Grass carp reovirus (GCRV), a disaster agent to aquatic animals, belongs to Genus Aquareovirus of family Reoviridea. Sequence analysis revealed GCRV genome segment 8 (s8) was 1296 bp nucleotides in length encoding an inner capsid protein VP6 of about 43kDa. To obtain in vitro non-fusion expression of a GCRV VP6 protein containing a molecular of fluorescence reporter, the recombinant baculovirus, which contained the GCRVs8 and eGFP (enhanced green fluorescence protein)genes, was constructed by using the Bac-to-Bac insect expression system. In this study, the whole GCRVs8 and eGFP genes, amplified by PCR, were constructed into a pFastBacDual vector under polyhedron (PH) and p10 promoters, respectively. The constructed dual recombinant plasmid (pFbDGCRVs8/eGFP) was transformed into DH10Bac cells to obtain recombinant Bacmid (AcGCRVs8/eGFP) by transposition. Finally, the recombinant bacluovirus (vAcGCRVs8/eGFP) was obtained from transfected Sf9 insect cells. The green fluorescence that was expressed by transfected Sf9 cells was initially observed 3 days post transfection, and gradually enhanced and extended around 5days culture in P1(Passage1) stock. The stable high level expression of recombinant protein was observed in P2 and subsequent passage budding virus (BV) stock. Additionally, PCR amplification from P1 and amplified P2 BV stock further confirmed the validity of the dual-recombinant baculovirus. Our results provide a foundation for expression and assembly of the GCRV structural protein in vitro.

[Objective]To evaluate the curative effect,indication and care for cage,the patients with lumbar vertebrae instability were treated by 1umbar interbody ecstatic fusion cage.[Method]There were 42 patients,male 31 and female 11,aged 3 5~78 years(averaged 46 years) and suffered with spondylolisthesis of isthmus non-union in 26 cases and degenerative spondylolisthsis in 1 6 cases,including spondylolisthsis of L3、4 5 cases,of L4、5 23 cases and of L5S1 14 cases.According to Meyerding classification,there were grade-I spondylolisthsis 29 cases and grade-Ⅱ 13 cases.All patients were treated with decompression of lumbar canal,resection of vertebral disc,insertion of ecstatic fusion cage.The curative effect was observed and evaluated.[Result]Patients were followed up for 6 months to 6 years,the results were good rate of JOA scores 95%,the recovery rate of inter-spinal height,reduction and bone fusion rate were up to 98%.[Conclusion]For lumbar spondylolisthesis of grade Ⅰ～Ⅱ,the ecstatic fusion cage has good effect on recovery of inter-spinal height,improvement of bone fusion rate,maintaining lumbar stability,relieving clinical symptom.It is a good curative method for lumbar vertebrae instability.

Objective To investigate the effect of rosiglitazone (ROS),a peroxisome poliferator- activated receptor (PPAR)?ligand,combined with all-trans retinoic acid (ATRA) on anti-angiogenesis of transplanted gastric cancer in nude mice,and to explore the mechanism of anti-angiogenesis prelimina- rily.Methods The model of xenograft tumor in nude mice were established by inoculating human gastric cancer cells line MGC803 (lower differentiated) into the back of nude mice subcutaneously.The cancer- bearing nude mice were divided randomly into 5 groups:group 1 (n=6) with no treatment;ROS treat- ment (group 2,n=6),ROS combined ATRA treatment including:low dose treatment (group 3,n= 6),moderate dose treatment (group 4,n=6) and high dose treatment (group 5,n=6).After treated for forty days,the volume change of tumor and tumor inhibition rates were observed.The expression of CD34,vascular endothelial growth factor (VEGF) in grafts were detected by immunohistochemical and calculated the difference of MVD.The mRNA expression levels of VEGF,HIF-l?were detected by RT- PCR assay accordingly.Results①The volume of tumor was significantly decreased in ROS treatment group compared with group 1 (P＜0.01).The tumor inhibition rates of group 2 were similar to group 3 (P＞0.05).With the increasing of the dose of ROS the tumor inhibition rates were increased.They were dose-dependent in specified dose-range.②ROS could inhibit angiogenesis of xenograft tumor and depress expression of mRNA of VEGF and HIF-l?.When ROS combined with ATRA,the increasing of dose of ROS,inhibiting angiogenesis of tumor and depressing expression of mRNA of VEGF and HIF-l?were found (P＜0.05).Conclusion ROS (25 mg?kg?2 d~(-1)) can inhibit the growth of tumor,and ROS combined with ATRA can further inhibit the growth of tumor,which may be through the path of PTEN by inhibiting the angiogenesis of tumor

The research and development of monoclonal antibodies (mAbs) is a rapidly developing field. From the first generation of murine mAbs to the fourth generation of fully human mAbs, the efficacy and safety of mAbs in the treatment of various diseases have been continuously improved. In order to regulate the development and evaluation of mAbs, drug regulatory agencies and pharmacopeias of America and China have tried to issue feasible test procedures and acceptance criteria for quality evaluation of mAbs and biosimilars. Mass spectrometry (MS) technique with high sensitivity, resolution, selectivity, and specificity has become an important tool to evaluate the quality characteristics of monoclonal antibody-related products or specify mAb quality. The research of MS-based monoclonal antibody study involves structure characterization, impurity analysis, pharmacokinetics/pharmacodynamics (PK/PD), etc. This review focuses on the current quality control requirements of mAb related products and the development of MS technique for mAb quality characterization and specification. It is expected to provide information and references for evaluating the quality of monoclonal antibodies under research and development.

In recent years, the biopharmaceutical industry has grown rapidly, and the market size of monoclonal antibody drugs has increased significantly. Accurate structural characterization and quality control are the supporting technologies for the development of monoclonal antibody drugs. As a significant post-translational modification of antibody drugs, glycosylation has an important influence on its efficacy, stability, and immunogenicity. The existing literature usually uses liquid chromatography-mass spectrometry to perform major glycosylation modifications of monoclonal antibody drugs. Characterization, there are few studies on low-abundance glycosylation, but the characterization and control of low-abundance glycosylation cannot be ignored. In this study, we have established a qualitative and quantitative analysis technology for N-glycans based on RapiFluor-MS reagent-labeled monoclonal antibody drugs. This method has a short sample processing time and high sensitivity. It can not only characterize the main glycoforms of three monoclonal antibody drugs (adalimumab, bevacizumab, and trastuzumab) but also can quantify low-abundance N-glycans. The results of the study showed that the main glycoforms specified in the Pharmacopoeia could be detected in different batches of monoclonal antibody drugs, but the content of N-glycans in different batches of samples is not identical. After that, we analyzed the N-glycans connection sites and glycoforms at the intact glycopeptide level, further enriching the N-glycans structure information of the monoclonal antibody. The qualitative and quantitative analysis technology of N-glycans based on RapiFluor-MS reagent-labeled monoclonal antibody drugs can realize the in-depth characterization and control of glycosylation modification of monoclonal antibody drugs.

Animals ; Antioxidants ; pharmacology ; Female ; Hypoxia ; physiopathology ; Male ; Mice ; Panax ; chemistry ; Random Allocation ; Reperfusion Injury ; prevention & control ; Saponins ; isolation & purification ; pharmacology

Animals ; Antioxidants ; pharmacology ; Brain ; metabolism ; Brain Ischemia ; metabolism ; Male ; Malondialdehyde ; metabolism ; Panax ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; Saponins ; pharmacology ; Superoxide Dismutase ; metabolism

Adult ; Aged ; Breast Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Female ; Gene Amplification ; Genital Neoplasms, Male ; genetics ; metabolism ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Paget Disease, Extramammary ; genetics ; metabolism ; pathology ; surgery ; Paget's Disease, Mammary ; genetics ; metabolism ; pathology ; surgery ; Penile Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Receptor, ErbB-2 ; genetics ; metabolism ; Scrotum ; Vulvar Neoplasms ; genetics ; metabolism ; pathology ; surgery

Animals ; Brain ; metabolism ; Brain Ischemia ; metabolism ; Excitatory Amino Acids ; metabolism ; Female ; Male ; Panax ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Reperfusion Injury ; prevention & control ; Saponins ; isolation & purification ; pharmacology

Objective To understand the relation between smoking and serum uric acid level and to investigate whether the ser-um uric acid has the correlation with the gender,age and smoking history.Methods The data of the gender,age,blood uric acid in1 847 individuals aged 20-80 years with the healthy physical examination and without underlying diseases were performed the statisti-cal analysis.Results With male and female as the research objects,the serum uric acid level of smokers were higher than that of non-smokers and occasional smokers,the difference was statistically significant;the serum uric acid level had no statistically signifi-cant difference between smokers and occasional smokers;the serum uric acid level had no statistically significant difference among non-smoking,occasional smoking and smoking groups for males as the research objects alone;to divide the male subjects into groups according to age,the serum uric acid level of non-smokers,occasional smokers and smokers were not statistically significant among all age groups;serum uric acid level showed the increasing trend with the increase of smoking history,but there was no statistically significant difference.Conclusion The serum uric acid level of smokers is significantly higher than that of non-smokers and occa-sional smokers with male and female as the research objects;the difference in serum uric acid level between smokers and occasional smokers has no statistical significance;excluding the gender factor interference,the serum uric acid level of males is not affected by smoking or age;serum uric acid mean value demonstrates the increasing trend with the increase of smoking history.