OBJECTIVETo observe the role and mechanism of CO-releasing molecule (CORM)-2 in lung injury induced by ischemia-reperfusion (IR) of hind limbs in rats.

METHODSA rat model of lung injury induced by IR of hind limbs was established. A total of 40 Sprague Dawley (SD) rats were randomly divided into 5 groups (n equal to 8): sham, sham + CORM-2, IR, IR + CORM-2 and IR + dimethyl sulfoxide (DMSO). Rats in the IR group received hind limb ischemia for 2 hours and reperfusion for 2 hours, rats in the sham group underwent sham surgery without infrarenal aorta occlusion, rats in the IR+CORM-2 group and in the sham + CORM-2 group were given CORM-2 (10 micromol/kg intravenous bolus) 5 minutes before reperfusion or at the corresponding time points, while rats in the IR + DMSO group was treated with the same dose of vehicle (DMSO) at the same time. The lung tissue structure, polymorphonuclear neutrophil (PMN) count, wet-to-dry weight ratio (W/D), malondialdehyde (MDA) content, myeloperoxidase (MPO) activity, intercellular adhesion molecule-1 (ICAM-1) expression,IkBa degradation and nuclear factor (NF)-kB activity in the lungs were assessed.

RESULTSAs compared with the sham group, lung PMNs number, W/D, MDA content, MPO activity, ICAM-1 expression and NF-kB activity significantly increased in the IR group, but the level of IkBa decresed (P less than 0.01). Compared with the IR group, lung PMNs number, W/D, MDA content, MPO activity and ICAM-1 expression significantly decreased in the IR+COMR-2 group (P less than 0.01), while the level of IkBa increased.

CONCLUSIONSThese data demonstrate that CORM-2 attenuates limb IR-induced lung injury through inhibiting ICAM-1 protein expression, NF-kB pathway and the leukocytes sequestration in the lungs following limb IR in rats, suggesting that CORM-2 may be used as a therapeutic agent against lung injury induced by limb IR.

Animals ; Hindlimb ; blood supply ; Immunohistochemistry ; Intercellular Adhesion Molecule-1 ; metabolism ; Lung Injury ; etiology ; metabolism ; Male ; NF-kappa B ; metabolism ; Neutrophils ; metabolism ; Organometallic Compounds ; pharmacology ; Rats ; Reperfusion Injury ; complications

OBJECTIVETo investigate the anti-inflammatory effects on the vessel wall of rosuvastatin in apolipoprotein E-deficient mice.

METHODSEight-week-old apolipoprotein E-deficient mice fed a normal chow diet were treated with vehicle or various doses of rosuvastatin (1, 5, or 20 mg/kg) by subcutaneous injection for 2 or 6 weeks prior to sacrifice. Endothelial adhesiveness for monocytes was determined by functional binding assay. The expressions of vascular cell adhesion molecule-1 and monocyte chemotactic protein-1 in the vessel wall were detected by quantitative real-time polymerase chain reaction.

RESULTSEndothelial adhesiveness for monocytes was significantly attenuated after 2 or 6 weeks treatments with 5 or 20 mg/kg rosuvastatin. Rosuvastatin also significantly reduced the expressions of vascular cell adhesion molecule-1 and monocyte chemotactic protein-1 in the vessel wall.

CONCLUSIONThe anti-inflammatory effects of suvastatin might be responsible for attenuating the pathogenesis of atherogenesis in apolipoprotein E-deficient mice.

Animals ; Apolipoproteins E ; genetics ; Cell Adhesion ; drug effects ; Endothelium, Vascular ; cytology ; Fluorobenzenes ; pharmacology ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Monocytes ; drug effects ; metabolism ; Pyrimidines ; pharmacology ; Rosuvastatin Calcium ; Sulfonamides ; pharmacology ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo investigate SiO2-induced EMT in human bronchial epithelial cells HBE in vitro.

METHODSHBE cells were cultured and then stimulated with indicated doses of SiO2 (0, 50, 100, 200, 300 µg/ml). The morphological changes were observed by microscope. In addition, Western blot was per-formed to detect the expression of E-cad, α-SMA and Vim. The changes of migration ability were examined by wound-healing assay in vitro.

RESULTS(1) After exposure to SiO2, HBE cells lost contact with their neighbor and displayed a spindle-shape, fibroblast-like morphology. (2) Compared with the control, the E-cad (300 µg/ml group) expression downregulated 2.98 fold (P < 0.05), and the Vim (300 µg/ml group) and α-SMA (200 µg/ml group) expression upregulated 4.46 fold and 3.55 fold (P < 0.05). There were significant differences between 100, 200, 300 µg/ml groups and the control group (P < 0.05). (3) In the test group, the percentage of wound-healing areas/wound areas were larger than those in control group (P < 0.05).

CONCLUSIONSSiO2 could induce EMT in human bronchial epithelial cells.

Bronchi ; cytology ; Cells, Cultured ; Epithelial Cells ; cytology ; drug effects ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Silicon Dioxide ; adverse effects ; Stromal Cells ; cytology ; drug effects

OBJECTIVETo investigate the diagnostic methods and reasonable treatment of Peutz-Jeghers syndrome (PJS).

METHODSClinical data of six patients with PJS were reviewed.

RESULTSRepeated abdominal pain, intussusception and intestinal polyp with bleeding were main manifestations. Four patients father,three patients grandfather and one patients mother were diagnosed with PJS. Three patients had family history of cancer. Case 4 and case 5 underwent laparotomy for many times because of intussusceptions caused by polyps or recurrent abdominal pain. Case 1 and case 4 had polyps synchronous with adenoma, and case 2 had polyp with gastric cancer. Main treatment included polyp resection and partial small intestinal and colon resection.

CONCLUSIONSPatients with PJS have family history of cancer and a high incidence of polyp recurrence of small intestine. Surgical intervention is the first choice regimen. Surveillance should be emphasized on gastrointestinal tract and other potential malignant organs in PJS patients.

Adolescent ; Adult ; Female ; Humans ; Intestine, Small ; surgery ; Male ; Pedigree ; Peutz-Jeghers Syndrome ; diagnosis ; genetics ; surgery

OBJECTIVETo study the role of cerebrovascular hemodynamic indexes (CVHI) changing in stroke and to provide reference for stroke prevention and risk factor study.

METHODSFrom 2003 to 2004, participants aged 40 years and above in two communities in Fengxian district were recruited by cluster sampling. Risk factors of stroke and CVHI were investigated and checked during baseline investigation. A total of 10 565 individuals completed the survey and met the inclusion criterion. After baseline investigation, the cohort was followed up for stroke occurrence. Relative risk (RR) of CVHI and common risk factors were estimated by cohort study design.

RESULTSAge of the cohort was (56.2 ± 11.4) years. 4444 (42.1%) were males and 6121 (57.9%) were females. Total follow-up duration was 67 885.7 person-years. A total of 195 stroke cases occurred and incidence density of stroke was 287.2 per 100 000 person-years. Stroke incidence in exposure groups of hypertension, heart disease and alcohol drinking was 3.47% (108/3118), 2.96% (21/710) and 2.50% (47/1882), respectively. The incidence in corresponding non-exposure group was 1.17% (87/7448), 1.77% (174/9855) and 1.70% (148/8683) respectively. There was significant difference between 2 groups (χ(2) value was 62.72, 4.56 and 4.94, respectively, P < 0.05). Stroke incidence in CVHI score < 25, 25 - 49, 50 - 74 and ≥ 75 groups was 9.12% (59/647), 5.68% (44/775), 2.52% (39/1545) and 0.72% (53/7403)(χ(2)trend = 273.57, P < 0.05), respectively. Incidence of stroke in 40 - 49, 50 - 59, 60 - 69, ≥ 70 years age group was 0.22% (8/3565), 1.28% (43/3357), 2.71% (50/1848) and 5.88% (94/1600) (χ(2)trend = 181.48, P < 0.05), respectively. Multiple Cox regression analysis indicated that RR (95%CI) value of hypertension and cigarette smoking was 1.40(1.02 - 1.92) and 1.59(1.19 - 2.12), respectively when comparing with non-exposure group. RR (95%CI) value in CVHI score < 25, 25 - 49 and 50 - 74 points group were 6.15 (4.08 - 9.26), 4.55 (2.98 - 6.96) and 2.68 (1.75 - 4.09), respectively when comparing with the score ≥ 75 points group. RR (95%CI) value in age 50 - 59, 60 - 69 and ≥ 70 years group was 4.61 (2.16 - 9.82), 7.81 (3.67 - 16.60) and 13.49(6.44 - 28.24), respectively when comparing with below 40 years group.

CONCLUSIONCVHI score is the strong independent predictive factor and hypertension, cigarette smoking and age are the independent risk factors of stroke.

Aged ; Brain ; physiopathology ; Cohort Studies ; Female ; Hemodynamics ; Humans ; Male ; Middle Aged ; Risk Factors ; Stroke ; epidemiology ; etiology ; physiopathology

In order to confirm the reasonability of designed recombinant exotoxin B7-1-Linker-PE40 and B7-2-Linker-PE40, their molecular biology characteristics, such as flexibility, antigenicity, hydrophilicity, epitope and secondary structure, were predicted by using a computer software GOLDKEY. It had been found that the recombinant fusion exotoxin had kept the epitope characterstics of B7-1, B7-2 and PE40, and had not got new epitope, and the antigenicity in flexible linker was extxemely low. The linker inserted in the recombinant fusion exotoxin had low epitope, low antigenicity and high flexibility. Compared to B7-1, B7-2 and PE40, there are several amino acid residues changes in B7-1-Linker-PE40 and B7-2-Linker-PE40, respectively, which might have some effect on secondary structure of the recombinant fusion exotoxins. Western blot analysis revealed that both B7-1-Linker-PE40 and B7-2-Linker-PE40 could bind specifically with antibodies against B7-1, B7-2 and PE40, respectively. The result of Western blot was consistant with the computer prediction that the recombinant proteins retain the antigenicity and spacial structure of B7 and PE40. It is suggested that both fusion proteins designed and constructed were resonable and computer analysis would be helpful for us to study the biological activity of the recombinant fusion exotoxin B7-1-Linker-PE40 and B7-2-Linker-PE40 and construct other recombinant proteins further.

BACKGROUNDAutoimmune encephalitis associated with antibodies against γ-aminobutyric acid B receptor (GABA B R) in patients with limbic encephalitis (LE) was first described in 2010. We present a series of Han Chinese patients for further clinical refinement.

METHODSSerum and cerebrospinal fluid (CSF) samples from patients referred to the program of encephalitis and paraneoplastic syndrome of Peking Union Medical College Hospital were tested with indirect immunofluorescence. Clinical information of patients with anti-GABA B R antibody positivity was retrospectively reviewed, and descriptive statistical analysis was performed.

RESULTSAll eighteen anti-GABA B R antibody-positive cases had limbic syndromes, and electroencephalogram (EEG) or neuroimaging evidence fulfilled the diagnostic criteria of LE. Four patients had additional antibodies against Hu in serum and one had anti-N-methyl-d-aspartate receptor antibody in both sera and CSF. Seventeen (17/18) patients presented with new-onset refractory seizure or status epileptics. Twelve (12/18) patients had memory deficits, 11 (11/18) patients had personality change, 7 (7/18) patients had disturbance of consciousness, and 3 (3/18) patients showed cerebellar dysfunction. One patient with LE had progressive motor and sensory polyneuropathy. Lung cancer was detected in 6 (6/18) patients. Ten (10/18) patients showed abnormality in bilateral or unilateral mediotemporal region on magnetic resonance imaging. Ten (10/18) patients had temporal lobe epileptic activity with or without general slowing on EEG. Seventeen patients received immunotherapy and 15 of them showed neurological improvement. Four patients with lung cancer died within 1-12 months due to neoplastic complications.

CONCLUSIONSOur study demonstrates that most Han Chinese patients with anti-GABA B R antibody-associated LE have prominent refractory epilepsy and show neurological improvement on immunotherapy. Patients with underlying lung tumor have a relatively poor prognosis. Testing for anti-GABA B R antibodies is necessary for patients with possible LE or new-onset epilepsy with unknown etiology.

Adult ; Autoantibodies ; immunology ; China ; Electroencephalography ; Epilepsy ; immunology ; pathology ; Female ; Humans ; Limbic Encephalitis ; immunology ; pathology ; Male ; Middle Aged ; Receptors, N-Methyl-D-Aspartate ; immunology ; Retrospective Studies ; gamma-Aminobutyric Acid ; metabolism

OBJECTIVETo investigate device implanted complications and corresponding therapeutic strategies of Coflex interspinous dynamic stabilization system for lumbar spine intraoperatively and postoperatively.

METHODSFrom September 2008 to August 2010, 133 cases of degenerative disease of lumbar spine including 62 males and 71 females, ranging from 35 to 81 years of age (mean 60.8 years), underwent or planed to be underwent decompression with Coflex interspinous dynamic stabilization system were reviewed retrospectively, and 13 cases including 6 males and 7 females, ranging from 41 to 71 years of age (mean 58.6 years), occurred device implanted complications. The Coflex implanted complications were analyzed, and therapeutic strategies according to different character were carried out, scores of visual analogue scale (VAS), Oswestry disability index(ODI) and effect-related data preoperatively, postoperatively, after conservative treatment and in final follow-up were evaluated with paired-samples t test.

RESULTSThirteen cases of Coflex implanted complications and treatment applied included: 3 cases occurred fracture of spinous processes intraoperatively were treated by pedicle screws instead; 2 cases occurred fracture of spinous processes postoperatively or during follow-up, including 1 case underwent revision with pedicle screws, another 1 case treated with conservative treatment; 4 cases with degenerative coronal spondylolysis in surgical segments, 1 case with sagittal instability preoperatively, and 1 case with device dislodgment in follow-up all suffered aggravated pain and received conservative treatment; 1 case suffered implanted malposition intraoperatively was underwent internal fixation with pedicle screws instead; at length, 1 case with aggravated pain postoperatively and without definite reason received revision with internal fixation of pedicle screws demolishing the Coflex. The follow-up time of 13 cases ranged from 20 to 38 months (mean 27.6 months); and 7 cases implanted Coflex with aggravated pain of lumbar and lower limb, but the position of device can still maintained, were received conservative treatment, and whose score of VAS and ODI in the final follow-up were 1.9 ± 0.7 and 23.2 ± 3.4, and comparing to 6.1 ± 1.1 and 58.1 ± 3.0 preoperatively, evident improvement was got finally (t = 8.2 and 18.2, P < 0.01). Scores of VAS and ODI of 2 cases with Coflex implanted complications underwent revision with pedicle screws were also improved correspondingly.

CONCLUSIONSCoflex interspinous dynamic stabilization system implanted should be avoided to cases who suffered with osteoporosis, too narrow interspinous space and intervertebral coronal spondylolysis or sagittal instability; and choice of device, depth of implantation and intensity of clumping should be appropriate. For patients with symptom but device still in right position, conservative treatment can be carried out; but for patients subjected to malposition of device, failure of implantation intraoperatively or intolerance to device, revisions and salvages should be underwent with internal fixation of pedicle screws.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Internal Fixators ; adverse effects ; Intervertebral Disc Degeneration ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Postoperative Complications ; Retrospective Studies ; Spinal Fusion ; adverse effects ; instrumentation ; methods ; Treatment Outcome

OBJECTIVETo investigate the roles of Rho/Rock signaling pathway in silica-induced Epithelial-mesenchymal transition (EMT) in human bronchial epithelial cells (BEC) in vitro.

METHODSHuman BEC were incubated with silica with various concentrations for indicated times. Cell viability was assayed by MTT test. Morphologic Changes were observed by microscope. Mesenchymal marker α-smooth muscle actin (α-SMA), vimentin (Vim), and epithelial marker E-cadherin (E-cad) were analyzed by Western Blot. The pull-down assay was used to measure Rho activity. In the prevention experiments, the specific inhibitor for Rho effector ROCK (Y27632) was used to inhibit the activity of Rho.

RESULTSHuman BEC stimulated with silica were converted from a "cobblestone" epithelial structure into an elongated fibroblast-like shape structure. Incubation of human BEC with silica induced de novo expression of α-SMA and Vim, and loss of E-cad. Also, silica treatment resulted in Rho activation in human BEC. Y27632 up-regulated the E-cad expression but attenuated α-SMA and Vim expression in silica-stimulated cells.

CONCLUSIONThe activation of Rho/ROCK signaling pathways is most likely involved in Silica-induced EMT in human bronchial epithelial cells.

Actins ; metabolism ; Bronchi ; cytology ; Cadherins ; metabolism ; Cells, Cultured ; Epithelial Cells ; drug effects ; metabolism ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Quartz ; toxicity ; Signal Transduction ; Vimentin ; metabolism ; rho-Associated Kinases ; metabolism

OBJECTIVETo construct an expression vector of anti-MM scFv-tP fusion protein and test its expression efficiency and function.

METHODSThe truncated protamine (tP) gene sequence was added to the gene of single chain antibody against the specific antigen on the surface of malignant melanoma tumor cells using PCR. A GST-fusion expression vector was constructed and the soluable protein was expressed in the E.coli system. After cleavage and purification, the purified fusion protein was obtained. The binding activity of Anti-MM scFv-tP and siRNA was detected by EMSA. Flow cytometry and confocal microscopy were used to detect the cell surface antigen binding activity of the fusion protein.

RESULTSThe expression vector of Anti-MM scFv-tP fusion protein was successfully constructed. The soluable protein could be expressed in the E.coli system, and the purified fusion protein was obtained. The anti-MM scFv-tP fusion protein retained siRNA binding ability and could directly target malignant melanoma (MM) LiBr cells.

CONCLUSIONThe recombinant GST- Anti-MM-scFv-tp expression vector was successfully constructed. The fusion protein retains siRNA binding ability and can directly target LiBr cells to provide a reliable tool for further study.