Production internship is an important teaching tache for undergraduate students to carry out engineering training by using professional skills, and it is a key starting point for fostering application-oriented talents in biotechnology. The Course Group of 'production internship of biotechnology majors' of Binzhou University is investigating application-oriented transformation for local regular colleges and universities, as well as fostering high-level application-oriented talents. By taking green fluorescent protein (GFP) polyclonal antibody as an example, the reform and practice on teaching content, teaching mode, assessment method, continuous improvement of curriculum were carried out. Moreover, the characteristics of the Yellow River Delta-Binzhou Biotechnology & Pharmaceutical Industrial Cluster were taken into account to intensify academic-enterprise cooperation. On one hand, this Course Group designed and rearranged the course contents, carried out essential training through online resources and platforms such as virtual simulation, and recorded, tracked and monitored the progress of production internship through practical testing and software platforms like 'Alumni State'. On the other hand, this Course Group established a practice-and application-oriented assessment method in the process of production internship and a dual evaluation model for continuous improvement. These reform and practices have promoted the training of application-oriented talents in biotechnology, and may serve as a reference for similar courses.
Humans ; Internship and Residency ; Curriculum ; Students ; Biotechnology

Objective:To explore the effect of alcohol abstinence intervention based on timing theory on patients with alcoholic liver disease.Methods:A total of 106 patients with alcoholic liver disease hospitalized in the Department of liver disease of Taian Medical District, 960th Hospital of Chinese PLA from July 2018 to June 2019 were selected by convenience sampling method and divided into observation group and control group by random digits table method, 53 cases in each group. The control group received routine nursing, and through the improvement of patients' cognition and support system, implemented short abstinence intervention during hospitalization; the observation group received abstinence intervention based on timing theory on the basis of the control group intervention. At 1 month and 6 months after discharge, the differences of rehydration rate, alcohol dependence and physical and mental status between the two groups were compared.Results:Finally, 49 cases in the control group completed the study, and 51 cases in the observation group completed the study. The rehydration rates of the observation group were 21.57%(11/51) and 15.69%(8/51) respectively at 1 month and 6 months after discharge, while those of the control group were 40.82%(20/49) and 36.73%(18/49) respectively at 1 month and 6 months after discharge. The difference was statistically significant ( χ2 values were 4.328, 5.754, P<0.05). The alcohol dependence scores were 0(2,3), 0(1,2) in the observation group and 2(0,3), 3(1,4) in the control group at 1 month and 6 months after discharge, and the difference was statistically significant ( Z values were -6.719, -7.345, P<0.01). There was no significant difference in the score of Symptom Checklist-90(SCL-90) before intervention and 1 month after discharge between the two groups ( P>0.05). Six months after discharge, the score of SCL-90 was 8.26 ± 1.37 in the observation group and 10.11 ± 1.68 in the control group, and the difference was statistically significant( t value was 6.046, P<0.01). Conclusions:The application of timing theory in alcohol abstinence of patients with alcoholic liver disease can significantly reduce the relapse rate and the degree of alcohol dependence of patients with alcoholic liver disease, improve the physical and mental state of patients.

Clinically, a large proportion of glaucoma patients undergo repeated intraocular pressure (IOP) spike (Spike IOP) attacks during their sleep, which may facilitate retinopathy. In this study, we established a mouse model of repeated transient Spike IOP to investigate the direct damage to the retina following Spike IOP attacks, and elucidated the underlying molecular mechanism. We analyzed the changes in the number of retinal ganglion cells (RGCs) via immunofluorescence. Thereafter, we detected retinal cell apoptosis via terminal deoxynucleotidyl transferase deoxyuridine triphosphate (dUTP) nick-end labeling (TUNEL) staining, and performed RNA sequencing (RNA-seq) to reveal the underlying molecular mechanism. Finally, we validated the expression of key molecules in the endoplasmic reticulum (ER) stress pathway using quantitative real-time polymerase chain reaction (qRT-PCR) and western blot analysis. Results revealed a time-dependent RGC loss in Spike IOP, evidenced by a reduction in the number of Brn3a-positive RGCs in experimental eyes following a 7-d continuous treatment with Spike IOP. In addition, TUNEL staining indicated that apoptosis of retinal cells started in the outer nuclear layer (ONL), and then spread to the ganglion cell layer (GCL) with time. RNA-seq analysis revealed that ER stress might be involved in Spike IOP-induced retinal injury. This result was corroborated by western blot, which revealed upregulation of ER stress-related proteins including binding immunoglobulin protein/glucose-regulated protein 78 (BiP/GRP78), phosphorylated inositol-requiring enzyme 1 (p-IRE1), unspliced X-box-binding protein 1 (XBP1-u), spliced X-box-binding protein 1 (XBP1-s), phosphorylated c-Jun N-terminal kinase (p-JNK), C/EBP-homologous protein (CHOP), and B-cell lymphoma 2 (Bcl-2)-associated X protein (Bax). These findings indicate that repeated IOP transients are detrimental to the retina, while ER stress plays an important role in retinal cell apoptosis in this situation. Notably, repeated Spike IOP among glaucoma patients is a crucial factor for progressive retinopathy.

OBJECTIVEThis study is performed to investigate the cell topographies and biomechanical properties of two different types of temporomandibular joint (TMJ) discs from goats by using JPK Nano Wizard 3 biological atomic force microscopy (AFM). This process provides a guideline for selecting seed cells for TMJ disc tissue engineering.

METHODSTMJ disc cells from primary goats were cultured by monolayer culture method. AFM was used to contact scan the topographies of the two types of TMJ disc cells under physiological environment. Approximately 20 chondrocyte-like and fibroblast-like cells were selected randomly to plot the force-versus-distance curves of the cytoplasm and nucleus. Young's modulus and adhesion were analyzed by JPK Data Processing.

RESULTSThe triangle-shapednucleus of the chondrocyte-like cell occupied a large portion of the cell. Cytoskeleton was arranged dendritically on the surface. Pseudopodia were extended from cell edges. The spindle-shaped nucleus of the fibroblast-like cell occupied a significantly larger region compared with the cytoplasmic region. Cytoskeleton was arranged regularly. Cell edges were smooth with less pseudopodia extended. No difference was found in the surface roughness between the two types of cells. According to the force-versus-distance curves, the Young's moduli of the two types of cells were not statistically different (P>0.05), but differences were found in the cytoplasmic regions (P=0.047). No statistical difference was found in the adhesions between the two types of cells (P>0.05).

CONCLUSIONThe AFM topography and curves were compared and analyzed. The two types of TMJ disc cells exhibited significantly different topographies, but only slight difference in their mechanical abilities.

Objective??This?study?is?performed?to?investigate?the?cell?topographies?and?biomechanical?properties?of?two?different?types?of?temporomandibular?joint?(TMJ)?discs?from?goats?by?using?JPK?Nano?Wizard?3?biological?atomic?force?microscopy?(AFM).?This?process?provides?a?guideline?for?selecting?seed?cells?for?TMJ?disc?tissue?engineering. Methods??TMJ?disc?cells?from?primary?goats?were?cultured?by?monolayer?culture?method.?AFM?was?used?to?contact?scan?the?topographies?of?the?two?types?of?TMJ?disc?cells?under?physiological?environment.?Approximately?20?chondrocyte-like?and?fibroblast-like?cells?were?selected?randomly?to?plot?the?force-versus-distance?curves?of?the?cytoplasm?and?nucleus.?Young’s?modulus?and?adhesion?were?analyzed?by?JPK?Data?Processing.?Results??The?triangle-shapednucleus?of?the?chondrocyte-like?cell?occupied?a?large?portion?of?the?cell.?Cytoskeleton?was?arranged?dendritically?on?the?surface.?Pseudopodia?were?extended?from?cell?edges.?The?spindle-shapednucleus?of?the?fibroblast-like?cell?occupied?a?significantly?larger?region?compared?with?the?cytoplasmic?region.?Cytoskeleton?was?arranged?regularly.?Cell?edges?were?smooth?with?less?pseudopodia?extended.?No?difference?was?found?in?the?surface?roughness?between?the?two?types?of?cells.?According?to?the?force-versus-distance?curves,?the?Young’s?moduli?of?the?two?types?of?cells?were?not?statistically?different?(P>0.05),?but?differences?were?found?in?the?cytoplasmic?regions?(P=0.047).?No?statistical?difference?was?found?in?the?adhesions?between?the?two?types?of?cells?(P>0.05).?Conclusion??The?AFM?topography?and?curves?were?compared?and?analyzed.?The?two?types?of?TMJ?disc?cells?exhibited?significantly?different?topographies,?but?only?slight?difference?in?their?mechanical?abilities.

BACKGROUND:Our preliminary studies have shown that basic fibroblast growth factor can induce the differentiation of bone marrow mesenchymal stem cells into disc cells of the temporomandibular joint, and for basic fibroblast growth factor, 10μg/L is superior to 5μg/L in col agen synthesis.
OBJECTIVE:To observe ultrastructural changes of bone marrow mesenchymal stem cells after being induced by different concentrations of basic fibroblast growth factor.
METHODS:We cultured primary sheep bone marrow mesenchymal stem cells and selected passage 3 and 4 cells at good growth state. Bone marrow mesenchymal stem cells were stimulated with 5 and 10μg/L basic fibroblast growth factor and their growth state was observed under inverted phase contrast microscope. Uninduced cells served as controls. The slides with cellcrawling pieces were stained with Safranin O, picrosirius and type I col agen immunohistochemistry at days 7, 14 and 21, respectively. Simultaneously we col ected the cells at day 21 to observe the ultrastructural changes of bone marrow mesenchymal stem cells.
RESULTS AND CONCLUSION:After being induced with different concentrations of basic fibroblast growth factor, bone marrow mesenchymal stem cells were able to differentiate into disc cells of the temporomandibular joint;and after being induced with 10μg/L basic fibroblast growth factor, cells were more like fibroblast-like cells of the temporomandibular joint disc. These findings indicate that bone marrow mesenchymal stem cells have morphological basis for differentiation to the fibroblast-like cells of the temporomandibular joint disc.

BACKGROUND:Early detection and accurate staging diagnosis of heart failure are the basis of good clinical therapy efficacy. Due to lack of simple and effective staging model for the diagnosis of heart failure, it is difficult to diagnose heart failure in clinics, leading to poor control of heart failure. OBJECTIVE:To establish the disease staging model based on Adaboost and SVM for heart failure, and improve the accuracy of diagnosis and staging of heart failure. METHODS:A total of 194 cases were roled into this study, including heart failure patients and healthy physical examination persons. According to the stage standards formulated by American Colege of Cardiology and American Heart Association, specific clinical feature parameters closely related to heart failure were colected and selected. Based on clinical diagnosis results and using Adaboost model and SVM model, we trained the models for heart failure diagnosis and staging, thus obtaining diagnosis model. RESULTS AND CONCLUSION: The parameters included stroke volume, cardiac output, left ventricular ejection fraction, left atrial diameter, left ventricular internal diameter at end-systole, N-terminal pro-brain natriuretic peptide and heart rate variability. As for the Adaboost model, its sensitivity and specificity was 100% and 94.4%, respectively. At the same time the SVM model had good sensitivity and specificity, 86.5% and 89.4% respectively. Adaboost classification model can be accurate in the diagnosis of heart failure symptoms, the accuracy reached 89.36%. On the basis of the diagnosis of heart failure, the SVM classification model is effective in staging the severity of heart failure, staging accuracy for staging B and C was 86.49% and 81.48%, respectively. The findings indicate that, combining Adaboost and SVM machine learning models could provide an accurate diagnosis and staging model for heart failure.

Objective To observe the correlation between vitamin D and liver cirrhosis classification in hepatic diabetes.Methods Thirty-eight patients with hepatic diabetes,48 patients with type 2 diabetes and 30 cases of healthy controls were collected.The level of serum 25-(OH)D3 was determined,and the Child-Pugh score was calculated.The correlation between pathogenesis and pathogenetic condition of hepatic diabetes was analyzed.Results The levels of serum 25-(OH)D3 in hepatic diabetes patients,type 2 diabetes patients and healthy controls were (38.23 ± 12.47),(63.33 ± 13.58) and (86.14 ± 16.25) μ mol/L,and there was statistical difference (P ＜ 0.05).There was negative correlation in hepatic diabetes between serum 25-(OH)D3 level and liver cirrhosis Child-Pugh score (r =-0.363,P ＜ 0.05).Conclusion Low level of vitamin D may be one of the pathogenesis of hepatic diabetes.

Objective To observe the role of mulberry flavone on the expression of insulin receptor substrate(IRS)in HepG2 cells with insulin resistance, and to explore the possible molecular mechanism that mulberry flavone improves insulin resistance. Methods HepG2 cells were cultured to establish insulin-resistance cell model in high concentration of insulin, and then incubated with mulberry flavone. Effect of mulberry flavone on the HepG2 cell model of glucose incorporation rate was observed. Western blot was used to observe the variety of IRS protein expression. Results Mulberry flavone increased the glucose incorporation rate of (33. 9 ± 1.0)higher than that of Conclusions Mulberry flavone can improve insulin resistance. Furthermore, IRS protein expression increasing is the possible molecular mechanism that mulberry flavone improves insulin resistance.