Genotype-phenotype relationship in a large cohort of osteogenesis imperfecta patients with COL1A1 mutations revealed by a new scoring system / 中华医学杂志(英文版)

BACKGROUND@#Osteogenesis imperfecta (OI), a heritable bone fragility disorder, is mainly caused by mutations in COL1A1 gene encoding α1 chain of type I collagen. This study aimed to investigate the COL1A1 mutation spectrum and quantitatively assess the genotype-phenotype relationship in a large cohort of Chinese patients with OI.@*METHODS@#A total of 161 patients who were diagnosed as OI in Department of Endocrinology of Peking Union Medical College Hospital from January 2010 to December 2017 were included in the study. The COL1A1 mutation spectrum was identified by next generation sequencing and confirmed by Sanger sequencing. A new clinical scoring system was developed to quantitatively assess the clinical severity of OI and the genotype-phenotype relationship was analyzed. The independent sample t-test, analysis of variance, Mann-Whitney U-test, Chi-squared test, Pearson correlation, and multiple linear regression were applied for statistical analyses.@*RESULTS@#Among 161 patients with OI, 32.9% missense mutations, 16.8% non-sense mutations, 24.2% splice-site mutations, 24.8% frameshift mutations, and 1.2% whole-gene deletions were identified, of which 38 variations were novel. These mutations led to 53 patients carrying qualitative defects and 67 patients carrying quantitative defects in type I collagen. Compared to patients with quantitative mutations, patients with qualitative mutations had lower alkaline phosphatase level (296 [132, 346] U/L vs. 218 [136, 284] U/L, P = 0.009) and higher clinical score (12.2 ± 5.3 vs. 7.4 ± 2.4, P < 0.001), denoting more severe phenotypes including shorter stature, lower bone mineral density, higher fracture frequency, more bone deformity, vertebral compressive fractures, limited movement, and dentinogenesis imperfecta (DI). Patients would not present with DI if the glycine substitutions happened before the 79th amino acid in triple helix of α1 chains.@*CONCLUSIONS@#This presented distinctive COL1A1 mutation spectrum in a large cohort of Chinese patients with OI. This new quantitative analysis of genotype-phenotype correlation would be helpful to predict the prognosis of OI and genetic counseling.

Severe Hyperthyroidism-Heart Failure-Severe Liver Injury:Report of One Case and Literature Review / 中国医学科学院学报

Severe hyperthyroidism can cause the injuries of multiple organs including heart and liver and ultimately be fatal. A 26-year-old young woman admitted to Peking Union Medical College Hospital for severe hyperthyroidism due to irregular use of anti-thyroid drugs. She had heart failure,atrial fibrillation,and severe liver damage at admission. Anti-thyroid drugs were then actively used to treat the primary disease,along with interventions to correct heart failure and control atrial fibrillation. Severe total bilirubin elevation was found during the treatment, which was resolved after the use of glucocorticoid and liver-protective therapy. The patient was regularly followed up after discharge,and the clinical manifestations were good.

A homozygous mutation in TMEM38B causes rare osteogenesis imperfecta type XIV / 基础医学与临床

Objective To investigate the phenotype of a boy with osteogenesis imperfecta(OI)and detect the path-ogenic gene mutation in his family.Methods The clinical data of a uygur ethnic boy was investigated in detail, who suffered from early onset repeated fragile fractures.Bone turnover biomarkers, bone mineral density(BMD) and bone morphology were evaluated.The pathogenic mutations in this patient were investigated by targeted next-generation sequencing and subsequently confirmed by Sanger sequencing.Results Serum β-cross linked C-te-lopeptide of type Ⅰcollagen was elevated.Radiological assessment revealed a generalized osteoporosis in thoraco-lumbar spine,slender long bone with thin cortices.The pathogenic mutations in TMEM38B were detected as follow:a homozygous mutation c.507G>A transition in exon 4,which would generate a new downstream termination codon (p.W169X).His parents were heterozygous carriers of the mutation.Conclusions Mutation in TMEM38B is iden-tified for the first time in a uygur ethnic boy with extremely rare autosomal recessive OI type XIV.The clinical and genetic findings expands our understanding of rare OI induced by TMEM38B mutation.

Association of GNA11 gene polymorphisms with idiopathic hypoparathyroidism and its complications / 基础医学与临床

Objective To investigate the association of GNA 11 gene polymorphisms with idiopathic hypoparathyroidism (IHP) and its complications. Methods Two hundred and three patients with IHP and 209 control subjects were recruited at Peking Union Medical College Hospital from December 1987 to December 2015. The GNA11 gene polymorphisms were selected and genotyped by Sequenom MassArray iPLEX system.Results The minor allele T of rs308060 was associated with an increased risk of IHP in the additive [OR = 2.505(1.005-6.245) , P＜0.05; OR=3.269(1.264-8.458), P＜0.05]and recessive model[OR= 2.727(1.105-6.727), P＜0.05]. Although no significant difference was found in the incidence of intracranial calcification and cataract in IHP patients, the haplotype CTCGCT consisting of minor allele T of rs308060 was correlated with their 24 hours urinary calcium levels (β = 0. 186, P＜0.05). Conclusions The minor allele T of rs308060 in GNA11 means high risk of IHP, and is positively correlated with urinary calcium excretion in IHP patients after treatment with oral calcium and vitamin D analogs supplements.

Clinical features of hereditary distal renal tubular acidosis and SLC4A1 gene mutation / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the clinical features of two families with distal renal tubular acidosis (dRTA) and mutations in the pathogenic gene SLC4A1.</p><p><b>METHODS</b>Family investigation, medical history collection, and measurement of biochemical parameters were performed to analyze the clinical phenotype and genetic characteristics of dRTA. Direct sequencing was used to detect SLC4A1 gene mutations.</p><p><b>RESULTS</b>Three patients in these two families (two of them were mother and son) were diagnosed with dRTA with typical clinical features, including short stature, metabolic acidosis, alkaline urine, hypokalemia, and nephrocalcinosis. SLC4A1 gene analysis showed that all the three patients had a pathogenic missense mutation R589H (c.1766G>A). The child in family 1 had a de novo mutation of SLC4A1, and the child in family 2 had an SLC4A1 gene mutation inherited from the mother, which met the characteristic of autosomal dominant inheritance.</p><p><b>CONCLUSIONS</b>This study reports the R589H mutation in SLC4A1 gene in families with hereditary dRTA for the first time in China. Clinical physicians should perform gene detection for patients suspected of hereditary dRTA to improve the diagnosis and treatment of this disease.</p>

Value of Chloride Clearance Test in Differential Diagnosis of Gitelman Syndrome / 中国医学科学院学报

Objective To investigate the value of chloride clearance test in differential diagnosis of Gitelman syndrome (GS). Methods For patients with hypokalemic metabolic alkalosis and highly suspected GS,clinical data were documented and SLC12A3 gene screening was performed as gold standard to diagnose GS. Hydrochlorothiazide (HCT) test and furosemide (FUR) test were performed according to the standard process. Baseline and maximal increasement of chloride excretion fraction (FECl,the net and relative increase measured as εFECl) were compared between patients and controls to evaluated the reaction to the corresponding diuretics. Receiver operating characteristic (ROC) curve was used to evaluate the sensitivity and specificity of HCT test in GS diagnosis. Results Totally 27 patients and 20 health controls received HCT test. Among those patients,23 were diagnosed with GS genetically. When using the net and relative εFECl to diagnose GS,the areas under the ROC curve were 0.987 (95% CI:0.963～1.000,P<0.001) and 0.984 (95%CI:0.950～1.000,P<0.001),respectively. When a reasonable cutoff value for εFECl was selected,the sensitivity and specificity were both higher than 95%. Eight patients received both HCT test and FUR test. Five of them showed decreased reaction to HCT(net εFECl≤2.86% or relative εFECl≤223%),while normal reaction to FUR.SLC12A3 mutations confirmed their GS. Three patients with blunt reaction to FUR showed normal reaction to HCT,finally they were diagnosed as BS clinically because no SLC12A3 gene mutation was detected. Conclusion Comprehensive application of HCT test and FUR test to evaluate the diuretic reaction can effectively differentiate GS and BS.

Association Between Geranylgeranyl Pyrophosphate Synthase Gene Polymorphisms and Bone Phenotypes and Response to Alendronate Treatment in Chinese Osteoporotic Women / 中国医学科学杂志(英文版)

Objective To investigate the relationship between geranylgeranyl pyrophosphate synthase (GGPPS) gene polymorphisms and bone response to alendronate in Chinese osteoporotic women.Methods A total of 639 postmenopausal women with osteoporosis or osteopenia were included and randomly received treatment of low dose (70 mg per two weeks) or standard dose (70 mg weekly) of alendronate for one year. The six tag single nucleotide polymorphisms of GGPPS gene were identified. Bone mineral density (BMD), serum cross-linked C-telopeptide of type I collagen (β-CTX), and total alkaline phosphatase (ALP) were measured before and after treatment. GGPPS gene polymorphisms and the changes of BMD and bone turnover markers after treatment were analyzed.Results rs10925503 polymorphism of GGPPS gene was correlated to serum β-CTX levels at baseline, and patients with TT genotype had significantly higher serum β-CTX level than those with TC or CC genotype (all P<0.05). No correlation was found between polymorphisms of GGPPS gene and serum total ALP levels, as well as BMD at baseline. After 12 months of treatment, lumbar spine and hip BMD increased and serum bone turnover markers decreased significantly (P<0.01), and without obvious differences between the low dose and standard dose groups (all P>0.05). However, GGPPS gene polymorphisms were uncorrelated to percentage changes of BMD, serum total ALP, and β-CTX levels (all P>0.05).Conclusion GGPPS gene polymorphisms are correlated to osteoclasts activity, but all tag single nucleotide polymorphisms of GGPPS gene have no influence on the skeletal response to alendronate treatment.

Exome sequencing identifies compound heterozygous PKHD1 mutations as a cause of autosomal recessive polycystic kidney disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Autosomal recessive polycystic kidney disease (ARPKD) is a rare inherited disease, which is a disorder with multiple organ involvement, mainly the kidney and liver. It is caused by mutations in the PKHD1 gene. Here, we reported the clinical characteristics of a case with ARPKD and analyze the genetic features of this patient as well as of his father using targeted exome sequencing and Sanger sequencing.</p><p><b>METHODS</b>Genomic DNA was extracted from peripheral blood leukocytes obtained from a patient with ARPKD. The mutations were identified using exome sequencing and confirmed by Sanger sequencing.</p><p><b>RESULTS</b>The patient was diagnosed as ARPKD based on ultrasonography and abdominal computed tomography which showed polycystic changes, multiple calcinosis of both kidneys, and multiple dilated bile ducts of the liver. Compound heterozygous PKHD1 gene mutations A979G and G5935A, which lead to substitution of an asparagine for an aspartate at amino acid 327 (N327D) and a glycine for an arginine at amino acid 1979 (G1979R) respectively, were identified using targeted exome sequencing and confirmed by Sanger sequencing for the patient. In addition, the father of the patient was identified to be a carrier of heterozygous A979G mutation of this gene.</p><p><b>CONCLUSIONS</b>We identified that the compound heterozygous PKHD1 gene mutations are the molecular basis of the patient with ARPKD. Targeted exome sequencing is suitable for genetic diagnosis of single-gene inherited diseases like ARPKD in which the pathogenic gene is a large.</p>

Magnetic resonance imaging findings of pituitary hyperplasia due to primary hypothyroidism / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the clinical and magnetic resonance imaging (MRI) findings of pituitary hyperplasia due to primary hypothyroidism.</p><p><b>METHOD</b>The clinical presentations, laboratory examinations, and MRI findings of 11 patients with pituitary hyperplasia secondary to primary hypothyroidism diagnosed at our hospitals from the beginning of 2008 to the end of 2011 were retrospectively reviewed.</p><p><b>RESULTS</b>The clinical manifestations in 11 patients included growth arrest(7/8), mental retardation (6/8), cold intolerance and fatigue(6/11), slightly increased body weight (6/11), galactorrhea (3/11), paramenia (8/9), precocious puberty companying vaginal bleeding (2/2),and blurry vision (3/11). Laboratory investigations revealed grossly increased thyroid stimulating hormone, decreased thyroxine, and slightly elevated prolactin levels in all cases. Thyroid antibody was positive in six cases. On MRI, pituitary mass were detected a large intrasellar with/without suprasellar extension in all patients,showing the characteristic of symmetric enlargement. Spherical shape was viewed in 5 cases,with the height of (12.22 ± 3.12)mm. In the other 6 cases, the pituitary mass with the shape of calabash extended superiorly to suprasellar area, with a height of(18.95 ± 2.23)mm. The signal of pituitary mass was isointense to grey matter both on T1 weighted imaging and T2 weighted imaging. Bright short T1 signal in posterior lobe of pituitary was visible. Pituitary stalk was detected only in 4 cases from MRI without dislocation, while the width of pituitary stalk was within the normal limit.</p><p><b>CONCLUSIONS</b>Pituitary hyperplasia should be considered when homogenous enlargement of the pituitary gland is found on MRI. The integration of MRI findings, clinical manifestations, and laboratory findings is helpful for the proper identification of the primary endocrine disease and thus avoid misdiagnosis.</p>

Association between vitamin D insufficiency and the risk for gestational diabetes mellitus in pregnant Chinese women / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the association between vitamin D deficiency and risk of gestational diabetes mellitus (GDM) in pregnant Chinese women.</p><p><b>METHODS</b>A nested case-control study was conducted. Clinical and biochemical data were analyzed for 200 subjects with GDM and 200 subjects with normal glucose tolerance (NGT).</p><p><b>RESULTS</b>The median (interquartile range) serum 25-hydroxyvitamin D (25OHD) levels were 22.39 (17.67, 29.38) and 25.86 (19.09, 34.88) nmol/L in the GDM and NGT groups, respectively. Rates of 25OHD deficiency or insufficiency were significantly higher in the GDM group than in the NGT group. Subjects with 25OHD levels <25 nmol/L had a 1.8-fold higher risk of GDM compared with subjects with higher vitamin D levels. In the GDM group, serum 25OHD was independently associated with HbA1c and insulin resistance after adjusting for confounding factors. In the NGT group, serum 25OHD was independently associated with fasting plasma glucose and systolic blood pressure after adjusting for maternal age and other confounding factors.</p><p><b>CONCLUSION</b>25OHD insufficiency is very common in Chinese women. Low 25OHD status may be associated with insulin resistance and act as a risk factor for GDM.</p>

Expression of Ki-67, galectin-3, fragile histidine triad, and parafibromin in malignant and benign parathyroid tumors / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>It is widely recognized that the diagnosis of parathyroid carcinoma (PC) is often difficult because of the overlap of characteristics between malignant and benign parathyroid tumors, especially at an early stage. Our study aimed to investigate the differential expression of Ki-67, galectin-3, fragile histidine triad (FHIT) gene, and parafibromin in PC, parathyroid adenoma (PA), parathyroid hyperplasia (PH), and normal parathyroid (NP) tissues; then to assess these expression values for use in differential diagnosis of malignant and benign parathyroid tumors.</p><p><b>METHODS</b>Data of 15 cases with PC, 19 PAs, and 8 PHs were retrospectively analyzed for their clinical characteristics. The expression of Ki-67, galectin-3, FHIT, and parafibromin were detected via immunohistochemistry in the above-mentioned specimens and 6 NPs as control.</p><p><b>RESULTS</b>Complete loss of parafibromin expression was seen in 9 of 15 (60%) carcinomas, and all normal parathyroid tissues and parathyroid benign tumors stained positive for parafibromin except for one (4%) adenoma. Galectin-3 staining was positive in 11 of 15 (73%) carcinomas, 5 of 19 (26%) adenomas, 1 of 8 (12%) hyperplasias, and 0 of 6 normal tissues. The Ki-67 proliferative index was high in 4 of 15 (27%) carcinomas, 1 of 19 (5%) adenomas, and none of the hyperplasia or normal tissues. FHIT expression did not differ appreciably among the tumor types. The combination of overexpression of galectin-3 or loss of parafibromin increased sensitivity for PC to 87%, while the specificity of both positive galectin-3 and positive Ki-67 could reach 100%.</p><p><b>CONCLUSIONS</b>These data suggested that loss of parafibromin and overexpression of galectin-3 and Ki-67 might help to distinguish parathyroid carcinoma from other parathyroid tumors. And the combination of two or three of these markers might produce better sensitivity and/or specificity for the diagnosis of parathyroid carcinoma.</p>

Surgical treatment of hyperparathyroidism due to parathyroid tumors / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the clinical features of hyperparathyroidism due to parathyroid tumors, and evaluate the efficiency of surgical management.</p><p><b>METHODS</b>Twenty-two patients with hyperparathyroidism resulted from parathyroid tumors were reviewed. The age ranged from 32 to 79 years, 9 males and 13 females. Recurrent laryngeal nerve was routinely exposed, and procedures were performed in normal tissue in initial surgery. Additional selective neck dissection of levels II, III, IV, and VI was taken in the cases with recurrent cancer. Local flaps were used to repair the esophageal defects after resecting tumors. The recurrent laryngeal nerves of 4 cases had to be sacrificed because they were embedded in the tumor tissues despite the nerves had normal function before operation. Prophylactic tracheostomy was performed in 5 cases.</p><p><b>RESULTS</b>Eight cases were identified pathologically as parathyroid carcinoma, of them four with neck metastasis, and 14 cases as parathyroid adenoma after surgery. Their PTH dropped to normal level within two hours after surgery and hypercalcemia disappeared in two days postoperatively. The PTH and serum calcium were in normal range during the follow-up of 12 to 40 months. Recurrence occurred again in two cases in 6 and 8 months after the removal of the recurrent tumor tissues respectively. Esophageal fistula, chylous fistula and dehiscence of sternotomy developed in three cases separately. The tracheostomy was removed in four cases two weeks after operation and in one case six weeks after operation. One patient with parathyroid adenoma died of hypocalcemia about two weeks after operation and another one with recurrent parathyroid carcinoma also died of hypercalcemia 52 months after revised surgery.</p><p><b>CONCLUSION</b>Extended resection of tumor and intraoperative PTH assay were strongly suggested for the managements of both benign and malignant parathyroid tumors.</p>

Diagnosis of thyroid nodules with ultrasound-guided fine needle aspiration: comparison of solid lesion and complex lesion / 中华超声影像学杂志

Objective To compare the efficacy of ultrasound-guided fine-needle aspiration(US-FNA)biopsy in diagnosing solid and complex thyroid nodules with different size. Methods One hundred and seventy-five thyroid FNA biopsies were prospectively performed on 168 patients ranging from 4 to 75 years of age. Sixty-three nodules were surgically excised and the others were clinically followed-up. The cytology diagnoses were categorized into four groups: benign, malignant, suspicious and unsatisfactory. Results There was no significant complication in the all 115 solid and 60 complex thyroid lesions and there were 36and 3 malignant nodules respectively in solid and complex thyroid nodules. The nondiagnostic rates of solid and complex nodules were 7％ and 8％. The accuracy of US-FNA in diagnosing complex thyroid nodules was comparatively equal to that of in solid thyroid nodules. In solid thyroid nodules, the sensitivity and accuracy in ≤1 cm group were similar to that of in ＞1 cm group. Conclusions US-FNA was an accurate and reliable method to diagnose thyroid solid and complex lesions.

Surgical treatment of recurrent parathyroid carcinoma with invasion of the upper aerodigestive tract / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate the factors contributed to the recurrence of parathyroid carcinoma with the invasion of the upper aerodigestive tract and the outcomes of reoperation.</p><p><b>METHODS</b>Six cases reviewed, in which the age ranged from 32 to 79 years old. The initial diagnoses and surgical procedures, the sites and surgical treatment of the recurrent disease, and the chemical markers, such as parathyroid hormone (PTH) and serum calcium, were retrospectively studied. The preoperative PTH levels ranged from 860 to 2830 ng/L. In 4 patients the recurrence diseases were founded in the tracheoesophageal groove, of them one with invasion of the larynx only and one with invasion of the larynx and pharynx in addition to the trachea and esophagus involvement. Selective neck dissection for level II, III, IV and VI was taken in all cases in addition to the removal of the local recurrent diseases. Recurrent laryngeal nerves were so badly embedded in tumor tissue that they were intentionally resected in 4 patients although they were functionally normal before operation. Prophylactic tracheostomy was carried out in 5 cases.</p><p><b>RESULTS</b>PTH level dropped more than 70% of that prior the operation at 10 min after the removal of the tumor-bearing tissues and to normal range within the first 2 hours postoperatively, and hypocalcemia disappeared in 2 days postoperatively. All cases experienced significant improvement in symptoms and signs in the first three days postoperatively. PTH and serum calcium levels were within normal range in 4 cases during the follow-up of 11 to 40 months, while hyperparathyroidism was encountered 8 and 11 months postoperatively in other 2 cases, respectively. Esophageal fistula, chylous fistula and dehiscence of sternotomy developed in 3 cases separately. Of 5 patients with tracheostomy, the tracheostomy tubes were removed two weeks in 4 cases and six weeks in the other one after operation.</p><p><b>CONCLUSIONS</b>Recurrent parathyroid carcinoma even with invasion of the upper aerodigestive tract still has promising surgical outcomes. Both the precise localization of the recurrent diseases and the intraoperative PTH assay are importance to the successful treatment of these diseases.</p>

Benefit of infusions with ibandronate treatment in children with osteogenesis imperfecta / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Osteogenesis imperfecta (OI) is a rare bone disease and its effective treatment is relatively deficient. We tried to assess the effects of new bisphosphonate, ibandronate on children with OI.</p><p><b>METHODS</b>In this open-label, prospective, controlled study, 30 children with OI were enrolled. They received either infusions of ibandronate (2 mg) in every three months or oral calcitriol 0.25 µg daily for 24 months. All patients took 500 mg calcium plus 200 U vitamin D daily together. The endpoints were the change of annual new fracture rate (observed by case history and X ray films of spine), bone mineral density (BMD, measured by dual energy X-ray absortiometry), serum concentration of carboxy-telopeptide cross-links of type I collagen (CTX, bone resorption marker) and alkaline phosphatase (ALP, bone formation marker) during the follow-up.</p><p><b>RESULTS</b>After the cyclic infusions of ibandronate, the annual new fracture rate was significantly decreased from 1.9 to 0.13 time, obviously lower than that of calcitriol group, which decreased from 1.8 to 1.0 time after the treatment (P < 0.001). The significant increase of BMD at the lumbar spine, femoral neck, trochanter, total hip was found in the group of ibandronate by 59.0%, 42.0%, 47.5% and 36.6% in time dependent manner (compared with the baseline, P < 0.001). The increase of BMD in ibandronate group was greater than that of calcitriol group (P < 0.001). The concentrations of ALP and CTX were obviously decreased in ibandronate group, and the reduction of CTX was more significant than that of ALP (P < 0.001). The tolerance of the children to ibandronate was quite well. Mild fever and muscle pain were found in 9 cases within 1 - 3 days after the first infusion of ibandronate, which could relieve after 1 - 2 days without special management.</p><p><b>CONCLUSIONS</b>The benefits of cyclic infusions of ibandronate to children with OI are significant because ibandronate could significantly reduce annual bone fracture rate, increase lumbar and hip BMD, preserve vertebral morphometry of patients through inhibition of bone resorption.</p>

Generalized glucocorticoid resistance accompanied with an adrenocortical adenoma and caused by a novel point mutation of human glucocorticoid receptor gene / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Generalized glucocorticoid resistance syndrome is a rare familial or sporadic condition characterized by generalized, partial, target-tissue insensitivity to glucocorticoids. This syndrome is partially caused by mutations in the human glucocorticoid receptor (hGR) gene. The clinical spectrum of generalized glucocorticoid resistance is broad, ranging from fatigue or no symptoms to severe hypertension with hypokalemic alkalosis. The purpose of this study was to explore the genetic disorder of glucocorticoid resistance syndrome.</p><p><b>METHODS</b>We identified a 56-year-old male patient diagnosed with generalized glucocorticoid resistance syndrome accompanied with an adrenocortical adenoma. This asymptomatic patient referred to Peking Union Medical College Hospital for treatment of his adrenal incidentaloma. Endocrinological evaluation consistently revealed his elevated serum cortisol level. Total RNA was extracted from the patient's peripheral blood mononuclear leukocytes (PBMLs) and entire coding region of hGR alpha was amplified by reverse transcription (RT)-PCR. To confirm the possible mutation identified by sequencing RT-PCR products, genomic DNA sequence of hGR gene from the patient and 50 healthy controls was analyzed by PCR and directly sequencing.</p><p><b>RESULTS</b>A heterozygotic (C→T) substitution at nucleotide position of 1667 (exon 5) in GR alpha gene was found in this patient by sequencing of RT-PCR products of hGR gene. This substitution was also identified at genomic DNA level and it was absent in 100 chromosomes from 50 unrelated health controls. This substitution resulted in a threonine to isoleucine substitution (ACT→ATT) at amino acid 556 in the ligand-binding domain of GR alpha.</p><p><b>CONCLUSION</b>Generalized glucocorticoid resistance in this patient might be caused by a novel heterozygotic mutation in the ligand-binding domain of the GR alpha.</p>

Levels and dynamic changes of serum fibroblast growth factor 23 in hypophosphatemic rickets/osteomalacia / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Hypophosphatemic rickets/osteomalacia is a group of diseases characterised by defective mineralization of bone due to hypophosphatemia and low 1,25-dihydroxy vitamin D. To explore the role of fibroblast growth factor 23 (FGF-23) in the regulation of phosphate homeostasis, we measured the circulating concentrations of this growth factor in healthy individuals and in patients with hypophosphatemic rickets/osteomalacia.</p><p><b>METHODS</b>Nineteen patients with hypophosphatemic rickets/osteomalacia were included in hypophosphatemic group (HP, 12 female and 7 male, mean age was 30 years), and 19 healthy age-matched individuals served as the control group. Full length FGF-23 fragments were measured by two-site enzyme-linked immunosorbent assay.</p><p><b>RESULTS</b>Mean FGF-23 concentrations were significantly higher in the HP group ((87.4 +/- 43.6) pg/ml) compared with the control group ((19.2 +/- 6.16) pg/ml; P < 0.001). In 1 patient with tumour-induced osteomalacia, serum FGF-23 concentrations were 84.1 pg/ml; these concentrations were normalized 2 hours after a hemangiopericytoma resection (7.8 pg/ml). Subsequently, serum 1,25(OH)(2) vitamin D3 concentrations significantly increased from 21.3 pg/ml to 89.3 pg/ml, and serum phosphorus levels were normalized.</p><p><b>CONCLUSIONS</b>Serum FGF-23 concentrations were markedly elevated in patients with hypophosphatemic rickets. FGF-23 plays an important role in the pathogenesis of hypophosphatemic rickets/osteomalacia.</p>

Treatment of hypocalcemia caused by hypoparathyroidism or pseudohypoparathyroidism with domestic-made calcitriol: a prospective and self-controlled clinical trial / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Parathyroid hormone deficiency or resistance may cause hypocalcemia with related symptoms and signs. Lifelong treatment of calcium combined with vitamin D or its metabolites is always necessary for these patients. Here we reported a prospective and open-label trial to investigate the efficacy and safety of domestic-made calcitriol in treatment of hypocalcemia caused by hypoparathyroidism or pseudohypoparathyroidism.</p><p><b>METHODS</b>Twenty-four patients with confirmed hypoparathyroidism or pseudohypoparathyroidism aged (36.5 +/- 11.0) years old were studied. Among them, 16 patients had idiopathic hypoparathyroidism, 2 had pseudohypoparathyroidism and 6 had hypoparathyroidism secondary to cervical surgery. Serum calcium levels were lower than 1.88 mmol/L. Oral calcitriol was administered twice or three times with elemental calcium 1.2 g per day. All patients were followed every 4 weeks throughout the 12-week period. Dose adjustments of calcitriol were based on serum and urinary calcium levels and symptoms of hypocalcemia.</p><p><b>RESULTS</b>Twenty patients were included by the end of this study. Muscular weakness, cramps, extremity paresthesia, Chovestek's sign and Trousseau's sign were relieved in 76.9%, 100%, 94.4%, 93.3% and 78.9% of patients, respectively. Serum calcium, plasma ionized calcium and serum phosphorus levels were (1.54+/-0.25) mmol/L, (0.64+/-0.10) mmol/L and (2.00+/-0.46) mmol/L at baseline, and reached (2.20+/-0.20) mmol/L, (0.95+/-0.06) mmol/L and (1.68+/-0.25) mmol/L (P<0.01) at the 12th week of treatment, respectively. Eighty percent of patients were assessed as effective and 20% as partly effective. Three, four and eight patients had hypercalciuria at the 4th, 8th and 12th week of treatment, respectively, which were reduced by thiazide diuretics. The final dose of calcitriol was (1.09+/-0.50) microg/d.</p><p><b>CONCLUSIONS</b>Calcitriol combined with calcium can be used in treatment of hypocalcemia caused by hypoparathyroidism or pseudohypoparathyroidism effectively and safely. Serum and urinary calcium levels should be monitored during the course of the therapy.</p>

Effect of fibroblast growth factor 9 on Runx2 gene promoter activity in MC3T3-E1 and C2C12 cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Fibroblast growth factor 9 (FGF9), expressed in brain, kidney and developing skeletal tissues, can physiologically inhibit endochondral ossification; but little is known about how FGF9 affects osteoblasts and its detailed regulatory mechanism. Here we examined the effect of FGF9 on the activity of the murine Runt-related transcription factor 2 (Runx2) gene promoter in preosteoblast MC3T3-E1 and premyoblast C2C12 cells.</p><p><b>METHODS</b>Plasmids containing the Runx2 promoter region were transfected into MC3T3-E1 and C2C12 cells and stably transfected cell lines were established. The method of luciferase reporter gene activation was used to examine the effects of FGF9 on the promoter activity.</p><p><b>RESULTS</b>FGF9 (10 ng/ml) increased Runx2 promoter activity in MC3T3-E1 cells. When MC3T3-E1 cells were treated with FGF9 plus the various inhibitors or activator of the intracellular signaling transducation pathways, including 10 micromol/L U0126 (the inhibitor of mitogen-activated protein kinase kinase), 10 micromol/L SB203580 (the inhibitor of p38/mitogen activated protein kinase), or 1 micromol/L C6 ceramide (an activator of mitogen activated protein kinase), the luciferase expression did not change significantly compared with that of the cells treated with FGF9 only. However, when C2C12 cells were treated with 10 ng/ml FGF9, Runx2 gene promoter activity first decreased and then increased over a period of 1 to 5 days. Among the above inhibitors, only U0126 (10 micromol/L) completely blocked the effects of FGF9 on Runx2 gene promoter activity.</p><p><b>CONCLUSIONS</b>Our data showed that FGF9 can affect Runx2 gene promoter activity in MC3T3-E1 and C2C12 cells. The action of FGF9 appears to depend partly on the mitogen-activated protein kinase kinase/mitogen-activated protein kinase pathways in C2C12 cells.</p>

RET gene cys 634 trp mutation in a multiple endocrine neoplasia type 2A kindred / 中国医学科学院学报

<p><b>OBJECTIVE</b>To identify the genotype of RET gene in one multiple endocrine neoplasia type 2A (MEN2A) kindred.</p><p><b>METHODS</b>Genome DNA was extracted from peripheral blood leucocytes. The DNA sequence of gel-purified polymerase chain reaction (PCR) products was determined with the previously reported 6 pairs of primers of PCR amplification of 10, 11, 13, 14, 15, and 16 exons of RETgene.</p><p><b>RESULTS</b>No abnormalities were found in exon 10, 13, 14, 15, and 16. C to G replacement in nucleotide 14 996 of exon 11 was identified in DNA samples obtained from both peripheral blood of 2 affected brothers. This missense point mutation arisen in heterozygosity and caused a substitution of Cys to Trp residue at codon 634 ( Cys 634 Trp) in RET protein.</p><p><b>CONCLUSION</b>The genotype of the family is identified as Cys 634 Trp substitution of RET gene.</p>