90 extracted single rooted human teeth were inoculated with E.faecalis.The cannals of the teeth were irrigated with MTAD, 5.25% NaClO,compound chlorhexidine gargle,tinidazole mouthwash,distilled H2 O(infection control)and autoclave followed by distilled H2 O(autoclaving control),respectively(n =1 5).Then E.faecalis infection of the samples were examined.The results showed that MTAD had significantly greater antibacterial effects than 5.25% NaClO,tinidazole mouthwashes,compound chlorhexidine gargle and tinidazole mouthwash(P <0.05).There was no significant difference between each 2 of the other 3 irrigants.

The health care alliance in county was the vertical integration of medical service system,which was conductive to improve the overall efficiency of the county medical service,achieve tiered health care system,optimize regional health resources and enhance the capacity of primary health care services.It took Tianchang County,Anhui as a case study.It investigated and analyzed how the local took measures to carry out the health care alliance reform in organization structure,interest distribution,health human resources construction,information construction,division and coordination mechanism,medical service pattern,etc.,so as to provide references for implementing county medical alliance reform in other regions.

Objective To detect the expression of Ezrin in giant-cell tumor of bone,and to investigate its cilincal significance. Methods 60 cases of biopsy which had been confirmed as bone giant-cell tumors in our hospital from January 2008 to December 2013 were set as observation group;tumor tissues from 8 cases of reactive new bone in nonmalignant bone diseases,12 cases of osteoid osteoma and 11 cases of osteoblastoma in the corresponding period were set as control group. Protein and gene levels of Ezrin were tested with Western blotting method and real-time PCR detection,simultaneously proceeded the corresponding analysis combined with the clinical data of patients;60 cases of bone giant-cell tumor patients accepted tumor resection and pros-thesis replacement,2 courses of preoperative chemotherapy;mitochondria morphological changes of tumor tissue and Ezrin protein and genetic changes were observed before and after chemotherapy. Results In the giant-cell tumors of bone,the Ezrin protein mainly located in the cytoplasm,and its expression positive rate was much higher than that in reactive new bone of nonmalignant bone diseases(19. 7% ),osteoid osteoma(21. 2% )and osteoblastoma(20. 7% );the difference was statistically significant(χ2 = 4. 18,P = 0. 024),but no statistical difference in the Ezrin expression among the groups of osteosarcoma,osteoid osteoma and osteblastoma(χ2 =6. 18,P = 0. 087). In the giant-cell tumors of bone tissue after chemotherapy,mitochondria pyknosis and the phenomenon of liquid cavitation was less than that before the treatment,and Ezrin protein expression decreased and gene levels reduced[(23. 99 ± 1. 49)vs(20. 11 ± 1. 11),t = 5. 03,P = 0. 018)]. Conclusion The expression of Ezrin in giant-cell tumor of bone is much higher than other benign bone tumor,and it could be a biological marker for differentiating benign and malignant bone tumor. Early intervention in Ezrin may be helpful for reatment of giant-cell tumor of bone.

Objective To investigate the clinical features and treatment points of diabetes patients compli-cated with parapharyngeal abscess .Methods The clinical data of 12 diabetes mellitus patients complicated with parapharyngeal abscess were analyzed .Results 12 patients were all cured and discharged without concurrent medias-tinal abscess ,intracranial infection and serious complications such as bleeding .Conclusion The development of dia-betes mellitus complicated with parapharyngeal abscess is rapid ,we must confirm the diagnosis ,timely implementation of abscess incision and drainage ,effective drainage of pus ,keep the airway open ,actively control the infection at the same time strictly control the fluctuations in blood sugar ,strengthen supportive therapy ,thus to improve treatment and reduce the incidence of complications .

AIM: To investigate the pharmacodynamic effects of Huahong Capsule on the anti-bacteria,anti-inflammation,antipyretic and analgesic effects. METHODS: The death rate of mice infected by staphylococcus aureus or Escherichia coli was observed to investigate the antibacterial effects.Crotonoil-induced auricle edema,cotton ball-induced granuloma and trauma-induced toe congestion were performed to investigate the anti-inflammatory effects in mice.Endotoxin-induced fever was performed to observe the antipyretic effect in rabbit.Mouse twisting numbers induced by acetic acid,mouse pain threshold after heat stimulation and rat twisting numbers of dysmenorrhea were observed to investigate the analgesic effects. RESULTS: Huahong Capsule had a tendency to decrease the death rate of mice infected by bacteria.It could remarkably inhibit edema of auricle of ear,formation of granuloma and toe congestion in mice.It decreased obviously the temperature raised by endotoxin in rabbit.It could reduce mouse twisting numbers and prolong pain thresholds. CONCLUSION: Huahong Capsule has pharmacological effects in accordance with its clinical functions and uses.

AIM:To investigate the effect of cyclopamine on Hedgehog (HH) signaling, phenotypic transfor-mation and matrix accumulation induced by aristolochic acid (AA) in renal tubular epithelial cell NRK-52E.METHODS:NRK-52E cells were randomly divided into control group (treated with solvent only), AA group (treated with AA at con-centrations of 1, 5, 10 mg/L) and cyclopamine group (treated with AA at concentration of 10 mg/L plus cyclopamine at concentrations of 1, 5, 10μmol/L).After cultured for 24 h, the mRNA expression of Ptch1, Smo,α-SMA, E-cadherin, ZO-1, BMP-7, type I collagen and type III collagen was quantified by real-time PCR.The protein levels of Shh and TGF-β1 were detected by ELISA .Immunofluorescence staining was used to evaluate the expression of Ptch 1, Smo,α-SMA, E-cadherin and type III collagen in the NRK-52E cells.RESULTS: AA increased the expression of TGF-β1, α-SMA and type III collagen, decreased the expression of E-cadherin and ZO-1 protein, and down-regulated the expression of Ptch1, Shh and Smo mRNA in the NRK-52E cells, indicating that AA activated HH signaling , and phenotypic transformation and matrix accumulation occurred in AA-treated NRK-52E cells.Treatment with cyclopamine inhibited HH signaling by decrea-sing Smo expression and increasing Ptch 1 expression.Moreover, cyclopamine also down-regulated the expression of TGF-β1,α-SMA, type I collagen and III collagen , and up-regulated the expression of BMP-7, ZO-1 and E-cadherin.CON-CLUSION:AA induces phenotypic transformation and matrix accumulation in renal tubular epithelial cells , which can be inhibited by cyclopamine treatment .The possible mechanism is that cyclopamine suppresses the activation of HH signaling , resulting in the reduction of epithelial-to-mesenchymal transition and matrix deposition .

Objective To investigate the expression of monocyte-macrophage-related factors and interstitial fibrosis in kidney tissues of rats with ureter obstruction and recanalization .Methods Forty-eight male Spragur-Dawley rats were divided randomly into the obstructive group:sham (n=6), unilateral ureteral obstruction(UUO)3 days (n=6), UUO 7 days (n=6), and UUO 14 days (n=6) and recanalization group:bilateral ureteral obstruction(RBUO)0 day (n=6), 3 days after RBUO (n=6), 7 days after RBUO (n=6), and 14 days after RBUO (n=6).The kidneys were excised on day 3, 7, and 14, and the deposition of collagen fibers in kidney was detected with HE and Masson staining . Immunohistochemical analysis was performed to evaluate the protein expressions of monocyte chemoattractant protein -1 (MCP-1), macrophage colony-stimulating factor (M-CSF) and activated-macrophage marker CD68.Real-time PCR was used to detect the mRNA expressions of MCP-1 and M-CSF.TGF-β1 levels were determined by ELISA .Results Fibrosis observed with HE and Masson staining was obviously increased in kidney tissue of UUO rats , and aggravated as time prolonged, but alleviated in rats with recanalization .TGF-β1 levels were increased obviously in the UUO group , but decreased in rats with recanalization compared with those in BUO rats .In UUO rats, mRNA and protein expression levels of MCP-1 and M-CSF were increased .MCP-1 and M-CSF expression was gradually decreased in rats with recanalization compared with those in BUO rats .The dynamic change in expression of MCP-1 and M-CSF in both UUO rats and recanalization rats was consistent with the change in expression of CD 68. Conclusion Dynamic change in expression of MCP-1 and M-CSF in kidney tissues reflects change of activated and accumulated monocyte -macrophages , which may be one of the major mechanisms contributing to fibrosis induced by ureter obstruction .Renal fibrosis is alleviated by down-regulated expression of monocyte-macrophages factors with recanalization operation .

Objective To investigate the effects of IL-12 coexpression level on antigen expression and immune responses induced by HBsAg DNA vaccination. Methods DNA vaccine plasmid pHBV carrying codon-optimized preS2-S gene of reference sequence CHN-HBV07-C in China was constructed. Three DNA vaccine plasmids pHBV-12i, pHBV-12l and pHBV-12h were also constructed by subcloning three different IL-12 expression cassettes with various expression strengths to plasmid pHBV, respectively. Expression levels of IL-12 and HBsAg in vaccine plasmid-tranfected 293T cells were measured by quantitative ELISA. DNA vaccines were administered intramuscularly to BALB/c mice and HBsAg-specific cellular immune responses were determined by IFN-γ ELISPOT. HBsAg-specific antibodies were tested by Chemiluminescence Quantitative Immunoassay. Results The HBsAg expression level in 293T cells was 70 ng/ml when transfected by plasmid pHBV without IL-12 expression cassette, and the HBsAg level was 18 ng/ml when transfected by plasmid pHBV-12l carrying low-level IL-12 expression cassette, whereas the HBsAg level was only 6 ng/ml when transfected by plasmid pHBV-12h carrying high-level IL-12 expression cassette.Results of DNA vaccination revealed that HBsAg-specific humoral and cellular immune responses were significantly decreased in mice administering vaccine pHBV-12h carrying high-level IL-12 expression cassette. Although HBsAg-specific antibody responses in mice inoculated with pHBV-12l were also decreased when compared with those in pHBV-vaccinated mice without IL-12 expression, the HBsAg-specific cellular immune responses were significantly increased. Conclusion High-level coexpression of IL-12 may suppress the expression of HBsAg, Whereas modest coexpression of IL-12 significantly enhanced the HBsAg-specific T cell responses induced by DNA vaccination. Therefore, it is so important to balance the expression between adjuvant and antigen to enhance the immune response.

Objective To detect mutations of the ABCA12 gene in 2 Chinese families with autosomal recessive congenital ichthyosis (ARCI).Methods According to the typical clinical manifestations,two probands were diagnosed with ARCI.DNA was extracted from the peripheral blood samples collected from the patients and their parents.High-throughput sequencing was conducted by using multi-gene array for genetic skin disorders to determine mutation sites in the probands,and then DNA isolated from the probands and their parents were bidirectionally verified by Sanger sequencing.Results Two compound heterozygous mutations (c.2759A＞G and c.7004A＞G) in the ABCA12 gene were found in the proband 1,and another two compound heterozygous mutations (c.6163_6164insT and c.7406G＞A) were identified in the proband 2.The parents of the two probands were heterozygous carriers of one of the two mutations in the ABCA12 gene.Function prediction for the 4 mutations showed that all of the 3 missense mutations (c.2759A＞G,c.7004A＞G and c.7406G＞A) may exert pathogenic effect,and fragnin encoded by the frameshift mutation c.6163_6164insT may also affect protein function,c.2759A＞G and c.6163_6164insT were newly identified mutation sites.Conclusion The compound heterozygous mutations in the ABCA 12 gene are the causative mutations responsible for ARCI in the two probands of the two pedigrees.