Objective To construct the eukaryotic expression vector pEGFP-N1/IL-37b and analyze the expression of IL-37 gene in THP-1 cells. Methods Total RNA was extracted from human peripheral blood mononuclear cells ( PB-MCs) and the coding region of IL-37b gene was amplified by RT-qPCR. Then, the gene was cloned into pEGFP-N1 eu-karyotic expression vector. After transfected the recombinant plasmid into THP-1 cells, the expression of IL-37 was detec-ted by RT-qPCR and Western blot. Results Double restriction enzyme digestion and gene sequencing showed that IL-37b gene was correctly inserted into the eukaryotic expression vector pEGFP-N1. RT-qPCR and Western blot showed that the IL-37 expression level was increased significantly (P<0. 01) after transfection in THP-1 cells. Conclusions We successful-ly constructed a novel anti-inflammatory cytokine IL-37 eukaryotic expression vector pEGFP-N1/IL-37b, which lays a foun-dation for further study on IL-37 functions and its association with related diseases.

PEI Xue-yi has 60 years,experience in treating pediatric diseases with traditional Chinese medicine,especially obtains good effect in children's nephrapiathy.Based on the regulation and characteristics of children's zang-fu,yin-yang,blood-qi and children's nephrapiathy,he makes different therapeutic plans in different phases.In clinical treatment,he emphasizes the lung,the spleen and the kidney and combines syndrome differentiation with disease diagnosis.Eliminaing pathogens,excessive damp-heat is used in acute phase and strengthening healthy qi,securing the spleen and kidney is used in recovery phase.Children's nephrapiathy is treated by three phases as the edema,protein urine and rescovery stage.He adopts the methods of dispersing lung qi and invigorating spleen for diuresis,clearing heat-damp,nourishing spleen and kidney,nourishing yin for protecting lower jiao.

Cardiac toxicity is found in frequently used chemotherapeutic agents.There are many factors related to the cardiac toxicity caused by chemotherapeutic agents.The common cardiovascular complications include heart failure,ischemia,hypertension,hypotension,edema,QT prolongation,pleural effusion,pericardial effusion,bradyarrhythmia and thromboembolism.It is necessary to monitor the left ventricular function before and after chemotherapy and take effective measures to protect myocardium.

In this paper, the chloroplast psbK-psbI intergenic spacers of 18 species of Dendrobium and their adulterants were amplified and sequenced, and then the sequence characteristics were analyzed. The sequence lengths of chloroplast psbK-psbI regions of Dendrobium ranged from 474 to 513 bp and the GC contents were 25.4%-27.6%. The variable sites were 71 while the informative sites were 46. The inter-specific genetic distances calculated by Kimura 2-parameter (K2P) of Dendrobium were 0.006 1-0.058 1, with an average of 0.028 4. The K2P genetic distances between Dendrobium species and Bulbophyllum odoratissimum were 0.093 2-0.120 4. The NJ tree showed that the Dendrobium species can be easily differentiated from each other and 6 samples of the inspected Dendrobium species were identified successfully through sequencing the psbK-psbI intergenic spacer. Therefore, the chloroplast psbK-psbI intergenic spacer can be used as a candidate marker to identify Dendrobium species and its adulterants.
Chloroplasts ; DNA, Chloroplast ; genetics ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Dendrobium ; classification ; genetics ; Plants, Medicinal ; classification ; genetics

In order to identify Peucedani Radix, Peucedani Decursivi Radix and their adulterants, the internal transcribed spacer 2 (ITS2) regions of Peucedani Radix, Peucedani Decursivi Radix and their adulterants were amplified and bidirectionally sequenced based on the Principles for Molecular Identification of Traditional Chinese Materia Medica Using DNA Barcoding, which has been promulgated by Chinese Pharmacopoeia Commission. Sequences were analyzed and assembled by Codon Code Aligner V3. 7.1. The relevant data were analyzed by MEGA 5. 0. Species identification analyses were performed by using the nearest distance methods and neighbor-joining (NJ) methods. The result showed that the ITS2 sequence lengths of Peucedani Radix were 229-230 bp and the average intra-specific genetic distances were 0.005. The ITS2 sequence lengths of Peucedani Decursivi Radix were 227 bp and the sequences contained no variation site. The average inter-specific K2P genetic distance of Peucedani Radix, Peucedani Decursivi Radix and their adulterants species were 0.044 and 0.065 respectively. The minimum inter-specific divergence is larger than the maximum intra-specific divergence of Peucedani Decursivi Radix. The nearest distance methods and NJ trees results indicated that Peucedani Radix, Peucedani Decursivi Radix and their adulterants species could be identification clearly. The ITS2 regions can stably and accurately distinguish Peucedani Radix, Peucedani Decursivi Radix and their adulterants.
Apiaceae ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Ribosomal Spacer ; Drug Contamination

OBJECTIVETo identify the Cynomorii Herba and its analogues species using DNA barcoding technique.

METHODTotal genomic DNA extracted from all materials using the DNA extraction kit. The internal transcribed spacer 2 (ITS2) regions were amplified using polymerase chain reaction (PCR), and purified PCR products were sequenced bi-directionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner 3.7.1. The Kimura 2-Parameter (K2P) distances and GC content were computed using MEGA 5. 0. Species identification analyses were conducted through the species identification system for traditional Chinese medicine and neighbor-joining (NJ) trees.

RESULTThe ITS2 sequence lengths of Cynomorii Herba were 229 bp. The average intra-specific genetic distances of Cynomorii Herba were 0.003. The average inter-specific genetic distances between Cynomorii Herba and its adulterants species were 0.760. The results showed that the minimum inter-specific divergence is larger than the maximum intra-specific divergence. The species identification system for traditional Chinese medicine and NJ trees results indicated that Cynomorii Herba and its adulterants species can be easily identification.

CONCLUSIONThe ITS2 region is an efficient barcode for identification of Cynomorii Herba, which provide a new technique to ensure clinical safety in utilization of traditional Chinese medicine.

Cynomorium ; classification ; genetics ; DNA Barcoding, Taxonomic ; DNA, Intergenic ; genetics ; DNA, Plant ; genetics ; Polymerase Chain Reaction

Acute Disease ; Adolescent ; Arthralgia ; diagnosis ; Child ; Child, Preschool ; Diagnostic Errors ; Female ; Humans ; Leukemia ; diagnosis ; Male

OBJECTIVETo investigate the clinical effect of atlantoaxial pedicle screw internal fixation for treatment of atlantoaxial dislocation.

METHODSSixteen patients with atlantoaxial dislocation were treated from Dec. 2005 to June 2007, included 10 males and 6 females, aged from 38 to 45 years old (means 40.5 years). Among them 12 patients combined with nerve injury, according to ASIA grade: there were 3 cases in grade B, 5 cases in grade C, 4 cases in grade D. All patients received preoperative CT, radiograph and skull traction. Intraoperative posterior approach general spine pedicle screw-rod orthopaedics fixation system used and iliac bone block were implanted in space of posterior atlantal arch and axial vertebral plate. The outcome and complications were observed in the near future.

RESULTSThere was no vascular or neural injury found. The patients were followed up for 12 to 24 months (means 18 months). All head pain, acid storm symptoms were improved after operation. According to the Odom's clinical efficacy evaluation standard, 12 cases were excellent, 4 were good. Eleven cases of 12 with nerve injury recovered significantly. By ASIA classification: 1 cases was in grade B, 2 cases were in grade C, 5 cases were in grade D, 4 cases were in grade E. No looseness or breakage of screw occurred. Bony fusion was achieved in all cases.

CONCLUSIONPosterior approach atlantoaxial pedicle screw internal fixation have the advantages of direct screw placement, short-segment fusion, intraoperative reduction, fixation reliable, high fusion rate, and it can restablish the upper cervical vertebrae stability and help to recover the spinal cord and nerve function.

Adult ; Atlanto-Axial Joint ; diagnostic imaging ; injuries ; physiopathology ; surgery ; Bone Screws ; Female ; Fracture Fixation, Internal ; instrumentation ; Humans ; Joint Dislocations ; diagnostic imaging ; physiopathology ; surgery ; therapy ; Male ; Middle Aged ; Tomography, X-Ray Computed ; Treatment Outcome

Cell Line, Tumor ; DNA Repair ; DNA Repair Enzymes ; Hep G2 Cells ; Humans

BACKGROUND: All-trans retinoic acid (ATRA) promotes the osteogenic differentiation induced by bone morphogenetic protein 9 (BMP9), but the intrinsic relationship between BMP9 and ATRA keeps unknown. Herein, we investigated the effect of Cyp26b1, a critical enzyme of ATRA degradation, on the BMP9-induced osteogenic differentiation in mesenchymal stem cells (MSCs), and unveiled possible mechanism through which BMP9 regulates the expression of Cyp26b1. METHODS: ATRA content was detected with ELISA and HPLC–MS/MS. PCR, Western blot, and histochemical staining were used to assay the osteogenic markers. Fetal limbs culture, cranial defect repair model, and micro–computed tomographic were used to evaluate the quality of bone formation. IP and ChIP assay were used to explore possible mechanism. RESULTS: We found that the protein level of Cyp26b1 was increased with age, whereas the ATRA content decreased. The osteogenic markers induced by BMP9 were increased by inhibiting or silencing Cyp26b1 but reduced by exogenous Cyp26b1. The BMP9-induced bone formation was enhanced by inhibiting Cyp26b1. The cranial defect repair was promoted by BMP9, which was strengthened by silencing Cyp26b1 and reduced by exogenous Cyp26b1. Mechanically, Cyp26b1 was reduced by BMP9, which was enhanced by activating Wnt/b-catenin, and reduced by inhibiting this pathway. b-catenin interacts with Smad1/5/9, and both were recruited at the promoter of Cyp26b1. CONCLUSIONS Our findings suggested the BMP9-induced osteoblastic differentiation was mediated by activating retinoic acid signalling, viadown-regulating Cyp26b1. Meanwhile, Cyp26b1 may be a novel potential therapeutic target for the treatment of bone-related diseases or accelerating bone-tissue engineering.