Objective To measure and compare the expression profdes of plasma miRNA between Tibetan and Han nationality.Methods Plasma samples were taken from 246 healthy Tibetans and 128 Han individuals.The randomly selected 50 Tibetan and Han plasma samples were pooled respectively and the levels of 754 miRNAs were examined using a TaqMan Low Density Array.Two markedly differentially expressed miRNAs,miR-130a-3p and miR-629-5p,were verified in all the plasma samples by individual qRT-PCR.Results The Low Density Array results showed that the correlation coefficient of expression profiles of plasma miRNA for the Tibetan and Han population was 0.592.Compared with Han population,the expression levels of 139 miRNAs were distinctly different,in Tibetan (62 up-regulated and 77 down-regulated).The levels of miR-130a-3p and miR-629-5p were further verified to be significantly higher in the plasma from Tibetans than those in the plasma from Han population [(467 ± 27.30) × 10-5 vs (236 ± 9.69) × 10-5,p ＜ 0.01;(14.67 ±0.94) × 10-5 vs(7.58 ± 0.52) × 10-5,P ＜ 0.01] by qRT-PCR assay.Conclusion There may be marked difference of plasma miRNA expression profile between Tibetan and Han nationality.The influence of the nationality factors on miRNA profiles should be taken into account in the application of miRNAs in clinical detection in the future.

Objective To explore the safety and effectiveness of diagnostic ultrasound associated with microbubbles to open the blood brain barrier(BBB).Methods Microbubbles were injected through caudal vein,the rat head was radiated by GE Vivid 7 diagnostic ultrasound immediately.The radiated depth was located in the basal ganglia assisted by magnetic resonance imaging(MRI)scanning.The degree of BBB opening was evaluated by enhanced MRI and Evans blue dyeing.The safety was inspected by observation of cell morphology under hematoxylin eosin(HE)staining.Results After the rat head radiated by diagnostic ultrasound with microbubbles,signal enhancement of the radiated area was observed on post contrast T1-weighted images.Red fluorescence of Evans blue was detected by fluorescence microscope in the same area.Normal cellular morphology and structural integrity were showed by HE staining.Conclusions The BBB of rat could be opened targetedly and noninvasively by diagnostic ultrasound associated with microbubbles.This may provide a new strategy for the drugs and stem cells treatment in the central nervous system diseases.

Proteins encoded by regions of difference (RD) of Mycobacterium tuberculosis (MTB) constitute a potential source of specific antigens for vaccine development and immunodiagnosis .In the present study ,four genes named nrdF1 , pe_pgrs35 ,rv1985c ,and rv1986 from RD2 of MTB were cloned and overexpressed in E .coli with the induction of IPTG .Western blotting assay showed that these recombinant fusion proteins could well react with anti-His tag monoclonal antibody ,which in-dicated their good immunoreactivity .The serodiagnosis potential applications of these four proteins in bovine tuberculosis were further evaluated .An indirect ELISA assay was established by using fusion proteins as coating antigens for detection of their specific antibodies in bovine sera .The positive rates were 7 .35% in NrdF1 ,22 .06% in PE_PGRS35 ,16 .18% in Rv1985c and 16 .18% in Rv1986 respectively .All the results suggest that these fusion proteins have the potential application in serodiagnosis of bovine tuberculosis .

Objective To explore the significance of combined detection of D-Dimer,cardic troponin I(cTnI)and N-terminal pro-brain natriuretic peptide(NT-ProBNP)in acute coronary syndrome and the correlation between them.Methods 143 patients with acute coronary syndrome were selected as the observation group,and 40 CAG negative people as the control group.The difference between the two groups was compared and the correlation was analyzed.According to different diagnostics,patients in the observa-tion group were separated into 3 groups,unstable angina pectoris,non ST segment elevation myocardial infarction and ST segment elevation myocardial infarction.The correlation of D-Dimer,cTnI and NT-ProBNP with the severity of coronary artery disease was analyzed.Results The levels of D-Dimer,cTnI and NT-ProBNP in the observation group were higher than those in the control group,and the difference was statistically significant(P < 0.05 ).The severity of coronary artery disease had positive correlation with the test results (P <0.05).Conclusion The combined detection of D-Dimer,cTnI and NT-ProBNP could help to diagnose the acute coronary syndrome and the test result has a positive correlation with the severity of acute coronary syndrome.

Cell therapy has achieved tremendous success in regenerative medicine in the past several decades. However, challenges such as cell loss, death and immune-rejection after transplantation still persist. Biomaterials have been designed as carriers to deliver cells to desirable region for local tissue regeneration; as barriers to protect transplanted cells from host immune attack; or as reactors to stimulate host cell recruitment, homing and differentiation. With the assistance of biomaterials, improvement in treatment efficiency has been demonstrated in numerous animal models of degenerative diseases compared with routine free cell-based therapy. Emerging clinical applications of biomaterial assisted cell therapies further highlight their great promise in regenerative therapy and even cure for complex diseases, which have been failed to realize by conventional therapeutic approaches.
Animals ; Biocompatible Materials ; chemistry ; pharmacology ; Bioreactors ; Cell- and Tissue-Based Therapy ; adverse effects ; methods ; Drug Carriers ; chemistry ; pharmacology ; Humans ; Regenerative Medicine ; methods

No abstract available.
Computational Biology ; Recurrence ; Urinary Bladder Neoplasms ; Urinary Bladder

Objective To evaluate the impact of clinical and pathological factors on incidence and prognosis of brain metastasis from breast cancer.Methods The clinical data of 4473 stage Ⅳ breast cancer patients were collected from The Surveillance,Epidemiology,and End Results (SEER) database.All these patients were divided into brain metastasis group (n=345) and non-brain metastasis group (n=4 128).The clinical and pathological factors between the two groups were compared using x2 test and binary Logistic regression.The OS of 345 brain metastasis patients was calculated using the Kaplan-Meier method and impact of clinical and pathological factors on the survival rate was analyzed by Log-rank method.Multivariate Cox regression model was used to find out the independent risk factors.Results Between brain metastasis group and non-brain metastasis group,these following clinical and pathological factors showed the scatistical significance on brain metastasis:age,histological grade,T stage,N stage,ER,PR and HER expression (P＜0.05).Binary Logistic regression results showed that age,T stage,N stage,ER,PR were independent factors affecting brain metastasis (P＜0.05).The prognosis analysis showed that marital status,histological grade,ER,PR,molecular subtype,radiotherapy and chemotherapy,surgery at primary lesions,surgery at metastasis site,liver metastasis,lung metastasis and numbers of other organs metastasis were significantly correlated with the survival of breast cancer patients with brain metastasis (P＜0.05).Cox regression analysis showed that histological grade,ER,PR,molecular subtype,radiotherapy and chemotherapy,surgery at primary lesions,surgery at metastasis site were independent prognostic factors in breast cancer patients with liver metastasis.Conclusions Breast cancer patients with brain metastasis have a poor prognosis.Molecular subtype should be considered for prediction of prognosis for patients with brain metastasis.Radiotherapy and surgery at metastasis or primary site may improve survival of patients.

Objective To observe the expression of cysteine-rich protein 61 (Cyr61) in transforming growth factor-β1 (TGF-β1)-activated renal fibroblasts (NRK-49F),and to explore its effect and mechanism.Methods (1) NRK-49F cells were activated by TGF-β1 with different concentrations (0.0,0.5,1.0,2.0,5.0 μg/L).Western blotting was used to detect the expression of Cyr61 protein,and CCK-8 assay was used to test the proliferative activity of NRK-49F cells.(2) NRK-49F cells with low expression and over expression of Cyr61 were established by plasmid transfection.The cells were divided into control group (null vector transfection),over-expression group and lowexpression group.The proliferation was discovered by CCK-8 assay after 24,48 and 72 h.Further,5.0 μg/L TGF-β1 activated these three groups.The proliferation was also discovered by CCK-8 assay and the cell cycle was analyzed by flow cytometry.The mRNA expressions of fibrosis markers (Collα1,Col3αl,MMP9,MMP13) and factors of cell senescence signal pathway (p53,p21,Rb,p16) were ascertained by real time PCR,and the protein expressions of Col3 and MMP9 were detected by Western blotting.Results (1) Compared with 0.0 μg/L TGF-β1 group,the proliferation of NRK-49F cells was enhanced in 0.5,1.0,2.0 and 5.0 μg/L TGF-β1 groups (all P ＜ 0.05),while the expression of Cyr61 protein was decreased in 1.0 μg/L group and increased in 5.0 μg/L group (all P ＜0.05).(2) The proliferation of over-expression group was lower than that of control group after 24,48and 72 h (all P＜ 0.05),which was in a time-dependent manner.(3) Compared with control group activated by TGF-β1,the over-expression group expressed less fibrosis factors (Col1α1 and Col3α1)and more anti-fibrosis factors (MMP9 and MMP13) with decreased proliferation (all P ＜ 0.05).Simultaneously,the proportion of cells bogged down in G1 phases,as well as the expressions of p53,p21 and Rb mRNA increased (all P ＜ 0.05).The above effects of low-expression group were just opposite to over-expression group.Moreover,there was no significant difference in the expression of p16 gene among the three groups (P ＞ 0.05).Conclusions Cyr61 can curb the proliferation and fibrotic phenotypes of fibroblasts,thereafter slowing down the process of renal fibrosis.The p53/p21/Rb interrelated cell senescence signal pathway may be involved in the anti-fibrosis process.

Objective To explore the risk factors of post-hepatectomy acute kidney injury (AKI),a Nomogram predictive model of secondary AKI after hepatectomy was established which can provide guidance for the selection of clinically relevant treatment plans and improve the prognosis of surgical patients.Methods A total of 2769 patients who underwent hepatectomy in the Affiliated Hospital of Qingdao University from October 2012 to July 2018 were included in the study.The post-hepatectomy AKI was diagnosed according to the KDIGO AKI criteria in 2012.The selected patients were divided into AKI group (n=133) and non-AKI group (n=2636);they were divided into training group (n=2050) and test group (n=719) according to Enrollment time.The differences of preoperative clinical data,length of hospital stay,and in-hospital mortality between patients with AKI and non-AKI group were compared.The risk factors of post-hepatectomy AKI were evaluated by the Cox regression.A Nomogram predictive model of AKI after hepatectomy was established,and receiver operating curve (ROC) and consistency curve were used to verify the accuracy of the predictive model.Results The incidence of AKI after hepatectomy was 4.80％(133/2769).Compared with non-AKI group,preoperative serum albumin,hemoglobin,and hematocrit levels were lower in AKI group;the level of blood transaminase,total bilirubin,alkaline phosphatase,triglyceride,lactate dehydrogenase,and fibrinogen were higher (P ＜ 0.050);the proportion of preoperative aspirin application was higher (P ＜ 0.001);the duration of operation was longer (P=0.002);the proportion of open surgery was higher (P ＜ 0.001);the mortality rate was higher (P ＜ 0.050);the length of hospital stay was longer (P ＜ 0.050).Cox regression results showed hypertriglyceridemia,hypoalbuminemia,alkaline phosphatase,aspartate aminotransferase,open surgery,lower preoperative glomerular filtration rate,aspirin and duration of surgery were independent risk factors for AKI.We incorporated these indicators into the Nomogram to establish a predictive model for AKI after hepatectomy,the area under ROC curve was 0.764.The area under ROC curve of the test group was 0.781.Conclusion The Nomogram predicting model of AKI after hepatectomy has high accuracy,which is helpful for prognosis of patients who underwent hepatectomy.

Objective To investigate the possible effects of histone methyltransferase MLL1 on renal interstitial fibrosis and epithelial-mesenchymal transition(EMT).Methods Forty-two male SD rats were randomly divided into normal group,sham operation group and unilateral ureteral occlusion(UUO)group,and then UUO group was further divided into group 1 d,1 week,2 week,3 week and 4 week after operation.The expression of MLL1,E-cadherin,α-SMA,Vimentin and Col3α1 in UUO rat kidney tissue as well as TGF-β1 stimulated HK-2 cells were detected by real-time PCR and Western blotting.siRNA-MLL1 plasmids was used to inhibit the expression of MLL1 and the protein levels of MLL1,α-SMA,Vimentin,E-cadherin,Col3α1 and H3K4me3 induced by TGF-β1 stimulation were detected by Western blotting.The level of H3K4me3 in promoter region of EMT related genes was observed by chromatin immunoprecipitation(CHIP).Results Compared with normal and sham operated groups,the loss of renal function in UUO group was more obvious with the obstruction time(P ＜ 0.05).The renal fibrosis was most obvious 1 week and 2 weeks after the rats underwent the UUO operation(all P ＜ 0.05),with the highest protein expressions of MLL1,E-cadherin,α-SMA,Vimentin and Col3α1(all P ＜ 0.05).Compared with the control group,3 ng/ml TGF-β1 induced the highest expression of MLL1 and the most obvious EMT in HK-2 cells(all P ＜ 0.05).Moreover,the EMT and the high level of H3K4me3 in HK-2 triggered by TGF-β1 were all inhibited by siRNA-MLL1 plasmids transfection(all P ＜ 0.05).Conclusions MLL1 can enhance the occurrence of EMT induced by TGF-β1 in HK-2 cells by increasing the level of H3K4me3 in the promoter region of α-SMA and Vimentin.