The Proteomic Research of the Cure of Experimental Diabetes Deafness by Granules of Eliminating Phlegm and Removing Blood Stasis

Objective: Observing the expression changes of serum proteome in model rats after intervention of the Granules of Eliminating Phlegm and Removing Blood Stasis (豁痰祛瘀颗粒 also known as GEPRB), screening out and identifying the differentially expressed proteins by mass spectrometry and bioinformatics analysis, discussing the molecular mechanism of control the Diabetes deafness by GEPRB. Methods: By use of proteomics technology, the serum protein serum proteome of the control group, model control group, Duxil and each observation group were observed for 2-DE gel pattern matching, and the difference in the relative content of 2 times was chosen for the differentially expressed proteins. Identification of differentially expressed proteins by MALDI-TOF MS/MS, the authors further analysis the phosphorylation, subcellular localization, interaction, direct regulation, and transmembrane of the differences proteins by the way of bioinformatics analysis. Sixty SPF level SD rats elected in diabetic rats model group (abbreviated as DM group) were be randomly divided into 5 groups based on random number sheet, namely model control group, positive drug control group (Du-ke-xi group) and Mai-tong-fang high, medium and low dose group respectively. In addition, set of normal control group. 10 rats in each group. Results: By Coomassie brilliant blue staining, identified 51 differential protein spots dug from 2-D gel by mass spectrometry, successfully identified 13 non-redundant proteins. Most of the identified proteins were secreted protein and belong to different protein families. There were about 12 proteins have the transmembrane region from the authors’ result, ten of them were plasma membrane proteins. Conclusion: It’s suggesting that 13 differential proteins is most likely the protein response to GEPRB in vivo, these proteins may play key role for the treatment of GEPRB to Diabetes deafness. The two highly differentially expressed proteins Apolipoprotein E (apoE) and C3 may be a potential drug target of GEPRB.

Heteroplasmy Level of the Mitochondrial tRNA^(Leu(UUR)) A3243G Mutation in a Chinese Family Is Positively Associated with Earlier Age-of-onset and Increasing Severity of Diabetes

Objective To investigate the mutations of mitochondrial genome in a pedigree with suspected maternally inherited diabetes and deafness and to explore the correlations between the mutations and clinical features. Methods Genomic DNA was isolated from blood leucocytes of each member of the pedigree. The mitochondrial genome was amplified with 24-pair primers that could cover the entire mitochondrial DNA. Direct sequencing of PCR products was used to identify any mitochondrial DNA mutations. Results Family members on the maternal side all harbored the tRNA^Lcu（UUR） A3243G mutation. The paternal side family members did not have the mutation. The age-of-onset of diabetes of the 4 maternal side family members was 15, 41, 44, and 65 years old, and their corresponding heteroplasmy level of the mutation was 34.5%, 14.9%, 14.6%, and 5.9%, respectively. The age-of-onset of diabetes and heteroplasmy level of A3243G mutation were negatively correlated with a correlation coefficient of -0.980（P=0.02）. Meanwhile, patient with high heteroplasmy level of A3243G mutation had relatively low severity of disease. Moreover, 6 reported polymorphisms and 2 new variants were found. Conclusions The main cause of diabetes in this pedigree is the tRNA^Lcu（UUR） A3243G mutation. However, other gene variants may contribute to its pathogenicity. The heteroplasmy level of the tRNA^Lcu（UUR） A3243G mutation is positively associated with earlier age-of-onset and increasing severity of diabetes.

Effect of hypoglycemic anti-deafness capsules in diabetic patients with deafness and toxicological assessment in rats

OBJECTIVE:Through experiment on animals and clinical trials to explore the safety and efficacy of hypoglycemic anti-deafness capsules on diabetic patients with deafness.METHODS:Total 296 patients with non-insulin dependent diabetes mellitus(NIDDM)were randomly divided into two groups.A treatment group of 164patients(208 ears)was treated with hypoglycemic anti-deafness capsules based onTCM syndrome differentiation.A control group of 132 patients(184ears)was treated with glibenclamide and conventional drug treatment for deafness.The following were observed:hearing,fasting plasma glucose(FPG),2 h postprandial plasma glucose(2hPG),24 h urine glucose(24hUG),improvement of main symptoms,platelet function,and changes in superoxidedismutase(SOD)and lipid peroxide(LPO)levels.In animal studies,Kunming mice,weighing 18-22 g were used.Half of the mice were males and half were females.Wistar rats,weighing 80-120 g were used.Half of the rats were males and half were females.Male Wistar rats,weighing 200-220 g,were also used.Their acute and chronic toxicity was studied.RESULTS:The hearing improvement was 56.7%in the treatment group and 26.6%in the control group.FPG,2hPG,and 24hUG were improved significantly(P<0.05,P<0.01,P<0.01,respectively)in the treatment group and 2hPG and 24hUG improved significantly in the control group(P<0.05,P<0.05).The improvement in 2hPG and 24hUG in the treatment group was significantly greater than that in the control group P<0.01).There was no significant difference in FPG between the two groups(P<0.05).Main symptoms in the treatment group were significantly more improved than those in the control group(P<0.05,P<0.01).In the treatment group,platelet adhesion and aggregation,SOD,and LPO were all significantly improved from before treatment(P<0.05,P<0.01).However,in the control group,except LOP(P<0.05),there were no significant differences from before treatment to after(P<0.05).In animal studies,no obvious acute or long-term toxicity was observed from capsule administration.CONCLUSION:Through experiment on animals and clinical trials,we can found that hypoglycemic anti-deafness capsules could decrease blood glucose and serum triglycerides of alloxan-induced diabetic rats.This herbal capsule is effective for safely treating diabetic patients with deafness.