Annexin A2 gene interacting with viral matrix protein to promote bovine ephemeral fever virus release

Bovine ephemeral fever virus (BEFV) causes bovine ephemeral fever, which can produce considerable economic damage to the cattle industry. However, there is limited experimental evidence regarding the underlying mechanisms of BEFV. Annexin A2 (AnxA2) is a calcium and lipid-conjugated protein that binds phospholipids and the cytoskeleton in a Ca 2+ -dependent manner, and it participates in various cellular functions, including vesicular trafficking, organization of membrane domains, and virus proliferation. The role of the AnxA2 gene during virus infection has not yet been reported. In this study, we observed that AnxA2 gene expression was up-regulated in BHK-21 cells infected with the virus.Additionally, overexpression of the AnxA2 gene promoted the release of mature virus particles, whereas BEFV replication was remarkably inhibited after reducing AnxA2 gene expression by using the small interfering RNA (siRNA). For viral proteins, overexpression of the Matrix (M) gene promotes the release of mature virus particles. Moreover, the AnxA2 protein interaction with the M protein of BEFV was confirmed by GST pull-down and coimmunoprecipitation assays. Experimental results indicate that the C-terminal domain (268– 334 aa) of AxnA2 contributes to this interaction. An additional mechanistic study showed that AnxA2 protein interacts with M protein and mediates the localization of the M protein at the plasma membrane. Furthermore, the absence of the AnxA2-V domain could attenuate the effect of AnxA2 on BEFV replication. These findings can contribute to elucidating the regulation of BEFV replication and may have implications for antiviral strategy development.

Rapid visual detection of Mycobacterium avium subsp. paratuberculosis by recombinase polymerase amplification combined with a lateral flow dipstick

Paratuberculosis (Johne's disease) is a chronic debilitating disease of domestic and wild ruminants. However, widespread point-of-care testing is infrequent due to the lack of a robust method. The isothermal recombinase polymerase amplification (RPA) technique has applied for rapid diagnosis. Herein, RPA combined with a lateral flow dipstick (LFD) assay was developed to estimate DNA from Mycobacterium avium subsp. paratuberculosis. First, analytical specificity and sensitivity of the RPA-nfo primer and probe sets were assessed. The assay successfully detected M. paratuberculosis DNA in 30 min at 39℃ with a detection limit of up to eight copies per reaction, which was equivalent to that of the real-time quantitative polymerase chain reaction (qPCR) assay. The assay was specific, as it did not amplify genomes from five other Mycobacterium spp. or five pathogenic enteric bacteria. Six hundred-twelve clinical samples (320 fecal and 292 serum) were assessed by RPA-LFD, qPCR, and enzyme-linked immunosorbent assay, respectively. The RPA-LFD assay yielded 100% sensitivity, 97.63% specificity, and 98.44% concordance rate with the qPCR results. This is the first report utilizing an RPA-LFD assay to visualize and rapidly detect M. paratuberculosis. Our results show this assay should be a useful method for the diagnosis of paratuberculosis in resource-constrained settings.

Establishment and Preliminary Application of the SYBR Green I Real-time PCR Assay for Detection of the Bovine Enterovirus / 病毒学报

The bovine enterovirus (BEV) is a pathogen found the digestive tracts of cattle. Recently, the BEV was discovered in cattle in a province in China. A rapid and effective detection method for the BEV is essential. An assay was carried out using two specific primers designed to amplify a highly conserved sequence of the 3D gene. A recombinant plasmid containing the target gene 3D was constructed as a standard control. The limit of detection of the reaction was 7.13 x 10(1) plasmid copies/μL of initial templates, which was tenfold more sensitive than the conventional reverse-transcription-polymerase chain reaction (RT-PCR). Moreover, the assay was highly specific because all negative controls and other viruses of clinical relevance did not develop positive results. Assay performance on field samples was evaluated on 44 (41 diarrhea and 3 aerosol) samples and compared with the conventional RT-PCR assay. Sixteen diarrhea samples were positive (16/41, 39. 02%) and 3 aerosol samples were positive (3/3, 100%). Preliminary results for clinical detection showed that the SYBR Green I real-time PCR assay was highly sensitive, specific and reproducible. The robustness and high-throughput performance of the developed assay make it a powerful tool in diagnostic applications for epidemics and in BEV research.

The effect of endoscopic sinus surgery on pulmonary function of chronic rhinosinusitis patients with asthma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To explore the effection of the pulmonary function of patients of chronic rhinosinusitis (CRS) with asthma which treated with endoscopic sinus surgery (ESS) based comprehensive treatment.@*METHOD@#There were 50 cases of chronic rhinosinusitis with asthma whom met the study criteria. 35 cases enrolled in the tri al group, which treated with endoscopic sinus surgery, and routine perioperative tratment. Another 15 cases as control group which underwent conservative treatment. Both groups underwent the rule treatment of asthma. The main monitoring indexes, which included visual analogue scale (VAS) score, endoscopic Lund-Kennedy score, control of asthma symptoms, the pulmonary function which involved forced expiratory volume in first second (FEV1), forced vital capacity (FVC), the ratio of forced expiratory volume in first second and forced vital capacity (FEV1/FVC) and peak expiratory flow (PEF), were measured in the patients of each groups before surgery, follow-up for 1 year and 3-year.@*RESULT@#Our study found that the VAS score of CRS with asthma was significantly negatively correlated with FEV1 and PEF (P < 0.05), endoscopic Lund-Kennedy score was significantly negatively correlated with PEF (P < 0.05); After the trial group underwent ESS based comprehensive treatment, the improvement of VAS score and endoscopic Lund-Kennedy score of postoperative compared with preoperative and the same period in the control group were significantly (P < 0.05). The difference of the postoperative asthma control rate of trial group after 1 year and after 3 years, respectively, compared with the same period control group were statistically significant (P < 0.05). The preoperative FEV1, FVC, FEV1/FVC and PEF of trial group compared with preoperative were significantly (P < 0.05). Even the difference of them compared with the same period control group were significantly (P < 0.05), except the FVC in the follow-up 3 years (P = 0.088).@*CONCLUSION@#The CRS may aggravate asthma symptoms and affect negatively the pulmonary function, and poor asthma control or aggravate may exacerbate the CRS in the course of CRS with asthma patient. With ESS based on combined therapy, it can improve the condition of CRS significantly and improve the control of asthma symptoms and pulmonary function else.

Activity and quality comparison of the engineered protein Staphylococcus aureus alpha-hemolysin purified with gel filtration chromatography and Ni-NTA / 生物工程学报

The alpha-hemolysin protein of Staphylococcus aureus, which was expressed in Escherichia coli BL21 (DE3) with recombinant pET32a(+)-alpha-HL plasmid, was purified with gel filtration chromatography (GFC) and Ni-NTA spin columns. The quality and biological characteristic were compared. First, the purified products were analyzed with SDS-PAGE, and the expected protein band was with a molecular mass of 53 kD. Second, protein concentration was determined by the method of Bradford, and the median hemolytic dose potency (HD50) was finally analyzed with rabbit erythrocyte. The protein purified with GFC was 0.337 mg/mL, its hemolysis activity was 1519 HU/mg, and hemolysin yield was 14.04%. Meanwhile, the protein purified with the Ni-NTA Spin Columns was 0.35 mg/mL, its hemolysis activity was 1463 HU/mg, and hemolysin yield was 17.5%, respectively. The results showed that there is no significant difference in the quality, hemolysis activity and yield of the recombinant proteins purified with Ni-NTA spin columns and GFC.

Characterization and Investigation of Staphylococcus aureus zfb Isolates from Bovine Mastitis / 微生物学通报

A Staphylococcus aureus strain, designated zfb, was isolated from a clinical bovine mastitis case of a dairy cow. Staphylococcus aureus zfb can have resistance to methicillin and no lipase contrast by ATCC 25923. The production of the capsule was assessed by the diffuse colonial morphology in serumsoft agar. A mouse infection model was used to determine the LD50 and the invasiveness of SA zfb. The LD50 of SA 25923 to experimental mice was 10-2.5/mL, and the LD50 of SA zfb to experimental mice was 10-4.33/mL. The purpose to detect characteristics of SA zfb makes it an interesting candidate for the preparation and assay of an avirulent mutants against staphylococcal infections and further investigate on pathogenic mechanism.

Fas/APO-1, Caspase-3 and apoptosis of human middle ear cholesteatoma / 中国耳鼻咽喉头颈外科

OBJECTIVE To study the expressions and significance of Fas/APO-1 protein and caspase-3 in human middle ear cholesteatoma and to investigate the relationship between their expressions and the apoptosis of cholesteatoma. METHODS The specimens from the middle ear cholesteatoma tissue of 20 cases and external ear skin of 10 cases were examined by immunohistochemical SP method and TUNEL method. RESULTS There was a significant difference in the expressions of Fas/APO-1 and caspase-3 positive cells between choleseatoma epithelium and normal external ear canal skin(P

Evaluation of the Immunity Effectiveness on the Engineered Subunit Vaccine of Staphylococcus aureus ?-Hemolysin / 中国生物工程杂志

Objective:? -Hemolysin of Staphylococcus aureus,which was expressed in E.coli BL21(DE3) with recombinant pET32a+-?-HL plasmid,was purified with gel filtration chromatography(GFC),and then the engineered subunit vaccine was developed. The immunity effectiveness of this vaccine was evaluated on mouse models.Method:The purified fusion protein was analyzed in SDS-PAGE,and subjected to the evaluation of its median hemolytic dose potency(HD50) was finally analyzed with rabbit erythrocyte. Protein concentration was determined by the method of Bradford. Antibody titers were evaluated on ELISA,and then challenged to gain the immunity protect index.Results:There is an expected protein band with molecular mass of 53kDa in SDS-PAGE,and the concentration is 0.1278mg/ml. The hemolysis activity is 8012.5 HU/mg. There are specific antibodies acquired in blood-serum from mice after vaccined and the antibody titers rising until it has arrive the max,then following down.Conclusion:The purified fusion protein has good fineness and hemolysis activity,the antibody titers initiated by the protein vaccine go with regulation and the immunoprotection is satisfied.

EFFECT OF SCHISTOSOMA JAPONICUM INFECTION ON SERUM TESTOSTERONE LEVELS IN MICE / 中国寄生虫学与寄生虫病杂志

[Objective] To determine the effect of Schistosoma japonicum infection on the testoterone level in the sera from male C57BL/6 mice. [Methods] Radioimmunoassay was used to examine testosterone levels in sera of 9 male mice experimentally infected with Schistosoma japonicum. [Results] The serum testosterone levels reduced significantly in all experimentally infected animals 45 days after infection, as compared with the uninfected controls. [Conclusion] Infection with Schistosoma japonicum decreases testosterone levels in the mouse host.

Epidemiological study of schistosomiasis japonica on liver and spleen with ultrasonographical assessment among inhabitants in Dongting Lake endemic areas / 中国血吸虫病防治杂志

Objective To understand the morbidity of schistosomiasis japonica and health status of the liver and spleen of residents in a village and to evaluate the application of ultrasound on schistosomiasis epidemiology. Methods A total of 454 residents aged 5-65 years were examined by methods of Kato-Katz and ultrasound as well as disease history inquiry. Results The positive rate was 9.38% by stool examination, with no significant difference between males and females. The intensity of the infection among population was 5.70 eggs per gram of stool (EPG), with significant difference between males and females. The stool positive rate and EPG in farmers and students were the highest. Among 10.66% of the males and 8.10% of the females, as well as in 23.81% of the stool positive and 8.31% of the stool negative, the parenchyma of the liver was abnormal(≥GradeⅡ),with significant difference between males and females and between the stool positive and the stool negative. The abnormal rate of liver parenchyma went up with the age. Conclusion Ultrasound can evaluate the health status of inhabitants with schistosomiasis japonica and improve the compliance rate of residents to praziquantel chemotherapy.

EXPERIMENTAL STUDY ON SUSCEPTIBILITY OF PRAZIQUANTEL AGAINST SCHISTOSOMA JAPONICUM IN REPEATED CHEMOTHERAPY AREAS IN DONGTING LAKE REGION / 中国血吸虫病防治杂志

Objective To explore susceptibility of praziquantel(PQT) against Schistosoma japonicum in the repeated chemotherapy areas in Dongting Lake region of China. Methods Sixty mice were divided into two groups, and infected respectively by cercariae released from the infected snails which were collected from new and old endemic areas. After 5 weeks, the mice in each group were divided into control groups and treatment groups (PQT group). The mice in each PQT group were treated with a single oral dose of praziquantel (600 mg/kg). Three weeks post treatment, mice were dissected, and the number of adults, the stool eggs per gram (EPG), the liver EPG and the hatching rates were observed. Results The worm reduction rates of the PQT groups of new and old epidemic areas were 98.24% and 98.71% respectively, and the stool egg reduction rates 99.94% and 99.64%, the liver egg reduction rates 75.85% and 73.10%,and there were no significant differences between the new and old endemic areas. The stool hatching test was positive in the control groups, and negative in the PQT groups. Conclusion Susceptibility of praziquantel against Schistosoma japonicum does not decrease in repeated chemotherapy areas in Dongting Lake region.

Endemic diversity and interventions on schistosomiasis after reserving plain for flooding in Jicheng and Qingshanhu / 中国血吸虫病防治杂志

Objective To understand endemic diversity on schistosomiasis transmission after reserving plain for flooding. Methods In two study pilots, Jicheng and Qingshanhu, epidemiological factors were investigated longitudinally, and the effectiveness of the interventions was evaluated. Results The infection rates of mobile people who engaged in activities in the discarded plain were increased year by year. The density of infected snails was high. The snail habitats increased significantly in Jicheng, but decreased in Qingshanhu. The infection rate and number of livestock pastured in the discarded plain increased. In the discarded plain, most of the mobile people came from the local areas, and main activities for water contact were fishing and pasturing. About 90% of local residents migrated into endemic areas, and the others into non-endemic areas. Conclusions The discarded plains were evolving to a serious transmission zone of schistosomiasis. Interventions combined with agriculture and fisher productions can decrease snail-spreading. Schistosomiasis examination and chemotherapy for the migrants to non-endemic areas are vital.