The method of multiplex PCR was set up to identify two or three transgenes in one reaction such as uidA and bar; uidA and IDx5 or uidA, bar and 1Dx5 genes. Three sets of primer pairs which was specific to each of these three genes respectively were designed and synthesized. Recombinant plasmids pAHC25 and p1Dx5 harboring uidA + bar and 1Dx5 gene separately were used as template DNA in the process of optimizing an multiplex PCR reaction. The optimal annealing temperature for uidA and bar MPCR is range from 57. 1℃-62. 3℃ , for uidA and 1Dx5 is range from 60℃ to 60. 6℃ , and for uidA、bar and 1Dx5 range from 57. 0℃-58. 4℃. The amount of template for MPCR is twice as much as that for simplex PCR, while the concentration of primers is the same with simplex one. Less than 50bp MPCR products can be separated clearly by 10% non-denaturalized polyacrylamid gel electrophoresis. Fourteen transgenic wheat lines were tested by multiplex and simplex PCR respectively, which shows the same results and hence presents that MPCR is the reliable, rapid and high-effective approach to detect foreign genes from transgenic plant.

Objective To investigate the effects of low frequency repetitive transcranial magnetic stimulation (rTMS) applied to the supplementary motor area (SMA) of children with Tourette's syndrome (TS). Methods Thirty TS subjects less than 16 years old were treated with 1 Hz rTMS to the SMA at 110％ of the resting motor threshold (RMT) in 20 daily sessions,receiving 1200 pulses/day.Clinical assessment and physiological measures of the left and right RMT were conducted at different time points during the treatment. ResultsAfter 4 weeks of treatment,statistically significant reductions were observed in assessments with the Yale global tic severity scale (YGTSS) and in terms of clinical global impression (CGI).Symptomatic improvement was correlated with dramatic increases in both right and left RMTs. ConclusionApplication of 1 Hz rTMS to the SMA can improve the clinical symptoms of TS children.

Objective To evaluate the clinical significance of carcinoembryonic antigen (CEA),cancer antigen 125 (CA125),carbohydrate antigen 19-9 (CA19-9) and eytokeratin 21-1 fragment (CYFRA21-1) assays of sputum in patients with lung cancer. Method Fifty-two cases with lung cancer and 46 cases with benign lung diseases underwent detection of CEA ,CA125 ,CA19-9 and CYFRA21-1 in sputum by using the method of chemi-luminescent enzyme immunoassay. Results The levels of CEA,CA125,CA19-9 and CYFRA21-1 in sputum of 52 cases with lung cancer were (27.6±31.2) μg/L, (76.4±65.2)kU/L, (56.1±31.6) kU/L and ( 25.2±9.1 )μg/L respectively. But the levels of those of 46 cases with benign lung diseases were (6.1±7.5)μg/L, (23.7±7.9) kU/L, (17.3±10.2) kU/L and (1.2±1.7)μg/Lrespectively. The levels of these tumor markers in sputum in patients with lung cancer were significantly higher than those in patients with benign lung diseases(P< 0.01 ). The diagnostic sensitivity of CEA,CA125,CA 19-9 and CYFRA21-1 in sputum in 52 cases with lung cancer was 42.3%( 22/52 ), 46.2% (24/52), 36.5%( 19/52 ) and 51.9% (27/52) respectively ( P > 0.05 ). Among the cancer patients, the sensitivity of sputum CEA in patients with adenocareinoma was significantly higher than that in patients with squamons cell carcinoma (X2= 4.193, P < 0.05 ) ; while the sensitivity of sputum CYFRA21-1 in patients with squamous cell carcinoma was significantly higher than that in patients with adenocarcinoma ( X2 = 4.806,P < 0.05 ). The sensitivity of CA125 in advanced lung cancer( Ⅲ +Ⅳ stage) was higher than that in early lung cancer( Ⅰ+Ⅱstage) (X2= 5.202,P < 0.05 ). Compared with the single tumor marker assaying, the combination of CEA,CA 125, CA 19-9 and CYFRA21-1 in sputum could significantly improve the sensitivity and accurate value in the diagnosis of lung cancer (P<0.05). Conclusion To assay CEA,CA125,CA1g-9 and CYFRA21-1 in sputum is valuable in diagnosis of lung cancer, and the combination of CEA,CA125,CA19-9 and CYFRA21-1 in sputum can significantly improve sensitivity and accurate value in the diagnosis of lung cancer.

OBJECTIVE:To establish a method for the residues determination of Pb,Cd,Cu,As and Hg in Codonopsis pilo-sula,and evaluate the quality evaluation of C. pilosula of Pingshun County in Shanxi province. METHODS:Microwave diges-tion-inductively coupled plasma mass spectrometry was adopted with KED scanning model,RF power was 1 550 W,sampling depth was 5.0 mm,plasma gas(argon)flow rate was 16.0 L/min,helium partial pressure was 0.1 mbar,argon gas was 0.6 mbar, the vacuum degree of 5×10-8 mbar,branch turbopump speed was 1 000 hz,sampling cone aperture was 1.0 mm,skimmer aperture was 0.5 mm,the spray chamber temperature was 2.7 ℃,the data collection was repeated 3 times. RESULTS:The linear range was 0-20 ng/ml for Pb(r=0.999 3),0-10 ng/ml for Cd(r=0.998 5),0-250 ng/ml for Cu(r=0.998 8),0-20 ng/ml for As(r=0.999 0) and 0-1.0 ng/ml for Hg(r=0.997 9);RSDs of precision,stability and reproducibility tests were lower than 3.0%;recoveries were 95.80%-100.20%(RSD=1.85%,n=6),94.50%-98.00%(RSD=1.26%,n=6),98.52%-102.43%(RSD=1.60%,n=6), 94.90%-98.70%(RSD=2.29%,n=6)and 96.00%-101.00%(RSD=1.84%,n=6);the limits of detection were 0.021 0,0.003 4, 0.043 7,0.115 6 and 0.005 6 ng/kg,respectively. Pb,Cd,Cu,and As were detetcted,and Hg was not detected,the range of total contents was 7.185 2~12.558 0 mg/kg. CONCLUSIONS:The method is simple with good precision,stability and reproducibility, and can be used for the residues determination of Pb,Cd,Cu,As and Hg in C. pilosula;heavy metal residues in C. pilosula in Shanxi Pingshun county does not exceed limit values of national and industry standards.

Objective To approach the clinical significance of Clara cell secretary protein(CCSP) in bronchial asthmatic children. Methods Serum were collected from 50 cases during asthmatic attacks, 22 asthmatic children who were in stable conditions, and 20 healthy children. Serum CCSP concentrations were measured by a human CCSP enzyme- linked immunosorbent assay (ELISA). Results Asthmatic children had significantly lower levels of CCSP in serum during asthmatic attacks(P

ObjectiveTo investigate the effect of focal ischemic preconditioning (IPC) on the expression of protein kinase-like endoplasmic reticulum kinase ( PERK ) and glucose-regulated protein 78 (GRP78) mRNA and protein after focal cerebral ischemia/reperfusion (I/R) in rats.MethodsAll 120 male SD rats were randomly divided into three groups: sham-operation group,middle cerebral artery occlusion (MCAO) group and brain ischemia preconditioning (BIP) group.Each group was further divided into 4 subgroups according to 12 h,1,2 and 3 d after I/R.The IPC models were made in order to measure the expression of PERK,GRP78 mRNA and protein by in situ hybridization and Western blot,and the apoptosis rate of neuron by flow cytometry. Results ①The expression of PERK mRNA increased and reached the peak at 12 h,then decreased continuously after 1 d.BIP could decrease its expression.The expression of PERK protein increased at 12 h and reached the peak at 24 h,then decreased continuously after 2 d.BIP could decrease its expression.②The expression both of GRP78 mRNA and its protein all increased and reached the peak at 12 h,then decreased continuously.BIP could increase their expression (mRNA:12 h: 136.70±9.53,F=32.265; 24 h:147.54 ±9.97,F=54.920; 2 d:158.16 ±9.44,F=45.374; 3d: 165.85±10.26,F=16.493,P＜0.05; protein:12 h: 1.319±0.116,F=5.619,P＜0.05; 24 h: 1.226±0.108,F=33.742,P＜0.01; 2 d:1.183 ±0.112,F =46.556,P ＜0.01; 3 d:1.115± 0.098,F =11.730,P＜0.05).③The rate of apoptosis neuron of rats in MCAO increased markedly at 12 h after reperfusion,and reached the peak at 1 d,then decreased continuously.BIP could decrease the rate of apoptosis neuron. Conclusion BIP can protect neurons through inhibiting the expression of PERK and inducing the expression of GRP78 after endoplasmic reticulum stress in rats.

OBJECTIVETo establish fingerprint database of drug-resistant strain of Acinetobacter baumannii with AFLP genotype.

METHODSTwenty-seven strains of A. baumannii were clinically isolated. The microbial sensitivity to 13 antibiotics was analyzed according to NCCLS 2004. The genotype was analyzed by amplified fragment length polymorphism (AFLP) method.

RESULTSThe results were consistent with those with AFLP database.

CONCLUSIONThe database of AFLP fingerprint may have practical value for identification of drug-resistant Acinetobacter baumannii.

Acinetobacter baumannii ; genetics ; Amplified Fragment Length Polymorphism Analysis ; DNA Fingerprinting ; DNA, Bacterial ; genetics ; Databases, Nucleic Acid ; Drug Resistance, Bacterial ; genetics ; Genotype ; Microbial Sensitivity Tests ; Polymorphism, Genetic

Objective To discuss the significance of the retrograde urethrography in diagnosis and treatment of urethraltrauma. Methods 78 cases with urethraltrauma treated by the retrograde urethrography were retrospectively analyzed. Results The location and extent of urethral injury was determined according to the place and speed of contrast medium overflow and the diffuse range. Among 78 cases ,29 cases were bulbar urethral trauma and other 49 cases were membranous urethral trauma.Conclusion Retrograde urethrography is simple, practical and easy to operate for determining the injured part of urethra and the extent of damage of urethraltrauma, and was instructional for the choice of operation method and incision.

Objective To study the effectiveness of personalized rehabilitation treatments based on threedimensional gait analysis (3DGA) for improving the walking function of children with cerebral palsy (CP).MethodsA total of 21 spastic CP children with diplegia or hemiplegia,IQ scores ＞60,and an average age of 8.5 years received 3DGA.They then received personalized rehabilitation treatment designed according to the 3DGA results.After four weeks of treatment the children accepted 3DGA again.Their gait descriptors before and after treatment were compared. ResultsAfter the personalized rehabilitation the subjects'clinical foot dorsiflexion angle,clinical popliteal fossa angle,walking velocity,stride length,step length,peak ankle dorsiflexion in prime stance,peaking ankle plantar flexion in last stance,peak back ground reaction forces (GRFs) and peak vertical GRF in stance all had improved significantly.The cadence,total support time,swing phase,initial double support time,peak knee extension in stance and the peak forward GRF were not,however,significantly different compared with before the personalized rehabilitation treatment.Conclusion Under the guidance of 3DGA,the walking function of spastic CP children improved significantly after 4 weeks of personalized rehabilitation treatment.3DGA can play an important role in formulating personalized rehabilitation protocols and guiding rehabilitation treatment for CP children.

Objective To study the effects of benzyl propionate nandrolone (BPN ) on the nicotinamide phosphoribosyl transferase (Nampt), insulin receptor substeate-2 (IRS-2 )and pancreatic duodenal homeobox-1 (PDX-1)expressions, cell cycle changes as well as insulin secretion in pancreatic islet cell NIT-1 lines, and to explore the influence of BPN in the Nampte xpression in NIT-1 cells and insulin signaling molecules in high glucose oxidation stress.Methods The NIT-1 cells were cultured with different concentrations (5.6,11.1,16.7,and 27.6 mmol·L-1)of glucose,then they were treated with 10 mg·L-1 BPN for 48 h with no BPN treatment as corresponding control groups.The expression levels of Nampt,IRS-2,and PDX-1 were tested by Western blotting assay.The changes of cell cycle were determined by FCM and the cell insulin secretion levels were measured with radioimmunoassay.Results Compared with corresponding control groups,the expression levels of Nampt,IRS-2, and PDX-1 proteins in the NIT-1 cells in various BPM groups were increased (P<0.05 or P<0.01).The G0/G1 phase arrest was relieved (P<0.01)when the cells was cultured in low glucose (5.6 mmol·L-1 )condition,and the G2/M block was remitted significantly in high glucose (27.6 mmol·L-1 )condition (P<0.01),furthermore, the cell insulin secretion was promoted compared with control groups except 1 1.1 mmol· L-1 glucose group (P<0.01).Conclusion BPN can promote the expression levels of Nampt,ISR-2 and PDX-1 proteins in NIT-1 cells. There is close relationship between the Nampt expression in NIT-1 cells and insulin signaling pathway and BPN prevents the cells from insulin resistance.