Animals ; Gallbladder ; physiology ; Rabbits ; Solitary Nucleus ; drug effects ; Thyrotropin-Releasing Hormone ; pharmacology ; Vagus Nerve ; drug effects ; physiology

· AIM: To search an easy and simple way for intraocular implantation after the eye evisceration.· METHODS: Fifteen healthy New Zealand rabbits were divided into 5 groups according to the sacrifice time, and each group included 3 rabbits; the left eye received the injection of polymethyl methacrylate (PMMA) bone cement (2g per mL), while the right eye served as control. Under general anesthetia, a 3mm incision was made on the sclera,and the eye contents and pigment tissues were extruded out with fingers. Then, PMMA bone cement (2g per ml) was injected through the scleral incision. Both the operated eye and control eye of the rabbits were enucleated and weighed,The reaction of the operated eye (macroscopically and histopathologically) was noted at frequent interval. The obtained data were then analyzed with ANOVA (SPSS11.5).· RESULTS: There was swelling of eyelids and conjunctiva at the early time after the injection, but no significant difference between the weight of the left and right eyes was noted,Histopathologic examination showed scleral and other tissues necrosis at early period, and then the tissues reaction turned into a great deal of cell proliferation and finally into extensive fibro-connective tissues. Three months after the operation,neovascularization was observed in the cornea of the operated eyes. Histopathologic examination showed formation of fibro-membrane around the intraocular implant,and disappearance of the inflammation.· CONCLUSION: The method of injecting PMMA bone cement (2g per ml) to form an intraocular implant is quite simple and economical; this method is also easy to use clinically.

Adult ; Electrocardiography ; Humans ; Male ; Myotonic Dystrophy ; complications ; pathology ; physiopathology ; Syringomyelia ; complications ; pathology ; physiopathology

BACKGROUNDCorneal neovascular leakage can lead to edema and secondary scarring. Previous studies have shown that pericytes play a key role in maturation of angiogenesis. The present studies investigate the relationship between vascular permeability and pericyte coverage of endothelial cells in rat corneal neovascular induced by alkali burns.

METHODSCorneal neovascular vessels induced by alkali burns was performed in Sprague-Dawley rats. Corneas were excised on 1, 2, 3, 5, 7 and 10 days after cauterization. The vascular permeability rate was measured by the Evans blue method. The microvessel pericyte coverage index (MPI) was applied to quantify the pericyte coverage through double immunofluorescent staining of frozen sections of corneas with CD31 as the endothelial and alpha-smooth muscle actin (alpha-SMA) as the pericyte markers. The correlation between permeability rate and MPI was analyzed. Pericyte coverage was confirmed ultrastructually using transmission electron microscopy.

RESULTSThe vascular permeability rate was (1.14 +/- 0.17), (0.24 +/- 0.08), (0.29 +/- 0.16), (0.14 +/- 0.10), (0.09 +/- 0.06) and (0.05 +/- 0.04) microg x ml(-1) x mm(-2) respectively on 1, 2, 3, 5, 7 and 10 days after cauterization. The MPI was 0, 16.07%, 11.95%, 43.84%, 73.97% and 86.21% respectively at the above mentioned time points. The correlation coefficient between MPI and the permeability rate was -0.943 (P = 0.005).

CONCLUSIONSPericyte recruitment was significantly correlated with the permeability of corneal neovascularization induced by alkali burns in rats. Therapeutic strategies aiming at anti-leakage should be most effective if they promote pericytes proliferation in the course of corneal neovascularization.

Alkalies ; Animals ; Burns, Chemical ; physiopathology ; Capillary Permeability ; Cell Movement ; Cornea ; blood supply ; ultrastructure ; Corneal Neovascularization ; physiopathology ; Eye Burns ; chemically induced ; physiopathology ; Female ; Fluorescent Antibody Technique ; Pericytes ; physiology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo construct a recombinant prokaryotic expression vector (plasmid) containing microneme protein 1 (MIC1) partial gene in toxoplasma gondii (T. gondii) ZS2 isolate. The gene was expressed in varied Escherichia coli (E. coli) after sequencing.

METHODSThe gene fragment coding MIC 1 from the genomic DNA of T. gondii ZS2 isolate was amplified by polymerase chain reaction (PCR). The gene was inserted to a prokaryotic expression vector pWR450-1 by digesting with restriction enzymes and linking reaction. The positive clone was screened on LB plates containing ampicillin and identified by restrictive enzyme digestion, PCR amplification and sequence analysis. The recombinant plasmid was transferred into E. coli TG1, JM109 (DE3) and DH5 alpha, and was expressed under the induction of IPTG. The expression products were identified by SDS-PAGE. The MIC1 gene structure was analyzed and compared in homology with the gene sequence of RH isolate using computer software.

RESULTSThe recombinant plasmid pWR450-1/MIC1, after cloning from acquired 471 bp MIC1 gene fragment and amplified from the genome gene ZS2, was complete homologous to the sequence of RH isolate, reflecting its highly conservative. The gene could be expressed as fusion protein with 70,000 in varied E. coli.

CONCLUSIONRecombinant plasmid pWR450-MIC1 was successfully constructed and could be expressed in different strains of E. coli, laying a foundation for research on its structure and function.

Animals ; Base Sequence ; Cell Adhesion Molecules ; biosynthesis ; genetics ; Cloning, Molecular ; DNA, Protozoan ; analysis ; Escherichia coli ; genetics ; Gene Expression ; Molecular Sequence Data ; Nucleic Acid Amplification Techniques ; Protozoan Proteins ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Toxoplasma ; genetics

Primary angiitis of the central nervous system is a rare and difficult entity. Here we represented the clinical and pathological features of a patient with little response to steroid before definite diagnosis. The 50-year-old male had a fluctuating disease course for more than 3 years. He presented visual disorders, seizure, cognitive impairment, hypersomnia, unsteady gait, dysphasia, dysphagia, and incontinence. Magnetic resonance imaging showed multiple, supratentorial and infratentorial abnormal signals, while cerebrospinal fluid and cerebral angiography were normal. Magnetic resonance spectrum showed a decrease of N-acetyl-aspartate. Brain biopsy revealed nongranulomatous lymphatic vasculitis with reactive gliosis, cicatrization, demyelination and focal hemorrhages.
Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Vasculitis, Central Nervous System ; diagnosis

OBJECTIVETo explore the effects of traditional Chinese herbal medicine Sijunzi decoction on amelioration of postburn intestinal injury in scalded rats.

METHODSOne hundred and eighty Wistar rats were randomly divided into 3 groups, i.e. scald and treatment (T), scald control (S) and normal control (C) groups. The rats in T group were gavaged with the decoction consisting of tangshen, tuckahoe, large head atractylodes rhizome, glycyrrhizic and rhubarb in a dose of 2 ml twice daily, while the rats in C group were just gavaged with the same amount of distilled water. The rats were sacrificed according to the scheduled postburn observation timepoints. The contents of TNF, NO, MDA and ATPase activity in rat plasma and the intestinal mucosa and the S-IgA content in the intestinal mucus were determined respectively. The changes in histopathology of intestinal mucosa were observed. The samples from internal organ tissue and blood were obtained for bacterial culture.

RESULTSThe contents of TNF, NO and MDA in the intestinal mucosa tissue and the rat plasma in scalded rats were lowered significantly by Sijunzi decoction. Furthermore, S-IgA secretion from intestinal mucous cells was maintained by Sijunzi decoction. T cell count was recovered and intestinal mucous barrier injury were lessened, and the bacterial positive rate in the internal organs was decreased.

CONCLUSIONTraditional Chinese herbal medicine Sijunzi decoction might be helpful in alleviation of postburn intestinal injury and in the prevention of intestinal bacterial translocation.

Animals ; Bacterial Translocation ; drug effects ; Burns ; blood ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Intestinal Mucosa ; metabolism ; pathology ; Intestines ; microbiology ; Male ; Rats ; Rats, Wistar

Objective To investigate the effects of the aqueous extracts of Eclipta Prostrata L.( EPL) on plasma lipid levels and the structure of vascular intima in rats with hyperhomocysteinemia ( Hhcy ) . Methods High L -methionine diet was used to construct HHcy rat model.Fifty SD rats were randomly divided into five groups:normal group, model group, lovastatin group (2 mg? kg -1 ), Eclipta Prostrata L.(EPL) low-and high-doses groups (5, 10 g? kg -1 ).The drags were administrated by intragastric administration once a day . After 8 weeks administration , the level of serum homocysteine ( Hcy ) , total cholesterol (TC) and trigly-ceride ( TG) were detected by biochemical analysis.While nitric oxide (NO), endothelin (ET) and thromboxane B2 ( TXB2 ) were measured by enzyme linked immunosorbent assay (ELISA).The histopathological examination of the thoracic aortas were observed microscopically afterhematoxylin -eosin ( HE ) staining. Results Compared with model group , serum levels of Hcy , TG, TC, ET and TXB 2 were significantly decreased ( P<0.05 ) in EPL groups and lovastatin group , while NO levels were significantly increased ( P<0.05) in high dose EPL group.HE sections showed that the shape of thoracic aortas looks as normal .Conclusion The aqueous extract of EPL could effectively reduce serum Hcy and regulate plasma lipid levels , thus it could alleviate vascular damage .

Monitoring engraftment of donor cells after allogeneic transplantation is the key of assessing successful establishment of animal transplantation model. The purpose of this study was to establish a method for analysis of chimerism in rhesus transplantation model. Y-specific sequence in rhesus was amplified by the polymerase chain reaction (PCR), method for analysis of chimerism in rhesus after sex-mismatched transplantation was established; the feasibility and sensitivity of the approach were tested by using serial DNA mixtures of sex-mismatched individuals; the accuracy of results was confirmed by chromosome karyotype analysis simultaneously; Chimerisms of one rhesus received allogeneic stem cell transplantation and the other received mesenchymal stem cells (MSC) transfusion were detected by this method. The results showed that a 176 bp long sequence of PCR product was gained in male rhesus, while no product was gained in female rhesus. The sensitivity of this method was up to 0.05% (male/female DNA ratio). Male donor chimerism were found on day 7 and 14 after allogeneic stem cell transplantation by Y-specific sequence and chromosome karyotype analysis. Otherwise, male donor chimerism was found in peripheral blood at 1 hour and in bone marrow on day 30 after MSC transfusion by this method, but no male donor chimerism was found after MSC transfusion using chromosome karyotype analysis. In conclusion, this rapid, sensitive approach can used to assess chimerism in experiments of rhesus alloorgan transplantation and cell transfusion.
Animals ; Base Sequence ; Female ; Macaca mulatta ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Models, Animal ; Molecular Sequence Data ; Transplantation Chimera ; blood ; genetics ; Transplantation, Homologous ; Y Chromosome ; genetics

OBJECTIVETo study the role of mitogen activated protein kinase (MAPK)/activator protein-1 (AP-1) pathway in benzo(a)pyrene (B(a)P)-induced changes of cell cycle in human embryo lung fibroblasts (HELF).

METHODSAP-1 luciferase activity was determined by the Luciferase reporter gene assay using a luminometer. The expression levels and activity of extracellular signal-regulated protein kinase (ERK), c-Jun NH2-terminal kinase (JNK) and p38 were determined by Western blot. Flow cytometric analysis was employed to detect the distributions of cell cycle. The dominant negative mutant of ERK2, JNK1 and p38 were applied to detect the upstream or downstream relationship of signaling pathways.

RESULTSB(a)P treatment resulted in a marked activation of AP-1 and its upstream MAPK, including ERK, JNK and p38 in human embryo lung fibroblasts (HELF). B(a)P exposure also led to increase the population of cells at S phase compared to control (P < 0.01) with a concomitant decline of cells at G(1) phase. B(a)P-induced cell cycle alternation was markedly impaired by stable expression of a dominant negative mutant of ERK2 or JNK1, but not p38. B(a)P-induced AP-1 transactivation was inhibited by the overexpression of dominant-negative mutant of ERK2 or JNK1, but not p38. Inhibition of the activation of AP-1 by curcumin, a chemical inhibitor of AP-1, significantly inhibited the cell cycle changes in response to B(a)P treatment.

CONCLUSIONERK and JNK, but not p38, mediated benzo(a)pyrene-induced cell cycle changes by AP-1 transactivation in HELF.

Benzo(a)pyrene ; pharmacology ; Blotting, Western ; Cell Cycle ; drug effects ; Cells, Cultured ; Fibroblasts ; cytology ; drug effects ; metabolism ; Flow Cytometry ; Humans ; Lung ; cytology ; embryology ; Mitogen-Activated Protein Kinase 1 ; metabolism ; physiology ; Mitogen-Activated Protein Kinase 8 ; metabolism ; physiology ; Phosphorylation ; Transcription Factor AP-1 ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism