The article reviews concisely around the history of hematopoietic stem cell research, basic and clinical, since its very beginning after the nuclear explosions at the end of the Second World War, that explains why the stem cell study in the world began with the hematopoietic stem cells and the existence of non-hematopoietic stem cell in vivo had been left out of account for many decades till 21st century. During 50-60s of the last century, it was known from the animal experiments that there must be hematopoietic stem cells existing in vivo and believed that the effective bone marrow transplant is actually the stem cell transplantation. It is reviewed how the basic studies of stem cell interacted with the clinical stem cell transplantation and how great the contribution was given to strongly push forward the development of contemporary stem cell biology and modern hematology from the basic studies especially in the immunological and molecular biological fields, for instance, the applications of HLA technology and monoclonal antibody produced, flow cytometry, and genetic recombinant cytokines, the novel technique for gene cloning, genomics, proteomics, and iRNA as well as bioinformatics. It has lead to the pluralistic cell therapy as a novel trend in stem cell transplantation as to combine immunotherapy and mesenchymal stem cells with the conventional stem cell transplants. This paper looks back in the past several decades, however, on every achievement of stem cell study that were usually accompanied with some idealistic one-sidedness or even errors in design and conclusion of some experiments. Usually it took a period of 2 or even 4 decades to clarify some basic idea, that seemed normal in the science development, for examples, the dividing line between the hematopoietic stem cell and progenitor cells, possibility to expand or clone the real stem cell ex vivo, and whether the majority of leukemias are originated from stem cell level, etc. Towards the end of 20th century, the greatest discovery was the existence of adult stem cells in adult tissues, which are no doubt the remnants from the embryonic development including all stages of embryonic stem cells, very earlier and later, hematopoietic and nonhematopoietic, and could be induced to differentiate into all kinds of tissue cells by proper ways in vitro. No evidence has really provided for the hypothesis of so called trans-differentiation or de-differentiation, which brought about strong calls in questions in the past 5 years. The problems in developing the clinical gene therapy by using hematopoietic stem cell as carrier of interested gene still remained to be solved so far. Because of the relatively weak base of related basic studies, the clinical application of gene therapy resulted in the failure of clinical practice with lots of lessons towards the end of last century. The whole history of stem cell study in the world was an endless process of continuous redress for theoretical ideas in stem cell biology that had never been consummate.
Hematopoietic Stem Cells ; cytology ; History, 19th Century ; History, 20th Century ; History, 21st Century ; Research ; history ; standards ; Research Design ; standards

Abortion, Missed ; chemically induced ; Adult ; Carbon Disulfide ; adverse effects ; Female ; Humans ; Occupational Exposure ; adverse effects ; Pregnancy ; Retrospective Studies ; Risk Factors ; Time Factors

Objective To compare four methods of monitoring vascularization of tissue engineered bone in the rhesus so as to find our the best. Methods Twenty-five lower limbs of 13 rhesuses were used in this study to make models of tibial diaphyseal defect of 20mm which were to be fixed with an AO reconstruction plate of 7 holes. The monkeys were randomly divided into five groups according to defect filling materials: group A:?-tricalcium phosphate (?-TCP) and bone marrow stromal cells (BMSCs) and blood vessel bundles; group B:?-TCP and blood vessel bundles; group C:?-TCP and BMSCs; group D:?-TCP; group E: blank. Perfusion weighted MR imaging (PWMR), X-ray, radionuclide imaging and histological examinations were carried out at weeks 4, 8, 12 postop- eratively. The maximum slope rates of the single intensity-time curve (SS_(max)) and values of baseline (Sl_(?))were calculated at the same time points. Transmittances of the X-ray films were assessed. Ratios between isotope counts in region of interest (ROI) were calculated. Chinese ink perfusion and calculation of blood vessel areas were done for histological examinations, Results Compared with other groups, the SS_(max) in group A was the highest at weeks 4, 8, 12 postoperatively. In group A, the SS_(max) at week eight was significantly higher than that at week four (P= 0. 003), and the SS_(max) and transmittance of X-ray were negatively related at week 12 after operation (rs=-0. 892, P=0. 042), but the SS_(max) and blood vessel area were positively related (rs=0. 894, P=0.041)Conclusions PWMR can be a sensitive, quantitative, noninvasive and non-radiant method to monitor vascularization of tissue engineered bone, because SS_(max) of the single intensity-time curve of PWMR can reflect the most accurately the process of vascularization of tissue engineered bone.

Objective To explore the effect of cutaneous never anastomosis on sensory recovery in repairing wide spreadly soft tissue defects in dorsal hand with free anterolateral femoral flap. Methods The cases with wide spreadly soft tissue defects in dorsal hand repaired with free anterolateral femoral flap from January 2006 to December 2012 were divided into 2 groups. The control group including 15 consecutive patients from January 2006 to January 2009, whose sensation was reconstructed in routine way. Other 15 consecutive patients from Febnary 2009 to December 2012 were as research group, whose sensation was reconstructed with the suture of cutaneous nevers of anterolateral femoral flaps and forearm. All the patients were followed up for 12~24 months. Results All the free flaps survived in both groups. There were 4 cases good of sensory recovery in the control group, and it was 11 in the research group. No ulceration happened. Conclusion Cutaneous never anastomosis may result in satisfactory sensory function in the patients with wide spreadly soft tissue descts in dorsal hand repaired with free anterolateral femoral flap.

Objective To investigate the clinical value of [11C]CFT PET in the diagnosis and severity assessment of Parkinson disease (PD). Methods Thirty-eight patients with PD at various Hoehn & Yahr (H&Y) stages were included and underwent a [11C]CFT PET scan. The correlation between [11C]CFT uptake and unified Parkinson disease rating scale part III (UPDRS III) of PD patients was evaluated by calculating Pearson’s regression coefficient. Statistical parametric mapping (SPM) analysis was performed to compare the difference of dopamine transporter (DAT) distribution between ear-ly and advanced PD patients. Results There was a significant reduction of [11C]CFT uptake in the bilateral striatum of PD patients. There was a significant negative correlation between clinical scores of UPDRS III, rigidity, bradykinesia, pos-ture, gait and [11C]CFT uptake in the striatum. The SPM analysis revealed a significant and asymmetric decrease of [11C] CFT uptake in the striatum, predominantly on the putamen and caudate nucleus contralateral to the onset limb, in the posterior area of ipsilateral putamen in early PD (H&Y 1-2) patients compared with the normal controls. There was a sig-nificant symmetric decrease of [11C]CFT uptake in both putamen and caudate nucleus in advanced PD (H&Y 3-5) pa- tients, compared with normal controls. Compared with early PD patients, the reduction of DAT was more severe in bilater-al caudate nucleus and the ipsilateral putamen in the advanced PD patients. Conclusions [11C]CFT PET is a sensitive biomarker in the diagnosis and assessment of disease severity of PD patients.

OBJECTIVETo observe the long-term effect of tissue engineering-based repair of large weight-bearing bone defect in goats, and the final outcome of the scaffold material coral hydroxyapatite (CHAP) in vivo.

METHODSFifteen Chinese goats were subjected to operations to induce a 2-cm left tibial diaphyseal defect, which was filled subsequently with CHAP and bone marrow stromal stem cells (BMSCs). The repaired defects were evaluated by ECT, X-ray and histology in the early stage and at 6, 12, 18, and 24 months postoperatively.

RESULTSECT showed good bone regeneration and revascularization within 2 months postoperatively. X-ray and histology displayed eccentric and gradual bone regeneration in the early stage, and the tissue-engineered bone graft was firmly healed with the goat tibia. X-ray and histological examination at 6, 12, 18, 24 months postoperatively revealed moulding of the new bones and medullary cavity recanalization, and the structure of CHAP disappeared and gradually integrated into the new bones.

CONCLUSIONTissue-engineered bone is capable of total repair of large bone defect in goats by forming normal functional new bones. CHAP can be eventually degraded completely and become the component of the newly generated bones.

Animals ; Bone Marrow Transplantation ; Bone Regeneration ; Bone Substitutes ; Cells, Cultured ; Goats ; Prostheses and Implants ; Tibial Fractures ; physiopathology ; surgery ; Tissue Engineering ; methods

Total body irradiation combined with cyclophosphamide (TBI/CY) and busulphan combined with cyclophosphamide (BU/CY) are standard conditioning regimens in hematological stem cell transplantation for patients with myelogenous leukemia. This study was aimed to compare the therapeutic efficacy of TBI/CY and BU/CY as conditioning regiment for acute or chronic myelogenous leukemia. The Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, CNKI, CBM (Chinese Bio-medicine Database) had been searched for all relevant articles (1999-2007). Comparative studies were carried out on clinical therapeutic effects of TBI/CY and BU/CY including stem cell engraftment, relapse, complications, transplant-related mortality, and disease-free survival. A meta-analysis was performed using Review Manager 4.2 software and funnel plot regression was adopted to assess the publication bias. The results indicated that 2149 articles in English and 46 articles in Chinese were got, and finally 9 clinical trials with total 3039 patients have been assessed. No significantly difference was found in engraftment failure and transplant-related mortality resulting from TBI/CY and BU/CY conditioning regimens, but the incidence of veno-occlusion of liver and hemorrhagic cystitis obviously increased in BU/CY group after transplantation, the acute GVHD, interstitial pneumonia and cataract significantly increased in TBI/CY group. The relapse rate of AML in TBI/CY group was lower than that in BU/CY group, and the rate of long-term disease-free survival of AML patients in TBI/CY group also significantly lower than that in BU/CY group, but the relapse rate of CML in TBI/CY group after transplantation was obviously higher than that in BU/CY group, but there was no difference in longterm disease-free survival rate between the two conditioning regimens mentioned above. It is concluded that the meta-analysis confirms different effects of TBI/CY and BU/CY regimens on myelogenous leukemia transplantation. This result is useful for physicians to select treatment regimens.
Busulfan ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Disease-Free Survival ; Humans ; Leukemia, Myeloid ; radiotherapy ; surgery ; Leukemia, Myeloid, Acute ; radiotherapy ; surgery ; Transplantation Conditioning ; methods ; Treatment Outcome ; Whole-Body Irradiation

OBJECTIVETo study the clinical outcome, adverse effect and treatment cost of homoharringtonine (HHT) in combination with all-trans retinoic acid (ATRA) and arsenic trioxide (AS2O3) for newly diagnosed with patients acute promyelocytic leukemia (APL).

METHODSClinical data of treatment of newly diagnosed patients with APL in experimental group (HHT + ATRA + AS2O3, n = 14) and control group \[Idarubicin (IDA) + ATRA + AS2O3, n = 21\] were analyzed retrospectively. The therapeutic effects, side effects and costs during induction therapy were compared between the two groups.

RESULTS(1) The complete remission (CR) rate were 92.9% (13/14) and 95.2% (20/21) in experimental group and control group, respectively. The time to achieve CR were (28.1 ± 3.8) and (31.7 ± 4.2) days, respectively (P > 0.05). The negative rate of PML-RARα fusion gene at the time of CR were 76.9% (10/13) and 75.0% (15/20), respectively, and that in CR patient at the end of the first cycle treatment were 100.0% (13/13) and 95.0% (19/20), respectively (P > 0.05). (2) 5-year overall survival (OS) rate were (92.6 ± 0.6)% and (89.9 ± 0.5)%, respectively (P > 0.05), 5-year disease free survival (DFS) rate were 100.0% and (86.8 ± 0.6)%, respectively (P > 0.05). (3) During induction therapy, the incidence of infection in experimental and control group were 23.1% (3/13), 60.0% (12/20), respectively (P < 0.05). The amount of platelet transfusion were (54.7 ± 29.6) and (76.5 ± 25.6) units, respectively (P > 0.05), and that of fresh frozen plasma were (1157.1 ± 238.4) and (1423.5 ± 324.6) ml, respectively (P > 0.05). The total medical costs (excluding HHT and IDA) in experimental and control group were (36074.9 ± 1245.6) and (50564.5 ± 3658.4)CNY, respectively (P < 0.05).

CONCLUSIONHHT in combination with ATRA and AS2O3 regimen for newly diagnosed APL has a better efficacy, a higher long-term survival rate, and a lower costs, which is one of the reasonable choice.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Arsenicals ; therapeutic use ; Female ; Harringtonines ; therapeutic use ; Humans ; Leukemia, Promyelocytic, Acute ; drug therapy ; Male ; Middle Aged ; Oxides ; therapeutic use ; Retrospective Studies ; Treatment Outcome ; Tretinoin ; therapeutic use

OBJECTIVETo study a new implant material (carbonated hydroxyapatite, CHA) united pedicle screw to cure spine fracture.

METHODSThirty-two cases of spine compressed fracture were used with pedicle screw fixator and vertebroplasty. Before operation, patients' vertebral body were compressed (46 + 21)% (20% approximately 70%) on average. In operation, broken vertebral body was reposition through pedicle screw technique, then used self-made syringe to inject CHA into anterior and central column of broken vertebral body through pedicle. And all of patients were not given any bone-graft.

RESULTSIn 6 - 26 months followed-up, no immunologic rejection was found about hydroxyapatite, and no any broken of the screws and shafts was found, no loosing and other complications either. All the patients could move in 3 - 5 days after operation. The height of the broken vertebral body were reduced 97% compared with pre-operation. And CHA in vertebral body was degraded gradually, and at the same time it was replace by new bone in vertebral body. After operation, VAS score was 61 +/- 32, and there was significant difference compared with pre-operation.

CONCLUSIONSThe pedicle screw fixation united vertebroplasty is an efficient way to prevent the failure of the treatment of spine fracture.

Adult ; Bone Screws ; Bone Substitutes ; therapeutic use ; Durapatite ; therapeutic use ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; instrumentation ; methods ; Fractures, Compression ; surgery ; Humans ; Male ; Middle Aged ; Spinal Fractures ; surgery ; Vertebroplasty ; methods

The production of recombinant protein is one of the major successes of biotechnology, animal cells are required to synthesize proteins with the appropriate post-translational modifications. Transgenic animal mammary gland bioreactor are being used for this purpose. Gene targeting is a more powerful method to produce mammary gland bioreactor, and nuclear transfer from cultured somatic cells provides an wonderful means of cell-mediated transgensis. Here we describe efficient and reproducible gene targeting in goat fetal fibroblasts to place the human tissue plasminogen activator mutant (ht-PAm) cDNA at the beta-casein locus, and would produce the transgenic goat by nuclear transfer. To construct the gene targeting vector pGBC4tPA, the milk goat beta-casein genomic DNA sequence for homologous arms had been cloned firstly. The left arm was 6.3 kb fragment including goat beta-casein gene 5' flanking sequence, and the right arm was 2.4 kb fragement including beta-casein gene from exon 8 to exon 9. The ht-PAm cDNA was subcloned in the goat beta-casein gene exon 2, and the endogenous start condon was replaced by that of ht-PAm. The bacterial neomycin (neo) gene as positive selection marker gene, was placed in the beta-casein gene intron 7, the thymidine kinase (tk) as the negative selection marker gene, was just outside the right arm. The validity of the positive-negative selection vector (PNS), was tested, and targeting homologous recombination (HR) were elevated to 5-fold with the negative selection marker using the drug GANC. The DNA fragment in which two LoxP sequence was delected effectively using Cre recombinase in vitro. Goat fetal fibroblasts were thawed and cultured to subconfluence before transfection, about 10(7) fibroblasts were electoporated at 240V, 600 microF in 0.8 mL PBS buffer containing linear pGBC4tPA. transfected cells were cultured in collagen-coated 96-wellplate for 24h without selection, then added the drug G418 (600 microg/mL) and GANC (2 micromol/L). After 12 days of selection, well separated G418r/GANCr clones were isolated and expanded in 24-wellplate. 244 clones were selected, and only 90 clones could grow and be tested by PCR screening for targeting. The primary result demonstrated that 31 targeting cell clones with homologous recombination events were obtained, and 2 cell clones was verified by DNA sequence analysis on the homologous recombination region.
Animals ; Animals, Genetically Modified ; genetics ; Base Sequence ; Caseins ; genetics ; Cloning, Organism ; DNA, Complementary ; genetics ; Gene Knock-In Techniques ; Genetic Engineering ; methods ; Genetic Vectors ; chemical synthesis ; Goats ; genetics ; Humans ; Mammary Glands, Animal ; cytology ; metabolism ; Molecular Sequence Data ; Mutant Proteins ; genetics ; Tissue Plasminogen Activator ; genetics