Objective To discuss the significance of the application of real-time quantitative PCR(RQ PCR)for diagnosis and guidance therapy of cytomegalovirus(CMV)infection after allo- hematopoietic stem cell transplantation(allo-HSCT).Methods Thirty-three patients were undergo- ing allo-HSCT.After hematopoietic reconstitution,patients'peripheral blood samples were detected for CMV DNA by RQ-PCR periodically.Anti-viral therapy was begun,attenuated and stopped ac- cording to the results of CMV DNA detection.The effects of anti-viral therapy and patients' clinical results were observed.Results CMV DNA was detected in blood samples from 13 of 33 patients. There were 21 episodes in these patients.Only 1 episode wasn't controlled hecause the patient gave up the therapy.CMV DNA copies were disappeared soon or decreased and then disappeared during anti-viral therapy in the others'.Patients which had symptoms and/or dysfunction of organs were cured too.Each course of anti-viral therapy was shorter than ordinary course.Conclusions CMV in- fection can he diagnosed as early as possible by RQ PCR.To begin,attenuate and stop anti-viral ther- apy according to the results of RQ-PCR is safe.The course is shortened and the side effects of anti vi- ral drugs were attenuated.

Objective Base on the theory of traditional Chinese medicine (TCM), obtained human diagnostic information available for joint syndrome differentiation is integrated based on subjective and objective combined syndrome differentiation, in the form of data expression of TCM four diagnosis, forms multiple information fusion of open platform for TCM diagnosis and treatment, and provides some revelation for promoting the development of depression and treatment technology. Methods Totally 30 depression patients and 30 normal people were selected according to the inclusion criteria. TCM four diagnostic and auxiliary diagnosis instrument was used to collect information of four diagnosis, and the statistical software was used for the analysis on pulse diagnosis, and information features of digitalized tongue and listening diagnosis of patients in depression group and normal group were studied. Results Compared with depression group, pulse frequency, fluency, and heart rate of normal group were a bit higher than depression group, without statistical significance (P>0.05);There was statistical significance among pulse frequency, strength, tightness, and pulse wave velocity between the normal group and depression group (P<0.05). 30 patients had 6 depression syndrome types:heart-gallbladder qi deficiency (8 cases), phlegm-heat attacking internally (6 cases), fire excess from yin deficiency (6 cases), liver depression forming fire (6 cases), intense heart fire (3 cases), and heart-spleen deficiency (3 cases). Conclusion The results of differences in pulse diagram parameters were consistent with the theory of classical TCM pulse theory. The results of differences in pulse wave velocity conform to the modern medical research conclusion. TCM four diagnostic auxiliary diagnosis and treatment technology can realize the dynamic detection of depression patients with four diagnostic information, and establish diagnostic methods for depression based on digitalized four diagnostic auxiliary diagnosis.

objective To study the reationship beween drug-resistance gene mutation and drug-resistance level in M. tuberculosis. Methods 108 M. tuberculosis clinical isolated strains from sputum specimens were analyzed by PCR-SSCP and traditional drug susceptibility tests. Results the gene mutation rate of SM, RFP, INH and EMBresistance climical isolated strains was 78.5%, 68.2%, 70.5% and 48.6% respectively, and the mutation rate of SM, RFP, INH and EMB high concentration resistance isolated strains was 86.5% , 89% , 84% and 48.6% respectively, but 28.5 % , 16.6% and 7.1% was the mutation rate of low concentration resistance strains. Conclusion The gene mutation was in relation with drug resistance level of M. tube rculosis. The gene mutation rate was hiher in high concentration resistance isolated strains than in low concentration resistane isolated strains.

Objective To observe the effects of static pressure on the number of cultured retinal M?ller glial cells(RMGC)and expression of glial fibrillary acidic protein(GFAP)and heat shock protein(HSP)70 by these cells.Design Experimental study. Participants Cultured rat RMGC.Methods Rat RMGCs were cultured and identified according to previous method described by Reichenbach.These cells were treated with different static pressures and divided into 4 groups:A(1.33kPa),B(2.67kPa),C(5.33kPa)and D(10.67 kPa)while the cells without treatment was as control group(NC).The morphologies of RMGC in these groups were observed under inverted phased contrast microscope,the number of RMGC counted with conservative method and the viability were studied with trypan blue staining.The expressions of GFAP and HSP70 in RMGCs were detected with the method of western blot.Main Outcome Measures The morphologies of RMGC,cell number,cell viability.Results There were pressure-dependent changes of RMGC number. The cell number of group C and D was less than that of group NC,A and B(P＜0.01).High static pressure resulted directly in the decreased ratio of unstained RMGCs(P＜0.01).The ratio of unstained RMGCs in group C and D was less than that in group NC,A and B(P＜0.01).Many cells in group C and D were injured and the higher the pressure elevated,the more the degree of injury became.The expressions of GFAP and HSP70 in group NC were less than other pressure treated groups and the expression of GFAP in group C and D was higher than that in group A and B.There was no obvious difference between these pressure treated groups.Conclusions High static pressure could cause the injuries of RMGCs.The increased expression of GFAP and HSP70 in RMGC might be regarded as a sign of retinal injury response to high intraocular pressure.

A reverse transcription polymerase chain reaction (RT-PCR) and single-strand conformation polymorphisms (SSCP) assay were applied to rapidly detect the molecular variability in CP and SP genes among seven isolates of Rice stripe virus in China. The PCR results showed that the CP gene of JD isolate and SP gene of PJ isolate could not be amplified. SSCP analysis showed that there were completely different electrophoretic pattern of CP gene among six isolates. To SP gene, SSCP results also discovered polymorphisms. There were five patterns among these isolates, and the pattern of YL and BS isolates were same.

Objective To develop a nylon membrane chip for rapid and systematic detection of the diabetes-associated 45 mutant loci in mitochondrial DNA(mtDNA).Methods The mutant-and wild-type probes were designed for detection of 45 mutant loci in mtDNA with Primer Premier 5.0 and NCBI BLAST softwares and the 90 probes with 8 poly T were immobilized on the Hybond N~+ nylon membranes which were treated with 5?SSC Buffer by UV-crosslinking;Then asymmetric PCR was employed to obtain the target single strand DNA(ssDNA).The PCR products were labeled with biotin after purification.NBT/BCIP was used as substrate that yields a very intense purple signal followed by AP-avidin,and the signals were observed in 24 samples with known sequences to evaluate the chips,each sample was repeatedly measured three times.Results The specific target fragments of 45 loci can be amplified under the same condition with nine sets of primers.The annealing temperatures of the wild-type [(59.01?1.42)℃] and mutant-type [(59.34?1.29)℃ ] probes are so close(t=1.046,P =0.301)that hybridization can be performed at the same temperature.The spots on the membrane chip are distinct,regular and well-distributed.The results of positive-and negative-control are perfect.The signals of negative probes and the background are similar.The results of chip were nearly concordant with that of DNA sequences(?~2=113.132,Kappa value =0.888,P = 0.000)except for T16189C mutant.Conclusions We have successfully developed a nylon membrane chip for rapid and systematic detection of the diabetes-associated 44 mutant loci in mtDNA.It could be used for screening for diabetic patients and high-risk people.

Objective To analyze negative lymph nodes of 34 non-small cell lung cancer(NCLC) patients with total correction by means of fluorescent quantitation PCR and immunohistcchemistry,and to form molecular bi- ology staging.Methods Clinical data and tissue samples of 193 lymph nodes were collected from 34 patients under- going resection for non-small cell lung cancer.Using fluorescent quantitation reverse transcription-polymerase chain reaction(RT-PCR) and immunohistochemistry method,lymph nodes were examined for CEA gene mRNA,P53 and CK to form molecular biology staging.All the patients were followed-up for an average of forty months.Results The CEAmRNA was identified in 21.7% (42/193) lymph nodes negative patients from 17 patients(17/34,50%); TMN staging was up-regulated in 8 patients;positive lymph nodes were increased in 9 patients.P53 and AE1/AE3 were identified 9.8%(19/193) from 11 patients,18.6 % (36/193)from 15 patients,separately;TMN staging was up-regulated in 2 patients of P53 examination and 5 patients of AE1/AE3 analysis;positive lymph nodes were in- creased in in 7 patients of P53 examination and 11 patients of AE1/AE3 analysis.There was obvious statistical sig- nificance in them,but the molecular biology staging based on the three markers was not an independent factor on re- currence and metasis of lung cancer.Conclusion CEAmRNA.P53 and AE1/AE3 analysis could find lung cancer micrometasis more sensitively to form molecular biology staging which was relative to the prognosis,but not an inde- pendent prognostic indicator.It might be good to the therapy strategy after operation.

To demonstrate the possibility of radiotherapy for echinococcosis of rats and to explore its mechanism of action, the effects of different doses of 6 MV X-ray radiotherapy on the activity of Echinococcus granulosus in rats were investigated. After being irradiated by 10, 20, 30 and 40Gy of 6 MV X-ray, a lot of examinations were carried out, such as examination of the ultrastructure of the Echinococcus granulosus cysts in rat with electron microscope, the total amount of proteins and Ca2+ ion in hydatid cyst fluid(HCF) .The potassium-pyroantimonate(PPA) cytochemical method was used to demonstrate whether the blocked calcium channels would be one reason for radiotherapy on Echinococcus granulosus cysts in rat. It was found that the ultrastructures of E.granulosus cysts showed different extents of alterations or damages with abnormal changes and destruction in tissues or cells of cysts. The total protein amount in HCF was increased, while Ca2+ ions in HCF were reduced obviously in the treated groups of rats , especially in high dose groups. With PPA, some electron-dense precipitates were observed on the mitochondria and endocytoplasmic reticulum in the treated groups. It is evident that the structure of cysts of Echinococcus granulosus in rat can be damaged by radiotherapy with certain extent of the quantity-efficiency relationship.

Objective To determine the length of warm ischemic (WI) tolerance in bronchial graft from non-heart-beating donors. Methods Forty-eight rats were randomly divided into 4 groups (each group having 12 rats) according to different WI durations including WI-0 min (group A), WI-30 min (group B), WI-45 min (group C) and WI-60 min (group D). In each group, the tracheae from 6 rats were respectively imbedded in greater omentum of other 6 rats, and 14 days later, the transplanted tracheae were taken from recipients to evaluate epithelial thickness and regeneration. Results Epithelial thickness and the degree of epithelial regeneration had no significant difference (P >0. 05) between the syngeneic control group and the WI-30 minutes group. All of the grafts with WI duration of 45 min were viable, but the epithelium was significantly thinner than that in the syngeneic control group (P<0. 05). However all of the grafts with WI duration of 60 min showed lower viability rate. Conclusion The time limits of tolerance to WI of tracheal grafts from NHBDs may be 45 min.

Objective To analyze the musculoskeletal injuries related to touch screen VDT operation and design implications.Methods The effects of touch screen size and angles on touch-screen-VDT-operation-related muscle load and fatigue were explored using thorough experiment and EMG acquisition method,and the independent variables included the size and angle and the dependant variables consisted of the load and fatigue of flexor digitorum superficialis (FDS),extensor digitorum communis (EDC),extensor carpi radialis (ECR) and extensor carpi ulnaris (ECU).Results No significant difference was found with regard to pointing success rate and accuracy at all screen sizes and angles levels.FDS and EDC MVC％ increased with increasing touch screen size at all levels of angles.FDS MVC％ decreased while EDC MVC％ increased with inclining angles at all levels of touch screen sizes.All measured muscles' MF did not decrease with time.Conclusion This study helps to provide basis for the optimization of equipment design,reduce exposure to musculoskeletal injuries risks and implement primary prevention.