MALDI-TOF mass spectrometry and PMF matching. The candidate membrane antigens were further validated respectively in cell lines and tissues by RT-PCR, real-time PCR, Western blot, and their different expression level of gene and protein between pancreatic cancer cell line and normal pancreatic tissue were compared studied.

RESULTSThe immunoblot of SW1990 membrane protein with serum IgG from cancer patients showed nine positive dots which were not the same as those from immunoblot with serum IgG from chronic pancreatitis patients. One talent dot was identified with MALDI and PMF as VDAC2. RT-PCR and real-time PCR showed that the gene of VDAC2 was expressed in the pancreatic cancer cell line. Western blot showed that the expression of protein level of VDAC2 in the pancreatic cancer cell line was obviously higher than in normal pancreatic tissue.

CONCLUSIONSVDAC2 might be the candidate immunogenic membrane antigens of pancreatic cancer, and its gene is all expressed in the pancreatic cancer cell line SW1990, AsPc and P3. The protein level of VDAC2 is significantly overexpressed in pancreatic cancer cell line than in normal pancreatic tissue.

Adult ; Aged ; Antigens, Neoplasm ; isolation & purification ; Cell Line, Tumor ; Female ; Humans ; Male ; Membrane Proteins ; immunology ; isolation & purification ; Middle Aged ; Pancreatic Neoplasms ; immunology ; Proteomics ; Voltage-Dependent Anion Channel 2 ; metabolism ; Young Adult