Female ; Humans ; Infant ; Mastocytosis, Systemic ; diagnosis ; pathology ; therapy

Objective To examine the localization of neuronal specific transcription factor DAT1 mRNA in the central nervous system of the rat. Methods In situ hybridization histochemistry staining method with digoxigen-labeled cRNA probe was used. Results The Transcripts of DAT1 mRNA were localized in somatic and dendritic profiles at most regions of adult rat central nervous system. It can be observed in cerebrum, cerebellum, thalamus, brain stem and spine. The intense hybridization signal can be seen in olfactory bulb, entorhinal cortex, prepirform cortex, striate cortex, hippocampus CA1 and dentate gyrus, lateral dorsal nucleus of thalamus, red nucleus, dorsal tegmental nucleus, central reticular nucleus of medulla oblongata,motor nucleus of trigemina, nucleus of hypoglossal, vagus nerve nucleus, external cuneate nucleus and ambiguous nucleus. The moderate staining was detected in Ⅱ-Ⅵ layer of cerebrum, hippocampus CA2 and CA3, amygdala nucleus, globus pallidus, medial geniculate body, lateral geniculate body, zona incerta, supraoptic nucleus,paraventricular nucleus, arcuate nucleus, substantia nigra, mesencephalon reticular nucleus, reticular nuclei of pons, gigantocellular reticular nucleus, lateral reticular nucleus, medial nucleus of the trapezoid body, vestibular nucleus, dorsal cochlear nucleus, locus ceruleus, cuneate nucleus, nucleus raphes dorsalis. The faint signal showed in septal nuclei, ventral lateral nucleus of thalamus, ventral posterior nucleus of thalamus, posterolateral nucleus of thalamus, dorsomedial nucleus of thalamus, anterodorsal nucleus of thalamus, premammillary nucleus, central gray, superficial gray layer of the superior colliculeus, central nucleus of the inferior colliculeus, lateral nucleus of the inferior colliculeus, trigeminal mesencephalic nucleus, spinal nucleus of trigeminal, nucleus of solitary tract, inferior olivary nucleus, superior central nucleus, cerebellar fastigial nucleus, interposed cerebellared nucleus, lateral cerebellellar nucleus and spinal cord gray matter. Conclusion The results demonstrated the widely presence of DAT1 mRNA in adult rat central nervous system ,close related to the dopaminergic nervous system, suggesting a role of regulation for this gene in various functions of rat central nervous system, especially in dopamine neurotransmitter regulation.

Muscle recently has been identified as a good source of adult stem cells that can differentiate into cells of different lineages.Researchers have identified two types of stem cells in skeletal muscle.Further research is necessary to delineate the relationship between different populations of musclederived stem cells(MDSCs)and between MDSCs and other adult stem cells.The methods used to isolate these cells appear to influence the stem cell characteristics.As these efforts continue,the potential for MDSCsbased therapy for other musculoskeletal injuries,as well as for cardiac and smooth muscle injuries,is currently being explored.The behavior,biocharacteristic,isolation,differentiation and the probability of application to regenerate lost or diseased tissue of MDSCs were summarized.

Objective To explore the rare primary mt-DNA mutation in LHON patients in China.Design Clinical study.Partici- pants Clinically diagnosed LHON patients whose common primary mt-DNA mutations were negative and normal persons.Methods Polymerase chain reaction(PCR)and DNA sequencing were used to detect the mutations of G11778A,G3460A and T14484C in mt-DNA of clinically diagnosed LHON patients.Fragments of mt-DNA 3962～4356 and 11320～11789 were PCR amplified and se- quenced if all the three mutations above were negative and also in the control group.The sequence results were analyzed by the BLAST service provided by NCBI and compared with the 2001 Revised Cambridge Reference Sequence for the mt-DNA mutations C4171A and G11696A.Main Outcome Measures The sequence results of mt-DNA.Results There were totally 56 eases of clinical diagnosed LHON patients included.The mutation of G11778A was found in 36 cases,G3460A was found in 3 cases,and T14484C was found in 5 eases.Twelve cases of clinical diagnosed LHON patients didn't harbor any of the three common primary mutations,one case harbored the mutation C4171A,the mutation of G11696A was not found.There were totally 25 cases in the control group,the mutations of G11696A and C4171A were not found.Conclusion C4171A may be rare primary mt-DNA mutation in LHON patients in China.(Oph- thalmol CHN,2007,16:382-385)

AIMTo detect the changes of inducible nitric oxide synthase (iNOS) expression in liver following ischemia/reperfusion (I/R) of hindlimbs and to elucidate their significance.

METHODSI/R was established using the occlusion of the femoral arteries for 4 h and reopening for 2-24 h in rats. The expression of iNOS mRNA, and iNOS protein and the nitrotyrosine (NT), a marker of peroxynitrite (ONOO-), in liver tissue were detected with reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical technique, respectively. The liver superoxide dismutase (SOD) activity and malondialdehyde (MDA) contents were spectrophotometrically measured. The observation of pathologic changes of liver was made following the inhibition of iNOS by aminoguanidine (AG).

RESULTSCompared with control groups, the relative expression level of iNOS mRNA significantly increased in I/R group. There were more iNOS positive hepatocytes and more NT positive hepatocytes in I/R group than control groups. The contents of MDA markedly increased, while the activity of SOD significantly decreased in I/R group, compared with those in the control groups. The pathologic changes of rat liver became milder in I/R group following the inhibition of iNOS by AG.

CONCLUSIONThe expressions of iNOS mRNA and protein in liver are significantly upregulated, excess induction of iNOS-NO is contributed to the liver injury during the I/R of hindlimbs.

Animals ; Guanidines ; pharmacology ; Hindlimb ; blood supply ; Liver ; blood supply ; metabolism ; Male ; Malondialdehyde ; analysis ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; Superoxide Dismutase ; analysis

Objective: To study the dynamic changes of type Th22 cell immunological response during atherosclerosis process in experimental mice in order to provide a new theoretical basis for atherosclerosis therapy. Methods: 8 weeks C57BL/6J mice were divided into 2 groups: Experiment group,n=24 ApoE-/- mice and Control group, n=24 normal mice. All animals received high fat diet and the following indexes were compared between 2 groups at 0, 4, 8, 12 weeks after treatment: aortic atherosclerotic lesions were deifned by Oil red O staining, dynamic changes of Th22 cells in spleen were measured by lfow cytometry, mRNA expressions of interleukin-22 (IL-22), IL-22R1, AhR and T-bet in aorta were examined by RT-PCR, blood levels of IL-22 was detected by ELISA. Results: Compared with Control group, Experiment group had the increased area of aortic atherosclerosis (the ratio of plaque area/lumen area) and Th22 cell (CD4+ IL-22+/CD4+T cell) amount, elevated mRNA expressions of IL-22, IL-22R1, AHR, T-bet in aorta and higher blood levels of IL-22 at all time points, the differences between each time point (except 0 week) had the statistic meaning,P<0.05. In Experiment group, the differences between 2 adjacent time points, for the area of aortic atherosclerosis and mRNA expressions of AHR, T-bet: 4 weeks vs 0 week, 8 weeks vs 4 weeks, 12 weeks vs 8 weeks all had statistic meaning; for Th22 cell amount: 4 weeks vs 0 week, 8 weeks vs 4 weeks had statistic meaning and 12 weeks vs 8 weeks had no real distinction; for mRNA expressions of IL-22, IL-22R1 and blood levels of IL-22: 4 weeks vs 0 week had statistic meaning and 8 weeks vs 4 weeks, 12 weeks vs 8 weeks had no real distinctions. Conclusion: Hyperactive immunological response of Th22 cells might be involved in atherosclerosis process, the relevant mechanism should be further studied.

OBJECTIVESTo test the hypothesis that urine N-acetyl-beta-D-glucosaminidase (NAG) is a nearly biomarker for acute kidney injury in patients with acute paraquat poisoning.

METHODSForty-four patients with paraquat intoxication and 40 age and gender-matched healthy control participants were recruited. The urine N-acetyl-beta-D-glucosaminidase was determined by spectrophotometric methods.

RESULTSThe urine N-acetyl-beta-D-glucosaminidase activities in the patients with paraquat poisoning were higher than the corresponding values in the control participants (P<0.01); The prevalence rate of mortality was significantly higher in subjects with N-acetyl-beta-D-glucosaminidase activities ≥25 U/g Cr than in those N-acetyl-beta-D-glucosaminidase activities <25 Ulg Cr (34.4% vs 16.7%, P<0.01).

CONCLUSIONSThe urine N-acetyl-beta-D-glucosaminidase could be used as an early biomarker for acute kidney injury and predictor of mortality inpatients with acute paraquat intoxication.

Acetylglucosaminidase ; urine ; Acute Kidney Injury ; chemically induced ; diagnosis ; Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Paraquat ; poisoning ; Young Adult

OBJECTIVE: To assess the effects of Jinlongshe Granules (JLSG) on tumor growth of gastric carcinoma. METHODS: Fifty nude mice orthotopically transplanted with MKN-45 human gastric cancer cells were divided into five groups: untreated group, 5-fluorouracil (5-FU)-treated group and high-, medium-, and low-dose JLSG-treated groups. Corresponding antitumor drugs were administered in each group except the untreated group. The antitumor effects in vivo were evaluated. Cell cycle distribution and apoptosis of MKN-45 human gastric cancer cells were determined by using flow cytometry (FCM) and Annexin V-fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) staining assay. The ultrastructure of MKN-45 gastric cancer cells was observed by transmission electron microscope. RESULTS: In the mice treated with high-, medium-, and low-dose JLSG, the growth inhibition rates of gastric cancer were 68.13%, 55.94% and 50.31% respectively, and this antitumor effect was dose-dependent. In the mice treated with intraperitoneal injection of 5-FU, the growth inhibition rate of gastric cancer was 53.43% and not much different from those treated with JLSG. The apoptotic rates in the high-, medium-, and low-dose JLSG-treated groups were 22.81%, 28.27% and 38.54% respectively, in a dose-dependent manner, with the cell cycle arrested at G(0)/G(1) phase. An Annexin V-FITC/PI staining assay revealed that the percentages of early apoptotic cells in the three dose JLSG-treated groups were all significantly higher than that in the 5-FU-treated group, whereas the late apoptotic and necrotic cells were much more in the 5-FU-treated group than those in the three dose JLSG-treated groups. CONCLUSION: Jinlongshe Granules exert an inhibiting effect on MKN-45 human gastric cancer cell.

Objective To explore the mechanism by which the anti-human P185erbB2 scFv-Fc-IL-2(HFI)modulates tumor surface molecules and activates immune effector cells in vitro. MethodsMTT assay was used to test the proliferation and the LAK-like cytotoxicity. Flow cytometry assay was used to test the expression of ICAM-1, Fas and erbB2 receptors in tumor cells and the expression levels of CD molecules FasL and LFA-1 in human PBMC. Antibody-dependent cell-mediated cytotoxicity(ADCC)mediated by HFI against SKOV3, MCF-7 and SGC-7901 tumor cells was explored hv LDH release assay. Results The expression levels of ICAM-1 and Fas on SKOV3 cell treated with HFI were upregulated, from 24.85％ and 0.53％ to 85.36％ and 59.19％ respectively, while the expression levels of erbB2 on SKOV3, MCF-7 and SGC-7901 tumor cells treated with HFI were downregulated, from 98.48％, 42.60％ and 36.66％ to 94.01％,30.95％ and 12.36％ respectively. HFI could significantly enhance the proliferation activity of human PBMC, and CD3+ CD8+ T cells and CD3- CD16+ CD56+ NK cells were elevated, from 24.37％ and 6.90％ to 38.80％ and 13.45％ respectively. The expression levels of CD25, LFA-1 and FasL were significantly enhanced from 3.99％, 86.52％ and 5.02％ to 12.96％, 99.06％ and 16.19％. The LAK-like cytotoxicity of human PBMC treated with HFI against SKOV3, MCF-7,SGC-7901 tumor cells was significantly improved:HFI was effective in mediating ADCC against SKOV3,MCF-7 and SGC-7901 tumor cells which expressed high,medium and low levels of erbB2,respectively,and HFI-induced ADCC was correlated with the degrees of erbB2 expression on the tumor cells. Conclusion The expression levels of ICAM-1 and Fas on SKOV3 cell treated with HFI are significantly upregulated. The expression levels of erbB2 on SKOV3, MCF-7 and SGC-7901 tumor cells treated with HFI are downregulated. HFI can significantly enhance the proliferation activity of human PBMC. The LAK-like eytotoxicity of human PBMC treated with HFI against tumor cells is significantly enhanced. HFI iS effective in mediating ADCC and the activity of HFI-induced ADCC is correlated with the degrees of erbB2 expression on the tumor cells.

Objective To investigate the effects of rotating pulsed magnetic field on platelet derived growth factor a (PDGF-A) expression in periodontal tissue during tooth movement. Methods 30 white rabbits were random divided into 6 groups with 5 rabbits each group,including groups of 1,3,5,7,14 and 21 days. Under anesthesia condition by 2% pentobarbital sodium,the stainless coil springs were fixed between the first maxillary molar and the incisor producing the force of 80g. The experimental group was treated by rotating pulsed magnetic field and the force, the control group was only treated by t he force. The expression of PDGF-A was half-quantitatively investigated through immunohistochemical analysis. Results The expression of PDGF-AA in the experimental group enhanced apparently compared with that in the control group. There were significant differences among the 5,7, and 14day groups (5. 28 ± 0. 14 vs 2. 03 ±0. 18,7.63±0.27 vs 2. 84 ±0. 12,3.52 ±0. 16 vs 1.65 ±0.03;8.10±0.13 vs 4. 30 ±0. 21,13. 27 ±0. 31 vs 6.47 ± 0.15,5.66 ± 0.22 vs 3. 15 ± 0. 27, P ＜ 0. 05). The expression of PDGF-AA of 7 day group was higher than that of other groups. Conclusion Rotating pulsed magnetic field promotes the expression of PDGF-A in the periodontal tissue and alveolar bone remodeling.