Sequences analysis revealed Grass carp reovirus (GCRV) s10 was 909 nucleotides coding a 34 kDa protein denoted as VP7, which was determined to be a viral outer capsid protein (OCP). To obtain expressed OCP in vitro, a full length VP7 gene was produced by RT-PCR amplification, and the amplified fragment was cloned into T7 promoted prokaryotic expression vector pRSET. The recombinant plasmid,which was named as pR/GCRV-VP7,was then transformed into E.coli BL21 host cells. The data indicated that the expressed recombinant was in frame with the N-terminal fusion peptide. The over-expressed fusion protein was produced by inducing with IPTG, and its molecular weight was about 37kDa, which was consistent with its predicted size. In addition, the fusion protein was produced in the form of the inclusion body with their yield remaining steady at more than 60% of total bacterial protein. Moreover,the expressed protein was able to bind immunologically to anti-his-tag monoclonal antibody (mouse) and anti-GCRV serum (rabbit). This work provides a research basis for further structure and function studies of GCRV during entry into cells.

Grass carp reovirus (GCRV), a double stranded RNA virus that infects aquatic animals, often with disastrous effects, belongs to the genus Aquareovirus and family Reoviridea. Similar to other reoviruses, genome replication of GCRV in infected cells occurs in cytoplasmic inclusion bodies, also called viral factories. Sequences analysis revealed the nonstructural protein NS80, encoded by GCRV segment 4, has a high similarity with uNS in MRV(Mammalian orthoreoviruses), which may be associated with viral factory formation. To understand the function of the uNS80 protein in virus replication, the initial expression and identification of the immunogenicity of the GCRV NS80 protein inclusion forming-related region (335.742) was investigated in this study. It is shown that the over-expressed fusion protein was produced by inducing with IPTG at 28oC. In addition, serum specific rabbit antibody was obtained by using super purified recombinant NS80(335.742) protein as antigen. Moreover, the expressed protein was able to bind to anti-his-tag monoclonal antibody (mouse) and NS80(335-742) specific rabbit antibody. Further western blot analysis indicates that the antiserum could detect NS80 or NS80C protein expression in GCRV infected cells. This data provides a foundation for further investigation of the role of NS80 in viral inclusion formation and virion assembly.

Objective To observe the efficacy, benefits and shortcomings of pressurized air therapy for neo- natal hypoxic ischemic encephalopathy (HIE). Methods One hundred and nine neonates with HIE were treated with hyperbaric oxygen (HBO) with a continuing oxygen supplement from an improved oxygen mask plus a 2.5-1itre breathing sacculus proprius in a large air-pressurized oxygen chamber. Among them there were 70 cases treated with 3-6 courses of HBO + drug therapy, 39 cases treated with a single session of HBO + drug therapy, and 32 treated with drug therapy alone. Motor development was assessed using the Chinese infantile intelligence development test scale at the ages of 3, 6 and 12 months. Results The sequential HBO + drug group achieved significantly better average motor development than the single session group or the drug only group. There was mo significant difference between the single treatment group and the drug only group. The proportion of abnormal CT results 12 months after treatment was significantly higher in the drug only group than in the sequential HBO + drug group. Conclusion Sequential HBO + drugs therapy with the improved oxygen mask is preferable to a single session of HBO + drug treatment or drug therapy alone.

Objective To assess the prognostic benefits of intraoperative radiotherapy (IORT) with electron beam among patients with unresectable locally advanced pancreatic cancer.Methods Between January 2009 and December 2014,167 patients with unresectable locally advanced pancreatic cancer received IORT with electron beam (10-20 Gy) in our hospital.After surgery,12 patients were treated with external beam radiotherapy,56 patients with chemoradiotherapy (CRT),and 17 patients with chemotherapy.Overall survival (OS),local recurrence,and toxicities were retrospectively analyzed.The Kaplan-Meier method was used to calculate survival rates,the log-rank test was used for survival difference analysis and univariate prognostic analysis,and the Cox model was used for multivariate prognostic analysis.Results The follow-up rate was 100％.The median OS time was 10.3 months,and the 2-year OS rate was 22％.The median progression-fiee survival (PFS) time was 6.3 months,and the 2-year PFS rate was 9.9％.The cancer-specific survival (CSS) time was 11.2 months,and the 2-year CSS rate was 23.6％.In the patients treated with IORT alone at doses of＜15 Gy,15 Gy and＞15 Gy,the median OS times were 6.2 months vs.9.1 months vs.22.2 months,and the 1-year OS rates were 10.0％ vs.39.6％ vs.74.4％ (P=0.000).Among the patients receiving postoperative adjuvant therapy,those treated with IORT+CRT had the best survival,with a median OS time of 11.6 months (P=0.033).The univariate analysis showed that IORT dose (P =0.000),tumor size (P =0.006),and IORT applicator diameter (P =0.007) were prognostic factors.The multivariate analysis showed that IORT dose (P=0.000) and IORT combined with CRT (P=0.006) were independent prognostic factors.Conclusions IORT with electron beam is an effective and safe treatment strategy for unresectable locally advanced pancreatic cancer.After protecting surrounding organs,increasing the IORT dose can improve the survival.IORT combined with CRT should be recommended because it improves survival for unresectable locally advanced pancreatic cancer without increasing toxicities.

Objective:To investigate the clinical value of integrated traditional Chinese and Western medicine based on the concept of enhanced recovery after surgery (ERAS) in the perioperative period of total knee arthroplasty.Methods:Sixty patients who underwent primary unilateral total knee arthroplasty in Tonglu Hospital of Traditional Chinese Medicine from July 2018 to June 2019 were included in this study. They were randomized as odd numbers to the control group ( n = 30) and even numbers to the observation group ( n = 30). The control group was treated with conventional rehabilitation treatment, while the observation group was subjected to integrated traditional Chinese and Western medicine based on the concept of ERAS in the perioperative period of total knee arthroplasty. The perioperative pain, related complications, hospitalization expenses and length of hospital stay were recorded in each group. The Visual Analogue Scale (VAS) was used to evaluate the degree of pain and range of motion of the knee joint. The Hospital for Special Surgery (HSS) knee score was also measured to evaluate the curative effects. Results:At 3 and 7 days, 2 and 6 weeks after surgery, knee range of motion score was (63.40 ± 2.80) points, (86.20 ± 4.40) points, (90.30 ± 6.48) points, (100.5 ± 3.39) points, respectively, and at 6 weeks after surgery, HSS knee score was (89.40 ± 5.18) points, in the observation group, which were significantly higher those in the control group [(48.50 ± 4.20) points, (55.40 ± 3.58) points, (77.50 ± 4.38) points, (87.60 ± 4.58) points, (70.50 ± 6.44) points, t = 0.029, 0.013, 0.032, 0.039, 0.042, all P < 0.05]. In the observation group, the VAS score in resting state at 6, 12, 24 and 48 hours after surgery were (3.62 ± 0.40) points, (2.41± 0.52) points, (2.05 ± 0.62) points, (1.93 ± 0.28) points respectively, and the VAS score in active state at 24 and 48 hours were (2.15 ± 0.21) points and (1.71 ± 0.39) points, respectively, which were significantly lower than those in the control group [(5.71 ± 0.63) points, (4.60 ± 0.31) points, (3.84 ± 0.22) points, (3.30 ± 0.21) points, (5.50 ± 0.49) points, (4.80 ± 0.21) points, t = 0.040, 0.035, 0.046, 0.042, 0.027, 0.024, all P < 0.05]. The proportions of patients experiencing postoperative urinary retention, mental disorder, or blood transfusion, length of hospital stay, and hospitalization expenses in the observation group were lower or shorter than those in the control group ( χ2 = 0.003, 0.005, 0.017, t = 0.040, 0.048, all P < 0.05). Conclusion:Integrated traditional Chinese and Western medicine based on the concept of ERAS can mitigate postoperative pain, reduce perioperative complications, and promote the recovery of knee joint function, and thereby deserves clinical application.

BACKGROUND: Radiation enteritis (RE) is a common complication of abdominal or pelvic radiotherapy, which when severe, could be life-threatening. Currently, there are no effective treatments. Studies have shown that mesenchymal stem cells (MSCs)-derived exosomes (MSC-exos) exhibit promising therapeutic effects in inflammatory diseases. However, the specific role of MSC-exos in RE and the regulatory mechanisms remain elusive. METHODS: In vivo assay was carried out by injecting MSC-exos into the total abdominal irradiation (TAI)-induced RE mouse model. For in vitro assay, Lgr5-positive intestinal epithelial stem cells (Lgr5+ IESC) were extracted from mice, followed by irradiation along with MSC-exos treatment. HE staining was performed to measure histopathological changes. mRNA expression of inflammatory factors TNF-a and IL-6 and stem cell markers LGR5, and OCT4 were quantified by RT-qPCR. EdU and TUNEL staining was performed to estimate cell proliferation and apoptosis. MiR-195 expression in TAI mice and radiation-induced Lgr5+ IESC was tested. RESULTS: We found that the injection of MSC-exos inhibited inflammatory reaction, increased stem cell marker expression, and maintained intestinal epithelial integrity in TAI mice. Furthermore, MSC-exos treatment increased the proliferation and simultaneously suppressed apoptosis in radiation-stimulated Lgr5+ IESC. MiR-195 expression increased by radiation exposure was decreased by MSC-exos therapy. MiR-195 overexpression facilitated the progress of RE by counteracting the effect of MSC-exos. Mechanistically, the Akt and Wnt/b-catenin pathways inhibited by MSC-exos were activated by miR-195 upregulation. CONCLUSION MSC-Exos are effective in treating RE and are essential for the proliferation and differentiation of Lgr5+ IESCs. Moreover, MSC-exos mediates its function by regulating miR-195 Akt b-catenin pathways.