Animals ; Apoptosis ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Cells, Cultured ; HSP70 Heat-Shock Proteins ; metabolism ; Heat Stress Disorders ; metabolism ; Myocytes, Cardiac ; metabolism ; Rats ; Rats, Wistar

ObjectiveTo screen the variations of the human protein kinase Cγ gene (PRKCG) and study their association with Parkinson's disease(PD).MethodsDNA was extracted from blood of patients with PD and matched normal controls. All 18 exons including the exon-intron junctions were amplified in 17 different PCR fragments, which were analyzed for the presence of variations by DHPLC. The PCR products with a heteroduplex peak were sequenced. Significance was evaluated from 2×2 contingency tables byX2 test on the basis of the total number of alleles at each locus. Case-control association analysis was performed between candidate polymorphisms and PD. ResultsIn the 50 early-onset PD(EOPD) patients and 50 controls, there was no missense mutation, insertions or deletions in coding regions of the PRKCG. But 2 different single nucleotide polymorphism(SNPs) in exons, 5 different SNPs and 1 tetranucleotide repeat in introns were identified. Five of them [IVS3+96G>T, IVS11+26T>G, IVS15-41T>C, IVS16-59G>A, IVS16-42(TCTG)1-2] were described here for the first time. Three of them (IVS11+26T>G, IVS13+76T>C,1497T>C),in complete linkage,constituted a haplotype block. In the preliminary association analysis, the frequency of IVS13+76C, IVS11+26G and 1497C allele on this haplotype block was significantly higher in EOPD patients than the controls (24% vs 9%)(X2=8.165,P=0.004,OR=3.193, 95%CI:1.400-7.282). But in a larger sample of 156 EOPD patients, 153 late-onset PD(LOPD) patients and 195 normal controls, there was no significant difference between the three groups (12.8%,13.7% ,14.6%)(X2=0.471,P=0.790). ConclusionThe PRKCG gene might not be a risk factor for sporadic PD.

Objective To investigate the relationship between tumor invasion and changes of mRNA expression of MMP-2, TIMP-2 and CD147 in human pituitary adenoma. Methods 60 patients with pituitary adenoma were devided into two groups, invasive and non-invasive group, by MRI. The expression level of MMP-2,TIMP-2 and CD147 in the samples of pituitary adenoma was measured by semiquantitative reverse transcriptasepolymerase chain reaction (RT-PCR). Pearson analysis was used to reveal the correlation between the expression of each marker. Results The mRNA expression level of MMP-2 and CD147 was significantly higher in invasive pituitary adenoma group than that in non-invasive group (P ＜ 0. 01 ) while the mRNA expression level of TIMP-2was lower in invasive pituitary adenoma group than that in non-invasive group (P ＜0.01 ). According to Pearson analysis, the mRNA expression of MMP-2 was positively correlated with CD147 in invasive pituitary adenoma (r=0. 69, P ＜ 0. 05 ), and M MP-2 was negatively correlated with TIMP-2 in non-invasive pituitary adenoma (r =-0.68, P ＜ 0.05 ). Conclusions The invasion of human pituitary adenoma are closely related to the low expression level of TIMP-2 as well as the high expression level of MMP-2 and CD147. MMP-2, TIMP-2 and CD147 can be used as indicators of tumor invasion of human pituitary adenoma.

Objective To investigate the surgical treatment strategies and prognostic factors of cT4bM0 primary rectal cancer.Methods The clinical data of 53 patients with cT4bM0 primary rectal cancer who were admitted to the First Hospital of Peking University from January 2000 to December 2010 were retrospectively analyzed.All the patients received en-bloc multivisceral resection and postoperative chemotherapy.The survival and prognostic factors were analyzed.The patients were followed up via out-patient examination,phone call or mail,and the follow-up was ended till December 2012.The survival curve was drawn using the Kaplan-Meier method,and the survival was analyzed using the Log-rank test.Uni-and multivariate analysis were done using chisquare test and COX's proportional hazard model.Results Of all the 53 patients,20 received posterior pelvic exenteration (PPE),20 received total pelvic exenteration (TPE),3 received low anterior resection (LAR) +local resection of ballder,2 received LAR + ovariectomy,2 received LAR + local resection of ureter,1 received LAR + local resection of posterior vaginal wall,1 received LAR + vesiculectomy and vesectomy,3 received abdominoperineal resection (APR) + local resection of posterior vaginal wall,1 received APR + sacrectomy.R0 resection was achieved in all the patients.No intraoperative death was observed,and the incidence of postoperative complication was 9.4％ (5/53).The results of postoperative pathological examination showed that 2 patients were with well-differentiated adenocarcinoma,41 with moderate-differentiated adenocarcinoma,and 10 with poorly differentiated adenocarcinoma.Twenty-four patients were with lymph node metastasis.Four patients were in TNM stage Ⅰ,25 in TNM stage Ⅱ and 24 in TNM stage Ⅲ.Fifty-three patients were followed up postoperatively,and the median time for follow-up was 33 months (range,4-116 months).The overall 5-year survival rates was 57.3％.The 5-year survival rate for patients with or without lymph node metastasis were 77.1％ and 30.4％,respectively,with significant difference between the 2 groups (x2 =7.374,P ＜ 0.05).The 5-year survival rates of patients with malignant infiltration and inflammatory adhesion mere 51.0％ and 68.5％,with no significant difference (x2=1.148,P ＞0.05).The 5-year survival rates of patients with malignant infiltration and inflammatory adhesion in stage Ⅱ were 74.6％ and 85.7％,with no significant difference between the 2 groups (x2=0.118,P ＞ 0.05).The 5-year survival rates of patients with malignant infiltration and inflammatory adhesion in stage Ⅲ were 28.8％ and 37.5％,with no significant difference between the 2 groups (x2 =0.959,P ＞ 0.05).The results of univariate analysis showed that lymph node metastasis and TNM stage were the risk factors influencing the prognosis of patients with cT4bM0 primary rectal cancer (x2=6.468,6.596,P ＜ 0.05).The results of multivariate analysis showed that lymph node metastasis was the independent risk factor (RR =3.797,P ＜ 0.05).Conclusions En-bloc multivisceral resection should be the first surgical treatment choice for patients with cT4bM0 primary rectal cancer,and lymph node metastasis is the independent risk factor.Under the same N stage,the prognosis of patients with malignant infiltration or inflammatory adhesion is similar if R0 resection is achieved.

ObjectiveTo explore the clinical diagnosis and surgical treatment of primary retroperitonealneurilemoma(schwannoma). MethodsClinicaldataof 47patientsof primary retroperitoneal schwannoma admitted and surgically treated from January 1995 to December 2009 were retrospectivelly reviewed.ResultsAs diagnosed by pathology there were 36 cases of Benign schwannoma,with a median age at onset of 41years, among those 11 patients were symptomatic, and 25 were asymptomatic. There were 11 malignant 11 cases, the median age was 38 years, among those 6 patients were symptomatic, and 5 were asymptomatic. The positive diagnostic rate of preoperative CT and MRI were 36. 2％ ( 17/47 ) and 58. 3％ ( 7/12 ) respectively. Immunohistochemically positive rates of S-100 were 100％ and 81.8％(9/11) in benign and malignant group respectively.All cases underwent surgical treatment. Surgical resection rates for benign and malignant groups were 100％ and 90. 9％(10/11)respectively. There was no perioperative death, Overall 5-year survival rates were 100％ and 45.5％ for benign and malignant tumors groups respectively. In benign group 2 cases recurred, in malignant group 4 cases recurred, and 3 had distant metastasis.ConclusionsPrimary retroperitoneal schwannomas are less common. It is difficult to make an accurate preoperative diagnosis. Surgery is the most effective therapy.Prognosis is good for benign and poor for malignant retroperitoneal neurilemomas.

ObjectiveTo study the using of cerrobase as the compensation material in the intensitymodulated radiation therapy (IMRT) implementation and impact factors.MethodsWith therapy planning system (TPS) exported the radiation field intensity file (Dicom RT),through measuring the attenuation coefficient of cerrobase,to calculate the processing depth of AUTIMO 3D CNC corresponding for Dicom RT files at each pixel,then using the processed foam casting of Cerrobase,produced the required IMRT compensator.Through the MATRIXX testing the IMRT compensator in clinical implementation.At the same time we compared the MU of using multi-leaf collimator (MLC) and Cerrobase IMRT compensator for 10patients.ResultsWith cerrobase compensation IMRT can get similar dose or dose distribution to dose produced by TPS for point or plane dose,error is within 5％.To comparison with MLC,using cerrobase compensator has fewer treatment times ( (4.44±0.39) min:(5.71±0.57) min (t =10.82,P =0.000) )and fewer MU (462.5 ± 65.8) MU:(524.5±99.6) MU(t=3.14,P=0.012) ).Conclusions Comparison with MLC IMRT,the cerrobase compensation technique has an important application value with its unique advantages.This research provides an implemented method of IMRT radiotherapy for the primaryhospital.

Objective To establish the HepG2 cell lines which can stably express and replicate hepatitis 13 virus (HBV).Methods One point two X unit length of HBV genome was cloned intn SalⅠsite of the eukaryotic expression vector pREP10 to construct the recombinant plasmid pREP-HBV. Human hepatoblastoma cell HepG2 was transfected with pREP-HBV by Lipofectamine 2000 and seh,cted by bygromycin at the concentration of 250?g/mL.HBsAg and HBeAg were monitored by enzyme linked immunosorbent assay (ELISA)kits.H13V particles presemed in supernatant were ex- amincd by electronic microscopy.DNA isolated from intracellular HBV core particles was analyzed by Southbern blot using HBV-specific probe.Results The recombinant vector pREP HBV containing 1.2?unit length of HBV DNA was constructed successfully.After transfection of pREP-HBV to HepG2 cells and consistently cultured in hygromycin selective medium.5 drug-resistant cell lines, RHBV1-RHBV5.were established,and all of them could stably express HBsAgand HBeAg.South ern blot analysis revealed that HBV could replicate in all cell lines,as confirmed by the presence of replicateintermediatc DNA in intracellular HBV core particles.Clustered 42 nm Dane particles as well as 22-26 nm spherical H13sAg particles in condensed cuhure supernatant were visualized by elec tronic microsopic analysis.Conclusion HepG2 ceil lines in which HBV can replicate and express specific antigens are successfully established.Up to now,the cells have been passaged every three days for 50 times.

Adenoviruses harboring E. coli cytosine deaminase gene (AdCD) were used to transfect murine FBL-3 ery-throleukemia cells in vitro. FBL3 cells infected with AdCD were more sensitive to 5-fluorocytosine (5-FC) than cells infected with a control adenovirus AdLacZ. Further study indicated that this combination therapy (AdCD and 5-FC) killed tumor cells by inducing apoptosis of FBL-3 cells. The supematants from FBL-3 cells treated with AdCD/5-Fc were transferred on the culture system of uninfected (wild - type) FBL-3 cells, the result indicated that only 6.25% of the supernatant could induce significant cytotoxicity on wild type FBL3 cells. The results demonoustrated that bystander effect plays an important role in AdCD-mediated cytotoxicities. Direct injection of AdCD into established subcutaneous FBL3 tumor in mice followed by daily intraperitoneal injection of 5-FC for 10 days was found to inhibit tumor growth significant-

Antitumor effect of combined transfer of suicide gene and cytokine gene was evaluated in the present study. Adenoviruses expressing E. coli. cytosine deaminase (AdCD) and adenoviruses expressing murine interleukin 2 (AdTL2) were used for the treatment of tumor-bearing mice. The mice were inoculated s. c. with FBL-3 leukemia cells and 3 days later received intratumoral injection of AdCD in the presence or absence of AdIL2 followed by intraperitoneal 5-fluorocytosine (5FC) administration. The results demonstrated that tumor-bearing mice treated with AdCD/5FC in combination with AdTL2 showed more .potent inhibition of tumor growth and survived much longer as compared with mice treated with AdCD/5FC, AdEL2, AdlacZ/5FC or PBS. It was illustrated that the tumor mass showed obvious necrosis and inflammatory cell infiltration, and more CD4+ and CD8+ T cells infiltrated into the tumor after combined therapy. The splenic NK and CTL activities increased significantly in mice after combined transfer of CD gene and EH gene. Our results demonstrated that combined transfer of suicide gene and IL-2 gene could inhibit the growth of established tumor in mice significantly and induce antitumor immunity of the host efficiently.

OBJECTIVETo explore in vivo metabolic changes in abnormal savda patients with different types of tumor.

METHODSA total of 142 abnormal savda patients with common cancer types were enrolled in this study, and 50 healthy volunteers were recruited as the control group. For each sample, the H Nuclear Magnetic Resonance (NMR) based metabonomic analysis was performed. The free attenuation signal was computed subsection integral. Data obtained were analyzed by the Orthogonal Partial Least-Squares Discriminant Analysis (OPLS-DA).

RESULTSCompared with the control group, leucine, isoleucine, valine, histidine, phenylalanine, tyrosine, alanine, creatine, lactic acid, inositol, alpha-and beta-glucose, unsaturated lipids, very low density lipoprotein (VLDL) significantly decreased (P <0.05), while glycoprotein and carnitine significantly increased (P <0. 05) in the abnormal Savda group.

CONCLUSIONAbnormal savda patients with different types of tumor had similar metabonomics changes.

Discriminant Analysis ; Humans ; Least-Squares Analysis ; Lipids ; blood ; Magnetic Resonance Spectroscopy ; Metabolome ; physiology ; Metabolomics ; Neoplasms ; metabolism