Streptococcus pneumoniae(S.pn) is an opportunity pathogenic bacteria,environmental factors play a key role in the pathogenicity of S.pn. It is important to study virulent gene in vivo. The S.pn suicide plasmid containing gfp reporter was constructed by fusing the genes pneumolysin and gfp,in which gfp is an excellent molecule probe in vivo. The plasmid was integrated to No.22 S.pn by homologous recombination. The recombinant S.pn was gained and evaluated in aspects of fluorescence excitation, biological character and physio-activity. The results showed it is efficient and available to report the expression of virulent genes in vivo and in vitro, which will provide a new easy method for evaluating and screening the virulent genes of S.pn in vivo.

PURPOSE: The aim of this study was to establish an in vitro method to purify human multipotent adult progenitor cells (hMAPCs) and assess their possible differentiation into hepatocytes by coculture with human hepatocyte line L02. METHODS: hMAPCs were isolated by magnetic activated cell sorting (MACS) depletion selection using CD45 and GlyA microbeads. After indirect or direct coculture of hMAPCs and human hepatocyte line L02, the expression of albumin (ALB), alpha-fetoprotein (AFP), cytokeratin (CK) 18, and CK19 by hMAPCs was detected by immunocytochemistry. RESULTS: With the MACS method, (5-10) x 10(4)/mL hMAPCs could be separated from 1 x 10(6)/mL bone marrow mononuclear cells. The purity of CD45-/GlyA- cells separated from bone marrow adherent cells was more than 98%, as determined by flow cytometry. In the coculture without cell-to-cell contact, hMAPCs expressed high AFP on day 1, and then tapered daily to low expression on day 7; ALB expression reached its peak on day 5, and remained high on day 7; CK18 was initially expressed on day 5 and was higher on day 7; CK19 was negative in all assays. In the coculture with cell-to-cell contact, ALB and CK18 were expressed by most cells while AFP appeared in only a few on day 5. CONCLUSION: hMAPCs were induced to differentiate into mature hepatocyte-like cells by coculture with a hepatocyte cell line, either with or without cell-to-cell contact, but the former seemed more effective.
Adult* ; alpha-Fetoproteins ; Bone Marrow ; Cell Differentiation ; Cell Line ; Coculture Techniques* ; Flow Cytometry ; Hepatocytes* ; Humans ; Immunohistochemistry ; Keratins ; Microspheres ; Stem Cells*

Brain Neoplasms ; surgery ; Computer Simulation ; Humans ; Imaging, Three-Dimensional ; methods ; Male ; Middle Aged ; Neurosurgical Procedures ; instrumentation ; methods ; Patient Care Planning ; Preoperative Care

BACKGROUNDThe virtual reality (VR) system can provide the neurosurgeon to intuitively interact with and manipulate the three dimensional (3-D) image similarly to manipulate a real object. It was seldom reported that the system was used in diagnosis and treatment of cerebral aneurysms. This study aimed to investigate the application of VR system in diagnosis and therapeutic planning of cerebral aneurysms.

METHODSA total of 24 cases of cerebral aneurysms were enrolled in this study from 2006 to 2008, which diagnosed by 3-D digital subtraction angiography (3D-DSA) or VR-based computed tomography angiographies (CTA). The VR system and 3D-DSA system were used to observe and measure aneurysms and the adjacent vessels. The data of observation and measurements were compared between VR image and 3D-DSA image. All the patients underwent surgical plan and simulated neurosurgical procedures in the VR system.

RESULTSThere were 28 aneurysms detected in VR system and 3D-DSA system. The VR system generated clear and vivid 3-D virtual images which clearly displayed the location and size of the aneurysms and their precise anatomical spatial relations to the parent arteries and skull. The location, size and shape of the aneurysms and their anatomical relationship with the adjacent vessels were similar between 3-D virtual image and 3D-DSA, but the spatial relationship between aneurysms and skull only been displayed by VR system. This VR system also could simulate simple surgical procedures and surgical environments.

CONCLUSIONSThe VR system can provide a highly effective way to provide precise imaging details as same as 3D-DSA system and assist the diagnosis of cerebral aneurysms with virtual 3-D data based on CTA. It significantly enhances the chosen therapeutic strategy of cerebral aneurysms.

Angiography, Digital Subtraction ; Humans ; Imaging, Three-Dimensional ; Intracranial Aneurysm ; diagnosis ; diagnostic imaging ; Tomography, X-Ray Computed

Cervical Atlas ; blood supply ; Humans ; Male ; Middle Aged ; Vertebral Artery ; abnormalities

Objective To investigate the change of the basic fibroblast growth factor(bFGF) leve in human detrusor muscle(DM)in bladder outlet obstruction(BOO)due to benign prostatic hyperplasia(BPH)and its implication.Methods Fifty-four patients with BPH were divided into two groups:the obstructive DM stability and instability groups;and 15 men with bladder tumor who underwent operation in the same period were enrolled in the control group.The bFGF mRNA level in DM was measured by reverse transcription and polymerase chain reaction(RT-PCR)and the bFGF protein level was measured by immunohistochemical staining method.Results The bFGF-mRNA expression level of bladder smooth muscle cells was significantly lower in the control group than that in the obstructive DM stability and instability groups(all P＜0.05),but there was no significant difference between the obstructive DM stability and instability groups(P＞0.05). Conclusions The expression level of bFGF mRNA in bladder DM is elevated in BOO due to BPH,but there is little or no correlation between the increased expression of bFGF mRNA and detrusor muscle instability.

Objective To explore the effects and mechanism of gambogic acid ( GA) on lipopolysaccharide ( LPS)-induced acute lung injury ( ALI) .Methods ALI model was established by the injection of lipopolysaccharide into the tail vein of mice(4mg/kg, iv).The mice were randomly divided into control group (control), model group (model), GA group(GA), and pretreatment with GA of ALI group (GA+LPS).After six hours, the wet/dry weight ratio ( W/D) of the lung, myeloperoxidase ( MPO) activity in lung tissue , total proteins and number of white blood cells in bronchoalveolar lavage fluid ( BALF) were determined .Tumor necrosis factor-α( TNF-α) and interleukin-1β( IL-1β) were measured by the enzyme-linked immunosorbent assay methods .Results Compared with control group, the wet/dry weight ratio (W/D) of the lung, MPO activity in lung tissue, levels of total pro-teins and number of white blood cells in BALF of LPS group obviously increased , in addition the level of lung tissue TNF-αand IL-1βin LPS group were significantly higher (all P<0.01), while in GA pretreatment groups alleviated all the changes in ALI mice .Conclusions GA pre-treatment attenuated LPS-induced ALI , possibly by inhibiting inflammatory cytokines production so as to decrease the recruitment of neutrophils , reduce pulmonary edema .

Objective To design,synthesize and screen high efficient small interfering RNA(siRNA) targeting to cyclooxygenase-2(COX-2)on rheumatoid arthritis synovial fibroblasts(RASF).To further study the effect of specific COX-2 siRNA interfering on mediators of inflammatory cytokines.Methods Four pairs of siRNA for human COX-2 mRNA were synthesized by utilizing RNA design software,while another random sequence was designed as control.They were divided into group A to H.Among them,group A was used as the negative control(CTL),and group B to F were transfected as random siRNA(NC),1#～4#siRNA in order. These siRNAs were transferred into RASF by LipofectAMINE2000 package and PMA(phorbol-12-myristate- 13-acetate)was added into each culture and with a final concentration of 100 nmol/l.RASF was collected 48 hours after transfection.The expression of hCOX-2 at mRNA level was determined by reverse transcription- polymerase chain reaction(RT-PCR)and hCOX-2 protein level by Western Blot.The supernatant levels of PGE_2,IL-1?,IL-6,TNF-?and vascular endothelial growth factor(VEGF)of the above groups were detected by ELISA.Results The levels of hCOX mRNA and protein in RASF treated with 4-#siRNA were significantly lower than those of the negative control and other groups.The level of PGE_2 and cytokines like IL-1?,IL-6, TNF-?and VEGF in the supernatant were lower in the 4#siRNA group than in other groups.Conclusion 4#siRNA can effectively inhibit the expression of COX-2 mRNA and the synthesis of the COX-2 protein in human synovial fibroblasts.The level of PGE_2,IL-1?,IL-6,TNF-?and VEGF is the lowest in the super- natant.Thus 4#siRNA has been confirmed to specifically block the COX-2 in human synovial fibroblasts.

OBJECTIVETo validate transient elastography (Fibroscan) in assessment of hepatic fibrosis in autoimmune hepatitis (AIH).

METHODSLiver stiffness was assessed using Fibroscan in totally 30 patients with AIH. We compared the results of Fibroscan with the Scheuer fibrosis stage in liver biopsy in each patient.

RESULTS4 patients were shown as liver fibrosis stage S0, 6 as S1, 5 as S2, 11 as S3 and 4 as S4. Failure of the Fibroscan measurement occurred in 1 case (3.3%) because of her increased body mass index (BMI). The stiffness of Fibroscan was significantly correlated with the liver biopsy fibrosis stage (r = 0.801, P less than 0.001). The liver stiffnesses between mild and moderate fibrosis (S0-2) and advanced fibrosis (S3-4) were significantly different (t = -3.937, P = 0.001).

CONCLUSIONTransient elastography (Fibroscan) is a promising non-invasive method for detection of fibrosis in patients with autoimmune hepatitis. Its use for the follow up and management of these patients and should be evaluated further.

Elasticity Imaging Techniques ; methods ; Hepatitis, Autoimmune ; diagnostic imaging ; Humans ; Liver ; diagnostic imaging ; Liver Cirrhosis ; diagnostic imaging

OBJECTIVESTo evaluate the value of the metastatic to examined lymph nodes (rN) ratio in gastric cancer patients who underwent radical resection.

METHODSIn this retrospective study, data were collected from the medical records of 710 patients who underwent radical gastrectomy (R0) for gastric cancer from 1980 to 2006 in the Department of Surgical Oncology at the First Affiliated Hospital of China Medical University. The patients were divided into 2 groups according to the number of examined lymph nodes: Group 1 consisted of 327 patients with <15 examined lymph nodes and Group 2 consisted of 383 patients with ≥15 lymph nodes. rN categories staging and pN categories were divided separately according to the metastatic lymph node ratio and the examined lymph nodes. The prognostic factors were analyzed by univariate (Log-rank) and multivariate (Cox model) analysis methods.

RESULTSThe median survival time was 74 months (95% CI:55.6-92.4 months) in Group 1 and 96 months (95% CI:77.8-119.2 months) in Group 2, and the difference was not statistically significant (P>0.05). On multivariate analysis, the N ratio remained as an independent prognostic factor in both Group 1 (P<0.01, RR=1.225, 95% CI:1.102-1.362) and Group 2 (P<0.01, RR=1.421, 95% CI:1.269-1.592). However, pN stage was an independent prognostic factor only in Group 1. When the rN ratio classification was applied, there were no significant differences between each categories (P>0.05). However, the overall survival of patients with pN1 disease in Group 1 was significantly shorter than that in Group 2 according to the pN stage classification (P<0.01).

CONCLUSIONSThe metastatic lymph node ratio is an independent prognostic factor of the prognosis of gastric cancer. The staging system based on metastatic lymph node ratio (rN) is more reliable than the system based on the number of metastatic lymph nodes in the prediction of the prognosis of gastric cancer.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Stomach Neoplasms ; diagnosis ; pathology ; surgery