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Objective To investigate the low density lipoprotein receptor (LDLR)gene and apolipoprotein (Apo) B gene mutation in a Chinese family with familial hypercholesterolemia(FH) and give the kindrids clinical check-ups. Methods After physical examination, the kindreds underwent ECG and ultrasound checks. Blood samples were tested for lipid profiles. The promoter and all eighteen exons of LDLR gene were investigated by using PCR and agarose gel electrophoresis in combination with DNA sequence analysis. The results were compared with the normal sequences in GenBank and FH database ( www. ucl. ac. uk/fh ) to find mutations. In addition, the apolipoprotein B100 gene for known mutations (R3500Q,R3531C,R3501W and R3480W)that cause familial defective ApoB100 (FDB)was also tested using the same method. Results A novel homozygous G ＞ A mutation at the 1581 bp of exon 10 was detected in the proband and his siblings. It caused a substitution of amimo acid Glu to Gly at codon 496. A novel heterozygous G ＞A mutation at the 1581 bp of exon 10 was detected in his parents. No mutations of R3500Q,R3531C,R3501W and R3480W of ApoB100 were observed. ECGs were normal. Atherosclerosis were found in all family members by ultrasound checks. Conclusions The homozygous G ＞ A mutation at the 1581 bp of exon 10 was first determined in our country. The change of amino acid Glu to Gly is responsible for FH of the family. The type of the gene mutation was not found in the FH database( www. ucl.ac. uk/ih). It's a new type of LDLR mutation.
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<p><b>BACKGROUND</b>To investigate the relationship between micrometastasis of bone marrow and neoadjuvant chemotherapy and prognosis in patients with stage III non-small cell lung cancer (NSCLC).</p><p><b>METHODS</b>Sixty-five patients with stage III NSCLC were randomly divided into group A (32 patients treated with preoperative neoadjuvant chemotherapy plus operation) and group B (33 patients treated with operation and postoperative chemotherapy as control group). Expression of CK19 mRNA and CEA mRNA was detected in bone marrow samples from the rib segments of all patients obtained from operation by RT-PCR. The relationship between survival duration and CK19 and CEA expression was analyzed.</p><p><b>RESULTS</b>The positive rates of CK19 mRNA expression were 18.8%(6/32) and 45.5%(15/33) in group A and B, respectively (P=0.033), and the positive rates of CEA were 25.0%(8/32) and 51.5% (17/33) in group A and B, respectively (P= 0.041). A significant positive correlation was observed between CK19 and CEA expression (r s=0.671,P < 0.001). The response rates of neoadjuvant chemotherapy were 0%(0/5) and 56.5%(13/23) in patients with CK19(+)/CEA(+) and CK19(-)/CEA(-), respectively (P=0.044), and the median survival duration were 11 and 27 months, respectively (P=0.000 6). Cox's model showed that the response to neoadjuvant chemotherapy and the expression of CK19 or CEA were significantly prognostic factors in group A.</p><p><b>CONCLUSIONS</b>Neoadjuvant chemotherapy can reduce the possibility of bone marrow micrometastasis in stage III NSCLC patients. Bone marrow micrometastasis may indicate a poorer prognosis for NSCLC.</p>
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<p><b>OBJECTIVE</b>To study the relation between the expression of transcription factor T-bet/GATA3 and Th1/Th2 type cytokines in peripheral blood mononuclear cells (PBMC) from lung cancer patients and their interference by the traditional Chinese herbal medicine.</p><p><b>METHODS</b>The gene expression of Th1/Th2 type cytokine IFN gamma, IL-2, IL-4, IL-6, IL-10, transcription factor T-bet/GATA3 and tumor tissue specific mRNA CEA, CK19 in PBMC from lung cancer patients were detected by reverse transcription-polymerase chain reaction RT-PCR. Meanwhile, the change of IFN gamma, IL-4, T-bet and GATA3 in PBMC before and after being cultured with the traditional Chinese herbal medicine-Astragulus and Tetramethylpyrazine was also observed.</p><p><b>RESULTS</b>Predominant expression of Th2 type cytokines was detected in 42 lung cancer patients. The positive rates of IL-4, IL-6, IL-10, IFN gamma and IL-2 were 27/42, 24/42, 31/42, 4/42 and 5/42, respectively. But, the positive rates of transcription factor T-bet and GATA3 were 16/42 and 34/42. Moreover, the expression intensity of T-bet was lower in the CEA and CK19 positive patients than the negative ones. On the contrary, the expression intensity of GATA3 was significantly higher in the same patients.</p><p><b>CONCLUSION</b>Predominant expression of Th2 type cytokines may be related to lower expression of T-bet or higher expression of GATA3. This condition can be interfered by the traditional Chinese herbal medicine-Astragulus and Tetramethylpyrazine.</p>
Subject(s)
Humans , Cytokines , Blood , DNA-Binding Proteins , Genetics , Drugs, Chinese Herbal , Pharmacology , GATA3 Transcription Factor , Gene Expression , Lung Neoplasms , Blood , Genetics , Medicine, Chinese Traditional , RNA, Messenger , T-Box Domain Proteins , Th1 Cells , Allergy and Immunology , Th2 Cells , Allergy and Immunology , Trans-Activators , Genetics , Transcription Factors , GeneticsABSTRACT
Objective To investigate the expression of FHITmRNA and WWOXmRNA in human breast cancer tissues and its relation to clinicopathological and other molecular parameters. Methods With reference to the expression of ?-actin,the expression of FHITmRNA and WWOXmRNA was determined by reverse (transcription)-polymerase chain reaction(RT-PCR) in 51 breast cancer and adjacent breast tissue, and (semi-quantitative) analysis of band densities was performed. The protein expression of estrogen receptor(ER), progesterone receptor (PR), Her-2 gene in the 51 breast cancer lesions was detected by (immunohistochemical) method. Results FHITmRNA and WWOXmRNA expression was significantly different in 54 breast cancer tissue compared to adjacent breast tissue (P0.05); of FHITmRNA and WWOX mRNA was related to axillary lymph node metastasis (P
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AIM: In this paper, we studied the efficiencies and the mechanisms of a new Chinese herb Qcimum basilicum polysaccharide (BP) on PG cell metastasis in vitro. METHODS: The number of tumor cells going through matrigel was assayed and used to represent the ability of the invasion and migration of PG cells. Using Scrape-loading and dye transfer (SLDT) technique, the efficiencies of BP on recovering PG cell gap junction -mediated intercellular communication (GJIC) was measured. The expressions of c-myc, nm23-H1 and Tiam-1 genes mRNA in PG cells treated with BP were determined by RT-PCR. RESULTS: Compared with the control, the action of invasion and migration of PG cells were decreased after treated with BP (P
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Objective:To study the role of IL-2,IL-4,IL-6,IL-10,IL-13 and IFN? in pathogenesis of human tumor.Methods:The gene expression of 6 cytokines in human tumor cell lines and specimens were detected by RT-PCR or RNA dot hybridization.Results: There were predominant expression of Th2 type cytokines in 183 different human tumor cell lines and specimens.The positive rate of IL-4,IL-6,IL-10,IL-13 .IFN? and IL-2 in 183 tumor cell lines and specimens were 65%,70.5%,83.6%,61.7%, 12.6% and 22.9%,respectively.Conclusion: Predominant expression of Th2 type cytokines play an important role in the pathogenesis and development of tumor, may be related to the evasion of tumor cells from immune surveillance.
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Objective:To observe the expression of Th1 versus Th2 type cytokines in primary hepatic cancer(PHC)and its adjacent liver tissues.Methods:The gene expression of Th1/Th2 cytokines was detected by RT-PCR using IFN-?and IL-2 as Th1 type cytokine genes,IL-4 and IL-10 as Th2 type cytokine genes.Results:Thl type cytokines were expressed in 7 and 9 cases,while Th0 type cytokines in 4 and 2 among 11 PHC and their adjacent liver tissues,respectively.Conclusion:Th1 type cytokines are expressed predominantly in primary hepatic cancer and its adjacent liver tissue.
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Objective: To provide an efficeut protocol for constructing recombinant adenovirus, an in vitro ligation was used instead of homologous recombination. Methods: Gene BMP-2 was ligated into pShuttle 2 vector ( pShuttle 2-BMP-2 ) and then fragment containing BMP-2 gene and promoter pcmvie excised by PI-SCe Ⅰ and Ⅰ-Ceu Ⅰ endonuclease. The fragment was further combined with adenovirus vector (Adeno-X-BMP-2) , which was finally linearized with Pac Ⅰ and trans-fered to HEK293 to package adenovirus particles. Results: Both PCR assay and restiction analysis showed that the recombined rectors pShuttle2-BMP-2 and Adeno-X-BMP-2 contains the target BMP-2 gene. THe packaged adenovirus was also i-dentified by PCR assay with specific primers for BMP-2. Conclusions: The BMP-2 incorporated recombinant adenovirus was obtained and this laid a foundation for further study on BMP-2 mediated gene therapy. The in vitro ligation method de-scinbed here for constructing recombined adenovirus was more efficient than traditional homologous recombination.
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Objective To study the expression of BRMS1mRNA in human breast cancer tissues and their significance.Methods The expression of BRMS1mRNA was detected by reverse transcription-polymerase chain reaction(RT-PCR) in 71 breast cancer tissues and adjacent breast tissues,12 patients with benign breast tumors and 12 patients with normal breast tissue,and semi-quantitative analysis of band densities was also performed.Results The expression of BRMS1mRNA in 71 patients with breast cancer and adjacent breast tissue was 0.378?0.046 and 0.918?0.044,respectively;the expression of BRMS1mRNA in 12 patients with benign breast tumors and 12 patients with normal breast tissue was 0.908?0.047 and 0.934?0.028 respectively.BRMS1mRNA expression was significantly lower in breast cancer tissue compared to adjacent breast tissue,benign breast tumors and normal breast tissue(P0.05),but was related to axillary lymph node metastasis and clinical stage(P