Objective To construct one high-copy BAC vector with cloning capacity for large DNA fragment insertion and use it for the haplotype analysis of CYP2A6 gene.Methods Oligos including multiple cloning sites were annealed and ligated into the Hind Ⅲ/BamH Ⅰ site of pGEM-3Z and pBeloBACll,separately.Then,the intermediate vectors were digested by Hind Ⅲ and ligated together,so as to get the head-to-tail oriented high-copy BAC vector pBAC-BJH after the blue-white spotting test.STI PCR method was used for the amplification of whole CYP2A6 gene,and purified amplicon was double digested by BstB Ⅰ/Mlu Ⅰ and ligated into the same site of newly constructed vector pBAC-BJH.Several clones were then picked up and sequenced for the haplotype analysis of carriers with newly discovered CYP2A6 variant 355A＞T.Results One high-copy BAC vector pBAC-BJH with 7 newly added multiple cloning sites was successfully constructed.High copy number and multiple cloning sites were the advantages of this plasmid.With this vector,haplotype analysis result for 22C＞T,51A＞G,355A＞T in carrier with newly detected CYP2A6mutation was CGT.Conclusion One vector pBAC-BJH convenient for cloning large DNA fragment was successfully developed,and it can be used for the haplotype analysis of cytochrome p450 gene and cloning or sequencing of other genes with large genomic DNA inserts.

Diabetic foot(DF)is one of the most severe chronic complications of diabetes mellitus(DM),and it is an important cause of disability and mortality inpatients with DM.Changes in proteins in the tissues,organs and circulation of organisms can serve as microscopic reflections of the disease development process.Proteomics is an important technologyto explore the pathogenesis and treatment mechanism of dis-ease,and to find potential therapeutic targets and prognostic biomarkers of diseases.This article reviews the progress of proteomics research in DF.

Rheumatoid arthritis is a chronic, systemic autoimmune disease predominantly based on joint lesions with an extremely high disability and deformity rate. Several drugs have been used for the treatment of rheumatoid arthritis, but their use is limited by suboptimal bioavailability, serious adverse effects, and nonnegligible first-pass effects. In contrast, transdermal drug delivery systems (TDDSs) can avoid these drawbacks and improve patient compliance, making them a promising option for the treatment of rheumatoid arthritis (RA). Of course, TDDSs also face unique challenges, as the physiological barrier of the skin makes drug delivery somewhat limited. To overcome this barrier and maximize drug delivery efficiency, TDDSs have evolved in terms of the principle of transdermal facilitation and transdermal facilitation technology, and different generations of TDDSs have been derived, which have significantly improved transdermal efficiency and even achieved individualized controlled drug delivery. In this review, we summarize the different generations of transdermal drug delivery systems, the corresponding transdermal strategies, and their applications in the treatment of RA.

The rapid development of bioeconomy urgently needs the support of biotechnology talents. Establishing an innovative training mode of biotechnology talents can provide support for regional economic development and industrial upgrading. Closely revolved around the concepts of new engineering disciplines development, such as serving the national strategy, docking industry, leading the future development and student-centered, a new economy-oriented training system was developed in School of Bioengineering of Dalian University of Technology. These systems include interdisciplinary curriculum system reconstruction, project-based teaching mode reform, evaluation system implementation and other aspects. The reform and exploration of the first-class biotechnology major under the new economic situation, puts forward the theory of value guidance, deep foundation, strong sense of innovation, technical and non-technical core ability literacy. This reform meets the industry demand for talent diversification, personalization, and dynamic change, helps the merge of industry and education, which provides a way for fostering first-class biotechnology-majored undergraduates.
Humans ; Biotechnology ; Bioengineering ; Biomedical Engineering ; Students ; Curriculum

Objective: To construct a scientific behavioral research of pasta matrix reaching task (PMRT) and comprehensively evaluate sensory-motor dysfunction caused by brain injury. Methods: Twenty-one SD rats were subjected to 14-days pasta matrix grasping training and then were randomly divided into model group (11 rats) and sham group (6 rats). Motor cortex ischemia was induced by injection of endothelin-1 in SD rats. The number of pasta grabed by the injured forelimb and the location in matrix were evaluated daily 7 days after surgery. The infarct volume was measured by Nissl staining at the 7 days, 14 days, and 28 days after stroke. Results: The number of pasta obtained by rats was reduced from (33.43±1.02) to (20.57±0.57) at 7 days post stroke in model group, and then increased to (26.85±0.98) at 28 days post stroke, although there was a significant difference between sham group(32.33±1.45) and ischemic group (t=3.198, P<0.05). The frequency of retrieval from each slot of the pasta matrix represented that sham group demonstrated a significant gain in performance in the antero quadrant of the matrix compared to ischemic rats by the fourth week after stroke.The stroke volume was decreased from (37.82±1.17)mm³ at 7 days post-stroke to (24.35±0.38)mm³ at 28 days post-stroke, indicating brain recovery from ischemic injury. Conclusion The pasta matrix reaching task can function as a versatile and sensitive behavioral assay that permits experimenters to collect accurate outcome data and manipulate limb use to mimic human clinical phenomena including compensatory strategies and focused rehabilitative training after stroke.

Objective To evaluate the biodistribution and radiation-absorbed doses of main organs in healthy people with 13 N-ammonia.Methods Five healthy volunteers underwent whole-body PET and CT scans after injection of 666-814 MBq of 13 N-ammonia.The serial dynamic emission images of each healthy volunteer were acquired.ROI were drawn manually based on the transverse CT images and transferred to the corresponding PET slices.Radiation-absorbed doses were calculated using the medical internal radiation dosimetry (MIRD) method.Results The highest concentrations of 13 N-ammonia were found in the heart,liver and kidneys,followed by pancreas,brain,spleen and stomach.The organ of highest absorbed dose was heart with (7.14±3.63) × 10-3 mGy/MBq.The whole-body absorbed dose was (2.11±0.44) × 10 3 mGy/MBq.The whole-body effective dose was (6.58± 1.23) × 10-3 mSv/MBq.Conclusion As one of the most important myocardial perfusion tracers,the whole-body 13 N-NH3 · H2 O PET appears to be safe for humans.

Humans ; Lymphoma, Mantle-Cell ; diagnosis ; Otolaryngology

OBJECTIVE: To investigate the application of endoscopic nasal lateral wall dissection in lesions of the maxillary sinus. METHOD: Ten hospitalized patients with the maxillary sinus lesions were treated with the endoscopic nasal lateral wall dissection. RESULT: All 10 patients were unilateral invasion. Among them, 7 cases were inverted papilloma, 2 cases were recurrent antrochoanal polyps, 1 case was sinusal tooth. The tumors and antrochoanal polyps originated from the every part of the maxillary sinus wall during operation, especially from the anterior and media wall. During 10-62 months follow-up,epithelization of nasal occured and the shape of inferior turbinate was well. All of them had no epiphora. CONCLUSION Endoscopic nasal lateral wall dissection can remain the function of nasal lacrimal duct and nasal cavity,and may provide a new minimally invasive approach for complete resection of lesions of nasal cavity and the maxillary sinus.
Dissection ; Endoscopy ; Humans ; Lacrimal Apparatus ; Maxillary Sinus ; pathology ; Nasal Cavity ; Nasal Polyps ; surgery ; Papilloma, Inverted ; surgery ; Turbinates

Objective To explore the expressions and clinical significances of deleted in liver cancer-1 (DLC-1 ) and Rho associated coiledcoil forming protein kinase (ROCK )Ⅰ in non-small lung cancer (NSCLC).Methods The expressions of DLC-1 and ROCKⅠ in NSCLC and adjacent tissue of 48 patients with pathologically confirmed as NSCLC and undergone surgical resection were detected by immunohistochemis-try EnVision method.The correlations among DLC-1 protein,ROCKⅠ protein and the clinical pathological characteristics were analyzed.The prognostic value of DLC-1 in patients with NSCLC was studied.Results The expression of DLC-1 protein in NSCLC tissue was low or missing,and the positive rate was 33.3%(16/48),significantly lower than that in the tissue adjacent to carcinoma 70.8% (34/48),with statistical significance (χ2 =13.523,P<0.01).The positive expression rate of ROCKⅠprotein in NSCLC was 58.3%(28/48),higher than that of tissue adjacent to carcinoma 0(0/48),with statistical significance (χ2 =39.529, P<0.01).The expression of DLC-1 protein was correlated with tumor differentiation,lymph node metastasis and clinical stage,rather than with sex,smoking history and organization type.Through the correlation analy-sis,the expression of ROCKⅠin DLC-1 positive group was 37.5%(6/16),and the expression rate of ROCKⅠ in DLC-1 negative group was 68.8%(22/32).There was negative correlation between DLC-1 and ROCKⅠin NSCLC tissues (r=-2.214,P=0.039).The 3 year survival rate in DLC-1 protein high expression group was obviously higher than that in low expression group,with statistical significance (P=0.043).Conclusion Low or missing expression of DLC-1 and high expression of ROCKⅠ protein may play an important role in the occurrence and development of NSCLC.Detecting the expression of DLC-1 and ROCKⅠprotein may be useful for evaluating the biological behavior and prognosis of NSCLC.

Objective To investigate the changes of sex hormone and androgen receptor levels and evaluate the relationship of the sex hormones and androgen receptor with coronary heart disease (CHD) in elderly men. Methods A cross-sectional study was performed in 539 elderly men, including 400 healthy people aged 62-92 years and 139 CHD patients aged 60-88 years. The plasma concentrations of total testosterone (TT), free testosterone (FT), dehydroepiandrosterone sulfate (DHEAS), sex hormone binding globulin (SHBG), estradiol (E2), luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were measured. The androgen receptor (AR) level was tested by flow cytometry. Results The fluorescence intensity of DHEAS, TT, SHBG, FT and AR were significantly lower in CHD group than in healthy group (P<0.01);however, FSH and E2 in CHD group were higher(P(0.01). Age was negatively correlated with TT(r=-0.28,P<0.01) and FT (r=-0.17,P<0.05), and positively correlated with SHBG(r=0.14,P<0.05) and E2 (r=0.33, P<0.01). AR fluorescence intensity was negatively correlated with systolic blood pressure (r=-0.12,P<0.01). Logistic regression analysis indicated that TT (OR=1.065,9% CI: 1.012～1.121,P<0.05), SHBG(OR=0.994,95% CI:0.990～0.998,P<0.01) and AR (OR=0.971,95%CI:0.956～0.986, P<0.01)were significantly associated with CHD in elderly male patients. Conclusions The levels of DHEAS, TT, SHBG, FT and AR are lower in elderly men with CHD than in elderly healthy men;however, the FSH and E2 concentrations are higher. Low levels of TT, SHBG and AR may be the independent risk factors for CHD in elderly men.