Adolescent ; Adult ; Child ; Female ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Lymphoproliferative Disorders ; diagnosis ; etiology ; therapy ; Male ; Middle Aged ; Retrospective Studies ; Transplantation, Homologous ; Young Adult

Objective To study the genotype of the thyroid hormone receptor ? (THRB) gene in a patient with thyroid hormone resistance syndrome. Methods The peripheral blood samples of the patient and his parents were collected, then DNA was isolated. PCR and direct sequencing techniques were performed to determine if there were mutations in their THRB gene. Results No mutation was found in exon 1-9. There was a point mutation in exon 10 of THRB which is a T to C transition in nucleotide 1658 resulting in the replacement of the normal Val (GTG) with an Ala (GCG) (V458A). The mutation was located in exon 10 of THRB gene and was a heterozygote. No mutation was found in THRB gene of his parents.Conclusion The gene diagnosis confirms that the patient has a mutation V458A located in the ligand binding area of THRB.

Objective To study whether carbon dioxide used to establish pneumoperitoneum has an influence on port-site and intraperitoneal implantation and metastasis of tumor cells. Methods R 15 hepatic cancer cells were injected into 30 Wistar rats’ peritoneal cavities 1 hour before operation, then the 30 Wistar rats were randomly divided into 3 groups: gasless group, helium group and carbon dioxide group. The suspension was exposed to the gas environment for 2 hours, all animals were killed after 28 days and the port-site and intraperitoneal implantation and metastasis of tumor cells were examined.Results On port-site, intestinal serous coat, mesentery, greater omentum and diaphragm, the weights of tumor cells, in carbon dioxide group were (326.7?230.3) mg, (626.2?215.9) mg, (476.2?204.8) mg,(2 536.5?906.7) mg and (384.5?149.9) mg respectively; in helium group were (235.6?107.3) mg, (414.2?148.4) mg, (261.8?92.6) mg, (1 633.4?247.3) mg and(220.0?57.9) mg; in gasless group were (145.0?42.4) mg, (221.5?108.2) mg, (212.5?109.6) mg, (797.5?335.9) mg and 113.0 mg.The weights of carbon dioxide group showed a significant increase, compared with helium group and gasless group (P 0.05). Conclusion The insufflation of carbon dioxide promotes intraperitoneal tumor implantation and growth compared with helium and gaslessness in a rat model.

Breast Neoplasms ; metabolism ; pathology ; surgery ; Carcinoma ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Fibroma ; metabolism ; pathology ; surgery ; Humans ; Keratins ; metabolism ; Middle Aged ; Vimentin ; metabolism

The purpose of this study is to identify the electrical activity of neuron, the existence of the transition from bursting pattern to spiking pattern and the ion mechanism of the bursting pattern. The intracellular electrical activity patterns of single neurons in the stomatogastric ganglion (STG) of crayfish were recorded when the extracellular calcium concentration ([Ca(2+)](o)) or calcium-dependent potassium channel blocker tetraethylammonium concentration ([TEA](o)) were changed, using intracellular recording method. These single neurons were also functionally isolated from the ganglion by application of atropine and picrotoxin which could block the inhibitory acetylcholine synapses and glutamatergic synapses respectively. When [Ca(2+)](o) was decreased by increasing EGTA, the membrane potential of the neuron was increased, and the electrical activity patterns were changed from the resting state with lower potential value (resting state of polarization) to the bursting pattern firstly, and then to the spiking pattern, at last to the resting state with higher potential value (resting state of depolarization). When [TEA](o) was increased, the membrane potential of the neuron was increased, and the electrical activity pattern was changed from the resting state with lower potential value (resting state of polarization) to the bursting pattern firstly, and then to the spiking pattern. The duration of the burst of the bursting pattern was increased. When [Ca(2+)](o) was increased or [TEA](o) was decreased, an inverse procedure of the electrical activity pattern was exhibited. On one hand, the results indicate that a single neuron can generate various electrical activity patterns corresponding to different physiological conditions, and the regularity of the transitions between different electrical activity patterns. On the other hand, the results identify that the initiation and termination of the burst in bursting pattern are determined by calcium-activated potassium conductance, which is adjusted by intracellular calcium concentration influenced by inward calcium current. It may be the ionic mechanism of generation of the bursting pattern in a single neuron.
Action Potentials ; physiology ; Animals ; Astacoidea ; physiology ; Calcium ; metabolism ; Calcium Channels ; metabolism ; Ganglia, Invertebrate ; physiology ; Neurons ; physiology ; Potassium Channels, Calcium-Activated ; metabolism

Aged ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Carcinoma, Large Cell ; metabolism ; pathology ; Diagnosis, Differential ; Histiocytic Sarcoma ; metabolism ; pathology ; surgery ; Hodgkin Disease ; metabolism ; pathology ; Humans ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Lymphoma, Large-Cell, Anaplastic ; metabolism ; pathology ; Male ; Melanoma ; metabolism ; pathology ; Receptors, Cell Surface ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; surgery

OBJECTIVETo study the effect of exogenous caveolin-1 on the growth of laryngeal squamous cell carcinoma and its mechanisms.

METHODSEukaryotic expression vectors containing human caveolin-1 gene were transfected into Hep-2 cell line, the positive clones with high expression of caveolin-1 were identified by fluorescence quantitative real time reverse transcriptase-polymerase chain reaction and Western Blot. Cell proliferation viability was tested by methyl thiazolyl tetrazolium assay, the protein expression of epidermal growth factor receptor (EGFR), P-EGFR, extracellular signal-regulated kinase 1, 2 (Erk1, 2), P-Erkl, 2 and caveolin-1 were detected by Western Blot. The combination of caveolin-1 and EGFR were studied by immunoprecipitation and Western Blot. The in vivo antitumor activity of caveolin-1 was tested in Hep-2 xenograft tumor models in athymic nude mice, and the protein expressions of P-EGFR, P-Erk1, 2 and caveolin-1 were examined by immunohistochemistry.

RESULTSThree of caveolin-1 stably transfected Hep-2 cell clones were established. MTT assay showed that the proliferation of caveolin-1 overexpression Hep-2 cell clones decreased significantly comparing with the control. Immunoprecipitation and western Blot showed that caveolin-1 and EGFR were combined in Hep-2 cell line. Comparing with the parental cell line and cells transfected with control vector, there were the lower phosphorylation of EGFR and Erk1, 2 in the caveolin-1 overexpression Hep-2 cell clones. In the xenograft tumor models in nude mice, caveolin-1 overexpression Hep-2 cell clones showed the slower growth, smaller tumor size and the lower phosphorylation of EGFR and Erk1, 2.

CONCLUSIONSOverexpression of caveolin-1 inhibits growth of Hep-2 cell line in vitro and in vivo, arresting EGFR-MAPK signal pathway may involve in its mechanism.

Animals ; Carcinoma, Squamous Cell ; metabolism ; Caveolin 1 ; pharmacology ; Cell Line, Tumor ; Humans ; Laryngeal Neoplasms ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Mitogen-Activated Protein Kinase Kinases ; metabolism ; Receptor, Epidermal Growth Factor ; metabolism ; Signal Transduction ; Transfection

OBJECTIVETo investigate the effects of caveolin-1 on the biologic behavior of laryngeal squamous cell carcinoma HEp2 cell line in vitro.

METHODSEukaryotic expression vector of human caveolin-1 gene was constructed and transfected into HEp2 cells by Lipofectamine. The clones stably overexpressing caveolin-1 were identified by real-time PCR and Western blotting. Cell proliferation viability was tested by MTT assay. Anchorage-independent growth was determined by assaying colony formation in soft agar. Flow cytometry was used to assess the cell cycle and apoptosis. The relative phosphorylation level of EGFR and ERK1/2 were detected by Western blotting. Localization of caveolin-1 and EGFR were studied by laser confocal laser scanning microscopy.

RESULTSThe expression vector of caveolin-1 was constructed and three clones stably overexpressing caveolin-1 were obtained. Comparing with the parental HEp2 cells, the transfected cells exhibited a slower growth rate and formed fewer colonies in soft agar. The results of FACS analysis revealed that overexpression of caveolin-1 resulted in the cell cycle arrest at G0/G1 phase and increased the apoptotic cell fraction. EGFR was found to colocalize with caveolin-1 in transfected cells by confocal laser scanning microscopy and Western blotting results showed that overexpression of caveolin-1 reduced the phosphorylation of EGFR and Erkl/2.

CONCLUSIONOverexpression of caveolin-1 suppresses the growth of HEp2 cells and induces apoptosis and inhibition of EGFR-MAPK signaling pathway may be involved in its mechanism.

Apoptosis ; Blotting, Western ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Caveolin 1 ; genetics ; metabolism ; physiology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; Flow Cytometry ; Genetic Vectors ; chemistry ; genetics ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; pathology ; Lipids ; chemistry ; Microscopy, Confocal ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Phosphorylation ; Polymerase Chain Reaction ; methods ; Receptor, Epidermal Growth Factor ; metabolism ; Signal Transduction ; physiology ; Transfection ; methods

Objective To detect the clonal origin of intraductal proliferative lesions in the mammary gland. Methods Lesional and normal breast gland cells were microdissected from paraffin-embedded tissues using a laser capture microdissection (LCM) system. The genomic DNA was extracted. After digestion by restriction enzyme Hpa II, human androgen receptor (HUMARA) exonl was amplified by a fluorescent nested-PCR procedure and the PCR products were separated by a DNA sequencer and the fluorescent intensity of the two HUMARA alleles was analyzed. Results DNA from 88 of 101 (87.1%) patients could be amplified at the HUMARA locus and 68 of them (77.3%) were heterozygous and informative. 9/12 usual ductal hyperplasia (UDH) and 5/18 ductal intraepithelial neoplasia (DIN) 1A showed a polyclonal inactivation. 3/12 UDH, 13/18 DINIA, 28/28 DIN1B, 10/10 carcinoma in situ were monoclonal origin. Conclusion DIN 1A, 1B and carcinoma in situ are of monoclonal origin and real tumors.

Objective To detect the clonal origin of intraductal proliferative lesions in the mammary gland. Methods Lesional and normal breast gland cells were microdissected from paraffin-embedded tissues using a laser capture microdissection (LCM) system. The genomic DNA was extracted. After digestion by restriction enzyme Hpa II, human androgen receptor (HUMARA) exonl was amplified by a fluorescent nested-PCR procedure and the PCR products were separated by a DNA sequencer and the fluorescent intensity of the two HUMARA alleles was analyzed. Results DNA from 88 of 101 (87.1%) patients could be amplified at the HUMARA locus and 68 of them (77.3%) were heterozygous and informative. 9/12 usual ductal hyperplasia (UDH) and 5/18 ductal intraepithelial neoplasia (DIN) 1A showed a polyclonal inactivation. 3/12 UDH, 13/18 DINIA, 28/28 DIN1B, 10/10 carcinoma in situ were monoclonal origin. Conclusion DIN 1A, 1B and carcinoma in situ are of monoclonal origin and real tumors.