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.

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.

Objective:To investigate the application value of "four doors of the liver" approach in the laparoscopic anatomical hepatectomy.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 52 patients with liver cancer who were admitted to West China Hospital of Sichuan University from September 2018 to September 2019 were collected.Patients underwent laparoscopic anatomical hepatectomy by opening "four doors of the liver" approach. There were 36 males and 16 females, aged (53±16)years, with a range from 35 to 78 years. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up and survival. Follow-up using outpatient examination or telephone interview was conducted to detect the physical situations, liver function and recurrence of liver cancer in patients up to March 2020. Measurement data with normal distribution were represented as Mean± SD, and measurment data with skewed distribution were represented as M (range). Count data were expressed as absolute numbers or percentages. Results:(1) Surgical situations: all the 52 patients underwent laparoscopic anatomical hepatectomy successfully, without perioperative death. Eight and 8 patients underwent laparoscopic left hemihepatectomy and right hemihepatectomy by opening "the first door of the liver" respectively, the operation time of which was (151±31)minutes and (190±43)minutes, the volume of blood loss was (151±20)mL and (361±51)mL. Eight and 8 patients underwent laparoscopic left inner hepatic lobotomy and left outer hepatic lobotomy by opening "the second door of the liver" respectively, the operation time of which was (171±41)minutes and (90±26)minutes, the volume of blood loss was (221±31)mL and (111±21)mL. Eight and 8 patients underwent laparoscopic right posterior hepatic lobotomy and right anterior hepatic lobotomy by opening "the third door of the liver" respectively, the operation time of which was (172±29)minutes and (220±40)minutes, the volume of blood loss was volume of (351±41)mL and (451±47)mL. Four patients underwent laparoscopic hepatic caudate lobotomy by opening "the fourth door of the liver" , the operation time of which was (246±36)minutes, the volume of blood loss was (261±31)mL. None of the 52 patients had blood transfusion. (2) Postoperative situations: all the 52 patients recovered well after surgery, with no complications such as bleeding, biliary fistula, infection or liver failure. The duration of postoperative hospital stay was (7±4)days. (3) Follow-up and survival: all the 52 patients were followed up for 6-17 months, with a median follow-up time of 10 months. At 6 months after operation, all the 52 patients achieved of Eastern Cooperative Oncology Group performance status grade 1, Child-Pugh A of liver function, without tumor recurrence or metastasis. The overall survival rate was 100%(52/52).Conclusion:It is safe and feasible to perform laparoscopic anatomical hepatectomy by the "four doors of the liver" approach.

We report a case of a newborn baby who suffered from hemolytic disease of fetus and newborn (HDFN) caused by anti-Di a. The baby presented with worsening jaundice started at three hours after birth and was transferred to Dongguan Maternal and Child Health Care Hospital. The newborn's hemoglobin (Hb) was 82 and 76 g/L at five and nine hours after birth, and the total bilirubin (TBIL) was 243.2 and 309.8 μmol/L, respectively. Blood samples of the newborn and the parents were collected for HDFN immunohematology test twelve hours after birth. They showed that the newborn and the father's blood type was A and RhDCCee, while the mother was A and RhDCcee. Direct antiglobulin test (DAT) indicateda strong positive for the newborn and negative for the parents. The reaction of the reagent to red blood cells for antibody screening with the patient's plasma, red cells eluate, and the mother's plasma were all negative, but were positive with the father's red blood cells. The newborn was recovered after treating with phototherapy, intravenous immunoglobulins and urgent blood exchange (the exchanged blood was the same ABO and RhD blood type and cross-matched). The newborn's plasma and red cells eluate were collected before blood exchange, and the mother's plasma were used to assess the red blood cells reaction, and IgG anti-Di a was identified in each sample. Di a blood typing was positive for the newborn and the father, and negative for the mother. Therefore, the newborn was diagnosed as HDFN caused by anti-Di a.

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