Effect of autologous platelet-rich plasma in combination with bovine porous bone mineral and bio-guide membrane on bone regeneration in mandible bicortical bony defects.

OBJECTIVE: Growth factors contained in platelet-rich plasma (PRP) can induce osteoblast differentiation in certain studies, whereas in others, osteogenesis of PRP on mandible bone defects has not been proved clinically. The aim of the study was to investigate the effect of autologous PRP on the osteogenic potential of combining bovine porous bone mineral (BPBM) and bio-guide membrane (BGM) in promoting mandible bicortical bony defects in rabbits. METHODS: One circular mandible bicortical bony defects were created in each of 54 rabbits, which were divided into 3 groups: group 1: 18 of the defects were left unfilled as a negative control; group 2: 18 of the defects were grafted with autologous PRP and BPBM/BGM; group 3: 18 of the defects were grafted with BPBM/BGM without PRP. Animals were killed at 4, 8, and 12 weeks after operation. Harvested tissue and specimens were evaluated histologically and radiographically, and metabolized observation was performed. Histological parameters associated with osteoblast activities, bone trabecula, neovascularization, newly formed mineralized bone, rudimental grafts and connective tissue formation were measured. Densities of the bones at 4, 8, and 12 weeks were studied by radiographic. The bone defect closure ratio was measured at 12 weeks. The bone metabolized parameter alkaline phosphatase was also measured and compared between 4, 8, and 12 weeks. RESULTS: The platelet concentration of PRP is 4.19- to 4.43-fold to that of the whole blood. Histological analysis showed new bone formation at all therapeutic sites including BPBM/BGM grafts with or without PRP. A statistically significant difference in new bone formation between group PRP/BPBM/BGM and group BPBM/BGM was observed. Untreated defects of group control showed the less bone regeneration. There was significant difference of bone density between group PRP/BPBM/BGM and control, and group BPBM/BGM and control, at 4, 8, and 12 weeks postoperative. There were more bone defects filling, and the grafts were absorbed at 12 weeks of group PRP/BPBM/BGM compared with group BPBM/BGM. Defects treated with PRP/BPBM/BGM demonstrated significantly increased activity of osteoblasts, enhanced amount of mitochondria and rough endoplasmic reticulum in osteoblasts, and increased concentration of alkaline phosphatase at 4, 8, and 12 weeks compared with those treated with BPBM/BGM and control group. Complete closure ratio of bone defects treated with PRP/BPBM/BGM (50%) was significantly increased compared with that treated with BPBM/BGM (16.6%). CONCLUSIONS: The study suggested that PRP combination of BPBM and BGM had significant therapeutic effects on mandible bicortical bony defects of rabbits. The effects are associated with the high concentration of platelet in PRP and the porous configuration of BPBM. Although we cannot reveal the detailed statistical relationship of PRP on promoting BPBM/GBM osteoinductive effects, PRP demonstrated superior results of bone regeneration.

Distortion of terahertz signals due to imperfect synchronization with chirped probe pulses.

Terahertz (THz) signals measured by means of the spectral-encoding technique with different temporal discrepancies between probe pulses and THz signals are investigated. It is found that imperfect synchronization between the chirped probe and THz pulses induce a distortion and this distortion affects significantly the retrieved THz spectrum if the temporal discrepancy is large. The distortion becomes more prominent if the probe pulse length is less than the optimal chirped probe pulse duration. A simple approach is proposed to realize the synchronization and minimize the distortion. THz signals from a high-voltage-biased air plasma filament are measured with this approach and distortion similar to the simulation results is observed.

Therapeutic effects of diode laser on vascular epulis in esthetic area.

Vascular epulis is a rare clinical disease. In our study, a case of vascular epulis in the cosmetic area was treated by diode laser, without recurrence and obvious inflammation in the surgical site 5 years after surgery. This case report indicates that the excision of vascular epulis in the cosmetic area of the anterior teeth by diode laser could be an alternatively safe and complementary approach in lieu of conventional surgery.

Acoustic Properties of the Otolith of the Large Yellow Croaker Larimichthys crocea (Perciformes: Sciaenidae).

The inner ears of fish contain three pairs of otoliths-lapilli, asterisci and sagittae-which play important roles in hearing and balance. However, acoustic properties and dynamic responses of fish otoliths are poorly understood. The large yellow croaker (Larimichthys crocea), like many species in the family Sciaenidae, is extremely sensitive to sound. The present study used L. crocea sagittae as the research subject and examined the variation in shear stress on sagittae under different acoustic stimuli. For the first time, the sound speed of the sagitta was measured using ultrasonic pulse-echo techniques, and the acoustic impedance and natural frequency of the sagitta were calculated. Larimichthys crocea adults (20-22 cm standard length, n = 10) had a sagitta density of 2781.5 ± 28.06 kg/m3, sound speed of 4828-6000 m/s and acoustic impedance range of 13.4-16.7 MPa·s/m, approximately 9-11 times that of seawater (1.48 MPa·s/m). The natural frequency of the sagitta was 76.4-95.5 kHz. The shape and structural details of sagittae were reconstructed by 3D scanner and the shear stress responses of sagittae under different acoustic stimulus were investigated based on a finite element model. The simulation results showed that the shear stress responses tended to increase and then decrease in the range of sciaenid hearing frequency from 200 to 1300 Hz, peaking at 800 Hz. The shear stress responses varied with the direction of acoustic stimulus and peaked when the incident direction was perpendicular to the inner surface of the otolith. These results provide important parameters that may be used to protect L. crocea from possible underwater noise damage, particularly during their spawning aggregations and over-wintering aggregations.

Facile formation and redox of benzoxazole-2-thiolate-bridged dinuclear Pt(II/III) complexes.

Reaction of [Pt(L)(µ-Cl)](2) (L = ppy (2-phenylpyridine) or bzq (benzo[h]quinoline)) with 2-mercaptobenzoxazole (NOSH) and NaOAc in THF at r.t. yields the dinuclear Pt(II) d(8)-d(8) complexes [Pt(2)L(2)(µ-NOS-κN,S)(2)] (L = ppy, 1; L = bzq, 2) and the Pt(III) d(7)-d(7) complexes [Pt(2)(ppy)(2)(µ-NOS-κN,S)(2)(NOS-κS)(2)] (L = ppy, 3; L = bzq, 4) in one pot. The C,N-cyclometalated ligand is chelating whereas the N,S-donating benzoxazole-2-thiolates doubly bridge the two metal centers. The Pt···Pt separations of 3.0204(3) and 2.9726(8) Å in 1 and 2 contract to 2.685(1) Å in 3 and 2.6923(3) Å in 4, respectively, when two S-bound thiolate ligands coordinate trans- to the Pt···Pt axis. However, cyclometalation is preserved and there is minimum perturbation of the bridging ligands. Complexes 3 and 4 can be also obtained by oxidative addition of the thiolate ligand. In the presence of NaBH(4), 3 and 4 can be reduced to 1 and 2, respectively. At r.t., 1 and 2 exhibit intense orange-red luminescence at 625 nm and 631 nm, respectively. The electrochemical properties of 1-4 have been also discussed.

Intrinsically fluorescent carbon dots with tunable emission derived from hydrothermal treatment of glucose in the presence of monopotassium phosphate.

A facile method is developed to synthesize intrinsically fluorescent carbon dots by hydrothermal treatment of glucose in the presence of monopotassium phosphate. The fluorescence emission of the carbon dots thus produced is tunable by simply adjusting the concentration of monopotassium phosphate.

[Multiple supernumerary teeth: a case report].

Supernumerary teeth is one of the dysplasia that the number of the teeth are more than physical number. Most cases of reports were with 1-2 supernumerary teeth and rare cases were with more than 3 supernumerary teeth. A 17-year old female patient of 7 impacted supernumerary teeth were found because of toothache of premolar caused by impacted supernumerary teeth and were treated by extraction of impacted supernumerary teeth.

[Maxillary second molar with two buccal and two lingual root canals: a case report].

Incidence rate of 4 root canals in maxillary second molar is very low and most molars have only two mesiobuccal root canals. The emergence of 4 root canals in maxillary second molar with two lingual root canals is especially rare. A case of 4 root canals maxillary second molar with two lingual root canals was successfully treated and reported in this article.

Asterosaponin 1, a cytostatic compound from the starfish Culcita novaeguineae, functions by inducing apoptosis in human glioblastoma U87MG cells.

Malignant glioblastoma is one of the most common malignant tumors in the neurological system. Asterosaponin 1, a new cytostatic agent from the starfish Culcita novaeguineae appear to exhibit various biological activities, including antitumor effect, but the function and mechanism of this new agent on glioblastoma cells has not previously been determined. In the present study, we investigated the proliferation change of human glioblastoma U87MG cells exposed to different concentrations (2.5-20.0 microg/ml) of asterosaponin 1 for a certain time. The results showed that asterosaponin 1 significantly suppressed U87MG cell proliferation in a time- and dose-dependent manner (IC50 =4.3 microg/ml). Flow cytometric analysis of DNA in U87MG cells showed that asterosaponin 1 induces the prominent appearance of a sub-G1 peak in the cell cycle suggestive of apoptosis identical with the result of annexin V/PI assay. Furthermore, U87MG cells treatment with asterosaponin 1 resulted in nuclear condensation with apoptotic bodies observed by both fluorescence and electron microscopy. Agarose gel electrophoresis of DNA from asterosaponin 1-treated cells revealed a typical "ladder" consistent with apoptotic DNA fragmentation. Western-blot staining showed asterosaponin 1 decreased the expression of Bcl-2 protein and increased the expression of Bax protein. The novel findings suggest that the cytostatic actions of asterosaponin 1 toward U87MG cells result from the induction of cell apoptosis. Overall, our data demonstrate that asterosaponin 1 is fully equipped for an efficient apoptotic killing of glioblastoma cells and suggest that this mechanism may play a critical role in anti-tumor chemotherapy.

[The effects of rBCG expressing Der p2 in the form of lipoprotein on murine immune response].

AIM: To investigate the effects of rBCG vaccination containing foreign antigen Der p2 in the form of lipoprotein on murine immune response. METHODS: 6 to 8 weeks old and newborn BALB/c mice were vaccined intraperitoneally with 10(6) CFU rBCG or BCG. At the same time, the control group was injected with saline. Six weeks later, all animals were injected with Der p2 (20 microg). After two weeks later, the concentrations of IL-4 and IFN-gamma in the serum and splenocyte culture supernatant (STLCS) were determined by ELISA, and Th subgroups were determined by double fluorescent staining and flow cytometry. RESULTS: After vaccination, the serum and STLCS from both rBCG-immunized and BCG-immunized group of adult and newborn BALB/c mice had significantly higher level of IFN-gamma and lower level of IL-4 than those from control groups. Besides, there was the larger percentage of CD4 (+) IFN-gamma (+) cells in spleen from rBCG-vaccined and BCG-vaccined mice than that from control group. However, the percentage of CD4 (+) IL-4 (+) cells in spleen cells from rBCG-vaccined and BCG-vaccined group was lower than that from control group. Moreover, the level of IFN-gamma in STLCS from rBCG-immunized was significantly higher, compared with that from BCG-immunized mice. At the same time, the percentage of CD4 (+) IFN-gamma (+) cells in spleen from rBCG-vaccined mice was larger than that from BCG-vaccined group. CONCLUSION: Both rBCG and BCG could stimulate Th1 predominant immune response, when injected intraperitoneally into adult or newborn BALB/c mice, The Der p2 expressed on the cell wall of BCG can work as the component of BCG and be recognized by the immune system of mice, therefore stimulates Der p2-specific Th1 predominant immune response. These data indicate that recombinant BCG-expressing antigens can be used as the antigen-specific vaccines against allergic diseases by regulating the balance of Th1/Th2.

[Morphological evidence on participating in adhesive function of human umbilical vein endothelial cells by CD226].

AIM: To provide the morphological evidence on participating in the adhesive function of human umbilical vein endothelial cells by CD226. METHODS: Human umbilical vein endothelial cells(HUVECs) were cultured and the effect of CD226 on the adhesion between activated HUVEC and activated PBMC were observed by Leica inverted microscope and scanning electron microscope. RESULTS: Fusion protein CD226/Ig could block the adhesion between activated HUVECs and activated PBMC markedly. CONCLUSION: CD226 was a novel inducible adhesion molecule expressed on activated endothelial cells, which may possess important physiological and pathological function.

[Cloning, prokaryotic expression of LAIR and identification of their immunological reactivities].

AIM: To clone and express the leukocyte-associated immunoglobulin-like receptor (LAIR) and to identify the immune reactivity of LAIR-2 to anti-LAIR-1 specific monoclonal antibodies(mAb). METHODS: Genes encoding LAIR-1 and LAIR-2 were cloned by RT-PCR from human peripheral blood mononuclear cells(PBMC) and two leukemia cell lines Jurkat and HL-60. The extracellular region gene of LAIR1 and LAIR-2 were inserted into vector pGEX-4T-3 expressing GST fusion protein, expressed on IPTG induction and purified through glutathione-sepharose 4B column. The immunological reactivity of expressed LAIR-2 to anti-LAIR-1 mAb was identified by indirect ELISA. RESULTS: LAIR-1 and LAIR-2 cDNAs had been cloned and expressed. Five new LAIR-1 cDNA isoforms were cloned. Among them, two isoforms from HL-60 included LAIR-1 open reading frames (ORF) and three isoforms from Jurkat were LAIR-1 cDNA segments. The LAIR-1 and LAIR-2 showed different immunological reactivities. CONCLUSION: The transcription, processing after transcription and expression of LAIRs may be related to disparities in individuals and disease status. The difference in immunological reactivity may be involved in their structure.

[Gene cloning and prokaryotic expression of the PID domain of human DOC-2 molecule].

AIM: To clone the PID domain of human DOC-2 (nDOC-2) and express it in E.coli DH5alpha. METHODS: The cDNA fragment encoding the PID domain of nDOC-2 was amplified by RT-PCR from normal human ovarian tissue and cloned to pUC19. The DNA fragment from the pUC19-nDOC-2 digested with BamH I and EcoR I was ligated to the BamH I/EcoR I digested prokaryotic expression vector pGEX-4T-1.The expression of fusion protein was induced with IPTG and the expressed product was identified by SDS-PAGE. RESULTS: (1)The sequencing and endonucleases digestion analysis showed that the fragment of nDOC-2 gene was insert into vectors pUC19 and pGEX-4T-1;(2)SDS-PAGE showed the nDOC-2 gene had been expressed in E.coli DH5alpha. CONCLUSION: The PID domain of nDOC-2 was expressed successfully in prokaryote, which makes preparation for further researching the function of DOC-2 and preparing antibodies to DOC-2 protein.

[Eukaryotic expression and application of human CD226 Ig fusion protein containing 3C protease-restricted site].

AIM: To construct, express and characterize the eukaryotic expression vector of encoding human CD226 (PTA1) extracellular region Ig fusion protein gene containing 3C protease-restricted site. METHODS: The gene fragment encoding extracellular region of human CD226 was cloned into the eukaryotic expression vector p-3C-Ig containing 3C protease-restricted site and human Ig Fc fragment gene. After sequencing, the vector was transfected into COS7 cells, and the expressed molecule was purified by affinity chromatography. Finally, the product was characterized by immunofluorescent staining and 3C protease digestion. RESULTS: After expression and purification, the Ig fusion protein could bind effectively to the CD226 ligand expressed on ECV304 cells. The Fc fragment could also be cut off by 3C protease, so that the CD226 extracellular fragment was obtained. CONCLUSION: The extracellular region of human CD226 Ig fusion protein with 3C protease-restricted site is expressed successfully in COS-7 cells, which lays the foundation for the structural and functional study of this molecule.

[Effects and holding time of hyperbaric oxygen on human severe periodontitis].

OBJECTIVE: This paper studied the therapeutic effects and holding time of hyperbaric oxygen (HBO) on human severe periodontitis. METHODS: 30 cases with periodontitis were selected and randomly divided into 2 groups, i.e. the HBO group and control group. For HBO group, they were exposed to a pressure of 0.25 MPa. For control group, they were rinsed with gargle. Gingival indices (GI), sulcus bleeding indices (SBI), plaque index (PLI), probing depth (PD), attachment loss (AL) and gingival crevicular fluid (GCF) were measured during both the first and last clinical visits, and 1 year after HBO therapy. The gingival blood flow (GBF) were measured by Laser Doppler Flowmeter. RESULTS: HBO can decrease GI of patients with periodontitis by 1.1 decrease SBI by 1.2, lower PD and AL by 0.7 mm, decrease the volume of GCF by 2.0, and significant differences could be seen in the above indices between pre and post HBO therapy. The GBF had a 1.8 folds increase after HBO exposure. GI and SBI one year after HBO therapy were larger than that of the time after HBO therapy. There were no significant differences in the PLI, PD, AL, GCF, GBF between post HBO therapy and 1 year after HBO therapy. CONCLUSION: HBO had good therapeutic effects on human severe periodontitis, the effects can keep more than 1 year.