Objective: To construct mcpr1prokaryoti c expression vector and to express MCPR1 protein.Methods:PCR was used to obtain coding region of mcpr1. Construction of a high-level fusion protein expression vector pGEX-4T-mcpr1 was conducted by inserting the fra gment of coding region of mcpr1into a fusion protein expression vector pGEX -4T-1. Then the recombinant plasmid was transferred into E. colito prepar e the MCPR1/GST fusion protein. DNA sequencing and endonucleases digesting were used to check the coding region. Results:pGEX-4T- mcpr1 wa s constructed successfully and the coding region was inserted into the vector co rrectly. A new protein band of 36 000 was observed by SDS-PAGE analysis after i nduction by IPTG. The 36 000 protein amounted to 39 percent of the total prote in and existed mostly in precipitation of broken bacteria. Conclusion: MCPR1 protein can be expressed in E. coliexpression system and purif ied initially.

Objective To investigate the value of combinedevaluation of serum lipoprotein associated phospholipase A2 (LP-PLA2) and cystatin C (CysC) on systemic lupus erythematosus (SLE) patients with lupus erythematosus.Methods A total of 177 patients with SLE were enrolled in the General Hospital of People's Liberation Army from June 2000 to February 2017,87 cases in lupus group (SLE group),90 cases in lupus nephritis group (LN group),60 cases in healthy control group.The concentration of LP-PLA2 and CysC were measured.The ROC curves was established for estimation of the cutoff values of two indexes in SLE patients with LN,and the diagnostic efficacy of different diagnostic criteria was compared.Results The LP-PLA2 and CysC concentration in the LN group were higher than those in the SLE group (Z=-2.512,-5.688,all P＜0.05).The cutoff value of LP-PLA2 was 245.5 U/L and the cutoff value of CysC was 1.235 mg/L on the risk assessment of in SLE patients with LN.When using the parallel method for combined LP-PLA2 and CysC to evaluate the risk of LN,the sensitivity and negative predictive value would be significantly improved (x2 =9.461,4.377,all P＜ 0.05),the efficiency was better.Conclusion Parallel assessment of serum LP-PLA2 and CysC have clinical value on assessment of the risk of LN in SLE patients.

Objective:To evaluate the physiological roles of bmp3,bmp4 and bmp7 during the formation of permanent tooth roots in dog. Methods:The expression of bmp3,bmp4 and bmp7 mRNA at different stages of the development of permanent tooth roots was examined by in situ hybridization in 3 dogs aged 12-18 weeks. Results:bmp3 was found in dental sac surrounding the germs at the early stage of tooth root development, and in cementoblasts and periodontal cells at the later stage. bmp4 was found in odontoblasts, dental papilla and osteoblasts. bmp7 positive signals was found only in epithelial cells of root sheath around cervical circulus at early stage, then located in cementoblasts and odontoblasts at later stage. Conclusion:The spatiotemporal expressions of bmp3,bmp4 and bmp7 are widely diverse, indicating that they participate in the regulation of tooth root development.

OBJECTIVE To investigate the distribution of Pseudomonas aeruginosa and its antibiotic resistance in(emergency) intensive care unit(EICU) patients with lower respiratory infections.METHODS The data of the pathogens isolated in sputum from the 25 patients with lower respiratory infections admitted to EICU from(Aug) 2005 to Feb 2006 were collected and analyzed.RESULTS Eighty-one bacteria strains were found in these 25 EICU cases.The Gram-negative bacteria were the main pathogens of infection(43.2%),after them were Gram-positive bacteria(32.1%),and fungi(24.7%).The percentage of P.aeruginosa infection was 13.6% of all(patients).The antibiotic resistance for P.aeruginosa was found to the antibiotics,such as cefotaxime,gentamicin,(SMZ-TMP),minocycline,levofloxacin,and gatifloxacin.CONCLUSIONS The P.aeruginosa infection and its(antibiotic) resistance should be paid more attention in the treatment of the lower respiratory infection patients in EICU.

Humans ; Infant, Newborn ; Lower Extremity ; pathology ; Lymphedema ; congenital ; Male

Abscess ; etiology ; Adult ; Humans ; Male ; Nasal Septum ; pathology ; Tooth, Impacted ; complications

OBJECTIVEThis study aims to detect microRNA-17(mir-17) expression on the osteogenic differentiation of advanced glycation end products (AGEs)-stimulated hunman periodontal ligament stem cells (HPDLSCs) and to analyze the influence of these cells on this process.

METHODSHPDLSCs were isolated using limited dilution technique. After osteogenic differentiation occurred, different time points of mir-17 expression in the experimental groups were detected by real time polymerase chain reaction (PCR). The mir-17 overexpression and inhibition were evaluated using cell transfection technique. Differences in gene expressions were detected by real time PCR; differences in protein expressions were analyzed by Western blot.

RESULTSThe mir-17 expression was reduced after osteogenic differentiation occurred at 3, 7, and 14 d compared with that in the control group (P < 0.05). The expression levels of bone sialoprotein (BSP), Runt-related transcription factor-2 (Runx-2)and alkaline phosphatase (ALP) in the experimental groups were lower than those in the mimic control group when mir-17 expression increased. In addition, the protein expression levels of Runx-2 in the experimental groups were lower than those in the control group. The expression levels of BSP, Runx-2 and ALP in the experimental groups were higher than those in the inhibitor control group when mir-17 expression decreased. Likewise, the protein expression levels of Runx-2 in the experimental groups were higher than those in the control group.

CONCLUSIONAGEs inhibit the osteogenic differentiation of HPDLSCs by affecting mir-17 expression.

Alkaline Phosphatase ; Cell Differentiation ; Glycation End Products, Advanced ; Humans ; MicroRNAs ; Osteogenesis ; Periodontal Ligament ; Stem Cells

OBJECTIVEThe effect of advanced glycation end products (AGEs) on the osteogenic differentiation of humanperiodontal ligament stem cells(hPDLSCs) was discussed. Changes in the Wnt signaling pathway during glycation were also determined.

METHODSIn vitro tissue explanting method was primarily applied. Limiting diluted clone was cultured to obtain hPDLSCs in vitro. The subjects were divided into two groups: the healthy group (N-hPDLSCs) and the AGEs-stimulating group (A-hPDLSCs). Osteoblast mineralization was induced in the experimental groups. The following processes were performed: alizarin red staining; alkaline phosphatase (ALP) staining; real time polymerase chain reaction (real time PCR) for detecting osteogenic genes and Wnt classical pathway-related factors, DKK-1 and β-catenin; Western blot analysis. Bone protein and β-catenin were correlated in the nuclear expression.

RESULTSThe cells were osteogenically induced. ALP staining showed that the N-hPDLSCs displayed the deepest color. Alizarin red staining indicated that the A-hPDLSCs group had less calcified nodules than the N-hPDLSCs group. The real time PCR results suggested that the expression of relative osteogenic genes in A-hPDLSCs was quite low. Statistically significant differences in differentiation were found between groups (P < 0.05). The Western blot result was similar to that of real time PCR. Classical Wnt signaling pathway-related factor β-catenin was higher in A-hPDLSCs than in N-hPDLSCs. By contrast, DKK-1, which is an inhibitor in the Wnt pathway, had a significantly lower expression rate in A-hPDLSCs than in N-hPDLSCs. The Western blot result also showed that β-catenin expression in the nucleoprotein in A-hPDLSCs was notably higher than in N-hPDLSCs.

CONCLUSIONAGEs can inhibit hPDLSCs osteogenic differentiation. AGEs induce changes in the normal periodontal ligament stem cells classical Wnt pathway. Canonical Wnt pathway is reactivated because of AGEs stimulation.

Cell Differentiation ; Glycation End Products, Advanced ; Humans ; In Vitro Techniques ; Osteoblasts ; Osteogenesis ; Periodontal Ligament ; Stem Cells ; Wnt Proteins ; Wnt Signaling Pathway ; beta Catenin

Objective:To observe the temporal and spatial expression and function of mcpr1 gene during murine tooth germ development.Methods:The expression of MCPR1 at different stages of mouse tooth germ were detected by immunohistochemical staining.Results:MCPR1 expression was detected at all stages of tooth germ, but the distribution patterns at various stages were different. It indicated that the temporal and spatial expression pattern of MCPR1 during murine tooth germ development was specific.Conclusion:mcpr1 might play an important role in modulating the differentiation and mature of enamel organ.

Objective To explore the effect of reduced glutathione and venous systemic oxygen perfusion on apoptosis and ultrastructure of hepatocytes in rat steatotic liver grafts. Methods Before liver transplantation grade Ⅱ steatotic liver model was established by a diet consisting of 79% standard diet,20% lard and 1% cholesterol for 6 weeks. In pretreatment group, the donor received intraperitoneal injection of reduced glutathione at a dosage of 500 mg/kg/body weight 3 times a day for 2 days, and intrahepatic venous oxygen perfusion for 6 hours while kept in cold preservation. Results Preconditioning measures in steatotic liver grafts significantly decreased the hepatocytes necrosis (38?10)% vs (17?6)%, P