Objective To observe the process of Vibrio vulnificus inducing dendritic cell strain apoptosis.Methods We established the mixed culture model of mouse dendritic cell ( DC 2.4 strain) and Vibrio vulnificus( Vv1.1758 strain ),analyzed morphological characteristics of cell apoptosis by DAPI fluorescence staining,detect DNA fragmentation level of apoptosis cells by DNA Ladder assay,analyze DC2.4 apoptosis rate by Annexin V FITC/PI staining,determine activities of caspase-3 and caspase-8 by means of spectrophotometric method and detect changes of mitochondrial transmembrane potential ( △ Ψm ) by JC-1 fluorescence labeling.Results After Vv1.1758 strain and DC2.4 cell were mixed and cultured for 4 h,DAPI fluorescence staining showed typical apoptosis characteristics-chromatin condensation and marginalization; DNA agarose gel electrophoresis showed apoptosis band; apoptosis rates at 2,4 and 6 h were respectively (37.8±9.8) ％,(54.3 ± 12.7 ) ％ and ( 68.2± 14.6 ) ％ ; Mitochondrial transmembrane potentials (△Ψm) at 1 h,2 h and 4 h reduced by 7.1％,16.1％ and 46.7％ respectively; caspase-8 activity increased at 1.5 h and reached the peak at 2 h [ (2.48±0.19) U/μg],while caspase-3 activity started to increase at 3h and reached the peak at 4 h [ ( 1.91 ±0.16) U/μg ].Conclusion Vibrio vulnificus could induce dendritic cells by two pathways: reducing mitochondrial transmembrane potential and activating caspase-8 promoter and finally activate effector caspase-3 to promote apoptosis.

Objective To investigate the invasion,internalization and the organelles damage of the cultured dendritic cells ( DC2.4 strain) during Vibrio vulnificus (Vv) infection.Methods The study model was the cultured DCs infected by Vv 1.1758 strain.Electron microscopy was used to observe the localization of bacteria in different time point of infection,cell morphology and the process of organelles changes.The cytoskeleton structure including the microfilaments and the microtubules rearrangement was examined by the fluorescence microscope.Results The Vv were pinocytosed into the DC cells through double-sides,and localized at 1-2 μm of the inner side membrane.It cost 1.27,1.87,3.43 hours reaching the infection ratio of 25％,50％,75％,respectively.Using electron microscopy,the DCs had been observed the phagosome formation within 1h,chromatin activation within 2 h,chromatin aggregation 4 h,and the significant cytoskeleton structure disruption within 6 h.Endoplasmic reticulum,mitochondria and lysosomes became swollen.In DCs,the protruding filaments gradually reduced,and their shape changed from the point-like to the linearlike aggregation at the inner side of the plasma membrane,extended microtubules disappeared,the microtubules at the outside nuclear membrane striking rearranged.Conclusion After DC was infected by Vv,the bacteria were pinocytosed into the inner side of DC membrane,and the microfilaments were observed to move from the cytoplasm to cell membrane.In addition,the microtubules moved from the synapse and the cell membrane to the nuclear membrane.The high lethality of Vv could provoke to the DCs cytoskeleton rearrangements.

Objective To analyze the correlation between neutrophil-to-lymphocyte ratio(NLR),platelet-to-lymphocyte ratio(PLR) in peripheral blood and the therapeutic effect and prognosis of patients with advanced lung adenocarcinoma treated with tyrosine-kinase inhibitors.Methods Retrospective analysis was utilized to measure the clinical pathological characteristics and follow-up data of 40 cases with advanced lung adenocarcinoma definitely diagnosed by pathology.Correlation analysis was performed by Spearman test,and comparison of the sample rate was measured by the chi square test.Survival analysis was performed by Kaplan-Meier method,Log-rank test was used to compare patients survival difference among different groups.COX regression model was utilized to analyze the prognostic factors of patients with lung cancer.Results Through the ROC curve,the optimal cut-off points of NLR and PLR were calculated to be 3.74 and 205.1 respectively.In NLR<3.74 group,objective response rate(ORR),the median progression-free survival(PFS) and the median overall survival(OS) were 86.9%(20/23),9.02 months and 12.37 months respectively.In NLR ≥3.74 group,ORR,the median PFS and the median OS was ORR were 35.2%(6/17),3.15 months and 5.12 months respectively.Compared with NLR ≥ 3.74 group,NLR<3.74 group had a higher ORR(P=0.036),PFS(P=0.011) and OS(P=0.021),the difference was statistically significant.In PLR<205.1 group,ORR,the median PFS and the median OS was ORR were 85.7%(18/21),9.67 months and 14.35 months respectively.In PLR ≥ 205.1 group,ORR,the median PFS and the median OS was ORR were 42.1%(8/19),3.21 months and 9.25 months respectively.Compared with group PLR ≥ 205.1,PLR<205.1 group had a higher ORR(P=0.045) and PFS(P=0.018),the difference was statistically significant.COX multivariate analysis showed that NLR was an independent prognostic factor for PFS(RR=1.091,95%CI:1.004-1.185,P=0.039) and OS(RR=0.986,95%CI:0.932-1.148,P=0.021).PLR was an independent prognostic indicator of PFS(RR=1.100,95%CI:0.996-1.005,P=0.024).Conclusion NLR and PLR may be independent prognostic factors in patients with advanced lung adenocarcinoma.

OBJECTIVETo explore the prognostic factors for rectal cancer patients with synchronous liver metastases.

METHODSData from a total of 77 cases of rectal cancer with synchronous liver metastases treated in our center from January 2002 to December 2008 were collected and reviewed. The total survival rate was analyzed by Kaplan-Meier method. Log-rank test and Cox regression model with SPSS 17.0 software were used to analyze 13 factors including clinicopathological factors and treatment choices.

RESULTSThe median survival time of the 77 cases was 12 months. The 1-, 2-, 3- and 5-year survival rates were 47.7%, 28.0%, 13.1%, and 1.5%, respectively. Univariate analysis with Kaplan-Meier method revealed that the differentiation of the primary tumor, T-stage, N status, the distribution, number and size of liver metastases, extrahepatic disease, serum CEA level at diagnosis and treatment modality were prognostic factors (P < 0.05). Multivariate analysis showed that the differentiation of the primary tumor (P = 0.007), T-stage (P = 0.027), the size of liver metastases (P = 0.003), serum CEA value at diagnosis (P = 0.000) were independent prognostic factors for rectal cancer patients with synchronous liver metastases.

CONCLUSIONThe factors affecting the prognosis for rectal cancer patients with synchronous liver metastases are the differentiation of the primary tumor, T-stage, N status, the distribution, number and size of liver metastases, extrahepatic disease, serum CEA level at diagnosis and treatment modality. The differentiation of the primary tumor, T-stage, the size of liver metastases, and serum CEA value at diagnosis are independent prognostic factors.

Adult ; Aged ; Aged, 80 and over ; Carcinoembryonic Antigen ; blood ; Female ; Follow-Up Studies ; Humans ; Liver Neoplasms ; pathology ; secondary ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Proportional Hazards Models ; Rectal Neoplasms ; blood ; drug therapy ; pathology ; surgery ; Retrospective Studies ; Survival Rate

OBJECTIVETo clone and express the Rv3871 gene related to the virulent protein secretion of Mycobacterium tuberculosis and analyze its molecular structure, function and homology using bioinformatic approach.

METHODSA pair of primers was designed to amplify the Rv3871 gene, which was subcloned into the prokaryotic plasmid pET32a(+). The recombinant plasmid was identified by sequence analysis and the expressed recombinant protein by SDS-PAGE. The structure, function and homology alignment of Rv3871 were analyzed comparatively against other mycobacteria.

RESULTSThe restriction fragments through molecular cloning matched perfectly in size with our prediction. The gene sequence was consistent with the corresponding sequence in GenBank. The expression protein was detected by SDS-PAGE with a molecular weight of 84 kD. Two FtsK/SpoE III domains were found by bioinformatic analysis. The homology results showed distinct differences between Rv3871 of the pathogenic M. tuberculosis and its counterparts in non-pathogenic mycobacteria.

CONCLUSIONMolecular cloning, expression and sequencing identify the structural and functional characteristics of Rv3871. The structural and functional differences of the gene between pathogenic and non-pathogenic mycobacteria identified by bioinformatics provide some evidence for the pathogenesis and drug targets of tuberculosis.

Bacterial Proteins ; genetics ; metabolism ; Cloning, Molecular ; Computational Biology ; Mycobacterium tuberculosis ; genetics ; metabolism ; pathogenicity ; Recombinant Proteins ; genetics ; metabolism ; Virulence ; genetics

GATA transcription factor family members have been found to involve in the growth and differentiation of mammary gland. Among them GATA-3 is regarded as the most critical regulator involving the tumorigenesis of breast cancer (BC). Recently, trichorhinophalangeal syndrome-1 gene (TRPS-1), a new GATA family member, has been identified to be highly prevalent in breast cancer. Compared with ER-negative breast cancer, the expression of TRPS-1 is higher in ER-positive breast cancer and was significantly correlates with estrogen receptor, progesterone receptor, and GATA-3, indicating it may serve as a ductal epithelial cell-specific regulator in the differentiation of breast ductal epithelial cells. Studies have shown that miR221/222 is able to downregulate the expression of an epithelial cell marker E-cadherin by targeting TRPS-1, resulting in mammary epithelial cells transition to mesenchymal cell (EMT). In addition, it has been well accepted that, and the Science and Technology Bureau of Jiaxing (2012AY1071-2)TRPS-1 plays a role in the differentiation of several other cell types including kidney nephric mesenchymal cells, columnar chondrocytes, and osteoclasts, indicating that TRPS-1 involves in mesenchymal-to-epithelial cell transition (MET). In this article, we summarize the roles of GATA transcription factor TRPS-1 in ductal epithelial cells and the roles of its gene and protein expressions in predicting the prognosis of breast cancer.
Breast Neoplasms ; genetics ; pathology ; DNA-Binding Proteins ; genetics ; Epithelial-Mesenchymal Transition ; Female ; GATA3 Transcription Factor ; genetics ; Humans ; Transcription Factors ; genetics

Porcine epidemic diarrhea virus (PEDV) causes lethal diarrhea in piglets that leads to great economic losses in East Asia. It was reported that aminopeptidase N (APN) was the receptor for Transmissible gastroenteritis virus (TGEV), Human coronavirus 229E (HCoV-229E) and Feline coronavirus (FeCoV) which all belonged to group I coronavirus including PEDV. It was also confirmed previously that porcine aminopeptidase N (pAPN) could bind to PEDV, and anti-pAPN antibodies could inhibit the combination. To investigate whether pAPN was a receptor for PEDV, we transfected MDCK cells with porcine aminopeptidase (pAPN) cDNA and this enabled non-susceptible cells to support PEDV replication and serial viral propagation. Moreover, the infection was blocked by antibodies against pAPN, implying the critical role of pAPN during virus entry. In addition, immunofluorescence assays for detection of pAPN and PEDV antigens, together with neutralization assays using antibodies against pAPN, further confirmed the correlation between pAPN expression and viral replication in pAPN-transfected MDCK cells. These results indicated that pAPN is a functional receptor for PEDV.
Animals ; Antibodies ; pharmacology ; CD13 Antigens ; antagonists & inhibitors ; genetics ; metabolism ; Cell Line ; Coronavirus Infections ; enzymology ; metabolism ; Dogs ; Porcine epidemic diarrhea virus ; enzymology ; Swine

OBJECTIVETo observe the effect of targeted argon-helium cryoablation on portal region of the liver in dogs by observing the pathological changes in the first-order branches of the Glisson ductal system.

METHODSTwelve healthy dogs underwent percutaneous targeted argon-helium cryoablation of the liver and sacrificed at 3 and 28 days after the cryoablation to observe the pathological changes in target area for cryoablation and the first-order branches of the Glisson ductal system.

RESULTSNo obvious damage was not found in the vascular wall of the portal vein by gross or microscopic observation, but the liver tissue in the vicinity of the blood vessels showed total necrosis. In spite of the injuries of different degrees in the first-order bile duct system after argon-helium cryoablation, no severe damages such as perforation or full-thickness necrosis occurred in bile duct wall, and most of the injuries were temporary and reversible. The size of the ablated area on day 28 was significantly reduced as compared with that on day 3 following the cryoablation (P<0.05). In the acute stage after the cryoablation (1-3 days), ALT and AST levels increased significantly in (P<0.05) but recovered 1-4 weeks later (P>0.05). The cryoablated area was basically consistent with the pathological area that underwent necrosis (P>0.05).

CONCLUSIONTargeted argon-helium cryoablation can cause total destruction of the liver tissue around the blood vessel without damaging the vascular walls of the portal vein. Argon-helium cryoablation induces relatively minor injuries to the bile duct of hepatic portal section and does not obviously damage the liver function, and the scope of tissue necrosis can be estimated according to the size of frozen area observed. Argon-helium cryoablation is a safe and minimally invasive operation with reliable therapeutic effect.

Animals ; Argon ; Bile Ducts, Extrahepatic ; pathology ; Cryosurgery ; adverse effects ; methods ; Dogs ; Female ; Helium ; Liver Neoplasms, Experimental ; surgery ; Male ; Portal Vein ; pathology ; Random Allocation

OBJECTIVETo summarize the clinical application results of the micro-decompression procedure for the treatment of lumbar spinal stenosis with multilevel.

METHODSFrom January 2004 to December 2008, 40 patients with lumbar spinal stenosis with multilevel were treated by micro-decompression procedure. There were 28 males and 12 females, ranging in ages from 55 to 80 years,with an average of 58 years. The course of this disease ranged from 18 months to 12 years,averaged 32 months. Forty patients with lumbar spinal stenosis with multilevel were diagnosed by CT or MRI examination. There were 20 cases with two levels stenosis (L4,5 and L5S1), 15 cases with three levels stenosis (L3,4, L4,5 and L5S1), and 5 cases with four levels stenosis (L2,3, L3,4, L4,5 and L5S1). The therapeutic effects were evaluated according MacNab standard in aspect of pain, bladder function, range of lumbar spine motion and muscle strength of lower limb.

RESULTSAll of the incisions healed without infections and complications. The mean operation time of each side was 70 minutes (ranged from 50 to 90 minutes), and mean blood loss was 150 ml (ranged from 90 to 200 ml). All the patients were followed up from 22 to 52 months with an average of 26 months. According to MacNab standard, 28 cases got an excellent result, 10 good and 2 poor.

CONCLUSIONOperative treatment for lumbar spinal stenosis with multilevel should focus on the symptom sites causing by neural compression and preventive decompression operations are not necessary for nonsymptomatic sites. The micro-decompression procedure can be easily tolerated by older patients;it can decrease the damage to the posterior stabilizing structures of the lumbar spine. It is easily to access to spinal canal and decompress the nerve roots.

Aged ; Aged, 80 and over ; Decompression, Surgical ; methods ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Spinal Stenosis ; surgery

Objective:To investigate the killing effect of radiation on echinococcus in vitro culture and its effect on the mRNA expression of growth arrest and DNA damage 45 alpha (Gadd45α) gene. Methods:Echinococcus from naturally infected sheep liver was cultured in vitro and divided into 7 groups. The echinococcus was irradiated with 6 MeV at doses of 0 (control), 20, 40, 60, 80, 100, and 120 Gy, respectively. The growth of echinococcus was observed under light microscope at 1, 3, 5 and 7 d after the radiation. The expression of Gadd45α mRNA in control, 20, 40 and 60 Gy groups of echinococcus was detected by RT-PCR technique at 7 d after the radiation. Results:The disintegration and exfoliation of echinococcus were observed under light microscope at 1, 3, 5 and 7 d after the radiation, and the death rate of echinococcus was positively correlated with the radiation dosages ( r = 0.81, P < 0.05). After the radiation at 7 d, compared with the control group (100.00 ± 0.00), the mRNA expression levels of Gadd45α in echinococcus of 20, 40, and 60 Gy groups were significantly increased (279.74 ± 80.08, 759.38 ± 160.98, 1 666.68 ± 316.36, P < 0.01), and it was positively correlated with the radiation dosages ( r = 0.93, P < 0.01). Conclusion:Radiation has a certain killing effect on echinococcus cultured in vitro, and there is a certain dose-effect relationship with the radiation dosages, and Gadd45α gene may be involved in the molecular mechanism of radiation-induced killing of echinococcus in vitro.