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.

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.

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

Using universal primer Ty1-copia retrotransposon RT,43 Ty1-copia like retrotransposon RT with high heterogeneity, stop codon mutation and frameshift mutation were amplified by PCR from genomic DNA of Zhejiang Lin'an (C15) and Yunnan Guangnan (A39) of Dendrobium officinale. The length of these sequences varied from 260 to 266 bp, and was rich in AT and consistency ranged from 47.1% to 97.7%. Different c/s-acting regulatory elements induced by low temperature, heat, light, all kinds of plant growth regulating substances and the starting transcription signals, corresponding to CAAT box, TATA box conserved sequences and some other regulatory elements. When being translated into amino acids, ten sequences presented stop codon mutation, five sequences presented frameshift mutation, and thirty-seven sequences presented conserved sequence "SLYGKQ" mutation. Six categories were identified through phylogenic analysis after alignment analyses of their amino acid sequences, and with other plants (eg. Triticum aestivum, Eleocharis quinqueflora) having high homology, which indicated that horizontal transmission of retrotransposon occurred among the plants in the past.
Amino Acid Sequence ; Cloning, Molecular ; Conserved Sequence ; DNA, Plant ; genetics ; Dendrobium ; enzymology ; genetics ; Molecular Sequence Data ; Phylogeny ; RNA-Directed DNA Polymerase ; chemistry ; genetics ; Retroelements ; genetics ; TATA Box ; genetics

Objective To evaluate the safety and efficacy of colour doppler in guiding the percutaneous nephrolithotomy(PCNL) for Staghom calculi.Methods The clinical records of 46 patients with renal calculi who underwent PCNL were retrospectively analyzed.Patients' mean age was 43 years old,and the range of diameter of stone was 3.0 -7.5 cm.Among these cases,29 cases had complete staghorn calculus.One case had isolated kidney stone.And the other 6 patients had open surgery history.Using Colour Doppler guidance,the percutaneous nephrolithotomy for renal calculi was conducted.Results F22 percutaneous channel was successfully established in 55 sides of 46 patients,with the first and the second phase surgeries of 41 and 14 sides respectively.Single-,double- and three-channel PCNL were performed in 38,16,and 1 sides respectively.Abdominal x-ray conducted at 3 -4 days post operation revealed residual stones on 18 sides,with the range of 0.4 -2.0 cm in diameter.Second-phase lithotripsy was conducted on 14 sides of patients.After the first and the second phases of surgery,4 sides having residual stones ranging 0.6 - 1.0 cm,underwent extracorporeal shock wave lithotripsy.Ater the third-phase surgery,the diameters of residual stones were much less than 0.5 cm and patients were treated with medication.The total rate of clearance was 87.0％ (48/55).The duration of surgery was 65 - 160 minutes and 95 minutes on average.One patient having delayed bleeding was cured with selective renal artery embolization.There were no complications such as nephrectomy,deaths,pleural or intestinal damage during the period of study.Patients were followed up for 3 to 18 months.Six of 9 patients with renal insufficiency recovered after surgery.The serum creatinine in the remaining 3 patients ranged 185 -220 μmol/L Conclusion Colour Doppler-guided percutaneous nephrolithotomy for renal calculi is safe and effective.

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.

Objective To explore renal transplantation model in non-human primate cynomolgus monkeys.Methods 50 non-human primates' kidneys were transplanted into the lower part of the abdomen with end-to-side anastomosis of renal artery to aorta and renal vein to inferior vena eava,and with end-to-end anastomosis of ureter to bladder.Results In the 50 cases,1 case death as accident of anesthesia;7 cases with postoperative complications,and all with creatinine sudden rise,after ultrasonic examinations showed that 2 cases with renal vein thrombosis,and 5 cases appeared urinary leakage.All animal models were without surgical infections,and with normal serum creatinine,urine output.Conclusion Non-human primate animal kidney transplantation model establishment method is reliable,but should pay attention to the the surgical technique training,complications prevention.The model is valuable for application in the research of immune tolerance,heterogeneous transplant.

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.

OBJECTIVETo observe the efficacy and safety of acupuncture therapy on moderate and severe allergic rhinitis via the comparison evaluation of western medicine.

METHODSSixty cases of moderate and severe allergic rhinitis were divided into an acupuncture group (30 cases) and a western medicine group (30 cases). In the acupuncture group, the main acupoints included Yingxiang (LI 20), Shangxing (GV 23), Fengchi (GB 20), Quchi (LI 11), Xuehai (SP 10), Feishu (BL 13), Geshu (BL 17), Pishu (BL 20), etc. The supplementary points were selected according to syndrome differentiation. In the western medicine group, budesonide nasal spray and cetirizine tablets were prescribed. All the cases were treated for 8 weeks in the two groups. Separately, before treatment, in 4 weeks and 8 weeks of treatment, the scores of clinical symptoms and physical signs were observed and the clinical efficacy and safety were compared between the two groups.

RESULTSIn 4 weeks and 8 weeks of treatment, the scores of symptoms such as sneezing, nasal discharge, nasal obstruction, nasal itching, eye itching and turbinate hypertrophy, the score of physical signs and total score were all reduced apparently as compared with those before treatment (all P<0. 05). The differences were not significant between the acupuncture group and the western medicine group (all P>0. 05). The total effective rates were 90. 0% (27/30) and 93. 4% (28/30) in the acupuncture group after 4 and 8 weeks of treatment, and were 76. 6% (23/30) and 80. 0% (24/30) in the western medicine group separately, without significant difference in comparison (all P>0. 05).

CONCLUSIONAcupuncture achieves the similar efficacy on moderate and severe allergic rhinitis as western medicine. It is the safe therapy and has no apparent adverse reactions.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Rhinitis, Allergic ; physiopathology ; therapy ; Sneezing ; Treatment Outcome ; Young Adult