EC-SOD inclusion bodies was refolded on column using metal (Ni) affinity chromatography, based on the metal-binding property of His-tag. The effect of protein amount, urea removal speed and temperature on refolding was observed. We compared the different efficiency purified with Ni-sepharose and Heprain-sepharose affinity chromatography, and studied the stability of the refolded proteins. The results indicate that the inclusion bodies can be renatured with Ni-sepharose affinity chromatography. The increase of the protein amount and urea removal rate , the lower of the renaturation efficiency. Higher temperature was benefit to protein renaturation. Both the Ni-sepharose and Heprain-sepharose affinity column can be used to purified the refolded proteins, but purified by Heprain-sepharose affinity column the protein had higher activity. The activity of renatured protein was stable in 10 ℃～50℃,when pH10 its stability was lower significantly. In denaturating solution the stability of renatured protein was low.

Objective To evaluate the relationship between damages of visual field and retinal nerve fibre layer(RNFL)thickness in primary open angle glaucoma(POAG)with high myopia.Design Retrospective,case-controlled study.Participants POAG group with high myopia(21 eyes of 17 cases),POAG group with non-high myopia(17 eyes of 16 cases),high myopia group without POAG(25 eyes of 20 cases)and normal control group(19 eyes of 17 cases).Methods The static central visual fields were tested with Humphrey 750 automated perimetry and thickness of RNFL was measured with optical coherent tomography(OCT).Main Outcome Measures Mean deviation(MD),pattern standard deviation(PSD)and mean sensitivity at upper,lower,nasal and temporal quadrants in total deviation probability plots.Thickness of RNFL at upper,lower,nasal and temporal quadrants.Results There were more obvious general depression of sensitivity in total deviation probability plots of the early POAG with high myopia than those of POAG without high myopia,and the early visual field defects of glaucoma in pattern deviation probability plots of this group.MD of POAG with high myopia was more than those of others(P＜0.05).The differences of MD,PSD and mean sensitivity between POAG with high myopia and others were significant(P＜0.05).Mean sensitivities in each quadrant of POAG without high myopia were similar to those of high myopia(P＞0.05).The thickness of RNFL of POAG with high myopia was thinner than those of others and the thickness of RNFL of normality was thicker than that of others.The correlationship between mean sensitivity and the thickness of RNFL in each quadrant was significant(P＜0.05).Conclusion Pattern deviation probability plots is an important favor for judgement of the visual field changes in POAG with high myopia.The relationship between RNFL thickness with OCT and visual field damage may provide clinically relevant information in diagnosis of POAG with high myopia

Objective To determine the plasma concentration of cytarabine(Ara-C) in children with leukemia and obtain dynamics parameters, and investigate the relationship between the parameters and clinical effect in order to provide the basis for optimization of Ara-C application. Methods Using highperformance liquid chromatogram (HPLC) to determine the plasma concentration of Ara-C, its metabolite Ara-U and infusion rate in 37 children with acute leukemia, their therapeutic reaction, remission, treatment-related infection, side-effect and long-term treatment effect were analyzed in statistic. Results Ara-C by 1～2 g/m2 intravenous drop infusion for 2 hours, the peak plasma concentration time was 2 h and peak concentration were (14.37-84.44)μmol/L, and the median was (41.42±22.80)μmol/L. The median infusion rate was 869.57at 30 minutes after Ara-C drip completion, its average level was (253.40±81.49) μmol/L, over six-times than Ara-C peak concentration. The median continuous complete remission time in 37 children was 29.8 months (5.0～53.1 months), 3y-DFS was (90.63±5.15)%. The therapy-related infection rate was 56.8 %(21/37),including three children (8.1 %) suffered from severe infection, but there was no therapy-related death and no children were off the protocol due to poor tolerance. Conclusion As post-remission treatment, high-dose Ara-C would not cause cumulation in vivo in children with acute leukemia and side-effect were slight. Ara-C could improve the long-term continuous complete remission rate and clinical cure rate for children with leukemia. Therefore, it was worth to apply in clinical.

dialysis.

The effect of addition nucleotides on heparinase production by Flavobacterium heparinum was reseached. The result indicated that addition of nucleotides cause different effects on heparinase production depend on its addi ng mode, single kind of nucleotide would restrain, but multiplex promote. The s trongest promotion happened when the propotion of nucleotides was consistent wit h the nucleic acids of heparinase mRNA. HPLC result confined the entrance of nu cleoides to the bactera cell. And it also suggested the synthesis metabolic of nucleic acids in F.heparinum was imbalance, there always be higher lever of purin nucleoides. L-Asp was added to medium culture to enhance pyridine nucle oides thynesis in order to improve the enzyme production ,which has been affecte d by the imbalance of nucleoides.

Animals ; Astrocytes ; immunology ; metabolism ; Male ; Microglia ; immunology ; metabolism ; Neurons ; metabolism ; Oxidopamine ; metabolism ; Parkinson Disease ; immunology ; Rats ; Rats, Sprague-Dawley

Objective To investigate clinical features of lower motor neuron lesion (LMNL) caused by the single level lower thoracic disc protrusion (LTDP),and to observe clinical outcomes of surgical treatment.Methods Between January 1997 and December 2009,17 patients with LMNL caused by single level LTDP underwent en bloc resection of the superior articular process,Cave-in 360° circumferential decompression and internal fixation in our hospital.MRI and CT scans were taken to confirm lesion levels:T10-11 in 4 patients of whom 3 had patellar clonus and ankle clonus,T11-12 in 5 patients of whom 4 had ankle clonus,and T12L1 in 8 patients who only had positive Babinski sign.The neurologic status was assessed using the Japanese Orthopaedic Association (JOA) scoring system.The muscle strength of the tibialis anterior was assessed using the Manual Muscle Test (MMT).Sagittal Cobb angle and cross-sectional area of the dural sac at the level of maximal compression in MRI were also observed.Results All patients were followed up for 22 to 76 months (average,48.6 months).The mean JOA score increased from preoperative 5.88±1.11 to 9.53±0.94 at final follow-up (t=16.143,P＜0.05).The muscle strength of the tibialis anterior recovered to more than grade 4 in all patients.Postoperative Cobb angle was unchanged compared with that before operation.MRI indicated that the cross-sectional area of the dural sac at the level of maximum compression increased from preoperative 35.8±7.3 mm2 to postoperative 132.9±6.5 mm2 (t=70.78,P＜0.05).Conclusion LMNL can be caused by LTDP.The eu bloc resection of the superior articular process,Cave-in 360° circumferential decompression and internal fixation can provide a satisfactory decompression effect and marked recovery of neurological function.

Objective:To investigate the change of biological characteristics after stable knockdown of CKLF-like MARVEL transmembrane domain containing 3 (CMTM3)expression in PC3 by lentivirus shRNA and to reveal new therapeutic targets.Methods:The research includes two groups:sh393 is the experimental group in which CMTM3 is knocked down in PC3 cell line;shN is the control group in which CMTM3 is negatively knocked down.The expression of CMTM3 was detected by Western blot.The mi-gration ability of PC3 after stable knockdown was detected by Transwell and Wound healing assay.The invasion ability of PC3 was detected by Matrigel assay.Results were obtained from at least three indivi-dual experiments.Results:The expression of CMTM3 in sh393 group is significant lower than shN group (0.004 0 ±0.000 4 vs.0.490 0 ±0.055 7,P <0.001)detected by Western blot.It also had statistical significance in Matrigel assays (248.6 ±4.5 vs.113.0 ±3.3),Transwell (203.6 ±1.9 vs.103.0 ± 1.2)and Wound healing assays (95.0 ±2.9 vs.33.0 ±1.5)that knockdown of CMTM3 promoted mi-gration,and invasion of PC3 cells in vitro (P <0.001).Conclusion:Negative correlation exists between the stable knockdown of CMTM3 and change of biological characteristics in PC3 cells,and knocking down CMTM3 affects migration,and invasion ability in PC3 cells.

Objective · To investigate the clinical characteristics and multidisciplinary treatment of children with clear cell sarcoma of kidney (CCSK). Methods · Data of seven children with CCSK treated at Xinhua Hospital, Shanghai Jiao Tong University School of Medicine between Jan 2011 and Jan 2016 were collected. The retrospective analysis of clinical manifestations, features of imaging and pathology, treatment, and follow-up was performed. Results · Of 7 children with CCSK, 6 were male and 1 was female with the median age of 28 months (4-59 months), and 3 were at stageⅠ, 2 at stage Ⅲ, and 2 at stage Ⅳ. All cases were discussed and evaluated by multidisciplinary teams, including pediatric hematology/oncology, pediatric surgery, pathology, radiology and radiotherapy. CCCG-WT-2009 protocol was adopted to treat these patients. The median follow-up period was 22 months (8-56 months). Six children survived and one died. Conclusion · The multidisciplinary treatment mode can effectively improve the prognosis of CCSK. CCCG-WT-2009 protocol has good therapeutic effect and high cure rate for children with early stage CCSK, but the treatment of advanced stage CCSK needs to be further explored and perfected.

Objective · To investigate the changes of neutrophils and lymphocytes during chemotherapy courses in children with high-risk neuroblastoma, as well as severe infection rate and its relationship with combination chemotherapy regimens in order to provide guidance to prevent infection in these children. Methods · A total of 45 cases of children with high-risk neuroblastoma, who were hospitalized in Xinhua Hospital from July 2012 to September 2015 were recruited. Reactions after chemotherapy and the results of routine blood tests on every 5 days during the whole chemotherapy circles were recorded. Follow-up visits were carried out to learn the infection occurrence. Results · The counts of neutrophils and lymphocytes dropped to the minimum at the 10th day or so during chemotherapy, then rised near the 15th day. Infections were negatively correlated with the numbers of neutrophils (r =-0.245) and lymphocytes (r=-0.227) at the 10th day. Persistent lymphopenia occured after the forth course of chemotherapy. Rate of grade Ⅳ bone marrow suppression after chemotherapy was 90%, and severe infection rate was 15.5%. There were significant differences in the rates of grade Ⅲ bone marrow suppression (P=0.008), grade Ⅳ bone marrow suppression (P=0.001) and severe infection (P=0.010) among the 3 combination chemotherapy regimens used in high-risk neuroblastoma children. Conclusion · Children with high-risk neuroblastoma had obviously high rates of bone marrow suppression and severe infection after chemotherapy, especially after cyclophosphamide+doxorubicin+vincristine regimens. More attention should be paid to this group of children and preventive measures were worthy of taking into account.