Objective To explore the effect of suramin on the epithelial-mesenchymal transition (EMT) and the excretion of transforming growth factor-β1 (TGF-β1) in peritoneal mesothelial cells (PMCs) induced by high concentrations of glucose solution (GS).Methods Cultured PMCs were divided into three groups:(1) normal control group; (2) GS-treated group:cells were treated with 1.5％,2.5％,4.25％ GS for 12 h,24 h,48 h,respectively; (3) Suramin-treated group:PMCs cultured with 4.25％ GS were exposed to different doses of suramin (25,50,100 μmol/L) for 48 h.Expression levels of α-smooth muscle actin (α-SMA) and E-cadherin were detected by Western blotting and the concentration of TGF-β1 in the culture supernatant was determined by ELISA.Results Compared with normal control group,GS-treated PMCs exhibited a time-dependent increase in the expression of α-SMA,and decrease in the expression of E-cadherin.GS also stimulated PMCs to secrete TGF-β1.In the presence of suramin,GS-induced α-SMA expression and TGF-β1 production were reduced,E-cadherin expression was increased.Conclusions Suramin can inhibit high glucose-induced EMT of PMCs by down-regulating the expression of TGF-β1.Suramin may be a novel therapeutic agent for the treatment of peritoneal fibrosis.

Objective The aim of this study was to investigate the effect of hepatitis C virus (HCV) replication on expression of silent information regulator 1 (SIRT1) and glucose metabolism of hepatocytes using Huh 7.5 cells harboring HCV replicon.Methods The level of reactive oxygen species (ROS),value of nicotinamide adenine dinucleotide (NAD+)/reduced form of nicotinamide adenine dinucleotide (NADH) was detected by flow cytometry and chromatometry.The activity,mRNA expression,and protein level of SIRT1 were detected by a scintillation counter,real-time fluorescence quantitative polymerase chain reaction (RT-PCR),and Western blot,respectively.Glucose uptake by hepatocytes and gluconeogenesis were detected using radioactive isotope method and glucose oxidase method.The mRNA levels of SIRT1 downstream glucose-metabolism genes were measured by RT-PCR.Measurement date were compared by t test.Results In replicon cells,the level of ROS (3.8±0.5 vs 1.0±0.2; t=12.736,P＜0.01) was increased and the value of NAD+/NADH (0.03±0.01 vs 0.12±0.03; t=6.971,P＜0.01) decreased compared with Huh 7.5 cells.The activity (0.3±0.1 vs 1.0±0.2; t=7.668,P＜0.01),mRNA expression(0.4±0.1 vs 1.0± 0.3; t=4.648,P＜0.01) and protein level(0.3±0.1 vs 0.8±0.2; t=5.941,P＜0.01) of SIRT1 were reduced.Inhibition of SIRT1 not only increased insulin receptor substrate-1 (IRS-1) phosphorylation (0.7±0.2 vs 0.4±0.1; t=3.286,P＜0.01),decreased protein kinase B (Akt) phosphorylation (0.3 ± 0.1 vs 0.6 ± 0.2; t=3.286,P＜0.01),down regulated cell surface expression of glucose transporler 2 (GLUT2,0.4±0.1 vs 1.0 ± 0.2; t =6.573,P＜0.01) and suppressed cellular glucose uptake (count per minute:4600±500 vs 21 000±4600; t=8.682,P＜0.01); but also decreased phosphorylation of forkhead box O1 (FoxO1,0.2＝0.1 vs 0.5±0.1; t=5.196,P＜ 0.01),up-regulated phosphoenolpyruvate carboxykinase (PEPCK,2.8±0.6 vs 1.0±0.3; t=6.573,P＜0.01) and glucose 6-phosphatase (2.6±0.5 vs 1.0±0.2; t=7.278,P＜0.01) genes,and promoted glucose production (2.5±0.5 vs 1.0±0.2; t=5.543,P＜0.01).Conclusions HCV replication decreases NAD+/NADH ratio,which might down-regulate the activity and the expression of SIRT1,leading to changes in the expression profile of glucose metabolism related genes and causing glucose metabolism disorders of hepatocytes by a decrease in glucose uptake and an increase in glucose production,and promotes HCV replication.

Matrix_assisted laser desorption ionization_time of flight tandem mass spectrometry ( MALDI_TOF/TOF MS) and electrospray ionization_quadrupole_time of flight mass spectrometry ( ESI_Q_TOF MS) were used to confirm the structure of cyclic lipopeptide daptomycin fastly. First, the relative molecular weight 1916. 7107 of daptomycin was measured by ESI with error 0. 0007. The sample’s doubly charged peak m/z 809. 848 was selected as precursor ion for ESI_MS/MS analysis, and the exocyclic amino acid sequence C9 H19 CO_Trp_Asn_Asp was successfully matched. Second, the experimental conditions of cleaving daptomycin by lithium hydroxide ( LiOH) were optimized and the ring_opened process was monitored by MALDI_TOF/TOF MS. After obtaining ring_opened product with purity of above 95%, the MS/MS measurements by MALDI and ESI were carried out. The b+and y+of ring_opened product were completely matched, which confirmed the amino acid sequence of daptomycin. Finally, ESI_MS/MS conditions of ring_opened product were further optimized to obtain more low mass fragment ions for analyzing the structure of fatty acid chain and the cleavage pattern of fat chain in mass spectrometry was proposed. The method was fast, convenient, accurate and reliable for identifying cyclic lipopeptide compounds.

Objective To develop a method for the determination of five furostanol saponins(timosaponin N,timosaponin L, timosaponin BⅡ,25R-timosaponin BⅡ,and 25S-officinalisnin-Ⅰ)in rhizome and fibrous root of Anemarrhena asphodeloides Bge. by HPLC with the charged aerosol detector(CAD). Methods The analysis was performed on TechMate C18-ST-II(250 mm×4.6 mm,5μm)with acetonitrile:water(22:78,V/V),the flow rate of 1.0 ml/min and column temperature at 30℃. The Corona parameters were as follows:sampling rate 10 Hz,filter 5 s,and the nebulizer temperature 55℃. Results The approach showed good linearity for five saponins. The correlation coefficients(r2)for calibration curves varied from 0.9992 to 0.9998. The limits of detection(LOD)were 0.28,0.92,0.92,0.92 and 0.92 ng for five steroidal saponins,respectively. The limits of quantitation(LOQ)were found to be 0.92, 2.77,2.77,2.77 and 2.76 ng,respectively. RSD calculated from peak area of precision,repeatability and stability in 48 h were all less than 3.0%. The average recoveries of timosaponin N,timosaponin L,timosaponin BⅡ,25R-timosaponinBⅡ,and 25S-officinalis-nin-Ⅰwere 98.17%,101.37%,98.53%,97.63%,and 98.17%,respectively. Conclusion The developed method is accurate,reli-able,which could be applied to the quality control of multiple components in A. asphodeloides Bge.

Objective To design and synthesize compounds with protein tyrosine kinase(PTK)inhibitory activity with L029 as the lead compound. Methods L029 derivatives were designed and synthesized from L029 by reduction and/or substitution with the 3-dimethylamino-1-propyl,methyl acetate,methyl propionate in its active H and other sites. PTK activity was measured by enzyme-linked immunosorbent assay(ELISA). The inhibitory rate was calculated to screen out the compounds with PTK inhibitory activity. Re-sults Five target compounds were synthesized and their structures were confirmed by 1H NMR and MS. Three compounds T2,T3 and T5 were screened out with strong PTK inhibitory activity. Conclusion The synthetic routes of the target compounds are simple with mild reaction condition,and 3 compounds show strong inhibitory activity by ELISA. These results can provide reference for the further design and synthesis of this kind of molecules.

Objective To design and synthesize novel 2-indolone derivatives as the c-Met kinase inhibitors. Methods With c-Met kinase inhibitor SU11274 as lead compound,a series of 2-indolone derivatives were designed according to the concept of bioiso-sterism. Then the target compounds(10a-10r)were synthesized from 2-indolone through 5-chlorosulfonation with chlorosulfonic acid, sulfonamidation with intermediate 3,condensation with 6a-6h,7a-7h and 4a-4b,respectively. Their inhibitory activity against c-Met and proliferation of MCF-7 cells were evaluated. Results and Conclusion The designed compounds were successfully prepared and their structures were confirmed by 1H NMR and ESI-MS. Some compounds had certain inhibitory activity against c-Met and prolif-eration of MCF-7 cells. An initial structure-activity relationship analysis of these compounds was performed to provide useful informa-tion for further optimization of their structures.

Objective To investigate the feasibility and short-term efficacy of endplate reduction percutaneous pedicle screw (ERPPS) technique combined with short-segment percutaneous pedicle screw fixation for the treatment of AO type A3 thoracolumbar fractures.Methods A retrospective case control study was conducted to analyze the clinical data of 36 patients with type A3 thoracolumbar fractures without neurological symptoms and with comminuted endplates admitted to 903 Hospital of PLA from December 2015 to January 2018.Fifteen patients (Group A) were treated with ERPPS technique combined with short-segment percutaneous pedicle screw fixation,including 11 males and four females,aged (37.9 ±8.3)years.The injured segments were at T11 in 1 patient,T12 in 3,L1 in 6,L2 in 3 and L3 in 2.Simple short-segment percutaneous pedicle screw reduction and internal fixation was performed in 21 patients (Group B),including 14 males and seven females,aged (37.3 ± 9.5)years.The injured segments were at T~ in two patients,T12 in six,L1 in seven,L2 in four and L3 in two patients.The operation time,intraoperative bleeding and complications were recorded.The anterior vertebral body height ratioin (AVBHr),middle vertebral body height ratio (MVBHr),posterior vertebral body height ratio (PVBHr),Cobb angle of kyphosis and wedge angle of injured vertebrae were calculated based on the measurement by X-ray films taken before operation,during operation (after regular reduction),3 days after operation and 6 months after operation.Visual analogue scale (VAS) and Oswestry dysfunction index (ODI) were used to assess the pain and functional improvement.Results All patients were followed up for 11-30 months [(19.1 ± 5.0) months].The operation time was (62.8 ± 4.4)minutes in Group A and (60.1 ± 4.7)minutes in Group B (P ＞ 0.05).The intraoperative blood loss was (48.5 ± 5.1) ml in Group A and (48.0 ± 4.9) ml in Group B (P ＞ 0.05).All the incisions were healed by first intention without complications.The MVBHr of injured vertebra was (84.8 ± 4.4) ％ in Group A and (68.1 ±8.8)％ in Group B (P＜0.05).The MVBHr 6 months after operation was (81.3 ±4.9)％in Group A,significantly better than that in Group B [(63.6 ± 8.1) ％] (P ＜ 0.05).At 6 months after surgery,the kyphosis Cobb angle [(11.3 ± 3.2) °],the wedge angle [(10.5 ± 2.1) °] of the injured vertebra and the VAS [(1.1 ± 0.7) points] of Group A were significantly better than those of Group B [(13.4±2.3)°,(12.1 ±2.2) °and (1.9±1.1)points] (P＜0.05).There were no significant differences in AVBHr,PVBHr and ODI between the two groups (P ＞ 0.05).Conclusion For type A3 thoracolumbar fractures with endplate comminuted injury and without neurological symptoms,the ERPPS technique can effectively reduce the collapse of the central part of the upper endplate and improve the clinical results (less reduction loss and back pain) after short-segment percutaneous pedicle screw reduction and internal fixation under the premise of strict indications.

Membranous nephropathy (MN) is a type of glomerular disease characterized by diffuse thickening of glomerular basement membrane with subepithelial immune complex deposition, and traditional diagnosis of MN mainly relies on the pathological results of renal biopsy. In recent years, the emergence of biomarkers related to MN such as phospholipase A2 receptor and thrombospondin type-1 domain-containing 7A has changed the diagnosis and treatment mode of MN, providing a new basis for the diagnosis, treatment and prognosis of MN. MN patients with positive specific target antigens exhibit different clinical manifestations and prognoses. Specific target antigens can not only guide diagnosis, but also has predictive value for prognosis. Immunosuppressive therapy is a common treatment for idiopathic MN patients, and the emergence of novel medications such as biologics represents a advance in the treatment of MN, providing a broader array of options for managing the condition. Conversely, the treatment approach for secondary MN primarily targets the management of the primary disease. Based on multiple and new literature, we reviewed the researches progress of target antigens and immunosuppressive therapy related to MN, so as to provide references for clinical diagnosis and treatment of MN.

The blood-brain barrier (BBB) constitutes a major obstacle to effective delivery of drugs to the brain. Recent technological advances have led to improvements in brain-targeted drug delivery. In this review, we summarize existing technologies for efficient drug delivery across the BBB. We discuss the mecha-nisms of current BBB-based drug delivery strategies and introduce prospects for application in brain-targeted delivery of traditional Chinese medicine. We highlight the use of physical techniques for brain-targeted drug delivery, including electroporation, ultrasound, magnetophoresis, microneedles, microwaves, and laser. The characteristics of these techniques and relevant studies employing these approaches are discussed. In general, microneedles, lasers, ultrasound, electroporation, magnetophoresis, and microwaves are effective for drug delivery across the BBB. Notably, the synergistic effects of multiple approaches are superior to the additive effects of each technique in isolation. Our review provides guidance for the practical application of brain-targeted drug delivery techniques in an efficient and safe manner.