OBJECTIVEThe purpose of this study was to investigate the correlation between gene mutation and gene copy number and their association with the clinical profiles and pathological features in Chinese patients with non-small cell lung cancer (NSCLC).

METHODSSurgical specimens of cancer tissue were collected from 118 NSCLC patients. Gene mutations in exon 19 and exon 21 were detected by real-time PCR and gene copy number was detected by fluorescence in situ hybridization (FISH). Chi-square (χ(2)) test was performed to analyze the correlation between EGFR mutation and gene copy number, and explore their association with clinicopathological features in the NSCLC patients.

RESULTSThe mutation frequency in EGFR was 41.5% (49/118). EGFR mutations occured in 50.0% (48/96) of patients with adenocarinoma and 5.0% (1/20) of patients with squamous cell carcinoma. EGFR gene high copy number was detected in 70.3% (83/118)of the patients. The FISH-positive rate was 78.1% (75/96) in adenocarcinoma and 35.0% (7/20) in squamous cell carcinoma. EGFR mutation and high copy number mainly occurred in the adenocarcinoma, advanced stage, female gender, and non-smoking patients. There was a significant correlation between EGFR gene mutation and gene high copy number.

CONCLUSIONSEGFR gene mutation and gene high copy number are more common in Chinese NSCLC patients with adenocarcinomas, advanced stage, non-smokers and females. There is a significant correlation between gene mutation and gene high copy number. Combined analysis of EGFR mutation and gene copy number by FISH may provide a superior approach in selecting patients who may benefit from anti-EGFR target therapy.

Adenocarcinoma ; genetics ; pathology ; Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Carcinoma, Non-Small-Cell Lung ; genetics ; pathology ; Carcinoma, Squamous Cell ; genetics ; pathology ; Exons ; Female ; Gene Dosage ; Genes, erbB-1 ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Mutation ; Mutation Rate ; Neoplasm Staging ; Sex Factors ; Smoking

Objective To explore the dosage and injection method of concanavalin A(Con A) for inducing Wistar rats into the acute hepatic injury model. Methods (1)According to the dosage of Con A, 42 Wistar rats were randomly divided into groups A, B, C, D, E, N, 7 rats in each group. Group N was given tail intravenous injection of normal saline as normal control group. Groups A, B, C, D, E were given intravenous injection of 4, 8, 16, 30, 40 mg/kg of Con A respectively. At the 8th hour after modeling, the levels of alanine transaminase(ALT), aspartate aminotransferase(AST), albumin(ALB), interleukin(IL)-2 , IL-10, interferon (IFN)-γ, and tumor necrosis factor(TNF)-αwere detected. And HE staining was used to observe the pathological feature of hepatic tissue. (2)According to the injection method of Con A, 21 Wistar rats were randomly divided into normal control group, intraperitoneal injection group and tail intravenous injection group, 7 rats in each group. The dosage of Con A for the rats in intraperitoneal injection group and tail intravenous injection group was 16 mg/kg. At the 8th hour after modeling, the levels of serum ALT, AST, and ALB were determined. Results The number of abnormal deaths in various dose Con A groups at the end of each experiment was 0 in groups A, B, C, and 2 in group D, and 7 in group E. A small amount of spotty necrosis, inflammatory cell infiltration, and hepatic lobule with almost integrity of structure were found in groups A, B, while obvious bridging-like necrosis was seen in groups C, D. Serum ALT, AST, and ALB levels in intraperitoneal injection group had no statistically significant difference as compared with the normal control group. Conclusion Tail intravenous injection of 16 mg/kg of Con A can be used to induce an acute immunological liver injury rat model successfully.

OBJECTIVETo study the effect of the sphingosine kinase 1 (SphK1) inhibitor N,N-dimethylsphingosine (DMS) in combination with chemotherapeutic drugs (DDP, 5-Fu, MMC) on the proliferation of gastric cancer cells (SGC7901) in vitro, and to evaluate whether SphK1 inhibitors could be used as synergetic agents in chemotherapy.

METHODSSGC7901 cells were incubated in vitro with DMS (1 micromol/L) and 5-Fu, DDP, MMC at different concentrations in combination or separately for 24 h. The effects on the growth and survival of SGC7901 cells were determined by MTT assay. The inhibition rates were assessed by response surface analysis and the interactive relationships between the combined drugs were evaluated on the basis of positive/negative values of the cross product coefficients in the response surface equation.

RESULTSThe growth inhibition rate of the gastric cancer cells by treatment with DMS (1 micromol/L) was (10.23 +/- 0.74)%. The growth inhibition rates of the gastric cancer cells treated with 5-Fu (1, 5 and 25 microg/ml) for 24 h were (9.95 +/- 3.24)%, (21.04 +/- 2.19)%, and (45.49 +/- 3.60)%, respectively. The growth inhibition rates of the gastric cancer cells treated with DDP (0.5, 2.5 and 12.5 microg/ml) for 24 h were (9.38 +/- 0.79)%, (19.61 +/- 0.90)%, and (29.83 +/- 0.54)%, respectively. The growth inhibition rates of the gastric cancer cells treated with MMC (0.1, 0.5 and 2.5 microg/ml) for 24 h were (15.35 +/- 0.77)%, (24.72 +/- 0.83)%, and (30.68 +/- 0.28)%, respectively. There were significant differences among the inhibition rates caused by different concentrations of the drugs (P < 0.05). When 1 micromol/L DMS was used in combination with 5-Fu (1, 5, and 25 microg/ml) for 24 h, the growth inhibition rates of the cancer cells were (16.76 +/- 0.41)%, (27.28 +/- 0.29)% and (52.56 +/- 3.60)%, respectively. When 1 micromol/L DMS was used in combination with DDP (0.5, 2.5, and 12.5 microg/ml) for 24 h, the growth inhibition rates of the cancer cells were (15.35 +/- 0.86)%, (25.57 +/- 0.27)%, (36.37 +/- 0.51)%, respectively. When 1 micromol/L DMS was used in combination with MMC (0.1, 0.5, and 2.5 microg/ml) for 24 h, the growth inhibition rates of the cancer cells were (21.02 +/- 0.28)%, (32.10 +/- 0.27)%, (36.36 +/- 0.28)%, respectively. There were also significant differences among the growth inhibition rates caused by different concentrations of the drugs alone and in combination groups (P < 0.05).

CONCLUSIONSDMS can suppress the proliferation of SGC7901 cells in vitro, and there are evident synergetic effects when it is used in combination with chemotherapeutic drugs. The results of this study indicate that SphK1 inhibitors may become novel and promising chemotherapeutic sensitizers.

Antibiotics, Antineoplastic ; pharmacology ; Antimetabolites, Antineoplastic ; pharmacology ; Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Drug Synergism ; Enzyme Inhibitors ; pharmacology ; Fluorouracil ; pharmacology ; Humans ; Mitomycin ; pharmacology ; Phosphotransferases (Alcohol Group Acceptor) ; antagonists & inhibitors ; Sphingosine ; analogs & derivatives ; pharmacology ; Stomach Neoplasms ; pathology

Objective To explore the application of modified seldinger technique (MST) in advanced cancer patients and its superiority.Methods The MST was used in 20 patients who required peripherally insert central catheter (PICC) and they were divided into experimental group (n=40) and control group (n=40). The experimental group was treated with seldinger technique and the control group was treated with traditional PICC. The intubation success rate, complication rate and clinical application advantages were accounted and compare between the two groups.Results The intubation success rate in experimental group was 97.5% and that in control group was 80.0%,there were statistical significance (χ2=6.135,P<0.05); the incience of postoperative complications between experimetal group and control group were statistical significance (χ2=12.468,16.970,2.051;P<0.05).Conclusions Modified Seldinger technique with ultrasound guided have a useful effect, it should be widely carried out in clinical.

Capsid ; Cell Line ; Dependovirus ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Hepatitis B virus ; genetics ; Humans ; Liver Diseases ; therapy ; Plasmids ; Recombination, Genetic ; Transfection

Objective To investigate the effects of blue light on the thickness of corneal epithelium and full-thickness of the cornea in mice by optical coherence tomography angiography (OCTA).Methods Totally 40 mice were collected and randomly divided into experimental group and control group,with 20 mice in each group,and the experimental mice were raised in the blue light environment from 8 to 16 hours per day,while the controls were reared in normal environment.Then the thickness of corneal epithelium and full-thickness of the cornea in both groups were measured by OCTA before irradiation and one week,two weeks,one month,two months and three months after irradiation,respectively.Results Compared with pre-irradiation,the thickness of corneal epithelium of all regions did not change significantly in both groups at 1 week,2 weeks,and 1 month after irradiation,and the differences were not statistically significant (all P ＞ 0.05).Compared with before irradiation,the corneal epithelium thickness of the control group at 2 months and 3 months after irradiation did not change significantly,and there was no significant difference (both P ＞ 0.05).Compared with the control group,the corneal epithelium at central,nasal 5 mm,inferior 5 mm,and temporal 5 mm regions in the experimental group were significantly thickened,and the differences were statistically significant (all P ＜0.05).Three months after irradiation,compared with the control group,the thickness of corneal epithelium in the central and inner regions of the cornea and nasal 6 mm and temporal 6 mm regions of the experimental group were significantly thickened,and the differences were statistically significant (all P ＜ 0.05).There was no significant change in the corneal full thickness between the experimental group and the control group before irradiation and 1 week,2 weeks,1 month,2 months,and 3 months after irradiation,and the differences were not statistically significant (all P ＞ 0.05).Furthermore,the difference in the extremum value of corneal epithelial thickness,namely the maximum and the minimum,was significantly different in both groups (P ＜ 0.05),but the difference in the extremum value of the full-thickness of the cornea was not significant in the two groups (P ＞ 0.05).Conclusion The blue light can change the thickness of corneal epithelium in mice,and the change of the central region is obvious,but the full-thickness of the cornea do not significantly change in a short term.

OBJECTIVEInflammatory reaction and injury in immature lungs are associated with activation of nuclear factor-kappa B (NF-kappaB) to trigger proinflammatory cytokine release, but the mechanism thereof is not fully understood. The present study was conducted to understand possible relationship between expression of NF-kappaB and its inhibitor and severity and outcome of neonates with hyaline membrane disease (HMD).

METHODSSerial samples of bronchoalveolar lavage fluid (BALF) were obtained during mechanical ventilation from 31 preterm infants with HMD. These infants were divided into two groups: survivors group [n = 22, birth weight (1500 +/- 320) g and gestational age (31.2 +/- 1.8) weeks] and nonsurvivors group [birth weight (1340 +/- 280) g, gestational age (30.8 +/- 2.1) weeks]. Nineteen preterm infants [birth weight (1470 +/- 280) g, gestational age (30.6 +/- 1.9) weeks] without respiratory disorders were enrolled as control subjects. Alveolar macrophages (AM) were isolated by differential adherence. AM was cultured and treated with lipopolysaccharide (LPS) for 1 hr. Then, nuclear extracts of AM were analyzed by electrophoretic mobility shift assay (EMSA) for NF-kappaB expression. NF-kappaB inhibitor (IkappaB-alpha protein) in cytoplasmic extracts was detected by using Western blotting and IL-1beta and IL-8 in BALF by enzyme-linked immunosorbent assay (ELISA).

RESULTSNF-kappaB complexes were observed by EMSA, they were characterized by competition with cold oligonucleotide and p65-specific antibodies. The addition of an excess of cold oligonucleotide, corresponding to the NF-kappaB binding site, turned off the signal of the band, showing that the band was specific. An excess of an irrelevant oligonucleotide (corresponding to the SP-1) did not show any effect. The addition of an anti-p65 antibody caused the supershift of the two upper bands. After EMSA, the NF-kappaB complexes were quantified by using a ImageQuant software. NF-kappaB expression in AM at 24 hrs was higher in all the patients with HMD as compared with control subjects (survives/control, 34.1 vs 11.4 RDU, P < 0.01; nonsurvivors/control, 55.2 vs 11.4 RDU, P < 0.01). The NF-kappaB expression in AM at 72 hrs was higher than that in control subjects but not for nonsurvivors (survivors/control, 47.8 vs 25.6 RDU, P < 0.01; nonsurvivors/control, 21.8 vs 25.6, P > 0.05). The NF-kappaB expression in AM from nonsurvivors was depressed at 72 hrs as compared to 24 hrs (21.8 vs 55.2, P < 0.01), whereas the NF-kappaB expression in AM from survivors was still higher at 72 hrs than that at 24 hrs (47.8 vs 34.1, t = 4.43, P < 0.01).

CONCLUSIONAltered NF-kappaB activation in AM of BALF of neonates with HMD was observed, and it may be mediated by decreased IkappaB synthesis, increased IkappaB degradation, or both. In HMD nonsurvivors NF-kappaB translocation was hampered upon LPS activation.

Birth Weight ; Blotting, Western ; Bronchoalveolar Lavage Fluid ; cytology ; Cell Culture Techniques ; Cell Nucleus ; drug effects ; metabolism ; Cytoplasm ; drug effects ; metabolism ; Electrophoretic Mobility Shift Assay ; Enzyme-Linked Immunosorbent Assay ; Female ; Gestational Age ; Humans ; Hyaline Membrane Disease ; immunology ; therapy ; I-kappa B Proteins ; immunology ; Infant, Newborn ; Infant, Premature ; immunology ; Interleukin-1beta ; immunology ; Interleukin-8 ; immunology ; Lipopolysaccharides ; pharmacology ; Macrophages, Alveolar ; drug effects ; immunology ; Male ; NF-KappaB Inhibitor alpha ; NF-kappa B ; immunology ; Respiration, Artificial ; Severity of Illness Index ; Time Factors

WU polyomavirus (WUPyV), a new member of the genus Polyomavirus in the family Polyomaviridae, is recently found in patients with respiratory tract infections. In our study, the complete genome of the two WUPyV isolates (FZ18, FZTF) were sequenced and deposited in GenBank (accession nos. FJ890981, FJ890982). The two sequences of the WUPyV isolates in this study varied little from each other. Compared with other complete genome sequences of WUPyV in GenBank (strain B0, S1-S4, CLFF, accession nos. EF444549, EF444550, EF444551, EF444552, EF444553, EU296475 respectively), the sequence length in nucleotides is 5228bp, 1bp shorter than the known sequences. The deleted base pair was at nucleotide position 4536 in the non-coding region of large T antigen (LTAg). The genome of the WUPyV encoded for five proteins. They were three capsid proteins: VP2, VP1, VP3 and LTAg, small T antigen (STAg), respectively. To investigate whether these nucleotide sequences had any unique features, we compared the genome sequence of the 2 WUPyV isolates in Fuzhou, China to those documented in the GenBank database by using PHYLIP software version 3.65 and the neighbor-joining method. The 2 WUPyV strains in our study were clustered together. Strain FZTF was more closed to the reference strain B0 of Australian than strain FZ18.
Adult ; Child, Preschool ; China ; Evolution, Molecular ; Genome, Viral ; genetics ; Genomics ; Humans ; Male ; Molecular Sequence Data ; Phylogeny ; Polyomaviridae ; genetics ; isolation & purification ; Sequence Analysis, DNA ; methods

OBJECTIVETo evaluate the relationship between coronary artery stenosis and internal carotid artery, vertebral and basilar artery stenosis using computed tomographic angiography.

METHODSThe imaging and clinical data of 84 patients who underwent coronary, head and cervical computed tomographic angiography in our hospital between September 2008 and June 2010 were retrospectively analyzed. Segment stenosis scoring was performed to quantify the degree of stenosis of coronary arteries. The relationship between the segment stenosis scoring and the degree of stenosis for internal carotid artery, vertebral and basilar artery was analyzed.

RESULTThe coronary segment stenosis scores were significantly correlated with the degree of stenosis for internal carotid artery, vertebral and basilar artery (r=0.450 and 0.475,P<0.05) .

CONCLUSIONThe degree of the stenosis of coronary artery is associated with the stenosis of internal carotid artery, vertebral and basilar artery, which can be confirmed by computed tomographic angiography."

Aged ; Angiography ; methods ; Carotid Artery, Internal ; diagnostic imaging ; Carotid Stenosis ; diagnostic imaging ; Coronary Stenosis ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods ; Vertebrobasilar Insufficiency ; diagnostic imaging

Epidermal growth factor receptor (EGFR) gene mutation and copy number are useful predictive markers that guide the selection of non-small cell lung cancer (NSCLC) patients for EGFR-targeting therapy. This study aimed to investigate the correlation between EGFR gene mutation and copy number and clinicopathologic characteristics of Chinese patients with NSCLC. NSCLC specimens collected from 205 patients between November 2009 and January 2011 were selected to detect EGFR gene mutations with real-time polymerase chain reaction (RT-PCR) and to detect EGFR gene copy number with fluorescence in situ hybridization (FISH). EGFR mutations primarily occurred in females, non-smokers, and patients with adenocarinomas (all P < 0.001). Tissues from 128 (62%) patients were FISH-positive for EGFR, including 37 (18%) with gene amplification and 91 (44%) with high polysomy. EGFR gene mutation was correlated with FISH-positive status (R = 0.340, P < 0.001). Multivariate analysis showed that not smoking (OR = 5.910, 95% CI = 2.363-14.779, P < 0.001) and having adenocarcinoma (OR = 0.122, 95% CI = 0.026-0.581, P = 0.008) were favorable factors for EGFR gene mutation. These results show a high frequency of EGFR FISH positivity in NSCLC tissues from Chinese patients and a significant relevance between EGFR gene mutations and FISH-positive status. Among the FISH-positive samples, EGFR gene mutation occurred more frequently in samples with gene amplification compared to those with high polysomy, suggesting that EGFR mutation and gene amplification should be used as clinical decision parameters to predict response to EGFR-targeting therapy.
Adenocarcinoma ; genetics ; metabolism ; Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; Female ; Gene Amplification ; Gene Dosage ; Humans ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; genetics ; metabolism ; Male ; Middle Aged ; Mutation ; Real-Time Polymerase Chain Reaction ; Receptor, Epidermal Growth Factor ; genetics ; metabolism ; Smoking