2.Global adiponectin suppress the high expression of monocyte chemotactic protein-1 induced by high glucose in NRK52E cells
Tao YAO ; Xiaoyan WU ; Wenli YU ; Ping GAO ; Yizhe WU
Chinese Journal of Nephrology 2015;31(2):145-149
Objective To investigate the effect of globular adiponectin on the high expression of monocyte chemotactic protein-1 (MCP-1) induced by high glucose in rat renal tubular epithelial cells (NRK52E),and its relationship with adiponectin receptors and p38MAPK.Methods NRK52E cells were cultured in vitro and divided into six groups:normal glucose group (NG,5.6 mmol/L glucose),high glucose group(HG,25 mmol/L glucose),gAd groupl (HG+gAd 2 mg/L),gAd group2 (HG+gAd 5 mg/L),gAd group3 (HG+gAd 10 mg/L),p38MAPK antagonist group:(SB,HG+SB203580 10 μmol/L).The protein expression of phosphorylated p38MAPK (p-p38MAPK),total p38MAPK (t-p38MAPK),MCP-1 and AdipoR1/AdipoR2 were examined by western blotting.The mRNA expression of MCP-1 and AdipoR1/AdipoR2 were detected by RT-PCR and real-time PCR respectively.Results Compared with NG group,the mRNA and protein expression of MCP-1 increased significantly in HG group (all P< 0.05).The phosphorylation of p38MAPK increased (P< 0.05) with no change in t-p38MAPK protein.The addition of gAd or SB203580 inhibited the unregulation of MCP-1 and p-p38MAPK induced by HG.Two kinds of adipoR,adipoR1 and adipoR2,were all detectable in NG group,and mRNA and protein expression of adipoR1 was higher than that of adipoR2 (P< 0.01).Compared with NG group,the expression of adipoR decreased in HG group,but the difference had no statistical significance(P > 0.05).Compared to HG group,the mRNA and protein expression of adipoR1 increased in gAd groups (all P < 0.01).Conclusion The gAd can dose-dependently attenuate the overexpression of MCP-1 induced by high glucose,and this protective effect may be mediated by adipoR1 and p38MAPK.
3.Diagnostic and monitoring values of peripheral blood cardiac troponin Ⅰmessager RNA for myocardial damnification
Jianhua ZHU ; Dengfu YAO ; Wei WU ; Zengdong GAO ; Gongsheng SHI
Chinese Journal of Tissue Engineering Research 2005;9(39):158-161
BACKGROUND: Cardiospecific proteins of the troponin-tropomyosin complex in the contractile system of the cardiomyocytes have challenged creatine kinase isoenzyme MB (CK-MB) as the "gold standard" for the early biochemical detection of acute myocardial injury.OBJECTIVE: To investigate cardiac troponin Ⅰ messager RNA (cTnI-mRNA) in peripheral blood and its clinical values in diagnosis of patients with myocardial injury.DESIGN: A basic and observational study for set up a method to analyze cTnI-mRNA.SETTING: Department of Cardiology, Affiliated Hospital, Nantong University.MATERIALS: The project was accomplished from May 2003 to May 2005 in Research Center of Clinical Molecular Biology, and Department of Pathology, Affiliated Hospital, Nantong University. The cTnI-mRNA was detected from blood by a nested PCR assay, and its clinical values as a sensitive myocardial diagnostic marker were confirmed in patients with myocardial injury.METHODS: Pathologic features and microstructure of cardiac myocytes were examined by H&E staining or electron microscopy. The cTnI-mRNA was extracted from blood and synthesized to cDNA through random primers and reverse transcriptase, and amplified by a nested PCR assay, and its clinical values as a myocardial diagnostic marker were investigated in patients with myocardial injury.MAIN OUTCOME MEASURES: Microstructure of cardiomyocytes, sensitivity of analysis method and diagnostic values.RESULTS: Microstructure of cardiomyocytes with mitochondria swell,rupture, vacancy-like denaturation, nucleus abnormality, and chromatin condensed were observed by electron microscopy. The cTnI-mRNA fragments from heart and blood were successfully amplified and the sensitivity was 2 pg/μL. The product sequences from tissues or blood were confirmed by sequencing. The cTnI-mRNA from cardiac myocytes was found that it present in blood plasma and not in circulating nucleus cell. The incidence of blood cTnI-mRNA of chronic cardiomyopathy was significantly higher (P < 0.05) than that of serum enzymatic patterns or cTnI quality,respectively.CONCLUSION: The analysis of blood cTnI-mRNA is a sensitive marker for diagnosis and monitoring of myocardial injury.
4.Protein levels and its clinical significance of septin-9 and clusterin in peripheral blood of epithelial ovarian cancer patients
Nenan LYU ; Hongwen YAO ; Ting XIAO ; Yanning GAO ; Lingying WU
Chinese Journal of Obstetrics and Gynecology 2015;(9):679-684
Objective To evaluate septin-9 and clusterin protein levels in the peripheral blood samples from epithelial ovarian cancer patients, and explore its clinical significance. Methods Clinical data of 200 patients in Cancer Hospital,Chinese Academy of Medical Sciences from Jan. 29, 2008 to Feb. 1,2010 were collected. The peripheral blood samples were obtained from 137 epithelial ovarian cancer patients, 12 borderline ovarian tumor patients, 10 benign ovarian tumor patients, 41 benign pelvic lesion patients and 58 healthy women. The septin-9 and clusterin protein levels in the plasma were measured by double antibody sandwich ELISA or ELISA. The clinical significance of clusterin and septin-9 in plasma was analyzed. The diagnostic efficacy of septin-9 and clusterin protein in the detection of ovarian cancer was evaluated by the area under the curve (AUC) of the receiver operating characteristic (ROC) curve. Results Double antibody sandwich ELISA showed: the mean levels of plasma septin-9 in epithelial ovarian cancer patients or benign pelvic lesion patients were significantly higher than that in healthy women detedted by double antibody sandwich ELISA (P<0.01). The mean levels of plasma septin-9 in epithelial ovarian carcinoma patients with tumor family history or distance metastasis were significantly higher than those patients without (P<0.05). While the expression level of septin-9 protein in peripheral blood of ovarian cancer patients was not related to the patient age, pathologic stage, pathologic differentiation, smoking history, treatment history (including surgery, radiotherapy and chemotherapy) and lymph node metastasis (all P>0.05). ELISA showed: the mean level of plasma clusterin in epithelial ovarian cancer patients was significantly higher than that in healthy women deteded by ELISA (P=0.021). The expression level of clusterin protein in peripheral blood of ovarian cancer patients was not related to the above clinical pathological parameters (all P>0.05). To distinguish between ovarian cancer patients and healthy women by septin-9 protein expression level in plasma, when AUC was 0.712 and cut off was 0.28, the sensitivity of detection ovarian cancer by septin-9 protein expression was 82.5%, and the specificity was 50.0%. To distinguish between ovarian cancer patients and healthy women by clusterin protein expression level in plasma, when AUC was 0.636 and cut off was 87.96 pg/L, the sensitivity of detection ovarian cancer by clusterin protein expression was 71.5%, and the specificity was 41.4%. Conclusions The expression of septin-9 and clusterin protein in peripheral blood of ovarian cancer patients is increased, especially the expression level of septin-9 protein with related to the distant metastasis. The study results shown that the detection of septin-9 and clusterin in plasma has a certain diagnosis value in ovarian cancer, which may be a potential markers for ovarian cancer.
5.Diagnostic value of array-based single nucleotide polymorphisms comparative genomic hybridization in An-gelman syndrome
Jing GAO ; Xiyu HE ; Yao YANG ; Honglin WU
Chinese Journal of Applied Clinical Pediatrics 2015;(18):1401-1404
Objective To analyze the genotype-phenotype correlations of Angelman syndrome ( AS ) , and to discuss the advantage of applying array-based single nucleotide polymorphisms comparative genomic hybridization ( SNP aCGH) in diagnosis of AS. Methods Examination of electroencephalogram( EEG) and intelligence quotient( IQ) evaluation were done for 11 cases diagnosed as AS clinically. Gesell scares were chosen as the evaluation criterion of IQ. The screening techniques was methylation polymerase chain reaction( MS-PCR) ,then SNP aCGH was used to make genetic diagnosis. Results (1)Eleven cases of AS were confirmed:1 case had UPD(uniparental disomy),10 cases were type of deletion, from which 6 cases were deletion (Ⅱ) , 4 cases were deletion (Ⅰ) . ( 2 ) The copy number variations were detected in the region of 15q11-q13,which contained genes like MKRN3,MAGEL2,NDN,SNRPN, SNURF,GABRB3,GABRA5,GABRG3,UBE3A,OCA2,ATP10A. To search online Mendelian inheritance in man,genes above were correlated with AS manifestation. (3)All cases of deletion were 3-5 standard deviation(SD) in weight and height to normal children at the same age and with the same sex,while UPD was below 1. 5 SD. Gesell scares showed that the deletion(Ⅰ) was the most serious in mental retardation,deletion(Ⅱ) was moderate,and the UPD was mild. Eight cases were hypopigmentation,and one was the UPD. EEG revealed that 1 case of deletion(Ⅰ) and the UPD were spike occasionally,another one deletion(Ⅰ) was limit EEG. The rest cases displayed slow and spike waves paroxysmal-ly,with amplitude of medium or high,2. 5-3. 0 Hz. Conclusions Not only can SNP aCGH make a diagnosis of AS but discriminate the types of genetic pathology. Since different type contributes to a diverse of clinical features and the rate of recurrence is also different,it is significant for family genetic consultation. Moreover,the technology is advantageous for the study on the pathogenesis and gene function.
6.Study of copy number variations in children with unexplained mental retardation/brain development delay
Jing GAO ; Yao YANG ; Honglin WU ; Xiyu HE
Chinese Journal of Applied Clinical Pediatrics 2016;31(20):1550-1555
Objective By using array-based single nucleotide polymorphisms comparative genomic hybridization (SNP-aCGH) to detect and fine mapping copy number variations (CNVs) in children with unexplained mental retardation/brain development delay(MR/BD),then the CNVs were analyzed to determine diagnosis and offer genetic counseling,finally to discuss the application of SNP-aCGH in genetic diagnosis of MR/BD with unknown causes.Methods Ninety-two children with unexplained MR/BD were recruited.SNP-aCGH was used to get CNVs from the whole genome-wide,and the correlation of CNVs and phenotype was analyzed to definit disease genes or pathogenic fragment.Statistics was performed to analyze the common phenotype between positive cases (case with CNVs) and negative cases.Results (1) The CNVs were detected in 10 cases with a detection rate of 10.86%,from which 8 cases showed subtelomeric aberration,5 cases without subtelomeric aberration,and the rate was 8.70%,5.40%,respectively.The CNVs related to MR/BD involved 10 different subtelomeric regions (9p,21q,3p,2p,15q,4p,12p,22q,16p,17p),and 7 different regions without subtelomeric (1p,4q,2p,14q,15q,12q,22q).The deletions involved 11 zones (size:1.05-8.80 Mb),and duplications referred to 8 zones (size:1.33-31.25 Mb).(2) One case was diagnosed as 9p duplication syndrome,for candidate genes:DOCK8,VLDLR.A case was detected with a gene fracture (CRBN).One case was diagnosed as Coffin-Sirrs syndrome combined with a deletion of 15q26.3-qter,for candidate genes:SOX11 and LINS1,respectively.One case referred to 12p13.3 deletion syndrome,for candidate genes:ELKS,ERC1.One case referred to 22q13.2-qter deletion,for candidate genes:SHANK3.Two cases were diagnosed as ATR-16 syndrome with 17p13.3 deletion syndrome,their candidate genes:HBA1,HBA2,SOX8 for the former,YWHAE,LIS1 for the latter.(3)There were statistically significant differences in comparison of positive cases to the negative ones for growth delay,internal organs deformity,low birth weight infant(LBW) and premature infant (all P <0.05).Conclusions (1) Besides MR/BD in different degrees in all the positive cases,they also showed growth delay,a portion of them with internal organs deformity,low birth weight infant and premature infant.(2) Subtelomeric aberrations are related to MR/BD,while the submicroscopic rearrangement in regions without subtelomeric is suspiciously pathogenic,and need to be further studied.(3) SNP-aCGH can fine mapping the region of CNVs by high resolution from the whole genome-wide,which does contribute to limit the zones for finding pathogenic region and candidate genes,as well as to offer a technology platform for investigating about the correlation of phenotype and genes or CNVs.
7.Infection after cardiac-lung transplantation in ICU
Ping GAO ; Zhengxian CHEN ; Hua YAO ; Ruobin WU ; Keli HUANG
Chinese Journal of Practical Internal Medicine 2000;0(12):-
Objective To explore the characteristics of infection after cardiac-lung transplantation.Methods March 23th 2006,one patient received orthotopic heart-lung transplantation,the clinical data were observed and analyzed.Results Depend on etiology,antibioti and antifungul were selected,the patient recoved.Conclusion It is very important to early etiology diagnosis and reasonable selection of antibiotic.
8.The analysis of blood transfusion and hospitalization duration between the occlusion of ventricular septal defect through transesophageal echocardiography and the repair of ventricular septal defect
Yao YAO ; Li XIE ; Xin WANG ; Qin WU ; Lei GAO ; Yifeng YANG
Journal of Chinese Physician 2013;(2):186-189
Objective To compare the rate of blood transfusion and hospitalization duration between the occlusion of ventricular septal defect through transesophageal echocardiography and the repair of ventricular septal defect.Methods A total of 131 consecutive patients were included in the study from July of 2011 to December of 2011,75 patients of them received the occlusion of ventricular septal defect through transesophageal echocardiography,and 56 patients of them received the repair of ventricular septal defect.Results All of 131 patients had discharged without death.The group of receiving the repair of ventricular septal defect presented the higher rate of blood transfusion(x2 =81.83,P < 0.01) and longer hospitalization duration than the other(t =46.57,P < 0.01).Conclusions The group of receiving the occlusion of ventricular septal defect through transesophageal echocardiography had the lower rate of blood transfusion and shorter hospitalization duration.
9.Determination of Emodin and Danshensu in Pinggan Oral Liquid by HPLC
Yang WU ; Xu SUN ; Yuan GAO ; Xingdong LI ; Xiaoyu MA ; Huifeng YAO ; Yao CHEN
China Pharmacist 2016;19(4):758-760,761
Objective:To establish the method for the determination of emodin and danshensu Pinggan oral liquid. Methods:The contents were determined by HPLC. The chromatographic conditions for emodin were as follows: an Agilent Eclipse XDB-C18 column (150 mm × 4. 6 mm, 5μm) was used and the mobile phase was methanol-water (80 ∶20). The detection wavelength was 287 nm and the flow rate was 1. 0 ml·min-1 . The column temperature was 25℃ and the injection volume was 20 μl. The chromatographic condi-tions for Danshensu were as follows:an Agilent Eclipse SB-C18 column (150 mm × 4. 6 mm, 5 μm) was used and the mobile phase was methanol-1% acetic acid solution (2 ∶98). The detection wavelength was 280 nm and the flow was 1. 0 ml·min-1. The column temperature was 30℃ and the injection volume was 20 μl. Results:Emodin had a good linear relationship within the range of 26. 75-428. 00 μg·ml-1(r=0. 999 9). The average recovery was 102. 7% and RSD was 0. 76% (n=6). Sodium Danshensu had a good linear relationship within the range of 10.17-162.77 μg·ml-1(r =0.999 9). The average recovery was 100.5% and RSD was 1. 47% (n=6). Conclusion:The method is simple, accurate and reproducible, which can be used for the quality control of the prep-aration.
10.Efficacy and safety of harmonic scalpel in neck dissection: a Meta-analysis.
Yao YAO ; Yehai LIU ; Kaile WU ; Chaobing GAO ; Yi ZHAO ; Jing WU ; Yifan LI ; Yang WANG ; Tao WANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(10):915-920
OBJECTIVE:
To systematically evaluate the efficacy and safety of harmonic scalpel in neck dissection.
METHOD:
Available literatures of PubMed, EMBASE, Cochrane Library, Google Scholar, CBM, CNKI, WangFang and VIP published before June 2014 were searched. Inclusion criteria and quality assessment were performed. All data were analyzed by using RevMan 5.2 software.
RESULT:
Fourteen studies including 632 cases were enrolled. Among them, 319 cases were in harmonic scalpel group and 313 cases in conventional resection group. Compared with conventional resection group, the harmonic scalpel group showed shorter surgery time(weighted mean difference [95% confidence intetval]: -28.01 [-36.83, -19.19], Z = 6.22, P < 0.01)and less intra-operative blood loss (weighted mean difference [95% confidence intetval]: -46.68 [-57.25, -36.12], Z = 8.66, P < 0.01). The number of cervical lymph nodes dissected and the incidence of postoperative chylous leakage were similar in both groups.
CONCLUSION
Using the harmonic scalpel in neck dissection was as efficient and safe as that of the conventional technique with the advantage of shorter time of surgery and less intraoperative blood loss.
Blood Loss, Surgical
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Humans
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Lymph Nodes
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Neck
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surgery
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Neck Dissection
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instrumentation
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Postoperative Period
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Surgical Instruments