Carcinoma ; genetics ; metabolism ; pathology ; surgery ; China ; Humans ; Male ; Oncogene Proteins, Fusion ; genetics ; Prostatic Hyperplasia ; genetics ; metabolism ; pathology ; surgery ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Proto-Oncogene Proteins c-ets ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Serine Endopeptidases ; genetics ; metabolism

OBJECTIVETo investigate DNA aneuploid and P16 expression in biopsy specimens from lung cancer, and to study genetic instability and the application of flow cytometry in lung cancer pernicious degree diagnosis.

METHODSBlood cells and cancer cells in biopsy specimens were marked simultaneously with anti-CD45 and anti-P16 fluorescent antibody, and the ratio of CD45+ P16+ cells and CD4- P16+ cells was compared. DNA content in biopsy specimens from lung cancer was detected by flow cytometry.

RESULTSAmong the 74 cases of lung cancer, there are 46 cases of DNA aneuploid (62.2%). Thirty-seven cases of lung cancer expressed P16 lowly (50%). Twelve cases of lung cancer only expressed P16 lowly (16.22%), 21 cases of lung cancer only expressed DNA aneuploid (28.38%), and 25 cases not only expressed P16 lowly but also expressed DNA aneuploid (33.78%). Indexes of malign degree, such as P16 low expression or DNA aneuploid could be detected in 58 cases among the 74 cases (78.38%) by flow cytometry.

CONCLUSIONP16 low expression and DNA aneuploid are the indexes of lung cancer malign degree, and flow cytometry can be used to study genetic instability and evaluate biopsy specimens from lung cancer.

Aneuploidy ; Animals ; Biopsy ; Chromosomal Instability ; genetics ; DNA ; genetics ; Female ; Flow Cytometry ; Gene Dosage ; Gene Expression Regulation, Neoplastic ; Genes, p16 ; Humans ; Leukocyte Common Antigens ; genetics ; Lung Neoplasms ; diagnosis ; genetics ; pathology ; Male ; Mice ; Middle Aged

OBJECTIVETo explore the influence of inhibition of hypoxia-inducible factor-1 alpha (HIF-1 alpha) by RNA interference (RNAi) on tumorigenesis of human myeloma cell line (HMCL) RPMI8226 cells in nude mice.

METHODRNAi vector of HIF-1 alpha was constructed with commercial shRNA expression vector pSilencer 2. 1-U6 hygro. RT-PCR and western blot were used to detect HIF-1 alpha mRNA and protein expression respectively. Vascular endothelial growth factor (VEGF) secretion and cell cycle changes were detected by ELISA and flow cytometry respectively. Expression of target gene of HIF-1 alpha, VEGF and Glut-1 were tested under hypoxia condition. Tumorigenesis was observed after transfected cells were injected subcutaneously in nude mice.

RESULTSAfter interference, expression of HIF-1 alpha decreased significantly at both mRNA and protein level. Under normoxia condition, VEGF concentrations in HIF-la inhibited cells (RPMI8226-il and RPMI8226-i2) and non-inhibited cells (RPMI8226-c and RPMI8226) showed no differences. While under hypoxia condition, VEGF concentration in the above four cells was (506.0 +/- 53.2), (494.7 +/- 63.1), (984.4 +/- 61.9) and (938.2 +/- 62.2) pg/ml, respectively, being significantly lower in RPMI8226-il and RPMI8226-i2 cells than in RPMI8226-c and RPMI8226 cells (P <0.05). HIF-1 alpha interference was found to suppress the cells shift from S-phase to G1 induced by hypoxia. VEGF and Glut-1 expressions were markedly attenuated (P <0.05). The growth rate of HIF-1 alpha inhibition tumors in subcutaneous xenograft model decreased drastically.

CONCLUSIONSRNAi inhibits HIF-1 alpha expression. Reduced tumor growth by HIF-1 alpha inhibition may partly through inhibiton of angiogenesis and glycolysis metabolism.

Animals ; Cell Cycle ; Cell Line, Tumor ; Genetic Vectors ; Glucose Transporter Type 1 ; metabolism ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Mice ; Mice, Nude ; Multiple Myeloma ; metabolism ; pathology ; RNA Interference ; Vascular Endothelial Growth Factor A ; metabolism ; Xenograft Model Antitumor Assays

OBJECTIVETo detect the serum proteomic fingerprints in patients with hypopharyngeal squamous cell carcinoma (HPSCC) by surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) protein chip array technique.

METHODSThe serum samples were obtained from 58 HPSCC patients for protein expression analysis using SELDI-TOF Protein Chip technique and cation-exchange (CM10) protein array. All the spectra were compared and the qualified mass peaks with mass-to-charge ratios (m/z) between 1 and 70 kD were autotimatically detected. The tree analysis pattern was generated using Biomarker Patterns Software.

RESULTSThe protein profiles of HPSCC serum were analyzed according to the clinical and pathological features of the patients and their treatment response. No significant difference was noted in the serum proteins between HPSCC patients with different statuses of cervical lympha node metastasis (P>0.05), and the difference between well differentiated and poorly differentiated HPSCC was only minor. No significant difference was found in the serum proteins between chemotherapy-sensitive patients and the insensitive patients (P>0.05), but 5 proteins were identified to be overexpressed in the sensitive patients (P < / = 0.05). Radiotherapy-sensitive HPSCC patients were segregated from the insensitive group with a sensitivity of 86.67% and specificity of 100%.

CONCLUSIONThe serum protein at the m/z value of 6115.74 is overexpressed in radiotherapy-sensitive HPSCC patients. Serum protein profiling allows the prediction of radiotherapy response in HPSCC patients, and the identified proteins may serve as candidate biomarkers for predicting the radiotherapy sensitivity of HPSCC.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; analysis ; Carcinoma, Squamous Cell ; genetics ; radiotherapy ; Female ; Humans ; Hypopharyngeal Neoplasms ; genetics ; radiotherapy ; Male ; Middle Aged ; Models, Biological ; Proteome ; analysis ; Radiation Tolerance ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

OBJECTIVETo investigate the expression of calcium/calmodulin-dependent serine protein kinase (CASK) induced by short-term hypoxia, and to explore the role of JNK pathway in this signal event.

METHODSEA. hy926 cells were cultured in normoxic condition for 0, 12, 24, 48, 72 h after being exposed to hypoxic condition for 3 h, then the cellular lysates were extracted. CASK promoter luciferase reporter recombinant was constructed and transfected into EA. hy926 cells for 48h. Cellular lysates were extracted 1, 3, 6, 12 h after hypoxia treatment and were used to detect firefly luciferase activity and rinella luciferase activity with luminometer. EA. hy926 cells were cultured under hypoxic condition for 1, 3, 6, 12 h or under normoxic condition, then the cell lysates were extracted and used to detect phospho-JNK with Western blot. EA. hy926 cells were pretreated with different concentrations of JNK specific inhibitor SP 600125 (0, 10, 100 nmol/L and 1,10 micromol/L) 1h before hypoxic treatment of various duration, and the cell lysates were extracted to detect CASK expression with Western blot.

RESULTSCASK expression was obviously elevated by hypoxia, and the high expression sustained for 72 h when the hypoxic cells were cultured in normal conditions, and it was significantly higher than that of normal controls. Dual luciferase reporter assay showed that CASK promoter activity was significantly increased after hypoxia (0.010 +/- 0.003, P < 0.01), and it reached the peak 12 hrs after hypoxia (0.192 +/- 0.023, P < 0.01). The phosphorylation of JNK was enhanced with the prolongation of hypoxic time. CASK protein expression was suppressed by JNK specific inhibitor SP600125 in a dose dependent manner, and it decreased to the lowest level with 10 micromol/L SP600125 pretreatment.

CONCLUSIONJNK signal pathway is involved in short-term hypoxia related CASK upregulation.

Calcium ; metabolism ; Cell Hypoxia ; Cell Line ; Endothelial Cells ; metabolism ; physiology ; Guanylate Kinases ; metabolism ; Humans ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Signal Transduction

OBJECTIVETo investigate the influence of hypoxia on the proliferation and activity of human umbilical vein vascular endothelial cells (EA. hy926).

METHODSEA. hy926 cells were cultured in vitro and divided into normal control and hypoxia groups. The cells in hypoxia group were placed into hypoxic jar and treated with mixed gases(94% N2 +5% CO2 + 1% O2) for 1,3,6 and 12 hours. Then the total proteins were extracted for the determination of the expression of vascular endothelial growth factor (VEGF) and proliferation cell nuclear antigen (PCNA). The cell cycle and growth curve were determined with flow cytometry and MTT method, respectively.

RESULTSThe expression of PCNA protein began to increase at 3 post-hypoxia hour (PHH), peaked at 6 PHH, but without obvious difference compared with that at 12 PHH. The expression of VEGF began to increase at 1 PHH, peaked at 6 PHH, and decreased at 12 PHH, though it was still markedly higher than that of normoxia at 12 PHH. MTT results showed that the cell activity began to increase at 1 PHH, and it was still to increased at 3 PHH, then decreased at 6 PHH, and it was lower than that in control group at 12 PHH. The number of cells in G0/G1 phase was decreased, but the cells in S and G2/M phase was increased at 1, 3, 6 PHH when compared with those in normal controls. The proliferation index (PI) of cells in hypoxia group at 1PHH (43 +/- 9)%, 3PHH (39 +/- 11)%, 6 PHH (40 +/- 11))% were higher than that before hypoxia (32 +/- 9)% and 3 (39 +/- 11) % and 6 hours (40 +/- 11)% after hypoxia (P < 0.05). The PI was obviously lower at 12 PHH (27 +/- 4))% compared with that of cells under normoxic condition (P < 0.05).

CONCLUSIONShort-term hypoxia is beneficial to promote the proliferation of the cells, but this effect will be inhibited with the prolongation of hypoxia.

Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Endothelial Cells ; metabolism ; Humans ; Hypoxia ; metabolism ; pathology ; Proliferating Cell Nuclear Antigen ; metabolism ; Umbilical Veins ; cytology ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the influence of human telomerase reverse transcriptase (hTERT) gene transfection on the proliferation of human embryonic fibroblasts (hEF).

METHODShEFs were cultured in vitro. Sense recombinant eukaryotic plasmid (pIRES2-EGFP-hTERT) and pIRES2-EGFP vacant vector were transfected into hEF respectively with Lipofectin reagent, and were named as hEF-hTERT and hEF-EGFP. The hTERT, Id1, PCNA and I, III type collagen expression in these cells were detected by Western blot. Then the cell cycle and growth curve were measured and plotted with flow cytometry and MTT method, respectively.

RESULTS1. The expression of hTERT, Id1, PCNA, type I and III collagen in hEF-hTERT were much higher than that in hEF and hEF-EGFP. 2. As shown in the growth curve, the OD value of hEF-hTERT at 4 to 6 days after culture was obviously higher than that of hEF and hEF-EGFP (P < 0.05), while no difference existed between hEF and hEF-EGFP from 1 to 6 days after culture (P > 0.05). 3. The cell number in G0/G1 phase in hEF-hTERT was less than that in hEF and hEF-EGFP. The cell number of hEF-hTERT in S and G2/M phase and its proliferation index (57.47%) increased when compared with that in hEF-EGFP (13.13%) and hEF (17.38%), but there was no difference between hEF and hEF-EGFP.

CONCLUSIONExogenous hTERT gene transfection could promote the proliferative capacity of hEF.

Cell Proliferation ; Cells, Cultured ; Embryonic Stem Cells ; cytology ; Fibroblasts ; cytology ; Humans ; Telomerase ; genetics ; Transfection

OBJECTIVETo probe into the effects of familial factors on injury-related behaviors in children.

METHODSInjury-related behaviors and familial factors of 6884 children were investigated with Family Questionnaire and Child Behavior Checklist. Multi-nominal logistic regression analysis was performed.

RESULTSThere were 1670 (24.26%) children having serious injury-related behaviors and 3683 (53.50%) children having moderate injury-related behaviors. Factors contributing to children's injury-related behaviors would include punishment or indifference as well as the mode of parents' education; reintegral type of family; the level of parents' cognition on injuries; unfit location of medicine at home and careless attitudes of parents.

CONCLUSIONThere was close relationship between children's injury-related behaviors and familial factors. To avoid injury-related behaviors and to prevent injury occurrence, the importance of familial factors must be stressed.

Child ; Child Behavior ; psychology ; Family ; psychology ; Female ; Humans ; Logistic Models ; Male ; Surveys and Questionnaires ; Wounds and Injuries ; etiology ; prevention & control ; psychology

OBJECTIVESTo investigate the effect of four types of neck dissections for the recurrent and persistent lymph nodes of NPC after radiotherapy.

METHODSThe clinical data of 88 cases of nasopharyngeal carcinoma with recurrent and persistent lymph nodes after radiotherapy were analyzed retrospectively, the 5-year survival rate, recurrent rate, distant metastatic rate and surgical complications were analyzed. The survival rate and recurrent rate of the radical neck dissection (RND), modified radical neck dissection (MRND), selective neck dissection (SND) and lymph node resection (LNR) were compared. The survival and recurrent rate between those with and without postoperative radiotherapy were investigated as well.

RESULTSThe 5-year survival rate and recurrent rate of whole group were 42.8% and 22.7%, respectively. As for the patients with disease staged II, III, IV, the 5-year survival rates were 56.7%, 36.1% and 32.4%, respectively. The 5-year survival rate of groups of RND, MRND, SND and LMR were 39.8%, 60.0%, 37.9% and 44.1%, respectively, the differences were insignificant (Log Rank = 1.0, P = 0.8011), the recurrent rate of the lymph node among the 4 groups were insignificant either (chi2 = 0.470, P = 0.493). The 5-year survival rates of those with and without postoperative radiotherapy were 39.1% and 45.3%, respectively, the differences were insignificant (Log Rank = 0.06, P = 0.8138), the recurrent rate of the two groups were insignificant (chi2 = 0.593, P = 0.441).

CONCLUSIONSThe four types of neck dissection were effective and safe to control the recurrent and persistent lymph nodes in the neck after radiotherapy, as long as choosing patients rationally and gave postoperative radiotherapy if necessary.

Adult ; Female ; Humans ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; pathology ; surgery ; Neck Dissection ; methods ; Neoplasm Recurrence, Local ; surgery ; Neoplasm, Residual ; surgery ; Treatment Outcome

According to international medical information standard and Chinese healthcare management criterion,this paper study EMR features which focus on standard documents exchange and sharing.Tele Regional Healthcare Platform is established by EMR in order to realize medical resource sharing between big hospital in the situation of China, one solution is offered to stave difficulty and high expense of medical service in countryside.
Computer Communication Networks ; Medical Records Systems, Computerized ; Software Design ; Telemedicine ; instrumentation ; methods