Abstract:Objective:To examine the expression and promoter CpG island methylation of glutathione S-transferase M3 (GSTM3) gene in clear cell renal cell carcinoma (ccRCC),and to evaluate the relationship of expression,methylation of GSTM3 gene with the metastasis and oncogenesis of ccRCC.Methods:Using semi-quantitative RT-PCR technique,we examined GSTM3 expression in surgical specimens of 24 primary ccRCCs and adjacent non-malignant tissues,14 metastatic ccRCCs and 2 ccRCC cell lines (RCC05-TXJ,RCC05-ZTJ) with different metastatic potentials.RCC05-TXJ cells were cultured in RPMI 1640 medium and treated with DNA methyltransferase inhibitor 5-aza-2′-deoxycytidine (5-Aza-CdR).Semi-quantitative RT-PCR was used to examine the expression of GSTM3 in response to 5-Aza-CdR treatment.Nested bisulfite sequencing PCR and DNA sequencing were used to analyze different methylation locuses in GSTM3 gene promoter in 10 primary ccRCCs and adjacent non-malignant tissues.We also examined the methylation level in 10 primary ccRCCs and the corresponding non-malignant tissues as well as 8 metastatic tissues by nested methylation-specific PCR.Results:Expression of GSTM3 gene in metastatic ccRCC cells (RCC05-TXJ) was lower than that in the non-metastatic ccRCC cells (RCC05-ZYJ).Down-regulation of GSTM3 gene expression was found in 17 of the 24 primary ccRCCs as compared with the non-malignant tissue.Expression of GSTM3 in the metastatic ccRCCs was lower than that in primary ccRCCs.5-Aza-CdR treatment increased GSTM3 expression in RCC05-ZYJ．Methylation in GSTM3 promoter was found in 4 of 10 ccRCC tissues,2 of 10 adjacent tissues,and 1 of 8 metastatic tissues.No significant difference was found between ccRCC and adjacent tissues (P=0.628) or ccRCC and metastatic disease (P=0.314) due to limit number of cases.Conclusion:Our findings support that promoter aberrant methylation is one of the major mechanisms of GSTM3 gene down-regulation in ccRCC.The preliminary identification of GSTM3 promoter methylation sites may provide a basis for further study.

Abstract:Objective:To investigate the methylation level of NPTX2 gene in pancreatic cancer and assess its clinical value in diagnosis of pancreatic cancer.Methods: We detected the mRNA expression and methylation levels of NPTX2 gene by qRT-PCR and quantitative methylation with SYBR Green in 10 primary pancreatic carcinoma tissues and the matched adjacent normal tissues.The methylation levels of NPTX2 gene were then detected and compared using the above-mentioned method in the whole blood of patients with pancreatic ductal adenocarcinoma and healthy volunteers.Results: The relative quantification (RQ) of NPTX2 expression in pancreatic carcinoma tissues was significantly lower than that in the adjacent tissues (0.276±0.263 vs 3.526±3.037,P＝0.001).The result of quantitive methylation indicated that the methylation index (MI) of the carcinoma tissues was significantly higher than that of the adjacent tissues (\[9.02±7.52\]% vs \[1.28±0.98\]%,P＝0.003).MI of NPTX2 gene was negatively correlated with RQ value (R=-0.552,P=0.012).We also found that the MI of the whole blood DNA methylation of patients with pancreatic cancer was significantly higher than that of healthy volunteers (\[1.80±1.76\]% vs \[0.84±0.45\]%,P<0.05).Conclusion: Our findings strongly suggest that NPTX2 gene is aberrantly hypermethyated in pancreatic ductal adenocarcinoma.Detection of MI of NPTX2 gene might be of great value for diagnosis of pancreatic cancer.

Abstract:Objective:To investigate the inhibitory effect of eukaryotic expression vector (attenuated salmonella typhimurium) carrying tumor necrosis factor-related apoptosis inducing ligand (TRAIL) and Chicken anemia virus VP3 gene on gastric cancer cells in vitro and in vivo.Methods: The cloning vectors pBud-TRAIL, pBud-VP3, and pBud-TRAIL-VP3 were transformed into attenuated Salmonella typhimurium by electric transformation technique. The S.typhimurium-based carriers were then transfected into gastric cancer cells. line SGC-7901 after stability assay. The expression of fusion green fluorescent protein was examined using fluorescent microscopy after 24 h. MTT assay was used to examine the inhibition of cell growth. Flow cytometry was used to detect cycle distribution and apoptosis rates of cells. The expression of caspase-3 and caspase-9 was assayed by immunohistochemistry method.Salmonella typhimurium carrying recombinant plasmid was administrated orally in sarcoma-bearing mice; 8 weeks later RT-PCR was used to detect the expression of cloning vectors in tumor tissue. Meanwhile, the sizes of tumors were also determined.Results: The recombinant plasmids were stably transformed into attenuated Salmonella typhimurium, and the plasmids was satisfactorily expressed in gastric cancer cells via attenuated Salmonella typhimurium. TRAIL and VP3 inhibited the proliferation of gastric cancer cells after 48 h. Flow cytometry analysis showed that the pBud-TRAIL-VP3 obviously enhanced apoptosis rates of gastric cancer cells. TRAIL and VP3 jointly increased the expression of caspase-3 and caspase-9. In vivo study showed that TRAIL and VP3 genes were expressed in tumor tissue and could inhibit the tumor growth（P<0.05）.Conclusion: Attenuated Salmonella typhimurium-mediated TRAIL and VP3 transfection of gastric cancer cells can inhibit cell growth in vitro and in vivo. The joint effect of TRAIL and VP3 is correlated with the increase of caspase-3 and caspase-9 expression.

Abstract:Objective:To investigate the possible association between codon 2020 polymorphism(Asn2020Ser) of IGF2R gene and the susceptibility to colorectal cancer(CRC) and hepatocellular carcinoma(HCC) in Chinese population. Methods: The IGF2R Asn2020Ser genotypes were determined by TaqMan assay in 345 CRC cases and 670 controls,and in 469 HCC cases and 558 controls. Odds ratios(OR) for cancer and 95% confidence intervals(CI) from unconditional logistic regression models were used to evaluate relative risks. Potential risk factors were included in the logistic regression models as covariates in the multivariate analyses on genotype and cancer risk. Results: The Ser allele of the IGF2R Asn2020Ser polymorphism was significantly associated with decreased risks of CRC and HCC. As compared with Asn/Asn genotype,Asn/Ser,Ser/Ser and Ser-allele carriers(Asn/Ser or Ser/Ser genotype) had significantly decreased risks of CRC,with the decrease being 0.71-fold (95% CI=0.54-0.94,P=0.017),0.64-fold(95% CI=0.42-0.97,P=0.036) and 0.69-fold(95% CI=0.53-0.90,P=0.008),respectively. A similar decreased HCC risk was also shown,with the decrease being 0.68-fold(95% CI=0.52-0.89,P=0.005) for Asn/Ser genotype,0.78-fold(95% CI=0.52-1.16,P=0.212) for Ser/Ser genotype,and 0.70-fold(95% CI=0.54-0.90,P=0.006) for Ser-allele carriers,when compared with Asn/Asn genotype. Conclusion: The Ser allele of the IGF2R Asn2020Ser polymorphism is potentially one of the protective factors against CRC and HCC in Chinese population.

Abstract:Objective:To investigate the expression of peroxisome proliferator-activate receptor γ(PPARγ) in the kidneys of rats at different ages,so as to study the possible association of PPARγ expression with oxidative stress in the aging process.Methods: Rat aging model was established using 3-,12-,and 24-month old rats.Western blotting was used to determining the protein expression of PPARγ.The changes anti-oxidative ability of renal tissue were studied by determining the activities of superoxidedismutase(SOD)and glutatio-neperoxidase(GSH-PX).Results: Western blotting assay showed that the renal expression of PPARγ protein decreased with the aging of rats,with that of 3-month group significantly higher than that of the 24-month group(P<0.01).The activities of SOD and GSH-PX deceased with the aging of rats(P<0.01).Both the SOD activities were positively correlated with the expression of PPARγ protein(r=0.90,r=0.89,P<0.01) Conclusion: Renal PPARγ has a decreasing tendency during the aging process of rats,which is correlated with the down-regulation of anti-oxidative ability of renal tissue.

Abstract:Objective:To examine the expression of Fas and Fas ligand(FasL) in cultured infantile hemangioma endothelial cells (IHECs).Methods: IHECs were isolated and cultured by explant combined with trypsin-digested techniques.Then the expression of Fas and FasL on cultured endothelial cells (ECs) were detected by flow cytometry (FCM) and fluorescent quantitative polymerase chain reaction (FQ-PCR) 2-ΔΔCT method,and the results in ECs were compared with those in Jurkat cells (positive control) and Human umbilical vein endothelial cells(HUVECs,negative control).Results: The positive rate of Fas in cultured IHECs was (90.97±2.36)% as detected by FCM,similar to that in the Jurkat cells (\[93.87±1.64\]%，P>0.05) and significantly higher than that in the HUVECs (P<0.01).Fas mRNA expression in the cultured IHECs was 1.260±0.721 as detected by FQ-PCR,and was 1.448±0.059 in Jurkat cells,with their ratio being 0.87 and with on significant difference found between the 2 group(P>0.05); the ratio of IHECs to HUVECs (0.354±0.170) was 3.56 and there was significant difference between the 2 groups (P<0.01).The expression of FasL mRNA in cultured IHECs,Jurkat cells and HUVECs was too low as detected by FCM and FQ-RCR to have any biological significance.Conclusion: Fas mRNA is highly expressed in cultured IHECs and FasL is rarely expressed in IHECs,which indicates that apoptosis of IHECs is associated with high expression of Fas in IHECs.

Abstract:Objective:To investigate the expression of chemokine receptor CXCR4 in hepatocellular carcinoma tissues,hepatocellular carcinoma cell line-MHCC97,human umbilical vein endothelial cells (HUVECs) and the ascites level of CXCL12,ligand of CXCR4,so as to lay a foundation for studying the role of CXCR4 in the metastasis of hepatocellular carcinoma.Methods: The expression of CXCR4 mRNA and protein was examined by RT-PCR and Western blotting in 21 specimens of hepatocellular carcinoma tissues,MHCC97 cells,HUVECs,and 17 specimens of normal hepatic tissues.Meanwhile,the levels of CXCL12 in ascitic fluids were assayed by ELISA in 18 hepatic cancer patients.Results: The relative expression values of CXCR4 mRNA in hepatocellular carcinoma tissues,MHCC97 cells,and HVECs were 2.21±1.09,2.14±1.15 and 1.72±1.20, respectively; and those of CXCR4 protein were 1.51±0.12,1.76±0.25, and 1.89±0.24,respectively; and those of CXCR4 protein were 1.51±0.12,1.76±0.25,and 1.89±0.24,respectively.CXCR4 mRNA and protein were not detected in normal hepatic tissues.ELISA results showed that the 18 hepatocellular carcinoma samples had a CXCL12 concentration range of 783-8 364 pg/ml (median value 6 871 pg/ml ) in ascitic fluids.Conclusion: CXCR4 is highly expressed in the hepatocellular carcinoma tissues and cells,which is not associated with the clinical staging of the cancer.The elevated CXCL12 level in the ascitic fluid of cancer patients indicate that CXCR4 may play an important role in the metastasis of hepatocellular carcinoma.

Abstract:Objective:To investigate the expression of DNA methylation transferase 1 (DNMT1) and histone deacetylase 1 (HDAC1) proteins in intraductal papillary mucinous neoplasms (IMPNs) and its possible clinical significance.Methods:The presence of DNMT1 and HDAC1 proteins in the 48 IPMNs and 54 pancreatic ductal adenocarcinomas (PDACs) were examined using SP immunohistochemical method and were statistically analyzed.Results:The expression levels of both DNMT1 and HDAC1 increased in a malignancy-dependent manner:normal pancreatic ductal →IPMA,IPMB → IPMC → PDAC.And both of them were positive in IPMNs of different subtypes.Conclusion:Higher expression of DNMT1 and HDAC1 is an early event of pancreatic malignancy.Expression of them can reflect the malignancy transformation of IPMNs,but can not be used for histological typing of IPMNs.

Abstract:Objective:To establish a real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) method for examining the expression of sialic acid-binding immunoglobulin-like lectin-one (Siglec-1)gene in the peripheral blood mononuclear cells (PBMCs) in patients with coronary heart disease (CHD),so as to explore the relationship between Siglec-1 mRNA expression and the development/progression of CHD.Methods: Based on the fluorescent SYBR Green method,a real-time quantitative RT-PCR was set up.The expression of Siglec-1 gene in the PBMCs of 57 patients with CHD and 38 healthy controls were measured by ABI PRISM 7000 Sequence Detection Systems.The blood lipid levels were determined in all the subjects by routine biochemical examination.Results: The mean Siglec-1 mRNA copy was significantly higher in the CHD group than that in the healthy control group (mean 3.23 folds,ranging 1.28-8.11 folds,P<0.01).The expression in the acute myocardial infarction group,stable angina and unstable angina group were 3.32(1.38-7.97),2.56(0.88-7.42),and 3.35(1.25-8.96) folds that of the control group,respectively.No significant difference was observed in the normalized Siglec-1 mRNA copy number between CHD group with normal level of serum lipids and abnormal level of serum lipids.Conclusion: A real-time quantitative RT-PCR method for detecting the expression of Siglec-1 gene in PBMCs has been successfully established.The expression of Siglec-1 is dramatically increased in PBMCs in CHD patients.

Abstract:Objective:To establish and identify androgen-independent human prostate cancer cell line LNCap by culturing LNCaP cells with gradual deprivation of hormone.Methods: LNCaP cells were cultured in the medium with gradual deprivation of hormone (treated by active carbon to simulate androgen deprivation)for 10 days; and then the cells were cultured with complete deprivation of androgen for 3 months till the cell entered the rapid proliferation phase again.The cell growth and expression of PSA and androgen were examined by CCK-8,immunfluorescence and RT-PCR methods.Results: LNCaP cells grew slowly after deprivation of hormone and took on a neuroendocrine phenotype and cluster growth pattern.After 3 months’ non-androgen culture,the cells regained original morphology and growth.CCK-8 indicated that LNCaP cells could grow in non-androgen condition; immunofluorescence assay indicated that LNCaP-AI cells could regain PSA-secreting activity in non-androgen condition; and RT-PCR suggested that androgen was highly expressed in LNCaP-AI cells.Conclusion: Androgen-independent LNCaP cell line can be established by culturing with gradual deprivation of hormone for 3 months.

Abstract:Objective:To establish a three dimensional finite element model of normal clavicle and calculate the stress and strain distribution on the clavicle under the axial load,so as to provide a numerical model for analysis of biomechanical behaviors.Methods: The right clavicle of a healthy normal male was scanned by CT and serial sections were obtained.Then the CT images were fed into the Simpleware and ANSYS software to create the model through a serial steps including meshing,assigning the material attribution,restricting the freedom and simulating axial loading.The result of stress distribution and response of the model was obtained.Results: The established model had 62 182 nodes and 40 878 tetrahedral units.The stress mainly concentrated within the middle 1/3 part of the clavicle under axial loading,and the stress on the inferior surface was obviously higher than that on the superior surface.Conclusion: The stress distribution analysis explains the reason why the middle 1/3 region of clavicle is prone to fracture.The stress mainly focuses on the inferior surface and the bending movement leading to clavicle fracture is located on the coronal plane,indicating that placement of plate in the superior surface can effectively balance the bending movement and avoid fixation failure during internal fixation of middle segment clavicle fracture.In a word,finite element analysis can be used to effectively investigate the biomechanic characteristics of clavicle.

Abstract:Objective:To rapidly construct bone-implant complex finite element model series using worksheet-driven CAD and finite element analysis software.Methods: The alveolar ridge and implant models were constructed by CAD software separately.Based on those models,the CAD models of different compositions of cancellous and compact bone and different implants were built and assembled.Electronic form software was utilized to drive the change of dimension in the process.All the models were imported into finite element analysis software and converted to finite element models after installation of different parts in the CAD software.Results: A total of 300 finite element models for different alveoli-implants were obtained through activating different versions of models.The models were of high quality and had no interference with each other.Conclusion: Our modeling method is rapid and highly effective.The series of models basically covers all types of alveoli,which provides a tool for studying the biomechanical characters of implants.

Abstract:Objective:To investigate the influence of the composite chitosan/heparin coated membrane through a layer-by-layer self-assemble technique on stent thrombosis.Methods: The composite chitosan/heparin coated membrane,which was assembled through a layer-by-layer technique,was attached to the inside surface of cylinder made from 316L stainless steel flake to form membranes of different layers of chitosan/heparin.The activated partial thromboplastin time (APTT),prothrombin time (PT) and thrombin time (TT) of the whole blood samples of healthy controls were examined by clotting method; and the stability of its anticoagulation effect was also examined.The influence of the prepared composite chitosan/heparin coated membrane on coronary stent thrombosis was investigated using swine model of arteriovenous shunt thrombus.Results: The APTT and TT were significantly longer in the composite coated membrane group compared with those of 316L stainless steel group (P<0.01).Moreover,the prolongation of APTT and TT was correlated with the increase of coated layers.The anticoagulant function of chitosan/heparin coated membrane was stable for two weeks(P>0.05).The weight of thrombus was significantly lower in the coated stent group than that in 316L stainless steel group(P<0.001).Conclusion: The composite chitosan /heparin coated membrane made by layer-by-layer self-assemble has effective and stable anti-thrombotic function.

Abstract:Objective:To observe the protective effect of cuttlebone polysaccharides (CBP-s) on gastric mucosa lesions induced by absolute ethanol in mice,and to study the related mechanism.Methods:Mice were pretreated by CBP-s for 3 or 5 days and then were administered with absolute ethanol orally to induce gastric mucosa lesions.Then the ulcer index,pH value of gastric contents,the levels of nitric oxide (NO),malondialdehyde (MDA),reduced glutathione (GSH),and the activity of SOD in the gastric mucosa were studied.Results:Mice gastric ulcer indices decreased after pretreated with 100 mg/kg,200 mg/kg,and 400 mg/kg CBP-s for 3 or 5 days.The pH values of gastric contents after 3 day-pretreatment were 3.09±0.20,3.32±0.15,3.38±0.15,and after 5 day-pretreatment were 3.43±0.23,3.34±0.25,3.68±0.24,respectively.MDA contents of gastric mucosa after 3 day-pretreatment with 400 mg/kg CBP-s and after 5 day-pretreatment with all 3 concentrations were (125.41±28.79),(145.80±26.13),(153.88±21.19),and (127.14±5.68) nmol/g,respectively,all significantly lower than that of the model group (P<0.05).GSH and NO contents were significantly increased in all but 100 mg/kg for 3 days group (GSH:\[282.93±32.33\],\[293.57±20.76\],\[260.79±39.23\],\[253.71±21.11\],\[359.38±10.89\] mg/g,P<0.05),(NO:\[23.40±4.41\],\[25.83±4.89\],\[21.60±3.05\],and \[22.75±2.44\] μmol/g,P<0.05).The activities of SOD were significantly increased in all groups (\[30.61±6.37\],\[45.35±1.90\],\[47.12±4.68\],\[36.48±5.64\],\[41.38±8.23\],and \[39.42±8.76\] U/mg,P<0.05).Conclusion:Our results indicate that CBP-s can protect ethanol-induced gastric mucosa from injury in mice.

Abstract:Objective:To investigate the expression of nuclear factor-κB (NF-κB) subunits P50 and c-Rel protein in primary cortical neurons of Wistar rats at different time points of oxygen glucose deprivation/reoxygenation(OGD/R).Methods: The neurons dissociated from the cortex of the neonatal rats were primary cultured and were identified by immunocytochemistry.OGD/R model was established.The study was divided into 6 groups according to different processing methods,including normal group,OGD 4 h treated,OGD 4 h/R 2 h treated,OGD 4 h/R 6 h treated,OGD 4 h/R 12 h treated and OGD 4 h/R 24 h treated groups.The expression of NF-κB P50 and c-Rel protein in neurons was examined by immunocytochemistry method and Western blotting.Results: (1) Immunocytochemistry detection targeting neuron specific enolase (NSE) and beta-Ⅲ tubulin confirmed that the cultured cells were neurons.(2)The expression of NF-κB P50 protein was significantly higher in OGD 4 h group than in control group(P<0.05); the expression continued to increase in OGD 4 h/R 2 h andOGD 4 h/R 6 h groups,and reached its peak 6 h after reoxygenation (P<0.01),then began to decrease,but the expression in OGD 4 h/R 12 h group was significantly different from that of the control group (P<0.05); there was no significant difference between OGD 4 h/R 24 h group and control group.(3) The expression of NF-κB c-Rel protein was similar between OGD 4 h group and the control group; the expression increased in OGD 4 h/R 2 h and OGD 4 h/R 12 h groups and reached its peak 12 h after reoxygenation(P<0.01),and did not recover to the normal level in OGD 4 h/R 24 h group (P<0.05).Conclusion: Oxygen glucose deprivation/reoxygenation can activate NF-κB subunits P50 and c-Rel in the primary cortical neurons in rats in a time-associated manner.

Abstract:Objective:To observe the effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) on the spatial learning ability/memory and dopaminergic neurons in the Nigra of senescence accelerated-prone 8 (SAMP8) mice.Methods: Three-month old male SAMP8 mice were injected with MPTP (36 mg/kg,s.c.) for 5 days,and animals in the control group were injected with NS (36 ml/kg,s.c.) in the same manner.Morris water maze was used to examine the searching strategy,seeking-platform latency,and the swimming time in the aimed quadrant.Immunohistochernistry was used to observe the changes of TH-ir positive neurons in substantia nigra.Results: The number of TH-ir neurons in substantia nigra pars compacta was significantly reduced in MPTP group compared with the control group(P<0.01).Morris water maze showed that the searching strategy of animals in MPTP group was worse than in the control group,with the seeking-platform latency of MPTP mice significantly prolonged (P<0.01),the time spent in the aimed quadrant significantly decreased (P<0.01) and time in the opposite quadrant significantly prolonged (P<0.05).Conclusion: MPTP can cause damage to the dopaminergic neurons in the substantia nigra of SAMP8 mice,which is subsequently followed by deficit in the spatial learning and memory in the animals.

Abstract:Objective:To investigate the effects of ischemic postconditioning and controlled low-pressure reperfusion on ischemia and reperfusion-induced spinal cord injury.Methods: Spinal cord ischemia was induced by occlusion of the descending thoracic aorta combined with maintaining systemic hypotension (40 mmHg,1 mmHg=0.133 kPa) in rats.Rats were randomly divided into three groups (n=103),namely,control group,postconditioning group (Post-con group) and low-pressure reperfusion group (LR group).Rats in control group underwent an abrupt increase to 100 mmHg in mean arterial pressure (MAP) after reperfusion.Those in the Post-con group underwent 3 cycles of 30-second reperfusion and 30-second occlusion after 30-second full reperfusion with MAP of 100 mmHg.The MAP maintained 40 mmHg for the initial 3 min of reperfusion in the animals of the LR group,then increased to 100 mmHg.The relative spinal cord blood flow (rSCBF) was monitored simultaneously during the ischemia and 30 min after reperfusion.The activities of superoxide dismutase (SOD),catalase (CAT) and myeloperoxidase (MPO) and malondialdehyde (MDA) content in lumbar spinal cord tissue were measured at 1,4,12 and 24 h after reperfusion,and the neurologic function of hind-limb was also evaluated at the same time.The samples of lumbar spinal cord were collected in all groups at the end of observation for histological examination.Results: Postconditioning and low-pressure reperfusion both improved the hyperperfusion after reperfusion.In Post-con group the activities of SOD and CAT was markedly increased 1 and 4 h after reperfusion (P<0.01) compared with control group,and no marked increase in MDA and MPO was observed.The MDA in the spinal tissue of LR group was significantly lower than that of the control group (P<0.01),but no significant increases were observed in the activities of SOD and CAT.The MPO activity in LR group peaked from the 1st to 4th hour of reperfusion and then gradually reduced.Postconditioning and low-pressure reperfusion both improved the neurologic function during reperfusion,but behavioral scores of animals in LR group were lower than that in the Post-con group.The pathological results had a similar pattern to the behavioral scores in all 3 groups 24 h after reperfusion.Conclusion: Ischemic postconditioning and controlled low-pressure reperfusion can both attenuate ischemia and reperfusion injury in spinal cord,and the neuroprotection of ischemic postconditioning is superior to controlled low-pressure reperfusion,probably because postconditioning can enhance activities of antioxidant enzymes and decrease neutrophil accumulation.

Abstract:Objective:To analyze the incidence of functional mitral regurgitation (FMR),post-operation characteristics and operation indications of Marfan patients with severe aortic regurgitation and giant left ventricle.Methods: The clinical data and the follow-up data of 19 Marfan patients with severe aortic regurgitation (≥10 ml) and giant left ventricle (LVEDd≥7.0 cm),who were treated from Jan.2000 to Dec.2007,were analyzed retrospectively.Results: All patients received Bentall operation and there was no perioperation death.Follow-up was 72 patient-year (ranging 3 months-7 years).The degree of FMR decreased obviously after operation,with an improvement rate of 87.5%,and the FMR almost disappeared 6 months to 1 year later.Conclusion: Bentall achieves good outcome in Marfan patients with severe aortic regurgitation and giant left ventricle.Mild to moderate FMR(<10 ml) disappears with the decrease of the left ventricle size.For patients with severe FMR (≥10 ml) complicated with dysfunction of left ventricle,managed ventricular pacing is suggested.

Abstract:Objective:To investigate the factors influencing the avascular necrosis of femoral head after internal fixation of femoral neck fracture.Methods: A total of 99 patients with femoral neck fracture,who were treated with internal fixation between May 1999 and April 2004,were followed up in outpatient Department and by telephone.The data was set up and analyzed by SPSS 14.0 software.Results: Until January 2006,avascular necrosis of the femoral head was found in 15 cases (15.2%).Age×internal fixation,gender×the time of reduction were found interactive in two factors interactive analysis.Age,the displacement of fractures,the quality of reduction,and internal fixation were found to be significant factors in single factor logistic regression analysis.Age,age×internal fixation,and the quality of reduction were found to be significant factors in multivariate logistic regression analysis.In fine analysis of interactive after multivariate logistic regression analysis,the removal of internal fixation was the most important risk factor for femoral head necrosis in 56-85yrs group,with the risk ratio being 26.00(95%CI=3.076-219.747).Conclusion: Elder patients have less chance for avascular necrosis of femoral head.The extraction of cannulated screws might lead to more avascular necrosis of femoral head and collapse in old patients.Dissection reduction can help to reduce avascular necrosis.

Abstract:Objective:To evaluate the clinical value of 64-slice spiral coronary CT angiography (CTA) in diagnosis of in-stent restenosis （ISR）after percutaneous coronary intervention(PCI) in patients with coronary artery disease.Methods: CTA was used to reconstruct and analyze the 345 segments (each stent was divided into three segments: proximal,middle and distal) of 115 stents in 60 patients with coronary heart disease after PCI.The results of selective coronary angiography (SCA) were taken as the golden standard to evaluate the sensitivity and specificity of CTA in diagnosis of ISR after PCI.Results: CTA clearly showed the location and length of the stents,the stenosis at stent and the characteristics of restenosis plaques.Thirty-nine in-stent restenosis lesions were found by CTA,including 8 calcified lesions and 31 non-calcified lesions.There were 25 lesions at the proximal end of the stent,7 at the middle and 7 at the distal end.Forty-two lesions were found by CTA,including 9 calcified lesions and 33 non-calcified lesions,with 26 at the proximal end,8 at the middle and 8 at the distal end.CTA correctly diagnosed 36 segments,missed 6,and misdiagnosed 3.The sensitivity,specificity,PPV,NPV and accuracy of CTA were 85.71%,99.01%,92.31%,98.04% and 97.39%, respectively.CTA had the highest sensitivity and specificity for diagnosis of the proximal ISR,being 96.15% and 100%,respectively.Proximal stent restenosis accounted for 64.10% of the total.Conclusion: CTA can clearly demonstrate the in-stent restenosis and has a high accuracy in diagnosing restenosis after PCI.CTA is a safe,simple and reliable noninvasive diagnostic method for diagnosis of in-stent restenosis after PCI.

Abstract:Objective:To analyze the risk factors,prevention and treatment of needle-tract bleeding after percutaneous image-guided radiofrequency ablation (PRFA) for hepatocellular carcinomas (HCC).Methods: A total of 1 567 RFA sessions were performed on 1 203 patients with 3 067 focuses of hepatocellular carcinoma between December 1999 and January of 2008.The clinical data of patients suffering from needle-tract bleeding were analyzed for risk factors,prevention,and treatment.Results: Seventeen (1.08%,17/1 567) patients developed needle-tract bleeding.Univariate analysis showed that the incidence of needle-tract bleeding was significantly correlated with the lower platelet count,prolongation of prothrombin time,the degree of liver cirrhosis,subcapsular HCCs and pattern of RF needle electrode (P=0.039; P=0.023; P=0.012; P=0.032 and P=0.016,respectively).Logistic regression model found that only lower platelet count,prolongation of prothrombin time,degree of liver cirrhosis and subcapsular HCCs were the independent risk factors for needle-tract bleeding after PRFA in patients with hepatocellular carcinoma (OR=4.032,P=0.040;OR=5.611,P=0.009;OR=3.871,P=0.046 and OR=5.216,P=0.026,respectively).Of the 17 patients 5 received only medicine therapy in the internal department,4 received surgery and 10 received transcatheter embolization.Thirteen patients survived and 4 died.Conclusion: It is indicated that prolongation of prothrombin time,lower platelet count,sub-capsular HCCs and pattern of RF needle electrode can influence the needle-tract bleeding after PRFA in hepatocellular carcinoma patients.The most effective methods to control the bleeding are transcatheter embolization and DSA or surgery for HCC patients after PRFA.

Abstract:Objective:To investigate the chemical constituents in the EtOAc fraction of the EtOH extract of Duchesnea indica Andr.Focke.Methods: Extraction was done with 95% EtOH.EtOAc fraction of the EtOH extract was isolated and purified by column chromatography using silica gel,RP-C18,Sephadex LH-20 and HPLC.The structures of the compounds were identified by their physico-chemical properties and spectral analysis.Results: Thirteen compounds were obtained from the EtOAc fraction of Duchesnea indica Andr.Focke and they were identified as euscaphic acid(1),arjunic acid(2),p-hydroxy cinnamic acid(3),apigenin(4),kaempferol(5),2α-hydroxy ursolic acid(6),2α-hydroxy oleanolic acid(7),tomentic acid(8),kaii-ichigeside F1(9),potengriffioside A(10),rosamultin(11),kaempferol-3-O-β-D-glucoside(12),and isoquercitrin(13).Conclusion: The compounds 2-5,10,and 12 have been isolated from the genus of Duchesnea for the first time.

Abstract:Objective:To compare the bioactivities of Taohua anti-diarrhea granula prepared by two different methods based on biothermo-kinetics study.Methods:The coarse and fine powder preparations of Taohua decoction were prepared.Microcalorimetry was used to obtain the metabolic curves and biothermo-kinetic parameters of Escherichia coli(E.coli) growth after treated by the two preparations to evaluate their bioactivities.Results:The values of growth rate constant k and maximum heat-out power Pm declined with the concentration increase of the 2 preparations; meanwhile,the values of peak time of the highest peak tm and growth inhibitory ratio I increased with the concentration increase of the two preparations.The results showed that the growth of E.coli was inhibited and the inhibitory effect was strengthened with the concentration increase of the two preparations.The IC50 was 35.9 mg/ml for the coarse powder preparation and 31.4 mg/ml for the fine powder preparation.Aggregate analysis of the metabolic curves and biothermo-kinetic parameters of E.coli indicated that the fine powder preparation of Taohua decoction had stronger inhibitory action against E.coli growth than the traditional coarse powder preparation.Conclusion:Microcalorimetry can be used to examine the bioactivity differences of the fine and coarse preparations of Taohua decoction.Superfine comminution may increase the inhibitory action of Taohua decoction.

Abstract:Objective:To analyze the service efficiencies of private hospitals in a Chinese city using Data Envelopment Analysis (DEA) method,and to discuss the problems existing in the management strategy of the hospitals.Methods: The management data of all the private hospitals were obtained by questionnaire survey and the parameters and samples were selected.DEA method was used to assess the service efficiencies of 58 private hospitals.Results: There were obvious differences on different aspects during their development between different hospitals.The total validities,the technical validities,and scale validities were obviously different.Twelve hospitals achieved total validity,23 achieved technical validity,and 14 achieved scale validity.Conclusion: Most of the private hospitals in the city have defects in scale and technical validities.In order to achieve total validity in service efficiency,the private hospitals have to pay more attention to balanced development of technique and scale,while considering their own situation.

Abstract:Objective:To study the antifungal activity of triazole alcohols by introduction of cyclopropyl as side chain. Methods: Ten title compounds were synthesized and characterized by 1HNMR,MS spectra and element analysis. The MICs of the compounds were determined by in vitro test in 8 fungus strains. Results: The title compounds exhibited potent antifungal activities against 8 strains,especially for the deep infection ones. Some compounds had MIC80 values less than 0.125 μg·ml-1 against Candida albicans,showing an activity 4 time higher than that of fluconzole.Conclusion: The title compounds with cyclopropyl and alky substituents have antifungal activities,and the antifungal activity decreases as the alkyl side chains getting longer.

Abstract:Objective:To understand the significance of serum TK1 level change during chemotherapy of patients with non-Hodgkin’s lymphoma.Methods: The serum TK1 levels were detected in 37 patients with non-Hodgkin’s lymphoma by enhanced chemiluminecence assay before chemotherapy and one day, four weeks after chemotherapy.Results: The changes of serum TK1 levels were significantly different at different stages of treatment (P<0.01).Conclusion: Our findings indicate that serum TK1 change might be of great value in evaluating the treatment outcomes and predicting recurrence of patients with non-Hodgkin’s lymphoma.

Abstract:As an antigen-presenting cell, dendritic cells (DCs)have drawn great attention due to its powerful antigen-presenting ability and immunological regulation function. Recently, it is believed that DCs also participate in regulation of immune response: DCs can not only stimulate immune response, but also induce immune tolerance. This heterogeneity of DCs determines that they play very important role in the immune response in tumors, autoimmunity diseases and graft rejections.

Abstract:Apoptotic mechanism of Pathogenic fungi has become a focus of study due to the worsening situation of clinical drug resistance in fungi.Apoptosis of Pathogenic fungi is a complicated network involving many pathways,regulators and effectors.Besides,it is closely related to the autophagy,mitochondrial apoptotic pathways,and changes of endogenous and exogenous factors.As a new target of antifungal agent,apoptotic pathways are of great clinical importance.This paper reviews the apoptotic pathways of Pathogenic fungi.

Abstract:Objective:To establish a sensitive and specific LC/MS/MS method for the determination of granisetron in human plasma,and to apply it in investigating the pharmacokinetics of granisetron hydrocloride capsule in healthy volunteers.Methods: Twelve healthy male volunteers were given single oral dose of 2 mg granisetron hydrocloride capsules.The plasma concentrations of granisetron were determined by LC/MS/MS method after liquid-liquid extraction.The pharmacokinetic parameters were calculated by noncompartment model.Results: The calibration curve was linear within a concentration range of 0.501-20.04 ng/ml,with the limit of quantification being 0.501 ng/ml.The main pharmacokinetic parameters of granisetron in the volunteers were as following: Cmax(6.93±1.90) ng/ml,tmax(1.35±0.29) h,t1/2(5.59±1.87) h,MRT (8.05±1.84) h,Cl/F(54.23±16.08) L/h,AUC0-24 (36.61± 8.87) ng·h·ml-1 and AUC0-∞ (39.32±9.36) ng·h·ml-1.Conclusion: The present method is accurate,sensitive and reproducible for the determination of granisetron levels in human plasma.It is suitable for pharmacokinetic study of granisetron hydrochloride capsule in human.

Abstract:目的:探讨PPARγ和PTEN在肾上腺皮质组织中的表达及相互关系。方法:应用免疫组化方法检测43例肾上腺皮质肿瘤(腺瘤30例、腺癌13例)及7例瘤旁正常组织标本中PPARγ及PTEN的表达，观察二者相互关系以及与肾上腺皮质病变病理学参数的联系。结果:肾上腺皮质癌与腺瘤和正常肾上腺组织比较，PPARγ表达明显增高(阳性率分别为76.9%、23.3%、0)，PTEN表达减低(阳性率分别为7.7%、43.3%、85.78%)；PPARγ表达与肿瘤大小、淋巴结转移相关，PTEN 与淋巴结转移相关(均P<0.05)；在肾上腺皮质肿瘤中PPARγ与PTEN表达存在负相关(P<0.05)。结论:PPARγ在肾上腺皮质肿瘤中高表达且与PTEN的表达显著相关，提示PPARγ与PTEN存在相互调控机制作用于肾上腺皮质肿瘤的发生发展，检测PPARγ及PTEN对肿瘤的早期诊断和良恶性鉴别有一定价值。

Abstract:目的:椎动脉V3段高度弯曲限制了后循环颅内支架通过的长度,术前如何准确选择支架的长度既能通过V3段又能良好地覆盖狭窄段血管是本文研究的目的。方法:术前对患者进行脑血管CTA检查,在血管重建过程中,恰好在一个平面内充分显示椎动脉V3段半环结构,按其最弯曲处的走行画一个圆环,此环与V3段半环结构内外缘平行,并位于两缘中间,过该环的中心测定其直径,代表半环结构的弯曲度。再沿最弯曲处内侧缘画一条切线,此切线与外缘相交于两点,测定此切线长。同时测定该处椎动脉直径。共对17例患者后循环颅内段存在21处狭窄以切线长作为支架长度的参考值选择支架,观察支架通过V3段的情况。另外,对男女各40例患者的CTA图像进行分析,结果用SPSS 15.0软件进行统计处理。结果:男女椎动脉V3段半环结构切线长与椎动脉直径、半环结构直径呈显著正相关（r＝0.44～0.92,P＜0.05～0.000)。男性左侧椎动脉直径及左侧半环结构直径明显大于女性（P<0.05,P=0.05）；而女性右侧半环结构切线长及右侧半环结构直径明显大于男性（P=0.05）。以切线长作为支架长度的参考值,17例患者手术中支架通过顺利,支架植入良好,手术顺利。结论:后循环颅内段动脉狭窄支架植入前,通过患者CTA图像直接测定V3段半环结构切线长度,作为支架长度的参考值,准确可靠,能使手术顺利开展。支架长度参考值（切线长）与椎动脉直径和半环结构直径呈显著正相关。

Abstract:目的:探讨室间隔缺损介入治疗术后发生传导阻滞的影响因素。方法:选择2006年1月至2007年10月间行介入治疗的室缺患者210例,男11 2例,女98例,年龄3～47(16±9）岁,全部应用国产封堵器。术前常规行12导联心电图检查,术后住院期间连续心电监测以及每天至少1次的12导联心电图检查。结果:210例室间隔缺损封堵术均获成功,其中41例术后7 d内发生了不同程度的传导阻滞,其中高度房室传导阻滞(AVB）6例,在术后21 d内恢复至术前,Ⅱ度ⅡAVB 2例,术后14 d内恢复正常,Ⅰ度 AVB 1例,术后7 d内恢复,束支传导阻滞32例,24例在7 d内恢复至术前,8例为新发束支阻滞未恢复。结论:心脏传导系统的损伤是室间隔缺损封堵术后常见的并发症,发生率较高,与患者的年龄、室缺的位置、大小、封堵器的类型、术中操作时间及术前心电图异常有关。

Abstract:目的:研究苯磺酸氨氯地平片在健康人体内的药物动力学和相对生物利用度,并与市售苯磺酸氨氯地平片进行生物等效性评价。方法:20例男性健康受试者双周期随机交叉单剂量口服10 mg苯磺酸氨氯地平片受试制剂和参比制剂,两次试验间隔为2周,采用LC/MS/MS法测定血浆中氨氯地平浓度。结果:受试制剂与参比制剂的血药浓度时间曲线基本一致,主要药代动力学参数Cmax分别为(9.72±0.90)和(9.57±0.96) ng/ml;tmax分别为(6.0±0.8)和(6.4±1.0) h;t1/2分别为(31.35±6.49)和(31.77±5.07) h;MRT 分别为(46.63±6.54)和(47.15±5.72) h;Cl/F 分别为(38.30±4.11)和(37.97±4.95) L/h;AUC0-96分别为(323.54±31.01)和(325.76±34.49) h·ng/ml。结论:统计分析结果表明两种制剂具有生物等效性,受试制剂的相对生物利用度为(99.86±13.48)%。

Abstract:目的:探讨Axin蛋白和MMP-14蛋白在食管鳞癌中的表达及与浸润转移的关系以及两者表达的相关性。方法:采用免疫组织化学(SP)法对72例食管鳞癌标本中Axin蛋白和MMP-14蛋白的表达进行了检测。结果:食管鳞癌中Axin蛋白的阳性表达率仅为38.9%,在有纤维膜浸润食管癌中的表达明显低于无纤维膜浸润者(P<0.05)；有淋巴结转移组的表达明显低于无淋巴结转移组(P<0.05)。食管鳞癌中MMP-14蛋白的阳性表达率为62.5%，在高分化的食管鳞癌组织中的表达明显低于中低分化者(P<0.05)；在有纤维膜浸润食管鳞癌中的表达明显高于无纤维膜浸润者(P<0.05)；有淋巴结转移组的表达明显高于无淋巴结转移组(P<0.01)。食管鳞癌Axin和MMP-14的表达具有相关性(P<0.05)。结论:Axin蛋白的低表达和MMP-14蛋白的高表达与食管鳞癌的浸润及转移均有关，二者表达呈负相关，有望成为判断食管鳞癌的转移潜能及预后的有效指标。

Abstract:目的:研究原发性病理性十二指肠胃反流患者胃黏膜组织中的凋亡细胞、增殖细胞，探讨原发性病理性十二指肠胃反流的发病机制。方法: 应用脱氧核糖核酸转移酶介导的缺口末端标记技术和免疫组化方法分析86例原发性病理性十二指肠胃反流和 42例正常对照胃黏膜细胞凋亡及增殖细胞核抗原的表达。结果:原发性病理性十二指肠胃反流组凋亡指数和增殖指数均显著高于正常对照组(P<0.05)，Hp阳性和Hp阴性亚组之间凋亡指数和增殖指数无显著性差异(P>0.05)。结论:胃黏膜细胞增殖与凋亡增强可能是原发性病理性十二指肠胃反流的发病机制之一，Hp感染未加重原发性病理性十二指肠胃反流所致胃黏膜细胞凋亡和增殖反应。

Abstract:补骨脂素(psoralen,PSO)是中药补骨脂中的主要成分。研究表明自中药补骨脂中提取的PSO加长波紫外线（UVA）,即PUVA对人白血病细胞NB4、K562、HL-60的生长均有抑制作用\[1-2\],并可诱导白血病细胞凋亡。本研究应用析因设计观察PUVA对NB4细胞线粒体膜电位（ΔΨm）的影响,以探讨PUVA诱导其凋亡的作用机制。

Abstract:磁性纳米颗粒除了具有一般纳米颗粒的特性外,还具有超顺磁性,在外加磁场作用下可定向操纵,因此其在生物医学中有着广泛的应用前景,如药物输送、磁共振成像(MRI)对比剂、靶向药物以及细胞分离和免疫分析等等\[1-3\]。超顺磁性氧化铁颗粒（superparamagnetic iron oxides,SPIOs）粒径小（直径不超过100 nm）,血液半衰期长（一般可达6 h）,是医学成像对比剂研究理想的纳米材料。其功能核团为Fe3O4晶体,急性毒理实验表明,短期内使用安全剂量的SPIOs对生物体无明显损害\[4-5\]。但是,大部分被网状内皮系统吞噬的纳米尺度Fe3O4晶体在体内的长期代谢过程尚未完全明了,在体内重要器官和组织内的分布亦不十分清楚\[6-8\]。鉴于其在生物医学领域的应用前景,本研究从组织器官水平研究该纳米材料对机体重要器官的形态和功能影响,通过单次和连续给药对其在体内的单次和重复积累作定量研究,为合理开发利用该纳米材料奠定实验基础。

Abstract:风湿性心脏病瓣膜置换术是我国目前最为常见的心脏瓣膜病手术,约占心脏瓣膜病手术的77%以上\[1\]。但如何进一步提高此类手术效果,减少术后并发症和病死率,尤其提高远期生存率,降低出血、栓塞等并发症的发生率,尚有待于进一步总结和探讨。我科自2001年1月至2006年6月行瓣膜置换术186例,现将手术体会和围手术期处理报告如下。