Objective To develop a method to prepare human articular cartilage derived microcarrier for both rapid propagating chondrocytes and being used as scaffold to support chondrogenesis. Methods Human articular cartilage was crushed into small pieces by muller after lyophilization, and sorted through two different meshes to collect only those specimens measuring 150-200 microns. Then, in turn, the specimens were subjected to 0.25% trypsin at 37 ℃ for 24 hours and 1% Triton X-100 for 72 hours, respectively. The specimens were observed by inverted phase contrast microscopy, and assessed by staining with haematoxylin-eosin, safranin-O (for GAG), as well as by the immunohistochemistry of aggrecan, collagen type Ⅱ. The microcarriers were seeded with human chondrocytes after being irradiated by 60Co. Results Using inverted phasecontrast microscope, the freezing-dry cartilage particles were observed as yellow, different shapes, and their surfaces were uneven, and with many pits. After treating with trypsin and Triton X-100, the microcarriers showed light yellow, without cartilage morphology. The microcarriers became flocculous or like a hairbrush, and the area of contacting surface significant increased. After culture with cartilage cell for 2 hours, lots of spherical chondrocytes adhered to the microcarriers. HE stain of section confirmed that the celluar constituents of the specimens were removed, the specimens stained weakly positive for GAG, negatively for aggrecan, and positively for collagen type Ⅱ, respectively. Conclusion The detergent and trypsin can remove the cellular constituents and knock out the aggrecan from human articular cartilage while maintaining collagen type Ⅱ and GAG, and made the cartilage pieces flocculous or hairbrush-like. The chondrocytes can be well maintained in human articular cartilage derived microcarriers. Human articular cartilage derived microcarriers were prepared successfullly.

ObjectiveTo evaluate the osteoinductivity of cationic liposome mediated recombined human bone morphogenetic protein-2 gene (rhBMP-2) transfer in skeletal muscle stem cells (SMSCs) of rabbits. MethodsSMSCs were transfered by pAGFP rhBMP-2 liposome complex. Gene transferred SMSCs were evaluated with immunohistochemistry.ResultsThe transfection rate of SMSCs transferred by cationic liposome mediated pAGFP-rhBMP-2 was 14.18%. Positive stain of rhBMP-2, alkaline phosphatase(ALP),collegenⅠhad been found in the transferred SMSCs.ConclusionQuantity of SMSCs is enough to be target cells of gene therapy. Cationic liposome can mediated bone morphogenetic protein-2 gene transfected SMSCs in vitro. Compared with AdrhBMP-2,the transfection rate of liposome mediated bone morphogenetic protein-2 gene transfected SMSCs is lower.

ObjectiveTo observe the effect of rabbit skeletal muscle stem cells (RSMSCs) modified by adenovirus mediated bone morphogenetic protein-2 (BMP-2) gene ex vivo in combination with demineralized bone matrix (DBM) on repair of longer bone defect in rabbit.MethodsModel of radial bone defects (20 mm) of rabbits was established. 50 rabbits were divided into 5 groups, group A (AdrhBMP-2 trusduced RSMSCs/DBM group), group B (adGFP trusduced RSMSCs/DBM group), group C (not trusduced RSMSCs/DBM group), group D (DBM group), and group E (untreated group). Roentgenographic, histologic, biomechanical, bone density of all animals were examined at the end of 4th and 6th week after surgery.ResultsAt 4th week, radial bone defects healed in group A. The healing rates from group A to group E were 100%, 50%, 33%, 0%, and 0% respectively at 6th week.ConclusionRSMSCs modified by AdrhBMP-2 ex vivo in combination with DBM can repair radial longer segemental defect in rabbit. It's possible to be used in the clinical treatment of longer segemental bone defect.

OBJECTIVE: A case of half sibling was determined with multiple genetic markers, which could be potentially applied for determination of half sibling relationship from same father. METHODS: Half sibling relationship was detected by 39 autosomal STR genetic markers, 23 Y-chromosomal STR genetic markers and 12 X -chromosomal STR genetic markers among ZHAO -1, ZHAO -2, ZHAO -3, ZHAO -4, and ZHAO-5. RESULTS: According to autosomal STR, Y-STR and X-STR genotyping results, it was determined that ZHAO-4 (alleged half sibling) was unrelated with ZHAO-1 and ZHAO-2; however, ZHAO-3 (alleged half sibling), ZHAO-5 (alleged half sibling) shared same genetic profile with ZHAO-1, and ZHAO-2 from same father. CONCLUSION It is reliable to use multiple genetic markers and family gene reconstruction to determine half sibling relationship from same father, but it is difficult to determination by calculating half sibling index with ITO and discriminant functions.
Alleles ; Chromosomes, Human, Y/genetics* ; Discriminant Analysis ; Gene Frequency ; Genetic Loci/genetics* ; Genetic Markers ; Genotype ; Humans ; Paternity ; Siblings

Objective To elucidate the prevalence and clinical characteristics of human metapneumovirus(hMPV)in hospital- ized children with respiratory infection.Methods A total of 452 hospitalized children with lower respiratory tract infection were observed from Aug 2004 to Jan 2005.Respiratory tract aspirates were collected from all patients within 48 hours after admis sion.The specimens were routinely tested for respiratory syncytial virus,influenza virus A and B,parainfluenza virus 1 to 3 and adenovirus by direct fluorescent assay(DFA).The 245 specimens negative by DFA were tested for hMPV by RT-PCR. PCR products of hMPV M gene from some patients were randomly selected for sequencing analysis.Results hMPV was identi- fied in 59(24.1%)of the 245 specimens tested,hMPV infection alone accounted for 13.1% of the infections in the 452 chil- dren under study,The prevalence of hMPV was higher than other respiratory viruses in winter.The mean age of hMPV-infec- ted children(n=59)was 27.7 months.There was no significant difference between age groups in terms of the prevalence of hMPV(P＞0.05).There were no statistically significant difference in demographics and clinical symptoms between hMPV in- fection and other common respiratory virus infection.Genotyping for the hMPV M gene from 23 Shanghai patients showed two distinct hMPV genotypes.Sequence analysis of these hMPV M genes showed 82.8%-100% homology to the registered se- quence in GenBank.There was no significant difference in clinical characteristics between the 2 genotypes.Conclusions hMPV plays an important pathogenic role in lower respiratory tract infection of children,hMPV prevailed in the winter of 2004.Clini- cally,hMPV infection can not be discriminated from the infection of other respiratory viruses.Clinical manifestation is similar between the two hMPV genotypes.

There are 250 species of Zanthoxylum (Rutaceae) in the world. This genus distributed in tropical and subtropical regions. Alkaloids are the major and representative ingredients in these plants including quinolines, isoquinolines, and amide alkaloids, with such biological activities as anti-tumor, anti-inflammatory, analgesic, anti-virus, anti-platelet aggregation, anti-bacteria and anti- oxidant. These species have been used for a long time to treat toothache, urinary and venereal diseases, lumbago and rheumatism. This review summarizes the chemical constituents and pharmacological activities from the Z. sppplants, in an effort to the systematic research and application of the alkaloids of this genus.
Alkaloids ; chemistry ; pharmacology ; Animals ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Molecular Structure ; Zanthoxylum ; chemistry

e: Objective:To establish a useable animal model for the purification and immunotherapy of HSP with the observation of the growth rule of S180 sarcome tumor tissue and the immunohistochemical expression of different HSP in mice. Methods:The tumor incidence?the role of growth?the survival time and immunohistochemical expression of different HSP were observed after the S180 sarcome cells were inoculated at Balb/C mice back subcutaneous in different dose. Results:Sarcoma were formed in 100%.There are significant difference between vary dose in survival time, The expression was positive in HSP60?70?90? and grp94, and HSP70 expression was enhanced whereas HSP90? expression was decreased after heat shock treated. Conclusion:The use of S180 sarcome in mice is a good model to observe the expression of different HSP and the tumor tissue is suitable to purify. The model can be also used in the library study of HSP tumor vaccine.

The kinetics of nicotine degradation by O.intermedium DN2 and its application in tobacco waste were investigated. The results showed that the optimum temperature of nicotine degradation by O.intermedium was 30 ℃, the pH value was 6.5 and a mount of inoculum was 5 %. Under above conditions, the kinetics of nicotine degradation of initial concentration 500 mg/L was studied. The results indicated that the degradation process of nicotine with no-induced strain DN2 followed inverse S-shaped curve, and degradation process of nicotine with induced cells of DN2 followed Eckenfelder mode. The half life of nicotine degradation was 17.43 h and 4.10 h, respectively. And the results also showed that tolerance of O.intermedium DN2 to nicotine was up to 5000 mg/L when 0.1 % of glucose was added. Nicotine (2 220 mg/L) in extract of tobacco wastes degraded about 95.22 % by strain DN2 in 60 h incubation, indicating that strain DN2 was of application value in treatment nicotine pollution.

BACKGROUND: Elimination of antigenic substances from natural extracellular matrix with the integrity of the tissue structure retained renders the matrix to possess better biocompatibility and provides a cell culture environment close to conditions of the internal environment. Such materials are the primary choice for cell culture scaffold in tissue engineering.OBJECTIVE: To prepare human articular cartilage acellular matrix so as to provide a methodological basis for further study of articular cartilage acellular matrix as cell scaffold materials.DESIGN: A single sample study of bone tissues.SETTING: The experiment was performed in Institute of Orthopedics, General Hospital of PLA, between January and May in 2004. The specimens were obtained from patients requiring joint replacement for femoral neck fracture.MATERIAIS: The experiment was conducted in the Department of Orthopedics, General Hospital of PLA from January to May in 2004. Human articular cartilage specimens were obtained from the femoral head of patients with total hip arthroplasty for femoral neck fracture.METHODS: Totally 10 specimens of fresh articular cartilage(3.5 mm × 4. 5 mm × 2.0 mm) were obtained and freeze-dried for 12 hours. Cartilage acellular matrix was prepared using Triton X-100, Dnase and Rnase and identified by means of hematoxylin-eosin(HE) and safranine O staining and immunohistochemical staining for cartilage proteoglycan.MAIN OUTCOME MEASURES: Histological observation of the articular cartilage acellular matrix and immunohistochemical staining of cartilage proteoglycan.RESULTS: HE and safranine O staining both showed no cellular structure in the matrix with only recesses left by the removed cells. Immunohistochemical staining for cartilage proteoglycan yielded positive results, suggesting the presence of cartilage proteoglycan in the acellular matrix.CONCLUSION: Human articular cartilage acellular matrix can be prepared using the modified four-step procedures with detergent and enzymatic extraction with lyophilization, and the preserved cartilage proteoglycan in the material may retain good pressure resistance.

Objective To evaluate the serum pyridinoline cross-linked carboxyteminal telopeptide of type Ⅰ collagen ( ICTP) in the early diagnosis potency,efficacy and prognosis of tumor bone metastasis. Methods According to emission computed tomography(ECT) ,MRI and X-ray results,336 cases of tumor were divided into higher ICTP (5. 98 ± 1. 95μg/L ) than normal values. Twenty-two cases were identified bone metastasis through PET/CT examination. 26 cases were identified bone the effective cases decreased from( 13. 22 ± 4.65)μg/L (before treatment) to (7. 18 ±3. 54)μg/L (after treatment) (t = 10. 076,P = 0. 000). Conclusions Serum ICTP is helpful for the early diagnosis, screening and efficacy evaluation of tumor bone metastasis. It could be used for monitoring the occurrence of tumor bone metastasis and its prognosis.