Abstract:Objective：To study the effect of cocaine- and amphetamine-regulated transcript (CART) protein vaccine on morphine analgesia and tolerance.Methods: The expression plasmid pGEX-4T3-CART was constructed by gene cloning.The CART protein was purified by glutathione s-transferase(GST)-affinity chromatography.The experiment included 6 groups:blank control,normal saline (NS),GST+Freund’s adjuvant,and CART protein vaccine (5 μg,10 μg and 20 μg) groups.After immunization for twice,all groups were tested in hot plate.Morphine analgesia effect was evaluated through s.c. injection with 6 mg/kg morphine solution,calculated by MPE%.Then morphine tolerance model was established,and the tolerance to morphine was tested by s.c. injection with 6 mg/kg morphine 12 h after the last injection.Results: CART vaccine itself had no pronounced effect on the pain threshold (P>0.05).CART vaccine at 10 μg significantly depressed the analgesic effect of morphine analgesia (P<0.05).Compared with NS group,vaccine groups showed a potential antagonizing tolerance effect,especially in the 10 μg group,with the MPE% significantly increased(P<0.05).Conclusion: CART vaccine itself has no influence on the pain response; however,it can impair the analgesia effect of morphine and can antagonize the analgesia tolerance to morphine.

Abstract:Objective：To investigate the modulatory effect of gonadal hormone on peripheral pain. Methods: Orchiectomized and ovariectomized rats models were established.Radio heater and Von-Frey hair were used to determine the peripheral mechanical and thermal pain threshold in gonadectomized rats and their corresponding sham-controls.Results: The body weight was increased and the uterus weight was decreased after ovariectomy in female rats（P<0.01）.There was no significant change in the body weight of male rats after orchiectomization.The 100% hind paw withdrawal threshold to Von-Frey hair stimulation was decreased significantly after ovariectomization in female rats,with no significant change in the thermal pain threshold.There was no significant changes in the 100% hind paw withdrawal threshold after Von-Frey hair stimulation or thermal pain threshold after orchidectomy in male rats.Conclusion: These results suggest that changes in androgen level of male rats have no influence on the peripheral basal pain threshold,and female gonadal hormone may inhibit the peripheral pain signaling and has no effect on thermal pain threshold.

Abstract:Objective：To investigate the inhibitory effects of L-type Ca2+-channel blocker nifedipine and magnesium sulfate on uterine contractility of full term pregnant women and its relation with labour.Methods: Myometrial strips,prepared from biopsies taken at caesarean section from labouring and non-labouring women at full term pregnancy,were mounted in organ chambers for isometric tension recording.Oxytocin(10-9mol/L) was employed to induce stable contraction.The cumulative effects of nifedipine(10-8,10-7,10-6 and 10-5mol/L) and magnesium sulfate (1,2,4,8 and 16 mmol/L) were studied by calculating the frequency,amplitude and AUC of contractions before and after drug applications,and the inhibitory rates were also calculated.Results: Nifedipine exerted a concentration-dependent relaxant effect on human term pregnant myometrial contractions induced by oxytocin.The relaxant effect was greater in the myometrium of non-labouring than that in the labouring group（P<0.05）.Magnesium sulfate also inhibited the contractility in both groups,with no significant difference between the 2 groups（P>0.05）.Conclusion: Decreased inhibition of human uterine contractility by L-type Ca2+-channel blocker nifedipine in term labouring myometrium may reflect sensitivity changes of Ca2+-channel to nifedipine after the onset of labour.The inhibitory effects of Mg2+ are not affected by labour,suggesting that Mg2+ may also exert its effect on contractility through other pathways besides Ca2+-channel.

Abstract:Objective：To study the damages within the coding region of the mitochondrial DNA in a human liver cancer cell line SMMC-7721 after exposure to different doses of X-ray irradiation.Methods: SMMC-7721 cells were radiated by different doses of X-ray (0,20,30,and 50 Gy).Primers for multiple PCR detection were designed according to the sequences of the coding region of human mitochondrial DNA,and the degree and location of mitochondrial DNA breaks were examined by LM-PCR (ligation mediated-PCR) and gene scanning technique.Results: The results of gene scanning showed the existence of X-ray sensitive breakpoints in the mitochondrial DNA coding region in human liver cancer cell line SMMC-7721,the degree of damage increased with the irradiation doses; the sensitive breakpoints were not distributed at random.The damage of heavy strand was severe than that in the light strand.Conclusion: X-ray irradiation can cause obvious damage to the coding region of the mitochondrial DNA in human liver cancer cell line SMMC-7721,and the changes of mitochondrial DNA may play a role in the clinical therapy of cancer.

Abstract:Objective：To express,purify and identify the protease (PR) of HIV-1 HXB2 subtype in E.coli,so as to screen the HIV-1 Gag CAP2/NC protein phage displayed library with randomized P2/NC protease cleavage sites and establish a phage model for in vitro screening of PR inhibitors. Methods: The primers were designed according to the PR amino acid sequence of HIV-1 HXB2 subtype and the E.coli preferred codon,the additional 5′-nucleotide sequence encoding the eight peptide MGTVSFNF for autocleave sites was inserted into the upstream of PR sequence. Then the PR107 DNA sequence was cloned into pET-32a vector which was used as expression vector in E.coli. Expression of HIV PR was induced by IPTG in E.coli BL21-DE3 and the expressed PR protein was purified by the Ni-NTA affinity column. The purified PR protein was refolded by diluted with MES buffer and blended into substrate protein CAP2NC to test its cleaving activity and the result was identified by SDS-PAGE. Results: The HIV PR107 DNA fragment with E.coli preferred codon was synthesized and was successfully inserted into the expression vector pET-32a. HIV-1 PR was expressed in E.coli BL21 DE3 after the induction by IPTG with a relative molecular weight of 30 000. The purified PR protein has a concentration of 2.54 mg/ml,and after refolded it could cleave substrate protein CAP2NC and this effect can be blocked by PI agent.Conclusion: PR107 DNA fragment with E.coli preferred codon of HIV-1 HXB2 subtype has been successfully synthesized and the PR protein has been successfully expressed,which can cleave substrate protein CAP2NC.

Abstract:Objective：To construct a mifepristone（an oral nontoxic chemical）-inducible eukaryotic expression vector and to evaluate its regulatory effect in vitro using luciferase reporter gene.Methods: Vector pDC-RULUC,which contains firefly luciferase reporter gene,promoter and mifepristone-inducible system,was constructed by molecular biological methods.A 1.2 kb insulator was inserted to reduce the interference between two transcription units.The vector was verified by PCR,restriction enzyme digestion,and sequencing.pDC-RULUC was used to transfect SW620 cells using Lipofectamine2000.Cells transfected with pGL3-Control and pGL3-Basic were used as positive and negative controls,respectively.Cotransfectant with pRL-TK renilla luciferase reporter vector was used as internal control.Cells of experimental group were incubated for 48 h in presence of different concentrations of mifepristone after transfection and were harvested for luciferase assay by using the Dual-Luciferase Reporter Assay System.Half of the wells were replaced with fresh medium and were measured after another 48 h.Results: The recombined plasmid vector was identified by digestion with different enzyme restrictions,PCR and sequencing analysis.The relative activity increased with the increase of mifepristone concentration.When the concentration of mifepristone reached 1×10-6 mol/L，the relative activity increased to approximately 50 folds of the original.No significant luciferase activity was detected when the mifepristone was removed.Conclusion:We have successfully established mifepristone-regulated eukaryotic expression vector, which can be used for controllable gene expression in vitro,providing a way for gene regulation and gene therapy.

Abstract:Objective： To establish a preoperative forecasting model for the nature of duodenal papillary tumors and to discuss its main predictors. Methods: A case-control study was conducted; the case group included patients with malignant duodenal papillary tumors and the control group included patients with benign duodenal papillary tumors. All the patients were from Changhai Hospital. Data of patients,including the demographic characteristics,clinical symptoms during onset,laboratory findings, and radiological data,were collected by face-to-face interviews or by reviewing the medical history. Chi-square,t-test or ANOVA were employed to performed univariate analysis. All factors with P values less than or equal to 0.25 in the univariate analysis were used as independent variables for multivariate analysis,and a Logistic regression forecasting model for the nature of duodenal papillary tumors was established. Results: Totally 199 patients with pathologically-confirmed duodenal papillary tumors were included in the present study,with 166 in the case group and 33 in the control group. Multivariate analysis showed that hemoglobin(Hb),total bilirubin(Tbil),direct bilirubin(Dbil),aspartate transferase(AST),alkaline phosphatase(AKP),gamma glutamyl transpeptidase(GGT),and carcinoembryonic antigen(CEA) were independent predictors for nature of duodenal papillary tumors,with their odds ratios(95% confidence interval) being 0.981(0.959,1.003),0.867(0.794,0.948),1.207(1.075,1.355),1.028(1.008,1.048),0.996(0.992,1.000),1.002(1.000,1.004),and 0.974(0.953,0.994),respectively. Conclusion: The Logistic regression model,which takes into consideration of Hb,Tbil,Dbil,AST,AKP,GGT,and CEA,can be used to predict the nature of duodenal papillary tumors,and its clinical value need to be further studied.

Abstract:Objective：To investigate the effect of carvedilol on expression of cardiac matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) after myocardial infarction in rats. Methods: An animal model of acute myocardial infarction (AMI) was established by descending left coronary artery ligation in 24 rats and they were divided into carvedilol (10 mg·kg-1·d-1) group (n=12) and normal saline group (n=12). Sham-operated group (n=9) received the same procedure but with no ligation. All animals were treated for 6 weeks via a gastric lavage. Heart function and hemodynamic parameters were determined after 6 weeks．The protein expression of cardiac MMP-2, MMP-9 and TIMP-2 was detected by immunohistochemical analysis in AMI groups, and the MMPs activities were assessed by zymography. Gene expression of myocardial MMPs/TIMPs (MMP-2,-9 and TIMP-1, -2) and cytokines (TNF-α, IL-1β) were measured by real-time quantitative PCR. Results: Compared with Sham-operated group，carvedilol group had significantly higher left ventricular end-diastolic pressure (LVEDP) and lower LV upstroke velocity (+dp/dtmax) and LV descent velocity (-dp/dtmax)（P<0.01）. Activities of MMP-2 and MMP-9, protein expression of MMP-2, MMP-9 and TIMP-2, and mRNA expression of MMP-2, MMP-9, TIMP-1, TIMP-2, IL-1β, and TNF-α were all higher in carvedilol group compared with sham-operated group (P<0.05). Compared with normal saline group, carvedilol group had lower LVEDP（P<0.01）, higher +dp/dtmax, -dp/dtmax(P<0.05),lower activities of MMP-2, MMP-9（P<0.01）, lower protein expression of MMP-2, MMP-9 and TIMP-2, and lower mRNA expression of MMP-2, MMP-9, TIMP-2, IL-1β, TIMP-1, and TNF-α (MMP-9 P<0.01，others P<0.05). Conclusion: Carvedilol can obviously decrease cardiac expression of MMP-2 and MMP-9 and slightly decrease expression of TIMPs; it can also decrease secretion of MMPs through decreasing IL-1β and TNF-α expression,thus prevents myocardial extracellular matrix remodeling,reverses ventricular remodeling,and subsequently improves cardiac function.

Abstract:Objective：To characterize the anatomic morphology of pulmonary vein ostium by 2-D and 3-D computed tomography angiography.Methods: Thirty patients with paroxysmal/persistent atrial fibrillation (AF) undergoing computed tomography angiography before catheter ablation of AF were analyzed for the anatomic morphology of pulmonary vein ostium in 2-D and 3-D manner.Results: The diameter in axial,coronal and sagittal views of diameters of the pulmonary veins ostium (PVs) were (16.90±4.79) mm,(21.37±4.23) mm,(22.41±3.96) mm in the left superior pulmonary vein(LSPV),(13.50±3.99) mm,(15.84±3.22) mm,(16.82±3.63) mm in the left inferior pulmonary vein(LIPV),(17.77±4.69) mm,(19.11±4.10) mm,(19.71±4.33) mm in the right superior pulmonary vein(RSPV),and (15.33±3.88) mm,(16.20±4.00) mm,(17.10±4.24) mm in the right inferior pulmonary vein(RIPV).The maximal and minimal diameters (dmax and dmin) of PVs in 3-D view were (24.30±4.54) mm,(17.76±4.24) mm in LSPV,(19.10±4.45) mm,(12.27±3.52) mm in LIPV,(22.99±5.04) mm,(16.19±4.87) mm in RSPV,and (18.63±4.60) mm,(14.46±3.48) mm in RIPV.There is difference of diameters between SPVs and IPVs （P<0.01）in two-dimensional and three-dimensional view except for RIPV. Conclusion: CT imaging can present precise 3-D reconstruction of the pulmonary vein ostium,allowing for understanding the details before catheter ablation.There is great variance in the dimension of the pulmonary vein ostium among individuals,which should be taken into consideration before operation.

Abstract:Objective：To observe the changes of left ventricular (LV) systolic strain(ε) and strain rate(SR) in patients with hypertrophic cardiomyopathy (HCM) and normal healthy subjects by velocity vector imaging(VVI).Methods: Twenty patients with HCM and 20 age-matched healthy adults were enrolled in this study. The peak systolic ε , SR of 18 segments in the 4, 3 and 2-chamber views were measured by VVI. Results: It was found that the systolic SR was (-1.06±0.09)/s in the healthy control group and (-0.74±0.16)/s in the HCM group（P<0.05）. The reduction of systolic SR in HCM group was found not only at the hypertrophic regions but also at normal regions. Conclusion: The systolic ε and SR in HCM group is lower than that in the normal control group. The reduction of systolic ε and SR in HCM group is not limited to the hypertrophic region.

Abstract:

Abstract:Objective：To study the effect of GnRH analog triptorelin in resensitizing cisplatin-resistant human ovarian cancer cells and to discuss the related mechanism. Methods: Cisplatin-resistant human ovarian cancer cell line OVCAR-3/CDDP was established in vitro. MTT assay was used to assess the inhibitory effects of triptorelin, cisplatin alone or a combination of both on OVCAR-3/CDDP cells. Flow cytometry was employed to observe the expression changes in epidermal growth factor receptor (EGFR) in different groups. Results: The drug restant index of OVCAR-3/CDDP cells was 13.42. The resensitizing fold of cisplatin combined with triptorelin was 3.80. The expression of EGFR had the most prominent decrease in OVCAR-3/CDDP cells in the combination group. Conclusion: Triptorelin can partially resensitize cisplatin-resistant OVCAR-3/CDDP cells, which might be related to the down-regulation of EGFR.

Abstract:Objective：To observe the anti-tumor activity of oxymatrine alone or in combination with cyclophosphamide against Lewis lung carcinoma in mice and to discuss the related mechanisms. Methods: Lewis lung carcinoma cells were transplanted subcutaneously to establish cancer transplantation model in mice. Animals were orally given oxymatrine(80, 40 and 20 mg/kg) alone or in combination with cyclophosphamide(30 mg/kg, gastric lavage). The mean tumor inhibitory rate was calculated and the growth curve of tumors was plotted. Radio immunoassay(RIA) was used to determine the serum concentration of epidermal growth factor (EGF). Comparison was made between the treatment groups and the control group. Results: The inhibitory rates were not significantly different between the control group and the pure oxymatrine treatment group. When combined with cyclophosphamide, the inhibitory rates of oxymatrine were significantly increased (P＜0.01). When at 80, 40 and 20 mg/kg, oxymatrine inhibited tumor growth by 66.62%, 61.70% and 59.04%, respectively. Also, when combined with cyclophosphamide, oxymatrine at 40 and 80 mg/kg significantly decreased EGF levels compared with those of model group and pure oxymatrine groups (P＜0.01). Conclusion: Oxymatrine can enhance the inhibitory effect of cyclophosphamide against Lewis lung carcinoma, and the mechanism may be related to the decrease of serum EGF level.

Abstract:Objective：To explore the expression of nerve growth factor (NGF), its receptor P75NGFR and TrkA in the human pancreatic cancer tissues, and to study the relationship between the expression and clinical-pathological indices.Methods:The expression and morphological distribution of NGF, P75NGFR and TrkA were investgated in 65 pancreatic cancer tissues and 18 normal pancreas and chronic pancreatitis tissues by means of immunohistochemistry; the relationship between the expression and clinical-pathological indices was also investigated.Results:The expression of NGF and TrkA in human pancreatic cancer tissues was higher than that in the normal pancreas and chronic pancreatitis.Expression of P75NGFR had no significant change in the two groups.The expression of NGF was positively correlated with tumor size and was also associated with lymphatic invasion.The expression of P75NGFR and TrkA was closely correlated with lymphatic invasion.We also found that the expression of NGF was correlated with histological differentiation, tumor stage, tumor size and lymphatic invasion; the closest relationship was found between the expression of NGF and tumor size.The expression of P75NGFR was correlated with lymphatic invasion,the expression of TrkA was correlated with both lymphatic invasion and tumor TNM stage.Patient’s prognosis after surgery was mainly correlated with lymphatic invasion and the positive expression of TrkA.Conclusion:NGF, P75NGFR and TrkA participate in the progress of pancreatic cancer.TrkA is closely related to pancreatic cancer and can serve as an important target for gene therapy of pancreatic cancer in the future.

Abstract:Objective：To investigate the concentrations of secreted protein acidic and rich in cysteine(SPARC) in the serum and urine of patients with IgA nephropathy and its expression in the kidney tissues.Methods: The concentrations of SPARC,tumor necrosis factor-α (TNF-α),interleukin 1β (IL-1β),and interleukin 6 (IL-6) in the serum and urine were measured with enzyme-linked immunosorbent assay（ELISA）.The contents of SPARC protein in the culture medium of human mesangial cell (HMC) and human renal tubular epithelial cell (HKC),which had been treated with IL-6,were determined by ELISA.The expression and distribution of SPARC in IgA nephropathy and normal kidney tissues were observed by immunohistochemistry assay.Results: The concentrations of SPARC in serum and urine of IgA nephropathy patients were higher than those of the normal control subjects ( \[2.43±1.22\] μg/ml vs \[0.69±0.21\] μg/ml,\[7.73±2.81\] μg/ml vs \[1.17±1.03\] μg/ml,P<0.01).The serum levels of TNF-α,IL-1β and IL-6 in IgA nephropathy group were significantly higher than those in the control group (P<0.05); the urinary levels of TNF-α and IL-6 in IgA nephropathy group were also higher than those in the controls (P<0.01).SPARC protein secreted by HKC was higher than that by HMC (P<0.01).SPARC was weakly positive in normal distal cortical tubules.SPARC protein expression in tubular epithelial cells of IgA nephropathy patients was obviously higher than that of the normal controls. Conclusion: The secretion of SPARC by renal tubular epithelial cells is increased in patients with IgA nephropathy,which results in elevation of serum SPARC and may have a protective feedback inhibitory effect on HMC proliferation.

Abstract:Objective：To explore the relationship of the local recurrence with the expression of protein P53 and PCNA in the primary lesions and the surgical margins of laryngeal carcinoma.Methods:The primary lesions and the surgical margins of laryngeal carcinoma of 36 patients were made into serial sections.Immunochemical method (H-E staining) was used to detect the expression of P53 and PCNA protein.Results:The positive rates of protein P53 and PCNA were 55.6% (20/36) and 88.9% (32/36) in the primary lesions,and 25%（9/36） and 30.6%（11/36） in the surgical margins,respectively.The recurrent rates of the primary lesions positive for P53 and PCNA were 35%(7/20) and 25%(8/32),respectively,which were higher than those negative for them (12.5%\[2/16\] and 25%\[1/4\]).The recurrent rates of surgical margins positive for p53 and PCNA were 44.4% (4/9) and 72.7% (8/11),respectively,which were higher than those negative for them (18.5%\[5/27\] and 4%\[1/25\]).The expression of P53 and PCNA proteins in the primary lesions and surgical margins was abnormal; the recurrent rate of the laryngeal carcinoma was 100% (4/4) after surgery.Conclusion:P53 and PCNA protein can be used as biomarkers for local recurrence of laryngeal carcinoma after operation.Detection of P53 and PCNA in both the primary lesions and the surgical margins may be of greater significance in forecasting local recurrence.

Abstract:Objective：To investigate the pathologic characteristics of lymphangiogenesis in renal transplants and to analyze its clinical implication.Methods: The morphology and distribution of lymphangiogenesis were investigated by a biotin-streptavidin horseradish-peroxidase method with anti-podoplanin monoclonal antibody in 45 archival biopsies.The lymphatic vessel density (LVD) was calculated and the results were compared between different pathologic types and with the normal renal tissues.Results: Fewer podoplanin-positive lymphatic vessels were identified in the biopsies from the renal transplants with normal function,and the transplants had a similar morphological profile as normal renal tissues.More podoplanin-positive lymphatic vessels were observed in the transplants suffering acute rejection episode; the vessels mainly located around peripheral arteriole with different lumen sizes.Transplants with chronic rejection had the most podoplanin-positive lymphatic vessels with focal mononuclear infiltration and distended/distorted lymphatic vessels.The lowest mean LVD (1.26±0.27) was observed in the biopsies from transplants with normal function,which was significantly different from those in the acute rejection and chronic rejection groups (P<0.05).The highest mean LVD was found in the chronic rejection group (20.76±5.30),which was significantly higher than those of the other 2 groups (P<0.01); no significant difference was observed between the transplants with normal function and the normal kidney.Conclusion: Lymphatic neoangiogenesis occurs in the renal transplant and its pathologic characteristics differs in the transplants with different rejection types.

Abstract:Objective：To observe the expression of osteopontin (OPN) in the giant cell tumor of bone (GCTB) and to discuss its clinical significance.Methods: OPN expression was examined in 77 GCTB samples by EnVisionTM two-step immunohistochemical method. And the expression was compared between patients with different clinical and pathological data; the implied clinical relevance was also analyzed. Results: The positive expression rate of OPN in the GCTB was 76.6%; only weak expression was noted in the normal bone tissues. The expression in the invasive GCTB was significantly higher than that in the non-invasive ones（95.5% vs 69.1%, P=0.013）. The expression rate of OPN increased with the increase of Campanacci stages of GCTB, and significant difference was found between different stages（P=0.019）. The recurrence status, gender, age, surgery types, duration of illness or site of tumor were not associated with the positive rate of OPN. Conclusion: OPN is highly expressed in GCTB tissues, and the expression is associated with the invasiveness and the Campanacci stages of GCTB.

Abstract:Objective： To summarize our experience in perioperative anesthetic management for fulminant hepatic failure (FHF) patients receiving liver transplantation.Methods:The clinical anesthetic data of 48 FHF patients receiving orthotopic liver transplantations (OLT) from January 2006 to January 2007 were retrospectively analyzed,and the anesthetic management experience was summarized.General anesthesia was applied; the hemodynamics was monitored during the operation and doses of adrenaline and phenylephrine were adjusted according to the monitoring results. Blood samples were obtained before operation,before anheptic,30 min after anhepatic phase,5 min before neohepatic phase,and 5 min,30 min and 60 min after neohepatic phase for blood gas and electrolyte analysis and for determination of coagulation function; the drugs were subsequently adjusted according to analysis results.Results:All the 48 patient underwent successful anesthetic management and there was no death during opearation.The average blood loss during operation was (5 219±478) ml.Mild alkalosis,hypokalemia,hyponatrium,and hypocalcemia were present before operations.pH,BE and HCO3- were obviously reduced 30 min after anhepatic phase and increased 60 min after neohepatic phase.Kalemia was obviously increased 30 min following anhepatic phase and began to increase 60 min following neohepatic phase.Calium concentration was decreased at the end of preanhepatic phase (P<0.05).Prothrombin time (PT) before operation was significantly longer than the norm (P<0.05) and tended to recover after treatment; however,it prolonged again 5 min before and after neohepatic phase.The fibrinogen (FIB) level was significantly decreased compared with that before operation (P<0.05) and gradually recovered at the end of the operation.There was no rapid decrease in the cardiac output or mean aterial pressure at anhepatic phase,and the hemodynamics was stable at other phases.Conclusion:FHF patients are often complicated with disorders in the inner environment; their hemodynamics,blood gas and coagulation functions should be monitored perioperatively,and drugs should be adusted accordingly to maintain the stability of the inner environment.

Abstract:Objective:To study the effect of total saponins on cerebral blood flow and vascular resistance in anesthetized dogs.Methods: Thirty hybrid dogs in either sex,with a body weight of (11±1.5) kg,were evenly randomized into 5 groups:negative control group(saline 5 ml/kg,ig),positive control (nimodipine 300 μg/kg,iv),and 3 groups treated with total saponins (low-dose group \[10 mg/kg,ig\],middle-dose group\[30 mg/kg,ig\],and high-dose group\[60 mg/kg,ig\]).The dogs were anesthetized with intravenous pentobarbital sodium (30 mg/kg).The right common carotid artery was exposed to measure the cerebral blood flow,cerebral vascular resistance,blood pressure and heart rate using the MFV-3200 electromagnetic flow meter and MPA-3000 bioelectricity signal-amplifier.Results: Compared with negative control, cerebral blood flow was significantly increased in animals treated with asparagus root saponins（30 and 60 mg/kg,ig）during 5 and 120 min after drug administration (P<0.01). No significant effect on cerebral blood flow and vascular resistance was found in animals treated with asparagus root saponins（10 mg/kg）. Conclusion: Asparagus root saponins can increase cerebral blood flow in anesthetized dogs. \[KEY WORDS\]total saponins from Asparagus cochinchinensis; anesthesia; cerebral blood flow; vascular resistance; blood pressure; heart rate

Abstract:Animal models of pancreatic cancer are important for the experiments of pancreatic cancer research.An ideal animal model of pancreatic cancer provides effective tool for exploring the tumorigenesis of pancreatic cancer.This paper summarizes the methods for establishing mouse model of pancreatic ductal adenocarcinoma and discusses interpretd the advantages and disadvantages of different models.

Abstract:Recently,study on cencer stem cells has been a focus of study.Cancer stem cell is a small population of cencer cells possessing the properties of stem cells：self-renewal,differentiation and proliferation.To date,the existence of cancer stem cells has been proven in acute and chronic myeloid leukemia,breast cancer,brain tumors,liver cancer and colon cancer，etc..In this article we reviews the current progress on cancer stem cells，including the defination,existing evidence,research methods,and challenges in clinical application.

Abstract:特发性炎性肌病（IIM）是一组由自身免疫介导的弥漫性结缔组织病。其发病机制未明，可能与免疫异常有关。肌细胞上主要组织相容性抗原复合物的过度表达可能是肌炎的启动因素，而肌炎的持续与发展则与机体免疫应答异常有关。皮肌炎以体液免疫损伤为主，而多发性肌炎则以细胞免疫损伤为主。各种细胞因子、趋化因子和黏附分子在IIM的免疫病理过程中发挥着重要作用。

Abstract:目的：观察皮肌炎(DM) 患者外周血B淋巴细胞刺激因子(BLyS)的含量及其mRNA在外周血单个核细胞（PBMCs）中的表达。方法：ELISA法测定10例皮肌炎患者外周血中BLyS含量,同时采用RT-PCR技术检测BLyS mRNA在PBMCs中的表达情况,并取健康体检者作对照。结果：皮肌炎患者外周血BLyS 为(10.25±1.19) μg/L,显著高于健康对照组\[(3.19±0.65) μg/L；t=5.186,P<0.01\]；BLyS mRNA在皮肌炎患者外周血单个核细胞中高表达(0.39±0.04)%,明显高于健康对照组\[(0.17±0.01)%; t=5.611,P<0.01\]。结论：皮肌炎患者外周血中BLyS含量较正常人明显增高,同时伴有PBMCs BLyS mRNA的高表达,提示BLyS分子水平的改变与皮肌炎的发生发展有着密切关系。

Abstract:目的：观察血清同型半胱氨酸及白细胞介素-6、超敏C反应蛋白在不同病情程度急性格林-巴利综合征病程中的变化并阐明其临床意义。方法:110例急性格林-巴利综合征患者，选择其中病情重者35例作为观察组1，一般情况相似且病情中等者35例作为观察组2，一般情况相似病情轻者40例作为观察组3，另设健康查体者50例作为对照组。分别测定发病后1、14、28 d血清同型半胱氨酸及白细胞介素-6、超敏C反应蛋白浓度。结果:发病后1 d观察组1、2、3血清同型半胱氨酸及白细胞介素-6、超敏C反应蛋白浓度均明显高于对照组（P<0.01），且各观察组之间比较无显著性差异；发病后14 d观察组1、2上述指标明显高于观察组3及对照组，观察组3与对照组之间比较无显著性差异；28 d后观察组1上述指标明显高于观察组2、3及对照组，观察组2、3与对照组之间比较无显著性差异。其中肌力恢复情况好者炎性因子水平下降明显（P<0.01）。结论:急性格林-巴利综合征作为炎症反应过程，病情越危重，其体内炎性因子越高，持续时间越久，预后也越差。

Abstract:目的：探讨雌二醇对脑梗死/再灌注损伤大鼠海马区血管内皮生长因子（vascular endothelial growth factor，VEGF）表达的影响。方法:雌性成年大鼠，双侧卵巢切除后随机分为3组。雌二醇治疗组给予苯甲酸雌二醇2 mg·kg-1·d -1腹腔注射，连续7 d；模型组、假手术组腹腔注射等量麻油。模型组和雌二醇治疗组分别建立大脑中动脉梗死（MACO）模型，缺血2 h后再灌注48 h。Western 印迹技术和RT-PCR技术分别检测VEGF蛋白及基因表达。结果:大脑梗死/再灌注损伤后VEGF蛋白、基因表达均增加，雌二醇治疗组VEGF表达高于模型组（P<0.05）。结论:雌二醇增加脑梗死/再灌注损伤大鼠海马VEGF的表达。

Abstract:目的：探讨重型脑挫裂伤后诱导型一氧化氮合酶（induced nitric oxide synthase，iNOS）在脑组织中的表达变化及意义。方法：根据损伤后就诊时间将38例重型脑挫裂伤患者分为6组（0～6 h 7例，6～12 h 8例，12～24 h 9例，24～36 h 5例，36～72 h 6例，72 h以上3例），以4例良性脑肿瘤患者手术切除的部分正常脑组织为对照组。尼氏染色法观察重型脑挫裂伤后神经细胞病理变化过程，RT-PCR检测iNOS mRNA的变化，免疫组织化学染色方法观察iNOS阳性细胞的表达变化。结果：尼氏法染色发现损伤区神经细胞减少；RT-PCR结果发现，与正常对照组相比，重型脑挫裂伤后0～6 h创伤区周围iNOS mRNA表达量开始上升，12～24 h达到高峰，随后渐下降，72 h以后仍有表达；免疫组化结果发现iNOS阳性细胞表达增加在伤后12～24 h达到高峰；相关性分析结果显示iNOS mRNA相对灰度值与iNOS阳性细胞数呈高度正相关（r=0.956，P=0.003）。结论：重型脑挫裂伤损伤区及周边皮质iNOS的表达增加，iNOS mRNA的表达与iNOS阳性细胞数呈正相关。

Abstract:目的：比较支气管异物取出术静脉全麻中应用自主呼吸和控制通气的麻醉特点。方法:80例手术随机分为自主呼吸组(S组，40例)和控制通气组(C组，40例)。S组静脉注入芬太尼和异丙酚，患儿术中保持自主呼吸。C组用静脉麻醉药同S组，另静脉加用琥珀胆碱，术中患儿无自主呼吸需手控通气。记录置入硬质支气管镜首次成功率及所用时间、置入支气管镜后呼吸频率、术中患儿体动及缺氧事件和苏醒时间，并行血气分析，术后询问外科医师对手术条件是否满意。结果:术者在C组中更易置入支气管镜，达到100%的首次置入率；术中患儿体动发生率C组低于S组； S组的PaO2\[(117±38) mmHg，1 mmHg=0.133 kPa\]显著低于C组\[(387±56) mmHg\]，而PaCO2显著高于C组\[(53.3±9.5) mmHg vs (38.6±6.4) mmHg,P<0.05\]；S组患儿苏醒时间长于C组(P<0.05)；术者满意率在C组为97.5%，显著高于S组的75%(P<0.05)。结论:在小儿支气管异物取出术中，应用短效肌松剂行控制通气可以为手术提供更为优良的麻醉效果，降低了患儿缺氧风险，术后苏醒更为迅速。

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