ObjectiveTo explore the efficacy and safety of percutaneous antegrade stenting in the treatment of ureteral obstruction after renal transplantation.MethodsWe retrospectively reviewed 11patients with renal graft ureteral obstruction (2 cases of acute obstruction and 9 cases of chronic obstruction) from March 2009 to March 2011.The etiology of the obstruction was renal graft-ureter-bladder anastomotic stricture in 5 cases,stone obstruction in 2 cases,and undetermined in 4 cases.Renal graft and collecting system were examined by ultrasonography preoperatively to select suitable puncture position,and then ureteropyelography was performed under X-ray guidance.When the obstruction location was clear,the urology guidewire was implanted to the bladder by needle,and then guidewire was released by cystoscopy.Ureteral stent was implanted along the guidewire,and upper ureteral stents was observed under X-ray. After removal of guidewire,the stent location was confirmed once again.The renal pelvis fistula drainage lasted for 1-2 weeks,and ureteral stent to 6 months to one year.Ultrasound and renal function were tested after 1week,1month,3 months and 6 months,and then every six months.ResultsOperation was done successfully in 10 patients,and failed in one case due to a long segment of ureteral stenosis.The operating time of ureteral stent implantation was 54±27 min.Serum creatinine of patients was reduced from preoperative 326±147 to postoperative 89±49 μmol/L.During a follow-up period of 6 to 27 months,no complications occurred.ConclusionPercutaneous antegrade stenting in the treatment of ureteral obstruction after renal transplantation is safe and effective.

Objective To investigate the method to isolate and culture hepatic stellate cells ( HSCs) for studying the cellular mechanisms of hepatic frbrosis.Methods HSCs were isolated by nycodenz density gradient centrifugation after the hepatocytes obtained from adult male gebils were digested with pronase, collagenase and DNase, infused via portal vein.The cell viability was determined by trypan blue exclusion test.The purity of HSCs was identified by detectingα-SMA, desmin immunohistochemical staining.Results The yield rate of HSCs was 0.5~1 ×107 per gerbil liver, and the cell viability was more than 90%.The percentage ofα-SMA-positive cells was more than 75%after 3 days primary culture and almost 100% cells were α-SMA and desmin positive in passage culture.Conclusion The successful protocol of primary culture of Mongolian gerbil HSC provide a technical support for research of relevant liver diseases and drug development in the future.

Objective To study the relationship between rs290487,rs7903146 of transcription factor 7-Like 2 (TCF7L2) gene and post-transplantation diabetes mellitus in Han Zhejiang population.Method We genotyped two single nucleotide polymorphisms (SNPs) across the TCF7L2 gene in 90 unrelated post-transplantation type 2 diabetes mellitus (PTDM) patients,112 unrelated non-PTDM patients,and a set of post-transplantation diabetes mellitus patients (n =68).Genotyping was performed using direct sequencing SNP Genotyping Assays.The association of SNPs with post-transplantation diabetes mellitus was analyzed.Result In this study,there was statistically significant difference in the T-allele of TCF7L2.rs7903146 between PTDM group (5.1％) and non-PTDM group (1.3％) (P＜0.05).For rs7903146,the frequencies of genotype C/C,C/T (70.0％) and T/T (35.8％) was statistically significant in PTDM group (P＜0.05).For rs290487,the frequencies of genotype C/C,C/T and T/T was 14.7％,38.2％ and 47.1％ respectively in PTDM group,P＞ 0.05.The incidence of PTDM was significantly higher in patients with the CT genotype (odds ratio 18.54 [95％ CI 1.21-282.26],P =0.03).Conclusion With the current sample size,we found that the CT genotype of rs7903146 was significantly associated with post-transplantation diabetes mellitus.

Objective To explore the correlation between post-transplant glomerular filtration rate (GFR) in 1 year and long-term graft survival in renal transplant patients.Methods The clinical data of 334 patients who received their cadaveric kidney transplantations between November 1994 and October 2004 were analyzed retrospectively.According to the GFR at one year after transplant operation, normal GFR group was defined as GFR more than or equal to 1.083 ml/s, while patients whose GFR less than 1.083 ml/s were fallen into abnormal GFR group.Cockeroft-Gault (C-G) formula was used to compare the difference in the renal function between the two groups.Kaplan-Meier assay was used to compare the difference in the allograft survival between the two groups in the functional renal allograft or the non-functional renal allograft.The correlativity of GFR level at the first year and the GFR level at the 5th year was analyzed.Results The GFR level at the first year after transplantation was proportional to the graft survival time of the kidney.Five and ten years after transplantation, the renal transplantation long-term survival rate in the normal renal function groups was significantly higher than in the abnormal renal function groups (P＜0.05).As compared with the GFR level at the first year after transplantation, the changes in amplitude of GFR level at the 5th year after transplantation was (0.080 ±0.248) ml/s, and the descent had a positive correlation with GFR level at the 5th year after transplantatioa Conclusion GFR level at the first year after transplantation predicts long-term renal functioa The higher of GFR level at the first year, the higher of GFR level at the 5th year.

Sepsis is an organ dysfunction that endangers a patient's life caused by an imbalanced infection response, and is a clinically critical illness. Despite a deep understanding of the pathogenesis of sepsis, there has been no significant improvement in sepsis mortality during clinical treatment at home and abroad. In recent years, the role of autophagy in the pathogenesis of sepsis has become a new research point in the field of medical research. Autophagy may protect the body by removing pathogenic microorganisms, neutralizing microbial toxins, and regulating cytokine release in sepsis. Studies have shown that autophagy plays a role in heart and lung organ dysfunction and inflammatory immune response in sepsis. Studies have also shown that hydrogen sulphide (H 2S) can activate autophagy through multiple signaling pathways, such as adenylate-activated protein kinase/mammalian target of rapamycin (AMPK/mTOR), phosphoinositide 3 kinase/Akt/mTOR (PI3K/Akt/mTOR), liver kinase B1/STE20 related adapter protein/mouse protein 25 (LKB1/STRAD/MO25) and microRNA-30c (miR-30c), etc. signaling pathways. This article reviewed the effects of H 2S on autophagy-related genes Beclin-1 and microtubule-associated protein light 3 chain (LC3) on intestinal function of sepsis in order to explore the H 2S-mediated autophagy gene expression in pus. The protective role of autophagy gene for intestinal dysfunction provides a new strategy for the treatment of sepsis in the future.

Objective To analyze the clinical features of nonimmunocompromised patients with allergic bronchopulmonary aspergillosis (ABPA).Methods The clinical data of 11 nonimmunocompromised patients diagnosed as ABPA from June 2010 to December 2015 in Zhejiang Jinhua People's Hospital were retrospectively analyzed.SPSS 18.0 was used for analysis.Results Among 11 patients with ABPA, Five were males and 6 were females, with an average age of (49.3 ±11.0) years.All patients had cough, expectoration and wheezing;cough and tan sputum in 4 cases, bloody sputum in 3 cases, fever in 2 cases and chest pain in 2 cases.In auscultation dry rales were heard in all patients , and limited wet rales were heard in 3 cases.The peripheral blood leukocyte counts were elevated in 5 patients [11.7(10.3-13.5) × 109/L)] and the eosinophils counts were increased in 9 patients [1.79(0.09-7.63) ×109/L].The total IgE was elevated to 3640(1329-9430) IU/mL.Skin prick test was positive ( grade 3 to 5) in 10 cases, Aspergillus fumigatus specific IgE increased to 23.6(1.75-67.30) kU/L in 6 cases, Aspergillus fumigatus specific IgG raised to 83.3(51-126) mg/L in 5 cases.Chest CT showed patchy, punctate exudation in 8 cases, central bronchiectasis in 9 cases, bronchial mucosal plug formation in 4 cases, and atelectasis in 1 case.Mediastinal lymph nodes were found in 2 cases.All 11 patients were treated with glucocorticoid hormone, and 8 patients were also received itraconazole oral solution for treatment.After treatment, the clinical symptoms were improved rapidly.Conclusion Nonimmunocompromised patients with ABPA have no specific clinical manifestations , and often are misdiagnosed as asthma , which is worth the attention of clinicians.

Objective Cysteinyl leukotrienes are potent inflammatory mediators. Their actions are mediated by specific receptors,the CysLT receptors(CysLT1R and CysLT2R),which have been cloned and characterized. In this stud-y,we investigated the protective effects of the CysLTR antagonist Pranlukast and HAMI 3379 on global cerebral ischemia/reperfusion(CI/R)injury in gerbils and its underlying mechanisms. Methods The gerbil model of CI/R was established by bilateral common carotid artery occlusion for 10 min followed by 24 h reperfusion. Then the animals were equally ran-domized into four groups: sham, model, Pranlukast(0.1 mg/kg)and HAMI 3379(0.1 mg/kg)groups. The later two groups were treated with intraperitoneal injection of Pranlukast and HAMI 3379,respectively,once daily for 4 days before carotid artery occlusion,while the former two groups with saline only,all at 10 mL/kg. After 24 h reperfusion,neurologi-cal deficit scores were observed and the behavioral dysfunction was assessed. The neuron morphology of cerebral cortex and CA1 subregion of hippocampus were observed in brain sections stained with cresyl violet. The expression of autophagy-relat-ed proteins beclin-1 and LC3 in the homogenate of cerebral cortex and hippocampus were determined using western blotting analysis. The ultrastructure of autophagosomes in the CA1 subregion of hippocampus was observed by electron microscopy. Results Compared with the model group, Pranlukast and HAMI 3379 attenuated neurological deficits, improved the be-havioral dysfunction,inhibited the neuron injury and loss, decreased the expression of autophagy-related protein beclin-1 and LC3 and the number of autophagosomes. Conclusions cysteinyl Leukotriene receptor antagonist Pranlukast and HAMI 3379 can alleviate global cerebral ischemia/reperfusion injury in gerbils. The protective effects of Pranlukast and HAMI 3379 appear to be associated with the inhibition of autophagy.

Gram-negative Enterobacteriaceae with resistance to carbapenem conferred by New Delhi metallo-β-lactamase 1 (NDM-1) are a type of newly discovered antibioticresistant bacteria. The rapid pandemic spread of NDM-1 bacteria worldwide (spreading to India, Pakistan, Europe, America, and Chinese Taiwan) in less than 2 months characterizes these microbes as a potentially major global health problem. The drug resistance of NDM-1 bacteria is largely due to plasmids containing the blaNDM-1 gene shuttling through bacterial populations. The NDM-1 enzyme encoded by the blaNDM-1 gene hydrolyzes β-lactam antibiotics, allowing the bacteria to escape the action of antibiotics. Although the biological functions and structural features of NDM-1 have been proposed according to results from functional and structural investigation of its homologues, the precise molecular characteristics and mechanism of action of NDM-1 have not been clarified. Here, we report the three-dimensional structure of NDM-1 with two catalytic zinc ions in its active site. Biological and mass spectroscopy results revealed that D-captopril can effectively inhibit the enzymatic activity of NDM-1 by binding to its active site with high binding affinity. The unique features concerning the primary sequence and structural conformation of the active site distinguish NDM-1 from other reported metallo-β-lactamases (MBLs) and implicate its role in wide spectrum drug resistance. We also discuss the molecular mechanism of NDM-1 action and its essential role in the pandemic of drug-resistant NDM-1 bacteria. Our results will provide helpful information for future drug discovery targeting drug resistance caused by NDM-1 and related metallo-β-lactamases.
Amino Acid Sequence ; Anti-Bacterial Agents ; metabolism ; Binding Sites ; Captopril ; chemistry ; pharmacology ; Catalytic Domain ; Crystallography, X-Ray ; Drug Resistance, Bacterial ; Enterobacteriaceae ; enzymology ; Molecular Sequence Data ; Sequence Alignment ; Sequence Homology, Amino Acid ; beta-Lactamases ; chemistry ; metabolism

Arabidopsis BOTRYTIS-INDUCED KINASE1 (BIK1) is a receptor-like cytoplasmic kinase acting early in multiple signaling pathways important for plant growth and innate immunity. It is known to form a signaling complex with a cell-surface receptor FLS2 and a co-receptor kinase BAK1 to transduce signals upon perception of pathogen-associated molecular patterns (PAMPs). Although site-specific phosphorylation is speculated to mediate the activation and function of BIK1, few studies have been devoted to complete profiling of BIK1 phosphorylation residues. Here, we identified nineteen in vitro autophosphorylation sites of BIK1 including three phosphotyrosine sites, thereby proving BIK1 is a dual-specificity kinase for the first time. The kinase activity of BIK1 substitution mutants were explicitly assessed using quantitative mass spectrometry (MS). Thr-237, Thr-242 and Tyr-250 were found to most significantly affect BIK1 activity in autophosphorylation and phosphorylation of BAK1 in vitro. A structural model of BIK1 was built to further illustrate the molecular functions of specific phosphorylation residues. We also mapped new sites of FLS2 phosphorylation by BIK1, which are different from those by BAK1. These in vitro results could provide new hypotheses for more in-depth in vivo studies leading to deeper understanding of how phosphorylation contributes to BIK1 activation and mediates downstream signaling specificity.
Amino Acids ; chemistry ; Arabidopsis ; enzymology ; Arabidopsis Proteins ; chemistry ; genetics ; isolation & purification ; Gene Expression Regulation, Plant ; Immunity, Innate ; Mutation ; Phosphorylation ; Protein-Serine-Threonine Kinases ; chemistry ; genetics ; isolation & purification ; Signal Transduction ; Threonine ; genetics

Severe fever with thrombocytopenia syndrome virus (SFTSV), a member of the Phlebovirus genus from the Bunyaviridae family endemic to China, is the causative agent of life-threatening severe fever with thrombocytopenia syndrome (SFTS), which features high fever and hemorrhage. Similar to other negative-sense RNA viruses, SFTSV encodes a nucleocapsid protein (NP) that is essential for viral replication. NP facilitates viral RNA encapsidation and is responsible for the formation of ribonucleoprotein complex. However, recent studies have indicated that NP from Phlebovirus members behaves in inhomogeneous oligomerization states. In the present study, we report the crystal structure of SFTSV NP at 2.8 Å resolution and demonstrate the mechanism by which it processes a ringshaped hexameric form to accomplish RNA encapsidation. Key residues essential for oligomerization are identified through mutational analysis and identified to have a significant impact on RNA binding, which suggests that correct formation of highly ordered oligomers is a critical step in RNA encapsidation. The findings of this work provide new insights into the discovery of new antiviral reagents for Phlebovirus infection.
Binding Sites ; Crystallography, X-Ray ; Mutation ; Nucleocapsid Proteins ; chemistry ; genetics ; metabolism ; Phlebovirus ; metabolism ; Protein Binding ; Protein Multimerization ; Protein Structure, Quaternary ; RNA, Viral ; metabolism ; Recombinant Proteins ; biosynthesis ; chemistry ; genetics