ObjectiveTo investigate the distribution of Kashin-Beck disease(KBD) in Tibet, and assess the disease status. Methods Between 2007 and 2008, a survey was done on KBDepidemiology which was carried out in four prefectures of 26 counties according to the east, south, west, north and center in Nakchu,Lhoca, Nyingtri and Shigatse districts of Tibet, with towns and villages as baseline survey points. According to the KBD e survey scheme, KBD clinical examination for adults was also carried out and at the same time clinical and right hand anteroposterior X-ray examinations were given to children aged 4 - 13. The partition of endemic area was based on the criteria of national standards for Kashin-Beck disease diagnoses《GB 16395-1996》. Slight KBD area:clinical prevalence of Kashin-Beck disease grade Ⅰ and above was less than 10％ or X-ray detection rate ＜ 10％ of children; the moderate prevalent KBD area: clinical prevalence of Kashin-Beck disease grade Ⅰ and above was between 10％ and 20％ or X-ray detection rate was between 10％ and 30％ of children; severe KBD area: clinical prevalence of KBD grade Ⅰ and above was more than 20％ or X-ray detection rate was higher than 30％ of children.ResultsA total of 108 townships of 26 counties were surveyed, 14 686 adults were clinically examined, cases detection of grade Ⅰ and above were 637 people, the prevalence was 4.34％, and no case of grade Ⅲ was detected.Of 5769 children's right anteroposterior X-ray film, 102 were detected positive; the prevalence rate was 1.77％.Metaphysis was affected in most of the child cases, which accounting for 89.2％ (91/102). Amongst all the counties, there were 10 counties, clinical detection rate of adult KBD was 0, and children's X-ray detection rate of KBD was also 0. In 1 county the clinical prevalence rate for adults KBD was 0 and X-ray detection rate for children was 3.66(7/191 ). In 12 counties the clinical prevalence rate for adults KBD was between 1.03％ and 7.54％, X-ray detection rate for children was between 0 and 7.76％, amongst all these counties surveyed there were 5 counties,the detection rate for children was 0. In 3 counties the clinical prevalence rate for adult KBD was between 10.69％and 13.88％, the X-ray detection rate for children was between 5.31％ and 7.76％. Conclusions According to the criteria for diagnoses of KBD, within the 26 counties surveyed, 10 counties are non-endemic areas, 13 counties are slight endemic areas, 3 counties are medium endemic areas. So far, KBD is prevalent in 52 counties of 7 prefectures (cities) in Tibet, the disease is widely distributed, the situation is still severe, and there is a need to continue to strengthen KBD surveillance.

OBJECTIVETo investigate the incidence and spermatogenesis of cryptorchid testes induced by postnatal injection of estradiol.

METHODSNinety male newborn Balb/C mice were randomly divided into an experimental (n = 60), a solvent control (n = 20) and a normal control group (n = 10). The experimental mice were again assigned to a 4-week, a 6-week, an 8-week, and a 10-week subgroup, and injected subcutaneously with 17-beta estradiol (5 microg/d) from 3 to 28, 3 to 42, 3 to 56 and 3 to 70 days after birth, respectively. The incidence of cryptorchidism and morphological changes of the testes were observed at 2 weeks after drug withdrawal.

RESULTSThe incidence rates of cryptorchidism in the 4-, 6-, 8- and 10-week groups were 0%, 26.7%, 60% and 60%, respectively, but no cryptorchidism occurred in the solvent and normal control groups. The 4- and 6-week groups showed autonomous descent of the cryptorchid testes and recovery of spermatogenesis after drug withdrawal. The models became stable and no spermatogenesis recovery was observed after 8 weeks of continuous medication.

CONCLUSIONStable cryptorchid infertile models can be established in mice by postnatally continuous injection of estradiol for over 8 weeks.

Animals ; Animals, Newborn ; Cryptorchidism ; chemically induced ; Disease Models, Animal ; Estradiol ; adverse effects ; Infertility, Male ; chemically induced ; Male ; Mice ; Mice, Inbred BALB C

Objective To explore the feasibility of rapid and sutureless anastomosis of artificial vascular replacement of abdominal aorta in dog models using magnetic compression anastomosis (MCA) technique. Methods Twelve healthy adult crossbred dogs were evenly divided into the MCA and hand suturing (HS) groups according to the anastomosis method between abdominal aorta and artificial blood vessels. The intraoperative duration of abdominal aorta occlusion, intraoperative condition of anastomotic stoma and postoperative imaging examination of anastomotic stoma were compared between two groups. Results The intraoperative duration of abdominal aorta occlusion in the MCA group was significantly shorter than that in the HS group [(5.2±2.3) min vs. (24.4±4.3) min, P < 0.001]. No anastomotic leakage of blood or anastomotic stenosis occurred in the MCA group during the operation. Intraoperative anastomotic leakage of blood occurred in all of the 6 dogs in the HS group. Among them, 1 dog died of excessive blood loss, and 2 dogs experienced mild anastomotic stenosis due to repeated repair. Postoperative color Doppler ultrasound and angiography showed smooth blood flow at the anastomotic stoma without stenosis or thrombosis in the MCA group. In the HS group, 4 dogs presented with anastomotic stenosis on angiography at postoperative 4 weeks. Conclusions MCA technique may achieve rapid and sutureless anastomosis of artificial vascular replacement of abdominal aorta in dog models, which reduces the incidence of anastomotic complications and accelerates postoperative recovery.

Traditional Tibetan medicine (TTM) is major component of traditional Chinese medicine (TCM). Over thousands of years, the Tibetan people have gradually established a comprehensive theoretical system for Tibetan medicine and obtained rich clinical experience in the snow-covered plateau. The history of TTM has developed rapidly, especially since the establishment of the People's Republic of China, and remarkable achievements have been made with the great support for ethnic medicines from the Communist Party of China and the central government. In this paper, we give an overview of the history, theoretical system, common treatment methods, current development, and prospects for the future development of TTM.

OBJECTIVETo investigate the effects of Tpo and/or IL-11 gene modified stromal cells on the expansion of CD(34)(+) hematopoietic stem/progenitor cells in cord blood.

METHODSRetroviral vectors containing Tpo or IL-11 gene were constructed and used to transfect the stromal cell line HFCL. Tpo and/or IL-11 mRNA was assayed by Northern blot. Non-modified stromal cells were used, CD(34)(+) hematopoietic stem/progenitor cells from cord blood were expanded on gene-modified stromal cells for 7 days. The phenotype of CD(34)(+)CD(38)(-) primitive progenitors was detected by flow cytometry.

RESULTSHFCL expressed Tpo and/or IL-11 mRNA after transfected by the retroviral vectors. The percentages of CD(34)(+)CD(38)(-) primitive progenitors in the cultures of Tpo, IL-11 and Tpo + IL-11 modified HFCL were (1.8 +/- 0.24)%, (1.62 +/- 0.23)%, and (2.45 +/- 0.28)%, respectively, which were higher than that in the control [(0.8 +/- 0.23)%].

CONCLUSIONThe stromal cells modified by Tpo and/or IL-11 gene were able to enhance ex vivo expansion of CD(34)(+) and CD(34)(+)CD(38)(-) hematopoietic stem/progenitor cells from cord blood.

ADP-ribosyl Cyclase ; analysis ; ADP-ribosyl Cyclase 1 ; Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; physiology ; Humans ; Infant, Newborn ; Interleukin-11 ; genetics ; Membrane Glycoproteins ; Stromal Cells ; physiology ; Thrombopoietin ; genetics

OBJECTIVETo identify genes that differentially expressed in Lin(-)CD(34)(-) and Lin(-)CD(34)(+) cells.

METHODSWith Lin(-)CD(34)(-) cells as tester and Lin(-)CD(34)(+) cells as driver, cDNA subtractive library for Lin(-)CD(34)(-) cells was constructed using suppression subtractive hybridization technique. Part of clones in the library were sequenced and the homologue analysis was conducted against the DNA database in GenBank.

RESULTS593 clones containing an average of 300 - 500 bp insert were identified. Of them, 53 randomly selected ESTs were sequenced. Homologue analysis revealed that 37 ESTs represented 10 known genes, and the other 16 ESTs represented 4 novel sequences.

CONCLUSIONPart of specifically expressed genes in Lin(-)CD(34)(-) cells were identified, which maybe related to Lin(-)CD(34)(-) cells' specific characteristics.

Antigens, CD34 ; metabolism ; Cloning, Molecular ; Gene Expression Profiling ; Gene Library ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; In Vitro Techniques ; Nucleic Acid Hybridization

UNLABELLEDTo identify differentially expressed genes between fetal mesenchymal stem cell (MSC) and adult MSC, especially specified genes expressed in fetal MSC, a cDNA subtractive library of fetal MSC was constructed using suppression subtractive hybridization (SSH) technique. At first, total RNA was isolated from fetal and adult MSC. Using SMART PCR synthesis method, single-strand and double-strand cDNAs were synthesized. After Rsa I digestion, fetal MSC cDNAs were divided into two groups and ligated to adaptor 1 and adaptor 2 respectively. Results showed that the amplified library contains 890 clones. Analysis of 890 clones with PCR demonstrated that 768 clones were positive. The positive rate is 86.3%. The size of inserted fragments in these positive clones was between 0.2 - 1 kb, with an average of 400 - 600 bp.

CONCLUSIONSSH is a convenient and effective method for screening differentially expressed genes. The constructed cDNA subtractive library of fetal MSC cDNA lays solid foundation for screening and cloning new and specific function related genes of fetal MSC.

Cloning, Molecular ; DNA, Complementary ; genetics ; metabolism ; Deoxyribonucleases, Type II Site-Specific ; metabolism ; Fetus ; Gene Library ; Humans ; Mesoderm ; cytology ; metabolism ; Polymerase Chain Reaction ; Stem Cells ; cytology ; metabolism

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Male ; Osteoarthritis ; epidemiology ; Tibet ; epidemiology

To investigate the expression of telomerase in cord blood hematopoietic stem/progenitor cells during their committed differentiation in vitro and provide an index of monitoring the proliferating potential of the hematopoietic stem/progenitor cells and security for clinical application. Human CD34 positive cells were isolated from umbilical cord blood by using magnetic cell sorting system (MACS), and were induced to differentiation with hematopoietic growth factors (SCF + IL3 + IL6 + GCSF and SCF + IL3 + IL6 + EPO) in a liquid culture system. The telomerase activity and the cytalytic subunit of telomerase (hTERT) of the cells were analysed during different periods of culture by using TRAP-PCR, TRAP-ELISA, Western blot and RT-PCR techniques, respectively. The results showed that a peak of cell growth was achieved on day 14 - 21 during induction of differentiation in vitro. Total cell number could increase 1006.4 +/- 103.2 times and could not increase there after. Telomerase activity and hTERT expression were low in freshly isolated cord blood CD34(+) cells and increased after about 7 days of culture in addition of cytokine combinations of SCF + IL3 + IL6 + GCSF and SCF + IL3 + IL6 + EPO, respectively. The telomerase activity and hTERT decreased after 14 days of culture and were not detected after 28 days of culture. It was concluded that the hematopoietic stem/progenitor cells can be expanded in large number in vitro and do not have the character of immortality and the telomerase activity could be a useful index in hematopoiesis regulation.
Antigens, CD34 ; blood ; Blotting, Western ; Cell Differentiation ; DNA-Binding Proteins ; Enzyme-Linked Immunosorbent Assay ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; enzymology ; Humans ; Polymerase Chain Reaction ; RNA, Messenger ; analysis ; Telomerase ; genetics ; metabolism

To evaluated the feasibility of preventing infection after high dose chemotherapy and radiotherapy using the granulocytes derived from differentiated from hematopoietic stem/progenitor cells ex vivo, human CD34-positive cells were isolated from umbilical cord blood by using a high-gradient magnetic cell sorting system (MACS), and the cells committedly differentiated with hematopoietic cytokines (SCF + IL-3 + IL-6 + G-CSF) in a liquid culture system. The expanded cell number, ratio of the viable cells, chromosome and phenotype of the differentiated cells and safety analysis of expanded cells were detected by using cell count, trypan blue exclusion test, karyotype analysis, flow cytometry and tumorigenic model of nude mice, respectively. The results showed that the combination of cytokines increased cell number by (1006.4 +/- 103.2) folds and flow cytometric analysis showed myeloid marker CD11b expressed in the about 60% cells. The growth peak of differentiated cells was at 14 days of culture and decreased at about 33 days. No abnormality was found in the karyotype analysis of expanded cells. No tumor was found in the nude mice injected with expanded cells after 35 days and the expanded cells had the ability of phagocytizing bacteria. It is concluded that the cells, differentiated from CD34(+) cells, expanded ex vivo possess the function of granulocyte and it was safe for clinical trial.
Animals ; Antigens, CD34 ; analysis ; Cell Differentiation ; Fetal Blood ; cytology ; Granulocytes ; cytology ; immunology ; Hematopoietic Stem Cells ; cytology ; Humans ; Karyotyping ; Mice ; Mice, Inbred BALB C ; Phagocytosis