Objective To study the effects of eukaryotic expressive mature peptide LL-37 of human cationic antimicrobial peptide(hCAP-18) on the expressions of membrane molecules on dendritic cells(DCs).Methods By gene cloning,the eukaryotic expressive plasmid pcDNA4/Myc-His-LL-37 for the mature peptide LL-37 of hCAP-18 was constructed.Then they were transfected into HEK293 cell lines.After the cell lines were cultivated for 48 h,the supernatant was collected.Then the DCs from peripheral blood mononuclear cells(PBMCs) induced by rh-GM-CSF and rh-IL-4 were cultivated with the supernatant for 48h.The expressions of membrane molecules CD40 and HLA-DR on DCs were detected by flow cytometry(FCM).Results The eukaryotic expressive plasmid pcDNA4/Myc-His-LL-37 was constructed successfully and it expressed in eukaryotic cells HEK293.FCM results indicated that the expressions of CD40 and HLA-DR on the membrane of DCs which were stimulated by the supernatant produced by pcDNA4/Myc-His-LL-37 were higher than those in control group(P

Objective To evaluate the effects of problem-based learning (PBL) teaching model in medical imaging education in China. Methods Such databases as PubMed, Medline, CNKI, WanFang, VIP Data were electronically searched for literature on PBL versus lecture-based learning (LBL) applied in medical imaging education in China up to April, 2015. According to the strict quality evaluation of the in-cluded studies, meta-analysis was performed using RevMan 5.2 software. Results Fourteen studies were included totally. Studies included 1 233 students, of whom the PBL group had 608 cases, while LBL group had 625 cases. Compared with LBL, PBL was superior in medical imaging theoretical scores [WMD=5.22, 95%CI(3.06, 7.37), P=0.000], and the case analysis scores [WMD=6.45, 95%CI(4.77, 8.12), P=0.000]. PBL was also superior in the autonomous learning ability [RR=1.78, 95%CI (1.47, 2.16), P=0.000], the unity cooperation ability [RR=1.42,95%CI (1.25, 1.61), P=0.000] and analysis ability [RR=1.73,95%CI (1.42, 2.11), P=0.000]. There were significant differences between PBL group and LBL group. Conclusion PBL can improve teaching results in medical imaging education.

Chymosin is an important industrial enzyme widely used in cheese manufacture. To improve expression efficiency of recombinant bovine chymosin in Kluyveromyces lactis strain GG799, we designed and synthesized a DNA sequence encoding bovine prochymosin gene (GenBank Accession No. AA30448) by using optimized codons. The synthesized prochymosin gene was amplified by two-step PCR method, and then cloned into the expression vector pKLAC1, resulting in pKLAC1-Prochy. pKLAC1-Prochy was linearized and transformed into K. lactis GG799 by electrotransformation. Positive clones were screened by YEPD plates containing 1% casein. A recombinant strain chyl with highest activities and multi-copy integration which was detected by using specifical integration primers was chosen and fermented in flask. Prochymosin was expressed in K. lactis successfully. SDS-PAGE analysis revealed that the purified recombinant bovine prochymosin had a molecular mass of 41 kDa. After acid treatment, molecular weight of chymosin is about 36 kDa, the same as native bovine chymosin. Activity tests showed that the chymosin activity of the culture supernatant was 99.67 SU/mL after 96 h cultivation. The activities of chymosin were not prominent increased when galactose was used as carbon source instead of glucose, which proved that the fermentation of recombinant strain does not need galactose inducing. The recombinant K. lactis strain obtained in this study could be further used to produce recombinant chymosin for cheese making.
Animals ; Cattle ; Chymosin ; biosynthesis ; genetics ; Enzyme Precursors ; biosynthesis ; genetics ; Gene Expression Regulation, Fungal ; genetics ; Genetic Vectors ; genetics ; Kluyveromyces ; genetics ; growth & development ; metabolism ; Protein Engineering ; Recombinant Proteins ; biosynthesis ; genetics

Objective:To investigate the correlation of the expression of forkhead transcription factor O1 (FOXO1) with clinicopathological features and the prognosis in patients with diffuse large B-cell lymphoma (DLBCL).Methods:The data of 42 patients newly diagnosed with DLBCL in Hebei General Hospital admitted from June 2012 to January 2020 were collected. The expressions of FOXO1, phosphorylated FOXO1 (p-FOXO1) in DLBCL tissues were detected by using immunohistochemistry. The association of FOXO1 expression with clinicopathological features and the prognosis in DLBCL patients was retrospectively analyzed.Results:The positive rate of FOXO1 was 42.9% (18/42) and the positive rate of p-FOXO1 was 28.6% (12/42) in DLBCL tissues. There were no statistically significant differences in the positive rates of FOXO1 and p-FOXO1 among patients stratified by gender, age, Ann Arbor staging, immunophenotype, Eastern Cooperative Oncology Group score, lactate dehydrogenase, international prognostic index, β 2-microglobulin (β 2-MG) and primary sites (all P > 0.05). The positive rate of FOXO1 in patients with non-B symptoms was higher than that in those with B symptoms [53.6% (15/28) vs. 21.4% (3/14), χ2=3.938, P=0.047], and there was no statistically significant difference in the positive rate of p-FOXO1 among patients with or without B symptoms ( P > 0.05). The 2-year overall survival (OS) rate in FOXO1 positive group was higher than that in FOXO1 negative group (90.9% vs. 66.7%), the 2-year OS rate in p-FOXO1 positive group was lower than that in p-FOXO1 negative group (50.0% vs. 85.0%), and the differences were not statistically significant (all P > 0.05). Among patients without B symptoms, the 2-year OS rate in FOXO1 positive group was higher than that in FOXO1 negative group (100.0% vs. 50.0%, χ2=5.486, P=0.019). Among patients with primary lymph node, elevated β 2-MG and non-B symptoms, the 2-year OS rate in p-FOXO1 negative expression group was higher than that in p-FOXO1 positive group (100.0% vs. 50.0%, 100.0% vs. 25.0%, 91.7% vs. 33.3%), and the differences were statistically significant (all P < 0.05). Conclusions:FOXO1 may be involved in the development and progression of DLBCL, and FOXO1 positive expression may indicate the good prognosis of patients. These results suggest that p-FOXO1 positive expression may be related with poor prognosis.

To provide new ideas for clinical diagnosis and treatment of coronavirus disease 2019 (COVID-19), this study explore the expression level and prognostic value of platelet parameters in mild, moderate and severe COVID-19. This is a retrospective analysis. From January to May 2020, a total of 69 patients who were diagnosed with COVID-19 in the Third Central Hospital and the Jinnan Hospital (both situated in Tianjin) were enrolled in the disease group. According to the severity, these patients were divided into mild group (15 cases), moderate group (46 cases), and severe group (8 cases). In the same period, 70 non-infected patients were enrolled in control group. The level of white blood cell count (WBC), absolute neutrophil count (NEU#), absolute lymphocyte count (LY#), neutrophil-lymphocyte ratio (NLR), red blood cell count (RBC), hemoglobin (Hb), platelet count (PLT), mean platelet volume (MPV), platelet distribution width (PDW), and platelet-large contrast ratio (P-LCR) before and after treatment were analyzed. Binary logistic regression analysis is used to establish a mathematical model of the relationship between these indexes and the outcome of severe COVID-19 patients. The receiver operating characteristic(ROC) curve is used to further explore the prognosis value of MPV, P-LCR, NLR separately and jointly in COVID-19 patients. Compare to the control group, WBC and NE# increase ( Z=-5.63, P<0.01; Z=-9.19, P<0.01) and LY# decrease ( Z=-9.34, P<0.01) in the severe group; NLR increase with the aggravation of the disease, there is significant difference between groups ( Z=17.61, P<0.01); PLT, PDW, MPV and P-LCR decrease with the aggravation of the disease, there is significant difference between groups ( Z=9.47, P<0.01; Z=11.41, P<0.01; Z =16.76, P<0.01; Z=13.97, P<0.01). Binary logistic regression analysis shows MPV, P-LCR and NLR have predictive value for severe COVID-19 patients. There is a negative correlation between MPV, P-LCR and severe COVID-19 patients ( OR=1.004, P=0.034; OR=1.097, P=0.046). There is a positive correlation between NLR and severe COVID-19 patients ( OR=1.052, P=0.016). MPV and P-LCR of patients with good prognosis after treatment were significantly higher than those before treatment ( Z=-6.47, P<0.01; Z=-5.36, P<0.01). NLR was significantly lower than that before treatment ( Z=-8.13, P<0.01). MPV and P-LCR in poor prognosis group were significantly lower than those before treatment ( Z=-9.46, P<0.01; Z=-6.81, P<0.01). NLR was significantly higher than that before treatment ( Z=-3.24, P<0.01). There were significant differences between good and poor prognosis groups before and after treatment in MPV, P-LCR and NLR ( P<0.01). Combination of these three indexes, ROC shows the AUC is 0.931, the sensitivity is 91.5%, the specificity is 94.1%, the positive predictive value is 88.9%, and the negative predictive value is 87.4%, which is better than any of these indexes separately. Changes in these parameters are closely related to clinical stage of COVID-19 patients. MPV, P-LCR and NLR are of great value in the prediction and prognosis of severe COVID-19 patients.