The influences of E23K polymorphism of inwardly rectifying K~+channel 6.2 (Kir6.2) gene on the clinical phenotype of type 2 diabetes and glucose-lowering effect of gliclazide were investigated.The result showed that E23K polymorphism did not influence glucose-lowering effect of gliclazide,but serum creatinine level of patients with K/K genotype was higher than those of E/E and E/K genotypes (P

OBJECTIVETo investigate the best injection method of the bone marrow mesenchymal stem cells (BM-MSCs) transplantation for the treatment of a rat model with hind limb ischemia.

METHODSTwenty four SD rats with hind limb ischemia were randomly divided into four groups: control group, model group, acupoint BM-MSCs injection group (API group) and thigh muscle BM-MSCs injection group (TMI group). The acupoints of "Sanyinjiao" (SP 6), "Housanli" (ST 36), "Zhaohai" (KI 6), "Huantiao" (GB 30) and "Yanglingquan" (GB 34) were selected for API group, and five non-acupoints were selected on gastrocnemius and adductor of ischemic hind limb for TMI group. Both groups were accepted BM-MSCs transplantion. Model rat with hind limb ischemia was established with the method of blocking the femoral artery and its branches. The changes of blood flow (perfuse unit, PU) was monitored with laser Doppler flowmetry (LDF). In order to describe the visual changes in blood flow, the PU index (PUI) was determined as the ratio of ischemic to non-ischemic hind limb blood perfusion. And also, the levels of VEGF,bFGF in serum were tested to analyze the immunohistochemical expression quantity of VEGF and bFGF.

RESULTSComparing with the model and the TMI groups, the PUI value on 3rd, 14th and 21th days after BM-MSCs transplantation were significantly increased in the API group (P < 0.05, P < 0.01). In contrast to the model group, the VEGF,bFGF levels in serum and the immunohistochemical expression quantity of VEGF and bFGF in the API and TMI groups were significantly increased (all P < 0.01).

CONCLUSIONTransplantation of BM-MSCs through the acupoint can more significantly and quickly increase the blood flow and cause the greater improvement on hind limb ischemia than that of through the way of muscle injection.

Acupuncture Points ; Animals ; Bone Marrow Transplantation ; Disease Models, Animal ; Female ; Humans ; Ischemia ; physiopathology ; therapy ; Lower Extremity ; blood supply ; Male ; Mesenchymal Stem Cell Transplantation ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Regional Blood Flow

This study was purposed to directly induce murine embryonic stem cells (ESC) into hematopoietic stem cells (HSC) by simulating the spatial and temporal hematopoietic microenvironment changes in embryonic development, and to investigate the function of in vivo hematopoietic reconstitution of these HSC. E14 ESC were induced into embryoid body (EB) firstly. Then the cells from EB were further co-cultured with human aorta-gonad-mesonephros (AGM) region, fetal liver (FL) and bone marrow (BM) stromal cells in Transwell non-contact system in sequential orders. After 6 days of each co-cultured stage, the induced cells derived from EB were collected to analyze the Sca-1(+)c-Kit(+) cells by flow cytometry, check teratoma formation and transplant to BALB/C female mice exposed to lethal dose (60)Co γ-ray. The recipient mice were divided randomly into 5 groups: AGM, AGM + FL, AGM + FL + BM, irradiation control and normal control groups. The survival rates, hematopoietic reconstitution and engraftment of donor cells in the different groups were monitored. The results showed that Sca-1(+)c-Kit(+) cell level in EB cells co-cultured with human AGM region and FL stromal cells reached to peak value (21.96 ± 2.54)%. Teratomas could be found in NOD-SCID mice after subcutaneous injection of EB cells co-cultured with human AGM region stromal cells, while there was no teratoma in the mice after subcutaneous injection of EB cells induced by human AGM region and FL stromal cells. The recipients in AGM group and irradiation control group all died. The survival rate was 77.8% in AGM+FL group, and 66.7% in AGM+FL+BM group. The peripheral blood cell count was near normal at day 21 after transplantation, and Sry gene copies from donor could be detected in recipient mice of these two groups. It is concluded that sequentially inductive system with feeder cells from human AGM region, fetal liver and bone marrow simulating embryonic defined hematopoiesis procedures can enhance the directed differentiation of ESC into HSC which can safely reconstitute hematopoiesis in vivo.
Animals ; Aorta ; Cell Differentiation ; Cells, Cultured ; Coculture Techniques ; Embryonic Stem Cells ; cytology ; Female ; Hematopoiesis ; Hematopoietic Stem Cells ; cytology ; Humans ; Mesonephros ; Mice ; Mice, Inbred BALB C ; Mice, SCID

This study was aimed to investigate the effect of human aorta-gonad-mesonephros (AGM) region stromal cells on differentiation of murine embryonic stem cells (ESC) into hematopoietic stem cells (HSC) in vitro and to clarify their effect mechanism. E14 murine ESC were induced into embryo body (EB) firstly. Then the EB cells were further co-cultured with the stromal cells from human AGM region, fetal liver (FL) or bone marrow (BM) in Transwell non-contact system. According to the different culture methods, the EB cells were divided into 6 groups including EB control group, AGM group, FL group, BM group, AGM + FL group and AGM + BM group. The induced cells derived from EB were collected for Sca-1(+)/c-Kit(+) cells analysis by flow cytometry and colony forming unit (CFU) assay. The results showed that Sca-1(+)/c-Kit(+) cell proportion of EB cells significantly increased after being induced by different stromal cells (p < 0.01). The AGM + FL group had most Sca-1(+)/c-Kit(+) cells for the positive cell proportion reached (21.96 ± 2.54) % (p < 0.01). The Sca-1(+)/c-Kit(+) cell proportion of AGM + BM group was much high than that of BM group too (p < 0.01). The EB control group showed CFU amount less than any other groups, while the CFU amount of AGM + FL, AGM + BM groups were higher, especially in the AGM + FL group (p < 0.01). It is concluded that the human AGM region stromal cells may help to maintain certain number of primitive HSC with high proliferation potential. Human AGM region, FL or BM stromal cells, applied in sequential orders, can significantly expand in vitro the primitive hematopoietic stem/progenitor cells derived from ESC.
Animals ; Cell Differentiation ; Cell Line ; Coculture Techniques ; Embryonic Stem Cells ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Mesonephros ; cytology ; Mice ; Stromal Cells ; cytology

OBJECTIVETo investigate the biomechanical effect of different volume, distribution and leakage to adjacent disc of bone cement on the adjacent vertebral body by three-dimensional osteoporosis finite element model of lumbar.

METHODSL(4)-L(5) motion segment data of the cadaver of an old man who had no abnormal findings on roentgenograms were obtained from computed tomography (CT) scans. Three-dimensional model of L(4)-L(5) was established with Mimics software, and finite element model of L(4)-L(5) functional spinal unit (FSU) was established by Ansys 7.0 software. The effect of different loading conditions and distribution of bone cement after vertebroplasty on the adjacent vertebral body was investigated.

RESULTSThis study presented a validated finite element model of L(4)-L(5) FSU with a simulated vertebroplasty augmentation to predict stresses and strains of adjacent untreated vertebral bodies. The findings from this FSU study suggested the endplate and disc stress of the adjacent vertebral body was not influenced by filling volume of bone cement but unipedicle injection and leakage to the disc of bone cement could concentrate the stress of adjacent endplate.

CONCLUSIONSAsymmetric distributions and leakage of cement into intervertebral disc can improve the stress of endplate in adjacent vertebral body. These results suggest that optimal biomechanical configuration should have symmetric placement and avoid leakage of cement in operation.

Bone Cements ; adverse effects ; pharmacology ; Finite Element Analysis ; Humans ; Imaging, Three-Dimensional ; Lumbar Vertebrae ; surgery ; Male ; Polymethyl Methacrylate ; adverse effects ; pharmacology ; Stress, Mechanical

OBJECTIVEto compare the therapeutic effect of posterolateral fusion (PLF), posterior lumbar interbody fusion (PLIF) and posterior circumferential fusion (PCF) for lumbar spondylolisthesis.

METHODSfrom January 2003 to December 2008, 232 patients with lumbar spondylolisthesis treated with pedicle screw fixation and followed for reviewed retrospectively. The patients were divided into three groups based on fusion method: group A (PLF, 66 case), group B (PLIF, 54 case)and group C (PCF, 112 case). The three groups were reviewed and compared for clinical outcome and fusion rate.

RESULTSthe mean follow-up period was 21 months (range, 6-60 months). The fusion rate was 80.1% for PLF, 92.5% for PLIF and 93.7% for PCF group (P > 0.05). As to isthmic spondylolisthesis or Meyerding grade degenerative II and III spondylolisthesis, the fusion rate was 60.7% for PLF group, 90% for PLIF group and 93.3% for PCF group (P < 0.05). Compare the fusion rate for PLF group and PLIF+ PCF group (P < 0.05), fusion rate for PLIF group and PCF group (P > 0.05). The rate of excellent and good together was 84.8% in PLF group, 90.7% in PLIF group and 93.6% in PCF group (P > 0.05).

CONCLUSIONSposterior lumbar interbody fusion and posterior circumferential fusion are more consistent with bio-mechanics, have a higher fusion rate, for the treatment of spondylolisthesis they are the preferred surgical approaches.

Adolescent ; Adult ; Aged ; Bone Transplantation ; methods ; Female ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fusion ; methods ; Spondylolisthesis ; surgery ; Treatment Outcome ; Young Adult

Objective To investigate the epidemiological characteristics of shoulder injuries by training,and provide scientific evidence for effective intervention measures.Methods Through cluster random sampling method,a questionnaire survey was conducted on 1 080 trainers to analyze the occurrence of shoulder injury by training,the training courses lead to shoulder injuries and classification of shoulder injuries by training.The psychological status,shoulder symptoms,protective measures and other factors were compared between the injured group and the normal group to explore the influencing factors of injuries.Results Of 1 080 trainers,the incidence of shoulder injuries was 18.52％.The training course where injured were currently creeping forward (34.96％),push-ups (13.82％) and pull-ups (13.41％).The main types of injuries were soft tissue injuries(92.27％).The difference was statistically significant between two groups in the aspects of the mental state,shoulder protection measures and the presence of symptoms (P ＜ 0.01).Conclusion The shoulder injuries of training personnel were mainly caused by soft tissue injuries.Long time training,short time for rest,lack of control or confidence were the major risk factors.Shoulder symptoms during training and inadequate application of protective measures may lead to the incidence of training injuries increasing.

The climate on the plateau is characterized by hypobaria and hypoxia,where the natural environment is harsh,which poses a serious threat to the health and combativeness of troops.As a vital organ of the human body,the kidney has a complex vascular structure,large oxygen consumption,and is vulnerable to damage from hypoxia and other factors.This paper outlines the special changes of renal function and current diagnosis and treatment of kidney injury by taking into consideration the actual conditions on the plateau and renal blood and oxygen supply.Meanwhile,related countermeasures are recommended to facilitate the prevention and treatment of kidney injury in warfare on the plateau.

OBJECTIVETo discuss the minimal invasive arthroscopic surgery technique and clinical results of both the medial and lateral meniscal transplantation following the anterior cruciate ligament reconstruction with double bundles and bone tunnels.

METHODSIn August 2008 a minimal invasive surgery of both the medial and lateral meniscal allograft transplantation following anterior cruciate ligament reconstruction was preformed for 1 case with both the medial and lateral meniscectomy by arthroscopic surgery. The method of two bone plugs attached on tibial plateau was employed for medial meniscal allograft transplantation and the technique the bridge in slot for lateral meniscal allograft transplantation. The VAS, Lysholm score and IKDC rating were recorded before and after operation. The stability of knee was assessed by Lachman test, drawer sign and pivot shift test.

RESULTSThe patient was followed up 26 month after the operations. The degrees of knee flexion, extension and function of walk were normal. The Lachman test, drawer sign and pivot shift test were nearly normal. The VAS after operation was 2 points lower than that before operation. The Lysholm score post-operation was 20 points higher than pre-operation. The IKDC became B degree in late following-up from C degree before the operation. MRI revealed anterior cruciate ligament graft was continuous and the meniscal allograft was normal shape on year 1 after the operation. The posterior horn of medial meniscal allograft and anterior corner of lateral meniscal allograft showed slightly shrunk. The second-look arthroscopy showed that the healing occurring between meniscal allograft and the capsule and meniscal allograft was normal shape on month 18 after the operation. The anterior horn of medial and lateral meniscus was slightly worn.

CONCLUSIONSBoth the medial and lateral meniscal transplantation following the anterior cruciate ligament reconstruction in appropriately selected patients with the medial and lateral meniscus-deficient knee may recover the knee mechanic balance and stability, which is a option of treatment for that young and activity patients. It is proposed that the medial and lateral meniscal grafts harvested from a single donator. Attention should be paid to the direction of the bone tunnels fixing the horns of the meniscus in order to avoid communication with the tunnels of anterior cruciate ligament reconstruction.

Anterior Cruciate Ligament ; surgery ; Anterior Cruciate Ligament Reconstruction ; methods ; Arthroscopy ; Humans ; Knee Injuries ; surgery ; Male ; Menisci, Tibial ; transplantation ; Transplantation, Homologous ; Treatment Outcome ; Young Adult

OBJECTIVETo establish a flow cytometric method to detect the alteration of phenotypes and concentration of circulating microvesicles (MVs) from myocardial ischemic preconditioning (IPC) treated rats (IPC-MVs), and to investigate the effects of IPC-MVs on ischemia/reperfusion (I/R) injury in rats.

METHODSMyocardial IPC was elicited by three.cycles of 5-min ischemia and 5-min reperfusion of the left anterior descending (LAD) coronary artery. Platelet-free plasma (PFP) was isolated through two steps of centrifugation at room temperature from the peripheral blood, and IPC-MVs were isolated by ultracentrifugation from PFR PFP was incubated with anti-CD61, anti-CD144, anti-CD45 and anti-Erythroid Cells, and added 1, 2 µm latex beads to calibrate and absolutely count by flow cytometry. For functional research, I/R injury was induced by 30-min ischemia and 120-min reperfusion of LAD. IPC-MVs 7 mg/kg were infused via the femoral vein in myocardial I/R injured rats. Mean arterial blood pressure (MAP), heart rate (HR) and ST-segment of electro-cardiogram (ECG) were monitored throughout the experiment. Changes of myocardial morphology were observed after hematoxylin-eosin (HE) staining. The activity of plasma lactate dehydrogenase (LDH) was tested by Microplate Reader. Myocardial infarct size was measured by TTC staining.

RESULTSTotal IPC-MVs and different phenotypes, including platelet-derived MVs (PMVs), endothelial cell-derived MVs (EMVs), leucocyte-derived MVs (LMVs) and erythrocyte-derived MVs (RMVs) were all isolated which were identified membrane vesicles (<1 Vm) with corresponding antibody positive. The numbers of PMVs, EMVs and RMVs were significantly increased in circulation of IPC treated rats (P<0.05, respectively). In addition, at the end of 120-min reperfusion in I/R injured rats, IPC-MVs markedly increased HR (P<0.01), decreased ST-segment and LDH activity (P < 0.05, P < 0.01). The damage of myocardium was obviously alleviated and myocardial infarct size was significantly lowered after IPC-MVs treatment (P < 0.01).

CONCLUSIONThe method of flow cytometry was successfully established to detect the phenotypes and concentration alteration of IPC-MVs, including PMVs, EMVs, LMVs and RMVs. Furthermore, circulating IPC-MVs protected myocardium against I/R injury in rats.

Animals ; Cell-Derived Microparticles ; metabolism ; Coronary Vessels ; pathology ; Flow Cytometry ; Heart Rate ; Ischemic Preconditioning, Myocardial ; Myocardial Infarction ; physiopathology ; Myocardial Reperfusion Injury ; physiopathology ; Myocardium ; pathology ; Phenotype ; Rats