An exogenous avian leukosis virus (ALV) strain SDAU09C1 was isolated in DF-1 cells from one of 240 imported 1-day-old white meat-type grand parent breeder chicks. Inoculation of SDAU09C1 in ALV-free chickens induced antibody reactions specific to subgroup A or B. But gp85 amino acid sequence comparisons indicated that SDAU09C1 fell into subgroup A; it had homology of 88.8% - 90.3% to 6 reference strains of subgroup A, much higher compared to other subgroups including subgroup B. This is the first report for ALV of subgroup A isolated from imported breeders.

Objective To investigate the protective effect of the combination of glutamine (Gln) and ar-ginine (Arg) on intestinal barrier function in rats receiving fluorouracil (5-FU) chemotherapy.Methods Totally 40 male SD rats receiving 5-FU chemotherapy were equally randomized into four groups:enteral nutrition group,Gln group (enteral nutrition+Gln),Arg group (enteral nutrition+Arg),and Arg+Gln group (enteral nutri-tion+Arg+Gln).Observe the changes post chemotherapy such as the changes of body weight and urine lactulose/mannitol ratio before and after chemotherapy were recorded.On the 8 th post-therapy day,the blood endotoxin level in portal vein was measured,and lymph nodes and blood in portal vein were taken for bacterial culture;colon and jejunum specimens were also taken to measure the height of jejunum villus and the thickness of colon and jejunum.Results Body weights of Gln group,Arg group,and Arg+Gln group significantly increased after chemotherapy (P<0.05).The change of body weight was significantly lower in Arg+Gln group than in Gln group (P=0.002),while no such difference was found when compared with that in Arg group (P>0.05).Lactulose/manni-tol ratio in each group significantly increased after chemotherapy (P<0.05),and the change of lactulose/mannitol ratio was significantly higher in the enteral nutrition group than those in other groups (P=0.000);however,no such difference was found among other groups (P>0.05).The blood endotoxin level in portal vein was signifi-cantly higher in enteral nutrition group than in other groups (P=0.000);the endotoxin level was significantly lower in Gln group than in Arg group (P=O.035) and Arg+Gln group (P=0.000);however,no such differ-ence was found between Arg group and Arg+Gln group (P=0.109).The height of jejunum villus and the thick-ness of jejunum were significantly lower in enteral nutrition group than those in the other groups (P=O.000);the thickness of colon was significantly lower in enteral nutrition group than those in Arg group and Arg+Gln group (P=0.000);however,no such difference was found when compared with Gln group (P=0.058).The thickness of colon (P=0.040) and jejunum (P=0.010) was significantly higher in Gln group than that in Arg group;but there was no significant difference in term of the height of jejunum villus (P=0.286).Compared with Arg+Gln group,the thickness of jejunum in Gln group was significantly higher,but there was no significant difference in terms of the height of jejunum villus (P=0.286) or the thickness of jejunum (P=0.190).The thickness of co-lon was significantly lower in Arg group than in Arg+Gln group (P=0.010),while no such significant difference was found in terms of the height of jejunum villus (P=0.803) or the thickness of colon (P=0.059) when com-pared with Arg+Gln group.The bacterial culture results were not significantly different among all groups.Conclu-sions The combination of Arg and Gln has protective effects on intestinal barrier in rats receiving 5-FU chemother-apy.The protective effect of Gln is superior to that of Arg.No synergistic effect exists between Arg and Gln.

BACKGROUND: Endoscopic transnasal lacrimal duct reconstruction by grafting of autogenous tissue is a novel method for treatment of severe lacrimal duct obstruction and it needs detailed anatomical data for surgery.OBJECTIVE: To study the applied microsurgical anatomy of lacrimal duct and to provide anatomical evidence for endoscopic transnasal lacrimal duct reconstruction by grafting of autoganous tissue.DESIGN, TIME AND SETTING: This study was performed at the laboratory of the Department of Ophthalmology, Armed Police General Hospital from July 2006 to June 2007.MATERIALS: Twenty 10% formaldehyde-treated adult cadaveric heads, 14 males and 6 females, comprising 40 lacrimal ducts were included in this study.METHODS: The cadaveric heads were split on the level of the line between the superior border of the superciliary arch and the site 10 mm higher than occipital tuberosity. After removal of brain tissue,the heads were decalcified for approximate 1 week with 10%nitric acid. This promised non-alteration of morphological structure and facilitation for surgical cutting. Following dissection of facial cranium in the median sagittal plane, the nasal septum was excised to expose the lateral wall of the nasal cavity.MAIN OUTCOME MEASURES: The anteroposterior diameter and depth of lacrimal fossa; at middle third level, the thickness of lacrimal fossa at the anterior lacrimal crest, vertical middle line, and posterior lacrimal crest; the cross section area of nasolacrimal canal upper opening, middle part, and lower opening; horizontal distance, 30° oblique distance, and 45°oblique distance from lacrimal caruncie to nasal cavity; distance from lacrimal caruncle to nasolacrimal canal upper opening; and the included angle between lacrimal caruncle-nasolacrimal canal upper opening line and Aeby's plane.RESULTS: The length, anteroposterior diameter, and depth of lacrimal fossa were (17.85±1.72) mm, (6.74+1.28) mm, and (3.09+0.78) mm, respectively. At middle third level, the thickness of lacrimal fossa at the anterior lacrimal crest,perpendicular bisector, and posterior lacrimal crest was (4.03±0.89) mm, (0.61±0.36) mm, and (0.63±0.24) mm, respectively.Anterior lacrimal crest was significantly thicker than vertical middle line and posterior lacrimal crest (P ＞ 0.05). Horizontal distance, 30°oblique distance, and 45° oblique distance from lacrimal caruncle to nasal cavity was (17.23±0.70) mm,(14.51±1.72) mm, and (17.34±2.38) mm, accordingly, with a difference which was not significant (P ＞ 0.05). The distance from lacrimal caruncle to lateral wall middle point of nasolacrimal duct superior opening was (11.86±1.84) mm, and the included angle between lacrimal caruncle-lateral wall middle point of nasolacrimal duct superior opening line and Aeby's plane averaged (49.9±1.8)°.CONCLUSION: The distances from lacrimal caruncle to nasal cavity and lacrimal sac and the included angles between lacrimal caruncle-nasolacrimal canal upper opening line and Aeby's plane provide guidance significance for selection of bony opening position on the lateral wall of nasal cavity and determinations of tunnel oblique angle and autogenous tissue length. Creation of bony tunnel should start from the middle or posterior middle part of lacrimal fossa and then extend towards anterior inferior region with an optimal downward oblique angle of 45°. The length of autogenous tissue used for lacrimal duct reconstruction should exceed 21.22 mm.

In order to observe the effect of the immune and weight of chickens after use the attenuated vaccine with low dose of chicken infectious anemia virus (CIAV). In this study, the effects of low dose of CIAV on the weight of SPF chickens and NDV antibody production were observed by simulated experiments. The results showed that 10 EID50 and 5 EID50 CIAV per plume attenuated NDV vaccines were used to cause the weight loss of SPF chickens. Compared with the use of the non contaminated vaccine group, it has significant difference. And NDV antibody levels compared with the use of the non contaminated groups also decreased after use the vaccine with two doses of CIAV contaminated. It has significant difference. A certain proportion of CIAV antibody positive was detected at the beginning of the second week after use the NDV vaccine with two doses of CIAV contaminated. The detection of a high proportion of CIAV nucleic acid was detected in the first week after the use of a contaminated vaccine. The results of the study demonstrate the effects of CIAV pollution on the production and immune function of SPF chickens, and it is suggested that increasing the detection of viral nucleic acid can help save time and improve the detection rate in the detection of exogenous virus contamination by SPF chicken test method.
Animals ; Antibodies, Viral ; immunology ; Chicken anemia virus ; genetics ; immunology ; physiology ; Chickens ; Circoviridae Infections ; immunology ; veterinary ; virology ; Poultry Diseases ; immunology ; virology ; Specific Pathogen-Free Organisms ; Vaccines, Attenuated ; administration & dosage ; genetics ; immunology

To understand epidemiological characteristics of Marek's disease virus (MDV) prevalent in china currently,3 Marek's disease (MD) strains were isolated and identified from white feather meat chickens vaccined with MDV CVI988 or 814 through necropsy,histopathological observation,virus isolation and IFA detection,named SDAU1501,SDAU1502 and SDAU1503,respectively.vIL8,pp38,MEQ gene of the three strains of MDV were amplified using PCR,and compared with reference strains.The homology between SDAU1501 and SDAU1502 and virulent strains was above 97％,suggesting some features of virulent strains;while meq gene of SDAU1503 lost P amino acid at the 194 th site as that in CVI988,But the distinctive 177 nucleotide insertion mutations was not existed,predicting that it may be a attenuated vaccine strain.New variations of MDV continued and different types of variants emerged,therefore,prevalence and genetic monitoring of MD should be proceeded;meanwhile,more attentions should be given to MDV vaccine development.

We used a meq-deleted attenuated MDV-I strain GX0101Δmeq as a vector to construct a recombinant virus expressing the exogenous gene NDV-F. The ORF of exogenous gene NDV-F was inserted into the eukaryotic expression vector pcDNA3.1(-). Then, the expression cassette of NDV-F which contains the CMV promoter was amplified. Simultaneously, we amplified the selected gene Kan+ expression cassette and inserted them into the PMD18-T vector. Tandem expression cassettes were amplified using primers containing the 50-bp homologous arm of MDV-US2. The PCR product was electroporated into EL250 host bacteria containing GX0101Δmeq. Then, the Kan+ expression cassette was deleted from the recombinant virus genome using 1% arabinose. The plasmid of the positive clone which the Kan+ expression cassette was deleted was extracted and transfected into CEFs to rescue the recombinant virus. The recombinant virus was injected into chickens to observe its growth and replication. The recombinant virus rMDV-F containing the exogenous gene NDV-F was rescued successfully. The recombinant virus could duplicate and express well in CEFs, and grow and replicate well in chickens. Using GX0101Δmeq as a vector, combined with a recombinant system of Red E/T and FLP/FRT, we constructed a recombinant virus that expressed the exogenous gene NDV-F. This study could lay the foundation for further study of recombinant viruses.
Animals ; Cell Line ; Chickens ; virology ; DNA, Recombinant ; genetics ; Gene Expression ; Genetic Engineering ; Genetic Vectors ; genetics ; Mardivirus ; genetics ; physiology ; Plasmids ; genetics ; Viral Proteins ; genetics ; Virus Replication

The genomic diversity of Avian leukosis virus subgroup J (ALV-J) was investigated in an experimentally infected chicken. ALV-J variants in tissues from four different organs of the same bird were re-isolated in DF-1 cells, and their gp85 gene was amplified and cloned. Ten clones from each organ were sequenced and compared with the original inoculum strain, NX0101. The minimum homology of each organ ranged from 96.7 to 97.6%, and the lowest homology between organs was only 94.9%, which was much lower than the 99.1% homology of inoculum NX0101, indicating high diversity of ALV-J, even within the same bird. The gp85 mutations from the left kidney, which contained tumors, and the right kidney, which was tumor-free, had higher non-synonymous to synonymous mutation ratios than those in the tumor-bearing liver and lungs. Additionally, the mutational sites of gp85 gene in the kidney were similar, and they differed from those in the liver and lung, implying that organ- or tissue-specific selective pressure had a greater influence on the evolution of ALV-J diversity. These results suggest that more ALV-J clones from different organs and tissues should be sequenced and compared to better understand viral evolution and molecular epidemiology in the field.
Animals ; Avian Leukosis Virus* ; Avian Leukosis* ; Birds ; Chickens* ; Clone Cells ; Kidney ; Liver ; Lung ; Molecular Epidemiology ; Silent Mutation

OBJECTIVETo explore the effects of acupuncture on the expression of protein kinase B (PKB/AKT) in lung tissues of asthma rats.

METHODSForty SPF male SD rats were randomly divided into a blank group, a model group, an acupuncture group and a blocker group, 10 rats in each one. The rat model of asthma was established by egg albumin stimulation in the model group, acupuncture group and blocker group. Since the establishment of rat model, the rats in the acupuncture group were treated with acupuncture at "Dazhui" (GV 14), "Feishu" (BL 13) and "Fengmen" (BL 12) before atomization; the rats in the blocker group were treated with intervention of blocker LY294002, once every two days, for 7 times. There was no treatment in the blank group and model group. HE staining was applied to observe the morphologic changes of lung tissues; the immunohistochemical method was applied to test the protein expression of AKT in lung tissue.

RESULTSHE staining indicated the infiltration and aggregation of a variety of inflammatory cells around airways, as well as bronchial smooth muscle spasm and confined lumen in the model group; in the acupuncture group and blocker group the inflammatory cells were less and confined lumen was relieved. Compared with the blank group, the protein expression of AKT was higher in the model group (<0.05); compared with the model group, the protein expression of AKT in the acupuncture group and blocker group was reduced (both<0.05); the differences between the acupuncture group and blank group, blocker group were not significant (both>0.05).

CONCLUSIONSAcupuncture could reduce the protein expression of AKT in lung tissue in asthma rats, leading to relieved inflammation reaction and airway remodeling.

Flagella are the main motility structure of Clostridioides difficile that affects the adhesion, colonization, and virulence of C. difficile in the human gastrointestinal tract. The FliL protein is a single transmembrane protein bound to the flagellar matrix. This study aimed to investigate the effect of the FliL encoding gene flagellar basal body-associated FliL family protein (fliL) on the phenotype of C. difficile. The fliL gene deletion mutant (ΔfliL) and its corresponding complementary strains (: : fliL) were constructed using allele-coupled exchange (ACE) and the standard molecular clone method. The differences in physiological properties such as growth profile, antibiotic sensitivity, pH resistance, motility, and spore production ability between the mutant and wild-type strains (CD630) were investigated. The ΔfliL mutant and the : : fliL complementary strain were successfully constructed. After comparing the phenotypes of strains CD630, ΔfliL, and : : fliL, the results showed that the growth rate and maximum biomass of ΔfliL mutant decreased than that of CD630. The ΔfliL mutant showed increased sensitivity to amoxicillin, ampicillin, and norfloxacin. Its sensitivity to kanamycin and tetracycline antibiotics decreased, and the antibiotic sensitivity partially returned to the level of CD630 strain in the : : fliL strain. Moreover, the motility was significantly reduced in the ΔfliL mutant. Interestingly, the motility of the : : fliL strain significantly increased even when compared to that of the CD630 strain. Furthermore, the pH tolerance of the ΔfliL mutant significantly increased or decreased at pH 5 or 9, respectively. Finally, the sporulation ability of ΔfliL mutant reduced considerably compared to the CD630 strain and recovered in the : : fliL strain. We conclude that the deletion of the fliL gene significantly reduced the swimming motility of C. difficile, suggesting that the fliL gene is essential for the motility of C. difficile. The fliL gene deletion significantly reduced spore production, cell growth rate, tolerance to different antibiotics, acidity, and alkalinity environments of C. difficile. These physiological characteristics are closely related to the survival advantage in the host intestine, which is correlated with its pathogenicity. Thus, we suggested that the function of the fliL gene is closely related to its motility, colonization, environmental tolerance, and spore production ability, which consequently affects the pathogenicity of C. difficile.
Humans ; Clostridioides/metabolism* ; Clostridioides difficile/metabolism* ; Bacterial Proteins/metabolism* ; Virulence ; Anti-Bacterial Agents/metabolism*