Objective To develop a DNA microarray for HLA-DR1,DR51 group genotyping. Methods According to the specific allelic sequences coding HLA-DR1,DR51 loci,HLA- DR1,DR51 group typing probes which were immobilized on a glass support were synthesized.A pair of group-specific primers labeled by the Cy5-dCTP were designed,then the primers were used in the PCR,thus the PCR products were labeled with Cy5.The labeled PCR products were hybridized with array.The signals were scanned by scanner and analyzed by image software.The typing results were confirmed by standard DNA and PCR-SSO. Results A total of 130 samples were typed by this DNA array.There were 34 HLA-DR1,DR51 group loci typed by DNA array.Among them,18 loci were DR15,8 were DR16,6 were DR10 and 2 were DR1.No false positive or false negative typing results occurred.The accuracy and reproducibility were 100% and the overall time of genotyping was about 3.5 hours. Conclusions DNA array technique is a precise,rapid molecular method of high resolution power and high specificity for HLA-DR1,DR51 genotyping,which is applicable to clinical transplant practice.

Objective To develop an oligoneucleotide array for HLA-DR52 group rapid genotyping.Methods According to the special allele sequences of HLA-DRB loci in Chinese Han's population, HLA-DR52 group typing probes which were immobilized on a glass supports were synthesized. A pair of group-special primers labeled by the Cy5-dCTP were designed and were used in the PCR. The labeled PCR products with Cy5 were hybridized with array. The signals were scanned by a scanner and analyzed by Image software. 83 samples were typed by this array and the results were compared with PCR-SSP typing.Results Among 57 HLA-DR52 group loci typed by PCR-SSP,2 samples had no HLA-DR52 loci typed by array,3 DR52 group homozygotes typed by PCR-SSP were actually heterozygotes by array. The other 1 non-DR52 group homozygote identified by PCR-SSP was a heterozygote with one DR52 group locus. Conclusion The oligoneucleotide array technique is a precise, rapid molecular method for HLA-DR52 genotyping. Compared with PCR-SSP method, the genotyping chip is more sensitive and intuitionistic and suitable for clinic practice.

Objective To evaluate the effect of live frog skin in the treatment of deep partial thickness and full thickness burn. Methods 159 burned patients were randomly divided into two groups: wounds treated with frog skin or suckling pig skin. Wounds were covered by frog skin(35 cases) or by suckling pig skin(31 cases) after surgical debridement or when the crust began to dissolve for patients with deep partial thickness burns, and by frog skin (49 cases) or by sucking pig skin(44 cases) after surgical debridment or autologous skin grafting and escharetomy for patients with full thickness burns. There was no significant difference between the two groups in mean age, burn area and burn type. Results Compared with the suckling pig skin, frog skin could remarkably decrease days of wound recovery and antibiotics administration, and reduced incidence of wound infections and rejection in patients with full thickness burns. Conclusions Frog skin possessed more powerful anti-infection ability, and rare rejection phenomenon in the treatment of burn wounds. It may be a suitable biological dressing for burn wounds.

Public Health ; methods ; Systems Analysis

Objective:To study the preventive and health effects of Ampelopsis grossedentata on hyperlipidemia. Method:Thirty Wistar rats were divided into three groups:control group, model group and test group. The control group was fed normal diet, while the model and test groups were fed high fat diet.The control and model group were given water while the test group was given water with 20% Ampelopsis grossedentata for 15 w. Serum lipids, hemorrheological indices, myocardial enzymes and IL-6 were measured. Results: At the end of experiment, serum TC, TG, LDL-C, and apoB/apoA, serum and myocardial MDA, myocardial enzymes activities, hemorrheological indices and serum IL-6 were obviously higher in model group while the activities of serum and myocardial SOD and GSH-Px were lower. In test group. serum TC, TG and apoB/apoA levels, serum and myocardial MDA, myocardial enzymes activities, hemorrheological indices, and serum IL-6 were significantly lower than those in model group, but HDL-C, SOD and GSH-Px activities higher. Conclusion:Ampelopsis grossedentata could significantly decrease blood lipids and hemorrheological indices of rats fed with high fat diet, and protect myocardial cells fromoxidation, and as a result to prevent hyperlipidemia and cardiovascular disease.

OBJECTIVETo study the effect of Flos Daturae alkaloids (FDA) on TGF-beta1-1uuuu;U epithelial-mesenchymal transition (EMT) of human pulmonary adenocarcinoma A549 cells.

METHODSA549 cells in vitro cultured were randomly divided into 5 groups, i.e., the blank control group, the TGF-beta1 group, the low dose FDA group, the medium dose FDA group, and the high dose FDA group. The morphologies of A549 cells were observed. Expressions of cytokeratin (CK)-19 and alpha-smooth muscle actin (alpha-SMA) were detected by Western blot and real-time PCR at 24, 48, and 72 h, respectively.

RESULTSA549 cells in the TGF-beta1, group turned from cobblestone to spindle shape gradually. Those in low, medium and high dose FDA groups showed similar shapes to those of the TGF-beta1 group. There was no statistical difference in the morphology of A549 cells among the 3 dose FDA groups (P > 0.05). Western blot showed that, when compared with the blank control group, the expression of CK-19 was down-regulated, but the expression of alpha-SMA was up-regulated in the TGF-beta1 group (P < 0.01). Compared with the TGF-beta1, group, the expression of CK-19 was up-regulated, but the expression of alpha-SMA was suppressed in low, medium and high dose FDA groups (P < 0.01). The CK-19 expression obviously increased, but the alpha-SMA expression was suppressed in high dose FDA group at 72 h (P < 0.01). Real-time PCR results showed, as compared with the TGF-beta1 group, the mRNA expression of CK-19 was increased, but the mRNA expression of alpha-SMA was reduced in low, medium and high dose FDA groups (P < 0.01).

CONCLUSIONSFDA had no effect on EMT morphological changes of TGF-beta1 induced A549 cells. FDA could reverse characteristic markers of A549 cells during EMT to some extent, such as expressions of CK-19 and alpha-SMA. The expression of CK-19 (as the epithelium marker) increased and the expression of alpha-SMA (as the mesenchymal marker) was reduced. Besides, they were most obviously seen in the high dose FDA group at 72 h in a dose- and time-dependent manner.

Actins ; Adenocarcinoma ; Alkaloids ; pharmacology ; Cell Line, Tumor ; Datura ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; Epithelial-Mesenchymal Transition ; Epithelium ; Humans ; Transforming Growth Factor beta1 ; metabolism

ety, reduce the risk of infection and improve the efficiency of nursing service.

Objective To investigate the computer tomography (CT) and magnetic res-onance imaging (MRI) features of posterior reversible encephalopathy syndrome (PRES) caused by pregnancy-indueed hypertension (PIH). Methods CT scan and MRI scan + diffusion-weighted imaging (DWI) were used in 31 patients with PIH. Results CT scan found low density lesions in parietal-occipital lobe of watershed regions in 31 patients with PIH. CT value was 19 to 23Hu. Among them, MRI showed slight hypointensity lesions on T1-weighted images and slight hyperintensity lesions on T2-weight images in 11 patients, fluid-attenuated inversion recovery (FLAIR) sequence showed hyperintensity lesions, DWI revealed isointense, and apparent diffusion coefficient- did not decrease. The recovery time of clinical symptoms was earlier than that of MRI. Conclusions The lesion sites on cr and MRI had certain characteristics in PIH occurred PRES at late pregnancy, combining clinical history a clear diagnosis could be made.

Objective To investigate correlation between Helicobacter pylori infection and incidence of myocardial infarction and cerebral infarction ,and serum homocysteine(Hcy) level .Methods A total of 72 patients with myocardial infarction ,86 patients with cerebral infarction and 80 healthy subjects were enrolled and detected for Hp‐IgG and serum Hcy by using enzyme‐linked im‐munosorbent assays and enzymatic cycling method .Relationship between Hp infection and serum level of Hcy were analyzed .Results The positive rate of Hp‐IgG and serum Hcy level in patients with myocardial infarction were significantly higher than healthy subjects(P<0 .05) .Among patients with myocardial infarction ,serum level of Hcy in Hp‐IgG positive patients was higher than that in Hp‐IgG negative patients(P<0 .05) .The positive rate of Hp‐IgG between patients with cerebral infarction and healthy subjects was without significant difference(P>0 .05) .Among patients with cerebral infarction ,serum level of Hcy between Hp‐IgG positive patients and Hp‐IgG negative patients were without significant difference(P>0 .05) .But serum Hcy level in patients with cerebral infarction was significantly higher than that in healthy subjects(P<0 .05) .Conclusion Hp infection might promote the occurrence of myocardial infarction by affecting serum level of Hcy .However ,there might be without obvious correlation between Hp infection and the occurrence of cerebral infarction .And there could be no direct association between high serum level of Hcy in patients with cerebral infarction and Hp infection .