To obtain human antibodies against the Gn protein of Severe fever with thrombocytopenia syndrome virus (SFTSV) with phage display technology, this study aimed to screen anti-Gn protein antibodies from an anti-SFTSV Fab human phage display library. Antibody genes were identified by sequence analysis and the specificity of antibodies was confirmed by ELISA. The Fab antibody genes were cloned into the HL51-14 vector and expressed in a mammalian cell expression system. IgG antibodies were then purified by protein A affinity chromatography,and the results were further confirmed by ELISA,IFA,western blotting assays and micro-neutralization tests. The results showed that, after three rounds of panning, there were 390 human Fab antibodies against SFTSV particles, of which 364 were specific for nucleoprotein. Coated with the Gn protein, eight different Fab antibodies specific for Gn protein were obtained after the determination of the subtype and subclass of antibodies by gene sequencing; five of these antibodies were from the Lambda library and three were from the Kappa library. The eight IgG antibodies could specifically bind to Gn protein according to the ELISA, IFA and Western blotting assays. The micro-neutralization test showed that these eight antibodies had no neutralizing activity,but they could still provide a reference for research in human monoclonal antibodies against SFTSV.
Antibodies ; genetics ; immunology ; Bunyaviridae Infections ; genetics ; immunology ; virology ; Cell Line ; Cloning, Molecular ; Humans ; Immunoglobulin Fab Fragments ; genetics ; immunology ; Immunoglobulin G ; genetics ; immunology ; Neutralization Tests ; Phlebovirus ; genetics ; immunology ; Viral Proteins ; genetics ; immunology

Objective To evaluate the treatment of aneurysms rupture during endovascular embolization.Methods Nine aneurysms ruptured during the embolization and were treated with endovascular embolization.The reasons of aneurysms rupture during embolization,the prevention and the first aid after aneurysms rupture were analysed.Results Seven patients recovered and 2 died.Conclusions The optimal treatment of aneurysms rupture during endovascular embolization is effective,(J Intervent Radiol,2007,16: 132-134)

Objective To investigate the causes,consequences and management of injuries to the draining veins after embolization of brain arteriovenous malformations(BAVMs)with ?-n-butyl cyanoacrylate (NBCA).Methods The angiographic imaging data of 189 BAVMs patients who underwent NBCA embolization were studied retrospectively.The status of the draining veins before and after NBCA embolization was observed and compared.The intracerebral hemorrhage(ICH)complications and their relation to their angiographic features were analyzed.Results Twenty-three patients out of 189 patients showed injuries to the draining venous system,including 10 low-grade injury,6 moderate injury,and 7 high- grade injury.Six patients suffered from ICH after embolization,of whom 4 patients were due to injuries of the draining veins(2 moderate and 2 high-grade).In the 3 months follow-up evaluation of 4 patients with ICH, one died,one was in vegetative state,and the other two patients suffered from residual severe or minor (1 patient for each)permanent neurological deficits.Conclusion Our findings suggest that injury of the draining veins is the major cause of ICH and may lead to serious consequences after embolization of BAVMs with NBCA.

OBJECTIVETo study the effect of oleic acid (OA) on expression of aquaglyceroporin genes, AQP3 and AQP9, in hepatocyte steatosis and to investigate the underlying molecular mechanisms using an in vitro system.

METHODSHepG2 cells were treated with OA at different concentration to establish in vitro models of nonalcoholic hepatocyte steatosis. The corresponding extents of hepatic steatosis modeling were assessed by oil red O staining and optical density (OD) measurements of the intracellular fat content. The model lines were then treated with inhibitors of the PI3K/Akt and p38 MAPK signaling pathway factors and effects on AQP3/9 expression was measured by real time RT-PCR and western blotting.

RESULTSThe fat concentration, indicative of hepatic steatosis, increased in conjunction with increased concentrations of OA (0 less than 250 less than 500 mumol/L). OA exposure also down-regulated AQP3 mRNA and up-regulated AQP9 mRNA levels in a concentration-dependent manner. The most robust changes in expression occurred in response to the 500 mumol/L concentration of OA for both AQP3 (0.47+/-0.18; t = 4.5450, P less than 0.05) and AQP9 (1.57+/-0.21; t = 3.0306, P less than 0.05). Treatment with OA + PI3K pathway inhibitor (LY294004) significantly decreased AQP9 mRNA expression (4.55+/-0.62) as compared to the control group (1.00+/-0.10; t = 9.7909, P less than 0.01), that 500 mumol/L OA group (2.43+/-0.53; t = 4.5018, P less than 0.05), and the LY294002 group (1.90+/-0.16; t = 7.1683, P less than 0.01). Treatment with p38 MAPK pathway inhibitor (SB230580) significantly increased the OA-suppressed level of AQP3 mRNA to the level detected in the control group (1.27+/-0.11; t = 5.7455, P less than 0.01) and decreased the OA-stimulated AQP9 mRNA (0.38+/-0.09; t = 6.5727, P less than 0.01). No significant changes in mRNA expression of AQP3/9 were observed with inhibition of the ERK1/2 and JNK signal transduction pathways. The OA-induced changes in protein expression levels of AQR3 and AQP9 followed a similar trend of the genes. Finally, OA suppressed the level of phosphorylated Akt (from 0.21+/-0.02 to 0.13+/-0.03; t = 3.8431, P less than 0.05) but elevated the level of phosphorylated p38 (from 0.58+/-0.06 to 1.02+/-0.10; t = 12.5289, P less than 0.01). Again, OA treatment produced no significant affect on ERK1/2 and JNK phosphorylation.

CONCLUSIONOA down-regulates AQP3 expression by stimulating the p38 MAPK signaling pathway, and up-regulates the AQP9 by blocking the PI3K/Akt pathway and activating the p38 MAPK signaling pathway.

Aquaporin 3 ; metabolism ; Aquaporins ; metabolism ; Fatty Liver ; metabolism ; pathology ; Gene Expression Regulation ; drug effects ; Hep G2 Cells ; Humans ; Oleic Acid ; pharmacology ; Phosphatidylinositol 3-Kinases ; metabolism ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVE: To understand and analyze the morphological characteristics of hyoid bone and its gender difference in order to find out its forensic significance. METHODS: The hyoid bones of 68 adult corpses were dissected from redundant soft tissues after heating. The connection status of body of hyoid, greater cornu and lesser cornu, the morphological characteristics of hyoid bone and the degree of ossification were observed by visual inspection. The height of hyoid bone and the arched height of hyoid bone were measured and compared the differences between male and female in order to deduce the analytic equation for gender estimation by hyoid bone. RESULTS: Five connection conditions of hyoid bone were identified by the morphological characteristics, including complete ossification in both sides, no ossification in both sides, partial ossification in both sides, complete ossification in one side (partial ossification in the other side), and complete ossification in one side (no ossification in the other side). The values of the arched height of hyoid bone (x1) and the height of hyoid bone (x2) in male were both higher than that in female (P < 0.01). The analytic equation for gender estimation (y) was y = 0.438 x1 + 1.042x2-12.979. The discriminant value was -0.272 5 and the resolution was 88.2%. CONCLUSION According to the gender characteristics of hyoid bone, the data of hyoid bone can provide helps for forensic practices.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Aging/physiology* ; Body Size ; Cadaver ; Female ; Forensic Anthropology ; Humans ; Hyoid Bone/physiology* ; Male ; Middle Aged ; Osteogenesis ; Sex Characteristics ; Young Adult

OBJECTIVE: To find the mutation of disease-causing genes of sudden unexplained death syndrome (SUDS) in the young by whole exome sequencing in one case. METHODS: One SUDS case was found no obvious fatal pathological changes after conventional autopsy and pathological examination. The whole exome sequencing was performed with the Ion Torrent PGM™ System with hg19 as reference sequence for sequencing data. The functions of mutations were analyzed by PhyloP, PolyPhen2 and SIFT. A three-step bioinformatics filtering procedure was carried out to identify possible significative single nucleotide variation (SNV), which was missense mutation with allele frequency < 1% of myocardial cell. RESULTS: Four rare suspicious pathogenic SNV were identified. Combined with the analysis of conventional autopsy and pathological examination, the mutation MYOM2 (8_2054058_G/A) was assessed as high-risk deleterious mutation by PolyPhen2 and SIFT, respectively. CONCLUSION Based on the second generation sequencing technology, analysis of whole exome sequencing can be a new method for the death cause investigation of SUDS. The gene MYOM2 is a new candidate SUDS pathogenic gene for mechanism research.
Autopsy ; Brugada Syndrome/genetics* ; Cause of Death ; DNA Mutational Analysis/methods* ; Death, Sudden/etiology* ; Exome ; Gene Frequency ; Genetic Testing/methods* ; High-Throughput Nucleotide Sequencing/methods* ; Humans ; Molecular Biology ; Molecular Diagnostic Techniques/methods* ; Molecular Sequence Data ; Mutation

Objective To evaluate the effect ofendovascular embolization on intracranial aneurysms with detachable coil and detachable balloon and to emphasize the mainpoint of embolization technique.Methods 1328 patients underwent complete cerebral angiography using microcatheter under DSA imaging. 85 cases with 90 aneurysms were embolized by MDS, 825 cases with 847 aneurysms were embolized by GDC, 418 cases with 433 aneurysms were embolized by EDC. 37 of 67 giant aneurysms were embolized by detachable balloon, 18 aneurysms by GDC, 11 aneurysms by EDC, 1 by MDS. Results 1328 patients with 1370 aneurysms were cured successfully; 1 322 cases recovered clinically, 6 patients died. 1 281 aneurysms achieved 100 % occlusion, 65with 95 % occlusion, 20 with 90 % occlusion, and 4with 80 % embolization. 9 aneurysms reputured during the embolization, 5 patients had cerebral infarction, 1 patient died of intractable cerebral vasospasm for microcoil escape. 2 recurrent cases were cured by second GDC embolization. Conclusion The method of endovascular embolization to treat intracranial aneurysm is safe, reliable and effective. Those cases with giant aneurysms will have high recurrence. Patients who suffer from SAH repeatedly may have great possibility of aneurysm rupture during embolization.

Objective To study the short-term and long-term effectiveness of intracranial aneurysm occlusion with expandable hydrocoil. Methods Forty-one patients with intracranial aneurysms (n=45) were treated by endovascular occlusion with expandable hydrocoil. Follow-up interviews in the forms of DSA, CTA or MRA were conducted to the 41 patients within 6-24 months after the treatment to find out the tumor recurrence and complications. Results In the 41 patients, 1 died, 1 suffered from recurrence, 3 developed cerebral infarction, 1 got oculomotor paralysis, 2 got hydrocephalus. According to modified Rankin scale, grade 0 in 8 cases, grade 1 in 19, grade 2 in 7, grade 3 in 3, grade 4 in 2, grade 5 in 1 and grade 6 in 1. Conclusions Endovascular embolization with expandable hydrocoil is an effective treatment method for intracranial aneurysms, especially for parent artery occlusion, but it may be able to cause more complications in the treatment of small aneurysms (＜5mm), so the caution should be taken.

Objective To analyze the effect of NexusTM coils for endovascular occlusion of intracranial aneurysms. Methods In 41 patients with intracranial aneurysms, endovascular occlusion of 43 aneurysms was performed using NexusTM coils. The follow-up data of the patients for 6 to 12 months were reviewed, and the imaging data from digital subtraction angiography (DSA), CT angiography (CTA) or magnetic resonance arthrography (MRA) alter the treatment were analyzed. Results In the 41 patients, 1 died, 1 had aneurysm recurrence, 3 had cerebral infarction, 1 showed ocular paralysis, and 2 developed hydrocephalus after the surgery. Evaluation with modified Rankin Scale showed grade 0 in 8 cases, grade 1 in 19 cases, grade 2 in 7 cases, grade 3 in 3 cases, grade 4 in 1 case, grade 5 in 1 ease and grade 6 in 1 case. Conclusion Endovascular embolization with NexusTM coils is effective for treatment of intracranial aneurysms especially in cases of small aneurysms and parent artery occlusion. Caution should be taken with the coil for endovascular occlusion of the neck of anterior and middle cerebral artery aneurysms with thin parent arteries, as the fibers in the coil may cause thrombosis and potential cerebral infarction.

OBJECTIVETo establish a mouse model of humoral immune response by immunization with rabbit red blood cells (RRBCs).

METHODSThe mice were immunized with RRBCs and the serum hemolysin level was measured by micro-hemolysis spectrophotometry.

RESULTSThe peak time needed for hemolysin production against RRBCs was 6 days after the immunization, and 20% RRBCs in a total volume of 0.2 ml was optimal for intraperitoneal injection. Hydrocortisone (25 mg/kg) and cyclophosphamide (20 mg/kg) inhibited hemolysin production. Mannatide (4 mg/kg) produced no significant effect on serum hemolysin level in normal mice, but significantly potentiated hemolysin production in immunosuppressed mice induced by cyclophosphamide (20 mg/kg).

CONCLUSIONIntraperitoneal RRBC injection is feasible for establishing mouse models of humoral immune response.

Animals ; Erythrocytes ; immunology ; Female ; Guinea Pigs ; Hemolysin Proteins ; blood ; Immunity, Humoral ; Immunization ; Male ; Mice ; immunology ; Models, Animal ; Rabbits