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【Objective】 To explore the application of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) in the genotyping of difficult blood typing samples, and to provide evidence for clinical blood transfusion. 【Methods】 Three ambiguous blood group samples, submitted to Shanghai Blood Center by Shanghai regional hospitals, were studied, of which Sample1 included the proband and his parents. Serological methods were used to perform blood group typing, direct antibody test, unexpected antibody screening and identification test. Blood group genotyping was performed by using the MALDI-TOF MS detection systeme stablished in our laboratory. Sanger sequencing was used to confirm gene mutation sites, and serological or flow methods were used to verify specific samples′ phenotype. 【Results】 Serological results indicated the existence of antibodies against high frequency antigens in sample 1 (including proband and her mother), 2 and 3. The genotyping results of MALDI-TOF MS showed that the proband of sample 1 was Di(a+ b+ ), her father was Di(a-b+ ), her mother was Di(a+ b-), sample 2 was p, and sample 3 was Jr(a-). Sequencing results of three samples were consistent with mass spectrometry typing results. Serological results showed that sample 2 had a p phenotype. The flow cytometry results suggested that sample 3 had a Jr(a-) phenotype. 【Conclusion】 For the first time, we applied MALDI-TOF MS technology to blood type genotyping of ambiguous clinical samples in China. Compared with other genotyping methods such as PCR-SSP, MALDI-TOF MS has the advantages of rapid detection, high throughput and high specificity, which would contribute to identification of difficult blood typing samples in the future, as well as rare blood group screening.
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Objective:To investigate the efficacy and safety of cortical-sparing adrenalectomy (CSA) in the treatment of bilateral pheochromocytoma.Methods:The clinical data of 20 patients with bilateral pheochromocytoma treated in Xiangya Hospital of Central South University from January 2004 to December 2019 were analyzed retrospectively, including 10 males and 10 females. The average age of onset was 32.5 (8-51) years. 3 cases had a family history of pheochromocytoma. There were 14 and 6 patients with bilateral synchronous and metachronous onset, respectively. The mean value of vanilmandelic acid (VMA) in 20 cases was (106.4 ± 60.0) μ mol/24h. Preoperative enhanced CT showed a soft tissue mass with uneven enhancement in the adrenal region, with low-density necrosis, which suggested the diagnosis of Pheochromocytoma. All 20 cases underwent CSA under general anesthesia. In 14 cases of bilateral synchronous disease, 9 cases underwent simultaneous operation and 5 cases underwent staged operation; 6 patients with metachronous disease underwent bilateral tumor resection successively. Laparoscopic surgery was performed in 18 cases and open surgery in 2 cases. Through the abdominal or retroperitoneal approach, open the fat capsule around the upper pole of the kidney, free the medial edge of the upper pole of the kidney, expose the adrenal gland and tumor, completely remove the tumor and capsule, ensure that the adrenal tissue is 3-5 mm away from the cutting edge of the tumor, and the reserved cortical size is at least 1 / 3 of the ipsilateral adrenal gland. The central adrenal vein was preserved as much as possible to reduce the damage to the adrenal vascular bed. The operation related data, intraoperative monitoring records, postoperative complications and long-term follow-up results were recorded.Results:All the 20 cases were successfully completed without tumor rupture. The operation time of simultaneous operation and staged operation were (242.3 ± 61.0) min and (137.9 ± 60.3) min, respectively. The number of patients admitted to ICU after operation was 7 and 2, respectively ( P<0.05); The intraoperative bleeding volume was (528.6 ± 355.7) ml and (277.8 ± 264.7) ml, the number of blood transfusion cases were 5 and 2 cases, and the average hospital stay was (7.4 ± 2.0) d and (7.8 ± 3.3) d, respectively ( P>0.05). 20 cases took glucocorticoid orally (prednisone 5 mg, once every 12 hours) after operation. There was no obvious manifestation of adrenocortical dysfunction and Addison's crisis. The hormone was stopped gradually from 2 weeks to 1 month after operation. The average follow-up was 5.4 (1.0-16.0) years. There were 3 cases of recurrence and no metastasis. Gene detection was performed in 10 cases after operation, and 7 cases carried pheochromocytoma RET and VHL pathogenic gene mutations (RET in 2 cases and VHL in 5 cases). Conclusion:Although CSA has a certain risk of recurrence, it avoids hormone replacement and does not increase the risk of metastasis and death. It is recommended for the treatment of hereditary pheochromocytoma, especially bilateral pheochromocytoma.
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Objective:To study the effects of FcγRIIb gene modified dendritic cells on liver function and rejection after orthotopic liver transplantation in rats.Methods:The recombinant lentivirus expression vector TRE-FcγRIIb containing Lewis rat FcγRIIb gene was constructed by gene cloning technique. TRE-FcγRIIb expression virus and TET-on regulatory virus were packaged and co-infected with immature dendritic cells derived from DA rat bone marrow. The expression of FcγRIIb was detected. Donor rats DA and recipient Lewis were paired according to body weight and then randomly divided into three groups. The control group (group A) did not receive any pretreatment. The Lewis rats in group B were treated with cyclosporin A on the 2nd day after liver TX. Immature dendritic cells derived from bone marrow of DA rats were injected intravenously one day before liver TX in FcγRIIb gene modified immature dendritic cells (1×10 6 cells)group (group C). Blood and liver tissue were biopsied 7 days after operation to detect liver function and pathological changes. Results:The abnormal liver function in FcγRIIb gene modified immature dendritic cells group (group C) was significantly lower than that of control group 7 days after operation ( P<0.05), and the liver pathological rejection of FcγRIIb gene modified immature dendritic cells group (group C) was significantly lower than that of control group 7 days after operation ( P<0.05). The average survival days of rats in the control group was 12.8 days; the average survival days of rats in the cyclosporin A group was 65.3 days; the average survival days of rats in the FcγRIIb gene-modified immature dendritic cell group was 58.5 days. Conclusion:FcγRIIb gene modified immature dendritic cells can ameliorate liver dysfunction and acute liver rejection after orthotopic liver transplantation.
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Objective To study the characteristics,operative timing,repair methods for iatrogenic bile duct injury caused by laparoscopic cholecystectomy.Methods Clinical data of 30 cases of iatrogenic bile duct injury found during or after laparoscopic cholecystectomy were studied retrospectively.Results There were type Ⅱ 1 bile duct injury in 9 patients,type Ⅱ 2 in 4 patients,type Ⅱ 3 in 2 patients,and type Ⅱ 4 in 15 patient.17 cases underwent immediate repair,13 cases did early repair.All cases followed up from 5 months to 10 years,the follow-up rate was 83.3％.1 case with end-to-end anastomosis and biliary stent underwent Roux-en-Y choledochojejunostomy for biliary stenosis 13 months postoperatively,4 cases underwent secondary Roux-en-Y choledochojejunostomy because of obstructive jaundice and the cicatricial anastomotic orifice stenosis one to one and half years after primary repairs.The remaining cases were doing well up to the end of the follow-up.Conclusion Most iatrogenic bile duct injury after laparoscopic cholecystectomy were high in position,while Roux-en-Y choledochojejunostomy is the mainstay of repair.
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Objective To investigate the significance of calcification in thyroid node for diagnosis of thyroid carcinoma.Method Retrospective analysis of 107 thyroid nodules' pre-operative ultrasonic and postoperative pathologic results.Results Total ultrasonic thyroid calcification ratio was 27.1%(29/107),which in benign samples was lower than Ihat in malignant samples(17.2%vs 70.0%,P＜0.01).Micro-calci-fication ratio in benign samples Was lower than thai:in malignanl samples(8.0%vs 50.0%,P＜0.01).Conclusion The ralio of thyroid carcinoma with calcification is higher,so the detection of thyroid carcinoma,especially micro-single-calcification should be significant.