Type 1 diabetes is a autoimmune disease that occurs under the influence of both genetic predisposition and environmental factors,mediated mainly by T lymphocytes with various kinds of other involved immune cells.The autoimmune attacks eventually lead to the islet beta cell destruction and insulin insufficiency.Multiple fundamental studies and clinical trials have revealed that umbilical cord blood pluripotent stem cells have the potential to help restore immune balance,induce immune tolerance,stop the autoimmune attacks against beta cells,and promote beta cell regeneration in type 1 diabetes mellitus (T1DM),by means of cell to cell contact and soluble cytokine secretion.For the past few years,the National Clinical Research Center for Metabolic Diseases of Second Xiangya Hospital has been leading the research on stem cell education therapy and has performed the therapy for 35 juvenile type 1 diabetes patients from China and foreign countries,introducing a novel treatment for T1DM.

The duck circovirus (DuCV) infection in sick ducks from Fujian Province was investigated. The liver samples of 43 sick Muscovy ducks with infectious serositis were collected from 12 duck farms in Fujian Province.Based on the published sequences of DuCV, two primers were designed for the detection of DuCV and four pairs of primers were designed to amplify four overlapping fragments that cover the complete genome of DuCV. The specific PCR products were amplified from positive samples. The fragments were then cloned into pMD18-T vector and sequenced, and the full length genomic sequence of the FJ0601 isolate of DuCV was obtained. PCR analysis showed that the proportion of ducks which were positive for circovirus was 79% and 10 out of the 12 farms were positive. Sequence analysis showed that the complete genome of DuCV-FJ0601 was 1988 bp and possessed features common to the family Circoviridae which included a stem-loop structure and the Rep protein motifs. Homology analysis showed that FJ0601 isolate of DuCV had 97.3%～97.5% nucleotide sequence identity to all the four Taiwan isolates (TC1/2002, TC2/2002, TC3/2002, TC4/2002), 82.9% identity to the America (33753-52) isolate and 82.3% identity to the Germany isolate. Phylogenetic analysis with Clustal W, however,showed that FJ0601 isolate of DuCV was on a common branch with Taiwan isolates, and Germany and America isolates belonged to the other branch.

Objective:To establish a method for the detection of 8 single nucleotide polymorphisms (SNPs) of RAC1 gene,and to analyze the genotypic and allelic distributions of the 8 SNPs in healthy Chinese Han population.Methods:The real-time fluorescence TaqMan-MGB probes allele classification technology was used for the determination of the 8 SNPs in 150 cases of healthy volunteers in Hubei China ,and the results were validated by direct gene sequencing.Results: The method established in this study can accurately screen the genotypes of the 8 SNPs of human RAC1 gene.Highly linkage disequilibriums were found between rs 10951982 and rs9374 , rs702482 and 836488 ,respectively.All the 8 sits were in accordance with Hardy-Weinberg equilibrium.Six Tag-SNPs were selected by Heploview software:rs10951982 ,rs6954996 ,rs6951997 ,r1s2977 ,rs702482 and rs702483.The MAFs of the 8 SNPs in our study were close to the MAFs in CHB and Asian in Hap Map database .Significant distribution differences were also observed in different races.Conclusion:No significant differences are observed in this study in healthy Chinese Han population.But differences are found when compared with the data of other races in Hap Map.

Objective:To develop a method for the detection of 10 single nucleotide polymorphisms(SNPs) of Nm23 gene,and to explore the genotypic and allelic distributions of the 10 SNPs in Chinese Hans population in Wuhan.Methods: Two hundreds healthy subjects ,115 men and 85 women included ,were enrolled as DNA sample donors.The real time TaqMan-MGB genotyping assay was used for the determination of the 10 SNPs selected ,and the results were validated by direct gene sequencing.Results:The method established could accurately and quickly screen the genotypes of the 10 SNPs of Nm23/NDPK gene.Distribution frequencies of the 10 SNPs were similar to these in other researches as well as these of HCB.All the loci follow the Hardy-Weinberg equilibrium.Highly linkage disequilibriums were found between rs 16949649 and rs7207370 , rs16949649 and rs34214448 , rs2159359 and rs2302254 , rs2159359 and rs8075231 ,rs2159359 and rs2041296 ,as well as rs2159359 and rs8071647 ,respectively.Four Tag SNPs:rs34214448 , rs2302254 ,rs11868380 and rs2318785 were initially selected by Heploview software.Conclusion:The method established for SNP gen-otyping can meet the needs for rapid analysis of Nm 23 gene polymorphisms ,and may have great values in investigating the association between gene polymorphisms and diseases as well as adverse drug reactions.

Digital X-ray imaging technique has been developing rapidly in recent years, and makes it possible for people to be radiated by lower X-ray dose than before to get the images for diagnosis. Current development in 7 aspects about digital roentgenography is introduced in this article.

Objective:To study the association of RAC1 gene polymorphisms with protein expression levels of Rac1-GTP. Methods:A total of 182 healthy Hans population in Hubei were recruited. The 4 tag-SNPs in RAC1 gene were genotyped by Real time TaqMan-MGB genotyping assay. And the Rac1-GTP protein levels in plasma samples from all participants were determined by enzyme linked immunosorbent assays ( ELISA ) . The distribution characteristics of Rac1-GTP expression levels were also analyzed. Furthermore,the expression levels of Rac1-GTP were compared among different genotypes of the 4 tag-SNPs in RAC1 gene. Results:The distribution of Rac1-GTP expression levels was positive skewed in healthy Chinese Hans population. The expression levels were significantly higher in females than in males (P<0. 05),and appeared in decreased trend with age,but without significant differences (P>0. 05). Different expression levels of Rac1-GTP were observed in different genotypes for rs702482 and rs10951982 (P<0. 05). However,no significant difference was found for rs702483 and rs6954996 (P>0. 05). Conclusion:RAC1 genetic polymorphisms can potentially affect the expression levels of Rac1-GTP protein in healthy Chinese Hans population.

Objective To investigate the diagnostic value and imaging characteristics of MRI in the placenta accreta of patients with retained placenta.Methods Archieves of patients with retained placenta confirmed by clinical pathology were reviewed,with focus on imaging findings of MRI,including location of and signal intensity of placenta attach,hematometra,presence and absence of placenta accreta,signal intensity of retained placenta,shape,margin and border of retained placenta.The imaging characteristics were analysized with blindness and compared between patients with and without placenta accreta.Diagnostic value of MRI and ultrasound for the placenta accreta of patients with retained products of conception was assessed.Results There was no significant difference between the two groups of MRI characteristics except that the edge of the placenta was not clear.The sensitivity,specificity,accuracy,positive predictive value and negative predictive value for the evaluation of placenta accreta by MRI and ultrasound were 93.8% vs 56.3%,90.9% vs 90.9%,92.6%vs 70.4%,0.94 vs 0.9,0.91 vs 0.59,respectively;there were significant differences between MRI and ultrasound for sensitivity, specificity and negative predictive value.Conclusion M RI is substantially better than ultrasound for the diagnosis of placenta accreta. The unclear placenta edge and signs of placental infiltration of the myometrium are the basis for MRI diagnosis of placenta accreta.

Objective Adhesion or too highly located folds upper eyelid and even hlepharoptosis are common complications after double eyelid operation. The method was designed to correct these de-formities. Methods The incision of upper eyelid was designed under the scar. Skin was resected between the former and the new double line, if there was excessive skin. Completely relieve the adhesion until nor-mal tissue was exposed. Expose lateral extension of retro-orbicularis oculi fat pad. An upper pedicle fat flap was formed after cutting tissues off along bilateral borders. Appropriately and transversely cutting lateral portion of the pedicle made it rotate inside downward with enough blood supply. The fat flap was fixed with released orbital fat and the lower edge of fat flat fixed with upper border of supratarsal to restorate the volume. For severe condition, fat granule injection could be used after the skin was sutured. Results 42 cases (80 eyes) were treated in the past two years. The deformities of 32 cases were corrected by the application of retro-orbicularis oculi fat pad. Fat granule injection was applied for the other 10 pa-tients meanwhile. 30 cases (60 eyes) were followed up for 6 to 24 months. The results were satisfactory. Conclusion The methods are performed easily and the results are satisfied. It is worth of recommenda-tion.

Objective:To investigate whether hepatitis B virus X protein (HBx) mediates the podocyte injury through reactive oxygen species (ROS) /nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) signaling pathway.Methods:HBx-overexpressing lentivirus was transfected into renal podocytes of mouse to mimic the pathogenesis of hepatitis B virus-associated glomerulonephritis. Podocytes were divided into the following five groups: blank control group (no special treatment), negative control group (transfected with control lentivirus), HBx overexpression group (transfected with HBx overexpression lentivirus), HBx overexpression+NLRP3 siRNA group (transfected with HBx overexpression lentivirus and NLRP3 siRNA), and HBx overexpression+ROS inhibitor group (transfected with HBx overexpression lentivirus and adding ROS inhibitor). The morphological changes of podocytes were observed with electron microscope. The generation of ROS was detected by dichlorodihydrofluorescein diacetate assay (DCFH-DA). Hoechst 33342 staining was used to observe the morphological and quantitative changes of podocyte nuclei. Enzyme-linked immunosorbent assay was used to detect caspase-1 activity, and the levels of lactate dehydrogenase, interleukin (IL)-1β and IL-18. Quantitative real-time polymerase chain reaction and Western blotting were used to detect the expression levels of mRNA and protein of pyroptosis-related protein, such as NLRP3, apoptosis-associated speck-like protein containing card (ASC), caspase-1, IL-1β and IL-18. TUNEL staining and flow cytometer were used to detect the number of pyroptosis cells. Immunofluorescence staining was used to detect the expression levels of desmin and nephrin.Results:After successful infection of podocytes with HBx-overexpressing lentivirus, pyroptosis-related morphological changes in the cells were observed under electron microscope. The level of ROS in the HBx overexpression group was significantly higher compared to the negative control group ( P<0.05). Hoechst 33342 staining revealed condensed nuclei in the HBx overexpression group. TUNEL staining and flow cytometer demonstrated that podocytes underwent increased pyroptosis in the HBx overexpression group. The mRNA and protein expression levels of pyroptosis-related proteins such as NLRP3, ASC, caspase-1, IL-1β and IL-18 were up-regulated upon HBx overexpression (all P<0.05). Caspase-1 enzyme activity, lactate dehydrogenase and desmin expression levels were enhanced after HBx overexpression (all P<0.05). However, NLRP3 knockdown or addition of ROS inhibitors attenuated the pyroptosis and increased expression levels of pyroptosis-related proteins caused by HBx overexpression (all P<0.05). Conclusion:ROS/NLRP3 pathway plays an important role in HBx-induced podocyte pyroptosis.

Objective To explore the biomechanical responses of the cupula of the human semicircular canal to three basic rotational perception processes.Methods A one-dimensional visual semicircular canal model was successfully fabricated using three-dimensional printing and hydrogel physical cross-linking technologies,and the response deformation of the cupula was explored by applying constant angular velocity,constant angular acceleration,and sinusoidal oscillation stimulations.Results The time constant of the biomimetic semicircular canal model was stable at approximately 3 s and close to the human time constant.The displacement deformation of the ampullary cupula was proportional to the angular acceleration applied.Under sinusoidal oscillation stimulation of 0.07-5.00 Hz,the gain of the semicircular canal increased from 1.54 um/° rises to 42.34 μm/°,but the phase difference decreased from 109.72° to 11.27°.Conclusions The biomimetic semicircular canal model prepared in this study can accurately simulate the working mechanism of the human semicircular canal and is expected to play a role in mechanism research and disease diagnosis of the human vestibular semicircular canal.