Objective To evaluate the performances of glucose oxidase immobilized on hydrophilic-hydrophobic silica gel, which may be employed to prepare glucose sensor for the determination of glucose in body fluids. Methods The silica gel was prepared from precursors ?-aminopropyltrimethoxysilane (APTMOS) and methyltrimethoxysilane (MTMOS) by sol-gel technique. Glucose oxidase (GOD) was covalently attached to the silica gel via carbodiimide coupling reaction between a carboxylic acid group on enzyme and an amine group of the silica gel under the participation of the linking reagents 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide. The performances of immobilized GOD were explored. Results The optimum conditions were obtained as follows: volume fraction of APTMOS 70%, enzyme content given 16 800 U, the temperature of 35 ℃ and buffer pH 5.5. The decrement in the activity of immobilized GOD for the first 2 weeks was less than 10% of its original activity, and the activity of immobilized GOD retained more than 75% of its original activity after 1 month of testing. Six independently prepared immobilized GOD on the silica gel resulted in an average bioactivity of 1 290.9 ?mol?min -1?g -1 with an R.S.D. of 3.4%. The Michaelis constant (K m) of immobilized GOD was 9.1 mmol?L -1. Conclusion Immobilizing GOD on the silica gel via the formation of peptide bonds is an outstanding enzyme immobilization method.

ABSTRACT:Objective To prepare polymeric nanomicelles capable of simultaneously loading doxorubicin (DOX) and Bcl‐2 small interfering RNA (Bcl‐2 siRNA ) , and to explore their in vitro cytotoxicity and cellular uptake in MCF‐7 human breast cancer cells .Methods Copolymer poly (ethylene glycol )‐g‐polyethylenimine‐g‐poly(γ‐benzyl‐L‐glutamate) was synthesized by the combination of reductive amination and carbodiimide methods , and its chemical structure was verified by 1 H NMR .Empty and drug‐loaded copolymeric nanomicelles were prepared by dialysis method and characterized by transmission electron microscope and dynamic light scattering .The ability of the nanomicelles to compress Bcl‐2 siRNA was measured by by agarose gel electrophoresis method . The release profiles of DOX and Bcl‐2 siRNA from the nanomicelles were explored by means of fluorescence spectrometry and dialysis method .The in vitro cytotoxicity and cellular uptake of DOX and Bcl‐2 siRNA co‐loaded nanomicelles in MCF‐7 human breast cancer cells were characterized by MTT assay and confocal laser scanning microscopy , respectively .Results The critical micelle concentration of the copolymer was about 4 mg/L ,and the sizes of self‐assembled empty and drug‐loaded nanomicelles were smaller than 200 nm .The drug‐loading efficiency and drug‐loading content of DOX in the nanomicelles were 88 .7% and 15 .1% ,respectively .The DOX‐loaded nanomicelles could efficiently compress Bcl‐2 siRNA when an N/P ratio was ≥64 .The zeta potential of DOX and Bcl‐2 siRNA co‐loaded nanomicelles was +30 mV .The release behavior of the cargoes from the nanomicells was pH‐sensitive , and the release of Bcl‐2 siRNA was more sensitive to acidic pH than that of DOX . The nanomicelles could simultaneously deliver DOX and Bcl‐2 siRNA into MCF‐7 cells , and the co‐delivered DOX and Bcl‐2 siRNA significantly increased the cytotoxicity of DOX (P<0 .05) .Conclusion The polymeric nanomicelles can co‐load DOX and Bcl‐2 siRNA and deliver them into MCF‐7 cells , and DOX in combination with Bcl‐2 siRNA can synergistically inhibit the growth of MCF‐7 cells and promote cell apoptosis ,suggesting that the nanomicells may be a promising carrier for the co‐delivery for chemotherapeutics and genes .

Objective To evaluate the feasibility of diffusion kurtosis imaging (DKI)in the diagnosis and aggressiveness assessment of prostate cancer.Methods The MR data with multi-b-value diffusion-weighted imaging (DWI)of 30 male patients with prostate cancer were retrospectively analyzed.D maps and K maps were generated from the DKI model and ADC maps were obtained using the conventional monoexponential model.Differences in the D,K and ADC values between prostate cancer and benign peripheral zone (PZ)tissues,among tumors with different Gleason scores were analyzed.P<0.05 was considered statistical significance.Results ADC and D values were significantly lower in the prostate cancers than in the benign PZs (P <0.001).K values were significantly greater in the prostate cancers than in the benign PZs (P <0.001).ADC,D and K values were different significantly among the tumors with Glea-son scores of 6,7 and ≥8 (P <0.001,P =0.01 5 and P <0.001,respectively).Conclusion The DKI model can better describe the signal intensity attenuation with b values increasing,which is helpful for diagnosing and grading prostate cancer.K value may be used to quantitatively evaluate the complicated microstructure of prostate cancer.

Objective The paper is an attentative effort to evaluate the reaction and mechanism of estrogen on pregnant rabbits with acute lung injury caused by hemorrhagic shock. Methods Sixty pregnant New Zealand white rabbits were randomly divided into 6 groups, with 10 rabbits in each group, namely normal control group (NG group, with anesthesia only), estrogen group (E2G group, with additional estrogen injection at 60 min) and the other four hemorrhagic shock groups underwent hemorrhagic shock (i.e. E2SG, FSG, SBSG, E2SBSG group;mean blood pressure-40 mmHg(1 mmHg=0.133 kPa)by phlebotomy for 15 min. After maintenance of the pressure for 45 min, the rabbits were treated with E2(0.37 mg/kg), fructose injection(5%,2 ml/kg), the p38 mitogen-activated protein kinases(p38MAPK) inhibitor SB-203580 (2 mg/kg) or E2 plus SB-203580. Tumor necrosis factor alpha(TNF-α), interleukin-6(IL-6) were measured at different time points(0 min, 60 min, 80 min and 260 min), lung tissue methane dicarboxylic aldehyde(MDA) level, lung tissue myeloperoxidase(MOP), superoxide dismutase(SOD) activity, lung tissue dry weight/wet weight (DW/WW) value were measured after the experiment was finished, pulmonary pathology of the rabbits was observed. Result (1) Serum TNF-α level of NG group and E2SG group were not significantly different compared with the other four groups at the 0 min and 60 min. At 80 min and 260 min of experiment, serum TNF-αlevel of all the four shock groups were increased, E2SG group [(172.4±16.0) and (216.7±18.6) ng/L], FSG group [(171.6 ± 9.1) and (263.9 ± 7.8) ng/L], SBSG group [(172.8 ± 7.2) and (300.6 ± 4.8) ng/L], E2SBSG group [(167.9±4.8 )and (261.8±9.6) ng/L], and significantly higher than NG group and E2G group, separately (P<0.05). (2) Serum IL-6 level of NG group and E2SG group were not significantly different compared with the other four groups at the 0 min, 60 min and 80 min. At 260 min, the serum IL-6 level[(98.3 ± 0.9) and (110.4 ± 1.8) ng/L;(120.9 ± 2.3)and (109.8 ± 2.6) ng/L] of the four shock groups (E2SG, FSG, SBSG, E2SBSG group) were significantly higher than NG group and E2G group (P<0.05). (3) Lung tissue MDA level [(2.20± 0.12),(2.57±0.11),(3.17±0.08), (2.75±1.09) nmol/mg] and MPO activity [(4.45±0.25),(6.65±0.56),(9.55±0.30), (6.78 ± 0.11) U/mg] of the four shock groups (E2SG, FSG, SBSG, E2SBSG group) were higher than NG group and E2G group (P<0.05). (4) Lung tissue SOD activity [(51.8 ± 1.8),(40.2 ± 1.5), (30.0 ± 1.7),(41.2 ± 2.0) U/mg] was significantly higher in the four shock groups(E2SG, FSG, SBSG, E2SBSG group) compared with NG group and E2G group (P<0.05). (5) Lung tissue DW/WW value(0.143 ± 0.008, 0.127 ± 0.008, 0.109 ± 0.006, 0.125 ± 0.008) was significantly lower in the four shock groups(E2SG, FSG, SBSG, E2SBSG group) compared with NG group and E2G group (P<0.05). (6) Lung tissue of the rabbits in NG group and E2G group is basically normal without obvious pathology changes. Lung tissue pathological damage of rabbits was observed in the four shock groups, and the pathological damage of rabbits in SBSG group was most serious. Conclusion Estrogen can reduce acute lung injury of pregnant rabbits with hemorrhagic shock, the p38MAPK pathway plays a critical role in mediating the salutary effects of E2 on shock-induced acute lung injury.

Objective .To prove aristolochic (AA) caused vascular endothelial cells (VEC) injury via intracellular calcium overloa-ding and investigate the mechanism of calcium dobesilate antagonism. Methods Human umbilical vein endothelial cells (HUVEC) were cultured in vitro, and randomly divided into three groups: Control group, AA group, intervention group. Microscope and transmission elec-tron microscopy were used to observe changes of cell morphology and ultrastructure. ELISA method were applied to determine thrombomedu-lin (TM) in cell culture supernatant, fluorescent indicator FLuo-3/AM and intracellular calcium concentration ([Ca2+]. Results TM val-ue and average [Ca2+] i of AA group were significantly higher than that of control group (P < 0.05). Compared with the AA group, when the concentration of calcium dobesilate was 25 μM or 50 μM, TM value and average [Caz +] significantly decreased in intervention group (P < 0.05). Compared with control group, endoplasmic reticulum was pool expansion shaped, and mitochondrial cristae was absent in AA group cells. Endoplasmic reticulum and mitochondria patterns in the intervention group cells showed some improvement, compared with AA group. Conclusion AA induced VEC calcium overloading, 'I'M secretion and injury of endothelial ceils, endoplasmic reticulum and mito-chondria destruction. Dabesilate calcium could protect endoplasmic reticulum and mitochondria and reduce AA induced VEC calcium over-loading, and these could protect VEC.

ObjectiveTo synthesize 99Tcm labeled hydrazine-nicotinamide ( HYNIC)-c (RGDfK)and evaluate its biodistribution and imaging in the severe combined immunodeficiency (SCID) nude mice bearing human lung adenocarcinoma.Methods( 1 )Tcm-HYNIC-c(RGDfK) was prepared by a two-step method using tricine and ethylenediamine diacetate (EDDA) as coligands and HYNIC as the dual functional chelator.The bioactivity of 99Tc m-HYNIC-c (RGDfK) was measured by cell binding experiments.(2) The nude mice bearing human A549 lung adenocarcinoma were randomly divided into 7 groups with 5 in each group.The 7 th group was the competitive inhibition control group and was administrated 100 μg HYNIC -c (RDGfK) 30 min earlier before the injection of 99Tcm-H Y N IC-c ( RGDfK ).The nude mice were scanned at 0.5,1,2,4,8 and 12 h respectively after intravenous injection of 7.4 MBq 99Tcm-HYNIC-c(RGDfK).The biodistribution of the agent was measured as ％ ID/g.The uptake ratio of tumor to muscle (T/NT) was also measured by placing ROI on 99Tcm-HYNIC-c(RGDfK) SPECT imaging.(3)Gamma imaging was performed in 6 mice including 3 in the competitive inhibition control group at 0.5,1,2,4,8 and 12 h post injection.ResultsThe labeling yield of 99Tcm-HYNIC-c(RGDfK) was more than 90％,and the radiochemical purity was more than 95％.99Tcm-HYNIC-c(RGDfK) can specifically bind with A549 adenocarcinoma cells with a binding rate up to 36.14％.Biodistribution study showed that the uptake in the kidney was above 20 ％ ID/g during 0.5 - 8 h post injection.The ％ ID/g in tumor was 10.52 ± 1.48 at 0.5 h,17.26 ±2.81 at 8 h,and 8.93 ±0.90 at 12 h.However,the ％ ID/g in tumor was only 2.29 ±0.85 in the competitive inhibition control group at 0.5 h.The highest T/NT was 6.87 at 8 h by the ROI analysis.Xenograffted tumors could be visualized at 1 h and delineated more clearly from 4 to 8 h post injection of 99Tcm-HYNIC-c(RGDfK).Conclusions99 Tcm-HYNIC-c (RGDfK) can be readily synthesized.Its binding with A549 lung adenocarcinoma cells is specific and the binding rate is high.

OBJECTIVEBy measuring airflow and ventilation distribution of ward building, to explore and verify the hypothesis of airborne transmission and risk factor of severe acute respiratory syndrome (SARS) nosocomial infection.

METHODSTracer gas (perfume of plant oil) was emitted to the bathroom of wards when SARS index patient lived. Six different experimental situations were designed to control the status of exhaust fan in bathrooms, exhaust fan in the top of building and fresh air exchange system. The concentration of perfume was separately measured by 4 groups of lab workers and recorded blindly by the scores of "tenth degree".

RESULTSTracer gas was detected from the wards of 8th to 13th floor.

CONCLUSIONArchitecture and ventilation system of the inpatient building in the hospital contributed to the aerodynamic condition of SARS nosocomial infection through airborne transmission. The distribution of tracer gas in the wards was associated with SARS patients in this building. It was possible that SARS could have been transmitted to for distance by aerosol or other carriers.

Air Microbiology ; China ; Cross Infection ; etiology ; Hospitals ; Humans ; SARS Virus ; isolation & purification ; Severe Acute Respiratory Syndrome ; transmission ; Ventilation

To explore the clinical features, risk factors an d treatment of retinoic acid syndrome (RAS) in patients with acute promyelocytic leukemia (APL) treated with retinoic acid, the clinical and laboratory data of 11 APL patients with RAS were retrospectively analysed. The results showed that earlier and more common symptoms of RAS were successively dyspnea (11/11), fever (10/11) and hydrothorax (6/11). Higher WBC count (> or = 15.0 x 10(9)/L) in the course of treatment of all-trans retinoic acid susceptible to develop RAS (9/11). The RAS patients were treated with dexamethasone without discontinuing the treatment of retinoic acid, complete remission was achieved in 10 cases and one patient died from disseminated intravascular coagulation. It is concluded that the identification and dexamethasone treatment of RAS in earlier period are extremely important for obtaining better clinical curative effect, and it does not influence therapeutic effect of continuing application of retinoic acid.
Adolescent ; Adult ; Child ; Dyspnea ; etiology ; Female ; Fever ; etiology ; Humans ; Hydrothorax ; etiology ; Leukemia, Promyelocytic, Acute ; drug therapy ; Male ; Middle Aged ; Syndrome ; Tretinoin ; adverse effects

[Objective]To investigate the efficacy and cost analysis ofmicrosurgical clipping and vascular embolization in the treatment of patients with cerebral aneurysms.[Methods]Two hundred and fitly-one patients with cerebral ameurysms,admitted to our hospital from January 2007 to December 2010,were collected in our study;170 patients (174 aneurysms) received microsurgical surgical clipping (clipping group) and other 181 patients (185 aneurysms) were performed vascular embolization (embolization group).Duration of hospitalization,vascular occlusion rate,intraoperative rupture rate,hospital mortality and happening of complications were compared between the 2 groups;and the efficacy were evaluated by modified Rankin scale (mRS) and cost analysis of the 2 treatments were compared 3 months after the surgeries.[Results] The length of hospital stays and cerebral aneurysm rupture rate in patients of the embolization group were significantly shorter/lower than those in the clipping group (P＜ 0.05);the scores of mRS showed no significant differences between the 2 groups (P＞0.05).The cost and sensitivity analysis indicated that the ratio of average cost/good prognosis rate in the clipping group was smaller as compared with that in the embolization group.[Conclusion] The efficacies of microsurgical clipping and embolization in the treatment of cerebral aneurysms are considerable,and both of them have their advantages and disadvantages respectively;however,the microsurgical clipping is cost-effective and economical.

In order to evaluate the heavy metal potential pollution of soil and medicinal materials in main producing area of Phellodendron amurense, we collected 32 soil samples and 32 herb samples from northeast and north of China covering four provinces. In this study, the detection of heavy metal contents was conducted by ICP emission spectroscopy and atomic fluorescence spectrometry. The results showed that the soil from all areas of Ph amurense generally reached the national standard. As, Hg, Cr, Cd, Pb and Cu content of herb samples met the requirtment of the national standard except Hg content exceeding standard slight in a few samples. The reason of excessive Hg was the ability of Hg accumulation in Ph. amurense and atmospheric environment was polluted. So, national standard and Good Agricultural Practice (GAP) must be carried out severely in Ph. amurense resources production.