[Summary] A selection of 528 unrelated subjects were enrolled(198 males, 330 females) with the mean age of(52. 23 ± 13. 41) years old. According to body mass index, 253 persons belonged to the normal weight group and 275 persons overweight/ obesity group. A total of 10 SNPs in CIDEB and CIDEC genes were detected. SHEsis online were used to get the haplotypes of these two genes. The relationship between above SNPs and obesity were analyzed under additive inheritance pattern. The main effects and interaction on obesity induced by two genes’ haplotypes were analyzed by logistic regression. rs2144493 in CIDEB gene was associated with obesity, C was a protective alleles, OR (95% CI) equals 0. 722(0. 525-0. 992). CCTT haplotype of CIDEB gene carriers and GCG haplotype of CIDEC gene carriers were more prone to obesity or overweight, there was an interaction between the haplotypes of 2 genes. CIDEB, CIDEC haplotypes may play independent and interactive roles in causing obesity.

Objective To investigate and evaluate the diagnostic value of transcranial doppler ultrasound (TCD) on brain death in patients with severe craniocerebral injury.Methods Forty-two cases of severe craniocerebral injury from Affiliated Hospital of Jining Medical University were divided into brain death group and survival group,according to the clinical prognosis.TCD was conducted to examine brain blood flow numerical and changed characteristics of the spectrum.Average blood flow velocity (Vm),pulse index (PI),diastolic blood flow in reverse (reverberating flow) and small systolic peaks in early systole (wave of spikes)were documented.TCD parameters were compared between the two groups with t test.Results In the brain death group,frequency of reverberating flow was 86.67％ and frequency of wave of spikes was 13.33％.In survival group,frequency of reverberating flow was 14.81％ and no wave of spikes was recorded.The difference in frequencies of reverberating flow and wave of spikes between the two groups was significant(x2 =28.25,P ＜0.01).The average speed of bilateral middle cerebral artery blood flow in brain death group((20.02±13.96) cm/s) was significantly reduced compared with survival group((56.81± 16.84) cm/s,t=2.30,P＜0.05).Pulse index (PI) values in brain death group (4.02±3.49)were significantly increased compared with survival group (1.24±1.03) (t=2.10,P＜0.05).Conclusion Reverberating flow or wave of spikes is an important marker for brain death in patients with severe craniocerebral injury.TCD may be applied to clinically confirm the diagnosis of brain death.

objective To assess the effect of aldosterone on the production of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9)and collagen Ⅳ in culture supematants of podocytes and the possible molecular mechanisms involved in the influence of aldosterone on the synthesis and degradation of extracellular matrix produced by podocytes. Methods Podecytes were treated with aldosterone at the concentration of 10-11, 10-9, 10-7 mol/L respectively. Cultured podocytes were examined at 24, 48 and 72 hours respectively. Spironolactone, a receptor antagonist of aldosterone, was added to observe the blocking effect on aldosterone. An inhibitor of TGF-β1 receptor was used to determine whether the effect of aldosterone on podocytes were mediated through TGF-β1 system. The enzymatic activities of MMP-2 and MMP-9 were assayed by gehtin zymography. Collagen Ⅳ 0.5 chain and TGF-β1 proteins released into culture supematants were assessed by Western blot and ELISA analysis. The adhesion rate of podocytes was monitored by flow cytometry. Results Aldosterone increased the activities of MMP-2 and MMP-9 in a dose- and time-dependent manner (P<0.05). Aldosterone decreased the level of collagen Ⅳ or5 chain protein in culture supernatants (P<0.05). Meanwhile, the expression of TGF-β1 was also increased (P<0.05). Spironolactone completely abolished the above-mentioned changes(P< 0.05). Blockage of TGF-β1 signaling with SB431542 prevented the aldosterone-induced upregulation of MMP-2 and MMP-9 as well as the downregulation of the collagen Ⅳ α5 chain protein and the adhesion rate of podocytes (P<0.05). Conclusions Aldosterone increases the activities of MMP-2 and MMP-9 but decreases the expression of collagen Ⅳ α5 chain and the adhension rate of podocytes possibly via TGF-β1 signaling pathway. Such alterations may contribute to glomerular podocyte injury associated with the GBM abnormality caused by the imbalance between matrix synthesis and degradation.

To investigate the effects of albumin on the production of matrix metalloproteinases-2 (MMP-2) and matrix metalloproteinases-9 (MMP-9) in podocytes. Podocytes were treated with bovine serum albumin (BSA) at the concentration of 0.1, 0.5, 1, 2 g/L, respectively. Conditioned media were harvested 12, 24, 48 and 72 h after the treatment. The expression of MMP-2 and MMP-9 was assayed by gelatin zymography, RT-PCR and Western blotting analysis. Our results showed that in comparison with the control group, BSA increased the expression of MMP-2 and MMP-9 mRNA and protein in a dose- and time-dependent manner (P<0.05). Meanwhile, the enzymatic activities of MMP-2 and MMP-9 in the culture supernatants of podocytes were also increased (P<0.05). It is concluded that albumin up-regulated the expression of MMP-2 and MMP-9 at gene and protein levels in a time- and dose-dependent manner.

Eukaryotic expression vectors carrying the small hairpin RNA (shRNA) for TRPC6 mRNA were constructed, and the effects of knocking-down TRPC6 on puromycin aminonucleoside (PAN)-induced apoptosis of mouse podocytes were observed. Two eukaryotic expression vectors containing small hairpin structure targeting TRPC6 named pGCsi-TRPC6A and pGCsi-TRPC6B were designed and synthesized. The plasmids were transfected into conditionally immortalized murine podocyte cell line by liposome. The changes in the TRPC6 mRNA and protein expression were observed by RT-PCR and Western blot after 48 h. Cultured podocytes were divided into four groups: control group, PAN treatment group, PAN treatment+shRNA transfection group, and PAN treatment+ negative control group. The expression of Bax and Bcl-2 mRNA and proteins was detected by RT-PCR and Western-blot respectively. The apoptotic rate of podocytes was measured by flow cytometry. The results showed that the expression of TRPC6 mRNA and protein was decreased in the podocytes when transfected with pGCsi-TRPC6A, and pGCsi-TRPC6B. The expression of Bax was increased, and that of Bcl-2 was decreased at protein and mRNA levels in the podocytes after treated with PAN for 48 h. These changes was attenuated by knocking-down TRPC6. Knocking-down TRPC6 could effectively decrease the PAN-induced apoptosis of podocytes. It was concluded that TRPC6 may play an important role in the PAN-induced apoptosis of podocytes. Knocking-down TRPC6 gene could effectively prevent the podocytes from apoptosis induced by PAN.

To investigate the protective effects of eplerenone on adriamycin-induced renal injury and the possible mechanisms involved, 36 male Sprague-Dawley rats were randomly divided into control group, adriamycin nephropathy (AN) group and eplerenone-treated group (100 mg·kg(-1)·d(-1) eplerenone). Blood pressure, 24-h urinary protein, serum potassium, sodium and creatinine were measured 28 days after adriamycin injection (a single tail intravenous injection of 6.5 mg/kg adriamycin). The morphological changes of renal tissues were observed by light and electron microscopy. Immunohistochemistry and Western blotting were performed to examine the expression of TGF-β(1) and desmin in renal cortex. The results showed that 28 days after adriamycin injection, there were no significant changes in the level of serum potassium, sodium, creatinine concentrations and blood pressure values in the rats of the three groups. Meanwhile, the 24-h proteinuria excretion in the AN group was significantly higher than that in the control group (P<0.01), but that in the eplerenone-treated group was substantially reduced when compared with that in the AN group (P<0.05). Mild mesangial cell proliferation and matrix expansion, diffuse deformation and confluence of foot processes in podocytes were found in the AN group. By contrast, rats in the eplerenone-treated group exhibited obvious attenuation of these morphological lesions. The protein expression of TGF-β(1) and desmin in the AN group was markedly up-regulated in contrast to that in the control group (P<0.01), whereas that in the eplerenone-treated group was much lower than in the AN group (P<0.05). It was concluded that eplerenone may ameliorate the proteinuria and the development of pathological alteration in adriamycin-induced nephropathy presumably via the inhibition of cytokine release, and restore the morphology of podocytes independent of its blood pressure-lowing effects.

Objective To investigate the relationship between obesity and esophageal high resolutionmanometry ,24‐hour pH monitoring and gastroscopic results of patients with gastroesophageal reflux disease (GERD) .Methods A total of 196 patients with GERD(DeMeester score>14 .72) were selected and divided into normal weight group (18 .5 kg/m2 < BMI < 24 kg/m2 ) , overweight group (24 kg/m2 ≤BMI<28 kg/m2 ) and obese group (BMI≥28 kg/m2 ) according to body mass index (BMI) . Esophageal high resolution manometry ,gastroscopy and 24‐hour pH monitoring were performed with DeMeester score calculated . The classification of esophagitis was according to Los Angeles standard . Normal distributed measurement data were compared by analysis of variance .Non normal distributed measurement data were repesent as M(P25 ,P75) ,and were compared by rank sum test .Chi square test was for count data comparison .Results Compared with normal weight group and overweight group , abdominal length of low esophageal sphincter (LES) of obese group was shorter (1 .90 cm ,0 .85 cm to 2 .45 cm ;2 .85 cm ,2 .23 cm to 3 .20 cm ;2 .50 cm ,1 .98 cm to 3 .0 cm ) , and the differences w ere statistically significant (Z=19 .913 ,P<0 .01) .But there was no significant difference in pressure ,total length of LES and distal esophagus amplitude (all P>0 .05) .The percent total time pH≤4 of obesity group was 15 .42% (10 .31% to 21 .49% ) ,percent supine time pH≤4 was 14 .21% (5 .75% to 34 .98% ) and percent upright time pH≤4 was 14 .25% (8 .19% to 18 .13% ) .The reflux episodes (106 .50 ,67 .00 to 145 .75) and the longest duration of reflux episodes (28 .10 min ,10 .90 min to 47 .93 min) were more than those of normal group (9 .74% ,5 .35% to 15 .96% ;7 .31% ,3 .25% to 11 .80% ;8 .45% ,5 .43% to 17 .48% ;72 .50 ,53 .00 to 100 .50;15 .80 min ,9 .90 min to 21 .28 min) and overweight group (11 .36% , 6 .74% to 15 .87% ;7 .74% ,2 .36% to 15 .05% ;11 .27% ,3 .37% to 14 .73% ;85 .50 ,58 .75 to 117 .75;21 .40 min ,11 .50 min to 39 .90 min) ,and the differences were statistically significant (Z=7 .054 ,11 .181 , 6 .429 ,6 .452 ,8 .246 ,all P<0 .05) .The incidences of hiatus hernia and reflux esophagitis of the obese group (both 56 .67% (17/30)) were both higher than those of normal weight group (36 .46% (35/96) and 30 .21% (29/96)) and overweight group (30 .00% (21/70) and 27 .14% (19/70)) ,and the differences were statistically significant (χ2=6 .439 and 9 .000 ,both P<0 .05) .However ,there was no statistically significant difference among the three groups in the incidence and severe degree of asthma as an extra esophageal appearance (all P>0 .05) .There was no statistically significant difference in the incidence of Barrett′s esophagus among three groups (all P>0 .05) .Conclusions Compared with that of normal weight group and overweight group of patiento with GERD ,abdominal length of LES of obesity group was shorter .With an increase in BMI , acid exposure and the incidences of reflux esophagitis and hiatal hernia also increased .

Objective To access the effects of aldosterone (ALD) and its receptor antagonist- spironolactone (SPI) on the production of reactive oxygen species (ROS) and apoptosis in podocytes and to explore the possible mechanism involved. Methods Conditionally immortalized mouse podocytes were divided into control group, ALD group, SPI group and SPI +ALD group. The level of ROS production and the expression of nephrin protein were assayed by fluorescent spectrophotometry and indirect immunofluorescence technology. The apoptosis rate of podocytes was monitored by flow cytometry. The expression of bax and bcl-2 mRNA and protein was detected by RT-PCR and Western blotting methods. The anti-oxidant N-acetylcysteine (NAC)was applied to determine whether the effects of ALD on podocytes were mediated by ROS pathway.Results Compared with the control group, ALD significantly increased ROS production in podocytes (P＜0.05). SPI completely abolished the above-mentioned effect of ALD (P＜0.05). ALD induced the down-regulation of the expression of nephrin and the up-regulation of podocytes apoptosis (P＜0.05), which was accompanied with the elevated expression of bax mRNA and protein and the reduced expression of bcl-2 mRNA and protein (P＜0.05). SPI or NAC prevented the above-mentioned changes induced by ALD (P＜0.05). Conclusion ALD increases theexpression of pro-apoptotic factor (bax) but decreases the expression of anti-apoptotic factor (bcl-2)to induce podocytes apoptosis through the mineralocorticoid receptor (MR) possibly via the mechanisms involving ROS pathway.

Objective To explore the association between the CpG methylation level of positive regulatory domain containing 16(PRDM16)gene promoter and obesity or body mass index(BMI). Methods A total of 116 patients(91 female adults and 25 male adults) with abdominal operation in a municipal hospital of Henan province were enrolled in this study and they were divided into two groups:normal weight group(n=50), overweight or obesity group ( n=66 ) . Fasting plasma glucose, total cholesterol, triglyceride, high density lipoprotein and low density lipoprotein were measured in peripheral blood. DNA was extracted from white blood cells in peripheral blood and modified by bisulphite. Then the CpG methylation level of PRDM16 gene promoter was detected by mass spectrometry. Finally, all data were analyzed by IBM SPSS Statistics 21. 0 at the 5% level. The essential features and biochemical indexes of research objects between two groups were compared by two independent sample t-test, except chi-square test for gender. The correlation between CpG methylation level of PRDM16 gene and BMI was analyzed by multiple linear regression. Results There were no significant differences ( P>0. 05 ) in the methylation levels of PRDM16 gene's effective CpG sites(including CpG5. 6, CpG8, CpG9, CpG12, CpG13. 14. 15, CpG26. 27, CpG28 and CpG29) between two groups. The methylation level of CpG26. 27 had positive linear relation with BMI in overweight or obesity group with the standardized coefficients of 46. 928(P=0. 015), which means the higher the methylation level is, the higher the BMI would be. Conclusion The CpG26. 27 methylation level of PRDM16 gene promoter region may have relationship with the risk of obesity.

poptosis induced by PAN.