Objective To study the relationship of expression of nuclear factor kappab gene and MMP-2mRNA and with lymph node metastasis of pancreatic cancer.Methods The expression of NF-?B was investigated by Western blot in normal pancreatic(NP) samples of 9 cases and pancreatic cancer(PC) samples of 45 cases(lymph node metastasis 30 cases,non-lymph node metastasis 15 cases).The expression of MMP-2mRNA was detected through semiquantitative reverse transcriptase polymerase chain reaction(RT-PCR).Results Of 45 PC cases,the positive expression rate of p65 and MMP-2mRNA was 66.7%(30/45) and(57.7%)(26/45),respectively.Of 30 cases of lymph node metastasis,the positive expression rate of p65 and MMP-2mRNA was 83.3%(25/30)and 73.3 %(22/30),respectively.Of 15 cases of non-lymph node metastasis,the positive expression rate of p65 and MMP-2mRNA is 33.3%(5/15) and 26.7%(4/15),respectively,and the expression of p65 gene was positively correlated with MMP-2mRNA(expression)(r=0.743,P

The early diagnosis of serious infection is very important since the fatality rate is quite high.Traditional diagnostic methods of infection includes procalcitonin,leukocyte count,CRP and so on,of which the sensitivity and specificity is not able to achieve the early diagnosis.Recently,neutrophil CD64 has been widely concerned for the high sensitivity and specificity in early diagnosis of bacterial infection.Flow cytometry is applied to detect neutrophil CD64 in early diagnosis of infectious diseases including respiratory infection,septicemia,neonatal intensive infection,burn and postoperative infection.What's more,the sensitivity and specificity can be further improved if neutrophil CD64 was combined with other inflammatory markers.Thus,neutrophil CD64 detected by flowy cytometry plays an important role in the diagnosis,monitoring,prognosis,therapeutic effect evaluation of infectious diseases.

Background To construct and analyze the differentially expressed gene library in the SD rat's brain of Parkinson's disease.Methods The lateralization Parkinson's disease model of SD rat was established by stereotaxis injection of 6-OHDA in lateralization corpora striatum. Suppression subtractive hybridization was utilized to isolate the cDNA fragments in both control and experimental groups. Then the cDNA fragments were directly bound to pMD18-T to set up the subtractive library. Amplification of the library was carried out by transformation of E. coli with high voltage electroperforation. Ninety five positive bacteria clones were randomly picked up and identified by colony PCR.Results The amplified library contained more than 1500 positive bacteria clones. Ninety-five clones were analyzed randomly by colony PCR. And there were 81% clones contained 100 ～600 bp DNA fragments. They might be the cDNA fragments of differentially expressed genes of Parkinson's disease.Conclusions A differentially expressed genes subtracted cDNA library of Parkinson's disease of SD rat was constructed successfully with SSH and T/A cloning techniques. The library is efficient and it was useful in screening and cloning new and specific differential expression genes of Parkinson's disease.

Objective:To investigate whether PUMA gene transfection can increase sensitivity of pancreatic cancer cells (PC) to 5-FU-induced apoptosis. Methods: PUMA-pCEP4 containing full length PUMA cDNA or pCEP4 was transfected into human pancreatic cancer cell line AsPC-1 by lipofectamine transfection, G418 selection was used to select positive cells. AsPC-1, AsPC-1/PUMA and AsPC-1/pCEP4 cells were separately treated with serial concentrations of 5-FU(0.01-100 ?mol/L). MTT assay was used to determine the cell survival rate in each group and IC50 of 5-FU was calculated. TUNEL,FCM and DNA ladder observation were employed to study cell apoptosis. Western blotting was performed to detect the expression of PUMA protein. Results: The 5-FU IC50 values of AsPC-1, AsPC-1/PUMA and AsPC-1/pCEP4 cells were (12?1.9)?mol/L,(1.6?0.4)?mol/L and (10.4?1.6) ?mol/L, respectively, with the sensitivity of AsPC-1/PUMA cells increased by 7.5 folds. 5-FU induced cell apoptosis of AsPC-1 cells in a dose-dependent manner, with the apoptosis of AsPC-1/PUMA cells more prominent than those of AsPC-1 and AsPC-1/pCEP4 cells. Low concentration of 5-FU (0.1 ?mol/L) induced few apoptosis of AsPC-1/pCEP4 cells([1.14?0.28]%) and AsPC-1 cells ([0.9?0.23]%), and induced apoptosis in AsPC-1/PUMA cells([6.47?1.42]%). High concentration of 5-FU (1.0 ?mol/L) induced apoptosis in all groups, with that in AsPC-1/PUMA cells([34.54?9.36]%) significantly higher than those in AsPC-1/pCEP4 cells([15.8?5.15]%) and AsPC-1 cells ([12.8?3.74]%, both P

objective To construct the recombinant adenovirus containing p53 up-regulated modulator of apoptosis(Ad-PUMA)and investigate its growth inhibition effect on pancreatic callCer cells in vitro and in vivo.Methodls Ad-Easy system was used to construct Ad-PUMA by recombination in E.coli.The virus was Dackaged in 293 cells and subsequently identified valid.The AsPC-1 cells were infected with AdPUMA.Before and after Ad-PUMA infection,the expression of PUMA protein wag investigated by western blot,the inhibition rate of AsPC-1 cells was examined by MTY assay.The in vivo tumor suppressive effect was detected in nude mice with human AsPC-1 xenograft.PUMA protein and the apoptosis of AsPC-1 xenograft were detected by western blot and TUNEL(terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling)method.Results In vitro,the expression of PUMA protein was increased with titer of Ad-PUMA,the proliferation of AsPC.1 cells were suppressed,significantly,and the effect was in a viral dose-dependent nlanner.In vivo,the growth in nude mice of AsPC-1 infected with Ad-PUMA was significantly inhibited with an inhibition rate of 44.2%.The expression of PUMA was significantly up-regulated,and the apoptosis index wa8 significantiv increased in tumor after Ad-PUMA infection as determined by western blot and TUNEL.Conclusion The expression of PUMA call inhibit the proliferation of pancreatic cancer in vitro and in vivo,and may be used 88 a potential tool for cancer therapy

Background and purpose:Members of the NF-kappaB family of transcription factors could activate bcl-2 anti-apoptotic genes at the transcriptional level and then regulate cell apoptosis.The p53 tumor suppressor gene has a clear role!in both the regulation of cell cycle and expression of apoptosis-related bcl-2 family.There were few reports about the expression of NF-?Bp65,bcl-2,bcl-xL and their relationships with p53 and cell apoptosis in pancreatic cancer.The present study was designed to analyze the expression of the antiapoptotic molecules nuclear factor kappaB(NF-?Bp65),bcl-2 and bcl-xL in pancreatic ductal carcinoma(PC) and their correlations to the apoptosis index and p53 expression.Methods:Immunohistochemical analyses of P53 protein was performed on 25 cases of pancreatic ductal carcinoma and 9 cases of normal pancreas;Western blot was used to evaluate NF-?Bp65 protein and RT-PCR for bcl-2 mRNA and bcl-xL mRNA of 25 cases of pancreatic ductal carcinoma and 9 NP cases.The apoptosis was determined by TUNEL.Results:The positive rate of P53 protein was 56%(14/25)in PC tissues,significantly higher than that in NP tissues(P

Objective To study the intelligence and memory impairment of patients with craniocerebral injury during traffic accident and its correlated factors. Method Data of psychological tests, eletroencepa-lograph (EEG) and brain image information of 652 subjects with brain injury who survived from traffic accident in Chengdu from Jan. 1,1999 to Dec. 31,2001 were collected and analysed. Results The average intelligence quotient (IQ) of patients in patients group was 77.85?15.97. 27.9 % patients's IQ was lower than 70. 51.9% patients's memory quotient (MQ) was lower than 51. The intelligence impairment was correlated with education level, left temporal lobe injury, three or more lobes of the brain injuried and EEG abnormality. Memory impairment was correlated with education level, interval between injury and e-valuation, brain stem injury, left temporal lobe injury, and EEG abnormality. Conclusion Impairment of intelligence and memory in different degrees were observed in most patients with traumatic brain injury during traffic accident. Intelligence and memory impairment were correlated with left temporal lobe injury, EEG abnormality, brain stem injury and education level.

Objective To investigate the activation of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated protein kinase (ERK) signaling pathway in the endometrium of women with polycystic ovary syndrome (PCOS) and its effect and significance in the cause of hyperplasia and carcinoma;and investigate the factors which affect the activation of the MAPK/ERK signaling pathway. Methods Collected 52 patients diagnosed as PCOS who were taken dilation and curettage of uterus as study, while 32 non-PCOS patients matched as control group. Serum hormonal parameters, fasting blood glucose and insulin were measured in all patients. The PCOS patients were sub-group as insulin resistance group and non-insulin resistance group; all the patients were carried out pathology inspection of endometria, and the PCOS patients were sub-group as endometrial hyperplasia and carcinoma group and normal endometrium group based on the outcome of pathology inspection. Western blot were performed to detect the expressions of ERK1/2 and phosphorylated ERK1/2 (p-ERK1/2), the activation of ERK1/2. Results (1)The expression of pERK1/2 [(61 ±13)%] in the endometrium in PCOS group was higher than that in the control [(44 ±10)%, P ＜0.01]. (2)The expression of p-ERK1/2 was significantly increased in group of endometrial hyperplasia and carcinoma [ ( 70 ± 11 )% ] compared to that in group of normal endometrium [ (55 ± 10)% ,P ＜ 0.01 ], while there were significant difference between group of insulin resistance [ (63 ± 13 )% ] and group of non-insulin resistance [ (55 ±7)%, P ＜0.01 ]. (3) Fasting insulin level, insulin area under the curve and body mass index were related to the expression of p-ERK1/2 in endometrium with PCOS, the correlation coefficient were 0.447, 0.456 and 0.381, respectively ( all P ＜ 0.01 ). Conclusions The MAPK/ERK signaling pathway in endometrium with PCOS was overactivation, which was related to the endometrial hyperplasia and carcinoma; while the activation of MAPK/ERK signaling pathway were effected by insulin resistance and hyperinsulinemia.

Objective To investigate the expression changes of nuclear factor kappa B (NF-?B) and matrix metalloproteinase-9 (MMP-9) in the cultured hepatocellular carcinoma cells 9204 (HCC9204) transfected with inhibitory kappa B alpha(I?B-?)vector. Methods After pcDNA3-I?B-? vector and pcDNA3 were transfected into HCC9204 by lipofectamine method, Western-blot and RT-PCR analysis were used to detect the expressions of NF-?B and MMP-9. Migration and invasion of tumor cells were assayed by fundus membrane invaded by them. Results When pcDNA3-I?B-? was transfected into HCC9204, the expression of NF-?B was decreased at the protein level, and the expression of MMP-9 mRNA and the invision and metastasis ability of transfected cells were obviously decreased. Conclusion When the activity of NF-?B is inhibited, the ability of invasion and metastasis in HCC9204 cells decrease, which could be related to the decreased the expression of MMP-9.

0.05), but there were significant differences after the operation (P