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Background: Identifying effective biological control agents against fungal pathogens and determining their mechanisms of action are important in the control of plant diseases., Results: In this study, we isolated an endophytic bacterial strain, GsB01, from the branches of asymptomatic Gleditsia sinensis. Multi-locus sequence analysis identified the strain as Bacillus velezensis. GsB01 exhibited significant antifungal activity against Thyronectria austroamericana, the causative agent of G. sinensis wilt. Liquid chromatography-mass spectrometry identified four consistently present antimicrobial compounds in GsB01 metabolite fractions with high antifungal activity: macrolactin A, bacillaene A, surfactin, and iturin. GsB01's active metabolite fractions altered the metabolic profiles of T. austroamericana, disrupting seven pathways, including arginine biosynthesis, nucleotide metabolism, purine metabolism, and the pentose phosphate pathway. Furthermore, absolute quantitative polymerase chain reaction analysis suggested that GsB01 may increase the abundance of endophytic bacteria in G. sinensis. The 16S rRNA amplicon sequencing revealed changes in the endophytic landscape in stems and roots following GsB01 introduction, particularly with significant variation in the dominant bacterial genera within the stems., Conclusion: The study highlights GsB01's potential against plant wilt and suggests that its antifungal activity is achieved by secreting antifungal metabolites. The study also recorded changes in the symbiotic microbiota within trees that had been infected with a pathogenic fungus and subsequently treated with an endophytic antagonistic bacterial strain. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Assessment tools are necessary for the adequate stratification of patients with AIDS-related pneumocystis pneumonia (PCP). The aim of this study was to evaluate the ability of severity assessment scores and inflammation- and nutrition-based parameters for predicting the 180-day mortality of AIDS-related PCP. This was a retrospective cohort study of patients with AIDS-related PCP admitted at the Beijing Di-Tan Hospital. The CURB-65 score, Pneumonia Severity Index (PSI) score, Acute Physiology And Chronic Health Evaluation II (APACHE II) score, C-reactive protein-to-albumin ratio (CAR), procalcitonin, neutrophil-to-lymphocyte ratio, lymphocyte-to-monocyte ratio, and platelet-to-lymphocyte ratio during the first 24 h of intensive care unit admission were analyzed. The prognostic values of the severity assessment scores and biomarkers for 180-day mortality were evaluated using receiver operating characteristic (ROC) curves and integrated discrimination improvement (IDI) indexes. A total of 123 patients with AIDS-related PCP were included. Fifty-five patients were dead, and 68 were still alive at 180 days after admission. CAR, CURB-65, PSI, and APACHE II were independent predictors of 180-day mortality. The optimal cutoff value of CAR was 2.0 mg/g [area under the ROC curve = 0.844, 95% credential interval (CI) = 0.776-0.913], and CAR >2.0 mg/g increased the prognostic value of all three severity assessment scores, with an IDI index of 5.1% for the CURB-65 score, 8.1% for the PSI score, and 4.1% for the APACHE II score (all p  < .05). Combining CAR >2.0 mg/g enhanced the capability of CURB-65, APACHE II, and PSI in predicting the 180-day mortality of patients with AIDS-related PCP.

Objective: To study the clinical value of lymphocyte subsets, immunoglobulins, and complement C3 and C4 in the evaluation of immune status in children with hand-foot-mouth disease (HFMD)., Methods: A total of 282 children with HFMD were enrolled as the HFMD group, and 130 healthy children were enrolled as the healthy control group. The percentages of peripheral CD3 + , CD4 + , and CD8 + T lymphocytes, CD19 + B lymphocytes, and CD56 + natural killer cells were measured. The CD4 + /CD8 + ratio was calculated. The levels of immunoglobulin A (IgA), immunoglobulin M (IgM), immunoglobulin G (IgG), and complement C3 and C4 were measured., Results: The multivariate analysis showed that compared with the healthy control group, the HFMD group had significantly lower percentages of CD3 + , CD4 + , and CD8 + T lymphocytes and levels of complement C3 and C4 (P<0.05), as well as significantly higher percentage of CD56 + natural killer cells and level of IgG (P<0.05). The individual effect analysis showed that the children aged 0-3 years in the HFMD group had a significantly higher CD4 + /CD8 + ratio than the healthy control group (P<0.05); boys aged 0-3 and ≥3 years in the HFMD group had a significantly higher level of IgM than the healthy control group (P<0.05); boys aged ≥3 years and girls aged 0-3 years in the HFMD group had a significantly lower level of IgA than the healthy control group (P<0.05)., Conclusions: Cellular and humoral immunity disorders are observed in children with HFMD. The monitoring of lymphocyte subsets and immunoglobulin levels can provide a laboratory basis for immune status assessment in children with HFMD.

The level of serum lipids is associated with bone mineral density (BMD), an important skeletal trait. Yet the causality has not been determined. Here we performed a Mendelian randomization (MR) analysis to test potential causal links between BMD and lipid profile, i.e., low-density lipoprotein cholesterol (LDC-c), total cholesterol (TC), triglyceride (TG) and high-density lipoprotein cholesterol (HDL-c). We observed causal effect of LDL-c, TC and TG to BMD, and reversely the effect of BMD to HDL-c. We further explored the effect of body mass index (BMI) in these causalities and found that the effect of LDL-c, TC and TG to BMD is independent of BMI. Our findings provided useful information in the clinical relevance of blood lipids on BMD variation and osteoporosis risk., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Objective: To investigate the use of antibiotics in children with community-acquired pneumonia (CAP) in multiple regions of China, and to provide a reference for CAP standard treatment and rational antibiotic use in children., Methods: The medical data of 1 383 children with CAP who were hospitalized in the department of pediatrics in 10 grade A tertiary hospitals from 9 cities between April 14, 2014 and January 1, 2016 were reviewed, to analyze the status of antibiotic use in hospitalized children in North China, Northeast China, East China, and South China., Results: The overall rate of antibiotic use in children with CAP was 89.08%, with 88.7% in North China, 95.5% in Northeast China, 83.3% in East China, and 86.6% in South China. The main types of antibiotics used were cephalosporins, macrolides, compound preparations of β-lactam antibiotics, polyphosphoric broad-spectrum antibiotics and other β-lactam antibiotics. The selection of antibiotics was generally rational, but antibiotics were still used in some patients with viral infection alone or a combined use of ≥2 kinds of antibiotics were noted in some patients with infection caused by one kind of pathogen. Irrational antibiotic use was observed in 131 children (10.63%)., Conclusions: There are high rates of antibiotic use and irrational use of antibiotics among children with CAP. Standard management of antibiotic use in children with CAP should be strengthened.

Objective: Observation of the effect of uvuplopalatal flap for OSAHS patients with anterior and posterior pharyngeal stenosis. Method: Fifty-six patients with OSAHS who were mainly anterior posterior stenosis in the velopharyngeal plane were selected. Patients with suspending uvuplopalatal flap were used as observation group (30 cases). Patients with UPPP were used as control group (26 cases). The investigation and comparison of AHI, SaO₂, scars, throat foreign body sensation, pharynx desiccation, pharynx' s tightness, pharynx discomfort and surgical satisfaction were investigated at 6-8 months after the operation. Restul: At 6-8 months after operation, the AHI and SaO₂ of the two groups were significantly improved compared with those before operation( P <0.01). However, there were significant differences( P <0.01) among the indexes of scar formation, throat foreign body, sensation pharynx desiccation, pharynx' s tightness, pharynx discomfort and surgical satisfaction. Conclusion: In the OSAHS patients with anterior and posterior pharyngeal stenosis, the effect of UPF was better than that of UPPP among the indexes of scar formation, pharyngeal symptoms and surgical satisfaction., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

Background: The expression of 4-1BB on peripheral regulatory T cells (Tregs) and conventional T cells (Tconvs) in coronary artery disease (CAD) patients is unknown. We aimed to investigate the expression and clinical correlations of 4-1BB on peripheral Tregs and Tconvs in CAD patients., Methods: Flow cytometry analysis was used to analyze 4-1BB expression on peripheral Tregs and Tconvs. We compared the percentages of 4-1BB on Tregs and Tconvs in the control (ctrl) group, the stable ischemic heart disease (SIHD) group, and the acute coronary syndrome (ACS) group. The correlations of 4-1BB expression on Tregs and Tconvs with the Gensini score and CRP were examined in the ACS group. The value of 4-1BB percentage on Tregs for predicting CAD in this cardiovascular risk population was also analyzed., Results: A total of 71 participants were enrolled in this study. In all the groups, the percentages of 4-1BB on Tregs were significantly higher than on Tconvs (all P < .05). After adjusting for sex, age, SBP, HbA1c and LDL, 4-1BB percentages on Tregs and Tconvs were significantly higher in the SIHD and ACS groups compared with the ctrl group (all P < .05). The ratio of 4-1BB percentage on Tregs to 4-1BB percentage on Tconvs was higher in the ACS group compared with the ctrl group (P = .010). In the ACS group, CRP was negatively correlated with the Tregs percentage (in CD4+ T cells) and the Tregs percentage to Tconvs percentage ratio. The Gensini score was positively correlated with the 4-1BB percentage on Tregs in the ACS group. Linear regression analysis showed 4-1BB percentage on Tregs independently predicted the Gensini score. Binary logistic regression showed CRP, HbA1c and 4-1BB percentage on Tregs independently predicted the development of CAD (SIHD+ACS) in the whole population., Conclusion: 4-1BB expression on peripheral Tregs and Tconvs was increased in SIHD and ACS patients. 4-1BB percentage on Tregs positively correlated with the severity of coronary artery stenosis in ACS patients. 4-1BB percentage on Tregs independently predicted the severity of coronary artery stenosis in an ACS population and development of CAD in a cardiovascular risk population., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Insulin-like growth factor 1 (IGF1) is a key factor for tissue growth and fuel metabolism. The potential function of central IGF1 remains unclear. We previously observed that IGF1 expression is increased in the hypothalamus of obese mice lacking STAT5 in the central nervous system (CNS). In this study, we explored the potential metabolic function of central IGF1 by intracerebroventricular (ICV) injection of IGF1, over-expression of central IGF1 by administering an adeno-associated virus (AAV), and ICV injection of an anti-IGF1 antibody. Mice that over-expressed central IGF1 displayed increased appetite, improved glucose tolerance and insulin sensitivity, decreased Pomc levels in the hypothalamus, and increased UCP1 expression in brown fat tissue. This is the first study demonstrating that central IGF1 regulates several important metabolic functions.

Objective: To study the spatial and temporal mode of infectious TB transmission in Guangxi Zhuang Autonomous Region (Guangxi). Methods: Data related to infectious TB case (Include smear and/or culture positive patients) in Guangxi were collected from the National Notifiable Disease Reported System (NNDRS) from 2010 to 2015. Spatial-temporal analysis and prediction were performed by SaTScan 7.0.2, GeoDa 1.8.12, R program v 3.3.1 and SPSS 19.0 software, using the time series model, Moran's I global and local spatial autocorrelation (Empirical Bayes adjustment). Kulldorff 's space-time scan statistics displayed by R software was used to identify the temporal and spatial trend of TB. Results: The total number of infectious TB cases, collected from NNDRS was 76 151, and showing a decreasing trend on annual incidence (value of Chi-square for Linear trend=3 464.53, P -value=0.000). The forecast value of TB cases in 2016 was 7 764 (4 971-10 557), with peak in March, analyzed through the Winters'multiplicative model. The Moran's I global Statistics was greater than 0 (0.257-0.390). TB cluster seemed to have been existed for several years. The most significant hot spots seemed to be mainly located in the central and western parts of Guangxi, shown by local spatial autocorrelation statistics and the result from space-time scanning.Counties or districts that located in the east parts of Guangxi presented the low-low relation (significant cold spots). The situation of infectious TB seemed migratory. Conclusions: Our data showed an annual decreasing trend of incidence on infectious TB with temporal concentration in spring and summer. Main clusters (hot spots) were found to be located in the central and western parts of Guangxi. Hopefully, our findings can provide clues to uncover the real mode of TB transmission at the molecular-biological level.

In the last decade, numerous microRNAs (miRNAs) have been identified in diverse virus families, particularly in herpesviruses. Gallid alphaherpesvirus 2 (GaHV2) is a representative oncogenic alphaherpesvirus that induces rapid-onset T-cell lymphomas in its natural hosts, namely Marek's disease (MD). In the GaHV2 genome there are 26 mature miRNAs derived from 14 precursors assembled into three clusters, namely the Meq-cluster, Mid-cluster and LAT-cluster. Several GaHV2 miRNAs, especially those in the Meq-cluster (e.g. miR-M4-5p), have been demonstrated to be critical in MD pathogenesis and/or tumorigenesis. Interestingly the downstream Mid-cluster is regulated and transcribed by the same promoter as the Meq-cluster in the latent phase of the infection, but the role of these Mid-clustered miRNAs in GaHV2 biology remains unclear. We have generated the deletion mutants of the Mid-cluster and of its associated individual miRNAs in GX0101 virus, a very virulent GaHV2 strain, and demonstrated that the Mid-clustered miRNAs are not essential for virus replication. Using GaHV2-infected chickens as an animal model, we found that, compared with parental GX0101 virus, the individual deletion of miR-M31 decreased the mortality and gross tumour incidence of infected chickens while the deletion individually of miR-M1 or miR-M11 unexpectedly increased viral pathogenicity or oncogenicity, similarly to the deletion of the entire Mid-cluster region. More importantly, our data further confirm that miR-M11-5p, the miR-M11-derived mature miRNA, targets the viral oncogene meq and suppresses its expression in GaHV2 infection. We report here that members of the Mid-clustered miRNAs, miR-M31-3p and miR-M11-5p, potentially act either as oncogene or tumour suppressor in MD lymphomagenesis.

In this paper, the laboratory-scale permeable pavement layers, including a surface permeable brick layer, coarse sand bedding layers (thicknesses = 2, 3.5, and 5 cm), and single-graded gravel sub-base layers (thicknesses = 15, 20, 25, and 30 cm), were built to evaluate stormwater infiltration and surface runoff pollution reduction performance. And, the infiltration rate (I) and concentrations of suspended solids (SS), total phosphorus (TP), chemical oxygen demand (COD), ammonia nitrogen, and total nitrogen (TN) were measured under the simulated rainfall intensity of 72.4 mm/h over duration of 60 min. The results indicate that the thickness factor primarily influences the infiltration rate and pollutant removal rate. The highest steady infiltration rate was for surface brick layer 51.0 mm/h, for 5-cm sand bedding layer 32.3 mm/h, and for 30-cm gravel sub-base layer 42.3 mm/h, respectively. The SS average removal rate was relative higher (79.8 ∼ 98.6 %) for all layers due to the interception and filtration. The average removal rates of TP and COD were for surface layer 71.2 and 24.1 %, for 5-cm bedding layer 54.8 and 9.0 %, and for 20-cm sub-base layer 72.2 and 26.1 %. Ammonia nitrogen and TN cannot steadily be removed by layers according to the experiment results. The optimal thickness of bedding sands was 5 cm, and that of sub-base gravels was 20 ∼ 30 cm.

Marek's disease virus (MDV) is an important oncogenic alphaherpesvirus that induces rapid-onset T-cell lymphomas in its natural hosts. The Meq-clustered miRNAs encoded by MDV have been suggested to play potentially critical roles in the induction of lymphomas. Using the technique of bacterial artificial chromosome mutagenesis, we have presently constructed a series of specific miRNA-deleted mutants and demonstrate that these miRNAs are not essential for replication of MDV and have no effects on the early cytolytic or latent phases of the developing disease. However, compared to the parental GX0101, mortality of birds infected with the mutants GXΔmiR-M2, GXΔmiR-M3, GXΔmiR-M5, GXΔmiR-M9 and GXΔmiR-M12 was reduced from 100 % to 18 %, 30 %, 48 %, 24 % and 14 %, coupled with gross tumour incidence reduction from 28 % to 8 %, 4 %, 12 %, 8 % and 0 %, respectively. Our data confirm that except for mdv1-miR-M4, the other Meq-clustered miRNAs also play critical roles in MDV oncogenesis. Further work will be needed to elucidate the miRNA-mediated regulatory mechanisms that trigger the development of MD lymphomas., (© 2015 The Authors.)

Each Caulobacter cell cycle involves differentiation and an asymmetric cell division driven by a cyclical regulatory circuit comprised of four transcription factors (TFs) and a DNA methyltransferase. Using a modified global 5' RACE protocol, we globally mapped transcription start sites (TSSs) at base-pair resolution, measured their transcription levels at multiple times in the cell cycle, and identified their transcription factor binding sites. Out of 2726 TSSs, 586 were shown to be cell cycle-regulated and we identified 529 binding sites for the cell cycle master regulators. Twenty-three percent of the cell cycle-regulated promoters were found to be under the combinatorial control of two or more of the global regulators. Previously unknown features of the core cell cycle circuit were identified, including 107 antisense TSSs which exhibit cell cycle-control, and 241 genes with multiple TSSs whose transcription levels often exhibited different cell cycle timing. Cumulatively, this study uncovered novel new layers of transcriptional regulation mediating the bacterial cell cycle.

Background: Treatment of uremia is now dominated by dialysis; in some cases, patients are treated with dialysis for decades, but overall outcomes are disappointing. A number of studies have confirmed the relevance of several experimental insights to the pathogenesis of uremia, but the specific biomarkers of uremia have not been fully elucidated. To date, our knowledge about the alterations in DNA 5-hydroxymethylcytosine (5-hmC) in uremia is unclear, to investigate the role of DNA 5-hmC in the onset of uremia, we performed hMeDIP-chip between the uremia patients and the normal controls from the experiment to identify differentially expressed 5-hmC in uremia-associated samples., Methods: Extract genomic DNA, using hMeDIP-chip technology of Active Motif companies for the analysis of genome-wide DNA 5-hmC, and quantitative real-time PCR confirmation to identify differentially expressed 5-hmC level in uremia-associated samples., Results: There were 1875 genes in gene Promoter, which displayed significant 5-hmC differences in uremia patients compared with normal controls. Among these genes, 960 genes displayed increased 5-hmC and 915 genes decreased 5-hmC. 4063 genes in CpG Islands displayed significant 5-hmC differences in uremia patients compared with normal controls. Among these genes, 1780 genes displayed increased 5-hmC and 2283 genes decreased 5-hmC. Three positive genes, HMGCR, THBD, and STAT3 were confirmed by quantitative real-time PCR., Conclusion: Our studies indicate the significant alterations of 5-hmC. There is a correlation of gene modification 5-hmC in uremia patients. Such novel findings show the significance of 5-hmC as a potential biomarker or promising target for epigenetic-based uremia therapies.

Objective: This study aimed to evaluate the diagnostic value of a 24-hour esophageal combined multichannel intraluminal impedance pH (24 h our MII-pH) monitoring in children with gastroesophageal reflux (GER) disease and recurrent pneumonia., Methods: A total of 17 cases with a suspected diagnosis of GER disease children with unexplained recurrent pneumonia underwent a 24-hour MII-pH monitoring to analysis of the nature and characteristics of reflux., Results: In the 17 cases of GER disease with recurrent pneumonia, 11 (65%) were confirmed positively by a 24-hours of MII-pH monitoring. A total of 853 reflux cycle were detected, of which 65.3% were acid refluxes predominantly occurring within 2 hours after meal, and 71.6% were mixed refluxes. Refluxes occurred most frequently in the supine position, followed by the lateral position and the prone position. Distal reflux was the predominant form of reflux. According to the Biox-Ochoa classification, 73% of cases (8/11) were severe GER., Conclusions: MII-pH monitoring may effectively characterize refluxate properties and thereby increase the detection rate of GER disease.

The microRNA (miRNA) mdv1-miR-M4, a functional miR-155 ortholog encoded by oncogenic Marek's disease virus (MDV), has previously been suggested to be involved in MDV pathogenesis. Using the technique of bacterial artificial chromosome mutagenesis, we have presently evaluated the potential role of mdv1-miR-M4 in the oncogenesis of the very virulent (vv) MDV strain GX0101. Unexpectedly, deletions of the Meq-cluster or mdv1-miR-M4 alone from the viral genome strongly decreased rather than abolished its oncogenicity. Compared to GX0101, mortalities of mutants GXΔmiR-M4 and GXΔMeq-miRs were reduced from 100% to 18% and 4%, coupled with the gross tumor incidence reduction from 28% to 22% and 8%, respectively. Our data suggests that the mdv1-miR-M4 is possibly an important regulator in the development of Marek's disease (MD) lymphomas but is not essential for the oncogenicity of vvMDV. In addition, some of the other Meq-clustered miRNAs may also play potentially critical roles in vvMDV induction of lymphomas., (© 2013 Published by Elsevier Inc.)

Background: Community-acquired pneumonia in children is common in China. To understand current clinical characteristics and practice, we conducted a cross-sectional study to analyze quality of care on childhood pneumonia in eight eastern cities in China., Methods: Consecutive hospital records between January 1, 2010 and December 31, 2010 were collected from 13 traditional Chinese medicine (TCM) and western medicine (WM) hospitals in February, May, August, and November (25 cases per season, 100 cases over the year), respectively. A predesigned case report form was used to extract data from the hospital medical records., Results: A total of 1298 cases were collected and analyzed. Symptoms and signs upon admission at TCM and WM hospitals were cough (99.3% vs. 98.6%), rales (84.8% vs. 75.0%), phlegm (83.3% vs. 49.1%), and fever (74.9% vs. 84.0%) in frequency. Patients admitted to WM hospitals had symptoms and signs for a longer period prior to admission than patients admitted to TCM hospitals. Testing to identify etiologic agents was performed in 1140 cases (88.4%). Intravenous antibiotics were administered in 99.3% (595/598) of cases in TCM hospitals and in 98.6% (699/700) of cases in WM hospitals. Besides, Chinese herbal extract injection was used more frequently in TCM hospitals (491 cases, 82.1%) than in WM hospitals (212 cases, 30.3%) (p < 0.01). At discharge, 818 cases (63.0%) were clinically cured, with a significant difference between the cure rates in TCM (87.6%) and WM hospitals (42.0%) (OR = 9.8, 95% confidence interval (CI): 7.3 ~ 12.9, p < 0.01). Pathogen and previous medical history were more likely associated with the disappearance of rales (OR = 7.2, 95% CI: 4.8 ~ 10.9). Adverse effects were not reported from the medical records., Conclusions: Intravenous use of antibiotics is highly prevalent in children with community-acquired pneumonia regardless of aetiology. There was difference between TCM and WM hospitals with regard to symptom profile and the use of antibiotics. Intravenous use of herbal injection was higher in TCM hospitals than in WM hospitals. Most of the cases were diagnosed based on clinical signs and symptoms without sufficient confirmation of aetiology. Audit of current practice is urgently needed to improve care.

To study the correlation between ELISA and IFA tests in detection of ALV-A/B antibody in chicken sera, ELSA S/P values and IFA titers for different serum samples were measured and statistically analyzed. The results indicated that there was a strong positive correlation between ELISA S/P values and IFA titers (r = 0.97435, P < 0.001). Because the positive correlation between ELISA and IFA was so strong and antibody positive rates were identical in two tests, it suggested that IFA could be used as a alternative method to replace ELISA kit when only limited numbers of samples to be tested to reduce the cost and increase the sensitivity.

Immunosenescence is a result of progressive decline in immune system function with advancing age. Epidermal Langerhans cells (LCs), belonging to the dendritic cell (DC) family, act as sentinels to play key roles in the skin immune responses. However, it has not been fully elucidated how aging affects development and function of LCs. Here, we systemically analyzed LC development and function during the aging process in C57BL/6J mice, and performed global microRNA (miRNA) gene expression profiles in aged and young LCs. We found that the frequency and maturation of epidermal LCs were significantly reduced in aged mice starting at 12 months of age, while the Langerin expression and ability to phagocytose Dextran in aged LCs were increased compared to LCs from < 6 month old mice. The migration of LCs to draining lymph nodes was comparable between aged and young mice. Functionally, aged LCs were impaired in their capacity to induce OVA-specific CD4+ and CD8+ T cell proliferation. Furthermore, the expression of miRNAs in aged epidermal LCs showed a distinct profile compared to young LCs. Most interestingly, aging-regulated miRNAs potentially target TGF-β-dependent and non- TGF-β-dependent signal pathways related to LCs. Overall, our data suggests that aging affects LCs development and function, and that age-regulated miRNAs may contribute to the LC developmental and functional changes in aging.

In order to clarify Avian leukosis virus (ALV) characteristics from Chinese native chicken breeds, three ALV JS11C1, JS11C2 and JS11C3 were isolated from Chinese native breed "luhua" by inoculation of DF1 cell culture and detection of p27 antigen. Using PCR amplification of env gene, the amplified gp85 genes were analyzed and compared to all six chicken ALV subgroups reported. The gp85 genes of these three viruses were 1 005bp in length and encoded 335 amino acids, and the gp37 genes were 609bp and encoded 203 amino acids. The homology of gp85 among these three isolated strains was 91.9%-97.0%. Comparing to 18 stains of subgroup A, B, C, D, E published in GenBank, the homology was only in the range of 77.7%-84.6%, significantly lower than the gp85 homology observed within the common chicken subgroups A (88.2%-98.5%), B (91.6%-98.8%), and E (97.9%-99.4%). The gp85 homology compared with subgroup J was only 34.2%-36.5%. These results suggested that three isolated strains from Chinese native breed "luhua" belong to a new subgroup different from all six known subgroups from Chickens, and thus designated as subgroup K.

In chicken fibroblast cell (CEF) cultures with antiserum against Newcastle disease virus (NDV) strain TZ060107, the virus was passed serially for 50 passages in 3 independent lineages. HN and F genes were amplified and sequenced every 10 passages. The derived virus A1-50 with most mutations among 3 lineages was further passed for another 50 passages in CEF with or without antiserum against A1-50, each in 3 independent lineages. Sequence comparisons for HN and F genes of 60, 70, 80, 90 and 100 passages indicated that the ratio of nonsynonymous mutations (NS) vs synonymous mutations (S) for HN genes in the lineages passed with antiserum against A1-50 was 5.25, which was obviously higher than 2. 375 of NS/ S in the lineages without the antiserum. The stable NS mutations occurred in the first 50 passages with the antiserum against the original TZ060107 were still maintained and one more new stable NS mutation appeared. For the F gene, 3 new stable NS mutations occurred during the second 50 passages in lineages with antiserum against A1-50 when the original NS mutations obtained in the first 50 passages with antiserum against TZ060107 still existed. Cross hemagglutination inhibition (HI) between original virus and its derivative viruses indicated that the more continuous passages in cell culture with antiserum passed, the bigger difference of antigenicity between the virus and the original virus had.

LG1 strain of avian influenza virus H9N2 was passaged continuously for 40 generations in chicken embryos with anti-LG1 maternal antibodies in 4 parallel experiments, of which 3 experiments had a stable mutation of "G" to "A" at #99 of the neuraminidase gene(NA)from the 20th passage resulting in a change of Met to Ile and 2 had a stable mutation of "A" to "G" at #473 of the NA gene from the 30th passage resulting in a change of Asn to Ser which occurred in the 50th passage of another experiment. Eighty continuous passages in chicken embryos without antibody did not have the same mutation, indicating that the mutations of the 2 positions were associated with selective pressure of antibodies. Analysis of the ratios of nonsynonium (NS) vs synonium (S) mutations of nucleic acids demonstrated that NS/S of 4 parallel experiments with antibodies was 4.6 (32/7) compared with 2.0 (16/8) of the 2 experiments without antibodies and this significant difference implied the selective pressure of antibodies.

To study the correlation between 50% tissue-culture infective dose (TCID50) value and p27 antigen S/P value of Avian leukosis virus subgroup J and discuss their significance, chicken embryo fibroblast (CEF) cells were inoculated with Avian leukosis virus subgroup J strain NX0101 and samples were tested continuously for ten days after changing maintenance media. The correlation between TCID50 and p27 antigen S/P value of ten days were then analysized. Simultaneously, DF-1 cells were inoculated with NX0101 and passaged to 20 generations. Samples taken from 1st generation, 5th generation, 10th generation, 15th generation and 20th generation were tested for the TCID50 titer and the p27 antigen S/P value separately. A significant Pearson correlation was found between them in CEF cells (r = 0.85277; P < 0.0001) and in DF-1 cells (r = 0.93000; P = 0.0220). This study provided an important parameter for predicting TCID50 by detecting the p27 antigen S/P value.

The purpose of this study was to compare the whole genome sequences and replication dynamics in cell cultures of two Avian leukosis viruses of subgroup B (ALV) isolates, SDAU09E3 and SDAU09C2. Comparison of the amino acid sequences indicated that the gp85 identity of these two subgroup B isolates was 95.4%, the identity with other three ALV-B reference strains was 91.0%-94.9%, and less than 87.9% with ALV subgroup A, C, D, E and J. Comparison of the nucleotide sequence of gag and pol genes indicated that homologies of gag gene and pol gene of these two ALV-B isolates with all compared reference strains of different subgroups were above 93%. Homologies of LTR sequence of these two ALV-B isolates with other exogenous ALVs subgroups A, B, C, D and J were 72.6%-88.3%, but only 51.5% when compared with endogenous ALV subgroup E. The identity of LTR between these two ALV-B strains was only 74.8%, which was far lower than the identity of other genes. The identity of U3 region of LTR between these two ALV-B isolates was only 68.8% and there were obvious differences in the number CAAT Boxes. Replication dynamics in DF-1 cell indicated that the value of TCID50 was similar between 2 isolates but the concentration of nucleocapsid protein p27 antigen of SDAU09E3 was significantly higher than SDAU09C2 in cell culture supernatant, which indicated there was no parallel relationship between p27 antigen concentration and infectious virus particles. Whether such difference was resulted from the diversity of U3 region of LTR, further studies with their recombinant infectious clones is necessary.

Marek's disease virus (MDV) is a long-recognized oncogenic herpesvirus, which induces lymphoma in its natural host that can be prevented by vaccination. MDV infection provides an excellent biological model for investigating the biology, genetics and immunology of viral oncogenesis. Recently discovered microRNAs (miRNAs) in the MDV genome have been suggested to have regulatory roles during MDV oncogenesis. We have examined the expression profiles of all 22 previously reported miRNAs encoded by MDV-1 in chickens artificially challenged with MDV-GX0101. We found that a subset of the miRNAs was differentially expressed during different phases of the developing disease. These miRNAs show early or late expression during disease progression, accompanied by obvious tissue-specific and differential expression patterns. This temporal and differential tissue distribution suggest that these miRNAs may perform different regulatory roles in switching from latency to lytic replication, immunosuppression, neoplastic transformation or other aspects of lymphoma formation. These reported in vivo expression profiles indicate the potentially functional MDV-1-encoded miRNAs that should be selected for further investigation of their functions in MDV oncogenesis.

The predominant field strains of Marek's disease virus in Guangxi were clearly different from the vaccine strain CVI988/Rispens based on sequencing of the envelope glycoprotein I (gI), glycoprotein E (gE) and oncogenic meq genes. These differences may be partly responsible for the most recent outbreaks in Guangxi.

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.

MDV-GX0101 is a field strain of Marek's disease virus with a naturally occurring insertion of the reticuloendotheliosis virus (REV) LTR fragment. In order to study the biological properties of REV-LTR insertion in the MDV genome, we constructed a full-length infectious BAC clone of MDV-GX0101 strain and deleted the LTR sequences by BAC mutagenesis. The pathogenic properties of the LTR-deleted virus were evaluated in infected SPF birds. The study demonstrated that the LTR-deleted virus had a stronger inhibitory effect on the growth rates of the infected birds and induced stronger immunosuppressive effects. Surprisingly, however, the ability for horizontal transmission of the LTR-deleted virus appeared to be significantly weaker than its parental LTR-intact virus. Even though the precise molecular mechanisms are still not clear, the results of our studies demonstrate that the retention of the REV-LTR in the MDV genome decreases its pathogenic effects but increases its potential for horizontal transmission., (Copyright 2009 Elsevier Inc. All rights reserved.)

Ethnopharmacological Relevance: Radix astragali, Radix codonopis, Herba epimedii and Radix glycyrrizae are 4 plants commonly used in Chinese traditional medicine or veterinary medicine to improve immune functions against chronic diseases in humans and animals., Aim of the Study: We compared immunological enhancement by 4 herbal extracts in clinical healthy chickens or immunosuppressed chickens singly and in combination., Materials and Methods: Water extracts of 4 herbs individually and in different combinations were supplemented in drinking water. Hemagglutination inhibition (HI) antibody titers against Newcastle disease virus (NDV) and H5 avian influenza virus (H5-AIV) after vaccination were measured as indicators to evaluate immunological stimulation across groups supplemented with different herbal extracts. The experiments were conducted in both clinically healthy chickens and chickens with immunosuppression induced by reticuloendotheliosis virus (REV) infection., Results: In clinically healthy chickens HI antibody titers against NDV and H5-AIV after vaccination were not influenced by supplementation with the herbal extracts of Radix astragali, Radix codonopis, Herba epimedii and Radix glycyrrizae in drinking water. In chicks with REV-induced immunosuppression, however, supplementation of some herbal extracts significantly increased HI antibody titers to NDV and H5-AIV when compared to the immunosuppressed control group (P<0.01), but the titers were still lower than those in chicks not infected by REV. The 4 herbal mixtures produced the best enhancement among various combinations. The components of the herbal extract were water soluble and treatment by ether had no influence on immunological enhancement. The molecular weights of the active components of the herbal extracts were in the range of 10,000-100,000 Da., Conclusion: Our results show that the herbal extract supplementation in drinking water can induce an immune stimulation response in immunosuppressed chickens. It suggests that chickens with REV infection-induced immunosuppression could be used as an experiment model for determination of immunological enhancement effects of some herbal components., (Copyright 2009 Elsevier Ireland Ltd. All rights reserved.)

By inoculation of blood samples in DF-1 (C/E) cell culture, an exogenous avian leukosis virus (ALV) strain SDAU09C2 was isolated from a breeder farm of Chinese native breed "Luhua" in Shandong province. Comparisons of the amino acid sequence of env gene gp85 from the isolate with those from other ALV reference strains of different subgroups indicated that SDAU09C2 had the highest gp85 identity to two reference strains of subgroup B of 92.5%. Its gp85 identity to other chicken ALV subgroups A, C, D, E was in the range of 73.2%-87.9%. The identity to subgroup J was only 30.3%-32.4%. This is the first report on isolation and identification of ALV-B and its gp85 from Chinese native breed chickens.

Objective: To investigate the changes of endocrine, cyclic nucleotide and immune systems in subjects of yang deficiency constitution, and to explore the relationship among characteristics and causes of yang deficiency constitution, the physiological and biochemical parameters., Methods: Based on the diagnostic criteria for the clinical epidemiological investigation, sixty subjects of yang deficiency constitution and fifty of normal constitution were selected. Eight milliliters venous blood were taken from overnight fasted subjects at 8:00-9:00. The sera were obtained by centrifugation of the blood at the speed of 3000 r/min for 5 minutes, and they were stored at -70 degrees C until use. The serum levels of corticosterone, cortisol, adrenocorticotrophic hormone (ACTH), cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), free triiodothyronine (FT3), free thyroxine (FT4), thyrotropic-stimulating hormone (TSH), interleukin-1beta and interleukin-2 were measured by enzyme-linked immunosorbent assay; the cAMP/cGMP ratio was also computed; and the differences of the above indexes were compared between the two types of subjects., Results: The serum levels of corticosterone, interleukin-1beta, TSH and cAMP/cGMP ratio of yang deficiency constitution significantly increased as compared with those of normal constitution, and the serum levels of cortisol, ACTH, cGMP and FT4 of yang deficiency constitution significantly decreased in comparison with those of normal constitution., Conclusion: Subjects of yang deficiency constitution may be not only related to hypothalamic-pituitary-adrenal axis and hypothalamic-pituitary-thyroid axis, but also related to the functional disorders of cyclic nucleotide and immune systems.

Objective: To study the distribution and drug resistance of the isolated bacteria from children with acute respiratory infection (ARI) in Dalian., Methods: Between January 2006 and February 2007, 930 children with ARI were enrolled, including 364 with acute upper respiratory infection (AURI), and 566 with acute lower respiratory infection (ALRI). The AURI children, who did not receive antimicrobial agent treatment or received oral antimicrobial agents 1-2 times, had bacterial cultures of pharyngeal swab. The ALRI children, who received intravenous antibacterial agents more than 3 days, had bacterial cultures of sputum and bronchoalveolar lavage fluid (BALF). Isolated bacteria were identified by the ATB system (Bio-Merieux, France). Antimicrobial susceptibility testing was carried out by means of Kirby-bauer., Results: A total of 404 isolates (43.4%) were identified. Haemophilus influenzae, Haemophilus parainfluenzae and Streptococcus pneumoniae accounted for 22.5%, 12.1% and 7.4% respectively. In the isolates from AURI, Haemophilus influenzae, Haemophilus parainfluenzae and Streptococcus pneumoniae accounted for 43.9%, 22.0% and 9.1% respectively; Escherichia coli, Klebsiella pneumonia and Nonfermenters accounted for 4.5%, 8.3% and 3.0% respectively. In the isolates from ALRI, Haemophilus influenzae, Haemophilus parainfluenzae and Streptococcus pneumoniae accounted for 12.1%, 7.4% and 6.6% respectively; Escherichia coli, Klebsiella pneumoniae and Nonfermenters accounted for 16.9%, 13.2% and 21.8% respectively. The resistant rates of Haemophilus to ampicillin and TMP-SMZ were 29.3% and 32.9% respectively, and to amoxicillin-clavulanic acid, cefalotin, cefaclor, cefuroxime and cefotaxime were 12.1%, 10.0%, 10.0%, 11.4% and 5.7%, respectively. The resistant rate of Haemophilus to ampicillin, amoxicillin-clavulanic acid, cefaclor, tetracycine and TMP-SMZ in the ALRI group were significantly higher than that in the AURI group (P<0.05 or 0.01)., Conclusions: In Dalian, Haemophilus was the main isolate of children with ARI. The distribution of bacteria was different between ALRI and AURI. In ALRI, Gram-negative bacilli were in a higher proportion, and the resistant rates of Haemophilus influenzae and Haemophilus parainfluenzae to ampicillin, amoxicillin-clavulanic acid and cefaclor were higher.

Two Avian leukosis viruses of subgroup J (ALV-J) were isolated from layers by inoculating the sample into chicken embryo fibroblast (CEF) cells and by indirect fluorescent assay (IFA) with ALV-J specific monoclonal antibody JE-9. Sequence comparison indicated the gp85 identities were only in the ranges of 83.4%-87.3% compared with five international reference strains and 86.4%-89.6% compared with eight Chinese strains isolated from white meat-type chickens. The gp37 identities were in the range of 91.8-96.4% compared with the five international strains and 93.4-95.9% compared with the eight domestic strains. When compared with the above thirteen strains, two layers isolates were more close to the prototype HPRS-103 and had less deletion in 3'-Ter than those strains. All strains isolated from white meat-type chickens in China had a deletion in the "E" region of 3'-Ter except these two isolates, suggesting these two ALV-J isolates from layers have different evolution origins from other Chinese isolates from white meat-type chickens.

Three cases of Newcastle disease virus (NDV) found in nature had the lentogenic motif (112)G-R-Q-G-R-L(117) in their fusion protein cleavage sites. However, both intracerebral pathogenicity and intravenous pathogenicity indexes showed that these NDV isolates were virulent. In comparison with the LaSota live virus vaccine, these viruses had significant genetic variations in the hemagglutinin-neuraminidase gene.

A field strain C14 of chicken infectious anemia virus (CAV) was isolated from a 14 day-old broiler flock with growth runting syndromes. Antibody reactions to inactivated vaccines to avian influenza viruses (AIV) were suppressed in SPF chickens inoculated with C14 strain CAV at 1 day-old. Also C14 strain CAV and reticuloendotheliosis Virus demonstrated a synergism in immunosuppression when chickens were infected with both virus. The viral genomic DNA was amplified by PCR in 3 overlapped fragments and PCR products were cloned into T-vector plasmid for sequencing. The sequencing results indicated that the total genome of C14 strain CAV was 2298bp, it contained 3 overlapped ORF and 1 non-coding regulation fragment. Its whole genome had 97.2% - 99.2% of homogeneity to other several published CAV reference strains. Sequence data indicated that there are many motifs in the non-coding area of about 400bp as the binding sites for transcriptional factors. All these motifs were very conservative. There were some mutations in 3 genes VP1, VP2 and VP3. Relatively, VP1 was less conservative than VP2 and VP3. Among different strains, mutations in these 3 genes were not correlated.

The influences of avian reovirus (ARV) infection at 1 day of age on bursa development, antibody responses after vaccinations to avian influenza virus (AIV), Newcastle disease virus (NDV) and infectious bursal disease virus (IBDV), and pathogenecity of virulent IBDV (v-IBDV) were studied in chickens of SPF-origin. The results indicated that LY strain ARV infection in 1-day-old chickens caused atrophy of the Bursa and decreased lymphocyte numbers in the bursa, but it gave no significant negative effects on growth rates and antibody titers to AIV and NDV after vaccination. LY strain ARV infection decreased antibody titers to IBDV in B87 vaccinated birds but all vaccinated birds infected with ARV were still full protected from death or clinical syndromes after v-IBDV challenge. Although all B87-vaccinated birds were full protected from death after v-IBDV challenges, the antibody titers to AIV or NDV after vaccinations with inactivated vaccines were significantly lower in v-IBDV challenged birds than controls. Supprisingly, ARV infection at ages of 1-7 days could compromise the immune suppression induced by v-IBDV in B87-vaccinated birds as HI antibody titers to AIV and NDV in ARV-infected groups were significantly higher than chicknes with no ARV infection. A discussion was made on the interactions between ARV infection and vaccine IBDV or v-IBDV to explain such sophisticated phenomena in the bird experiments.

Objective: To investigate the quantities of bone marrow CD5+ B lymphocytes in the patients with autoimmune hemocytopenia and the relationship between quantities of CD5+ B lymphocytes and clinical or laboratorial parameters., Methods: Quantities of CD5+ B lymphocytes in the bone marrow of 14 patients with autoimmune hemolytic anemia (AIHA) or Evans syndrome, 22 immunorelated pancytopenia (IRP) patients, and 10 normal controls were assayed by flow cytometry. The correlation between their clinical or laboratorial parameters and CD5+ B lymphocytes was analyzed., Results: The quantity of CD5+ B lymphocytes of AIHA/Evans syndrome (34.64% +/- 19.81%) or IRP patients (35.81% +/- 16.83%) was significantly higher than that of normal controls (12.00% +/- 1.97%, P < 0.05). However, there was no significant difference between AIHA/Evans syndrome and IRP patients (P > 0.05). In all hemocytopenic patients, the quantity of bone marrow CD5+ B lymphocytes showed significantly negative correlation with serum complement C3 level (r = -0.416, P < 0.05). In the patients with AIHA/Evans syndrome, the quantity of bone marrow CD5+ B lymphocytes showed significantly positive correlation with serum indirect bilirubin level (r = 1.00, P < 0.05). In Evans syndrome patients, the quantity of CD5+ B lymphocytes in bone marrow showed significantly positive correlation with platelet-associated immunoglobulin G (r = 0.761, P < 0.05) and platelet-associated immunoglobulin M ( r = 0.925, P < 0.05). The quantity of CD5+ B lymphocytes in bone marrow of all hemocytopenic patients showed significantly negative correlation with treatment response (tau-b = -0.289, P < 0.05) , but had no correlation with colony forming unit-erythroid (r = -0.205, P > 0.05) or colony forming unit-granulocyte-macrophage colonies (r = -0.214, P > 0.05)., Conclusions: The quantity of bone marrow CD5+ B lymphocytes in the patients with autoimmune hemocytopenia significantly increases and is correlated with disease severity and clinical response, which suggest that CD5+ B lymphocytes might play an important role in the pathogenesis of autoimmune hemocytopenia.

Two experiments were used to examine the potential role of IFN-gamma in chickens infected with reticuloendotheliosis virus (REV) and Marek's disease virus (MDV). First, chickens were infected with REV and/or MDV at 5 days of age and examined from 3 to 50 days post-infection (dpi). In REV+MDV co-infection chickens, IFN-gamma ELISA demonstrated a 3-fold increase at 7 dpi compared to the controls, while REV alone caused a 5-fold increase, the IFN-gamma levels peaked, and then gradually decreased. IFN-gamma levels significantly decreased in MDV infection at 3 dpi and 15 dpi. Second, experiments were designed to determine the effects of different viruses and ConA on IFN-gamma production. For REV- or MDV-infected chickens, the IFN-gamma levels decreased slightly after adding ConA. This is the first report of IFN-gamma production in SPF chickens infected with REV and MDV measured by directly quantitative method.

Twenty-four isolates of Newcastle disease virus (NDV) prevailing during 1997 -- 2005 in China were collected. These isolates were purified by CEF plaque assay and replicated in SPF chicken embryos. The hemagglutinin-neuraminidase (HN) genes of these viruses were cloned and sequenced. The HN gene sequences of thirty-six NDV reference strains in GenBank were also used in this study. The amino acid homologing of these viruses were compared and analyzed. The correlations among different fragments of HN gene were also analyzed. The results indicated that the homology of Chinese field NDV strains was 94.4%-99.4%, but 86.9%-89% compared with LaSota and Clone30, 87.9%-89.9% to F48E9, and 87.2%-96.2% to foreign NDV strains. There had the nearest distances among Chinese NDV isolates as compared with that of the LaSota, Clone30 and F48E9 by the phylogenetic tree. However, the distances of seven foreign NDV isolates were very close to Chinese NDV isolates as compared with these of the other foreign NDV isolates. We also found that all the Chinese field isolates were devoid of glycosylation site in position 538 -- 540. There were good correlations between different length amino acid fragments and the genomes of HN, especially the 5'-terminus first 80aa.

Newcastle disease virus (NDV) field strain SQZ04 was isolated from a broiler flock with typical symptoms and lesions, and cloned by plaque-purification three times. NDV SQZ04 was determined as a virulent strain with MDT of 50.5h and 51.2h, ICPI of 2.0 and 1.92, IVPI of 2.8 and 2.68 respectively before and after plaque-purification. Analysis of F gene indicated that SQZO4 was determined as a virulent gene type II , and its protein amino acid sequence has homologies of 99.3% , 98.7% and 96.9% with published gene type II vaccine strains LaSota, B1, virulent strain Taxas48,much higher than homologies of 88.3% - 88.6% or 91.3% - 92.1% with published gene types VI and IX. This is the first virulent field strain of gene type UI reported in China. Further more, the amino acid sequence 111 GGRQGRL117 in the F protein cleavage site in SQZ04 strain is identical to lentogenic strains of NDV, such as vaccine strains LaSota, B1. This is the first report that virulent NDV could have lentogenic amino acid sequence in the cleavage site of F protein, where HN genes was compared SQZ04 has a higher homologies of 95.3%- 97.3% with known velogenic strains, but lower homologies of 87.8% - 89.5% with published lentogenic strains.

Objective: To investigate the quantities of monocyte-derived dendritic cell precursors (pDC1) and plasmacytoid dendritic cell precursors (pDC2) in peripheral blood mononuclear cells (PBMC) of severe aplastic anemia (SAA) patients before and after immune suppressive therapy (IST), the ratio of the pDC1 to pDC2, and the expression of co-stimulating molecules (CD80, CD86, CD40) on dendritic cells (DC) and B cells in SAA patients., Methods: By means of three color monoclonal antibody labeling technology, the quantities and ratio of pDC1 and pDC2 in PBMC were detected in 26 SAA patients at active phase, 13 at recovery phase and 15 normal controls respectively. The aforementioned parameters of 10 SAA patients were tested before and 2 months after IST. The expression of CD80, CD86 and CD40 on DC and B lymphocytes were detected in 16 SAA patients and 15 normal controls., Results: The percentages of pDC1 and the ratio of pDC1/pDC2 of controls were (0.41 +/- 0.05)% and 1.58 +/- 0.18 respectively, and those of SAA patients at active phase were (0.67 +/- 0.13)% and 2.70 +/- 0.32 respectively, [pDC1 (P < 0.05); pDC1/ pDC2 ratio (P < 0.01)]. The aforementioned parameters in convalescent SAA patients decreased to (0.43 +/- 0.10)%, and 1.78 +/- 0.36 respectively, being no difference from those of normal controls. The percentages of pDC1 and pDC2 in 10 SAA patients were (0.87 +/- 0.31)%, and (0.35 +/- 0.09)%, before IST, and (0.24 +/- 0.09)%, (0.14 +/- 0.04)%, after IST, being significantly decreased (P < 0.05). The percentages of CD86 expression on DC of controls was (11.97 +/- 4.31)%, and that of SAA patients was (29.84 +/- 3.02) % (P < 0.05). The percentages of CD80, CD40 and CD86 expression on lymphocytes of controls were (2.57 +/- 0.44)%, (7.34 +/- 1.22)% and (1.86 +/- 1.11)%, respectively, and those of SAA patients were (5.17 +/- 0.68)%, (8.85 +/- 2.94)% and (5.98 +/- 0.96)% respectively (P < 0.05, P < 0.01). The percentage of CD86 expression on B lymphocytes in controls was 8.04 +/- 0.66%, and in SAA patients was (20.46 +/- 2.78)%, (P < 0.05)., Conclusion: The pDC subtypes were abnormal and the percentage of pDC1 is increased in SAA patients, which are associated with stage of this disease. DC and B Lymphocytes in SAA patients upregulated expression of costimulatory molecules (CD86) which cause the T lymphocyte abnormally activated.

By use of genomic DNA prepared from 5 fowlpox virus (FPV) vaccines made in China (from Shandong, Beijing, Liaoning, Zhejing and Shanghai respectively) as the templates, reticuloendotheliosis virus (REV) LTR was amplified in PCR with a pair of primers synthesized according to the sequences flanking the integrated REV-5'LTR in FPV genomes published in US and Australia. Sequence analysis indicated that the REV-5'LTR integration sites in genomes of all 5 Chinese PFV vaccine products were identical to American and Australian FPV vaccines with integrated REV-5' LTR. Among the 5 FPV vaccine products made in China, 3 productes Vac-B-Ch, Vac-D-Ch and Vac-E-Ch had the integrated REV-5' LTR sequences of 223bp with 100% homology to that in American vaccine Vac-3-Am and Australian vaccine Vac-M3-Au. The integrated 505bp REV-LTR sequences in another Chinese products Vac-A-Ch and Vac-C-Ch had 99.6% homology to the integrated REV-LTR of American vaccine Vac-1-Am and Australian vaccine Vac-S-Au. However, REV-5'LTR integrated in all 5 FPV vaccines made in China gave only 75.4%-91.5% homology to LTR of a Chinese field strain HA9901 of REV. Based on the above results, it is reasonable to speculate that the virus stocks for the 5 Chinese FPV vaccine products were rather originally imported with their integrated REV LTR than recombinated with LTR of REV local field strains in China.

Objective: To investigate the role of the burden of abnormal hematopoietic clone in the development of myelodysplastic syndromes (MDS)., Methods: The ratio of the bone marrow cells with abnormal chromosomes to the total counted bone marrow cells was regarded as the index of MDS clone burden. The disease severity related parameters including white blood cell count, hemoglobin, platelet count, lactate dehydrogenase level, bone marrow blast, myeloid differentiation index, micromegakaryocyte, transfusion, interleukin-2, tumor necrosis factor (TNF), CD4+ and CD8+ T cells of MDS patients were assayed, and the correlations between those parameters and MDS clone burden were also analyzed., Results: The clone burden of MDS patients was 67.4% +/- 36.2%. MDS clone burden positively correlated with bone marrow blasts (r = 0.483, P < 0.05), negatively with hemoglobin level (r = -0.445, P < 0.05). The number of blasts, hemoglobin, and erythrocytes in high clone burden (> 50%) and low clone burden ( < or = 50%) groups were 7.78% +/- 5.51% and 3.45% +/- 3.34%, 56.06 +/- 14.28 g/L and 76.40 +/- 24.44 g/L, (1.82 +/- 0.48) x 10(12)/L and (2.32 +/- 0.66) x 10(12)/L, respectively (all P < 0.05). CD4+ T lymphocytes of MDS patients and normal controls were (0.274 +/- 0.719) x 10(9)/L and (0.455 +/- 0.206) x 10(9)/L, respectively (P < 0.05). CD8+ T lymphocytes of MDS patients and normal controls were (0.240 +/- 0.150) x 10(9)/L and (0.305 +/- 0.145) x 10(9)/L, respectively. The serum level of interleukin-2 of MDS patients (6.29 +/- 3.58 ng/mL) was significantly higher than normal control (3.11 +/- 1.40 ng/mL, P < 0.05). The serum level of TNF of MDS patients and normal control group were 2.42 +/- 1.79 ng/mL and 1.68 +/- 0.69 ng/mL, respectively. The ratio of CD4 to CD8 was higher in high clone burden MDS patients (1.90 +/- 0.52) than that in low clone burden patients (0.97 +/- 0.44, P < 0.05)., Conclusion: The quantitive clonal karyotype abnormalities and deficient T cell immunity are important parameters for evaluating MDS severity and predicting its progression.

Prevailed Newcastle disease virus isolates were collected during 1999-2005 in China. These isolates were purified by CEF plaque assay and replicated in SPF embryos. The fusion protein (F) gene and hemagglutinin-neuraminidase (HN) gene of these isolated viruses were cloned and sequenced. Some of the F gene and HN gene sequences from GenBank were also used in this study. The homologies of nucleotide and amino acids were compared and correlations were analyzed by SPSS8.0 software among different length sequences of the F gene or HN gene. The nucleotide homologies and correlation among the F gene and HN gene were also analyzed. The results indicated there are good correlation among different length sequences of the F gene or HN gene and the F genome or HN genome (r > or = 0.973). There was also good correlations among different length amino acids of NDV F protein or HN protein (0.911< or = r < or = 0.968). But, there was only a less correlation between the whole F gene and HN gene (r = 0.312). The heredity mutation of HN genes had the character of geographical areas. The sequences of HN gene in Chinese isolates had an identity of more than 97%. But there was only 79.2% - 80.7% in HN nucleotide homology among the Chinese isolates and La Sota (vaccine).

Objective: Dingchuantang is a traditional Chinese medicine decoction which has the Qing, Xuan and Jiang components which have the ability to dispel (Xuan), depress (Jiang) and heat clear or febrigugal (Qing). It has been used for asthma and respiratory syncytial virus (RSV) infection. However, the therapeutic mechanisms of the decoction have not been identified. To understand the mechanism of the anti-RSV property of Dingchuantang, this study investigated the effects of Dingchuantang and its three components on the levels of interleukin-4 (IL-4) and interferon-gamma (gamma-IFN) in mice infected by RSV., Methods: One hundred and twenty mice were randomly assigned into Normal, Infected, Dingchuantang-treated, Qing component-treated, Xuan component-treated and Jiang component-treated groups (n=20 each). The RSV infection model was prepared in the rats from the last 5 groups. The 4 treatment groups were administered with Dingchuantang, Qing component, Xuan component and Jiang component by gastric lavage respectively 2 hrs after RSV innoculation. The contents of RSV in the lungs were tested by reverse transcription polymerase chain reaction (RT-PCR). The levels of IL-4 and gamma-IFN in broncho-alveolar lavage fluid (BALF) were measured by enzyme linked immunosorbent assay (ELISA)., Results: The contents of RSV of lungs in the Dingchuantang-treated group, and Qing and Xuan component-treated groups were significantly lower than those in the untreated Infected group. There was no significant difference between the Jiang component-treated and the Infected group. Dingchuantang treatment decreased the IL-4 level and increased the gamma-IFN level. The Xuan and Jiang component treatment decreased the IL-4 level but had no effect on the gamma-IFN level, while the Qing component treatment increased the gamma-IFN level but had no effect on the IL-4 level., Conclusions: Dingchuantang can inhibit RSV reproduction and this effect works by Qing and Xuan components. Dingchuantang can also restore the imbalance TH2/TH1 by depressing IL-4 levels, worked by Xuan and Jiang components, and increasing gamma-IFN levels, worked by Qing component.