AIM: To obtain the glycopohorin A (GPA) cDNA and construct the target gene in yeast two-hybrid.METHODS: About 410 bp cDNA fragment was amplified from K562 cell by RT-PCR.After being sequenced, the GPA gene fragment was cloned into EcoR -Ⅰ- Pst Ⅰ site of pbridge to form BD ends in yeast two-hybrid system. The recombinant plasmid was transfered into yeast AH109, and the expression in the yeast was also examined. RESULTS: The amino acid sequence encoded by cloned cDNA was mostly the same as reported GPA, and about 1 mm white yeast clone grew in the selective medium after 3 d.CONCLUSION: pbridge-GPA has nontoxic to the yeast, which can serve as a target gene in yeast two-hybrid system.

AIM: To study the effect of the overexpression of p16 on an anion exchange function of band 3 in HeLa cells. METHODS: The expression of p16 and band 3 in HeLa cells was detected by immunohistochemistry (IHC). The p16 cDNA was subcloned to plasmids pEGFP-C1 by PCR and identified by restriction enzyme digestion and sequencing, and then, the recombinant pEGFP-C1-p16 plasmids were transiently transfected into HeLa cells. The expression of fusion protein in HeLa cells was detected by fluorescence microscope. 6-methoxy-N-(3-sulfopropyl)-quinolinium(SPQ)fluorescent probes were used to detect the anion exchange function of band 3. RESULTS: P16 and band 3 were expressed in HeLa cells. The amplificated p16 cDNA sequence was the same as the report sequence. The transfective efficacy of pEGFP-C1-p16 was above 60%. The anion exchange function increased after the transfection of pEGFP-C1-p16 plasmids. CONCLUSION: p16 facilitates the anion exchange function of band 3 in HeLa cells.

OBJECTIVE: To purify hVEGF_(121)/EGFP fusion protein using transfected BMSCs as culture media, in addition, to detect the function of hVEGF_(121)/EGFP fusion protein in vitro.METHODS: The pEGFP-N_2-hVEGF_(121) recombinant plasmid, which was constructed in the preliminary work of our study group,was used to extract the plasmid DNA. BMSCs were transfected with pEGFP-N2-hVEGF_(121) by positive ionic liposome transfection method. Under a fluorescent microscopy, the expression of hVEGF_(121)/EGFP fusion protein was detected. The hVEGF_(121)/EGFP fusion protein was purified with Am icon ultrafiltration centrifuge tube and the expression of fusion protein was detected by Western-Blotting method.RESULTS: The BMSCs, which transfected with pEGFP-N2-hVEGF_(121), was observed under the fluorescent microscope. Western blotting confirmed that pEGFP-N_2-hVEGF_(121) fusion protein expressed in the culture media of transfected BMCS. MTT results showed the number of human umbilical vein endothelial cells in the fusion protein team was significantly greater than that in the control group (P < 0.05), and Miles test confirmed that pEGFP-N_2-hVEGF_(121) fusion protein increased the permeability of the blood vessel wall.CONCLUSION: ①This study successfully confirmed the pEGFP-N_2-hVEGF_(121) recombinant plasmid, which carrying VEGF_(121)/EGFP fusion protein, can be expressed in BMSCs.②The VEGF_(121)/EGFP fusion protein have the function of wild-type VEGF in vitro.

AIM: To investigate whether or not short-term high pressur e proliferates human umbilical vein endothelial cells (HUVECs), and the role of pp60c-src in cell proliferation. METHODS: Cultured HUVECs of 3-6th passa ge were exposed to atmosphere 0 mmHg (AP), 120 mmHg (MP), 180 mmHg (HP) and interfered with cap top ril(Cap, 10 ?mol/L)or irbesartan(Irb, 10 ?mol/L) Cell proliferation was q uantified by measuring hexosaminidase activity. The content of pp 60 c-src in cells was investigated by Western blotting. RESULTS:① Hexos aminidase activities which reflected cell number increased significantly at 4 h in MP and HP(0 145?0 018 and 0 144?0 019 vs 0 118?0 003,P

AIM: The experiment was designed to study the effect of compound Danshen dripping pills (DSDP) on myocardium with anoxin/reoxygenation. METHODS: The myocardial anoxin/reoxygenation model was made in perfused isolated rat heart. DSDP and isosorbide dinitrate (ID) were given at the time of pre-perfusion and reperfusion, then HPLC and H-600 electron microscope were used to detect the change of high energy phosphate and the ultrastructure of myocardial cell. RESULTS: ① The contents of AMP, ADP, ATP and AN in myocardium in only anoxin/reoxygenation group were obviously lower than those in the control group (P0.05). ③ In the groups with ID, the contents of AMP, ADP, ATP and AN were distinctively lower than those in the control group (P

Objective: To explore the correlation of monocyte to HDL-C ratio (MHR) and post-operative slow lfow or no relfow in patients with ST-segment elevation myocardial infarction (STEMI) after percutaneous coronary intervention (PCI). Methods: A total of 216 STEMI patients treated in our hospital from 2014-10 to 2016-05 were enrolled. The patients were divided into 2 groups: Slow lfow or no relfow group, the patients with TIMI grade≤2,n=43 and Normal lfow group, n=173. Receiver operating characteristic (ROC) curve was performed to assess the best cut-off value for MHR predicting slow lfow or no relfow with its sensitivity and speciifcity; Logistic regression analysis was conducted to studied weather MHR could be used as an independent risk factor for coronary slow lfow or no relfow in STEMI patients after PCI. Results: Compared with Normal lfow group, Slow lfow or no relfow group had the higher MHR (18.6±9.8) vs (10.9±5.5), P<0.001. Univariate Regression analysis indicated that MHR was a risk factor of slow lfow or no relfow occurrence (OR=2.22, 95% CI 1.58-3.28); multivariate regression analysis presented that MHR was an independent risk factor of slow lfow or no relfow occurrence (OR=1.55, 95% CI 1.01-2.38). ROC curve showed that the best cut-off value for MHR predicting slow lfow or no relfow occurrence was 13.37 with the sensitivity and speciifcity at 67.4% and 70.5% respectively, the area under curve (AUC) was 0.734, 95% CI 0.646-0.822. Conclusion: MHR was an independent risk factor for slow lfow or no relfow occurrence in STEMI patients after PCI.

In this study, about 350 bp cDNA fragment was amplified by PCR. After being sequenced, the AE1-c-end gene fragment was cloned into EcoR I-Pst I site of pGADT7 to form AD ends in the yeast two-hybrid system. The recombinant plasmid was transformed into yeast AH109, and the expression in the yeast was observed. The results demonstrate that AE1-c-end was obtained. pGADT7-AE1-c-end has no toxic effect on the yeast. It can serve as a target gene of yeast two-hybrid system.
Cloning, Molecular ; DNA, Complementary ; Plasmids ; Polymerase Chain Reaction ; Two-Hybrid System Techniques ; Yeasts

Objective:To observe the analgesic effect of local anesthesia combined with nerve block anesthesia on golden microneedles for improving facial aging.Methods:Between December 2018 and December 2019 in Burn and Plastic Surgery of Nanchong Central Hospital, sixty female patients (between 30 and 58 years old, with an average of 45.2 years old) with natural facial skin aging were randomly divided into two groups: Group A: surface anesthesia group (30 cases); Group B: local anesthesia combined with nerve block anesthesia (30 cases). Intraoperative and postoperative pain scores, length of operation, and incidence of adverse reactions were compared between groups A and B.Results:Pain score during surgery was (6.90±0.96) points in Group A, (3.63±0.72) points in Group B. The difference between the two groups was statistically significant ( t=14.93, P<0.05); Pain score at 30 minutes after operation was (2.03±0.62) in Group A, (0.77±0.73) in Group B, the difference between the two groups was statistically significant ( t=7.28, P<0.05). There was no statistically significant difference between the two groups in the pain score at 24 hours after operation ( P>0.05); The operation process in group B was simplified, and the treatment time was significantly shortened. The difference between the two groups was statistically significant ( t=17.93, P<0.05). Conclusions:The method of local anesthesia combined with nerve block anesthesia is used in the treatment of gold microneedles to improve the analgesic effect in facial aging, which significantly shortens the treatment time and has fewer adverse reactions. This method is worth popularizing.

Objective To evaluate the effect of multi?plate reconstruction plate in the treatment of partial complex tarsometatarsal joint injury injuries. Methods Seven patients treated complex tarsometatarsal joint injuries with multi?plate reconstruction plate in Nanjing Hospital Affiliated to Nanjing Medical University from September 2014 to July 2016 were selected in this study. According to Myerson classification,3 cases were A type,3 cases were B type,1 case was C type. The therapeutic effects were observed. Results The patients were followed up for 6-12 months,with an average of (8. 6±2. 0) months. According to the foot scoring criteria in American Orthopedic Foot and Ankle Society ( AOFAS) ,the function of the foot was evaluated,3 cases were in excellent condition,4 cases were in good condition. Conclusion In the case of multiple metatarsal fractures of the metatarsal base involved in the joint surface, the use of multiple reconstructive plates for joint fixation reduces the iatrogenic damage of the joint surface. The fixation effect and the functional recovery are satisfactory

Objective:To investigate the therapeutic effect of skull drilling and/or grinding combined with artificial dermis and vacuum sealing drainage in repairing scalp defects with skull exposure.Methods:From October 2014 to May 2018, 18 patients with scalp defect and skull exposure were treated in the Department of Burn and Plastic Surgery, the Second Clinical College of North Sichuan Medical College, including 10 males and 8 females, with an average age of 64 years (range, 34-86 years). The patients were divided into two groups: group A (by drilling skull or/and grinding combined with artificial dermis cover and vacuum sealing drainage plus two split thickness skin graft repair) and group B (by drilling skull or/andgrinding combined with artificial dermis cover plus two covering leather grinding stage split thickness skin graft repair), 9 cases in each group. The head wound granulation tissue, postoperative complications, skin graft survival rate and wound healing time were compared between the two groups. Vancouver scar assessment scale (VSS) was used to evaluate the wound healing in the two groups.Results:The time of granulation cultivation in group A and group B was (16.44±1.42) days and (29.11±13.32) days, the difference was statistically significant ( P<0.05); The wound healing time of group A and group B was (26.00±3.32) days and (40.67±14.37) days, the difference was statistically significant ( P<0.05); The postoperative complications of group A and group B were 1 case and 5 cases respectively, the difference was statistically significant ( P<0.05). The skin graft survival rates of group A and group B were (97.11±3.44)% and (95.00±4.74)%, the difference was not statistically significant ( P>0.05); The wound scar VSS scores of group A and group B were (7.67±1.32) points and (8.78±1.99) points, the difference was not statistically significant ( P>0.05). Conclusions:By drilling skull and/or grinding combined with artificial dermis cover and vacuum sealing drainage and two stage split thickness skin graft for repairing scalp defect with skull exposure wound can not only better scalp defect with skull exposure wounds, and reduce the postoperative complications, and significantly accelerate wound healing, but also can effectively improve the quality of wound healing, which is worthy of clinical application.