Objective To study the different expressions of receptor activator of nuclear factor-kappa B ligang(RANKL) mRNA in spleens of rats fed with diet of low calcium and high fluoride. Methods A 2× 2×2 factorial design was used and the factors were calcium, fluoride and action time. In the design, 40 Wistar rats [average body mass(118.9±13.5)g] were divided into four groups randomly by weight: control with normal diet (0.790%, calcium), low calcium group with low calcium intake(0.063%, calcium), high fluoride group with normal diet and high fluoride intake(100 mg/L, fluoride) and low calcium and high fluoride group with low calcium and high fluoride intake. After 4 and 8 months, 5 rats of each group were sacrificed and total RNA was extracted from spleen. And the expression levels of RANKL mRNA were determined by reverse transcription polymerase chain reaction (RT-PCR). Results At time of 4 months, the expression level of RANKL mRNA was 0.13± 0.05,0.13± 0.03,0.17±0.02,0.27± 0.05 and at time of 8 months, it was 0.11 ± 0.01,0.16 ± 0.02,0.16± 0.03,0.36 ± 0.07 in control group, low calcium group, high fluoride group, low calcium with high fluoride group, repectively. The factorial design AVONA showed that low calcium and high fluoride had significant effects on RANKL mRNA expression(F = 40.224,56.679, all P < 0.05) while action time had not(F = 2.850, P > 0.05 ). The interactions of low calcium with high fluoride or high fluoride with action time were signifieant(F = 7.247, 18.789, all P < 0.05) while the interaction of high fluoride with action time was not(F = 1.751, P > 0.05). Conclusions Low calcium alone or high fluoride alone or low calcium with high fluoride or low calcium with action time can increase the the RANKL mRNA expression level. High fluoride does not affect the RANKL mRNA level as the action time is prolonged.

To systematically review the incidence rate of adverse drug reaction/event by Qingkailing injection. Such databases as the PubMed, EMbase, the Cochrane library, CNKI, VIP WanFang data and CBM were searched by computer from foundation to July 30, 2015. Two reviewers independently screened literature according to the inclusion and exclusion criteria, extracted data and cross check data. Then, Meta-analysis was performed by using the R 3.2.0 software, subgroup sensitivity analysis was performed based on age, mode of medicine, observation time and research quality. Sixty-three studies involving 9,793 patients with Qingkailing injection were included, 367 cases of adverse reactions/events were reported in total. The incidence rate of adverse reaction in skin and mucosa group was 2% [95% CI (0.02; 0.03)]; the digestive system adverse reaction was 6% [95% CI(0.05; 0.07); the injection site adverse reaction was 4% [95% CI (0.02; 0.07)]. In the digestive system as the main types of adverse reactions/events, incidence of children and adults were 4.6% [0.021 1; 0.097 7] and 6.9% [0.053 5; 0.089 8], respectively. Adverse reactions to skin and mucous membrane damage as the main performance/event type, the observation time > 7 days and ≤ 7 days incidence of 3% [0.012 9; 0.068 3] and 1.9% [0.007 8; 0.046 1], respectively. Subgroup analysis showed that different types of adverse reactions, combination in the incidence of adverse reactions/events were higher than that of single drug, the difference was statistically significant (P < 0.05). This study suggested the influence factors of adverse reactions occur, and clinical rational drug use, such as combination, age and other fators, and the influence factors vary in different populations. Therefore, clinical doctors for children and the elderly use special care was required for a clear and open spirit injection, the implementation of individualized medication.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Drugs, Chinese Herbal ; adverse effects ; Female ; Humans ; Incidence ; Infant ; Infant, Newborn ; Injections ; Male ; Middle Aged

Recombinant human BMP-2 was compounded with chitosan/gelatin/hydroxyapatite(HCG) scaffold and the complex was sterilized by 60Co radiating. Osteoblast isolated from cranial bones of newborn rat was primary cultured and seeded onto the complexes. 3 days after culturing, scanning electron microscope(SEM) was applied to detect the compatibility of the cell with the complex. SEM showed osteoblast attached closely with the complex and grew well in its pores. Then the complexes with osteoblast modification were implanted into athymic nude mice subcutaneously. 8 weeks after implantation, X-ray photograph and histological observation were applied to detect the bone formation of the complexes. Under X-ray a high-density areas consistent with the shape of the implanted complex could be seen. Histological observation also proved there was bone formation in the interspace of the complex. A conclusion was drawn that rhBMP-2 compounded HCG scaffold had good osteogenesis ability in vivo.

Objective:To analyze the relationship between hepatitis B virus(HBV)gene mutation at 1896 in precore region with genotype and replication of HBV and the liver function of patients.Methods:HBV precore 1896 site mutation,the genotype of HBV and serum content of HBV DNA were determined by PCR in 60 patients positive of HBV DNA.Chemiluminescence miacropaticle immunoassay(CMIA)was used for detection of serum HBeAg and HBeAb.Liver function parameters were ob- tained by routine biochemistry method.Results:The alanine aminotransferase(ALT)level in HBV with 1896 site mutation was significantly higher than that in the wildtype virus.Site mutation at 1896 had no correlation with HBeAg,HBV genotype and HBV DNA content.HBV DNA content in patient with genotype C was significantly higher than that with genotype B(P

Adult ; Aged ; Carcinoma, Squamous Cell ; pathology ; virology ; Cervical Intraepithelial Neoplasia ; pathology ; virology ; Colposcopy ; Cytodiagnosis ; DNA Probes, HPV ; DNA, Viral ; isolation & purification ; Female ; Humans ; Middle Aged ; Nucleic Acid Hybridization ; Papillomaviridae ; genetics ; isolation & purification ; Papillomavirus Infections ; Uterine Cervical Neoplasms ; pathology ; virology ; Vaginal Smears ; Young Adult

Objective:To observe the clinical efficacy and safety of cranial suture acupuncture plus paroxetine in treating depression,and to discuss the action mechanism of this acupuncture method.Methods:One hundred depression patients were allocated to an observation group and a control group according to the random number table,with 50 cases in each group.The control group was intervened by oral administration of paroxetine tablets,20 mg each time,once a day for successive 6 weeks;the observation group was additionally given cranial suture acupuncture,once a day for 6 weeks.They were scored by Hamilton depression scale-17 (HAMD-17) before the treatment and respectively after 1-week,2-week,4-week and 6-week treatment.The clinical efficacy and safety were also observed.Results:After 6-week treatment,the total effective rate was 94.0％ in the observation group versus 78.0％ in the control group,and the between-group difference was statistically significant (P＜0.01).The HAMD-17 scores respectively after 1-week,2-week,4-week and 6-week treatment were significantly lower than the score before the treatment in the observation group (all P＜0.05);the HAMD-17 scores respectively after 2-week,4-week and 6-week treatment were significantly different from the score before the treatment in the control group (all P＜0.05).There were significant differences in the HAMD-17 score between the two groups respectively after 4-week and 6-week treatment (both P＜0.05).Conclusion:Cranial suture acupuncture plus paroxetine can ease the symptoms of depression,with faster onset and more significant therapeutic efficacy compared with paroxetine alone.

AIM AND METHODSThe properties and sensitivity to acetylcholine of PC12 cells differentiated with nerve growth factor (NGF) have been investigated by using whole-cell clamp technique.

RESULTSWhen cultured in the presence of NGF, PC12 cells not only differentiated to resemble sympathetic neurons morphologically, but also developed electrical excitability. NGF-treated PC12 cells were highly sensitive to ACh than untreated cells. The I(Ach) proved to be generated by nAChR by pharmacological identification. Nicotinic receptor was characterized by desensitization. The macroscopic I(ACh) was inward rectified and concentration dependent.

CONCLUSIONPC12 cells are easily cultured and provides a homogenous population of cells. When culture in NGF, they differentiate to sympathetic-like neurons that contain on their surface neuronal nAChR, it can be used as good model system for studying regulation of a sympathetic neuronal nAChR.

Acetylcholine ; pharmacology ; Animals ; Cell Differentiation ; Nerve Growth Factor ; metabolism ; Neurons ; metabolism ; PC12 Cells ; Patch-Clamp Techniques ; Rats ; Receptors, Nicotinic ; metabolism

The mechanism of long-term potentiation (LTP) in basolateral amygdala (BLA) was explored using field potential recording in rat brain slice preparation. Field potentials (field excitatory post-synaptic potentials, fEPSPs) in BLA were evoked with sharpened steel bipolar stimulating electrodes placed in the external capsule (EC). Two theta burst stimulations (TBS, interval=10 min) induced LTP in BLA. TBS-induced synaptic potentiation lasted for more than 30 min after the second TBS. LTP in BLA was input-specific and was blocked by N-methyl-D-aspartate receptor (NMDAR) antagonist 2-amino-5-phosphonovaleric acid (APV). The effect of protein kinase C (PKC) on LTP was then determined using PKC inhibitor chelerythrine chloride. Bath application of chelerythringe chloride had no effect on basic field potentials and paired-pulse ratio (PPR). However, in the presence of chelerythrine chloride, two TBS failed to induce LTP. In contrast, bath application of chelerythrine chloride 10 min after the second TBS did not affect the maintenance of LTP in BLA. These results indicate that LTP is NMDAR-dependent and PKC is involved in the induction and early maintenance of LTP in BLA.
2-Amino-5-phosphonovalerate ; pharmacology ; Amygdala ; enzymology ; Animals ; Electric Stimulation ; In Vitro Techniques ; Long-Term Potentiation ; Protein Kinase C ; metabolism ; Rats ; Receptors, N-Methyl-D-Aspartate ; metabolism ; Synaptic Potentials

OBJECTIVETo investigate the effects of growth factors on c-fos and c-jun gene expressions in hepatic stellate cells.

METHODSHepatic stellate cell (HSC) T6 was cultured in media containing different concentrations of platelet derived growth factor (PDGF) (8 ng/ml, 40 ng/ml or 200 ng/ml) and transforming growth factor (TGF) beta (0.2 ng/ml, 1 ng/ml or 5 ng/ml) and the cells were collected at different incubation periods (8, 24, 48 or 72 h). Total RNA of the HSC was isolated and c-fos and c-jun gene expression levels were measured by reverse transcription polymerase chain reaction.

RESULTSC-fos gene expression levels of the HSC cultured in low (8 ng/ml), medium (40 ng/ml) and high (200 ng/ml) concentrations of PDGF were all much higher than those of the control group after exposure to PDGF at 8, 24, 48 or 72 h. The c-fos gene expression levels of the HSC increased as the dosage of PDGF increased and there were significant differences of c-fos gene expression among the three PDGF groups. C-jun gene expression levels of the HSC in low (0.2 ng/ml), medium (1.0 ng/ml) and high (5.0 ng/ml) concentrations of TGF beta groups were much higher than those of the control group after exposure to TGF beta at 8, 24, 48 or 72 h. The c-jun gene expression levels of the HSC increased as the dosage of TGF beta increased and there were significant differences of c-jun gene expression among the three TGF beta groups.

CONCLUSIONPDGF and TGF beta can strongly up-regulate c-fos and c-jun gene expressions in hepatic stellate T6 cells.

Cells, Cultured ; Gene Expression ; Hepatic Stellate Cells ; drug effects ; metabolism ; Humans ; Platelet-Derived Growth Factor ; pharmacology ; Proto-Oncogene Proteins c-fos ; genetics ; Proto-Oncogene Proteins c-jun ; genetics ; Transforming Growth Factor beta ; pharmacology

To investigate the effects of gambognic acid (GA) on TRAIL-induced apoptosis of cancer cells, human colon HT-29 cancer cells were treated with GA to promote apoptosis. Inhibition of the cell proliferation was measured with MTT assay and cell apoptosis was detected with formation of DNA ladders in agarose gel electrophoresis, and activation of caspase activity. The content of cytosolic reactive oxygen species (ROS) was measured with flow cytometry. The activities of Caspase-3, -8, -9 were detected using spectrophotometric assay. The levels of c-FLIP, CHOP, DR4 and DR5 in cells were tested by Western blot. Combination of GA (1 µg · mL(-1)) and TRAIL (40 ng · mL(-1)) significantly reduced proliferation and increased apoptosis of HT-29 cells over those induced by each agent alone. Percentage of apoptotic cells was increased to 45.5%. GA markedly enhanced the intracellular ROS generation. Expression of CHOP, DR4 and DR5 was up-regulated to 7.38, 5.41, and 4.85 times of the control group, respectively. GA promoted activation of Caspase-3, -8, and -9 by TRAIL (P<0.05). Furthermore, the expression of anti-apoptotic protein c-FLIP was down-regulated to 0.22 ± 0.08 times of the control group. In conclusion, GA sensitizes HT-29 cells to TRAIL-induced apoptosis by promoting ROS-activated ERS pathways, up-regulating of DR4 and DR5, and inhibiting c-FLIP expression.
Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Caspases ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; metabolism ; Down-Regulation ; HT29 Cells ; Humans ; Reactive Oxygen Species ; metabolism ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; Up-Regulation ; Xanthones ; pharmacology