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@#Objective To summarize the clinical result of a combined technical system for bicuspid aortic valve (BAV) repair. Methods Patients who diagnosed as BAV and sever aortic regurgitation (AR) underwent a strategy of combined repair technics including annuloplasty, sinus plasty, leaflet plasty, sinus-tubular junction (STJ) plasty depending on anatomy pathological characteristics between October 2019 and January 2021 were enrolled. The clinical data of the patients were analyzed. Results A total of 17 patients were enrolled. There were 11 males and 6 females with an average age of 18-49 (32.4±13.6) years. Fifteen patients had typeⅠand 2 patients had typeⅡBAV according to Sievers classification. Annuloplasty was applicated in 13 patients, sinus plasty in 8 patients, leaflet plasty in 17 patients, and STJ plasty in 11 patients, respectively. The cardiopulmonary bypass (CPB) time was 95 (84, 135) min, aortic cross-clamping time was 68 (57, 112) min, and the ICU stay time was 17 (12, 25) h. After the operation, mild AR was presented in 14 patients, moderate AR in 1 patient and severe AR in 2 patients. The latter 3 patients underwent second operation under CPB, after then, 1 patient had mild AR and 2 patients had moderate AR. The follow-up time was 13.1±4.6 months. At the latest follow-up, 12 patients had mild AR and 5 patients had moderate AR, and no patient had reoperation. Conclusion A combined technical system for BAV repair can be used effectively and safely with an acceptable short and middle-term result.
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Tumor-targeted immunotherapy is a remarkable breakthrough, offering the inimitable advantage of specific tumoricidal effects with reduced immune-associated cytotoxicity. However, existing platforms suffer from low efficacy, inability to induce strong immunogenic cell death (ICD), and restrained capacity of transforming immune-deserted tumors into immune-cultivated ones. Here, an innovative platform, perfluorooctyl bromide (PFOB) nanoemulsions holding MnO2 nanoparticles (MBP), was developed to orchestrate cancer immunotherapy, serving as a theranostic nanoagent for MRI/CT dual-modality imaging and advanced ICD. By simultaneously depleting the GSH and eliciting the ICD effect via high-intensity focused ultrasound (HIFU) therapy, the MBP nanomedicine can regulate the tumor immune microenvironment by inducing maturation of dendritic cells (DCs) and facilitating the activation of CD8+ and CD4+ T cells. The synergistic GSH depletion and HIFU ablation also amplify the inhibition of tumor growth and lung metastasis. Together, these findings inaugurate a new strategy of tumor-targeted immunotherapy, realizing a novel therapeutics paradigm with great clinical significance.
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Objective:To summarize the construction of a technics system of aortic valve repair in adult patients with aortic regurgitation(AR).Methods:From March 2013 to May 2020, aortic valve repair was performed in 79 patients. In which 43 cases underwent simple technics such as subcommissural annuloplasty, leaflet plication etc. Cases done before May, 2018 were defined as A group, yet the other 36 cases underwent a routine repair technics system including annuloplasty, sinusplasty, cuspidplasty and sinotubularplasty after then as B group. The general data and the operation procedure, also the follow-up were analyzed.Results:In group A, there were 2 cases with simple aortic valve repair and 41 cases with other intracardiac operations. In group B, 23 patients had aortic valve repair alone and 13 patients had other intracardiac operations. Group A mostly adopts the technologies of subjunctional involution and ridge suspension. In group B, flap ring molding, flap leaf molding, sinus tube joint molding and other technologies were used. Postoperative reflux was reduced to moderate or mild in most patients in group A. In group B, postoperative reflux was reduced to mild in most patients. There were 7 cases of severe reflux in group A, and 2 cases of severe reflux were worse after repair than before surgery. In group B, 8 patients had no effective repair, and 3 patients continued to have satisfactory repair after the second turnaround. Aortic regurgitation at 1 year(or the last echocardiographic follow-up) after surgery: 15 patients in group A were mild, 17 were moderate, and 4 were severe; In group B, 2 patients underwent revalve replacement within 1 year, and the remaining 29 patients were moderate and mild, 20 cases were moderate, 7 cases were moderate, and 2 cases were severe.Conclusion:In some adult moderate and severe AR cases, a routine combined repair system can be applied to increase success rate of repair, and shows a good short and mid-term result.
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@#Objective To explore the effect of whether or not to stop beating after conversion to cardio-pulmonary bypass (CPB) in off-pump coronary artery bypass grafting. Methods From 2016 to 2018, 177 patients with off-pump coronary artery bypass grafting in Beijing Anzhen Hospital were transferred to CPB. According to whether they stopped beating after conversion to CPB during the operation, they were divided into two groups. A non-stop beating group: there were 76 patients with 45 males, 31 females. aged 63.53±6.98 years, who were not to stop beating after conversion to CPB. A stop beating group: there were 101 patients with 66 males and 35 females, aged 63.98 ± 8.37 years, who were to stop beating and underwent the modified perfusion and application of papaverine in perfusion after conversion to CPB. The clinical effect of the two groups was compared. Results There were 14 deaths in the perioperative period. The mean graft flow (MGF) in the stop beating group was higher (P=0.033), and the pulse index (PI) was lower (P=0.001) than those in the non-stop beating group. Intra-aortic balloon counter pulsation (P=0.036), extracorporeal membrane oxygenation (P=0.038), continuous renal replacement therapy (P=0.014), ventilator-assisted time (P=0.021), ICU monitoring time (P=0.012), perioperative mortality (P=0.025) and the ejcetion fraction value (P=0.023) were significantly different between the groups. Conclusion Compared with not to stop beating, those to stop beating can get better perioperative clinical effect after conversion to CPB, which is worthy of recommendation.
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@#Objective To investigate the influence of different discontinuation time of clopidogrel and aspirin before off-pump coronary artery bypass grafting on postoperative volume of drainage and blood products imported. Methods A total of 454 patients who underwent coronary artery bypass grafting in Beijing Anzhen Hospital from January 2017 through December 2019 were included. According to the preoperative discontinuation of clopidogrel and aspirin, all the 454 patients were divided into three groups including a guide group, a non-stop group and a stop group. There were 86 patients in the guide group including 59 males and 27 females with an average age of 64.12±6.15 years. They continued to take aspirin 100 mg/d before operation, but stopped clopidogrel for more than 5 days. In the non-stop group, there were 234 patients including 141 males and 93 females with an average age of 63.71±7.01 years. They continued to take aspirin 100 mg/d before operation, and stopped clopidogrel <5 days. In the stop group, there were 134 patients including 76 males and 58 females with an average age of 62.90±7.78 years. They stopped aspirin and clopidogrel for more than 5 days before operation. The clinical effectiveness was compared among the three groups. Results No perioperative death occurred in all patients. There was no statistical difference in platelet count, coagulation function, liver function, renal function, or myocardial markers among the groups (P>0.05). The hemoglobin [97 (15) g/ L vs. 98 (21) g/L vs. 100 (20) g/L, F=4.894, P=0.008] in the non-stop group was lower than that in the guide group and the non-stop group at 30 minutes postoperatively. The flow volume (399.87±127.19 mL vs. 367.05±125.89 mL vs. 349.63±130.68 mL, F=7.770, P=0.000) in the non-stop group at 3 hours postoperatively, the flow volume [600 (300) mL vs. 580 (245) mL vs. 550 (350) mL, Z=8.218, P=0.016] in the non-stop group at 6 hours postoperatively, the flow volume [750 (370) mL vs. 730 (350) mL vs. 730 (350) mL, Z=8.329, P=0.016] in the non-stop group at 12 hours postoperatively, the flow volume [890 (365) mL vs. 850 (340) mL vs. 850 (350) mL vs. Z=6.585, P=0.037] in the non-stop group at 24 hours postoperatively and the flow volume [950 (375) mL vs. 940 (360) mL vs. 940 (380) mL, Z=8.680, P=0.013] in the non-stop group at 48 hours postoperatively were more than those of the guide group and the stop group. The retention time of drainage tube was longer in the non-stop group [3 (1) d vs. 3 (1) d vs. 3 (1) d, Z=6.579, P=0.037] than in the guide group and the non-stop group. The amount of suspended erythrocytes input [0 (2) U vs. 0 (2) U vs. 0 (0) U, Z=6.150, P=0.046], and the amount of plasma input [200 (200) mL vs. 0 (200) mL vs. 0 (200) mL, F=4.144, P=0.016], the number of cases of plasma input (119 patients vs. 34 patients vs. 47 patients, Z=10.116, P=0.006) were more than those of the guide group and the stop group. Conclusion Aspirin maintenance is recommended for patients before off-pump coronary artery bypass grafting. If not necessary, clopidogrel is discontinued for at least 5 days.
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@#Objective To explore the difference of myocardial injury between off-pump coronary artery bypass grafting (OPCAB) and modified perfusion on-pump coronary artery bypass grafting (ONCAB). Methods A total of 558 patients who underwent coronary artery bypass grafting in Beijing Anzhen Hospital from 2017 to 2019 were included. According to whether or not they received modified perfusion cardiopulmonary bypass, all the 558 patients were divided into two groups including an OPCAB group (OP group) and an ONCAB group (ON group). There were 465 patients in the OP group including 282 males and 183 females with an average age of 63.58±7.87 years. In the ON group, there were 93 patients including 64 males and 29 females with an average age of 63.91±7.51 years. Creatine kinase MB (CK-MB) and cardiac specific troponin I (cTnI) were measured 24 hours before operation, 30 minutes after operation, 12 hours after operation, 36 hours after operation and 48 hours after operation. Results No perioperative death occurred in all patients. CK-MB (5.00 ng/mL vs. 8.60 ng/mL, Z=–2.189, P=0.029) and cTnI (3.00 ng/mL vs. 7.80 ng/mL, Z=–5.307, P=0.000) in postoperative 12 hours in the ON group were less than those in the OP group. CK-MB (5.00 ng/mL vs. 5.60 ng/mL, Z=–2.280, P=0.023) and cTnI (0.10 ng/mL vs. 1.02 ng/mL, Z=–6.418, P=0.000) in postoperative 36 hours in the ON group were less than those in the OP group. cTnI (0.07 ng/mL vs. 0.81 ng/mL, Z=–1.946, P=0.032) in postoperative 48 hours in the ON group was less than that in the OP group. Conclusion Compared with OPCAB, modified perfusion ONCAB has less myocardial damage.
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@#Objective To analyze the protective mechanism of spinal cord ischemia-reperfusion injury mediated by N-methyl-D-aspartate (NMDA) receptor. Methods A total of 42 SD rats were randomly assigned to 4 groups: a non-blocking group (n=6), a saline group (n=12), a NMDA receptor blocker K-1024 (25 mg/kg) group (n=12) and a voltage-gated Ca2+ channel blocker nimodipine (0.5 mg/kg) group (n=12). The medications were injected intraperitoneally 30 min before ischemia. The neural function was evaluated. The neuronal histologic change of spinal cord lumbar region, the release of neurotransmitter amino acids and expression of spinal cord neuronal nitric oxide synthase (nNOS) were compared. Results At 8 h after reperfusion, the behavioral score of the K-1024 group was 2.00±0.00 points, which was statistically different from those of the saline group (5.83±0.41 points) and the nimodipine group (5.00±1.00 points, P<0.05). Compared with the saline group and nimodipine group, K-1024 group had more normal motor neurons (P<0.05). There was no significant difference in glutamic acid concentration in each group at 10 min after ischemia (P=0.731). The nNOS protein expression in the K-1024 group was significantly down-regulated compared with the saline group (P<0.01). After 8 h of reperfusion, the expression of nNOS protein in the K-1024 group was significantly up-regulated compared with the saline group (P<0.05). Conclusion K-1024 plays a protective role in spinal cord ischemia by inhibiting NMDA receptor and down-regulating nNOS protein expression; during the reperfusion, K-1024 has a satisfactory protective effect on spinal cord function, structure and biological activity of nerve cells.
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Objective To investigate the changes on apparent diffusion coefficient(ADC)of diffusion weighted imaging with dif-ferent b values and to assess the diagnostic value of ADC for discriminating malignant from benign orbital masses.Methods 81 pa-tients (55 benign orbital masses and 26 malignant orbital masses)were enrolled.These patients underwent serial MR and echo-pla-nar DW imaging examinations of the orbits with b values=0 and/or 400,700,1 000 s/mm2 .The differences of ADC values under different b values between benign and malignant orbital masses were observed and compared,and diagnosis of ADC values was as-sessed by the receiver operating characteristic (ROC)curve.Results In three groups,the ADC values of benign orbital masses [(1.56±0.35)×10 -3 mm2/s,(1.40±0.34)×10 -3 mm2/s and (1.30±0.32)×10 -3 mm2/s,respectively]were significantly differ-ent (P <0.01),while the ADC values of malignant orbital masses were not statistically different (P =0.093)[(1.12 ±0.47)× 10 -3 mm2/s,(0.92±0.42)×10 -3 mm2/s and (0.87±0.40)× 10 -3 mm2/s,respectively].The areas under the ROC curve of ADC values were 0.799,0.866 and 0.867,respectively.The threshold values were 1.05 × 10 -3 mm2/s,0.94 × 10 -3 mm2/s and 0.93 × 10 -3 mm2/s (while sensitivity were 65%,81% and 81%;specificity were 94.5%,91% and 85%;positive predictive values were 85%,80%,72%,negative predictive values were 85%,91%,90%,and accuracy were 85%,87% and 84%,respectively).Conclusion The b value of 700 s/mm2 offered better diagnostic performance.ADC values of diffusion weighted imaging can reflect diffusion characteristics of the orbital masses,as a complementary tool in the differentiation of malignant from benign orbital masses.
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OBJECTIVE@#To analyze the MR imaging findins of acinic cell carcinoma(ACC) in parotid gland and develop the diagnosis of this disease.@*METHOD@#The MR imaging fearures of 11 patients with pathologically proved ACC were retrospectively analyzed. MR imagings were analyzed in relation to the follow:amount, location, size, shape, margin, pseudocapsule, MR signal intensity and enhanced pattern and the cervical lymphadenopathy. The clinical analysis on patients included age, sex, and follow-up.@*RESULT@#There were 11 patients, 7 lesions was located in the right parotid gland and 4 lesions was located in the left parotid gland; the average maximum diameter was (2.66±0.99)cm; 7 lesions showed lobulated,and 4 lesions showed round; 8 lesions had no pseudocapsules and 3 lesions had incomplete pseudocapsule. All lesions showed homogeneous or heterogeneous isointense and slight hyperintense on T1 WI and T2WI. On post contrast images, the tumor parenchyma ingredients showed remarkable enhancement in all lesions.@*CONCLUSION@#The MR imaging of ACC in the parotid gland don't have specific features, but when the tumor of the parotid gland showed no pseudocapsule (or showed incomplete pseudocapsule), lobulate, small cysitc and remarkable enhancement, it may indicate ACC, and the correct diagnosis depends on clinical pathology.
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Humans , Carcinoma, Acinar Cell , Pathology , Contrast Media , Magnetic Resonance Imaging , Parotid Gland , Retrospective Studies , Salivary Gland Neoplasms , PathologyABSTRACT
Objective To investigate the role of PT H (1-34 ) on the expression of receptor activator factor of NF-κB ligand (RANKL) in the induction of osteoclasts and its effect to osteoclasts on compression side during the repairs of tooth root in rats model of tooth resorption by intermittent injection of small dose of PTH (1-34) .Methods After the model of tooth resorption was established in rats 6-8 weeks in age ,63 male SD rats were divided in three groups .Rats in the control group were not given injec-tions for any drugs ;The negative control group were given injections for normal saline 6μg/kg subcutaneously every other day ;The experimental group were given injections for PTH (1-34) 6 μg/kg(PTH :1 μg/mL) subcutaneously every other day ;then rats in every group were killed on the day 0 ,7 ,10 ,14 ,17 ,21 ,25 .TRAP staining for counting TRAP-positive stained osteoclasts ,calculat-ing the mean ;Ligand RANKL immunohistochemistry and using image-pro-plus image analysis system to measure the average opti-cal density value of compression side .Results On the day of discontinuation ,the tooth resorption continued in each group ;the num-ber of osteoclasts between every two arrays there were no significant statistic differences (P>0 .05);RANKL immunohistochemis-try :Compared with control group and the negative control group ,the experimental group significantly increased in early stage ,and reduced in latest stage(P<0 .05) .Conclusion It indicated that intermittent injection of small dose PTH (1-34) did not cease the rats tooth resorption which occured during the period of tooth repair .
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Purpose To compare the CT features of hepatic angiomyolipoma (HAML) and primary hepatocellular carcinoma (HCC) and improve diagnostic accuracy for HAML. Materials and Methods The CT findings of 9 patients with HAML confirmed pathologically and 20 patients with HCC were retrospectively analyzed in terms of lesion location, appearance, size, margin, CT value at each phase, cirrhosis and vessel presence inside tumor. Results Mean CT value on plain scan and at arterial phase on contrast enhanced scan of HAML was significantly lower than that of HCC (t=-2.885, P0.05). CT value on plain scan and at arterial phase of both HAML and HCC was not significantly different (t=1.289, P>0.05) whilst that at portal vein phase was significantly different (t=2.516, P<0.05). Central vessels were shown in 7 cases of HAML and 3 cases of HCC, pseudocapsule appeared in 1 case of HAML and 13 cases of HCC with statistical difference (χ2=10.828, 7.219;P<0.01). Conclusion CT scan of HAML reveals the component of fat and presentsfast in and slow outpattern, large shadow of vessel and absence of capsule on contrast enhanced scan, which can be helpful in the differential diagnosis from HCCs.
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Objective To investigate the diagnostic value of apparent diffusion coefficient (ADC)for discriminating orbital lymphomas from other orbital mass lesions.Methods In this prospective study,87 subjects (22 orbital lymphomas and 65 other orbital mass lesions) were enrolled.These patients underwent serial MR and echo-planar DW imaging examination of the orbits with b values of 0 and 700 s/mm2 at 1.5 T MR (GE Signa Excite).ADC maps were reconstructed,and the ADC values of the orbital masses were calculated.Mass ADC was also compared with that of normal-appearing white matter (ADC ratio,ADCR).The receiver operating characteristic curves (ROC) were constructed using optimal cut point of ADC and ADCR to differentiate between orbital lymphomas and other orbital mass lesions.The areas under the ROC curve for ADC and ADCR were also calculated.Results The mean ADC and ADCR of orbital lymphomas were (0.77 ± 0.17) × 10-3 mm2/s and 0.89 ± 0.21,respectively.The mean ADC and ADCR of other orbital mass lesions were (1.36 ± 0.38) × 10-3 mm2/s and 1.51 ± 0.43,respectively.Lymphomas had lower ADCs and ADCRs than other orbital mass lesions (t =-9.620,-9.003,P =0.000).The areas under the ROC curves of ADC and ADCR diagnosing lymphoma were 0.94±0.03 and 0.91 ±0.03,respectively.An ADC of less than 0.804 × 10-3 mm2/sec and ADCR of less than O.956 were optimal for predicting lymphoma (sensitivity,77.3％ for both; specificity,98.4％ and 92.3 ％,respectively ; and accuracy,93.1％ and 88.5 ％,respectively).Compared with pathological results,both ADC and ADCR had high correlations (Kappa values were 0.806 and 0.696,respectively).Conclusion Diffusion-weighted imaging can be applied as a complementary tool in the detection of orbital lymphomas.
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Objective: This study aimed to compare and analyze the functional differences between peripheral blood mono-cyte-derived dendritic cells (DCs) of Helicobacter pylori-positive and H. pylori-negative patients with gastric cancer. Methods:H. py-lori infection was detected in 84 patients with gastric cancer in our hospital from January 2011 to October 2012 by the 14C-urea breath test. DCs were generated from monocytes isolated by an adherent method from the two groups of patients and cultured in the presence of rhIL-4, rhGM-CSF, and rhTNF-α. Furthermore, the expression of surface marker molecules was determined by fluorescence-activat-ed cell sorting analysis. The cytotoxicity of DCs pulsed T cells against gastric carcinoma cell was assessed by the lactate dehydroge-nase-releasing assay. The secretion of IL-12 and IFN-γin the supernatant was determined by enzyme-linked immunosorbent assay. Re-sults:No difference was observed in the morphological change of the maturation process. The mean expression of CD1a, CD80, CD83, CD86, and HLA-DR molecules in DCs of H. pylori-infected patients was higher than that in DCs of H. pylori-negative group, and the differences were statistically significant except for CD1a and HLA-DR. The cytotoxicity activities, IL-12 release, and IFN-γrelease in the H. pylori-positive group were significantly higher than those in the H. pylori-negative group (P<0.05). Conclusion:H. pylori infec-tion has no effect on the morphological change of the maturation process of monocyte-derived DCs. These data clearly demonstrate that monocyte-derived DCs of H. pylori-infected patients with gastric cancer can induce stronger maturation and activation than those of H. pylori-negative patients.
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To investigate the influence of bear bile on rat hepatocarcinoma induced by diethylnitrosamine (DEN), a total of 40 rats were randomly divided into 4 groups: normal control group, model group, and two bear bile treatment groups. The rat liver cancer model was induced by breeding with water containing 100 mg x L(-1) DEN for 14 weeks. The rats of the bear bile groups received bear bile powder (200 or 400 mg x kg(-1)) orally 5 times per week for 18 weeks. The general condition and the body weight of rats were examined every day. After 18 weeks the activities of serum alanine transaminase (ALT), aspartate transaminase (AST) and total bilirubin (TBIL) were detected. Meanwhile, the pathological changes of liver tissues were observed after H&E staining. The expression of proliferative cell nuclear antigen (PCNA) and a-smooth muscle actin (alpha-SMA) in liver tissue were detected by immunohistochemical method. After 4 weeks the body weights of rats in normal group were significantly more than that in other groups (P < 0.05); and that in the two bile groups was significantly more than that in the model group. Compared with normal group, the level of serum glutamic-pyruvic transaminase and total bilirubin increased significantly in other groups; compared with model group, these two indexes decreased significantly in two bile groups. Hepatocellular carcinoma occurred in all rats except for normal group; there were classic cirrhosis and cancer in model group while there were mild cirrhosis and high differentiation in two bile groups. There were almost no expressions of PCNA and alpha-SMA in normal group while there were high expressions in model group; the two bile groups had some expressions but were inferior to the model group, and alpha-SMA reduced markedly. It indicated that bear bile restrained the development of liver cancer during DEN inducing rat hepatocarcinoma, which may be related to its depressing hepatic stellate cell activation and relieving hepatic lesion and cirrhosis.
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Objective To evaluate the quality of life status in patients aged 70 years and over following off-pump coronary artery bypass (OPCAB) grafting. Methods Seventy-eight patients with coronary heart disease [mean age (74.6 ± 5.3) years, 66 males, 12 females] were investigated retrospectively. Three questionnaires about the quality of life, including Seattle Angina Questionnaire (SAQ), Nottingham Healthy Profile (Part Ⅰ NHP) and Duke Activity Status Index (DASD, were used to investigate patients before and after OPCAB. Results Prior to OPCAB, there was lower quality of life index in males than in females [SAQ: (65.3±5.1) vs. (69.5±8.1); NHP: (89.4±17.3) vs. (125.2±19.9), P＜0.01; DASI: (4. 1±1.1) vs. (4.3± 1.3)]. At the 12th months after OPCABG, there were significant improvements in all patients. The effects were less pronounced in females than in males [SAQ: (83.1 ±5.8) vs. (88.5±4.5), P＜0.05; NHP: (84.7± 11.7) vs.(91.4±13.7), P＜0.05; DASI: (4.7±1.4) vs. (5.4±1.1)]. Conclusions Our study shows that OPCAB improves quality of life in elderly patients with coronary heart disease. The benefits of OPCAB are even more pronounced in male patients.
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Objective To evaluate the therapeutic strategy and effects of surgical treatment for patients with closed trauma of heart valves. Method A total of 32 patients, 28 male and 4 female, aged from 21 to 48 years old with closed trauma of heart valves treated with surgical intervention from June 1987 to November 2008 were retrospectively studied. The duration from trauma to surgical treatment ranged from 18 days to 4 years. Results There was no peri-operative mortality within 30 days. Clinical follow-up for one month to 22 years showed cerebral hemorrhage in only one patient 27 months later and sudden death in one patient probably due to choke of the valve.Conclusions The mortality of patients with closed trauma of heart valves may be decreased by timely and correct diagnosis and treatment, and the outcomes may be improved.
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Purpose To prepare a novel fusion protein of CREKA and tTF as a universal carrier targeting to cancer,and to analyze its activities.Methods CREKA and tTF gene were acquired by PCR,and inserted into plasmid pET22b(+)to construct recombinant plasmid CREKA/tTF/pET22b(+),and the fusion gene was expressed in E.coli BL21.The fusion protein Wag purified through Nickel-affinity chromatography column.After purifying,the fusion protein was refold by subsequent dialysis.The activities of the fusion proteins were measured by coagulation timing and quantitative fluorescence test in vitro.Results The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was obtained.The fusion protein was highly expressed in E.coli BL21.The coagulation of the fusion protein Was determined by the coagulation test.And the capability of the fusion protein effectively binding to clotted plasma proteins is identified in quantitative fluorescence test.Conclusion The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was built.The fusion protein CREKA/tTF with both TF and CREKA activity was successfully obtained.
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Background and purpose: Tumor vasculature is increasingly recognized as a target for cancer therapy. In recent years, a fusion protein consisting of the extra cellular domain of tissue factor (truncated tissue factor, tTF) was fused to the antibody selectively binding to tumor vasculature. Antibody-truncated tissue factor(Ab-tTF) fusion protein specifically induced thrombotic occlusion of tumor vessels resulting in tumor growth retardation or regression in some types of solid tumors. However, there were still some disadvantages in the above approach. We constructed and expressed that the (RGD)_3-tTF fusion protein with peptides arginine-glycine-aspartic acid (GRGDSP, abbr. RGD)as the carrier of tTF to explore whether it bad the capability of targeting to tumor vasculature in the colonic carcinoma model. Methods: The (RGD)_3-tTF fusion gene consisting of the tTF was fused to three series-wound peptides RGD. The (RGD)_3-tTF construct was expressed in Escherichia coil BL21(DE_3). The fusion protein was purified through Nickel affinity chromatography column. The activity of inducing blood coagulation was detected by clotting assay and coagulation factor X (FX) activation assay. The specific binding to integrins α_vβ_3 was analyzed by indirect enzyme linked immunosorbent assay (ELISA). All these were compared with the fusion protein RGD-tTE Colonic nude mice models were randomly divided into 3 groups (1 nude mice per group).Tumors were stained by the (RGD)_3-tTE RGD-tTF fusion protein and tTF which were labeled with Fluorescein Isothiocyanate(FITC). The location of the (RGD)_3-tTF fusion protein in the colonic carcinoma bearing nude mice tissue was analyzed by immunofluorescence assay. Results: The (RGD)_3-tTF fusion protein retained tissue factor thrombogenic activities. With increasing concentration, the clotting time was shortened correspondingly. Under the conditions of Ca~(2+), the clotting time was 9.96±0.56 min when the concentration was 6 μmol/L(P<0.01). The (RGD)_3-tTF fusion protein could activise F X above 6 μmol/L concentration, which was similar to RGD-tTF fusion (F=0.147, P>0.05). The ability of the (RGD)_3-tTF fusion protein binding specifically to integrins α_vβ_3 was stronger than that of the RGD-tTF fusion protein in the same concentration (F=164.81, P<0.01), which was apparently indicated by the A_(405nm) 1.25 and 0.95 when the concentration was 0.24 μmol/L. Immunofluorescence assay showed that the (RGD)_3-tTF fusion protein was assembling in the tumor vasculature of the colonic carcinoma bearing nude mice. Conclusion: The (RGD)_3-tTF fusion protein which retained tissue factor thrombogenic activities could bind specifically and efficiently to tumor vasculature in the colonic carcinoma bearing mice through binding to the tumor marker integrins α_vβ_3. It might be a promising foundation for further studies on the colon cancer molecular targeted therapy with tTF as an effective factor.
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The amino acid sequence (1-301aa) coding the human PTP1B catalytic domain (PTP1Bc) was obtained from the GenBank. The PTP1Bc gene was constructed by overlapping PCR, then was inserted into vector pET-22b(+) and expressed efficiently in E. coli BL21(DE3) under optimum condition after IPTG induction. The proteins were expressed mainly as inclusion bodies with the yield of more than 30% of total bacterial proteins. The expressed products were purified through Ni(2+)-affinity chromatographic column. After purification, the purity of the proteins was more than 95%. Western blotting analysis suggested that the purified proteins could specially combine with anti-PTP1B antibody. Enzyme activity assay showed that the protein has phosphatase activities.
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Humans , Catalysis , Escherichia coli , Genetics , Metabolism , Genetic Vectors , Genetics , Inclusion Bodies , Metabolism , Polymerase Chain Reaction , Methods , Protein Tyrosine Phosphatase, Non-Receptor Type 1 , Genetics , Recombinant Proteins , Genetics , MetabolismABSTRACT
Objective To summarize the clinical experience of surgical treatment for the congenital coronary artery anomaly.Methods 15 patients with coronary artery anomaly,congenital coronary artery fis-tulas were 11 patients,including 1 patient associated with Fallot,5 patients underwent ligation of isolated fistula and 1 patient's fistula was performed tangent suture though off-pump cardiopulmonary bypass,4 pa-tients'fistulas were repaired by endocardiac way and 1 patient was mended through dissection of fistula under Oil-pump cardiopulmonary bypass.Anomalous origin of the left coronary artery from pulmonary artery were 4 patients and 1 patient associated with rheumatoid valve disease.Operative methods included ligation of left circumflex (1 case),anastomosis of left subclavian artery-left artery-left main (1 case) and transplantation of left main(2 cases).Results There was 1 death becaused severe low cardiac output syndrome,and the there was good prognosis and no clinical symptoms found in 14 patients during follow-up.Conclusion The feasible operation for the coronary artery anomaly should be performed as early as possible once diagnosed correctly.