Association of chemokine and chemokine receptor expression with the invasion and metastasis of lung carcinoma.

The chemokine system has been reported to be utilized and manipulated by tumor cells in order to promote local tumor growth and distant dissemination. The present study aimed to investigate the expression of three chemokine ligand-receptor axes in lung carcinoma tissues. Tumor and healthy normal tissue samples were obtained from 120 lung carcinoma patients following surgical resection.

KRAS and BRAF gene mutations and DNA mismatch repair status in Chinese colorectal carcinoma patients.

Aim: To investigate gene mutations and DNA mismatch repair (MMR) protein abnormality in Chinese colorectal carcinoma (CRC) patients and their correlations with clinicopathologic features.

Methods: Clinical and pathological information for 535 patients including 538 tumors was reviewed and recorded. Mutation analyses for exon 2 of KRAS gene and exon 15 of BRAF gene were performed by Sanger sequencing except that in 9 tumors amplification refractory mutation system PCR was used.

Mast cells or not? - CD117 positive cells in esophageal leiomyoma.

The presence of CD117 positive cells in esophageal leiomyoma may lead to a misdiagnosis of GIST. We reviewed 46 esophageal tumors which were smooth muscle tumors or GIST. Based on morphology, immunohistochemistry and mutation analysis, there were 44 (95.

[Anaplastic lymphoma kinase fusion gene expression, clinical pathological characteristics and prognosis in 95 Chinese patients with non-small cell lung cancer].

Objective: To examine the prevalence of anaplastic lymphoma kinase (ALK) fusion gene in Chinese patients with non-small cell lung cancer (NSCLC).

Methods: In this study, 95 patients with NSCLC and corresponding clinical information and formalin-fixed paraffin-embedded (FFPE) tissue blocks were included. Hematoxylin & eosin (HE) staining, conventional ALK immunochemistry (IHC) staining and intercalated antibody-enhanced polymer (iAEP) IHC staining, and dual-color split fluorescence in situ hybridization (FISH) for ALK fusion gene were performed.

Screening for EGFR and KRAS mutations in non-small cell lung carcinomas using DNA extraction by hydrothermal pressure coupled with PCR-based direct sequencing.

EGFR and KRAS mutations correlate with response to tyrosine kinase inhibitors in patients with non-small cell lung carcinoma (NSCLC). We reported a hydrothermal pressure method of simultaneous deparaffinization and lysis of formalin-fixed paraffin embedded (FFPE) tissue followed by conventional chaotropic salt column purification to obtain high quality DNA for mutation analysis using PCR-base direct sequencing. This study assessed the feasibility of using this method to screen for exons 18-21 of EGFR and exon 2 of KRAS gene mutations in surgical resection and core needle biopsy specimens from 251 NSCLC patients.

[Application of combined telomerase activity analysis and immunocytochemistry in cytopathologic diagnosis].

Objective: To evaluate the application of traditional cytomorphology, telomerase activity analysis and immunocytochemistry in cytopathologic diagnosis of pleural effusion and bronchoalveolar lavage samples.

Methods: A total of 123 agar-paraffin double-embedded pleural effusion and bronchoalveolar lavage fluid samples were enrolled into study. The cytomorphologic features were reviewed and correlated with immunocytochemical findings and telomerase activity.

Quantification of alternative splicing variants of human telomerase reverse transcriptase and correlations with telomerase activity in lung cancer.

Telomerase plays important roles in the development and progression of malignant tumors, and its activity is primarily determined by transcriptional regulation of human telomerase reverse transcriptase (hTERT). Several mRNA alternative splicing variants (ASVs) for hTERT have been identified, but it remains unclear whether telomerase activity is directly associated with hTERT splicing transcripts. In this study, we developed novel real-time PCR protocols using molecular beacons and applied to lung carcinoma cell lines and cancerous tissues for quantification of telomerase activity and three essential hTERT deletion transcripts respectively.

Detection of telomerase activity in cultured cells and tumor tissue of lung carcinoma by modified telomeric repeat amplification protocol.

Telomerase activity is found in various cell types including stem cells, neoplastic cells, and immortalized cells, suggesting a close association with their proliferation capacity. The telomeric repeat amplification protocol (TRAP) has been traditionally used to detect semi-quantitatively the telomerase activity by polyacrylamide gel electrophoresis (PAGE), which is difficult to apply for large scale analysis because of laborious post-PCR manipulation and potential carryover contamination. In the present study, a specific reverse primer was designed and the TRAP protocol was adapted to either PAGE or real-time PCR assay.

EGFR mutations are more frequent in well-differentiated than in poor-differentiated lung adenocarcinomas.

Somatic mutations in epidermal growth factor receptor (EGFR) tyrosine kinase domain, particularly deletions in exon 19 and point mutation in exon 21, are associated with clinical outcome in patients with lung adenocarcinoma, suggesting that EGFR mutation would have an important role in clinical decision making. DNA was extracted from the excised specimens of 60 lung adenocarcinoma patients with phenol-chloroform and ethanol precipitation. Exon 19 and 21 were amplified by PCR, and direct sequenced from both sense and antisense directions.

[A clinicopathologic study on complications of orthotopic liver transplantation in 54 patients with chronic hepatitis B virus infection].

Objective: To study the complication incidence of 54 patients with chronic HBV infection following their orthotopic liver transplantation (OLT), and factors associated with HBV recurrence and hepatocellular carcinoma (HCC) recurrence or metastasis post-OLT.

Methods: The light-microscopic appearance of hepatic allograft biopsies in 54 patients with chronic HBV infection following OLT was examined. The related clinical data were analyzed.

[P2Y purinergic receptor activated PI-3K/Akt signaling pathway in regulation of growth and invasion of prostatic cancer].

Objective: To investigate P2Y purinergic receptor activated PI-3K/Akt signaling pathway in the regulation of growth and invasion of prostate cancer in vitro.

Methods: Western blot was used to detect phosphorylation of Akt (a downstream target molecule of PI-3K) by P2Y receptor agonist in 1E8 cells (a highly metastatic subclone derived from PC-3 prostatic cancer cell line). Cell counts, flow cytometry, Matrigel invasion assay, wound healing assay and gelatin zymography were used to detect changes of biological behaviors of 1E8 cells after P2Y receptor activation.

[Status and clinicopathologic implication of epidermal growth factor receptor mutation in non-small cell carcinoma of lung].

Objective: To investigate mutations of epidermal growth factor receptor (EGFR) exon 19 and 21 in non-small cell lung carcinoma and to explore their clinicopathological correlations.

Method: DNA was extracted from the excised tumor specimens of 66 non-small cell lung carcinoma patients by traditional phenol-chloroform and ethanol precipitation. Exons 19 and 21 were amplified by polymerase chain reaction (PCR), followed by direct sequencing in both sense and antisense directions.

[Detection of Mycobacterium tuberculosis DNA in sarcoidosis samples using real-time fluorescence polymerase chain reaction].

Objective: To study the role of Mycobacterium tuberculosis in the pathogenesis of sarcoidosis.

Methods: Archival material of 22 patients with a histologic diagnosis of sarcoidosis were retrieved. Real-time fluorescent polymerase chain reaction (PCR) was used to detect DNA fragments of the complex-specific insertion sequence IS6110 of Mycobacterium tuberculosis in formalin-fixed and paraffin-embedded biopsy samples.

[Status and clinical implication of c-kit and PDGFRA mutations in 165 cases of gastrointestinal stromal tumor (GIST)].

Objective: To investigate the status of c-kit and PDGFRA mutations of GIST in a the large sample of Chinese patients.

Method: One hundred and sixty-five cases were evaluated for the presence of c-kit and PDGFRA mutations. Exon 9, 11, 13, 17 of c-kit and exon 12, 18 of PDGFRA were analyzed by PCR amplification and direct sequencing.

[PTCH gene mutations in odontogenic keratocysts].

Objective: To investigate the frequency, type and distribution of PTCH mutations in odontogenic keratocysts (OKC) and to analyze the molecular pathological relationship between sporadic OKC and OKC associated with nevoid basal cell carcinoma syndrome (NBCCS).

Methods: Genomic DNA was extracted from 8 cases of OKC lesions (4 sporadic OKCs and 4 NBCCS-related OKCs). PTCH gene mutations were detected by PCR-direct sequencing.

[c-kit and PDGFRA mutations in 60 cases of gastrointestinal stromal tumors (GISTs)].

Objective: To explore the status of activating mutations of c-kit and PDGFRA in GIST of Chinese patients.

Methods: Sixty GISTs, confirmed by immunoreactivity of CD117, CD34, SMA, S-100 and Desmin, were evaluated for the presence of c-kit exons 9, 11, 13 and 17 mutations, and PDGFRA exons 12 and 18 mutations. The PCR products were sequenced directly for mutations, using DNA extracted from paraffin-embedded tissue.

[Correlation of expression of connexin to growth and progression of cervical carcinoma in situ].

Background & Objective: Gap junction intercellular communication (GJIC), mediated by connexin (CX), is the unique type of intercellular communication in carcinoma in situ (CIS). Changes in expression of CXs and function of GJIC may play roles in carcinogenesis of cervical cancer and progression of CIS. This study was to investigate the expression of CXs in normal epithelium (NE), hyperplasia, CIS, and invasive carcinoma (IC) of cervix, to explore correlation of expression of CXs to pathogenesis and progression of cervical CIS.

[Activation of HIF-1 by bFGF in breast cancer: role of PI-3K and MEK1/ERK pathways].

Objective: To explore the mechanism of basic fibroblast growth factor (bFGF)-mediated hypoxia inducible factor (HIF-1) activation and the down-stream signaling pathways involved.

Methods: Human breast cancer cells of the line T47D were cultured and lysed to extract the total protein in the supernatant. The amounts of extracellular signal kinase 1/2 (ERK1/2), Akt, p38, and beta-tubulin were measured.

[Diagnosis of lymph node micrometastases in human lung carcinoma: determination of p53 gene mutations by polymerase chain reaction-temperature gradient gel electrophoresis (PCR-TGGE)].

Background & Objective: Metastasis is one of the most important factors in determining the prognosis of lung cancer patients. However, it is difficult to determine single cancer cell in lymph nodes by routine methods. This study was designed to investigate a sensitive method for detecting lymph node micrometastases in human lung carcinomas.

[A novel Krit-1 mutation in Han family with cerebral cavernous malformation].

Objective: To detect the mutations of Krit-1 gene that cause familial cerebral cavernous malformation (CCM) in the Han ethnic origin.

Methods: The subjects were hospitalized in the Department of Neurosurgery, Tiantan Hospital affiliated to Capital University of Medical Sciences. Two families (A and B) and 8 apparently sporadic individuals affected with CCM were screened for mutations of Krit-1 gene.

[Gene detection of severe acute respiratory syndrome-related coronavirus].

Objective: To develop a newly real-time RT-polymerase chain reaction assay for severe acute respiratory syndrome (SARS) related coronavirus in human whole blood.

Methods: A pair of primers and a probe (molecular beacon) had been designed that were specific for the recognition of a highly conservative region between 15 301 and 15 480 of the SARS-related coronavirus polymerase gene sequences obtained from GenBank (G130027616).

Results: In the real-time RT-PCR assay, the extent of SARS related coronavirus amplification was measured in terms of the increase in fluorescence during the amplification process.

[Down-regulation of metastatic phenotype in human melanoma cells by controlled expression of anti-sense matrix metalloproteinase 9].

Objective: To investigate the correlation between matrix metalloproteinase 9 (MMP-9) expression and tumor invasion and metastasis as well, and to explore the potential application of controlled expression of target gene in tumor gene therapy.

Methods: One self-contained tetracycline-regulated retroviral vector containing anti-sense cDNA of MMP-9 was constructed and transfected into a metastatic human melanoma cell line WM451 which expressed a high level of MMP-9. Techniques such as growth rate measurment, MTT assay, (3)H-thymidine incorporation, colony forming ability in soft agar, invasion assay in Boyden chamber, as well as zymography and Western blot were applied to analyze the expression of MMPs and behaviors of tumor cells in vitro before and after gene transfection.

[Effects of two variants of ING1 expression on tumor cell growth regulation].

Objective: To study effects of alternative transcripts of ING1 transfection on human cancer cell lines.

Methods: p47/ING1A and p33/ING1B expression vehicles were constructed and introduced into a human breast cancer cell line MCF-7 and a human lung cancer cell line PAa, both expressing wild-type p53 protein. Growth characteristics of the transfectants and potentially related genes were analyzed.