Objective To investigate the association of PPAR gamma Pro12Ala polymorphisms with conroy heart disease(CHD). Methods One hundred eighty impatients were recruited in the study. Venous blood samples were collected after 12 h of fasting from all patients. DNA was extracted and Pro2Ala polymorphism in the PPAR gamma 2 genes was genotyped through the PCR-RFLP. The frequency and distribution of Pro2Ala polymorphism in Chinese Han population with CHD were analyzed. Gensini Score based on coronarography were used to grade the coronary. Results There were a total of 135 CHD patients,of which 52 were affected with single coronary lesion,43 with multi-coronary lesion,and 45 were normal. The frequency of P12/P12, A12/P12 in the coronary lesion group were 92.6% (125/135) and 7.4% (10/135) which were similar to that of 93.3% (42/45) and 6.7% (3/45)(P =0. 548). In the single coronary lesion group, the frequencies of P12/P12 and A12/P12 were 94.0% (42/52)and 9.6% (5/52) ,and 90.0% (36/40) and 10.0% (4/40) in the bi-coronary lesion group,97.7% (42/43) and 2.3% (1/43) in the multi-coronary lesion group,with no significant difference among the three groups( P ＞0.05).There were no significant difference on the frequency of P12/P12 and A12/P12 between various subgroups defined according to Gensini score (P = 0. 246). Compared to the frequency in non-obese, the frequency of A12/P12 were significantly higher in the obese ( P = 0.028 ). In PPAR gamma 2 genes B the Exton BstuI enzyme restriction site genes, BMI,waist-hip ratio and total cholesterol in the carriers of P12A genotype were significantly higher than those in the P12P genotype carriers ( P ＜ 0. 05 ). Conclusions PPAR gamma Pro12Ala polymorphisms might not be associated with CHD in Chinese Han population. The frequency of A12/P12 in obese is significantly higher than that in non-obese. PPAR gamma Pro12Ala polymorphisms might be associated with adiposity and lipid metabolism.

Objective To investigate the protective effect of Edaravone on dopamine transporter in rat models of Parkinson disease.Methods Rat models of Parkinson disease were induced by injection 6-OHDA into right medial forebrain bundle. Edaravone at different doses (3.0 mg/kg, 1.0 mg/kg or 0.3 mg/kg) was injected intraperitoneally twice daily for two weeks. The same dose of normal saline was injected in the control group. One week after the treatment, the ?-radiation of rat bilateral striatum, cerebral cortex and cerebella cortex of each group was measured by a ?-counter and the brain tissue ID value was calculated.Results There was a significant difference of the radiation count in right striatum between the large dose group (0.47?0.06) ,medium doss group(0.37?0.02)and the control group (0.25?0.01)( P

Objective To investigate the effects of the leflunomide active metabolite (A771726) on the expression of phorbol-12-myristate-13-acetate (PMA) -induced CD147, matrix metallo-proteinase (MMP)-2 and MMP-9 on THP-1 cells. Methods THP-1 cells were cultured in RPMI-1640 medium supplemented with 10％ fetal bovine serum. For all experiments, THP-1 cells were cultured at an initial density of 5×105/ml. Before A771726 treatment, cells were cultured with serum-free RPMI-1640 medium for 12 h, THP-1cells were co-cultured with PMA at three different concentrations of A771726 (5, 15 , 45 μg/ml) for 24 h.The mRNA expression of CD147, MMP-2 and MMP-9 was measured by real-time PCR. CD147 expression on the cells were evaluated by flow cytometric analysis. The activity of MMP-2 and MMP-9 were evaluated by gelatin zymography. Statistical differences among the groups were tested by one-way ANOVA or KruskalWallis test. Results The expression of CD147, MMP-2 and MMP-9 were upgraded by the PMA. The expression of CD147 on THP-1 cells was inhibited significantly by A771726 in a dose-dependent pattern (P＜0.01). The mean fluorescence intensity (MFI) of CD147 in positive control group was 109.5±3.8, the MFI in A771726 (5, 15, 45 μg/ml) group were 73.3±2.5, 64.5±2.3, 40.9±2.7, respectively. The expression of MMP-2, MMP-9 mRNA and the activity of MMP-2, MMP-9 in the supernatant was inhibited significantly by A771726 (P＜0.01). The expression of CD147 mRNA was not inhibited significantly by A771726 (P＞0.05).Conclusion Leflunomide active metabolite (A771726) can inhibit the expression of PMA-induced CD147,MMP-2 and MMP-9 on THP-1 Cells.

Objective To investigate the value of ultrasound in differential diagnosis of benign and malignant thyroid lesions with rimlike peripheral calcification.Methods Seventy-three patients of thyroid nodules with rimlike peripheral calcification were analyzed retrospectively.All cases were confirmed by surgery and pathology.The efficacy of sonographic features on diagnosis of thyroid nodules was analyzed.Results Among 73 patients,58 (58/73,79.45％) were benign and 15 (15/73,20.55％) were malignant.Among the sonographic features mentioned,the mean size,margin,internal echo and presence of halo showed no significant differences between malignant and benign nodules (all P＞0.05).Proportion of thyroid nodules coexisting with nodular goiter,irregular thickness and interruption of rimlike peripheral calcification had significant differences between malignant and benign nodules (all P＜0.05).The sensitivity,specificity,positive predictive value and negative predictive value of coexisting with nodular goiter for diagnosing benign nodules were 77.59％ (45/58),60.00％ (9/15),88.24 ％ (45/51),40.91％ (9/22),respectively.The sensitivity,specificity,positive predictive value and negative predictive value of irregular thickness for diagnosing malignant nodules were 53.33 ％ (8/15),87.93％ (51/58),53.33％ (8/15),87.93％ (51/58),respectively.The sensitivity,specificity,positive predictive value and negative predictive value of interruption of rimlike peripheral calcification for diagnosing malignant nodules were 73.33％ (11/15),68.97％ (40/58),37.93％ (11/29),90.91％ (40/44),respectively.Conclusion Ultrasonography is helpful to diagnosis of thyroid nodules with rimlike peripheral calcification.Irregular thickness and interruption of calcification are associated with malignancy.

BACKGROUND: Lentivirus-mediated RNA interference technology has been widely used in the study of gene function because of its specificity, high inhibition efficiency and persistent effects. Virus packaging, transfection, as well as small hairpin RNA (shRNA)-coding sequence could affect the inhibition efficiency. Therefore, impact factors of RNA interference for human Bax inhibitor 1 (BI-1) was studied in this experiment.OBJECTIVE: To find the valid small interference RNA (siRNA) targeting human BI-1 gene by Lentivirus.DESIGN, TIME AND SETTING: Single sample observation was completed in Department of Medical Genetics, Basic Medical College, Peking University Health Science Center from September 2007 to December 2008.MATERIALS: 293T cell and SH-SY5Y cells were preserved in this laboratory; 4 shRNAs were designed through a use of online design tool by Ambion (www.ambion.com). METHODS: Several recombinant plasmids which expressed shRNAs targeting different regions of the BI-1 gene and labeled enhanced green fluorescent protein as a fusion gene were constructed. Four types of shRNA-expressing virions were obtained by cotransfection of shRNA-expressing plasmid and package protein-expressing plasmids into 293T cells. Green fluorescent protein expression was determined using flow cytometry to search the optimal package conditions. Real-time PCR and beta actin mRNA was used as an internal control, four types of recombinant viral supernatant and control viral supernatant were added on the SH-SY5Y cells, knock-down efficacies of the endogenous BI-1 gene were determined, and the RNA interference effective sequences were optimized. MAIN OUTCOME MEASURES: Intracellular green fluorescent protein expression rate of the cells infected with different packaging virus. RESULTS: The optimal infective packaging virus could be obtained when the ratio of plasmids pLentiLox3.7, REV, VSVG and RRE was 2:1:1:1 and the supernatants were collected 48 hours after transfection. Replacement of culture media 24 hours after transfection could increase the infection efficacy of the virions. shRNA targeting-2-17nt (start coding region) of BI-1 gene was most valid interference efficiency and could decrease the gene expression to 40%. CONCLUSION: Influencing factors of Lentivirus-mediated RNA interference technology virus packages were investigated in this study, which could be a foundation for constructing stable BI-1 knocked down neuronal cell model and for studying the abnormal expression of BI-1 related to the neuron apoptosis disease.

Objective To investigate the pattern of fluctuations and clinical significance of fasting serum insulin(FINS) and C-peptide(CP) levels in patients with different subtypes of acute cerebral infarct(ACI) and its relationship with serum lipid. Methods FINS and CP were measured in 152 ACI patients by chemiluminescent immunoassay. All ACI patients were classified into 5 major ischemic stroke subtypes according to the trial of org10172 in acute stroke treatment(TOAST) criteria. And then the relationship between FINS and CP level and serum lipid in different TOAST subtypes were analysed. Results The percentage of each ischemic stroke TOAST subtype was as follow: stroke of undetermined etiology 40.13%, small-vessel occlusion 34.21%, cardioembolism 5.26%, large-artery atherosclerosis 15.79%, and stroke of other determined etiology 4.61%. Among 5 major stroke subtypes, large-artery atherosclerosis patients had the highest levels of FINS and CP. The levels of FINS and CP in small-vessel occlusion were (8.237?5.144) ?U/ml and (1.761?0.975)ng/ml,respectively. Stroke of other determined etiology subtypes were associated with the lowest levels of FINS and CP. Apparently, other factors, such as TC, TG, LDL, SBP, DBP, age and HDL, could also affect the levels of FINS and CP in serum. Conlusions Levels of FINS and CP varied in different subtypes of ACI. There was a significant correlation among insulin resistance, hyperinsulinemia(HIS) and lipid metabolic abnormality in ACI.

Objective: To investigate the second-hand smoke behavior at home among smokers, so as to provide the reference for developing home tobacco control strategies. Methods: Permanent residents who were smokers and at the ages of 15 years and above were sampled from 10 streets (townships) in Xuhui District, Shanghai Municipality using the multi-stage random sampling and population-size proportional sampling methods in 2022. Demographic information, smoking status, awareness of second-hand smoke hazards and second-hand smoke behavior at home were collected by questionnaire surveys. Factors affecting second-hand smoke behavior at home were identified using a multivariable logistic regression model. Results: A total of 1 024 smokers were surveyed, including 769 males (75.10%) and 255 females (24.90%). The awareness of hazards of second-hand smoke was 33.59%; the awareness rate of second-hand smoke causing lung cancer in adults was the highest at 76.76%, while the awareness rate of second-hand smoke leading to premature birth and low birth weight in newborns was the lowest at 39.45%. There were 459 smokers with second-hand smoke behavior at home, accounting for 44.82%. Multivariable logistic regression analysis showed that occupation (the retired, OR=2.320, 95%CI: 1.276-4.218), frequency of smoking (often, OR=5.722, 95%CI: 3.977-8.231), smoking duration (a year and above, OR=10.089, 95%CI: 5.508-18.480), electronic cigarette use (occasionally, OR=2.994, 95%CI: 1.283-6.986), living with pregnant women or infants (no, OR=2.171, 95%CI: 1.367-3.448), family indoor smoking restrictions (no restriction, OR=13.926, 95%CI: 7.538-25.727) and awareness of second-hand smoke hazards (unknown, OR=1.562, 95%CI: 1.067-2.287) were the influencing factors for second-hand smoke behavior at home. Conclusion There were 44.82% smokers in Xuhui District with second-hand smoke behavior at home, which was influenced by occupation, living situation, smoking status, family indoor smoking restriction and awareness of second-hand smoke hazards.

Objective To evaluate morphological structure of uterosacral ligament (USL) and cardinal ligament (CL) in patients with severe pelvic organ prolapse (POP) by MRI technology, and to analysis and discuss its clinical significance. Methods From November 2013 to February 2014 in Peking University People′s Hospital, 26 elderly patients withⅢ-Ⅳdegree of POP were selected as the POP group and 18 healthy elderly volunteers were selected as the control group during the same period. Pelvic MRI examination were performed in the two groups. The morphological characteristics of left and right side of the uterosacral-cardinal ligament on MRI and the attachment site of the starting and ending points between two group were described and compared. Results In POP group, 25 cases of left USL starting point were located in the sacrospinous ligament/coccygeal muscle complex [58% (15/26)] or coccygeal muscle [38%(10/26)], ending point were located in the cervix and vagina [58%(15/26)] or cervix [38%(10/26)];24 cases of right USL starting point were located in the sacrospinous ligament/coccygeal muscle complex [31%(8/26)]or coccygeal muscle [62%(16/26)], 26 cases of right USL ending point were located in the cervix and vagina [62% (16/26)] or cervix [38% (10/26)]; the left and right CL in the POP group and the control group were both from the sacroiliac joint at the top of the greater sciatic foramen from the ipsilateral pelvic side wall;1 case (4%, 1/26) of left CL in the POP group completely connected to the bladder, 10 cases (38%, 10/26) partly connected to the bladder;14 cases (54%, 14/26) of right CL partly connected to the bladder, the rest ending points of left and right CL were located in cervix and (or) vagina. In the control group, 17 cases of left USL starting point were located in the sacrospinous ligament/coccygeal muscle complex (10/18) or coccygeal muscle (7/18), ending point were located in the cervix and vagina (12/18) or cervix (6/18);18 cases of right USL starting point were located in the sacrospinous ligament/coccygeal muscle complex (10/18) or coccygeal muscle (8/18), ending point were located in the cervix and vagina (13/18) or cervix (5/18);8 cases (8/18) of left CL partly connected to the bladder;15 cases (15/18) of right CL partly connected to the bladder, the rest ending points of left and right CL were located in cervix and (or) vagina. There was no significant difference between the two groups on the starting and ending points (P>0.05). Conclusions The observation of MRI could be consistent with the clinical anatomy on the starting and ending points, direction of travel in the uterosacral-cardinal ligament. The starting and ending points of the left and right side USL and the ending points of the left and right side CL are not completely symmetrical, the variation degree is large, some CL could be completely or partly inserted to the bladder.

Objective To investigate the renoprotective effect of transforming growth factor beta activator kinase 1 (TAK1) inhibitor 5Z-7-oxozeaenol (OZ) in diabetic db/db mice and the mechanism.Methods Twenty-four male db/db mice were randomly divided into two groups:db/db mice (db/db,n=12) and db/db mice with 5Z-7-oxozeaenol treatment (db/db+OZ,n=12).Another group of wild type mice (n=12) was held as the control group.OZ 2 mg/kg was administrated by intraperitoneal injection every other day.At week 8 and 12 after 5Z-7-oxozeaenol treatment,blood glucose (BG),body weight (BW),kidney weight (KW) and urinary albumin excretion rate (UAER) were evaluated.Kidney pathological lesions were detected by light and electron microscopy.NF-κB p65,monocyte chemotactic protein-1 (MCP-1) and tumor necrosis factor-ot (TNF-o) were detected by immunohistochemistry.Western blotting was used to detect p-TAK1,TAB1,p-p38MAPK and IL-1β expression,while ICAM-1 and MCP-1 mRNA levels were evaluated by real-time PCR.Results Compared with control group,the levels of BG,BW,KW and UAER were higher (P ＜ 0.01) in db/db mice group,while BW,KW and UAER levels were significantly decreased in db/db + OZ group compared with that in db/db mice group (P ＜ 0.05).In week 8 and 12 db/db mice,glomerular volume and extracellular matrix were increased,while pathological lesions in kidney tissue were positively improved by TAK1 inhibitor.Immunohistochemistry showed that NF-κB p65,MCP-1 and TNF-α expression levels were apparently increased in db/db mice group compared with that in control group (P ＜ 0.05) and were significantly inhibited by TAK1 inhibitor (P ＜ 0.05).Western blotting showed that p-TAK1,TAB1,p-p38MAPK and IL-1β expression levels were higher in db/db mice group than that in control group (P ＜ 0.05) and lower in db/db+ OZ group than that in db/db mice group (P ＜ 0.05).Moreover,real-time PCR showed that the expressions of ICAM-1 and MCP-1 mRNA were higher in db/db mice group than that in control group and lower in db/db+OZ group than that in db/db mice group (P ＜0.05).Conclusions TAK1 Inhibitor can down-regulate MAPK and NF-κB pathway to restrain the reaction of inflammation and alleviate kidney injury in diabetic db/db mice.

Objective To investigate the role of transforming growth factor-β activated kinase-1 (TAK1) signaling pathway in the activation of bone marrow derived macrophages (BMDM) induced by high glucose.Methods Purity of mouse BMDM was detected by flow cytometry.The mice macrophages cultured in vitro were stimulated by high glucose and treated with TAK1 specific inhibitor 5Z-7-oxozeaenol.Cells were divided into normal control group (RPMI 1640),osmolality control group (25 mmol/L mannitol),high glucose group (33 mmol/L D-glucose) and inhibitor group (33 mmol/L D-glucose+300 nmol/L 5Z-7-oxozeaenol).Immunocytochemistry and flow cytometry were used to detect macrophage subtype.The expression of monocyte chemotactic protein-1 (MCP-1) and tumor necrosis Factor-α (TNF-α) mRNA were determined by real time PCR.Expressions of p-TAK1,TAK1 binding protein (TAB1),p-JNK,p-p38 MAPK and NF-κB p65 proteins were analyzed by Western blotting.Results The purity of BMDM was about 99.36％.Compared with normal control group,high glucose group had increased percentage of M1 macrophages,increased expression of MCP-1 and TNF-α mRNA (all P ＜ 0.05).Moreover,p-TAK1,TAB1,p-JNK,p-p38 MAPK and NF-κB p65 proteins expression also increased significantly in high glucose group (all P ＜ 0.05).After treatment with inhibitor 5Z-7-oxozeaenol,the effects induced by high glucose were inhibited (P ＜ 0.05).Conclusions High glucose can induce M1 macrophage activation and expression of inflammatory cytokine of BMDM,which can be inhibited 5Z-7-oxozeaenol through inhibiting TAK1/MAPK and TAK1/NF-κB pathway.