Objective To observe the EGFR nuclear translocation in cervical carcinoma cell lines after irradiation and its possible role in radiation tolerance.Methods Western blotting was used to detect the nuclear EGFR and cytoplastic EGFR after irradiation.The effect of Cetuximab on expression of nuclear EGFR and survival fractions were investigated.Results After irradiation,compared with control group,the expression of nuclear EGFR protein increased in irradiated cervical carcinoma cell.Cetuximab inhibited the radiation-induced nuclear EGFR expression with decreased survival fractions.Conclusion Radiation could induce EGFR nuclear translocation in cervical carcinoma cell lines and nuclear EGFR might be correlated with radiation tolerance in Cervical carcinoma cell.

Celecoxib ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; metabolism ; Cyclooxygenase 2 Inhibitors ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; Kidney Neoplasms ; metabolism ; pathology ; Pyrazoles ; administration & dosage ; pharmacology ; RNA, Messenger ; metabolism ; Sulfonamides ; administration & dosage ; pharmacology ; Wilms Tumor ; metabolism ; pathology

The prevalence of thyroid cancer has shown an upward trend in China in recent years. Advances in thyroid ultrasound and fine needle puncture cytology have improved the accuracy of the preoperative diagnosis of thyroid cancer. Also,the application of endoscopy-assisted techniques and intraoperative nerve monitoring technology and the further understanding of thyroid lymph node metastasis have made the thyroid surgeries safer and less invasive. This article summarizes the recent advances in the surgical therapy of thyroid cancer.
Carcinoma, Papillary ; surgery ; Humans ; Iodine Radioisotopes ; therapeutic use ; Molecular Targeted Therapy ; Neoadjuvant Therapy ; Thyroid Neoplasms ; surgery ; therapy

OBJECTIVETo construct a vector expressing small interfering RNA (siRNA) against Rac1 gene and observe its effect on soft agar colony formation of SW480 cells in vitro.

METHODSOligos of 64 base pairs for hairpin RNA targeting Rac1 were chemically synthesized and annealed. The siRNA constructs for Rac1, produced by inserting the annealed oligos into the downstream of H1 promoter of linearized pSUPER, were confirmed by restriction digestion and DNA sequencing. The constructed Rac1-siRNA was transfected into SW480 cells and Western blotting was performed to assess the expression and interference efficiency of siRNAs against Rac1.The soft agar colony formation assay was used to study the effect of Rac1 gene silencing on SW480 cells.

RESULTSRestriction digestion and DNA sequencing showed that the siRNA targeting Rac1 gene was successfully constructed. The siRNA could effectively down-regulate the expression of Rac1 in SW480 cells. Soft agar colony formation assay showed that the colony number and diameter of SW480 cells was reduced after siRNA transfection.

CONCLUSIONA vector expressing hairpin RNA against Rac1 gene are successfully produced, which significantly reduces the colony numbers and size of SW480 cells in vitro, suggesting that Rac1 plays an important role in the growth of colorectal cancer in vitro.

Base Sequence ; Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; pathology ; Down-Regulation ; Humans ; Molecular Sequence Data ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; rac1 GTP-Binding Protein ; genetics

This study is to investigate inhibitory effects of lidamycin (LDM) on the proliferation of HERG K+ channel highly expressing cancer cells and its synergy with anticancer drugs. MTT assay was used to examine the inhibitory effects of lidamycin combined with various anticancer drugs on the proliferation of human lung cancer A549 cells, human colon cancer HT-29 cells and herg-stably-transfected A549 cells. Using the xenograft model of subcutaneously transplanted HT-29 in nude mice, inhibitory effect was appraised in vivo. The coefficient of drug interaction (CDI) was used to evaluate the synergistic effect of drug combination. LDM significantly inhibited the proliferation ofA549 cells and HT-29 cells with IC50 values of 2.14 and 4.64 ng mL(-1), respectively. The efficacy in HT-29 cells with high HERG potassium expression level is less potent than that in A549 cells with low expression level. In terms of IC50 values, LDM suppressed the growth of herg-stably-transfected A549 cells less potently than pCDNA3.1-stably-transfected A549 cells. There existed synergistic effects in the combinations of fluorouracil (5-FU) and LDM, doxorubicin (DOX) and LDM, or hydroxycamptothecine (HCPT) and LDM. CDI values of the combinations of 5-FU and LDM were more than 0.75. CDI values of LDM and DOX were more than 0.70, but some CDI values of LDM and HCPT were less than 0.70. As for the CDI values, synergistic effects of the combination of LDM and HCPT were the most potent of the three groups. There is no relationship between the inhibitory effect of the growth of cancer cells by 5-FU and HERG potassium expression level. HERG expression level negatively correlated with inhibitory effect on the proliferation of cancer cells by DOX. HERG expression levels and chemosensitivity were positively correlated for HCPT. In the model of subcutaneously xenograft transplanted HT-29 in vivo, LDM and/or HCPT effectively inhibited the growth of HT-29 in nude mice, and the optimum CDI of the combination of LDM and HCPT was less than 1. HERG expression level negatively correlates the chemosensitivity of cancer cells to LDM. There exist synergistic effects in vitro and in vivo in the combination of LDM and HCPT, which inhibitory effects of the proliferation of cancer cells positively modulated by HERG potassium expression level. HERG K+ channel may become a target of combined therapy for choosing anticancer drugs.
Aminoglycosides ; administration & dosage ; pharmacology ; Animals ; Antibiotics, Antineoplastic ; administration & dosage ; pharmacology ; Antineoplastic Agents, Phytogenic ; administration & dosage ; Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; Camptothecin ; administration & dosage ; analogs & derivatives ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Doxorubicin ; administration & dosage ; Drug Synergism ; ERG1 Potassium Channel ; Enediynes ; administration & dosage ; pharmacology ; Ether-A-Go-Go Potassium Channels ; metabolism ; Fluorouracil ; administration & dosage ; HT29 Cells ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Xenograft Model Antitumor Assays

OBJECTIVETo construct a rheumatoid arthritis-specific full-length fully human mammalian display antibody libraries.

METHODSPeripheral blood lymphocytes were isolated from patients with rheumatoid arthritis. The repertoires of kappa light chain (LCκ) and heavy chain variable region (VH) of the antibodies were amplified by RT-PCR. The amplified LCκ and VH genes were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO-10 strain to construct the rheumatoid arthritis-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length fully human antibody expressed on the surface of 293T cells were analyzed by flow cytometry.

RESULTSThe libraries of rheumatoid arthritis-specific antibody LCκ and heavy chain (IgG1) were constructed. The expression of full-length fully human antibody on the surface of 293T cells was confirmed by flow cytometry. With the rates of correct LCκ and heavy chain sequence insertion reaching 80% and 60%, respectively, as shown by DNA sequence analysis of the randomly selected clones, the libraries showed an expressible combinatory diversity of 6.13×10(10).

CONCLUSIONThe constructed libraries provide a useful platform for screening rheumatoid arthritis-specific antibodies.

Amino Acid Sequence ; Animals ; Antibodies ; genetics ; immunology ; Antibody Specificity ; Arthritis, Rheumatoid ; immunology ; Cell Surface Display Techniques ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; HEK293 Cells ; Humans ; Immunoglobulin G ; biosynthesis ; genetics ; Immunoglobulin Heavy Chains ; biosynthesis ; genetics ; Immunoglobulin kappa-Chains ; biosynthesis ; genetics ; Lymphocytes ; immunology ; metabolism ; Molecular Sequence Data ; Peptide Library ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Transfection

OBJECTIVETo test whether the tumor vessel density (TVD) from the enhanced spiral CT can preoperatively predict nodal status and distant metastasis of colorectal cancer.

METHODSForty cases of colorectal cancer patients who received surgical treatment were included in this study. The three dimensional tumor vessels were reconstructed by an enhanced CT 64-slice spiral CT and its AW4.4 image processing platform. The TVD was measured by the 1000 high-resolution color graphics pathological analysis system. The TVD level was compared between different tumor size, classification, and TNM stage. The postoperative pathological staging was taken as golden standard.

RESULTSThe sensitivity, specificity and accuracy for direct prediction of lymph node metastasis by the enhanced CT 64-slice spiral CT was 74.1%(20/27), 53.8%(7/13) and 67.5%(27/40) respectively. The TVD from the reconstructed three dimensional tumor vessels in the group with lymph node metastasis was significantly higher than that without metastasis(0.070±0.046 vs. 0.037±0.013, P<0.05). The TVD in the distant metastasis group was significantly higher than that without distant metastasis (0.130±0.032 vs. 0.049±0.030, P<0.01). No difference of TVD was found between different tumor size, invasion depth, and differentiation type.

CONCLUSIONTVD level from the reconstructed three dimensional tumor vessels can indicate lymph node and distant metastasis of colorectal cancer.

Colorectal Neoplasms ; Humans ; Lymph Nodes ; Lymphatic Metastasis ; Neoplasm Staging ; Tomography, Spiral Computed

OBJECTIVETo study clinical effects of modified radical mastectomy with preservation of major and minor pectoral muscles.

METHODSA retrospective analysis was carried out in 214 cases of breast cancer patients (including stage I 66 cases, stage II 141 cases and stage III 7 cases). Modified radical mastectomy with preservation of major and minor pectoral muscles was performed on all the patients.

RESULTSOut of 214 cases, 12 (5.6%) had subcutaneous fluid, 16 (7.4%) had skin flap margin necrosis. Upper limb lymphatic edema was found in 8 (3.7%) patients, and pectoral muscle contracture with dyspraxia of upper arm occurred in 11 (5.1%) cases. Three year survival rate was 82.3% and five year survival rate was 63.4%. For stage I patients, the five year survival rate attained to 79.6%, and stage II 56.3%.

CONCLUSIONSPreservation of lateral branch of pectoral nerve can avoid complication of pectoral muscle contracture with dyspraxia of upper arm. Early chemotherapy of postoperation prevents breast cancer occurrence and metastasis. Comprehensive treat approaches for operative wound avert subcutaneous fluid, and complex therapy improves long-term effect of breast cancer patients.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; drug therapy ; mortality ; surgery ; Female ; Follow-Up Studies ; Humans ; Mastectomy, Modified Radical ; methods ; Middle Aged ; Pectoralis Muscles ; surgery ; Retrospective Studies ; Survival Rate

OBJECTIVETo study the expression of connective tissue growth factor (CTGF) in the myocardial tissue of mice with viral myocarditis (VMC).

METHODBalb/c mice were infected with coxsackie virus B3 (CVB3) to establish VMC model. The mice were divided into control group (n = 50) and VMC group (n = 50). on days 4, 7, 14 and 21 after infection, heart specimens of 8 mice were randomly taken and examined after HE staining for myocardial necrosis and cellular infiltration. The area of positive Masson stained myocardium collagen fibers was measured, and collagen volume fraction (CVF) was measured. Then the level of serum creatine phosphokinase-MB (CKMB) was determined. The levels of CTGF and TGF-β₁ were detected by streptavidin peroxidase immunoperoxidase technique. Expression of CTGF and TGF-β₁ were detected with reverse transcription-polymerase chain reaction (RT-PCR). At the same time, the correlations were analyzed.

RESULT(1) The level of CKMB peaked on day 7, and decreased afterwards (455.45 ± 37.95, 606.95 ± 35.64, 573.62 ± 42.90, 308.60 ± 20.49, respectively, 4 - 21 d points), in which 4, 7, 14 d points, there was significant difference compared with control group (t = 6.144, 12.558, 11.182, respectively, P < 0.01). (2) CVF increased significantly on day 14 (8.22 ± 1.95, t = 4.486, P < 0.01) and day 21 (9.46 ± 1.87, t = 4.486, P < 0.01) in VMC group. (3) Measured by streptavidin peroxidase immunoperoxidase technique, the levels of CTGF (171.50 ± 10.25, 141.70 ± 10.863, 110.35 ± 11.051, 81.05 ± 10.190, respectively, 4 - 21 d points) and TGF-β₁ (184.90 ± 11.480, 150.25 ± 9.915, 103.50 ± 10.455, 84.15 ± 9.848, respectively, 4 - 21 d points) increased after day 4 in VMC (P < 0.01). (4) Measured by RT-PCR, the expression of CTGF mRNA and TGF-β₁ increased in VMC group, and the increase was enhanced with the disease development (P < 0.01). (5) The expression of CTGF and TGF-β₁ was positively linearly correlated (r = 0.987, P < 0.01), the expression of CTGF was negatively correlated with CVF (r = -0.901, P < 0.01), but the expression of CTGF was detected earlier than myocardial fibrosis.

CONCLUSIONThe increase of CTGF expression was associated with the severity of myocardial fibrosis in VMC. These results suggest that abnormal expression of CTGF may take part in the development of fibrosis in VMC.

Animals ; Connective Tissue Growth Factor ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Myocarditis ; metabolism ; pathology ; virology ; Myocardium ; metabolism ; pathology ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo investigate the nutritional risk in children with severe pneumonia using the Screening Tool for the Assessment of Malnutrition in Paediatrics (STAMP) and the association between nutritional risk and adverse clinical outcomes.

METHODSAccording to the STAMP score, 216 children with severe pneumonia were classified into high nutritional risk group (HR group; n=98), moderate nutritional risk group (MR group; n=65), and low nutritional risk group (LR group; n=53). Fasting blood samples were collected to measure the levels of insulin-like growth factor-1 (IGF-1), adiponectin, leptin, non-esterified fatty acid (NEFA), albumin, transferrin, prealbumin, and retinol binding protein (RBP). The adverse clinical outcomes were recorded.

RESULTSCompared with the MR and LR groups, the HR group had significantly lower serum levels of IGF-1, leptin, adiponectin, prealbumin, and RBP, as well as a significantly higher serum level of NEFA (P<0.05). Compared with the MR and LR groups, the HR group had a significantly higher proportion of children admitted to the intensive care unit and a significantly longer duration of mechanical ventilation (P<0.05). The HR group had a significantly longer mean hospital stay and a significantly higher incidence rate of complications compared with the LR and MR groups (P<0.05).

CONCLUSIONSNutritional risk screening has an important value in evaluating the clinical outcome of children with severe pneumonia, and children at a higher nutritional risk tend to have more adverse clinical outcomes.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Malnutrition ; etiology ; Pneumonia ; complications ; Risk