Objective The purpose of the present study was to observe the muscle glucose metabolism by determining the changes in sarcolemma GLUT4 protein content and muscle AMPK activity in overtraining rats.Methods Twenty-seven SD rats were randomly divided into group A(sedentary),group B(underwent endurance exercise 60 minutes/day)and group C(underwent endurance exercise 120 minutes/day).Both exercise groups trained 6 days/wk for 9 weeks.Muscle glycogen,sarcolemma GLUT4 contents and muscle AMPK activity were determined after the 9-week training.Results Sarcolemma GLUT4 content and muscle AMPK activity increased,muscle glycogen contents tended to rise in group B as compared with that in group A.Sarcolemma GLUT4 content and muscle AMPK activity decreased,but muscle glycogen contents were equivalent in group C as compared with that in group B.Conclusion Muscle AMPK activity deceased,and the translocation of GLUT4 protein was inhibited in the overtraining rats.The results could not support a glycogen depletion hypothesis.

Objective To investigate the technical feasibility of medical robot in application to vascular intervention. Methods The independent-developed medical robot was used in the glass vessel model and vascular intervention experiments in a dog. Results The process of experiments were smooth,the system movement did not have any malfunction,and the animal experiments did not have any operative complications. The operative time was 50 minutes.Conclusions The medical robot can basically meet the requirements of cerebral angiography. It has laid a foundation for further development of intracranial vascular interventional procedures and clinical application.

Pigs are considered as ideal donors for xenotransplantation because they have many physiological and anatomical characteristics similar to human beings. However, antibody-mediated immunity, which includes both natural and induced antibody responses, is a major challenge for the success of pig-to-primate xenotransplantation. Various genetic modification methods help to tailor pigs to be appropriate donors for xenotransplantation. In this study, we applied transcription activator-like effector nuclease (TALEN) to knock out the porcine α-1, 3-galactosyltransferase gene GGTA1, which encodes Gal epitopes that induce hyperacute immune rejection in pig-to-human xenotransplantation. Meanwhile, human leukocyte antigen-G5 gene HLA-G5, which acts as an immunosuppressive factor, was co-transfected with TALEN into porcine fetal fibroblasts. The cell colonies of GGTA1 biallelic knockout with positive transgene for HLA-G5 were chosen as nuclear donors to generate genetic modified piglets through a single round of somatic cell nuclear transfer. As a result, we successfully obtained 20 modified piglets that were positive for GGTA1 knockout (GTKO) and half of them expressed the HLA-G5 protein. Gal epitopes on the cell membrane of GTKO/HLA-G5 piglets were completely absent. Western blotting and immunofluorescence showed that HLA-G5 was expressed in the modified piglets. Functionally, the fibroblasts from the GTKO/HLA-G5 piglets showed enhanced resistance to complement-mediated lysis ability compared with those from GTKO-only or wild-type pigs. These results indicate that the GTKO/HLA-G5 pigs could be a valuable donor model to facilitate laboratory studies and clinics for xenotransplantation.
Animals ; Animals, Genetically Modified ; Gene Knockout Techniques ; HLA Antigens ; Humans ; Nuclear Transfer Techniques ; Swine ; Transplantation, Heterologous

Objective The finite element method was used to analyze the effect of height of bone graft on adjacent segment stress after anterior cervical decompression and fusion.Methods One healthy adult male was included in this study.A detailed,geometrically accurate three-dimensional cervical spine model had been successfully developed from computed tomography (CT) scanning of a healthy adult male.A pure moment of 1.0 N· m was applied in different directions on the upper endplate of C3 to simulate flexion,extension,left-right lateral bending,and left-right axial rotation.We did biomechanical experiment with goat cervical specimens in terms of the moment-rotation responses.The finite element (FE) model was compared and validated with the experimental data.The relevant data in the literature was also applied for comparison and validation.To construct the post-surgery model,we turned the property of disk annulus and disk nucleus into that of the cancellous bone at C4-5.The internal fixation systems were also implanted.We changed the height of bone graft in C4-5 to be 90％,150％,175％ and 200％ of the height between C4-5 pre-surgery.The finite element models were built to simulate the post-surgery scenarios with different bone graft height,respectively.A stress analysis was conducted on the bone graft and adjacent disks.Results Under different height of bone graft,the maximum yon Mises stress on C3-4 showed when the height of bone graft was 200％.The values were 0.99 MPa,0.85 MPa,0.91 MPa and 0.89 MPa in different loading condition.The values at 150％ of the height between C4-5 pre-surgery were only 81％,73％,80％ and 87％ of the values at 200％.For C5-6,the maximum yon Mises stress were 0.77 MPa,0.83 MPa,0.91 MPa and 0.81 MPa,observed at the height of bone graft was 175％ except extension.The values at 150％ of the height between C4-5 pre-surgery were only 76％,93％,86％ and 78％ of the biggest values.With regard to C4-5,the biggest yon Mises stress was 1.25 MPa,1.77 MPa,1.75 MPa and 1.81 MPa.It was observed when the height of bone graft was 200％ of the height between C4-5 pre-surgery.The values were smallest under height of 150％ of the height between C4-5 pre-surgery.The smallest values were 63％,63％,75％ and 52％ of the biggest values.The higher the bone graft was,the bigger the maximum von Mises stress on bone graft.In a word,it seems that the smallest stress was observed when the bone graft height was 150％.Concltusion The height of bone graft makes an important difference on the stress of adjacent segment after anterior cervical decompression and fusion.150％ of the height between C4-5 presurgery was proper bone graft height.In this case,the stress on the adjacent segments intervertebral disc is smaller.The probability of adjacent segments degeneration is smaller.

OBJECTIVETo verify the feasibility and safety of the vascular interventional vascular interventional surgical robot system applied to vascular interventional operation.

METHODSFrom March to September 2013, 10 patients had undergone robot-assisted cerebral angiography. There were 6 male and 4 female patients; aged from 19 to 58 years, with an average age of 38.4 years. The operation were carried out by neurosurgeons and vascular interventional robot. After successfully implanted of femoral artery sheath by hand, the catheter was fixed on the robot, under the guidance of navigation image the surgeon manipulate the master part and control the slave part of robot by sending command through network transmission, finally finished the whole cerebral angiography. The operation time was recorded from placing the sheath into femoral artery to finishing cerebrovascular selective angiography, simultaneously the time of staff under exposure of X ray was recorded, and the position difference between the setted targets and the actual position(positioning accuracy).

RESULTSIt took 25-41 minutes to finish the cerebral angiography, the average time was (31 ± 5) minutes, and the robot-assisted angiography went quickly and smoothly without surgical complications. The remote positioning accuracy was (1.03 ± 0.23) mm. The time of staff under exposure of X ray was 0 minute, the entire experimental process was basically implemented mechanization and automation.

CONCLUSIONThis system basically achieves initial medical purposes, such as reducing the radiation, facilitating interventional procedures on the basis of enhancing the image navigation, shorting the operation time, and improve the quality of operation.

Adult ; Female ; Humans ; Male ; Middle Aged ; Robotic Surgical Procedures ; Vascular Surgical Procedures ; instrumentation ; Young Adult

OBJECTIVETo explore the methods and results of surgical management for refractory dysphagia and aspiration.

METHODSThe clinical data of 24 refractory dysphagia and aspiration patients who accepted surgical management were retrospectively analysed.

RESULTSTwenty-four refractory dysphagia and aspiration patients accepted 26 operations between 2001 and 2014. Of the 26 operations, 17 were cricopharyngeal myectomy (CPM), 6 were scarectomy, 3 were laryngeal-tracheal separation. No severe complications occurred. Assessments of dysphagia were completed in 18 operations before and after operation. Aspiration scores of videofluoroscopic swallowing study (VFSS) were 4.50 [4.00;7.00] vs 2.00 [1.00; 3.25], P = 0.000; swallow dysfunction scroes of VFSS were 5.00 [4.00; 12.00] vs 1.00 [1.50; 10.00], P = 0.001; aspiration scores of fibroptic endoscopic evaluation of swallowing (FEES) were 4.00 [5.00; 7.00] vs 2.00 [1.75; 3.00], P = 0.000. But the surgical results for post radiotherapy dysphagia were not successful (n = 5): aspiration scores of VFSS were 7.00 [6.50; 8.00] vs 6.00 [2.00; 7.50], P = 0.109;swallow dysfunction scroes of VFSS were 12.00 [10.50; 12.00] vs 12.00 [7.50; 12.00], P = 0.180;aspiration scores of FEES were 7.00 [6.50; 8.00] vs 6.00 [2.00; 7.50], P = 0.109.

CONCLUSIONSurgical management was effective for refractory dysphagia and aspiration, but the surgical indication selection should be strict.

Deglutition Disorders ; surgery ; Endoscopy ; Fluoroscopy ; Humans ; Larynx ; Larynx, Artificial ; Retrospective Studies ; Trachea

OBJECTIVETo explore the effects of Yishen Zhuanggu decoction ([Chinese characters: see text], YSZGD) on the fracture healing and function of wrist joint in osteoporotic distal radius fractures (ODRF) of elderly patients.

METHODSNinety patients of ODRF were treated with splint external fixation and YSZGD, including 28 males and 62 females, with an average age of 63 years (ranged, 61 to 91 years). According to AO classification, all the patients were classified as type A or B1. Among the patients, 80 patients had a type of Colles fracture,7 patients were with a type of Smith fracture,and 3 patients had a type of Barton fracture. They were randomly divided into three groups included TCM, Western medicine and control group (30 cases each group) after the fractures were fixed manually by splint. Patients in TCM group took YSZGD orally, one potion and two time each day; the patients in Western medicine group treated with Calcitonin (50 units, intramuscular, 2 times each week), Caltrate D (1 tablets, taken orally, one time each day) and Alfacalcidol Soft Capsules (0.5 units, taken orally, one time each day); and the patients in control group took no medicine. X-ray examination on the fractured wrist was taken at the 4, 6, 8, 12th weeks after treating to observe, the fracture healings, the evaluation of wrist function and comparision of curative effects were taken at the 2nd month.

RESULTSThe fracture healing time in TCM group was 7.12 +/- 2.32 weeks and that in the Western medicine group was 9.25 +/- 3.05 weeks, showing significant differences between them (PC < 0.05), and that in the control group was 11.57 +/- 1.93 weeks which was longer thant in that in the two medicine groups (P < 0.05); According to Dienst wrist rating system, the excellent and good rates of three groups were 93.3%, 86.7% and 60% respectively which showing significant differences between medicine groups and control group (P < 0.05), and the curative effects of medicine groups were more excellent than that of control group through Ridit analysis.

CONCLUSIONYSZGD can promote the fracture healing evidently and shorten the healing time, whose curative effect surpassed Western medicine and promoted the restoration of wrist function.

Aged ; Aged, 80 and over ; Bone Density Conservation Agents ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Fracture Fixation ; Humans ; Male ; Radius ; surgery ; Radius Fractures ; drug therapy ; surgery ; Treatment Outcome

OBJECTIVETo evaluate the effect of reconstruction of anterior cruciate ligaments (ACL) with allogeneic tendon or autologous hamstring tendon under arthroscopy.

METHODSThirty-two cases of ACL injury (3 cases compound with PCL injury) were reviewed. All the patients were divided into two groups randomly. Fifteen cases were reconstructed with hamstring tendon, including 12 male and 3 female with the age ranging from 23 to 61 years. Seventeen cases were reconstructed with allograft, including 11 male and 6 female with the age ranging from 17 to 57 years. The tendons were fixed with absorbable or Ti screws. All patients were recorded symptoms,physical signs and Lysholm Scores two weeks after operation,and functional rehabilitation of knee six months after operation.

RESULTSAll cases were followed up from 6 to 8 months. Pain and clinical symptoms disappeared and the function of knee joint was improved in all patients. Five patients of allograft with positive Lachman test result,one with severe graft rejection was found complete absorption of allograft under arthroscopy. No significant differences were seen in Lysholm Score between the two groups,average (88.5 +/- 7.2), (93.2 +/- 8.5) after operation (P > 0.05).

CONCLUSIONThe effects of reconstruction of cruciate ligaments with allogeneic tendon or autologous hamstring tendon under arthroscopy are more satisfying. But there are more symptoms and obvious individual differences in the earlier stage of rehabilitation. So reconstruction of cruciate ligaments under arthroscopy using autologous hamstring tendon should be performed as far as possible.

Adolescent ; Adult ; Anterior Cruciate Ligament ; surgery ; Arthroscopy ; Female ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Tendons ; transplantation ; Transplantation, Homologous

Human induced pluripotent stem cells (hiPSCs) are promising in regenerative medicine. However, the pluripotent stem cells (PSCs) may form clumps of cancerous tissue, which is a major safety concern in PSCs therapies. Rapamycin is a safe and widely used immunosuppressive pharmaceutical that acts through heterodimerization of the FKBP12 and FRB fragment. Here, we aimed to insert a rapamycin inducible caspase 9 (riC9) gene in a safe harbor AAVS1 site to safeguard hiPSCs therapy by drug induced homodimerization. The donor vector containing an EF1α promoter, a FRB-FKBP-Caspase 9 (CARD domain) fusion protein and a puromycin resistant gene was constructed and co-transfected with sgRNA/Cas9 vector into hiPSCs. After one to two weeks screening with puromycin, single clones were collected for genotype and phenotype analysis. Finally, rapamycin was used to induce the homodimerization of caspase 9 to activate the apoptosis of the engineered cells. After transfection of hiPSCs followed by puromycin screening, five cell clones were collected. Genome amplification and sequencing showed that the donor DNA has been precisely knocked out at the endogenous AAVS1 site. The engineered hiPSCs showed normal pluripotency and proliferative capacity. Rapamycin induced caspase 9 activation, which led to the apoptosis of all engineered hiPSCs and its differentiated cells with different sensitivity to drugs. In conclusion, we generated a rapamycin-controllable hiPSCs survival by homodimerization of caspase 9 to turn on cell apoptosis. It provides a new strategy to guarantee the safety of the hiPSCs therapy.
Humans ; Induced Pluripotent Stem Cells ; Sirolimus/metabolism* ; Caspase 9/metabolism* ; RNA, Guide, CRISPR-Cas Systems ; Pluripotent Stem Cells/metabolism* ; Cell Differentiation ; Puromycin/metabolism*

Nine new compounds, including five natural rarely-occurring 2, 3-dihydro-1H-indene derivatives named diaporindenes E-I (1-5), and four new benzophenone analogues named tenellones J-M (6-9) were isolated from the deep-sea sediment-derived fungus Phomopsis lithocarpus FS508. All the structures for these new compounds were fully characterized on the basis of spectroscopic data, NMR spectra, and ECD calculation and single-crystal X-ray diffraction analysis. The potential anti-tumor activities of compounds 1-9 against four tumor cell lines SF-268, MCF-7, HepG-2, and A549 were evaluated using the SRB method. Compound 7 exhibited cytotoxic activity against the SF-268 cell line with an IC
Antineoplastic Agents/pharmacology* ; Cell Line, Tumor ; Crystallography, X-Ray ; Fungi ; Molecular Structure ; Phomopsis