BACKGROUND: Vasorelaxation plays an important role In the occurrence of cyclosporine A (CsA)-induced nephrotoxlcity.OBJECTIVE: To observe the alleviative effects of green tea polyphenols (GTP) on CsA-induced inhibition of vasorelaxation and the underlying mechanisms.METHODS: Sprague-Dawley rats were randomly and evenly divided into four groups: CsA, control, CsA + GTP, and GTP. After 5 weeks of drug treatment, blood urea nitrogen (BUN) and creatinine (Cre) levels were determined. Then the thoracic aorta rings were mounted on a bath system, and acetylcholine was used to induce vasorelaxation. The effects of L-NAME and indomethacin and the denuded vasorelaxation were evaluated.RESULTS AND CONCLUSION: The BUN and Cre levels in the CsA group were significantly higher than those in the control group (P < 0.05). The maximal response (Emax%) for acetylcholine-induced vasorelaxation in the CsA group was significantly lower than that in the control and GTP groups. After pretreatment with L-NAME, vasorelaxation was significantly lower in the CsA,CsA+GTP and GTP groups than in the control group. After pretreatment with indomethacin, vasorelaxation was significantly higher in the control, CsA +GTP, and GTP groups than in the CsA group. The level of nitric oxide metabolites in the vascular tissue in the CsA group was significantly lower compared with other groups. The results demonstrated that CsA can decrease nitric oxide levels in vascular tissues and induce abnormal endothelium-dependent vasorelaxation, which is mediated by nitric oxide pathway.

Objective To study the optimal dose and reaction time of human leukocyte antigen B27(HLA‐B27)antibody in flow cytometry .Methods Take 52 cases of whole blood in patients with ankyl‐osing spondylitis(AS) .According to HLA‐B27 antibody doses ,samples were divided into two groups:5 μL group and 10 μL group .HLA‐B27‐positive rate were tested after 5 ,10 ,15 min , respectively .Results The HLA‐B27 positive rate of 5 μL group at different reaction time were (84 .16 ± 1 .21)% ,(94 .81 ± 1 .33)% ,(94 .10 ± 1 .26)% ;the positive rate of 10 μL group at different reaction time were (85 .40 ± 1 .27)% ,(96 .76 ± 1 .31)% , (95 .36 ± 1 .45)% .The positive rate of HLA‐B27 in 10 μL group was higher than 10 μL group(F=90 .08 ,P<0 .05) .The positive rate of HLA‐B27 after reacting for 10 and 15 min were higher than that after reacting for 5 min(F=60 .25 ,P<0 .05) .There was not statistically significantly different between the reaction time of 10 min and 15 min(F=1 .08 ,P>0 .05) .Conclusion The opti‐mal dose and reaction time of HLA‐B27 antibody in flow cytometry are 10μL and 10 min;There is not any interaction between anti‐body dose and the reaction time of HLA‐B27 antibody .

Objective To evaluate the effects of fructose-1,6-diphosphate (FDP) on the preservation and high-energy phosphate metabolism of hypothermic SD rat hearts. Methods Sixteen SD rats were randomly divided into 2 groups. Their hearts were harvested and preserved in UW solution at 4℃ for 12 hours. In the study group (n=8), FDP (5mmol/L) was added to the UW solution, whereas in the control group, FDP was not added. Langendorff perfusion was established with the isolated hearts, and cardiac functions were examined, as well as the activities of lactate dehydrogenase (LDH) and creatine kinase (CK) in the perfusion fluid collected from coronary. Then the high-energy phosphate concentration, ATP, ADP, and AMP contents in the myocardium were determined with HPLC. Results The high-energy phosphate concentration was remarkably higher in the study group than that in the control group (P

Objective To investigate the expressions of stern cell markers CD133,nestin and CD44 in malignant melanoma and their significance.Methods Tissue samples were obtained from 30 patients with malignant melanoma and 30 patients with intradermal nevus.The expressions of three markers were immunohistochemically detected in the samples.Results In malignant melanoma specimens,the expression rate of CD133,nestin and CD44 was 53.33%(16/30),80.00%(24/30)and 20.00%(6/30),respectively,significantly difierent from that in intradermal nevus specimens [23.33%(7/30),53.33%(1 6/30)and 0,respectively,all P＜0.05].The percentage of cells positive for CD133,nestin and CD44 was 2.98%±5.62%,34.92%±34.89%and 1.28%±3.26%,respectively,in malignant melanoma specimens.0.10%±0.21%,7.26%±13.13%and 0,respectively,in intradermal nevus specimens;there was a significant difierence between the two groups of specimens(all P＜0.05).In malignant melanoma and intradermal nevus,the expression intensity of CD133.nestin and CD44 showed no significant correlation with patients'sex.age or disease course(all P＞0.05).ConclusionsCD133,nestin and CD44 are highly expressed in malignant melanoma,but weakly expressed or absent in intradermal nevus,suggesting that tumor stem cells might exist in malignant melanoma tissue.

Objective To evaluate the effect of conversion from mycophenolat mofetil (MMF) to enteric-coated mycophenolate sodium (EC-MPS) on gastrointestine-related quality of life,as well as the safety and efficacy.Method A total of 41 renal transplant (RT) recipients were converted from MMF to EC-MPS (46.3 ± 17.1) months after the operation due to the gastrointestinal side effects of MMF,with a mean time of 46.3 months.Before the conversion and 12 weeks later,the patients were evaluated with Gastrointestinal Quality of Life Index (GIQLI) questionnaire,and the safety and efficacy were assessed.Result The average dose of MMF was 846.7 ± 291.3 mg/day before the conversion,with a mean dose of 639.5 ± 186.4 mg/day for EC-MPS.The total score of GIQLI was 103.6 ± 10.7 before the conversion,and 12 weeks after conversion 118.3 ± 15.1,with a statistical significance (P＜0.05).The safety of EC-MPS was excellent without infection,acute rejection episode,loss of allograft or death.The serum creatine was 136.9 ± 35.7 mol/L before conversion and 128.4± 40.8 mol/L after conversion (P ＞ 0.05).Conclusion For the RT patients with gastrointestinal side effects of MMF,conversion to EC-MPS could significantly alleviate gastrointestinal illness,and improve quality of life,with excellent safety and efficacy.

Objective? To?investigate?the?antimicrobial?resistance?and?beta-lactamase?production?profile?in?the?gram-negative?bacilli?isolated from urinary tract infections in the Second People's Hospital of Gansu Province during the period from 2014 to 2015. The results will provided to clinicians for better antimicrobial treatment. Methods? The?bacterial?isolates?were?identified?via?conventional?laboratory?tests?or?automatic?identification?systems?and?subjected?to?antimicrobial?susceptibility?testing?by?using?Kirby-Bauer?method.?Three-dimensional test was used to detect the enzymes conferring antimicrobial resistance. The susceptibility testing results were interpreted according to the CLSI breakpoints issued in 2012. Results A total of 987 gram-negative strains were isolated from urinary tract infections, including E. coli (51.6%, 509/987), Enterobacter (11.0%, 109/987), P. aeruginosa (10.5%, 104/987), K. pneumoniae (9.9%, 98/987), P. mirabilis (9.3%, 92/987), C. freundii (4.7%, 46/987), and other gram-negative bacilli (2.9%, 29/987). ESBLs were produced?in?494?(50.1?%)?of?the?987?strains?of?gram?negative?bacilli.?Preliminary?screening?test?identified?243?AmpC?beta-lactamases?producing?strains,?and?135?(13.7?%)?strains?were?confirmed?by?three-dimensional?test.?Both?ESBLs?and?Amps?beta-lactamases?were?produced in 16 (1.6%) strains. The prevalence of ESBLs-producing strains was 79.6% in E. coli, 34.6% in P. aeruginosa and 37.8% in K. pneumoniae?isolates.?Metallo-β-lactamase?or?KPC?beta-lactamase?was?not?identified.?The?antimicrobial?resistance?was?serious?in?gram-negative bacilli, which showed relatively low resistance rate to imipenem (0.02%), amikacin (10.6%), cefoperazone-sulbactam (23.8 %), and nitrofurantoin (25.2 %). E. coli prevalence of levofloxacin,?ciprofloxacin?resistance?respectively?were?89.8%,?91.8%. The gram-negative bacilli from urinary tract also showed various levels of resistance to cephalosporins. Conclusions The gram-negative bacilli isolated from urinary tract infections are mainly E. coli. The gram-negative isolates show high level antimicrobial resistance and high prevalence of beta-lactamases. Imipenem, amikacin, cefoperazone-sulbactam, and nitrofurantoin still have very high antibacterial activity against these isolates in vitro.

AIM: To obtain GST fusion protein of hSP17 gene and construct the recombinant plasmid for expression in E. coli. METHODS: Total fragment of hSP17 cDNA gene were amplified by RT-PCR, then subcoloned into pGEX-3b to generate recombinant hSP17/pGEX. Right orientation of insert are identified by restricted enzyme digestion. Transform the correct recombinant plasmid into the E. coli DH5a. The expression of fusion proteins hSP17-GST were induced by adding isopropylthiogalactoside (IPTG). RESULTS and CONCLUSION: The recombinant plasmid hSP17/pGEX-3b could express effectively in E.coli and a high level of fusion protein hsp17-GST with the predicted molecular weight was detected.

Animals ; Bronchi ; Child ; Child, Preschool ; Foreign Bodies ; Humans ; Leeches ; Male

Here we discuss whether N terminal sequencing is appropriate as one of the conventional control methods for monoclonal antibody products. We determined the N terminal sequences of two monoclonal antibody products targeting two antigens separately with both Edman degradation and mass peptide spectrometry. We also identified the characteristic peptide fragments with mass spectrometry. Furthermore, we analyzed their heterogeneity with ion exchange chromatography, capillary zone electrophoresis and Imaged Capillary Isoelectric Focusing. Edman degradation method showed that the N terminal 15 amino acids of heavy and light chains of the two monoclonal antibodies were identical. Peptide mass spectrometry demonstrated that T1 peptide fragments of heavy and light chains of the two antibodies were also the same. But in contrast, peptide mapping and the three analytical methods for heterogeneity analysis could effectively identify and differentiate the two antibodies. The N terminal sequences of two monoclonal antibodies are identical because the number of framework sequences of humanized or human monoclonal antibodies is relatively limited, so whether N terminal sequencing analysis could be regulated as one of the practical control methods should be carefully discussed. Our work also proves that the above analytical methods could combinatorially applied to the identification of monoclonal antibody products, and are more objective compared to N terminal sequencing.
Amino Acid Sequence ; Antibodies, Monoclonal ; isolation & purification ; Chromatography, Ion Exchange ; Humans ; Isoelectric Focusing ; Mass Spectrometry ; Peptide Mapping ; Peptides ; Sequence Analysis, Protein ; methods

Objective To investigate the effect of using pterional approach combined with partial orbital roof and zygomatic process resection (modified orbitopterional approach)for the management of ruptured anterior communicating artery aneurysms. Methods From October 2013 to October 2016,36 consecutive patients with ruptured anterior communicating artery aneurysm admitted to the Department of Neurosurgery,Binzhou Medical University Hospital were enrolled retrospectively. They were all confirmed by DSA or CT angiography. The orbitopterional approach was used,only part of the orbital roof was removed and the structures of lateral orbital wall and the wings of sphenoid bone were not removed. The clinical manifestations,imaging data,surgical methods,and surgical results of ruptured anterior communicating artery aneurysms were summarized. Results All 36 patients with anterior communicating artery aneurysm in this group were treated with the modified orbitopterional approach. There was no obvious brain retraction injury on CT scan after procedure. At the time of discharge,the Glasgow outcome scale score was 5 in 25 cases, 4 in 8 cases,and 3 in 3 cases. No patients died. The patients were followed up for 3 to 24 months;no rebleeding and recurrence were observed. No complications occurred,such as enophthalmos,damage to the eyeballs, and cranial nerve injury. Conclusion The modified orbitopterional approach increased the operation space,avoided the distraction of brain tissue,significantly shortened the operation distance,and increased the deep observation angle through the removal of part of the orbital roof and the zygomatic process. It is suitable for the treatment of anterior communicating artery aneurysms,especially the rear direction,upper direction and high positioned aneurysms. There was no bone loss in the procedure,no need for orbital roof reconstruction. The gyri rectus was retained anatomically,and it may be helpful to protect the postoperative cognitive function of the patients.