Objective To evaluate the safety and feasibility of hepatic resection for huge primary liver carcinoma (PLC). Methods 216 cases of huge PLCs(mean diameter of 14.2cm) were resected. The hepatectomies were performed under intermittent occlusion of hepatic inflow. Results All 216 cases were successfully resected. The mean time of occlusion of hepatic inflow was 19min, the mean blood loss was 743 ml. No serious complications occurred, and only seven patients died of hepatic failure and upper gastrointestinal haemorrhage postoperatively in this series. Conclusions Although resection of huge PLC is quite difficult, but if suitable surgical techique and perioperative management are adopted ,it is safe and feasible .

Objective To investigate the relationship between the co-expression of CD105 and cyclin D1 and lymph node metastasis and prognosis of esophageal squamous cell carcinoma ( ESCC ). Methods Eighty cases of ESCC tissue were collected at The First People's Hospital of Nantong. The expression of CD105 and cyclin D1 of ESCC was detected by immunohistochemistry. The relationship between the co-expression of CD105 and cyclin D1 and lymph node metastasis and prognosis of ESCC was analysed. Eighty normal esophageal tissues were selected as controls. All data were analysed by t test, chi-square test, and Pearson correlation analysis. Survival was analysed by the Kaplan-Meier survival curve. Microvessel density (MVD) was used to indicate the expression of CD105 in the form of -x±s. Results The expression of CD105 in ESCC tissues was higher (36±8) than that in normal esophageal tissues ( 11±3) (t =25. 129, P＜0.05). The positive rate of cyclin D1 expression in ESCC tissues was 61% (49/80), which was significantly higher than 23% (18/80) in normal esophageal tissues ( x2 =4.972, P＜0.05). The MVD value of 44 patients was ≤36 (LCD105), and nine of them had lymph node metastasis. The MVD value of the remaining 36 patient was ＞ 36 ( HCD105 ), and 26 of them had lymph node metastasis. Twenty-eight patients with positive expression of cyclin D1 had lymph node metastasis, while seven patients with negative expression of cyclin D1 had lymph node metastasis. The results of Pearson correlation analysis revealed that a high expression of CD105 and a positive cyclin D1 expression were correlated with lymph node metastasis (x2 =21.562, 9.217, P＜0.05). The survival times of 28 patients with positive cyclin D1 and HCD105,21 patients with positive cyclin D1 and LCD105, eight patients with negative cyclin D1 and HCD105, and 23 patients with negative cyclin D1 and LCD105 were (31±6) months, (47±7) months, (51±9) months and (61±5) months, respectively, with a significant difference among the four groups (F = 11.76, P ＜ 0. 05 ).Conclusion Co-expression of CD105 and cyclin D1 may be used as a prognostic factor of ESCC.

Objective To explore the damage of spiral ganglion neurons(SGNs),the protective effects of different dosages of healthy ear compound (HEC)from traditional Chinese medicine(TCM) against age-induced SGNs degeneration and its possible mechanism in spiral ganglion neurons(SGNs)of C57BL/6J mice.Methods Totally 36 C57BL/6J mice just after ablactation were randomly divided into four groups.Normal control group (n =6)drank tap water daily from ablactation to 2 months old.Ageing-related SGNs apoptosis control group(n=12)drank tap water daily from ablactation to 7 months old.High dose TCM group(n=12)at drank 3.65 g/kg/d of HEC from ablactation to 7 months old.Low dose TCM group(n=6)drank 0.91 g/kg/d of HEC from ablactation to 7 months old.The animal cochleae were immediately removed at the termination of the experiment.In each group,the cochleae of 6 animals were used for paraffin embedding,slicing and toluidine blue staining to observe neuronal morphological changes.The caspase-3 mRNA expression study was performed by real-time PCR technique in 6 cochleae of High dose TCM group and ageing-related SGNs apoptosis control group.Results The morphological structure of cochlear SGNs represented healthy and normal density in normal control group at 2 months old.In contrast,amount or density of SGNs in cochlear basilar part was significantly damaged and reduced in ageing-related SGNs apoptosis control group at 7 months old(P＜ 0.001).But the high dose TCM group at 7 months of age was similar to the normal control group at 2 months old in morphological structure,amount or density of SGNs(P＞0.05).The low dose TCM group was significantly different from other 3 groups in amount or density of SGNs (P＜0.001).However,SGNs in the middle part and apical part showed integrity in each group.In addition,the expression level of caspase-3 in the cochlea of high dose TCM group was also obviously different with age-related SGNs apoptosis control group(P＜0.01) Conclusions Ageing-related damage of SGNs in C57BL/6J mice begins from the base of cochlea and progresses towards the apex.The HEC of TCM could significantly protect SGNs against age-induced apoptosis in SGNs.The efficacy of the high dose TCM is better than that of the low dose TCM.Its SGNs protective mechanisms might be related to involving the caspase-mediated cell apoptotic pathway.

Objective To clone and efficiently express E6E7 proteins of human papilloma virus HPV33 in E.coli.The recombinant proteins were purified and immunized to mice, and polyclonal antibodies were purified.Methods E6 and E7 genes of HPV33 were amplified by PCR using HPV33 human papilloma virus as template, and were sub-cloned into pET28a and pCDNA3.1 /His C vectors.The recombinant plasmids pET28a-HPV33 E6 and pET28a-HPV33 E7 were transformed into Escherichia coli BL21 (DE3) strain.We purified the recombinant proteins and immunized mice for preparing antibodies, and the polyclonal antibodies were purified. Moreover the plasmids pCDNA3.1 /His C-HPV33 E6 and pCDNA3.1 /His C-HPV33 E7 were transfected into 293 cells.The cells were used in immunofluorescence and Western-Blot to detect antibody specificity. Results The recombinant proteins were successfully obtained, and the polyclonal antibody was prepared, which had good specificity. Conclusion The recombinant proteins and antibodies may be used in the clinical detection and pathogenesis research of HPV33 type human papilloma disease.

OBJECTIVETo determine the effects of ginsenosides Rb1(GSRb1) on learning and memory and expression of somatostatin (SS) in the hippocampus and the frontal cortex in rat model of sleep deprivation (SD).

METHODSRats were randomized into groups of SD 2 d, SD 4 d, SD 6 d, and SD 0 d, while each group was sub-divided into GSRb1 group and normal saline (NS) sub-groups. Rats were intraperitoneal administered with 30 mg/(kg*d) of GSRb1 or NS for 7 d, then the learning and memory abilities were examined by measuring average swimming speed and mean escape latency using Morris maze.Expression of somatostatin was detected with immunohistochemical method and image analysis in the hippocampus and the frontal cortex.

RESULTSCompared with SD 0 d rats, SD rats exhibited significant decrease in the average swimming speed and increase in the escape latency (P <0.01). The expression of somatostatin in the hippocampus was decreased significantly in SD 2 d, SD 4 d and SD 6 d rats (P<0.05). However, decrease was only observed in SD 4 d and SD 6 d rats in the frontal cortex (P <0.05). Parallel comparison between NS control and GSRb1 treated rats demonstrated that rats treated with GSRb1 in each subgroup exhibited faster swimming speed and shorter escape latency (P <0.05). Meanwhile, the expression of somatostatin was increased in SD 2 d, SD 4 d and SD 6 d rats in the hippocampus and in SD 4 d and SD 6 d rats in the frontal cortex (P <0.05), respectively.

CONCLUSIONGSRb1 enhances the expression of somatostatin in sleep deprivation rats and subsequently may improve learning and memory abilities of rats.

Animals ; Brain ; metabolism ; Disease Models, Animal ; Ginsenosides ; pharmacology ; Learning ; drug effects ; Male ; Memory ; drug effects ; Rats ; Rats, Sprague-Dawley ; Sleep Deprivation ; metabolism ; Somatostatin ; metabolism

OBJECTIVETo study the effects of sodium butyrate (NaBu) on cell cycle checkpoint and the apoptosis sensitivity in U937 cells.

METHODSTwo mutant U937 cell lines, U937-ASPI3K (ATM negative) and U937-pZeosv2(+) (ATM wild-type), were used as the cell model system. Immunoprecipitation and kinase assay were used to examine the p38 MAPK and ERK1 kinase activities. Western blot was used to analyze the phosphorylation of Bad protein.

RESULTSU937-pZeosv2(+) pretreated with NaBu exhibited enhanced apoptotic response in a NaBu dose dependent fashion upon (137)Cs irradiation, which could be abolished by olomoucine (OLM), a p38 MAPK specific inhibitor. On the other hand, Cyclin dependent kinase 2 (CDK2) specific inhibitor CDK2-I and p34cdc2/cyclinB inhibitor alsterpaullone (ALP) failed to block the effects of NaBu. Similar results were also observed in U937-ASPI3K. The effect of irradiation on p38 MAPK and ERK1 was strikingly potentiated by NaBu. Furthermore, inactivation of irradiated Bad protein via phosphorylation on serine 136 was also enhanced.

CONCLUSIONNaBu is able to enhance the apoptotic response in U937 cells, which is mediated by p38 MAPK activation but not ATM status.

Apoptosis ; Butyrates ; pharmacology ; Carrier Proteins ; metabolism ; Humans ; MAP Kinase Signaling System ; Mitogen-Activated Protein Kinase 3 ; Mitogen-Activated Protein Kinases ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; U937 Cells ; bcl-Associated Death Protein ; p38 Mitogen-Activated Protein Kinases

Objective To explore the epidemiological status of abnormal glucose metabolism and its influential factors among middle and aged population with hypertension in Chengdu area. Methods In 2008, after adopting the methods of stratified cluster sampling, the authors investigated 4685 subjects of the middle and aged population between the age of 40-79 in Chengdu urban and rural area by checking blood pressure and oral glucose tolerance test (OGTY). Patients with previously known diabetes mellitus (DM) were only asked to perform fasting glucose and to carry out a questionnaire. Comparison of the prevalence rates of abnormal glucose metabolism in hypertensive and non-hypertensive subjects was carried out. The prevalence rates of isolated impaired glucose tolerance (I-IGT) and isolated postprandial hyperglycemia (IPH) among middle and aged subjects with hypertension were acquired and the influential factors of abnormal glucose metabolism among middle and aged subjects with hypertension were analyzed. Results The prevalence rate of abnormal glucose metabolism in the hypertensive subjects was obviously higher than that in the non-hypertensive subjects; without using OGTT, 72.9% of the pre-diubetic and 54. 4% of the new diagnosed DM patients would remain undiagnosed if fasting plasma glucose detection was used alone. Age, diabetic history of first degree relatives ,overweight or obesity were the risk factors for the development of abnormal glucose metabolism among middle and aged male subjects with hypertension in Chengdu area. Exercise training and high education level were the protective factors. Age, diabetic history of first degree relatives,abdominal obesity and hypertriglyceridemia were the risk factors for the development of abnormal glucose metabolism among middle and aged female subjects with hypertension in Chengdu area. Conclusions More than 50% of middle and aged subjects with hypertension in Chengdu area has accompanying abnormal glucose metabolism. OGTT easily discloses the abnormal status and should be a routine procedure in the diagnosis of pre-diabetes or DM in such population. Appropriate exercise, learning diabetes-related knowledge to take reasonable lifestyle, and intervention of metabolic factors such as overweight or obesity are advised. Abdominal obesity and hypertriglyceridemia play important roles in leading to abnormal glucose metabolism among middle and aged population with hypertension.

In order to explore the role of BspE protein in Brucella infection,yeast two-hybrid tech-nique was used to screen host cell proteins that interact with BspE protein.The constructed BspE recombinant plasmid pGBKT7-BspE was used as bait plasmid to hybridize with the RAW264.7-cD-NA library of mouse mononuclear macrophages by yeast two-hybridization technique.The positive clones were extracted by plasmid,sequenced and co-immunoprecipitation to determine the host cell proteins that could interact with BspE.The subcellular localization of BspE proteins was analyzed by confocal laser microscopy.The physical and chemical properties,protein structure and function of BspE interacting proteins were analyzed by bioinformatics.The siRNA for one of the BspE inter-acting proteins was synthesized,the expression of its gene was silenced in HEK293T cells,and the silenced cells was infected with Brucella M5-90 and the number of intracellular bacteria was coun-ted.The results showed that the decoy plasmid pGBKT7-BspE was successfully constructed,and the plasmid could express BspE protein in yeast.Eight positive clones were obtained from the host cell genome library by yeast two-hybridization.The positive clones were identified as RBM27 and PCBP1 by sequencing,backcross and co-immunoprecipitation.Bioinformatics was used to predict the cell location,protein structure and amino acid composition of RBM27 and PCBP1.After siRNA interference,the expression level of PCBP1 was significantly decreased and the amount of M5-90 in the cell was increased.Brucellosis secreted protein BspE interacts with host proteins RBM27 and PCBPl,and PCBP1 negatively regulates the proliferation of Brucellosis.

Objective To investigate the prokaryotic expression and immunoreactivity of BspE,a type Ⅳ secretion protein of Brucella,and the effect of recombinant protein BspE on cytokines.Methods According to the BspE gene of Brucella M5-90 published in GenBank,the gene fragments were synthesized by a company and then ligated into PUC57 vector for sequencing.The sequenced gene was cloned into a prokaryotic expression vector pET-28α and transformed.Induced expression was performed in E.coli DE3 competent cells.The obtained target protein was purified by a Ni-NTA affinity column,and its reactogenicity was analyzed by Western blotting.Mouse RAW264.7 cells were treated with 25 g/L BspE recombinant protein for 12,24,48 h,and the control group was treated with the same amount of BSA instead of BspE,and enzyme-linked immunosorbent assay (ELISA) was used to detect interleukin (IL)-1β level.Results The recombinant expresed plasmid of pET-28α-BspE was successfully obtained.The results of Western blotting showed a single band with a relative molecular mass of about 30.1 × 103,and the recombinant protein BspE had good reactogenicity,and IL-1β levels (ng/L)were significantly elevated by the recombinant protein BspE (12 h:43.27 ± 2.13 vs 30.24 ± 1.66,24 h:57.78 ± 3.44 vs 41.22 ± 1.22,48 h:72.52 ± 3.04 vs 46.77 ± 2.75,t =8.38,7.86,10.89,P ＜ 0.05).Conclusions BspE recombinant protein has better immunoreactivity and can increase the expression level of IL-1β in mouse macrophages.This study provides a scientific basis for the role of effector proteins in the pathogenesis of Brucella.

AIM:We investigated how AT 1-R stimulated by mechanical stresses induces cardiac fibrosis .METHODS:We produced in vivo cardiac pressure overload model in angiotensinogen knockout ( ATG-/-) mice and in vitro mechanically-stretched cell model in cultured neonatal cardiac cells of ATG-/-mice both lack the participation of Ang II .RESULTS: Pressure overload for 4 weeks in ATG-/-mice induced myocardial hypertrophy accompanied by the significant interstitial fibrosis , however , the TGF-β, a key regulatory factor of fibrosis, was not significantly increased in these ATG-/-mice.Meanwhile, the inhibitor for AT1-R significantly inhibited mechani-cal stress-induced cardiac fibrosis in these ATG-/-models whereas inhibition of TGF-βdid not.CONCLUSION:The results showed that mechanical stress-induced fibrotic responses through AT 1-R required the phosphorylation of Smad 2 but not the involvement of TGF-β.