AIM To develop an HPLC method for the simultaneous content determination of ginsenosides Rb1,Rd,F4,Rg1,20 (R)-Rg3,20 (S)-Rg3,Rgs,20 (R)-Rh1,20 (S)-Rh1,Rh4,Rk1,Rk3 and notoginsenoside R1 in Shusanqi Powder (processed Notoginseng Radix et Rhizoma).METHODS The analysis of 70％ methanol extract of this drug was performed on a 35 ℃ thermostatic Agilent Zorabax-C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-water flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 203 nm.RESULTS Thirteen saponins showed good linear relationships within their own ranges (r =0.999 5),whose average recoveries were 90.01％-108.32％ with the RSDs of 0.62％-3.54％.CONCLUSION This sensitive and accurate method can be used for the quality control of Shusanqi Powder.

Objective To investigate the protective effects on the renal allografts from brain dead (BD) donor rats pretreated with bone marrow mesenchymal stem cells (MSCs).Method Three groups [normal transplant group (G1).BD transplant group (G2),and MSCs pretreated + BD transplant group (G3)] were set up.Male F344 rats served as donors and male Lewis rats as recipients.In G1,kidneys from F344 donor rats were implanted into Lewis recipients.In G2,kidneys from F344 BD donor rats were engrafted into Lewis recipients.In G3,after BD was established in F344 rats,MSCs were given intravenously to the rats.The kidneys harvested 6 h later were transplanted to Lewis recipients.Cyclosporine was intromuscularly given daily to the recipient rats for 10 days.Right kidneys were resected from recipients on day 10.Creatinine level was examined on day 14,21,28,and 35.Renal allografts harvested on day 35 were pathologically detected.The irnmunochemistry expression of interleukin (IL)-1β and tumor necrotic factor (TNF)-α in renal allograft tissue was tested.Result Serum creatinine levels in G2 were remarkably higher than those in G1 and G3 (P＜0.01) on day 14,21,28,and 35 postoperatively.The creatinine levels on the above mentioned time points had no statistically significant difference between G3 and G1 except on day 21.Postoperative pathological changes in G2 of both pronounced infiltration of mononuclear cells and tubular epithelia[inflammation were notably increased in renal allografts as compared with those in G1 and G3.There was no obvious difference between G1 and G3 in infiltrated mononuclear cells and tubular epithelial inflammation.Positive expression levels of both IL-1β and TNF-α in glomerular,tubular and interstitial epithelial cells were statistically enhanced in G2 as compared with those in G1 and G3 (H =7.210,P =0.027),while there was no statistically significant difference in the expression of both IL-1[β and TNF-α between G1 and G3.Conclusion Brain dead donor rats pretreated with bone marrow MSCs might reduce renal allograft injury via decreasing both inflammatory cell infiltration and IL-1β and TNF-α expression.

AIM:To investigate the effects of ethane 1,2-dimethanesulfonate (EDS) preconditioning on renal ischemia/reperfusion (I/R) injury in male Sprague-Dawley (SD) rats.METHODS: Male SD rats (n=48) were ran-domly assigned to 6 groups:blank, sham, I/R, EDS+I/R, EDS+testosterone (TST) +I/R, and castration (Cast)+I/R.The renal pedicles were bilaterally occluded with a microvascular clamp for 45 min to establish renal I/R-induced in-jury model.Bilateral orchiectomy was conducted 2 weeks before surgery .EDS (75 mg/kg) was intraperitoneally injected 5 d before operation .Blood samples were collected 24 h after reperfusion from the vena orbitalis posterior plexus .Luteinizing hormone (LH), TST, serum creatinine (SCr), blood urea nitrogen (BUN), and kidney injury molecule-1 (KIM-1) were detected.The renal tissues were harvested to measure the level of TNF-αand the expression of Fas mRNA and caspase-3 protein.RESULTS:Serum TST levels in EDS +I/R group and Cast +I/R group were below the minimum detectable threshold.Compared with other groups , the rats in EDS+I/R group and Cast +I/R group had higher levels of SCr , BUN and KIM-1 (P<0.05).SCr and BUN levels showed no significant difference between EDS +I/R group and Cast +I/R group (P>0.05), but KIM-1 level in EDS+I/R group was lower than that in Cast +I/R group (P<0.05).After reper-fusion for 24 h, the levels of TST and LH in EDS +I/R group, Cast+I/R group and EDS+TST+I/R group were lower than those 1 h before operation (P<0.05).Compared with Cast+I/R and I/R group, the TNF-αlevel and expression of Fas mRNA and caspase-3 protein were significantly decreased in EDS +I/R group ( P<0.05 ) .CONCLUSION: EDS preconditioning substantially reduces the serum TST level , thus attenuating I/R-induced acute renal injury .TNF-α-induced Fas/FasL pathway may be involved in this process .

Nowadays, there is still no mature gene delivery system for safe and effective transfection on primary dendritic cells (DC). Herein, we constructed a liposome-based gene delivery system for primary DCs and optimized the preparation method to improve the transfection efficiency of siRNA on primary DCs. In this study, different methods, including co-incubation method, ethanol injection method, and protamine compound method, were used to prepare liposome/siRNA complexes based on different cationic lipids. Moreover, particle size, zeta potential, siRNA loading capacity, safety, stability, uptake efficiency and gene silencing efficiency of various liposome/siRNA complexes were detected to screen the optimal cationic lipid as well as its preparation method. We demonstrated that the OA2/siRNA delivery system prepared by the co-incubation method exhibited the best safety, uptake efficiency and gene silencing effect, compared to other siRNA delivery systems including the commercial Lipo2000. In summary, we provide a safe and effective gene delivery vector for primary DC cells through simple preparation method, which could also offer a gene delivery platform for other immune cells.