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

Objective To explore the relationship between injury of corticospinal tract (CST), diffusion anisotropic features of white matter fibers and cerebral infarction prognosis in patients with different ages by MRI and diffusion tensor imaging (DTI). Methods 24 cases of cerebral infarction were divided into 3 groups according to age: Group A (n=8, 45～59 years old), Group B (n=8, 60～74 years old) and Group C (n=8, 75～82 years old). DTI were acquired in the patients on a 3.0 T MR imaging scanner in the first 3 days. Before and 6 weeks af-ter treatment, Fractional anistrophy (FA) value of the infarcted focus and the contralateral corresponding brain tissues were measured, and the nerve function were measured by National Institutes of Health Stroke Scale (NIHSS). Results FA values of the infarcted focus were less than the FA of contralateral corresponding brain tissues (P<0.05) and they were significantly improved 6 weeks after treatment (P<0.05).The NISHH scores of corticospinal tract (CST) 2 were higher than that of CST 1 in each group (P<0.05). The NISHH scores after admission were lower than the scores on admission in the same group and the same CST level. Conclusion DTT can show the relationship between the direction and morphology of the fiber tract and the infarct lesions, which can be used to evaluate the impairment of CST at the early stage of ischemic stroke, favoring the prognostic judgment in the rehabilitative therapy of cerebral infarction.

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 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:Intervertebral disc degeneration is the pathological basis of degenerative spinal diseases. Studies on the influentialfactors of intervertebral disc degeneration contribute to the prevention and treatment of degenerative spinal disease.
OBJECTIVE:To observe the growth and proliferation of nucleus pulposus cels isolated by trypsin plus type II colagenase digestion in complete medium with different glucose concentrations, exploring the optimal glucose concentration for growth of nucleus pulposus cels.
METHODS:Nucleus pulposus cels isolated and cultured by trypsin plus type II colagenase digestion method were observed under an inverted microscope, and thecelnumber was counted. Morphology of nucleus pulposus cels was observed afterhematoxylin-eosinstaining and toluidine blue staining. Colagen type II immunoreactivity was detected by immunohistochemical staining combined with immunofluorescent staining.Nucleus pulposuscels were incubated in complete medium containing various glucose concentrations (0, 6.25, 12.5, 17.5, and 25 mmol/L) for 24 hours, and then cel proliferation and apoptosis were determined.
RESULTS AND CONCLUSION:The stained nucleus pulposus cels showed polygonal and short spindle, with one or two nuclei. Celularpseudopod appeared gradualy and then became slim with increased passage numbers. The isolated and cultured nucleus pulposus cels positively expressed colagen type II and aggrecanProliferative activity of nucleus pulposus cels cultured in medium with 17.5 mmol/L glucose was significantly higher than that in medium with 0 and 25 mmol/L glucose (P< 0.05 orP< 0.01). There wasno significant differencein cel apoptosis between these groups except for 0 mmol/L glucose (P<0.05). These results confirm that a large number of nucleus pulposus celscan beharvested by trypsin plustype II colagenase digestion and the optimal glucose concentration is 17.5 mmol/L.

Objective To study The effect of a early systemic psycho-rehabilitation intervention on patients with post-stroke depression.Methods 80 patients with post-stroke depression were randomly divided into test group (n=40) and control group (n=40) by Hamilton's Depression Scale (HAMD).They were treated conventionally by neurologist. The patients in test group were added systemic psychological intervention administered by psychologist and movement training administered by professional physiatrician. And then the scores to HAMD and CNFDS (clinical neurological functional defect scoring) between two groups were evaluated until 2 weeks or 4 weeks later. Results Nerve function of the both groups was improved,and score to HAMD in the test group was less than that in control group (P

Objective To prepare highly expressed, purified and refolded SCR15-18 of human soluble complement receptor type 1 (sCR1-SCR15-18) protein. Methods The expression of recombinant pET32-sCR1-SCR15-18 in E.coli.. BL21 was induced by IPTG of different concentrations for different time period under different temperatures and the bacteria were split by sonication. The sCR1-SCR15-18 protein was purified by Ni2+-NTA resin affinity chromatography. The purified protein was refolded under different conditions. Then the bioactivity of the protein was analyzed. Results The sCR1-SCR15-18 protein of high expression, purity and bioactivity was attained. Conclusion The parameters of expression, purification and refolding of sCR1-SCR15-18 protein were optimized, which may pave a way for further studies.

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 study the expression of reversion-inducing-cysteine-rich protein with Kazal motifs (RECK) in human pancreatic carcinoma tissues and pancreatic carcinoma cell lines; the effects of recombinant lentiviruses carrying RECK gene(LV-RECK) therapy on human pancreatic carcinoma xenograft in nude mice; and to find out the relationship between the expression of RECK and the prognosis of pancreatic carcinoma.Methods Immunohistochemical method was used to detect the expression of RECK in the resected specimens of pancreatic carcinoma and their corresponding normal pancreatic tissues in 42 patients.Western blotting was used to examine the expression of RECK in human pancreatic carcinoma cell lines (PANC-1,MIAPaCa-2,AsPC-1).Statistical analyses were performed to determine the relationship between RECK expression and the clinicopathological characteristics and prognosis in pancreatic carcinoma.Subcutaneous xenograft tumor models of human pancreatic carcinoma were established in nude mice.These nude mice were then divided into the experimental group,the negative control group and the blank control group randomly.The three groups of nude mice were intratumorally injected with LV-RECK,LV-EGFP and normal saline (NS) respectively.The antitumor effect was studied.Immunohistochemical method was used to detect the expression of RECK and microvessel density (MVD).Terminal deoxynucleotidyl transferase mediated dUTP-DIG nick end labeling (TUNEL) was used to detect the apoptosis of tumor cells.Survival analysis was performed.Results All three pancreatic carcinoma cell lines did not express RECK.The overall positive rate of RECK expression was 45.2 ％ (19/42) in pancreatic carcinoma,and 88.1 ％ (37/42) in normal pancreatic tissue.The expression level of RECK was significantly lower in the tumor tissues than in the normal tissues (P＜0.01).The expression of RECK was significantly associated with TNM stage,lymph node metastasis and local infiltration of pancreatic carcinoma (P＜0.05).Kaplan-Meier survival analysis revealed that the survival time was significantly longer in the RECK positive patient group than in the RECK negative patient group.Univariate Cox regression analysis revealed that RECK expression,TNM stage,lymph node metastasis and local infiltration were significantly related with prognosis for pancreatic carcinoma (P＜0.05).Multivariate Cox regression analysis revealed that only RECK expression remained as an independent significant factor in predicting the prognosis of pancreatic carcinoma (P ＜ 0.001).When compared with the negative control and the blank control groups,the volume of subcutaneous xenograft tumor in the experimental group was significantly decreased (P＜0.05).RECK protein in the experimental group was re-expressed.MVD of the experimental group was significantly less than those of the control groups (P＜0.05).Apoptotic index (AI)of the experimental group was significantly higher than those of the control groups (P＜0.05).The survival time of nude mice in the experimental group was significantly longer than those in the control groups (P＜0.05).Conclusions RECK expression was closely related to invasion,metastasis and prognosis of pancreatic carcinoma and it was an independent prognostic marker.RECK gene over-expression inhibited neovascularization of pancreatic carcinoma,induced apoptosis of tumor cells,inhibited the growth of tumor xenograft and improved the prognosis of tumor-bearing mice.These results suggest a possible new treatment for pancreatic carcinoma.

BACKGROUND:Artificial lumbar disc replacement is a new choice for the treatment of degenerative disc disease, and preserves lumbar vertebra’s biomechanical characteristics during pain elimination. The design of the prosthesis structure and material needs further study and validation. OBJECTIVE:To review the structure and material types of presently designed artificial lumbar discs, then to discuss the trends in the optimization design of prosthesis. METHODS:The PubMed database, China National Knowledge Infrastructure database and Chinese BioMedical Literature Database were searched for related articles concerning artificial lumbar disc and type and biomechanics of nucleus pulposus prosthesis material published from January 2005 to February 2013 by the first author. Key words were“artificial lumbar disc, principle of prosthesis design, structure, material, clinical trials”in Chinese and“artificial lumbar disc, total disc replacement, structure, material, clinical trial”in English. Repetitive and old studies were excluded. 135 articles were found, but 36 articles were included for review. RESULTS AND CONCLUSION:At present, the materials for intervertebral discs include cobalt-chromium al oy, ceramics, stainless steel, titanium al oy and ultrahigh molecular weight polyethylene. Artificial lumbar disc is commonly made by different materials. Bryan prosthesis is most commonly used in the clinic. Three-dimensional finite element analysis, in vitro trial and clinical studies verified its good biomechanical property. The successful rate of replacement was high. Nucleus prosthesis contains prefabricated type and situ polymerization type, and obtains smal injury, so it is a hot focus in present study, but it cannot achieve biomechanical function of human nucleus pulposus. To dig novel material is a future direction for designing individual prosthesis. The prosthetic structure and biomaterial design experience constant improvement and development. This study combines latest study trend and prospects the development of biomimetic design, material improvement, the optimization design of prosthesis and assisted devices.