OBJECTIVE: To observe the expression pattern of caspase-3 and HCLS1-associated protein X-1 (HAX-1) at different time after cerebral contusion in rat, and explore the new method for estimating the injury interval. METHODS: The cerebral contusion model was established using adult SD male rats. Then the rats were randomly allocated into 8 groups: 2 h, 6 h, 12 h, 1 d, 3 d, and 7 d after cerebral contusion, sham-operation and normal control. Expression of caspase-3 and HAX-1 protein after cerebral contusion in rat was detected by Western blotting. Laser scanning confocal microscope was used to observe the number of HAX-1 positive cells and TUNEL-stained cells after cerebral contusion. RESULTS: The expression of caspase-3 increased parallelly with the time after cerebral contusion and reached the peak value on 3 d. The expression of caspase-3 decreased gradually and still maintained a high level expression on 7 d (P < 0.05). The expression of HAX-1 positive cell went up after injury, and reached the peak value at 6 h (P < 0.05), then turned down gradually after 12 h and went out of detection after 3 d. The number of TUNEL-stained cells increased obviously at 2 h and reached the peak value on 3 d. The number of TUNEL-stained apoptotic cells decreased gradually and still maintained a high level expression on 7 d (P < 0.05). CONCLUSION The expression of caspase-3 and HAX-1 after cerebral contusion has time sequential regularity, which may provide new evidence for forensic diagnosis of cerebral contusion interval.
Animals ; Blotting, Western ; Brain Injuries/pathology* ; Carrier Proteins/metabolism* ; Caspase 3/metabolism* ; Cerebellum/pathology* ; In Situ Nick-End Labeling ; Intracellular Signaling Peptides and Proteins ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo discuss the clinical effects and superiority of applying external fixation and bone transport to treat osteomyelitis and bone defect of femoral bone.

METHODSFrom August 2008 to December 2013,16 patients with osteomyelitis and bone defect of femoral bone were treated including 11 males and 5 females with an average age of 42 years old ranging from 13 to 62 years old. The average course of disease was 18 months ranging from 2 months to 4.5 years, and the average length of bone defect was 7.8 cm ranging from 4.5 to 15 cm. The bone defect of all cases were treated by external fixation and bone transport, the bone transport began at 1 week after operation, 1 mm per day and 4 times per day.

RESULTSAll patients were followed up for 10 to 36 months (means 22.5 months). One patient did not cooperate with treatment leads to the failure, then took the amputation. The remaining 15 cases of osteomyelitis were under control, including 12 cases of bone transport achieved one stage bone union, 3 cases achieved bone union via bone graft from iliac bone. The bone union time was 5 to 13 months(means 7.9 months). Thirteen patients almost obtained the same length of two lower extremities,2 patients had shortening of 1.5 to 2 cm. The time of moving the external fixation was from 6 to 16 months (means 9.3 months).

CONCLUSIONApplication of external fixation and bone transport is an effective method in treating the osteomyelitis and bone defect that can control the infection, eradicate wounds, and be the equalization of limb length.

Adolescent ; Adult ; Bone Transplantation ; External Fixators ; Female ; Femur ; surgery ; Humans ; Male ; Middle Aged ; Osteomyelitis ; surgery

Objective To evaluate the olfactory functions in patients with Parkinson's disease (PD) with a smell identification test developed for the Chinese population and to examine possible factors contributing to PD patients' olfactory performance.Methods Fifty-five patients with idiopathic and nondementia PD and 55 demographically matched healthy controls (HC) were recruited and their olfactory identification tested using the Chinese Smell Identification Test developed by the Institute of Psychology at Chinese Academy of Sciences.Results Olfactory identification score was significantly lower (t =-6.827,P＜0.01) in the PD group (19.3 ±5.4) as compared with the HC group (26.5 ±5.7).Of all PD patients,63.6％ scored over 1 SD below the age norm.In particular,of those with Hoehn-Yahr ratings between 2 to 3,77.8％ scored over 1 SD below the age norm.Receiver operating characteristic (ROC) curve analysis indicated that a cut-off score of 22.5 on the Chinese Smell Identification Test distinguished between PD patients and HC with a sensitivity of 74.6％ and specificity of 70.9％.Within the PD group,age,education,Hoehn-Yahr rating and Mini-Mental State Examination score were not found to correlate with olfactory performance (all P ＞ 0.05).Conclusion PD patients in China show significant impairment of olfactory identification,which is effectively captured by the Chinese Smell Identification Test.

Objective To investigate the effect of ropiyacaine combined with intra-spinal epidural anesthesia by using the same volume but different injection speed on anaesthetic level.Methods 80 cases of patients of ASA～Ⅱdegree suitable to use intra-spinal epidural combined anesthesia for gynecologic operation were selected and ran- domly divided into three groups,0.75 % ropivacaine 2mg(15mg)administered,group A the speed of injection was 10 seconds,group B injection speed was 20 seconds,group C injection speed was 30 seconds,the anaesthetic level reached T_(10).The time of highest level in spinal anesthesia,30 minutes after spinal anesthesia MAP,and number of cases need to add epidural drug were all recorded.Results The best effect of anesthesia was found in group B,the block level of anesthesia was satisfactory,blood dynamic was stable,and there was no need to add epidural drug.Conclusion The speed of 0.75 % ropivacaine 2ml spinal epidural combined with anesthesia was suitable at the speed of 20 seconds.

Objective To investigated the effects of combined arsenic trioxide(ATO) and resveratrol(Res)on the viability of NB4 human leukemia cells. Methods NB4 human leukemia cell was used in this experiment.Cells were cultured in ATO (0,0.1875,0.3750,0.7500, 1.1250, 1.5000,2.2500,3.0000,5.0000 μmol/L) and Res (0, 1.5625,3.1250,6.2500, 12.5000, 18.7500,25.0000,37.5000,50.0000 μmol/L). Cell viabilities were measured by MTT in different treatment groups. Half inhibitory concentration(IC50) was calculated. The ratio of concentration of ATO and Res 1.5∶ 18,1.5∶ 25,1.5∶ 35 was added to cells, and the combination index(CI) was calculated. The level of ROS in control, ATO( 1.5000 μmol/L), Res(25.0000 μmol/L) and ATO(0.9000 μmol/L) + Res( 12.5000μmol/L) groups was measured by chemiluminescence assay. Results ①ATO( ≥0.7500 μmol/L) reduced the viability of NB4 cells in a concentration-dependent manner(P ＜ 0.05 ), and IC50 was (1.78 ± 0.11 )μmol/L. ②)Res (≥18.7500 μ mol/L) dose-dependently decreased the viability of NB4 cells (P ＜ 0.05 ), and IC50 was ( 18.71 ±0.18)μ mol/L. ③Combination of ATO and Res showed an antagonistic effect on NB4 cells viability. ④The ROS in Res group( 1670.55 ± 13.97) was significantly lower than that in control group(2345.88 ± 14.48,P ＜ 0.05). The ROS in ATO group (3092.42 ± 94.84) was significantly higher than that in control group(P ＜ 0.05). The ROS in ATO + Res group (1860.27 ± 15.99) was significantly lower than that in ATO group(P ＜ 0.05). Conclusions NB4 cell survival rate can be decreased by ATO and Res. The combination of arsenic trioxide and Res presents an antagonistic effect on NB4 cell viability, in part by reducing intracellular ROS formation.

Objective To investigate the clinical outcomes of minimally invasive percutaneous nephrolithotomy(MPCNL)in the treatment of lower caliceal calculi.Methods We retrospectively re- viewed the clinical outcomes and complications of 33 patients who underwent MPCNL for lower caliceal cal- culi from March 2001 to April 2005.The average diameter of the calculi was 2.8 cm.Single tract nephrosto- my was performed in all 33 cases;among them renal access was obtained through a middle calyx in 10 cases and a lower calyx in 23.Nine cases had F14 renal access;and 24 cases,F16.Results Of 33 cases,28 (85%)achieved stone-free at 1 session.A second-look was needed in 3 cases due to intraoperative bleed- ing;ESWL,in 1 case with residual,calculi;no treatment,in 1 case with residual calculi＜4 ram.The mean operative time was 93 min;mean blood loss was 113 ml;mean hospital stay was 11 d.Blood transfusion was needed in 1 patient who suffered from hepatic cirrhosis preoperatively;another experienced severe bleeding 7 d after operation and was cured with hyperselective spongia gelatinosa embolization of the renal artery.Fol- low-up was available in 19 cases for 2-48 months,and no recurrence of renal calculi was noted.Conclu- sions Minimally invasive percutaneous nepbrolithotomy has advantages of safety,less invasion,and easy re- covery for the treatment of lower caliceal calculi.

Objective To predict monthly incidents of extended spectrum β-Lactamases (ESBLs)-producing Escherichia coli in Zhejiang Hospital by establishing multiple seasonal autoregressive integrated moving average (ARIMA) model, so as to provide scientific evidence for reducing the incidents of nosocomial infection of ESBLs producing Escherichia coli. Methods Multiple seasonal ARIMA model was established by monthly records of ESBLs producing Escherichia coli from 2010 to 2016 in Zhejiang hospital. Monthly incidents of ESBLs producing Escherichia coli from 2017 to February 2018 were used to verify the predicted result. The predictions were evaluated by models of mean absolute percent error (MAPE) and bayesian information criterion (BIC) . Results The optional model for the monthly incidence from 2010 to 2016 was ARIMA (0, 1, 1) (0, 1, 1)12. The MAPE was 14.76, BIC was 2.01, and the Ljung-Box statistics value Q was 16.79 (P=0.40) . These parameters suggested a good model fitting. The average relative error between the predictive value and the actual value of the monthly incidents ESBLs producing Escherichia coli from 2017 to February 2018 was 14.08%.The actual values were within the 95% confidence interval. Conclusion The multiple seasonal ARIMA model of ARIMA (0, 1, 1) (0, 1, 1 )12 fits and can be used for short-term prediction and dynamic analysis of the incidents of ESBLs producing Escherichia coli in Zhejiang Hospital.

OBJECTIVEAn RP-HPLC procedure was established for the determination of danshensu and protocatechuic aldehyde in Guanxinning injection powder.

METHODAn RP-HPLC analytical procedure was developed using Hypersil ODS2 C18 column (4.6 mm x 250 mm, 5 microm) with methanol-0.5% glaceal acetic acid (4.5:95.5) as mobile phase, and a wavelength of 280 nm for UV detection.

RESULTThe linear range was 3.004-45.06 microg x m(L-1) (r = 0.999 5, n = 6) for danshensu, and 0.300-4.509 microg x mL(-1) (r = 0.999 3, n = 6) for protocatechuic aldehyde. The average recoveries were 99.1% and 97.9%, respectively.

CONCLUSIONThe method was stable, accurate, and reproducible, and can be used for the quality control of Guanxinning injection powder.

Benzaldehydes ; analysis ; Catechols ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Injections ; Lactates ; analysis ; Ligusticum ; chemistry ; Plants, Medicinal ; chemistry ; Powders ; Quality Control ; Reproducibility of Results ; Salvia miltiorrhiza ; chemistry

The biological activity of ADCC by anti-CD20 monoclonal antibody was determined by BioGlo™ Luciferase Assay System using Jurkat/NFAT-luc+FcγRIIIa cell line as effector cell and WIL2-S cell line as target cell. The developed method was verified for specificity, precision and accuracy. Anti-CD20 monoclonal antibody showed a dose-response mode by the developed method, and the determination result complied with the following four-parameter equation: y = (A-D)/[1 + (X/C)(B)] + D. The optimized parameters of the method were determined including the antibodies diluted concentration (18,000 ng·mL(-1)), dilution rate (1:5), the ratio of effector cell and target cell (6:1), and induction time (6 h). The values of eight independent tests have passed a statistical test for curve regression analysis, linear or parallelism, which showed the method possessed good specificity. Four different dilute groups of recovery rates sample were determined for 3 times, and the result showed mean relative potencies of (44.39±3.93)%, (72.74±2.78)%, (128.28±7.01)% and (168.19±2.70)% respectively, with a variation coefficient of less than 10%, and the recoveries of (88.78±7.85)%, (96.99±3.70)%, (102.63±5.61)% and (112.12±1.80)% respectively. A novel reporter gene method for determination of biological activity of ADCC by anti-CD20 monoclonal antibody was successfully developed, which showed strong specificity, good reproducibility and high accuracy, and might be used routinely.
Antibodies, Monoclonal, Murine-Derived ; pharmacology ; Antibody-Dependent Cell Cytotoxicity ; Antigens, CD20 ; immunology ; Genes, Reporter ; Humans ; Reproducibility of Results ; Rituximab

OBJECTSTo investigate the distribution of vitamin D receptor (VDR) genotypes among the Hans of a lead contaminated mine in Shanxi and explore the relationship between blood lead levels and the genetic polymorphism of VDR gene.

METHODSVDR genotypes were determined by polymerase-chain-reaction and restrictive fragment length polymorphism (PCR-RFLP) and the blood lead level was measured by using the graphite furnace atomic absorption spectrometry in a population of 120 pre-school children aged 5 - 6 years who were from the mine kindergarten and were unrelated Hans. An environmental questionnaire in relation to blood lead level was filled for each subject.

RESULTS(1) The gene distribution of the VDR phenotypes in these children was VDRBB, 1.7%; VDRBb, 9.2%; VDRbb, 89.2%. (2) The mean blood lead level of the children who had VDR B allele [(0.910 8 +/- 0.265 0) micromol/L] was significantly higher than that whose VDR genotype was bb [(0.740 1 +/- 0.270 1) micromol/L (mean +/- standard deviation)] (t = 2.155, P < 0.05). (3) Many factors were found to affect the blood lead levels, such as the VDR genotype, the type of fuel, educational level of mothers and so on. After controlling the possible confounding variables by multiple regression, the contribution of the VDR phenotype to the blood lead levels was still statistically significant.

CONCLUSIONThese results indicated that the frequency distribution of the VDR genotype in these children was apparently different from that in Caucasians who had high frequencies of VDR B. The results also indicated that the individuals carrying the VDR B allele were more susceptible to lead poisoning.

Child ; Child, Preschool ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Lead ; blood ; Lead Poisoning ; genetics ; Multivariate Analysis ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Receptors, Calcitriol ; genetics ; Regression Analysis ; Surveys and Questionnaires