In order to obtain polyclonal antibodies against the fibrillar(Fiber)protein of fowl ade-novirus serotype 11(FAdV-11)and investigate its cross-reactivity to different serotypes of FAdV Fiber,the gene encoding the FAdV-11 Fiber protein was cloned into a prokaryotic expression vec-tor pET-32a by homologous recombination technology,then the plasmid was transformed into BL21(DE3)receptor cells,and the purified recombinant protein was used as an immunogen to im-munize rabbits to prepare polyclonal antibody after induced expression,and the cross-reactivity of the polyclonal antibody against different serotypes of FAdV Fiber proteins was identified by West-ern blot and indirect immunofluorescence(IFA).The results showed that the His-FAdV-11-Fiber recombinant protein was mainly expressed as inclusion bodies and was well expressed.Western blot and IFA showed that the prepared polyclonal antibody reacted with the Fiber proteins of FAdV-8a,FAdV-8b,and FAdV-11,but did not with the 2 Fiber of FAdV-4(Fiber 1 and Fiber 2)proteins.In conclusion,in this study,we successfully prepared rabbit polyclonal antibodies against FAdV-11 Fiber and showed that it specifically recognized the Fiber proteins of FAdV-8a,FAdV-8b and FAdV-11,which lays the foundation for further establishment of serological differential diag-nosis of FAdV-11.

Breast cancer is the most common cancer in women and one of the deadliest cancers worldwide.According to the distribution of tumor tissue,breast cancer can be divided into invasive and non-invasive forms.The cancer cells in invasive breast cancer pass through the breast and through the immune system or systemic circulation to different parts of the body,forming metastatic breast cancer.Drug resistance and distant metastasis are the main causes of death from breast cancer.Research on breast cancer has attracted extensive attention from researchers.In vitro construction of tumor models by tissue engineering methods is a common tool for studying cancer mechanisms and anticancer drug screening.The tumor microenvironment consists of cancer cells and various types of stromal cells,including fibroblasts,endothelial cells,mesenchymal cells,and immune cells embedded in the extracellular matrix.The extracellular matrix contains fibrin proteins(such as types Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅵ,and Ⅹcollagen and elastin)and glycoproteins(such as proteoglycan,laminin,and fibronectin),which are involved in cell signaling and binding of growth factors.The current traditional two-dimensional(2D)tumor models are limited by the growth environment and often cannot accurately reproduce the heterogeneity and complexity of tumor tissues in vivo.Therefore,in recent years,research on three-dimensional(3D)tumor models has gradually increased,especially 3D bioprinting models with high precision and repeatability.Compared with a 2D model,the 3D environment can better simulate the complex extracellular matrix components and structures in the tumor microenvironment.Three-dimensional models are often used as a bridge between 2D cellular level experiments and animal experiments.Acellular matrix,gelatin,sodium alginate,and other natural materials are widely used in the construction of tumor models because of their excellent biocompatibility and non-immune rejection.Here,we review various natural scaffold materials and construction methods involved in 3D tissue-engineered tumor models,as a reference for research in the field of breast cancer.

OBJECTIVE: To explore the effect and mechanism of miR-125a-5p targeted regulation of scavenger receptor B1 (Scarb1) gene on anoxia/reoxygenation injury of rat cardiomyocytes. METHODS: H9c2 rat cardiomyocytes were randomly divided into blank control group, hypoxia/reoxygenation group, transfection control group and mir-125a-5p transfection group. The expression of miR-125a-5p, cardiomyocyte viability, apoptosis rate, ATP content and the expression of Scarb1, Cyt C, Bax, Bcl-2 and NF-κB signaling pathway related proteins were determined. Target gene of miR-125a-5p was predicted with Targetscan software, and the targeting of miR-125a-5p on Scarb1 was verified by double luciferase reporter gene experiment. RESULTS: Compared with the blank control group, the expression of miR-125a-5p, Bax, Cyt C and the apoptotic rate of cardiomyocytes in the hypoxia/reoxygenation group were significantly increased (P<0.05), while the expression of Scarb1, Bcl-2 and the content of ATP were significantly decreased (P<0.05). Compared with the control group, the situation of mir-125a-5p transfection group was just the opposite. Double luciferase reporter gene experiment has confirmed Scarb1 to be the target of miR-125a-5p. Hypoxia/reoxygenation can promote the expression of NF-κB p65, C-myc and Cyclin D1 in cardiomyocytes, while down-regulating the expression of miR-125a-5p can inhibit the expression of such proteins. CONCLUSION Hypoxia/reoxygenation can induce the expression of miR-125a-5p in rat cardiomyocytes. Inhibition of miR-125a-5p can protect cardiomyocytes from hypoxia/reoxygenation by up-regulating the expression of Scarb1. The mechanism may be related to the inhibition of activation of NF-κB signaling pathway.

Objective: To investigate the dynamic changes of antibodies induced by leptospiral vaccines. Methods: Antigens for antibody detection were screened out. ELISA was used to analyze antibody responses induced at different time points after immunizing guinea pigs with different batches of leptospiral vaccines from different manufacturers. To investigate the relationship between antibody responses induced by leptospiral vaccines and their protective effects in animal model, guinea pigs were challenged with Leptospira after immunization. Results: There was no significant antigen-antibody reaction between the LigA protein or Patoc Ⅰ antigen and the serum samples of guinea pigs immunized with leptospiral vaccines. Notable IgG and IgM antibody reactions were observed in all vaccination groups when using bacterial proteins from seven Leptospira reference strains which were used for the preparation of leptospiral vaccines as envelope antigens. Antigen-specific IgG antibodies peaked at 35 d after the last immunization, and the highest peak of antigen-specific IgM antibodies was reached 11 d after the last immunization. Results of the challenge test showed that non-diluted leptospiral vaccines induced significant IgG and IgM antibody reactions in guinea pigs as compared with those diluted three or nine times, showing good protective effects. Conclusions Analysis of the dynamic changes of antibodies induced by leptospiral vaccines revealed that there was correlation between the induced serum antibody responses and the protective effects. This study provided reference for further study on alternative methods for evaluating leptospiral vaccine potency.

Collaboration of c-KIT mutations with AML1-ETO (AE) has been demonstrated to induce t(8; 21) acute myeloid leukemia (AML). Targeted therapies designed to eliminate AE and c-KIT oncoproteins may facilitate effective treatment of t(8; 21) AML. Homoharringtonine (HHT) features activity against tumor cells harboring c-KIT mutations, whereas oridonin can induce t(8; 21) AML cell apoptosis and AE cleavage. Therefore, studies should explore the efficacy of combination therapy with oridonin and HHT in t(8; 21) AML. In this study, we investigated the synergistic effects and mechanism of oridonin combined with HHT in t(8; 21) AML cell line and mouse model. The two drugs synergistically inhibited cell viability and induced significant mitochondrial membrane potential loss and apoptosis. Oridonin and HHT induced significant downregulation of c-KIT and its downstream signaling pathways and promoted AE cleavage. HHT increased intracellular oridonin concentration by modulating the expressions of MRP1 and MDR1, thus enhancing the effects of oridonin. The combination of oridonin and HHT prolonged t(8; 21) leukemia mouse survival. In conclusion, oridonin and HHTexert synergistic effects against t(8; 21) leukemia in vivo and in vitro, thereby indicating that their combination may be an effective therapy for t(8; 21) leukemia.

Objective:To investigate the chemical constituents in the rattan of Rubia argyi L.. Methods:The air-dried rattan of Rubia argyi L. was powdered and extracted three times by 75% ethanol with refluxing. After removing the solvent under the reduced pressure,the crude extract was dissolved in water,and then filtrated and extracted by petroleum ether and ethyl acetate to obtain crude extract after removing petroleum ether and ethyl acetate. The compounds were isolated and purified by silica gel column chromatogra-phy,reversed-phase silica gel column chromatography and Sephadex LH-20 gel column chromatography,and then identified based on physicochemical properties and spectral analysis(1 H-NMR and 13C-NMR). Results:Totally 13 compounds were isolated from the rat-tan of Rubia argyi L.,and characterized as secoisolariciresinol(1),xanthopurpurin(2),daucosterol(3),dehydroabietic acid(4), 2-hydroxy-1-methoxy-anthraquinone(5),β-sitosterol(6),lirioresinol A(7),2-hydroxy-7-methyl-9,10-anthraquinone(8),strych-novoline (9), ciwujiatone (10), 3,4-divanillyltetrahydrofuran (11), 2-(4-hydroxypheny) -6-(3-methoxy-4-hydroxyphenyl)-3,7-dioxabicyclo[3,3,0]octane (12), and (6S,9R)-vomifoliol (13).Conclusion: The compounds 1-13 are isolated from the rattan of Rubia argyi L. for the first time and the compounds 1,2,4,5 and 7-13 are first isolated from Rubia L..

Aim To investigate the effects of dopamine(DA)on insulin secretion from rat islets and the possible mechanism.Methods Pancreatic islets were obtained from the pancreatic of male SD rats by collagenase P digestion and histopaque-1077 density gradient separation.Insulin secretion experiment was used to observe the change of insulin release after DA treatments.As to study the potential mechanisms of the effects of DA,patch-clamp experiment and calcuim image technique were applied to test the depolarization-evoked Ca2+ currents,action potential duration and intracellular Ca2+ concentration.Results In 2.8 mmol·L-1 glucose,DA had no effect on insulin secretion;in 16.7 mmol·L-1 glucose,dopamine inhibited insulin secretion in a dose-dependent manner.DA inhibited the inward calcium current,shorten the action potential duration,and reduced the intracellular Ca2+ concentration.Conclusion DA inhibits insulin secretion maybe by decreasing the inward calcium current leading to shorten the action potential duration and reduce the intracellular Ca2+ concentration.

OBJECTIVE:To investigate the characteristics and causes of cardiac ADR induced by quinolones,and to provide reference for clinical treatment. METHODS:Three thousaud two hundred and eighty-eighe 8 patients receiving common quinolones were selected from clinical departments in Zhengzhou Central Hospital of Zhengzhou University during Mar. 2012-Mar. 2016. Retro-spective analysis was conducted in terms of patients’age and gender,clinical departments,main clinical manifestations,route of administration,underlying disease,drug combination. The reasons for cardiac ADR were analyzed. RESULTS:Among 3 288 pa-tients,there were 34 patients(1.03%)with cardiac ADR. Among them,the incidence of cardiac ADR in patients over 50 years old was as high as 76.47%;patients with cardiac ADR were mainly in the respiration department,gastroenterology department and urology department,accounting for 76.47%;most of patients were from gastroenterology department(29.41%). In cardiac ADR, the main clinical manifestations were QTc interval prolongation torsadesde poiutes(TdP)and TdP,accounting for 58.82%. Among them,patients with QTc interval prolongation TdP occupied the highest proportion,there was no statistical significance compared to TdP(P>0.05);there was statistical significance in the difference with other clinical manifestations(P<0.05). Among common-ly used quinolones,levofloxacin(32.35%)and ciprofloxacin(41.18%)caused large proportion of cardiac ADR,there was statisti-cal significance compared to norfloxacin,moxifloxacin and other quinolones(P<0.05). The proportion of cardiac ADR induced by intravenous dripping(91.18%)was much higher than oral administration(8.82%),with statistical significance(P<0.05). Among patients with cardiac ADR,the patients with underlying disease (94.12%) and drug combination (91.18%) occupied the higher proportion,there was statistical significance compared to the patients without underlying disease and drug combination(P<0.01). Among drug combination,the patients receiving amiodarone (29.41%) and salbutamol (20.59%) occupied the large proportion, there was statistical significance compared to other types of drug combination(P<0.05). CONCLUSIONS:Cardiac ADR induced by quinolones in our hospital mostly occurs in respiration department,gastroenterology department and urology department,and mainly manifests as QTc interval prolongation TdP and TdP. The incidence of cardiac ADR may be greatly increased in elderly pa-tients,patients with underlying diseases,and drug combination as well as intravenous infusion. Therefore,clinicians should select suitable quinolones,and make reasonable individualized dosage regimen.

Objective To find out whether conversation analysis helps to differentiate psychogenic nonepileptic seizure (PNES) from epileptic seizure in Chinese patients.Methods Twelve unselected patients from Peking Union Medical College Hospital during 2014 to 2016 with diagnostic uncertainty were included.Interactions following standard protocol were carried out.A linguist blinded to all medical data and a neurologist studied videos and transcripts of the interactions.Using a diagnostic scoring aid which includes 17 conversation features summarized from previous researches, they attempted to predict the medical diagnosis of those patients independently.Results Accurate diagnosis was predicted in 10/12 patients by both raters.Average scores of patients with epileptic seizures were 8.00 (linguist) and 6.75 (neurologist), while average scores of paitents with PNES were-5.75 (linguist) and-7.88 (neurologist).Both raters agreed on most individual items (81.86%, 167/204).To demonstrate different features between these two groups, a case comparison was made between one patient with frontal lobe epilepsy and one patient with PNES.Conclusion In Chinese patients, conversation analysis can help differentiate between epileptic seizure and PNES.

Objective To analyze the correlation among 3 kinds of detection methods of routine microscopic examination,fluorescence PCR nucleic acid amplification and fungal color culture in the fungal detection of vaginal secretion.Methods The patients with suspected vaginitis treated in this hospital during 2014-2016 were selected.Each 500 cases of negative and positive vaginal secretion samples by microscopic examination were collected.The candida types were identified by using the fluorescence PCR nucleic acid amplification,then 100 samples with the positive results of fluorescence PCR nucleic acid amplification for detecting fungal were performed the fungal microbial culture to verify the accuracy rate of typing results.Results The Kappa value of consistency test between fluorescence PCR nucleic acid amplification and routine microscopic examination was 0.632,the consistency between them was poor.Among 500 positive samples of vaginal secretion detected by routine microscopic examination,382 cases (76.4 ％) of Candida albicans infection were detected by fluorescence PCR nucleic acid amplification,73 cases (14.6％) were Candida glabrata infection,10 cases (2.0 ％) were Candida tropicalis infection,3 cases (0.6 ％) were Candida albicans combined Candida glabrata infection and 32 cases (6.4 ％) were other fungal infection.Among 500 negative samples by conventional microscopic examination,152 positive cases were identified by fluorescence PCR nucleic acid amplification,including 130 cases of Candida albicans,16 cases of Candida glabrata and 6 cases of Candida tropicalis.There was no statistical significant difference in positive rate between the fluorescence PCR nucleic acid amplification and CHROMAgar rapid color method (x2 =0.131,P =0.936).Conclusion For the patients with clinical manifestations and negative microscopic examination results,it is recommended to conduct fluorescence PCR nucleic acid amplification rapid type identification or fungal culture identification.