Objective To investigate the characteristics and clinical application value of anti-double stranded DNA antibody de-tected by Crithidia indirect immunofluorescence assay method and enzyme linked immunosorbent assay method.Methods Eighty-five patients with systemic lupus erythematosus,20 disease controls and 75 healthy controls were selected.The serum anti-double stranded DNA antibody was detected simultaneously by the methods of Crithidia indirect immunofluorescence assay and enzyme linked immunosorbent assay and their diagnostic efficacies for detection were compared.Results For each method the positive rate in the systemic lupus erythematosus group was significantly higher than that in the disease control group and healthy control group. The difference had statistical significance (P <0.05).The positive rates of Crithidia indirect immunofluorescence assay and enzyme linked immunosorbent assay in the systemic lupus erythematosus group were 72.94% and 88.24% respectively,and the positive predictive value of enzyme linked immunosorbent assay is lower(P <0.05).Meanwhile the anti-double stranded DNA antibody con-centrations detected by enzyme linked immunosorbent assay method showed statistically significant difference among the active sys-temic lupus erythematosus group,the stable systemic lupus erythematosus group and the control group (P <0.05 )and presented linear trend.Conclusion Using Crithidia indirect immunofluorescence assay method to detect anti-double stranded DNA antibody for the systemic lupus erythematosus group has high specificity and is helpful for the diagnosis of systemic lupus erythematosus. Enzyme linked immunosorbent assay can be used to detect anti-double stranded DNA antibody concentration quantitatively,which is linearly related with systemic lupus erythematosus activity and the method is of high sensitivity,which can effectively screen the pa-tients with systemic lupus erythematosus.

Aim To study on the diuretic effect of a phenylphthalazines compound PU1424 and its influence on electrolyte balance, glucose and lipid metabolism, hepatic and renal functions.Methods Male Sprague Dawley rats were randomly divided into solvent control group,PU1424 treated group and HCTZ treated group.Urine was collected per 6 h and blood samples were collected at the end of drug administration.Urinary osmolality was measured by Freezing Point Osmometer;urea concentration was measured by Urea Detection Kit;ion level, blood glucose level, blood lipid level, hepatic and renal function were analyzed by Automatic Biochemical Analyzer.Results Compared to the solvent control group, and urine output of rats treated with PU1424 and HCTZ was increased as 1.52 times and 1.78 times and water intake increased as 1.42 times and 1.56 times respectively.Urine osmolalities were decreased as 61.5% and 50.4% of the control group, and urine urea concentration was decreased as 57.1% and 56.8% of the control group.Urinary electrolytes were decreased by administration of PU1424 and HCTZ compared to the intact plasma electrolytes.The blood glucose levels and blood lipid levels of rats treated with PU1424 had no changes, while the blood glucose and total cholesterol were increased by administration of HCTZ.The urea nitrogen, creatinine, alkaline phosphatase and total protein were intact by administration of PU1424 and HCTZ except the alanine/straw ration increased by HCTZ.Conclusion New diuretic candidate compound PU1424 displays significant diuretic effect with electrolyte balance, blood glucose level, blood lipid level, hepatic and renal function intact.

OBJECTIVETo study the impact of gender factor on the candidate gene study of essential hypertension (EH).

METHODSThe polymerase chain reaction (PCR) was performed to analyze the ACE gene I/D polymorphism of hypertensive patients (50 men and 50 women) and normal controls (50 men and 50 women). The investigation was further focused on possible influence of sex proportion on the conclusion of this kind of research.

RESULTSThe frequency of DD genotype in male hypertensive patients is significantly higher than that in male normal controls (chi(2) = 6.98, P = 0.004). The frequency of D allele in male EH group is significantly higher than that of male normal controls (chi(2) = 6.87, P = 0.009), while no significant difference was observed for II and ID genotype between male EH group and control group (P > 0.05). For female EH group and normal controls, there were no significant differences in frequency of genotype and allele (P > 0.05), the distribution ratio of DD genotype in male EH group is significantly different from that of female EH group (chi(2) = 4.06, P = 0.044). Furthermore, males with DD genotype in EH group had higher SBP and PP than that of males with II and ID genotype (P < 0.05). However, there was no significant difference in DBP in all three genotypes (P > 0.05). At the same time, there was no difference in SBP, DBP and PP (P > 0.05) between II and ID genotype in male EH group. In female hypertensive patients, there was no significant difference in SBP, DBP and PP between all three genotypes (P > 0.05).

CONCLUSIONSThe relationship between DD genotype in male and EH (especially SBP and PP) is closer than any other genotype-EH relationships in both male and female. The gender factor, as a probable confounding factor, can affect many candidate gene studies of essential hypertension including ACE gene I/D polymorphism, and thus biases the conclusion.

Aged ; Female ; Gene Frequency ; Genotype ; Humans ; Hypertension ; genetics ; Male ; Middle Aged ; Peptidyl-Dipeptidase A ; genetics ; Polymorphism, Genetic ; Sex Factors

OBJECTIVEAs newly, functional, diagnostic measure-18F-deoxyglucose single photo emission computed tomography (18F-FDG SPECT-CT) has well sensitivity and specialty , it was proved availability in differentiating benign and malignant tumor. In this study, it was compared the efficiency of detecting residual tumor of thyroid cancer between 18F-FDG SPECT-CT and CT-TWIN.

METHODSDuring May 2004 to August 2004, 56 patients with thyroid disease were divided into two groups, one group was 32 cases suffered thyroid tumor, another was 24 cases suffered improper operation. All cases must take 18F-FDG SPECT-CT and CT-TWIN.

RESULTSForty eight cases had been performed operation, and 8 cases waited and watched because they didn't be found residual tumor by 18F-FDG SPECT-CT and CT-TWIN. The results in detecting thyroid tumor showed that the same rate of diagnostic correction of SPECT-CT was 87.5% (28/32), and the Youden index of SPECT-CT was 0.667, higher than that of CT which was 87.5% (28/32) and 0.633 respectively. When the results two methods were accordant, the rate of diagnostic correction was 100% (24/24), and Youden index was 1, and the result had statistic signification. The rate of diagnostic correction of SPECT-CT in detecting residual tumor was 84.6%, and its Youden index was 0.675. The rate of diagnostic correction of CT in detecting residual tumor was 75.0%, and its Youden index was 0.492; and the result had statistics signification by being compared two different measures.

CONCLUSIONSSPECT-CT and CT all had higher diagnostic efficacy, and they could promote the diagnostic efficacy when they have the same diagnosis in detecting thyroid tumor. The rate of detecting residual tumor of SPECT-CT was 81.3%, compared with that of CT the diagnostic efficacy has greater raise.

Adolescent ; Adult ; Aged ; Child ; Female ; Fluorodeoxyglucose F18 ; Humans ; Male ; Middle Aged ; Neoplasm, Residual ; diagnostic imaging ; Thyroid Neoplasms ; diagnostic imaging ; pathology ; Tomography, Emission-Computed, Single-Photon ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVETo ascertain whether connexin 26 (Cx26) gene was a nuclear modifier gene in an extensive family with matrilineal nonsyndromic deafness associated with A1555G mutation in Huaiyin, China.

METHODSFollowing PCR-restriction fragment length polymorphism (PCR-RFLP) with ApaI restriction enzyme, Cx26 genes from 26 cases, with A1555G mitochondrial mutations in this family, and 62 controls (including 2 patrilineal relatives, 10 spouse controls and 50 unrelated controls), were sequenced.

RESULTSCompared with the reference sequence of Cx26 gene, totally four kinds of nucleotide changes,79G -->A, 109G-->A, 341G-->A and 235delC, were detected in a heterozygous form. However, the former three were previously reported polymorphisms, and only the 235delC was a previously described recessive mutation associated with most autosomal nonsyndromic sensorineural hearing loss in Japan and China. Further study showed that the heterozygous 235delC mutation existed in both one individual with mild hearing loss and two individuals with normal hearing. Clinical characterization showed that 235delC mutation did not seem to modify the deafness phenotype due to the A1555G mutation. Moreover, this 235delC mutation was deduced to derive from a married-in control. Finally, there were no co-segregation between the phenotypes of hearing loss and the genotypes for Cx26 genes based on the four kinds of nucleotide changes.

CONCLUSIONSThe heterozygous 235delC mutation of the Cx26 gene may not modulate the severity of hearing loss associated with A1555G mutation and Cx26 gene is unlikely to be a modifier gene for hearing loss due to A1555G mitochondrial mutation in this Chinese family.

Adolescent ; Adult ; Case-Control Studies ; Child ; Child, Preschool ; China ; epidemiology ; Connexin 26 ; Connexins ; genetics ; Deafness ; epidemiology ; ethnology ; genetics ; Female ; Genotype ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Mutation ; Pedigree ; Phenotype ; Polymorphism, Restriction Fragment Length ; Sequence Analysis ; Young Adult

BACKGROUNDPeptide nucleic acid (PNA) has many characteristics useful in molecular biology. This paper described an effective way to raise the cell ingestion rate of PNA so as to kill gastric cancer cells.

METHODSHeteroduplexes of PNAs and oligonucleotides, wrapped by Lipofectamine 2000, were used to infect SGC7901 cells. The inhibitive effect of heteroduplexes was evaluated by analyzing cell clone forming and cell growth rate. Telomerase activity of SGC7901 cells was detected by polymerase chain reaction enzyme-linked immunosorbent assay (PCR-ELISA) and silver staining assay.

RESULTSPNAs showed a dose-dependent inhibition of cell proliferation. The percentage of proliferation inhibition was 99.4% after 7 days; the rate of cloning inhibition was 98.2% after 8 days; whereas for oligonucleotide groups, at the same concentration, the percentages were 50.1% and 67.5% respectively. Antisense PNA-DNA-Lipofectamine 2000 group (AP-D-L group) exhibited significantly different percentages from the control groups (P < 0.05). The test result indicated that telomerase activity of the AP-D-L group was inhibited (P < 0.05). At the same time, the impact on cell morphology was observed.

CONCLUSIONSThe results showed that PNAs are potent antisense reagents. The telomerase-associated therapies are very promising for the treatment of malignant tumours.

Cell Division ; drug effects ; Cell Line, Tumor ; DNA-Binding Proteins ; Humans ; Peptide Nucleic Acids ; therapeutic use ; Stomach Neoplasms ; pathology ; therapy ; Telomerase ; antagonists & inhibitors ; metabolism ; Transfection

OBJECTIVERecent study demonstrated that hypoxia could regulate the proliferation and differentiation of neural stem cells in vitro. In the present study, effects of low glucose and/or hypoxia on the proliferation and metabolism of neural stem cells were investigated in vitro.

METHODSThe neural stem cells were isolated from the rat embryonic mesencephalon (E13.5), and exposed to different oxygen concentrations (low oxygen: 3% O2 or normoxia: 20% O2) and different glucose concentrations (high glucose concentration: 4.5 g/L and low glucose concentration: 1.4 g/L) for 3 days. The proliferation of neural stem cells were examined by CCK-8 assay. Furthermore, the content of glucose, lactate, and pyruvic acid in the medium were measured after cultured in different condition for 1, 3, 5 days.

RESULTSLow oxygen and low glucose could increase the proliferation of neural stem cells respectively; in addition, the number of neurospheres under both low oxygen and glucose was the most among the four groups. The content of glucose and pyruvic acid in the medium from low oxygen or low glucose condition decreased, while the lactate concentration increased compared with the control group.

CONCLUSIONThe results indicate the neural stem cells prefer grow under the low glucose and low oxygen condition, and that is mainly under going glycolysis to maintain its self-renew ability. This study may provide us a useful clue for application of neural stem cells transplantation.

Animals ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Female ; Glucose ; metabolism ; Glycolysis ; Neural Stem Cells ; cytology ; metabolism ; Pregnancy ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo evaluate and analyze mechanism of Pizhen for the treatment of pain in patients with knee osteoarthritis.

METHODSEighty-five patients (85 knees) were divided into two groups according to random number table method. Among 35 patients in the experimental group, 6 patients were male and 29 patients were female, 1 patient lost follow-up, the average age was (56.65 +/- 4.89) years, and the patients were treated with Pizhen. Among 50 patients in the control group,there were 6 males and 44 females, the average age was (57.54 +/- 4.91) years, and the patients were treated with intra-articular injection with sodium hyaluronate. The VAS score and the data of algometer from the most obvious tenderness around the knee were used to evaluate the clinical effects.

RESULTSThere were significant differences of scores in the experimental group before and after treatment (P<0.01). There were significant differences of scores in the control group before and after treatment (P<0.01). After treatment, there were no statistical differences of VAS score and the data of algometer between experimental and control groups ( P>0.01). There was negative correlation between VAS score and the data of algometer.

CONCLUSIONThe Pizhen is an effective method to treat pain in patients with knee osteoarthritis by relaxing tendon of soft tissue around pain point of knee joint, and relaxing compress and stimulation of nerves.

Acupuncture Analgesia ; Aged ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; physiopathology ; therapy ; Pain Measurement

OBJECTIVETo determine the optimal position of hypoglossal nerve in hypoglossal-facial nerve anastomosis and the eligibility of hypoglossal-facial nerve anastomosis with the cervical loop.

METHODSThe cervical course and adjacent structures of the hypoglossal nerve were observed on 21 adult cadavers. The hypoglossal nerve and facial nerve were taken from 3 fresh specimens, and the number of the fasciculus and the cross-sectional area of the nerve were measured.

RESULTSThe facial nerve trunk were monofascicular with a cross-sectional area of 5.1-/+0.2 (range 4.6-5.7) mm(2). The number of the fasciculus and the cross-sectional areas of the nerve trunk and the fasciculus were 1.6-/+0.8 (range 1-4) mm(2) , 7.5-/+0.7 mm(2) (range 6.8-8.0) mm(2), and 4.7-/+0.6 (4.1-5.5) mm(2), respectively, at the proximal segment of the hypoglossal nerve, 3.6-/+0.5 (1-5) mm(2) , 5.6-/+0.5 (4.9-6.1) mm(2) , and 1.6-/+0.4 (0.9-2.2) mm(2) at the distal segment, and 2.4-/+0.8 (1-3) mm(2), 1.1-/+0.7 (0.6-2.2) mm(2), and 0.5-/+0.3 (0.3-1.2) mm(2) at the cervical loop.

CONCLUSIONThe cervical loop is inadequate for facial nerve anastomosis and the proximal segment is large enough to allow partial harvesting of the hypoglossal nerve for neurotisation of the facial nerve.

Anastomosis, Surgical ; methods ; Cadaver ; Facial Nerve ; anatomy & histology ; surgery ; Humans ; Hypoglossal Nerve ; anatomy & histology ; surgery ; Nerve Transfer ; methods

The anhydrite and gypsum are the main sulfate minerals during evaporation of seawater or lake.They record the information about relative hydrogeology and the composition of mother liquor.Boron is diffluent element, and often occurs in all kinds of evaporites.Presently, the boron isotope has been applied widely in mineral deposits forming, geochemistry and palaeoenvironment.However, there is little research about characteristic of boron isotope in anhydrite and gypsum minerals, because of the low content of boron and micro-solubility in water and hydrochloric acid.This study developed a method of extracting and purifying boron in anhydrite and gypsum by phase transformation and ion-exchange.Firstly, the samples were mixed with ammonium hydrogen carbonate to transform the calcium sulfate to calcium carbonate.And diluted hydrochloric acid (1 mol/L) was added to resolve calcium carbonate.The percent conversion was about 85%in the first stage, and up to complete resolution by repeating this process.Secondly, boron specific ion-exchange resin ( Amberlite IRA 743 ) was used to gather the boron ions fully and further refined the samples with more than 1 μg of boron by anionic and cationic resin mixed by Ion Exchange Ⅱ and Dowex 50 W × 8.Finally, according to the modified method by He, the values of boron isotope were determined by TIMS.The boron content is analytically pure gypsum was 3.501 ± 0.128 μg/g ( n=12 , RSD=3.6%) and the average recovery was 100.47%.Besides, the δ11B value of analytically pure gypsum added with NIST SRM 951 was 17.98‰±0.21‰ (n=3, RSD=1.2%).This method has good repeatability and can meet the requirements of boron isotopic measurement of anhydrite and gypsum.