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

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

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

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

BACKGROUNDIt is a surgical dilemma when patients present with both severe heart disease and neoplasms. The best surgical treatment remains controversial. This study aimed to analyze the early and long-term results of simultaneous surgical treatment of severe heart disease and neoplasms.

METHODSWe reviewed the clinical records of 15 patients who underwent simultaneous neoplastic resection and cardiac surgery between September 2006 and January 2011. There were 5 male and 10 female patients. The mean age was (59.2 ± 12.5) years and the mean left ventricular ejection fraction was (57.4 ± 11.0)%. All patients were followed up completely for a period of 12 to 51 months (mean, (33.1 ± 11.2) months).

RESULTSFifteen patients underwent simultaneous cardiac surgery and neoplastic resection. Cardiac procedures consisted of off pump coronary artery bypass grafting (n = 7), aortic valve replacement (n = 3), mitral valve replacement (n = 3), mitral valve replacement with coronary artery bypass grafting (n = 1) and left atrial myxoma resection (n = 1). Neoplastic resection consisted of lung cancer resection (n = 5), colonic cancer resection (n = 3), gallbladder resection (n = 1), colonic cancer resection with gallbladder resection (n = 1), hysterectomy (n = 2), hysterectomy with bilateral salpingo-oophorectomy (n = 2) and left ovariectomy (n = 1). Pathological examination confirmed malignant disease in 10 patients and benign disease in 5 patients. There were no perioperative myocardial infarctions, stroke, pericardial tamponade, renal failure or hospital deaths. The most frequent complications were atrial fibrillation (33.3%), pneumonia (26.7%), low cardiac output syndrome (6.7%) and delayed healing of surgical wounds (6.7%). There was 1 late death 42 months after surgery for recurrent malignant disease. At 1 and 3 years, survival rates were 100% (Kaplan-Meier method).

CONCLUSIONSSimultaneous cardiac surgery and neoplastic resection was not associated with increased early or late morbidity or mortality. Cardiopulmonary bypass does not appear to adversely affect survival in patients with malignant disease. The long-term survival was determined by tumor stage.

Adult ; Aged ; Colonic Neoplasms ; surgery ; Female ; Heart Diseases ; surgery ; Humans ; Hysterectomy ; adverse effects ; Lung Neoplasms ; surgery ; Male ; Middle Aged ; Ovariectomy ; adverse effects ; Thoracic Surgery ; statistics & numerical data ; 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

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

AIMTo investigate the effects of glycyrrhiza decoction on migrating myoelectric complex (MMC) and gastrointestinal hormone in small intestine in rats.

METHODSWe observed MMC cycle,phase Ill duration,fast wave numbers of phase III of MMC in one minute, fast wave numbers of one cluster in phase III of MMC of small intestine of glycyrrhiza group and control group rats with electrophysiology method, and immunohistochemistry to examine relative content of serotonin (5-HT), substance p(SP) and vasoactive intestinal polypeptide (VIP) in small intestinal chromophil (EC) and myenteric nerve plexus in small intestine of control group and glycyrrhiza group rats.

RESULTSCompared glycyrrhiza group with control group,we found that glycyrrhiza was able to decrease fast wave numbers in one minute and fast wave numbers in one cluster in phase III of MMC of small intestine (P < 0.05), and evidently extend small intestinal cycle of MMC (P < 0.05), it also shortened the phase III III duration (P < 0.05) or made the phase III of MMC absent. Compared glycyrrhiza group with control group it was indicated that content of 5-HT in small intestinal mucous membrane and myenteric nerve plexus was evidently decreased (P < 0.05), and content of SP in myenteric nerve plexus of small intestine of rats was evidently decreased (P < 0.05), and content of VIP in small intestine of rats was evidently increased (P < 0.05).

CONCLUSIONGlycyrrhiza is able to inhibit small intestinal motility, this inhibition is related with the amount of 5-HT, SP, VIP secreted by small intestinal mucous membrane of rats.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Electromyography ; Gastrointestinal Motility ; drug effects ; physiology ; Glycyrrhiza ; Intestine, Small ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley ; Serotonin ; analysis ; Substance P ; analysis ; Vasoactive Intestinal Peptide ; analysis

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