This study was aimed to discuss the distribution and protein expression level ofμ-opioid receptor (MOR) in hippocampus of premenstrual syndrome (PMS) liver-qi stagnation rat model, in order to initially reveal the action mechanism of PMS liver-qi stagnation and intervention effect ofShu-Yu (SY) capsule. Chronic restraint stress method was used to copy PMS liver-qi stagnation rat model.SY capsule ofTiao-Gan prescription was given as intervention. Immunofluorescence (IF) and western blot (WB) technique were used to detect MOR in hippocampal CA1 and CA3 brain area of rats from each group. The results showed that compared with the normal group, the hippocampus MOR distribution arrangement was messy with increased protein concentration in the model group (P< 0.01). After drug intervention, the MOR protein level returned to normal level. It was concluded that the pathogenesis of PMS liver-qi stagnation may be associated with high expression of MOR in hippocampus CA1 and CA3 region of rats. SY capsule can effectively correct and restore it to nearly normal level. It may be one of the central mechanisms in SY capsule treatment of PMS liver-qi stagnation.

AIM: To investigate the anti-cancer effect of cytotoxic T lymphocytes (CTL) activated by sensitized dendritic cells (DCs). METHODS: Immature DCs were induced in vitro from peripheral blood monocytic cells (PBMC) and sensitized by adding tumor cells antigen extract. DCs were identified by their morphology and surface markers. MTT assay was used to evaluate the killing activity of CTL activated by sensitized DCs. The effects of specific CTL cells on inhibiting transplanted tumor HT-29 growth and on preventing HT-29 tumor generation were evaluated by injecting CTL into nude mice. RESULTS: After cultured for seven days, a large number of activated DCs were obtained with typical morphology, extensive stimulatory proliferation capacity and high CD80 (63.5%), CD83 (67.6%) and CD3/HLA-DR (83.2%) expressions. The killing activity of CTL at 20∶1 ratio of effective cells to target cells was more than 75% to tumor cells, 35%-45% to homologous cell line and weaker to other germ cell line (P

Objective To evaluate the Carryover between the chemistry and immunoassay in Beckman Coulter Laboratory Au-tomation System and decide to whether sharing samples for testing between chemistry and immunoassay systems or not. Methods According to a certain order,high concentration samples and low concentration samples of HCG with different sample volume (500 μl,2 000 μl)were tested on Beckman AU5421 automatic biochemical analyzer.The HCG of low concen-tration samples were then tested to evaluate the carryover between the chemistry and immunoassay and explored the correc-tive procedure to deal with the carryover by increasing special cleaning process of beckman AU5421 automatic biochemistry analyzer.Results Under different sample volume,the carryover in a single module and as a whole of the beckman AU5421 automatic biochemistry analyzer were 5.44,15.47,23.51 and 45.96 ppm respectively (t=14.553,P <0.001;t=5.527,P =0.005;t=3.985,P =0.016;t=20.457,P <0.001).By increasing special cleaning process the carryover of 0.22 ppm was detected in 500 μl sample volume of the beckman AU5421 automatic biochemistry analyzer as a whole.Conclusion The car-ryover between the chemistry and immunoassay in Beckman Coulter Laboratory Automation System could been sovled by in-creasing special cleaning process of beckman AU5421 automatic biochemistry analyzer.

Calcium hydroxide (CH) is applied to improve disinfection of root canals in most root canal retreatment. This study aimed to analyze the CH removal efficacy using 7 different root preparing files (K file, pre-curved K file, EndoActivator, Ultrasonic file, pre-curved ultrasonic file, F file and needle irrigation alone) with apical transportation. Standardized models of curved canal with such apical transportation or not were set up before applying CH to root canal for 7 days. Seven techniques described above were used for its removal. Then the roots were disassembled and digital photos were taken. The ratio of residual CH in the overall canal surface was calculated using the image analyzer image pro plus 6.0. The data were analyzed using one-way ANOVA with post hoc Tukey test. Results revealed that CH was effectively removed (P<0.05) by using all 6 mechanical methods except irrigation alone. In curved root canals with apical transportation, EndoActivator, pre-curved ultrasonic file and F file were found to be more effective in removing CH than the other four file (P<0.001), while there was no significant difference among EndoActivator, pre-curved ultrasonic file and F file groups (P>0.05). The percentage of residual CH in the canal with apical transportation was higher than that in the canal without apical transportation (P<0.05). In conclusion, CH can be hardly removed completely. Canal with apical transportation will result in insufficient CH removal. EndoActivator, pre-curved ultrasonic file and F file are more effective in the curved root canal with apical transportation.

AIMTo study the mechanisms of protection of bcl-2 gene transfection against heat-stressed cardiomyocytes.

METHODSCardiomyocytes were isolated and cultured. bcl-2 was transfected into cardiomyocytes with Lipofectamine transfection methods. The cardiomyocytes were stressed by heat. The change of H+ -ATPase synthesis activity of cardiomyocytes mitochondria caused by bcl-2 transfection was measured by chemical radiation method. The changes of Caspase 3 activity of cardiomyocytes caused by bcl-2 transfection was measured by fluorometric analysis.

RESULTSbcl-2 transfection could increase the H+ -ATPase synthesis activity of cardiomyocytes mitochondria under heat stress at 41 degrees C and 43 degrees C and could decrease the Caspase 3 activity of cardiomyocytes under heat stress at 41 degrees C and 43 degrees C.

CONCLUSIONThe protection effect of bcl-2 transfection on heat-stressed cardiomyocytes may be associated with preserved H+ ATPase synthesis activity of cardiomyocytes mitochondria and the activity of Caspase 3 of cardiomyocytes.

Animals ; Caspase 3 ; metabolism ; Cells, Cultured ; Genes, bcl-2 ; Heat-Shock Response ; genetics ; Myocytes, Cardiac ; cytology ; metabolism ; Proton-Translocating ATPases ; metabolism ; Rats ; Transfection

Objective To evaluate ketamine-induced cerebral protection in mice with traumatic brain injury (TBI) by magnetic resonance imaging (MRI).Methods Thirty-two pathogen-free healthy male C57BL/6 mice,aged 8 weeks,weighing 26-30 g,were divided into 4 groups using a random number table:control group (group C,n=7),ketanine group (group K,n=7),TBI group (n=9) and TBI plus ketamine group (group TBI+K,n =9).TBI was produced with a pneumatically driven controlled cortical impact device.Ketamine 150 mg/kg was intraperitoneally injected at l h after operation in TBI+K and K groups,while the equal volume of normal saline was given instead in TBI and C groups.Open field test was conducted at 24 h,72 h and 7 days after operation.The animals in TBI and TBI+K groups were scanned by T1-weighted MRI at 6,24 and 72 h after operation,the animals in C and K groups were scanned by MRI at 24 h after operation,and the development of cerebral edema was observed.Results MRI scan showed no cerebral edema in C and K groups,and different degrees of cerebral edema were found in TBI and TBI+K groups.Compared with group C,the locomotor distance was significantly shortened at 24 and 72 h after operation in group TBI (P＜0.05).Compared with group TBI,the size of cerebral edema was significantly decreased,and the locomotor distance was prolonged at 24 and 72 h after operation in group TBI+K (P＜0.05 or 0.01).Conclusion MRI method further clarifies that ketamine can produce cerebral protection to some extent in mice with TBI.

Objective To investigate the radiosensitivity effects of poly ADP-ribose polymerase-1 (PARP-1) inhibitor 3-amion benzamide (3-AB) on the BRCA non-mutant and BRCA mutant breast cancer cells,and to explore the regulatory mechanism of PARP-1 and BRCA in radiation-induced DNA damage repair.Methods MDA-MB-436 cells and MDA-MB-231 cells were divided into four groups respectively as the control (CTRL),ionizing radiation alone (IR),3-AB alone (3-AB),and ionizing radiation combined with 3-AB(IR + 3-AB)group.γ-H2AX foci were detected by immunofluorescence assay.The radiosensitivity of breast cancer cells was evaluated by clonogenic survival assay.The percentage of apoptotic cells was assessed by flow cytometry.Results Compared with MDA-MB-231 cells,MDA-MB-436 cells had a higher radiosensitivity and produced more γ-H2AX foci(t =4.57,P ＜ 0.05),which was further increased by 3-AB.The DNA damage of MDA-MB-436 cells in the IR + 3-AB group was the most remarkable (t =3.26,P ＜ 0.05).Flow cytometry showed that the cells in the IR + 3-AB group had the highest rate of apoptosis (t=3.81,P ＜ 0.05),and the apoptosis rate of MDA-MB-436 cells was significantly higher than MDA-MB-231 cells (t =2.96,P ＜ 0.05).Conclusions The radiosensetivity of BRCA mutant cells MDA-MB-436 is significantly higher than that of non-BRCA mutant cells MDA-MB-231.Inhibition of PARP-1 can further increase the apoptosis and radiosensitivity of BRCA-mutant cells by further blocking the repair of DNA single-strand break induced by ionizing radiation.

Animals ; Dose-Response Relationship, Drug ; Male ; Mice ; Mice, Inbred Strains ; Nickel ; toxicity ; Oxidative Stress ; physiology ; Testis ; drug effects ; metabolism

Tooth or dentition missing compromises human health physically and psychiatrically. Although several prosthesis methods are used to restore tooth loss, these restorations are still non-biological methods. It is a dream for human being to regenerate a real tooth for hundreds years. There are two ways to regenerate the tooth. One is application of conventional tissue engineering techniques including seed cells and scaffold. The other is regeneration tooth using dental epithelium and dental mesenchymal cells based on the knowledge of tooth initiation and development. Marked progress has been achieved in these two ways, while there is still a long way to go. Recently a new concept has been proposed for regeneration of a biological tooth root based on tooth-related stem cells and tissue engineering technique. A biological tooth root has been regenerated in swine. It may be a valuable method for restoration of tooth loss before successful whole tooth regeneration. A latest research showed that a subpopulation in bone marrow cells can give rise to ameloblast-like cells when mixed with embryonic epithelium and reassociation with integrated mesenchyme, which may provide a new seed cell source for tooth regeneration.
Animals ; Bone Marrow Cells ; Epithelium ; Humans ; Mesenchymal Stromal Cells ; Regeneration ; Swine ; Tissue Engineering ; Tooth ; Tooth Root

OBJECTIVETo investigate the effect of HCMV infection on the expression of interleukin-8 (IL-8) and regulated on activation, normal T expressed and secreted (RANTES) so as to explore the possible mechanism of the immune-pathological changes caused by HCMV infection.

METHODSExpression of both IL-8 and RANTES mRNA was detected by RT-PCR. Secretion of IL-8 and RANTES protein was quantitated by enzyme linked immune-sororbant assay (ELISA).

RESULTSAfter HCMV infection, the expression of IL-8 in HEL cells was found to be increased gradually concomitant with the prolonged infection time at both the transcriptional level and the protein level. By the time of 72 h.p.i, as compared with mock-infected cells, IL-8 mRNA expression was increased up to 12-fold and IL-8 protein up to 9-fold in HCMV-infected cells. The expression of RANTES mRNA was occurred at 8 h.p.i, with a maximum at 24 h.p.i, since then it remained relatively high level. The expression of RANTES protein peaked at 24 h.p.i, then dropped sharply and was almost undetectable by the time at 72 h.p.i.

CONCLUSIONHCMV infection of HEL cells may induce the transcription of IL-8 gene as well as the production of IL-8 orotein. HCMV may down-regulate extra-cellular production of RANTES protein.

Cells, Cultured ; Chemokines ; genetics ; immunology ; Cytomegalovirus ; physiology ; Cytomegalovirus Infections ; genetics ; immunology ; virology ; Female ; Fibroblasts ; immunology ; virology ; Gene Expression ; Humans ; Pilot Projects ; Pregnancy