Biomedical Research ; China ; Freeze Drying ; Hepatitis A Vaccines ; adverse effects ; immunology ; Humans ; Safety ; Vaccines, Attenuated ; adverse effects ; immunology

The peak level of growth hormone (GH) stimulated by insulin tolerance test (ITT) is thegold standard for diagnosis of growth hormone deficiency in adults.This study was aimed to explore the factors influencing GH response to ITT in 50 healthy adults.The results showed that the nadir or decreased amplitude of blood glucose was not related to GH peak level.In multivariable analysis,the GH level stimulated by ITT was negatively associated with body mass index(P＜0.01),but there was no any association with age,gender,and waist circumference.

Objective To investigate the association between extramural vascular invasion (EMVI) detected by multi-detectors computed tomography (MDCT) with contrast enhanced (ceMDCT) and clinicopathologic characteristics in patients with colon cancer.Methods Between February 2009 and December 2013,patients with histologically proven primary colon cancer and undergoing curative resection were included in this retrospective study.According to American Joint Committee on Cancer TNM staging system,patients of stage Ⅱ and Ⅲ were included in this study.EMVI status detected by MDCT (ctEMVI) was defined according to the EMVI scores.Chi-square test was used to analyze the association between clinicopathologic characteristics and ctEMVI.Results 165 stage Ⅱ and stage Ⅲ patients were included in this study as confirmed by pathology based on AJCC.Positive ctEMVI was demonstrated in 51 patients (34.5％,51/165).There were significant association between positive ctEMVI and age ＜ 65 years (x2 =4.810,P =0.031),ceMDCT defined tumor stage (x2 =17.911,P =0.000),ceMDCT defined metastatic lymph node (x2 =5.436,P =0.022),tumor size≥5 cm (x2 =3.799,P =0.036) and pathological T stage (x2 =13.346,P =0.001).Conclusions EMVI,detected by ceMDCT,is significantly associated with age,tumor size and T staging in colon cancer.

BACKGROUNDThe dust mites, which are mostly represented by Dermatophagoides spp. (Acari: Pyroglyphidae), are the major sources of indoor allergens. Identification and characterization of these mite allergen molecules are an important step in the development of new effective diagnostic procedures and possible therapeutic strategies for allergic disorders associated with dust mites.

METHODSTotal RNA was extracted from Dermatophagoides farinae. The gene coding for Der f 3 was amplified by RT-PCR with the primers designed based on previous sequence published in GenBank. The target gene was cloned intermediately into pMD19-T plasmid and finally into plasmid pET28a (+), expressed in E. coli BL21 at the aid of the inducer isopropyl-D-thiogalactopyranoside (IPTG). The physicochemical properties, spatial structure of the allergen were analyzed with bioinformatics software.

RESULTSThe cDNA coding for group 3 allergen of Dermatophagoides farinae from China was cloned and expressed successfully. Sequencing analysis showed that there were nineteen mismatched nucleotides in five Der f 3 cDNA clones in comparison with the reference (GenBank Accession No. AY283291), which resulted in deduced amino acid sequence incompatibility in eleven residues. Bioinformatics analysis revealed that the Der f 3 pro-protein was an extracellular hydrophobic protein, consisting of 259 amino acids with a 16 amino acid signal peptide. The protein was deduced to have three chymotrypsin active sites (53-68 AA, 108-122 AA and 205-217 AA), one N-glycosylation site, one cAMP- and cGMP-dependent protein kinase phosphorylation site, four protein kinase C phosphorylation sites, two casein kinase II phosphorylation sites, and five N-myristoylation sites.

CONCLUSIONSDer f 3 is an extracellular hydrophobic protein which possesses multiple activation and phosphorylation sites. Polymorphism may exist in the Der f 3 gene but this needs to be further confirmed in the future.

Allergens ; chemistry ; genetics ; metabolism ; Amino Acid Sequence ; Animals ; Antigens, Dermatophagoides ; chemistry ; genetics ; metabolism ; Blotting, Western ; Cloning, Molecular ; Computational Biology ; Dermatophagoides farinae ; genetics ; Escherichia coli ; genetics ; metabolism ; Molecular Sequence Data ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Homology, Amino Acid

Background The dust mites,which are mostly represented by Dermatophagoides spp.(Acari:Pyroglyphidae),are the major sources of indoor allergens.Identification and characterization of these mite allergen molecules are an important step in the development of new effective diagnostic procedures and possible therapeutic strategies for allergic disorders associated with dust mites.Methods Total RNA was extracted from Derrnatophagoides farinae.The gene coding for Der f 3 was amplified by RT-PCR with the primers designed based on previous sequence published in GenBank.The target gene was cloned intermediately into pMD19-T plasmid and finally into plasmid pET28a(+),expressed in E.coli BL21 at the aid of the inducer isopropyI-D-thiogalactopyranoside(IPTG).The physicochemical properties,spatial structure of the allergen were analyzed with bioinformatics software.Results The cDNA coding for group 3 allergen of Dermatophagoides farinae from China was cloned and expressed successfully.Sequencing analysis showed that there were nineteen mismatched nucleotides in five Der f 3 cDNA clones in comparison with the reference(GenBank Accession No.AY283291),which resulted in deduced amino acid sequence incompatibility in eleven residues.Bioinformatics analysis revealed that the Der f 3 pro-protein was an extracellular hydrophobic protein,consisting of 259 amino acids with a 16 amino acid signal peptide.The protein was deduced to have three chymotrypsin active sites(53-68 AA,108-122 AA and 205-217 hA),one N-glycosylation site,one cAMP-and cGMP-dependent protein kinase phosphorylation site,four protein kinase C phosphorylation sites,two casein kinase Ⅱ phosphorylation sites,and five N-myristoylation sites.Conclusions Der f 3 is an extracellular hydrophobic protein which possesses multiple activation and phosphorylation sites.Polymorphism may exist in the Der f3 gene but this needs to be further confirmed in the future.

BACKGROUND:Vertebral compression fracture is a common disease in the current orthopedic field.However,the occurrence of re-fracture in neighboring vertebrae after surgery is a problem that cannot be ignored,which has a serious impact on the normal life of patients. OBJECTIVE:The aim of this study is to establish four postoperative models with different recovery heights using computed tomography images.By using finite element analysis,we derived the stresses on the neighboring vertebrae at different recovery heights and further explored the importance of postoperative recovery of the height of the injured vertebrae. METHODS:A finite element model of the thoracolumbar spine(T11-L3)was established and validated,on the basis of which four postoperative finite element models of L1 with different recovery heights of 100%,80%,60%,and 40%were constructed,in which the cement capacity varied with the recovery height.The specific models are as follows:Model 1 was the postoperative model with normal recovery height,and the cement capacity was 8.3 mL.Model 2 was the postoperative model in which 20%of the anterior height of the L1 was removed and the posterior convexity angle became 10.41°,and the cement capacity was 6.9 mL.Model 3 was the postoperative model in which 40%of the anterior height of the L1 was removed and the posterior convexity angle became 20.17°,and the cement capacity was 4.7 mL.Model 4 was a postoperative model with 60%of the L1 anterior height removed and the posterior convexity angle changed to 28.85°,with a cement capacity of 3.6 mL.For evaluation of the postoperative model,we applied a moment of 7 Nm and an axial force of 500 N.The followings were recorded and analyzed:peak stresses in the L2 upper endplate and T12 lower endplate;peak stresses in the L2 and T12 cancellous bone. RESULTS AND CONCLUSION:(1)The highest peak stresses for each condition of the L2 upper endplate,T12 lower endplate,L2 cancellous bone,and T12 cancellous bone occurred in Model 1 and Model 4.In particular,the T12 lower endplate,except for the posterior extension condition,the anterior flexion,left and right lateral bending,and left and right rotation conditions all reached their highest peak stresses in Model 4,with stresses of 50.3,33.1,44.9,34.3,and 31.9 MPa.(2)Based on the peak stresses in the adjacent vertebral endplates and cancellous bone,after excluding Model 1 and Model 4,the minimum peak stresses for most of the conditions appeared in the Model 2,and the minimum peak stresses appeared in the Model 2 in 66.6%of the cases,especially in the upper endplates of the L2 and cancellous bone except for the posterior extension condition,the minimum peak stresses all appeared on the Model 2.(3)Therefore,controlling the recovery height at about 100%and 40%of the original height was a dangerous recovery height,which had a greater impact on the neighboring vertebrae.Controlling the recovery height at about 80%of the original height may be a more ideal choice.With a recovery height of about 80%of the original height,the adjacent vertebrae are subjected to less stress,thus reducing the risk of re-fracture of the adjacent vertebrae in the patient.

Objective: To investigate the clinicopathologic and molecular features of the rare cribriform morular variant of papillary thyroid carcinoma (CMV-PTC). Methods: The clinicopathologic data of 10 patients with CMV-PTC were retrospectively reviewed. Immunohistochemical (IHC) staining was done using LSAB method. DNA sequencing for APC were applied using Sanger method. BRAF V600E mutation was examined using ARMS method. The cytological, morphological, IHC and molecular features were analyzed. Results: All patients were female at an average age of 27 years old. The tumors were mostly located in the right lobe of thyroid. Fine needle aspiration cytology was performed in three patients; two were diagnosed as suspicious for PTC and one as PTC. Nine tumors presented as solitary nodule and two as multiple nodules in both lobes. Infiltration was demonstrated in three cases. The average size was 2.6 cm. The neoplastic cells were arranged in papillary, cribriform, solid and glandular patterns, with rare or without colloid inside the lumen. The number of morula varied, ranging from zero to many. The neoplastic cells were variably enlarged, showing round, oval or spindle shape. Nuclear irregularity was identified as irregular membrane, nuclear grooves or pseudoinclusion, but no typical ground glass feature. Peculiar nuclear clearing could be observed in the morular cells. IHC staining showed the neoplastic cells were negative for thyroglobulin and p63, but positive for TTF1, cytokeratin 19 and estrogen receptor. Diffuse staining with cytokeratin was seen in the neoplastic cells and the morula. Specific cytoplasmic and nuclear staining of β-catenin was seen in the neoplastic cells but not the morula. Ki-67 proliferation index was 1%-30%. No recurrence or metastasis was observed. One patient was demonstrated to harbor both somatic and germline mutations of the APC gene, who was found to have adenomatous polyposis and her mother died of colonic carcinoma. No BRAF V600E mutation was detected. Conclusions CMV-PTC is rare and shows atypical cytological and clinicopathological features, and it is easily misdiagnosed.TG, TTF1, ER and β-catenin are specific IHC markers for CMV-PTC. The morula is negative for cytokeratin 19, in contrast to squamous metaplasia. Although CMV-PTC has indolent clinical behavior, a definite diagnosis is necessary to rule out the possibility of APC gene mutation and related extra-thyroidal neoplasm, such as FAP and Gardner syndrome.

Objective:To observe the clinical characteristics of esophageal reflux after total gastrectomy (ERATG), and to explore the mechanism of occurrence.Methods:Fourteen gastric cancer patients who underwent total gastrectomy were prospectively enrolled in this study. The postoperative symptoms were observed and recorded and 24 h MII-pH with pH monitoring was performed to investigate the characteristics of postoperative reflux.Results:After total gastrectomy patients were with different degrees of ERATG as heartburn, appetite loss, chest tightness and belching. The overall nature of ERATG is mainly weak acid, with a pH between 4 and 7. ERATG involved esophageal-jejunal anastomosis and a length of esophagus 7 cm above the anastomosis. Patients with typical reflux symptoms had a lower pH minimum in the upright position than those without typical symptoms[(4.76±0.71) vs.(5.68±0.37), t=2.866, P<0.05]. Patients with typical reflux symptoms had a higher frequency of reflux of mixed liquid and liquid-air reflux than those without typical symptoms[liquid(31.25±29.76) vs.(4.50±9.14), t=0.011, P<0.05; liquid-air(19.50±12.99) vs.(2.00±2.61), t=0.004, P<0.05]. Conclusion:ERATG is mainly a upward reflux of weakly acidic gas, with typical symptoms of heartburn, appetite loss, chest tightness and belching. Patients with typical symptoms usually have lower pH in the upright position.

Objective:To explore the effect of tumor deposit (TD) on the prognosis of patients with stage Ⅲ colon cancer after radical resection.Methods:The clinicopathological data of patients with stage Ⅲ colon cancer after radical surgery at the Department of Gastrointestinal Surgery, Peking University People's Hospital from Jan to Dec 2015 were analyzed collected. Clinicopathological characteristics such as tumor location, degree of differentiation, mismatch repair status, lymphatic and venous invasion, and preoperative CEA and CA19-9 levels were used to study the effect of TD on the postoperative survival of patients.Results:Among the 155 patients with stage Ⅲ colon cancer, 37 (23.9%) had tumor deposits. The incidence of tumor deposits was higher in patients with intravascular tumor thrombus and preoperative serum CA19-9 elevation ( χ2=9.567, P=0.002; χ2=11.561, P=0.003); Patients with tumor deposits had worse overall survival and disease-free survival than those without cancer nodules (OS: P=0.029, DFS: P=0.025). Multivariate COX analysis found that tumor deposit was an independent risk factor for postoperative overall survival and disease-free survival ( HR=1.990, 95% CI: 1.032-3.835, P=0.040; HR=2.416, 95% CI : 1.205-3.820, P=0.009). Conclusions:Tumor deposit is an independent risk factor affecting postoperative overall survival and disease-free survival in patients with stage Ⅲ colon cancer. For patients with lymph node metastasis, incorporating TD into TNM staging can more accurately predict the postoperative prognosis.

BACKGROUND:The elastic modulus of traditional bone implants is large and does not match the elastic modulus of human bone,which will cause a stress shielding effect and lead to bone resorption.The trabecular scaffold of the triply periodic minimal surface with radial gradient has elastic modulus matching with human cancellous bone,and its yield strength is greater than that of human cortical bone,which provides a new choice for the design of bone scaffold. OBJECTIVE:Triply periodic minimal surface structure with radial gradient was constructed by the implicit surface method.The sample was manufactured by laser selective melting technology,and the quasi-static compression test was carried out to obtain trabecular scaffolds with mechanical properties matching human bones. METHODS:Four types of the trabecular scaffolds of the triply periodic minimal surface with a radial gradient of G,I,P and D were established by the implicit surface method.Samples were manufactured by laser selective melting technology.We observed the surface morphology of the molded sample,evaluated the molding quality,conducted a quasi-static compression test,and evaluated the mechanical properties of the samples. RESULTS AND CONCLUSION:The quasi-static compression test results showed that compared with the four triply periodic minimal surface scaffolds,the platform stress of the G scaffold had less fluctuation and no failure or fracture,indicating that the G scaffold had the best plasticity.The mechanical properties of the G scaffolds with 45%,55%and 65%porosities were analyzed.It was found that the elastic modulus of G scaffolds with 55%porosity was within the range of elastic modulus of human cancellous bone(0.022-3.7 GPa),and the yield strength was close to the maximum yield strength of human cortical bone(187.7-222.3 MPa).In conclusion,G triply periodic minimal surface scaffold with 55%porosity can reduce the stress shielding effect,bear a higher body load,improve the stability of the implant,and prolong the service life of the implant.