RNA methyltransferase is responsible for transferring methyl and resulting in methylation on the bases or ribose ring of RNA, which existed widely but mostly remains an open question. A recombinant protein PH1948 predicting RNA methyltransferase from Pyrococcus horikoshii OT3 has been crystallized. The crystals of selenomethionyl PH1948 belong to space group C2, with unit-cell parameters a=207.0 A, b=43.1 A, c=118.2 A, b=92.1 degrees , and diffract X-rays to 2.2 A resolution. The V(M) value was determined to be 2.8 A3/Da, indicating the presence of four protein molecules in the asymmetric unit.
Crystallization ; methods ; Molecular Conformation ; Pyrococcus horikoshii ; enzymology ; X-Ray Diffraction ; tRNA Methyltransferases ; analysis ; chemistry

Objective To investigate the changes of the constitution and its ratio of collagen fiber in the process of masseter reattachment following different osteotomies of the prominent mandibular angle so as to offer guidance for the resection of mandibular angle. Methods Sixteen adult goats were randomized into four groups. In group A we performed unilateral curved osteotomy of the mandibular angle. In group B unilateral curved ostectomy was performed with partial masseter resection. In group C unilateral angle splitting ostectomy, while in group D unilateral dissection of the masseter muscle was conducted. The constitution and its ratio of collagen fiber in the interface were observed at 1-month, 2-month, 3-month, and 6-month after operation. Results On the changes of collagen fiber in the process of muscular reattachment, at 1-month post-operation, the constitution of collagen fiber (types Ⅰ and Ⅲ) in groups A and B were significantly different from that of control group (P<0.05). However, both groups C and D had no statistic difference from control group (P>0.05). At 2-month, 3-month and 6-month post-operation, those of all experimental groups had no statistic difference from control group. And with time, the percentage of collagen fiber type Ⅰ increased and type Ⅲ decreased gradually. Conclusion The recovery sequences of masseter muscle reattachment in this study are firstly group C, secondly group A and finally group B. It suggests that the recoveries of mastication and other oral activities are different. Group B turns out to be with a slow muscle reattachment. Thus, we recommend treating different kinds of mandibular hypertrophy with different ostectomies.

Objective To investigate the changes of the masseter muscle following osteotomy of the prominent mandibular angle and to provide guidance for the resection of mandibular angle. Methods Ten goats were equally divided into two groups. In group A we performed unilateral curved osteotomy of the mandibular angle, and in group B we performed unilateral dissection of the masseter muscle. The cross section area (CSA) and the sarcomere length of masseter muscle were measured beore and after operation. Results (1) Cross section area (CSA) of masseter muscle fiber in curved ostectomy group decreased at 1,2, 3 and 6 months after operation in different extent. Comparing with the control group, the difference was statistically significant (P<0.01). CSA of masseter muscle fiber in dissection group decreased 1 month postoperatively, which had significantly statistic difference with control group (P<0.01). But, they had no significant difference with control group at 2, 3, and 6 months after operation (P>0.01). (2) Sarcomere length of masseter muscle in curved ostectomy group decreased in 1 week, 1 and 2 months after operation, which had significantly statistic difference with control group (P<0.01). At 3 months after operation, sarcomere length recovered to normal. In dissection group, sarcomere length decreased in 1 week and 1 month after operation, which had significantly statistic difference with control group (P<0.01). At 2 month after operation, it recovered to normal. Conclusion Certain extent of atrophy does happen to masseter muscle after mandibular angle ostectomy. Meanwhile, these changes do not significantly impair the function of masseter muscle. According to this, we suggest a simple mandibular angle ostectomy without partial resection of masseter muscle in case of mild to morderate mandibular angle hypertrophy. Doing so, we can not only achieve the cosmetic effect but also reduce the implications.

The gene expressions of codeinone reductase (COR) and berberine bridge enzyme (BBE) in Papaver somniferum were blocked by RNA hairpin of RNA interference (RNAi). The complete sequences of COR and BBE genes were cloned by reverse transcription-polymerase chain reaction (RT-PCR), the results of homology comparison revealed that the cloned COR and BBE genes had high homology with the other gene family members reported in the GenBank. The target sequences of COR and BBE genes were screened in accordance with the design principle of RNAi, a 643 bp fusion gene was obtained by the method of overlapping PCR, then plant expression vector ihpRNA was constructed based on intermediate vector pHANNIBAL and plant expression vector pCEPSPS. With that 78 transgenic plants were obtained through Agrobacterium-mediated and 17 positive plants were screened by PCR, that could initially indicate that the target fragments of COR and BBE gene had been integrated into tobacco genome.
Artificial Gene Fusion ; Genetic Vectors ; NAD (+) and NADP (+) Dependent Alcohol Oxidoreductases ; genetics ; Oxidoreductases, N-Demethylating ; genetics ; Papaver ; enzymology ; genetics ; Plants, Genetically Modified ; enzymology ; genetics ; RNA Interference ; RNA, Small Interfering ; Tobacco ; genetics ; Transformation, Genetic

Objective To analyze the clinical features and to explore the potential mode of genetic transmission of vitiligo vulgaris in Chinese Han population. Methods Information on the clinical features and the familial history of patients with vitiligo vulgaris was collected by a uniform questionnaire. The data were inputted into a data base, then analyzed by Epi Info 5.0 and SPSS 10.0 software packages. A complex segregation analysis was conducted using the REGTL program in SAGE 3.1 package in order to propose the putative mode of genetic transmission for vitiligo vulgaris. Results The mean age of onset of vitiligo vulgaris of the males and females was about 19 years old. In the severity of the disease and the season of onset, there was no significant difference between males and females. For the patients with vitiligo vulgaris, there was a female preponderance of complicating with autoimmune disorders, especially hyperthyroidism. The prevalence rates of vitiligo in patients′ relatives were significantly higher than that in general population. The mode of transmission of vitiligo vulgaris was one of the multifactorial inheritance according to the results obtained by the complex segregation analysis. Conclusions There are no differences in the prevalence and the age of onset between males and females patients. There is a marked familial aggregation in the patients with vitiligo vulgaris. The familial clustering might result from the polygenic additive genetic background and common environmental factors. It is suggested that vitiligo vulgaris be a multifactorial complex disease.

OBJECTIVETo detect and study the types of Borrelia burgdorferi sensu lato in ticks and rodents from Da Xing-An Mountains Forest areas of China.

METHODSNested PCR was performed to amplify 5S-23S rRNA intergenic spacer of B. burgdorferi. Positive products were analysed by restriction fragment length polymorphism (RFLP) and single strand conformation polymorphism (SSCP), specimens showing unique RFLP profile were sequenced and analysed.

RESULTS1336 Ixodes persulcatus, 144 Dermacento silvarum, 144 Haemaphysalis concinna and 145 rodents of 9 species were collected from 16 sections of Da Xing-An Mountains Forest areas of China. Specific fragments were amplified from 293 I. persulcatus and 6 D. silvarum and 5 rodents of 4 species. B. burgdorferi was not detected in H. concinna. Among the positively tested I. persulcatus, 209 contained B. garinii genospecies and 45 contained B.afzelii genospecies based on RFLP. Moreover, B.garinii genospecies consisted of B. garinii 20047 and B. garinii NT29. 17 adult I. persulcatus were simultaneously infected with B. garinii 20047 and B. garinii NT29. Nine adult I. persulcatus were simultaneously infected with B. garinii 20047 and B. afzelii. Four adult I. persulcatus were simultaneously infected with B. garinii 20047 and B. garinii NT29 and B. afzelii. Two D. silvarum were infected with B. garinii 20047, 1 D. silvarum with B. garinii 20047, 2 D. silvarum with B. afzelii. 3 rodents were infected with B. garinii 20047 while 2 rodents were infected with B. garinii NT29. Mixed infection was not found in D. silvarum and rodents. In addition, nine I. persulcatus and one D. silvarum specimens showed unique RFLP pattern. Data from sequential analysis showed that they all belonged to B. garinii. PCR-SSCP profiles of 5S-23S rRNA intergenic spacer of B. burgdorferi in the positive specimens exceeded 36 types; B. garinii 20047 showed 16 types while B. garinii NT29 showing 11 types, B. afzelii showing 9 types. SSCP profiles of the specimens coinfected with multiple B. burgdorferi was relatively complex.

CONCLUSIONThe infection of B. burgdorferi was found in the ticks and rodents in Da Xing-An Mountains Forests areas. The infection rate of I. persulcatus was high. B. garinii was predominant genospecies, and the population of B. burgdorferi was heterogeneous in the area. Mixed infections of different B. burgdorferi genospecies in ticks were found. I. persulcatus and Clethrionomys rufocanus were possibly served as major vector and major host for B. burgdorferi, respectively, suggesting that further study is needed to confirm the coinfection in humans and animals in this region.

Animals ; Borrelia burgdorferi Group ; genetics ; isolation & purification ; China ; epidemiology ; Humans ; Lyme Disease ; epidemiology ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single-Stranded Conformational ; RNA, Bacterial ; analysis ; Rodentia ; microbiology ; Ticks ; microbiology ; Trees

OBJECTIVETo study in the linkage between eotaxin-3 gene polymorphisms and allergic asthma susceptivity, blood plasma IgE or peripheral blood eosinophil in adult population of Han nationality from Hubei province of China.

METHODSPolymerase chain reaction (PCR), single strand conformation polymorphism (SSCP), tetra-primer PCR technique and restriction analysis were applied to identify the single nucleotide polymorphism.

RESULTSThe allele frequency of eotaxin-3 +2497 G, total levels of plasma IgE and peripheral blood eosinophil counts revealed the significant difference between control and allergic asthma group, that the P value was 0.011, 0.021 or 0.029 respectively. The allele frequency of eotaxin-3 +77 T and total levels of plasma IgE showed to have no significant difference between control and allergic asthma group, that the P value was 0.824 and 0.473 respectively. However, the peripheral blood eosinophil counts was significantly different between control and allergic asthma group, and the P value was 0.044.

CONCLUSIONSingle nucleotide polymorphism of eotaxin-3 +2497 is associated with the asthma susceptibility, peripheral eosinophil counts and total levels of plasma IgE in adult population from Hubei province, and polymorphism of +77 is associated with peripheral eosinophil counts.

Adult ; Alleles ; Asthma ; genetics ; immunology ; Base Sequence ; Chemokine CCL26 ; Chemokines, CC ; genetics ; China ; ethnology ; DNA Mutational Analysis ; Female ; Genetic Predisposition to Disease ; Humans ; Immunoglobulin E ; blood ; Male ; Middle Aged ; Molecular Sequence Data ; Polymorphism, Genetic ; Young Adult

Adult ; Female ; Humans ; Hydrothorax ; diagnosis ; etiology ; therapy ; Liver Cirrhosis ; complications ; Male ; Middle Aged ; Talc ; therapeutic use ; Thoracoscopy ; methods

OBJECTIVETo investigate the association between interferon regulatory factor 6 (IRF6) gene polymorphism and non-syndromic oral clefting (NSOC).

METHODSExperimental group consisted of 186 Ningxia NSOC patients, their parents (183 fathers and 174 mothers), 172 core families (patient+parents), and control group consisted of 200 normal children. DNA was extracted and PCR-restriction fragment length polymorphism (PCR-RFLP) was used to identify the genotypes of the samples, case-control analyses and transmission-disequilibrium test (TDT) were carried out.

RESULTSCompared with control group, there were significant differences in both rs642961's and rs4844880's AA genotype and A allele among NSOC patients (P < 0.05), but no difference in cleft palate (P = 0.15, P = 0.967, respectively). In TDT analysis, the A allele of rs642961 had a strong over-transmission in NSOC (P < 0.05), so did the rs4844880'A allele (P < 0.05), but neither of them had significant difference in cleft palate (P = 0.91, P = 0.95, respectively).

CONCLUSIONIRF6 gene polymorphism is associated with NSOC.

Case-Control Studies ; Cleft Palate ; genetics ; Humans ; Interferon Regulatory Factors ; genetics ; Polymorphism, Genetic

OBJECTIVETo evaluate the role of homozygosity mapping in the fine mapping of the genes responsible for the rare autosomal recessive diseases.

METHODSPolymerase chain reaction-single sequence length polymorphism was used to genotype the family members from 8 families with osteoporosis-pseudoglioma syndrome(OPS) for 14 polymorphic loci within candidate region. The OPS candidate region was narrowed by searching for homozygous region in affected.

RESULTSThe OPS candidate region was narrowed to a 1 cM interval between D11S1296 and D11S4136.

CONCLUSIONHomozygosity mapping is a powerful method for mapping and narrowing the candidate region of the genes responsible for the rare autosomal recessive diseases.

Abnormalities, Multiple ; genetics ; pathology ; Chromosome Mapping ; methods ; Chromosomes, Human, Pair 11 ; genetics ; Eye Diseases ; pathology ; Family Health ; Female ; Genetic Predisposition to Disease ; genetics ; Homozygote ; Humans ; Male ; Microsatellite Repeats ; Osteogenesis Imperfecta ; pathology ; Pedigree ; Syndrome