Abstract:Objective:To screen for the perioperative risk factors contributing to early mortality in liver transplant recipients,so as to lay a foundation for improving the survival rate after transplantation.Methods: The clinical data of 307 patients,who received liver transplantation (LT) between May 2001 and Sep.2005 in Changzheng Hospital,were retrospectively investigated.Risk factors that might contribute to early postoperative mortality(early mortality was defined as death before the first permission of discharge from hospital) were subjected to univariate analyses.Factors of significance were analyzed by means of Logistic regression to screen for high risk factors.Results: The clinical data of 4 patients were excluded due to incompleteness and the remaining 303 patients were included in the present study.Results of univariate analyses showed that 20 factors were related to early mortality after LT,including the gender,Child-pugh classification,hepatic encephalopathy,hepatorenal syndrome,time of extubation,leukocyte count,preoperative hematoglobin concentration,blood urea nitrogen,creatinine,prothrombin time(PT),serum sodium and potassium level,volume of ascites,duration of operation,urine volume during operation,massive blood transfusion,volume of sodium bicarbonate during operation,PaO2 when breathing air,bilirubin,and model for end-stage liver disease (MELD) score.Results of multifactor Logistic regression analysis showed that 5 factors were independent risk factors of early mortality after LT,including female gender,low serum sodium level,long operative time,large volume of blood transfusion (>7 500 ml),and high MELD score.Conclusion:It is indicated that female gender,low serum sodium level,long operative time,large volume of blood transfusion and high MELD score are risk factors for early motality after LT.

Abstract:Objective:To screen for risk factors for hypoxaemia and heptopulmonary syndrome (HPS) in patients with end-stage liver diseases by analyzing the preoperative incidence of HPS,so as to provide evidence for anaesthesia management during the perioperative peroid.Methods: The clinical data of 214 patients with liver cirrhosis,who received liver transplantation from May 2001 to Sep. 2005 in Changzheng Hospital,were retrospectively investigated.The statistical package of SAS 6.12 was used to analyze the state of oxygenation.The standard of hypoxaemia and HPS was AaPO2>20 mmHg(1 mmHg=0.133 kPa)and PaO2<70 mmHg.The correlation of PaO2 with duration of mechanical ventilation and intensive care unit (ICU) stay was analyzed.Results: It was showed that the 214 patients often suffered from hypoxaemia.PaO2 was found correlated with Child-Pugh classification,hydrothorax,total bilirubin,ammonia,and lactic acid,with the values of r being -0.22,-0.16,-0.17,0.20,and -0.29,respectively.PaCO2 was correlated with haemoglobin and serum albumin,with the values of r being 0.23 and 0.13,repectively.AaPO2was not correlated with spider naevi,liver palms or Child-Pugh class; PaO2 was not correlated with spider naevi or liver palms.PaO2 was negatively correlated with the duration of mechanical ventilation and ICU stay (r=-0.388 81,P=0.000 1;r=-0.176 84,P=0.010 4).Conclusion: Patients with liver cirrhosis often suffer from hypoxaemia; the incidence of hypoxaemia is correlated with preoperative liver function classification and other related factors,and has great influence on duration of technical ventilation and ICU stay.The incidence of hypoxaemia and HPS should be evaluated before liver transplantation in patients with end-stage liver cirrhosis,so as to benefit perioperative management.

Abstract:Objective:To investigate the protective effect of astilbin on warm ischemia/reperfusion-induced liver injury and to study the related mechanism.Methods:C57BL/6 mice were randomly divided into four groups (n=8): sham-operated group (Sham),model control group (I/R),low dose astilbin treatment group (10 mg/kg) and high dose astilbin (40 mg/kg) treatment group. Mice in the two treatment groups were intraperitoneally injected with astilbin 24 hours and one hour before ischemia. Then 70% hepatic ischemia/reperfusion model (the left and middle hepatic lobe) was established. Mice in the I/R model control group and the sham operation group were administered with the same volume of normal saline. The blood sample and liver tissue samples were collected 90 min after ischemia and 6 h after reperfusion. Serum ALT activity was detected as an indicator of liver function damage and the content of MPO in liver tissues were detected by ELISA. The pathological changes of the liver were observed. The expression of IL-10 in liver tissues was detected by Western blotting and the expression of IL-10 mRNA was detected by semi-quantitative RT-PCR.Results:Astilbin treatment can effectively lower the serum ALT level and MPO level in the liver tissues in some I/R mice. It could also improve the pathological manifestations of the liver. Compared with the I/R model control group,IL-10 protein levels gradually increased in the two treatment groups,which was consistent with the result of RT-PCR (low dose group P<0.05; high dose group,P<0.01).Conclusion:Astilbin can effectively reduce the inflammatory response after liver warm ischemia-reperfusion induced injury,effectively improve the mouse liver function and pathological damage,which might be related to the upregulation of IL-10 expression in the liver tissues.

Abstract:Objective:To investigate the protective effect of sinomenine on the cold ischemia/reperfusion injury during orthotopic liver transplantation (OLT) in rats.Methods: OLT were performed in normal rats using Kamada’s two-cuff technique.The rats were randomly divided into sham operation,control and two sinomenine groups.Animals in sinomenine groups were treated with low (40 mg/kg) or high dose (80 mg/kg) of sinomenine.The serum and tissue samples of rats in each group were collected 2,6,12, and 24 h after reperfusion,and the one-week survival rate was observed.The apoptosis index (AI) of liver cells after OLT was detected by TUNEL.The expression of TNF-α and IL-1β mRNA in the liver were detected by RT-PCR.Results: Compared with the control group,the level of ALT was significantly decreased in the two sinomenine treatment groups at different time points after transplantation,and their one-week survival rates were also significantly increased (75%,75% vs 12.5%,P<0.01).Compared with the control group,the AI of liver cells was markedly decreased in sinomenine treatment groups (P<0.01).Sinomenine treatment also significantly decreased the expression of TNF-α and IL-1β mRNA (P<0.01) and greatly ameliorated the focal necrosis of hepatocytes and sinusoidal endothelial cells (SECs).Conclusion: Sinomenine can inhibit hepatocyte apoptosis by inhibiting the synthesis of TNF-α and IL-1β.sinomenine can prevent hepatic cells and SECs from cold ischemia/reperfusion injury during orthotopic liver transplantation in rats.

Abstract:Objective:To investigate the protective effect of repeated ischemic preconditioning (IP) in a short period on rat liver graft from ischemic reperfusion injury (IRI).Methods: Male Sprague-Dawley rats were used as donors and recipients for orthotopic liver transplantation.The period of cold preservation and anhepatic phase were 120 min and 14 min,respectively.Fifty-four rats were randomly divided into 5 groups.Control group (group A,n=6) received no operation of liver transplantation; liver transplantation group (group B,n=12) received orthotopic liver transplantation by the cuff technique with donor livers perserved for 2 h in 4℃ Ringer’s lactate solution before transplantation; and ischemic precondition group included three subgroups:group C1,C2,and C3(n=12).For group C,the grafts were harvested and then perserved for 2 h in 4℃ Ringer’s lactate solution after ischemic preconditioning,and then used for orthotopic liver transplantation.An IP cycle included 5 min interruption of the portal veins and hepatic arteries of liver,and reflow for 5 min.The C1,C2,and C3 animals were subjected to one,two and three circles,respectively.The activities of AST,ALT and LDH in the sera were examined and the capacity of bile per min was examined 6 hours after operation.The expression of Bcl-2 protein in hepatic tissue was determined with immunohistochemistry.Moreover,the hepatocellular apoptosis were detected by TdT-mediated dUTP-biotin nick end labeling (TUNEL) and the changes of liver ultrastructure were observed under electron microscope.Results: The activities of ALT,AST,and LDH in group B were significantly higher and the capacity of bile per min was significantly lower than those in group A(P<0.01).The activities of ALT,AST,and LDH in group C were significantly higher than those of group A (P<0.01),but were significantly lower than those of group B (P<0.05),and decreased with the increase of IP cycle.Cell morphology examination showed that the changes of graft structure was slighter in group C than in group B,and the expression of Bcl-2 was increased and the apoptotic index was lowered (P<0.01 or P<0.05).Conclusion: Ischemic precondition can relieve IR-induced injury of the donor liver.The protective effect is more potent with repeated IP cycles in a short time.

Abstract:Objective:To observe the effect intraperitoneal injection of low dose ketamine on thermal hyperalgesia and expression of P2X4 receptor in spinal dorsal horn of rats with chronic constriction injury (CCI) of sciatic nerve,and to explore the potential role of P2X4 receptor in the neuropathic pain.Methods: Totally 24 Sprague-Dawley rats were randomly divided into 3 groups (n=8): group S (sham group),group C: CCI + normal saline; and group K: CCI+ketamine (10 mg·kg-1）.Rat CCI model was used in the latter 2 groups.Three days after operation the thermal withdrawal latency (TWL) was determined to confirm the thermal hyperalgesia.Rats in group K were given low dose of ketamine (10 mg·kg-1）and those in group C were given the same volume of normal saline for 7 days after operation.Animals in group S only had sciatic nerve exposed,with no ligation or drugs.TWL was determined 1 day before and 1,3,7 days after the operation.The expression of P2X4 receptor was assessed 7 days after the operation using immunohistochemistry.Results: The TWL values were similar between the 3 groups before operation.The value in group S was slightly decreased after operation compared with before operation.Compared with the pre-operation,group S,and group C,the TWL value of group K began to gradually increase 3 days after operation till day 7 after operation(P<0.05).On day 7 after operation,the TWL value was significantly higher than group C (P<0.05),but was still lower than that in group S (P<0.05).Immunohistochemistry showed that the expression of P2X4 receptor in group C,K were significantly higher than that of group S (P<0.01),and the expression in group K was significantly lower than that in group C(P<0.05).Conclusion: Intraperitoneal injection of ketamine can partly relieve the thermal hyperalgesia in rats with CCI of sciatic nerve,which might be related to the inhibition of P2X4 receptor expression in the spinal dorsal horn.

Abstract:Objective:To establish a lung metastasis mouse model of clear cell renal cell carcinoma (CCRCC) and screen for lung metastasis CCRCC cell line,so as to lay a foundation for future study.Methods: RCC05-TXJ,a CCRCC cell line established by our lab,was used in the present study.The cells were proliferated and the cell suspension was subcutaneously injected into the NOD-SCID mice.The newly-formed tumors were transplanted into the renal capsule of NOD-SCID mice (in situ),and the formed tumors were harvested and again transplanted into NOD-SCID mice.A stable mouse model with CCRCC lung metastasis was established following several cycles of the transplantation.This transplantation cycle was repeated until the development of stable lung metastasis.The lung metastasis tissues were then injected into renal capsule for several cycles to induce the lung metastasis.The metastasis tissues were also primary cultured and proliferated and the cell suspensions were injected into renal capsule until the development of lung metastasis.The sub-cell lines which could stably produce lung metastasis were screened.During the screening,the in situ tumor information,lymphatic and pulmonary metastasis were all recorded in detail.Immunohistochemistry analyses were used to examine the expression of CA9 (a marker of renal cells) and CD133 (a marker of stem cells) on the metastatic CCRCC cells.Results: The lung metastasis was observed in the second cycle of the in situ transplantation of RCC05-TXJ in NOD-SCID mice.When the metastatic lung carcinoma tissues were used for in situ transplantation,lung metastasis was successfully established during the 3-6 cycles.CCRCC cells were successfully cultured with lung metastatic tissues,and the cell suspension of the passage 6 cells caused lung metastasis after implanted in the renal capsule.The result was stable in two repeated cycles.The lung metastatic RCC05-TXJ-L cells grew fast (average passage time was 2 days),and had a short in situ tumor-forming time (1 week).They also had a strong potential for lung metastasis and high expression of CA9 and CD133.Conclusion: Repeated in situ transplantation of RCC05-TXJ can be used to establish stable lung metastasis mice model of CCRCC.The selected RCC05-TXJ-L sub-cell line can stably induce lung metastasis in NOD-SCID mice.

Abstract:Objective:To study the time- and concentration-dependent effect of DNA methyltransferase inhibitor 5-aza-2′-deoxycytidine(5-aza-dC) and histone deacetylase inhibitor trichostatin A(TSA) on the expression of H19 gene in nuclear donor cells.Methods: Donor cells (fibroblast-like cells from tail tip of mice) were treated with 5-aza-dC and TSA at different concentrations for different time periods.The expression of H19 was determined by real-time PCR after treatment.Results: The expression of H19 was not significantly different between groups treated with different concentrations of 5-aza-dC,but the expression of H19 was increased with the increase of treatment time.TSA decreased H19 expression,but not in a time-dependent manner.Conclusion: Long time exposure to low concentration of 5-aza-dC can increase the expression of H19; TSA can decrease the expression of H19,which provides a basis for selection of epigenetic modification inhibitor and later study on imprinting in somatic cell nuclear transfer.

Abstract:Objective:To construct a retroviral vector carrying both enhanced green fluorescent protein(EGFP) and human insulin gene for transfecting the bone marrow mesenchymal stem cells (BMSCs),and to observe the treatment effect of transfected BMSCs after transplanted into diabetes mice.Methods:Two gene segments of insulin-IRES-EGFP and IRES-EGFP were obtained by overlap extension PCR technology,and then the 2 segments were cloned into retroviral vector (pMSCV).The vectors were used to transfect BMSCs after packaged with PT67 cells.The expression of EGFP was observed by inverted fluorescence microscope and the expression of insulin gene was examined by RT-PCR in the infected BMSCs.The transfected BMSCs with 2 viruses were transplanted into diabetic mice separately.The blood glucose levels and body weights of mice were examined in 2 groups,and the results were compared with those of normal control group.Results:The recombinant retroviral vectors pMSCV-insulin-IRES-EGFP and pMSCV-IRES-EGFP were successfully constructed.The BMSCs infected by both vector stably gave out green fluorescence under microscope; the cells infected with pMSCV-insulin-IRES-EGFP also stably expressed human insulin gene.The blood glucose level of the mice transplanted with BMSCs transfected with pMSCV-insulin-IRES-EGFP was significantly lower than that transplanted with BMSCs transfected with pMSCV-IRES-EFGP (P<0.05),but was still higher than that of the control group(P<0.05).Thirty-five days after transplantation,the body weight of the pMSCV-insulin-IRES-EGFP was significantly higher than that of the pMSCV-IRES-EGFP group(P<0.05).Conclusion: The insulin-EGFP recombinant vector can be traced easily,and the expression of EGFP does not affect the expression of insulin.Transplanting the BMSCs transfected with insulin fusion gene can partially relieve the symptoms of the diabetes.

Abstract:Objective:To construct recombinant human JNK3 adenovirus and study its influence on the proliferation of human neuroblatoma SH-SY5Y cells, and to study its influence on epirubicin-induced apoptosis.Methods: The full-length JNK3 cDNA fragment was amplified by PCR from the pDBLeu-JNK3 plasmid to construct the shuttle plasmid AdTrack-CMV-JNK3.Then the linearized shuttle plasmid was co-transformed into BJ5183 bacteria with backbone vector pAdEasy-1 to obtain the recombinant adenoviral plasmid Ad-JNK3 by homologous recombination.The linearized recombined adenovirus Ad-JNK3 was then transfected into HEK293 cells for packing and amplification.Viral titers were measured by endpoint dilution assay.Ad-JNK3 was identified by PCR.The expression of Ad-JNK3 in SH-SY5Y cells was detected by Western blotting assay.The influence of Ad-JNK3 on the proliferation of SH-SY5Y cells was assayed by MTT after cells were infected by 100 pfu/ml adenovirus.The cell apoptosis induced by 0.5 μg/ml epirubicin was detected by flow cytometry method and agarose gel electrophoresis.Results: The recombinant adenoviral shutter vector AdTrack-CMV-JNK3 and recombinant adenoviral vector Ad-JNK3 were successfully constructed as identified by sequence analysis.PCR assay showed that adenovirus Ad-JNK3 contained JNK3 gene.After amplification in packing cell HEK293,6.5×1010 pfu/ml titer of Ad-JNK3 was obtained.Western blotting assay showed that JNK3 protein was expressed in SH-SY5Y cells.After infected by 100 pfu/ml adenovirus,the proliferation of SH-SY5Y cells was inhibited by 28.08% with MTT method.Flow cytometry showed that Ad-JNK3 significantly promoted the apoptosis of SH-SY5Y cells induced by epirubicin.The DNA ladder of agarose gel electrophoresis was clearly seen.Conclusion: Ad-JNK3 can obviously inhibit the growth of SH-SY5Y cells and promote apoptosis induced by epirubicin,which provides a solid foundation for further studies on the function of the JNK3 gene and applying it in gene therapy for human neuroblastoma.

Abstract:Objective:To establish a method for isolating and culturing non-small cell lung cancer-derived vascular endothelial cells (NSCLCDVECs) and to observe their characteristics in vitro.Methods: Enzyme digestion method was used to isolate microvascular fragments from human non-small cell lung cancer tissues,and explant culture method was used for primary culture of the microvascular endothelial cells,then the cells were purified by local digestion and differential adhesion method.The obtained NSCLCDECs were identified by optical microscope,immunocytochemistry and immunofluorescence methods.Results: The NSCLCDECs were positive for FⅧ-RAg and CD34.Microscopy showed that the cells grew well and could be passaged.Conclusion: We have established a method for isolating and culturing NSCLCKVECs,and the resultant cells possess a great value for research on anti-vascularizaton and anti-tumor agents.

Abstract:

Abstract:Objective:To design and synthesize novel norcantharidin derivatives and to investigate their anti-tumor activities in vitro.Methods: Novel norcantharidin analogues were synthesized through several steps using furan and maleic anhydride as the starting material.All the target compounds were confirmed by 1HNMR,ESI-MS and element analysis,and screened against 4T1(breast cancer cell).The compound 7a and 7c with better activity were further screened against LLC(lung cancer cell),SK-HEP-1(human hepatoma cell) and ABAE(vascular endothelial cell).Results: Totally 18 novel target compounds were obtained.The results of anti-cancer test showed that compounds 7a and 7c had anti-tumor activity and less damage to normal cells.Conclusion: Compounds 7a and 7c have inhibitory effect against 4T1 cells,LLC cells and SK-HEP-1 cells.Compound 7c shows less damage to ABAE(normal vascular endothelial cells) than norcantharidin and is worth further studying.

Abstract:Objective:To observe the biological behaviors of colorectal cancer LS174 cells before and after pcDNA3.0-hugl-1 transfection,so as to investigate the association of hugl-1 with colorectal cancer.Methods: The eukaryotic expression vector pcDNA3.0-hugl-1 was constructed and transfected into LS174 cells.RT-PCR and Western blotting methods were used to analyze the expression of hugl-1 mRNA and protein in LS174 cells before and after transfection.Soft agar colony formation assay,wound-healing experiment,adhesion assay and Matrigel invasion assays were used to study the effects of hugl-1 expression on the proliferation,adhesion，movement and invasion in LS174 cells.Results: The recombinant plasmid pcDNA3.0-hugl-1 was successfully constructed.RT-PCR and Western blotting showed that the hugl-1 expression was higher in cells transfected with pcDNA3.0-hugl-1 than in those un-transfected or empty vector-transfected cells (P＜0.05).The colony forming rates showed no significant difference between the 3 groups (35.76%,33.91% and 32.23%). Wound healing assays showed a significant reduction in the migrated cells in pcDNA3.0-hugl-1 transfected cell clones(82.14±7.62) compared with un-transfected cell lines(142.37±6.12)and empty vector transfected cell line (135.61±3.74,P＜0.05). Attachment assays revealed enhanced adhesion of the pcDNA3.0-hugl-1-transfected cell clones compared with the other 2 groups.Matrigel invasion assay showed a significant reduction in the invasive ability of the pcDNA3.0-hugl-1-transfected cell (63.7±8.0) in comparison to the un-transfected cells(156.3±13.0)and pcDNA(-) cells(158.3±16.5,P＜0.05).Conclusion: Overexpression of hugl-1 eukaryotic expression vector can decrease migratory and invasive ability of LS174 cells,but has no influence on cell proliferation.

Abstract:Objective:To investigate the chemical constituents in the BuOH extract of Asterias amurensis.Methods: The compounds were obtained by a combination of extraction and purification with suitable solvent and multiple column chromatographies; their structures were determined based on spectral analysis and physicochemical properties.The bioactivities of 5 compounds were tested.Results: Five compounds were isolated and identified,and they were amurensoside D(Ⅰ),tryptophan Ⅱ,and 3 sulfated sterols,including 3β-O-sulfated-6α,20β-diol-cholest-9(11)-en-23-one sodium salt(Ⅲ),3β-O-sulfated-6α-ol-cholesta-9(11),20(22)-dien-23-one sodium salt(Ⅳ),and 3β-O-sulfated-6α-ol-ergost-9(11)-en-23-one sodium salt(Ⅴ).Conclusion: TryptophanⅡ is obtained from this species for the first time; compoud Ⅲ and compound Ⅴ exhibit potent inhibitory activity against Pyricularia oryzae P-2b.

Abstract:Objective:To evaluate the relationship of the shape,location of superficial calcification with surface ulcer,intraplaque hemorrhage in the atherosclerotic carotid plaque (ACP).Methods:Fifty-one patients received pre- and post-contrast high-resolution 3T MRI for ACP and a total 183 layers with superficial calcification were enrolled in our study.The shape of the superficial calcification was categorized as irregular type(dotted/arcuated) and patch type; the location of the calcification was categorized as marginal type and central type.Intraplaque hemorrhage and surface ulcer at the same or adjacent slice to the superficial calcification were identified.Chi-Square test was used to analyze the relationship of the shape,location of superficial calcification with surface ulcer and intraplaque hemorrhage.Results::We found that 122(66.7%) of the 183 slices had intraplaque hemorrhage and 86(47%)slices had surface ulcer.We also found that 158 slices had irregular type(dotted/arcuated)calcification and 142 slices had marginal calcification.Irregular superficial calcification was found more frequently with intraplaque hemorrhage than patch type (72.8% vs 28%,P<0.01).Marginal type had higher possibility of intraplaque hemorrhage than central type did (71.1% vs 51.2%，P<0.05).However,the incidence of surface ulcer was not significantly different between different types and locations of superficial calcification.Conclusion:It can be concluded that superficial calcification is an important factor for instability of atherosclerotic carotid plaque.The shape and location of superficial calcification are important factors of intraplaque hemorrhage.

Abstract:Objective:To examine the relationship between brain natriuretic peptide (BNP) and the recurrence of atrial fibrillation (AF) after successful electrical cardioversion (EC) by a meta analysis approach.Methods: Based on PubMed,Medline and China National Knowledge Infrastructure (CNKI) databases,related literatures,published from 1980-2008,were searched; a manual search was also performed.The included trials were analyzed by RevMan4.2 software for meta analysis.Results: Totally seven studies were finally included.The analysis revealed that the baseline BNP level were greater in patients with AF recurrence than those without AF recurrence,with the standardized mean difference in the BNP levels being 0.82 units (95% confidence interval 0.12 to 1.52).The test for overall effect Z-score was 2.30,with a P value of 0.02; there was significant difference.Conclusion: It is suggested that the higher BNP level after successful electrical cardioversion is associated with greater risk of AF recurrence.The level of BNP may serve as an indicator for AF recurrence after successful EC.

Abstract:Objective:To develop a system to screen for the effective siRNA sequence targeting HBs gene and to identify the interference efficiency.Methods:Three HBs-targeting siRNA segments (HBs-siRNA1,HBs-siRNA2,and HBs-siRNA3) were designed,synthesized and cloned into pSUPER vector to construct three recombinant plasmids pSUPER-HBs-siRNA,which were then transfected into human embryonic kidney 293T cells together with HBV plasmid.The transfection efficiency was observed 72 h later,the interference efficacies of the 3 segments were identified by real-time PCR and ELISA analysis,and the best one was identified.Results:Three recombinant plasmids of pSUPER-HBs-siRNA were constructed successfully and effectively transfected into 293T cells to induce RNAi,with a transfection rate higher than 70%.The results of real-time PCR and ELISA analysis showed that HBs-siRNA2 silenced the HBs gene expression by more than 80%.Compared with HBs-siRNA2,HBs-siRNA1 and HBs-siRNA3 did not demonstrate obvious interfering effect (P<0.05).Conclusion:We have successfully constructed 3 siRNA sequences targeting HBs,and pSUPER-HBs-siRNA2 can effectively silence HBs genes,which paves a way for future study.

Abstract:Objective:To observe the effect of glucocorticoid exposure of pregnant rats on hippocampus SGK expression in rat offsprings,and to explore the related mechanism.Methods:Pregnancy was determined by examining vaginal smear in the morning of cohabiting.Pregnant rats were divided into control group and dexamethasone group,with each group containing 10 rats.Dexamethasone group was subcutaneously injected with dexamethasone(0.1 mg/ \[kg·d\] ) and control group was injected with normal saline.The hippocampuses of rat offsprings were collected 1 d，7 d,and 42 d after birth.Real-time PCR was used to determine SGK and GR mRNA expression.Results:The body weight of rat offsprings in the dexamethasone group was obviously lower than that in the control group at birth,then those in the dexamethasone group had a rapid growth.Prenatal exposure to dexamethasone significantly decreased the expression of SGK and GR mRNA in the hippocampuses of rat offsprings (1 d,7 d and 42 d after birth) compared with control group (P<0.05).Conclusion:Prenatal exposure to glucocorticoid in pregnancy stage can decrease the hippocampus expression of SGK and GR in rat offsprings.

Abstract:Objective:To transfect rabbit conjunctival epithelial cells in vitro with recombinant adeno-associated virus 2-mediated enhanced green fluorescent protein (rAAV2-EGFP),so as to lay a foundation for studying the proliferation and differentiation of conjunctival epithelial stem cells.Methods: Rabbit conjunctival epithelial cells were cultured in vitro.The second passage of rabbit conjunctival epithelial cells were transfected with rAAV2-EGFP in various multiplicity of infection (MOI=104,105,106).The expression of green fluorescent protein was examined 1,3,5,and 7 days after transfection and the transfection rate was calculated.MTT assay was used to study the influence of rAAV2-EGFP on the growth of cells.Results: With the increase of MOI value and the prolongation of transfection,the expression of EGFP was gradually increased and reached its peak on 7-8 days after transfection.MTT assay showed that there was no significant difference between the MOI group and the control group.Conclusion: rAAV2-EGFP can efficiently transfect rabbit conjunctival epithelial cells and have no influence on the proliferation of the cells.

Abstract:Ultrasmall superparamagnetic iron oxide (USPIO),as the contrast agent of MRI,possesses two major properties:long half time in the plasma and specific binding with macrophages.Compared with gadolinium,widely-used in clinic presently,USPIO has its unique advantages in diagnosis of central nervous system diseases,though which still need further clinical verification.This article reviews the application of USPIO in MRI diagnosis of some central nervous system diseases.

Abstract:Objective:To determine the contents of eight ginsenosides in the root of Panax ginseng by high performance liquid chromatography (HPLC). Methods:Panax ginseng was extracted with methanol solution for 30 min.The HPLC condition was as follows-column:Agilent SB-C18(3.0 mm×100 mm,3.5 μm); mobile phase:A was ACN,B was H2O,with gradient elution;the gradient of A phase:18%-20%(0-14 min),20%-29%(14-20 min),29%(20-25 min),29%-37%(25-34 min),37%-55%(34-40 min); flow speed:0.6 ml/min; detection:203 nm; temperature of column:25℃;injection volume:10 μl. Results:Eight ginsenosides (Rb1,Rb2,Rb3,Rc,Rd,Re,Rf and Rg1) were separated at baseline within 35 min with good linearity (r=0.9999).The result of intra-day and inter-day precisions,limits of detection and quantitation were all within the normal range. The recovery rates(n=3) were 101.07%,98.72%,101.57%,101.71%,102.12%,99.58%,98.62%,and 100.12%,respectively. The contents of eight ginsenosides in 6 different batches of Panax ginseng from 3 sources were determined.Conclusion:The present method is rapid,simple,accurate and can be used to control the quality of Panax ginseng.

Abstract:目的:分析第二军医大学长征医院社区获得性肺炎(CAP)的病原学分布及常见细菌的耐药性,为CAP的经验治疗提供依据。方法:收集2003年9月至2006年12月间我院收治的247例社区获得性肺炎患者,采集痰、支气管吸出物和血标本,进行细菌学和非典型病原体的检测。结果:247例患者中,共有128例患者病原学检测阳性,混合感染20例(15.6％)。居前3位的病原体分别为肺炎支原体39例(26.4％),流感嗜血杆菌23例(15.5％),肺炎链球菌17例(11.5％),肺炎衣原体17例(11.5％)。同时应用PCR和血清学两种方法对标本进行非典型病原体的检测,肺炎支原体、肺炎衣原体和嗜肺军团菌的诊断符合率分别为84.6%(33/39)、23.5％(4/17)和14.3％(1/7)。就诊前有无使用抗菌药物对于苛养菌检出率的影响显著。结论:流感嗜血杆菌和肺炎链球菌仍是CAP最常见的致病细菌,但非典型病原体尤其是肺炎支原体感染在CAP中已占据重要地位。应用血清学检测方法联合PCR方法或尿抗原检测,可以增加非典型病原体检测的敏感性和特异性。

Abstract:目的:探讨熵指数在老年患者七氟醚吸入或丙泊酚靶控输注诱导时的应用价值。方法:拟行气管插管全麻的老年患者40例,ASA Ⅰ~Ⅱ级,年龄≥65岁,随机分为七氟醚组(S组)和丙泊酚组(P组),分别以七氟醚吸入和丙泊酚靶控输注诱导,诱导期间调节熵指数维持在40~50,记录诱导、插管前后的血流动力学变化。结果:两组患者各项参数在诱导后均下降,气管插管后又上升,P组平均动脉压(MAP)在插管前明显低于S组(P<0.01),两组各时点的RE及SE数值无统计学差异,两组患者气管插管前后血流动力学增加值ΔHR、ΔMAP与熵指数增加值ΔSE、ΔRE之间没有相关性。结论:对于老年患者,熵指数可以反映七氟醚或丙泊酚靶控输注麻醉诱导时的麻醉深度,但不能准确预测气管插管时的心血管反应。

Abstract:目的:观察外源性血管紧张素(Ang)Ⅱ灌注大鼠气道对肺上皮屏障的损伤作用,并探讨其可能的损伤机制。方法:Wistar大鼠随机分为5组:PBS对照组、Ang 6、20 h组、Ang+zVAD-fmk组、Ang+氯沙坦组(n=10),各组大鼠分别经气管灌注PBS、Ang(10 μmol/L)磷酸盐缓冲溶液、Ang(10 μmol/L)+zVAD-fmk(60 μmol/L)、Ang+氯沙坦(losartan,100 μmol/L),另设10只大鼠不作任何处理作为空白对照组。大鼠经气管灌入相应溶液后在体灌洗动物肺脏,测定灌洗液中的血红蛋白(Hb)和灌洗肺前由静脉注入的荧光物质(BODIPY)标记的白蛋白含量,判断肺损伤；取肺组织标本进行病理学观察；采用末端脱氧核苷酸转移酶标定法(TUNEL)和抗caspase 3免疫标定方法检测肺泡上皮细胞凋亡。结果:灌注Ang 6 h后,支气管和肺泡上皮细胞内标定的断裂DNA片段(P<0.05)和活性caspase 3(P<0.01)显著增加,但单纯灌注磷酸盐缓冲溶液没有作用；同时灌注caspase抑制剂zVAD-fmk或Ang受体1阻断剂氯沙坦均阻断Ang对DNA 和caspase 3的作用。灌注Ang 6 h后,肺泡灌洗液中荧光标定的白蛋白含量(P<0.01)和Hb含量(P<0.05)亦显著增加；zVAD-fmk或氯沙坦亦阻断支气管肺泡灌洗液中荧光白蛋白含量和Hb含量的变化。结论:在体大鼠呼吸道内灌注外源性Ang可介导肺泡上皮细胞凋亡；Ang诱导的在体肺泡上皮细胞凋亡可能与Ang受体1亚型介导的激活caspase 3信号转导通路有关。

Abstract:目的:比较经尿道双极等离子前列腺切除术（bipolar plasmakinetic resection of prostate,PKRP）与传统经尿道前列腺电切术（transurethral resection of the prostate,TURP）治疗良性前列腺增生（BPH）的疗效，评价PKRP的安全性和有效性。方法:回顾性分析2003年12月至2008年6月我院收治的156例采用PKRP治疗的BPH患者临床资料（PKRP组），观察手术前后最大尿流率（Qmax）和IPSS(international prostate symptom score)评分的变化，并与同期采用传统TURP术式的140例BPH患者（TURP组）进行比较。两组患者年龄、病程均无统计学差异，具有可比性。结果:两组患者手术过程顺利，均未发生术中转开放手术的事件。PKRP组患者手术时间、切除前列腺组织的质量与TURP组无统计学差异；PKRP组术中出血量明显少于TURP组，PKRP组前列腺包膜切穿率、TURP综合征发生率、尿失禁发生率均明显低于TURP组（均P<0.05）。术前两组患者Qmax、IPSS评分无统计学差异；术后3个月，两组患者Qmax均明显高于术前，IPSS评分均明显降低（P<0.05），而两组间各指标无统计学差异。结论:PKRP术治疗BPH与TURP疗效相当，能部分解决TURP术出血量多、TURP综合征发生率高的缺陷，值得在基层医院推广使用。

Abstract:目的:探讨不明来源的双侧颈部淋巴结转移性鳞癌的治疗策略。方法:回顾性分析1990年～2003年我院收治的50例双侧发病原发灶不明的颈部淋巴结转移性鳞癌患者的临床资料,比较不同治疗手段的效果。50例患者男性44例,女性6例,年龄29～73岁,中位年龄52.5岁；临床分期：N2c占64% (32/50),N3占36% (18/50)。30例患者行双颈淋巴结清扫术。50例患者均行放疗,剂量为50～66 Gy,全颈部放疗22例,行颈面部和(或)颈胸部联合放疗28例,治疗后总体的5年生存率为36%(18/50)。结果:30例患者行双侧颈淋巴结清扫术后配合放疗5年生存率及局部肿瘤复发控制率分别为46.7%及86.7%,明显高于单纯放疗患者20%及45%(P<0.05)。行颈面和(或)胸部联合放疗的患者5年生存率及原发灶肿瘤控制率均高于行全颈部放疗的患者(42.8%、92.9% vs 27.3%、68.2%,P<0.05)。结论:不明原发灶的双侧颈部淋巴结转移性鳞癌手术联合术后放疗能提高患者生存率,若无法手术应扩大放疗照射范围以提高生存率。

Abstract:目的:建立测定人血浆中替米沙坦浓度的LC-MS/MS方法,并用该法研究替米沙坦片在健康人体内的药代动力学。 方法:采用LC-MS/MS测定人血浆中不同时间点替米沙坦的浓度,以地西泮为内标,血浆样品用乙腈沉淀蛋白,色谱柱为Agilent Zorbax SB-C18 (100 mm×2.1 mm,3.5 μm),流动相为乙腈-0.3%甲酸水溶液(55∶45),流速0.3 ml/min,柱温30℃。 结果:替米沙坦的线性范围为2.5～500 ng/ml (r=0.995 7),最低定量限为2.5 ng/ml(S/N>10),日内和日间RSD<8%。 结论:本法操作简单,选择性好,灵敏度高,适用于替米沙坦的人体药代动力学研究。

Abstract:[Abstract] Objective To investigate the influence of different modes of blood purification treatment on atherosclerosis (AS) in patients with maintained hemodialysis(MHD) by measuring the serum levels of advanced oxidation protein products (AOPP) and the datas of carotid artery Methods 72 patients with MHD were assigned to receive the treatment of different modes of blood purification : ①hemodialysis（HD）, ②hemodiafiltration（HD+HDF）, ③ hemodialysis combined by continuous renal replacement therapy（HD+CRRT）.Serum AOPP、interleukin-6(IL-6)、monocyte chemotactic protein-1(MCP-1)、high sensitivity c-reactive protein (hs-CRP) , IMT and mass of carotid artery plaque were detected before and after one year treatment. Results The levels of serum AOPP、IL-6、MCP-1、hs-CRP in MHD patients were siginificantly higher than those in healthy controls (p<0.001). The levels of serum AOPP decreased after the treatment of HDF and CRRT (p<0.001) . After 12 –month treating , the level of serum AOPP increased of HD( P<0.01), meanwhile IMT and the mass of carotid artery plaques in patients with HDF and HD+CRRT were lower than that inpatients with HD. Serum level of AOPP was strongly associated with IMT and mass of carotid artery plaque (r=0.79, r=0.67) . Conclusions AOPP may be one of the risk factor for atherosclerosis. HDF and CRRT can clear AOPP effectively, and can improve the survival rate of MHD patients.

Abstract:目的:应用加权TOPSIS法对医院医疗工作质量作出客观评价,为医院管理部门制定决策提供依据。方法::应用加权TOPSIS法从医疗工作量、医疗质量和医疗效率3个方面对我院1998~2007年工作质量进行综合评价和分析。结果:评价结果与医院的实际情况相符,2007年最佳,1998年最差。结论:TOPSIS法具有计算简便、结果合理、应用灵活的特点,适宜在不同级别、不同层次的医院推广使用。

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