The Ebola virus is highly infectious and can result in death in ≤ 90% of infected subjects. Detection of the Ebola virus and diagnosis of infection are extremely important for epidemic control. Presently, Chinese laboratories detect the nucleic acids of the Ebola virus by real-time reverse transcription-polymerase chain reaction (RT-PCR). However, such detection takes a relatively long time and necessitates skilled personnel and expensive equipment. Enzyme-linked immunosorbent assay (ELISA) of serum is simple, easy to operate, and can be used to ascertain if a patient is infected with the Ebola virus as well as the degree of infection. Hence, ELISA can be used in epidemiological investigations and is a strong complement to detection of nucleic acids. Cases of Ebola hemorrhagic fever have not been documented in China, so quality-control material for positive serology is needed. Construction and expression of human-mouse chimeric antibodies against the nucleoprotein of the Ebola virus was carried out. Genes encoding variable heavy (VH) and variable light (VL) chains were extracted and amplified from murine hybridoma cells. Genes encoding the VH and VL chains of monoclonal antibodies were amplified by RT-PCR. According to sequence analyses, a primer was designed to amplify functional sequences relative to VH and VL chain. The eukaryotic expression vector HL51-14 carrying some human antibody heavy chain- and light chain-constant regions was used. IgG antibodies were obtained by transient transfection of 293T cells. Subsequently, immunological detection and immunological identification were identified by ELISA, immunofluorescence assay, and western blotting. These results showed that we constructed and purified two human- mouse chimeric antibodies.
Animals ; Antibodies, Monoclonal ; genetics ; immunology ; Cloning, Molecular ; Ebolavirus ; genetics ; immunology ; Hemorrhagic Fever, Ebola ; immunology ; virology ; Humans ; Immunoglobulin Heavy Chains ; genetics ; immunology ; Mice ; Nucleoproteins ; genetics ; immunology ; Viral Proteins ; genetics ; immunology

Objective To investigate the effects of mefformin on the differentiation of osteoclastas well as relative mechanism.Methods Raw264.7 cells from the murine macrophage cell line was used.Receptor activator of NF-κB ligand (RANKL) was used to stimulate osteoclast differentiation from Raw264.7 cells.Osteoclast differentiation was assessed by tartrate-resistant acid phosphatase (TRAP) and actin fluorescence staining and counting the TRAP-positive cells after exposure to different concentrations of mefformin (0 μmol/L,400 μmol/L,800 μmol/L and 1000 μmol/L) or rapamicin (100 nmol/L) in the presence of 50 ng/ml RANKL for 5 days.Bone-resorbing activity was evaluated by BD BioCoatTM OsteologicTM Bone Cell Culture System.The expression of osteoclast-specific genes like TRAP,capthesin K,calcitonin receptor (CTR) and matrix metalloproteinase (MMP-9) was evaluated by RT-PCR.The expression of tumor necrosis factor-α(TNF-ct) S6K1Thr389,S6 Ser235/236,4E-BP1Thr37/46 and c-Fos protein was evaluated by ELISA kit and Western blot analysis,respectively.Results Mefformin dose-dependently inhibited RANKL-stimulated osteoclasts differentiation in Raw264.7 cell culture,as manifested by decrease of TRAP-positive multinucleated cells and pit erosion area,down-regulation of TRAP,cathepsin K,CTR and MMP-9 mRNA and reduction of TNF-α and c-Fos protein expression.Further study revealed that RANKL activated mTOR complex 1(mTORC1) signaling,while mefformin impaired RANKL-stimulated mTORC1 signaling.Rapamycin,an mTORCl-specific inhibitor and immunosuppressive macrolides could also prevent RANKL-induced osteoclast differentiation and bone resorption in vitro.Conclusion Mefformin inhibits osteoclastogenesis in vitro,which may due to reduction of TNF-α and c-Fos protein expression,and mTORC1 signaling is involved in this process.

Action Potentials ; Animals ; Ear, Inner ; blood supply ; Facial Nerve ; physiopathology ; Hearing Loss, Sensorineural ; etiology ; Rabbits

Viral infection is the main death cause of infectious diseases in China. The establishment of an animal model to mimic the progression of viral infectious diseases in humans is of great significance to the study of pathogenesis and prevention of viral infectious diseases. As a new animal model established and developed in recent years, tree shrew has showed obvious advantages and potentials compared with other non-human primates and mice which are commonly used as virus-infected animal models. In this paper, the biological advantages of tree shrew as a novel animal model of viral infectious diseases are summarized, including taxonomy, physiology and immunology. In addition, the latest application of tree shrew in the research of many viral infectious diseases such as hepatitis virus, herpes simplex virus, influenza virus and enterovirus infections are compared and summarized.

Objective: To investigate the comprehensive nutritional status and diet behavior of middle aged and elderly women with osteoporosis, and thereby to explore the relationship between diet behavior and comprehensive nutritional status. Methods: 311 middle-aged and elderly women with osteoporosis in Chengdu were included in this study. Mini Nutritional Assessment (MNA) was applied to assess their comprehensive nutritional status. Information of social-demographic characteristics and diet behavior (about meals, snacks and water drinking, etc.) of the subjects was collected by questionnaire. Chi square test was used to assess the differences in nutritional status among patients who have different eating behaviors. Logistic regression analysis was performed to evaluate the relationship between diet behaviors and comprehensive nutritional status. Results: The mean MNA score of subjects was 25.8±2.5. 20.3% (63/311) of the subjets were at risk of potential malnutrition, but there was no malnourished subjects found. 46.9% (46/311) of the subjects were in good appetite. 95.2% (296/311) of them had a fixed food intake each meal. 65.8% (198/311) of them had snacks every day, and the most common choice was fruit (86.4% (248/287)). 54.8% (165/311) of them had initiative drinking water habits, and the most common choice was plain boiled water (79.9%, 246/308). 76.5% (238/311) of them had daily portable water less than 1 500 ml. After adjusting the effects of age, occupation and education level, bad appetite (OR=3.50, 95%CI: 1.18-10.62), unfixed food intake (OR=7.27, 95%CI: 1.40-35.83), and seldom or never intake of snack (OR=3.71, 95%CI: 1.42-9.72) were risk factors for malnutrition risk, while tea drinking was protective factor(OR=0.31, 95%CI: 0.11-0.93). Conclusion Risk of potential malnutrition and unhealthy diet behavior among the middle aged and elderly women with osteoporosis should be paid more attention. Unhealtghy diet behavior has a negative effect on their comprehensive nutritional status.

OBJECTIVETo investigate the antioxidant and cytotoxic properties of five diarylheptanoids (1-5) isolated from the rhizomes of Zingiber officinale.

METHODVarious models such as scavenging superoxide anions and 1,1-diphenyl-2- picrylhydrazyl (DPPH) radicals, inhibiting lipid peroxidation, as well as protecting of rat pheochromocytoma (PC12) cells induced by hydrogen peroxide (H2O2) were employed to assay the antioxidative effects of the diarylheptanoids. The cytotoxicities of compounds 1-5 were measured with MTT assays.

RESULTThe test compounds (1-5) showed promising DPPH inhibitory activities, and compound 5 exhibited the strongest DPPH scavenging activity with an IC50 value of (22.6+/-2.4) micromol x L(-1). Compounds 1, 3 and 4 showed potential anti-peroxidative effects with inhibitory rates of (66.3+/-15.4)%, (68.7+/-15.8)% and (72.2+/-10.6)%, respectively, at 100 microg x mL(-1). It could be observed that compounds 1, 3 and 4 demonstrated significant neuroprotective activities in a dose-dependent manner. Moreover, compound 3 exhibited certain cytotoxicities against human chronic myelogenous leukemia cells (K562) and its adriamycin-resistant cells (K562/ADR) with IC50 values of (34.9+/-0.6), (50.6+/-23.5) micromol x L(-1), respectively.

CONCLUSIONIn vitro results demonstrated that five diarylheptanoids (1-5) isolated from the roots of Z. officinale were capable of scavenging radicals, inhibiting lipid peroxidation and protecting PC12 cells against the insult by H2O2. Additionally, compound 3 could inhibit the growth of K562 and K562/ADR cells.

Animals ; Antioxidants ; toxicity ; Cell Proliferation ; drug effects ; Cytotoxins ; toxicity ; Diarylheptanoids ; isolation & purification ; metabolism ; toxicity ; Free Radicals ; metabolism ; Ginger ; chemistry ; Humans ; Hydrogen Peroxide ; metabolism ; K562 Cells ; Oils, Volatile ; pharmacology ; PC12 Cells ; Rats ; Rats, Sprague-Dawley

OBJECTIVE：To provide refere nce f or the quality control and evaluation of rice-wine processed Coptidis Rhizoma decoction piece. METHODS ：Taking 17 batches of rice-wine processed Coptidis Rhizoma decoction piece from different manufacturers as samples ，HPLC method was adopted to determine the contents of 4 kinds of alkaloids as epiberberine ，coptisine， palmatine and berberine. The compound weights of epiberberine ，berberine，palmatine and berberine were calculated by the subjective and objective combination weighting method （AHP combined with variation coefficient ）. Then the quality evaluation method was used to evaluate the quality of decoction pieces combined with the appearance of decoction pieces and the contents of 4 alkaloids. The percent mass constant was calculated and the grade of rice-wine processed Coptidis Rhizoma decoction piece was classified. RESULTS ：According to the results of content determination of 4 kinds of alkaloids ，among 17 batches of samples ，a total of 13 batches of samples met the requirements of 2015 edition of Chinese Pharmacopoeia （part Ⅳ）. Mass constants of 13 batches of qualified samples were 10.03-26.96，and the percent mass constants were 37.20％-100％. If the percent mass constant ≥ 80％ of the sample was listed as the first-class product ，the sample with the percent mass constant between 50％-＜80％ was classified as the second-class product ，and the rest was listed as the third-class product ，therefore the quality constant of first-class product was ≥21.57，that of second-class product was 13.48-＜21.57，and that of third-class product was ＜13.48. According to the grading standard ，3 batches of 13 batches of qualified samples are classified as first-class products ，6 batches are classified as second-class products ，4 batches are classified as third-class products. CONCLUSIONS ：The established subjective and objective combination weighting method and quality constant method can more scientifically and reasonably classify rice-wine processed Coptidis Rhizoma decoction piece.

Objective: To establish the in vitro porcine gastric model of submucosal eminence lesion and to evaluate its application to endoscopic submucosal dissection(ESD). Methods: Silicone rubber impression materials and steel balls with diameters of 1 cm, 2 cm, and 3 cm were used to make three pairs of spherical cavities. And then raw ground beef was put into spherical cavities and boiled for 20 minutes to make spherical mass models. Six isolated porcine stomach with esophagus and duodenum were selected. The mass models with diameters of 1 cm, 2 cm and 3 cm were imbedded respectively into the submucosa of fundus, body, and antrum of porcine stomach through the incision on serosal layer. The submucosal masses were observed by endoscopy and endoscopic ultrasonography and ESD was performed. Results: A total of 18 mass models were constructed in 6 porcine stomachs, of which 17 models were successfully established and 1 failed. Typical endoscopic characteristics of gastric submucosal eminence lesions were found in 17 models. Endoscopic ultrasonography showed that these models originated from submucosal layer and demonstrated mixed echo. There were no significant differences between mucosa of lesions and that of surrounding areas. ESD was successfully performed in the porcine gastric models of submucosal eminence lesions, and all models were not broken or detached. Conclusion The in vitro porcine gastric model of submucosal eminence lesions can well replicate disease status and provide a suitable model for study on endoscopic therapy of submucosal eminence lesion and training of endoscopists.