Objective To investigate the influence of two kinds of different cements (resin-reinforced glass ionomer and self-adhesive resin cement) to the pH of peri-implant crevicular fluid of cement retained implant restorations.Methods About 15 patients for cement retained implant restorations with a total of 30 dental implants were chosen for study.The implants were randomly divided into two groups,treatment group for the resin-reinforced glass ionomer,and control group for the self-adhesive resin cement,with 15 implants in each group.Another 15 patients were chosen to observe 1 healthy tooth and periodonatal tissue from each of them as blank group.The pH of peri-implant crevicular fluid and meliorate plaque index were measured respectively before the crowns were bonded,after 6 months and 12 months.The measured results during different periods were compared.Results Before the crowns were bonded,after 6 months and 12 months the pH of peri-implant crevicular fluid of treatment group were (7.19 ± 0.47),(7.30 ± 0.51),(7.38 ± 0.55),meliorate plaque index were (0.43 ± 0.35),(0.51 ± 0.36),(0.54 ± 0.45);the pH of peri-implant crevicular fluid of control group were (7.20 ± 0.55),(7.39 ± 0.48),(7.85 ± 0.53),meliorate plaque index were (0.42±0.41),(0.53 ± 0.45),(0.92 ± 0.59);the pH of gingival crevicular fluid of blank group were (7.21 ±0.52),(7.22 ±0.55),(7.29 ± 0.49),meliorate plaque index were (0.44 ±0.35),(0.48 ±0.25),(0.50 ± 0.46).Tweleve months after the crowns were bonded,pH of gingival crevicular fluid and meliorate plaque index in control group were significantly higher than that in treatment group and blank group.There was a positive correlativity between meliorate plaque index and pH of gingival crevicular fluid.Conclusion Resin-reinforced glass ionomer has less effect on the pH of peri-implant crevicular fluid.

OBJECTIVETo explore the killing effect of the mutant herpes simplex virus thymidine kinase (HSV-sr39tk) and its wild-type (HSV-tk) mediated by lentiviral vector on T lymphocytes in vitro and compare T cell survival rate after GCV or ACV treatment.

METHODSThe three-plasmid lentiviral vector system including packaging plasmid DeltaNRF, envelope plasmid VSV-G and vector plasmid (pTK151 + HSV-sr39tk or pTK151 + HSV-tk) were cotransfected into human embryonic kidney 293T cells using modified calcium phosphate precipitation methods. The packaged virus was harvested 72 h later. The survival of T cells expressing HSV-sr39tk or HSV-tk was measured by MTT assay after 4 day-culture against a gradient of GCV or ACV concentrations.

RESULTSThe three plasmids were effectively cotransfected and a high titre of lentivirus was obtained (2 x 10(6) IU/ml). 39tk(+) T cell survival rates declined promptly when the prodrug GCV/ACV concentrations increased from 0 micromol/L to 10 micromol/L. The T cell survival rates in GCV group declined from (96.04 +/- 3.23)% to (36.76 +/- 4.38)% while in ACV group from (97.31 +/- 4.61)% to (43.75 +/- 8.99)%. However, when GCV/ACV concentrations were more than 10 micromol/L, further decline of 39tk(+) T cell survival rates became unobvious. The growth rate of 39tk(+) T cell exposed to GCV or ACV was obviously lower than that in un-transfected T cells (P < 0.05). Tk(+) T cells were sensitive to GCV (P < 0.05), but not to ACV (P > 0.05). There was a significant difference in killing effects between 39tk(+) T cell + GCV group and tk(+) T cell + GCV group (P < 0.05).

CONCLUSIONThe lentiviral vectors containing HSV-sr39tk gene could infect T lymphocytes effectively and stably without affecting the proliferation of the transduced cell. In contrast to HSV-tk gene, T cells infected HSV-sr39tk were more sensitive not only to GCV but also to ACV.

Acyclovir ; pharmacology ; Animals ; Cell Survival ; drug effects ; Cells, Cultured ; Ganciclovir ; pharmacology ; Genetic Vectors ; Lentivirus ; genetics ; Mice ; Mice, Inbred C57BL ; Plasmids ; genetics ; T-Lymphocytes ; cytology ; drug effects ; Thymidine Kinase ; genetics ; Transfection

BACKGROUNDAscorbic acid (AA) represents one of the most important enzyme co-factors, antioxidants and neuromodulators and plays an important role in the cerebral system. Increasing evidence has suggested that AA could treat certain kinds of vertigo diseases such as Meniere's disease. To elucidate the neurochemical functions associated with AA in vertigo, the change of extracellular AA in the brain cortex following caloric vestibular stimulation (CVS) was evaluated.

METHODSAn on-line electrochemical detection was coupled with in vivo microdialysis to continuously monitor the change of extracellular AA in the primary somatosensory (SI) area of guinea pigs following a caloric vestibular stimulation. Sixteen guinea pigs were divided into three groups, i.e., experimental group with irrigation of the ear canal with ice water (0 degrees C) (n = 8), and two control groups, one with irrigation of the ear canal with warm water (38 degrees C) (n = 4) and the other with irrigation of the auricle with ice water (n = 4).

RESULTSIn the experimental group, the ice water irrigation of the left external ear canal induced a horizontal nystagmus towards the right side lasting about 45 seconds. No nystagmus was induced by warm water irrigation of the external ear canal or by ice water irrigation of the auricle. The extracellular AA concentration significantly increased following the ice water vestibular stimulation, reaching a maximum of (130 +/- 20)% (n = 8) of the basal dialysate level (2.61 +/- 0.92) micromol/L (n = 8), lasting at least for an hour. AA level did not change distinctly after the irrigation of the left external ear canal with warm water or the irrigation of the auricle with ice water.

CONCLUSIONSThe concentration of extracellular AA in the brain cortex of the SI area increased following the ice water vestibular stimulation. This demonstration may be useful for the investigation of the neurochemical processes associated with AA in the process of vertigo.

Animals ; Ascorbic Acid ; analysis ; Cerebral Cortex ; metabolism ; Electrochemistry ; methods ; Extracellular Space ; metabolism ; Guinea Pigs ; Ice ; Male ; Microdialysis ; methods ; Physical Stimulation ; methods ; Vestibule, Labyrinth ; physiopathology

Objective To explore the application of parents' presence induction program (PPIP) during anesthesia induction in children in the operation room and the nursing. Methods 86 children patients (1~11 years old) who would accept the operation were randomly divided into the observation and the control group (n=43 in each group). The observation group was accompanied by their parents when the children accepted the anesthesia induction under the instruction of nurses. But the patients were not present in the control group, and the patients were leaded into the operation room directly by the nurses to accept the anesthesia induction. Blood pressure and pulse changes before and after the anesthesia induction of these children patients were observed. The feeling of their heads of family before the operation and the satisfaction about PPIP were investigated. Results Blood pressure and pulses in the control group were significantly higher than those in the observation group before they were leaded into the operation room (P<0.01). 95% of heads of family hoped to accompany their children when they accepted the anesthesia induction, and 93.7% of heads of family wanted to take part in the PPIP. The satisfaction rate about PPIP was 95% after the completion of PPIP. Conclusions There are many profits about PPIP, such as mitigating the fear and anxiety of children and relaxing the unstable feeling of their heads of families before the anesthesia, and reducing the quantity of pre-anesthetic medications. These are helpful to the anesthesia induction and the cooperation of doctor patients. It is a best anesthesia induction mode worthy to be extended in the operation room.

OBJECTIVETo establish strategies for preventing graft versus host disease (GVHD) and reducing treatment associated morbidity while preserving graft versus leukemia (GVL) effect in nonmyeloablative allogeneic bone marrow transplantation (allo-BMT), with or without donor lymphocyte infusion (DLI) after BMT.

METHODS3 x 10(7) bone marrow cells mixed with 1 x 10(7) spleen cells from the same BALB/c mouse were transplanted into the nonablative irradiated inbred 615 mouse which received a single subcutaneous injection of 1 x 10(6) L615 leukemia cells three days before. The experiments were designed as follows (ten mice in each group): myeloablative BMT control group (group A), nonmyeloablative conditioning without BMT group (group B), nonmyeloablative BMT group (group C), and nonmyeloablative BMT + DLI group (group D). GVL effects were assessed by survival time, white blood cell count and L615 cells in peripheral blood and histologic changes. GVHD was assessed by signs of weight loss, ruffled fur, diarrhea and histologic changes of skin, liver and small intestines. Chimerism was detected by cytogenetic analysis and PCR technique.

RESULTSThe survival time of group A, B, C and D was (20.3 +/- 13.4), (15.9 +/- 1.1), (21.6 +/- 1.7) and (37.8 +/- 2.0) days, respectively, being no significant difference between group A and group C (P > 0.05). The survival time of group C was longer than that of group B (P < 0.01). And among group B, C and D, group D had the longest survival time (P < 0.01). GVHD signs and histologic changes were observed in 60% of control group mice at + 14 day, but none of group C and group D. 40% of mice in group A died of treatment associated morbidity within two weeks, but none in group C and group D. Allogeneic chimerism was kept in group A, but excluded gradually in group C.

CONCLUSIONGVL effect seems preserved in nonmyeloablative BMT mice, but weaker than that in myeloablative BMT mice. GVL effect seems to be enhanced by DLI after nonmyeloablative BMT. GVHD and transplantation associated morbidity seems to be reduced in nonmyeloablative BMT.

Animals ; Bone Marrow Transplantation ; immunology ; methods ; Combined Modality Therapy ; Female ; Graft vs Host Disease ; prevention & control ; Graft vs Leukemia Effect ; Leukemia, Experimental ; therapy ; Leukemia, Lymphoid ; therapy ; Lymphocyte Transfusion ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Transplantation Conditioning ; methods ; Transplantation, Heterologous

OBJECTIVETo construct a three plasmids lentiviral vector containing canine coagulation factor IX (cFIX) gene with ubiquinone promoter (PUB) and observe the expression of cFIX gene.

METHODSLentivirus was generated by transient three-plasmid transfection, namely, the VSV-G envelope expression cassette, the delta NRF packaging plasmid and the PTK 164 plasmid. Viral particles were used to infect the target cell, third passage mesenchymal stem cells (MSCs) and 293T cell respectively at MOI 3: 1. The cFIX activity was detected in cultured cells with one-stage clotting assay.

RESULTSThe MSCs were obtained in vitro. The lentivirus infected MSCs and 293T cells all expressed the active factor IX with the activity of (26.30 +/- 2.10)% and (19.70 +/- 1.53)%, respectively, which are significantly higher than that of control (1.00 +/- 0.05)%.

CONCLUSIONSThe lentiviral vector of three plasmids with ubiquinone promoter (PUB) was constructed and can transfect the MSCs and 293T cells.

Animals ; Bone Marrow Cells ; metabolism ; Cells, Cultured ; Dogs ; Factor IX ; genetics ; metabolism ; Genetic Vectors ; Hemophilia B ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; Plasmids ; genetics ; Transfection

implementation process using the process of clinical pathway in intraspinal tumor resectionas a case study . Methods Take Delphi method of experts for three rounds , and finally to determine the evaluation system of clinical pathway implementation process .Results The recovery rate of the questionnaire on expert consultationwas 100%.The third round of expert investigation showed that the overall degree of authority was about 0.84, the average scores of the importance of each index from experts were all from 4 (important) to 5 (very important ), and the coefficient of variation was fluctuation in the range of 0.112 ~0.285. Conclusions Experts who take part in the investigation are full of enthusiasm , the viewpoints of the importance of each index from experts are relatively concentrated , the coordination degree is higher , and the results of the survey have higher reliability.

BACKGROUND: Mesenchymal stem cells are derived from a variety of tissues, such as bone marrow, pulp, placenta, umbilical cord and adipose tissue. Mesenchymal stem cells from deciduous pulp have strong stemness and biological activity, no rejection, and strong immunoregulation, which are one of excellent cell sources for biotherapy. It is easy and suitable for large-scale production of mesenchymal stem cells from deciduous pulp, thereby laying a good foundation for the industrialization of dental pulp stem cells. OBJECTIVE: To investigate the effect of primary tooth root resorption on the isolation and expansion of dental pulp stem cells, in order to further determine the proper period for tooth extraction for pulp stem cell isolation. METHODS: Totally 173 primary teeth from 173 pupils aged 7-9 years were extracted for the isolation and expansion of dental pulp stem cells. Before tooth extraction, we took X-ray periapical film or orthopantomography of the primary teeth, in accordance with the World Health Organization (WHO) professional inspection standard. Root resorption in primary teeth could be divided into five kinds: root resorption 1/3, root resorption 1/2, root resorption 2/3, complete root resorption, and natural loss of primary teeth. Collected teeth after tooth extraction were placed into a medium within 7 seconds, and stored at in a refrigerator of 2-4 ℃. Then, the teeth were sent to the Oral Stem Cell Bank in Beijing within 24 hours by a professional cold-chain logistics for the isolation, expansion and preservation of dental pulp stem cells. Statistical analysis of the test results was performed. RESULTS AND CONCLUSION: For 32 primary teeth with root resorption 1/3, dental pulp stem cells were successfully extracted from 30 teeth, with a success rate of 94%, and ectopic eruption of permanent teeth was found in 12 cases, with an average eruption time of (2.19±0.18) months. For 35 primary teeth with root resorption 1/2, dental pulp stem cells were successfully extracted from 32 teeth, with a success rate of 92%, and ectopic eruption of permanent teeth was found in 11 cases, with an average eruption time of (1.89±0.13) months. For 59 primary teeth with root resorption 2/3, dental pulp stem cells were successfully extracted from 54 teeth, with a success rate of 92%, and ectopic eruption of permanent teeth was found in 8 cases, with an average eruption time of (1.42±0.12) months. For 37 primary teeth with complete root resorption (the bottom of the pulp was intact), dental pulp stem cells were successfully extracted from 34 teeth, with a success rate of 92%, and ectopic eruption of permanent teeth was found in 2 cases, with an average eruption time of (1.03±0.15) months. For 10 naturally exfoliated primary teeth, dental pulp stem cells were not extracted, and ectopic eruption of permanent teeth was found in 4 cases, with an average eruption time of (0.65±0.23) months. To conclude, the primary teeth naturally exfoliated have no dental pulp with no stem cells; the success rate of extraction is relatively high in primary teeth that have mobility I-II, root resorption 2/3 or complete root resorption but with the complete bottom of the pulp. Moreover, it has no effect on permanent tooth eruption, and it is the best time for collection of primary teeth.

Humans ; Hydrogen-Ion Concentration ; Malassezia ; growth & development ; physiology ; Staphylococcus epidermidis ; growth & development ; physiology

Objective To investigate the epidemioiogical pattern of Borna disease virus (BDV) among different canine breeds in Ili, China, and to analyze its potential phylogeny. Methods BDV p24 RNA fragments were detected from peripheral blood mononuclear cells (PBMCs) of canine by modified nested RT-PCR (nRT-PCR). Possible false positives were excluded by determination of both BDV p40 RNA fragments and PMD19 plasmid standards. Analysis were performed on genetic sequence, homologous comparison, amino acid sequence and phylogeny after p24 positive products were validated. Results BDV p24 RNA fragments were found only in Kazakh Tobet (a shepherd dog) in 8 breeds of 150 cases and their overall positive rate was 11.0% (10/91). Compared with the strain of He/80 from horse and that of S6 from sheep in Germany, the homologous similarities of Kazakh Tobet was 99.2% and 95.7%, and that of amino acid as 100% and 89.3%, respectively. The kinship of Kazakh Tobet was close to He/80 and next to S6. Conclusion There was potential natural BDV infection in Kazakh Tobet in Ili, and its endemic strain was concerned with He/ 80 infecting Ili horse and S6 of German Merino sheep introduced into the region from Germany.