Objective Insulin-like growth factor-1 receptor (IGF-1R),similar to insulin receptor,is one of the families of re- ceptor tyrosine kinases.,which has been found to be overexpressed in a variety of cancer.It is the main proliferation and survival sig- nal molecule in cancer cell and plays an important role in cancer growth and progress.Blocking signal transduction of IGF-1R by vari- ous strategies can suppress tumor growth and induce regression of established tumor.This study is to construct the siRNA expression vector targeting IGF-1R and to evaluate its ability to induce cell apoptosis in human lung cancer cells.Methods Two siRNA expres- sion vector,pENTR/U6-shRNA-1 and pENTR/U6-shRNA-2 targeting IGF-1R,were constructed using pENTR/U6 vector,and a vector targeting hieiferase gene,pENTR/U6-shRNA-Iuc,was constructed as control.After vectors were transfected into A549 for 48h, knockdown of IGF-1R mRNA and protein and Akt phosphorylation were accessed,and DNA ladder and flow cytometry were used for cell apoptosis.Results siRNA expression vectors targeting IGF-1R were successfully constructed,which was confirmed by PCR and DNA sequencing,pENTR/U6-shRNA-1 and pENTR/U6-shRNA-2 demonstrated the expression were (22.1?2.5) % and (80.1? 3.9) % in IGF-1R mRNA level,(15.2?3.1)% and (47.1?4.1)% in protein level,respectively,compared with pENTR/U6- shRNA-luc.Suppression of IGF-1R by pENTR/U6-shRNA-1 blunted Akt phosphorylation,increased cell apoptosis induced by 3% ethanol,and retained 77.5 % A549 cells in the G0/G1 phase.Conclusion siRNA expression vector targeting IGF-1R can effectively suppress the expression of IGF-1R expression in A549.This study suggests that DNA vector-based RNAi has the potential to be effec- tive and practical cancer gene therapy strategy.

Objective Overexpressions of epidermal growth factor(EGF)and EGF receptor have been associ- ated with progression and invasive phenotype of pancreatic cancer.However,the underlying molecular mechanism by which EGF worked in pancreatic cancer cells has not been completely understood.In this study,effect of EGF on the invasion and metastasis of pancreatic cancer cells and its regulatory mechanism were investigated.Methods The effects of EGF on the proliferation,adhesion and invasion of pancreatic cancer cells were detected by WST-1 prolif- eration assay,adhesion assay and invasive assay,respectively.The activity and expression of MMP-2 and MMP-9 were examined by zymography,Western blot and RT-PCR,respectively.The activity of NF-?B was examined by EMSA.Results EGF could significantly promote the invasiveness of pancreatic cancer cells but did not affect cell proliferation or adhesion.The expressions of NF-?B and MMP-9 were significantly increased by EGF,but EGF did not affect the activity and expression of MMP-2.Furthermore,EGF stimulated the NF-?B binding activity.Pre- treatment with NF-?B inhibitors,pyrrolidine dithiocarbamate(PDTC),could significantly inhibit the activity of NF-?B induced by EGF.Meanwhile,the EGF-induced expression and activity of MMP-9,as well as cell invasiveness were also inhibited by NF-?B inhibitor.Conclusion EGF could increase the expression and promote the invasiveness of MMP-9 via the activation of NF-?B in pancreatic cancer cells,which implies that NF-?B inhibitant,such as PDTC,may diminish the invasiveness of pancreatic cancer cells.

OBJECTIVETo investigate the relationship between (GT)n polymorphism and esophageal cancer by analyzing the connection between microsatellite polymorphisms in the promoter of heme oxygenase-1 and the clinicopathological characteristics of esophageal squamous cell carcinoma (ESCC) in Han chinese population.

METHODSThe (GT)n repeats in HO-1 gene in 83 male and 43 female hospital-based patients with ESCC (aged between 40 and 79 years with a mean of (61 ± 8) years) and 134 healthy control individuals were obtained by DNA sequencing. Polymorphisms of the (GT)n repeats were generally grouped into three classes based on allele frequencies: class S alleles (<25 repeats), class M alleles (25 to 29 repeats), and class L alleles (≥30 repeats). The correlation between susceptibility and clinicopathological characteristics of ESCC were analyzed by χ2 test. For in vitro experiments, the transient-transfection assay was performed to explore the correlation between different lengths of (GT)n repeats and promoter activity by assessing the promoter activities of HO-1 gene in cultured Ecal09 cells treated with H2O2 by analysis of cariance.

RESULTSHigher frequencies of L-allele (25. 8% vs. 14. 9%, χ2 = 9. 520, P = 0. 002), L-allele carrier (41. 3% vs. 27. 6%, χ2 = 5. 381 , P = 0. 020) were found in patients with ESCC. Furthermore, the lymphatic metastasis rate (63. 5% vs. 41. 8%, χ = 5. 685, P = 0. 017) and the detection rate of poorly differentiated ESCC cell (53. 8% vs. 28. 4%, χ2 = 8. 335, P = 0. 004) was significantly higher in L-allele carriers compared to non-L-allele carriers. In transfection experiments, promoter activities of 5'-flanking regions of the HO-1 gene in Eca109 cells transfected with the recombinant gene carrying (GT)16 repeat after treatment with H2O2 increased (F = 23. 615,P = 0. 008). In H2O treated control group, compared to (GT)26 and (GT)36, the basal promoter activities of HO-1 gene carrying (GT)16 repeat increased (F =41. 376, P = 0. 003; F = 50. 761, P = 0. 002).

CONCLUSIONThe long (GT)n repeats of HO-1 gene promoter can increase the susceptibility of esophageal squamous cell carcinoma and the risk of lymphatic metastasis.

Alleles ; Asian Continental Ancestry Group ; Carcinoma, Squamous Cell ; etiology ; pathology ; Esophageal Neoplasms ; etiology ; pathology ; Female ; Gene Frequency ; Heme Oxygenase-1 ; genetics ; Humans ; Hydrogen Peroxide ; Lymphatic Metastasis ; Male ; Microsatellite Repeats ; Polymorphism, Genetic ; Promoter Regions, Genetic ; Risk Factors ; Transfection

Peptide cyclization, a pivotal approach to modifying linear precursors of proteins and pepticles, has been used to enhance their biological activities and serum stabilities. Recently, sortase A (SrtA) from Staphyloccus aureus becomes a promising new technology for efficiently incorporating site specific modifications into proteins, conjugating the cell surface and cyclizing the linear peptides. In this study, we constructed two recombinant expression systems, one with chitin binding domain and the other with six-histidine tag and chitin binding domain on the N-terminal of SrtA, separately. The results of enzymatic kinetics indicate that the two recombinant tags do not impair the transpeptidase activity of SrtA compared with the standard reaction reported under the same reaction condition. The two synthesized peptides with N-ternimal three glycines and C-terminal penta-amino acid motif, LPETG, were cyclized using immobilized and recycled SrtA. The SrtA-based cyclization promises to represent a simple method for easy and efficient enzymatic synthesis of large cyclic peptides.
Aminoacyltransferases ; metabolism ; Bacterial Proteins ; metabolism ; Cyclization ; Cysteine Endopeptidases ; metabolism ; Enzymes, Immobilized ; metabolism ; Kinetics ; Peptides ; metabolism ; Peptides, Cyclic ; biosynthesis ; Staphylococcus aureus ; enzymology

Intrahepatic cholangiocarcinoma (ICC) is the second most common primary liver cancer with a very poor prognosis.In order to guide better clinical management of ICC patients,the American Joint Committee on Cancer (AJCC) cancer staging manual (7th edition) have established a unique TNM staging scheme for separating ICC from hepatocellular carcinoma (HCC) for the first time,and reflected a difference between risk factor of ICC and HCC.This TNM staging system for ICC has been most recently updated by the AJCC cancer staging manual (8th edition),in which T staging has been redefined without gross features,and lymph node metastasis (N1) in N staging has been grouped as stage Ⅲ B,but not stage Ⅳ as required by the 7th edition of AJCC cancer staging manual.In addition,region lymphatic and distant metastases have been clearly redefined by the AJCC cancer staging manual (8th edition) that also requires recovering at least 6 lymph nodes for the N staging scheme.The apparent advantages of the AJCC cancer staging manual (8th edition) for ICC pathologic staging may better stratify the prognosis of ICC patients and provide an improved guidance in clinical practice.

BackgroundThe immunotherapy for retinoblastoma(RB) is gradually concerned recent year.To seek relative immune-associated antigen is a basis of immunotherapy.NY-ESO-1 and NY-SAR-35 are two kinds of genes of cancer testis antigen( CTA ).To understand their expressions in RB tissue can offer index for relative study.ObjectiveThis study was to investigate the expressions of two CTA NY-ESO-1 and NY-SAR-35 in RB and explore the possibility of them as potentially promising targets for antigen-specific immunotherapy of RB.Methods The samples were obtained from 15 RB eyes,12 non-tumor retinopathy eyes and 22 normal eyes with other benign eye diseases.Reverse transcription polymerase chain reaction(RT-PCR) was used to detect the expressions of NY-ESO-1 mRNA and NY-SAR-35 mRNA in the samples.Genes of positive PCR results were sequenced randomly.The relevance of the expression of the two cancer-testis antigen genes with the clinical characteristics such as tumor stage,tumor size and clinical stage were analyzed.This study was approved by Ethic Committee of Guangxi Medical University.Written informed consent was obtained from each patient before operation. Results NY-ESO-1 mRNA was positively expressed in 6 RB samples and NY-SAR-35 mRNA was expressed in 9 RB samples.In the non-tumor retinopathy samples and normal eye tissues,NY-ESO-1 mRNA and NY-SAR-35 mRNA were absent.No significant relevances were found between the expressions of the NY-ESO-1 mRNA and NY-SAR-35 mRNA with clinical characteristics such as age ( P =0.426,0.822 ),gender ( P =0.180,0.464 ),pathological classification ( P =0.744,0.582 ),tumor size ( P =0.760,0.790),and clinical stage ( P =0.868,0.707 ).Conclusions NY-ESO-1 and NY-SAR-35 have high expressing frequencies in RB tissue and their expressions in RB have specificity.These results offer a clue for the identification of targets antigen of RB.

Along with the development of Chinese medicine and pharmacy (CMP), Chinese materia medica (CMM) has been used more and more widely at home and abroad. But we have to confront worsening problems such as lack of safety evidence, immature self -formulation technologies, lack of knowledge about their toxicities, and public misunderstanding, especially for patients with cardiovascular disease (CVD). Therefore, we cardiovascular physicians are requested improve knowledge for CMM preparations and their effects and side effects, supervise and identify the interactions between CMM and Westem medicine. Meanwhile, the researchers are also requested to assess the safety and efficacy of CMM through rigorous experimental designs, further improve the quality, safety, and efficacy of CMM, strictly formulate the specification of CMM products, guide the rational use of CMM by clinicians and the general public.
Cardiovascular Diseases ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Humans ; Phytotherapy ; adverse effects ; Safety

OBJECTIVETo clarify the clinicopathologic significance of the expression of the Bcl-2 protein (pBcl-2) and the Bax protein (pBax), and their clinical implications in Chinese and Japanese patients with human invasive ductal carcinomas (IDCs) of the pancreas.

METHODSThe study included 59 Chinese and 65 Japanese patients with IDCs of the pancreas. pBcl-2 and pBax expression were immuno-stained with streptavidin-biotin (SAB) method.

RESULTSpBcl-2 (+) was seen in 35.6% of Chinese and in 23.1% of Japanese patients. pBax (+) was seen in 49.2% of Chinese and 64.7% of Japanese patients. A comparison between them showed that there were significant differences in the male patients, in the patients with the moderately differentiated cancer, and in the elderly patients (chi squared = 4.447, P = 0.035; chi squared = 4.114, P = 0.043; chi squared = 6.657, P = 0.010 respective). In both Chinese and Japanese patients, those with pBcl-2 positive expression had a significantly higher survival rate than those with negative one (chi squared = 9.99, P = 0.0016; chi squared = 7.63, P = 0.0058). The group with pBax positive expression had a significantly higher survival rate in Japanese patients (chi squared = 9.37, P = 0.0022). Japanese patients whose tumors exhibited pBcl-2 and pBax positive immunostaining survived significantly longer than Chinese patients did (chi squared = 4.48, P = 0.0342; chi squared = 5.23, P = 0.023).

CONCLUSIONSThe expressions of both pBcl-2 and pBax are high found in Chinese and Japanese patients. The pBcl-2 positive expression implies a better prognosis in both Chinese and Japanese patients with IDCs of the pancreas. The effect of pBax expression on prognosis is different between Chinese and Japanese patients.

Adult ; Aged ; Carcinoma, Pancreatic Ductal ; metabolism ; China ; Female ; Humans ; Immunohistochemistry ; Japan ; Male ; Middle Aged ; Pancreatic Neoplasms ; ethnology ; metabolism ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; bcl-2-Associated X Protein ; biosynthesis

OBJECTIVETo evaluate the method and clinical value of endoscopic surgery by comparing endoscopic resection and vaporization for superficial bladder tumor.

METHODS396 patients with superficial bladder papillary transitional cell carcinoma were treated by endoscopic therapy. 180 patients (Group A) were treated by transurethral resection of bladder tumor (TURBT) and 216 (Group B) by transurethral vaporization of bladder tumor (TVBT). Periodic postoperative intra-vascular instillation of chemotherapy was given to both groups. Operating time, amount of bleeding during operation, complications and recurrence rate were compared.

RESULTSIn group B, the amount of bleeding and complications during operation were lower than those in group A, but TVBT rated better by clearer view and simplicity in maneuver. The operating time, recurrence rate in group B were similar to those in group A.

CONCLUSIONTransurethral vaporization of bladder cancer, with simplicity in maneuver, less bleeding and fewer complications, rates better in effectiveness and clinical value than resection.

Adult ; Aged ; Aged, 80 and over ; Cystoscopy ; methods ; Electrosurgery ; methods ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Time Factors ; Urinary Bladder Neoplasms ; surgery

Three cyclotides were isolated from the whole plant of Viola yedoensis in this study. The two, vary peptide E and cycloviolacin Y5, were previously reported, and a novel cycloviolacin VY1 was characterized according to the interpretation of MS/MS fragmentation of peptides which were produced from the reduced and alkylated parent peptide with the digestion of Endo Lys-C, trypsin and chymotrypsin, separately. The stability of remarkable resistance to proteolytic degradation by trypsin and chymotrypsin, and that of thermal denaturation was confirmed again. Besides, the IC50 value of cycloviolacin VY1 against influenza A H1N1 virus was (2.27 +/- 0.20) microg x mL(-1). It is the first cyclotide reported with anti-influenza A H1N1 virus activity in vitro assay.
Antiviral Agents ; isolation & purification ; pharmacology ; Cyclotides ; pharmacology ; Influenza A Virus, H1N1 Subtype ; drug effects ; Tandem Mass Spectrometry ; Viola ; chemistry