Efeito da associação de tratamentos com fosfopeptídeo de caseína-fosfato de cálcio amorfo e trimetafosfato de sódio sobre a desmineralização e remineralização do esmalte: estudos in vitro / Effect of the combination of treatments with casein phosphopeptide-amorphous calcium phosphate and sodium trimetaphosphate on desmineralization and enamel remineralization: in vitro studies

A proposição geral do presente estudo foi avaliar in vitro a associação de tratamentos com dentifrícios fluoretados e suplementados com trimetafosfato de sódio (TMP) e fosfopeptídeo de caseína-fosfato de cálcio amorfo (CPP-ACP) (MI Paste Plus®) em promover a remineralização e reduzir a desmineralização, respectivamente, do esmalte dentário. Blocos de esmalte bovinos (12/grupo) foram selecionados através da dureza de superfície inical (SH) e divididos em 5 grupos experimentais: 1) Dentifrício sem F (Placebo); 2) Dentifrício com 1100 ppm F (1100F), 3) MI Paste Plus®, 4) Dentifrício com 1100 ppm F associado a MI Paste Plus® (1100F-MI Paste Plus®) e 5) Dentifrício com 1100 ppm F + 3%TMP associado a MI Paste Plus® (1100F-TMPMI Paste Plus®). Para o Artigo 1 de Remineralização (RE>DES), blocos de esmalte bovino foram selecionados pela dureza de superfície pós-lesão de cárie artificial (SH1) e submetidos a 6 ciclagens de pH por 6 dias. Após as ciclagens de pH, foram determinadas dureza de superfície final (SH2), para o cálculo da porcentagem de recuperação de dureza de superfície (%SHR), perda integrada de dureza de subsuperfície (ΔKHN), análise do perfil e profundidade das lesões de subsuperfície através da microscopia de luz polarizada (PLM), microsopia confocal de varredura à laser (MCVL), microscopia eletrônica de varredura (MEV), espectroscopia de energia dispersiva (EDS), concentração de fluoreto (F), cálcio (Ca) e fósforo (P) no esmalte. Os dados foram submetidos à ANOVA (1-critério), seguido pelo teste StudentNewman-Keuls (p < 0,001). Os grupos 1100F e 1100F-TMP-MI Paste Plus® apresentaram valores semelhantes de %SHR (p = 0,150). A menor profundidade de lesão (ΔKHN e PLM) foi observada para o grupo 1100F-TMP-MI Paste Plus® quando comparado aos demais (p < 0,001). O grupo 1100F-TMP-MI Paste Plus® apresentou superfície mais uniforme e íntegra em relação aos demais tratamentos (MCVL e MEV). A concentração de F foi similar entre os grupos 1100F, 1100F-MI Paste Plus® e 1100F-TMP-MI Paste Plus® (p > 0,001). O tratamento com 1100F-TMP-MI Paste Plus® promoveu um aumento na concentração de Ca no esmalte em ⁓ 51% e ⁓ 21% respectivamente, quando comparado aos grupos 1100F e MI Paste Plus® (p < 0,001). Valores semelhantes de P no esmalte foram observados nos grupos MI Paste Plus®, 1100F-MI Paste Plus® (p > 0,001), exceto o grupo 1100F-TMP-MI Paste Plus®, que apresentou alta concentração (p < 0,001). Para o Artigo 2 de Desmineralização (DES>RE), blocos de esmalte bovino foram selecionados pela dureza de superfície inicial (SHi) e a seguir submetidos a 5 ciclagens de pH por 7 dias. Após determinou-se dureza de superfície final (SHf), porcentagem de perda de dureza de superfície (%SH), perda integrada de dureza de subsuperfície (ΔKHN), análise do perfil e profundidade das lesões de subsuperfície através da microscopia de luz polarizada (PLM), microsopia confocal de varredura à laser (MCVL), microscopia eletrônica de varredura (MEV), espectroscopia de energia dispersiva (EDS), concentração de fluoreto (F), cálcio (Ca) e fósforo (P) no esmalte. Os dados foram submetidos à ANOVA (1-critério), seguido pelo teste Student-Newman-Keuls (p < 0,001). Para a %SHR, o grupo Placebo apresentou os menores valores (p > 0,001). O grupo 1100F-TMP-MI Paste Plus® remineralizou a superfície do esmalte em ~ 38% em relação ao MI Paste Plus® (p < 0,001). A menor profundidade da lesão (ΔKHN) foi observada para o grupo 1100F-TMP-MI Paste Plus® quando comparado aos demais (p < 0,001), sendo inferior em 32% quando comparado ao grupo 1100F. A concentração de F, Ca e P foi maior para o grupo 1100F-TMP-MI Paste Plus® (p > 0,001). Diante dos resultados parciais obtidos, é possível concluir que a associação de tratamentos com dentifrícios fluoretados e suplementados com trimetafosfato de sódio (TMP) e fosfopeptídeo de caseína-fosfato de cálcio amorfo (CPP-ACP) (MI Paste Plus®) (1100F-TMPMI Paste Plus®) promoveu um efeito adicional significativo no processo de desremineralização, podendo ser uma alternativa de tratamento para pacientes em risco e atividade de cárie(AU)

The general purpose of this study was to evaluate in vitro the association of treatments with fluoridated toothpastes and supplemented with sodium trimetaphosphate (TMP) and casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) (MI Paste Plus®) in promoting remineralization and reduce demineralization, respectively, of tooth enamel. Bovine enamel blocks (12/group) were selected through the initial surface hardness (SH) and divided into 5 experimental groups: 1) Toothpaste without F (Placebo); 2) Toothpaste with 1100 ppm F (1100F), 3) MI Paste Plus®, 4) Toothpaste with 1100 ppm F associated with MI Paste Plus® (1100F-MI Paste Plus®) and 5) Toothpaste with 1100 ppm F + 3% TMP associated with MI Paste Plus® (1100F-TMP-MI Paste Plus®). For Remineralization Manuscript 1 (RE>DES), blocks of bovine enamel were selected for the surface hardness after artificial caries lesion (SH1) and subjected to 6 pH cycles for 6 days. After pH cycling, final surface hardness (SH2) was determined to calculate the percentage of surface hardness recovery (%SHR), integrated loss of subsurface hardness (ΔKHN), profile analysis and depth of the lesions of subsurface through polarized light microscopy (PLM), confocal laser scanning microscope (MCVL), scanning electron microscopy (SEM), dispersive energy spectroscopy (EDS), fluoride (F), calcium (Ca) and phosphorus (P) concentration in the enamel. The data were submitted to ANOVA (1-criterion), followed by the Student-Newman-Keuls test (p<0.001). 1100F and 1100F-TMP-MI Paste Plus® groups showed similar values of %SHR (p = 0.150). The lowest depth of lesion (ΔKHN and PLM) was observed for the 1100F-TMP-MI Paste Plus® group when compared to the others (p<0.001). The 1100F-TMP-MI Paste Plus® group showed a more uniform and complete surface in relation to the other treatments (MCVL and SEM). The F concentration was similar between the 1100F, 1100F-MI Paste Plus® and 1100F-TMP-MI Paste Plus® groups (p>0.001). The treatment with 1100F-TMP-MI Paste Plus® promoted an increase in the concentration of Ca in the enamel by ⁓ 51% and ⁓ 21% respectively, when compared to the 1100F and MI Paste Plus® groups (p<0.001). Similar values of P in the enamel were observed in the MI Paste Plus®, 1100F-MI Paste Plus® groups (p>0.001), except for the 1100F-TMP-MI Paste Plus® group, which presented high concentration (p<0.001). For demineralization Manuscript 2 (DES> RE), bovine of enamel blocks were selected for their initial surface hardness (SHi) and then subjected to 5 pH cycles for 7 days. After final surface hardness (SHf), percentage of loss of surface hardness (%SH), integrated loss of subsurface hardness (ΔKHN), analysis of the profile and depth of subsurface lesions through polarized light microscopy ( PLM), confocal laser scanning microscopy (MCVL), scanning electron microscopy (SEM), dispersive energy spectroscopy (EDS), fluoride (F), calcium (Ca) and phosphorus (P) concentration in the enamel. The data were submitted to ANOVA (1-criterion), followed by the Student-Newman-Keuls test (p<0.001). For %SHR, the Placebo group had the lowest values (p>0.001). The 1100F-TMP-MI Paste Plus® group remineralized the enamel surface by ~ 38% compared to MI Paste Plus® (p<0.001). The lowest depth of the lesion (ΔKHN) was observed for the 1100F-TMP-MI Paste Plus® group when compared to the others (p<0.001), being 32% lower when compared to the 1100F group. The F, Ca and P concentration was higher for the 1100F-TMP-MI Paste Plus® group (p>0.001). In view of the partial results obtained, it is possible to conclude that the combination of treatments with fluoridated toothpastes and supplemented with sodium trimetaphosphate (TMP) and amorphous calcium phosphate casein-phosphate (CPP-ACP) (MI Paste Plus®) (1100F-TMP -MI Paste Plus®) promoted a significant additional effect in the de-remineralization process, and could be an alternative treatment for patients at risk and caries activity(AU)

Effect of rgs4 overexpression on related protein expression of mglur5-mediated signaling pathway in methamphetamine-dependent rat striatum / 中国药理学通报

Aim To investigate the effect of the regulator of G-protein signaling 4(RGS4) overexpression in rat striatum on the related protein expression of metabolic glutamate receptor 5(mGluR5) signaling pathway and the conditioned place preference(CPP) behavior in rats, by establishing the METH-dependent CPP model. Methods Rats were divided into five groups: Normal, normal saline(NS), METH, Ad5-RGS4-EGFP and Ad5-EGFP group. Normal group was without any administration, while the striatum of the other groups were respectively stereotactic injected with phosphate buffer methamphetamine (METH)-depardent soline (PBS), PBS, overexpressed adenovirus vector Ad5-RGS4-EGFP and negative control adenovirus vector Ad5-EGFP. The CPP behavior of rats in each group was analyzed. The expression of RGS4, mGluR5, Gαq, PLC1 was measured in rat striatum tissue by Western blot. Results The difference of CPP in Ad5-RGS4-EGFP group decreased compared with that in METH group and Ad5-EGFP group(P 0.05). PLCβ1 expression changed with no significant difference. Conclusions RGS4 overexpression in striatum is able to alleviate the CPP behavior in METH-dependent rats, and its mechanism may be associated with the overexpression of RGS4 in METH-dependent rat striatum, which could down-regulate the mGluR5-mediated Gαq and PLCβ1 signaling pathway.

A prospective, double blind randomised controlled trial on efficacy of laparoscopic uterosacral nerve ablation as an alternative of pharmacotherapy in treatment of chronic pelvic pain

Background: Chronic Pelvic Pain (CPP) is one of the commonest symptomatology in gynaecologist’s outpatient clinics. CPP has a profound impact on a woman's health and quality of life, including an economic impact through loss of working hours. Treatment for chronic pelvic pain is often unsatisfactory. Present study compares Laparoscopic Uterosacral Nerve Ablation (LUNA) with laparoscopy without pelvic denervation in patients presenting with chronic pelvic pain to our outpatient clinic.Methods: It was a Randomised Controlled Trial Study. After considering inclusion and exclusion criteria, 120 patients were selected, out of which 60 (Group I) had undergone diagnostic laparoscopy and 60 (Group II) had undergone diagnostic laparoscopy with LUNA.Results: The overall success rate for group I and group II were 80%, 78.3% and 66.6% versus 85%, 81.6%, and 83.3% at 3, 6, and 12 months, respectively. However, on subgroup analysis it was found that in patients suffering from Congestive Dysmenorrhoea, there was a significant difference in success rate of both the groups.Conclusions: It was found in present study that there was a benefit for patients with dysmenorrheal, further research in this area is desirable to reach towards a discrete conclusion regarding the benefits of LUNA in patients of CPP.

Effect of casein phosphopeptide-amorphous calcium phosphate on fluoride release and micro-shear bond strength of resin-modified glass ionomer cement in caries-affected dentin

OBJECTIVES: This study was conducted to evaluate fluoride release and the micro-shear bond strength of resin-modified glass ionomer cement (RMGIC) in casein phosphopeptide-amorphous calcium phosphate (CPP-ACP)-remineralized caries-affected dentin (CAD). MATERIALS AND METHODS: Exposed dentin surfaces of 30 human third molar teeth were divided into 2 equal groups for evaluating fluoride release and the micro-shear bond strength of RMGIC to CAD. Each group was subdivided into 3 equal subgroups: 1) control (sound dentin); 2) artificially demineralized dentin (CAD); 3) CPP-ACP remineralized dentin (remineralized CAD). To measure fluoride release, 15 disc-shaped specimens of RMGIC (4 mm in diameter and 2 mm in thickness) were bonded on one flat surface of the dentin discs of each group. Fluoride release was tested using ion chromatography at different intervals; 24 hours, 3, 5, 7 days. RMGIC micro-cylinders were built on the flat dentin surface of the 15 discs, which were prepared according to the assigned group. Micro-shear bond strength was measured after 24 hours water storage. Data were analyzed using 1- and 2-way analysis of variance and the post hoc least significant difference test (α = 0.05). RESULTS: Fluoride detected in solutions (at all intervals) and the micro-shear bond strength of RMGIC bonded to CPP-ACP-remineralized dentin were significantly higher than those bonded to artificial CAD (p < 0.05). CONCLUSIONS: Demineralized CAD consumes more fluoride released from RMGIC into the solution for remineralization than CPP-ACP mineralized dentin does. CPP-ACP increases the micro-shear bond strength of RMGIC to CAD.

Expression of RGS4 and D2 receptor signaling pathway in striatum of methamphetamine-dependent CPP rats / 中国临床药理学与治疗学

AIM:To study the expression of signal transduction molecules in the striatum G protein protein 4 (RGS4) and dopamine D2 receptor (D2) in conditioned place preference (CPP) rats treated with methamphetamine (meth).METHODS:METH dependence CPP model was established (1 week and 2 weeks of METH dependence groups),The protein expression of RGS4 and D2,inhibitory G protein alpha-subunit (Gαi),mitogenactivated protein kinase (MAPK) in striatum were determined by Western blotting (WB).The changes of cyclic adenosine monophosphate (cAMP) content in striatum of rats were determined by enzyme linked immunosorbent assay (ELISA).RESULTS:Compared with saline control group,the average time of rats in the methamphetamine-paired chamber for two groups was increased (P ＜ 0.05).Compared with saline control group,RGS4 protein expressions in the two METH dependent groups were reduced (P ＜0.01);compared with 1 week of METH dependence group,that of 2 weeks group was reduced significantly(P ＜ 0.05).D2,Gαi,MAPK protein and cAMP expressions in the two METH dependent groups were increased (P ＜ 0.01);compared with 1 week of METH dependence group,those of 2 weeks were increased significantly (P ＜ 0.05).CONCLUSION:RGS4 and D2 receptor signaling pathways in striatum have changed in METH dependent rats,RGS4 may be involved in the regulation of METH-dependent D2 receptor signaling pathway in METH dependent rats.

Content Determination of Saccharide in Polysaccharides Containing Galacturonic Acid by Phenol-sulfuric Acid Method Combined with Calibration Factor Method / 中国药房

OBJECTIVE:To establish a method for the content determination of saccharide in polysaccharides containing galact-uronic acid based on phenol-sulfuric acid method combined with correction factor method. METHODS:The determination condi-tion of phenol-sulfuric acid was optimized. A series of standard curves were drawn with glacturonic aid-glucose mixed control with different mass ratio. The content of saccharide in Codonopsis pilosula polysaccharides CPP1b samples containing galacturonic acid was determined. According to regression equation of galacturonic acid-glucose ratio of 0-100%,the correction factor was calculated by using C. pilosula polysaccharides CPP1b as the reference polysaccharide,and the results of content determination of saccharide were corrected. The rationality of this method was verified by using mixed monosaccharide control with same composition as C. pi-losula polysaccharides CPP1b. RESULTS:The correction factor of C. pilosula polysaccharide CPP1b to glucose was 3.33;in vali-dation test,the content of saccharide in mixed monosaccharide control with same composition as C. pilosula polysaccharides CPP1b was 103.47%. RSDs of precision and stability tests was <1%. The recoveries ranged 93.52%-107.35%(RSD=5.09%,n=6). CONCLUSIONS:The established method can accurately determine the content of saccharide in C. pilosula polysaccharides CPP1b containing galacturonic acid.

Clinical evaluation of the preventing effect of 4 anti-caries preparations on enamel decalcification in orthodontic treatment with fixed appliance / 实用口腔医学杂志

Objective:To evaluate the preventing effects of 4 anti-caries preparations on enamel demineralization in fixed appliance orthodontic treatment.Methods:200 patients treated with fixed appliance were included and randomly divided into 4 groups (n =50),the teeth were treated with fluoride-containing toothpaste,fluoride varnish,fluoride free toothpaste and casein phosphopeptide-amorphous calcium phoaphate(CPP-ACP) respectively.The enamel decalcification incidence was calculated after orthodontic treatment.Results:The incidence of enamel decalcification calculated by tooth number was 16.3％ in fluoride-containing toothpaste group,9.3％ in fluoride varnish group,21.9％ in fluoride free toothpaste and 8.5％ in CPP-ACP group(among groups,P ＜0.05;between CPP-ACP and fluoride varnish,P ＞ 0.05).Conclusion:Fluoride containing preparation can prevent the enamel demineralization during the fixed appliance orthodontic treatment,CPP-ACP and fluoride varnish are more effective.

Effect of mPFC neuron synaptic plasticity changes in the formation of morphine related reward memory / 中华行为医学与脑科学杂志

Objective To observe the effect of mPFC neuron synaptic plasticity changes in the formation of morphine related reward memory.Methods 40 SD rats were administered morphine (10 mg/kg,ip) or saline (2 ml/kg,ip)and sacrificed 0,2,4 and 8 h after the treatment.The temporal profile of activity-regulated cytoskeleton-associated protein (Arc/Arg 3.1) expression in medial prefrontal cortex (mPFC) was analyzed.Another 40 rats receiving a single injection of morphine at different doses (0,5,10 or 20 mg/kg),and rats were sacrificed by decapitation 2 h later.In mPFC,changes of Arc/Arg 3.1 protein was analyzed by Western Blot,Arc/Arg 3.1 positive cells was detected by immunohistochemistry (IHC),and number of spines were analyzed by Golgi-cox method.In the second experiment,CPP model was established by 5 mg/kg morphine for 8 days.Arc/Arg 3.1 antisense oligodeoxynucleotide (AS) or the control (CS) was microinjected into mPFC 15 minutes before each morphine injection,then CPP score was evaluated.Results Compared with saline groups,Arc/Arg 3.1 protein,Arc/Arg 3.1 positive cells,number of spines ((1.01±0.04) vs (1.58±0.18),P＜0.01 ; (42.80±7.63) vs (74.47±8.02),P＜0.01 ;(17.27±5.64) vs (39.47±7.56),P＜0.01) were significantly increased 2 hours after morphine administration.All three doses of morphine (5,10 and 20 mg/kg) increased Arc/Arg 3.1 protein expression in the mPFC,and there were no dose-dependent effects.In CPP experiments,compared with microinjection of Arc/Arg 3.1 CS (0.74±0.02),Arc/Arg 3.1 AS microinjection significantly decreased the CPP score (0.51±0.01) in morphine group (P＜0.01).Conclusion It is enough to increase Arc/Arg 3.1 protein content and synaptic plasticity in mPFC by 10 mg/kg,and the changes implied in formation of morphine relative reward memory.

Casein phosphopeptide-amorphous calcium phosphate nanocom-plexes as a preventive agent for radiation caries and dental sensi-tivity in irradiated head and neck cancer patients / 中国肿瘤临床

Objective: To compare the efficacy of casein phosphopeptide-amorphous calcium phosphate nanocomplexes (CPP-ACP) and Duraphat varnish on preventing radiation caries and dentin hypersensitivity (DH) in irradiated head and neck cancer pa-tients. Methods:A total of 112 patients with head and neck cancer who will undergo radiotherapy were selected and randomly divided into the CPP-ACP group (experimental group) and the Duraphat varnish group (control group). A day prior to the scheduled radiothera-py, CPP-ACP containing paste had been applied once a day for 5 weeks to the teeth surface of the experimental group. Duraphat varnish was applied with the same frequency in the control group. The decay missing filling tooth (DMFT), decay missing filling surface (DMFS), and DH of two groups before radiotherapy and 12 months after radiotherapy were observed and statistically analyzed. Re-sults:Before radiotherapy, DMFT, DMFS, and DH showed no significant differences between the groups (P>0.05). However, signifi-cant increase in DMFT, DMFS, and DH was noted in the two groups 12 months after the radiotherapy (P<0.05). A higher increase was observed in the DMFT, DMFS, and DH of the control group compared with those in the experimental group (P<0.05). Conclusion:CPP-ACP effectively reduced radiation caries and sensitivity in irradiated head and neck cancer patients and deservesclinical applica-tion.

The effect of casein phosphopeptide amorphous calcium phosphate on the in vitro shear bond strength of orthodontic brackets / 대한치과교정학회지

OBJECTIVE: The purpose of this study was to evaluate the effect of casein phosphopeptide amorphous calcium phosphate (CPP-ACP) on the shear bond strength (SBS) of brackets bonded to non-demineralized teeth with either phosphoric acid etching or self-etching primer. METHODS: Sixty human premolars were randomly assigned to 1 of 4 treatment groups (n = 15 each): phosphoric acid etching (group 1); self-etching primer (group 2); CPP-ACP for 2 weeks + phosphoric acid etching (group 3), and CPP-ACP for 2 weeks + self-etching primer (group 4). After bonding of the maxillary premolar metal brackets, specimens were subjected to shear forces in a testing machine. Scanning electron microscopy was used to observe etching patterns on the enamel surfaces of all teeth. A 2-way analysis of variance was used to test for effects of CPP-ACP and etching system on SBS. RESULTS: Significantly higher mean SBSs were observed in groups subjected to phosphoric acid etching (i.e., groups 1 and 3; p 0.05). We observed a uniform and clear etched pattern on the enamel surface of the phosphoric acid etching groups. CONCLUSIONS: CPP-ACP does not significantly affect the SBS of orthodontic brackets bonded to non-demineralized teeth, regardless of which adhesive method is used to bond the brackets.

Functionality advancement of poorly soluble biovariable anti hypertensive Drug by sophisticated SD-FBP technology as per enhanced QbD.

Lacidipine (LCDP) is a dihydropyridine derivative categorized as an Anti-hypertensive Ca+2 channel blocker belonging to BCS class IV drug with low solubility and low permeability which presents a challenge to the formulation scientists. The development of a solid dispersion by solvent evaporation is a practically viable method to enhance dissolution of LCDP from oral dosage form. Solvent evaporation by Fluidized Bed Process (FBP) was the method of choice for SD as it improves wettability with simultaneous increase in porosity of granules resulting enhanced surface area producing higher dissolution rate and bioavailability of poorly water-soluble drug. Thus, the main object of the present invention is to provide stable pharmaceutical dosage form of LCDP with desired dissolution rate i.e. at least 80% drug release within 45 minutes, without use of disintegrant(s) and/or surfactant(s) or without micronization of the active ingredient per se. One more object of this invention is to provide a sophisticated robust process for the preparation of said pharmaceutical dosage form by Quality by Design (QbD) concept focusing on thorough understanding of the product and process by which it is developed and manufactured along with a knowledge of the risks involved in manufacturing by IRMA & FMEA study of the product with process and how best to mitigate those risks by developing design space with DoE & MVDA with outlined control strategy.

The Role of PKMzeta in Drug Reward Memory / 대한정신약물학회지

Drug addiction is a chronic brain disease with a high incidence of relapse. Environmental cues that previously and repeatedly associated with drugs of abuse easily evoke relapse to addicts even after long period of drug-free state. Such a long lasting property of conditioning is considered a form of long-term memory and has a strong correlation with synaptic plasticity like long-term potentiation (LTP). Protein kinase M zeta (PKMzeta) has been known to play an important role in the maintenance of long-term memory as well as LTP in various brain areas. Likewise, in a few brain areas examined out of the rewarding circuit, PKMzeta seems to play a similarly important role in the maintenance of conditioned memory. These results suggest that PKMzeta may become a new target to manipulate to reverse pre-formed drug-related memory and accompanied behaviors.

Efficacy of CPP-ACP and CPP-ACPF on enamel remineralization - An in vitro study using scanning electron microscope and DIAGNOdent®.

Introduction: Remineralization as a treatment procedure has received a lot of attention both from clinicians as well researchers. The objective of this in vitro study was to find out the efficacy of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) and casein phosphopeptide-amorphous calcium phosphate fluoride (CPP-ACPF) in remineralizing enamel surface on which artificial caries lesion had been created. The changes were analyzed using DIAGNOdent® (KaVo) and scanning electron microscope (SEM). Materials and Methods: Ninety maxillary premolars were selected and divided into three groups of 30 teeth each: A (artificial saliva), B (CPP-ACP), and C (CPP-ACPF). All the samples were assessed using DIAGNOdent® at the baseline and after demineralization and remineralization. Three samples were randomly selected from each group after remineralization for surface evaluation using SEM. Results: Statistical analysis showed that group B {CPP-ACP (4.1±1.8)} and group C {CPP-ACPF (4.8±1.2)} had a significantly higher amount of remineralization than group A (1.7±0.7). Conclusion: All the three groups showed a statistically significant amount of remineralization. However, because of the added benefit of fluoride (NaF 0.2%), CPP-ACPF (Tooth Mousse-Plus®) showed marginally more amount of remineralization than CPP-ACP (Tooth Mousse®).

Immunoregulatory effects of Periplocin from Cortex Periplocae in tumor-bearing mice / 细胞与分子免疫学杂志

AIM: To investigate the immune mechanisms for Periplocin from Cortex Periplocae (CPP) in tumor-bearing mice. METHODS: H_(22) tumor-bearing model BALB/c mice were applied to evaluated in vivo immunoregulatory effect of CPP. The influence of different dose CPP (0.25, 0.50 and 1.00 mg/kg) on immune organs in tumorbearing mice were observed. T cell subsets of mice spleen were detected by flow cytometry. MTT assay was used to determine the influence of CPP on lymphocyte proliferation of mice spleen stimulated by ConA. The levels of TNF-α, IL-2 and IL-12 in serum from mice were detected by means of ELISA. RESULTS: Thymus index and spleen index of H_(22) tumor-bearing model control mice became less than that of normal mice (P < 0.05). Compared to both model and normal control groups, thymus index and spleen index of H_(22) tumor-bearing mice treated with CPP increased obviously (P < 0.05). CPP had no influence on the number of CD8~+ T cells, but up-regulated markedly the number of CD3~+, CD4~+ T cells and the ratio of CD4~+/CD8~+ in tumorbearing mice. In CPP-treated mice, the percentage of CD3~+, CD4~+ T cells were not different from normal mice (P<0.05), the ratio of CD4~+/CD8~+ was higher than that of normal mice (P < 0.05). CPP enhanced obviously lymphocyte proliferation of mice spleen induced by ConA, the SI scores were even higher than that of nornal mice. The levels of TNF-α, IL-2 and IL-12 in serum from CPP-treated mice, increased significantly compared to model control group (P<0.05) in a dose-dependent manner, were similar to or higher than that of normal mice. CONCLUSION: CPP protected immune organs of tumor-bearing mice, increased obviously the percentage of CD4~+ and CD4~+/ CD8~+ among the T cell line, and enhanced lymphocyte proliferation of mice spleen significantly, stimulated the production of TNF-α, IL-2 and IL-12. The results suggested that CPP possessed potent immunoregulatory effect.

The Electron Microscopic Study on the Development of Knee Joint in Rat / 대한해부학회지

These study was designed to observe the appearance and the characteristics of apoptotic cells during the development of knee joint in rat. The fetus were collected on the 16th, 17th, 18th, 19th, and 20th day of pregnancy. In this study, TUNEL staining, electron microscopic investigation and immunocytochemical gold labeling techniques were used. In the immuno-cytochemical gold labeling techniques, primary antibodies were used, which were to be polyclonal rabbit anti-mouse/ rat Bax, polyclonal rabbit anti-tissue transglutaminase C, and polyclonal goat anti-cpp32p20. The samples were observed under JEOL 1200 EX-II transmission electron microscope. The results were as follows. 1. In a 16-day-old fetus, between femur and tibia cartilages, mesenchymal cells were observed. Mesenchymal cells had marginated heterochromatin and dilated rough endoplasmic reticulum. 2. In a 17-day-old fetus, the knee joint clefts were first formed. In the primordial cruciate ligaments between the cartilages, capillaries were scattered. The apoptotic cells, which had fragmented and condensed nucleus, showed in the synovium. And necrotic cells, which had nuclear chromatin margination, perinuclear cisternae, and dilated rough endoplasmic reticulum, also were observed in the joint cleft surface. 3. From the 18-day-old fetus, phagocytic synovial cells and secretory synovial cells could be confirmed. The apoptotic cells were not seen. 4. In a 17-day-old fetus, a few cells were positive for TUNEL reaction in the joint cleft region. 5. In a 17-day-old fetus, Bax were marked on the mitochondria, endoplasmic reticulum of apoptotic cells. Also, it was marked at the phagocytosed apoptotic bodies in the neighboring cells. 6. In a 17-day-old fetus, the tissue Transglutaminase C were marked in the perinuclear region, vacuoles, cell membrane and extracellular matrix of the apoptotic cells. Also, it was marked at the phagocytosed apoptotic bodies in the neighboring cells. 7. In a 17-days-old fetus, CPP32 labeling were marked in the cytoplasm of the apoptotic cells. Practically, it was distributed between the phagocytosed apoptotic bodies and the neighboring cells. On the basis of above findings, it is obvious that the joint cleft are first formed in a 17-day-old fetus, a few cells are to be TUNEL positive signals, and the apoptotic cells contain Bax, tissue Transglutaminase C, and CPP32. Therefore the apoptotic cells and the necrotic cells are appeared in the 17-day-old fetus, and these cells are concerned with joint cleft formation.

CLONING AND EXPRESSION OF A GENE MEDIATINGγ-RADIATION-INDUCED APOPTOSIS IN HL-60 CELLS / 药物分析学报

Objective To identify the member of the caspase family proteases involved in γ-radiation-induced apoptosis in HL-60 cells and to study the expression of the caspase gene in normal, apoptotic cells and in immortal tu mor cells. Methods By using degenerate oligonucleotide primers encoding the highly conserved peptides that were pre sent in all known caspases, we performed RT-PCR on poly(A)RNA from γ-radiation-induced apoptotic HL-60 cells. Caspase-3 mRNA in apoptotic HL-60 cells and in human tumor cell lines was analyzed by Northern blot. Results The amplified DNA fragment was identified with caspase-3 cDNA by cloning and sequencing. The Northern blot analysis of caspase-3 mRNA of different human tumor cell lines showed that the caspase-3 gene transcript was more highly ex pressed in leukemia cell lines and the SH-SY5Y cell line than in HeLa and MCF-7 cells. It was more highly expressed in the radiation-induced apoptotic HL-60 cells than in control HL-60 cells. Conclusion These results indicated that caspase-3 was involved in γ-radiation-induced apoptosis in HL-60 cells. The high level of expression of caspase-3 may aid efforts to understand the insensitivity of some tumor cells to radiation, their inherent ability to survive, and apop tosis.

Cell Beath Induced by Ethanol : Prevention of Cell Death by the bcl-2 Proto-Oncogene

The Bcl-2 protein has been shown to block apoptosis induced by a variety of stimuli. We have performed the experiments which cell death can be blocked by the bcl-2 proto-oncogene under moderate(50-100mM) or high ethanol treatment(400-600mM). As a result of morphological changes, and MTT assay, cell death was blocked by Bcl-2 under 100mM ethanol. However, the results of DNA fragmentation and RT-PCR(ICE, and CPP32), immunoblotting(CPP32, and PARP) for SK-pcDNA3 cells(vector only) and SK-Bcl-2 cells(stably expressed bcl-2 gene) were showen to be no significant differences between two cell lines. These result suggested that cell death induced by ethanol was not followed by apoptosis mechanism, and was blocked by the bcl-2 proto-oncogene with moderate ethanol.

Antinociceptive Effects of Intrathecal Morphine and NMDA Receptor Antagonists, MK 801 and CPP, on the Formalin Test in Rats / 대한마취과학회지

BACKGROUND: It is emphasized that repetitive stimulation of small diameter afferent fibers produces a progressive increase in the action potential discharge and a prolonged increase in the excitability of neurons in the spinal cord following the stimulus and that this facilitatory component has a unique pharmacology. To investigate the behavioral parallels of this spinal facilitation, we evalusted the antinociceptive effects of intrathecal morphine, N-methyl-D-aspartate(NMDA), (+)-5-methyl-10,11- dihydro-5H-dibizo(a,d) cycloheptene-5, 10-imine hydrogen maleate(MK801) and (+/-)-3-(2-carboxy- piperazine-4-yl)-propyl-I-phosphonic acid(CPP), on the formalin test in rats. METHODS: Four to six days after chronic lumbar intrathecal catheterization, normal saline, morphine(0.1 to 30 ug), MK801(0.1 to 10 ug), CPP(0.1 to 5 ug) or NMDA(10 or 100 ng) were administered intrathecally before formalin injection. Spontanesous flinches were observed at 1-2 and 5-6 min(phase 1) and at 10 min intervals thereafter for 50 min(phase 2) after subcutaneous formalin injection into the dorsum of the right hind paw for each drug treated rats. RESULTS: Intrathecal morphine produced dose dependent inhibition of the phase 1 and phase 2 response(ED50=0.63 ug and 0.37 ug, respectively). Intrathecal MK801(0.1 to 10 ug) and CPP(0.1 to 5 ug) inhibited the phase 2 response more strongly than phase 1 response and inhibition of the phase 2 response(P0.7). Relative potencies of MK801 and CPP when compared with morphine were 1.34 and 0.41, respectively. CONCLUSIONS: This study suggests that intrathecal morphine and NMDA receptor antagonist(MK801 and CPP) have an antinociceptive effect on pathological pain mediated by central sensitization and that NMDA receptor antagonists can be utilized selectively in the treatment of components of central sensitization.

Effect of radix of Salvia miltiorrhiza fraction on conditioned place preference in mice induced by morphine / 中草药

Objective To investigate the effect of lipid soluble fraction in the radix of Salvia miltiorrhiza (FSM) on conditioned place preference (CPP) in mice induced by morphine and preliminarily identify the fraction in the radix of S. miltiorrhiza. MethodsMorphine or NS was sc injected every other day to induce the obvious CPP in mice for 6 d. Before 30 min of sc injecting morphine, mice were ip administered different doses of lipid soluble fraction in the radix of S. miltiorrhiza. RP-HPLC method was used to identify the major component in the lipid soluble fraction in the radix of S. miltiorrhiza. ResultsThe staying time in morphine-paired white compartment was significantly prolonged. After treatment with lipid soluble fraction in the radix of S. miltiorrhiza (40 mg/kg, ip), the staying time in morphine-paired white compartment was significantly shortened (P

Clinical analysis of 34 diffuse axonal injured (DAI) patients below GCS 8

A consecutive series of 34 severe head-injured patients (DAI) were studied prospectively. Patients were categorized according to a new, simple classification system comprised of four lesion types according to the compression or obliteration of the ventricles or cisterns. Five patients belonged to type II and 19 patients to type IV. Each type was further subdivided into two GCS score ranges (5 to 8 and below 5). The distribution of the posttraumatic infarction was mainly in the frontal and temporal lobes (60% of all cases). Our data demonstrated that the ICP was significantly lower at a 30 degrees head elevation than at 0 degree (18.6 +/- 7.21 mmHg vs 23.0 +/- 10.60 mmHg. t = 4.22 p< 0.001), but head position did not statistically affect CPP (69.4 +/- 19.86 mmHg vs 68.2 +/- 19.87 mmHg. t = -0.54, p< 0.59). The effect of intensive therapy on ICP, CPP and AVDO2 was studied in all cases, employing steroids and diuretics in a modified intensive care scale. In cases where barbiturates were employed, there were statistically significant changes in ICP and AVDO2 (p< 0.001), but CPP was not affected (p< 0.59). Surviving patients were analyzed by using the GOS and the neurological grading score (NGS, Nihon University) of the persistent vegetative state. Our data suggests that head elevation of 30 degrees and barbiturate therapy are more effective on ICP and AVDO2, and NGS more exact than GOS in vegetative patients.