ObjectiveTo generate the prokaryotic expression systems of vwA1 and vwA2 genes of Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai,and to determine the correlation among the target recombinant expressed products rVwA1 and rVwA2 and platelet-associated hemorrhage.MethodsThe vwA1 and vwA2 genes of L.interrogans strain Lai were amplified using high fertility PCRs and then the amplification products were sequenced.The prokaryotic expression systems of vwA1 and vwA2 genes were generated by using routine genetic techniques.The expression and dissolubility of rVwA1 and rVwA2 proteins were examined by SDS-PAGE plus Bio-Rad Gel Image Analyzer.Ni-NTA affinity chromatography was used to extract the expressed rVwA1 and rVwA2.Real-time fluorescence quantitative RT-PCRs were performed to determine the changes of vwA1-mRNA and vwA2-mRNA levels in the leptospires of L.interrogans strain Lai before and after infection of human umbilical vein endothelial cell line (HUVEC).The ability of leptospiral rVwA1 binding to human platelets was detected by flow cytometry.A hydrolytic test plus SDS-PAGE were applied to examine the cleavage of leptospiral rVwA2 by a human von Willebrand factor lysase (ADAMTS13).ResultsThe nucleotide and putative amino acid sequences of the cloned leptospiral vwA1 and vwA2 genes were 100％ identities compared to the reported corresponding genes.The generated prokaryotic expression systems of vwA1 and vwA2 genes could express soluble rVwA1 and rVwA2,respectively.When L.interrogans strain Lai infected HUVEC for 8 h,both the vwA1-mRNA and vwA2-mRNA levels were significantly up-regulated (P＜0.05).The result of flow cytometry showed that the leptospiral rVwA1 was able to combine with human platelets with a 60.8％ binding rate. Human recombinant vWF-A2 ( rhvWF - A 2 ),but not the leptospiral rVwA 2,could be hydrolyzed by human ADAMTS 13.Conclusion The vwA1 and vwA2 genes may play roles during infection of L.interrogans species,and the function of vwA1 gene product is referring to the hemorrhage in leptospirosis.

Child, Preschool ; Female ; Humans ; Infant ; Male ; Rotavirus Infections ; diagnosis ; therapy ; Vomiting ; etiology

Objective To explore the efficacy and safety of bedside continuous blood purification (CBP) in the treatment of neonatal multiple organ failure (MOF).Methods Totally 6 newborn infants of MOF were hospitalized in department of neonatology in our hospital from June 2011 to June 2013.These 6 cases of clinical data were retrospectively analyzed,6 neonates were treated with CBP combined with conventional treatment.The model for CBP was continuous veno-venous hemodialysis filtration (CVVHDF),blood flow velocity was 3 to 5 ml/(kg· min),replacement fluid dose was 20 to 30 ml/(kg· h),dialysis fluid dose was 15 to 25 rnl/(min· m2).The clinical outcome measures included,blood pressure,blood pH,K+,Na+,blood urea nitrogen,creatinine,urine volume,PaO2/FiO2 and epinephrine intravenous dose,respectively before CBP treatment,6 h,12 h,24 h,48 h after CBP treatment and the end of CBP treatment.The efficacy of CBP treatment was evaluated in neonatal MOF.Results Gestational age of 6 neonates with MOF was 33 to 41 weeks,2 to 19 days old,2.25 to 3.36 kg birth weight.Primary disease was 4 cases of neonatal septicemia(1 case with congenital hereditary metabolic disease),2 cases of severe neonatal asphyxia.All 6 cases of venous catheter were smoothly done.CBP treatment persisted for 49 to 106 hours.Compared with before CVVHDF treatment,blood K+,blood urea nitrogen,creatinine significantly decreased at 12 h after CVVHDF treatment [(5.32 ± 1.84) mmol/L vs.(9.81 ±3.61) mmol/L,(9.0 ±3.4) mmol/L vs.(12.8 ±6.1) mmol/L,(99 ± 16) μmol/L vs.(176 ±25) μmol/L,P ＜0.05],and reached the normal range at 24 h after treatment,urine volume significantly increased at 24 h after treatment (P ＜ 0.05).PaO2/FiO2 reached 200 mmHg (1 mmHg =0.133 kPa) at 6 h after treatment and more than 300 mmHg at 24 h after treatment(P ＜0.05).Fifty percent of epinephrine intravenous dose were down-regulation at 12 h after treatment and stopped using epinephrine at 48 h after treatment.CBP treatment of 6 cases showed effective.Conclusion Application of bedside CBP treatment in neonatal MOF is safe,can effectively help neonates with MOF to skip over renal failure stage.

Mesenchymal chondrosarcoma originated in the primitive mesenchymal tissue .It usually devel-ops in the short bones such as hand ,foot and body bone ,while extremeIy rare in the orbit .We report a case of mesenchymal chondrosarcoma of the orbit which is confirmed by pathology .

Background:MicroRNA?506 (miR?506) has been reported to function in several tumors as a tumor suppressor gene or oncogene. However, the expression and role of miR?506 in pancreatic ductal adenocarcinoma (PDAC) remains unclear. In this study, we aimed to evaluate the phenotype of miR?506 in PDAC. Methods:Using miRNA insitu hybridization, we examined the expression of miR?506 in 113 PDACs and 87 paired normal pancreatic tissues. We evaluated miR?506 expression in PDAC cells, normal pancreatic ducts, and acinus/islands, and we analyzed the associations between miR?506 expression and the clinicopathologic characteristics of PDAC patients. Results:miR?506 expression was higher in PDAC than in matched normal pancreatic ductal cells (P<0.001). On the other hand, the combined group of well and moderately differentiated PDACs showed higher levels of miR?506 than the poorly differentiated ones (P=0.023). Moreover, miR?506 expression was negatively associated with pathologic T category (P=0.004) and lymph node metastasis (P=0.033), suggesting that miR?506 might inhibit the progression of PDAC. Conclusions:Our results suggest that miR?506 either plays a role as an oncogene in the tumorigenesis and a tumor suppressor in the progression or serves as a house?keeping, tumor?suppressing miRNA, whose expression can be activated by oncogenic signals in early development to hinder the progression of PDAC.

AIM To investigate the protection of plicamycin on apoptosis in cerebellar granule neurons (CGN) of rat. METHODS TUNEL, Hoechst 33258 staining, agarose gel electrophoresis and fluorescein diacetate staining were used to detect morphological and biochemical characteristics of apoptosis in primary rat CGN. RESULTS Being pre-incubated with plicamycin for 1 h and lasting for 24 h, rat CGN apoptosis induced by low potassium basal modified Eagle′s medium for 24 h was inhibited in a plicamycin concentration-dependent manner. This effective concentrations of plicamycin were from 50 to 200 nmol·L-1, and the maximum inhibitory rate of plicamycin on CGN apoptosis was near 80% at 200 nmol·L-1. CONCLUSIONPlicamycin inhibits rat CGN apoptosis induced by low potassium.

Adolescent ; Adult ; Aged ; Bronchi ; Child ; Female ; Foreign Bodies ; surgery ; Humans ; Male ; Middle Aged ; Surgical Mesh ; Trachea ; Young Adult

OBJECTIVETo investigate the relationship between chronic pyelonephritis (CPN) and immune function, and the therapeutic mechanism of Yishenkang granule (YSKG).

METHODSOne hundred and twenty patients of CPN were divided into 3 groups randomly, the YSKG group (treated with YSKG), the control A group (treated with Sanjin tablet) and the control B group (treated with western medicine). Serum levels of immunoglobulin (Ig), complement 3 (C3), interleukin-2 (IL-2), peripheral T-lymphocyte subsets, and urinary secretory immunoglobulin A (sIgA) were determined before and after treatment with monoclonal antibody assay, agar diffusion method, radioimmunoassay (RIA), and radioimmuno-equilibrium method, and compared with normal control.

RESULTSThere were disorders of T-lymphocyte subsets in CPN, lowering of serum Ig, C3 and urinary sIgA, and increase of blood IL-2 (P < 0.05 or P < 0.01). These abnormalities could be normalized after YSKG treatment.

CONCLUSIONFunctional disorders of cellular and humoral immunity exist in CPN patients of chronic stage, YSKG could correct the immune functional disorder, control the recurrence of CPN effectively and alleviate the immunopathological damage.

Adult ; Chronic Disease ; Complement C3 ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Immunoglobulin G ; blood ; Interleukin-2 ; blood ; Male ; Middle Aged ; Pyelonephritis ; drug therapy ; immunology ; prevention & control ; T-Lymphocyte Subsets ; drug effects

This study is to investigate the protective effect of rosiglitazone (RSG) against learning and memory impairment of APP/PS1/tau transgenic mice. AD mice model was replicated by using 6-month APP/PS1/tau transgenic mice. The learning and memory ability of mice was evaluated by Morris water maze and Western blotting assays was applied to measure the phosphorylation and O-glycosylation of Tau and neurofilaments (NFs) protein. The results demonstrated that RSG could reverse the learning and memory deficits of 3 x Tg mice significantly. It was also found that RSG could suppress the hyperphosphorylation of Tau and NFs protein levels and increase the glycosylation expression of Tau and NFs proteins in 3 x Tg mice brain. Together, RSG ameliorates cognitive impairments of 3 x Tg mice via the alleviation of the hyperphosphorylated Tau and NFs proteins burden in the brain.
Alzheimer Disease ; Amyloid beta-Peptides ; Animals ; Brain ; drug effects ; Disease Models, Animal ; Glycosylation ; Learning ; drug effects ; Memory ; drug effects ; Memory Disorders ; drug therapy ; Mice ; Mice, Transgenic ; Neurofilament Proteins ; metabolism ; Phosphorylation ; Thiazolidinediones ; pharmacology ; tau Proteins ; metabolism

【Objective】To study the effect of the specific p38 mitogen-activated protein kinase(p38 MAPK) inhibitor SB203580 on apoptosis of cerebellar granule neurons induced by low potassium.【Methods】Apoptosis was induced by switching the cultured cerebellar granule neurons from a culture medium containing K+ 25 mmol*L-1 to a medium containing K+ 5 mmol*L-1 (cLK).Fragmentation of DNA was analyzed using agarose gel eletrophoresis.SAPK/JNK activity was measured by SAPK/JNK assay kit.【Results】Low potassium resulted in apoptosis as characterized by morphological and biochemical features,but the specific p38 MAPK inhibitor SB203580 improved the survival of cerebellar granule neurons cultured in cLK medium by blocking apoptosis in a concentration-dependent manner.The expression and phosphorylation of c-Jun increased and the activity of c-Jun N-terminal protein kinase (JNK) elevated when cerebellar granule neurons were cultured in cLK medium.But when the cerebellar granule neurons cultured in cLK medium were exposed to 25 μmol*L-1 SB203580,the expression and phosphorylation of c-Jun and the activity of JNK were both decreased evidently.【Conclusions】These results indicate that SB203580 inhibits the activation of JNK and phosphorylation of c-Jun,and therefore protects granule neurons from apoptosis induced by low potassium.