Objective To study elimination of the Micro CT projective image noise and improve the quality of the recon-structed image for the Micro CT system.Methods The forming factors of the Micro CT projective image noise were analyzed,and proposed the Micro CT projective image correction method to correct the random noise,dark field,light field,response nonuniformity and defect picture element and eliminate the random noise and the ring artifact respectively.Results Through the comparative study,it proposes a simple and practical correction method for the actual response nonuniformity which combines the corrections of light field,dark current and gain inconsistency and settles the three corrections with one circu-lation.The method can enhance the algorithm efficiency greatly and the validity of the correction protocol can be proved through experiments.Conclusion The Micro CT projective image noise is eliminated effectively and the good basis for the following work are established such as three-dimensional reconstruction.

Objective To amend the errors between projective image center and the actual rotational center and improve the quality of reconstructed images. Methods The impact of reconstructed image in which the projection center and the actual center caused the deviation is analyzed, and provides the ways that is based on the gray value difference of objects and background in projection image to calculate actual rotational center, and then mends the projective image based on the actual rotational center. Results The difference between the actual rotational center and the calculated projective image center is less than one pixel. The quality of the reconstructed images is improved greatly. Conclusion The correction method is based on projective images, no need to increase the auxiliary conditions and the number of projection images. And the way as the pre-process of reconstruction, only needs small amount calculation, doesn't affect the reconstructive speed.

Objective To discuss some expectation on reform of classroom teaching method for eight-year program medical education, in order to provide a basis for the evaluation and reform of classroom teaching method. Methods Investigation that mainly included expectation on reform of classroom teaching method, classroom teaching method of most expected application, and ex-pected value of classroom teaching method was investigated and analyzed with self-made question-naire, to total 78 medical students (22 of Grade 2008, 20 of Grade 2009, 20 of Grade 2010, 16 of Grade 2011) of eight-year program who had completed the stage before medical education in the uni-versity, and related 32 teachers. Software of SPSS 20.0 was used to make statistical analysis. Results In the survey, 55.8% students and 78.1% teachers hoped to reform the current classroom teaching methods (χ2=4.782,P=0.029). 61%students and 90.6% teachers expected comprehensive application of diverse method(χ2=15.464,P=0.017), including instruction combined with writing on blackboard, teaching by multimedia, PBL, inversion-type class, independent learning, a lot of questions for exami-nation, and so on. The value expectation (>60%) of classroom teaching method was multivariate, in-cluding to stimulate students' interest in learning, to cultivate ability of finding and solving problems, conducive to understanding of knowledge and memory, conducive to comprehension and memory of knowledge, conducive to cultivation of learning method, able to improve ability of creativity and criti-cal thinking, conducive to strengthening interaction and communication of teachers and students, to improve the subject consciousness of student in classroom, helpful to cultivate the spirit of teamwork, to improve the efficiency of teaching, and able to make full use of information technology. Conclusions For the diversified value demand and the highly expectation on reform of current classroom teaching method, more attention should be paid to evaluation and guidance on reform of classroom teaching method for eight-year program medical education.

Objective To investigate the expression of Hippo pathway component in hepatic cancer tissues and investigate its effects on the tumor recurrence after Iiver transplantation.Methods The clinical data of 105 patients with liver cancer who were admitted to the Third Affiliated Hospital of Sun Yat-Sen University from July 2004 to September 2009 were retrospectively analyzed.The samples of liver cancer tissues were collected.The maximum diameter,number of foci,blood vessel involvement,preoperative level of alpha-fetoprotein (AFP),results of postoperative pathological examination were analyzed.All the patients were followed up via out-patient examination,mail and phone call.Patients were followed up once a week within the first month after operation,and once a month within the 6 months after operation,and then once every 3 months at 1 year later.The follow-up ended in December 2012 or tumor recurrence.The disease-free survival time began at the date of operation and ended at the time of tumor recurrence.The expressions of Yes-associated protein (YAP),phosphorylated YAP,Hippo pathway component (Lats1/2,pLats1/2,Mst1,pMst1/2) were detected by immunohistochemical staining.All data were analyzed using the chi-square test or Student t test.Factors might influence the postoperative tumorfree survival time after liver transplantation were analyzed using the Cox regression model.The survival curve was drawn by Kaplan-Meier method,and the disease-free survival was analyzed using Log-rank test.Results Positive expressions of YAP and phosphorylated YAP were detected in the nucleus and cytoplasm,and the positive expressions of Lats1/2,pLats1/2,Mst1 and pMst1/2 were detected in the cytoplasm.The positive expressions of YAP,phosphorylated YAP,Latsl/2,pLats1/2,Mst1 and pMst1/2 protein were 51.43％ (54/105),55.24％ (58/105),45.71％ (48/105),9.52％ (10/105),64.76％ (68/105) and 20.00％ (21/105),respectively.The positive expression of YAP was correlated with the tumor diameter,venous infiltration and AJCC stage (x2=4.173,9.611,7.233,P ＜ 0.05).The positive expression of Lats1/2 protein was correlated with tumor diameter and AJCC stages (x2=14.413,7.969,P ＜ 0.05).The positive expression of Mst1 protein was correlated with the tumor diameter (x2=4,129,P ＜0.05).The results of univariate analysis showed that the protein expressions of YAP,Lats1/2,pMst1/2,age,tumor diameter,tissue differentiation,preoperative level of AFP,venous infiltration and AJCC stages were risk factors influencing tumor recurrence after liver transplantation (HR =2.603,0.502,1.802,0.955,3.559,2.395,2.414,2.915,2.086,95％ CI:1.452-4.666,0.287-0.880,1.040-3.123,0.931-0.981,1.921-6.595,1.475-3.889,1.313-4.337,1.604-5.229,1.370-3.176,P ＜ 0.05).The results of multivariate analysis showed that the positive expression of YAP,tumor diameter ＞ 5 cm,low differentiation of tissue and AJCC stages Ⅲ were independent risk factors influencing tumor recurrence after liver transplantation (HR=2.011,2.176,2.390,1.574,95％CI:1.115-3.628,1.125-4.206,1.448-3.945,1.041-2.381,P ＜ 0.05).The median time of follow-up was 13.0 months (range,1.0-96.0 months).Eight patients missed follow-up.Fifty-four patients had tumor recurrence,and the mean time of tumor recurrence was 6.7 months (range,1.0-41.0 months).The disease-free survival time of patients with positive expression of YAP were significantly shorter than those with negative expression of YAP (Log-rank value =12.890,P ＜ 0.05).Conclusions Positive expressions of YAP and phosphorylated YAP were detected in the nucleus and cytoplasm,and the positive expressions of Lats1/2,pLats1/2,Mst1 and pMst1/2 were detected in the cytoplasm.The positive expression of YAP is the independent risk factor for tumor recurrence after liver transplantation.

Objective To preliminarily screen the neoplastic stem cell (NSC) related surface markers combination and to under-stand the NSC distribution in hepatoblastoma(HB) .Methods The children cases of HB undergone the surgical therapy in the Jian-gxi Provincial Children′s Hospital were selected .The immunohistochemical method was adopted for observing the expression and distribution of NSC-related markers CD34 ,Thy-1 ,c-kit ,CD56 and stem cell factor(SCF) in the HB tissue and the normal hepatic tissue away from the edge of tumor tissue 3cm outside .Results Thy-1 and c-kit were sporadically distributed in the HB tissue and mainly focued on the portal area ,but did not exressed in the normal liver tissue ;the expression of CD34 and SCF in HB was signifi-cantly higher than that in the normal liver tissue(P<0 .05) ,in which CD34 was mainly distributed in the endothelial system and SCF was mainly expressed in the portal area ;CD56 was expressed in the tissue of the clustered nerve fibers ,the expression differ-ence between them had no statistical significance (P>0 .05) .Conclusion The different NSC related surface markers are distributed in the HB tissue and focus on the specific areas .The positive cells of Thy-1/c-kit expression may play a role in the HB occurrence .

Objective To investigate the correlation between the Yes-associated protein (YAP) expression and epithelial-mesenchymal transition (EMT) by transiently transfecting with YAP small-interfering RNA (siRNA) in MHCC97H and MHCC97L cells.Methods MHCC97H and MHCC97L cells were transiently transfected by YAP siRNA.Furthermore,protein expressions and mRNA levels of characteristic markers of EMT (E-cadherin,N-cadherin) were examined by Western blotting and real-time polymerase chain reaction.Transwell invasion assay was used to detect changes of invasiveness of MHCC97H and MHCC97L cells.Results The YAP siRNA transfected group in MHCC97H was examined after 72 hours by Western blotting.The result showed obviously higher expression of E-cadherin in transfected group compared with the control group (P ＜ 0.05),and lower expression of N-cadherin (P ＜ 0.05).In MHCC97L cells,the expression of E-cadherin was also significantly increased (P ＜ 0.05),however,N-cadherin expression did not significantly change (P ＞ 0.05).Moreover,compared with the control group,transwell invasion assay showed that the number of the transfected group cells significantly decreased in MHCC97H(66 ± 6.89 vs 117 ± 7.23,P ＜ 0.05),and compared with the control group,the number of the transfected group also significantly decreased in MHCC97L (40 ±2.65 vs 77 ±4.33,P ＜0.05).The result of real-time polymerase chain reaction indicated that mRNA levels of E-cadherin increased (P ＜ 0.05),but mRNA levels of N-cadherin did not significantly change (P ＞ 0.05).This is considered as post-transcriptional regulation after silencing YAP in MHCC97H and MHCC97L.Conclusions YAP silencing is able to inhibit EMT in MHCC97H and MHCC97L cells by modulating the characteristic markers of EMT.The inhibition of YAP expression can reduce the invasion ability of hepatocellular carcinoma cells.

Objective To evaluate the effect of lipopolysaccharide (LPS) on the expression of ATP-binding cassette transporter A1 (ABCA1) in alveolar macrophage cells of rats.Methods The alveolar macrophage cells of rats NR8383 were seeded in 6-well plates at a density of 1 × 106 cells/ml (2 ml/well) and randomly divided into 6 groups:control group (group C,n =24),0.2 μg/L LPS group (group L0.2,n =12),2.0 μg/L LPS group (group L2.0,n =12),20.0 μg/L LPS group (group L20.0,n =60),200.0 pg/L LPS group (group L200.0,n =12),and 20.0 μg/L LPS + ABCA1 siRNA group (group L20.0 + siRNA,n =12).The cells were routinely cultured in group C.LPS with the final concentrations of 0.2,2.0,20.0 and 200.0μg/L was added to the culture medium in L0.2,L2.0,L20.0 and I200.0 groups,respectively.In group L20.0 + siRNA,siRNA 50 nmol/L was added to the culture medium and 12 h later LPS 20.0 μg/L was added.In group C,6 wells were chosen for determination of ABCA1 mRNA and protein expression.At 24 h of incubation with LPS 0.2,2.0 and 200.0 μg/L,or at 2,6,12 and 24 h of incubation with LPS 20.0 μg/L,6 wells were chosen and the cell suspension was obtained for measurement of ABCA1 mRNA expression (by real-time fluorescent quantitative PCR),and ABCA1 expression (by flow cytometry).At 12 h of incubation with 20.0 μg/L LPS or with 20.0 μg/L LPS + 50 nmol/L siRNA,6 wells were chosen and the cell suspension was obtained for measurement of TLR4 mRNA expression (by real-time fluorescent quantitative PCR) and TLR4 expression (by flow cytometry).Results Compared with group C,the expression of ABCA1 mRNA and protein was significantly down-regulated in L0.2,L2.0,L20.0 and L200.0 groups,and the expression of ABCA1 mRNA and protein was up-regulated in L20.0 and L20.0 + siRNA groups.In L20.0 group,the expression of ABCA1 mRNA and protein was gradually down-regulated with the prolonging time of incubation with LPS.Compared with group L20.0,the expression of TLR4 mRNA and protein was significantly up-regulated in group L20.0 + siRNA.Conclusion LPS can down-regulate the expression of ABCA1 in alveolar macrophage cells of rats,however,ABCA1 can inhibit the synthesis of TLR4.

Objective To find out related proteins that could possibly predict curative effects of neoadjuvant chemotherapy(NACT) by applying surface-enhanced laser desorption/ionization time-of-flight mass spectrometry(SELDI-TOF MS)to detect the difference in protein spectrum of breast cancer tissues.Methods Cancer tissue samples from 30 cases of breast cancer patients were detected by SELDI-TOF MS before NACT to get protein spectrum of breast cancer tissues.After NACT,the curative effects were evaluated according to RECIST standard and patients were divided into effective group(19 cases with complete response or partial response)and invalid group (11 cases with stable disease or progressive disease).Results 11 different protein peaks (P ＜ 0.05) were screened out:3491 Da(t =3.189 P =0.004),5158 Da(t =3.897 P =0.001),5360 Da(t =2.157 P =0.04),7549 Da(t =2.173 P=0.038),8451 Da(t =2.258 P =0.032),8694 Da(t =3.234 P =0.003),9089 Da (t =2.653 P =0.013),10528 Da(t =3.127 P =0.004),13445 Da(t =2.231 P =0.034),15118 Da(t =3.255 P =0.003),and 44065 Da(t =2.554 P =0.017).They were all expressed higher in effective group than in invalid group.The difference had statistical significance(P ＜ 0.05).Conclusion SELDI-TOF MS technology can screen out protein spectrum sensitive to NACT,providing evidences for breast cancer patients whether or not taking NACT.

Objective To explore ideal surgical axillary management of early breast cancer,and to determine the feasibility of reducing false negative rate of sentinel lymph node biopsy(SLNB)by combination of axillary suspicious lymph node biopsy and SLNB.Methods From Jan.2008 to Oct.2009,42 consecutive cases with early breast cancer were enrolled.All patients underwent suspicious node hook-wire location by doppler ultrasonography before operation.SLNB and suspicious lymph node biopsy were performed during operation.Complete axillary nodes dissection(ALND)or level II dissection would be conducted according to the biopsy result.The difference of node status prediction between SLNB and SLNB with axillary suspicious lymph node biopsy and was compared.Results All the 42 cases successfully underwent SLNB (100%).There were 2 false negatives occurred in SLNB,resulting in false negative rate of 11%,sensitivity of 88.9%and accuracy of 95.2%in predicting axillary nodes status.By contrast,SLNB with axillary suspicious node biopsy showed a false-negative rate of 0%,sensitivity of 100%,and accuracy of 100%.Conclusions Compared to SLNB in early breast cancer,combination of suspicious node biopsy and SLAB has a tendency of reducing false negative rate.However,Because of the limited samples,the difference has no statistical significance(P=0.2500).

Objective To investigate the effects of penehyclidine hydrochloride(PHC)precon-ditioning on hepatic ischemia-reperfusion injury (HIRI)in rats.Methods Fifty-four male Sprague-Dawley rats,weighing 230-250 g,were randomly divided into three groups (n =18 each):sham op-eration group (group S),HIRI group (group HR)and penehyclidine hydrochloride group (group PHC).In group S,the hepatoduodenal ligaments of rats were only pulled and separated,then abdo-mens were closed.In group HR,an atraumatic vascular clip was placed on the vessels blocking the portal venous blood supply to the median and left lateral lobes of the liver for 45 minutes,which re-sulted in approximately 70% rat liver ischemia injury.In group PHC,the rats were treated with 0.45 mg/kg penehyclidine hydrochloride at 30 minutes before the portal venous and hepatic arterial were blocked like group HR.Animals were killed at 2 h (T1 ,n =6),4 h (T2 ,n =6),24 h (T3 ,n =6) after HIRI or sham surgery.Liver tissues and blood samples were taken for analysis.The serum con-centration of ALT and AST were measured as the markers of hepatic functional damage.The TNF-a and IL-1βconcentration were measured by the enzyme linked immunosorbent assay (ELISA)tech-nique.The endothelial nitric oxide synthase (eNOS ) and hypoxia-inducible factor (HIF-1α) expression were measured by immunohistochemical staining.Results Compared with group S,the expressions of ALT,AST,TNF-αand IL-1βin groups HR and PHC increased at T1-T3 (P <0.05), which were similar to the change trend of HIR-1αand eNOS expressions (P <0.05).Compared with group HR,the expression of ALT,AST,TNF-αand IL-1βin group PHC decreased at T1-T3 (P <0.05).Nevertheless expression of HIF-1αand eNOS of groups HR and HPC were increased than that of group S,and experssion of HIF-1αand eNOS of group HPC were increased than that of group HR at T1 and T2 (P < 0.05 ).Conclusion PHC preconditioning can protect the liver from HIRI.The mechanism may be associated with the up-regulation of eNOS and HIF-1α,as well as reducing the in-flammatory response.