The endothelium of 20 cases of rabbit cornea including the normal and postoperated ones (2 weeks and I, 2, 3 months after penetrating keratoplasty) were observed by scanning electron microscope. The photographs of the endothelium in graft-host junction were analysed by computer-assisted image analysis system and the morphometric indexes are as follows: area of the cells, perimeters, density, figure coefficient, long axis, coefficient of variation of the area, and others. After operation the morphology and the density of the endothelial cells were different from the normal ones obviously, but the cells changed from enlargement and irregularity to normal progressively. At the time when the corneas were operated for 3 months, some differences still existed.

Objective To evaluate left ventricular systolic synchronicity in patients with dilated cardiomyopathy by V-plane imaging and compare with clinical commonly used tissue Doppler imaging (TDI),evaluate the relevance and consistency between these two parameters.Methods 20 patients diagnosed with DCM and 20 healthy controls were enrolled,2D images,V-plane imaging and TDI waveform were acquired.Time to peak velocity of left ventricular 12 segments were measured by TDI and the standard deviation (TDI_SD) were calculated.Displacement time to peak were measured by V-plane and the standard deviation (V-plane_SD) were calculated.Results Compared with control group,TDI_SD and V-plane_SD increased significantly (P ＜ 0.01),TDI time to peak increased and V-plane time to peak decreased significantly(P ＜0.01).In the two group,12 segment time to peak measured by V-plane were significantly higher than TDI(P ＜ 0.01).There was a significant correlation between TDI_SD and V-plane_SD (r =0.925,P ＜0.001).Bland-Altman plot showed that 95％ plots of ratio of TDI_SD and V-plane_SD were among limits of agreement,which was (0.50,1.36).Conclusions Left ventricular systolic synchronicity in DCM patients can be observed by V-plane imaging.V-plane has significant relevance and consistency with TDI,and can overcome the limitations of TDI that 12 segments can not be displayed in the same cardiac cycle.

Objective To construct HaCaT cell lines stably expressing the wild type human GJB6 gene or its mutant by using a Tet-On lentiviral vector, and to lay an experimental foundation for studies on pathogenesis of hidrotic ectodermal dysplasia. Methods The wild-type human GJB6 gene and its mutant (A88V)were amplified by PCR, and then inserted into the Tet-on lentivirus plasmid to construct recombinant lentivirus vectors. The recombinants were identified by gene sequencing and enzymatic digestion. Cultured HaCaT cells were classified into three groups to be transfected with a negative control lentiviral vector (NC group), the lentivirus vector expressing the wild-type human GJB6 gene (WT group), or the lentivirus vector expressing the mutant human GJB6 gene (MU group). Puromycin was used to select HaCaT cell clones stably expressing the GJB6 gene which encodes the connexin 30 (Cx30)protein. The selected HaCaT cell clones were cultured with or without tetracycline for 48 hours, thereafter, real-time PCR(RT-PCR) was performed to detect GJB6 gene mRNA expression, Western-blot analysis to measure expressions of Cx30 and FLAG-tag proteins, and cell counting kit 8 (CCK8)assay to evaluate cellular proliferative activity. Results Enzymatic digestion and gene sequencing showed that recombinant lentivirus plasmids were successfully constructed. RT-PCR showed evidently increased mRNA expression of the GJB6 gene in stably transfected HaCaT cells. Moreover, the expression abundance of the GJB6 gene was 112.369 times higher in the WT group induced by tetracycline than in that without tetracycline treatment (P < 0.05), and 2.249 times higher in the MU group induced by tetracycline than in that without tetracycline treatment (P < 0.05). Western-blot analysis showed that Cx30 and FLAG-tag proteins were stably expressed in the WT group and MU group after induction with tetracycline, while neither of them was observed in the WT group or MU group without tetracycline treatment, or in the NC group. Significant differences were noted in cellular proliferative activity (expressed as the absorbance value at 450 nm)between the MU group with and without tetracycline treatment and between the WT group with and without tetracycline treatment at 4, 8, 12, 24, 36 and 48 hours (all P <0.05), but not between the NC group with and without tetracycline treatment at any of the above time points (all P >0.05). Conclusion HaCaT cell lines which stably express the wild-type GJB6 gene or its mutant(A88V)are successfully constructed.

Objective To target the disease gene of disseminated superficial form of porokeratosis (DSP) in a six-generation of a Chinese family including a total of 254 family members in Shandong province. Methods The clinical data and the peripheral blood samples were collected in the pedigree members. The genomic DNA was extracted from the blood samples. A genome-wide scan was performed using 382 pairs of primers labelled with fluorescent stain. The primers were designed for human autosomes. The sequencing results were analyzed by the software of Genescan and Genotype. Linkage analysis was processed by Linkage software package to define the region of disease gene. For fine targeting the disease gene, other 10 micro-satellite markers for the above region were set up for further fine sequencing. Results We obtained the maximum two-point LOD scores of 3.06 at micro-satellite marker D12S78 (recombination fraction ? = 0.00). After fine mapping, the DSP gene is located within a 38.5 cM region between markers D12S326 and D12S79. Conclusion The DSP gene is mapped to chromosome 12q21.2~24.2.

Objective To create a three-dimensional speckle tracking imaging (3DSTI) multiparameter analysis model to improve the diagnostic efficiency of obstructive coronary artery stenosis.Methods One hundred and four patients with chest pain were divided into two groups:coronary heart disease(CHD) group (61 patients) and control group (43 patients) according to the result of selective coronary angiography (SCA).The two groups' clinical data and echocardiographic parameters were aquired,including mitral flow E and A velocities,E peak deceleration time,isovolumic relaxation time (IVRT),mitral annulus velocity e' and a' peak in diastolic and s' peak in systolic by tissue Doppler imaging (TDI),and left ventricular ejection fraction (LVEF).The global longitudinal peak strain (2D-GLPS) by two dimensional speckle tracking,global longitudinal peak strain (3D-GLPS),circumferential peak strain(3D-GCPS),radial peak strain (3D-GRPS) and area peak strain (3D-GAPS) were acquired.Results For conventional parameters,there were no significant difference between the two groups.Compared with control group,TDI e' peak,3D-GRPS decreased,significantly E/e',2D-GLPS,3D-GLPS,3D-GAPS,3D-GCPS increased significantly (P ＜0.01).For single parameter,area under the ROC curve (AUC) were successively 3D-GAPS(0.766) ＞ 3D-GLPS(0.746) ＞ 2D-GLPS(0.746) ＞3D-GRPS(0.727) ＞ s' (0.703) ＞E/e' (0.688)＞3D-GCPS(0.686).AUC for single and multi technology were successively p-Union(0.856)＞p-3DSTI(0.772) ＞ p-TDI (0.757) ＞ p-2DSTI (0.746).Conclusions 3DSTI together with multi-parameter analysis model can significantly improve the diagnostic efficiency of obstructive coronary artery stenosis.Area strain is an independent predictor of obstructive coronary artery stenosis.

Objective To investigate the treatment values of precise target delineation of chest MRI for lung cancer Methods From August 2011 to February 2015 , 45 non-small cell lung cancer patients were given chest CT scans and MRI scans before radiotherapy , and then active target tumor delineation , then related influencing factors were analyzed. Results All patients completed CT scans and MRI positioning. For patients that it was difficult to identify lung tissue lesions caused by lung cancer through CT , their MRI imaging showed high signal and the boundaries between the tumor and surrounding normal tissue became relatively clear. Meanwhile , 20 patients of borders were diagnosed by CT , while 25 by MRI; 36 patients with lymph node metastasis were diagnosed by CT while 40 by MRI. The difference was statistically significant (P<0.05). Univariate logistic regression analysis showed that pathological type and atelectasis were the influence factors for CT and MRI tumor target delineation differences (P<0.05), and multivariate logistic regression analysis showed the atelectasis was the main factor (P<0.05). Conclusion Compared with CT, breast MRI can precisely delineate target to improve the accuracy of target localization before radiotherapy. It can help determine lymph node metastasis and avoid the impact of atelectasis then ensure the accuracy of radiotherapy.

ObjectiveTo detect the mutation of GJB2 gene in a Chinese family with Vohwinkel syndrome.MethodsClinical data were collected from 5 patients with Vohwinkel syndrome in a family,and blood samples were obtained from the 5 patients and 4 unaffected individuals in the family as well as from 100 normal human controls.Genomic DNA was extracted and subjected to PCR for the amplification of the entire encoding and flanking sequences of GJB2 gene(1015 bp) followed by bidirectional sequencing with the ABI PRISM 3730 automatic DNA sequencer.Finally,sequence alignment was carried out by using the software Sequencher 4.10.1 Demo.ResultsA heterozygous missense mutation 196G→C in the GJB2 gene,which resulted in the substitution of aspartic acid by histidine at codon 66 (D66H) in the first extracellular domain of the protein,was observed in all the patients of this family,but in none of the 4 unaffected individuals in this family or the 100 normal human controls.ConclusionThe D66H missense mutation in the GJB2 gene may contribute to the occurrence of Vohwinkel syndrome in Chinese Han population.

Objective To confirm the diagnosis and to localize the pathogenic gene of ectodermal dysplasia in a family SUffering from only hair and nail abnormalities.MethodsBlood samples were collected from 7 affected patients and 15 unafiected individuals in the family.Genomic DNA was extracted from blood samples by routine phenol-chloroform methods.The whole coding regions of candidate genes K16,K17,K6a,K6b and GJB6 were amplified by PCR followed by direct sequencing.Then,the gene mutation was further confirmed at mRNA level by RT-PCR.ResultsA heterozygous missense mutation 3 1G→A in the GJB6 gene.which leads to the substitution of glycine by arginine at codon 11(G11R)on the N-terminal of the protein,was detected in all the patients.but in none of the 15 normal individuals in this family.The mutation was also confirmed in the CDNA originating from the proband's skin biopsy.Conelusionn A missense mutation G31A.which has been shown previously to cause hidrotic ectodermal dysplasia(HED),is localized in the GJB6 gene of patients in this family.

ObjectiveTo analyze the quality changes of Platycladi Semen before and after the deterioration of moth-eaten and rancidity during storage. MethodFour types samples of Platycladi Semen， including normal， moth-eaten， oxidative rancidity and hydrolytic rancidity， were determined for volatile components， odor， and taste based on headspace solid phase microextraction/gas chromatography-mass spectrometry （HS-SPME/GC-MS） and electronic sensory techniques such as electronic nose and electronic tongue. Volatile components were identified by searching the database and manual comparison， the odor and taste were determined by the response values of the electronic nose and electronic tongue sensors， and the difference between samples before and after deterioration was studied by multivariate statistical analysis. ResultA total of 85 compounds were identified in Platycladi Semen samples. Compared with the normal samples， the number of volatile compounds in samples after hydrolytic rancidity decreased by 5， the number of volatile compounds in samples after moth-eaten and oxidative rancidity increased by 1 and 21， respectively. Aldehydes and acids accounted for majority of types. Among them， the contents of N-hexanoic acid， hexanal and propionic acid in the samples of oxidative rancidity reached 11.49%， 10.21% and 7.52%， which became the key indicators of rancidity. There was significant variance among the odor components corresponding to W1W， W2W and W1S sensors by electronic nose analysis. It was indicated that the value of sourness in deteriorated samples generally increased by mean of electronic tongue analysis. Compared with normal samples， the moth-eaten samples had changed slightly and rancidity samples had changed significantly especially oxidative rancidity samples of volatile components， odor and taste by multivariate statistical analysis. ConclusionIn terms of Platycladi Semen， the oxidative rancidity caused by nature storage for 12 months has the greatest impact on the quality. Therefore， it should be mainly to prevent oxidative rancidity to ensure the quality of Platycladi Semen.