Forty-two patients with type 2 diabetes mellitus (DM) were refered to 3 groups:type 2 DM without diabetic nephropathy (DN),type 2 DM with DN in the initial stage and type 2 DM with DN in the clinical stage.Ten healthy subjects were served as control group.Plasma adrenomedullin (ADM),urinary?_1- microglobulin (MG) and?_2-MG were detected.The results showed that the level of plasma ADM rose gradually with the development of DN and was positively correlated with markers of tubulointerstiial injury such as urinary?_1- MG and?_2-MG (both P

Humans ; Metals ; Microwaves ; adverse effects ; Occupational Exposure ; adverse effects ; analysis ; Workplace

Objective To investigate the efficacy and safety of the combined therapy of cyclophosphamide and thalidomide in the treatment of refractory Crohn's disease (CD).Methods This study was a prospective and open study.A total of 15 patients with refractory CD were enrolled.All patients received intravenous cyclophosphamide 200 mg every other day for two weeks,then followed by intravenous 400 mg once a week until the cumulative dose reached 6 to 8 g.when the cyclophosphamide treatment started,at the same time thalidomide was taken 25 to 75 mg every night according to the tolerance of patients.Before the treatment,two weeks' after the treatment and at the time when the cumulative dose of cyclophosphamide reached 6 to 8 g,Crohn's disease activity index (CDAI),hemoglobin (Hb),white blood cell (WBC) count,erythrocyte sedimentation rate (ESR) and high-sensitivity C-reactive protein (hs-CRP) were recorded.Endoscopy examination was conducted before the treatment and at the time when the cumulative dose of cyclophosphamide reached 6 to 8 g.The condition of mucosa healing was observed and scored by simple endoscopic score for crohn's disease (SES-CD).Adverse effects of all patients were monitored.Paired t test was performed for statistical analysis.Results Before the treatment,the CDAI of 15 patients with refractory CD was 235.87±59.87,two weeks after the treatment the CDAI declined to 135.33 ± 29.23,and the difference was statistically significant (t=7.50,P＜0.01).Before the treatment,ESR and hs-CRP was (42.13±22.80) mm/1 h and (13.73± 2.18) mg/L.Two weeks after treatment they declined to (23.80±16.63) mm/1 h and (5.77±4.77) mg/L,and the differences were statistically significant (t=2.43 and 6.17,both P＜0.05).After two-week treatment,10 patients achieved clinical remission.After the cumulative dose of cyclophosphamide reached 6 to 8 g combined therapy,CDAI of patients was 108.14 ± 47.10,which decreased significantly compared with that before treatment (t=6.30,P＜0.01).ESR,hs-CRP and WBC count was (19.35± 19.18) mm/1 h,(6.16± 5.02) mg/L and (6.28 ± 3.42) × 109/L,respectively,which decreased compared with those before treatment,and the differences were statistically significant (t=5.90,5.40 and 3.71,all P＜0.01).Twelve patients achieved clinical remission.And the lesions of 12 patients improved under endoscope,furthermore,the mucosa of four patients healed.Before the treatment,SES-CD was 9.14 ± 5.39,which declined to 5.07 ± 4.58 after the treatment,and the difference was statistically significant (t =3.14,P ＜ 0.01).During the treatment,five patients had adverse effects.Alanine aminotransferases (ALT) increased in three patients,WBC count decreased in one patient and one patient got a severe urinary infection.Conclusions Patients with refractory CD could achieve clinical remission,mucosa healing under endoscopy and better efficacy with the combined therapy of cyclophosphamide and thalidomide.However,adverse effects should be monitored during the treatment.

Artemisnin is a novel sesquiterpene lactone with an internal peroxide bridge structure, which is extracted from traditional Chinese herb Artemisia annua L. (Qinghao). Recommended by World Health Organization, artemisinin is the first-line drug in the treatment of encephalic and chloroquine-resistant malaria. In the present study, transgenic A. annua plants were developed by overexpressing the key enzymes involved in the biosynthetic pathway of artemisinin. Based on Agrobacterium-mediated transformation methods, transgenic plants of A. annua with overexpression of both HDR and ADS were obtained through hygromycin screening. The genomic PCR analysis confirmed six transgenic lines in which both HDR and ADS were integrated into genome. The gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had higher expression levels of HDR and ADS than the non-transgenic control (except ah3 in which the expression level of ADS showed no significant difference compared with control); and the HPLC analysis of artemisinin demonstrated that transgenic A. annua plants produced artemisinin at significantly higher level than non-transgenic plants. Especially, the highest content of artemisinin was found in transgenic line ah70, in which the artemisinin content was 3.48 times compared with that in non-transgenic lines. In summary, overexpression of HDR and ADS facilitated artemisinin biosynthesis and this method could be applied to develop transgenic plants of A. annua with higher yield of artemisinin.
Artemisia annua ; genetics ; metabolism ; Artemisinins ; metabolism ; Biosynthetic Pathways ; Drugs, Chinese Herbal ; Mixed Function Oxygenases ; genetics ; Oxidoreductases ; genetics ; Plant Proteins ; genetics ; Plants, Genetically Modified ; genetics ; metabolism ; Plants, Medicinal ; genetics ; metabolism

0.05).Conclusions SNP of 2350G→A in ACE gene is associated with MI,AA genotype is probably a genetic marker of MI in Han nationality.

Objectives To assess treatment outcomes and prognosis in infants with atrial flutter (AFL).Methods Thirty-four (34) cases of infants with AFL in Hunan Children's Hospital had been analyzed for clinical features, treatment outcomes and follow-up between March, 2009 and September, 2015. Based on ECG characteristics, the patients had been divided into simple and complex AFL groups. Based on age, they had been divided into neonates and non-neonates group. The aim of this study is to compare the clinical effects of drug treatment in different types of AFL.Resultsb With digitalis alone, the cardioversion rate was 37.5%,no signiflcant difference was observed between simple and complex AFL groups (45.8% vs 12.5%,P=0.206). Combining with other drugs, the cardioversion rate was 54.5%, which showed signiflcant difference between simple and complex AFL groups (76.9% vs 22.2%,P=0.036). The overall cardioversion rate was 70.6%, which showed signiflcant difference between simple and complex AFL groups(87.5% vs 30%,P=0.003). There was no signiflcant difference in pharmaceutical cardioversion rate between neonates and non-neonates group (85.7% vs 60.0%,P=0.216). Two cases with symptoms of heart failure used synchronized cardioversion. One patient restored to sinus rhythm, and another case was still recurrent of AFL after repeated electrical cardioversion, and eventually died of cardiogenic shock. After treatment, 9 patients were still with paroxysmal AFL and atrial tachycardia episodes, including 3 cases of simple type and 6 cases of complex type who were discharged with oral digoxin and propafenone treatment at home. 24 patients were followed up (3 months to 3 years and 4 months). 16 cases restored to sinus rhythm during hospitalization had no recurrence of AFL.Conclusions The overall treatment effects of AFL in infants were good. In simple type of AFL, most of patients did not need long-term antiarrhythmic drug therapy and the prognosis was good. The prognosis of treatment with conventional drug was poor in complex AFL group, with a higher rate of recurrence of AFL.

RhoA, a small GTPase, is involved in a wide array of cellular functions in the central nervous system, such as cell motility, cytoskeleton rearrangement, transcriptional regulation, phagocytosis and cell growth. It is not known how spinal cord injury (SCI) affects the expression of RhoA in different nerve cells. In the present study, we investigated the changes of RhoA expression in remote areas of the injury at the 3rd, 7th and 30th day after SCI, which was established by T10 contusion method. Moreover, we examine its expression profile in neurons, astrocytes and microglia. RhoA was found to be weakly expressed in these nerve cells in normal spinal cord. Western blotting showed that, after SCI, the total RhoA expression was up-regulated, and the RhoA expression was increased and peaked at the 7th day. Double immunostaining revealed specific and temporal expression patterns of RhoA in different nerve cells. The expression of RhoA in neurons started to increase at day 3, peaked at day 7 and then decreased slightly at day 30. Expression of RhoA in astrocytes increased moderately after SCI and peaked at day 7. There was no obvious change in RhoA expression in microglia after SCI in remote areas. This study demonstrated that, after SCI, RhoA expression exhibited different patterns with different nerve cells of spinal cord. RhoA expression patterns also changed with time after SCI, and among different nerve cells in the injured spinal cord. These findings can help us better understand the roles of RhoA in SCI.

Peptide cyclization, a pivotal approach to modifying linear precursors of proteins and pepticles, has been used to enhance their biological activities and serum stabilities. Recently, sortase A (SrtA) from Staphyloccus aureus becomes a promising new technology for efficiently incorporating site specific modifications into proteins, conjugating the cell surface and cyclizing the linear peptides. In this study, we constructed two recombinant expression systems, one with chitin binding domain and the other with six-histidine tag and chitin binding domain on the N-terminal of SrtA, separately. The results of enzymatic kinetics indicate that the two recombinant tags do not impair the transpeptidase activity of SrtA compared with the standard reaction reported under the same reaction condition. The two synthesized peptides with N-ternimal three glycines and C-terminal penta-amino acid motif, LPETG, were cyclized using immobilized and recycled SrtA. The SrtA-based cyclization promises to represent a simple method for easy and efficient enzymatic synthesis of large cyclic peptides.
Aminoacyltransferases ; metabolism ; Bacterial Proteins ; metabolism ; Cyclization ; Cysteine Endopeptidases ; metabolism ; Enzymes, Immobilized ; metabolism ; Kinetics ; Peptides ; metabolism ; Peptides, Cyclic ; biosynthesis ; Staphylococcus aureus ; enzymology

OBJECTIVETo construct a eukaryotic expression plasmid pcDNA3.1(-)-Humanin.

METHODSThe recombinant plasmid pGEMEX-1-Humanin was digested with restriction endonucleases BamH I and Hind III and the Humanin gene fragments, about 100 bp length, were obtained. Then the Humanin gene fragments were inserted into eukaryotic expression vector pcDNA3.1(-) and the recombinant plasmids pcDNA3.1(-)-Humanin were identified by sequencing.

RESULTSRecombinant plasmid DNA successfully produced a band which had the same size as that of the Humanin positive control. The sequence of recombinant plasmids accorded with the Humnain gene sequence.

CONCLUSIONSA eukaryotic expression plasmid of Humanin was successfully constructed.

Base Sequence ; Cloning, Molecular ; methods ; Escherichia coli ; genetics ; metabolism ; Humans ; Intracellular Signaling Peptides and Proteins ; Molecular Sequence Data ; Molecular Weight ; Plasmids ; genetics ; Protein Engineering ; methods ; Proteins ; chemistry ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; chemistry

Artemisnin is a novel sesquiterpene lactone with an internal peroxide bridge structure, which is extracted from traditional Chinese herb Artemisia annua L. (Qinghao). Recommended by World Health Organization, artemisinin is the first-line drug in the treatment of encephalic and chloroquine-resistant malaria. In the present study, transgenic A. annua plants were developed by overexpressing the key enzymes involved in the biosynthetic pathway of artemisinin. Based on Agrobacterium-mediated transformation methods, transgenic plants of A. annua with overexpression of both HDR and ADS were obtained through hygromycin screening. The genomic PCR analysis confirmed six transgenic lines in which both HDR and ADS were integrated into genome. The gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had higher expression levels of HDR and ADS than the non-transgenic control (except ah3 in which the expression level of ADS showed no significant difference compared with control); and the HPLC analysis of artemisinin demonstrated that transgenic A. annua plants produced artemisinin at significantly higher level than non-transgenic plants. Especially, the highest content of artemisinin was found in transgenic line ah70, in which the artemisinin content was 3.48 times compared with that in non-transgenic lines. In summary, overexpression of HDR and ADS facilitated artemisinin biosynthesis and this method could be applied to develop transgenic plants of A. annua with higher yield of artemisinin.