The development of immunotoxin DT386-GMCSF, a fusion protein which bears the N-terminal 386 amino acids of diphtheria toxin and human granulocyte-macrophage colony-stimulating factor (GM-CSF) and targets the GM-CSF receptor (GM-CSFR), has provided a promising alternative therapy to the acute myeloid leukemia (AML). However, the poor expression of the protein in E.coli is still a bottleneck which limits the industrial production. To identify the critical down-regulating factors on the expression of DT386-GMCSF, a series of truncated mutants of DT386-GMCSF at the C-terminal of GM-CSF were generated and expressed in E.coli. The results showed that the encoding sequences for the L114 of the GM-CSF dramatically impact the expression of DT386-GMCSF. On this basis, a serial of mutants integrating amino acid substitutes were generated. The results revealed that the expression level of the mutant DF123GVT, which harbors the amino acids 1-123 of GM-CSF whose L114L115V116 was substituted with G114V115T116, was evidently higher than that of the DT386-GMCSF, whereas the specific cytotoxicity to blast recovered from mice injected with HL60, a cell line highly expresses the GM-CSFR, was similar. These results have provided an important basis for the future development of the immunotoxins targeting the GM-CSFR.

AIM: The effects of BDM on isolated rat heart in cold cardioplegia were studied METHODS: Rat heart were subjected to cold cardioplegia at 4℃ for 8, 18 and 24 h Then each heart was perfused (90 cm H 2O) in Langendorff model at 37℃ for 40 min In the high K + group( n =24) the hearts were preserved in St Thomas cardioplegic solution, in BDM group( n =24) hearts were preserved in K-H solution with BDM 30 mmoL/L RESULTS: After 18 h, heart rate and the coronary flow in BDM group were significantly higher than in high K + group( P

BACKGROUND: Mild cognitive impairment (MCI) in elderly people is becoming increasingly a new hot spot for the clinical study of Alzheimer disease at present, and it is still unknown how many cases of MCI will convert into Alzheimer disease. OBJECTIVE: To analyze the incidence rate of mild cognitive impairment and the conversion rates into dementia and Alzheimer disease among elderly people.DESIGN: A prospective cohort study based on diagnosis. SETTING: Second Department of Cadre's Ward, Bethune International Peace Hospital of Chinese PLA.PARTICIPANTS: The subjects were 216 MCI patients and 2 302 subjects with normal cognition selected in the baseline investigation to the retired cadres aged 60 years and above, in 26 military cadres' sanatoriums of Shijiazhuang city between August and September 2001.METHODS: The MCI diagnosis was according to the MCI diagnostic standard in Diagnostic and Statistical Manual of Mental Disorders - Fourth Edition (DSM-Ⅳ), and the diagnosis of dementia was according to the standard in DSM-Ⅳ revised by American Psychiatric Association. Alzheimer disease was diagnosed according to the standards of National Institute of Neurological and Communicative Disorders and Stroke/the Alzheimer Disease and Related Disorders Association (NINCDS-ADRDA). All the 216 MCIpatients and 2 302 subjects with normal cognition were enrolled in the 3-year cohort study. MAIN OUTCOME MEASURES: The incidences of MCI among subjects with normal cognition; the annual conversion rates into Alzheimer disease in MCI patients and subjects with normal cognition; the relative risks (RR) and 95% confidence interval (95% CI) of MCI and normal cognition developing into Alzheimer disease. RESULTS: The MCI group and normal cognition group were followed up once every year for 3 years. Of the 216 MCI patients, 7 died, and 209 were followed up actually, and the follow-up rate was 96.8%. Of the 2 302 subjects with normal cognition, 36 died, and 2 266 were followed up actually,and the follow-up rate was 98.4%. ① In the elderly subjects with normal cognition, the annual incidence of MCI was 4.8%, and those of dementia and Alzheimer disease were 1.3% and 0.8 respectively. ② The annual in cidence rates of dementia and Alzheimer disease in MCI patients were 8.1% and 5.6% respectively, which were insignificantly different between males and females (P ＞ 0.05). ③ The incidence rates of dementia and Alzheimer disease showed a descending trend with the elevation of educational level in both the MCI patients (trend x2 =5.57, P=0.02; trend x2 =4.92, P=0.03) and the subjects with normal cognition (trend x2 =23.1, P＜ 0.001; trend x2 =18.0, P ＜ 0.001). ④ The annual incidence rates of dementia and Alzheimer disease showed an ascending trend with aging in both the MCI patients (trend x2 =14.6, P ＜ 0.01; trend x2 =13.9, P ＜ 0.01) and the subjects with normal cognition (trend x2 =32.3, P ＜ 0.01; trend x2=23.8, P ＜ 0.01). ⑤ The relative risks of the conversion into dementia and Alz heimer disease in MCI patients were 6.4 and 7.4 times as many as those in the subjects with normal cognition.CONCLUSION: The risks of conversion into dementia and Alzheimer disease in elderly MCI patients are far higher than those in elderly people with normal cognition. It should be reinforced to monitor elderly MCI patients, who are the of high risk group of dementia.

OBJECTIVETo evaluate the efficacy and safty of photoselective vaporization of prostate (PVP) in the treatment of benign prostatic hyperplasia with obstruction within 5 years.

METHODSFrom December 2004 to December 2009, there were 782 cases have been except for neurogenic bladder dysfunction and prostate cancer, who received PVP surgical treatment of BPH. The surgical conditions and postoperative follow-up data were recorded and the follow-up cut-off time for surgery after 5 years.

RESULTSA total of 782 patients with BPH who underwent PVP were included in this retrospective study. The operation in 740 cases was successfully completed at one time. But in other 42 cases, the twice operation was performed. The mean operation time was (85 ± 38) minutes, and the mean energy delivery was (355 ± 124) kJ. The mean catheterization and postoperative hospitalization time was (2.3 ± 1.7) days and (5.2 ± 2.6) days, respectively. No severe intraoperative complications were observed. The mean follow-up was (44.1 ± 19.3) months. The shortest follow-up was 6 months. The longest follow-up was 5 years. Complete follow-up data were available for 398 of the 782 patients. Of the 398 patients followed up for 5 years, the mean international prostate symptom score after 5 years was 12.8 ± 6.9, quality of life score was 2.2 ± 1.6, maximal flow rate was (14.5 ± 2.4) ml/s, and residual urine volume was 58 ml (M50). The retreatment rate because of BPH was 2.3% (9/398). Urethral stricture and bladder neck contracture were observed in 1.5% and 0.5% of the patients, respectively.

CONCLUSIONSPVP has demonstrated remarkably consistent results for objective and subjective voiding parameters. Its late complication is rare and retreatment rate is low.

Aged ; Follow-Up Studies ; Humans ; Lasers, Solid-State ; Male ; Prostatic Hyperplasia ; surgery ; Retrospective Studies ; Transurethral Resection of Prostate ; Treatment Outcome

OBJECTIVETo find the potential interference factors for the detection of NT-proBNP and BNP in patients with chronic heart failure.

METHODSEP15-A2 issued by Clinical and Laboratory Standards Institute (CLSI) was employed to compare the precision and accuracy of commercial NT-proBNP and BNP analyzer electrochemiluminescence immunoassay system Cobas E601 and chemiluminescence system ADVIA Centaur. Moreover, NT-proBNP and BNP were detected in different time interval and in different interfered sampling conditions (haematolysis, choloplania, lipemia). NT-proBNP and BNP of 203 patients with heart failure or heart failure complicated with acute cerebral infarction were analyzed to find the deviation caused by patients' endogenous factors.

RESULTSThe precision and accuracy were comparable for NT-proBNP and BNP detection using Cobas E601 and ADVIA Centaur (total-CV below 2.9% and 3.5%, the deviation from definite value below 2.38% and 3.91%). The most suitable sample type for NT-proBNP and BNP detection was serum and EDTA-anticoagulant plasma. The detection results of NT-proBNP and BNP were comparable for at least 120 min post sampling and not affected by Hb (2 g/L), DB (428 µmol/L) and chyle (2000 FIU). NT-proBNP was significantly higher in heart failure patients complicated with cerebral infarction (P = 0.003) than in heart failure patients. BNP was significantly higher in heart failure grade III patients complicated with cerebral infarction (P < 0.01).

CONCLUSIONSCobas E601 and ADVIA Centaur supplied satisfactory detection of NT-proBNP and BNP in patients with chronic heart failure with strong anti-interference capacity. The diagnostic value of NT-proBNP and BNP for chronic heart failure should be analyzed objectively in the presence of complicating diseases.

Electrochemical Techniques ; methods ; Heart Failure ; blood ; diagnosis ; Humans ; Immunoassay ; methods ; Luminescent Measurements ; methods ; Natriuretic Peptide, Brain ; blood ; Peptide Fragments ; blood ; Sensitivity and Specificity ; Specimen Handling ; methods ; standards

OBJECTIVETo evaluated the safety and efficacy of hyperthermic intraperitoneal perfusion chemotherapy(HIPC) in the prevention and treatment of pseudomyxoma peritonei (PMP) recurrence after cytoreductive surgery(CRS).

METHODSStudies published in English before 2010 on HIPC after CRS for PMP were searched in PubMed database. Each study was carefully evaluated based on pre-determined criteria. Study results were comprehensively displayed in a form. A descriptive systematic review was performed.

RESULTSA total of 11 studies were included. The median survival time of patients in these studies ranged from 25.6 months to 156 months. The ranges of 1-year, 2-year, 3-year, 5-year, and 10-year survival rates were 72%-100%, 55%-96%, 59%-96%, 52%-96%, and 55%-96%, respectively. The overall complication rate ranged from 2%-15%, and the total perioperative mortality were from 0 to 7%.

CONCLUSIONHIPC after CRS is effective and safe for patients with PMP.

Chemotherapy, Cancer, Regional Perfusion ; methods ; Humans ; Peritoneal Neoplasms ; drug therapy ; surgery ; Postoperative Care ; Pseudomyxoma Peritonei ; drug therapy ; surgery ; Treatment Outcome

Diphtheria toxin A fragment (DTA) is an essential catalytic domain of diphtheria toxin (DT)-based immunotoxin. DTA protein and its antibodies play an important role in the studies on toxicology, purification and identification of DT-based immunotoxins. In this paper, DTA was expressed and purified from E. coli. After Q-Sepharose FF chromatography and (Ni+)-Sepharose affinity chromatography, 6 x His-DTA fusion protein with 90% purity was achieved. Using the purified DTA as antigen to immunize BalB/c mice, 2 hybridoma cell lines (designated as 3B6 and 3B9, respectively) secreting monoclonal antibodies (McAbs) against DTA were established. Investigations showed that both McAbs were characterized as IgG1 with titers of 1: 10(6). The binding of the McAbs to DTA was competitively inhibited by horse sera against DT. The fact that anti-DTA McAbs could be used in western blot analysis and affinity chromatography purification of DT-based immunotoxins implied that they will be useful agents in the studies on DT-based immunotoxins.
Animals ; Antibodies, Bacterial ; genetics ; Antibodies, Monoclonal ; biosynthesis ; genetics ; Chromatography, Affinity ; Diphtheria Toxin ; immunology ; Escherichia coli ; genetics ; Female ; Immunotoxins ; isolation & purification ; Mice ; Mice, Inbred BALB C ; Peptide Fragments ; immunology ; Plasmids ; Recombinant Fusion Proteins ; biosynthesis ; isolation & purification

Objective:To observe heart rate circadian rhythm in patients with chronic kidney disease (CKD) stage 5 and to analyze the effects of parathyroidectomy (PTX) on heart rate circadian rhythm in severe secondary hyperparathyroidism (SHPT) patients.Methods:A cross-sectional observation was performed in 213 patients with CKD stage 5 and 96 controls, and the patients were divided into those with severe SHPT (PTX group, n=70) and without severe SHPT (non-PTX group, n=143). Forty-six PTX patients were followed up prospectively. The baseline data were compared among these groups. Holter electrocardiogram was performed for each participant. Non-dipping heart rate was defined as night/day heart rate ratio greater than 0.9. Multiple linear regression analysis was used to analyze the related factors of heart rate circadian rhythm in patients with CKD stage 5. Results:The 24-hour, daytime and nighttime mean heart rate in patients with CKD stage 5 were all higher than those in controls, especially in PTX group (all P<0.05). The night/day heart rate ratios of controls and CKD stage 5 patients were (0.81±0.08) and (0.91±0.08) respectively ( P<0.01). Correlation analysis showed 24-hour and daytime or nighttime mean heart rate in patients with CKD stage 5 were positively correlated with serum levels of phosphorus and ln(alkaline phosphatase), while nighttime mean heart rate and night/day heart rate ratio were positively related with serum intact parathyroid hormone level. After adjusting with postoperative follow-up period (median time: 10.9 months), 24-hour and nighttime mean heart rate, and night/day heart rate ratio in PTX patients all decreased significantly (all P<0.01). Conclusions:Heart rate is increased and circadian rhythm is abnormal in patients with CKD stage 5, which are related with mineral and bone disorder. PTX significantly decreases 24-hour and nighttime mean heart rate in severe SHPT patients, and improves the heart rate circadian rhythm.

OBJECTIVETo investigate the changes of cerebral metabolism in rabbit model of hemorrhagic shock by using proton magnetic resonance spectroscopy(PMRS).

METHODSTen New Zealand white rabbits were used for construction of the model of acute hemorrhagic anemia. 1H-MRS was performed before and at the time-peint of 30, 90, and 180 min after hemorrhagic shock. The concentrations of NAA, Cr, Cho, Lac, and NAA/Cr and Cho/Cr ratios were estimated.

RESULTSHemorrhagic shock was associated with significant reductions in red blood cell count, hemoglobin level, hematocrit, pH, and PaCO, and elevations of blood lactate and PaO. The ratios of NAA/Cr at 30 min, 90 min and 180 min after shock were (1.50±0.09), (1.37±0.09) and (1.27±0.10), respectively, which were significantly lower than those before shock (2.11±0.16) (P <0.05) (1.16±0.05) and (0.97±0.04) at 30 min and 90 min after shock, respectively, which were significantly lower than those pre-shock (1.38±0.08) (P <0.05). The ratis of Cho/Cr at 30 min and 90 min were (1.16±0.05) and (0.97±0.04), respectively, which were significantly lower than those before shock (1.38±0.08) (P <0.05).

CONCLUSIONMRS can noninvasively and dynamically detect brean metabolic changes in early hemorrhagic shock, and has positive significance for early diagnosis and prognosis assessment of hemorrhagic shock.

Animals ; Aspartic Acid ; Brain ; Choline ; Disease Models, Animal ; Magnetic Resonance Spectroscopy ; Proton Magnetic Resonance Spectroscopy ; Protons ; Rabbits ; Shock, Hemorrhagic

Objective To explore the prevalence of mild cognitive impairment (MCI) among elderly veterans. Methods 2 674 veterans ( aged 60 years and over) from 26 military sanatorium in Shijiazhuang city were studied. The Mini-Mental State Examination, Global Deterioration Scale, Activity of Daily Living, Hachinski Ischemic Scale and Hamilton Depression Scale were served as screening tools. Results The prevalence of total MCI was 8 08% in elderly people. The standardized prevalence of MCI was 6 87% in male and 10 38% in female (P