BACKGROUND: Phenylketonuria is caused by gene mutation of phenylalanine hydroxylasel (PAH), which is mainly induced by permutation, short segments and insertion of base.OBJECTIVE: To evaluate the gene mutation of phenylalanine hydroxylasel in phenylketonuria in Hui nationality.DESIGN: Open study.SETTING: Urumqi General Hospital of Lanzhou Military Area Command of Chinese PLA; Capital Pediatrics Institute.PARTICIPANTS: A boy of Hui nationality in China and aged 3.1 years was selected in this study. The boy had intellect hysteresis in his one year and received medical treatment in his three years, while he was diagnosed as cerebral paralysis. After repeatedly inefficient treatment, he was hospitalized in our hospital on December 13, 2004. Iron sesquichloride in urine was strongly positive and concentration of serum phenylalanine was 1 680 μmol/L; therefore, he was diagnosed as the typical phenylketonuria.METHODS: 5 mL venous blood was selected from the boy and his parents, respectively, and anticoagulated with EDTA-Na2. DNA in gene group was extracted by using typical phenol/chloroform method. In addition, polymerase chain reaction (PCR) primer sequence of extron 7, 6, 11, 3, 12 and 5 of PAH gene was designed based on references. And then, PCR products were detected with 2% agarose gel electrophoresis. 5 μL PCR products were mixed with the same volume of degenerated buffer solution, degenerated at 97 ℃ for 5 minutes, put in iced bath and performed with 80 g/Lnon-degenerated polyacrylamide gel electrophoresis. After that, the products were dealt with sliver staining routinely, and single strand DNA banding patterns were analyzed and recorded. ABI377 automatic sequenator (PE Company) was used to detect PCR sequence and purify PCR product in Shanghai Boya Biotechnology Company.MAIN OUTCOME MEASURES: Iron sesquichloride in urine, concentration of serum phenylalanine and mutant gene types of phenylalanine hydroxylase.RESULTS: Extron 7, 6, 11, 3, 12 and 5 of PAH gene were analyzed in the boy and his parents. The results demonstrated that SSCP electrophoresis in extron 6 was different from that in the normal control group. Site of electrophoresis strip of his father was coincident with that of his mother, but different from that of the boy. Sequencing results indicated that point mutation (cytosine replaced by thymine), which was a R176X mutant heterozygote, occurred at the 526th site of cDNA of phenylalanine hydroxylase gene in his parents; however, two chromosomes of the boy had mutation at the same site, which was R176X mutant homozygote.CONCLUSION: Mutation of R176X homozygote of phenylketonurea is firstly reported in Hui nationality in China.

Objective To study the potential efficacy of transplanted bone marrow-derived mesenchymal stem cells (MSCs) in treating and repairing the acute lung injury in animal models. Methods MSCs were isolated from mouse bone marrow, cultrued and amplified in vitro. The lipopolysaccharide (LPS) was inhaled through postnasal tract to cause acute lung injury in mice and the MSCs labeled by Brdu were administrated via vein into the mice. The migration and differention of the cells were identified by immunostaining and double immunostaining. The pathological changes, pulmonary edema index and the content of IL-1β in lung homogenate were used to accese the therapeutical effect of MSCs. Results The cultured MSCs dispalyed a positive CD44 and a negative CD34. The Brdu-labeled cells were detected in the lungs of the recipient 4 days after transplantation, indicating its origin of MSCs. Theses cells also exhibited characteristics of aveolar epithelials, expressing the cytokeratin-the marker of epithelium. Compared with the injuried ones, the mice treated with MSCs showed a decreased pulmonary edema in-dex and IL-1β content in the lung homogenate. Conclusion These data suggest a therapeutical effects of MSCs in treating and repairing the mouse acute lung injury.

OBJECTIVETo identify the mutation spectrum and the distribution of minihaplotypes (STR/VNTR) of phenylalanine hydroxylase (PAH) gene and explore the correlations between genotype and phenotype of patients with phenylketonuria (PKU) in Beijing area of China.

METHOD(1) Fifty cases with PKU were involved in this study. PKU was identified by the Neonatal Screening Center of Beijing. All 13 exons and their flanking intronic sequences of PAH gene of these patients were amplified and then subjected to SSCP analysis and direct sequencing. (2) The distribution of polymorphic locus of short tandem repeat (STR) and variable number tandem repeat (VNTR) was analyzed by PCR and denaturing gel electrophoresis. (3) The correlations between genotype and phenotype were studied by analysis of the matching rate between the expected and observed phenotypes. The predicted phenotype was determined on the basis of the sum of the assigned values of the two mutant alleles.

RESULTS(1) A total of 34 different mutations were detected with the relative frequency of 95% among 50 PKU patients. The prevalent mutations in this study were: R243Q (20%), EX6-96A > G (11%), Y356X (9%), and V399V (7%). The next common mutations were R111X (5%), R413P (5%), R252Q (3%) and A434D (3%). Thirty-four detected mutations were distributed throughout the whole PAH gene, except exon 1, 8 and 13. Exon 7 and 11, with the mutant rate 34% and 19% respectively, seemed to be the hot mutant areas/regions of PAH gene. (2) The minihaplotypes (STR/VNTR) of 34 mutations were identified in this research. The STR and VNTR showed 8 and 3 alleles, respectively. Among them, 244 bp (44%) and 240 bp (34%) were the prevalent STR alleles. Meanwhile, the VNTR3 (83%) was the most common VNTR allele in PKU patients. (3) A better consistency (81.5%) between expected and observed phenotypes was revealed by analysis of correlation between genotype and phenotype. Especially in classic PKU, the consistency rate was up to 87.5%.

CONCLUSION(1) The frequency distribution of common PAH gene mutations in Beijing region was close to that of Tianjin and Yunnan regions, while it was different from that of Southern regions of China, such as Guangzhou, especially Taiwan. The PAH mutation with a highly heterogeneous trait was also demonstrated in this study. (2) STR and VNTR minihaplotype will prove helpful to trace the origins of PAH mutations and to analyze the genetic drift. However, the most minihaplotypes of the STR/VNTR are similar, so it is necessary to associate some other polymorphic loci with the STR/VNTR minihaplotype to analyze the different mutations. (3) The fact that a better consistency existed between phenotypes and genotype with most PKU patients suggested that the study of the genotype of PKU patients would be helpful to the individualized treatment and to genetic counseling for their families.

Alleles ; China ; epidemiology ; Genetic Association Studies ; Genotype ; Humans ; Infant, Newborn ; Introns ; Mutation ; Phenotype ; Phenylalanine Hydroxylase ; genetics ; Phenylketonurias ; epidemiology ; genetics ; Polymorphism, Genetic

A multiplex real-time PCR was developed to detect ctxA of Vibrio cholerae, gyrB and tdh of Vibrio parahaemolyticus simultaneously. The multiplex real-time PCR were evalidated by detection for the three genes in 47 toxigenic V. cholerae O1 and O139 strains (ctxA+; O1=3, O139=44), 25 non-toxigenic V. cholerae strains (ctxA-; O1=12, O139=6, non-O1 and non-O139=7), 116 V. parahaemolyticus strains with or without tdh (73 or 43) and 9 other bacteria strains. The specificity and sensitivity of the multiplex real-time PCR in detection for the ctxA and the tdh genes in the strains tested were both 100.0%, compared to the results by routine PCRs. In the detection for V. parahaemolyticus specific gyrB using the multiplex real-time PCR, all of 116 V. parahaemolyticus strains were positive, and 9 other strains and 72 V. cholerae strains were all negative. The multiplex real-time PCR is a sensitive, specific and quick assay not only for detecting virulence genes of V. cholerae and V. parahaemolyticus but also for identifying V. parahaemolyticus at species level. In addition, two real-time PCRs for detection of V. parahaemolyticus virulence genes trh1 and trh2 were also developed.

Objective:To investigate association of the new polymorphism G352A in the dopaminetransporter gene(DAT1)exon 15 with attention deficit hyperactivity disorder(ADHD)in Han Chinesechildren.Methods:The new mutant polymorphism G352A in the dopamine transporter gene(DAT1)ex-on 15 was found by the fluorescently-labeled dye-terminators assay.The study samples were comprised of337 ADHD children,207 unrelated controls and 201 integrated ADHD trios(included proband and bio-logical parents).Associations of polymorphisms with ADHD and its subtypes were examined by:(i)comparing cases and controls;and(ii)using family-based association study in transmission-disequilibri-um test(TDT).Results:The allele frequencies at the DAT1 G352A locus in the control samples were79.5% for 352G and 20.5% for 352A respectively.Association studies revealed no association betweenG352A in exon 15 of DAT1 and ADHD.But after a stratification by gender,there was possible associationbetween G352A and ADHD girls:the 352G allele had a tendency to be preferentially transmitted toADHD girls.Conclusion:There is no association between G352A,the new polymorphism,in exon 15 ofDAT1 and ADHD.The 352G allele has a tendency to be preferentially transmitted to ADHD girls,but thefindings require replication before drawing a definitive conclusion.

Objective The aim of this study is to evaluate the efficacy and safety of preoperative CT-guided hardening agent localization.Methods From December 2010 to January 2012,27 patients with 29 solitary pulmonary nodules who had undergone CT-guided hardening agent localization and video-assisted thoracoscopic surgery (VATS) were studied.Results All cases were underwent CT-guided hardening agent localization successfully,and no patient had serious complication that required any intervention.The diameter of nodules ranged from 3 to 21 mm as measured by CT[mean (11.27 ± 6.32) mm].The distance between the center of nodule and visceral pleural ranged from 4 to 38 mm[mean (14.45 ± 4.32) mm].Conversion from VATS to thoracotomies was not necessary during the diagnostic resection procedure nodules.29 solitary pulmonary nodules underwent thoracoscopic wedge resection,and no intra-or postoperative mortality or morbidity was recorded.Conclusion CT-guided hardening agent localization before video-assisted thoracoscopic solitary pulmonary nodule resection is a safe and effective procedure for accurate diagnosis and resection of indeterminate solitary pulmonary nodules.

We report one case of pediatric acute myeloid leukemia type 2 (AML-M2) who presented with karyotypic aberration of trisomy 21 with the t(5;ll) chromosomal translocation.The patient achieved complete remission after two cycles of chemotherapy of daunorubicin,cytarabine and etoposide.Then,follow-up cytogenetic analysis from bone marrow cell cultures demonstrated a normal karyotype of 46,XY.After 9 years,the patient relapsed and the karyotypic abnormalities of trisomy 21 with t(5;ll) reappeared.It was concluded that trisomy 21 with t(5;11) is a new unfavorable cytogenetic aberration in AML-M2.

Objective To investigate whether the macromolecular materials could enter cerebrospinal fluid and brain tissues in craniotomy with incision or non-incision of dura and arachnoid. Methods Adult male SD rats were randomly divided into three groups according to the random number table. The dura and arachnoid of rats in group A were cut open during craniotomy after general anesthesia; epidural craniotomy was done in rats in group B after general anesthesia; rats in group C (control group) were only generally anesthetized. All the rats were injected with Evans blue, a tracer used to detect the results, half an hour before each time point (1,3, 6, 12, 24, 72 hours and 1 week) via vein. The rats were executed at each time point to obtain the specimens of brain. The content of Evans blue in brain tissue was measured by fluorescence spectrophotometer for statistical analysis. The water content in the brain tissue was measured in a part of rats selected in groups A and B preoperatively and at postoperative 3 and 27 hours. Results It was found that some regions of the brain tissue were stained light blue in group A at 1,3, 6 and 24 hours. The blue was much lighter in brain tissue obtained at 72 hours in group A, and no blue stained at 1 week in group A . The contents of Evans blue in the brain tissues of rats in group A at 1,3, 6, 12, 24, 72 hours and 1 week were (18.07±1.25) μg/ml, (36.21±0.78) μg/ml, (25.73±1.14) μg/ml, (16.53±0.84) μg/ml, (23.34±1.91) μg/ml, (43.34±2.25) μg/ml and (25.27±1.88)μg/ml respectively, which were significantly higher than (3.15±0.45)μg/ml, (3.36±0.33)μg/ml, (2.98±0.54)μg/ml, (3.47±0.55)μg/ml, (3.54±0.37) μg/ml, (2.88± 0.42) μg/ml and (2.85±0.22) μg/ml respectively in group B and (2.97±0.37)μg/ml in group C (P<0.01). There was no significant difference in water content in brain tissue before and after operation (P>0.05). Conclusion After craniotomy with incision of dura and arachnoid, some macromolecular materials can enter the subarachnoid space and the brain parenehyma through blood-brain barrier of the wound of the scalp if the dura is sutured loosely.

Objective To analyze the ultrasonic features of thyroid microcarcinoma (TMC) and the causes of misdiagnosis. Methods The ultrasonic features including shape, margin, echogenecity, microcalcification, vascularity and lymphadenopathy were analyzed retrospectively in 26 pathologically-proven TMC patients. Results In 26 cases, 11 cases were diagnosed correctly before operation (11/26, 42.31%), 12 cases were misdiagnosed (12/26, 46.15%) as adenoma or benign nodule, and 3 cases were missed diagnosed (3/26, 11.54%). Among the 23 cases detected on ultrasound, 21 cases were solid and hypoechoic (21/23, 91.30%);19 cases were ill-defined (19/23, 82.61%);12 cases were taller than wide in shape (12/23, 52.17%); 14 cases had microcalcification (14/23, 60.87%); 7 cases showed central or peripheral blood flow signals (7/23,30.43%) with arterial resistance index>0.70 in 3 lesions and<0.70 in 4 lesions. Conclusions Several ultrasonographic features are helpful in identiifcation of TMC, including hypo/iso-echogenecity, ill-deifned margin, taller-than-wide shape, microcalciifcation, arterial signals with high resistance index, and abnormal lymphadenopathy. Moreover, for cases with multiple lesions, to the potential co-existence of benign and malignant lesions should be considered.