Objective To explore the characteristics of proinsulin secretion in latent autoimmune diabetes in adults (LADA). Methods Fasting and 2 h sera in oral glucose tolerance test from 36 LADA patients, 37 type 2 diabetic patients and 43 healthy controls were collected to test glucose, proinsulin (PI) and C-peptide (CP) by radioimmune assay. Glutamie acid decarboxylase antibodies (GAD-Ab) were determined by radioligand assay.Results (1) Fasting proinsulin (FPI) and 2 h proinsulin (PPI) level in LADA patients were lower than those in type 2 diabetic patients (P<0.05), being both significantly inereasad compared with healthy controls (P<0.05 or P<0.01); The ratios of FPI/FCP and PPI/PCP (%) in LADA were beth significantly higer than those of type 2 diabetes mellitus and controls (P<0.05 or P<0.01); (2) LADA type-1 (GAD-Abe>0.3) patients showed lower PI levels(P<0.05 or P<0.01) and higher PI/CP ratio (all P<0.05) than LADA type-2 (0.05≤GAD-Ab<0.3); Meanwhile, there were no significant differences in above parameters between LADA type-2 and type 2 diabetes meUitus (P>0.05). (3) GAD-Ab index was negatively correlated with FPI and PPI in LADA group (r=-0.236 and-0.268, both P<0.05), and positively correlated with PPI/PCP (r=0.254, P=0.030).Meanwhile BMI was positively correlated with FPI, PPI and PI/CP in type 2 diabetes mellitus (all P<0.01). No factor entered the multiple regression analysis for predieting the hyperproinsulinemia and dispropriately elevated proinsulin levels in LADA patients. (4) According to the 99.5 th percentile of proinsulinemia in the healthy controls, which is defined as the cutoff point dispropriately elevated proinsulin levels, the proportion of subjects with fasting dispropriately elevated proinsulin levels (FPI/FCP) were 77.8%, 62.2% and 2.3% in LADA, type 2 diabetes meUitus and controls respectively, and PPI/PCP 83.3%, 51.4% and 2.3% respectively. Conclusion LADA patients, as well as type 2 diabetic patients, all showed hyperproinsulinemia and disproportionately elevated proiasulin levels that were one of characteristics of defective β-cell function. Moreover, disproportionately elevated nproinsulin level is more evident in LADA patients than that in type 2 diabetics and this may be related to humoral immunity.

OBJECTIVETo construct plant expression pCAMBIA1301-PMI by substituting PMI for hygromycin resistance gene in pCAMBIA1301 and obtain transgenic Salvia miltiorrhiza f. alba using PMI-mannose selection system.

METHODThe 6-phosphomannose isomerase gene (PMI) of Escherichia coli was amplified by PCR. Sequence analysis showed that it shared 100% amino acids identities with the sequences of PMI genes isolates reported in the NCBI. Based on pCAMBIA1305, the plant expression pCAMBIA1305-PMI was constructed successfully by substituting PMI for hygromycin resistance gene in pCAMBIA1305. pCAMBIA1305-PMI was transformed into Agrobacterium tumefaciens LBA4404, and then the leaves of S. miltiorrhiza f. alba were inoculated in LBA4404 with pCAMBIA1305-PMI.

RESULTPlant expression pCAMBIA1301-PMI was successfully constructed and the leaves of S. miltiorrhiza f. alba inoculated in LBA4404 with pCAMBIA1305-PMI were selected on medium supplemented with a combination of 20 g x L(-1) mannose and 10 g x L(-1) sucrose as a carbon source. The transformation efficiency rate was 23.7%.

CONCLUSIONGenetic transformation was confirmed by PCR, indicating that a new method for obtaining transgenic S. miltiorrhiza f. alba plants was developed using PMI-mannose selection system.

Anti-Bacterial Agents ; pharmacology ; Biomarkers ; Cinnamates ; pharmacology ; Escherichia coli ; enzymology ; genetics ; Escherichia coli Proteins ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Hygromycin B ; analogs & derivatives ; pharmacology ; Mannose-6-Phosphate Isomerase ; genetics ; metabolism ; Plants, Genetically Modified ; drug effects ; genetics ; metabolism ; Salvia miltiorrhiza ; drug effects ; genetics ; metabolism ; Transformation, Genetic

Objective To evaluate the association of islet autoantibodies [ glutamic acid decarboxylase antibody(GADA),protein tyrosine phosphatase antibody(IA-2A)and insulin autoantibodies(IAA)1 with HLA- DQ genotypes in the first-degree relatives of autoimmune type 1 diabetes mellitus.Methods This was a cross- sectional and case-control study.Three hundred and fifty-one first-degree relatives with normal glucose tolerance of patients with type 1 diabetes mellitus and 376 healthy controls were recruited and measured for GADA,IA-2A and IAA by radioligand assay,and 156 first-degree relatives of patients with autoimmune type 1 diabetes mellitus and 278 controls were typed for genetic polymorphisms of HLA-DQ with PCR sequencing-based typing method.Results (1)DQA1*03,DQBI*0303,*0401 alleles and DQA1 * 03-DQBI * 0303,DQA1 * 05-DQBI * 0201,DQA1 * 03-DQBI * 0401 haplotypes were significantly increased in the first-degree relatives of autoimmune type 1 diabetes mellitus(P

The HLA system was discovered by virtue of the fact that it was polymorphic. The impetus for its discovery was the search for polymorphic antigens to match for transplantation, by analogy with the human red cell blood groups. The most usually DNA method of HLA typing is sequence specific oligonucleotides (SSO) and PCR sequence specific primers (SSP). SSO technique is perfectly suited for analyzing large number of samples, it is not suitable for individual or small numbers. The SSP method is ideal for typing individual samples, but it is costly and requires high capacity thermal cycles for larger numbers of samples. To set up a simple, quick, cheap and high resolution DNA method, were collected sixty-three cord blood samples from Guangzhou Cord Blood Bank, got DNA from blood by the traditional guanidine hydrochloride distillation method. Each sample was simultaneously typed by SSOP, PCR-SSP and reverse dot-blot hybridization (RDB) methods. All of typed is success. The results of three DNA methods are consistent each other. 60 HLA-DRB1 alleles could be accurately distinguished with the RDB method. Our results show that RDB method is a simple, quick, cheap and high resolution method for HLA-DRB types. It can be used in any HLA typing.
Fetal Blood ; immunology ; metabolism ; Genotype ; HLA-DR Antigens ; genetics ; Histocompatibility Testing ; methods ; Humans ; Nucleic Acid Hybridization ; methods

To verify that the circular forms of bocavirus genome exist in their host, bocavirus episomes were detected in fecal samples of healthy piglets using a semi-nested PCR method. Two species of porcine bocaviruses (PBoVG2-episome and PBoVG3-episome) were identified for the first time. The relevant terminal sequences of the noncoding region (405 and 511 nt, respectively) were also obtained. Sequence analyses and secondary structure prediction indicated that the PBoVG2-episome was more similar to that of human bocavirus 3 (HBoV3) but the PBoVG3-episome was quite different from that of other members of the genus Bocavirus. Discovery of episomal forms of porcine bocaviruses (PBoV) suggested that PBoV, like HBoV, used a different replication mechanism from other parvoviruses. The sequencing of episome Inverted Terminal Repeats (ITRs) also contributes to a possible alternative strategy for constructing infectious molecular clones of bocavirus in a future study.
Animals ; Base Sequence ; Bocavirus ; genetics ; physiology ; DNA, Circular ; genetics ; DNA, Viral ; genetics ; Genome, Viral ; genetics ; Molecular Sequence Data ; Polymerase Chain Reaction ; Swine ; virology

Homology of three WSSV isolates, which were sampled from r epresentative maritime space of China: Tanghai isolate (Bo Bay of China), Ningbo isolate (East China Sea), Shenzhen isolate (South China Sea) was compared. Both of the genome RFLP patterns and the characteristic structural proteins SDS-PAG E electrophero grams showed that they were quite same. It suggested that they were the same ki nd of WSSV virus that caused explosive epidemic diseases of shrimps (EEDS) throu ghout southern and northern China. The same large PCR products achieved when usi ng the PCR primers from RV－PJ＝PRDV (P. japonicus, Japan) and WSBV＝PmNOBII I(P.monodon Taiwan, China) respectively to amplify the genome from P.chine nsis (Tanghai, China) with high fidelity Taq Polymerase. The sequence identiti es of WSSV from P. chinensis with those from RV－PJ＝PRDV (P.japonicus, Japan) and WSBV＝PmNOBIII (P.monodon Taiwan, China) are 97% and 100% respect ively, the results provided additional evidence that WSSV reported in different parts of the Asian and Pacific regions maybe quite the same or just different va riants of the same virus.

OBJECTIVETo investigate the anti-inflammatory effect of bone marrow stromal cells (MSCs) transfected with recombinant adenovirus-mediated ciliary neurotrophic factor (CNTF) gene in C57BL/6 mice with experimental allergic encephalomyelitis (EAE).

METHODSAn adenovirus vector containing CNTF gene Ad-CNTF-IRES-GFP was constructed and transfected in the MSCs (MSC-CNTF). After examination of CNTF expression, the transfected cells were transplanted in C57BL/6 mice with MOG 35-55-induced EAE, which were monitored for the changes in the symptoms scores. The levels of tumor necrosis factor-alpha (TNF-alpha), inteferon-gamma (IFN-gamma), interleukin-12P35 (IL-12P35), and IL-10 in the peripheral blood of the mice were detected, and the number of MSC-CNTF cells in the spleen and spinal cord was counted. CD3+ T cell infiltration and TNF-alpha and IFN-gamma expressions in the lesions were also observed after the cell transplantation.

RESULTSCNTF gene transfection resulted in significantly increased CNTF expression in the MSCs. The mice receiving MSC-CNTF transplantation exhibited significantly improved symptoms with shortened disease course and lessened disease severity. The cell transplantation also resulted in significantly decreased peripheral blood TNF-alpha levels, ameliorated CD3+T cell infiltrations and lowered TNF-alpha expression in the lesions, while the levels of IFN-gamma underwent no significant changes.

CONCLUSIONTransplantation of CNTF gene-transfected MSCs results in decreased peripheral blood TNF-alpha and IFN-gamma levels and reduced inflammatory cells, CD3-positive cells and TNF-alpha expression in the lesion of EAE, therefore providing better effect than MSCs in relieving the symptoms of EAE in mice.

Adenoviridae ; genetics ; metabolism ; Animals ; Bone Marrow Cells ; metabolism ; Ciliary Neurotrophic Factor ; biosynthesis ; genetics ; therapeutic use ; Encephalomyelitis, Autoimmune, Experimental ; therapy ; Female ; Genetic Therapy ; Interferon-gamma ; blood ; Mice ; Mice, Inbred C57BL ; Random Allocation ; Stromal Cells ; metabolism ; T-Lymphocytes ; immunology ; Transfection ; Tumor Necrosis Factor-alpha ; blood

Animals ; Cell Proliferation ; Chemokine CXCL12 ; metabolism ; Hepatocytes ; cytology ; metabolism ; Male ; Rats ; Rats, Wistar ; Receptors, CXCR4 ; metabolism

OBJECTIVETo evaluate the correlation of 64-multidetector-row CT (64MDCT) perfusion imaging with microvessel density(MVD) and vascular endothelial growth factor(VEGF) in colorectal carcinoma.

METHODS64MDCT perfusion imaging was performed in 33 patients with pathologically verified colorectal carcinoma. Time-density curves (TDC) were created from the region of interest (ROI) drawn over the tumor, target artery and vein by 64MDCT with perfusion functional software. The individual perfusion maps generated were for blood flow (BF), blood volume (BV), mean transit time (MTT) and permeability-surface area product (PS). MVD and VEGF expression of surgical specimens were examined by immunohistochemical staining with anti-CD34, anti-VEGF monoclonal antibody. MVD and VEGF were compared among the different types of TDC in colorectal carcinoma. The correlation of CT perfusion parameters with MVD and VEGF was also examined.

RESULTSTDC of colorectal carcinoma was divided into five types according to their shapes. MVD in the colorectal carcinoma was 22.61+/-9.01. VEGF staining was found in 25 of 29 tumors (86.2%). The score of VEGF expression was 4.15+/-1.09. No significant differences of MVD and VEGF expression among TDC types were found (F=2.59, 1.11, P>0.05). There were also no correlations of MVD and VEGF expression with any dynamic CT parameters (P>0.05).

CONCLUSION64MDCT perfusion imaging, MVD and VEGF may reflect angiogenic activity, but no significant correlations are found among them.

Adult ; Aged ; Colorectal Neoplasms ; blood supply ; diagnostic imaging ; Female ; Humans ; Male ; Microvessels ; Middle Aged ; Neovascularization, Pathologic ; Tomography, Spiral Computed ; methods ; Vascular Endothelial Growth Factor A ; metabolism ; Young Adult

OBJECTIVEA prospective study was conducted to probe into the relationship between arterial oxygen partial pressure (PaO2) and brain injury during cardiopulmonary bypass (CPB) in infants with cyanotic congenital heart disease (CHD).

METHODEnrolled in the study were 45 cyanotic infants, who were less than three years old and underwent corrective cardiac surgery from August 1(st), 2010 to January 31(st), 2011 at Guangdong General Hospital. All the infants had a pulse oxygen saturation (SpO2) lower than 85% and were randomly allocated into three groups by a specific computer program. In controlled group 1 (G1 group), PaO2 levels were controlled at 80 - 120 mm Hg (1 mm Hg = 0.133 kPa) during CPB; in controlled group 2 (G2 group), PaO2 levels at 120 - 200 mm Hg during CPB; while in uncontrolled group (G3 group), PaO2 levels were at 200 - 400 mm Hg during CPB. Blood samples were collected just before starting CPB, at the end of CPB, and at 3 h, 5 h, and 24 h after CPB (T1, T2, T3, T4, T5) for the determination of serum concentrations of protein S100β, neuron specific enolase (NSE), and adrenomedullin (ADM) by ELISA.

RESULTProtein S100β rose significantly after starting CPB. In group G3, it reached a peak of (699 ± 139) ng/L by the end of CPB, significantly higher than those in groups G1 and G2 [(528 ± 163) ng/L and (585 ± 155) ng/L], and was positively correlated with PaO2 levels (r = 0.526, P < 0.01). NSE levels of group G1 were continuously rising after starting CPB and reached significantly high levels at 3 h or 5 h after CPB [(12.2 ± 3.4) µg/L and (12.3 ± 3.7) µg/L], while those of group G2 rose significantly during CPB [(10.9 ± 4.8) µg/L] and even higher at 3 h or 5 h after CPB [(12.6 ± 5.1) µg/L and (13.2 ± 5.4) µg/L]. NSE levels of group G3 rose significantly during CPB and maintained at a high level [(12.2 ± 5.7) µg/L] afterwards. There was no significant difference in serum ADM concentrations among different time points in each group and among these three groups. All the infants were discharged from the hospital without any obvious nervous symptom and sign.

CONCLUSIONHigh PaO2 during CPB in infants with CHD might cause an increase of serum protein S100β and NSE, indicating that brain injury might become worse with a higher PaO2 and might be positively correlated with PaO2 during CPB.

Cardiopulmonary Bypass ; Child, Preschool ; Cyanosis ; Female ; Heart Defects, Congenital ; blood ; physiopathology ; surgery ; Humans ; Infant ; Male ; Nerve Growth Factors ; blood ; Oximetry ; Oxygen ; blood ; Partial Pressure ; Phosphopyruvate Hydratase ; blood ; Prospective Studies ; S100 Calcium Binding Protein beta Subunit ; S100 Proteins ; blood ; Serum