BACKGROUND:An effective freezing-thawing technique is crucial for the clinical application of human umbilical cord mesenchymal stem cells(UC-MSCs).OBJECTIVE:To investigate biological characteristics of UC-MSCs after cryopreservation.METHODS:UC-MSCs were isolated from human umbilical cord and frozen in liquid nitrogen.The survival rate and the suppressive effect of γ-interferon(IFN-γ)of cryopreserved-thawed and fresh human UC-MSCs were compared.Furthermore,the multiple potentials and phenotype of UC-MSCs were estimated after cryopreservation.RESULTS AND CONCLUSION:There was no significant difference between cryopreserved-thawed and fresh human UC-MSCs on the survival rate and the suppressive effect of IFN-γ of peripheral blood mononuclear cells(PBMCs).After cryopreservation,human UC-MSCs had the potential differentiation and the phenotype of mesenchymal stem cells.

Objective To identify lesion patterns and stroke mechanisms in middle cerebral artery (MCA) territory using early diffusion-weighted imaging (DWI) combined with CTA as well as EKG and echocardiography.Methods One hundred and forty-eight acute ischemic stroke patients who had (1) symptomatic lesions located in the unilateral MCA territory on DWI performed within 1 week of symptom onset,and (2) either corresponding MCA disease,internal carotid artery (ICA) disease,MCA & ICA disease or cardio embolism (CE),or (3) neither corresponding MCA disease,ICA disease,nor CE which were taken as group of negative results (NR),were reviewed.Acute DWI lesion patterns were classified as (1) single (small perforator < 2 cm;large perforator ≥2 cm;pial;large territorial;border-zone) and (2) multiple according to principle of single-blind.Results There were 12 types of lesions in MCA territory.Distribution of lesion patterns in different stroke subtypes might be different (χ2= 55.88,P = 0.004).No specific pattern could be found in patients with MCA disease,ICA disease,MCA & ICA disease or CE.Big perforator infarcts might be more common in patients with MCA disease than with ICA disease and CE.Compared with negative group,concomitant perforator and pial infarcts were more common in patients with ICA disease (7/27,χ2=6.61,P <0.05),especially with severe stenosis or occlusion (5/16,χ2=7.32,P < 0.05);No specific pattern could be found in patients with MCA disease or CE.Concomitant perforator,pial,with border-zone infarcts (6/30,χ2= 6.41,P <0.05),and concomitant perforator with border-zone infarcts (4/30,χ2= 5.59,P < 0.05) were more often in patients with severe stenosis or occlusion of MCA.Conclusion Different lesion patterns may indicate different mechanisms of stroke such as hypoperfusion and arterial embolism could be coexistent in MCA territory.The relationship has not been identified perfectly.

Objective To evaluate the accuracy of presurgically resting-state fMRI(rs-fMRI) with subject order-independent group independent component analysis(ICA), compared to electric cortical stimulations. Methods Twenty-three patients with the lesion in motor area, which were recorded by our hospital from Jan, 2014 to Dec, 2015, were collected as the study sample. The data of 9 patients were excluded because of excessive head motion. As a result, 14 patients were included in this study. Rs-fMRI data before the surgery and the results of electric cortical stimulations were collected. Results All of this 14 patients were preoperatively located by rs-fMRI with SOI-GICA, including all the SMA and the ipsilesional primary motor area. On the side with lesion, the number of functional location of motor area was decreased compared with healthy side. Evaluate the accuracy of ICA by comparing the coincidence rate of these two techniques, based on the standardized electrical cortical stimulation in operation. The completely concordance between rs-fMRI with the SOI-GICA and electrical cortical stimulation in operation was 11 (11/14). Meanwhile the basically concordance of corresponded case was 3(3/14). Conclusions Rs-fMRI with the ICA has a relatively high accuracy rate in localizing motor area. Rs-fMRI has a remarkably referential contribution to the presurgically function assessment and surgical planning in implementation.

Objective To investigate the enhancement features of hepatic tumors between dynamic three-dimensional contrast-enhanced ultrasonography (3D-CEUS) and two-dimensional contrast-enhanced ultrasonography (2D-CEUS).MethodsTotally,65 patients with 67 focal liver lesions were examined with 2D-CEUS and dynamic 3D-CEUS.Enhancement patterns,sharpness or three-dimensional effect in different phases,definition of blood vessel in arterial phase and spatial relationships of tumor and vascularity of different hepatic tumors on the two imaging modes were compared statistically,the value of two modes in differential diagnosis of hepatic tumors was also evaluated.ResultsDynamic 3D-CEUS was similar to 2DCEUS with respect to enhancement patterns,sharpness or three-dimensional effect of tumor in arterial and portal phases( P ＞0.05).And there was no significant difference between the value of dynamic 3D-CEUS and 2D-CEUS in differential diagnosis of hepatic tumors:the sensitivity,specificity,positive predictive value and negative predictive value were 98.0％,81.3 ％,94.3 ％ and 92.9％ for 3D-CEUS,and were 96.1％,81.3％,94.2％ and 86.7％ for 2D-CEUS,respectively.However,3D-CEUS was superior to 2D-CEUS in the display of definition of blood vessel in arterial phase,and spatial relationships of hepatic tumor and vascularity( P ＜0.05).ConclusionsDynamic 3D-CEUS is a useful technique in the differential diagnosis of hepatic tumors as well as 2D-CEUS,and dynamic 3D-CEUS displays the spatial relationship of hepatic tumors and vascularity more visually and effectively,which provides additional information in the diagnosis and therapy of hepatic tumors.

Objective To evaluate the dynamic contrast-enhanced MRI (DCE-MRI)in detecting experimentally induced testicular ischemia. Methods Thirty healthy male New Zealand rabbits were randomly assigned into 6 groups. There were 5 rabbits in each of the following experimental groups: ( 1 ) Normal control, (2) Sham-operated, (3) ischemia of 3 h group, (4) ischemia of 6 h group, (5) ischemia of 12 h group, (6) ischemia of 24 h group. In all experiment groups, the right testis served as the internal control while the left testis served as the experimental side. DCE-MRI for each animal lasts about 10 minutes. Signal enhanced ratios (SERs) of ROI for both sides of each group were calculated by a computer, and parameters of SERs of 30 s, 75 s, 120 s and maximal SER were used for statistical analysis.Time intensity curves (TICs) were made for two sides of each group via Excel 2003 software and classified into 4 types. Statistical analysis was performed to compare the differences of SERs between left and right testis by two independent Kolmogorov-Smirnov test. Results In group I and 2, significant enhancement was observed on both testes of 10 rabbits. The enhancement decreased gradually with the elongation of ischemia in torsion groups. Three cases of type Ⅰ and 2 cases of type Ⅱ were observed in group 1,5 cases of type Ⅰ in group two, 2 cases of type Ⅰ and 3 cases of type Ⅱ b in group three, 2 cases of type Ⅰ and 2 cases of type Ⅱ b in group four, 5 cases of type Ⅱ b in group five and 5 cases of type Ⅲ in group six were noticed in the left testes. And in TICs of right testes, all cases showed TICs of type Ⅰ except 2 cases of type Ⅱ a in group six. In four torsion groups, the values for SER75 of the left side were 0. 084%, 0. 076%, 0.164% and 0.065%, while the right side were 0.255%, 0.410%, 0.586% and 0.302% (P ＜0.05). The values for SER120 in group three, five and six were 0.221% , 0.158% and 0.059% for the left side, and 0.405%,0.522% and 0.207% for the right side(P ＜0.05). The values for MSER in group three, five and six were 0.217% ,0.164% and 0.072% for the left side, and 0.405%, 0.586% and 0.302% for the right side(P ＜0.05). Conclusion DCE-MRI technique may be useful in the diagnosis of testicular torsion, which shows potential in the clinical application.

OBJECTIVETo investigate the relationship between polymorphisms of genes (CYP17, CYP19 and SULT1A1) involved in estrogen metabolism and susceptibility to breast cancer in Chinese women.

METHODSA case-control study was performed. PCR-base restriction fragment length polymorphism (PCR-RFLP) and short tandem repeat polymorphism (STRP) assays were used to detect the polymorphism distribution of CYP17, CYP19 and SULT1A1 in 213 breast cancer cases and 430 matched controls. Logistic regression analyses were used to determine the OR, multivariate adjusted OR and 95% CI of each and all three genes and estrogen exposure factors on the risk of breast cancer. Relationship between polymorphisms and clinic-pathological features was also assessed.

RESULTSThe frequency of A2 allele of CYP17 was 49.8% in cases and 49.1% in controls (P > 0.05). The frequency His allele of SULT1A1 in cases (13.6%) was significant higher than that of controls (9.5%) (P = 0.03). There was also significant difference in the frequencies of (TTTA)10 allele CYP19 which was 12.4% in cases and 8.2% in controls (P = 0.02). Multigenic model indicated that there was an increased risk of breast cancer with more numbers of high-risk genotypes in a dose-response effect (trend P = 0.05). Data from multivariate analysis showed that the allele of SULT1A1 His and CYP19 (TTTA)10 was positively associated with the risk of breast cancer. Other well-established risk factors as higher estrogen exposure including total years of menstrual, early menarche etc, and women with a higher BMI and WHR were all served as independent risks.

CONCLUSIONThis study indicated that the polymorphisms of estrogen-metabolizing genes were related to breast cancer.

Aromatase ; genetics ; Arylsulfotransferase ; genetics ; Breast Neoplasms ; genetics ; Case-Control Studies ; China ; Female ; Genetic Predisposition to Disease ; Humans ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Steroid 17-alpha-Hydroxylase ; genetics

Objective To investigate the optimal scheme about the external loading of resetting tensile stress for clavicular fracture,so as to provide some mechanical references for optimal design of external fixators for clavicular fracture.Methods The shoulder CT scanning data from a volunteer were used to establish the three-dimensional model of human left shoulder by Mimics software.In order to better simulate the actual human shoulder,the ligament tissues were added in the model and the constraints of muscles were considered,and the experimental model for simulation analysis on resetting tensile stress was established.By 3 simulation experiments,the impacts from directions of resetting tensile stress,positions of action points and value ranges were simulated and analyzed,respectively.The resetting tensile stresses obtained from clavicle of the shoulder model under various external loading cases were simulated by orthogonal test methods.Results When the θ (angle between the horizontal plane projection of shoulder force which was loaded by external fixator and the coronal axis) was greater than 45°,clavicle could obtain the resetting tensile stress,which became greater with the angle θ increasing.When η (angle between the sagittal plane projection of shoulder force which was loaded by external fixator and the sagittal axis) was 30°-45°,the resetting tensile stress of clavicle was the maximum.The farther the position on which the loads from external fixator was exerted from the horizontal position of clavicle,the larger the resetting tensile stress of clavicle was.Conclusions The results of orthogonal test show that the optimal resetting tensile stress is produced with parameter combination of θ =65°,η =50°,acting on the center of humerus head of the shoulder.

BACKGROUNDObstructive sleep apnea hypopnea syndrome (OSAHS) is regarded as a disease with strong genetic background and associated with hypoadiponectinemia. It is worthwhile to investigate the possible correlation between the single nucleotide polymorphisms (SNPs) in the adiponectin gene and OSAHS.

METHODSWith the TaqMan polymerase chain reaction (PCR) method, the SNPs at positions 45 and 276 in the adiponectin gene were determined in Chinese of Han nationality in Nanjing district consisting of 103 OSAHS patients (OSAHS group) and 67 normal controls (control group). The association of adiponectin genotype polymorphisms at positions 45 and 276 with OSAHS was analyzed.

RESULTSNo evidence of a direct association was found between OSAHS and adiponectin genotype SNP at positions 45 and 276 (P > 0.05). However, compared with those OSAHS patients having G/T + T/T genotype at position 276, the OSAHS patients with G/G genotype showed a longer neck circumference, a prolonged duration of the longest apnea event, and an elevated level of blood cholesterol and low-density lipoprotein cholesterol (P < 0.05).

CONCLUSIONSNo direct association was suggested between OSAHS and adiponectin genotype distribution at positions 45 and 276 in Chinese of Han nationality in Nanjing district. However, in OSAHS patients, those with adiponectin G/G genotype at position 276, seemed to have a higher potential risk in development of OSAHS than those having adiponectin SNP276 G/T + T/T genotype.

Adiponectin ; genetics ; Adult ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Polymorphism, Single Nucleotide ; genetics ; Sleep Apnea, Obstructive ; genetics

BACKGROUNDEndogenous estrogen plays a very important role in the carcinogenesis and progression of breast cancer. The enzymes involved in the biosynthesis and metabolism of estrogen have been proposed to contribute to this effect. To examine this hypothesis, we conducted a case-control study to investigate the relationship between polymorphisms of genes responsible for estrogen biosynthesis (CYP17, cytochrome P450c17a and CYP19, aromatase cytochrome P450) and estrogen sulfation of inactivation (SULT1A1, sulfotransferase1A1) and the risk of breast cancer in Chinese women.

METHODSThis study involved 213 breast cancer patients and 430 matched controls. PCR-based restriction fragment length polymorphism (RFLP) and short tandem repeat polymorphism (STRP) assays were used to detect the mononucleotide transition of CYP17 and SULT1A1 and tandem repeat polymorphism of CYP19. Logistic regression analyses were used to determine OR and 95% CI of each and all three high-risk genotypes, of all three genotypes combined, and of estrogen exposure factors. The relationship between each high-risk genotype and clinicalpathological characteristics were also assessed.

RESULTSThe frequency of A2 allele of CYP17 was 49.8% in cases and 49.1% in controls (P = 0.82). The frequency of His allele of SULT1A1 was significantly higher in cases (13.6%) than in controls (9.5%) (P < 0.05). There was also significant difference of the (TTTA) 10 allele of CYP19 which was 12.4% in cases and 8.2% in controls (P < 0.05). When the CYP17 A2 allele, CYP19 (TTTA) 10 and SULT1A1 His allele were considered as the "putative high-risk" genotype, there was an increased risk of breast cancer with the number of high-risk genotypes in a dose-response effect (trend, P = 0.05). In multivariate analysis, the SULT1A1 genotype remained the most significant determinant for breast cancer, with OR = 2.37 (95% CI 1.23-4.74), followed by CYP19, with OR = 1.75 (95% CI 1.27-3.56). The (TTTA) 10 allele of CYP19 was associated with tumor size, and the His allele of SULT1A1 associated with status of lymph node metastasis.

CONCLUSIONSThis study supports the hypothesis that breast cancer can be initiated by estrogen exposure and that estrogen metabolizing genes are involved in this mechanism. This multigenic model is useful for identifying individuals who are at higher risks of breast cancer.

Adult ; Aged ; Aromatase ; genetics ; Arylsulfotransferase ; genetics ; Breast Neoplasms ; etiology ; genetics ; Case-Control Studies ; Estrogens ; metabolism ; Female ; Genetic Predisposition to Disease ; Humans ; Middle Aged ; Polymorphism, Genetic ; Risk Factors ; Steroid 17-alpha-Hydroxylase ; genetics

OBJECTIVETo identify the relationship between coagulation factor V (FV) gene single nucleotide polymorphisms (SNPs) and venous thromboembolism (VTE).

METHODSThe FV clotting activity (FV: C) and FV antigen (FV: Ag) in plasma of VTE group (111 patients) and normal control (110 patients) were detected using one-stage clotting assay and ELISA, respectively. Five pairs of primers of the F V polymorphisms including Asp79His, Arg306The, Arg306Gly, Arg506Gln and Ile359The/His1299 Arg were synthesized and amplified by PCR. The PCR products were digested by restriction enzyme using PCR-RFLP. The detected polymorphisms were confirmed by direct sequencing. The samples containing the polymorphisms were screened for coding regions of all F V exons with direct sequencing.

RESULTSThe plasma levels of F V: C and F V: Ag of VTE group and normal control were (106.9 +/- 28.0)%, (110.4 +/- 33.3)% and (102.4 +/- 30.9)%, (102.1 +/- 24.1)%, respectively. The plasma level of FV: Ag was significantly different between VTE group and normal control. However, there was no difference in F V: C levels. Polymorphisms for the fore mentioned 5 primer pairs were not found in either patients or normal controls. Polymorphism of His1299Arg was identified in 5 patients with VTE and 3 normal controls. And these 5 cases also combined Met1736Val polymorphism, 3 of them combined another Asp2194Gly polymorphism.

CONCLUSIONThe higher plasma level of F V: Ag contribute to venous thromboembolism. There is no relationship between polymorphisms of Asp79His, Arg306The, Arg306Gly, Arg506Gln, Ile359The and venous thromboembolism in Chinese studied. Polymorphism His1299Arg is higher in VTE group than in normal control, but has no statistical difference. Polymorphisms of His1299Arg, Met1736Val and Asp2194Gly are linked disequilibrium in Chinese Han population.

Factor V ; genetics ; Female ; Gene Frequency ; Humans ; Linkage Disequilibrium ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Venous Thromboembolism ; genetics