Objective To determine if there is an early developmental resistance to seizure-induced hi ppocampal damage. Methods Five daily pentylenetetrazol-indu ced convulsions in immature rats beginning at postnatal day P10,P60 groups.In b oth groups, the latency of seizure, the latency of Ⅳ/Ⅴ grade, the lasting time of seizure and mortality of rats after seizure were used to measure sensitivity of seizure or the resistance to brain damage. Conventional histopathological me thod was utilized to observe morpbological changes and cell counting of dentate granule cells, CA 3,CA 1 and hilar neurnns. Timm histochemical technique was a dopted to study mossy fiber sprou- ting.Results 1.In the both groups(P10,P60),there were significant differences in the latency of seizure (1.07?0.55 vs 8.27?1.48 P

Objective: To evaluate the effect of bioactive peptides combined with probiotics on serum uric acid (SUA) in patients with hyperuricemia (HUA), so as to provide the evidence for prevention and treatment of HUA. Methods: The patients with HUA aged 18 to 65 years were selected and randomly divided into an intervention group and a control group. The patients in the intervention group received bioactive peptides combined with probiotics for 28 days at a dose of 3 g/d, while the patients in the control group received an equal dose of placebos. Demographic information, body mass index (BMI), blood pressure and blood lipid were collected through questionnaire surveys, physical examination and laboratory tests. SUA levels were detected before and after 14 days and 28 days of interventions. The differences of SUA levels between the two groups were compared using generalized estimation equation. Results: Totally 108 patients with HUA were recruited, including 54 patients in the intervention group and 53 patients in the control group (1 dropout). Before interventions, there were no statistically significant differences in gender, age, course of HUA, exercise duration, frequency of alcohol consumption, frequency of meat broth consumption, BMI, prevalence of hypertension and prevalence of dyslipidemia between the two groups (all P>0.05). After 14 days of interventions, the SUA levels of the patients in the intervention group decreased by 3.00 μmol/L, while those in the control group increased by 7.00 μmol/L. After 28 days of interventions, the SUA levels of the patients in the intervention group and the control group decreased by 26.00 μmol/L and 16.00 μmol/L, respectively. However, there was no statistically significant interaction between the intervention time and group (both P>0.05). Subgroup analysis showed that after 28 days of interventions, the decrease in SUA levels in the patients aged 55 years and older and without hypertension in the intervention group was greater than those in the control group (both P<0.05). Conclusions Bioactive peptides combined with probiotics showed no significant difference in reducing SUA levels in patients with HUA compared to the control group. The effect was more significant for patients aged 55 years and older and without hypertension.

OBJECTIVETo study the effect of Chinese drugs for invigorating qi and tonifying Shen (IQTS) on expression of CD4+CD25+ regulatory T cells in spleen and maternal-fetal interface of abortion-prone mice during pregnancy.

METHODSCBA female mice were mated with DBA/2 male mice to establish abortion-prone models, which were randomly divided into 4 groups, the negative control group (fed with normal saline), the positive control group (treated with CsA), the Chinese medicine group (treated with IQTS), and the Chinese and Western medicine group (treated with IQTS+CsA). Mice were sacrificed in batches on the 9th and the 14th day of gestation, their splenic and decidual tissues were taken out to analyse CD4+CD25+ regulatory T-cell expression by flow cytometry.

RESULTSCompared with the negative control group, the expression of splenic CD4+CD25+ regulatory T all significantly increased on the 9th day of gestation (P < 0.01 or P < 0.05). There was no statistical difference in intergroup comparison of the three treatment groups (P > 0.05). Compared with the negative control group, the expression of splenic CD4+CD25+ regulatory T all significantly increased on the 14th day of gestation (P < 0.01 or P < 0.05). Of them, its expression was the highest in the Chinese and Western medicine group, showing significant difference from that in the Chinese medicine group and the positive group (P < 0.01). The difference between the Chinese medicine group and the positive group was insignificant (P > 0.05). On day 9 of gestation, compared with the negative control group, the expressions of CD4+CD25+ regulatory T in maternal-fetal interface increased in the three treated groups, showing no statistical significance (P > 0.05). Its expression was ordered from high to low in sequence as the Chinese and Western medicine group, the positive control group, the Chinese medicine group, and the negative control group. On day 14 its expression was obviously enhanced in the Chinese and Western medicine group, showing statistical difference from that in the negative control group (P < 0.05). But its expression was obviously enhanced in the Chinese medicine group and the positive group, showing insignificant difference from that in the negative group. The same sequence was found in the percentage of CD4+CD25+ T cells in CD4+ T cells.

CONCLUSIONSChinese drugs for IQTS could up-regulate the expression of CD4+CD25+ regulatory T in spleen of abortion-prone mice in the early and late pregnancy stages. When combined with CsA, it also could up-regulate its expression in maternal-fetal interface in the mid and late pregnancy stages, suggesting that Chinese drugs for IQTS are facilitate to maintain the immune tolerance state in mice during pregnancy.

Abortion, Spontaneous ; drug therapy ; immunology ; metabolism ; Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Male ; Mice ; Mice, Inbred CBA ; Phytotherapy ; Pregnancy ; Spleen ; cytology ; drug effects ; T-Lymphocytes, Regulatory ; cytology ; drug effects ; metabolism

OBJECTIVE: To develop a multiplex allele-specific PCR (AS-PCR) assay with three-color fluorescence labeling for mitochondrial DNA (mtDNA) SNP typing. METHODS: Based on the principle of AS-PCR, the primer sets were designed for 20 SNP located on the coding region of mtDNA and divided into 2 groups labeled with FAM and HEX fluorescence, respectively. A primer set included two forward (reverse) allelic specific primers with different sizes and a generic reverse (forward) primer. Blood samples from 200 unrelated individuals were analyzed by AS-PCR and capillary electrophoresis. Three random samples at least for each SNP site were examined and verified by direct sequencing. The haplotype frequency was investigated. RESULTS: Distinct electropherograms of 200 blood samples were obtained successfully. The typing results of direct sequencing were identical to those obtained from AS-PCR. The minimum detectable DNA concentration was 0.2 pg under the system of 10 microL. The sensitivity of the DNA concentrations ranged from 0.5 to 5 pg. The 200 individuals were assigned into 15 haplotype, and the haplotype diversity was 0.906 0. CONCLUSION AS-PCR is a simple, rapid and efficient method for mtDNA SNP typing, and can be applied to forensic practice.
Alleles ; DNA ; DNA Primers ; DNA, Mitochondrial/analysis* ; Electrophoresis, Capillary ; Haplotypes ; Humans ; Mitochondria ; Polymerase Chain Reaction/methods* ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA

OBJECTIVETo observe the histopathological changes in rat hippocampus at different maturational stages after repeated kindled seizures, and to explore their underlying epileptogenesis processes.

METHODSThree groups of Wistar rats (postnatal days: P10, P20, P60) were given pentylenetetrazol (PTZ) intraperitoneal injection for 5 days to induce repeated kindled seizures, and the age-matched rats in control group were injected with normal saline. The behavioral changes, the morphology and the neurons counting in hippocampus, as well as the expression of NF-kappaB were observed.

RESULTS(1) In the three groups, the latency of seizure and the latency of IV/V grade were significantly lower in the rats of group P10 and P20 [(1.2 +/- 0.6) min and (14.4 +/- 2.3) min vs. (4.7 +/- 1.6) min and (24.5 +/- 4.5) min] than group P60 [(8.6 +/- 2.0) min and (41.9 +/- 4.5) min], whereas the duration of convulsion in group P10 and P20 [(46.2 +/- 4.8) min and (29.8 +/- 5.9) min] was longer than those of group P60 [(17.1 +/- 5.0) min]. (2) The neuron counting of CA(1), CA(3) and hilar in the P10 and P20 groups showed no differences as compared to their controls, whereas adult rats (P60) had a significant neuron loss in CA(1) and CA(3) pyramidal cells, compared with the control group [(6.3 +/- 1.5)/250 microm(2), (3.6 +/- 1.4)/250 microm(2) vs. (8.2 +/- 1.9)/250 microm(2), (5.6 +/- 1.7)/250 microm(2)]. However, the dentate granule cells in immature rats (P10) with daily seizures had a significant increase as compared with the controls [(23.3 +/- 3.1)/250 microm(2) vs. (16.3 +/- 1.6)/250 microm(2)]. (3) Prominent sprouting was seen in the CA(3) stratum pyramidal layer in all experimental rats with 5 daily seizures, regardless of the age. But the degree of sprouting had significant differences among the experimental groups (P < 0.05). (4) NF-kappaB was expressed significantly in CA(3), CA(1) and dentate granule cells 24 hours after PTZ-kindling when compared with the control groups, with the spectral density decreased with age.

CONCLUSION(1) There were great differences in the vulnerability to the repeated seizure-induced brain damage at different maturational stages in rats. The immature brain appeared to be less vulnerable to the repeated seizures. (2) There was less hippocampus neuron loss and milder mossy fiber sprouting after repeated seizures in the developing rats than mature ones, which may be a pathological evidence underlying the prospect that the immature brain was more resistant to the seizure-induced neuronal injury. (3) The high expression of NF-kappaB may exert a certain biological effects in the seizure-induced neuronal injury.

Age Factors ; Animals ; Hippocampus ; drug effects ; pathology ; NF-kappa B ; metabolism ; Pentylenetetrazole ; adverse effects ; Rats ; Rats, Wistar ; Seizures ; chemically induced

BACKGROUNDPulmonary embolism (PE) can be difficult to diagnose in elderly patients because of the coexistent diseases and the combination of drugs that they have taken. We aimed to compare the clinical diagnostic values of the Wells score, the revised Geneva score and each of them combined with D-dimer for suspected PE in elderly patients.

METHODSThree hundred and thirty-six patients who were admitted for suspected PE were enrolled retrospectively and divided into two groups based on age (≥65 or <65 years old). The Wells and revised Geneva scores were applied to evaluate the clinical probability of PE, and the positive predictive values of both scores were calculated using computed tomography pulmonary arteriography as a gold standard; overall accuracy was evaluated by the area under the curve (AUC) of receiver operator characteristic curve; the negative predictive values of D-dimer, the Wells score combined with D-dimer, and the revised Geneva score combined with D-dimer were calculated.

RESULTSNinety-six cases (28.6%) were definitely diagnosed as PE among 336 cases, among them 56 cases (58.3%) were ≥65 years old. The positive predictive values of Wells and revised Geneva scores were 65.8% and 32.4%, respectively (P < 0.05) in the elderly patients; the AUC for the Wells score and the revised Geneva score in elderly was 0.682 (95% confidence interval [CI]: 0.612-0.746) and 0.655 (95% CI: 0.584-0.722), respectively (P = 0.389). The negative predictive values of D-dimer, the Wells score combined with D-dimer, and the revised Geneva score combined with D-dimer were 93.7%, 100%, and 100% in the elderly, respectively.

CONCLUSIONSThe diagnostic value of the Wells score was higher than the revised Geneva score for the elderly cases with suspected PE. The combination of either the Wells score or the revised Geneva score with a normal D-dimer concentration is a safe strategy to rule out PE.

Aged ; Aged, 80 and over ; Angiography ; Female ; Fibrin Fibrinogen Degradation Products ; metabolism ; Humans ; Male ; Pulmonary Embolism ; diagnosis ; diagnostic imaging ; metabolism ; Retrospective Studies

Objective To evaluate the micropermeability on bonding hydrophobic adhesive to dentin with ethanol-wet bonding under simulated pulp pressure.Methods Twenty-four intact human third molars were used in the study.After the enamel of occlusal surfaces was removed,the molars were randomly divided into six groups.Adper Scotchbond Multi-Purpose was used in the control group; in the experimental groups,the dentin surfaces were saturated with ethanol for 20 s ( group 1 ),1 min ( group 2 ),2 min ( group 3 ),3 min ( group 4 ) or with a series of increasing ethanol concentrations before application of hydrophobic adhesive ( group 5 ).All the bonding procedures were done under simulated pulp pressure.After 24 hours,micro-tensile bond strength test were performed on the specimens.Bonding interfaces were observed under laser scanning confocal microscope (LSCM) after the pulp chamber were filled with a water-soluble fluoroprobe rhodamine B for 3 hours.Results Compared with the control group[(38.14 ± 4.97 ) MPa],bond strengths in group 1 [(21.02 ± 7.23 ) MPa]and group 2 [( 29.64 ± 3.81 ) MPa]were statistically lower ( P ＞ 0.05 ),while bond strngth in group 3 [( 38.40 ± 5.03 ) MPa],group 4 [( 37.26 ± 4.68 ) MPa]and group 5[(40.12 ±5.95) MPa]were similar to the control group (P＜0.05).The images taken by LSCM showed that with extension of ethanol-wet time,the deposition of fluorescent dye in hybrid layer and along the dentinal tubules decreased gradually.Especially in group 5,only spare fluorescent dye deposition could be detected in the hybrid layer.Conclusions Dentin saturated with ethanol for more than 2 min before bonding hydrophobic adhesive to dentin could provide favorable bond strength and decreased the micropermeability of bonding interfaces under simulated pulp pressure.

OBJECTIVETo construct a directional differentiation model from mouse embryonic stem cells into leydig-like cells in vitro.

METHODSMouse ES-D3 cells were transfected with plasmid containing steroidogenic factor 1 (SF-1) gene, then treated with RA and 8Br-cAMP, while the cells transfected with empty plasmid were used as the negative controls. The morphology of leydig-like cells differentiated from ES-D3 cells was observed with light microscopy. The expression levels of StAR, P450scc and 3β-HSD were detected by RT-PCR, Western Blot and fluorescence microscopy analysis in leydig-like cells derived from the ES cells.

RESULTSES-D3 cells were transfected with plasmid containing SF-1 gene successfully, and SF-1 was expressed 24 h after transfection. The SF-1-transfected ES-D3 cells were induced by RA and 8Br-cAMP to differentiate into leydig-like cells. The differentiated cells showed spindle shape with tentacles, which expressed the specific protein marker for leydig cells 3β-HSD1 and P450scc. Meanwhile, in these leydig-like cells, the expression of StAR increased compared with control group. 3β-HSD1, P450scc and StAR were not detected in negative control group.

CONCLUSIONWhen the ES-D3 cells are transfected with SF-1 plasmid and then treated with RA and 8Br-cAMP, the cells are able to differentiate into leydig-like cells, indicating that the model of directional differentiation of ES cells into leydig-like cells has been constructed successfully.

Animals ; Cell Differentiation ; drug effects ; genetics ; Cell Line ; Embryonic Stem Cells ; cytology ; metabolism ; Leydig Cells ; cytology ; metabolism ; Male ; Mice ; Steroidogenic Factor 1 ; genetics ; Transfection

Previous studies have demonstrated the effects of brain-derived neurotrophic factor (BDNF) on promoting proliferation of multiple myeloma (MM) cells and inducing angiogenesis in MM in vitro. This study was aimed to further explore whether BDNF/TrkB pathway is a potential therapeutic target in MM, and to elucidate the advantages and disadvantages of two ways developed for human myeloma xenograft in animal models. The models of xenograft tumors were established in the non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice by subcutaneous or intravenous injection of human myeloma cell line RPMI8226. Mice were monitored daily for life state, and the volume of subcutaneous tumors were measured after inoculation. 3 weeks after inoculation, red blood cell counts, BDNF level in plasma, human lambda light chain and calcium level in serum of NOD/SCID were detected every two weeks. The histological and cytological examinations were performed to observe pathological features of tumors. Using flow cytometry to observe the expression of human CD38+ cell in murine blood and bone marrow. The changes of bone density and skeletal lesions were detected by computer radiography. The results showed that the subcutaneously injected animal model showed a high growth efficiency of RPMI8226 subcutaneous tumors (5/5) and several pathological features of plasmacytomas. There were neither obvious increase in lambda light chain and calcium levels, nor spread of human MM cells to murine bone marrow and no radiological evidence of skeletal lesions. The intravenously injected animal model had relative low efficiency for growth of tumors (4/7) but MM cells could engraft and proliferate in murine bone marrow. The human lambda light chain could be detected in serum as early as 3 weeks after inoculation. Myeloma-bearing mice had high level of lambda light chain and high calcium in serum and resorption of the murine bone. Furthermore, the concentrations of BDNF were increased with the tumor growth in both models with (73 +/- 11) pg/ml and (105 +/- 18) pg/ml in plasma respectively at 9 weeks after inoculation. It is concluded that two appropriate MM xenograft NOD/SCID animal models were established, both of which show high BDNF levels in the plasma. Therefore, two valuable in vivo systems to explore novel therapeutic target (BDNF/TrkB) in MM have been set up successfully.
Animals ; Brain-Derived Neurotrophic Factor ; metabolism ; Disease Models, Animal ; Male ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Multiple Myeloma ; metabolism ; Neoplasm Transplantation

OBJECTIVETo compare the pituitary down-regulatory effects of the two gonadotropin-releasing hormone agonists Alarelin and Triptorelin in the long protocol of ovulation induction in in vitro fertilization and embryo transfer (IVF-ET).

METHODSWe included in this study 122 patients aged 24-39 years treated by IVF-ET for secondary infertility, with 10-20 pre-antral follicles and obstruction of the fallopian tube. Seventy-eight of them received Alarelin, and the other 44 Triptorelin. Comparative analyses were made on the pituitary down-regulatory effects of the two gonadotropin-releasing hormone agonists and the clinical outcomes of IVF-ET.

RESULTSNo premature LH surge and ovulation, nor severe hyperovarian stimulation syndrome was found in either group. There were no significant differences between the two groups in the mean dose and duration of gonodatropin treatment, the numbers of oocytes retrieved, mature oocytes and top-quality embryos, and the rates of 2PN, multi-sperm fertilization, cleavage, embryo transfer, embryo implantation, clinical pregnancy and early miscarriage (P > 0.05), but the rate of cancelled cycles was significantly higher in the Triptorelin than in the Alarelin group (P < 0.05).

CONCLUSIONAlarelin and Triptorelin can achieve similar pituitary down-regulatory effects and clinical outcomes in IVF-ET when used in the long protocol of ovulation induction.

Adult ; Embryo Transfer ; methods ; Female ; Fertilization in Vitro ; methods ; Gonadotropin-Releasing Hormone ; agonists ; analogs & derivatives ; pharmacology ; Humans ; Infertility, Female ; therapy ; Ovulation Induction ; methods ; Pituitary Gland ; drug effects ; Triptorelin Pamoate ; pharmacology