Variation Study of Steady-State trough Concentration of Adalimumab in Serum of Patients with Ankylosing Spondylitis / 中国药学杂志

OBJECTIVE: To establish an enzyme-linked immunosorbent assay (ELISA) method for determining the concentration of adalimumab in human serum to determine the steady-state trough concentration of adalimumab in patients with ankylosing spondylitis (AS) and to study inter- and intraindividual variation. METHODS: Human tumor necrosis factor α (hTNF-α) was coated on the solid phase microplate as the capture antigen, and the sample to be tested was added. The goat anti-human IgG-HRP was used as the detection antibody and TMB was used for color development. The absorbance at 450 nm was finally determined. Forty-seven patients with AS treated with standard dose of adalimumab injection were included. Blood samples of steady-state after 12 to 24 weeks′ treatment were collected to determine trough drug concentrations. RESULTS: The quantitative range of the method was 0.63-20.00 μg•mL-1. The inter-assay and intra-assay precisions were not more than 15% and the average recovery was <110%. The median serum concentration of 47 patients was 10.15 μg•mL-1 (IQR:4.36~13.31 μg•mL-1) and ranged from 0.00-23.71 μg•mL-1 with an inter-individual variation of 63%; the median variation of intra-individuals within 43 patients was 11% (IQR: 8%-17%). CONCLUSION: This method is stable and is suitable for the detection of clinical samples. Pronounced inter- but not intraindividual variation was observed in adalimumab trough levels in steady state. The "reaction therapeutic drug monitoring" based on the steady-state trough concentration will facilitate the adjustment of the dosing regimen.

Advances in Therapeutic Drug Monitoring of Monoclonal Antibody Drugs / 中国药学杂志

Monoclonal antibody (mAb) drugs currently are an important field in drug development and playing a wide role in the treatment of autoimmune diseases and tumor. However, interindividual variability of therapeutic response and loss of response during treatment become serious problems in clinic, and the underlying mechanisms are believed to be closely related to drug exposure in patients and immunogenicity. Therapeutic drug monitoring (TDM) of mAb drugs has the potential to guide more effective dosing in individual patients and it has been proved to be helpful when making treatment decisions particularly for tumor necrosis factor inhibitors. This review is to summarize the pharmacokinetic profiles and exposure-response relationship of mAb drugs, so as to provide theoretical basis and research direction for the application of TDM in mAb drugs.

Mutation analysis and prenatal diagnosis of families affected with Duchenne and Becker muscular dystrophy / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect potential mutations for probands from families affected with Duchenne/Becker muscular dystrophy (DMD/BMD), and to carry out prenatal diagnosis through identification of female carriers.</p><p><b>METHODS</b>A total of 43 DMD/BMD families were recruited. Multiplex PCR was used to analyze 18 exons within hotspots for DMD gene deletions. Multiplex ligation-dependent probe amplification (MLPA) was used to detect potential deletions and duplications of DMD gene for 43 patients and 36 females from 32 families. Prenatal diagnosis was performed for 27 families.</p><p><b>RESULTS</b>Deletional mutations were detected in 26 patients with multiplex PCR. In addition, MLPA has detected 3 deletions and 6 duplicational mutations, and the ranges of mutations were all determined. Among 36 female members, 18 were determined as carriers of deletional mutations, 10 were excluded as mutation carriers. The status of remaining 8 could not be determined. For prenatal diagnosis, 3 out of 18 male fetuses were diagnosed as patients and 1 female fetus was identified as carrier.</p><p><b>CONCLUSION</b>MLPA is an accurate and reliable method for detecting deletional/duplicational mutations of DMD gene as well as for prenatal diagnosis and detection of female carriers.</p>

Molecular genetics study of ED1 gene for two X-linked hypohidrotic ectodermal dysplasia families / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To provide genetic diagnosis and counseling for patients from two families affected with X-linked hypohidrotic ectodermal dysplasia.</p><p><b>METHODS</b>Potential mutation of the ED1 gene was screened by DNA sequencing. For family 1, multiplex ligation-dependent probe amplification (MLPA) analysis and haplotyping of ED1 gene were also carried out for prenatal diagnosis.</p><p><b>RESULTS</b>For the patient from family 1, deletion of the exon 1 of the ED1 gene and 2 short tandem repeat(STR) sites (DXS8269 and DXS1422) were detected. His daughter was carrier of the deletion. Upon prenatal diagnosis, the fetus was confirmed to be a normal male, for whom the haplotype of ED1 gene has differed from that of the proband. In family 2, a c.463C>T mutation in exon 3 of the ED1 gene was detected in the proband, whose mother was heterozygous for the same mutation.</p><p><b>CONCLUSION</b>The deletion (exon 1) and missense (R155C) mutation in ED1 gene have probably underlied the disease in the two families. During prenatal diagnosis, it may be necessary to obtain precise results through combining mutation detection and haplotype analysis of the ED1 gene.</p>

Mutation analysis and prenatal diagnosis of EXT1 gene mutations in Chinese patients with multiple osteochondromas / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Multiple osteochondromas (MO), an inherited autosomal dominant disorder, is characterized by the presence of multiple exostoses on the long bones. MO is caused by mutations in the EXT1 or EXT2 genes which encode glycosyltransferases implicated in heparin sulfate biosynthesis.</p><p><b>METHODS</b>In this study, efforts were made to identify the underlying disease-causing mutations in patients from two MO families in China.</p><p><b>RESULTS</b>Two novel EXT1 gene mutations were identified and no mutation was found in EXT2 gene. The mutation c.497T > A in exon 1 of the EXT1 gene was cosegregated with the disease phenotype in family 1 and formed a stop codon at amino acid site 166. The fetus of the proband was diagnosed negative. In family 2, the mutation c.1430-1431delCC in exon 6 of the EXT1 gene would cause frameshift and introduce a premature stop codon after the reading frame being open for 42 amino acids. The fetus of this family inherited this mutation from the father.</p><p><b>CONCLUSIONS</b>Mutation analysis of two MO families in this study demonstrates its further application in MO genetic counseling and prenatal diagnosis.</p>

Molecular genetic study of MECP2 gene for a patient with typical Rett syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To provide genetic diagnosis and counseling for a 2-year-old girl with typical Rett syndrome through analyzing the methyl-CpG binding protein 2 (MECP2) gene.</p><p><b>METHODS</b>Potential mutation of the MECP2 gene was screened by DNA sequencing and multiplex ligation-dependent probe amplification (MLPA) analysis of members of the family as well as normal controls. Lymphocyte culture for karyotype analysis was carried out for the patient to exclude chromosomal abnormalities.</p><p><b>RESULTS</b>The karyotype of the girl was normal. No variation of the MECP2 gene was detected in the patient by direct sequencing. A heterozygosis variation, c.1072G>A in exon 4 of the MECP2 gene was detected in a normal female control, which was not found in other controls. The son and daughter of the female control were respectively heterozygous and homozygous carriers of the same mutation. By MLPA analysis, a heterozygosis deletion of exon 3 and part of exon 4 was detected in the patient. cDNA amplification and sequencing confirmed the presence of a 1176 bp deletion (c.27-1202del1176). The same deletion was not detected in the parents.</p><p><b>CONCLUSION</b>A large deletion in MECP2 gene was detected with MLPA in a patient featuring typical Rett syndrome. The same deletion was missed by sequencing analysis. With cDNA sequencing, the breakage point of the mutation can be mapped precisely.</p>

Mutational analysis in a family with X-linked spondyloepiphyseal dysplasia tarda / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect the mutation of the SEDL gene in an X-linked spondyloepiphyseal dysplasia tarda (SEDL) family.</p><p><b>METHODS</b>Two patients and three females of the X-SEDL family were detected using reverse transcriptase PCR (RT-PCR) and sequence analysis.</p><p><b>RESULTS</b>A G209A mutation of SEDL gene was detected in the cDNA sequences of the patients, which was confirmed by sequence analysis of the exon 4 of the SEDL gene. The daughter of the proband was a carrier of the mutation.</p><p><b>CONCLUSION</b>Since the SEDL gene is relatively small, sequence analysis of cDNA of the SEDL gene was possible after extraction of total RNA followed by RT-PCR. Mutations in the open reading frame can be detected y by cDNA sequencing. It was relatively more rapid and direct than amplifying and detecting the exons one by one.</p>

Evaluating the applicability of MoM in second trimester maternal serum to screening for Down's syndrome in Jiangsu province / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the applicability of MultiCalc software to prenatal screening of Down's syndrome in Jiangsu province, China.</p><p><b>METHODS</b>The gestational age-specific median of maternal serum marker was calculated by means of regression method. Regression functions for adjustment of Multiple of the Median (MoM) by weight were established for our own population.</p><p><b>RESULTS</b>Before the adjustment by weight, the average level of alpha fetal protein(AFP) was 16% higher and the free beta-human chorionic gonadotrophin (beta-hCG) was 14% higher than those of the Caucasian in MultiCalc software respectively. But when the AFP and free beta-hCG results were converted to weight-adjusted MoM levels, the values were 0.99 and 1.02 respectively. The median of MoM of AFP and the free beta-hCG were 1.00 through the regression model of gestational age and weight adjustment.</p><p><b>CONCLUSION</b>There was no difference of average weight-adjusted MoM levels between the Jiangsu population and the Caucasian, and the MultiCalc software was applicable to maternal serum screening for Down's syndrome of Jiangsu.</p>

Protective effect of curcumin on oleic-induced acute lung injury in rats / 中国中药杂志

To investigate the effect of curcumine on acute lung injury induced by oleic acid in rat and the possible mechanism of action. The rats were divided into 6 groups randomly: normal group, control group, curcumine groups (5, 10, 20 mg x kg(-1)) and dexamethasone group (1 mg x kg(-1)). During the experiment, acute lung injury was induced by oleic acid in rat. The changes of dynamic lung compliance were recorded by anrise 2005 pulmonary function test apparatus, light microscope was used to examine histological changes and lung index as well as wet to dry weight ratio was calculated by weighting method. Lung vascular permeability and protein level in BALF were detected by ultraviolet spectrophotometry, and the concentrations of TNF-alpha, IL-6 and IL-10 in BALF were measured by enzyme linked immunosorbent assay (ELISA). The result showed that the changes of pulmonary compliance were inhibited and pulmonary function was improved by curcumine. The OA-induced elevation of lung index was restrained, as well as wet to dry weight ratio, lung vascular permeability, protein level, TNF-alpha (250.4 +/- 21.6 vs. 172.53 +/- 14.88, 122.2 +/- 10.98, 108.69 +/- 3.39) ng x L(-1), IL-6 (763.6 +/- 88.33 vs. 207.41 +/- 15.55, 172.13 +/- 21.91, 142.92 +/- 4.32) ng x L(-1) in BALF in curcumine groups, IL-10 (98.90 +/- 2.99 vs. 208.44 +/- 16.30, 218.43 +/- 6.23, 252.70 +/- 20.58) ng x L(-1) in BALF was increased, respectively significantly. Light microscope findings shown that the impairment in curcumine groups was far less severe than that in model groups. Pretreatment of curcumine showed beneficial effect on acute lung injury induced by oleic acid in rats. The mediation of both proinflammatory factor and anti-inflammatory factor by curcumine may be involved in mechanism of action of curcumine effects.

Effect of resveratrol on chronic obstructive pulmonary disease in rats and its mechanism / 药学学报

The purpose of this study is to establish COPD animal model by intra-tracheal instillation of bleomycin (BLM) once and exposure to cigarette smoke for continuous 27 d, and to observe the effects of the inhalation on the model. At the 29th day, blood samples were taken from cervical artery for blood-gas analysis and parameters of lung function were recorded. Bronchoalveolar lavage fluid (BALF) was collected to measure intercellular adhesion molecule-1 (ICAM-1) concentration. The results showed that atomization inhaled resveratrol could alleviate rat COPD lung injury accompanied by amelioration of pathological changes, increase the ratio of forced expiratory volume in 0.3 s (FEV0.3) and forced vital capacity (FVC), and decrease the ICAM-1 level in BALF. The ultimate reduction of inflammatory factors was involved, at least in part, in the mechanism of resveratrol effects.

Detection and related analysis to chromosome 22q11 microdeletion in patients with congenital heart diseases / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To ascertain 5 short tandem repeat (STR) markers as qualified tools for detecting chromosome 22q11 deletion and to understand the prevalence and clinical importance of the deletions in patients with congenital heart diseases (CHD) from Chinese Han population.</p><p><b>METHODS</b>The authors selected 5 new tetranucleotide repeat markers, 22D_4_1, 22D_4_2, 22D_4_3, 22D_4_4 and D22S873 located in the proximal region of chromosome 22q11 deletion. One hundred and sixty-three unselected CHD patients and their unaffected parents were analyzed by genotyping of these new tetranucleotide STR markers to detect 22q11 deletion. With fluorescence in situ hybridization (FISH, LSI dual color DNA probe), the deletion status was confirmed in all patients with deletions and some patients without deletions.</p><p><b>RESULTS</b>The heterozygosity of these STR markers in normal population was more than 0.7, except for 22D_4_1 and 22D_4_2 that were 0.65 and 0.52 respectively. Twelve cases of 163 CHD patients (7.36%) had the deletions at chromosome 22q11. The deletions were confirmed in 9 of 12 patients by FISH, except for 2 cases who had unique nested deletion and 1 case who had nested distal deletion. One hundred and ten patients were associated with ventricular septal defect (VSD); and 9 (8.18%) of these cases had microdeletion. Twenty-one patients were associated with tetralogy of Fallot (TOF); and 3 (14.3%) of these cases had microdeletion.</p><p><b>CONCLUSION</b>This study demonstrated that genotyping of 5 STR markers was a useful mean of detecting 22q11 microdeletion in clinical diagnosis owing to its rapid experimental procedure, cost effectiveness and high resolution. 22q11 deletion was common in CHD patients, particularly in VSD and TOF patients, from Chinese Han population.</p>