Objective To histomorphometricly assess changes occurred in the alveolar ridge following different methods of socket preservation and to compare them against natural healing without interventions. Methods The second、 third and fourth mandibular premolars were extracted from six beagles. Six extraction sites in each dog were randomly assigned to three treatments as follows:natural healing (T1), Bio-Oss Collagen (T2) and immediate implant with Bio-Oss (T3). Six month after surgery, the dogs were euthanized and tissue samples were sectioned, fixed and mounted, then were stained with toluidine blue. The histologic studies and morphological measurements were performed by using an optical microscope and a digital image software. Results Reabsorption in the buccal aspect of the alveolar crest of ridge was showed in all groups. With respect to the mean vertical bone loss of the buccal bone plate, T3 is lower than T1 and T2(P<0.001 ), while no significant differences were observed between T1 and T2. With regard to horizontal dimension of the alveolar process , a statistical significance could be found at 3mm and 4mm below the crest of ridge in group T1 and T3(P=0.017, P=0.042), while no statistical differences were found between other groups. Conclusions Both techniques of alveolar ridge preservation were not able to completely preserve the original bone volume after tooth extraction. Immediate implant placement in combination with Bio-Oss seems to have the potential to limit the reabsorption of the alveolar process efficiently , but the bone preserving effect of Bio-Oss Collagen is undesirable.

Constructing prokaryotic expression vector pKY-RMBAY by gene recombination and research its optimizing productive conditions.By PCR technology synthesizing the gene of the RMBAY with preference codon of E.coli and the RMBAY gene was inserted into high efficiency expression vector pKYB-MCS.Expressed fusion proteins in E.coli ER2566 were purified with Chitin-Beads column.Fusion proteins binding on Chitin-Beads was cut on N-terminus of intein due to the induction of ?-mercaptoethanol and the target peptide RMBAY was released.The RMBAY was identified by mass spectrum.Experiment results showed RMBAY can be high efficiently expressed in E.coli ER2566,with optimizing productive conditions the yield of the RMBAY may be 6.7mg/L fermentation product and its purity is greater than 98%.The molecular weight of RMBAY is 3.887 kDa by mass spectrum and that accords with its theory value.

In order to prepare the recombinant vasoactive intestinal peptide (VIP) using intein mediated rapid purification system,the cDNA encoding the recombinant VIP was designed and synthesized according to the preference of E.coli,and then was cloned into the expression vector PTWIN. The recombinant plasmid PTWIN-VIP was transformed into expression host E.coli strain ER2566.The fusion protein consisting of the recombinant VIP,intein and chitin binding domain was expressed and purified by chitin affinity chromatography. The target peptide was released from the fusion protein by changing the temperature and the pH of the cleavage buffer. The molecular weight of the recombinant VIP was determined by the mass spectrometry and the results was conformity with the theoretical value. The preliminary bioactivity assay indicated that the recombinant VIP decreased the serum resistin levels significantly in LPS-induced acute inflammation. The preparation and the characterization of anti-inflammatory effects of the recombinant VIP layed the foundation for its further application.

Objective To explore the effect of excessive fluoride on expression of mRNA and protein of Wnt3a and β-catenin in rats' osteoblasts and its correlation with pathogenic mechanism of fluorosis.Methods Thirty-six healthy SD rats,weighting 100-120 g and according to body mass,were randomly divided into three groups(twelve in each group).The rats of control were fed wich tap water(fluoride ＜ 1 mg/L) and the experimental rats were exposed to NaF(low-fluoride group:5 mg/L,high-fluoride group:50 mg/L) added to the drinking water to establish the chronic fluorosis model.After fed for eight morth,all rats were killed and metaphysic of femoral was collected.Rat dental fluorosis was observed and bone fluorine was detected by ashing-fluorin ion selective electrode method.The content of bone alkaline phosphatase (BALP) and tartrate-resistant acid phosphatase 5b(TRACP 5b) in rats' serum was detected by enzyme-linked immunosorbent assay (ELISA).The morphologic changes of the bone were observed by microscopy.The expression of mRNA and protein of Wnt3a and β-catenin in osteoblasts of rats was analyzed with gray scale by hybridization in situ and immunohistochemistry methods,respectively.Results Detection rate of dental fluorosis,fluoride contents of urine and bone were significantly increased [control group:0.0％,(1.26 + 0.17)mg/L,(305.58 ± 91.26)mg/kg; low-fluoride group:66.7％,(2.06 ± 0.64)mg/L,(632.33 ±123.21)mg/kg; high-fluoride group:91.7％,(7.69 ± 1.96)mg/L,(1088.75 ± 156.16) mg/kg] in the rats treated with fluoride,the difference between groups was statistically significant(χ2 =21.6; F =36.57,467.02; all P ＜0.05).The contents of BALP and TRACP-5b in rats' serum were significantly different between groups(F =89.57,7.68; all P ＜ 0.05).Compared with control group[(16.24 + 1.57)U/L],the contents of BALP in rats' serum of the low-fluoride and high-fluoride groups[(31.47 ± 5.30) and (54.61 ± 2.27)U/L] were increased gradually(all P ＜0.05).Compared with the low-fluoride group,the value in the high-fluoride group decreased significantly (P ＜ 0.05).The contents of TRACP-5b in rats' serum of low-fluoride group[(3.45 ± 1.85)U/L] were elevated significantly(all P ＜ 0.05) compared with the control group[(1.26 ± 0.23)U/L] and the high-fluoride group[(2.74 ± 1.85)U/L].The bone cortices were thickened and the bone trabecula was broadened,arranged closely together in chronic fluorosis rats with significant difference compared with the control group.In the low-fluoride and high-fluoride groups,the expression levels of Wnt3a and β-catenin mRNA (low-fluoride group:132.87 ± 5.72 and 132.57 ± 9.56; highfluoride group:135.60 ± 6.64 and 137.87 ± 9.16) were markedly elevated with significant difference,respectively (F =12.47,5.96; all P ＜ 0.05) compared with those in control groups(119.86 ± 5.04 and 120.58 ± 7.84) by hybridization in situ(P ＜ 0.05),but there was no statistical significance (P ＞ 0.05) of the level of Wnt3a and β-catenin mRNA between low-fluoride and high-fluoride groups.In the low-fluoride and high-fluoride groups,the protein expression of Wnt3a and β-catenin (low-fluoride group:137.50 ± 4.32 and 140.85 + 3.54; high-fluoride group:142.65 ± 11.84 and 152.52 ± 4.64) were markedly elevated with significant difference,respectively (F =10.07,53.82; all P ＜ 0.05) compared with those in control group (124.01 ± 2.63 and 126.75 ± 4.65) by immunohistochemistry(all P＜ 0.05),Wnt3a protein production in the low-fluoride group was increased without statistical significance compared with the high-fluoride group (P ＞ 0.05).But the protein production of β-catenin in the lowfluoride group was elevated with significant difference compared with the high-fluoride group(P ＜ 0.05).The mRNA and protein production of Wnt3a were positively correlated with the mRNA and protein production of β-catenin (r =0.731,0.658; all P ＜ 0.05).Conclusions Rat bone tissue lesions caused by excessive fluoride may be associated with an increased expression of Wnt3a and β-catenin mRNA and protein in osteoblasts.In chronic fluorosis,fluoride stimulates the overexpression of Wnt3a and β-catenin in the Wnt signal transduction pathway,enhances bone osteogenesis and causes skeletal fluorosis.

Objective To investigate the value of the hippocampal subfield's MRI T2 signal intensity in evaluating the effect of the hydrochloric donepezil for mild cognitive impairment(MCI).Methods 20 MCI patients with hydrochloric donepezil (treatment group) and 20 patients with placebo (control group) were scanned by MRI using FSE-T2 sequence.The margin of hippocampal subfields was outlined manually for each side to measure the MRI T2 signal intensity.The relationship between hippocampal MRI T2 signal intensity and MMSE scores was analyzed in the treatment group.Results Before the treatment, there was no significant difference of the MRI T2 signal intensity between groups.After the treatment, the MRI T2 signal intensity in the bilateral head of the treatment group was significantly decreased compared with the control group(P<0.05).There was an inverse relationship between the MRI T2 signal intensity of the bilateral hippocampal head and MMSE scores in the treatment group(P<0.05).Conclusion The MRI T2 signal intensity in the bilateral hippocampal head could be regarded as a valuable marker in making clinical diagnosis and evaluating the effect of the treatment for MCI in its early stage.

Objective To analyze the effect of N-acetyl-L-cysteine(NAC)on endoplasmic reticulum stress mediated HepG2 cells apoptosis and evaluate the role of NAC in the treatment of liver injury.Methods HepG2 cells were treated with thapsigargin(TG)to establish the model of oxidative endoplasmic reticulum stress mediated apoptosis,and NAC was used to intervene in apoptosis.To evaluate the apoptosis,various methods such as MTT assay,flow cytometry,DNA ladder and Western blot were performed.Results After treated with 2 μmol/L TG for 0,24,36 and 48 hours,the vitality of HepG2 cells decreased.The ratio of apoptotic cells increased along with the prolonged treatment duration of TG(0.7%±0.5%,27.6%±6.3%,29.7%±3.3%,47.9%±3.5% respectively,P<0.05),and the production of reactive oxygen species(ROS)also increased in time-dependent manner(14.0%±0.5%,36.1%±3.0%,38.2%±6.0%,48.3%±12.4%,P<0.05).The HepG2 cells showed typical morphologic change of endoplasmic retieulum stress induced by 2 μmol/L TG after 36 h and 48 h.DNA ladder was observed at the same concentration and time point correspondingly.Endoplasmic reticulum stress mediated-apoptosis was confirmed by Western blot.Both 10 mmol/L and 20 mmol/L NAC could protect ceils from apoptosis.The ratio of apoptotic cells decreased to 14.0%±1.3% and 11.0%±0.3%,respectively.The production of ROS decreased to 34.7%±0.8% and 31.5%±2.9%,respectively.The effect was related to the concentration of NAC.Conclusions As a Ca2+-adenosine triphoshatase inhibitor,TG may disrupt intracellular calcium homeostasis,which can induce endoplasmie reticulum stress and apoptosis.NAC,the precursor of the synthesis of-SH,can directly inhibit the ROS reaction and alleviate liver damage,which may play a role in the treatment of liver failure.

0.05).2.There were significant differences in the levels of TNF-? after treated for 4 weeks(P0.05).4.There were no significances between the 2 groups before and after treated 6 weeks in the change of liver.Conclusions Matrine injection can inhibit HCMV DNA replication.It can also control the expression of TNF-? and regulate the function of the immune system.There fore matrine injection has an antivirus efficacy in HCMV infection.

Objective: To observe the clinical efficacy of spleen-invigorating and qi-benefiting pediatric massage (tuina) for treating recurrent respiratory tract infection in children with cerebral palsy due to qi deficiency of spleen and lung. Methods: A total of 70 children with cerebral palsy who suffered from recurrent respiratory tract infection due to qi deficiency of spleen and lung were randomized into an observation group and a control group by the random number table method, with 35 cases in each group. Both groups were treated with conventional rehabilitation training, while the observation group was given additional spleen-invigorating and qi-benefiting pediatric massage, and the control group additionally took oral Yu Ping Feng granule. The clinical efficacy was evaluated by the total effective rate, traditional Chinese medicine (TCM) symptom score, and serum levels of immunoglobulin (Ig) A, IgM and IgG. Results: The difference in total effective rate between the two groups after treatment was statistically significant (P<0.05). After treatment, the TCM symptom scores in both groups decreased to varying degrees than those before treatment, and the intra-group differences were statistically significant (all P<0.01); the differences in the scores of various TCM symptoms between the two groups were statistically significant (all P<0.05). After treatment, the levels of IgA, IgM and IgG of the children in both groups increased to varying degrees, and the intra-group differences were statistically significant (all P<0.05). The between-group differences in the IgA, IgM and IgG levels after treatment were statistically significant (all P<0.05). Conclusion: Spleen-invigorating and qi-benefiting pediatric massage can effectively treat recurrent respiratory tract infection due to qi deficiency of spleen and lung in children with cerebral palsy, relieve the clinical symptoms and improve immune function, and thus is worthy of clinical promotion and application.

It was hypothesized that the VPAC1 agonist may exert anti-obesity functions because VPAC1 is involved in the anorexigenic effects and the anti-inflammatory function of pituitary adenylate cyclase-activating polypeptide (PACAP)/vasoactive intestinal polypeptide (VIP). Furthermore, our in vitro test showed that the expression of VPAC1 increased significantly after the 3T3-L1 adipocytes were differentiated, and that incubation of adipocytes with VPAC1 agonist (10-1 000 nmol/L per 1x10(6) cells) resulted in stimulation of lipolysis. To test the effect of VPAC1 agonist [Lys15, Arg16, Leu27]-VIP (1-7) GRF (8-27) on diet-induced obesity (DIO), we further designed the following two in vivo experiments: (1) Mice were fed on high-fat diet (HFD) and intraperitoneally (i.p.) treated with VPAC1 agonist simultaneously for 28 d; (2) Mice were given HFD for 35 d, and subsequently fed on the same HFD and i.p. treated with VPAC1 agonist for the next 28 d. The physiological indices, including body weight, weight of white adipose tissue, plasma glucose and blood lipid, were collected. The results showed that treatment with VPAC1 agonist inhibited ingestion significantly and prevented the elevations in body weight and the weights of the white adipose tissues (epididymal and dorsal) induced by HFD. The increases in plasma glucose, cholesterol, triglycerides and LDL induced by HFD were also down-regulated in mice treated with VPAC1 agonist. VPAC1 agonist treatment also improved the glucose tolerance. Therefore, VPAC1 agonist treatment inhibits the development of the obesity induced by HFD and helps to improve the morbidities associated with DIO.
3T3-L1 Cells ; Adipocytes ; drug effects ; Animals ; Body Weight ; Diet, High-Fat ; Mice ; Obesity ; drug therapy ; Receptors, Vasoactive Intestinal Polypeptide, Type I ; agonists ; Vasoactive Intestinal Peptide ; pharmacology

OBJECTIVETo investigate the dynamic changes of neurofilament contents in rat's spinal cord induced by 2, 5-hexanedione (2, 5-HD), and explore the molecular mechanism of n-hexane neuropathy.

METHODSMale Wistar rats were administered at a dosage of 400 mg/kg/day 2, 5-HD for 2, 4 and 8 weeks respectively. HD-induced neurological defects were detected and quantified using gait score, and the relative lev-els of NF-H, NF-M, and NF-L in spinal cords of rats were determined by Western Blotting.

RESULTSExposure to 2, 5-HD produced progressive gait abnormalities, which suggested that the rat model of 2, 5-HD-induced neurotoxicity was established successfully. Western-Blotting results showed that NFs content in spinal cord demonstrated a progressive decline as the intoxication continued. In the supernatant fraction, compared to the controls, NF-H con-tent decreased by 15.7%, 57.0%, and 58.0% respectively after 2, 4, and 8-week treatment with 2, 5-HD (P < 0.01); accordingly, NF-M decreased by 36.0%, 61.3%, and 65.2% respectively (P < 0.01); NF-L decreased by 20.8%, 43.9%, and 44.3% respectively (P < 0.01). In the pellet fraction, the contents of NF-H in groups of 4 and 8 weeks' exposure to HD decreased by 35.6% and 43.2%, respectively (P < 0.01), and those of NF-L decreased by 26.4% and 42.1%, respectively (P < 0.01) when compared to the control. Further-more, NF-M contents in groups of 2, 4 and 8 weeks' exposure decreased by 23.3%, 33.9%, and 63.7% respectively (P < 0.01). The NFs level in spinal cords was highly correlated with gait abnormality of treated rats as the intoxication went on. Multiple correlation coefficients of NF-H, NF-M, and NF-L content with gait score of HD-treated rat were 0.8912, 0.9282 and 0.8981 (P < 0.01) respectively.

CONCLUSIONThe declines of NFs are high-ly related to neurobehavioral abnormality of 2, 5-HD-treated animals, and involved in the development of n-hexane neuropathy.

Animals ; Disease Models, Animal ; Gait ; drug effects ; Hexanones ; toxicity ; Male ; Neurofilament Proteins ; metabolism ; Rats ; Rats, Wistar ; Spinal Cord ; drug effects ; metabolism