Objective To investigate the relationship of gene transfection efficiency with different ultrasound exposure time and different dose of microhuhble,and to find the appropriate ultrasound parameters for gene transfection. Methods Plasmid encoding enhanced green fluorescent protein(pEGFP) was chosen as a report gene and HepG2 cells were chosen as research object. The HepG2 cells plus pEGFP and different dose of microbubble were exposed to ultrasound(1 MHz,0.5 W/cm2) with varying time. Twenty-four hours later, the expression of EGFP in the cells was observed by fluorescence microscope,the transfection efficiency was assessed by FACS and the cell viability was observed by trypan blue exclusion. Results The expression of EGFP in all experimental groups was different,and the approving transfection efficiency was got by ultrasound exposed for 20 s when the dose of microbubble was 60 μl. Conclusions With fixed ultrasound frequency and power, different transfection efficiency was got when the exposure time and dose of microbubble were different. The appropriate parameter was 20 s,60 μl, which can supply information for further study.

OBJECTIVE To evaluate the protective effect of Agaricus bisporus intracellular polysaccharides(IPS) and exopolysaccharides (EPS) on immunological liver injury induced by concanavalin A (Con A). METHODS Mice were pretreated with IPS and EPS (100, 200 and 400 mg kg- 1, ig) daily for 12 d. Immunological liver injury was induced by Con A 25 mg·kg-1 byinjection via the tail vein of mice.Eight hours after injection of Con A, the indexes of the liver, spleen and thymus, serum level of glutamicpyruvic transaminase (GPT), glutamic-oxalacetic transaminase (GOT), tumor necrosis factor-α (TNF-α) and interferon- γ (IFN- γ), splenic lymphocyte percentages of CD4 + and CD8 + , and liver homogenate content of superoxide dismutase (SOD) and malondialdehyde (MDA) were measured. Liver pathological changes were observed by HE staining. RESULTS Compared with normal group, the autoimmune liver injury in mice induced by Con A resulted in an increase in the liver index (P<0.01) , spleen index (P<0.01), the activity of GPT (P<0.01) and GOT (P<0.01), the content of TNF- α (P<0.01) and IFN- γ (P< 0.01), and the level of MDA (P<0.01), but a decrease in the thymus index (P<0.01), the percentage of CD4+ (P<0.01) , the ratio of CD4+/CD8+ (P<0.01), and SOD activity (P<0.01). Compared with model group, treatment with IPS (200 and 400 mg·kg-1) and EPS (200 and 400 mg·kg-1 ) respectively resulted in an increase in the thymus index (P<0.01) but in a decrease in the liver index and spleen index (P<0.01). Similarly, the activity of GOT and GPT was decreased obviously (P<0.01), and the content of TNF-α and IFN-γ in IPS and EPS 200 and 400 mg·kg-1 groups was decreased. Compared with model group, the activity of SOD in IPS and EPS (200 and 400 mg·kg- 1) group was increased (P<0.01) while MDA was decreased (P<0.01). Moreover, the percentage of CD4 + Iymphocytes decreased (P<0.01), whereas no significant difference was found in the ratio of CD4 +/CD8 + .Pathological changes of the liver were observed under a microscope. Pretreatment with IPS and EPS could effectively reduce the liver injury induced by Con A. CONCLUSION IPS and EPS have certain protective effect on immunological liver injury, which may be related to their ability to clean up free radicals, control lipid peroxidation and regulate the balance of the immune system.

ObjectiveTo explore the attentional biases in male alcohol dependent (AD) patients and the correlations between the attentional bias and alcohol-related factors.MethodsA total of 30 recently detoxified male individuals with alcoholism were compared with 37 healthy controls ( HC ) on the Chinese Emotional Stroop Task using negative,neutral,and alcohol-related words.ResultsThe comparison between AD group( ( 1382.13 ±323.38) ms,( 1365.76 ±313.03)ms,( 1433.20 ±342.23) ms,respectively) and HC group( (797.27 ±216.97)ms,( 794.11 ± 209.41 ) ms,(799.40 ± 215.82 ) ms respectively) on the reaction time of neutral,negative and alcohol-related words were significant ( t =8.822,P ＜ 0.001 ; t =8.922,P ＜ 0.001 ; t =9.234,P ＜ 0.001 ).The error number of of the neutral and negative- related words of the patients ( ( 3.70 ± 2.56) time,( 4.23 ± 2.53 ) time)was worse than that of HC group( ( 2.11 ± 1.87 ) time,( 1.92 ± 1.82 ) time) ( t =2.939,P =0.005 ; t =4.355,P ＜0.01 ).Error number of the alcohol- related words between two groups were not significant;Its alcohol-related words attentional bias negative correlation to the age of initial alcohol use(P＜ 0.05 ),and positive correlation to continue drinking and score of self-rating depression scale (P＜0.05).Age of addiction and the score of self-rating anxiety scale enter the regression equation of alcohol-related words.ConclusionThese results demonstrate that alcoholics have attentional bias in alcohol-related words and reward-related brain regions may be associated with craving among male patients with attentional bias.

Objective To explore the application value of circulating tumor cell (CTC) counting in the diagnosis of patients with colorectal cancer.Methods From July to October in 2015,a total of 61 patients firstly diagnosed with colorectal cancer,at stagc] to Ⅳ were enrolled.At the same period,20 individuals (healthy volunteers or patients with benign colorectal diseases) were selected as controls.Peripheral blood (7.5 mL) was taken before therapy.Peripheral blood CTC were counted by immunomagnetic beads enrichment combined with fluorescent staining method in two hours.The positive detection rate of CTC for colorectal cancer,especially early colorectal cancer was analyzed.And the priority of combination it with carcino-embryonic antigen (CEA) was also investigated.Chi-square test,t test and rank-sum test were used for statistical analysis.Results Two patients could not be pathological staged,due to the lack of whole body imaging evaluation.There were 30 patients with colorectal cancer at early stage (stage Ⅰ to Ⅱ) and 29 patients at advanced stage (stage Ⅲ to Ⅳ).The difference in CEA level between patients at early stage and advanced stage ((3.3±1.1) μg/L vs (20.4±3.3) μg/L) was statistically significant (t=-2.74,P=0.008).The median cell number of CTC of colorectal cancer group and control group was 1 (interquartile range=4) and 0,respectively,and the difference was statistically significant (Z=2.721,P=0.007).Taken 0/7.5 mL and 5 μg/L as the cut-off value of CTC and CEA,the detection rates of CTC for colorectal cancer and early colorectal cancer were 65.6％ (40/61) and 63.3％ (19/30),respectively;the detection rates of CEA for colorectal cancer and early colorectal cancer were 29.5 ％ (18/61) and 13.3％ (4/30).When CTC combined with CEA,the detection rates for colorectal cancer and early colorectal cancer were 73.8％ (45/61) and 66.7％ (20/30),respectively.Conclusions CTC has certain detection value in colorectal cancer especially early colorectal cancer.If it combined with CEA,the detection rate is much higher.

Comprehensive experiments course is a bridge for higher vocational students to integrate theoretical knowledge with production practice. The article introduces that our biological pharmacy department is committed to the principles of "promotion of teaching, learning and construction through skills competition so as to integrate education and training". By taking penicillin fermentation process as an example, reform has been made in several aspects including teaching objectives, teaching content and teaching methods. We integrate the practical operation of fermentation equipment with virtual simulation software to develop a two-way interactive course. By reducing the subjective dependence, the quantitative management and evaluation of fermentation process parameter control were put into place, which efficiently integrated the skills competition with practical teaching. Improved teaching performance has been achieved over recent years, which may facilitate the reform and practice of similar courses based on skills competition.
Humans ; Clinical Competence ; Learning ; Students ; Technology ; Biological Products