Objective To investigate the effects of mild hypothermia ( MH ) on the expression of my- eloperoxidase(MPO) and cyclooxygenase 2 (COX 2) in rats after cerebral ischemia and reperfusion (CIR). Methods Forty-eight Wistar rats were randomly divided into four control groups (n=6 in each) and four MH groups (n=6 in each).CIR models were established by suture occlusion of the left middle cerebral artery.The rats in the MH groups,but not in the control groups,were treated with MH.Rats were killed at 4 h,8 h,12 h and 16 h after CIR.MPO expression was measured,along with the expression of COX 2 as measured by Western blot- ting and immunohistochemical methods.Results Compared with the control groups,MPO activity and the COX 2 expression in the cortex and striatum were significantly lower in all the MH groups at 4 h,8 h,12 h and 16 h after CIR.Conclusion MH treatment can protect neurons by decreasing MPO activity and COX 2 expression,allevia- ting inflammation and reducing secondary injuries after CIR.

OBJECTIVETo isolate and characterize the dental follicle cells (DFC) from dental follicle (DF) tissues of normal human impacted third molars.

METHODSDFC were isolated from the DF tissues of healthy young human impacted third molars. A limited dilution culture was used to assess DFC colony-forming efficiency. The expressions of Stro-1, Notch-1 and nestin in DFC were detected by immunohistochemistry analysis. The primary DFC cultures were subjected to a variety of treatment modes: osteogenic, adipogenic and chondrogenic differentiation. DFC and periodontal ligament cells (PDLC) proliferation abilities were compared by methyl thiazolyl tetrazolium (MTT) assay. The expressions of tenascin-N and F-spondin in DFC and PDLC were evaluated by reverse transcriptase polymerase chain reaction (RT-PCR).

RESULTSMost DFC were spindle fibroblast-like cells. DFC cultures formed colonies from passage 1 cells and the frequency of colony forming efficiency (CFE) was 3.70%. Some of the DFC were stained positively for Stro-1 and almost all the DFC were stained positively for Notch-1 and nestin. DFC cultures displayed multipotential characteristics following fate-specific inductions for 21 days. Alizarin red positive condensed nodules were detected following osteogenic induction, oil red-positive lipid vacuoles were generated using adipogenic induction and collagen-II was revealed following chondrogenic induction by immunohistochemistry. On day 3 and 5, DFC (0.20 ± 0.01, 0.51 ± 0.09) showed a better cell activity than PDLC (0.16 ± 0.03, 0.47 ± 0.07) (P > 0.05). On day 7, DFC (1.03 ± 0.11) exhibited a higher proliferation rate than PDLC (0.93 ± 0.09) (P < 0.05). RT-PCR results showed that tenascin-N was not expressed in DFC, but expressed moderately in PDLC. F-spondin was expressed strongly in DFC, while not expressed in PDLC.

CONCLUSIONSDFC from ectomesenchymal tissues showed a good viability and contained cells similar to the mesenchymal stem cells. It may be used as a novel cell source for periodontium regeneration.

Adolescent ; Antigens, Surface ; metabolism ; Cell Differentiation ; Cell Separation ; Cells, Cultured ; Dental Sac ; cytology ; Extracellular Matrix Proteins ; metabolism ; Female ; Humans ; Male ; Molar ; Nestin ; metabolism ; Receptor, Notch1 ; metabolism ; Stem Cells ; cytology ; Tenascin ; metabolism ; Tissue Engineering ; Tooth, Impacted ; Young Adult

Animals ; Calcium Channel Blockers ; therapeutic use ; Dogs ; Hypothermia ; complications ; Male ; Nimodipine ; therapeutic use ; Ventricular Fibrillation ; etiology ; prevention & control

OBJECTIVE: To investigate the numbers, sizes and types distribution of diatoms in drowned and postmortem immersed rabbits' lungs. METHODS: Sixty-two rabbits were randomly divided into drowning group (n = 30), postmortem immersion group (n = 30) and land death group (n=2), and the diatoms in each lung lobe were analyzed quantitatively and qualitatively by microwave digestion and scanning electron microscopy. RESULTS: In the drowning group, the diatoms were detected in each lung lobe with Cyclotella and Melosira in the majority. In the postmortem immersion group, Cyclotella was in the majority. And the diatoms weren't detected in some lung lobes in postmortem immersion. There were significant differences in the detection rates of upper lobe of left lung, middle lobe and cardiac lobe of right lung in two groups (P < 0.05). CONCLUSION Based on the microwave digestion and scanning electron microscopy, the numbers, sizes and types distribution of diatoms in drowned and postmortem immersed rabbits' lungs can be analyzed and used as references for testing theory.
Animals ; Autopsy ; Diatoms/isolation & purification* ; Drowning ; Lung/microbiology* ; Microscopy, Electron, Scanning ; Microwaves ; Rabbits

AIMTo synthesize new fluoroquinolone analogues as antibacterial compounds.

METHODS AND RESULTSBy reaction of acryl chloride(chloro-carbonic ester) with sodium sulfocyanate, acyl isosulfocyanic ester were easily obtained. Twelve 7-(4-acylamino-thiocarbamoyl-1-piperazinyl) fluoroquinolone analogues (1-12) were synthesized through modifying the 7-piperazine of norflorxacin and ciprofloxacin with isosulfocyanic ester synthesized above. The structures of synthesized compounds were characterized by 1HNMR, IR and elemental analysis.

CONCLUSIONAntibacterial activities of the new compounds were evaluated in vitro compared with norflorxacin. Compounds 5, 7, 10 and 12 showed antibacterial activities.

Anti-Infective Agents ; chemical synthesis ; chemistry ; pharmacology ; Bacillus subtilis ; drug effects ; Ciprofloxacin ; chemistry ; pharmacology ; Combinatorial Chemistry Techniques ; methods ; Escherichia coli ; drug effects ; Fluoroquinolones ; chemical synthesis ; chemistry ; pharmacology ; Microbial Sensitivity Tests ; Molecular Structure ; Norfloxacin ; chemistry ; pharmacology

OBJECTIVETo find the function and functioning mechanism of Yiqi Qingwen Jiedu Heji in resisting influenza immune damage by studying its effect on the expressions of cytokine.

METHODTaking IV FM1 infected mice as its model and doing ELISA (double antibody sandwichenzyme linked immunosorbent assay), we dynamically observed the change of cytokine TNF-alpha, IL-6, IFN-gamma and IL-10 after giving Yiqi Qingwen Jiedu Heji treatment.

RESULT AND CONCLUSIONAfter the mice are infected by influenza virus, their protein expressions of the model group are higher than those of the control group, of which TNF-alpha, IL-6, IFN-gamma reach the peak in three days. The three expressions of Yiqi Qingwen Jiedu Heji treated group are decreased and the decrease becomes remarkable on the third day, compared with those of the model group. However, the expression of IL-10 of the treated group is remarkably increased. It indicates that Yiqi Qingwen Jiedu Heji can resist the expressions of TNF-alpha, IL-6 and IFN-gamma pro-inflammatory cytokine,increase the expression of IL-10, and thus, alleviate inflammatory injury. So the clinical application of such medicine can shorten the course of disease.

Animals ; Cytokines ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-6 ; metabolism ; Lung ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Orthomyxoviridae ; Orthomyxoviridae Infections ; metabolism ; virology ; Plants, Medicinal ; chemistry ; Tumor Necrosis Factor-alpha ; metabolism

Aged ; Echinocandins ; administration & dosage ; therapeutic use ; Humans ; Leukemia, Myelomonocytic, Chronic ; pathology ; Lipopeptides ; Male ; Pneumonia, Pneumocystis ; diagnosis ; drug therapy ; pathology ; Trimethoprim, Sulfamethoxazole Drug Combination ; administration & dosage ; therapeutic use ; Uremia ; pathology

Many antineoplastic agents can cause myelosuppression and thrombocytopenia. Thrombopoietin (TPO) is believed to be the major cytokine affecting the proliferation and maturation of megakaryocytes and increasing circulating platelet levels. We have designed and synthesized a TPO mimetic peptide, it can increase circulating platelet levels in vivo. For increasing half-life and forming dimer, the peptide was expressed as chimeric proteins with human IgG1 Fc fragments. The cDNA of TPO mimetic peptide was synthesized chemically and linked respectively to 5' terminus of human IgG1 Fc cDNA fragments in various length (Fc1: Fc 5' 648 bp; Fc2: Fc 5' 270 bp; Fc3: Fc 5' 267 bp; Fc4: Fc 5' 90 bp), and cloned into expression plasmid pET28a (+) for constructing four recombinant plasmids. By transforming the four recombinant plasmids into E. coli. BL21 (DE3) respectively, we got 3 kinds of engineered E. coli which express TPO + Fc chimeric proteins(28 kD TPO + Fc1, 12 kD TPO + Fc2 and 12 kD TPO + Fc3) at high level respectively, the expressed proteins were purified with DEAE-Sepharose FF and S-Sepharose FF column. The bioactivities of the expressed chimeric proteins(TPO + Fc1, TPO + Fc2 and TPO + Fc3), TPO mimetic peptide, and PEG4000 coupled TPO mimetic peptide were evaluated with Ba/F3-mp1 in vitro and with carboplatin-induced thrombocytopenia mice in vivo, the expressed chimeric proteins have higher activity than TPO mimetic peptide both in vitro and in vivo, the EC50 on Ba/F3-mp1 cells were 13, 10, 10, 50, and 25 nmol/L respectively, all of them can increase circulating platelet counts. Their imol/Lunogenicity were valuated in mice, none of them can elicit mice to produce antibodies to TPO mimetic peptide, meanwhile three TPO + Fc chimeric proteins can elicit mice to produce antibodies to human IgG1 Fc. These studies have laid basis for production of TPO mimetic peptide by genetic engineering.
Animals ; Base Sequence ; Blood Platelets ; drug effects ; Cell Division ; drug effects ; Female ; Humans ; Immunoglobulin Fc Fragments ; genetics ; immunology ; pharmacology ; Immunoglobulin G ; genetics ; Male ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Oligopeptides ; chemical synthesis ; genetics ; pharmacology ; Recombinant Fusion Proteins ; genetics ; immunology ; pharmacology ; Thrombocytopenia ; blood ; immunology ; Thrombopoietin ; genetics ; immunology ; pharmacology

The DNA coding for the fusion protein of thromobopoietin mimetic peptide (TMP) and human IgG1 Fc fragment was amplified from recombinant plasmid pET28a/TMPFc, inserted into pPICZalphaA and transformed into Pichia pastoris using electroporation. The recombinants of correct phenotype were identified after screening on MDH and MMH culture medium. The fusion gene was verified with PCR and western blot. MTT method was used to test the activity of TMPFc in promoting the growth of Ba/ F3-mpl cell. The TMPFc with a 64 000 molecular weight was a secretary protein in the system and its expression amounted to 65% of the total protein in the medium supernatant. The TMPFc showed a promotive effect on the growth of Ba/F3-mpl in vitro. A significant portion of the secretary protein existed as dimer, which provided material for studying the dimer in future.
Amino Acid Sequence ; Blotting, Western ; Humans ; Immunoglobulin Fc Fragments ; genetics ; Immunoglobulin G ; genetics ; Molecular Sequence Data ; Pichia ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; biosynthesis ; Thrombopoietin ; genetics

Based on thermodynamic principle, the critical relative humidity of electrolytes is closely related to their solubility. The authors explored the relationship theoretically and calculated critical relative humidity of 21 electrolytes from their solubility in the light of Raoult's law and extended Wilson model. The results indicate that the critical relative humidity values calculated by Raoult's law can not accord with the reported ones and there is a systematic error in the high concentration range; while these calculated by extended Wilson model are comparable to the reported ones.
Electrolytes ; chemistry ; Humidity ; Models, Chemical ; Solubility