ObjectiveTo observe the effect of ginsenoside Rg₁ （G-Rg₁） on the biological activity of cryopreserved Schwann cells （SCs） of the rat sciatic nerve and explore the feasibility of G-Rg₁ in reducing the cryopreservation-induced injury in SCs. MethodBilateral sciatic nerves of SD rats were randomly divided into a fresh group， a blank group， and five G-Rg₁ groups of different doses （1×10^-7， 1×10^-6， 1×10^-5， 1×10^-4， and 1×10^-3 mol·L^-1）. The nerves in the blank group and the G-Rg₁ groups were preserved in liquid nitrogen solutions containing 0， 1×10^-7， 1×10^-6， 1×10^-5， 1×10^-4， and 1×10^-3 mol·L^-1 G-Rg₁ for four weeks. The apoptosis of SCs was detected by TdT-mediated dUTP-biotin nick end labeling （TUNEL）/S100 immunofluorescence staining. The expression of cysteinyl aspartate-specific protease （Caspase）-9， Caspase-3， major histocompatibility complex （MHC）-Ⅰ， and MHC-Ⅱ was detected by Western blot. Subsequently， all nerves were cultured in the incubator at 37 ℃ with 5% CO₂ for 7 days. The expression of glial cell line-derived neurotrophic factor （GDNF） and nerve growth factor （NGF） was detected by Western blot. In addition， the above cryopreserved nerves in the blank group and the 1×10^-6， 1×10^-5， and 1×10^-4 mol·L^-1 G-Rg₁ groups were transplanted to the Wistar rats by allografting （blank transplantation group and the 1×10^-6， 1×10^-5， and 1×10^-4 mol·L^-1 G-Rg₁ transplantation groups）， and fresh sciatic nerve allograft and isograft control group were set up. Sixteen weeks after transplantation， compound muscle action potential （CMAP） and nerve conduction velocity （NCV） were measured by electrophysiology. Nerve filament （NF）200 immunofluorescence staining， transmission electron microscopy， and toluidine blue staining were used to analyze the histology of the regenerated nerves. ResultCompared with the fresh group， the blank group and the G-Rg₁ groups showed increased expression of Caspase-9， Caspase-3， and the apoptosis of SCs （P<0.05，P<0.01） and decreased expression of GDNF， NGF， MHC-Ⅰ， and MHC-Ⅱ （P<0.01）. Compared with the results in the blank group， the expression of Caspase-9 and Caspase-3 decreased in the 1×10^-7， 1×10^-6， 1×10^-5，1×10^-4 mol·L^-1 G-Rg₁ groups （P<0.01）， and the apoptosis of SCs was reduced in the 1×10^-7-1×10^-4 mol·L^-1 G-Rg₁ groups（P<0.05，P<0.01） and increased in the 1×10^-3 mol·L^-1 group （P<0.05）， while the expression of GDNF and NGF increased in the 1×10^-7， 1×10^-6， 1×10^-5，1×10^-4 mol·L^-1 G-Rg₁ groups and decreased in the 1×10^-3 mol·L^-1 group （P<0.05）. There was no statistical significance in the expression of MHC-Ⅰ and MHC-Ⅱ between the blank group and the G-Rg₁ groups. Compared with the 1×10^-7 mol·L^-1 and 1×10^-3 mol·L^-1 G-Rg₁ groups， the 1×10^-6 1×10^-5， 1×10^-4 mol·L^-1 G-Rg₁ groups showed decreased expression of Caspase-3 and the apoptosis of SCs （P<0.05，P<0.01） and increased expression of GDNF and NGF （P<0.05，P<0.01）. There was no statistical significance in MHC-Ⅰ and MHC-Ⅱ expression among G-Rg₁ groups. Sixteen weeks after transplantation， compared with the isograft group， the blank transplantation group and the G-Rg₁ transplantation groups showed decreased CMAP， NCV， myelin sheath thickness， and number of myelinated nerve fibers （P<0.01）， and the 1×10^-6 and 1×10^-4 mol·L^-1 G-Rg₁ transplantation groups showed decreased NF200 （P<0.01）. Compared with the allograft group， the blank transplantation group and the G-Rg₁ transplantation groups showed increased CMAP， NCV， NF200， myelin sheath thickness， and number of myelinated nerve fibers （P<0.05，P<0.01）. Compared with the blank transplantation group， the G-Rg₁ transplantation groups showed increased CMAP， NCV， NF200， myelin sheath thickness， and number of myelinated nerve fibers （P<0.05，P<0.01）. Among all groups of G-Rg₁ transplantation， each index of the 1×10^-5 mol·L^-1 G-Rg₁ transplantation group was superior to that of the 1×10^-4 and 1×10^-6 mol·L^-1 G-Rg₁ transplantation group （P<0.05）. ConclusionG-Rg₁ at a certain centration can maintain the biological activity of cryopreserved SCs of rat sciatic nerve， alleviate the cryopreservation-induced injury of rat sciatic nerve， and promote nerve regeneration after allograft.

Objective To investigate the effects of triptolide (T10) on biological activity of sciatic nerve in cold preservation and nerve regeneration after allogeneic transplantation. Methods Cell Counting Kit-8 (CCK-8) was used to test the proliferation of SCs in logarithmic phase in 1×10-6 mol/L, 1×10-7 mol/L, 1×10-8 mol/L and 1×10-9 mol/L of T10 solution. The sciatic nerves from Sprague-Dawley rats were pretreated in 0 mol/L, 1×10-6 mol/L, 1×10-7 mol/L, 1×10-8 mol/L and 1×10-9 mol/L of T10 solution at 4 ℃ or 37 ℃ for 24 hours (n = 6). The expression of nerve growth factor (NGF), glial cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) was detected with Western blotting. Other sciatic nerve fragments were randomly divided into fresh nerve group (group A, n = 30), DMEM preservation group (group B, n = 30), T10 preservation group (group C, n = 30), T10 pretreatment DMEM preservation group (group D, n = 30) and T10 pretreatment T10 preservation (group E, n = 30), and were stored under 4 ℃ for four weeks. Calcein-AM/PI double staining laser confocal microscope and flow cytometry were used to detect the living cells and dead cells. The expression of the major histocompatibility complex (MHC)-I, MHC-II and intercellular cell adhesion molecule-1 (ICAM-1) was detected with Western blotting. The corresponding sciatic nerves were used to repaire 10 mm defects in Wistar rats (named groups A', B', C', D' and E'), and fresh sciatic nerve from Wistar rats were also used to do it (group F'). Compound muscle action potential (CMAP) and motor nerve conduction velocity (MNCV) were tested 16 weeks after transplantation, and then the grafts were observed for the nerve regeneration. Results SCs proliferated as the controls in the T10 solution with a concentration of 1×10-9 to 1×10-7 mol/L (P > 0.05). The expression of all the neurotrophic factors was more under 37 ℃ than under 4 ℃ in all the concentrations of T10 solution, and it was the most in the concentration of 1×10-8 mol/L whenever under 37 ℃ or 4 ℃ (P < 0.05). After four weeks of cold preservation, compared with groups B, C and D, the living nerve cells were the most in group E, and the expression of MHC-I, MHC-II and ICAM-1 was the least (P < 0.05). CMAP, MNCV and the never regeneration were better in group E' than in groups A', B', C' and D' (P < 0.05). A large number of myelinated nerve fibers were observed in groups E' and F', uniformity in size, wide distribution, and with myelin sheath, compared with those in groups A', B', C' and D'. Conclusion A certain concentration of T10 can induce the sciatic nerve of rats to express neurotrophic factor in vitro, which can improve the biological activity of cold preservation nerves, reduce the immunogenicity, and promote the regeneration of recipient nerve after allogeneic transplantation. It is even better to be pretreated with T10 before cold preservation.

Objective To assess the factors associated with the restoration of spontaneous circulation (ROSC) and 2-year survival prognosis in patients with cardiac arrest (CA) after acute myocardial infarction (AMI),and after ROSC,the effects of various factors on midian survival time and on 2-year survival.Methods In a registry study from January 2005 to January 2015,all consecutive AMI-induced CA patients treated with cardiopulmonary resuscitation (CPR) admitted to our hospital were enrolled.The survivors were followed-up for 2 years.Univariate analysis was applied to evaluate factors associated with rate of ROSC and 2-year survival.Multivariable logistic regression analysis was applied to evaluate statistically significant factors in the univariate analysis.Medians with inter-quartile ranges were used to describe 2-year survival time affected by various factors after ROSC.Kaplan-Meier survival curve analysis was used to evaluate the effect of factors on 2-year survival.Results A total of 254 cases with CA after AMI were enrolled,including 129 cases of ROSC and 71 cases of 2-year survival.Univariate analysis showed age ≥ 70 years,CA occurred during 22:00-8:00,the duration time ofCPR ≥ 15 min and adrenaline dosage ＞ 5 mg were unfavorable predictors of ROSC;while,left ventricular ejection fraction (LVEF) ≥ 40％ before CA,shockable rhythm and percutaneous coronary intervention (PCI) therapy were favorable predictors.Besides,age ≥ 70 years,intubation during CPR,adrenaline dosage ＞ 5 mg and cardiogenic shock were unfavorable predictors of 2-year survival;While,male,normal daily activity before CA and PCI treatment were favorable predictors.Multivariable analysis showed age,the duration of CPR,adrenaline dosage,LVEF before CA,the rhythm during CPR and PCI therapy were independent predictors of ROSC.Age and PCI therapy were independent predictors of 2-year survival.Among patients,the survival time was affected by various factors after ROSC,and the factors with minimum 25％ and small median value were associated with cardiac rupture,cancer,adrenaline dosage ＞ 5 mg and cardiogenic shock.The factor with maximum 25％ value was PCI treatment (216 days).Kaplan-Meier survival analysis suggested that age ≥ 70 years was an unfavorable factor of 2-years survival (Log-rank test,P=0.007);while,PCI treatment was a favorable factor (Log-rank test,P＜0.01).PCI-related prognosis analysis showed that the effectiveness of PCI was related to the timing of PCI,the number of infarctrelated artery and the difference in culprit lesion.Conclusions The age ≥ 70 years was disadvantageous to both ROSC and 2-year survival.PCI treatment was favorable to both ROSC and 2-year survival.

Objective To investigate MRI manifestations of lumbar and proximal femoral bone marrow changes before and after recombinant human granulocyte colony stimulating factor (rhG-CSF) was subcutaneous injected for healthy adults.Methods Twenty healthy blood stem cell donors without hematologic disease were enrolled in this study. All of them underwent lumbar sagittal and proximal femur coronal MRI examination with spin echo T1 WI and fat-suppressed T2WI.The first examination were performed before subcutaneous injection of rhG-CSF for comparison. In 4-7 days and 30-60 days after injection, the other two examinations were performed. The signal changes of lumbar and proximal femoral bone marrow were investigated by reading pictures and calculating the contrasted noise ratio (CNR).ResultsBefore rhG-CSF injection, all patients presented normal signal intensity of hone marrow. In 4-7 days after injection, all the 20 cases presented homogeneous signal decrease in lumbar vertebral bodys on T1 WI, accompanied by reduced fatty signal. In proximal femur, patchy or stripped hypointensity areas were found in intertrochanteric and subtrochanteric areas on T1 WI. On fat-suppressed T2 WI images, the signal of lumbar and proximal femoral bone marrow changed to equal or slightly-high signal intensity. In all cases,abnormal signal areas presented in lumbar and proximal femoral bone marrow occurred simultaneously in the same case.In the 10 cases received the third MRI during 30-60 days after rhG-CSF injection, signal intensity of lumbar bone marrow turned to normal in all sequence, but abnormal signal intensity areas were still existed and extended to distal part in femoral bone marrow, which appeared as symmetric stripped or patchy equal or slightly-low signal intensity on T1 WI and equal or slightly-high signal intensity on T2 WI. The CNR of lumbar bone marrow to subcutaneous fat before rhG-CSF injection, in 4-7 days and 30-60 days after rhG-CSF injection were 114. 11 ± 15. 11,71.04 ± 12. 25 and 91.64 ± 1 I. 68, respectively. Significant difference was found between before rhG-CSF injection and 4-7 days after injection ( P ＜ 0. 05 ) , but no significant difference between the others( P ＞ 0. 05 ). Conclusion After injection of rhG-CSF, the short-term changes of hematopoietic cells and fat content in bone marrow can be displayed on MRI, which provided non-invasive information for bone marrow transplantation.

Objective To evaluate X-ray and MRI features of limbs in childhood acute leukemia.Methods Thirteen children with acute leukemia in our pediatric hematology ward were recruited.Allpatients were pathologically diagnosed by bone marrow aspiration and complained of bone or joint pain in the first visit.ConventionaI X-ray and MRI examinations of algesic sites were performed before clinical treatment and after complete remission.MR images were obtained with SE-T1WI,SE-T2WI and T2WI-fat suppressed sequences and symmetria bilateralis was requested while scanning.X-ray and MRI manifestations were evaluated and compared.Resuits All 13 patients had received X-ray examinations.Among them,6 had normal X-ray findings,whereas the other 7(14 sites)showed various abnormalities including radiolucent metaphyseal bands(5 sites),periosteal reaction(3 sites),osteapenia(2 sites),mixed lesions(lysissclerosis,1 site),and permeative pattern(3 sites).The number of patients for MRI examinations was 8(11 sites).Among them,6(9 sites)showed bone marrow infiluration and bone marrow necrosis accompanied by normal X-ray findings,another 2(2 sites)showed bone marrow infiltration associated with radiographic abnormalities of periosteal reaction and radiolucent metaphyseal bands.Four cases were followed up within 1 week when reached complete remission by chemotherapy.MR images features included reduced sizes of bone marrow infiltration lesions associated with increased signal intensity on T1WI,and disappearance of double-line sign on bone marrow necrosis accompanied by signal homogenization.However,the radiograph before and after treatment in the same cases did not differ significantly.Conclusions MRI was earlier and more comprehensive in showing limbs bone marrow abnormality than radiogram in acute leukemia children with chief complaint of osteoarticular pains.MRI might be one of indicators in following up therapeutic effect for AL children with osteoarticular disorder.

Objective To investigate the value of MRI and~1H-MRS in diagnosis of early stage of diabetic encephalopathy by detecting regional metabolite in cynomolgus diabetes models. Methods Five pathogen-free male adolescent cynomolgus were made type 1 diabetes mellitus models (T1DM) by intravenous injection of streptozotocin (STZ) (100 mg/kg), and the reliability and stability of the modes were assessed with long term follow-up of blood glucose and intravenous glucose tolerance tests. MRI and ~1H-MRS were performed to evaluate the volume, signal intensity and metabolic ratios of NAA/Cr, mI/Cr and Cho/Cr at hippocampus, lateral temporal lobe and occipital lobe 3 years after model establishment. Cortisol in serum was detected with immunoradiometric assay. In addition, 5 normal adult cynomolgus monkeys were selected in the control group and accepted the same examination above. Results ①Intravenous administration of STZ could made stable T1DM monkey model. ②Only mI/Cr ratio increased at hippocampus of diabetic monkeys compared to the control group (P<0.05). ③There was no statistical difference of cortisol in serum between the diabetic group and the control group (P＞0.05). Conclusion ~1H-MRS may detect the metabolic changes of the hippocampus in STZ-induced diabetic adolescent cynomolgus monkeys and may contributes to the early diagnosis of diabetic encephalopathy.

Objective:To improve diagnostic ability by the evaluation of imaging appearances of solitary plasmacytoma.Meth-ods:Seventeen patients with solitary plasmacytoma confirmed by biopsy or operation were included in our study,whose clinical features and diagnostic imaging appearances were included.Results:Single lesion was found in all cases;Lesions of vertebra in 5 cases,flat bone in 5 cases,pipe bone in 2 cases and outside the bone in 5 cases.The Lesions in bone showed expansive osteolytic with clear border,soft tissue mass nearby,and thin and broken cortical.Vertebra lesions showed lytic destruction and compressed fracture.MRI showed slightly hypointense in T1WI and hyperintense in T2WI with markedly homogeneous enhancement.Lesions outside the bone appeared soft tissue mass with similar density to muscle in CT and X ray,and isointen-sity in T1WI and T2WI with markedly homogeneous enhancement.Lesions in ECT showed medium-to-high nuclide concentration,and in PET/CT showed the increase of glucose metabolize,with SUV 6.3～8.6.About one month after effective chemical therapy,soft tissue mass shrank obviously and new bone appeared,with the decrease of glucose metabolize,and SUV under 2.0.Conclusion:Solitary plasmacytoma often occurs in male,and in younger age and lacks symptoms in the whole body compared with multiple myeloma.Lesions in different regions show different manifestations,but all borders are well defined.CT and MRI can depict the extent of solitary plasmacytoma of bone clearly,but MRI was better in showing bone marrow infiltration range,and showing compression of spinal cord.PET/CT are more suitable in follow-up therapy.

Objective To clone the cDNA encoding human HMGB1, express it in E. coli, and identify its biological activity. Methods Human HMGB1 cDNA was amplified by RT-PCR and cloned into vector pUC19. After sequence analysis, the cDNA was ligated into prokaryotic expression vector pQE-80L and induced by IPTG to express HMGB1. The protein was purified with Ni~(2+)-NTA chromatography and polymyxin B affinity column. To identify the function of purified protein, the product was co-cultured with THP1 cells. Results Recombinant expression plasmid pQE-80L/HMGB1 was constructed successfully. After purification, the protein purity reached 96%. The recombinant HMGB1 stimulated THP1 to secrete TNF-? . Conclusion The highly purified HMGB1 was obtained successfully, which showed biological activity. These results lay the foundation for further research on the function of human HMGB1.

Objective To find the X-ray characteristic and diadynamic method in congenital defect of muscular layer in stomach wall. Methods Six cases confirmed by operation and pathology were collected,underwent eight standing abdomen plain films,underwent 5 decubitus abdomen plain films in 6 cases. Results The appearances in standing plain film of freeing gas under diaphragm, eventration of diaphragm, passing through the whole abdomen of gas and liquid plane, vanished gastriole were found in 6 cases. Superior border of liver was found in 4 cases.The appearances in decubitus abdomen plain films of abdominal distension, increasing gas gathered in umbilical region, vanshed fat line in abdominal wall were found in 4 cases. Conclusion Large part of typical cases could be diagnosis by x-ray and clinical appearance.

Objective To evaluate MRI findings and X ray appearance in primary myelofibrosis (PMF) in marrow of lumber vertebra and pelvis. Methods Eleven cases of PMF with integrated clinical and pathology data were collected. All cases were examined with X ray and MRI simultaneously. The imaging findings of PMF were studied and compared with that of normal group. The MRI findings and X ray appearance of cases with PMF were evaluated and compared with each other to find their good quality, the shortcoming, and the relationships. Results X ray appearance in the plain film in cases with PMF: There were no abnormal findings in lumbar vertebra, iliac bone, and superior segment of femur in 5 of 11 cases. Blurred bone trabecula and ground glass changes in bone marrow were found in 3 of 11 cases. In another 3 cases, the bone trabeculas were obscured, tightly closed or mixed together, and the density of bone marrow were widespreadly high, and the difference in it couldn′t be distinguished. MRI findings in cases with PMF: MRI findings in different parts of lumbar vertebrae, iliac bone, and femur superior segments in all 11 cases were consistent. Compared with the control group, both images on T 1 and T 2 weighted spin echo displayed diffusely and well distributed lower signal intensity in 6 of 11 cases, but displayed scattered spot and patchy lowersignals in 5 of 11 cases in all parts of bone marrow including lumbar vertebra, iliac bone, and superior segment of femur. The extents of lower marrow signal intensity varied in different cases. The high signal in femoral head and greater trochanters remained unchanged as that of subcutaneous fat in 4 of 11 cases. The positive rate was 54.6%(6/11)in X ray findings, but 100%(11/11)in MRI appearances. Conclusion MRI has greater value than X ray in the early diagnosis of PMF. The early diagnostic value of X ray plain film for PMF is limited.