Objective To investigate the clinic significance of real-time fluorescent quantity polymerase chain reaction(RT-PCR) for detection of Mycoplasma pneumoniae(MP) DNA in patients suspected with MP infection.Methods A total of 1 402 samples,including serum,sputum,pleural fluid,nasopharyngeal swab,alveolar irrigating solution and bronchial irrigating solution,were detected for MP-DNA by using RT-PCR.Results The total positive rate all samples were 12.20%.The positive rate of serum was the lowest,which was 2.36%.The positive rates of sputum,alveolar irrigating solution and bronchial irrigating solutions were relatively high,which were 62.96%,77.08% and 88.71%,respectively.Conclusion RT-PCR could be fitted for the detection of MP-DAN in various samples,which could be effective method for the diagnosis of MP infection.

Objective:To investigate the effects of Porhyromonas endodontalis(P.e)liopolysaccharide(LPS)on the expression of IL-23mRNA and protein in mouse osteoblasts MC3T3-E1 and the role of phosphatidylinositol 3-Kinases (PI3K)signaling pathway in this process.Methods:MC3T3-E1 cells were treated with different concentrations of P.e LPS for different hours,or pretreated with LY294002,a special PI3K inhibitor.The IL-23 mRNA and protein expression levels were detected by real-time PCR and Western blot respectively.Results:The level of IL-23 mRNA increased in MC3T3-E1 cells with the increase of concentration and the treatment time of P.e LPS(P <0.05).The IL-23 protein expression was increased by P.e LPS in a time dependent manner(P <0.05).The mRNA and protein of IL-23 decreased(P <0.05)after pretreatment with LY294002.Conclusion:P.e LPS can induce the expression of IL-23 mRNA and protein in MC3T3-E1 cells,and the PI3K signaling pathway may play a part in this process.

Objective To investigate the correlation of multidrug resistance gene 1 (MDR1),NR3C1 gene polymorphisms and clinical risk factors with efficacy,dependence,and resistance of glucocorticoid (GC) in patients with inflammatory bowel disease (IBD).Methods Anti coagulation blood samples of 196 healthy controls and 105 IBD patients received GC therapy were collected.There were 62 ulcerative colitis (UC) and 43 Crohn's disease (CD) in the IBD patients.The number of GC sensitive,GC dependent and GC resistant of UC patients were 36,13 and 13,respectively,and those of CD patients were 24,11 and eight.GC refractoriness included GC dependence and resistance.The genotype of MDR1 C3435T and NR3C1 Bcl Ⅰ of all the subjects was detected by the restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR).The correlation between each genotype frequency,clinical features of patients with IBD and the efficacy of GC treatment was analyzed by Chisquare test,Fisher exact probability method or t test.Results Among UC patients,the disease course of GC refractory group and GC resistant group was longer than that of GC sensitive group ((6.660±1.523)years,(6.500±1.111) yearsvs (3.350±0.697) years,t=2.211,P=0.031; t=2.930,P=0.005).The serum level of C reaction protein (CRP) of GC refractory group was higher than that of GC sensitive group ((47.628±13.913) mg/Lvs (16.854±4.121) mg/L,t=2.121,P=0.047).The chronic relapse type was more common in GC refractory UC patients (Fisher exact probability method,P=0.035),and severe patients were more common in UC with GC resistance (Fisher exact probability method,P=0.021).The white blood cell count of GC resistant and GC refractory CD patient was lower than that of GC sensitive CD patients ((5.710 ± 0.604) ×109/L,(5.878±0.405) × 109/L vs (7.814 ±0.670) × 109/L,t=2.334,P=0.028; t=2.045,P=0.018).Patients with extraqntestinal manifestations was more common in CD with GC resistance (Fisher exact probability method,P=0.035).There was no statistically significant difference in the frequencies of MDR1 C3435T,NR3C1 Bcl Ⅰ genotypes,allelic genes and gene carrier among control group and GC sensitive dependent and resistant group of IBD patients.However,the frequency of MDR1 C3435T gene carrier was significantly different between GC sensitive group and GC refractory group,especially between GC sensitive group and GC resistance group (68.33％ vs 48.89％,x2 =4.051,P=0.044; 68.33％ vs 42.86％,x2 =4.274,P =0.039).Conclusions GC sensitivity of IBD patients with MDR1 C3435T loci T gene carrier was higher than that of IBD patients without T gene carrier.NR3C1 gene polymorphisms was not related with GC resistance and GC dependence.Compared with GC sensitive IBD patients,in GC resistant and GC dependent IBD pantient UC patients with long disease course,chronic relapse type,severe type,high level of CRP and CD patients with low white blood cell count and extra-intestinal manifestations were more common.

PurposeTo investigate the feasibility of using low concentration contrast agent and low tube voltage in the light and moderate weight's abdominal contrast-enhanced CT scan, in order to find an optimal solution to reduce radiation dose and iodine intake.Materials and Methods Forty patients with light weight whose body mass indexes (BMI) were lower than 20 kg/m2 were randomly divided into group A1 (n=20) and group B1 (n=20). Meanwhile, another 40 patients with moderate weight whose BMI ranged from 20 kg/m2 to 25 kg/m2 were randomly divided into group A2 (n=20) and group B2 (n=20). Low concentration contrast agent and low tube voltage (Visipaque 270 mgI/ml, 100 kV) were used in both group A1 and group A2 in abdominal enhanced CT scan. While both group B1 and group B2 used conventional scan solution (Omnipaque 300 mgI/ml, 120 kV) in abdominal enhanced CT scan. Then the contrast noise ratio (CNR), the image quality score and the effective radiation dose (ED) were compared among the four groups.Results The CNR and image quality score at artery phase and portal phase were neither significantly different between group A1 and group B1, nor between group A2 and group B2 (t=-1.539-0.000,P>0.05). The CNR and image quality score of the liver at artery phase in group B1 were signiifcantly higher than those in group A2 and group B2 (P<0.05).Conclusion The solution of using low concentration contrast agent and low tube voltage in contrast enhanced scan can achieve the same high quality abdominal image with reduced iodine intake and radiation, compared with the application of conventional enhanced scan; BMI has rather great impact on image quality score at arterial phase and little impact on that at portal phase. So it is suggested that the protocol of liver contrast-enhanced CT scan may choose reduction of voltage at portal phase so as to reduce radiation.

Objective:To investigate the extracellular signal-regulated kinase 5 (ERK5) and matrix metallo proteinase-9 (MMP-9) expres-sion levels in osteosarcoma tissues and their clinical significance. Methods:The ERK5 and MMP-9 expression levels in 71 specimens of osteosarcoma tissue and 40 specimens of normal bone tissue were detected by immunohistochemistry. The relationship between ERK5 and MMP-9 expression levels, their clinical characteristics, and prognosis of patients with osteosarcoma were analyzed. Results:The positive expression of ERK5 and MMP-9 in osteosarcoma tissues was 85.9%(61/71) and 74.65%(53/71), respectively, which were significantly higher than those in normal bone tissues at 12.5%(5/40) and 10.0%(4/40) (all P<0.05). The positive expression of ERK5 and MMP-9 was associated with Enneking stage and metastasis (all P<0.05). Kaplan-Meier analysis showed that the survival duration of patients with positive ERK5 and MMP-9 expression levels was shorter than those of the patients in the negative expression groups (all P<0.05). Univariate analysis of COX proportional hazards regression model revealed that tumor size, Enneking stage, metastasis, and positive ERK5 and MMP-9 expression levels are relevant to the overall survival of patients with osteosarcoma (all P<0.05). Multi-variate analysis of COX proportional hazards regression model confirmed that Enneking stage, metastasis, and positive ERK5 and MMP-9 expression levels can act as independent prognostic factors for osteosarcoma patients (all P<0.05). Conclusion:The ERK5 and MMP-9 expression levels are high in osteosarcoma tissues and are related to the clinical characteristics and prognosis of patients with osteo-sarcoma. Thus, ERK5 and MMP-9 expression levels may play important roles in osteosarcoma development and progression.

Notch signaling is highly conserved in evolution and regarded as a key factor in cell fate determination. It mediates cell-to-cell interactions that are critical for embryonic development and tissue renewal, and is involved in the occurrence and metastasis of neoplasm. Recent researches have found that such signaling plays an important role in modulating the differentiation of chondrocytes, osteoblasts and osteoclasts. Dysfunction of Notch signaling can result in many skeletal diseases such as bone tumor, disorders of bone development or bone metabolism.
Animals ; Bone Development ; Bone Diseases ; genetics ; metabolism ; Bone and Bones ; metabolism ; Humans ; Osteoblasts ; cytology ; metabolism ; Osteogenesis ; Receptors, Notch ; genetics ; metabolism ; Signal Transduction

OBJECTIVETo investigate the effects of advanced oxidation protein products (AOPP) on epithelial-to-mesenchymal transition (EMT) in cultured human proximal tubular epithelial cells (HK-2) and explore the mechanism.

METHODSHK-2 cells treated with 50, 100, 200, and 400 µg/ml AOPP or 50 µg/m bovine serum albumin (BSA) for 24 h, or with 200 µg/ml AOPP for 0.5, 1, 3, 6, 12, and 24 h were examined for the protein expression of α-SMA and E-cadherin. In cells pretreated with diphenyleneiodonium (DPI) or cytoplasmic superoxide dismutase (C-SOD), the effects of 50 µg/ml BSA and 200 µg/ml AOPP were assessed on the expressions of α-SMA and E-cadherin, malondialdehyde (MDA) level, superoxide dismutase (SOD) activity, catalase (CAT) activity, and glutathione peroxidase (GSH-px) activity.

RESULTSAOPP treatment up-regulated α-SMA expression and down-regulated E-cadherin expression in a dose- and time-dependent fashion. AOPP exposure of the cells resulted in increased MDA level and lowered activities of SOD, CAT and GSH-PX. DPI and C-SOD partially attenuated the effects of AOPP on α-SMA, E-cadherin, MDA, SOD, CAT and GSH-px.

CONCLUSIONAOPP can induce EMT in cultured HK-2 cells via oxidative stress, and this effect can be attenuated by inhibiting the activation of NADPH oxidase and using antioxidants to delay the progression of renal interstitial fibrosis.

Actins ; metabolism ; Advanced Oxidation Protein Products ; Antioxidants ; metabolism ; Cadherins ; metabolism ; Catalase ; metabolism ; Cell Line ; Cells, Cultured ; Down-Regulation ; Epithelial Cells ; cytology ; Epithelial-Mesenchymal Transition ; Glutathione Peroxidase ; metabolism ; Humans ; Malondialdehyde ; metabolism ; NADPH Oxidases ; metabolism ; Oxidative Stress ; Superoxide Dismutase ; metabolism ; Up-Regulation

OBJECTIVETo investigate the association between single nucleotide polymorphisms (SNPs) of the transmembrane protein 39A (TMEM39A) at the loci 1880G/A, 2442T/G, and 2456A/T and systemic lupus erythematosus (SLE) in Chinese Han patients.

METHODSTMEM39A gene polymorphisms at 3 loci (1880G/A, 2442T/G, 2456 A/T) were analyzed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 110 Chinese Han patients with SLE and 80 normal control subjects, and the allele and genotype frequencies were compared by Chi-square test between the two groups.

RESULTSBoth the genotype frequencies (AA, GA and GG) and allele frequencies (A and G) at 1880G/A differed significantly between SLE cases and the normal controls (P=0.002 and P=0.044, respectively). The two groups also showed significant differences in the genotype frequencies (GG, TG and TT) (P=0.001) and allele frequencies (G and T) (P=0.041) at 2442T/G. No significant differences were found in the genotype frequencies (TT, AT and AA) or allele frequencies (T and A) at 2456A/T between the two groups (P>0.05). The allele and genotype frequencies of the 3 SNPs showed no significant differences between lupus nephritis (LN) patients and non-LN patients.

CONCLUSIONThe TMEM39A polymorphisms at 1880G/A and 2442T/G, but not at 2456 A/T gene, may be associated with the susceptibility to SLE in Chinese Han population. The genotype or allele frequencies of the 3 SNPs have no effect on the incidence of lupus nephritis.

Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Female ; Gene Frequency ; Genotype ; Humans ; Lupus Erythematosus, Systemic ; genetics ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Polymorphism, Single Nucleotide ; Young Adult

Objective To investigate the association between single nucleotide polymorphisms (SNPs) of the transmembrane protein 39A (TMEM39A) at the loci 1880G/A, 2442T/G, and 2456A/T and systemic lupus erythematosus (SLE) in Chinese Han patients. Methods TMEM39A gene polymorphisms at 3 loci (1880G/A, 2442T/G, 2456 A/T) were analyzed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 110 Chinese Han patients with SLE and 80 normal control subjects, and the allele and genotype frequencies were compared by Chi-square test between the two groups. Results Both the genotype frequencies (AA, GA and GG) and allele frequencies (A and G) at 1880G/A differed significantly between SLE cases and the normal controls (P=0.002 and P=0.044, respectively). The two groups also showed significant differences in the genotype frequencies (GG, TG and TT) (P=0.001) and allele frequencies (G and T) (P=0.041) at 2442T/G. No significant differences were found in the genotype frequencies (TT, AT and AA) or allele frequencies (T and A) at 2456A/T between the two groups (P>0.05). The allele and genotype frequencies of the 3 SNPs showed no significant differences between lupus nephritis (LN) patients and non-LN patients. Conclusion The TMEM39A polymorphisms at 1880G/A and 2442T/G, but not at 2456 A/T gene, may be associated with the susceptibility to SLE in Chinese Han population. The genotype or allele frequencies of the 3 SNPs have no effect on the incidence of lupus nephritis.

Objective To investigate the effects of advanced oxidation protein products (AOPP) on epithelial-to-mesenchymal transition (EMT) in cultured human proximal tubular epithelial cells (HK-2) and explore the mechanism. Methods HK-2 cells treated with 50, 100, 200, and 400μg/ml AOPP or 50μg/m bovine serum albumin (BSA) for 24 h, or with 200μg/ml AOPP for 0.5, 1, 3, 6, 12, and 24 h were examined for the protein expression of α-SMA and E-cadherin. In cells pretreated with diphenyleneiodonium (DPI) or cytoplasmic superoxide dismutase (C-SOD), the effects of 50μg/ml BSA and 200μg/ml AOPP were assessed on the expressions of α-SMA and E-cadherin, malondialdehyde (MDA) level, superoxide dismutase (SOD) activity, catalase (CAT) activity, and glutathione peroxidase (GSH-px) activity. Results AOPP treatment up-regulatedα-SMA expression and down-regulated E-cadherin expression in a dose- and time-dependent fashion. AOPP exposure of the cells resulted in increased MDA level and lowered activities of SOD, CAT and GSH-PX. DPI and C-SOD partially attenuated the effects of AOPP onα-SMA, E-cadherin, MDA, SOD, CAT and GSH-px. Conclusion AOPP can induce EMT in cultured HK-2 cells via oxidative stress, and this effect can be attenuated by inhibiting the activation of NADPH oxidase and using antioxidants to delay the progression of renal interstitial fibrosis.