Objective To observe the changes of molecular markers of prethrombotic state (PTS) in coronary artery disease (CAD) patients with diabetes mellitus (DM) undergoing coronary agioplasty. Methods Thirty-two patients with CAD (group A) and twenty-four CAD patients with noninsulin-dependent diabetes mellitus (NIDDM) (group B) were chosen as objects undergoing coronary agioplasty, and molecular markers of PTS, lipid peroxide (LPO), endothelial function were measured before and 20 minutes, 24 hours, and 7 days after the opration respectively. Results There were significant changes of molecular markers of PTS, LPO, and endothelial function in both groups after the opration, but the changes were even more pronounced in group B. It was on the 7th day that there were still the changes in group B, whereas the changes were prone to the levels of pre-opration in group A. Conclusion There was a series of significant changes of molecular markers of PTS, LPO, and endothelial function in CAD patients after the angioplasty, and the changes were even more pronounced in those with DM.

Objective The tissue engineering technique and the microsurgery technology is combined to construct the uncellular tissue engineering complex with vascularization and membrane guided dual effect. Through comparing study of using the simple biomembrane guided bone regeneration technique to construct the uncellular tissue engineering complex to repair the large segment bone defect in the animal body,the bone reparative effect of the tissue engineering bone wrapped by pedical fascial flap with vessels and that wrapped by the simple biomembrane was compared, thus to provide experimental evidence for the clinical application. Methods Twenty-four Newzland 5-month-old rabbits were used to build the bilateral periosteumincluded bone defect modelsin the middle piece of the ulna and the length of the defect was 1 cm. Autologous red bone marrow was implanted in the tissue engineering bone which was prepared by osteoinductive absorbing material including BMP. The prepared tissue engineering bone was implanted in the bone defect area. The right side was wrapped by the simple absorbable biomembrane, whereas the left side was wrapped by pedical fascial flap with blood supply. At the fourth, eighth, twelfth and sixteenth week after the operation each group was examined by the radiograph (x-ray), the light density measurement, gross morphology and histological inspection,bone shape measurement analysis in the repairing area and the biomechanics measurement at the twelfth week. The data was analyzed to test the difference of the bond defect repair. Results The radiograph, gross morphology and histological inspection showed the growth of vessels in the implant area, the quantity and the forming speed of the bone trabecula and, the cartilaginous tissue, the formation of the mature bone structure,remodeling of the diaphysis, recanalization of the cavum ossis and the absorption and the degradation of the implant of the group of pedical fascial flap with blood supply was superior to that of the group of the simple absorbable biomembrane. At the fourth, eighth, twelfth and sixteenth week after the operation the bone trabecula area were( 20. 35 ± 2. 41 ) %, ( 40. 21 ± 1.97 ) %, (66. 67 ± 3.44 ) % and ( 86. 47 ± 3.99) % respectively in the group of pedical fascial flap with blood supply, and were ( 7. 46 ± 2.64 ) %, ( 20. 66 ± 2. 28 ) % , ( 40. 22 ±1.84)% and(58. 18 ± 1.79) respectively in the group of the simple absorbable biomembrane. At the same time point after the operation the light density were 0. 636 ± 0. 012,0. 596 ± 0. 062,0. 552 ± 0. 009 and 0. 451 ±0. 008 respectively in the group of pedical fascial flap with blood supply, and 0. 742 ± 0. 032,0. 713 ± 0. 022,0. 655 ±0. 018 and 0. 606 ±0. 015 respectively in the group of the simple absorbable biomembrane. The units of blood vessel reproductive area in the bone repair junctional zone were ( 18.75 ± 2. 09 ) %, ( 37.41 ± 3.22 ) %,(53. 06 ±2. 18)% and (36.72 ±4. 73)% respectively in the group of pedical fascial flap with blood supply,and (5. 34 ± 1.17 ) %, (9. 48 ± 2.96) %, ( 22.43 ± 2. 21 ) % and ( 26. 27 ± 3. 14 ) % respectively in the group of the simple absorbable biomembrane. The biomechanics intension was 26.62 ± 3.96 in the group of pedical fascial flap with blood supply and 18. 38 ±0. 71 in the group of the simple absorbable biomembrane at the twelfth week after the operation. All of the differences were significant( P ＜0. 05 ). Conclusion The pedical fascial flap with blood supply has significant effect in promoting the tissue engineering bone to vascularize and promoting the bone formation by vascularization. The membrane guided bone regeneration technique restricted not only the growth of the fibrous connective tissue in the reparative process of the large segment bone defect effectively, but also the ability of fast and effective vascularization, thus the chronic creep and substitution process would be needed. Simple application of the biomembrane can compensate the shortcoming of chronic creep of the implanted material by the growth of the external callus.

Objective To investigate the therapeutic effects of givinostat , a histone deacetylase inhibitor (HDACI), on the development of chronic asthma with airway inflammation , airway remodeling and airway hyperresponsiveness ( AHR) .Methods BALB/C mice were randomly divided into control group , asthma group, dexamethasone group and givinostat group (n=12 per group).AHR was assessed.Total cell numbers and differential counts , interleukin-4 ( IL-4 ) , interleukin-5 ( IL-5 ) and interferon-γ( IFNγ) levels in the bronchoalveolar lavage fluid ( BALF) were measured in the above 4 groups.The pathology of lung tissue was evaluated .Immunohistochemical ( IHC) staining and Western blot were used to detect αsmooth muscle actin(α-SMA) and transforming growth factor-β1(TGFβ1).Results Compared with the asthma only group, givinostat treatment relieved airway resistance (2.96 ±1.01 vs 6.50 ±0.79,P<0.05).Total inflammatory cells [(33.04 ±5.62) ×104/ml vs (98.04 ±9.27) ×104/ml,P<0.01], eosinophil cells [(9.17 ±2.33) ×104/ml vs(37.64 ±6.98) ×104/ml, P <0.01], IL-4 [(10.12 ±2.98)ng/ml vs (16.88 ±2.78)ng/ml,P<0.05] and IL-5 [(27.09 ±3.62)ng/ml vs (37.86 ±7.34)ng/ml, P<0.05] levels were all reduced in givinostat group , while IFNγ[ ( 91.86 ±23.73 ) pg/ml vs ( 60.49 ±11.88 ) pg/ml, P>0.05] was enhanced in BALF.Inflammatory cell infiltration around the airway was reduced , with decreased inflammatory cell score [(1.60 ±0.69) points vs (3.40 ±0.68) points, P <0.01] and inflammatory cell number (111.65 ±31.41 vs 601.25 ±186.85, P<0.01).The goblet cell metaplasia [(26.36 ±2.33)%vs (57.21 ±11.56)%] and collagen deposition area [(52.77 ±7.58)μm2/μm vs (111.81 ±12.40)μm2/μm] were obviously reduced (P<0.01).The expressions of α-SMA and TGFβ1 in the lung tissue were both significantly decreased ( P<0.01 ) .Conclusion Givinostat treatment can reduce airway inflammation , airway remodeling and airway hyperresponsiveness in chronic asthma .Its effect is comparable to that of glucocorticoid hormone treatment .

Objective To investigate the effects of Tubastatin A Hcl,a selective HDAC6 inhibitor,on the development of chronic asthmatic mice with airway inflammation,airway remodeling and airway hyperresponsiveness. Methods BALB/C mice were randomly divided into control group, asthma group,dexamethasone group and Tubastatin A Hcl group. The airway resistance,total cells and different cells in BALF,IL?4,IL?5,TGF?β1 were detected by ELISA. HE、AB?PAS and Masson trichrome staining were carried out to assess the airway inflammation and remodeling. Immuno?histochemical staining and western blotting were adopted to determine the expression ofα?SMA and TGF?β1. Results After drugs treatment,air?way resistance decreased,and levels of IL?4,IL?5,TGF?β1,total inflammatory cells and eosinophils in BALF were relieved. Meanwhile,inflamma?tory cells infiltration,goblet cells metaplasia and collagen deposition in lung tissue were also reduced,but all of above the effects of dexamethasone were better than Tubastatin A Hcl. The expression ofα?SMA and TGF?β1 in the lung tissue decreased significantly after treatment ,in which Tu?bastatin A Hcl were slightly better than dexamethasone treatment. Conclusion Tubastatin A Hcl can effectively relieve airway inflammation ,air?way remodeling and airway hyperresponsiveness in chronic asthmatic mice ,but its effect of anti?inflammatory is worse than dexamethasone treat?ment,while it is better than dexamethasone in the effect of relief airway remodeling.

Objective To identify the important regulatory elements in the promoter of human CD2 associated protein(CD2AP) by conserved sequence analysis among different species and luciferase functional detection. Methods The promoter sequences of CD2AP from different species were analyzed by BLAST. Plasmids containing different length of deletion mutations of human CD2AP promoter were constructed. Pro-moter activities were tested in 3 kinds of cells from different species by luciferase analysis and were tested in HEK-293 cells treated with all-trans-retinoic acid. Results Homologous sequence comparison in CD2AP promoter area among human, cattle and pig showed that putative specific protein 1 (Sp1) sites and down-stream promoter element (DPE) were highly evolutional]y conserved. Progressive deletion luciferase analysis of DNA fragments revealed similar promoter activity style among 3 different cell lines from 3 different spe-cies, HEK-293, BHK-21 and Vero cells. One basic promoter activity located within 500 bp upstream of ATG. Fragments of further upstream 100 bp or more had drastically 10 times increased promoter activity. Two putative Sp1 sites were in this 100 bp region. All-trans-retinoic acid decreased the luciferase activity of CD2AP promoter. Conclusion Putative Spl sites and DPE have important functions in the promoter activity of CD2AP.

Objective To study the effect of repairing limbs bone defect by using uncellular tissue engineering complex of autolegous red bone marrow wrapped by facial flap with vessels to provide evidence for clinical application. Methods Nineteen cases of limbs bone defect were chosen, among them, 3 cases were with benign bone tumor,6 cases were with open fracture causing bone defect and 10 cases were with bone hypotoxic infection after operation. Took autolegous red bone marrow to prepare uncelluar tissue engineering complex with osteoinduction active material (OAM) containing bone morphogenetic protein (BMP). Prepared a facial flap with capillary network originating from an anonymos vessel adjacent to the bone defect,wrappad the tissue engineering bone and filled the bone defect. On a certain time after the operation, the patients were tested by X-ray. Results All the 19 cases were followed up in 1st,3rd,6th, 10th, 12th and 18th month after the operation. There were 3 cases that the implanted tissue engineering bone completely replaced the bone defect in 6th month displayed on X-ray,6 cases in 10th month,9 cases in 12th month and 1 case in 18th month. None of the cases had bone infection or bone absorption. Osteanagenesis and bone molding had come true in all cases. Conclusions The uncellular tissue engineering complex of autolegous red bone marrow wrapped by facial flap with vessels shows double effects in inducing osteanagenesis and the vascularization, and it is feasible in the recovery of large area bone defect. The complex can promote bone recovery and advance the quality and quantity of osteanagenesis.

Mice ; Animals ; Acetylation ; NF-kappa B/metabolism* ; Histone Deacetylases/metabolism* ; Inflammation ; Histone Deacetylase Inhibitors

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

¹⁷⁷Lu- prostate specific membrane antigen (PSMA) radio-ligand therapy has been approved abroad for advanced prostate cancer and has been in several clinical trials in China. Based on domestic clinical practice and experimental data and referred to international experience and viewpoints, the expert group forms a consensus on the clinical application of ¹⁷⁷Lu-PSMA radio-ligand therapy in prostate cancer to guide clinical practice.