Brain ; pathology ; Caudate Nucleus ; pathology ; Child, Preschool ; Humans ; Huntingtin Protein ; Huntington Disease ; diagnosis ; genetics ; pathology ; Magnetic Resonance Imaging ; Male ; Nerve Tissue Proteins ; genetics ; Pedigree ; Trinucleotide Repeats ; genetics

Background and purpose:Vascular endothelial growth factor(VEGF) is a potent angiogenic mediator and angiogenesis has important effects on tumor growth and metastasis.The present study was to investigate the relationship between genetic polymorphism of VEGF and heredity risk factor of lung cancer.Methods:VEGF genotypes were determined by PCR-RFLP method in 171 patients with lung cancer and 172 healthy controls.Software PHASE 1.0 was used to construct the haplotypes of every individual.Unconditional logistic regression model was used to analyze the statistical association of genotypes or haplotypes in the two groups adjusted by gender and age. Results:Individuals with at least one-2578A allele had a significantly decreased risk of lung cancer compared with those carrying-2578CC genotype.When the analyses were stratified by gender,the combined-2578 CA and AA genotype,were associated with a considerably reduced risk of lung cancer(P=0.001,OR=0.303,95%CI=0.15 3-0.601).The distribution of the two haplotypes(936C/-2578C and 936C/-2578A) among overall lung cancer cases was significantly different from that among the controls(P=0.016,0R=0.317,95%CI=0.124-0.809 and P=0.018,OR=0.547, 95%CI=0.331-0.903).When the cases were categorized by tumor histology,the distribution of C-C haplotype in the adenocarcinoma(AC) group was associated with a substantially lowered risk of AC(P=0.004,0R=0.237,95%CI=0.090- 0.627),compared with the reference haplotypes.Conclusion:VEGF polymorphism may be a critical risk for the genetic risk factor to lung cancer.

OBJECTIVETo analyze the comorbidities in patients with cerebral palsy (CP) from two perspectives as neurologic subtype and gross motor functions, and find their correlations.

METHODSChildren with cerebral palsy treated in the rehabilitation center from January 2007 to June 2009 received the following examinations: intelligence capacity test, ophthalmologic consultation, language-speech test, brainstem auditory evoked potential and electroencephalogram. They were stratified according to both neurologic subtype and gross motor functions to detect the occurrence of comorbidities.

RESULTSOf all the 354 cases, 166 (46.89%) had mental retardation, 15 (4.24%) auditory limitations, 138 (38.98%) visual disorder, 216 (61.02%) language-speech disorder and 82 (23.16%) epilepsy. The frequency of individual comorbidities were distributed disproportionately between the different neurologic subtypes. Correlation analysis showed that there was a significant correlation between the spastic diplegia and the visual disorder (correlation coefficient = 0.26), between spastic hemiplegia and epilepsy (correlation coefficient = 0.17), between spastic quadriplegia and epilepsy and mental retardation (the correlation coefficient was 0.38 and 0.11, respectively) and between both dyskinetic and mixed children and language-speech disorder (the correlation coefficient was 0.24 and 0.27, respectively). The frequency of individual comorbidities was distributed disproportionately between the different neurologic subtypes and between the different GMFCS levels (P < 0.05), except for the frequency of visual disorders (chi(2) = 1.90, P > 0.05); and with the increase of the GMFCS levels, the burden of the comorbidities were more heavy and the incidence of the comorbidities was higher. Multi-comorbidities were relatively infrequently encountered in those with spastic hemiplegic or spastic diplegic children or patients whose GMFCS levels were I-III, while these entities occurred at a frequent level for those with spastic quadriplegic, dyskinetic, or mixed or children whose GMFCS levels were IV and V, and the differences were significant (P < 0.05). The mean GMFCS levels of children with spastic quadriplegic, dyskinetic or mixed CP were higher than level III, most of them had no ability of ambulation;while the mean GMFCS levels of spastic hemiplegic or spastic diplegic children were below level III, most of them could walk independently.

CONCLUSIONSThere are correlations between the occurrence of the comorbidities such as mental retardation, auditory or visual impairments, language-speech disorders, epilepsy and the cerebral palsy subtype and the gross motor function levels. Clinicians should have a full recognition of these comorbidities, and we should have a cooperation between the different subjects to have an overall evaluation and rehabilitation and to improve the prognosis.

Adolescent ; Cerebral Palsy ; classification ; epidemiology ; Child ; Child, Preschool ; Comorbidity ; Epilepsy ; classification ; epidemiology ; Female ; Humans ; Infant ; Male ; Motor Skills ; classification ; Motor Skills Disorders ; classification ; epidemiology ; Quadriplegia ; classification ; epidemiology ; Vision Disorders ; classification ; epidemiology

OBJECTIVETo evaluate the correlation between standardized uptake valus (SUV) of 18F-fluorodeoxyglucose (18 F-FDG) of tumor at PET/CT examination and the expression of glucose transporter-1 (Glutl) and Ki-67 in esophageal cancer.

METHODS56 patients with esophageal cancer were evaluated with 18 F-FDG PET/CT examination before operation. The expression of Glut1 and Ki-67 antigen in the tumor tissues was detected by immunohistochemistry after operation.

RESULTS(1) Positive rate of Glutl and Ki-67 expression in esophageal cancer tissues was 100% , respectively. There was a positive correlation between the expression of Glutl and Ki-67 and the clinical stages and differentiation of the tumor. The more the tumor and the clinical stages were advanced and the lower was the tumor differentiation, the more Glutl and Ki-67 were expressed. (2) There were abnormal radioactive high uptake regions on PET/CT imaging of esophagus in the 56 patients, which were confirmed by pathology as the primary carcinoma. The SUV was higher than 2. 5. There was a gradually increasing tendency in SUV along with the lowering of the tumor differentiation and the advance of clinical stages. (3)There was a correlation between the expression of Glutl, Ki-67 and the SUV, the more Glutl and Ki-67 were expressed, the higher the SUV of tumor 18F-FDG at PET/CT examination was in esophageal tumor tissues.

CONCLUSIONThere is a widespread expression of Glutl in esophageal cancer tissues, and the SUV may be used to indirectly evaluate the proliferative capacity of esophageal cancer.

Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Differentiation ; Esophageal Neoplasms ; metabolism ; pathology ; Female ; Fluorodeoxyglucose F18 ; pharmacokinetics ; Glucose Transporter Type 1 ; metabolism ; Humans ; Immunohistochemistry ; Ki-67 Antigen ; metabolism ; Male ; Middle Aged ; Neoplasm Staging ; Positron-Emission Tomography ; methods ; Tissue Distribution ; Tomography, X-Ray Computed ; methods

OBJECTIVETo evaluate the possibility of hepatitis B virus (HBV) as a vector in liver-targeting gene therapy.

METHODSA fragment containing the small envelope gene of HBV was replaced with the reporter gene green fluorescent protein (GFP) to construct the recombinant HBV vector, which was transfected into HepG2 cells with liposome. The expression of GFP was observed with fluorescence microscope. The HBV cccDNA was testified using semi-nest PCR. The viral particles of the recombinant HBV in culture medium were detected by PCR as well as Southern blot.

RESULTSThe HBV vector carrying the interesting gene of GFP could express the functional protein in the transfected hepatocytes. However, the recombinant HBV vector was replication-deficient, which could not be packed and replicated in the hepatocytes to secrete mature recombinant HBV particles carrying the interesting gene of GFP when transfected solely but could when cotransfected with the recombinant and helper construct which lacked part of 5'-proximal HBV RNA packaging signal epsilon.

CONCLUSIONIt is possible that HBV is reconstructed as a liver-targeting vector for gene therapy.

Cell Transformation, Viral ; Cells, Cultured ; Gene Transfer Techniques ; Genes, Reporter ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; physiology ; Hepatitis B virus ; genetics ; physiology ; Hepatocytes ; cytology ; virology ; Humans ; Liver ; cytology ; virology ; Recombinant Proteins ; genetics ; Transfection ; Virus Replication

Animals ; Carbon Tetrachloride ; adverse effects ; Chemical and Drug Induced Liver Injury, Chronic ; prevention & control ; Chronic Disease ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Mice ; Mice, Inbred BALB C

OBJECTIVETo explore the possibility of using HBV as a gene delivery vector, and to test the anti-HBV effects by intracellular combined expression of antisense RNA and dominant negative mutants of core protein.

METHODSFull length of mutant HBV genome, which expresses core-partial P fusion protein and/or antisense RNA, was transfected into HepG2.2.15 cell lines. Positive clones were selected and mixed in respective groups with hygromycin in the culture medium. HBsAg and HBeAg, which exist in the culture medium, were tested by ELISA method. Intracellular HBc related HBV DNA was examined by dot blot hybridization. The existence of recombinant HBV virion in the culture medium was examined by PCR. Free of packaging signal, HBV genome, which express the HBV structural proteins including core, pol and preS/S proteins, was inserted into pCI-neo vector. HepG2 cell lines were employed to transfect with the construct. G418 selection was done at the concentration of 400mug/ml in the culture medium. The G418-resistant clones with the best expression of HBsAg and HBcAg were theoretically considered as packaging cell lines and propagated under the same conditions. It was transfected with plasmid pMEP-CPAS and then selected with G418 and hygromycin in the culture medium. The existence of recombinant HBV virion in the culture medium was examined by PCR.

RESULTSThe mean inhibitory rates of HBsAg were 2.74% 3.83%, 40.08 2.05% (t=35.5, P<0.01), 66.54% 4.45% (t=42.3, P<0.01), and 73.68% 5.07% (t=51.9, P<0.01) in group 2.2.15-pMEP4, 2.2.15-CP, 2.2.15-SAS, and 2.2.15-CPAS, respectively. The mean inhibitory rates of HBeAg were 4.46% 4.25%, 52.86% 1.32% (t=36.2, P<0.01), 26.36% 1.69% (t=22.3, P<0.01), and 59.28% 2.10% (t=39.0, P<0.01), respectively. The inhibitory rates of HBc related HBV DNA were 0, 82.0%, 59.9%, and 96.6%, respectively. Recombinant HB virion was detectable in the culture medium of all the three treatment groups. G418-resistant HBV packaging cell line, which harbored an HBV mutant whose packaging signal had been deleted, was generated. Expression of HBsAg and HBcAg was detectable. Transfected with plasmid pMEP-CPAS, it was found to secrete recombinant HB virion and no wild-type HBV was detectable in the culture medium.

CONCLUSIONSIt has stronger anti-HBV effects by combined expression of antisense RNA and dominant negative mutants than by individual expression of them. With the help of wild-type HBV, the modified HBV genome can form and secret HBV like particles, which provides evidence that the antiviral gene will be hepatotropic expression and the antiviral effects will be amplified. The packaging cell line can provide packaging for replication-defective HBV, but with low efficiency.

Cell Line ; Genetic Engineering ; Genetic Vectors ; Hepatitis B virus ; genetics ; Mutation ; Plasmids ; RNA, Antisense ; physiology ; Transfection ; Viral Core Proteins ; physiology ; Virus Assembly ; Virus Replication

OBJECTIVETo cooperate with the study of HBV vector, hygromycin-resistant packaging cell line was developed that allows encapsidation of plasmids into HBV particles.

METHODSFree of packaging signal, HBV genome was inserted into plasmid pMEP4, which expresses the HBV structural proteins including core, pol and preS/S proteins. HepG2 cell lines were employed to transfect with the construct. Hygromycin selection was done at a concentration of 150 micrograms/ml in the culture medium. The hygromycin-resistant clones with the best expressions of HBsAg and HBcAg were theoretically considered as packaging cell line and propagated under the same conditions. It was infected with recombinant retrovirus vector and hen selected with G418 and hygromycin in the culture medium. The existence of recombinant HBV virion in the culture medium was examined by PCR.

RESULTSHygromycin-resistant HBV packaging cell line was generated, which harbored an HBV mutant whose packaging signal had been deleted. Expressions of HBsAg and HBcAg were detectable. Infected with recombinant retrovirus pRV-CP, the hygromycin-resistant packaging cell line was found to secrete mutant HBV particles and no wild-type HBV was detectable in the culture medium.

CONCLUSIONAfter the packaging signal was deleted and transfected into HepG2 cell lines, the partial HBV genome lost its ability to form wild-type HBV, but conserves cis-action providing structural proteins for the packaging of the replication-defective HBV.

Cell Line ; Drug Resistance, Viral ; Genetic Vectors ; Genome, Viral ; Hepatitis B virus ; drug effects ; genetics ; Humans ; Hygromycin B ; pharmacology ; Mutation ; Plasmids ; Retroviridae ; genetics ; Transfection ; Virus Assembly

BACKGROUNDTo explore the possibility of using HBV as a gene delivery vector, and to test the anti-HBV effects by intracellular expression of dominant negative mutants of core protein.

METHODSTwo kinds of full length mutant HBV genome, which express Core-partial P and Core-S fusion protein, were transfected into HepG 2.2.15 cell lines. Positive clones were selected and mixed in respective groups with hygromycin in the culture medium. HBsAg and HBeAg, which exist in the culture medium, were tested by ELISA and intracellular HBc related HBV DNA was examined by dot blot hybridization. The existence of recombinant HBV virion in the culture medium was examined by PCR.

RESULTSThe mean inhibitory rates of HBsAg were 2.74+/-3.83%, 40.08+/-2.05% (P less than 0.01) and 52.94+/-1.93% (P less than 0.01) in group 2.2.15-pMEP4, 2.2.15-CP and 2.2.15-CS, respectively. The mean inhibitory rates of HBeAg were 4.46+/-4.25%, 52.86+/-1.32% (P less than 0.01) and 41.60+/-1.65% (P less than 0.01), respectively. The inhibitory rates of HBc related HBV DNA were 15.3%, 82.0% and 67.2%, respectively. Recombinant HBV virion was detectable in the culture medium of only group 2.2.15-CP.

CONCLUSIONDominant negative mutants of core protein can efficiently suppress wt-HBV replication and the expressions of HBV antigens. With the help of wild-type HBV, the recombinant HBV genome can form and secret HBV like particles, which provides evidence that the antiviral gene will be hepatotropic expression and the antiviral effects will be amplified.

Cell Line, Tumor ; Genetic Engineering ; Genetic Therapy ; Genetic Vectors ; Genome, Viral ; Hepatitis B Core Antigens ; biosynthesis ; physiology ; Hepatitis B virus ; genetics ; Humans ; Point Mutation ; Recombinant Fusion Proteins ; biosynthesis ; physiology ; Virus Replication

OBJECTIVESTo analyze the survival outcomes of the surgery for colorectal cancer with liver metastases (CRCLM), and study the mode of multi-disciplinary team (MDT) for CRCLM.

METHODSThe retrospective analysis was conducted for 38 patients with CRCLM received MDT management and surgical treatment from January 2009 to August 2011. The peri-operative and survival outcomes of MDT and surgery were evaluated.

RESULTSAll the cases met the present criteria of resetability for CRCLM, but only 4 cases (10.5%) met the previous one. Coloproctectomy and hepatectomy were performed in all cases, with 39 colorectal neoplasms and 155 liver lesions removed. One case died of postoperative septic shock. Colorectal and hepatic specific complications were absent in the others patients except one case of biliary leak which was treated with conservative management. Neoadjuvant chemotherapy was arranged in 13 cases. Adjuvant chemotherapy was administered for every patient. After a mean follow-up of (22 ± 10) months according to the finding time of liver metastases, recurrence and metastases were observed in 16 cases and 6 cases died of late-stage cachexia. The 1-, 2- and 3-overall survival rate were 94.4%, 85.3% and 75.8% respectively, and the 1-, 2- and 3-disease-free survival rate were 70.1%, 54.2% and 54.2% respectively.

CONCLUSIONSMDT mode for resectable CRCLM is recommendable. Surgical resection of CRCLM is feasible and safe, which seems to achieve favourable short-middle oncologic outcomes. And long-term survival is expected.

Adult ; Aged ; Chemotherapy, Adjuvant ; Colorectal Neoplasms ; mortality ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Liver Neoplasms ; mortality ; secondary ; Male ; Middle Aged ; Retrospective Studies ; Survival Rate ; Treatment Outcome