Objective To develop an ELISA for detection of anti-dsDNA antibody by using plasmid DNA as antigen.Methods DNA in plasmid pBV220 for prokaryotic expression vector was purified by base-cleavage. The microtiterplates were pretreated by poly-L-lysine and coated by the plasmid DNA in a dilution of 1∶50 as antigen. An ELISA method for detection of serum anti-dsDNA antibodies was developed with HRP-SPA as enzyme-labeled marker.The IIF using crithidia lucilia as substrate was performed simultaneously for comparison. The serum samples from 64 patients with SLE, 8 with MCTD and 17 with RA were detected. Results The concentration of DNA was 1 54 g/L by UV spectrophotometer at wavelength of 260 nm. The positive percentage of ELISA for anti-dsDNA was higher than that of IIF. By comparison with IIF the positive percentages in SLE, MCTD and RA groups were 23 4% vs 17 2%, 12 5% vs 12 5% and 11 8% vs 5 9%, respectively, and the coincident rates between the 2 methods were 93 8%, 100% and 94 1% respectively. The sensitivity and specificity of the developed ELISA for detection of anti-dsDNA were 100% and 93 4% when IIF was as gold standard.Conclusion The ELISA by using plasmid DNA as antigen to detect anti-dsDNA has fine precision, sensitivity and specificity. Its positive rate is higher than that of IIF thus it will contribute to monitor the activities for SLE patients′ condition.

OBJECTIVE: To investigate the audiologic characteristics of the children with cleft lip and/or palate. METHOD: Sixty-two children with cleft lip/palate were enrolled in the study. Tympanometry, DPOAE, ABR were tested in all the chidren. RESULT: Regarding the ABR threshold as the diagnostic criteria, 51 (41.13%) ears had hearing loss, mainly moderate. Sixty-three (59.43%) ears had abnormal tympanometry, which meant the disorder function of the middle ear. Twenty-six (56.52%) ears didn't pass the DPOAE. CONCLUSION The proportion of the hearingloss in the children with cleft lip/palate was large. There was a good coherence among the three objective examinations when assess the children with cleft lip/palate.
Audiometry ; Child ; Child, Preschool ; Cleft Lip ; physiopathology ; surgery ; Cleft Palate ; physiopathology ; surgery ; Female ; Humans ; Infant ; Male ; Postoperative Period

Objective: To learn anxiety, depression, sleep quality and quality of life among patients with primary pruritus ani, and to analyze their correlation with anal itching symptom, so as to provide insights into the prevention and treatment of primary pruritus ani. Methods: Patients with primary pruritus ani were selected from anorectal department of the First Affiliated Hospital of Zhejiang Chinese Medical University. The Visual Analogue Scale, pruritus frequency score, Hospital Anxiety and Depression Scale, Pittsburgh Sleep Quality Index and Dermatology Life Quality Index (Chinese version) were used in the survey. Spearman rank correlation analysis was employed to analyze the correlation between the degree and frequency of pruritus ani with, anxiety, depression, sleep quality and quality of life. Results: Sixty patients with primary pruritus ani were investigated, including of 7 males and 53 females, with an average age of (34.37±10.30) years. The patients with mild, moderate and severe pruritus accounted for 28.33%, 48.33% and 23.34%, respectively. The patients with pruritus frequency of grade 1, 2 and 3 accounted for 51.67%, 38.33% and 10.00%, respectively. The patients with anxiety, depression, and coexistence of anxiety and depression accounted for 18.33%, 13.33% and 10.00%, respectively. The patients with very good, good, average and poor sleep quality accounted for 15.00%, 46.66%, 30.00% and 8.34%, respectively. No, mild, moderate and severe impacts on quality of life among the patients accounted for 10.00%, 46.66%, 33.34% and 10.00%, respectively. The degree of pruritus was positively correlated with anxiety (rs=0.405, P=0.001), depression (rs=0.343, P=0.007), sleep quality (rs=0.293, P=0.037) and quality of life (rs=0.338, P=0.008). However, there was no significant correlation between the frequency of pruritus with the above factors (all P>0.05). Conclusion The degree of pruritus in patients with primary pruritus ani is related to anxiety, depression, sleep quality and quality of life.

Objective To explore the clinical features, imaging and their diagnosis vaule of Baló's concentric sclerosis (BCS). Methods The clinical manifestation, laboratory examination, neuroimaging characteristics, pathology and follow-up of 12 (2 outpatients and 10 inpatients) cases of BCS were analyzed retrospectively. Results The onset age of 12 patients (male 4, female 8) ranged from 8 to 54 years old (mean 36.8 years). Unconcerned and slowness response were common in the disease onset (7 cases).Other symptoms and signs such as hypophrenia ( 10 cases), limitation of limb or hand movement (8 cases),urinary and fecal incontinence (7 cases), hypo-myodynamia (7 cases), positive pyramidal sign (6 cases)were not rare in course of disease. Eight cases underwent the examination of cerebrospinal fluid myelin basic protein, which increased in 7 cases. Demyelinating change and perivascular inflammatory infiltration were shown in 6 brain biopsies. The involvements of multi-lesions were found in 11 cases. Four patients underwent CT scanning and all the cases showed with hypodense lesions and couldn' t be enhanced. On MRI, all cases appeared round or round-like in shape. Enhanced lesions showed ring-like or half-ting-like shape (8 cases) and concentric ring (3 cases). It's easy to be misdiagnosed as brain tumor (8 cases).Conclusions BCS is a kind of demyelinating disease of subacute or chronic onset accompanied with cognitive disorder being the most common initiating symptoms. It's prone to be misdiagnosed. Diagnosis depends on MRI and corticosteroid treatment is effective. BCS has a good prognosis and presents with benign clinical course.

Objective To study effect of piracetam combine with hyperbaric oxygen in treatment of acute carbon monoxide poisoning patients with electrocardiogram and its effects on Lactate clearance.Methods 60 patients of acute carbon monoxide poisoning who received therapy from February 2011 to February 2016 in our hospital were selected as research objects.All accord with the diagnostic criteria of acute carbon monoxide poisoning.According to draw method,those patients were divided into the experimental group(n=30)and the control group(n=30).The two groups were given a large number of sustained oxygen,intracranial pressure,protect brain cells,promote blood circulation and improve microcirculation and other basic symptomatic treatment.The control group on the basis,was treated with hyperbaric oxygen,one times a day,a total of ten times.while the experimental group was treated with piracetam combine with hyperbaric oxygen,hyperbaric oxygen method with the control group,intravenous drip of Piracetam and Sodium Chloride Injection,each 100ml,two times a day,a total of treatment for ten days.Then abnormal ECG,creatine kinase isoenzymes(CK-MB),troponin(cTnl),lactate clearance,incidence of delayed encephalopathy,mortality,therapeutic effect of two groups were compared.Results ECG abnormal rate there was no difference between the two groups before treatment,after treatment,the abnormal rate of the experimental group was significantly lower than the control group [6.66(2/30)vs.33.33％(10/30)](P<0.05); CK-MB、cTnl、6h and 24h after treatment,Lactate clearance rate was significantly higher than control group[(15.80±2.03)％vs.(10.26±2.01)％,(20.75±3.12)％vs.(13.07±2.56)％](P<0.05);DEACMP rate and mortality was significantly lower than the control group[6.66％(2/30)vs.33.33％(10/30),3.33％(1/30)vs.30.00％(9/30)](P<0.05); The total effective rate was significantly higher than the control group[95.56％(28/30)vs.75.56％(22/30)](P<0.05).Conclusion Piracetam combine with hyperbaric oxygen is well for acute carbon monoxide poisoning,which can improve the clearance rate of lactic acid,improve hypoxia and myocardial injury,and reduce the abnormal ecg.

Objective To construct five shRNA-expression plasmids and to investigate the expression of Smad2 in TGF-?/ Smads signal transduction treated with shRNA-expression plasmid.Methods Five shRNA-Smad2 DNA sequences from mRNA sequence of mouse Smad2 gene were designed and synthesized.DNA oligonucleotides encoding an appropriate shRNA were inserted to shRNA expression vector respectively.Five shRNA-Smad2 expression plasmids were obtained and then transfected into NIH/3T3 cells.The suppressed expression of Smad2 was assessed by RT-PCR and Western-blotting.Results The shRNA-expression plasmid numbered 2.4 could markedly reduce the expression of Smad2.The suppression effect of the RNAi-pool composed of four different plasmids was more obvious than that of any single.Conclusion The shRNA-expression plasmids were successfully constructed,which could specifically and effectively suppress the expression of Smad2.The method of using a mixture of RNAi plasmids to improve the RNAi efficiency was established.

OBJECTIVETo compare the effect of recombinant OPG-Fc and recombinant RANK protein on the differentiation of osteoclast precursors.

METHODSMouse osteoblasts cell lines were incubated with osteoclast precursors cell lines RAW 264.7 for 9 days with 10(-5) g/L rhRANK or rhOPG-Fc or PBS added to the coculture system. TRAP stain positive cells counting and cortical bone pit formation counting were performed in the 9th day.

RESULTSMultinuleated TRAP stain positive cells were observed in the cocluture systems after 6 days incubation,and plenty of mature osteoclasts could be observed in the 9th day. With the addition of 10(-5) g/L rhOPG-Fc or rhRANK, multinucleated giant cells and cortical bone pit formation couting decreased significantly compared with the control group, and the rhRANK group decreased more significantly than the rhOPG-Fc group.

CONCLUSIONSBoth rhOPG-Fc and rhRANK can inhabit the differentiation of osteoclast precursors and prevent them forming mature osteoclasts,moreover,the rhRANK shows the significant inhabition effect than the rhOPG-Fc.

Animals ; Cell Differentiation ; drug effects ; Cells, Cultured ; Mice ; Osteoclasts ; cytology ; drug effects ; Osteoprotegerin ; pharmacology ; Receptor Activator of Nuclear Factor-kappa B ; pharmacology ; Recombinant Proteins ; pharmacology ; Stem Cells ; cytology ; drug effects

ObjectiveTo introduce a new technique to create a pulmonary valve biorifice for reconstruction of right ventricular outflow tract in tetralogy of Fallot (TOF),and to summarize its initial clinical experience and therapeutic effect.MethodsThe new technique regarding reconstruction of right ventricular outflow tract with a pulmonary valve biorifice was used in a total of 53 TOF cases (the observation group).The conventional technique regarding reconstruction of right ventricular outflow tract was used in other 50 TOF cases (the control group).The clinical dates of all cases were reviewed retrospectively.ResultsThe ages,weights,cardiopulmonary bypass time,cardiac arrest time,as well as the post operation ventilation support time were not different significantly between two groups.Compared with the contrul group,patients from the observation group had shorter duration of ICU stay.After operation,in the observation group,only 2 cases had large amount of pleural effusion,1 case meddle,and 8 cases little amount of pleural effusion; whereas,in the control group,the corresponding numbers were 1,5 and 17,respectively.At the time point of 1 week after operation,all patients were rechecked by echocardiography,no pulmonary valve stenosis was found.Moderate pulmonary valve regurgitation was found in 8 cases,mild regurgitation in 15 cases from the observation group; and severe regurgitation in 3 cases,moderate regurgitation in 17 cases,and mild regurgitation in 16 cases from the control group.A total of 33 cases from the observation group were rechecked at the time point of half year after operation,and moderate - mild pulmonary regurgitation were found in 3 cases.A total of 18 cases of them were rechecked 1 - year latter,no pulmonary regurgitation was found.ConclusionsThe new technique to create pulmonary valve biorifice can reduce the pulmonary valve regurgitation and postoperative pleural effusion,and improve the early outcomc.

The formation of osteolytic bone lesions is a key process for osteolytic cancer to metastasize to the bone and is under the control of a set of transcription factors. Recently, the inhibitor of differentiation 1 (Id1) has been linked with angiogenesis, tumorigenesis, metastasis and bone formation. However, the function of Id1 during the process of bone destruction caused by cancer in vivo has not yet been elucidated. We, therefore, examined whether and how Id1 affects the ability of cancer to form osteolytic lesion in vivo. The study used a lentiviral vector overexpressing short hairpin RNA (shRNA) targeting Id1 gene. PC3 cells, a prostate cancer cell line, were transduced with Id1 shRNA or negative control (NC) shRNA before implantation in BALB/c mice. Cells were implanted in a tibial injection model. Tumor formation in bone was monitored by X-ray. The relationship between parathyroid hormone-related protein (PTHrP), an osteolytic factor, and Id1 was analyzed by using immunohistochemistry in tissue sections from osteolytic lesion of the BALB/c mice. Our results showed that Id1 shRNA delivery to PC3 cells by lentivirus caused efficient and stable Id1 gene silencing. In the intratibial model, PC3 cells produced primarily osteolytic lesions in the bone. Eleven of 14 mice in Id1 shRNA group but only 4 of 14 mice in the NC shRNA group developed osteolytic lesions with cortical destruction at 4th week. Mice treated with Id1 shRNA had larger tumor volume in the bone and larger cortical destruction. The expression of PTHrP protein in PC3 cells was not affected by Id1 knockdown in vivo. These results indicate that Id1 may down-regulate the ability of PC3 cells to form osteolytic lesions in vivo and the signal pathway needs to be further investigated.

0.05).Conclusion The result suggests that T1 locus genetic polymorphism is weakly associated with asthma.