Objective:To prepare and test tetrameric sH-2K~d-HBc complex for the further measurement of the specific CTL response.Methods:PE labled streptavidin with 4 biotinylated binding sites can bind to 4 biotinylated monomer to form the corresponding tetramer.Mice were immunized via different methods of genetic immunization by use of the construted pcDNA3-C plasmid to get the specific CTLs.Then our prepared tetramer was applied to stain the specific CTLs by the analysis of flow cytometry.Results:We applied our prepared tetramer to stain the cells from the experimental groups and control group.The results showed the tetramer was able to discriminate the frequencies of specific CTL induced by the three immunol methods(0.24%,0.26%,0.36% vs 0.07%,P≤0.05).This demonstrated that the prepared tetramer could bind its targets specifically and efficiently.The three immunol methods induced different levels of immune responses.Compared with the traditional muscle injection,gene gun induced weaker humoral immune response and stronger cellular immune response,and hydrodynamic injection induced the strongest humoral and cellular immune responses.Conclusion:Have successfully constructed the sH-2K~d-HBc tetramer.The techniques and methods can be used for preparation of tetramers of other types of MHCⅠ molecules.

Objective To explore the computed tomography (CT)manifestations of adrenocortical oncocytoma (ACO)for better understanding the disease and improving its diagnosis accuracy.Methods The CT manifestations of 9 cases with adrenocortical onco-cytomas confirmed by surgery were retrospectively reviewed and compared with pathological results.Results Five of cases were lo-cated in left side while 4 cases were in right side.Well defined and round or oval border could be found in all the cases.Of all the ca-ses,3 cases were solid lesions with equal density,< 3 cm in diameter,and mild consistent intensification after enhancement.Cystic lesion was detectd in 6 cases with > 3 cm in diameter.The mixed cystic and solid components with polycystic changes could be found on CT image,and remarkable intensification on solid lesion while no intensification on cystic lesion after the enhancement. Moreover,patchy or cotton-like shape could be found on solid lesion in 4 cases and 1 case with island like intensification;reticular pattern could be found in the central of lesion in 3 cases and 1 case with stellate shape.Pathological observation showed that the on-cotytomas were comprised of cells with abundant eosinophilic cytoplasm and necrotic and fibrous capsule without pathological mitosis could be found in some of lesion.Conclusion The characteristic CT features of adrenocortical oncotytoma has a worse specificity,its diagnosis was dependent on pathologic examination.

Objective To analyze the expression levels and clinical significance of N-myc down-stream-regulated gene 3(NDRG3)and semaphoring 3A(SEMA3A)in elderly patients with acute ischemic stroke(AIS).Methods A total of 100 elderly AIS patients admitted to Department of Geriatrics of Liaocheng People's Hospital from September 2020 to September 2022 were included as the study group.According to their NIHSS score at admission,they were divided into mild(34 cases),moderate(31 cases)and severe(35 cases)subgroups.All patients were followed up for 3 months after discharge.And they were assigned into good prognosis group(69 cases)and poor prognosis group(31 cases)based on the modified Rankin scale score.Another 100 healthy individ-uals who underwent physical examination in our hospital during the same period were recruited as the control group.Western blotting was used to detect the protein expression of NDRG3 and SEMA3A in peripheral blood mononuclear cells(PBMC).ELISA was applied to measure the con-tents of VEGF,TGF-1,TNF-α,and IL-17 in peripheral blood samples.Spearman rank correlation analysis was performed to analyze the correlation of NDRG3 and SEMA3A levels with NIHSS score,and ROC curve was plotted to analyze the values of NDRG3 and SEMA3A in predicting poor prognosis in elderly AIS patients.Results The expression levels of NDRG3 and SEMA3A in PBMC were obviously higher in the study group than the control group(1.11±0.16 vs 0.76± 0.13,0.78±0.13 vs 0.42±0.09,P＜0.01).The levels in the mild,moderate and severe subgroups were significantly higher than that of the control group(P＜0.01).The poor prognosis group had statistically higher expression levels of NDRG3 and SEMA3A than the good prognosis group(P＜0.01).Spearman rank correlation analysis showed that the NIHSS score was positively corre-lated with the expression levels of NDRG3 and SEMA3A in elderly AIS patients(r=0.597,P＜0.01;r=0.618,P＜0.01),while the NDRG3 level was positively correlated with that of SEMA3 A(r=0.477,P＜0.01).ROC curve analysis indicated that the AUC value of combined NDRG3 and SEMA3A levels was superior to that of NDRG3 and SEMA3A alone in predicting of poor progno-sis(0.962 vs 0.861,0.880,P＜0.01).Conclusion The levels of NDRG3 and SEMA3A proteins are up-regulated in elderly AIS patients,and are closely associated with the severity and prognosis of the disease.

OBJECTIVETo identify the casual mutation of a Chinese pedigree with hypertrophic cardiomyopathy (HCM), and to analyze the genotype-phenotype relationship.

METHODSThe coding exons of 26 reported disease genes were sequenced by targeted resequencing in the proband and the identified mutation were detected with bi-directional Sanger sequencing in all family members and 307 healthy controls. The genotype-phenotype correlation was analyzed in the family.

RESULTSA missense mutation (c.2191C > T, p. Pro731Ser) in the 20th exon of MYH7 gene was identified. This mutation was absent in 307 healthy controls and predicted to be pathogenic by PolyPhen-HCM. Totally 13 family members carried this mutation, including 10 patients with HCM and 3 asymptomatic mutation carriers. The proband manifested severe congestive heart failure and 8 patients expressed various clinical manifestations of heart failure, including dyspnea, palpitations, chest pain, amaurosis or syncope. Five patients were diagnosed as HCM at the age of 16 or younger. One family member suffered sudden cardiac death.

CONCLUSIONSThe Pro731Ser of MYH7 gene mutation is a causal and malignant mutation linked with familiar HCM.

Adolescent ; Asian Continental Ancestry Group ; Base Sequence ; Cardiomyopathy, Hypertrophic ; ethnology ; genetics ; Death, Sudden, Cardiac ; Exons ; Humans ; Mutation, Missense ; Myosin Heavy Chains ; genetics ; Pedigree ; Phenotype ; Research Design ; Ventricular Myosins

Objective To verify the leaf positioning accuracy of multi-leaf collimator (MLC) in intensity modulated radiation therapy (IMRT) by using radiochromic films to provide references for IMRT quality control.Methods Medical linear accelerators of different designs,owned by 7 first-class hospitals at grade-3 in Hubei,were selected for this verification study.Five strip picket fence pattern was created by treatment planning system (TPS).Each strip was 0.6 cm wide with a 3 cm strip separation.MLC leaf positions,positioning bias and opening widths were then measured with EBT2 radiochromic films.Results According to IAEA standards,the film-measured and TPS planned MLC leaf position difference was ±0.5 mm.The difference of MCL strip position of No.5 and No.7 accelerator was 0.7 and-1.0 mm,respectively,not in line with the IAEA standards.The difference of film-measured MLC leaf position between each pairs and all pairs of leaves of 7 accelerators were all within ± 0.5 mm,in line with the IAEA requirements.As required by IAEA,the difference of opening width of MLC leave of each pair relative to the averaged widths of all pairs should be within ±0.75 mm.The filem-measured values from 7 accelerators ranged from-0.6 to 0.5 mm,all in line with the IAEA standards.Standard deviation of opening width of all leaves were required to be within 0.3 mm.The measured values from 7 accelerators ranged from 0.1 to 0.2 mm,in line with the IAEA standard.Conclusions The use of EBT2 radiochromic films to verify MLC leaf positioning accuracy is an important means of quality control owing to its simplicity and high measurement accuracy.It is recommended for verification use at a large scale.

Humans ; Coronary Artery Disease/surgery* ; Percutaneous Coronary Intervention ; Heart ; Treatment Outcome ; Coronary Angiography

Objective To identify the casual mutation of a Chinese pedigree with hypertrophic cardiomyopathy (HCM),and to analyze the genotype-phenotype relationship.Methods The coding exons of 26 reported disease genes were sequenced by targeted resequencing in the proband and the identified mutation were detected with bi-directional Sanger sequencing in all family members and 307 healthy controls.The genotype-phenotype correlation was analyzed in the family.Results A missense mutation (c.2191C>T, p.Pro731Ser) in the 20th exon of MYH7 gene was identified.This mutation was absent in 307 healthy controls and predicted to be pathogenic by PolyPhen-HCM.Totally 13 family members carried this mutation , including 10 patients with HCM and 3 asymptomatic mutation carriers.The proband manifested severe congestive heart failure and 8 patients expressed various clinical manifestations of heart failure , including dyspnea , palpitations , chest pain , amaurosis or syncope.Five patients were diagnosed as HCM at the age of 16 or younger.One family member suffered sudden cardiac death.Conclusions The Pro731Ser of MYH7 gene mutation is a causal and malignant mutation linked with familiar HCM.

Background::Risk assessment and treatment stratification for three-vessel coronary disease (TVD) remain challenging. This study aimed to investigate the prognostic value of left atrial volume index (LAVI) with the Synergy Between Percutaneous Coronary Intervention with Taxus and Cardiac Surgery (SYNTAX) score II, and its association with the long-term prognosis after three strategies (percutaneous coronary intervention [PCI], coronary artery bypass grafting [CABG], and medical therapy [MT]) in patients with TVD.Methods::This study was a post hoc analysis of a large, prospective cohort of patients with TVD in China, that aimed to determine the long-term outcomes after PCI, CABG, or optimal MT alone. A total of 8943 patients with TVD were consecutively enrolled between 2004 and 2011 at Fuwai Hospital. A total of 7818 patients with available baseline LAVI data were included in the study. Baseline, procedural, and follow-up data were collected. The primary endpoint was major adverse cardiac and cerebrovascular events (MACCE), which was a composite of all-cause death, myocardial infarction (MI), and stroke. Secondary endpoints included all-cause death, cardiac death, MI, revascularization, and stroke. Long-term outcomes were evaluated among LAVI quartile groups. Results::During a median follow-up of 6.6 years, a higher LAVI was strongly associated with increased risk of MACCE (Q3: hazard ratio [HR] 1.20, 95% confidence interval [CI] 1.06-1.37, P = 0.005; Q4: HR 1.85, 95%CI 1.64-2.09, P <0.001), all-cause death (Q3: HR 1.41, 95% CI 1.17-1.69, P <0.001; Q4: HR 2.54, 95%CI 2.16-3.00, P <0.001), and cardiac death (Q3: HR 1.81, 95% CI 1.39-2.37, P <0.001; Q4: HR 3.47, 95%CI 2.71-4.43, P <0.001). Moreover, LAVI significantly improved discrimination and reclassification of the SYNTAX score II. Notably, there was a significant interaction between LAVI quartiles and treatment strategies for MACCE. CABG was associated with lower risk of MACCE than MT alone, regardless of LAVI quartiles. Among patients in the fourth quartile, PCI was associated with significantly increased risk of cardiac death compared with CABG (HR: 5.25, 95% CI: 1.97-14.03, P = 0.001). Conclusions::LAVI is a potential index for risk stratification and therapeutic decision-making in patients with three-vessel coronary disease. CABG is associated with improved long-term outcomes compared with MT alone, regardless of LAVI quartiles. When LAVI is severely elevated, PCI is associated with higher risk of cardiac death than CABG.