ObjectiveTo explore the accuracy of judging the T-staging of rectal cancer by biplane transrectal ultrasound.MethodsPathological enteroscopic biopsy was carried out for 197 cases of rectal cancer.Cases of middle and lower rectal cancer without serious obstruction were examined by means of transrectal biplane ultrasound through which to observe the changes of the echoes and to detect the depth of the infiltration in the intestinal walls and the presence of invasions in prostate glands,seminal in male or vesicles,uterine cervix in female.On the basis of the findings,the preoperative tumor staging was made.Postoperatively,the ultrasound results were compared with the pathological examination.Their consistency was evaluated by using Kappa test.ResultsTransrectal biplane ultrasound examination showed the total accuracy rate in T-staging of rectal cancer was as high as 83.8％ with the diagnostic sensitivity rate for T1to T4 as 77.8 ％,73.1％,89.3％ and 94.4％,while its diagnostic specificity rate degree was 97.3％,93.1 ％,85.1 ％,97.8％ respectively,The k -value was 0.732 which suggested that the ultrasound staging was highly consistent with the pathological staging(P＜0.05).ConclusionsTransrectal biplane ultrasound has a great value in the preoperative T-staging of rectal cancer and is helpful in the planning of treatment.

Objective To explore the prevalence of ETS gene fusion in prostate cancer and its correlation with patient's clinicopathologic index.Methods Ninety-one samples from prostate needle biopsy cases with median age 75 (55-90) years and 18 samples from radical prostatectomy cases with median age 72 (63-81) years were collected from Oct.2010 to Feb.2012.TMPRSS2-ERG,TMPRSS2-ETV1 and TMPRSS2-ETV4 fusions were tested by multi-probe fluorescence in situ hybridization (FISH) assay.Fusion positive and negative cases were compared with age,TNM stage,Gleason score (median Gleason score 7 (6-9)),and PSA value (median PSA 33.4 μg/L,mcan PSA 118.8 μg/L).In needle biopsy cases,there were early stage 32 (34％),locally advanced prostate cancer 36 (40％),metastasis 23 (26％) ; in radical prostatectomy cases,there were 9 cases in T2 stage,8 cases in T3 stage,1 cases in T4a stage,respectively.Results TMPRSS2-ERG fusions were present in 14.3％ (13/91,95％ confidence interval,0.071-0.215)biopsy specimens and in 11.1％ (2/18,95％ confidence interval,0-0.256) radical prostatectomy specimens.No TMPRSS2-ETV1 or TMPRSS2-ETV4 fusions were found in any cases.Altogether,13 (86.7％)cases possessed deletion pattern.And 7 (46.6％) hold insertion pattern.5 cases had both deletion and insertion pattern.38.5％ (5/13) deletion pattern had distant metastasis.Except one metastatic case harbored both deletion and insertion pattern,there was no insertion pattern accompanied with metastasis.There were no differences between fusion positive and negative cases in the distribution of age (P =1.000),PSA (P =1.138),primary Gleason score (P=2.186),Gleason score (P=2.107),TNM stage (P=2.052) and Risk Degree (P=2.597).Conclusions The TMPRSS2-ERG fusion positive cases harbor more deletion pattern than insertion pattern.There are no differences between fusion positive and negative cases in the distribution of age,PSA,Gleason score,TNM stage and Risk Degree.

Objective To express the antigen gene Ts21 of Trichinella spiralis, purify the recombinant protein and test its antigenicity. Methods T. spiralis gene Ts21 was sub-cloned into the prokaryotic expression vector pMAL-c2X and the recombinant pMAL-c2X-Ts21 was constructed. The recombinant plasmid was transformed into E. coli TB1 strain and induced by IPTG. The expression products were purified by MBP-binding affinity chromatography. The antigenicity of the recombinant protein was examined by SDS-PAGE and Western blotting. Mice were immunized with the recombinant protein, the titer of the immune sera was detected by ELISA. The distribution of Ts21 protein in muscle larvae was observed by IFA. Results The molecular weight of the expressed fusion protein was about Mr 63 500 and the expression level peaked at 4 h post-incubation. The portion of the fusion protein accounted for 18.2% of all the protein by thin-layer gel optical scanning. Western blotting demonstrated that the recombinant protein was recognized by sera from mice infected by T. spiralis (T1) and T. nelsoni (T7) as well as sera of patients with trichinellosis, but not by sera from mice in-fected with T. nativa (T2), T. britovi (T3) and T. pseudospiralis (T4). The recombinant protein did not react with sera from patients with ancylostomiasis, cysticercosis and schistosomiasis, but cross-reacted with sera from patients with parag-onimiasis, clonorchiasis and echinococcosis. High titers of antibodies were produced in mice immunized with the recom-binant protein. IFA showed that the Ts21 protein was mainly distributed in the cuticle of muscle larvae. Conclusion The Ts21 antigen gene of T. spiralis has been expressed and the recombinant protein shows antigenicity.

Objective To investigate the effect of 5-azacytidine on cardiomyogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Method BMSCs were isolated from the marrow of adult SD rat’s femoral /tibial bones.Different concentration of 5-azacytidine were added to primary BMSCs on 3 d and cultured for different times.Cardiomyogenic differentiation of BMSCs was observed by immunohistochemistry and RT-PCR. Result After treated with 5-azacytidine, BMSCs proliferated slowly, became spindle-shaped after 10 d and aligned in a striated pattern after 20 d.TnT positive cells were showed by Immunohistochemistry and they expressed two cardiac-marked genes GATA-4 and ?-MHC.Thus 5-azacytidine induced cardiomyogenic differentiation of BMSCs in a time and concentration-dependent manner. Conclusion Our study suggests that bone marrow mesenchymal stem cells can differentiate into cardiomyocytes in vitro. They are ideal donor cells in cellular cardiomyoplasty for treatment of myocardial infarction.

Objective To establish a kind of detection technique of nucleic acid based on surface plasmon resonance(SPR) and to set up the foundation of real-time, online space microbial detection. Methods A portable online bio-molecules analyzer based on SPR biosensor was applied. The probe was mercapto-modified at the 5’ end and immobilized on the sensor surface. Then the target sequences in the solution were monitored and sensitivity, specificity and reproducibility of the method were investigated. Results The results showed that detection method with good specificity and sensitivity could realize online detection of target sequence. The system could detect 2.3 nmol/L target sequence, CV value of nine detections was 3.5% and that of thirty detections was 14.7%. Conclusion The established nucleic acid detection method has the advantage of high sensitivity, good specificity and reproducibility, which can be applied in the field of nucleic acid detection.

Objective To construct and express recombinant plasmid containing the structural gene encoding {Mr 31 000} antigen of Trichinella spiralis muscle larvae. MethodsThe target gene TspE1 was amplified by RT_PCR, cloned into pUC18 vector, and sub_cloned into the eukaryotic expression vector pcDNA3. BALB/c mice were immunized with the purified recombinant plasmid pcDNA3_TspE1 by gene_gun bombardment. The expression of recombinant plasmid in the skin tissue was observed by HE staining and immunohistochemical staining. Results and Conclusion The recombinant plasmid pcDNA3_TspE1 was successfully constructed and expressed in the BALB/c mice.

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are few in bone marrow, and they are easily mingled with other cells, especially red blood cells. Therefore, intervention needs to be depleted in the process of isolation of red blood cells so as to obtain highlypurified BMSCs as many as possible.OBJECTIVE: To isolate and culture BMSCs of rats with red blood cell lysis method, and perform biological identification.DESIGN: Observation and controlled trial.SETTING: Laboratory of Aerospace Cell Molecular Biology, Scientific Research Training Center for Chinese Astronauts.MATERIALS: Fifty 30-day-old male SD rats, weighing about 100 g, of SPF degree, were purchased from Beijing Experimental Animal Center (License No. SCXK (Jing) 2002-0003) and involved in this trial. DAB condensed chromogen (hydrogen dioxide included, Zhongshan Company), rabbit anti-ret polyclonal antibody (Boster Co.,Ltd., Wuhan), fetal calf serum (PAA, Austria) and LG-DMEM medium (Sigma Company, USA) were used in this trial.METHODS: This trial was carried out in the Laboratory of Aerospace Cell Molecular Biology, Scientific Research Training Center for Chinese Astronauts during September 2004 to September 2005. The rats were sacrificed by dislocation to expose bone marrow cavity. Cell suspension was collected. BMSCs were isolated and cultured primarily by whole bonemarrow culture method. ① Red blood cells lysis test: A, B, C and D 4 tubes were chosen and filled with 0.5 mL bonemarrow rinse solution which was filtered and fully beat upon. Then, red blood cell lysis buffer of 2 mL, ammonium chloride of 2 mL, phosphate buffer normal saline of 2 mL and 0.04 volume fraction acetic acid of 0.5 mL were correspondingly added into the 4 tubes. In each tube, absorbance and hemoglobin concentration were measured and cell growth was observed. ② Observation of growth curve, doubling time and surface marker molecule expression of BMSCs: Based on the formula, population doubling time (TD) =t[lg2/(lgNt-lgN0)] (NO and Nt represented the cell number after inoculation and t hours after culture ,respectively), cell population doubling time was calculated and traced and growth curve of the 2nd, 4th and 6th generations of BMSCs were analyzed; The proliferation of BMSCs was measured by methylene blue staining method; Surface marker molecule expression of BMSCs was detected with immunocytochemical staining.MAIN OUTCOME MEASURES: ①Observation of isolation and culture of bone marrowmesenchymal stems of rats. ②Effect of different methods on the lysis of red blood cells and the growth of BMSCs. ③ The growth curve and cell doubling time of the 2nd, 4th and 6th generations of cells. ④Surface marker molecule expression of BMSCs of rats.RESULTS: ① Results of isolation and culture of BMSCs of rats: After 48-hour primary culture, most of the cells had adhered to the wall, and 72 hours later, division growth of the adherent cells presented. Seven to eight days later, cell colonies formed obviously, and then increased quickly, expanded incessantly and fused with each other. On 14 to 16 days, cell clones grew densely. Immediately generative cells presented ball-shape, subsided and adhered the wall verysoon. Some few round cells suspended. Adherent cells distributed evenly and proliferated quickly within 3 to 5 days. Although cell morphology of generative cells did not change after passage, cell proliferation was speeded up obviously and cells covered the bottom of the whole bottom on about 6 days. ② Effect of different methods on red blood cell lysis and the growth of BMSCs: hemoglobin concentration in the red blood cell lysate-treated group, ammonium chloride-treated group and 4％ acetic acid-treated group was significantly higher than that in the phosphate buffer normal saline-treated group, with significant difference (P ＜ 0.01). ③ Observation of growth curve of different generations of BMSCs: The growth curves of the 2nd, 4th and 6th generations of the cells were basically the same: latent period about 1 to 2 days, then logarithmic growth phase, peak on the 5th day and finally plateau phase (about on the 5th to 7th days). The latent period of the 6th generation of BMSCs was not obvious and the population doubling time of BMSCs was about 34 hours. ④ldentification of immunophenotype of different generations of BMSCs: Both CD44 and CD106 staining of each generation of cells were positive, presenting brown granule sediments, and CD34 staining was negative.CONCLUSION: Inoculation of red blood cells lysate-treated bone marrow rinse solution can boost the adherent rate of BMSCs, and does not influence its post-adherent growth, so it is a feasible separation method. Cell surface marker staining confirms that thecells isolated in this study are BMSCs.

Objective:[WT5BZ] The effects of exogenous vitamin C (VC) and glutathione (GSH) on ultraweak spontaneous luminescence of culturing pulmonary alveolus macrophages from rabbits were studiesd. [WT5FZ]Methods:[WT5BZ] In a special thermostat, which was passed through by airs with various concentrations of oxygen, the alveolar macrophages (AMs) were cultured in DMEM medium with VC or GSH, and the spontaneous luminescence of culturing AMs was examined by a luminometer. [WT5FZ]Results:[WT5BZ] when VC in medium was over 0.3 mmol/L, it could significantly enhance the oxidative luminescence of cells exposed to O 2 and cell death was resulted. However, when the cells were exposed to air without O 2 there was no significant effect. On the contrary,the lower concentration of VC (0.03 mmol/L) as well as GSH could reduce the spontaneous luminescence of cells exposed to a high concentration (99.1%) of O 2 in air. [WT5FZ]Conclusion:[WT5BZ] The results show that the spontaneous oxidation of culturing cells is an important reason for the ultraweak luminescence. High concentration of VC can promote cellular oxidative damage in vitro, but the exogenous GSH has a protective effect against oxidization in culturing cells.

Objective To assess susceptibility of to 173 clinical M.tuberculosis isolates to by Micro-well Phage Replication Assay(MPRA).Methods To prepare the isolates and expose them to rifampin . To prepare phage D29 suspension . MPRA array for the susceptibility to rifampin.Results Compared with the absolute concentration method ,there were the same result of 38 isolate in 42 susceptible strains and of 124 isolate in 131 resistant strains. Between result of MPRA assay and absolute concentrstion (method,) concordance was 93.6%(38/42), susceptibility was 94.9%(124/131) and specificity is 90.5%((38/42).)Conclusion MPRA assay is a good and rapid method for drug susceptibility of rifampin.

Objective To observethe efficacy of mice infected with Sparganum mansoni by using different dosages of praziquantel.Methods A total of 156 Kunming mice were divided into 2 batches.each of them wag orally infted with 5 spargana.Thirty-six mice in the first batch were equally divided into 6 groups.the mice in group 1-5 were inoculated with spargana cultured in different concentrations of praziquantel for 3 days,the group 6 served as a control.One hundred and twenty mice in the second batch were equally divided into 12 groups,each mouse was inoculated with spargana obtained from frogs or tadpoles,group 1-9 were treated by different desages of praziquantel 1 or5 weeks post infection.group 10-12 served as controls.All of the mice wore sacriftced and dissectedl or 2 weeks after the treatment.the mean number of worms recovered was cmculated and worm reduction rates were determined.Results The number of worm recovered from mice infected with spargana cultured in 10-40 μg/ml of praziquantel had no significant difference with that of the control(P>0.05).The worm reduction rate wag 16.60%while the spargana beins cultured in 50 μg/ml of praziquantel.The worm reduction rates of the mice that sacrificed 1 week or 2 weeks after being treated by the same dosage of praziquantel had no significant difference(P>0.05).When being treated with 200.400 or 800 mg/kg of praziquantel 1 week post infection,the number of worm recovered from mice infected with spargana from frogs had no significant difference with those of the control 1 and 2 weeks after the treatment(P>0.05).The worm reduction rates between the groups with the same dosage 1 week and 2 weeks post treatment had no significant difference(P>0.05).When being treated with 200 or 400 mg/kg of praziquantel 1 week post infection,the number of worm recovered from mice infected with spargana from tadpoles had no statistically difference with that of the control 1 week after treatment (P>0. 05). The worm reduction rate of mice was only 17.02% while being treated with 800 mg/kg of praziquantel. The worm reduction rates among groups with different dosages had no significant difference (P>0.05). Compared with the mice infected with spargana from frogs treated with 1 200 or 1 800 mg/kg of praziquantel 5 weeks post infection, the difference between the numbers of worm recovered from mice 1 week and 2 weeks after treatment had no statistically significance (P > 0.05), but they were significantly higher than those of the controls (P<0.05 ). The worm reduction rates among the groups with the same dosage had no significant difference (P>0.05). ConclusionsPraziquantel (10-50 μg/ml) has no evident killing effect on spargnna in vitro, but when the dosage is higher(1 800 mg/kg), it has certain efficacy for treating the mice infected with spargana by oral inoculation.