A quail-origin subtype of the influenza virus was isolated from a human-infecting H7N9 subtype of the avian influenza virus found in a live poultry market and was given the name A/Quail/Hangzhou/1/ 2013 (H9N2). We analyzed the whole genome of this virus and its biologic characteristics. Sequence analyses suggested that the: HA and NS genes belonged to a CK/BJ/1/94-like lineage; NA, NP, PA and PB1 genes belonged to a SH/F/98-like lineage; M and PB2 genes belonged to a G1-like lineage. Analyses of key amino acids showed that the cleavage site in HA protein was PSRSSR ↓ GL, and that the HA protein had a human receptor-binding site with Leu226. Deletion of amino acids 69 - 73 was detected in the stalk of NA protein, the M2 protein had an Asn31 mutation, and the NS1 protein had two mutations at Ser42, Ala149. The intravenous pathogenicity of this virus was 0.36. A study in chickens suggested that all inoculated birds shed the virus from the trachea and cloaca on the third day post-infection (p. i. ) until 11 days. All chickens that had direct contact shed the virus on the second day p. i. until 8 days. Results of virus reisolation suggested that lung and tracheal tissues could shed the virus in 5 days, whereas the other organs could shed the virus in 3 days. These results suggest that this virus strain is H9N2 subtype LPAIV, whose lineage is prevalent in mainland China. This research provides evidence on how to monitor and prevent the H9N2 subtype of the avian influenza virus.
Animals ; Chick Embryo ; Chickens ; China ; Genotype ; Influenza A Virus, H9N2 Subtype ; classification ; genetics ; isolation & purification ; Influenza in Birds ; virology ; Phylogeny ; Quail ; virology

Objective To explore the behavior characteristics in children with benign epilepsy combined with centro-temporal spikes(BECTS).Methods Eighty-two children with BECTS aged 2.5-3.0(2.65?2.31)years old,51 male,31 female,who were free of mental retardation assessed with Gesell developmental schedules,untreated with antiepileptic drugs,and were investigated 15 days after the latest seizure.Eighty-two healthy children with sex and age matched to the cases,53 male,29 female,aged 2.5-3.0(2.6?0.4)years old.The behavior characteristics of infants in BECTS group and control group were assessed with CBCL,including 6 behavior factors which were sleep problem,social flinches,depression,physical aspect,attacking,act of sabotage and the infants-middle school student social ability scale.Results The total scores of behavior characteristics and the scores of depression,sleep problem,attacking and act of sabotage in BECTS group were all higher than those in control group,the differences were statistically significant.However,scores of social flinches,and physical aspect in BECTS group had no significant differences compared with those of control group.There were no significant difference of social adaptive component between the BECTS and control group.Conclusions Children with BECTS have behavior disorders to some extent,but their social adaptive capacity are the same as normal children.

PurposeIt is important to keep the consistency of CT perfusion imaging (CTP) measurement in the evaluation of intracranial disorders. The purpose of this paper is to probe into the consistency of basal ganglia hemodynamic characteristics in normal rabbits with CTP measurement in the mode of volume helical shuttle (VHS).Materials and Methods Brain CTP scans of 10 healthy rabbits were carried out in the VHS mode. The images derived from CTP were separately analyzed by two radiologists (A and B) with different experiences. ROIs were placed in the bilateral basal ganglia of the subjects and parameters such as cerebral blood volume (CBV), cerebral blood flow (CBF) and mean transit time (MTT) were measured three times in each rabbit. Thirty days later, radiologist A analyzed the CTP images again with the same method. Differences of parameters in bilateral basal ganglia measured by different radiologists at different periods were compared respectively and consistency of the measurements was further assessed.Results There was no significant difference among the three measurements of CBV, CBF and MTT in bilateral basal ganglia (F=0.162, 0.050 and 0.488,P>0.05). TheICC of CBV, CBF and MTT measured by the two radiologists for the first time were 0.84, 0.81 and 0.79; while theICC of CBV, CBF and MTT measured by radiologist A twice were 0.80, 0.89 and 0.84, which showed that the results had good consistency measured by different radiologists at different times.Conclusion With the same post-processing software, the consistency of CTP of basal ganglia in the VHS mode is satisfactory in healthy rabbits, which suggests this technique may be a suitable means in the evaluation of basal ganglia hemodynamics in animal models.

Cutaneous Fistula ; Digestive System Abnormalities ; therapy ; Endoscopy, Gastrointestinal ; methods ; Female ; Humans ; Infant ; Treatment Outcome

Aim TodevelopandvalidateaLC-MS/MS assay to quantify halometasone in rabbit plasma and study pharmacokinetics of halometasone after dermal topical administration of Halometasone Cream.Meth-ods Theplasmasamplewassubmittedtoliquid-liquid extraction using methyl tertiary butyl ether,with dexa-methasone as the internal standard (IS ).Chromato-graphic separations were performed on a Diamonsil C18 column(100 mm ×4. 6 mm,5 μm)with a linear gra-dient of methanol and 2 mmol · L-1 ammonium ace-tate.Halometasone and dexamethasone(IS)were ion-ized with an ESI source operated in negative ion mode, and the detected ions were m/z 503. 1→413. 0 (halo-metasone),m/z 391. 0→361. 0 (dexamethasone ). The test article could be monitored in rabbit plasma when following single dermal topical administration of Halometasone Cream at 1 g/100 cm2 to rabbits by u-singavalidatedLC-MS/MSassay.Results Calibra-tion curve was linear over the concentration range of0. 02~20 μg·L-1 in rabbit plasma.For low,medi-um,high concentration of QC solutions,the intra-and inter-day precision was in the range of 3. 72% ~7. 87%, and the accuracy was within 99. 1% to 103%. The pharmacokinetic parameters in rabbits were as follows:Tmax,Cmax,AUC0-t,T1/2 was (7. 38 ± 1. 06)h,(1. 16 ±0. 527)μg·L-1,(18. 8 ±7. 23)h·μg·L-1 ,(13. 8 ±3. 70)h,respectively.Conclusions ThisLC-MS/MSanalysismethodhashighsensitivi-ty,and sample processing method is simple,which has been rigorously validated.The method could be suc-cessfully applied to the pharmacokinetic study of halo-metasone after skin administration of Halometasone Cream to rabbits.

Background Researches demonstrated that CpG ODN,a immunostimulatory sequences,has preventing and treating effect on allergic conjunctivitis caused by protein allergen.However,its effect on allergic conjunctivitis caused by fungal allergen is unclear. Objective This study aimed to investigate into whether the Th1-Th2 switching immunostimulatory CpG ODN could reverse the response in the murine allergic conjunctivitis model caused by aspergillus fumigatus. Methods A mixture of spores and hyphae of aspergillus fumigatus strain was used to induce allergic conjunctivitis in male BALB/C mice aged 6-8 weeks.This experiment was designed into preventive or therapeutical treatment program.Under both settings,allergic conjunctivitis of the animals were treated with CpG ODN,nonstimulatory GpC ODN or PBS.After the last challenge with the allergen,the clinical symptoms of the animals were scored based on the criteria of Magone.The animals were sacrificed and the histopathological examination of conjunctiva was performed.Expression of TLR4 mRNA in conjunctiva was analyzed by real-time PCR assay.The responsiveness and populations of lymphocytes in spleen and draining lymph nodes were analyzed by flow cytometry.The use complied with the Standard of Association for Research in Vision and Ophthalmology. Results In the prevention mode.CpG ODN decreased subconjunctival infiltration compared with GpC ODN and PBS groups with the average neutrophil count index(21.25 ±11.59/section,30.75 ±11.44 section and 69.00±9.90/section,respectively).Expression of TLR4 mRNA was up-regulated significantly by CpG ODN.The clinical scores for CpG ODN group were insignificantly lower than those in GpC ODN group and PBS group(P＞0.05).In the therapeutic mode,compared with GpC ODN and PBS groups,the allergic symptom score in CpG ODN group manifested significantly lower(t=4.000.t=2.750,P＜0.01)and showed fewer cellular infiltration(t=4.870,t=3.829,P＜0.01)and higher expression of TLR4 mRNA(P＜0.01).In cultured splenic and draining lymph node cells,increased percentages of CD4+ CD25+ and CD4+ CD25+ CD69+ in CpG ODN group were observed compared with control groups(|P＜0.05). Conclusion CpG ODN can relieve aspergillus fumigatus-induced allergic conjunctivitis via either subconjunctival injection or topical application by upregulating expression of TLR4 and activating Treg lymphocytes.

Objective To investigate the expression of osteopontin and matrix metalloproteinase-9 (MMP9),and the relationship of osteopontin and MMP9 in malignant melanoma.Methods Expression of osteopontin and MMP9 was measured by immunohistochemical SP method in 23 patients with primary cuta- neous malignant melanoma,17 patients with metastatic melanoma and 20 patients with pigmented nevus. Results Osteopontin and MMP9 were expressed respectively in 87.5% and 75.0% of 40 malignant melanoma specimens,15.0% and 10.0% of 20 pigmented nevus specimens.The expression of both osteo- pontin and MMP9 was significantly higher (both P＜0.05) in malignant melanoma than in pigmented ne- vus.There was no correlation between the expression of osteopontin and MMP9,with age,sex,lymph node metastasis or location of lesions (P＞0.05).Twenty-nine cases were positive for both osteopontin and MMP9,4 negative for either osteopontin or MMP9.Conclusion Both osteopontin and MMP9 were over- expressed in malignant melanoma,but neither was related to lymph node metastasis.

Puerarin (PUE), as an isoflavone component, has a wide range of pharmacological activities, while its poorly aqueous solubility limits the development of solid oral dosage forms. In this study, PUE along with nicotinamide (NIC) were prepared into the coamorphous system by solvent-evaporation method and characterized by powder X-ray diffraction (PXRD), differential scanning calorimetry (DSC) and Fourier transform infrared spectroscopy (FT-IR). In addition, its dissolution behavior and solubilization mechanism were also investigated. PUE-NIC coamorphous was a single homogeneous binary system, with a single glass transition temperature at 35.1 ℃. In comparison to crystalline PUE, during the dissolution process, coamorphous PUE-NIC not only exhibited the "liquid-liquid phase separation" (LLPS) phenomenon, but the formation of A_p type complexation (1∶1 and 1∶2) between PUE and NIC molecules was also verified, which significantly improved the solubility of PUE and prolonged the supersaturation time, and would benefit its absorption.

Abstract：Programmed death receptor⁃ 1（PD⁃1）belongs to the family of immunoglobulin B7⁃CD28，which plays an important role in regulating immune response in human body. Since the first PD⁃1／PD⁃ligand 1（PD⁃L1）monoclonal antibody was approved for marketing in China in 2018，the value of PD⁃1／PD⁃L1 immunotherapy in oncotherapy has attracted wide attention. Based on the introduction of the action mechanism of PD⁃1／PD⁃L1 mAbs，this paper reviews the application progress of 8 on ⁃ market PD ⁃ 1／PD ⁃ L1 mAbs in China in oncotherapy from the perspectives of approved indications，clinical trials，usage and dosage，and adverse reactions，in order to provide reference for the rational appli⁃ cation of PD⁃1／PD⁃L1 monoclonal antibodies in clinic.

Objective To compare the effects of hepatic vein occlusion with tourniquet and Satinsky clamp in reseeting liver tumor involving the second hepatic portal.Methods From Jan 2003 to Jun 2006,180 patients underwent major liver resection with the selective hepatic vascular exclusion (SHVE).According to methods of hepatic vein occlusion,they were divided into two groups:Occlusion with tourniquet(tourniquet group,n=95)and occlusion with Satinsky clamp(Satinsky clamp group,n= 85).In tourniquet group,the hepatic veins were encircled and occluded with tourniquet,and in Satinsky clamp group,the hepatic veins were not encircled and clamped directly with Satinsky clamp.Data regarding the intraoperative and postoperative courses of the patients were analyzed.Results There was no difference between the two groups regarding the operating time,ischemia time,intraoperative blood loss and postoperative complications rate.The dissecting time of hepatic veins was significantly shorter in Satinsky group(6.2?2.4 min vs 18.3?6.2 min).lu the tourniquet group,five hepatic veins(one fight hepatic vein and four common trunk of left-middle hepatic veins)could not be dissected and encircled because of the tumors involving the cava hepatic junction.Another patient's common trunk of left-middle hepatic vein was inadvertently lacerated during the dissection.Hepatic veins in these 6 patients were occluded with Satinsky clamp successfully.Conclusion Occlusion with Satinsky clamping is safer and easier procedure than tourniquets in the resection of liver tumor involving the second porta hepatis.