Objective To purify and quantify the autoantibodies against various epitopes within BP180-NC16A domain from the sera of patients with bullous pemphigoid (BP). Methods Three epitopes within BP180-NC16A domain, NC16A-1, NC16A-2 and NC16A-3, were prepared. Blood samples were obtained from 10 patients who were diagnosed as active BP by clinical, pathological and immunofluorescence examination. Autoantibodies against these epitopes were purified with affinity chromatography column from the sera of these patients with active BP, and quantified separately. The relative binding affinity of autoantibodies to NC 16A-1, NC 16A-2 and NC 16A-3 was measured using thiocyanate elution method. Results The autoantibodies against NC16A-1, NC16A-2 and NC16A-3 were successfully purified from the sera of patients. On average, the amount was 49.0±20.7 μg, 117.7±22.4 μg and 39.5±18.9 μg respectively for autoantibodies to NCI6A-1, NC16A-2 and NC16A-3 purified from a portion of serum containing about 20 mg IgG. Both the amount and binding affinity of anti-NC16A-2 autoantibody were significantly higher than those of anti-NC16A-1 and anti-NC16A-3 autoantibodies. Conclusion BP180 NC16A-2 (aa507-aa520) may be the major epitope recognized by pathogenic autoantibodies in patients with BP.

Objective To evaluate the application value of serum endotoxin combined with integrated model for end-stage liver disease (iMELD) in evaluating the short and medium term prognosis of patients with decompensated liver cirrhosis.Methods The clinical data of 68 patients with decompensated liver cirrhosis were retrospectively analyzed.The patients were divided into survival group and death group according to the result of follow-up at the 3,6 and 12 months.The iMELD score and Child-Turcotte-Pugh (CTP) score were calculated according to relevant indicators,and the serum endotoxin level was detected.The correlation between serum endotoxin and iMELD score was analyzed in the short and medium term prognosis in patients with decompensated liver cirrhosis.Results After 3 months' follow-up,5 patients died.The iMELD scores in survival group and death group were (39.26 ± 7.29) and (49.29 ± 8.63) scores,CTP scores were (9.32 ± 2.12) and (12.03 ± 2.75) scores,serum endotoxin levels were (9.21 ± 2.24) and (15.39 ± 5.12) ng/L,and there were statistical differences (P ＜ 0.05).After 6 months'follow-up,12 patients died.The iMELD scores in survival group and death group were (41.35 ± 8.03) and (52.18 ± 10.37) scores,CTP scores were (9.78 ± 2.05) and (12.93 ± 1.99) scores,serum endotoxin levels were (8.96 ± 2.41) and (16.43 ±6.03) ng/L,and there were statistical differences (P ＜0.05).After 12 months' follow-up,19 patients died.The iMELD scores in survival group and death group were (40.74 ± 6.38) and (52.29 ± 8.53) scores,CTP scores were (10.01 ± 2.23) and (13.27 ± 1.69) scores,serum endotoxin levels were (8.53 ± 2.34) and (16.52 ± 6.08) ng/L,and there were statistical differences (P ＜ 0.05).The area under curve of receiver operating characteristic (ROC) was 0.952,standard error was 0.048,and 95％ confidence interval was 0.790-1.022.The best critical value of iMELD score to predict death was ≥48.00 scores,and the risk of death would rise if iMELD score was increased.The best critical value of serum endotoxin to predict death was ≥ 11.00 ng/L.Conclusion iMELD score and serum endotoxin have important value in predicting prognosis in patients with decompensated liver cirrhosis.

Objective To establish a usable method for the amplification of human diversified immunoglobulin genes.Methods Peripheral blood mononuclear cells (PBMCs) were isolated from healthy individuals and in which RNA was extracted and reverse-transcripted into cDNA.? light chain genes and Fd fragments of human IgG and IgM were amplified by PCR with designed primers and identified by gel analysis and DNA fingerprinting.Results All the intent immunoglobulin genes were successfully amplified and the obtained products were identified correct and with very good diversity.Conclusion It is feasible to amplify human diversified immunoglobulin genes from PBMCs by RT-PCR using properly designed primers.This method will be useful in the studies of human antibodies,correlative immunological molecules and autoimmune diseases.

Objective:To explore effectiveness of Russell Barkley's parent training in children with comorbid attention deficit hyperactivity disorder(ADHD) and oppositional defiant disorder(ODD),and to evaluate its applicability in mainland China.Methods:Parents of 30 DSM-Ⅳ ADHD+ODD children were given 10 weekly trainings.Results:Total numbers of symptoms,symptoms of ADHD,symptoms of inattention,hyperactivity/impulsivity and ODD declined significantly(P

Objective:To explore risk factors of attention deficit hyperactivity disorder(ADHD) comorbid with oppositional defiant disorder(ODD) in children.Methods:Parents of 43 pure ADHD+ODD children,and 43 pure ADHD children(criteria of DSM-Ⅳ;group matched by sex,age and ADHD subtypes) were asked to complete a set of investigation questionnaires including child and family information;family history;pregnancy,delivery and infancy complications;physical development and temperament;and predisposing factors and parents' or teachers' responses toward children's misbehaviors.Results:Probabilities of psychosis or schizophrenia of family members,long-term conflict between parents and parents' constant/stringent criticism to children in ADHD+ODD group were significantly higher than those in ADHD group(psychosis or schizophrenia of family members(16% vs 2%,P=0.058,OR=8.167);long-term conflict between parents(23% vs 7%,P=0.035,OR=4.040);parents' constant/stringent criticism to children(51% vs 28%,P=0.027,OR=2.706);probability of having anoxia at birth in ADHD group was significantly higher than that in ADHD+ODD group(12% vs 0,P=0.055,OR=0.469).Setting diagnosis of ADHD+ODD as dependent variable,and factors with significant difference between the two groups as independent variables,after 2 steps of regression analyses,one factor,i.e.parents' constant/stringent criticism to children was left in the model(P=0.054,OR=2.879).The regression formula had a sensitivity of 53.5%,a specificity of 83.7%,and total correction rate of 68.6%.Conclusion:Inappropriate parenting is a risk factor of ADHD comorbid with ODD,and long-term conflict between parents and positive family history of psychoses might have influence on ADHD comorbid with ODD.

Erythrocytes were frozen with DMSO of different concentrations (not over 20%) as a cryoprotec-tive agent in freezers of different temperatures.The rate of temperature drop was not controlled. Of the erythrocytes actually frozen, the average recovery rate after thawing was 91.3 per cent (range 87.5~ 95%). The average survival rate of erythrocytes stored at -20℃, -30℃, or -80℃ for up to eight months was 90.9 per cent (range 88.2~ 94.5). As compared with glycerol, which is in common use internationally, DMSO seems to be a better cryoprotective agent.

Objective To study the CT and high resolution CT (HRCT) features of peripheral pulmonary hamartoma, and improve the diagnostic accuracy. Methods CT and HRCT materials of fifteen peripheral pulmonary hamartomas with pathologically proved were analyzed retrospectively. Results In the 15 cases, 6 and 9 were located in right and left lung respectively. All of the 15 cases were solitary lesion. The sizes of the lesions were from 0.9 cm?1.0 cm to 3.8 cm?4.5 cm. Most of the lesions were under the pleura. The shapes of the lesions were round and oval. The margins of the lesions were clear. The average CT density was (39.6?13.2) HU,and fat was shown in 6 cases(40%), calcification was shown in 8 cases(53.3%).Conclusion Peripheral pulmonary hamartoma with specific CT and HRCT features can be diagnosed correctly.

The unicellular red alga, Porphyridium , can be artificially cultivated. Some of very useful bioactive substances such as phycobiliproteins? polyunsaturated fatty acids (PUFA) and sulfated polysaccharides, which are synthesize by Porphyridium , have promising applications. This paper deals with the study of the character, production and application of Porphyridium.

BACKGROUND: There are many advantages in treatment of deglovinginjury in hand with compatible grafting of temporoparietal fascial flaps(TFFs), while there are few relevant reports in China.OBJECTIVE: To evaluate function and appearance of hands after restora-tion of soft tissue defects with compatible grafting of TFFs.DESIGN: Case retrospective analysis.SETTING: Department of Maxillofacial Surgery, Plastic Surgery Hospital,Peking Union Medical College, Chinese Academy of Medical Sciences.PARTICIPANTS: Eleven patients with hand injuries received compatiblegrafting of TFFs in the Department of Maxillofacial Surgery of PlasticSurgery Hospital, Peking Union Medical College, Chinese Academy ofMedical Sciences from January 2001 to May 2005 were selected including8 males and 3 females with a mean age of 30 years. All subjects participat-ed in the observation and analysis voluntarily. All patients presented expo-sures of tendon, bone or joint.METHODS: Preoperative location was done with Doppler or palpation methods and the orientation of temporal vascular nerve tracts were labelled. Re-cutization of surface of fascial flap was achieved through fullthickness skin transplantation. Sizes of fascial flaps were limited between 10 cm×9 cm and 4 cm×6 cm. Follow-up was 8 months (6-11 months),the items of which included postoperative functional inspection and satisfactory degree of appearance. Ratio of total active range of motion (AROM) with joints injured and corresponding uninjured joints were used for functional evaluation, and the meso was zero. Functional rehabilitation was divided into 3 grades according to the ratio: Good as the ratio above 0.75, fair as the ratio between 0.5 and 0.75, poor as the ratio below 0.5. Evaluation on appearances of hands contained satifactions of both patients and doctor including whether there were fat and clumsy or visible scars.MAIN OUTCOME MEASURES: Postoperative functional recovery of hand after compatible grafting of TFFs.RESULTS: Eleven patients with hand injury were included and 4 patients withdrew from the study. ①Fascial flaps of 9 patients completely survived,one patient got partial necrosis due to venous injury during flap harvesting,and wounds spontaneously healed by exchanging dressing. Venous crisis was found in the 3rd day of postoperation in one patient during operation research (OR) and there were thromboses in anastomosed veins due to the folding of overlong vein, which was unobstructed after reanastomosis, while there were sporadical scars in the raw surfaces. ②Functional recovery of hands were good in 4 cases and better in 3 cases, 4 cases withdrew from the study. Both doctors and patients were satisfied with the recovery status of appearance of hands.CONCLUSION: It is better to simultaneously adapt compatible grafting of TFFs and re-cutization in restoration of tissue defects in hands such as exposures of bone and tendon. Better aesthetic results can be achieved with out second flap restoration. Most of all, the supple texture of fascial tissues provides a good gliding environment for functional recovery of tendon and joint, moreover, scars in donor sites are well camouflaged without functional lesions.

Objective To investigate the expression of Ptch-1 and Gli-1, hedgehog pathway-related genes in squamous cell carcinoma (SCC), and the effect of cyclopamine, a specific inhibitor of hedgehog signaling pathway, on the proliferation of a SCC cell line Tca. Methods Skin samples were resected from 42 patients with SCC and 10 normal human controls. Immunohistochemistry and in situ hybridization were employed to study the expression and distribution of Ptch-1 and Gli-1 in these specimens. Tca cells were incubated with cyclopamine (1, 2, 5, 10 μmol/mL) for 48 hours, or cyclopamine (5 μmol/mL) for 1-8 days. The same concentrations of lycopersicin served as the control treatment. Then, MTT assay was performed to detect the proliferation of Tca cells. A fraction of Tca cells were cultured in the presence of 5 μmol/mL cyclopamine for 72 hours followed by BrdU assay for the evaluation of cell growth and proliferation. Results A significant increment was shown in the expression of both Patch-1 and Gli-1 by immunohistochemistry (χ2= 5.656, 6.732, P<0.05, 0.01, respectively) and in situ hybridization (χ2=6.787, 9.600, respectively, both P<0.01) in SCC tissue compared with the control specimens. And both of them were predominantly distributed in the cytoplasm of SCC cells. As MTT assay revealed, cyclopamine notably inhibited the proliferation of Tea cells, and the effect increased with the concentration and action time of cyclopamine. Further more, the percentage of BrdU-positive cells was 26% in cyclopamine-treated Tca cells, significantly higher than that in the blank control cells (77%) and lycopersicin-treated cellls (72%). Conclusions Hedgehog signaling pathway is activated in the lesions of SCC, and inhibition of the pathway may facilitate the treatment of SCC.