To observe the effect of intracarotid administration of endothelin (ET-1) on electrical activity of neurons within rostral ventrolateral medulla (RVLM) region, 87 spontaneous active units were extracellularly recorded in 35 Sprague-Dawley rats with sino-aortic denervation. The results obtained are as follows. (1) Intracarotid administration of ET-1 (0.3nmol/kg) increased the discharge firing rate from 17.8±1.5 to 20.9±1.4 spikes/s (P<0.01) in 30 out of 36 RVLM neurons, while blood pressure and heart rate had no significant change. (2) BQ-123 (0.67nmol/kg), a selective ETA blocker, completely blocked the facilitatory effects of ET-1 in 11 out of 14 units. (3) In 10 out of 11 units, glibenclamide (3.3 nmol/kg), a blocker of ATP-sensitive potassium channel, had no effect on the action of ET-1. (4) After ablation of area postrema (AP), the facilitatory action of intracarotid administration of ET-1 on 19 units of RVLM was abolished, while in 7 units of sham ablation animals the response of neurons to ET-1 remained unchanged. Taken together, intracarotid-administered ET-1 may act on the ETA receptors in neurons of AP, thereby resulting in the facilitating effect on RVLM neurons through the efferent projection of AP.

The effects of microinjection of adenosine (Ado) into area postrema (AP) on mean arterial pressure (MAP), heart rate (HR) and renal sympathetic nerve activity (RSNA) were examined in 53 anesthetized Sprague-Dawley rats. The results obtained are as follows. (1) Following microinjection of Ado (1 ng/60 nl) into AP, MAP, HR and RSNA were decreased from 13.76±0.46 kPa, 356.28±4.25 bpm and 100±0% to 11.23±0.49 kPa (P<0.001), 336.91±5.23 bpm (P<0.01) and 70.95±5.19% (P<0.001), respectively; (2) 8-phenyltheophylline (150 μg/kg, 0.2 ml,iv), a nonselective adenosine receptor antagonist, and 8-cyclopentyl-1,3-dipropylxanthine (500 μg/kg, 0.2 ml, iv), a selective A1 adenosine receptor antagonist, blocked the inhibitory effect of Ado completely; and (3) glibenclamide (5 mg/kg, 0.2 ml, iv), a blocker of ATP-sensitive potassium channel, also abolished the effect of Ado. The above results indicate that microinjection of Ado into AP induces inhibitory effects on MAP, HR and RSNA, which may be related to activation of ATP-sensitive potassium channels mediated by A1 receptors.

OBJECTIVETo determine the molecular basis of late onset ornithine transcarbamylase (OTC) deficiency in a Chinese family of Han nationality and the exon sequences of OTC gene of this patient.

METHODSPolymerase chain reaction-single strand conformation polymorphism and direct sequencing were used to identify the mutation type.

RESULTSA missense mutation E122G in the conserved residue of exon 4 was identified which is unreported before.

CONCLUSIONThe E122G mutation in human OTC gene may cause late onset OTC deficiency.

Age of Onset ; Base Sequence ; Child, Preschool ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Family Health ; Fatal Outcome ; Female ; Humans ; Male ; Models, Molecular ; Mutation, Missense ; Ornithine Carbamoyltransferase ; chemistry ; genetics ; Ornithine Carbamoyltransferase Deficiency Disease ; enzymology ; genetics ; pathology ; Pedigree ; Polymorphism, Single-Stranded Conformational ; Protein Structure, Secondary

To explore the difference of negative regulatory factors among T lymphocyte subsets in bone marrow (BM) of myelodysplastic syndromes (MDS) and their relations to apoptotic gene Fas, different lymphocyte subsets in BM were categorized by monoclonal antibodies with 3 color fluorescence using flow cytometry, and the intracellular cytokines such as tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) were determined following marrow cells culture. Then Fas mRNA of bone marrow mononuclear cells (BMMNC) were examined by RT-PCR. The results showed that TNF-alpha, IFN-gamma levels in BM of MDS both increased, the former produced by cells CD4+CD45RO+, CD8+CD45RO+, the latter by cells CD4+CD45RO+, CD8+CD45RO+, CD8+CD45RA+, in which the cells CD8+CD45RO+ were dominant. Fas mRNA expression had relationship with IFN-gamma produced by T cells but not with TNF-alpha. It is concluded that in hematopoietic microenvironment of MDS, not only the T lymphocyte subsets are in disorder, but also negative regulatory factors secreted by T lymphocyte increase. T lymphocytes play an important role in producing IFN-gamma in patients with MDS.
Adult ; Aged ; Bone Marrow Cells ; metabolism ; Female ; Humans ; Interferon-gamma ; biosynthesis ; Male ; Middle Aged ; Myelodysplastic Syndromes ; metabolism ; pathology ; RNA, Messenger ; analysis ; T-Lymphocyte Subsets ; metabolism ; Tumor Necrosis Factor-alpha ; biosynthesis ; fas Receptor ; genetics

Objective To study the neuronal apoptosis and expressions of apoptosis-related genes in the hippocampal CAI region after cerebral ischemia in gerbils and investigate the protective effects of mild hypothermia. Methods Seventy-two gerbils were randomized equally into sham-operated group, hypothermic sham-operated group, normothermic ischemia-reperfusion (IR) group and hypothermic IR groups, and 5-minute forebrain iscbemia was induced in the latter 2 groups by bilateral common carotid artery obstruction. Normothermic or mild hypothermic condition was applied to the groups after the operation as indicated. At 1, 3 and 7 days after the operation, 6 gerbils were selected from each group for behavioral test using open field test, followed by detection of neuronal apoptosis in the hippocampal CAI region using TUNEL staining and by immunohistocheraistry for p53 and nuclear factor-kB (NF-kB) expressions. Results In normothermic condition, the 5-minute forebrain ischemia induced significant enhancement of the exploratory activities in the gerbils 1, 3 and 7 days after the operation (P<0.05). This enhancement was observed only 1 day after the operation in mild hypothennic condition (P<0.05). Increased neuronal apoptosis and up-regulated expressions of p53 protein and NF-kBin the hippocampal CA1 region occurred after the forebrain ischemia as shown by TUNEL staining and immunohistochemistry, and all these changes were significantly inhibited by the application of mild hypothermia (P<0.05). Conclusions Up-regulated p53 and NF-kB protein expression in the hippocampal CA1 region might be one of the mechanisms for ischemia-induced neuronal apoptosis in gerbils, and mild hypothermia may produce protective effects against these changes for brain protection following the iscbemia.

OBJECTIVETo investigate the actual condition of the sexual physiological and psychological development of the high school students in Yibin in order to get a reliable basis for sexual education of the teenagers.

METHODSWith a proportion of 1% to the whole, 2,770 students were randomly selected from eight high schools in the urban and rural areas of the city. We devised a questionnaire and asked each student to fill in his or her answers presently.

RESULTSSo far as sexual physiological and psychological development was concerned, the high school students of Yibin were found rather precocious, with very little sexual knowledge and psychological endurance in sexual affairs and a relative lack of sexual education.

CONCLUSIONIt is imperative to extend the scope of puberty sexual education in high schools. Teenagers must be taught different kinds of sexual knowledge at different periods of growth as well as how to avoid sexually transmitted diseases and gestation. The sexual knowledge level of the teachers must also be raised. It is a must to establish service or specialist consultation hot lines about sexual knowledge for teenagers. Parents are expected to change their traditional views and assume an active role in the joint efforts of sexual education for their children.

Adolescent ; Child ; China ; Female ; Humans ; Male ; Menarche ; Psychology, Adolescent ; Psychosexual Development ; Rural Population ; Sexual Development ; Students ; psychology ; Surveys and Questionnaires

OBJECTIVETo develop a multiplex PCR protocol, which could be suitable for routine screening of microdeletions on the Y chromosome in azoospermic and oligozoospermic male infertility patients.

METHODSFive multiplex sets were established. Eighty-seven azoospermic and oligozoospermic patients undergoing intracytoplasmic sperm injection (ICSI) in the in vitro fertilization (IVF) center and 30 azoospermic men undergoing testicular biopsy in the clinic of Urology Surgery were screened for microdeletions of Y chromosome.

RESULTSA total of 19 (16.2%) cases of microdeletions were found in 117 azoospermic and oligozoospermic patients by screening of Y chromosome microdeletions. Of these, 11 cases (18.0%) were found in 61 oligozoospermic patients, and 8 cases (14.3%) were found in 56 azoospermic patients.

CONCLUSIONThe multiplex PCR protocol presented in this study is an easy-to-do and reliable method for detecting microdeletions on the Y chromosome. Routine screening of microdeletions on the Y chromosome for azoospermic and oligozoospermic patients is essential.

Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Female ; Genetic Testing ; methods ; Humans ; Infertility, Male ; diagnosis ; genetics ; Male ; Polymerase Chain Reaction

OBJECTIVETo explore cell culture techniques for amplification of oval cells with preservation simultaneously of the stem cell characteristics.

METHODSOval cell line OC3 was cultured in RPMI 1640 supplemented with 15% fetal bovine serum and 20 µg/L EGF. Cells were harvested every 5 passages and were examined with biomarkers including OV-6, c-kit, gamma-glutamyl transpeptidase, placental form of glutathione-S-transferase (GST-P), pyruvate kinase M₂, pyruvate kinase L and albumin using techniques including RT-PCR, immunocytochemistry, and enzymo-cytochemistry.

RESULTSOC3 cell lines could be amplified abundantly in-vitro associating with expression of infant liver cell markers at various level, including OV-6, c-kit, gamma-glutamyl transpeptidase, GST-P, pyruvate kinase M₂, but no expression of mature hepatocyte markers detected including pyruvate kinase L and albumin.

CONCLUSIONSAmplification of OC3 cells with preservation of the stem cell phenotype and high proliferation index can be achieved up to the 79(th) passages by culturing in RPMI 1640 supplemented with 15% fetal bovine serum and 20 µg/L EGF.

Animals ; Antigens, Differentiation ; metabolism ; Cell Culture Techniques ; Cell Differentiation ; Cell Line ; Culture Media ; Glutathione Transferase ; metabolism ; Hepatocytes ; cytology ; metabolism ; Liver ; cytology ; growth & development ; Phenotype ; Proto-Oncogene Proteins c-kit ; metabolism ; Pyruvate Kinase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; metabolism

OBJECTIVETo investigate the efficacy of imatinib in the treatment of chronic myeloid leukemia (CML) and analyse the treatment outcome and related factors.

METHODSNinety five CML patients were treated with imatinib in our hospital from May 2002 to May 2006. The outcomes and related factors were analysed.

RESULTS(1) One year after therapy, there were 95.5% of chronic phase (CP) patients achieved complete hematologic response (CHR). Fifty-two patients with complete cytogenetic dates were divided into primary-therapy group (n = 19) and secondary-therapy group (n = 33). The major cytogenetic responses (MCyR) at 6-, 12-, 18-, 24- and 30-months after therapy for the former group were 84.2%, 84.2%, 89.5%, 89.5% and 94.7%, and for the latter group were 36.4%, 39.4%, 39.4%, 39.4% and 39.4%, respectively (P < 0.01). The expected survival at 12-, 24-, 36- and 50-month after imatinib treatment for CP group was (98.1 +/-1.9)%, (87.8 +/- 7.1)%, (81.9 +/- 8.7)% and (81.9 +/- 8.7)%, respectively. (2) Twelve month after therapy, there are 70% of accelerated phase (AP) patients achieve CHR and 10% get MCyR. The expected survival at 12-, 24- and 36-month after imatinib treatment for AP group was (63.0 +/- 17.7)%, (15.8 +/- 14.3)% and (15.8 +/- 14.3)%, respectively. (3) Six month after therapy, 57.9% of blast crisis (BC) patients achieve CHR, with the expected survival at 12- and 24-month of (40.6 +/- 12.3)% and 0, respectively. (4) COX analysis CP group indicated that imatinib therapy administered for previously untreated was an independent favorable prognostic factor. Conclusion (1) Imatinib as a primary treatment for CP CML can significantly improve the survival time as compared with that AP or BC patients or with that used in previously treated patients. (2) Imatinib could induce hematologic, even cytogenetic response to a certain extent, in CP or BC patients and prolong the survival time.

Adolescent ; Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Benzamides ; Female ; Humans ; Imatinib Mesylate ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; Male ; Middle Aged ; Piperazines ; therapeutic use ; Pyrimidines ; therapeutic use ; Treatment Outcome ; Young Adult

OBJECTIVESTo observe the role of PPARgamma during the activation process of hepatic stellate cells (HSC).

METHODSBy morphology and RT-PCR, we study the changes of expression of PPARgamma in culture-activated HSC or in vivo activated HSC induced by dimethylnitrosamine (DMN).

RESULTSIn vitro, the expression level of PPARgamma in freshly isolated HSC (0.72+/-0.01) significantly reduced to 0.48+/-0.03 on the third day of culture (t = 19.8372, P<0.01), and reduced 70% on the seventh culture-day and could not be detected after the second passage. In vivo, HSC freshly isolated from normal control rats expressed PPARgamma (0.76+/-0.01). During the development of rat liver fibrosis induced by DMN, the expression level significantly reduced to 0.46+/-0.02 after the third injection of DMN (t = 29.5318, P<0.01), and reduced 66% on the end of first week and could not be detected on the end of second and third week.

CONCLUSIONThe expression of PPARgamma might play an important role on the maintenance of resting-form of HSC, and the reduction of expression of PPARgamma might be an early event during the activation process of HSC.

Animals ; Liver ; cytology ; Liver Cirrhosis ; etiology ; pathology ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Receptors, Cytoplasmic and Nuclear ; physiology ; Transcription Factors ; physiology