ABSTRACT
Bombyx mori bidensovirus (BmBDV) has been identified as causing chronic densonucleosis in Bombyx mori specifically. The replication mechanism of BmBDV remains unknown. Its genome comprises two single stands DNA (VD1 and VD2). In order to rescue infectious virions in vitro, we obtained the total viral DNA extracted from the BmBDV-infected larvae midguts, subsequently cloned the full-length sequence of BmBDV genome fragments by PCR and constructed recombinant plasmids pMD18T-VD1 and pUC-VD2. The linear genome fragments were obtained by digesting recombinant plasmids with corresponding restriction enzymes, and then collectively transfected BmN cells by the method of liposome-embedding. We determined the replication of the virus gene by PCR with the template of demethylated total DNA extracted from the post-transfect BmN cells. Meanwhile, we collected the total proteins from the post-transfect BmN cells and the larvae midgut of feeding the post-transfect BmN cells to perform Western blotting analysis, and detected the expression of viral genes. Here we firstly confirm that infectious virions can be rescued in BmN cells by linear co-transfect method.
Subject(s)
Animals , Bombyx , DNA, Viral , Densovirus , Larva , Transfection , Virion , Virus CultivationABSTRACT
Objective To investigate the outcome of damage control sugery in patients with acute superior mesenteric occlusion (ASMO).Methods Seventeen patients with acute superior mesenteric occlusion from June 2008 to May 2012 were reviewed retrospectively.Diagnosis was confirmed by CT and/or DSA.Patients were divided into two groups:seven patients in damage control group (shortening surgical time,bowel exteriorization,temporary abdominal closure) ; ten patients in conventional surgical treatment group.The treatment outcomes of the two groups were compared.The survival of all patients was followed up for 6 months to 5 years at outpatient clinic.Results The overall mortality rate was (5/17) and surgical complications rate was (8/17).Operation time was (97 ±42) min and (236 ± 137) min(t =-4.72,P ＜0.01,P ＜ 0.05) respectively in damage control group and conventional surgical treatment group.Intestine resection length was (114 ± 94) cm and (229 ± 93) cm (t =-2.49,P =0.03,P ＜ 0.05) respectively.Complications rate was (1/7) and (7/10) (Fisher's dxact test P =0.05,P ≤0.05) respectively; mortality was (1/7) and (4/10) (Fisher's dxact test P =0.34,P ＞ 0.05) respectively.Conclusions Although damage control does not significantly reduce mortality,but the rational use of damage control can effectively reduce operation time,shorten intestine resection length and decrease surgical complications.