At present ,the mechanism of highly pathogenic avian influenza H5N1 virus causing human infection or death is still not fully clear .In order to better understand the pathogenesis of the disease ,the rhesus macaques were infected with H5N1 virus (AF148678/ACGoose/Guangdong/11961H5N1) .We analyzed the clinical symptoms ,characteristics of the virus invades body ,pathological changes ,and immune response to discuss the pathogenesis of viral pneumonia induced by H 5N1 virus infection from the early time to the recovery time .The rhesus macaques were infected with H5N1 virus through nasal .Clinical signs were assessed daily ,and major organs and blood were collected for detection of blood routine analysis ,viruses were isola-ted and titrated from organs ,and pathologic and immunohistochemical were also conducted .As a result ,the rhesus macaques in-fected with H5N1 virus experienced fever ,dyspnea ,and anorexia .The respiratory tract was the major target of the virus and the virus could not replicate in organs outside the respiratory tract .Positive staining cells by immunohistochemistry were bronchial epithelial cells and alveolar macrophages .Rhesus macaques experienced temporary severe pneumonia after 1-3 days ,mainly be-cause of neutrophils infiltration ;gradual recovery 6 days later ,mainly with macrophage infiltration ;lung tissue presented recov-ery state after 14 days ,mainly with T lymphocytes infiltration .Finally ,we concluded that the predilection of the H 5N1 virus to infect the lower airway suggests that it may be a limiting factor in human-to-human transmissibility of the H5N1 virus .The pathogenesis may include virus invasion ,replication and immune injury .

Objective To study the cell immunity and eytokines responses to avian influenza A H5N1 virus infections in a BALB/c model to better understand the pathogenesis of H5N1 avian influenza disease. Methods Two hundred and twenty BALB/c mice of the infected group were inoculated with 0.1 ml (10-4.875 TCID50) of A/Goose/Guangdong/NH/2003 ( H5N1 ) virus intra-nasally. Fifty control mice received noninfectious allantoic fluid and another fifty control mice received normal sodium. Blood and spleen samples were collected from the live mice every 24 h during the 14 d post-infection. The changes of CD3 + T cells , CD4 + T cells, CD8 + T cells for cell immunity in blood circulation and spleen were detected by flow cytometry. And the cytokines and antibody responses in blood circulation were detected by ELISA. Necropsy was performed on mice that died during the experiment and those euthanized at end of study. Results Avian influenza A( H5N1) virus infections can make damages to the cell immune system transiently. The CD3 + T cells, CD4 + T cells, CDS + T cells declined at 24 days post infection in blood circulation and declined at 5-8 days in spleen, then recovered to the normal level gradually. The eytokines responses to the infections can be detected: the level of IFN-γ,TNF-α declined, IL-4, IL-18, IL-10 increased, and IL-2 changed little. The antibody increased rapidly from day 7 post infection until the end of the study (day 14 post infection). Conclusion Collectively, avian influenza A(H5N1) virus can cause cell immunity deficiency and an imbalance in the level of eytokines, which may contribute to the unusual severity of disease caused by the H5N1 avian influenza virus.

Objective To test our hypothesis that sensitivity to avian influenza A(H5N1)virus varies among mouse strain backgrounds, we compared the pathogenicity of H5N1 viral infection in 5 mouse strains. Methods Onehundred-fifty mice from 2 inbred strains(BALB/c and C57BL/6), and 3 outbred stocks(ICR, NIH Swiss, and KM Swiss)were used. Thirty mice of each strain were subjected to an infected group(20 mice), in which mice were inoculated with 0. 1 mL(104.875 TCID50)of A/Goose/Guangdong/NH/2003(H5N1)virus intra-nasally; ten control mice received noninfectious allantoic fluid. Clinical signs were assessed daily for 14 days post-infection. Necropsy was performed on mice that died during the experiment and those euthanized at end of study. Tissue samples were collected for viral isolation and pathological analysis. Results H5N1 virus infection can cause respiratory illness in all 5 strains with severe or minor acute respiratory distress symptoms, but with different mortality rates: 70% in BALB/c; 50% in ICR; 40% in NIH Swiss; 25% in C57BL/6; and 10% in KM Swiss mice. Necrotizing interstitial pneumonia was found in all cases of death. The virus was isolated from the lungs of all infected dead mice. Conclusion A/Goose/Guangdong/NH/2003 (H5N1)virus can infect all mouse strains used in this study, and can cause clinical symptoms and pathological changes similar to those found in humans infected with HSN1 viruses. However, the pathogenicity of H5N1 viral infection varies significantly between the different mouse strains. Thus, in future study of H5N1 virus infections the mouse strain most relevant to their particular research purpose should be selected as animal model.

Objective To investigate the expression levels of serum miR-210and miR-375in patients with non-small cell lung cancer (NSCLC).Methods A total of 25NSCLC patients (NSCLC group) and 14healthy volunteers (control group) were enrolled in this study.The relative expression levels of 9miRNAs (miR-182, miR-126, miR-31, miR-21, miR-221, miR-200b, miR-183, miR-210and miR-375) in 6 NSCLC patients and 6healthy volunteers were measured by RT-qPCR.The dysregulated miRNAs will be selected as candidate miR-NAs.The diagnostic value were evaluated by ROC curve.Results Compared with control group, 2 (miR-210and miR-375) out of 9miRNAs were up-regulated in NSCLC group, and the differences were statistically significant (P＜0.05), while the other 7miRNAs were not consistent with the reported literatures.Therefore, miR-210and miR-375were selected as candidate miRNAs.We found that the relative expression level of miR-210in the lung adenocarcinoma group was significantly different from control group (P＜0.05), while there was no significant difference between the squamous cell carcinoma group and the control group (P＞0.05).There was no significantly statistical difference in the relative expression level of miR-375between lung squamous cell carcinoma group, lung adenocarcinoma group and the control group (P＞0.05).The AUC of serum miR-210of lung adenocarcinoma group was 0.737 5 (95％CI:0.498 3-0.976 7, P=0.091 4) with a medium diagnostic value.Conclusion MiR-210is highly expressed in the serum of patients with lung adenocarcinoma, suggesting that miR-210may be a novel tumor marker for the diagnosis of lung adenocarcinoma.The value of miR-375in the diagnosis of lung cancer still needs to be further explored.

Objective:To explore arboviruses carried by midges in Shizong county, Yunnan province.Methods:A total of 74 batches of 3705 midges collected from Shizong county, Yunnan province in July 2013 were detected by RT-PCR method, and then the amplified bands were cloned and sequenced. MegAlign in DNAStar software was used for homology analysis, MEGAX software ALIGN for sequence alignment and genetic evolution analysis based on Maximum Likelihood (ML) method .Results:Among 74 batches of midge samples collected in Yunnan province, 600 bp electrophoresis bands were amplified in 3 batches: SZC33, SZC42 and SZ625C8-2, while no electrophoresis bands were amplified in the other 71 batches of midge samples. After cloning, we selected 5 clones for sequencing, among which the 5 clones of sample SZC42 obtained 589 nt sequences. BLASTx comparison showed that the 5 cloned sequences of SZC42 had the highest amino acid homology(69%) with the non-structural protein gene of Culex bastrovirus-like(CAVL) virus found in mosquitoes collected in the United States, which was designated as Midge bastrovirus-like virus (MAVL). The result of genetic evolution analysis showed that MAVL detected in midges formed a single evolutionary branch, and it’s homology of nucleotides and amino acids with other astrovirus(AV) was less than 71.4%. Further analysis revealed that MAVL had a close genetic evolution relationship with CAVL (NC_040647) detected in American mosquitoes, AV (MF042208) detected in Brazilian sewage and AV (NC_032426) detected from Vietnamese bats, and the homology was 61.4%-66.2% (nt) and 67.7%-71.4% (aa), while far genetic evolution and low homology of nucleotides and amino acids with other AV sirains.Conclusions:A new astrovirus (MAVL) was detected in midges collected from Shizong county, Yunnan province in 2013.

【Objective】 To explore the prognosis of critically ill patients with coagulation dysfunction using thrombelastogram(TEG) and coagulation four items combined with APACHEⅡ score. 【Methods】 From March 2017 to March 2020, 287 critically ill patients with coagulation dysfunction in our hospital were selected as the study group, and 303 patients with normal coagulation function during the same period were set as the control. The study group was divided into low-risk group(group A), intermediate-risk group(group B) and high-risk group (group C) based on the APACHEⅡ score, and into survival group and death group according to the prognosis. The difference of TEG, coagulation four items, and APACHEⅡ scores between the two groups were analyzed. The correlation and difference between TEG, coagulation four items and APACHE II score in the study group were analyzed. The ROC curve was drawn to analyze the prognostic predictive value of research indicators. 【Results】 Blood coagulation function related indicators in the study group fluctuated significantly: in comparison to the control, the CI value, MA value, and α angle were smaller, while the K time and R time were longer; among the coagulation four items, PT, APTT and TT were higher; Fg level was lower, and the APACHE Ⅱ score was higher(P<0.05). There were significant differences in relevant test index levels among patients with different degree of illness (APACHE Ⅱ score) in the study group. With the aggravation of the disease, the CI value, MA value, α angle, and TT continued to decrease, while K time, R time, APTT and PT showed a continuous increase trend (P<0.05). However, with the intensification of coagulation dysfunction, no significant increase or decrease was noticed in Fg(P>0.05). There were significant differences between the TEG and coagulation function related index levels in patients with different prognosis. Compared with the survivals, the CI value, MA value and α angle of the dead group were smaller, while the K time and R time were longer; and among the coagulation four items, PT, APTT, and TT were higher, the Fg level was lower, and the APACHEⅡ score was higher (P<0.05). The ROC curve showed that the corresponding AUC values of PT, APTT and INR were 0.701, 0.693 and 0.702, respectively, (P<0.05). The difference was statistically significant and had predictive value for death, but the accuracy was moderate.The combined indicators showed that the AUC values corresponding to APACHE Ⅱ score, P1, P2, P3, P4, P5, and P6 were 0.899, 0.751, 0.657, 0.759, 0.921, 0.921and 0.942, respectively, and the difference was statistically significant(P<0.05). The combined indicators have predictive value for death, except for P2<0.7, the rest were between 0.7~1.0, and the accuracy was P6>P4\\P5>APACHE Ⅱ score>P1>P2. 【Conclusion】 TEG, coagulation four items, and APACHE Ⅱ score can be used to assess the severity of patients with severe coagulation dysfunction. and the combined application of the 3 indicators are of high value in predicting the prognosis of such patients, and can provide reference for clinical formulation or adjustment of intervention programs to correct coagulation dysfunction.