A rare strain of actinomycetes, with broad-spectrum antimicrobial activity, was isolated from the soil samples from the farmland in the area of Yaohu lake in Nanchang. The information about the taxonomic identification, such as the morphology, physiological properties, cell components and 16S rRNA gene se-quences, suggested that the rare strain of actinomycetes was identified as Micromonospora carbonacea.

Based on the strain of Micromonospora carbonacea JXNU-1 with board-spectrum antimicrobial activity, the technology for the isolation and purification of antibiotic from the fermentation broth of the Micromonospora carbonacea, and its physical-chemical properties were studied. The results showed that, the antibiotic was stable under the condition of high temperature and alkali, but not in acid solution. After the pretreatment of centrifugation and filtration to remove the cells and lipids, the antibiotic was absorbed to negative exchange resin, and the impurity was excluded when 2 mol/L NaCl was used as primary eluent. The antibiotic could be eluted with 20% alcohol as eluent, and the eluting speed of the antibiotic was greatly accelerated as 2 mol/L NaCl was added into 20% alcohol as final eluent. Aqueous solution of the antibiotic was yielded from the alcohol-salt eluant by decompression concentration to wipe off alcohol and by dialysis to exclude salt. One active component was detected in antibiotic solution by paper chromatography, and theHPLC purity was over 99%. As the antibiotic shows positive color-forming reaction to Molish reagents, Benedict’s reagents and Diohenvlamine reagents, combined with the characteristics of absorption spectra, it is deduced that the antibiotic belongs to nucleoside antibiotics.

OBJECTIVETo understand if the Neuraminidase (N1) of Influenza A virus at the surface of yeast-displaying system, eukaryotic expression system and the infected cells could be used for anti-NA Abs screening, their activities and bindings to five candidate Abs were assayed.

METHODSThe surface NA expression was obtained by transfecting by recombinant NA constructors with specific tag-labels or live virus infection. The functional activity was measured by the fluorescent assay. Their bindings to the Abs were detected by flow cytometry.

RESULTSThe surface NAs presenting on the yeast-displaying system and eukaryotic expression system exhibited functional NA activities as the NA at the surface of virus-infected cells which showed affinities to Ab1, 4, and 5. The same bindings to Abl and 5 were found in the surface NA expressed by eukaryotic expression system while minor binding was observed in the yeast displayed-NA.

CONCLUSIONThe epitopes of yeast-displayed NA may be different from the NAs present at eukaryotic expression system and the infected cells which more likely suitable for the screening of anti-NA Abs.

Antibodies ; immunology ; Antigens, Surface ; genetics ; immunology ; Cell Line ; HEK293 Cells ; Humans ; Influenza A virus ; genetics ; immunology ; Neuraminidase ; genetics ; immunology ; Protein Binding ; Recombinant Proteins ; genetics ; immunology

Aged ; Cell Differentiation ; Humans ; Male ; Neoplasm Metastasis ; Neurosecretory Systems ; pathology ; Prostate-Specific Antigen ; blood ; Prostatic Neoplasms ; blood ; pathology

OBJECTIVETo explore the expression of origin recognition complex 1 (ORC1) during the DNA replication of vascular muscle cells (VSMC).

METHODSVSMC of thoracic aorta in rats were obtained by the adherence method of tissue culture. The cell synchrony was obtained by the method of double-thymidine block, colchicine treatment and serum starvation. The expression of ORC1 mRNA at different cell cycles of VSMC was determined by RT-PCR and the protein expression of ORC1 was analyzed by Western blot.

RESULTSCultured VSMC were identified by light microscope and immunocytochemistry. Significant expression of ORC1 mRNA and protein in a quiescent stage of VSMC were not observed. Upon synchronization, the expression of ORC1 mRNA was significantly higher at G(1)/S phase of VSMC than that at S and G(2)/M phases. The expression of ORC1 protein followed same changes as the ORC1 mRNA expression at different stages of cell cycles.

CONCLUSIONORC1 may be an important regulatory factor at the initiation of proliferative process of VSMC.

Animals ; Aorta, Thoracic ; cytology ; Blotting, Western ; Cell Cycle ; Cell Proliferation ; Cells, Cultured ; DNA ; genetics ; DNA Replication ; Gene Expression Regulation, Neoplastic ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; metabolism ; Origin Recognition Complex ; biosynthesis ; genetics ; physiology ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo summarize and analyze the diagnosis and arthroscopic treatment of osteochondral lesion of talus (OLT).

METHODSFrom 2000 to 2005 the data of 34 patients of OLT of the talus were retrospectively studied, including the symptom, physical examination, image, arthroscopic treatment All patients took X-ray and MRI examination before the arthroscopic surgery. Arthroscopic debridement was performed for all patients, in addition to drilling in 5 cases, and microfracture in 18 cases. Before operation, ankle-hindfoot score of American Orthopaedic Foot and Ankle Society (AOFAS) was 71 +/- 8, and the score of pain (visual analogue scale, VAS) was 7.5 +/- 1.3.

RESULTSWeight-bearing pain of the ankle joint aggravated after exercise was the predominant complaint of OLT. X-ray examination was negative in 13 cases, and all lesions were detected by MRI, which was significantly better than X-ray (chi2 = 16.07, P < 0. 001). Thirty-one patients were followed up for an average of 28 months. The average post-operative AOFAS was 91 +/- 9 (t = 9.147, P < 0.001); And VAS was 2.4 +/- 2. 3, which was significantly lower than that in pre-operation (t = 10.853, P < 0.001). Of the 31 patients, 27 (87.1%) had good or excellent results.

CONCLUSIONSMRI could improve the accuracy of diagnosis. The results of arthroscopic treatment for OLT are satisfactory.

Adolescent ; Adult ; Ankle Injuries ; diagnosis ; surgery ; Arthroscopy ; methods ; Cartilage, Articular ; injuries ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Retrospective Studies ; Talus ; injuries ; Treatment Outcome

OBJECTIVETo investigate regulation function of anodonta glucan HBP-A on chondrocytes through Wnt pathway in vitro.

METHODSRat chondrocytes were cultured and differentiated induced with IL-1beta (10 ng/ml) in vitro. Chondrocytes were divided into five groups:IL-13 group,IL-1beta + IWP-2 (5 microM,Wnt pathway inhibitor) group, IL-1beta + HBP-A (0.3 mg/ml) group and IL-1beta + IWP-2 + HBP-A group. Wnt-3a, beta-catenin (24 h,48 h,72 h) and MMP-13(72 h) genes expression were detected by Rt-PCR, while beta-catenin, MMP-13, Sox-9 and coll-II (48 h) protein expression were measured by Western-blot.

RESULTSAfter induction of IL-1beta, gene expression of Wnt-3a, beta-catenin and MMP-13 were increased,so were the protein expression of beta-catenin and MMP-13. In contrast,protein expression of Sox-9 and Coll-II were declined. Following addition of HBP-A, Wnt-3a, beta-catenin and MMP-13 were shown as induction of IL-1beta, but protein expression of Sox-9 and Coll-II were upgraded. Combining HBP-A with IWP-2 led to the lowest level in Wnt-3a, beta-catenin gene and beta-catenin protein expression and highest expression of Sox-9 protein.

CONCLUSIONHBP-A could not only delay the differentiation of chondrocytes through downgrading the signal expression of Wnt/beta-catenin,but also adjust the expression of Wnt-3a, beta-catenin and Sox-9 when combinated with the Wnt inhibitor.

Animals ; Anodonta ; chemistry ; Cell Differentiation ; drug effects ; Cells, Cultured ; Chondrocytes ; cytology ; drug effects ; metabolism ; Glucans ; pharmacology ; Interleukin-1beta ; metabolism ; Rats ; Wnt Signaling Pathway ; drug effects ; Wnt3A Protein ; genetics ; metabolism ; beta Catenin ; metabolism

OBJECTIVETo determine the genetic relationship of three species of official Rheum in molecular level.

METHODTwelve samples from three species of official Rheum were employed to be analyzed by the approach of sequence-related amplified polymorphism (SRAP). Systematic relationships were constructed based on the UPGMA method by TREECONW software.

RESULTA total of 272 bands were scored and 199 bands of them were polymorphic, which were up to 73.2% polymorphic ratio. Genetic similarity coefficient was changed from 0.578 4 to 0.941 6. The results indicated that there was abundant genetic diversity among the tested materials. The clustering analysis revealed that the results between SRAP marker and the traditional morphological characteristics was almost the same.

CONCLUSIONSRAP marker is suitable for variety identification and genetic relationship research in official Rheum.

Cluster Analysis ; Genetic Variation ; genetics ; Phylogeny ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Rheum ; classification ; genetics

BACKGROUNDCompared with traditional arthrotomy procedures, arthroscopic treatment for osteochondral lesions of the talus has some advantages. However, there has been considerable debate about the outcome predictors for this surgical technique. This study aimed to investigate the outcomes of arthroscopic treatment for osteochondral lesions of the talus, and analyze its outcome predictors.

METHODSClinical data of 48 patients with osteochondral lesions of the talus who underwent ankle arthroscopy were studied. Arthroscopic debridement was performed on all patients, and microfracture was also performed in 36 cases. Scores on a subjective satisfaction questionnaire, visual analog scale (VAS) for pain, and the American Orthopedic Foot & Ankle Society (AOFAS) ankle and hindfoot scores were obtained before and after surgery.

RESULTSFive patients lost to follow up. The other forty-three patients, 8 of whom were athletes, were followed up for an average of 23.9 months. The average AOFAS post-operative score was 90.16 +/- 9.96, compared with 70.81 +/- 6.96 before surgery (t = 9.353, P < 0.001). The VAS pain score after the operation (2.51 +/- 2.45) was significantly lower than that before the operation (6.95 +/- 1.40) (t = 8.647, P < 0.001). Of the 43 patients, 35 (81.4%) had good or excellent results. There was no significant difference in outcome between the medial and lateral groups (z = 0.205, P = 0.838), while a better outcome was found with lesions smaller than 10 mm than those with larger lesions (z = 2.199, P = 0.028). Age, sex, athletic profession and location of the lesion did not significantly correlate with outcomes.

CONCLUSIONSArthroscopic treatment is effective and safe for osteochondral lesions of the talus. A strong correlation was found between the size of the lesion and successful outcome.

Adolescent ; Adult ; Arthroscopy ; methods ; Female ; Humans ; Male ; Middle Aged ; Osteochondritis ; surgery ; Talus ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVE: To explore the relationship between the time-dependent level changes of microRNA and 18S rRNA and the different postmortem interval (PMI) in rat cardiac muscle. METHODS: SD rats were sacrificed by cervical dislocation and placed at ambient temperature 25 degrees C with a humidity of 50%. Total RNA was extracted from the rat cardiac muscle at different time points after death. The levels of miR-1-2 and 18S rRNA were examined using real-time PCR in rat cardiac muscle. The results were expressed by cycle threshold (Ct) value to explore relationship between PMI and Ct value, and the regression functions were established to estimate PMI. RESULTS: The miR-1-2 level in rat myocardial tissue showed no significant changes within 120 h after death, and then began to decline. The 18S rRNA level increased gradually within 96 h after death, and then declined slowly. The nonlinear relationships were established between Ct value (18S rRNA), deltaCt value (difference between 18S rRNA and miR-1-2) and PMI. The R2 of conics fitting were 0.9487 and 0.8072, respectively. CONCLUSION Ct value of 18S rRNA and deltaCt value present a good correlation with PMI, and can be markers for estimating early PMI.
Animals ; Forensic Pathology ; Male ; MicroRNAs/metabolism* ; Myocardium/pathology* ; Postmortem Changes ; RNA Stability ; RNA, Messenger/metabolism* ; RNA, Ribosomal, 18S/metabolism* ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction/methods* ; Time Factors