Objective This study was designed to explore the effect of hyperglycemia on the expression of Toll-like receptor 4 (TLR4 ) in renal tubular epithelial cells and its significance in diabetic nephropathy. Methods In vitro cultured renal tubular epithelial cells ( NRK-52E) were divided into LG group (cultured in 5mmol / L glucose DMEM) and HC group (cultured in 25mmol / L glucose DMEM). Cells were harvested at different time points. Immunohistochemistry, Rt-PCR, Western Blot were used to detect TLR4 protein and mRNA expression, and the levels of IL-6 and TNF-α from the cell culture supernatant were determined by EL1SA assay. Results After 6 hours, there was increased expression TLR4 mRNA in HC group, which appeared to be maintained for 24 hours and began to decrease after 48 hours ( P < 0.05). TLR4 protein expression increased in HC group after 24 hours, and increased even further after 48 hours. Compared with LG groups, the difference had statistical significance ( P <0.05). In HG group, IL-6 and TNF-α expression in the supernatant from the NRK-52E culture were significantly increased ( P < 0.05) , and the expression of IL-6 and TNF-α was positive correlated with the expression of TLR4 protein ( r =0.799,0.820). Conclusion High glucose triggers an increase in expression of TLR4 in NRK-52E cells, itself leading to an increase in expression of inflammatory factors such as TNF-α and IL-6. In this way, TLR4 participates in the progress of diabetic nephropathy.

Objective To investigate the saponins from the Chinese materia medica Huluba (the seeds of Trigonella foenum-graecum L.). Methods To isolate and purify the single saponin by the column chromatography and dry column chromatography of silica gel H. The structural elucidation was carried on by spectroscopic evidence of (()~(13)C-NMR), FAB-MS, DEPT and the results of the fraction-hydrolysis of acquiring the secondary glucosides. Results The newly isolated saponin D was consisted by six molecules of sugar with diosgenin. The chemical structure of saponin D is: diosgenin-3-O-?-L-rhamnopyranosyl (1→3)-?-D-glucopyranosyl (1→4)-?-L-rhamnopyranosyl [(1→3)-?-L-rhamnopyranosyl] (1→4)-?-D-glucopyranosyl (1→4)-?-D-glucopyranoside. Conclusion Saponin D is a new one consisted of six molecules of su-(gar) with diosgenin.

Object To investigate the saponin from the seeds of Trigonella foenum graecum Linn (STFG) Methods The total saponin from STFG was extracted and purified by using the absorptive resin, the single saponin was isolated by using the column chromatography as well as dry column chromatography of silica gel H The chemical structure was elucidated by 13 CNMR , FAB MS, DEPT spectroscopic evidence and the results of fraction hydrolysis of acquiring their secondary glucosides Results A new saponin A from the total saponin has been obtained, the fraction hydrolysis carried out and the secondary glucoside Ⅰ and Ⅱ identified by determining the structure of saponin A The chemical structure of saponin A is: diosgenin 3 O ? L rhamnopyranosyl(1→4) ? D glucopyranosyl(1→4) ? D glucopyranoside The secondary glucoside Ⅰ is: diosgenin 3 O ? D glucopyranoside; Ⅱ is: diosgenin 3 O ? D glucopyranosyl(1→4) ? D glucopyranoside Conclusion Glucoside A is a new saponin with three molecules of sugar

Object To do detail investigation of the saponins from the Chinese materia medica Huluba (the seeds of Trigonella foenum graecum L ) Methods The pure saponins from the total saponins were isolated by employing the column chromatography and dry column chromatography of silica gel H Their chemical structures were elucidated by 13 C NMR , FAB MS, DEPT spectroscopic evidence and the results of the fraction hydrolysis of acquiring their secondary glucosides were obtained Results Two new saponins B and C were isolated and both were the glucosides consisted by four molecules of sugar with diosgenin The chemical structure of B is: diosgenin 3 O ? L rhamnopyranosyl (1→3) ? L rhamnopyranosyl (1→4) ? D glucopyranosyl (1→4) ? D glucopyranoside And saponin C is: diosgenin 3 O ? D glucopyranosyl (1→4) ? L rhamnopyranosyl (1→4) ? D glucopyranosyl (1→4) ? D glucopyranoside Conclusion Saponins B and C are two new ones with four molecules of sugar respectively

Objective:To analyze the chemical constituents of the ether extracts from the fruits of 3 species of Chaenomeles . Methods: The chemical constituents were analyzed by GC MS. Results: Sixty one constituents were firstly isolated and identified from the fruits of 3 species of Chaenomeles . Forty compounds in Chaenomeles lagenaria , thirty four compounds in Xuan mugua ( C.lagenaria from Xuanzhou) and 23 compounds in C.sinensis were isolated and identified respectively. There were 10 compounds in all 3 species of Chaenomeles , but their contents were very much different. They were acetic acid, benzaldehyde, n caproic acid, 14 methyl pentadecanoic methyl ester, glycerin, benzoic acid, 6,9 octadecadienoic methyl ester, squalane, stearic acid and oleic eicosyl ester. Conclusion: The composition and content vary with different species and district of Chaenomeles . The result may be useful for the evaluation of Chaenomeles fruits quality. [

Background Package and tissue patch implantation are common methods for repair of filtering bleb leaking after anti-glaucoma surgery.But the scarring or re-leakage of filtering bleb probably occur again.Objective This study was to investigate the repair effect of acellular dermal matrix (ADM) on filtering bleb leaking in rabbit model and compare the effectiveness among ADM,amniotic membrane and conjunctival overlap.Methods Trabeculectomy was performed on 48 eyes of 24 New Zealand rabbits,and models of filtering bleb leaking were established.The models were randomized into ADM group,amniotic membrane group and conjunctival covering group based on randomized number table.ADM patches with 4 mm×4 mm were implanted across lamellar cornea and sclera at a bridge in the ADM group,and the same size of amniotic membranes were used in the amniotic membrane group,and conjunctiva was sutured to limbus in the conjunctiva overlap group.The intraocular pressure (IOP) was measured before surgery and 1 day,1 week,1 month,3 months and 6 months after surgery.The biocompatibility of materials above was assessed under the slit lamp microscope,and the status of filtering bleb was evaluated and compared with anterior segment optical coherent tomography (AS-OCT) 1 day,1 week,1 month,3 months and 6 months after surgery.Results Before surgery and 1 day,1 week,1 month,3 months and 6 months after surgery,the IOP was (26.9±4.3),(16.6±5.1),(22.1 ±6.2),(18.3±6.5),(22.7±2.5),(23.4±1.4) mmHg in the AMD group,(29.9±5.4),(14.9 ± 6.4),(21.6 ± 7.8),(26.3 ± 4.1),(26.0 ± 4.2) and (23.0 ± 5.3) mmHg in the amniotic membrane group,and (28.7 ±4.3),(15.7 ±7.0),(22.0±6.3),(28.2±4.1),(24.7 ±4.1),(23.0±2.7) mmHg in the conjunctival overlap group,showing significant differences among different groups and various time points (Fgroup =8.419,P=0.011 ;Ftme =15.543,P=0.000).The IOPs were significantly lower from 1 day through 3 months after operation than those before operation in the AMD group (P =0.000,0.000,0.006,0.045) ; while the IOPs were reduced only from 1 day through 1 week after operation in comparison with before operation in the amniotic membrane group and the conjunctival overlap group (P =0.000,0.001).One month after surgery,the IOPs were significantly declined in the ADM group compared with the amniotic membrane group and the conjunctival overlap group (P =0.001,0.000).The grafts were clear under the slit lamp microscope and exhibited the valid filtering bleb until 3 months after operation under the AS-OCT in the ADM group.However,the valid filtering bleb remained only 1 month after surgery in the amniotic membrane group and the conjunctival overlap group.Neovascularization on the filtering bleb was found 3 months in the AMD group but 1 month in the amniotic membrane group and the conjunctival overlap group.Conclusions Compared with amniotic membrane and conjunctival tissue,ADM patch for the repair of filtering bleb leakage can increase the survival duration of filtering bleb and remain lower IOP.

Objective To determine the effect of aldosterone and its antagonist, spironolactone on epithelial-mesenchymal transition (EMT) of normal rat kidney epithelial cells (NRK-52E) in a high glucose milieu,and to explore the mechanism of renoprotection in diabetic nephropathy (DN ) in rats involving aldosterone and spironolacton. Methods NRK-52E cells were simultaneously cultured in the serum-free Dulbecco's modification of Eagle's medium Dulbecco (DMEM) for 12 hours. Then the low glucose (LG) group was cultured in LG (1000 mg/L) DMEM:The high glucose (HG) group was cultured in high glucose (4 500 mg/L) DMEM. The aldosterone (Aldo) groups were cultured in high glucose DMEM with the addition of 10,50 and 100 nmol/L aldosterone respectively. The SP group was cultured in high glucose (4 500 mg/L) DMEM plus 10~(-7)mol/L spironolactone. Immunohistochemistry, RT-PCR and Western blot were used to detect E-cadherin and α smooth muscle actin(α-SMA) mRNA expression. Results RT-PCR showed that compared with the LG Group, E-cadherin mRNA expression in the HG group was significantly lower, and α-SMA mRNA expression was significantly increased(P＜0.05). E-cadherin mRNA expression in the 50 nmol/L Aldo group and 100 nmol/L Aldo group was significantly lower than that in the HG group, while the expression of α-SMA mRNA was significantly increased in the HG group(P＜0.05), with a dose-dependent relationship with aldosterone(r=-0.70,P＜0.05;r=0.67, P＜0.05). E-cadherin mRNA in the SP group was significantly higher,while α-SMA mRNA expression was lower than that in the HG group(P＜0.01). Immunohistochemistry and Western blot showed that compared with the LG group, E-cadherin protein expression was significantly reduced, and α-SMA expression was significantly increased in the HG group(P＜0.01). In the 10 nmol/L Aldo, 50 nmol/L Aldo, and the 100 nmol/L Aldo groups, E-cadherin protein expression was significantly lower, and α-SMA protein expression was significantly higher than that in the HG group(P＜0.05), with a dose-dependent relationship with aldosterone(r=-0.83,P＜0.05;r=0.81, P＜0.05). In the SP group, E-cadherin protein expression was higher, and α-SMA protein level was lower than that in the HG group(P＜0.05). Conclusion Aldosterone can promote EMT of tubular epithelial cells in a high sugar milieu, leading to renal interstitial fibrosis in Diabetic nephropathy. Spironolactone can inhibit high glucose-induced renal tubular epithelial cells EMT, which may be an important mechanism for the inhibition of renal interstitial fibrosis.

Objectives To investigate the changes of cystatin C in the serum of patients with acute kidney injury ( AKI) or end-stage renal disease ( ESRD), and study its significance in the early diagnosis of AKI and its correlation with the degree of renal function injury. Method The cases in Xiangya hospital were enrolled in this study according to the RIFLE criteria, including 20 cases of slight acute kidney injury, 30 cases of medium-severe acute kidney injury, 48 cases of victims of the 5. 12 wenchuan earthquake, 32 cases of end-stage renal disease and 20 healthy patients. The microparticle-enhanced immunoturbidimetry method was used to detect serum cystatin C, and the colorimetric method was used to detect urine N-acetyl-beta-D-glucosaminidase (NAGase). The enzymic method was used to detect serum creatinine. The correlation between serum cystatin C and serum creatinine was analyzed, and the sensitivity and specificity of serum cystatin C were evaluated with the receiver operator characteristic curve (ROC curve). Results Compared with healthy control group, the serum cystatin C increased obviously in acute kidney injury group and ESRD group( P <0. 05 and P <0. 01). The serum cystatin C was positively correlated with serum creati-nine( P <0.01). The serum cystatin C in the 5. 12 wenchuan earthquake injured group was also higher than that in healthy control group ( P <0.05), an the serum cystatin C had an AUC - ROC of 0.931 ( P <0. 01). Conclusion Compared to the conventional biomarkers, the earlier emergence of serum cystatin C can contribute better to early clinical diagnosis of AKI. The serum cystatin C is positively correlated with renal function, and reflect changes in renal function accurately.

ET-CT demonstrated that VX2 tumor tissues could uptake 18F-FDG more than normal tissue, which made the basis for further study of VX2 tumor model.

To investigate the role of H-fas oncogene in the early stages of human breast carcinogenesis. Methods Thirty cases of human breast cancer, 36 epithelial hyperplasia of usual type and 31 atypical hyperplasia were employed to detect H-ras gene codon 12 mutations by PCR-RFLP and PCR-SSCP assays, and to detect the expression of H-ras protein by immunohistochemistry method. ResultsExpression of H-ras protein were found in 73.3 % (22/30) of breast cancer and 48.4 % (15/31 ) of atypical hyperplasia. No H-ras protein expression was observed in hyperplasia of usual type. All tested sarnples of breast cancer and hyperplsia showed no mutations of H-ras gene codon 12. ConclusionOverexpression of H-ras protein is involved in early stages of breast carcinogenesis, but mutations of H-ras gene codon 12 is rarely present in the stage.