Objective To study the inhibitory effect of pterin acid (PTA) against ricin and recombinant ricin A chain protein. Methods Luciferase protein synthesis inhibition assay in a cell-free system and in vitro cytotoxicity experiments were performed to assess the biological activity of ricin and rRTA treated with PTA.Results The result showed that PTA could significantly inhibit the activity of ricin and rRTA in a dose-dependent manner.Conclusion PTA might be used as a small molecular probe to develop an evaluating system for ricin/RTA small molecular inhibitor in vitro. The cell-free system adopted in the current study could also serve as a necessary basis for screening some novel small molecular compounds against ricin and RTA in the future.

The genome of CK/CH/SD09/005, an isolate of infectious bronchitis virus (IBV), was characterized to enable the further understanding of the epidemiology and evolution of IBV in China. Twenty-five pairs of primers were designed to amplify the full-length genome of CK/CH/SD09/005. The nucleotide sequence of CK/CH/SD09/005 was compared with reference IBV strains retrieved from GenBank. The phylogenic relationship between CK/CH/SD09/005 and the reference strains was analyzed based on S1 gene sequences. The complete genome of CK/CH/SD09/005 consisted of 27691 nucleotides (nt), excluding the 5' cap and 3' poly A tail. The whole-genome of CK/CH/SD09/005 shared 97 - 99% nucleotide sequence homology with the GX-NN09032 strain, which was the only complete genome that was closely related to CK/CH/SD09/005. When compared with all reference strains except GX-NN09032, CK/CH/SD09/005 showed the highest similarity to ck/CH/LDL/091022 and SDIB821/2012 (QX-like) in the replicase gene (Gene 1) and 3'UTR, with a sequence identity rate of 97% and 98%, respectively. However, CK/CH/SD09/005 exhibited lower levels of similarity with ck/CH/LDL/091022 and SDIB821/2012 in S-3a-3b-3c/ E-M-5a-5b-N with a sequence identity of 72% - 90%. CK/CH/SD09/005 showed the highest level of nucleotide identity with Korean strain 1011, and Chinese strains CK/CH/LXJ/02I, DK/CH/HN/ZZ2004 and YX10, in ORF 3c/E (97%), 5a (96%), 5b (99%) and N (96%), respectively. ORFs 3a, 3b and M of CK/CH/SD09/005 exhibited no more than 90% homology with the reference strains, excluding GX-NN09032. The phylogenic analysis based on the S1 gene revealed that CK/CH/SD09/005 and 39 published strains were classified into seven clades (genotypes). CK/CH/SD09/005 was distributed in clade IV with several isolates collected between 2007 and 2012. CK/CH/SD09/005 showed 66% - 69% and 72% - 81% nucleotide identities with the IBV strains of other six clades in the S1 and S2 subunits, respectively. More over, multiple substitutions were found throughout the entire S gene of CK/CH/SD09/005, while insertions and deletions were located within the S1 gene. These results indicated that CK/CH/SD09/005 is a novel variant that may be derived from the QX-like strains that are prevalent in China. Multiple genetic mechanisms, including recombinations, mutations, insertions and deletions, are likely to have contributed to the emergence of this IBV strain.
Animals ; Chickens ; China ; Coronavirus Infections ; veterinary ; virology ; Genome, Viral ; Genomics ; Infectious bronchitis virus ; classification ; genetics ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Poultry Diseases ; virology ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics

Thirteen isolates of Class I Newcastle disease virus obtained from healthy poultry in China during 2008 were characterized genotypically in this study. All the isolates were proved to be lentogenic strains based on the deduced amino acid sequence of the Fusion protein gene. Molecular epidemiological analysis showed that 13 isolates could be subdivided into 2 distinct genotypes, 11 isolates belonged to genotype 2, and other 2 isolates belonged to genotype 3. Results indicated two genotypes of Class I Newcastle disease virus might widely exist in domestic poultry in China.
Animals ; Birds ; China ; epidemiology ; Genotype ; Humans ; Molecular Epidemiology ; methods ; Newcastle Disease ; epidemiology ; virology ; Newcastle disease virus ; classification ; genetics ; pathogenicity ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Fusion Proteins ; genetics

Bletilla striata has been used as traditional Chinese medicine for several centuries. In recent years, the quality and quantity of wild B. striata plants have declined sharply due to habitat deterioration and human over-exploitation. Therefore, it is of great urgency to evaluate and protect B. striata wild plant resource. In this study, sequence-related amplified polymorphism (SRAP) markers were applied to assess the level and pattern of genetic diversity in twelve populations of B. striata. The results showed a high level of genetic diversity (PPB = 90.48%, H = 0.349 4, I = 0.509 6) and moderate genetic differentiation among populations (G(st) = 0.260 9). Based on the unweighted pair-group method with arithmetic average (UPGMA), twelve populations gathered in three clusters. The cluster 1 included four populations. There are Nanjing, Zhenjiang, Xuancheng and Hangzhou. The seven populations which come from Hubei Province, Hunan Province, Jiangxi Province and Guizhou Province belonged to the cluster 2. The cluster 3 only contained Wenshan population. Moreover, Mantel test revealed significant positive correlation between genetic distances and geographic distances (r = 0.632 9; P < 0.000 1). According to the results, we proposed a series of conservation consideration for B. striata.
China ; Genetic Markers ; Genetic Variation ; Genetics, Population ; Orchidaceae ; genetics ; Phylogeny ; Plants, Medicinal ; genetics

Increased vascular endothelial growth factor (VEGF) biosynthesis in vascular endothelial cells has been reported to play an obligatory role in promoting angiogenesis. Nevertheless, the intracellular signaling mechanisms of hypoxia-induced VEGF release remain largely unknown. Human umbilical vein endothelial cell lines (ECV304) were cultured in normoxic or hypoxic conditions for 12 approximately 24 h and harvested for determination of VEGF mRNA expression and phosphorylation of ERK1/2 and p38 mitogen-activated protein kinase (p38 MAPK) by real-time reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. Secreted VEGF protein was measured by enzyme-linked immunosorbent assay (ELISA). It has reported that PD98059, an ERK inhibitor, was able to blunt the hypoxia-induced activation of the expression of VEGF gene. In accordance with this report, an increase in ERK1/2 phosphorylation and VEGF biosynthesis was observed in ECV304 cells cultured in hypoxia, and this increase was blocked by PD98059. The novel finding of the present study is that an activation of p38 MAPK is involved in hypoxia-induced increase in VEGF biosynthesis. SB202190, an inhibitor of p38 MAPK was able to blunt the hypoxia-induced increase in VEGF biosynthesis. These dada provide the first direct evidence for a role of p38 MAPK in mediating hypoxia-induced increase in VEGF biosynthesis in human endothelial cells.
Cell Hypoxia ; Cell Line ; Endothelial Cells ; cytology ; metabolism ; Humans ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Phosphorylation ; RNA, Messenger ; genetics ; metabolism ; Umbilical Veins ; cytology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism ; physiology

OBJECTIVETo observe the effect of early treatment with acupuncture and motortherapy on developmental quotient (DQ) of cerebral palsy high risk infants.

METHODSSixty cerebral palsy high risk infants were divided into an acupuncture combined with motortherapy group (treatment group) and a control group, 30 cases in each group. Changes of DQ were investigated by the children mental development scale.

RESULTSThe DQ in the treatment group was significantly higher than that in the control group (P < 0.001), with a very significant difference between the two groups in the different grades of DQ (P < 0.005). Incidence of cerebral palsy in the treatment group significantly lower than that in the control group (P < 0.005).

CONCLUSIONAcupuncture combined with motortherapy can effectively improve intelligence level and motor function, and reduce the incidence of cerebral palsy for cerebral palsy high risk infants at early stage.

Acupuncture Therapy ; Cerebral Palsy ; psychology ; therapy ; Child Development ; Exercise Therapy ; Female ; Humans ; Infant ; Infant, Newborn ; Male

OBJECTIVETo investigate the effect of the third-party bone marrow-derived mesenchymal stem cells (BMSCs) on the allogeneic skin transplantation.

METHODS40 female C57BL/6 mice and 50 male BALB/C mice were respectively used as donors and recipients of skin transplantation. 50 BALB/C mice were divided randomly into 5 groups: Blank control group, Cyclophosphamide group BMSCs group, Cyclophosphamide + BMSCs group and CM-DiI staining group, with 10 mice in each group. Before skin transplantation, high-dose abdominal injection of Cyclophosphamide (200 mg/kg, 2 d) was performed in recipient mice. On the transplantation day, a bonus of 1 x 10(5) BMSCs of the SD rat (SD-BMSCs) were injected through the tail vein. The observation of skin grafts, mixed lymphocyte culture (MLC), HE staining, the observation of CM-DiI-labeled SD-BMSCs and FACS were used.

RESULTSThe skin graft survival time was significantly prolonged in the Cyclophosphamide + BMSCs group, as compared with the blank control group, the Cyclophosphamide group, the BMSCs group respectively. When BMSC and lymphocyte mixed at the ratio of 1:1 and 1:10, rat BMSCs inhibited T lymphocyte proliferation. More angiogenesis and less lymphocyte infiltration were found in the experimental group than them in other groups. Red fluorescent cells were found in CM-DiI staining group under long-term observation. The SD-BMSCs can he detected by flow cytometry in the cell group and the Cyclophosphamide + BMSCs group.

CONCLUSIONSBMSCs can survive in the heterogeneous recipient body; the third-party BMSCs transplantation can prolong skin graft survival time; BMSCs can inhibit T lymphocyte activation and proliferation.

Animals ; Cells, Cultured ; Female ; Male ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; Transplantation, Homologous

Objective: To evaluate the clinical efficacy of the modified Stoppa approach in the surgical treatment of pelvic fractures of Tile type C combined with acetabular both-column fractures. Methods: Between April 2014 and April 2017, a total of 14 patients were treated by open reduction and internal fixation (ORIF) through the modified Stoppa approach at Department of Orthopaedics, Foshan Gaoming Hospital for pelvic fractures of Tile type C combined with acetabular both-column fractures. They were 10 men and 4 women, with an average age of 36.4 years (from 23 to 57 years). The modified Stoppa approach was used exclusively in 4 cases, in combination with the iliac fossa approach in 3 ones, in combination with the Kocher-Langenbeck approach in 4 ones and in combination with the Kocher-Langenbeck and iliac fossa approaches in 3 ones. In all the patients, the pelvic reconstructive plate and screws and lag screws were used. The operation time, intraoperative bleeding volume, postoperative fracture reduction, fracture union time, efficacy and complications were recorded. Results: The operation time averaged 170 min (from 110 to 330 min) and the intraoperative bleeding 420 mL (from 240 to 1 100 mL). There were no operative complications. By the Matta evaluation, the postoperative reduction was rated as excellent in the 14 pelvic fractures and 9 acetabular both-column fractures and as good in 5 acetabular both-column fractures. Of this series, 13 patients were followed up for an average of 19 months (from 12 to 29 months) and one was lost to the follow-up. The fractures got united after an average time of 3.4 months (from 2.7 to 4.6 months). Screw loosening was observed in one case and mild limitation to hip flexion in one. Follow-ups found no lateral ventral syndrome or femoral head necrosis. Their Harris hip scores at the last follow-up ranged from 70 to 94 points, averaging 84 points. The function of the affected hip was excellent in 6 cases, good in 5 and fair in 2. Conclusion The modified Stoppa approach may be used exclusively or in combination with other approaches to treat effectively the pelvic fractures of Tile type C combined with acetabular both-column fractures, leading to good short-term clinical outcomes.

Objective To construct the protein-protein interaction network of Parkinson's disease (PD) associated proteins. Methods PD-related proteins and their interaction proteins were collected by bioinformatics method from HGNC, OPHID and UniHI database. A protein-protein interaction network of PD associated proteins was designed by ProteoLens software; each protein was evaluated by its contribution to the network and the relevance scores were calculated, thus the coefficient of association of each protein in the network was noted. Results A protein-protein interaction network containing 463 PD associated proteins and 767 their interaction proteins was obtained with its index aggregation reaching 90.5% (P=0.008). The relevance scores of SNCA, PARK2, DRD2, HTRA2,NDUFV2, DJ1, DRD1, DRD3, TRAP1 and ND3 were much higher than that of the others, which indicated their important roles in the pathogenesis of PD. The relevance scores of APP, UBE2I, CLIC6 and UBB were a little higher among all the preteins, indicating that they also participated in the pathogenesis of PD. Some novel proteins, such as FLNA, FREQ, BIRC7, EPB41, EPB41L1, GIPC1,GNAZ, GRB2, KCNJ9, MAPK1, BAG5 and CYC, may also involved in the pathogenesis of PD.Conclusion A scientific and practical protein-protein interaction network of PD associated proteins is successfully constructed, which further confirms the role of some proteins in the pathogenesis of PD.Some novel proteins that might involve in PD are obtained too.

OBJECTIVETo study the clinical and genetic characteristics of a Chinese family with benign familial convulsions (BFNC).

METHODSThe clinical data of this family was analyzed. The blood samples were collected from 13 members of this family. By four microsatellite markers which are located in the gene loci of both K+ channel KCNQ2 and KCNQ3, the linkage analysis was performed in the family. With DNA direct sequencing and restriction endonuclease cutting analysis, the mutation analysis of KCNQ3 gene was made for the proband, other 12 family members and 76 unrelated normal individuals.

RESULTSThere were 7 patients with BFNC observed in the three generation of family. The BFNC seizures of all patients disappeared during one month and no recurrence of seizures was found. The linkage analysis suggested the disease gene linked to KCNQ3 gene locus in the family. The mutation 988(C to T) of KCNQ3 gene was found in the proband by DNA-direct sequencing. Cosegregation of this mutation with BFNC was confirmed by restriction endonuclease cutting analysis.

CONCLUSIONChinese patients with BFNC can be caused by KCNQ3 gene mutation.

Base Sequence ; Child ; China ; DNA Mutational Analysis ; Epilepsy, Benign Neonatal ; genetics ; pathology ; Family Health ; Female ; Genetic Linkage ; genetics ; Genotype ; Humans ; KCNQ3 Potassium Channel ; genetics ; Male ; Mutation ; Pedigree ; Sequence Analysis, DNA