Objective To obtain eukaryotic expression vectors containing coding region of nephroblastoma overexpression gene (NOV) and detect its expression in COS-7 cells. Methods A 1 165-bp cDNA fragment was amplified from the total RNA of normal rat brain tissue by RT-PCR and cloned into eukaryotic expression vector pcDNA3.1/Myc-His(+)/lacZ. The cloned insert was identified by double digestion of the recombinant plasmid with restriction enzymes HindⅢ and BamHⅠ. The recombinant plasmid was transfected into COS-7 cells with liposome. The expression of NOV gene was detected by Western blotting and immunocytochemistry. Results Eukaryotic expression vectors containing 1 165 -bp coding region of NOV gene was constructed. COS-7 cells transfected with the recombinant plasmid expressed high level of NOV protein in cytoplasm. Conclusion That eukaryotic expression vectors containing coding region of NOV gene was constructed can provide a strong molecular tool for the studies of effect of NOV gene.

Objective To analyze influence factors of serum A-TG level after DTC 131 I treatment ,to explore whether A-TG can be used as an indicator of follow-up ,recurrence and metastasis of DTC .Methods From 2008 January to 2013 February ,a total of 106 DTC patients underwent operation excisions of thyroid radioiodine were treated with 131 I .Before and 1 month after treatment , the levels of A-TG ,TG ,THS were measured .The relationship of A-TG and thyroid operation range operation times ,thyroid gland residual volume and time ,the levels of TSH and TG ,thyroid cancer metastasis and recurrence were examined .Results Serum A-TG concentration associated with operation scope ,frequency(P<0 .05) .It was showed that the A-TG concentration was positively correlated with the metastasis of thyroid cancer .Conclusion The serum TG level detection combined with A-TG and 131 I-WBS can improve the diagnostic sensitivity and accuracy of DTC recurrence and metastasis .

Objective To investigate the electrophysiological characteristics of atrial tachycardia (AT) originating from the side of the bundle of His, and to analyze the efficacy and safety of transcatheter radiofrequency ablation via the right side of the bundle of His and left noncoronary sinus of aorta for the treatment of AT. Methods A total of 12 patients with AT originating from the side the bundle of His, which was confirmed by conventional electrophysiological study and atrial activation patterns, were included in this study. The patient’s age ranged from 12 to 64 years old with a mean of (47.4±14.6) years. The mapping was carried out at the right side of the bundle of His and radiofrequency ablation was performed. When the ablation procedure failed, or the junction zone rhythm or atrioventricular block occurred, ablation via the left noncoronary sinus of aorta was employed. Results Atrial stimulation could repeatedly induce and terminate AT in all the 12 patients, the average cycle length was (327±76) ms. TA was terminated within 10 seconds after the start of ablation in 10 patients during their AT attacks. Ablation was unsuccessful in 2 patients. Ablation via the right side of His bundle was successfully accomplished in 2 patients, and the ablation via the left noncoronary sinus of aorta was successfully carried out in 8 patients. The average follow-up time was 1-6 years, and no recurrence of AT was observed. Conclusion Atrial tachycardia originating from the side of the bundle of His has certain electrophysiological characteristics, and transcatheter radiofrequency ablation is safe and effective for its treatment. Ablation via the left noncoronary sinus of aorta should be considered as a strategy of priority.

ObjectiveTo study electrophysiological characteristics and catheter ablation left-sided accessory pathways of Mahaim fiber.MethodsTwenty-eight patients underwent catheter ablation for Mabaim fiber and 3 patients was left-sided accessory pathways.All of them were male with 34,20,38 years old.The tachycardia history was1-5years. Electrophysiological characteristics,target sitesand electrocardiogram(ECG,12 leads) were recorded through catheter from coronary sinus(CS),high right atrium(HRA),His bundle and ablation.ResultsOne patient's ECG showed little preexcitation,the other two were normal. Atrial stimulation could be easily induced tachycardia.Their ECG showed wide QRS waves (right bundle branch block or similar right bundle branch block).The characteristics of accessory pathways were slow and anterograde decremental conduction as well as blocked by adenosine triphosphate injection.Ventricular-atrial conduction was via atrioventricular node with centripetal decreasing transmission during ventricular pacing.During tachycardia,the His bundle electrogram resulted in a V-H-A pattern,the retrograde A in His bundle was pioneer.The atriofascicular pathways of 2 patients were completely ablated respectively in left posterior lateral and before the free wall of mitral valve ring.The nodoventricular pathway of 1 patient was successfully ablated in left middle posterior lateral of mitral valve ring.ConclusionsThe left-sided accessory pathways of Mahaim fiber is an uncommon accessory pathway,which has specific electrophysiological characteristics.It is effective and safe to ablate accessory pathway with radiofrequency.

Objective:To explore the identification value of carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC-Ag), cytokeratin 19 fragment (Cyfra211), thyroglobulin (TG), ferritin (Fer) and procalcitonin (PCT) in fine needle aspiration eluent in benign and malignant cervical nodules, and acquire the optimal diagnostic model.Methods:Three hundred and ninety-six single cervical nodule patients who underwent fine needle aspiration biopsy from August 2017 to August 2019 in the Center Hospital of Xiaogan City of Hubei Province were selected. The fine needle aspiration eluent levels of CEA, SCC-Ag, Cyfra211, TG, Fer and PCT were detected by electrogenerated chemiluminescence method. The results of cytopathological diagnosis were regard as "gold standard", and the diagnostic efficiency of single and combined indexes in fine needle aspiration eluent were analyzed by receiver operating characteristic (ROC) curve.Results:Among the 396 patients, malignant nodules was in 101 cases, and benign nodules was in 295 cases. The fine needle aspiration eluent levels of CEA, SCC-Ag, Cyfra211, TG and Fer in patients with malignant nodules were significantly higher than those in patients with benign nodules: (27.73 ± 10.63) μg/L vs. (16.81 ± 8.18) μg/L, (1.59 ± 0.74) μg/L vs. (1.09 ± 0.83) μg/L, (3.31 ± 1.48) μg/L vs. (1.66 ± 0.59) μg/L, (144.96 ± 38.93) μg/L vs. (95.03 ± 47.23) μg/L and (191.18 ± 80.13) μg/L vs. (137.87 ± 63.22) μg/L, the PCT was significantly lower than that in patients with benign nodules: (0.61 ± 0.24) μg/L vs. (1.01 ± 0.52) μg/L, and there were statistical differences ( P<0.01). The ROC curve analysis result showed that the CEA, Cyfra211 and TG had super diagnostic value (area under curve>0.7, Youden index>0.5); the area under curve of CEA, Cyfra211 combined with TG was significantly higher than other combined detection of 2 indexes ( P<0.05). Conclusions:The combined detection of CEA, Cyfra211 and TG in fine needle aspiration eluent can effectively distinguish the benign and malignant cervical nodules.

Objective To investigate the risk factors that may induce pneumothorax resulted from subclavian vein puncture during interventional therapy for arrhythmia.Methods The clinical data of 4 351 patients with heart disease,who were admitted to Wuhan Asia Heart Disease Hospital,China,during the period from January 2010 to January 2014 to receive subclavian vein puncture for the performance of radiofrequency catheter ablation or pacemaker implantation,were retrospectively analyzed.The gender,age,presence of chronic obstructive pulmonary disease (COPD) and the shape of clavicle of patients who developed pneumothorax were analyzed,and their relationships with the occurrence of pneumothorax were evaluated.Results Of the 4 351 patients,47 patients developed pneumothorax(1.08％),their age was 12-72 years old with a mean of (47.1±18.4) years old.Among the 47 patients,37 patients were females (78.7％).The angle between clavicle and sternum ＜60°(oblique clavicle) was seen in 27 patients,coexisting COPD was seen in 2 patients.The occurrence of pneumothorax was strikingly higher in females than that in males (OR=2.7,95％ CI=1.4-5.2).In patients with oblique clavicle the risk of pneumothorax was obviously increased (OR=3.5,95％ CI=1.6-7.9) and in patients with COPD the probability of pneumothorax was remarkably increased (OR=2.3,95％CI=1.2-4.7).No special treatment was employed in 5 pneumothorax patients,and the pneumothorax was absorbed by itself.In 19 patients,the pneumothorax was relieved through thoracic puncture and suction.Closed drainage of thoracic cavity was employed in 23 patients.All patients were discharged from hospital with complete rehabilitation.Conclusion Clinically,pneumothorax has been an important clinical problem in interventional therapy.In female patients and in patients who have oblique clavicle or coexisting COPD,pneumothorax is more likely to occur when they receive subclavian vein puncture.

Objective To evaluate the clinical effect of nanofat on superficial rhytides of face and neck and dark lower eyelids.Methods From September 2014 to January 2017,a total of 86 cases were collected in our hospital for voluntary nanofat transplantation on superficial rhytides and dark lower eyelids.This was a retrospective study,which included 18 cases of eye wrinkles,14 cases of forehead wrinkles,neck wrinkles in 14 cases,and 22 cases of dark lower eyelids.First of all,we harvested mircofat through liposuction,and then transformed mircofat to nanofat.At last,nanofat was grafted into intradermal layer of the skin with sharp needles.We took the standard photographs of the patients.After six months follow-up,doctors and patients evaluated the short term and long-term postoperative effect.Results No serious complications occurred in all patients.Postoperative evaluation of facial and neck superficial wrinkles showed that the satisfactory rate of doctors and patients after one month,was 85.9％and 84.3％,and after 6 months 87.5％ and 84.3％,respectively.Postoperative evaluation of dark lower eyelids showed that the satisfactory rate of doctors and patients after one month was 45.5％ and 36.4％,and after 6 months 81.8％ and 86.4％,respectively.Conclusions Nanofat can rectify the superficial rhytides of face and neck and dark lower eyelids in some patients.However,for some patients the effect is not satisfied.

Objective:To investigate the auxiliary diagnostic value of serum midkine (MK) and thyroid stimulating hormone (TSH) for differentiated thyroid cancer (DTC).Methods:Seventy-one postoperative DTC patients (DTC group) treated with 131I were selected, and 143 patients with benign thyroid lesions (benign thyroid disease group) treated with surgery in Center Hospital of Xiaogan from March 2019 to December 2020 at the same period were also selected. Clinical data such as liver and kidney function indexes, positive rate of anti thyroglobulin antibodies (TGAb) and positive rate of thyroid peroxidase antibody (TPOAb) were collected before treatment, and their fasting blood samples were collected before treatment. Fully automated electrochemiluminescence immunoassay was used to measure free thyroxine (FT 4), free triiodothyronine (FT 3), TSH levels in patients′ serum. The serum MK levels were measured by enzyme-linked immunosorbent assay (ELISA). Binary Logistic regression model was used to screen for independent risk factors for the development of DTC. A receiver operating characteristic curve (ROC) was drawn to evaluate the efficacy of MK, TSH and MK combined with TSH, in aiding the diagnosis of DTC and its staging. Results:Serum TSH and MK levels in DTC group were higher than those in benign thyroid disease group: (3.55 ± 0.61) mU/L vs. (2.97 ± 0.46) mU/L, (394.25 ± 63.36) ng/L vs. (311.45 ± 42.66) ng/L, and the differences were statistically significant ( P<0.05). Elevated serum TSH and MK levels were independent risk factors for DTC. When MK combined with TSH was used to diagnose DTC, the area under the curve (AUC), sensitivity and specificity were higher than those of MK and TSH alone (0.925 vs. 0.859 and 0.783, 83.10% vs. 78.87% and 73.24%, 89.51% vs. 85.31% and 79.02%), and the differences were statistically significant ( P<0.05). Serum TSH and MK levels in stage Ⅲ and Ⅳ patients in DTC group were higher than those in stage Ⅰ and Ⅱ patients: (3.79 ± 0.65) mU/L vs. (3.42 ± 0.56) mU/L, (427.88 ± 52.73) ng/L vs. (311.45 ± 42.66) ng/L, and the differences were statistically significant ( P<0.05). The AUC, sensitivity and specificity of MK combined with TSH in the diagnosis of different stages of DTC were higher than those of MK and TSH alone (0.822 vs. 0.657 and 0.666, 73.90% vs. 56.52% and 56.52%, 83.33% vs. 77.08% and 79.17%), and the differences were statistically significant ( P<0.05). Conclusions:Serum TSH and MK levels are independent risk factors for the occurrence of DTC in patients, and the combination of them has certain auxiliary diagnostic value for the identification and staging of DTC.

Studies on lycopene synthesis in Escherichia coli were not only able to gain the strains with high yield and less by-products, but also able to test functions of genes or gene clusters. In this article, the cDNA sequences of tomato LeGGPS2 and LePSY1 as well as the coding sequence of crtI from Erwinia uredovora, each of which was added a ribosome biding site, were controlled by T7 promoter and terminator alone or combined, and expressed in E. coli BL21 (DE3) to induce lycopene synthesis. The results show that only T7::crtI-LeGGPS2-LePSY1 expressed tri-cistronically could produce lycopene, and 2.124 mg/g dry cell weight oflycopene was obtained when fermented for 5 h at 30 degrees C after mixing 80 micromol/L IPTG at the later logarithmic phase while the seed broth of 1:50 (V/V) was inoculated into LB medium (pH 6.8) containing 3% sucrose and cultured for 8 h at 37 degrees C. The results confirmed the function of the prokaryonized LeGGPS2 and LePSY1 and their synergy with crtI, and also laid a foundation to establish an independent lycopene synthetic pathway in the tomato plastid.
Bacterial Proteins ; genetics ; Carotenoids ; biosynthesis ; genetics ; Cloning, Molecular ; Erwinia ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Engineering ; Plant Proteins ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

Objective: To explore the nipple-areola complex blood supply mode in hypertrophic breasts, and to obtain the pertinent knowledge of vascular anatomy for breast reduction surgery as well as the analysis of similarities and differences between hypertrophic and normal breasts. Comparing the blood supply of nipples-areola complex between these two groups for analyzing their similarities and differences. Methods: Three dimensional reconstruction of the arteries in breast were performed in 50 patients between September 2015 and August 2017 with breast hypertrophy by computed tomographic angiography (CT angiography). The distribution pattern and the source direction of each main blood vessel was observed, counted and analyzed. Then, the data of breast hypertrophy patients were compared with the previous data about nipple-areola blood supply in normal population (the definition of main vessel: entering the breast gland or reaching the nipple-areola surrounding area, and diameter larger than 1 mm). Statistical description was taken for comparison. Results: 135 main vessels were observed in 100 breasts (50 patients). They mainly originate from the internal thoracic artery (69, 51.1%), lateral thoracic artery (37, 27.4%) and thoracoacromial artery(16, 11.9%), as well as a small amount from the brachial artery (7, 5.2%) and axillary artery(6, 4.4%). No main supply vessels from the posterior intercostal artery have been found. The patterns of breast blood supply varied among individuals, and high asymmetry ratio in the same individual was also observed. The internal superior (left: 30.7%, right: 34.2%) and superior lateral quadrant (Left: 29.2%, Right: 20%) of the breast was the most likely area for the main vessel to pass, followed by the breast lateral (Left: 16.9%, Right: 18.5%), lower inner (Left: 4.6%, Right: 5.7%), central (Left: 4.6%, Right: 4.2%), and superior (Left: 1.5%, Right: 2.8%). Differences existed in main vessels between normal breasts and hypertrophic breasts, either for source arteries or the distribution of breast. There was no main blood supply from the intercostal arteries or across the outer inferior quadrant. Conclusions The blood supply of the nipple-areola is not completely consistent between the hypertrophic breast and the normal size breast, and the blood supply pattern of the hypertrophic breasts is complex and diverse. CT angiography might be used before breast reduction surgery for clarifying the direction of the main vessels, so as to preserve more blood supply for nipple-areola, and to prevent nipple-areola necrosis.