Aged ; Altitude ; Endothelin-1 ; blood ; Female ; Humans ; Hypertension, Pulmonary ; Male ; Middle Aged ; Pulmonary Heart Disease ; physiopathology ; Ventricular Function, Right

Anemia, Macrocytic ; Chromosome Deletion ; Chromosomes, Human, Pair 5 ; Cri-du-Chat Syndrome ; Humans ; Trisomy

Objective Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD, OMIM 605814 ) is a novel autosomal recessive disease results from mutations in the gene SLC25A13 that encodes for citrin, a liver-type aspartate/glutamate cartier located in the mitochondrial inner membrane. Most of the Chinese NICCD patients diagnosed by genetic analysis had the sameSLC25A13 mutations as Japanese, however, in some cases, the known mutations were not detected. This research aimed to identify novel SLC25A13 mutations in Chinese NICCD patients and to explore the experimental conditions for their genetic diagnosis.Methods Genomic DNA was extracted from blood samples of 3 NICCD patients from Taiwan (P757), Guangdong (P1194) and Hebei (P1443) Province of China, respectively; and all the 18 exons and their flanking sequences of SLC25A13 gene were sequenced. Furthermore, the identified novel mutations were diagnosed by amplification with PCR, digestion with corresponding restriction endonuclease, and agarose gel electrophoresis.Results Three novel mutations identified in SLC25A13 gene of the 3 NICCD patients were an abnormal splicing IVS7-2A>G (P757), a missense A541D (c. 1622C > A, P1194) and a nonsense R319X (c. 955C > T, P1443). The PCR-RFLP procedures for their genetic diagnosis were also established, with specific fragments on electrophoresis after digestion of the PCR products with three different restriction endonucleases Msp Ⅰ, Hpy188Ⅰ and Taq Ⅰ, respectively.Conclusions The three novel mutations in SLC25A13 gene of Chinese NICCD patients were first identified, suggesting that SLC25A13 mutation distributed in Chinese population is somewhat different from that in Japanese. Moreover, the PCR-RFLP diagnostic procedures established in this research provide valuable tools not only for the genetic diagnosis of NICCD but also for further molecular epidemiologic investigations in Chinese population.Acknowledgement We are grateful to all research subjects and their family members for their cooperation, and to many members of medical staff who contributed much to this research. This study was financially supported partially by Guangdong Provincial Research Grant for Science and Technology (No. 2004B50301008) and the Major State Basic Research Development Program of China (No. 2007CB511901 ), and by Grants-in-Aid for Scientific Research (B: Nos. 16390100 & 19390096 ) and for Asia-Africa Scientific Platform Program (AASPP) from the Japan Society for the Promotion of Science.

Antigens, CD20 ; metabolism ; Epstein-Barr Virus Infections ; virology ; Female ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Lymphoma, Large B-Cell, Diffuse ; pathology ; virology ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; virology

Hepatitis B virus ; Hepatitis B, Chronic ; immunology ; virology ; Humans ; T-Lymphocytes ; immunology

Objective To isolate and identify the adult neural stem cells from the parenchyma of spinal cord in adult mouse.Methods The parenchymal spinal cord from adult mouse was dissected and dissociated by mechanical trituration.The tissue suspension was cultured in serum-free DMEM/F12 medium supplemented with EGF and B27.The cell colonies generated from a single cell were screened by limited dilution and incubated with BrdU.The cell colonies were transferred into medium with serum to induce differentiation.The cells were identified with antibodies to Nestin,BrdU,MAP2 and GFAP by immunofluorescence staining.Results The cells were cultured for seven days to generate proliferative neurospheres.The majority of cells in these neurospheres expressed Nestin and were differentiated into MAP2-positive cells and GFAP-positive cells in medium containing with fetal bovine serum.Conclusion A significant number of neural stem cells are present in the parenchymal adult mouse spinal cord and can proliferate and also give rise to neurons and glia in vitro.

Cryptococcosis ; etiology ; pathology ; Glycogen Storage Disease Type I ; complications ; pathology ; Humans ; Infant ; Male ; Meningitis, Fungal ; etiology ; pathology

OBJECTIVETo investigate the effect of omega-3 polyunsaturated fatty acids(omega-3PUFAs) on the apoptosis of human gastric cancer cell line SGC-7901 and to explore the potential mechanisms.

METHODSCells were treated with eicosapentaenoic acid (20:5 omega-3,EPA) or docosahexaenoic acid (22:6 omega-3, DHA) at concentrations of 10, 20 and 40 microg/ml. Cell growth and apoptosis were analyzed with MTT assay, cell morphology, DNA electrophoresis and flow cytometry. Mitochondrial membrane potential ( triangle right psi mt) was measured by fluorescent probe rhodamine 123. The distribution of cytochrome C in mitochondria and cytosol was determined by enzyme-linked immunoadsorbent assay. The composition of mitochondrial membrane phospholipid(MMP)was examined by gas chromatography.

RESULTSBoth EPA and DHA markedly inhibited the SGC-7901 cell growth and induced apoptosis in a time- and dose-dependent manner. After incubation of the cells with 40 microg/ml EPA or DHA for 24 hours, the level of Deltapsimt siginificantly decreased (P<0.001), and cytochrome C largely released into cytosol from mitochondria. The proportions of EPA and DHA in MMP rapidly elevated while that of arachidonic acid sharply decreased.

CONCLUSIONSomega-3PUFAs inhibit the growth of gastric cancer cells through promoting apoptosis. Compositional and functional alterations in mitochondrial membrane may be an important initiator of apoptosis induced by omega-3PUFAs.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cytochromes c ; metabolism ; Fatty Acids, Omega-3 ; pharmacology ; Humans ; Membrane Potential, Mitochondrial ; Mitochondria ; metabolism ; pathology ; Stomach Neoplasms ; metabolism

OBJECTIVETo observe clinical results of Chinese herbs promoting blood circulation and removing blood stasis on the treatment of reflex sympathetic dystrophy (RSD) with type of stagnation of vital energy and blood stasis.

METHODSRSD with type of stagnation of vital energy and blood stasis was distinguished as erubescence, high temperature, perspiration, damp and acro-edema, with middle level pain. From 2006 to 2008, 58 patients with RSD of stagnation of vital energy and blood stasis were randomly divided into the treatment group (30 cases) and the control group (28 cases). The former were treated with Chinese medicine to activate blood circulation and improve bone and muscle nourishment. Chinese medicine includes: Caesalpinia Sappan 10 g, Ligusticum Chuanxiong 6 g, Frankincense 6 g, Angelica 10 g, Safflower 6 g, Myrrh 6 g, Ground Beetle 10 g, Araliaceae 3 g, Radix Paeoniae Rubra 10 g, Pericarpium Citri Reticulatae 5 g, Lawn Pennywort Herb 15 g, Manis Pentadactyla 10 g, Corydalis Yanhusuo 10 g, Rhizoma Drynariae 15 g, which were boiled into decoction and the patients were take orally everyday with a course of treatment for 10 days, together with the boiled Chinese traditional medicine of stretching muscle and activating blood circulation to fume and wash the limbs twice everyday. The compatibility of medicines in prescription includes: Lycopodium Japanicum Grass 10 g, Gentiana Macrophylla Pall 10 g, Radix Angelicae Pubescentis 10 g, Angelica 10 g, Uncaria 10 g, Frankincense 6 g, Myrrh 6 g, Safflower 6 g. Control group were treated with a placebo of the same color for oral use and external application. The delivery times, method and the time of therapy were all the same as the treatment group. After 30 days' treatment, the effective indexes of VAS pain score and swelling condition were observed in both groups.

RESULTSVAS pain score: the treatment group decreased (3.8 +/- 0.8) points and the control group decreased (1.0 +/- 0.3) points, the difference between the two groups was significantly (P < 0.01). There was significantly difference in volume decrease of the swelling limb between treatment group (21.8 +/- 2.5) ml and the control group (10.3 +/- 2.1) ml (P < 0.01). The efficiency difference between treatment group and control group was significantly(P<0.01).

CONCLUSIONWith the different treatment based on different syndrome and emphasis on the nourishment of bone and soft tissue, treated by Chinese medicine to promote blood circulation and remove blood stasis in stagnation of vital energy and blood stasis, RSD get a favorable result.

Adult ; Aged ; Blood Circulation ; drug effects ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Reflex Sympathetic Dystrophy ; drug therapy ; Treatment Outcome

Objective To analyze the outcome of surveillance results on plague and to provide the evidences for the policy making in Longlin county Guangxi. Methods The epidemic data and the surveillance results of plague were analyzed and assessed with epidemiology methods in Longlin county Guangxi from 2000 to 2009, and the density of rodents, the rodents infected with flea, flea index and other indicators were calculated. Regional composition of the rats and fleas were analyzed. Results A totally of 4829 rats were captured and 4737 fleas were collected in the past 10 years, Rattus Flavipestus(81.92%,3956/4829) and Xenopsylla Cheopis (79.04%,3744/4737) were dominant species. The annual average density of rodents, the rodents infected with flea, index of flea were 3.30%(4829/146 206), 27.99%(1351/4827) and 0.98(4737/4827), respectively. A totally of 4792 rats were examined and 10 strains Yersinia Pestis were isolated. Indirect hemorrhagic assessed(IHA) was used to test the F1 antibody against plague in the blood serum of the rats and indicator animals, and 3 positive rats and 24 positive animals were found, respectively. Twenty seven natural villages in 3 towns had been involved in the plague. Conclusions The plague foci exists in Longlin county of Guangxi province. The plague foci in the areas have the same feature with the plague foci of Rattus Flavipectus. There is a potential risk for plague in this region, we should improve the quality of surveillance, increase indicator animals of the plague, and try to apply new surveillance method.