Objective To study the association of the expression of bone morphogenetic protein (BMP) antagonist gremlin and vascular calcification in radial arteries of patients with stage 5 of chronic kidney disease (CKD).Methods Radial arteries of 40 patients with stage 5 of CKD were collected as specimens of the study group,which were trimmed off during arterial venous fistula operations.Splenic trabecular arteries were collected as specimens of the control group,which were removed from 38 patients with simple traumatic splenic rupture.All the arteries were examined histologically for calcification with yon Kossa stain.Expressions of gremlin and BMP-2,-7were detected by immunohistochemistry and their serum concentrations were detected by ELISA.Images of histological sections were semi-quantitatively analyzed by Image-Pro Plus 6.0.SPSS 19.0software was used to perform statistical analysis.Results Significantly positive von Kossa stain was found in radial arteries from 12 of 40 patients (30％) in study group,which located in the layer of medial smooth muscle cells.However,there was no obvious positive stain in control group.Additionally,in study group,significant expressions of gremlin and BMP-2 were detected in those radial arteries of positive yon Kossa stain,which also located in the layer of medial smooth muscle cells.Positive correlations were found among gremlin expression level,BMP-2 expression level and yon Kossa stain intensity.However,the BMP-7 expression intensity in arteries of study group was much weaker as compared to control group.Conclusions Both gremlin and BMP-2 may be involved in the process that the smooth muscle cells of radial arteries in patients with stage 5 of CKD phenotypically transform into osteoblast-like cells.However,BMP-7 possibly prevents this process.

Objective To construct a cDNA expression library from unfed female tick Haemaphysalis longicornis for screening and cloning potential antigenic genes.Methods Total RNA was isolated from unfed female ticks,mRNA was purified and a library of oligo(dT)-primed cDNA with added directional EcoR Ⅰ/Hind Ⅲ linkers was constructed from the purified mRNA.The constructed cDNA was ligated to the EcoRⅠ/HindⅢ arms of the ?SCREEN vector.Pure phage stocks were harvested by plaque purification and converted to plasmid subclones by plating phage on host strain BM25.8.Recombinant plasmids that were subcloned to E.coli BM25.8 were isolated and transformed into E.coli JM109.Recombinant plasmids abstracted from JM109 were identified by PCR and sequencing.Rusults The recombinant phage DNA was packaged by using phage-marker packaging extracts,resulting in a primary cDNA library with a size of 1.8?106 pfu.Data showed 100% of the library were recombinant and the titer of the amplified library was 2.4?109 pfu/ml.Forty-two clones of encoding immunodominant antigens were obtained from the cDNA library.Sequence analysis revealed 12 unique cDNA sequences and the encoded putative proteins showed similarities to H.longicornis tropomyosin mRNA,Rhipicephalus annulatus unknown larval protein mRNA,chromosome 2R of Drosophila melanogaster,mitochondrial DNA of H.flava,clones HqL09 unkown mRNA and Hq05 mRNA of H.qinghaiensis,and myosin alkali light chain protein mRNA.Conclusion The cDNA expression library from unfed female H.longicornis was successfully constructed and screening of protective genes may provide candidate antigens of the tick.

Objective To study the activation of alveolar macrophage β (AM) and the expression of co-stimulatory molecule CD40 in transfusion-related acute lung injury (TRALI) model in order to illustrate the pathogenesis of TRALI.Methods Sixty SD rats were randomly (random number) divided into normal control group (n =15) with sham operation using normal saline instead of LPS and plasma,positive control group (n =15) with ALI induced by intravenous infusion of 5 mg/kg lipopolysaccharide (LPS) in equivalent volume of whole blood drawn out),and TRALI group (n =15) treated by intra-peritoneal injection of 2 mg/kg LPS 2 h before the transfusion of human plasma (1 mL whole blood about 10％ of total blood volume drawn out and replaced with 1 mL plasma),LPS control group (n =15) treated by intra-peritoneal injection of 2 mg/kg LPS 2 h before saline infusion in equivalent volume of blood drawn out.The pathologic changes of rat lung tissue were observed by HE staining.The expression of TLR4 was examined by RT-PCR.The activation of NF-κB in AM was measured by electrophoresis mobility shift assay (EMSA).The expression of CD40 mRNA and CD40 molecule were analyzed by Northern blot and flow cytometry respectively.ELISA was performed to detect the concentration of TNF-α,MIP-2 and IL-1 β in broncho-alveolar lavage fluid (BALF).Results Broken alveolar septa,hyperemia,and massive infiltration of inflammatory cells including the neutrophils were observed in lung tissues of TRALI group.The expression of TLR4 gene was detected in activated macrophage phi (AMφ) of TRALI group rats.The activation of NF-κB was increased in TRALI group rats.The expression of CD40 in AMφ was higher in rats of TRALI group than that in rats of control group and LPS control group.The concentration of TNF-αt,MIP-2 and IL-1β were enhanced significantly in BALF of TRALI group rats.Conclusion The activation of AM and up-regulation of costimulatory molecule CD40 induced release of some inflammatory cytokines.It suggested that AM activation may play an important role in the pathogenesis of TRALI.

Objective To evaluate regulatory effects of miRNA-146a on peripheral blood CD4 + T lymphocytes from patients with psoriasis vulgaris, and to investigate the role of miRNA-146a in the pathogenesis of psoriasis vulgaris. Methods Totally, 30 patients with psoriasis vulgaris and 30 healthy human controls were enrolled into this study. Venous blood samples were obtained from these subjects, and CD4 + T lymphocytes were isolated from these samples by using magnetic activated cell sorting (MACS). Real-time quantitative PCR (RT-PCR)was performed to measure the expression of miRNA-146a in peripheral blood CD4+ T lymphocytes, and enzyme-linked immunosorbent assay(ELISA)to determine plasma levels of interferon-γ(IFN-γ)and interleukin 4(IL-4). Some CD4+ T lymphocytes were divided into 3 groups to be transfected with 50 nmol/L negative control miRNA (control group), miRNA-146a mimics(miRNA-146a group)or miRNA-146a inhibitor (miRNA-146a inhibitor group). After 24-hour additional culture, flow cytometry was conducted to determine the number of Th1 and Th2 cells, Western-blot analysis and RT-PCR were performed to measure the protein and mRNA expressions of IFN-γ receptor α (IFN-γRα), T-box expressed in T cells (T-bet)and GATA-binding protein-3 (GATA-3)respectively, and ELISA was carried out to determine the levels of IFN-γ and IL-4 in supernatants of CD4 + T lymphocytes. Results Compared with the healthy control group, the patient group showed significantly increased miRNA-146a expression in peripheral blood CD4 + T lymphocytes (243.81% ± 94.32% vs. 105.74% ± 22.93%, t = 6.653, P < 0.01)and plasma IFN-γ level (27.69 ± 7.64 ng/L vs. 9.75 ± 2.81 ng/L, t = 4.237, P <0.01). Moreover, miRNA-146a expression was positively correlated with plasma IFN-γ level in the patients(r = 0.837, P <0.01). After 24-hour culture in vitro, there was a significant increase in the number of Th1 cells, protein and mRNA expressions of T-bet, and supernatant level of IFN-γ, but a significant decrease in the protein expression of IFN-γRα in the miRNA-146a group compared with the control group (all P < 0.01). However, no significant differences were observed in the number of Th2 cells, mRNA or protein expressions of GATA-3, or supernatant level of IL-4 among the control group,miRNA-146a group and miRNA-146a inhibitor group (all P > 0.05). Conclusion miRNA-146a may play an important role in the pathogenesis of psoriasis vulgaris by participating in the regulation of peripheral blood CD4+ T lymphocytes via affecting Th1 cell differentiation and function.

Objective:To study effects of large dose thymopeptides on T cell subpopulations of patients with malignant tumor under radiotherapy.Methods:Fifty one patients with malignant tumor under radiotherapy were divided into 2 groups with 100 mg and 200 mg thymopeptides respectively.The patients we re given thymopeptides,100 mg/d or 200 mg/d,iv, for 10 days.The positive percent ages of CD4,CD8,CD25 and CD56 in T cells of peripheral blood before and after thymopeptide treatment were determined by flow cytometry.Results:The positive percentages of CD4 and CD25 in T cells of peripheral bl ood after 100 mg/d thymopeptide treatment were significantly higher than those befor e thymopeptide treatment (P<0.05),while those of CD4，CD8，CD25 and CD56 in T cells of peripheral blood after 200 mg/d thymopeptide treatment all increased significantly (P<0.05 or P<0.01). Conclusion:These results suggest that large dose of thymopeptides can increase i mmune function of patients with malignant tumor under radiotherapy,and the curat ive effect of 200 mg/d thymopeptides is better.

Objective To investigate the value of early?phase enhancement ratio combined with peripheral vascular diameter in the differential diagnosis of benign and malignant breast lesions using 3.0 T dynamic contrast?enhanced magnetic resonance imaging (DCE?MRI). Methods Sixty seven cases of patients (35 with malignant lesions and 32 with benign lesions in the breasts) were retrospectively analyzed. Their diagnoses were confirmed by surgery and pathology and all the patients underwent breast MRI plain scan and DCE?MRI in the two weeks before surgery. Lesion ROIs were drawn and time?signal intensity curves in the DCE?MRI were generated. Early?phase enhancement rate, time to peak, early?phase enhancement ratio, numbers of tumor vessel within 3 cm of the lesion and diameter of the largest vessel were recorded. Mann?Whitney U test was used to compare the difference of DCE?MRI between benign and malignant lesions, and the ROC curve was used to evaluate the efficiency of early?phase enhancement rate, early?phase enhancement ratio and vascular diameter in differentiating benign and malignant lesions. Results With breast malignant lesions, the medians of time to peak, early?phase enhancement rate, early?phase enhancement ratio, numbers of tumor vessel and vascular diameter were 2.2 s, 176.0%, 100.0%, 4 and 2.96 mm respectively, while with benign lesions of these parameters were 4.7 s, 113.3%, 81.9%, 0 and 0.00 mm respectively, and the differences were statistically significant (all P<0.05).When early?phase enhancement rate was used for differential diagnosis of breast benign and malignant lesions, the area under the ROC curve was 0.702 and the sensitivity and specificity were 82.86%and 56.25%with a threshold of 120.0%. When early?phase enhancement ratio was used, the area under the ROC curve was 0.854 and the sensitivity and specificity were 94.29% and 68.75% with a threshold of 86.0%. When peripheral vascular diameter was used, the area under the ROC curve was 0.896 and the sensitivity and specificity were 74.29%and 84.38% with a threshold of 2.78 mm. When early?phase enhancement ratio was combined with peripheral vascular diameter, the area was 0.925 and the sensitivity and specificity were 97.14% and 62.50%. Conclusion In the differential diagnosis of benign and malignant breast lesions under DCE?MRI, early?phase enhancement ratio combining with peripheral vascular diameter has improved sensitivity.

Objective: To investigate anxiety and associated factors among college students returned to school during COVID-19 epidemic, and to provide reference for anxiety relief and school life adaptation to help college students. Methods: A cross-sectional study was conducted among 4 750 students selected from Nanjing Medical University, China Pharmaceutical University, Nanjing University of Chinese Medicine from 2 May to 9 May, 2020. Zung Anxiety Self-rating Scale (SAS) was used to measure anxiety status of students. The self-made questionnaire was used to investigate the related factors. Results: During COVID-19 epidemic situation ,20.0% college students returned to school had mild anxiety, 6.5% had moderate or severe anxiety. College students unable to adapt to campus closed-off management with a higher anxiety detection rate of 52.9%.Students with heavy study burden whose offline study time per day ≥6 hours have a higher anxiety detection rate of 36.0%. Students never exercising had a higher anxiety detection rate of 43.5%. The detection rate of anxiety not participating in online sports and sports activites was 41.6%. 40.7% students were detected of anxiey because of loneliness caused by not being able to meet friends. Multiple logistic regression analysis revealed that the adaptation to the campus closed-off management, satisfaction with the campus disinfection, participation in group activities, the time of offline study per day, the time of online study per day, tension about the exams, regular routine, exercise, participation in online activities were the influencing factors related to anxiety. Conclusion During COVID-19 epidemic some of college students exhibit anxiety when back to school life, which might be associated with campus management, study burden, exercise and regular routine, school activities and so on. Schools should take measures to relieve students anxiety while epidemic prevention.

ObjectiveTo investigate the diagnostic value of EUS-FNA for pancreatic masses and correlated influential factors. MethodsWe retrospectively analyzed the clinical data of 101 patients with pancreatic lesions who underwent EUS-FNA from January 2008 to January 2010. The clinical data enrolled 10 factors including patient gender, patient age, lesion location, lesion size, lesion characteristics, negative suction pressure, times of access, real-time cytological diagnosis, type of EUS and operators' experiences.ResultsThe overall diagnostic accuracy, sensitivity, specificity, positive predictive value and negative predictive value of EUS-FNA were 85. 1％, 81.1％, 96. 3％, 98. 4％ and 65.0％, respectively. Univariable logistic regression analysis indicated that lesion size, lesion characteristics, negative suction pressure, operators' experience were correlated factors of EUS-FNA positive rate, while lesion size was the only correlated factor of EUS-FNA diagnostic accuracy ( OR =1. 984,95 ％ CI: 1. 141 ～ 3. 451, P =0. 015 ). Every 1 cm the lesion increased, by 1.67 times of opportunity the positive rate became, by 1.83 times of opportunity the accuracy was. The lesion size and lesion characteristics were independent correlated factors of EUS-FNA positive rate (OR=2.012, P=0.000; OR =10.218, P=0. 002). The positive rate of EUS-FNA in solid lesions was 10. 2 times of that in cystic lesions. Lesion size was the independent correlated factors of EUS-FNA diagnostic accuracy (OR =1. 984, P =0. 015 ). ConclusionEUS-FNA can effectively make a pathological diagnosis of pancreatic masses with high diagnostic accuracy and specificity. EUS-FNA diagnostic positive rate and accuracy were both positively correlated with pancreatic lesion size. EUS-FNA positive rate of solid pancreatic lesions is significantly higher than that of cystic lesions.

Objective To provide an objective basis for differential diagnosis of pancreatic diseases through quantitative analysis of the different features of contrast-enhanced endoscopic ultrasonography (CE-EUS). Methods A total of 32 patients with suspected or confirmed pancreatic neoplasms or chronic pancreatitis and 19 patients who underwent EUS due to other digestive problems other than pancreatic disease were enrolled. Features of blood perfusion of the regions of interest during CE-EUS were analyzed quantitatively. The findings were compared with cytological and/or histopathological results of EUS-FNA and/or surgery.Results Quantitative analysis of CE-EUS showed peak intensity (PI) value of 19 normal pancreas was 0.648 ±0. 174, which was statistically different from that of pancreatic cancer and pancreatic cystic lesions. Based on ROC, the cutoff of differential diagnosis was 0. 505, and the sensitivity and specificity were 100. 0% and 84. 2%, respectively. PI value of 6 chronic pancreatitis was the highest (0. 772 ±0. 106). In pancreatic neoplams, PI values of pancreatic carcinoma, pancreatic cyst and pancreatic endocrine tumors were significantly different. Based on a cutoff of 0. 195, the sensitivity and specificity of differentiation of pancreatic carcinoma and pancreatic cyst were 85.7% and 87.5%, respectively. PI value of 14 pancreatic carcinoma and that of 4 pancreatic endocrine tumors were 0. 321 ± 0. 119 and 0. 763 ± 0. 115, respectively. Through the comparison between the AT and TTP of the focal lesions and surrounding pancreatic parenchyma, 78.6% pancreatic carcinoma showed slow falling-in and rapid wash-out and all the endocrine tumors showed rapid falling-in and rapid wash-out. The PI value of 8 patients with pancreatic cyst was 0. 181 ±0. 036, with no enhanced blood flow in the cyst. The TIC was a straight line. Conclusion CE-EUS with quantitative analysis is a promising method that can be a more objective basis in the differential diagnosis of pancreatic diseases.

Objective To evaluate the impact of breathing motion on target volume and the factors influencing the set-up errors during tangential whole breast irradiation.Methods From Jan 2003 to Dec 2003,patients with early-stage breast cancer after breast conserving surgery,were selected to be eligible for the study.All patients were immobilized in treatment position by breast beard of Med-Tec 250.The motion of the breast treatment volume was observed on a fluoroscope in different directions under free breathing in 16 patients.The set-up errors in different dimensions during irradiation were measured by weekly portal films (PF) in comparison with digital reconstructed radiographs (DRR) in 11 patients.Results The central lung distance (CLD) variation during free breathing was (2.1?1.2) mm which is greater than the motion to- wards the other directions.By comparing the PF and DRR,the systemic error,random error and overall er- ror in the outer,inner and cranio-caudal directions was 1.9,1.6,2.5 and 2.4,1.7,3.1 and 2.6,2.3, 3.5 mm,respectively.In addition,the discrepancy of the treatment position in cranio-caudal direction and breast volume was most obvious at the beginning 2 weeks with the peak of breast volume at the second week. It decreased gradually during the following 3 weeks.Conclusions This study suggests that the mean value of the motion of the breast target volume during one breathing cycle is less than 2 mm.The set-up errors dur- ing irradiation is the greatest in cranio-caudal direction,suggesting that the fixing precision of the breast board should be further improved.The set-up error during irradiation are most obvious at the beginning two weeks,with the peak of the breast volume in the second week.