BACKGROUND: Recently, a few studies have reported the correlation between transforming growth factor-α (TGF-α) and graft-versus-host disease (GVHD); however, the combination of TGF-α with other cytokines in patients with chronic or acute GVHD requires further study.OBJECTIVE: To analyze the changes of serum tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4), and transforming growth factor-α (TGF-α) in leukemic patients after allogeneic hematopoietic stem cell transplantation (Allo-HSCT) and investigate the effects of these cytokines on different grades of GVHD.DESIGN: Case control study.SETTING: Department of Hematology, Organ Transplantation Center, the Second Affiliated Hospital, General Hospital of Chinese PLA; Department of Nuclear Medicine, the Second Affiliated Hospital, General Hospital of Chinese PLA.PARTICIPANTS: Forty-two leukemic patients (23 males and 19 females, 16-68 years old, mean age of 35 years) who underwent Allo-HSCT for the first time were selected from the Department of Hematology, Organ Transplantation Center, the Second Affiliated Hospital, General Hospital of Chinese PLA and Department of Transplantation, the 307 Hospital of Chinese PLA from June 2005 to June 2007. Twelve patients had acute granulocytic leukemia (AGL), fifteen patients had acute lymphocytic leukemia (ALL), and fifteen patients had chronic granulocytic leukemia (CGL). Among the 42 patients, 37 underwent peripheral blood transplantation and five received bone marrow transplantation. Twenty-one patients had acute GVHD (18 cases in grades Ⅰ-Ⅱ and three cases in grades Ⅲ-Ⅳ) after Allo-HSCT, but the other 21 patients did not. Fourteen patients had chronic GVHD (five cases of limited type and nine cases of extensive type), but the other 28 patients did not. An additional 30 healthy subjects (18 males and 12 females, 20-70 years old, mean age of 44 years) were collected as a normal control group. All patients provided confirmed consent, and the study was approved by the local ethics committee.METHODS: Levels of serum TNF-α, IL-4, and TGF-α in leukemic patients with Allo-HSCT and normal subjects were measured by radio-immuno-assay, the cytokines levels of the patients with/without acute GVHD, of those with/without chronic GVHD and of different grades of GVHD were compared.MAIN OUTCOME MEASURES: Comparisons of serum TNF-α, IL-4, and TGF-α among the groups.RESULTS: All 42 leukemic patients and 30 healthy subjects were included in the final analysis. Levels of TNF-α, IL-4, and TGF-α in patients with acute or chronic GVHD were significantly higher than those in the normal subjects (P<0.05-0.01). Levels of TNF-α and IL-4 in patients without acute GVHD were significantly higher than those in the normal subjects (P<0.01,0.05). Levels of TNF-α, IL-4, and TGF-α in patients with acute GVHD were significantly higher than those in patients without acute GVHD (P<0.05). Levels of TNF-α, IL-4, and TGF-α in patients with chronic GVHD were significantly higher than those in patients without chronic GVHD (P<0.05). Levels of serum TNF-α and TGF-α in patients with acute GVHD of grades Ⅲ-Ⅳ or chronic GVHD of extensive type were significantly higher than those in patients with acute GVHD of grades Ⅰ-Ⅱ or chronic GVHD of limited type (P<0.05-0.01).CONCLUSION: After Allo-HSCT, dynamically monitoring changes of levels of TNF-α, IL-4, and TGF-α may serve as a possible means of predicting the onset of acute or chronic GVHD and may contribute considerably to deciding clinical severity of GVHD.

BACKGROUND:Many scholars have experimental y confirmed the obvious effect of mesenchymal stem cells to repair radiation injury. OBJECTIVE:To preliminarily investigate the mechanism of human umbilical cord mesenchymal stem cells promoting the healing of combined radiation-wound skin injury and whether they possess tumorigenicity in vitro. METHODS:Fifteen Sprague-Dawley rats were randomly divided into three groups, five rats in each group. The right buttock of rats (2.5 cm×2.0 cm) was irradiated with 40 Gyβ-rays produced by a linear accelerator, in which a round wound with a diameter of 1.5 cm was made. After 12 hours of modeling, human umbilical cord mesenchymal stem cells at three concentrations (5.0×106, 1.0×107 and 2.0×107 ) were injected through tail vein of rats, and luciferin (20 mg/kg) was injected intraperitoneal y. celldistribution in vivo was traced using IVIS in vivo imaging system. The ability of human umbilical cord mesenchymal stem cells to form colonies was observed using the colony formation assay with soft agar. RESULTS AND CONCLUSION:Human umbilical cord mesenchymal stem cells injected through tail vein of rats were mostly gathered in the lungs. cells were accumulated in the injured site of rats injected with 2.0×10 7 human umbilical cord mesenchymal stem cells;however, the fluorescence signal was not observed in the injured site of rats injected with 5.0×106 and 1.0×107 human umbilical cord mesenchymal stem cells. The other results indicated human umbilical cord mesenchymal stem cells of low dose, medium dose and high dose had no colony formation on soft agar, but the tumor cells had a great ability to form colony. These findings indicate that human umbilical cord mesenchymal stem cells promote healing combined radiation-wound skin injury by local migration and exhibit no tumorigenicity in vitro in a short period.

BACKGROUND: Anterior cervical decompression and bone graft fusion is always the standard operation type for the treatment of cervical spondylotic radiculopathy and myelopathy. However, secondary degeneration and new symptoms caused by the stress increase of adjacent segments following anterior cervical spinal fusion have been paid more and more attention. Artificial cervical disc replacement provides a new choice for the treatment of cervical syndrome.OBJECTIVE: To observe the therapeutic effect of BRYAN& artificial cervical disc displacement for the treatment of cervical spondylotic radiculopathy and myelopathy.DESIGN: Self-control experiment.SETTING: Department of Orthopaedics, Sixth Hospital of Ningbo.PARTICIPANTS: Seventeen patients with cervical spondylotic radiculopathy and/or myelopathy admitted to the Department of Orthopaedics, Sixth Hospital of Ningbo were involved between December 2003 and February 2005 were involved in the study. Informed consents were obtained, and all participates were approved by Medical Ethics Committee.There were 9 patients with cervical spondylotic myelopathy, and 8 patients with cervical spondylotic radiculopathy.BRYAN& artificial cervical disc system is an artificial cervical intervertebral disc with composite structure. It is constructed by two titanium alloy end plates and polymer core, which can bear loading and took polyisocyanate as matrix.METHODS: BRYAN& artificial cervical disc system was used in the treatment. All the patients underwent single-segment displacement. Two patients underwent the displacement at C3-4, five at C4-5, 8 at C5-6 and 2 at C6-7. At postoperative 1, 3and 6 months, X-ray lateral position, anteflexion position, posterior extension position, the left and right side dynamic position plains were taken to investigate prosthetic stability. Neurologic function was evaluated according to Japanese Orthopaedic Association (JOA) scoring system.MAIN OUTCOME MEASURES: ① Investigating whether or not prosthesis displaced by means of dynamic position plains.② Investigating the recovery of neurologic function by JOA scoring system.RESULTS: ①JOA average scores were elevated to 15 from preoperative 8.5. Nervous system symptoms had improved to different extents, and the mean improvement rate was 75%. At postoperative 1,3 and 6 months, all the patients were followed up and their X-ray plains of cervical lateral position and flexion-extension position showed good stability,indicating that motor function was not lost obviously. No heterotopic ossification was found around the prothesis. ②Wound infection was not found in any patient. At postoperative 3 months, less than 2 mm slight displacement appeared in the prothesis in one patient, and there were no obvious symptoms. Prothesis abrasion, subsidence, bone bridge formation and other rejections were not found in the other prothesis. Prothesis had good biocompatibility, and could be examined by routine CT and MRI.CONCLUSION: Artificial cervical disc replacement can obviously improve the symptoms of patients, and has good therapeutic effects in the treatment of cervical syndrome.

Objective:To investigate the effect of transforming growth factor (TGF- β) signal in muscle fiber itself during inflammation/immunity response on intramuscular inflammation. Methods:Sixteen wild C57BL/6 mice (wild group) and sixteen mice with skeletal muscle-specific deficiency of T βRⅡ (knock-out group) between 4-8 weeks of age were selected for this study. Acute muscle injury in mice was induced by injection of myotoxin cardiotoxin (CTX) into gastrocnemius. The differences in intramuscular inflammation were compared between the wild and knock-out groups on 0, 4, 7 and 10 d after CTX injection by observing exudation of mononuclear phagocytes, macrophages, M1 type macrophages, CD4 +T cells and helpers T cells (Th1, 2&17). Two newborn C57BL/6 wild mice and 2 SM TGF- βr2-/- knock-out mice were selected to culture primary myoblasts in vitro which were divided into 2 groups: an interferon group subjected to interferon simulation and a control group subjected to addition of an equal amount of solvent. The differences in expression of IL-6, IL-10, MCP-1, MIP-1α, H-2K b, H2-Ea, Toll-like receptor (TLR)3 and TLR7 were compared between the interferon and control groups, as well as between the wild and knock-out groups. Results:On 4&7 d after CTX injection, the ratios of mononuclear/macrophage (75.73%±3.62%, 45.27%± 2.32%), macrophages (38.67%±2.76%, 24.87%±2.19%), M1 macrophages (43.21%±0.11%, 30.43%±2.19%), CD4 +T cells (20.13%±1.62%, 5.67%±0.32%) in the muscle tissue from the knock-out mice were significantly higher than those from the wild mice (58.52%±2.43%, 29.21%±2.45%; 20.63%±2.32%, 16.23%±1.25%; 24.98%±0.35%, 14.23%±1.69%; 10.70%±0.43%, 2.50%±0.45%), with a majority of Th1&Th17 ( P<0.05). In vitro results showed that the levels of IL-6, MCP-1, MIP-1α, H-2K b, H2-Ea and TLR3 were significantly upregulated in the interferon group compared with the control group and that such upregulation in the nock-out mice was more significant than in the wild mice ( P<0.05). Conclusions:Endogenous TGF- β signal activation plays a role in the functional recovery after muscle trauma, because it is involved in the regulation of immune behavior of muscle fibers, thus affecting intramuscular inflammation and muscle regeneration.

Objective To explore the value of combined detection with MMP-9 and uPA in the progno-sis of pancreatic carcinoma. Methods By immunohistochemistry PV methods,the expression of MMP-9 and uPA was respectively studied in 63 surgical specimens of primary pancreatic carcinoma and the survival time of patients with pancreatic carcinoma was analysed. Results The expressions of MMP-9 and uPA were positively related(r=0. 573,P=0. 000). The expression of MMP-9 and uPA significantly correlated with differentiation (r= -0. 271,P=0. 032;r= -0. 333,P=0. 008),TNM stages(r= -0. 449,P=0. 000;r= -0. 430,P=0. 000)and lymph node metastasis(r=0. 329,P=0. 009;r=0. 400,P=0. 001),separately. The expression of MMP-9 had also a significant correlation with tumer size(r= -0. 297,P=0. 018)and distant metastasis(r=0. 320,P=0. 011). Univariate analysis identified that tumor size(χ2 =8. 766,P=0. 012),differentiation(χ2 =29. 050,P=0. 000),clinical stage(χ2 =24. 940,P=0. 000),distant metastasis(χ2 =12. 846,P=0. 000), lymph node metastasis(χ2 =15. 457,P=0. 000),MMP-9(χ2 =32. 700,P=0. 000)and uPA(χ2 =41. 495,P=0. 000)were significantly associated with prognosis. Kaplan-Meier survival analysis showed that 1-year survival rate of patients with MMP-9 ( -),uPA ( -)were significantly longer than that of the patients with MMP-9( ﹢),uPA( ﹢),respectively(χ2 =32. 700,P=0. 000;χ2 =41. 495,P=0. 000);1-year survival rate of patients with MMP-9( -)/uPA( -)was significantly longer than the others( Log-rank test,χ2 = 54. 892, P=0. 000). COX regression revealed that differentiation(RR=2. 315,P=0. 004),clinical stage(RR=1. 694, P=0. 002),MMP-9(RR=0. 165,P=0. 000)and uPA(RR=0. 244,P=0. 007)was independent prognostic factors in pancreatic carcinoma. Conclusion They may have a synergistic function in the the process of growth and invasion in pancreatic cancer between MMP-9 and uPA,and the posssible mechanism is that uPA activate degradation of MMP-9,which is not favorable to prognosis. Combined analysis of MMP-9 and uPA may lead to a more reliable prognostic estimation,as the beneficial supplement of the differentiation,and clinical stage to judge the prognosis of pancreatic cancer.

MicroRNA has speciifc biological and expression properties in transcription level, which has more potential than genomic DNA in the identification for forensic body fluid, species and degraded crime biological materials. Here we introduce the speciifc research of forensic body lfuid identiifcation, research strategies and applied perspectives with forensic miRNA, expecting to provide the application and study of miRNA analysis for reference.

Objective To analyze the early risk factors of overweight and obesity in preschool children.Methods Using stratified cluster sampling,the data of 1 335 preschool children's physical examination in High-tech Zone,Urumqi,Xinjiang were collected,and the case group had 153 overweight and obese children,the control group had 1 182 non-overweight and obese children;a case-control study was conducted.The basic data of mothers and the basic data of neonatal birth were analyzed retrospectively.The univariate and unconditional multivariate logistic regression analysis was performed.Results The prevalence of overweight and obesity in preschool children in High-tech Zone in Urumqi was 11.5％.Non-conditional multivariate logistic regression analysis showed that children's age (OR=1.31,95％ CI:1.07-1.61),mother's pre-pregnancy BMI (OR=1.11 95 ％,CI:1.06-1.17) and whether mothers had gestational hypertension (OR=1.99 95％,CI:1.03-3.85) were the risk factors for overweight and obesity in preschool children (P＜0.05).Conclusion In Urumqi high school district preschool children's overweight and obesity rate was high;mothers with high BMI before pregnancy,and those with high blood pressure during pregnancy can increase the risk of overweight and obesity in children,preschool children's increased age may increase the risk of overweight and obesity in children.

OBJECTIVETo determine whether the synthesis of proteoglycan, collagen and associated ultrastructure are related to the adenovirus-mediated gene transferred to adult degenerative cells.

METHODSAdenovirus/cytomegalovirus human transforming growth factor-beta 1 (Ad/CMV-hTGF-beta 1) was used to transfect degenerative cells. Antonopulos method, Miamine method and transmission electron microscopy were conducted to study the synthesis of proteoglycan, collagen, and ultrastructure, respectively. Cell cultures were established from the nucleus pulpous and annulus fibrosus tissues, which were taken from surgery.

RESULTSNucleus pulpous and annulus fibrosus cells were efficiently transduced by the adenoviral vector carrying hTGF-beta 1 gene. The synthesis of proteoglycan and collagen increased compared with the control group (P < 0.05). The metabolism of cells was slightly improved. No significant toxic effects were found.

CONCLUSIONSExpression of hTGF-beta 1 gene is efficient to accelerates proteoglycan synthesis and thus accelerates the improvement of collagen. The function and structure of degenerative cells are improved. Ad/CMV-hTGF-beta 1 may be suitable for treating disc degeneration.

Adenoviridae ; Adult ; Cells, Cultured ; Collagen ; biosynthesis ; Extracellular Matrix ; metabolism ; Genetic Vectors ; Humans ; Intervertebral Disc ; ultrastructure ; Proteoglycans ; biosynthesis ; Spinal Diseases ; metabolism ; pathology ; Transfection ; Transforming Growth Factor beta ; genetics ; Transforming Growth Factor beta1

OBJECTIVETo provide a highly efficient adenoviral vector Ad-CMV-hTGFbeta1 for the study of gene therapy for reversion of the intervertebral disc degeneration.

METHODSA newly developed recombinant adenoviral vector construction system was used in the study. The cDNA of hTGFbeta1 was first subcloned into a shuttle plasmid pShuttle-CMV. The resultant plasmid was linearized by digesting with restriction endonuclease PmeI, and subsequently transformed into E.coli. BJ5183 cells with an adenoviral backbone plasmid pAdEasy-1. Recombinants were selected by kanamycin resistance and confirmed by restriction endonuclease analysis. Finally, the recombinant plasmid linearized by PmeI was transfected into 293 cells. Recombinant adenoviruses were generated within 2 weeks.

RESULTSThe recombinant adenoviral plasmids were cut by BamHI and PacI respectively, and the diagnostic fragments appeared in 0.8% agarose electrophoresis. The infected 293 cells showed evident cytopathic effect (CPE). The productions of PCR confirmed the presence of recombinant adenovirus. The expression of hTGFbeta1 was verified by immunohistochemical staining.

CONCLUSIONSThe successful generation of the adenoviral vector Ad-CMV-hTGFbeta1 and the confirmation of the interest gene expression make it possible for the experimental study of the reversion of the intervertebral disc degeneration by gene therapy.

Adenoviridae ; genetics ; Base Sequence ; Cells, Cultured ; Cytomegalovirus ; genetics ; Gene Expression Regulation ; Gene Transfer Techniques ; Genetic Vectors ; genetics ; Humans ; Immunohistochemistry ; Intervertebral Disc ; cytology ; pathology ; Molecular Sequence Data ; Polymerase Chain Reaction ; Recombination, Genetic ; Sensitivity and Specificity ; Transforming Growth Factor beta ; genetics ; Transforming Growth Factor beta1

BACKGROUNDSteady-state bone marrow (SS-BM) and granulocyte colony-stimulating growth factor-primed BM/peripheral blood stem-cell (G-BM/G-PBSC) are the main stem-cell sources used in allogeneic hematopoietic stem-cell transplantation. Here, we evaluated the treatment effects of SS-BM and G-BM/G-PBSC in human leucocyte antigen (HLA)-identical sibling transplantation.

METHODSA total of 226 patients (acute myelogenous leukemia-complete remission 1, chronic myelogenous leukemia-chronic phase 1) received SS-BM, G-BM, or G-PBSC from an HLA-identical sibling. Clinical outcomes (graft-versus-host disease [GVHD], overall survival, transplant-related mortality [TRM], and leukemia-free survival [LFS]) were analyzed.

RESULTSWhen compared to SS-BM, G-BM gave faster recovery time to neutrophil or platelet (P < 0.05). Incidence of grade III-IV acute GVHD and extensive chronic GVHD (cGVHD) was lower than seen with SS-BM (P < 0.05) and similar to G-PBSC. Although the incidence of cGVHD in the G-BM group was similar to SS-BM, both were lower than G-PBSC (P < 0.05). G-BM and G-PBSC exhibited similar survival, LFS, and TRM, but were significantly different from SS-BM (P < 0.05). There were no significant differences in leukemia relapse rates among the groups (P > 0.05).

CONCLUSIONSG-CSF-primed bone marrow shared the advantages of G-PBSC and SS-BM. We conclude that G-BM is an excellent stem-cell source that may be preferable to G-PBSC or SS-BM in patients receiving HLA-identical sibling hematopoietic stem-cell transplantation.

Adolescent ; Adult ; Aged ; Bone Marrow ; drug effects ; Bone Marrow Transplantation ; methods ; Child ; Female ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; therapy ; Leukemia, Myeloid, Acute ; therapy ; Male ; Middle Aged ; Retrospective Studies ; Stem Cells ; cytology ; Young Adult