Reverse genetics approaches can directly manipulate the genome of virus at the gene level, making it possible to quickly, directly and thoroughly study the mechanisms of virus replication and pathogenesis. At present, many viruses of the family Reoviridae, such as mammalian orthoreovirus ( MRV) and bluetongue virus ( BTV) , have made great progress in basic viral research using the powerful tool of re-verse genetics. However, for members of the genus Rotavirus in the family Reoviridae, progress in the con-struction of reverse genetic systems has been slow. The remarkable reverse genetics system based on helper-viruses was established in 2006, and it was not until 2017 that the entirely plasmid-based reverse genetics system was successfully established. This paper briefly reviewed the development of reverse genetics systems for rotavirus and prospected the direction for future research in order to provide technical support for acceler-ating the basic research on mechanisms of rotavirus infection.

With the requirements of early diagnosis, biomarker development and functional definition, the challenge of sensitivity of immunoassay has become increasingly prominent. How to improve it to break the bottleneck has become a major challenge in the field of bioassays. Amplifying the immunosignal is the most direct method to improve detection sensitivity. Biotin-avidin system (BAS), tyramide signal amplification (TSA) and immuno-polymerase chain reaction (Im-PCR) are the most classic signal amplification techniques which significantly improved the sensitivity of immunoassays. In recent years, studies have confirmed that the sensitivity of immunoassays can be further increased by approximately three orders of magnitude with the invention of techniques including catalyzed reporter deposition-based signal amplification, nanotechnologies-based signal amplification and hybridization chain reaction-based signal amplification. Herein, we will summarize the techniques that have been developed in recent years for amplifying the signals of immunodetection and comparatively analyze their advantages and disadvantages in order to provide reference for the developed techniques transformed to clinical application and further research on ultrasensitive immunoassays.

Rotavirus (RV) is one of the leading causes of acute gastroenteritis in infants and young animals worldwide. Rotavirus infection has obvious species specificity and mainly causes diarrhea in infants and young animals. The host innate responses suppress the infection and replication of rotavirus through activating multiple signaling pathways. Meanwhile, rotavirus also antagonizes the innate immune responses in various ways. This article reviewed the mechanisms of host innate immune responses to rotavirus infection and the antagonistic mechanism of rotavirus against host innate immunity with a view to providing reference for the development of therapeutic drugs and the prevention of rotavirus infection.

Most of the human cytomegalovirus (HCMV) infection has no obvious clinical symptoms, but it can be latent for life and activated under specific conditions. HCMV active infection during pregnancy can lead to abortion, stillbirth, birth-defect and so on, which causes serious economic and social burdens. Both primary and secondary HCMV infection can lead to congenital infection of newborn, but there is still no effective method for the screening of HCMV secondary infection during pregnancy currently. Therefore, a comprehensive congenital HCMV screening for newborns is implemented for early intervention and thus reducing the consequences of congenital HCMV infection. In this paper, the methods of HCMV laboratory detection and its feasibility for neonatal screening are analyzed, in order to provide a basis for the selection of methods in neonatal congenital HCMV screening.

Rotavirus is one of the most common causes of gastroenteritis among children under five years of age worldwide. Most children are infected with rotavirus in their early childhood. Currently, no ef-fective treatment is available for diarrhea caused by rotavirus infection, and vaccination is the most effective way to prevent rotavirus infection. With the introduction of rotavirus vaccines, the morbidity and mortality of severe diarrhea in children caused by rotavirus infection are declined significantly, but these rotavirus vac-cines have significantly lower efficacy in developing countries in Africa and some countries in Asia where the mortality of rotavirus-related diseases is high than in developed countries. High titers of maternal antibodies may be one of the reasons why the efficacy of rotavirus vaccines is low in these countries. Extensive studies have been conducted on the relationship between neonatal response to rotavirus vaccine immunization and ro-tavirus-specific antibodies in the mother′s milk and serum in recent years. However, results of some studies are in conflict. In this review, we summarize the relationship between maternal antibodies and the immune responses after vaccination with rotavirus vaccines in order to provide basis for improving the efficacy of rota-virus vaccines in low-income countries.

Rotaviruses are leading causes of worldwide acute diarrhea in children younger than 5 years old, with severe consequence of social and economic burden. Vaccination is the most effective way to control rotavirus infection, however, the licensed rotavirus vaccines are ineffective in some low-income countries of Africa and Asia, where the mortality caused by rotavirus is higher than other areas. In addition, there are also safety concerns such as increased risk of intussusception. Therefore, it is urgent to improve the efficiency and safety of rotavirus vaccine to reduce the morbidity and mortality caused by rotavirus. Till now, many efforts are made to improve the effectiveness of rotavirus vaccines, and the inactive vaccine becomes the main trend in the research of rotavirus vaccine. The developments in recombinant rotavirus vaccines, especially in VP4 subunit vaccines are summarized in this review, and it could be helpful to develop effective recombinant rotavirus vaccines in further studies.

Objective : To investigate the expression and clinical significance of classic vascular endothelial growth factor A (VEGFA) and vasotropic mimicry factor matrix metalloproteinase 14 (MMP⁃14) in lung adenocarcinoma(LUAD) . Methods : The expression levels of MMP⁃14 and VEGFA genes and proteins in LUAD and their correlation with survival and prognosis were analyzed using TCGA and UALCAN databases . Serum of 69 patients with LUAD and 20 healthy subjects (control group) was collected , and the contents of MMP⁃14 and VEGFA were detected by ELISA and chemiluminescence , respectively , to analyze the correlation between the two and the clinicopathological features of tumors and their value in prediction and diagnosis of LUAD . Results: The levels of MMP⁃14 and VEGFA in LUAD tissues and serum were higher than those in control group . There were significant differences in serum VEGFA and MMP⁃14 expression levels among early stage group , advanced stage group and control group (P< 0. 001) . MMP⁃14 was higher in T3 /T4 stage than that in T1 /T2 stage (P = 0. 045) , higher in N2 /N3 stage than that in N0 /N1 stage (P = 0. 035) , and higher in the pleural metastasis group than that in the non⁃pleural metastasis group (P = 0. 034) . VEGFA level was higher in M 1 than M0 (P = 0. 025) . Elevated VEGFA level was a risk factor for LUAD (P = 0. 002) . The area under the curve (AUC) of MMP⁃14 , VEGFA and CEA alone was 0. 793 , 0. 849 and 0. 851 , respectively , and the AUC of the combined test was 0. 952 . The overall survival ( OS) and disease specific survival (DSS) of MMP⁃14 and VEGFA low expression group were longer than those of MMP⁃14 and VEGFA high expression group (P < 0. 05) . Conclusion High expression of MMP⁃14 and VEGFA in LUAD is associated with the growth , invasion and metastasis of LUAD , respectively , and has implications for survival prognosis determination .

Targeted protein degradation (TPD) technology facilitates specific and efficient degradation of disease-related proteins through hijacking the two major protein degradation systems in mammalian cells: ubiquitin-proteasome system and lysosome pathway. Compared with traditional small molecule-inhibitors, TPD-based drugs exhibit the characteristics of a broader target spectrum. Compared with techniques interfere with protein expression on the gene and mRNA level, TPD-based drugs are target-specific, efficaciously rapid, and not constrained by post-translational modification of proteins. In the past 20 years, various TPD-based technologies have been developed. Most excitingly, two TPD-based therapeutic drugs have been approved by FDA for phase Ⅰ clinical trials in 2019. Despite of the early stage characteristics and various obstructions of the TPD technology, it could serve as a powerful tool for the development of novel drugs. This review summarizes the advances of different degradation systems based on TPD technologies and their applications in disease therapy. Moreover, the advantages and challenges of various technologies were discussed systematically, with the aim to provide theoretical guidance for further application of TPD technologies in scientific research and drug development.
Animals ; Proteasome Endopeptidase Complex/metabolism* ; Protein Processing, Post-Translational ; Proteins/metabolism* ; Proteolysis ; Technology

In previous studies, we found that truncated rotavirus VP4* (aa 26-476) could be expressed in soluble form in Escherichia coli and confer high protection against rotavirus in the mouse mode. In this study, we further improved the immunogenicity of VP4* by polymerization. The purified VP4* was polymerized through incubation at 37 ℃ for 24 h, and then the homogeneity of the particles was analyzed by HPLC, TEM and AUC, while the thermal stability and antigenicity was analyzed by DSC and ELISA, respectively. Finally, the immunogenicity and protective efficacy of the polymers analyzed by a mouse maternal antibody model. The results showed that VP4* aggregated into homogeneous polymers, with high thermostability and neutralizing antibody binding activity. In addition, VP4* polymers (endotoxin <20 EU/dose) stimulated higher neutralizing antibodies and confer higher protection against rotavirus-induced diarrhoea compared with the VP4* trimers when immunized with aluminium adjuvant. In summary, the study in VP4* polymers provides a new strategy for the development of recombinant rotavirus vaccines.
Animals ; Antibodies, Viral ; Antigens, Viral ; Capsid ; Capsid Proteins ; Mice ; Polymerization ; Rotavirus ; Rotavirus Infections