Objective To construct the pGEX-4T-2-CblN/Grb2, express and purify the corresponding chimeric protein; To investigate whether the chimeric CblN/Grb2 possesses ubiquitin ligase activity. Methods Total RNA of SK-BR-3 cells were isolated and reversely transcribed into cDNA, which were used as templates to amplify Grb2 SH2 by PCR. The gene fragment encoding the N-terminal part of Cbl protein (named as CblN) was amplified by PCR using pEFHACbl plasmid encoding human Cbl as templates. BamH I and EcoR V restriction enzyme digestion sites were introduced into both flanks of SH2 by overlapping extension PCR and the modified gene were cloned into pcDNA3.1(+). The pcDNA3.1(+)-CblN/Grb2 was obtained by replacing SH2 of CblN with Grb2SH2 and then used as templates to amplify CblN/Grb2 by PCR. The pGEX-4T-2-CblN/Grb2 was constructed by subcloning CblN/Grb2 into the prokaryotic expressing vector pGEX-4T-2. The GST-CblN/Grb2 fusion protein was expressed in E. coli of DH5? under IPTG induction and further purified with Glutathione Sepharose 4B. In vitro ubiquitination assay was performed to investigate whether the GST-CblN/Grb2 fusion protein is able to mediate auto-ubiquitinating reaction, namely whether it possesses ubiquitin ligase activity. Results The fusion expressing vector of pGEX-4T-2-CblN/Grb2 was successfully constructed; The GST-CblN/Grb2 fusion protein was correctly expressed and purified; In vitro ubiquitination assay indicated that GST-CblN/Grb2 fusion protein is able to mediate auto-ubiquitinating reaction and therefore possesses ubiquitin ligase activity. Conclusion Expression, purification of GST-CblN/Grb2 and identification of its′ activity have laid the foundation for further study of chimeric ubiquitin ligase′ effects on the growth of HER2 positive tumor cells.

Objective To explore the clinical significance of urine transferrin testing in the diagnosis of early pregnancy-induced hypertension syndrome (PIH).Methods Sixty-eight PIH patients (PIH group) who received treatment from January 2012 to October 2013 were enrolled in this study.Mild PIH was 21 patients,midrange PIH was 24 patients and severe PIH was 23 patients.Meanwhile,68 cases of healthy person were selected as control group.The level of urine transferrin,urea and creatinine were compared between two groups.Results The level of urine transferrin,urea and creatinine in PIH group were significantly higher than those in control group[(3.01 ± 1.82) mg/L vs.(1.29 ± 0.88) mg/L,(13.12 ± 5.51) mmol/L vs.(6.23 ± 3.68) mmol/L,(135.5 ± 62.3) μ mol/L vs.(85.1 ± 38.2) μ mol/L] (P ＜ 0.05).The sensitivity of urine transferrin,urea and creatinine in mild PIH was 76.19％,61.90％,57.14％.In midrange PIH was 75.00％,79.17％,75.00％.In severe PIH was 78.26％,86.96％,73.91％.The specificity of urine transferrin,urea and creatinine was 80.65％,56.45％,58.06％.The sensitivity of different degree of PIH was exceeded 75.00％.The sensitivity of urine transferrin in mild PIH was significantly higher than urea and creatinine (P ＜ 0.05).But the sensitivity of urine transferrin in midrange PIH and severe PIH had no significant difference (P ＞ 0.05).The specificity of urine transferrin was significantly higher than urea and creatinine (P ＜ 0.05).Conclusions Urine transferrin is a good index in diagnosis of early PIH,which can reflect the early injury degree of the renal tubular and glomerular.It has active and important significance for the clinical diagnosis and treatment of PIH.

Administration, Oral ; Anti-Inflammatory Agents, Non-Steroidal ; adverse effects ; therapeutic use ; Aspirin ; adverse effects ; therapeutic use ; Brain Edema ; chemically induced ; Child ; Coma ; chemically induced ; Female ; Humans ; Liver ; drug effects ; pathology ; physiopathology ; Reye Syndrome ; chemically induced ; Treatment Outcome

Zoonoses are a class of infectious diseases that are transmitted from animals to humans. More than 200 known types of zoonoses have been reported across the world until now. Among 1 400 pathogens of human infectious diseases, approximately 61% are zoonotic origin, and 75% human emerging infectious diseases are zoonoses. These zoonoses pose a great threat to human and animal health and decrease livestock production. To effectively tackle the persistent challenges resulting from zoonoses, WHO collaborates with member governments, academia, non-governmental and charitable organizations, and regional and international partners to prevent and manage zoonotic threats and their public health, social and economic impacts. Although great success has been achieved in the management of zoonoses, there are still multiple challenges for zoonoses control in China due to environmental, climate, socioeconomic factors and antimicrobial resistance. Based on the One Health concept, the integration of modern biological, information, artificial intelligent and big data tools through multidisciplinary and multi-sectorial collaborations may facilitate the containment and elimination of zoonoses.

Objective To explore the relationship of the ADAM33 gene Q-1,T2 single nucleotide polymorphism (SNP) and suffering from chronic obstructive pulmonary disease (COPD) in Xinjiang Kazakh and Han population. Methods Peripheral blood samples to extract DNA, and the single nucleotide polymorphisms of Q-1 and T2 in ADAM33 gene were detected by SNaPshot SNP genotyping. Results Case group compared with the control group, frequencies of Q-1 locus genotypes and alleles were significant differences in Kazak (P<0.05). In patient group, there were significant differences in ADAM33 gene Q-1 locus genotypes FEV1% predicted, FEV1/FVC of clinical indicators lung function in Kazak, Han(P < 0.05). Kazak Q-1 locus AA genotype,Han GA genotype than GG genotype were significant difference.Compare Kazak AA genotype,Han GA genotype with GG genotype is more likely to cause COPD in Q-1 locus(P<0.05). In the comparison of the case and control group the two ethnic, there was no significant difference between the frequencies of T2 locus genotypes and the frequencies of Q-1,T2 the alleles (P > 0.05). There were no significant differences in T2 locus genotypes and clinical indicators of lung function FEV1% predicted and FEV1/FVC in patient group (P > 0.05). Conclusion The ADAM33 gene Q-1 locus may be related to the COPD susceptibility in Xinjiang Kazak, Han.

Objective To investigate the difference of IOL Master and A-ultrasound combined with corneal topography meas-urement in intraocular lens ( IOL) power calculation .Methods A total of 84 eyes of 60 cataract patients received phacoemulsification and intraocular lens implantation surgery were selected in Xiangya Hospital from March 2010 to September 2011.There were 57 eyes of 42 age-related cataract patients and 27 eyes of 18 complicated cataract patients .The patients were divided into IOL Master group and A ultrasound combined with corneal topography measurement group .Before surgery , in IOL Master group , axial length ( AL) and corneal curvature were measured with IOL Master , IOL power was calculated according to the Haigis , SRK/T formula and so on.At the same time the axial length ( AL) and corneal curvature were measured with A ultrasound combined with corneal topography respectively and IOL power was calculated by SRK/Ⅱ formula.In A ultrasound combined with corneal topography measurement group , axial length ( AL) and corneal curvature were measured with A ultrasound combined with corneal topography respectively , IOL power was calculat-ed according to the SRK/Ⅱformula.3 months postoperatively , all the patients were conducted refractive outcome and calculating mean absolute refractive error(MAE).Finally the data were analysed.Results ⑴In IOL Master group, before operation there was no sig-nificant difference in mean axial length between IOL Master and A-ultrasound measurement ( P >0.05 ) , while in the patients with AL>26 mm the axial length was (28.53 ±0.57)mm and (29.42 ±0.64)mm using IOL Master and A-ultrasound measurement respec-tively ( P <0.05).The mean corneal curvature was (42.12 ±0.31)D and (43.09 ±0.27)D using IOL Master and corneal topogra-phy measurement respectively ( P <0.01).The mean IOL power were (17.06 ±0.48)D and (16.37 ±0.56)D in IOL Master group and A ultrasound combined with corneal topography measurement group respectively ( P <0.05 ) .⑵3 months postoperatively , the MAE was (0.07 ±1.05)D and (0.16 ±0.81)D in IOL Master group and A-ultrasound combined with corneal topography measure-ment group( P <0.05) respectively.Conclusions In cataract patients with normal axial length , IOL Master and A-ultrasound com-bined with corneal topography have high consistency for axial length measurement and IOL power calculation .But in cataract patients with high myopia , IOL Master is more accurate for axial length measurement and IOL power calculation compared to A -ultrasound com-bined with corneal topography .

BACKGROUND: Full-thickness skin graft was a base for bum and plastic surgery, while uniform pressure and regional brake were key factors to ensure skin graft survival and avoid from necrosis. Traditionally, package and pressurized fixation were performed after skin transplantation; however, it induced residual dead space and unclear skin graft fixation, as well as suturing scar.OBJECTIVE: To study the effect of vacuum sealing drainage applied to full-thickness skin graft.METHODS: A total of 8 New Zealand rabbits were used to establish full-thickness skin graft models in three regions of bilateral spine. Vacuum sealing drainage, traditional pressurized suture and common wrapping were performed in the three regions, respectively. The skin graft survival was observed, and survival rate was calculated at 14 and 21 days. On the 3rd, 7th, and 14th days, samples were selected from skin graft and stained. The morphology was observed under light microscope and transmission electron microscope. The time to remove the drain vessel was that when the fluid was not increased or the fluid was clear.RESULTS AND CONCLUSION: The survival rate of vacuum sealing drainage group was significantly higher than that in other two group (P < 0.05), while the survival rate of traditional pressurized suture group was significantly higher than that in the common wrapping group (P< 0.05). Morphology examination demonstrated that regional cuticular layer was necrotic in the vacuum sealing drainage group after early skin transplantation, while inflammatory cell infiltration, fibroblast degeneration, and mitochondrial swelling were also observed. At later skin transplantation, fibroblast and basal cell were proliferated, and function of mitochondria and plasmid was active. This suggested that vacuum sealing drainage promoted survival rate of skin graft.

BACKGROUND: Extended high frequency audiometry is able to detect early cochlea damage. So it is useful in monitoring ototoxic, senile and noise induced hearing loss.OBJECTIVE: To study with extended high frequency audiometry the auditory function of people in highland of qinghai province without noise pollution and evaluate the sensitivity of this method in diagnosing early noise induced hearing loss. To provide the epidemiological data for early control of noise induced hearing loss in this area.DESIGN:It is a transverse comparison among normal people in two areas.SETTING: This study was cooperated by the Otolaryngeal Department of Gulou Hospital affiliated to Nanjing Medical College, the Otolaryngeal Department of Xijing Hospital of the Fourth Medical College and Auditory Research Center of Giessen University, Germany.PARTICIPANTS: Totally 300 people lived in highland of qinghai province received hearing test including extended high frequency audiometry. At the same time another 300 people were chosen in Nanjing city as control group.METHODS: The subjects in the two groups received the whole set of hearing tests including extended high frequency audiometry and answered a questionnaire about their personal history. The mean hearing thresholds from 0. 25-16 kHz between the two groups were compared with t-test.MAIN OUTCOME MEASURES: The difference of mean hearing thresholds from 0. 25-16 kHz between the two groups.RESULTS: There was no significant difference between hearing threshold of low frequencies and speech frequencies. The hearing threshold of high frequencies rose with the stepping-up of testing frequencies which made the possibility of measuring the hearing threshold reduced. But the hearing threshold above 6 kHz of the people in area without noise pollution was significantly higher than that of the control( P ＜ 0.05) . No difference between children and adolescence was found in the 2 groups. The epidemiological data revealed that the people without noise pollution had unperceptible hearing damage compared with the control group.CONCLUSION: The damage of impulsive noise to hearing began at 20 kHz and developed from 20 kHz to 10 kHz. If it was not protected, it would be extended to the speech frequencies and the hearing loss would get worse. So the extended high frequency audiometry was thought helpful to screen noise induced hearing loss early.

ve To understand the levels of trace elements in hair of patients with hypertension disease and coronary heart disease. Methods The contents of zinc, copper, manganese and iron in hair were determined among 45 patients with hypertension disease, 36 patients with coronary heart disease and 40 healthy controls by flame-atomic absorption spectrophotometry respectively. Results The contents of zinc and the ratio of the contents of zinc vs the contents of copper in hair of patient with hypertension disease and coronary heart disease showed significantly higher levels compared with those of healthy controls (P

Objective To determine the content of paeoniflorin with different medicinal parts in Radix Paeoniae Alba and offer a reference for collection and processing. Methods The content of paeoniflorin was measured by HPLC method. Combining air drying with weight relief condition, the content of paeoniflorin was calculated in fresh Radix Paeoniae Alba. Results There was the highest content of paeoniflorin in Radix Paeoniae Alba in 24～48 hours after collection, and the another increasing of the content was founded 120 hours after collection. Conclusion Radix Paeoniae Alba should be processed in 48 hours after collection.