Objective To investigate the anticancer activity of dendritic cells(DC)transfected by recombinant adeno-associated virus(rAAV)-mediated carcinoembryonic antigen(CEA)gene in colorectal cancer treatment.Methods The recombinant plasmid rAAV/CEA was constructed and the rAAV/CEA virus stock was prepared.DC precursors were isolated from peripheral blood mononuclear cells(PBMC)by density gradient centrifugation and transfected with rAAV/CEA virus stock.The CEA expression,chromosome integration and efficiency of rAAV/CEA virus were determined by PCR,Southern blot and flow cytometery.DC precursor cells were induced by granulocyte macrophage colony-stimulating factor(GM-CSF),interleukin-4(IL-4)and tumor necrosis factor-?(TNF-?)to mature DC.The mature DC and T cells were mixed,then cultured with interleukin-2(IL-2)and interleukin-7(IL-7)resulting in generation of cytotoxic T lymphocyte(CTL),whose lethal effects on cancer cells were determined by 51chromium(51Cr)release assay.3H incorporation method was employed to detect the influence of rAAV/CEA transfected DC in the proliferation of activated T cells.Results rAAV medicated CEA gene was transferred into DC precursor successfully and the expression was high.Over 90% DC precursors expressed CEA protein.The viral gene was integrated into the DC chromosome.rAAV/CEA transfected DC could induce specific CTL which could specifically recognize and kill CEA-positive colorectal cancer LoVo cell line,and showed restrictive activity of anti-MHC class I antigen,while it was not able to kill the CEA-negative cancer cells.Conclusion The present study confirms that rAAV/CEA transfected DC vaccine can induce anti-tumor activity in vitro.It paves the way for further study on CEA as a target antigen for gene therapy on colorectal cancer.

Objective Epidermal growth factor receptor (EGFR) plays an important role in the pathogenesis of colorectal cancer (CRC). There are several potential therapies to target EGFR. The detection of EGFR expression is usually performed by immunohistochemistry (IHC) only in the primary tumor. There are a few data regarding the EGFR status in the corresponding distant metastases. The aim of the present study was to correlate EGFR expression in primary tumors and related metastases in order to find out whether EGFR status is different between primary tumors and related metastases. Methods 37 cases (25 male, 12 female) of metastatic colorectal cancer were collected. The site of primary tumor was on colon in 16 patients (43.2%) and on rectum in 21 patients (56.8%). Metastatic sites analyzed were liver in 32 patients (86.5%), inguinal lymph nodes in 5 patients (13.5%). The EGFR status of primary tumors and related metastatic sites were evaluated retrospectively. The expression of EGFR was analyzed by immunohistochemical staining (IHC), EGFR status was defined as positive if the percentage of stained malignant cells was ≥5%, and the expression feature was analyzed. Results EGFR status was positive in 25 primary tumors (67.6%), in which the corresponding metastatic site was negative in 7 cases (28.0%), whereas it was positive in 3 metastases (25.0%) from 12 cases of EGFR-negative primary cancers. The difference between the primary tumors and metastatic lesions had no statistic significance (P=0.23). Conclusion When analyzed by IHC, The difference of EGFR status has no statistic significance between the paired primary tumors and distant metastatic lesions, and 73% of samples show concordant EGFR status.

Objective To investigate the application of expanded p01ytetrafluoroethylene(ePTFE)grafts in upper arm to build arteriovenous aCCeSS for hemodialysis. Methods ePTFE graft vascular access was built in the upper arm in 20 uremia patients.Three operation strategies were applied according to the reference,including loop grafts connected axillary artery and axillary vein,straight graft connected axillary artery and elbow basilic vein,and bridge connected elbow brachial artery and axillary vein. Results Twenty operations were successful and after 6-8 weeks the fistula of all cases were used in hemodialysis.The blood flows were 220-300 ml/min without re-circulation found.Conclusion ePTFE graft arteriovenous vascular access in the upper arm could be an alternative for hemodialysis patients who are difficult to build native arteriovenous fistula.

Objective To find the normal range and features for auditory brainstem responses(ABR)of infants within six months,and the basic parameters for identifying the hearing impaired infants.Methods May 2006 to May 2008,60 normal infants and 20 normal adults received the ABR tests.The infants were divided into three groups:group A(6 weeks),group B(3 months)and group C(6 months).Each group consisted of 20 infants(40 ears)and the ABR data were statistically analyzed.Results At 100 dB nHL,the waveⅠ,Ⅲ,Ⅴ latencies of ABR for 6-week infants respectively were 1.49?0.08 ms,4.42?0.16 ms and 6.61?0.25 ms,for 3-month infants,1.47?0.07 ms,4.35?0.20 ms and 6.50?0.25 ms,for 6-month infants,1.45?0.07 ms,4.17?0.15 ms and 6.32?0.22 ms.At 80 dB nHL,the waveⅠ,Ⅲ,Ⅴ latencies of ABR for 6-week infants were 1.63?0.08 ms,4.52?0.17 ms and 6.74?0.26 ms,respectively,for 3-month infants,1.64?0.11 ms,4.44?0.20 ms and 6.67?0.26 ms,for 6-month infants,1.60?0.11 ms,4.27?0.16 ms and 6.43?0.24 ms.As the intensity of stimulating signals decreased,the peak latency of ABR became more delayed.The peak latencies of Ⅲ,Ⅴ waves and interval peak latencies of Ⅰ-Ⅲ and-Ⅴ waves for the infants were more delayed than those for adults,and became shorter with the increase of the infants' ages,but they failed to reach adults' levels even for 6-month infants.However,the length of wave Ⅰ of ABR for infants of different months was close to that for adults.The thresholds of ABR for the infants of different ages were not significantly different,all being close to the values for adults.Conclusion This study has analyzed the ABR wave features for the infants of three ages and this can help to define the reference ranges for the infants and to provide the parameters for early diagnosis of hearing loss among the infants.

objective To investigate the effects of different sections of receptor associated protein (RAP) on the expression and distribution of TRPC6,synaptopodin and podocalyxin in passive Heymann nephritis(PHN). Methods Male Sprague-Dawley rats were injected with three kinds of antisera (anti-RAP full-length serum,anti-RAP N-terminal serum and anti-RAP C-terminal serum)to establish three kinds of PHN models.The control group was injected with normal rabbit serum.The quatitation of 24 h urinary protein,serum albumin and creatinine were taken before injection and one week after PHN model successfully induced.The histopathologic changes of renal tissues were observed by light microscopy.The expression and distribution of TRPC6,synaptopodin and podocalyxin in glomerular podocytes were observed by laser scanning confocal microscopy and analyzed by fluorescence quantitative software after indirect immunofluorescence double staining.Results The quantities of 24 h urinary protein in the three model groups were significantly higher than those of themselves before injection and control groups (P<0.01,respectively).The values of serum albumin and creatinine were not significantly different before and after injection (P>0.05).The expression of TRPC6 in podocytes was higher in the PHN model groups than that of control group.Fluorescence intensity of TRPC6 in RAP full-length group was stronger than that in RAP N-terminal or C-terminal groups.The expressions of synaptopodin and podocalyxin distributed along the glomerular basement membrane as spot,discontinuous short line and defect of some segments,and were lower in three PHN groups than those of control group.Fluorescence intensity of synaptopodin and podocalyxin among three PHN groups had no differences. Conclusions RAP full-length and N-terminal or C-terminal parts can increase the expression of podocyte TRPC6,but decrease the expressions of synaptopodin and podocalyxin,and alter their distribution,which may be associated with the proteinuria,however,their role in the PHN pathogenesis needs further study.

Objective To investigate the diagnosis and treatment of anaplastic carcinoma of the pancreas.Methods The clinical data of 10 patients with anaplastic carcinoma of the pancreas who were admitted to the Zhongshan Hospital from January 1999 to June 2010 were retrospectively analyzed.Computed tomography was carried out preoperatively and surgical plan was designed according to the site of tumors.Chemoradiotherapy was applied postoperatively.Patients were followed up till March 2012 by phone call and out-patient examination.The clinical and imaging features,pathological characteristics,treatment and follow-up data were analyzed.The clinical and pathological features were analyzed by descriptive statistics.The continuous data were described as (x) ± s,and categorical data were presented by frequency and precentage.Results The tumors located at the head of the pancreas were observed in 5 patients,tumor located at the neck of the pancreas in 1 patient,and tumors located at the body and tail of the pancreas in 4 patients.Two patients underwent pancreaticoduodenectomy (PD),2 underwent PD + extended lymph node dissection,1 underwent PD + portal vein reconstruction,1 underwent pancreatectomy,4 underwent resection of body and tail of pancreas and splenectomy.The size of the tumors ranged from 2.0 cm × 2.0 cm × 2.0 cm to 14.0 cm × 12.0 cm × 9.0 cm.Duodenal and biliary invasion was observed in 4 patients,superior mesenteric vein-portal vein invasion in 1 patient,splenic artery invasion in 1 patient.Neural invasion was observed in 8 patients,including 4 patients with lymph node metastasis.The results of immunohistochemical staining showed that 10 patients were with positive expression of cytokeratin 7,and 1 patient was with positive expression of vimentin.Nine patients were followed up,2 patients did not receive postoperative chemoradiotherapy,6 received chemotherapy with Gemicitabine and 1 received interventional treatment.The survival time of the 9 patients ranged from 8 to 20 months,and the median survival time was 12 months.Eight patients died of tumor recurrence and metastasis.Conclusions Pancreatic anaplastic carcinoma is a distinct rare variant of pancreatic ductal adenocarcinoma.Pancreatic anaplastic carcinoma has high malignancy,definite diagnosis depends on pathological examination,and the surgical plan should be made according to the result of imaging examination.Conventional radiotherapy and chemotherapy are ineffective for the treatment of this disease.

Objective To investigate the expression of slit diaphragm proteins of glomerular podocyte,such as NEPH1 and Nephrin in type Ⅴ lupus nephritis (V-LN). Methods Twenty-five patients with V-LN and 18 patients with idiopathic membranous nephritis (IMN) were enrolled into the study, and 5 normal renal samples were the normal control group. Twenty-four hours urine protein excretion, serum albumin, creatinine, triglyceride, total cholesterol, serum C3, C4, urine C3 and NAG were tested respectively.Glomerular lesions were measured by light microscopy. The expressions of NEPH1 and Nephrin were determined by indirect immunofluorescent staining. The statistical treatment was used t-test. Results Compared to the IMN group, the 24 hours urine protein excretion and the concentrations of serum albumin, creatinine, urine C3 were not significantly different while the triglyceride, total cholestorel, serum C3, C4 were significantly decrease in the V-LN group (P<0.05). Urine NAG was increased in the V-LN group (P<0.01). By indirect immuno-fluorescent histochemitry examination, the glomerular expressions of NEPH1 and Nephrin were significantly decreased in both V-LN and IMN. Compared with the IMN group, the decrease of NEPH1 and Nephrin expression was more remarkable in the V-LN group. Conclusion The expression changes of NEPH1 and Nephrin may play an important pathogenic role in proteinuria of Ⅴ lupus nephritis. Renal tubular epithelial cell damage may play a role in proteinuria of V-LN.

Objective To analyze the surgical treatment strategy and prognostic factors of pancreatic neuroendocrine tumor. Methods The clinical data of 30 patients who underwent pancreatic surgery in our department from April, 1999 to May, 2007 were retrospectively reviewed. According to the new WHO classification system, factors possibly influence the long term survival, such as tumor size, operation types, pathological types, were analyzed. Results There were 18 males and 12 females, with a mean age of 54 years old (28 ～ 78 years old). Five patients were lost in follow up. Among 25 patients who were completely followed, 20 cases were benign and other 5 were malignant, the median survival time of benign and malignant group was 74.8 months and 33.8 months, respectively (X2 = 8.90, P = 0.003). Overall 1, 2, 3, 4, 5-year survival rates were 100%, 100%, 82. 0%, 82. 0%, 65. 6%, respectively. Conclusions Pancreatic neuroendocrine tumor was a rare type of tumor of the pancreas with a good prognosis if surgically resected. The new classification method was effective in predicting the prognosis.

OBJECTIVETo study the chemical constituents of the red alga Acanthophora spicifera boergesen aiming at searching for bioactive leading compounds.

METHODCompounds were isolated by various chromatographic techniques including column chromatography over normal phase silica gel and Sephadex LH-20 gel and reverse phase HPLC as well as recrystallization. Their structures were determined by spectroscopic methods including IR, MS, 1D and 2D NMR techniques. MTT method was used for testing cytotoxicity of compounds against human cancer cell lines HCT-8, Bel-7402, BGC-823, A549 and HELA. Their inhibition against proliferation of dog vascular smooth muscle cells was also screened by MTT assay.

RESULTSix sterols were isolated from the ethanolic extract of the red alga Acanthophora spicifera. Their structures were identified as 6-hydroxycholest-4-ene-3-one (1), cholest-4-ene-3, 6-dione (2), cholest-5-ene-3 beta-ol (3), 5 alpha-cholestane-3, 6-dione (4), beta-sitosterol (5) and saringosterol (6).

CONCLUSIONCompounds 1-3 and 5 were obtained from this genus for the first time. Compounds 1, 2 and 4 showed moderate cytotoxic activity against human cancer cell lines.

Cell Line, Tumor ; Humans ; Inhibitory Concentration 50 ; Methanol ; chemistry ; Rhodophyta ; chemistry ; Sterols ; isolation & purification ; pharmacology

The aim of this study was to build a gene chip system with surface plasmon resonance (SPR) technique, for which Gamma-peptide nucleic acid (Gamma-PNA) functioned as a probe, in order to improve sensitivity and its specificity. With the use of self-assembled monolayer (SAM) technology, surface chemistry of two-dimensional structure was used. Gamma-PNA was designed according to the bioinformatics, and was plated on the SPR chip modified by SAM. Subsequently, relevant parameters of the experiment were ensured and optimized. The results showed that the performances of Gamma-PNA probe was little affected by the ion concentration of buffer, and it had a strong light signal in a stable state. As the ion concentration was 0, there were still good hybrid reactions; pH value had less influence upon Gamma-PNA probe, and acid environment of buffer could be better. Gamma-PNA probe combined with sensor technologies achieved made the probe with dispensable labels and real-time detection. It also improved the efficiency of the hybridization and the stability, providing the foundation for clinical application.
Nucleic Acid Hybridization ; Nucleic Acid Probes ; Oligonucleotide Array Sequence Analysis ; methods ; Peptide Nucleic Acids ; genetics ; Sensitivity and Specificity ; Surface Plasmon Resonance