Objective To establish a reliable approach for quantification of colony forming unit(CFU)of Mycobac-terium tuberculosis (M.tb)by measuring optical density(OD).Methods M.tb suspension H37Ra was prepared using low-power ultrasonic or glass bead beating methods,and was two-fold serially diluted,OD at 600nm (OD600)of each dilution ratio was measured respectively,OD600 and dilution curve were analyzed to determine the optimum approach for preparing bacterial suspension,linear range of OD600,as well as linear relationship between OD600 and CFU.Results OD600 was 0.1 -0.6,linear regression analysis of OD600 and dilution ratio within linear range revealed that correlation coefficient (R2 )of glass bead beating and low-power ultrasonic methods were 0.98 and 1 .00 respectively,both presented a good correlation,low-power ultrasonic method was better than glass bead beat-ing method,bacterial suspension dispersed more evenly.Linear regression analysis results of OD600 and CFU val-ues showed that the regression equation of glass bead beating method and low-power ultrasonic method were CFU=2.35×107 ×OD600+4.42×105 and CFU=3.26×107 ×OD600+6.89×105 respectively.Conclusion Low-power ultrasonic method is a good method for preparation of M.tb suspension,combined the measurement of OD600 value, it can be a reliable and rapid method for quantitative analysis of M.tb.

Objective To investigate the protective effects and the mechanisms of 17β-estradiol on the propofol-induced neuroapoptosis in primary cultured rat cortical neurons.Methods The neurons were cultured for 7 days and then divided into three groups: vehicle-control group ( treated with equal volume of 20% intralipid ) , propofol-treated group ( treated with 500μmol/L propofol) , and propofol plus 17β-estradiol treated group ( treated with 500μmol/L propofol and 0.1 μmol/L 17β-estradiol).12 hours after the treatment, neuroapoptosis was detected by Hoechst 33258 staining and TUNEL assay, and the levels of Bcl-2, Bax and cleaved caspase-3 proteins were detected by Western blot.Results Compared with the vehicle-control group, the neuroapoptosis increased greatly ( P<0.01 ) , Bcl-2 level reduced ( P <0.01), Bax and cleaved caspase-3 levels increased greatly (P<0.01), and Bcl-2/Bax ratio reduced significantly (P<0.01).Compared with the propofol-treatment group, the neuroapoptosis decreased greatly ( P <0.01), Bcl-2 level increased ( P<0.01 ) , Bax and cleaved caspase-3 levels reduced greatly ( P <0.01 ) , and Bcl-2/Bax ratio increased greatly ( P <0.01 ) . Conclusions 17β-estradiol can protect cortical neurons against propofol-induced cortical neuroapoptosis by regulating the expression of Bcl-2 and Bax.

ObjectiveTo investigate the effects of selective splenectomy on modulation of portal vein flow and prevention of small-for-size syndrome (SFSS) in living donor liver transplantation.MethodsTwenty six recipients who received LDLT from September 2007 to March 2008 were reviewed.The data of the portal vein flow of these recipients were collected during the operation.Simultaneous splenectomy was performed in patients with portal blood flow ＞250 ml/(min · 100g).No splenectomy was performed when the portal blood flow was less than 250 ml/(min · 100g).The effect of selective splenectomy on modulation of portal vein flow and whether splenectomy prevented the occurrence of SFSS were analyzed.ResultsThe portal vein flow decreased significantly after splenectomy in 8 patients who received splenectomy (P＜0.01),No SFSS occurred in the patients with or without splenectomy.Actual graft-to-recipient weight ratio (GRWR) of patients with splenectomy was significantly smaller than those with no splenectomy (P=0.044).The portal vein flow of patients with splenectomy was much higher than those with no splenectomy (P＜0.01).ConclusionAccording to the portal blood flow,selective splenectomy in LDLT decreased the portal vein flow and prevented the incidence of SFSS.

Aim To investigate the protective effect of naringenin on isoniazid and rifampicin induced hepatotoxicity and the role of CYP 3A4.Methods Isoniazid and rifampicin were added to culture media for QSG-7701 cells and cultured for 48 hours. Narringenin, 1,5 and 25 mg·L~(-1) in final concentration,was added concomitant with isoniazid and rifampicin. The culture media and cells were collected and the activities of lactate dehydrogenase were detected with chromatometry. The ratio of extra/intracellular lactate dehydrogenase was calculated as the release rate of lactate dehydrogenase. Cells were incubated with midazolam for 2 hours after treatment with durgs and the concentration of midazolam in the incubation media was determined with HPLC-MS.Results Compared with control group, isoniazid and rifampicin treatment increased lactate dehydrogenase release and CYP 3A4 activity significantly. Naringenin attenuated the effect of isoniazid and rifampicin on lactate dehydrogenase and CYP 3A4 activity.Conclusion Naringenin can attenuate the hepatotoxicity of isoniazid and rifampicin through inhibiting the activity of CYP 3A4 in cultured hepatocytes.

Objective To study effect of high ilioinguinal approach in treatment of acetabular fractures involving quadrilateral plate. Methods 28 cases of acetabular fractures were randomly divided into two groups according to admission time. 16 patients in the observation group were given high ilioinguinal approach operation. 12 patients in the control group were treated with traditional ilioinguinal approach op-eration. Operation condition, fracture reduction, the hip function recovery and postoperative complications of the two groups were compared. Results The amount of bleeding, the length of incision, operation time and operation conditions of the two groups had no significant differ-ence (P>0. 05). The satisfactory fracture reduction rate of observation group was higher than the control group (P<0. 05). Hip function score of observation group was significantly higher than that of the control group (P<0. 05). In observation group, the excellent rate of hip joint function was better than that of the control group (P<0. 05). The incidence rate of complications in the observation group and the con-trol group were 12. 50% and 16. 67% respectively, and there was no significant difference between the two groups (P>0. 05). Conclusion High ilioinguinal approach can improve the fracture reduction effect, effectively improve the hip joint function in patients, and it has higher security which is a kind of ideal treatment approach for acetabular fractures involving quadrilateral plate.

Objective To explore the role of VEGF positive expression in tumor tissue in the prognosis of liver transplantation for hepatocellular carcinoma (HCC).Method Fifty cases of liver transplant recipients with HCC confirmed immunohistochemically were enrolled in this study.The MaxVisionTM two-step method was applied to detect the expression of vascular endothelial growth factor(VEGF),and the microvessel density (MVD) was measured in para-cancerous tissues by using DAB staining.The correlation of the VEGF tumor tissue in tumor tissue with Child-Pugh,MELD,tumor diameter and number,differentiation,MVD,Milan criteria and UCSF criteria for HCC liver transplantation was analyzed.Result In the HCC tissue,the VEGF positive expression rate was 52％(26/50).The one-year survival of recipients positive and negative for VEGF was 78％ and 100％,respectively,and one-year recurrence rate was 32％ and 12％,respectively,with the difference being significant (P =0.043 and P =0.048 respectively).The expression of VEGF was associated with Child-Pugh,tumor diameter,MVD,Milan criteria and UCSF criteria (P＜0.05 for all).Logistic regression analysis showed that low differentiation and VEGF positive expression were independent prognostic factors for HCC recurrence after liver transplantation.Conclusion VEGF has a certain reference value to judge HCC invasiveness and prognosis of liver transplantation.

Aim To investigate the protective effect of cinnamic aldehyde ( CA ) on hormone-induced osteo-clasts proliferation and bone resorption in vitro and its molecular mechanisms. Methods RAW264. 7 cells induced into osteoclast were treated with RANKL and M-CSF and then were divided into control group, dexa-methasone ( DEX ) group and different doses of CA (11. 6, 23. 2, 46. 4 μg·L-1 ) groups. OCs were ob-served after tartrate resistant acid phosphatase( TRAP) staining. The cell proliferation was determined by MTT assay at different time points. The expression levels of TRACP5 b in cell cultured supernatants were measured by ELISA. RT-PCR technique was applied to examine the transcriptional levels of RANK and NFATc1 . Re-sults In MTT assay, the proliferation of osteoclasts stimulated by dexamethasone was promoted seriously compared with negative control group ( P < 0. 05 ) . Meanwhile, DEX could strengthen the content of TRACP5 b and up-regulate the expressions of RANK and NFATc1 mRNA. After administration of CA, the proliferation was inhibited, while the enhanced expres-sion of TRAP5b was reversed,and the over-expressions of RANK and NFATc1 mRNA were obviously down-regulated in a time-and-dose-dependent manner ( P <0. 05 ) . Conclusion The results suggest that CA in-hibits proliferation and bone resorption of osteoclast in-duced by DEX, which may be mediated by down-regu-lation of RANK and NFATc1 mRNA.

Objective To investigate the expressions of HBXIP and GRIM-19 in hepatocellular carcinoma tissues and their clinic significance. Methods Hepatocellular carcinoma tissue (n=42) and normal liver tissue (n=28) were collected from Tianjin First Central Hospital,immunohistochemistry was used to detect the expressions of HBXIP and GRIM-19 in these two groups. Results Rate of cells with positive expressions of HBXIP in hepatocellular carcinoma and normal liver tissues were 80.95%(34/42)and 42.86%(12/28)respectively;Rate of cells with positive expression of GRIM-19 in hepato?cellular carcinoma tissues and normal liver tissues was 40.48%(17/42)and 75.00%(21/28)respectively, and the difference between these two groups was statistically significant(P<0.05);The expression of HBXIP was higher but the expression of GRIM-19 was lower in poor differentiated and stageⅢ-IV cells than those in well and moderate differentiated cells and in stage I-II, cells. What′s more, the expression of GRIM-19 is higher in tissue without portal thrombosis than that in tissue with portal thrombosis. The expression of HBXIP was negatively correlated with GRIM-19 expression(rS=-0.400,P<0.01). Conclusion The abnormal expressions of HBXIP and GRIM-19 may play important roles in the process of development and metastasis of hepatocellular carcinoma.

ObjectiveTo study the role of middle hepatic vein (MHV) on the early function and regeneration of the donor remnant liver in living donor liver transplantation (LDLT).Methods Between August 2007 and August 2008,66 LDLT were performed,36 without MHV (group A),and 30 with MHV (group B) in the donor liver.The donor operation time,intraoperative blood loss,postoperative hospital stay,serum bilirubin,international normalized ratio (INR),alanine aminotransferase (ALT) and albumin were analyzed.We measured the volume of remnant liver with CT scan at 2 weeks after operation,and compared the function and regeneration of the remnant liver between the two groups. Results At 2 weeks after operation,there was no significant difference (P=0.16) in the volume of remnant liver between group A (959.3±195.2 ml) and group B (883.7±155.5 ml).There was also no difference (P=0.62) in the regeneration rate of segment IV between group A (78.2 ％ ± 29.1 ％) and group B (82.7 ％ ± 40.4％).The serum bilirubin,INR and ALT in group B was significantly higher than group A immediately after liver transplantation,but there was no difference at 1 week after transplantation.ConclusionExtended right hepatectomy with MHV was safe,and did not significantly impact early liver function and regeneration in the donor.

Objective To evaluate biliary digital imaging technology in determining the type of the intrahepatic bile duct anatomy and the transection plane of the duct in right lobe living donor liver transplantation(LDLT). Methods Mobile digital subtraction angiography was performed to show the intrahepatic bile duct anatomy of 66 liver transplant donor candidates. Combined with metal markers, the bile duct transection plane was defined. Comparing with the actual results, the effect of digital imaging technology in determining the intrahepatic anatomical variations and transection plane of the duct in LDLT was evaluated. Results Intrahepatic bile duct anatomical variations were showed in all donors by using digital imaging technology. type Ⅰ (classical type) was identified in45 cases (68.2%), type Ⅱ (with triple confluence, the simultaneous emptying of the right anterior segmental duct, right posterior segmental duct and left hepatic duct into the common hepatic duct) in 7 cases ( 10.6% ), type Ⅲ (no right hepatic duct stem, right posterior segmental duct draining into common hepatic duct) in 13 cases ( 19. 7% ), type Ⅳ (no right hepatic duct stem, right posterior segmental duct draining into left hepatic duct) in 1 case (1.5%), and type Ⅴ (complex variation ) in no case (0%). As a result, cases of type Ⅰ form a single anastomosis. In type Ⅱ, four cases formed double anastomoses, three cases formed single anastomosis with or without ductoplasty. In type Ⅲ, two anastomoses were formed in 9 cases, single anastomosis in 4 cases with ductoplasty. The case of type Ⅳ had double anastomoses. In all cases right lobe liver were harvested.Conclusions Biliary digital subtraction image combined with metal markers accurately defines intrahepatic bile duct anatomy and the transection plane, helping to reduce number of bile duct anastomosis, and contributes to safe graft harvesting.