Objective To investigate the expression pattern of microRNA (miRNA) in the kidneys of unilateral ureteral obstruction (UUO) rats and to identify specific miRNA related to renal interstitial fibrosis (RIF).Methods Forty-eight male SD rats were divided into two groups:UUO group and sham-operated (Sham) group.Rats were sacrificed at 3,7 and 14 days after operation.Histologic changes were examined by Masson staining.Forty-eight selected miRNAs were examined by stem-loop real-time qPCR.Results At the 3rd day after operation,obstructed kidneys from operation rats showed mild edema in the interstitium and mononuclear cell infiltration.At the 7th day after operation,focal interstitial fibrosis was observed.At the 14th day after operation,fibrosis became more severe.The Sham kidneys showed no pathological changes.At the 3th day after operation,25 miRNAs were differentially expressed.At the 7th day after operation,24 miRNAs were aberrantly expressed,whereas 21 miRNAs were differentially expressed at the 14th day after operation (P＜0.05).Among these miRNAs,miR-132,miR-192,miR-194,miR-29c and miR-203 were consistently up-regulated or down-regulated in a time-dependent manner after operation.There were significantly correlations between the expression of five miRNAs and severity of tubulointerstitial injury (P＜0.05).Conclusions There are at least 20 miRNAs differentially expressed in the process of RIF induced by UUO.There are significantly correlations between the expression of miR-132,miR-192,miR-194,miR-29c and miR-203 and the severity of tubulointerstitial injury.They may be closely related to RIF.A further study is needed.

Objective To explore the relationship between HPV infection and polymorphisms of IL-1β and EGFR and lung cancer in non-smokers female. Methods A case-control study method was used, with 197 patients with lung cancer of non-smoking female lung cancer patients and 87 healthy female. The HPV DNA was detected with PCR technology, and the polymorphisms of IL-1β (-31C/T、-511T/C) and EGFR (-216G/T、-191C/A、8227G/A、D994D and R497K) were detected with TaqMan single nucleic acid polymorphism typing technology. Non-conditional Logistic regression was used to analyze the correlation between polymorphic genotype and lung cancer in non-smokers. Results The detection rate of HPV 16 in non-smoking female with lung cancer was significantly higher than that in the control group (P < 0.05). There was no significant difference in the gene polymorphisms of IL-1β-511 T/C and EGFR-191 C/A, -216 G/T, D99D, and R497K between the two groups (P > 0.05). There was statistically significant difference in IL-1β-31 C/T locus gene polymorphism between the two groups (P < 0.05), andthe risk of T/T genotype lung cancer was significantly increased (OR=0.25, 95% CI: 0.16～0.55). There was no statistically significant difference in the -191C/A, -216G/T, D99D and R497K gene polymorphisms of the EGFR gene between the two groups (P>0.05). there was statistically significant difference in EGFR-8227GA gene polymorphism between the two groups (P < 0.05), and the risk of G/A genotype lung cancer was significantly increased (OR=0.22, 95% CI: 0.18-0.57). Conclusions HPV 16 infection, IL-1β-31 T/T genotype, and EGFR -8227 G/A genotype are high risk factors for lung cancer in non-smoking female.

A 48-year-old man presented with a 4-day history of fever and 10-year history of papulovesicles on the face, neck, trunk and limbs which had been aggravated 10 days prior to the presentation.Skin biopsy showed a dermal infiltration of numerous small- to medium-sized atypical lymphocytes, which was mainly located around blood vessels or appendages, with the involvement of subcutaneous fat tissue and destruction of blood vessels. The infiltrating atypical cells stained positive for CD45RO, CD8, CD56, T-cell intracellular antigen-1, granzyme B, Epstein-Barr virus-encoded small nuclear RNAs (EBER), but negative for CD20, CD79a, CD3, CD4 or CD30. Cytoplasmic CD3ε was also observed in these cells. Laboratory examinations on admission revealed a progressive decrease in peripheral erythrocytes, white cells and platelets, persistent increase in serum aminotransferase and bilirubin, and decline in serum fibrinogen and hypertriglyceridemia. The B-mode ultrasound of the abdomen showed hepatosplenomegaly. Based on the above findings,the diagnosis was made as extranodal nasal type NK/T-cell lymphoma of skin complicated by hemophagocytic syndrome.

Objective To study the expression and the mechanism of miR-155in the villi of patients with unexplained recurrent spontaneous abortion (URSA).Methods The expression of miR-155 in the villi of 36 cases with URSA (URSA group) and 25 women with normal early pregnancy (control group) were detected by stem-loop real-time reverse transcription (RT) qPCR.Expression of hypoxia inducible factor-1 (HIF-1α),vascular endothelial cell growth factor(VEGF) and micro lymphatic vessel density (MVD) in the villi of were measured by immnohistochemical staining among two groups.Results (1) miR-155 expression:the mean miR-155 expression were 1.456 (0.489,2.459) in URSA group and 2.833 (1.740,3.794) in control group,which reached statistical difference (P ＜0.05).The mean expression of miR-155 of 1.683 (0.902,2.459) in URSA group with abortion times (≤ 3) was significantly higher than 1.229 (0.489,1.719) in URSA group with more than 4 times abortion (P ＜ 0.05).(2) Indexes:the expression of HIF-1α,VEGF and MVD value were 121 ± 12,134 ± 12,36 ± 6 in URSA group and 99 ± 10,109 ± 10,28 ±4 in control group,which reached statistical difference(P ＜ 0.01).The expression of HIF-1α,VEGF and MVD value of 119 ± 12,134 ± 12,35 ± 5 in URSA group with less than 3 times abortion was significantly lower than 128 ± 12,138 ± 12,43 ± 6 in URSA group with more than 4 times abortion (P ＜ 0.01).Conclusions The expression of miR-155 and HIF-1α is topically stimulated by oxygen signal.HIF-1α adjusts the transcription and translation of VEGF,which together involved in placental trophoblast invasion and placental angiogenesis.The low expression of miR-155 could interfere with expression of HIF-1α and VEGF,which might be involved in villous vascular dysplasia in URSA.

Objective To detect the prevalence of human cytomegalovirus (HCMV) in peripheral blood of patients with systemic lupus erthematosus (SLE) and explore its role in the pathogenesis of SLE.Methods HCMV DNA was isolated from the peripheral blood leucocytes (PBLs) of 60 patients with SLE and 111 healthy controls.Nested polymerase chain reaction (nPCR) technology was used to investigate the gene of HCMV glycoprotein gB (UL55) in these specimens.HCMV infections in the PBLs of SLE patients were confirmed by HCMV-UL55 detection.Two-sample t test,nonparametric test,Chi-square test and Fisher probabilities were used to analyze.Results Agarose gel electrophoresis and sequencing analysis showed that established nPCR could specifically detect HCMV-UL55 gene,the HCMV infection rate was significantly higher in patients with SLE than in the healthy controls (P＜0.01).Positive rates of HCMV infection in SLE group and controls were 41.7％ (25/60) and 1.8％ (2/111),respectively.Compared to the SLE patients with HCMV-negative PBLs,the positive rate of Rib-P [26％(9/35) vs 56％(14/25),x2=5.659,P=0.017],the positive rate of direct Coomb's test [37％(13/35) vs 72％(18/25),x2=7.096,P=0.008] and the level of antiβ2GP1 [21.3 (9.9,51.8) U/ml vs 13.6 (5.9,23.1) U/ml,U=2.017,P=0.044] were significantly higher than those in the SLE patients with HCMV-positive PBLs.Compared to the SLE patients with HCMV-negative PBLs,the number of red blood cells [(3.65±0.10)×1012/L vs (3.17±0.17)×1012/L,t=2.574,P=0.013] and lymphocytes [(1.37±0.14)×1012/L vs (0.90±0.13)×1012/L,t=2.456,P=0.017] in peripheral blood and the hemoglobin levels [(110±19) g/L vs (98±5)g/L,t=2.034,P=0.048] of the SLE patients with HCMV-positive decreased significantly.At the same time,the positive rate of hematuria [40％(14/35) vs 72％(18/25),x2=6.000,P=0.014] and 24 h proteinuria [0.80 (0.53,2.37)g vs 0.48 (0.13,1.21)g,U=2.140,P=0.032],which indicated kidney damage were also significantly increased in SLE patients with HCMV-positive PBLs.Conclusion The infection of HCMV in peripheral blood cells may take part in the development of SLE.

Objective To study the relationship between cardiovascular diseases (CVD)and 24-h peritoneal protein losses (PPL) in continuous ambulatory peritoneal dialysis (CAPD)patients. Methods One hundred and seventy-eight CAPD patients in our department were enrolled in this study. Their 24-h PPL was measured and other clinical data were recorded at the beginning. Meanwhile, Doppler ultrasound examination was performed. They were then followed-up prospectively for the development of CVD. Results The average of 24-h PPL was (5.0±1.8) g.Patients with diabetic status or preexisting CVD or carotid arteries arteriosclerosis had higher 24-h PPL than those without (t=2.082, P=0.039; t=2.601, P=0.010; t=2.217, P=0.029). 24-h PPL was positively correlated with left ventricular end-diastolic diameter (LVDd), interventricular septal thickness (IVSTd), posterior wall diameter of left ventricle at end-diastolic (LVPWd) and left ventricular mass index (LVMI) (r=0.222, P=0.040; r=0.217, P=0.043; r=0.339, P=0.002; r=0.305, P=0.007). It was negatively correlated with ejection fraction of left ventricle (r=0.221, P=0.040). One hundred and fourteen CAPD patients were prospectively followed-up for at least twelve months. Patients developing CVD were 40.4% and 19.3% for high and low PPL groups respectively (x2=6.035, P=0.014). In the multivariable logistic regression analysis, the 24-h PPL was one of the independent factors for developing CVD. Conclusions There is a significant and independent relationship between 24-h PPL and new cardiovascular events. 24-h PPL may be an important predictor of cardiovascular disease.

Objective To research the homology and cross reaction characteristics of human papillomavirus(HPV)16 type L2 N-terminal(1-200)protein in clinical common HPV infection types.Methods The amino acid sequences of the common HPV infection types(6,11,16,18 ,etc.)were blasted and it was found that 1-200 N-terminal sequence of L2 protein was highly homologous.The gene of HPV16 L2(1-200)was amplificated from tissue sample of cervical cancer patient and inserted into the prokaryotic expression vector PGEX-4T-1 to construct the recombinant plasmid PGEX-4T-1-HPV16 L2(1-200).After sequencing identification,the recombinant plasmid was tranformed into E.coli BL21(DE3).Induced by IPTG,the fusion protein containing HPV16 L2(1-200)was expressed and analyzed by both SDS-PAGE and Western blot.Furthermore,the specific binding capacity of the fusion protein to the HPV 6,11,16 and 18 DNA positive patient serums were analyzed by Western blot.The fusion protein was purified with Ni-NTA Agarose Kit and coated with ELISA reaction plates.The specific serum IgG of 98 condyloma acuminatum patients,135 cervix cancer patients and 96 healthy control subjects were detected respectively by indirect ELISA.Results After comparing the amino acid sequences of the common HPV infection types(HPV6,11,16,18,etc.),We found that the homology of HPV L2(1-200)reached 52.7%-74.3%.The recombinant plasmid PGEX-4T-1-HPV 16 L2(1-200)was constructed successfully.Highly expressed HPV16 L2(1-200)fusion protein was obtained and the expression level was account for up to about 22.6% of total bacterial protein.The relative molecular mass(Mr)of the fusion protein is about 49×103,which matches up to the expected Mr Meanwhile,the serums of HPV 6,11,16,18 DNA positive patients were used as the first antibody and the specific band was detected respectively at about 49 × 103 by Western blot.Indirect ELISA showed that the A490 values of the specific IgG of condyloma acuminatum group,cervical cancer group and healthy control subjects were 0.753 ± 0.262,0.756 ± 0.274 and 0.178 ± 0.157 with the positive rate were 89.8%,88.9% and 9.4% respectively.There was no significance of the specific IgG between condyloma acuminatum group and cervical cancer group(P＞0.05),but it was significant among the three groups(P＜0.001).Conclusion The N-terminal 1-200 amino acids of HPV L2 has high homology and there exits cross reaction among the most common HPV infection types.

Objective To investigate the expression of interferon-induced protein 44 (IFI44) gene in the leukocytes of the peripheral blood samples from patients with systemic lupus erythematosus (SLE), and to evaluate the relationship between the expression level and disease activity. Methods Mononuclear cells in the peripheral blood samples from 100 SLE patients were compared with those of 40 disease controls and 40 healthy donors (HD) and the expression of the IFI44 was evaluated by quantitative real-time PCR.Comparisons between groups were performed with ANOVA, and the correlation analysis between the level of expression was higher in SLE patients than disease controls and healthy donors (26.8±5.3, 7.4±2.7, 5.2±2.0,respectively) (P=0.0012, P=0.005), but no difference was found between disease controls and healthy donors. Mild disease activity and the SLE patients with stable disease (63.1±22.4, 28.0±7.2, 9.2±1.8, respectively)and 24 hours urine protein level (r=0.42, P=0.000). Conclusion IFI44 is demonstrated to be highly expressed in SLE patients. The level of IFI44 may be a promising candidate biomarker for identifying SLE activity.

Objective To evaluate the role of miR-143, miR-145 in the development of gastric gastrointestinal stromal tumor. Methods The expression levels of miR-143 and miR-145 in 21 cases of gastric gastrointestinal stromal tumor and the matched non-tumor adjacent tissue specimens were examined by stem-loop real-time RT-PCR, and its correlation with clinicopathologic features of gastric gastrointestinal stromal tumor were analyzed. Results Expression level of miR-145 were significantly higher in tumor than adjacent normal tissues (P＜0.01 ) and that with mitotic count ≥ 5/50HPF cases was significantly lower than that with mitotic count ＜5/50HPF cases (P=0.02). miR-145 expression in huge tumor (＞10 cm)was significantly lower than that in the large tumor (5～10 cm) and small tumor (2～5 cm) (P=0.048).By Fletcher risk stratification system, miR-145 expression in high-risk cases was significantly lower than that in the intermediate-risk and low-risk cases (P=0.048). While the expression of miR-145 in low-risk group was significantly different compared to that in intermediate-risk group and high-risk group (P=0.01).There was no difference between the expressions of miR-143 in tumor and that in normal tissue(P=0.06).Conclusion In gastric gastrointestinal stromal tumor, MiR-145 expression is significantly higher in tumor than adjacent normal tissues. miR-145 is closely associated with tumor size. mitotic counts and Fletcher risk stratification system.

PPARγ2 gene C1431T polymorphism was assayed by PCR-RFLP in 200 polycystic ovary syndrome( PCOS)patients and 150 normal subjects. Serum adiponectin and leptin levels were determined by ELISA method. Polymorphism of the site might be associated with serum leptin and adiponectin concentrations and thiazolidinedione treatment in women with PCOS ( P<0.05 or P<0.01).