Identification of a novel Getah virus by Virus-Discovery-cDNA random amplified polymorphic DNA (RAPD).

BACKGROUND: The identification of new virus strains is important for the study of infectious disease, but current (or existing) molecular biology methods are limited since the target sequence must be known to design genome-specific PCR primers. Thus, we developed a new method for the discovery of unknown viruses based on the cDNA--random amplified polymorphic DNA (cDNA-RAPD) technique. Getah virus, belonging to the family Togaviridae in the genus Alphavirus, is a mosquito-borne enveloped RNA virus that was identified using the Virus-Discovery-cDNA RAPD (VIDISCR) method. RESULTS: A novel Getah virus was identified by VIDISCR from suckling mice exposed to mosquitoes (Aedes albopictus) collected in Yunnan Province, China. The non-structural protein gene, nsP3, the structural protein gene, the capsid protein gene, and the 3'-untranslated region (UTR) of the novel Getah virus isolate were cloned and sequenced. Nucleotide sequence identities of each gene were determined to be 97.1-99.3%, 94.9-99.4%, and 93.6-99.9%, respectively, when compared with the genomes of 10 other representative strains of Getah virus. CONCLUSIONS: The VIDISCR method was able to identify known virus isolates and a novel isolate of Getah virus from infected mice. Phylogenetic analysis indicated that the YN08 isolate was more closely related to the Hebei HB0234 strain than the YN0540 strain, and more genetically distinct from the MM2021 Malaysia primitive strain.

Acute respiratory distress syndrome after liver transplantation: etiology, prevention and management.

OBJECTIVE: To study the etiology, prevention and management of acute respiratory distress syndrome (ARDS) after liver transplantation. METHODS: The clinical data of 104 patients with end-stage liver diseases who had had liver transplantations were retrospectively reviewed. RESULTS: Seventeen patients (16.3%, 17/104) altogether were diagnosed as having ARDS after liver transplantation. Ten of them developed ARDS within 24 hours, of whom 1 died during the operation, and 7 developed ARDS 3 or 4 days after they were extubated and when methylprednisolone was tapered. Fourteen of the 17 ARDS patients (14/17) were found to have overloaded crystalloid infusion, massive transfusion of blood or blood products such as plasma, platelets, in addition to a prolonged surgical time secondary to serious bleeding during the diseased liver removal without evidence of active infection. One was found to have serious systemic infection and operatively disseminated intravascular coagulation. Four of the recipients developed ARDS suddenly when intravenous cyclosporine was given on the 3rd day after operation. One patient of the 4 had all of the aforementioned conditions. Two patients suffered from gastric aspiration. Five (30%, 5/17) of them survived ARDS with the combined treatment consisting of positive end-expiratory pressure mechanical ventilation suctioning as much edema fluid or sputum as possible, administration of diuretics, bolus of corticosteroids, and culture-based antibiotics. Hemeodialysis was indicated for patients with oliguric renal failure. CONCLUSIONS: ARDS is a serious multifactoral complication after liver transplantation with a high mortality and fatality. The most likely cause is fluid overload from crystalloid liquid infusion or massive transfusion. The other predisposing or contributing factors include sepsis, IV use of cyclosporine, fast tapering of corticosteroids, and gastric aspiration. Other factors such as transfusion-related acute lung injury (TRALI), and reperfusion syndrome of the newly implanted liver may also contribute. Though the treatment should primarily be supportive in nature, it is helpful to understand the predisposing and contributing factors and to aid in prevention, management and treatment.

[Comparative study of three kinds of outflow tract reconstruction of hepatic veins in piggyback liver transplantation].

OBJECTIVE: To study the effects of 3 different operational patterns of piggyback liver transplantation (PBLT) used to reconstruct backflow of hepatic veins. METHODS: Sixty-three operations of PBLT were performed on 59 patients with terminal hepatic diseases after three operational patterns: EEAT [the suprahepatic inferior vena cava (sup-H-IVC) of donor is anstomosed with the plasticized hepatic vein of recipient end-to-end, also called standard PBLT, SPBLT] in 17 cases, ESAT (the sup-H-IVC of donor is anastomosed with the sup-H-IVC of recipient end-to-side) in 12 cases, and SSAT [the retrohepatic IVC (RHIVC) of donor is anastomosed with the RHIVC of recipient side-to-side] in 32 cases, the latter two patterns being called ameliorative PBLT (APBLT) jointly. The effects were analyzed. RESULTS: Complications, such as backflow obstruction of hepatic vein and delayed recovery of liver function, were observed in the EEAT and ESAT groups, but not in the SSAT group. CONCLUSION: The SSAT pattern of PBLT is easy to perform and advantageous to avoid the technical maladies of the other 2 patterns and postoperative complications, and provides assurance of recovery after operation.

Inhibition of mCD14 inhibits TNFα secretion and NO production in RAW264.7 cells stimulated by Brucella melitensis infection.

Brucellosis, caused by Brucella spp., is an important disease affecting not only human health, but also a number of animal species around the world. A receptor for LPS of Brucella and important innate immune molecule, CD14, has been implicated in the initiation of the inflammatory response to sepsis. Evidence indicates that upstream inhibition of the LPS initiated inflammatory pathway is an effective therapeutic approach for attenuating damaging immune activation. The aim of this study was to explore the possibility of using RNA interference (RNAi) targeting mCD14 as a strategy for inhibiting the secretion of tumor necrosis factor alpha (TNFα) and the production of nitric oxide (NO) from Brucella-stimulated RAW264.7 cells and attenuating damaging immune activation. The current study stably incorporated mCD14-shRNA-224 into the RAW264.7 cell line by lentiviral gene transfer to successfully knockdown mCD14, and was then challenged with Brucella melitensis M5-90. The secretion of TNFα, interleukin (IL)-12, CXCL1/KC, and inducible nitric oxide synthase (iNOS) protein expression, and NO production were evaluated. The mCD14-shRNA-224 knockdown was shown to effectively inhibit B. melitensis M5-90-stimulated TNFα release, iNOS protein expression, and NO production, but no significant differences were observed for IL-12 and CXCL1/KC. These findings provide the basis for the development of RNAi-based prophylaxis and therapy for B. melitensis induced inflammatory disease.