ObjectiveTo study the effect of age on the perioperative and long-term outcome of hepatic resection for hepatocellular carcinoma. MethodsFifty two cases of elderly patients with hepatic resection for hepatocellular carcinoma were analysised retrospectively. ResultsThe morbidity rate and in-hospital duration in elderly group were 32.7% and (29.94.3)d respectively, higher than 18.6% and (24.76.1)d in non-elderly group (P

Objective To study the effect of age on the recurrence-free survival rate after hepatic resection for hepatocellular carcinoma(HCC)and the relationship between microvessel density (MVD)and recurrence of HCC in the elderly. Methods　Severty one cases of elderly patients with HCC were analyzed retrospectively with 352 cases of non-elderly HCC patients as control,and the effect of age on the recurrence-free survival rate was studied.The expressions of CD34 and endocan in HCC tissues were detected by immunohistochemistry in 30 elderly and 30 non-elderly patients.Results　The 1-,3- and 5-year recurrence free survival rates were 75.7%,43.0% and 43.0% in the elderly group respectively,which were higher than those in the non-elderly group(53.6%,38.5% and 33.4%,respectively,Log Rank value=10.25,P＜0.05).The positive rate of alpha fetoprotein (AFP)in the elderly group was 47.9%,which was lower than that in the non-elderly group(62.2%)(X2=23.68,P＜0.05).The median survival times in the high CD34-MVD group and high endocan MVD group were shorter than those in the low CD34-MVD group and low endocan-MVD group(260 d vs.850 d,360 d vs.800 d,Log Rank value was 22.18 and 20.56 respectively,both P＜0.05).Conclusions　The long-term prognosis of hepatic resection for HCC is better in elderly patients than in non-elderly patients.The recurrence of HCC in the elderly is closely related with angiogenesis.

ObjectiveTo investigate the expression of RhoC gene in primary hepatocellular carcinoma and to evaluate the relationship between RhoC gene expression and invasion and metastasis of primary hepatocellular carcinoma.MethodsThe mRNA expression of RhoC gene was examined by polymerase chain reation after reverse transcription (RT-PCR) in 25 cases of primary hepatocellular carcinoma (HCC) and adjacent non-cancerous tissuse. In addition, the mutation of RhoC gene was examined by polymerase chain reaction-single strand conformational polymorphism(PCR-SSCP)ResultsThe mRNA expression of RhoC in tumor tissue were higher than that in adjacent liver tissue,1.8?1.1 vs. 1.0?0.7( P

Objective To investigate the expressions of vascular endothelial growth factor (VEGF) , hypoxia inducible factor 1 alpha (HIF 1?) and epidermal growth factor (EGF) in hepatocellular carcinoma (HCC) and their clinical significance. Methods The expressions of VEGF, HIF 1? and EGF in 36 cases of HCC and corresponding paraneoplastic tissues and normal liver tissues (6 cases) were studied by immunohistochemistry assay. ResultsThe expression rate of VEGF, HIF 1? and EGF in HCC tissue was 89%, 67% and 75% respectively, higher than those in paraneoplastic tissues and normal liver tissues ( P

BACKGROUND: Through exercise and/or hypoxia to increase the body's stress level and timing of hypoxia, so as to improve the body's adaptation level to exercise and/or hypoxia. However, little was known concerning the effects of acute exercise and/or hypoxia on vascular endothelial cell growth factor (VEGF) expression in skeletal muscles.OBJECTIVE: To study the effects of acute exercise and/or hypoxia on VEGF expression in rats' gastrocnemius muscles. DESIGN, TIME AND SETTING: The randomized controlled animal experiment was performed at the Clinical Laboratory, People's Hospital of Jiangsu Province, between September 2005 and September 2006.MATERIALS: Totally 108 health male SD rats were randomly divided into 6 groups, namely, normoxia quiet, normoxia high intensity, normoxia moderate intensity, hypoxia quiet, living high-training low high intensity and living high-training low high intensity moderated intensity groups, with 12 animals in each group.METHODS: In acute normoxia exercise models, rats were performed adaptive activity at 48 hours prior to experiment. The high intensity exercise was comprised of 50 m/minx1.5 min training with 2 minutes rest. The moderate intensity exercise was 30 m/min×30 min. Hypoxia environment was produced by using low oxygen instrument to simulate hypoxia training, with hypoxia for 3 days, 22 h/d, 12.8% altitude, with 22 ℃ temperature and 55% humidity. In acute training low-living high models, rats were placed in above hypoxia environment after high intensity Or moderate intensity exercise. Four rats were sacrificed at hours 0, 2 and 4 after training, and the gastrocnemius muscles were obtained.MAIN OUTCOME MEASURES: The expression of VEGF in rats' gastrocnemius muscles was detected by using western-blot.RESULTS: Hypoxia and acute normoxia exercise enhanced the expression of VEGF, hypoxia after exercise weakened exercise-induced VEGF expression, and the exercise with long time and common intensity induced the higher level VEGF expression. The expression of VEGF was the most at the time points of instantaneousness and 2 hour after exercise, the sorting of the recovery speed of VEGF changes from fast to slow was: hypoxia or training low-living high and normoxic exercise. CONCLUSION: The expressions of VEGF in rats' skeletal muscles induced by acute exercise and/or hypoxia belong to the effect of immediate-early, with existing intensity-threshold, which recovery speed is inversely proportional to the expression amplitude;"training low-living high" may be able to enhance the adaptation of skeletal muscles to sports.

ObjectiveTo study the expression of microRNA-21 ( miR-21 )in serum of patient with diffuse large B cell lymphoma (DLBCL) and DLBCL cell lines and validate the significance of miR-21 in early diagnosis,genotyping and prognosis estimates of DLBCL.MethodsmiR-21 expression were detected by fluorescent quantity polymerase chain reaction (FQ-PCR)in 9 lymphoma cell lines (OCI-Ly1,OCI-Ly3,OCI-Ly4,OCI-Ly7,OCI-Ly8,OCI-Ly10,OCI-Ly18,OCI-Ly19 and HBL),the serum from DLBCL patients (n =62) and health controls (n =50 ).Kaplan-Meier survival analysis was carried out during the relapsefree survival period of DLBCL patients to explore the relationship between the prognosis and microRNA expression level.ResultsReal time FQ-PCR result indicated that miR-21 expression was higher in DLBCL cell lines than that in normal B cells (BC).miR-21 expression in normal B cell and 9 DLBCL cell lines separately were 1.04 ± 0.02,2.30 ± 0.35,237.97 ± 56.19,5.27 ± 0.83,3.40 ± 0.30,11.22 ± 2.70,133.55 ± 16.78,6.63 ±0.24,4.91 ±0.37 and 81.59 ±6.64.Compared with BC,the expression of miR-21 were higher in all 9 DLBCL cell lines ( t =7.3,13.7,21.0,6.2,8.8,13.6,6.5,39.5,18.1 ;P ＜ 0.01 ).miR-21 expression segregates with specific molecular subgroups of DLBCL The expression was higher in the ABC type cell lines (OCI-Ly3,OCI-Ly10,HBL) than GCB type cell lines (OCI-Ly1,OCI-Ly4,OCI-Ly7,OCI-Ly8,OCI-Ly18,OCI-Ly19;t =11.18,P ＜ 0.01 ).Consistent with the cell line models,miR-21 expression levels were higher in serum from DLBCL patients [21.38 (10.26-45.21 )] than from controls [1.87 ( 1.05-3.97 ),U =168,P =0.000],and the levels were higher in DLBCL cases with an ABC-type [28.68 ( 14.92-98.44 )] than those in GCB-type [18.30 ( 7.32-33.46 ),U =336,P =0.043].MiR-21 expression levels were different in sera from different clinical stage DLBCL patients.The miR-21 level in serum of patients with subgroup ABC and subgroup GCB in stage Ⅰ and Ⅱ were 47.49( 25.65-295.41 ) and 24.74( 16.08-50.38) respectively and in stage Ⅲ and Ⅳ were 16.66 ( 5.35-44.30 ) and 11.96 ( 4.10-21.05) respectively.The levels were higher in DLBCL cases withⅠ -Ⅱ stage than those with Ⅲ-Ⅳ stage (U =62,P =0.013 in GCB type; U =53,P =0.014 in ABC type).Moreover,compare with relapse-free survival in DLBCL patients,high miR-21 expression was associated with well prognosis ( U =259,P =0.035).ConclusionsMiR-21 is high expression in DLBCL cell lines and DLBCL patients serum.miR-21 level in sera from DLBCL patients is associated with clinical stage,molecular subgroup and prognosis estimates.MiR-21 may serve as a new biomarker to early detection,genotyping and prognosis estimates of DLBCL.

Objective To explore the inhibitory effect and possible mechanisms of lianhuaqingwen capsules on radiation-induced acute lung injury in rats. Methods Rats were randomly divided into control group, radiation group and radiation plus lianhuaqingwen group, the control group and the radiation group rats were given 0. 9% sodium chloride solution, the radiation plus lianhuaqingwen group rats were given lianhuaqingwen 0. 9% chlorine sodium solution. HE staining was applied to test the lung tissue inflammation; quantitative RT-PCR and ELISA were used to measure the content of IL-6, TNF-α and MCP-1 in rats;immunohistochemical assay was taken to detect the infiltration of macrophage in lung tissues. Results The relative mRNA expression of IL-6, TNF-α and MCP-1 in the control, radiation model control and radiation plus Lianhuaqingwen groups were (0. 002 1±0. 000 20),(0. 006 6±0. 000 32),(0. 003 9±0. 000 22); (0. 003 7±0. 000 16),(0. 007 4±0. 000 33),(0. 005 5± 0.000 24);(0.001 4±0.000 15),(0.005 4±0.000 72),(0.003 2±0.000 17),respectively; the concentration (pg·mL-1) of IL-6,TNF-αand MCP-1 in the serum were (35. 2±10. 9),(111. 8±26. 1),(68. 2±15. 2); (229. 3±28. 5),(837. 5±57. 6), (566. 9±39. 8);(96. 85±8. 20),(314. 53±12. 76),(191. 32±10. 97),respectively; and the macrophages at high magnification field in each group were (59. 5±4. 3),(503. 9±25. 8)and (106. 2±12. 6), respectively. Lianhuaqingwen capsules significantly alleviated the lung inflammation in rats with radiation-induced acute lung injury,inhibited the accumulation of macrophage in lung tissue,reduced the expression of IL-6 and TNF-α,and decreased the content of MCP-1 in lung tissues and sera(P<0. 05). Conclusion Lianhuaqingwen capsules attenuated the lung inflammation developed in rats with radiation-induced acute lung injury through inhibiting the expression of MCP-1 and reducing the accumulation of macrophage in lung tissues.

Objective To investigate the radioprotective function of lianhuaqingwen (LHQW) in rat acute radiation-induced lung injury.Methods Totally 36 female Wistar rats were randomized into 3 groups as administered group (treated by LHQW plus radiation),radiation group irradiated with a single of 20 Gy in 6 MV X-ray by Elekta Synergy VMAT,and blank control group without radiation.Performance status (PS) was estimated during 31 d of LHQW instragastric administration.After rats being sacrificed at 1,14,28 d of LHQW adminstration,the pathomorphological changes were observed in trauma lung tissue,the cell number in BALF (Bronchoalveolar lavage fluid) was counted,the levels of TNF-α and IL-6 in serum were measured by ELISA,and TNF-α and IL-6 mRNA expressions in lung tissue were assayed by RT-PCR.Results After LHQW treatment,the PS of rat was significantly elevated with less inflammation in morphous,and the cell number in BALF was markedly decreased in compare with radiation alone group.Furthermore,the serum levels of TNF-α and IL-6 were obviously reduced (tTNF-α =7.372,2.891,tIL-6 =6.335,3.257,P ＜ 0.05) and the TNF-α and IL-6 mRNA levels in lung tissue were also decreased (tTNF-αmRNA =3.714,2.144,tIL-6mRNA =3.589,2.883,P＜0.05).Conclusions LHQW plays a protective role against acute radiation-induced lung injury in rats and the down-expressions of TNF-α and IL-6 may be involved.

Objective To investigate the expression level of microRNA-145 in breast cancer cell lines andtissues and its impact on breast cancer invasion and metastasis.Methods MiR-145 expression was detected by FQ-PCR in 5 breast cancer cell lines ( HBL-100, MCF-7, MDA-MB-231, MDA-MB-468 and SK-BR-3)and in breast cancer tissue and paraneoplastic tissues (n=39).The miR-145 expression plasmid ( Psif-miR-145 ) and negative control plasmid were transfected into SK-BR-3 using lipofectamine, respectively.The characteristics of invasion and migration of the transfected SK-BR-3 cells were examined by scratch test and transwell assay.The target genes of miR-145 were predicted by bioinformatics and the ANGPT2 gene were verified as miR-145 target by the dual-luciferase reporter assay.The expression levels of ANGPT2 protein was examined by western blot after pSIF-miR-145 transfection by lipofectamine in breast cancer cell line SK-BR-3.Results FQ-PCR result indicated that miR-145 expression level waslower in breast cancer tissue (45.93 ±22.02)than paraneoplastic tissue [ (182.04 ±56.92), U value was 7, P<0.01].MiR-145 expression level was lower in breast cancer cell lines than normal breast cells.miR-145expression in 4 breast cancer cell lines was 0.51 ±0.05, 0.07 ±0.01, 0.36 ±0.04 and 0.04 ±0.01, respectively.Compare with normal breast cell, miR-145 was lower expressed in all 4 breast cancer cell lines (t value separately was 15.93, 308.17, 25.02, 201.30;P<0.05).Lower expression of miR-145 was observed in the highly invasive breast cancer cells (MDA-MB-231, MDA-MB-468 and SK-BR-3), compared with weakly invasive breast cancer cell (MCF-7) (t value separately was14.18, 3.78, 15.20;P<0.05). Wound healing assay shows that overexpression of miR-145 in SK-BR-3 significantly reducedthe motility as compared with control group (P <0.01).The cell invasion assay indicated the numbers of miR-145 overexpressed SK-BR-3 cells, which invased to lower chamber, was 137 ±37, the numbers of invased cells was 617 ±80 when the negative control was applied. Over-expression of miR-145 could repress the expression levelsof ANGPT2 protein;miR-145 could repress the activity of luciferase reporter carrying a 3′-untranslated region of ANGPT2 mutated the predicted binding site, the activity of luciferase was reversed. Conclusions MiR-145 depressed in breast cancer cell lines and breast cancer tissues.MiR-145 maybe plays an important role in breast cancer invasion and migration by directly target ANGPT2.

Objective :To study a simplified method of isolation of rat hepatocytes and to observe the pro-cess of cell morphology in long-term culture. Methods :Rat hepatocytes were isolated by a single two-stepperfusion method. The yield and viability were assessed by trypan blue exclusion. [3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide] (MTT) was used to test the effect of serum concentration of newborn calf serum on the proliferation of hepatocytes. Hepatocytes were inoculated in the culture mediumconsisted of Williams' E supplemented with insulin,dexamethasone and 10％ new born calf serum. Themorphologic change of cultured hepatocytes was observed. Results:The average yield of hepatocytes was 2.26× 108 cells per rat, with an average viability of 95.6％. New born calf serum had strong biological activi-ty to stimulate the proliferation of hepatocytes and there was close-effect relationship followed by the in-crease of new born calf serum concentration. The rat hepatocytes can be cultured for 5～ 6 weeks withpreservation of normal morphologic appearance. Conclusion:The rat hepatocytes isolated by the abovemethod have high yields and viability and can be long-term cultured in vitro.