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Objective:To screen and identify H-2 d-restricted T cell epitopes in fusion (F) and attachment (G) glycoproteins of Nipah virus (NiV) in mice. Methods:The complete peptides (single peptide contains 15 amino acids, and 10 amino acids were repeated in the front and back peptides) derived from F and G antigens were mixed into peptide libraries. BALB/c mice were immunized with DNA vaccines expressing NiV F and G proteins alone and in combination. The full sequence peptide libraries of F and G antigens were mixed into peptide pools by matrix design, and spleen cells of immunized mice were collected and analyzed by IFN-γ ELISPOT assay to detect the dominant H-2 d-restricted epitope peptides. Results:Twelve dominant H-2 d-restricted peptides were screened from the F protein-specific peptide library and the 56th peptide produced the strongest reaction. Four dominant peptides were screened from the G protein-specific peptide library and the 72nd peptide produced the strongest reaction. Conclusions:In this study, 12 F antigen-specific and 4 G antigen-specific H-2 d restricted dominant T cell epitopes of NiV were screened and identified by IFN-γ ELISPOT, which could provide reference for immunological analysis of NiV and vaccine research.
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Objective:To explore ultrasonographic diagnostic characteristics of ovarian epithelial tumors and establish prediction models.Methods:The ultrasonographic images of 427 cases from multicenter with ovarian epithelial tumors confirmed by pathology from January 2015 to July 2019 were retrospectively analyzed according to the International Ovarian Tumor Analysis (IOTA). Ultrasonographic signs with distinguishing significance were obtained through univariate analysis and included into multivariate Logistic regression analysis to obtain important ultrasonagraphic indicators for distinguishing borderline, benign and malignant ovarian tumors, and to establish prediction models.Results:The microcystic pattern of papillary projections and solid components was the diagnostic characteristic between borderline and benign, malignant ovarian epithelial tumors( OR value 10.97 and 19.22, respectively). Irregular morphology, septa thickness, solid lesions, rich blood supply and ascites were diagnostic characteristics between benign and malignant tumors, with the irregular morphology having the highest value. Irregular morphology, large papillary, septa thickness and rich blood supply could be used to identify borderline and malignant tumors. At the same time, irregular morphology was the valuable sign to distinguish borderline and benign tumors. In this study, the total coincidence rate of the proposed model was 72.4%, among which the predicted coincidence rate of the borderline model was 57.2%, 78.6% for benign, and 80.7% for malignant. Conclusions:The microcystic pattern of papillary projections and solid components are the specific sonographic characteristics of borderline ovarian tumors. Irregularity, solid lesions, rich blood supply and ascites have important value in differentiating ovarian epithelial tumors. The prediction models of benign, malignant and borderline ovarian tumors in this study have higher diagnostic efficacy.
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Influenza poses a serious threat to global public health and causes serious economic los-ses. Vaccination is the most effective measure to prevent influenza. However, the mismatch between the vac-cine strain and the epidemic strain, which results from antigenic drift and antigen conversion of influenza vi-rus, often invalidates the conventional vaccine stockpiles. mRNA vaccine is a relatively safe platform com-posed of nucleic acid. Improvements in genetic engineering technology and novel delivery systems have great-ly promoted the research and development of mRNA vaccines against influenza. mRNA vaccines are promis-ing candidates for the rapid and efficient prevention of seasonal influenza epidemics and influenza pandemics.
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Aim To investigate the effects of cajanonic acid A (CAA) on lipid metabolism in murine 3T3-L1 adipocytes. Methods 3T3-L1 cells induced to differ-entiated into mature adipocytes were treated with CAA in different dosages for 48 h, then total lipids as well as triglyceride, free fatty acid and glycerol were meas-ured. The expression levels of genes related to lipid metabolism were quantitatively analyzed by real-time fluorescent quantitative polymearase chain reaction ( RTFQ-PCR) . Results Total lipids and triglyceride in 3T3-L1 adipocytes were markedly reduced by CAA. The release of free fatty acid and glycerol was lower than that of control. This coincided with decreased mRNA levels of the key enzymes involved in de novo lipogenesis ( acetyl CoA carboxylase and fatty acid syn-thase) , fatty acid uptake ( lipoprotein lipase) , and li-polysis ( hormone sensitive lipase and adipose triglycer-ide lipase ) . While the expression of fatty acid oxida-tive genes including acyl CoA oxidase and carnitine palmitoyl transferase1 was increased after CAA treat-ment. Conclusion CAA may inhibit lipogenesis and lipolysis,reduce circulating free fatty acid and improve the lipid metabolism in adipocytes by regulating gene expressions.
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Objective To evaluate the application value of three‐dimensional ultrasonography OmniView technology in diagnosis of fetal cleft lip/palate . Methods Three‐dimensional volume data was acquired from 100 normal singleton fetuses and 18 cleft lip/palate fetuses ,and was analysed by OmniView technology . Two‐dimensional ultrasonography and three‐dimensional OmniView technology were compared in the displaying rate of different planes of the lip and palate and the diagnosis accordance rate of cleft lip/palate.Results ①ThedisplayingrateofthelipandpalateofOmniViewtechnologywashigherthanthatof two‐dimensional ultrasonography ( the displaying rates were 64% -88% and 30% -65% ,respectively , P 0 .05) . Conclusions Three‐dimentional ultrasonography OmniView technology can improve the displaying rate of the hard and soft palate .The application of this technology in the fetuses who has a high risk of cleft palate may be beneficial to the diagnosis of cleft palate .
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BACKGROUND:Guidances for design of Chinese people’s temporomandibular joint prosthesis are urgently needed because foreign prosthesis is unsuitable for Chinese people due to variant anatomy parameters. OBJECTIVE:To provide a reference for improving the design of artificial temporomandibular joint through computer three-dimensional digital measures of the anatomy parameters of the temporomandibular joint. METHODS:Continuous CT data with layer thick 0.625 mm of 30 normal temporomandibular joints were selected. Three-dimensional digital reconstruction and eight related anatomy parameter measurements of the articular fossaandcondylar of the joints were performed using Mimics10.01 software. RESULTS AND CONCLUSION:The internal and external diameter of the mandibular fossa, head of mandible and neck of mandible were greater thantheanterior and posteriordiameter. The mandibular foramen was located at middle-posterior of the ramus ofthe mandible. The distance between head of the mandible and coronoid process was longer than the depth of the mandibular notch. There were no significant differences between two sidesregardingal above-mentioned parameters. There are certain regular patterns in anatomy parameters of the temporomandibular joint. The study results wil provide a reference for improving the design of artificial temporomandibular joint.
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To character a novel chimera(1b/2a) hepatitis C virus cell culture (HCVcc) system carrying envelope E1E2 coding gene from Hebei strain of China, chimera HCVcc (cHCVcc) was developed from Huh7.5-CD81 cells after transfection with in vitro transcribed full-length 1b/2a chimera RNA, which carrying envelope E1E2 coding gene from Hebei strain of China. Then the replication, expression and infectious titer of serial passage HCVcc were assessed by Real Time RT-PCR, indirect immunofluorescence assay (IFA) and Western blotting (WB). In addition, chimeric envelope gene from HCVcc was sequenced after serial passage. We found that the number of HCV positive focus increased gradually in cell post-transfection with chimera HCVcc (1b/2a) RNA and reach a peak platform (80% to 90%) at 41 days post-transfection; the expression of HCV protein was also confirmed by WAB during serial passage. At meantime, HCV RNA copy number in the supernatant peaked at 10(4)-10(7) copies/mL and the highest infectious titer of this 1b/2a cHCVcc reinfection were tested as 10(4) ffu/mL. Sequence analysis indicated 6 of adaptive amino acid substitutes occur among chimeric envelope E1E2 during serial passages. We con:luded that a novel 1b/2a chimera HCVcc carrying envelope E1E2 coding gene from Hebei strain of China was developed and its infectious titer increased after serial passage of HCVcc. This novel cHCVcc will be an effective tool for further evaluation of anti-virus drugs and immune effects against the major genotype from Chinese.
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Humans , Cell Line , China , Hepacivirus , Genetics , Metabolism , Hepatitis C , Virology , Serial Passage , Viral Envelope Proteins , Genetics , MetabolismABSTRACT
Objective To express the N protein of middle east respiratory syndrome coronavirus ( MERS-CoV) in prokaryotic expression system and evaluate the immunogenicity of the purified recombinant N protein.Methods The gene encoding N protein of MERS-CoV was synthesized and cloned into the vector pET30a to construct the recombinant expression plasmid pET30a-MERS-CoV-N.The transformed E.coli BL21 ( DE3) strains carrying expression plasmid were induced by IPTG to express N protein.The expressed protein was purified by using affinity chromatography.SDS-PAGE and Western blot assays were used to iden-tify the expressed N protein and evaluate its immunogenicity.Results The recombinant N protein was suc-cessfully expressed in soluble form with the size of 46×103 .The results of Western blot assay showed that the recombinant N protein could specifically react with rabbit serum samples positive for antibodies against N protein B-cell epitope peptide and mouse anti-His tag antibodies.No positive band against N protein was found when primary antibody was used with thirty adult serum samples from Beijing in 2008.Conclusion N protein of MERS-CoV was successfully expressed in prokaryotic expression system.The successful expres-sion of N protein laid the foundation for immunological diagnosis of N protein of MERS-CoV and researches on its immunogenicity.
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<p><b>OBJECTIVE</b>To establish quality control criteria for medicinal herb Cajanus cajan based on the determination of longistylin A and longistylin C, two bioactive and specific stilbenes of the plant.</p><p><b>METHOD</b>Longistylin A and longistylin C were obtained from the leaves of C. cajan by silica gel column chromatography and identified as marker compounds of this plant by spectroscopic analysis. A RP-HPLC method was established to determine the two compounds.</p><p><b>RESULT</b>Longistylin A and longistylin C were well separated on a Thermo BDS Hypersil C18 column (4.6 mm x 250 mm, 5 microm) with a mobile phase methanol-water (8:2), and showed good linearity in the range of 0.00288 - 0.0576 microg and 0.0112 - 0.224 microg, respectively. The average recoveries were 98.9% and 97.2% with RSD of 2.4% and 2.2% for these two compounds, respectively.</p><p><b>CONCLUSION</b>The established analysis method is simple and accurate, whicn can be used for quality control of C. cajan.</p>
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Cajanus , Chemistry , Chromatography, High Pressure Liquid , Methods , Diethylstilbestrol , Drugs, Chinese Herbal , Plant Leaves , Chemistry , Plants, Medicinal , ChemistryABSTRACT
The physicochemical environment and action are similar between the traditional decoction and the extract technics with water or alcohol in the production of Chinese patent drug. Different heating time inevitably differs Chinese patent drug from its decoction; and the alteration of extracting dissolvent make great changes in the chemical constitution. All these lead to the change in the nature of a Chinese patent drug. The authors hold that it is difficult to embody exactly the aim of the prescription of Chinese drug in the existing production technology of Chinese patent drug. It is necessary to advance innovative thoughts of adopting modern technology to extract effective ingredients from single Chinese drug and in the reference of traditional decoction, recombining the composition and dosage of Chinese patent drug.