Objective To find out whether NOTCH receptors can serve as direct downstream targets of transforming growth factor β(TGF-β)/Smad4 signaling in endothelial cells.Methods Real-time PCR and Western blotting were performed to verify whether the expression of notch1 and notch4 was regulated by TGF-β pathway.Luciferase reporter assay was employed to investigate how the promoter of notch1 and notch4 was regulated by TGF-β.Then, ChIP assay was used confirm whether the promoter of notch1 and notch4 physically interacted with SMAD protein.Results TGF-β1 and bone morphogenetic protein 4 (BMP4) treatment increased the expression of both notch1 and notch4 at the transcriptional level.In addition, SMAD4 was physically associated with the SMAD binding sites on the notch4 promoter, which was largely enhanced under the treatment of TGF-β1 and BMP4.Importantly, TGF-β1 and BMP4 failed to transactivate notch4 in the absence of endogenous SMAD4 or the SMAD binding regions on the notch4 promoter.Conclusion The expression of NOTCH receptor can be directly up-regulated by SMAD4-mediated TGF-β/BMP signaling in cerebrovascular endothelial cells.

Objective To analyze the distribution and antimicrobial resistance of pathogens isolated from department of hematology during the past three years.Methods Pathogenic strains isolated from patients hospitalized in a hematology de-partment between January 2011 and December 2013 were collected,antimicrobial susceptibility testing was performed by Kirby-Bauer disk diffusion method or automatic system,antimicrobial susceptibility testing results were judged according to American Clinical and Laboratory Standards Institute 2011, data were analyzed by WHONET 5.6 software. Results A total of 462 clinical isolates were collected in 2011—2013,including 161 gram-positive cocci isolates,279 gram-negative bacilli,and 22 fungi.Of Staphylococcus spp ,detection rate of methicillin-resistant coagulase negative Staphylo-coccus(MRCNS)and methicillin-resistant Staphylococcus aureus (MRSA)was 81.37% and 62.50%respectively.The re-sistant rate of Staphylococcus spp .and Enterococcus spp .to linezolid was 1.69% and 3.57% respectively,resistant rate of Staphylococcus spp .to teicoplanin was 3.39%,vancomycin-resistant gram-positive coccus was not found.Enterobacte-riaceae strains Escherichia coli and Klebsiella pneumoniae were highly susceptible to carbapenems,the sensitivity rates were 97.56%—98.88%;while nonfermentative gram-negative bacilli Pseudomonas aeruginosa and Acinetobacter bauman-nii strains were obviously resistant to carbapenems,the resistance rates were 38.71%—64.00%.Conclusion Antimicrobial resistance of major pathogenic strains from hematology department is high,antimicrobial agents should be used according to pathogenic distribution characteristics and antimicrobial susceptibility testing results,healthcare-associated infection control should be strengthened to reduce antimicrobial resistant rate.

Objective To investigate the value of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) in the diagnosis of sarcoidosis.Methods Twenty-two cases of clinically suspected sarcoidosis underwent bronchoscopy and ultrasound bronchoscopy examination,including endobronchial biopsy (EBB),transbronchial lung biopsy (TBLB),and EBUS-TBNA.The biopsy samples of EBB,TBLB,and EBUS-TBNA were paraffin-embedded for hematoxylin eosin (HE) staining and acid-fast staining,respectively.Differences in the diagnosis of sarcoidosis were compared among EBB,TBLB,and EBUS-TBNA.The diagnostic performance of EBUS-TBNA was evaluated.Interventional pulmonology diagnostic strategy of sarcoidosis was analyzed.Results Among all the 22 patients with suspected sarcoidosis,20 cases were diagnosed as sarcoidosis,1 case was small cell lung cancer,and 1 case was lymphoma.The number of patients who were diagnosed by EBB,TBLB,and EBUS-TBNA was 6 cases,9 cases,and 16 cases,respectively;and their diagnostic yield was 30.0％,45.0％,and 80.0％,respectively.The diagnostic yield of EBUS-TBNA was significantly better than the other two (P =0.005).Combined EBB and EBUS-TBNA,the diagnostic yield was 85.0％.Combined TBLB and EBUS-TBNA,the diagnostic yield was 90.0％.Combined those three,the diagnostic yield was 95.0％.EBUS-TBNA diagnostic yield was affected by the location and size of lymph nodes.The diagnostic yield of subcarinal lymph nodes and paratracheal lymph nodes by EBUS-TBNA was significantly better than that of Hilar lymph nodes (x2 =4.29,P ＜0.05),EBUS-TBNA showed better diagnostic yield for the lymph nodes whose diameter was greater than 2cm (x2 =4.067,P ＜ 0.05).EBUS-TBNA had fewer complications.Most patients only had a little bleeding in puncture site.Conclusions EBUS-TBNA contributed to diagnose sarcoidosis,while TBLB and EBB had a complementary value in the diagnosis of sarcoidosis by EBUS-TBNA.

Objective To understand the cognition about healthcare-associated infection(HAI) management among directors in secondary and above hospitals in Shaanxi Province.Methods Questionnaire survey was adopted to investigate hospital directors who participated in The third session of Shaanxi Provincial HAI management training course for hospital directors.Results A total of 181 questionnaires were distributed, 173 (95.58％) were qualified.74.57％ of surveyed hospitals were secondary hospitals, 61.85％ were comprehensive hospitals, 67.05％ of respondents received HAI training in recent 3 years, 81.50％ and 55.49％ of hospital directors thought the main factors influencing the HAI management were health care workers'' awareness on HAI and leaders'' attention respectively.58.96％, 60.12％, and 46.82％ of hospital directors thought the director of HAI management department should have intermediate and above professional title, bachelor degree or above education, and preventive medicine professional requirements respectively.The awareness rate of HAI management-related knowledge was 86.71％, difference in awareness rate of HAI management-related knowledge among respondents of different job, gender, and HAI training in recent 3 years were all significantly different(all P<0.05).Conclusion Hospital directors'' cognition on HAI management affect the development of HAI work, strengthen the training on HAI knowledge among administrators can improve hospital administrators'' awareness on HAI prevention and control.

Objective Stathmin, a microtubule-destabilizing protein , has high expression in esophageal squamous cell carci-noma(ESCC), while taxol is a common chemotherapy microtubule-targeted drug for esophageal cancer .This study aimed to investigate the impact of stathmin expression and its influence on taxol sensitivity in ESCC . Methods We established 2 cell models with ST-MN1 gene overexpression in KYSE 510 and KYSE 170 cell lines, including KYSE 510-Stathmin, KYSE 170-Stathmin, KYSE 510-Control and KYSE 170-Control.MTT assay and colony formation were applied to compare the taxol sensitivity between experimental group and control group .Flow cytometry was used to measure the apoptosis of KYSE 510-Stathmin and KYSE 510-Control after taxol treatment.Western blot was used to test the changes of related factors to apoptosis and autophagy . Results ①Stathmin protein ex-pressions in KYSE 510-Stathmin and KYSE 170-Stathmin cells were higher than those of control cells (P<0.01).② The percentages of inhibition were significantly decreased in KYSE 510-Stathmin and KYSE 170-Stathmin cells 24 h after 50, 100,250 nmol/L taxol treat-ment compared with KYSE 510-Stathmin cells(P <0.01).③The percentages of inhibition were significantly reduced in KYSE 510-Stathmin and KYSE 170-Stathmin cells after 250 nM taxol treatment for 24, 48, 60 h (P<0.01).④After taxol treatment,the number of colony formation in KYSE 510-Stathmin cells was higher com-pared with KYSE 510-Control cells (P<0.01).⑤The percentage of cell apoptosis in KYSE 510-Stathmin was significantly lower than that of KYSE 510-Control cells by flow cytometry (11.90%±0.78%vs 29.63%±3.26%, P<0.05).Western blot showed the ap-optosis of associated proteins such as the activation of Caspase 8 and Caspas9. Conclusion The result indicates that overexpression of stathmin inhibits taxol sensitivity in ESCC cell lines .

Objective:To explore the mechanisms of regulation of miR-575 on the proliferation and invasion properties of non-small cell lung cancer cell (NSCLC).Methods:Real-time PCR was selected to detect the expression of miR-575 and BLID in differ ent NSCLC cell lines.CCK-8 assay was processed to measure the alternations of A549 cell proliferation at different time points after transfection of miR-575 mimic and miR-575 inhibitor.The invasion ability of A549 cells was evaluated by transwell.The targeting of BLID by miR-575 was predicted by Targetcan software and verified by dual-Luciferase assay.BLID protein expression level was detected by western blot.Results:miR-575 highly expressed in NSCLC cell lines,including A549,SPC-A1,H1299,H1650 (P＜0.001),miR-575 mimic could efficiently elevated the expression ofmiR-575 in A549 cells (P＜0.001),and strengthened the proliferation and invasion ability of NSCLC cells (P＜0.05),while,transfection of miR-575 inhibitor could down-regulate the expression ofmiR-575,and also inhibit the proliferation and invasion ability of NSCLC cells (P＜0.01).Targetscan software predicted that BLID might be the target gene of miR-575,and dual-luciferase assay revealed that miR-575 could obviously decrease the luciferase reaction of wild type BLID 3'UTR (P＜0.01),besides,miR-575 could down-regulate the protein expression ofBLID (P＜0.01).Real-time PCR results showed that NSCLC cell lines had lower level of BLID mRNA expression compared with 16HBE control cells (P＜0.001),and restore of BLID could markedly inhibited cell proliferation and invasion ability (P＜0.05),which could be reversed by miR-575 co-tranfection (P＜0.01).Conclusion:In NSCLC cells,the expression ofmiR-575 could promote cell proliferation and invasion ability by directly regulating downstream target tumor-suppressor gene BLID expression.

Objective: To investigate the feasibility of in vitro perfusion culture of human adipose tissue and its induction into muscle tissue. Methods: Human abdominal adipose tissue were cultured in vitro by perfusion culture. After 1, 3, 5 or 7 weeks, FAD/PI staining was used to detect the tissue vitality. Histological staining was used to observe the changes of its histomorphology. Protein expressions of myogenic molecules Myf-5 and myoD1 as well as muscle specific protein Desmin were measured by immunohistochemistry and Western blot assay. Results: The adipose tissues cultured in myogenic induction media were still in the appearance of adipose tissue at 7 weeks. While in the basal medium without inducing, vascular pedicles shed after 7 weeks and could not continue to be cultured. FAD/PI staining showed that the tissue cultured in the induction media remained viable at 7 weeks, while the viability of the tissue in the basal culture medium decreased significantly at 5 weeks. Histologically, Myf-5, myoD1 and Desmin were all positively expressed in muscle tissues, while in adipose tissues, some mesenchymal and vascular endothelial cells expressed Myf-5 but not myoD1, and only separate vascular smooth muscle cells expressed Desmin. Interestingly, in adipose tissues cultured in myogenic induction medium, partial muscle-like tissue formed, evidenced as positive expression of Myf-5, myoD1 and Desmin. There was no muscle-like tissue formation in adipose tissue cultured in basal medium and the expression patterns were similar to that of the control group. Western blot results showed that the expression levels of Myf-5 and Desmin in muscle tissue were significantly higher than that of the other groups (P<0.05). The expression level of Myf-5 was slightly higher in inducing group than that in non-inducing group at the same time points, but the difference was not statistically significant. Similarly, the expression of Desmin was higher in inducing group than that in non-inducing group, yet there was statistically significant difference only in the first week. The protein expression of myoD1 was not detected in any group using Western blot. Conclusions The adipose tissue cultured in the perfusion bioreactor can survive for up to 7 weeks, and it can be partially induced into muscle-like tissue in the myogenic induction medium.

MicroRNA (miRNA) is a type of small non-coding RNA and acts as a post-transcriptional regulator of gene expression. This article briefly describes the etiology of various chronic liver diseases, including metabolic dysfunction-associated fatty liver disease, chronic hepatitis B, chronic hepatitis C, chronic drug-induced liver injury, liver cirrhosis, and hepatocellular carcinoma, and summarizes related reports on microRNA-125b which enters different signal transduction pathways and plays the same or contradictory regulatory role in the same liver disease or pathological process by targeting different target genes, so as to provide insights into the research on the pathogenesis of various chronic liver diseases and the establishment of non-invasive differential methods.

The column chromatography on silica gel, sephadex LH-20 preparative HPLC were used to separate and purify the compounds from the stems of Dendrobium candidum. Twenty compounds were isolated and identified as 3,4'-dihydroxy-5-methoxybibenzyl(1), dihydroresveratrol(2), dendromoniliside E(3), denbinobin(4),2,4,7-trihydroxy-9, 10-dihydrophenanthrene(5), aduncin(6), (-)-loliolide(7), adenosine(8), uridine(9), guanosine(10), sucrose(11), 5-hydroxymethyl-furaldehyde(12), n-octacostyl ferulate(13), defuscin(14), n-triacontyl cis-p-coumarate(15), daucosterol(16), beta-sitosterol(17), hexadecanoic acid(18), hentriacontane(19), and heptadecanoic acid(20). Their structures were elucidated on the basis of spectroscopic data and physicochemical properties. All of the compounds were isolated from this plant for the first time.
Dendrobium ; chemistry ; Molecular Structure ; Plant Extracts ; chemistry ; isolation & purification

Objective: To understand the effect of sirolimus on the erythropoiesis of K562 cell line and bone marrow cells from pure red cell aplasia (PRCA) patients and normal controls. Methods: Different concentrations (10, 100, 1 000 nmol/L) of sirolimus were added to the K562 cell line or bone marrow cells from PRCA patients or normal controls and cultured 14 days for BFU-E formation. Meanwhile, sirolimus was also added to the serum treated PRCA bone marrow cells to cultivate for the same priod of time. Results: Neither K562 cells, bone marrow cells from PRCA patients or normal controls showed any difference when sirolimus was added to the culture system for BFU-E. However, BFU-E formation decreased after serum was added in PRCA patients (76.40±22.48 vs 136.33±12.58, t=-4.329, P=0.001) and this suppression of BFU-E was partly corrected by 1 000 nmol/L sirolimus treatment (97.14±15.83 vs 76.40±22.48, P=0.038). Conclusions Sirolimus may modulate the suppression of erythropoiesis by serum instead of directly stimulate the growth of red blood cells in PRCA patients.