Adrenal Hyperplasia, Congenital ; complications ; metabolism ; Body Height ; Growth Disorders ; etiology ; Humans ; Risk Factors ; Steroid 21-Hydroxylase ; metabolism

A single measurement of serum 17alpha-hydroxyprogesterone (17OHP) level can be unreliable because of its marked diurnal variation. We investigated the relationship of serum level of 17OHP with that of androstenedione (AD), which shows a smaller diurnal variation. And we tested whether the responses of these two hormones to low-dose ACTH stimulation are correlated in patients with 21-hydroxylase deficiency. Baseline serum 17OHP and AD levels were measured in 87 patients and a low-dose ACTH stimulation test was performed in 41 patients. The basal 17OHP level correlated positively with the basal AD level independently of sex, type of 21-hydroxylase deficiency, and the time of day of blood sampling (n = 87, R2 = 0.75, P < 0.001). The area under the curve of 17OHP and AD correlated positively with their respective basal levels. The fold-change increase in 17OHP after ACTH injection correlated negatively with the basal 17OHP level, but that of AD did not correlate with the basal AD level. The random serum 17OHP level, used in the clinic, is a reliable guide and a low-dose ACTH stimulation test provides no extra benefit for assessing the treatment adequacy in patients with 21-hydroxylase deficiency.
17-alpha-Hydroxyprogesterone/*blood ; Adolescent ; Adrenal Hyperplasia, Congenital/*diagnosis/drug therapy ; Adrenocorticotropic Hormone/*diagnostic use ; Androstenedione/*blood ; Circadian Rhythm ; Female ; Humans ; Male ; Steroid 21-Hydroxylase/metabolism ; Young Adult

Congenital adrenal hyperplasia is an uncommon diagnosis in routine clinical practice. 21-hydroxylase deficiency, which is its most common subtype, may be diagnosed at birth in a female infant by virilisation or by features of salt wasting in both genders. However, other uncommon subtypes of this condition such as 17-alpha-hydroxylase deficiency, 11-beta-hydroxylase deficiency may present much later in adolescence or adulthood. A high index of suspicion is necessary when evaluating children with hypertension, hypokalaemia, metabolic alkalosis or sexual infantilism.
Adolescent ; Adrenal Hyperplasia, Congenital ; diagnosis ; genetics ; Alkalosis ; diagnosis ; Diagnosis, Differential ; Female ; Humans ; Hypertension ; diagnosis ; Hypokalemia ; diagnosis ; Models, Biological ; Paralysis ; diagnosis ; Sexual Infantilism ; diagnosis ; Steroid 21-Hydroxylase ; metabolism

OBJECTIVETo determine the gene mutation spectrum of patients with 6-pyruvoyltetrahydrobiopterin synthesis deficiency (PTPSD) in Mainland China.

METHODSThe 6-pyruvoyltetrahydrobiopterin synthesis gene lz(PTS)lz was analyzed in 55 PTPSD patients by using PCR-restriction fragment length polymorphism (PCR-RFLP) and direct DNA sequencing. The relationship between the genotype and phenotype was analyzed.

RESULTSEighteen mutations were identified and the detection rate of gene mutation was 95.28%. Four hot-spot mutations, namely P87S (40.57%), N52S(13.21%), D96N(12.26%) and IVS1nt-291A to G(10.38%) were found in this study, and the first three were associated with severe phenotype. The P87L was reported firstly in Chinese patients, and the Q13X, M80T, IVS4nt-2A to G, L93M and K131N were novel mutations.

CONCLUSIONThe P87S, N52S, D96N and IVS1nt-291A to G mutations are the hot-spots mutations of the PTS gene in Chinese PTPSD patients. Using PCR-RFLP technique to screen the mutations in the PTS gene can increase the efficiency of gene diagnosis.

Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; DNA Mutational Analysis ; Female ; Genotype ; Humans ; Male ; Metabolic Diseases ; genetics ; Mutation ; Pedigree ; Phenylalanine Hydroxylase ; genetics ; metabolism ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Pregnancy ; Pterins ; metabolism ; Steroid 21-Hydroxylase ; genetics ; metabolism

OBJECTIVETo establish a rapid method of detecting CYP21 gene mutations.

METHODSFifty Chinese patients with 21-hydroxylase deficiency and some of their families were investigated. Blood samples were obtained for extraction of peripheral blood lymphocytes. A search for restriction sites discriminating between the morbid and the normal in CYP21 gene was made by the computer program DNAssist. PCR-based amplication-created restriction site(PCR-ACRS) was performed at I172N and R356W which are not natural recognition sequence. In addition, I172N and R356W were analysed in five families which conform to the applicability of PCR-ACRS.

RESULTSIn 50 identified 21-hydroxylase deficient Chinese patients, 21 were found to have I172 N (3 were homozygote, 18 were heterozygote); 8 were found to have R356W, all of them were heterozygote. By analysing the families, the findings were consistent with the characteristics of autosomal recessive genetic deficiency.

CONCLUSIONAnalysis of CYP21 gene point mutations using PCR-ACRS is relatively simple, accurate and feasible.

Adrenal Hyperplasia, Congenital ; enzymology ; genetics ; China ; DNA ; genetics ; metabolism ; DNA Mutational Analysis ; methods ; Deoxyribonucleases, Type II Site-Specific ; metabolism ; Female ; Humans ; Male ; Mutation ; Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Sensitivity and Specificity ; Steroid 21-Hydroxylase ; genetics ; metabolism

OBJECTIVETo investigate the clinical and genetic characteristics of 7 patients from 5 families with 17a-hydroxylase/17,20 lyase deficiency (17OHD) and the CYP17A1 mutation in Chinese.

METHODSClinical features and laboratory data were collected from 5 families with 17OHD. PCR direct sequencing was performed to screen the mutation of CYP17A1 gene of the patients. Polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) and sequencing were performed to screen the mutations of CYP17A1 gene in 288 healthy individuals from Shandong province.

RESULTSSeven patients (5 of them were 46,XX; 2 were 46,XY) had typical clinical presentation of sexual infantilism, hypertension and hypokalemia. Hormone profile indicated decreased plasma cortisol and sex hormones, and elevated blood adrenocorticotrophic hormone (ACTH). TAC329AA and H373L in exon 6 and D487_F489del in exon 8 were identified from the patients. One heterozygote for D487_F489del was identified in 288 healthy controls.

CONCLUSIONThe TAC329AA and D487_F489del of the CYP17A1 gene were the most frequent mutations in Chinese with 17OHD.There might be certain frequency of heterozygotes for D487_F489del in Chinese population.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Exons ; Female ; Gene Frequency ; Humans ; Hypertension ; genetics ; Hypokalemia ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation ; Pedigree ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sexual Infantilism ; genetics ; metabolism ; Steroid 17-alpha-Hydroxylase ; genetics ; metabolism ; Steroid 21-Hydroxylase ; genetics ; metabolism ; Young Adult

OBJECTIVEThe major cause of congenital adrenal hyperplasia (CAH) is 21-hydroxylase deficiency, which accounts for 90% - 95% of all cases in most populations. This study was conducted to characterize the molecular basis of the 21-hydroxylase deficiency and to obtain the spectrum of the CYP21 gene mutations in a group of Chinese patients, and analyze the relationship of genotype and phenotype.

METHODSTo detect the distribution of gene mutations in Chinese population samples from 52 patients with 21-hydroxylase deficiency from 51 families were collected, including two parents samples in 30 patients and one parents sample in 10 patients. Blood samples were obtained for extraction of peripheral blood lymphocytes. Polymerase chain reaction (PCR) followed by nesed PCR were used to study the 21-hydroxylase gene (CYP21) mutations. The primary PCR amplified two overlapping CYP21-specific DNA fragments, The product of the nested PCR which used products from the primary PCR was analysed by restriction fragment length polymorphism (RFLP) or amplification-created restriction site (ACRS). All patients were studied by 6 mutations, including P30L, I2g (intron 2 nt 656 c/a-->g splice mutation), E3Delta8nt (exon 3 codon111-codon113 8 bp deletion), I172N, V281L and Q318X.

RESULTSThrough analysis of 52 patients with 21-hydroxylase deficiency, in 5 patients no mutations were detected, in 17 patients only one mutated allele could be characterized, two different mutations were identified in 21 patients, three mutations were detected in 2 patients. Totally, in 73% of alleles the genotypes could be detected. The most common mutation was I2g, which present on 31% affected alleles, then followed by I172N, Q318X, V281L, P30L, E3Delta8nt, accounting for 23%, 14%, 9%, 3%, 2% of all identified mutations respectively, which included multiple mutations accounting for 6%. The most frequent molecular defects of the salt-wasting form were the I2g (45.7%), Q318X (26%). Of the simple virilizing form, the dominant mutations were I172N (40.7%) and I2g (18.5%).

CONCLUSIONSix different mutations were examined in this study, and the detected mutations accounted for 73% affected alleles, in which I2g and I172N were the most common mutations (accounting for 54%). Correlation between genotypes and phenotypes was compatible with the reported data. Two rounds of PCR followed by RFLP or ACRS analysis may provide important information for genetic counseling and for prenatal diagnosis.

Adrenal Hyperplasia, Congenital ; diagnosis ; enzymology ; genetics ; Child ; Child, Preschool ; China ; Family Health ; Female ; Gene Frequency ; Genotype ; Humans ; Infant ; Male ; Phenotype ; Point Mutation ; genetics ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Steroid 21-Hydroxylase ; genetics ; metabolism

OBJECTIVETo analyze CYP21A2 gene mutation in two families with 21-hydroxylase deficiency (21-OHD) and to explore the correlation between genotype and clinical phenotype.

METHODSTwo patients with 21-OHD and their families were investigated. CYP21A2 gene mutation was analyzed by PCR and direct sequencing.

RESULTSThe probands from family 1 and 2 have been respectively diagnosed with simple virilizing and non-classical 21-OHD. Both showed increased baseline serum 17hydroxyprogesterone, testosterone and adrenocorticotropic hormone (ACTH), but had no evidence of salt loss. Computer tomography revealed bilateral adrenal hyperplasia in both patients. After 1 year treatment, both had conceived successfully. DNA sequencing revealed that the proband of family 1 had compound heterozygous mutations for IVS2 13 A>G and Ile172Asn. Her father was heterozygous for Ile172Asn, whilst her mother and brother were heterozygous for IVS213A/C>G. In family 2, the proband was heterozygous for Arg341Trp and Gln318X. Her father, sister and nephew were heterozygous for Arg341Trp, whilst her mother was heterozygous for Gln318X. her brother and niece were non-affected. Carriers of single heterozygous mutations in both families had no clinical sign.

CONCLUSIONIn both families, the disease has been caused by compound heterozygous mutations, for which there has been a good genotype-phenotype agreement. Screening of CYP21A2 gene can facilitate both diagnosis and genetic counseling.

Adrenal Hyperplasia, Congenital ; blood ; enzymology ; genetics ; Adrenocorticotropic Hormone ; blood ; Adult ; Base Sequence ; Child ; Female ; Genotype ; Humans ; Male ; Molecular Sequence Data ; Mutation, Missense ; Pedigree ; Phenotype ; Steroid 21-Hydroxylase ; genetics ; metabolism ; Testosterone ; blood ; Young Adult

OBJECTIVETo gain a primary understanding of the prevalence of 21-hydroxylase deficiency(21-OHD) heterozygote (carrier) among androgen excess women of Chinese Han nationality, compare the molecular genetic changes therein revealed with the results of adrenocorticotropic hormone (ACTH) stimulating test, and assess the carriers' phenotype-genotype correlation.

METHODSEighty-two androgen excess cases and 14 healthy women underwent ACTH stimulating test during the follicular phase. Molecular genetic analysis of CYP21 for 9 common mutations was performed with the method of amplification-created restriction sites.

RESULTSIn androgen excess group, the basal level of F0 (P<0.01), as well as basal 17-OHP0 and the ACTH stimulated concentrations of 17-OHP60 were much higher than controls (P<0.01), and there was no obvious discrepancy in F60 (P>0.05). The net increase of 17-OHP and the ratio of net increase of 17-OHP to net increase of F were also higher than controls (P<0.01). No CYP21 gene mutations were found in control group. Four patients of the androgen excess group were identified as heterozygous carriers of CYP21 mutations. The ACTH stimulating test results from gene normal patients and from carriers overlapped to a certain extent.

CONCLUSIONAmong 82 patients of Chinese Han nationality androgen excess women, 4.9% were 21-OHD heterozygous. The response of serum 17-OHP is not useful for predicting CYP21 gene mutation carrier status. Genotyping is the most reliable method to detect carrier.

Adolescent ; Adrenal Hyperplasia, Congenital ; diagnosis ; ethnology ; genetics ; Adrenocorticotropic Hormone ; Adult ; Asian Continental Ancestry Group ; genetics ; Child ; China ; Female ; Genetic Carrier Screening ; methods ; Genotype ; Humans ; Mutation ; Polymerase Chain Reaction ; Steroid 21-Hydroxylase ; genetics ; metabolism

The present study was performed to investigate the effect of bicyclol on hepatic microsomal cytochrome P450 (CYP) activity, as well as gene and protein expressions in rats after partial hepatectomy (PH). Bicyclol (300 mg x kg(-1)) was given to rats subjected to 70% hepatectomy three times before operation. At 6 and 48 h after PH, blood and liver tissue samples were collected for the measurement of serum alanine aminotransferase (ALT), hepatic microsomal malondialdehyde (MDA) and total hepatic CYP content. The activities of four CYP isozymes were detected with liquid chromatography-mass spectrometry (LC-MS) and the gene and protein expressions were determined by RT-PCR and Western blotting assay. As a result, bicyclol pretreatment markedly inhibited the elevation of serum ALT and hepatic microsomal MDA, and prevented the decrease of total hepatic CYP content in PH rats. In addition, bicyclol significantly attenuated the reduction of CYP2C6 activity and mRNA expression, as well as the reduction of CYP2C11 activity in PH rats. Bicyclol can inhibit the decrease of CYP3A1/2 activity, and up-regulate the mRNA and protein expressions of CYP3A1 and CYP2E1. These results showed that bicyclol pretreatment might ameliorate abnormality in CYP450 isoforms during liver regeneration after PH, and this protective effect was likely due to its anti-oxidative property and enzyme induction.
Alanine Transaminase ; blood ; Animals ; Antioxidants ; pharmacology ; Aryl Hydrocarbon Hydroxylases ; genetics ; metabolism ; Biphenyl Compounds ; pharmacology ; Cytochrome P-450 CYP2E1 ; genetics ; metabolism ; Cytochrome P-450 CYP3A ; genetics ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Cytochrome P450 Family 2 ; Enzyme Activation ; drug effects ; Enzyme Induction ; drug effects ; Hepatectomy ; Male ; Malondialdehyde ; metabolism ; Membrane Proteins ; genetics ; metabolism ; Microsomes, Liver ; metabolism ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Steroid 16-alpha-Hydroxylase ; genetics ; metabolism ; Steroid 21-Hydroxylase ; genetics ; metabolism