A new sesquiterpenoid, 8α-hydroxy-6β-methoxy-1-oxoeremophila-7 (11), 9 (10) -diene-12, 8-olide (1) and five known compounds, petasin (2), caffeic acid (3), hepta-cosanol (4), β-sitosterol (5) and β-daucosterol (6) have been isolated from the roots of Ligularia intermedia. The compounds were isolated by column chromatography on silica gel and Sephadex LH-20, and identified based on spectral analyses (MS, 1H-NMR, 13C-NMR).
Asteraceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Magnetic Resonance Imaging ; Molecular Structure ; Plant Roots ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Purpose The aim of this study was to investigate the relationship between autophagy gene Beclin1 and immune response effector classical HLA Ⅰ,Ⅱ in SKOV3 cells.To explore the role of Beclin1 in immunity in ovarian cancer cells which were transfected with the vector of Beclin1.Methods RT-PCR and Western blot were used to detect the expression of Beclin1 and HLA Ⅰ,Ⅱ in SKOV3 cells.Fluorescence microscope was carried out to observe the unique autophagosome in SKOV3 cells.MTT was used to analyze the proliferation of the Beclin1 over-expressed SKOV3 cells.Results Transfection SKOV3 cells with Beclin1 vector could induce Beclin1 transcription and translation approximately 5 and 2 times compared with empty vector group respectively.The autophagosome stained by MDC was observed by fluorescence microscope.And much more green fluorescence signal was observed in Beclin1 vector group.RT-PCR and Western blot indicated that HLA Ⅰ,Ⅱ induced by transfection with extrinsic Beclin1.The allelic transcriptions of HLA Ⅰ-A,B,C and HLA Ⅱ-DP,DQ,DR in extrinsic Beclin1 group were approximately 2,1.6,3 and 2,6,3 times compared with empty vetcor group or untreated group,respectively.The results of Western blot showed that HLA Ⅰ,Ⅱ in Beclin1 vector group induced as much as 2 and 1.6 times compared with empty vetcor group or untreated group,respectively.The results of MTT showed that the proliferation of SKOV3 cells treated with Beclin1 vector was significantly suppressed.The percentage of suppression in Beclin1 vector group at 24 h,48 h,72 h and 96 his42.6％,37.8％,24.35％,14.81％ compared with untreated group or empty vector group respectively.Conclusion The enhancement of autophagy by over-expression of Beclin1 could induce HLA Ⅰ,Ⅱ transcription and translation in SKOV3 cells.The expression of HLA Ⅰ,Ⅱ may be responsible for triggering the immune response in ovarian cancer.Over-expression of Beclin1 could inhibit the proliferation of SKOV3 cells which were transfected with extrinsic Beclin1.

Etanercept has been shown to be effective for the treatment of moderate-to-severe plaque psoriasis.Since most clinical trials examined etanercept in combination with other drugs,the efficacy and safety of etanercept monotherapy for moderate-to-severe plaque psoriasis have not been well established.This prospective study enrolled 61 Chinese patients with moderate-to-severe plaque psoriasis to explore the efficacy and safety of etanercept monotherapy.These patients were treated with etanercept at a subcutaneous dose of 25 mg,twice a week,for 12 weeks.All the 61 patients completed the treatment and showed significant improvement in psoriasis area and severity index (PASI) scores.At 4,8,and 12 weeks after treatment,the response rates (PASI75) were 0％,21.31％,and 40.98％,respectively.It was concluded that etanercept monotherapy is efficacious and safe for patients with moderate-to-severe plaque psoriasis.

BACKGROUNDMany treatment options for lower limb ischemia are difficult to apply for the patients with poor arterial outflow or with poor general conditions. The effect of medical treatment alone is far from ideal, especially in patients with diabetic foot. A high level amputation is inevitable in these patients. This study aimed to explore the effect of transplantation of autologous bone marrow mononuclear cells on the treatment of lower limb ischemia and to compare the effect of intra-arterial transplantation with that of intra-muscular transplantation.

METHODSIn this clinical trial, 32 patients with lower limb ischemia were divided into two groups. Group 1 (16 patients with 18 affected limbs) received transplantation of autologous bone marrow mononuclear cells by intra-muscular injection into the affected limbs; and group 2 (16 patients with 17 affected limbs) received transplantation of autologous bone marrow mononuclear cells by intra-arterial injection into the affected limbs. Rest pain, coldness, ankle/brachial index (ABI), claudication, transcutaneous oxygen pressure (tcPO(2)) and angiography (15 limbs of 14 patients) were evaluated before and after the mononuclear cell transplantation to determine the effect of the treatment.

RESULTSTwo patients died from heart failure. The improvement of rest pain was seen in 76.5% (13/17) of group 1 and 93.3% (14/15) of group 2. The improvement of coldness was 100% in both groups. The increase of ABI was 44.4% (8/18) in group 1 and 41.2% (7/17) in group 2. The value of tcPO(2) increased to 20 mmHg or more in 20 limbs. Nine of 15 limbs which underwent angiography showed rich collaterals. Limb salvage rate was 83.3% (15/18) in group 1 and 94.1% (16/17) in group 2. There was no statistically significant difference in the effectiveness of the treatment between the two groups.

CONCLUSIONSTransplantation of autologous bone marrow mononuclear cells is a simple, safe and effective method for the treatment of lower limb ischemia, and the two approaches for the implantation, intra-muscular injection and intra-arterial injection, show similar results.

Aged ; Aged, 80 and over ; Blood Gas Monitoring, Transcutaneous ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; Female ; Humans ; Ischemia ; therapy ; Leg ; blood supply ; Leukocytes, Mononuclear ; transplantation ; Male ; Middle Aged ; Transplantation, Autologous

Objective: To prepare Juglone-loaded poly lactic-co-glycolic acid nanoparticles (Jug-PLGA-NPs), and investigate their physicochemical properties, release characteristics in vitro and anti-tumor activities on A375 melanoma cells in vitro. Method: Jug-PLGA-NPs were prepared by emulsification-solvent evaporation method. Then the particle size, encapsulation efficiency, drug loading rate and in vitro release characteristics were investigated. Fluorescence microscopy was used to observe the uptake of PLGA-NPs in vitro. The distribution of PLGA-NPs in BALB/c nude mice after tail vein injection was observed by the small living animal imaging system. Their inhibition effect on proliferation of A375 cells was detected by thiazolyl blue tetrazolium bromide (MTT) assay. Apoptosis rate and cell cycle detection were performed by flow cytometry. Western blot was used to determine the protein kinase B (Akt), phosphorylated Akt (p-Akt) and cyclinD₁. Result: The average particle size of the prepared Jug-PLGA-NPs was (149.6±21.5) nm, entrapment rate of (68.39±2.51)%, and drug-loading rate of (5.07±0.98)%, showing good sustained-release characteristics. PLGA-NPs showed good penetration and targeting properties in cellular uptake in vitro and in vivo imaging. Different concentrations of Jug-PLGA-NPs could significantly inhibit the proliferation and promote apoptosis of A375 cells in a time and concentration dependent manner (P<0.05), and its 48 h effect was superior to that of the same concentration of Jug. The mechanism may be related to the regulation of Akt phosphorylation level, down-regulation of cyclinD₁ expression (P<0.05) and the cell-cycle arrest in G₀/G₁ phase (P<0.05). Conclusion: The Jug-PLGA-NPs are easy to prepare and have good sustained-release characteristics, tumor targeting and anti-tumor ability, providing a new pharmaceutical dosage form for the future clinical application of Jug.

OBJECTIVE: To compare the clinical efficacy between moxibustion and acupuncture for knee osteoarthritis (KOA), and to observe the effect on serum tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), superoxide dismutase (SOD) and malondialdehyde (MDA). METHODS: A total of 60 patients with KOA were randomized into a moxibustion group (30 cases, 2 cases dropped off) and an acupuncture group (30 cases, 2 cases dropped off). In the aucpuncture group, acupuncture was applied at Neixiyan (EX-LE 4), Dubi (ST 35), Heding (EX-LE 2), Xuehai (SP 10), Liangqiu (ST 34), Zusanli (ST 36) and point on the affected side for 30 min.In the moxibustion group, moxibustion was adopted at knee for 60 min. The treatment was given once every two days for 4 weeks, totally 14 times. Before and after treatment, the western ontario and McMaster universities osteoarthritis index (WOMAC) score was compared, and the therapeutic effect was evaluated in the two groups. The contents of serum TNF-αand IL-1β, the activity of serum SOD and the serum level of MDA were detected in the two groups. RESULTS: Compared before treatment, the WOMAC scores and the contents of serum TNF-α, IL-1β and MDA after treatment were reduced (<0.05), the activity of serum SOD was increased (<0.05) in the two groups. In the moxibustion group, the WOMAC score and the contents of serum TNF-α, IL-1β and MDA after treatment were lower than the acupuncture group (<0.05), the activity of serum SOD was higher than the acupuncture group (<0.05). The total effective rate was 89.3% (25/28) in the moxibustion group, which was superior to 42.9% (12/28) in the acupuncture group (<0.05). CONCLUSION Moxibustion and acupuncture can relieve KOA symptoms, and the therapeutic effect of moxibustion is superior to acupuncture. The mechanism may be related to the reduction of serum inflammatory factor and oxidative stress factor.

OBJECTIVE: To establish a solvent desorption-gas chromatography method for determination of methyl pentane in workplace air. METHODS: Methyl pentane in workplace air was collected with activated carbon tube and desorbed with carbon disulfide, then separated by DB-1 capillary column, detected by flame ionization detector, and quantified by standard curve method. RESULTS: Good linearity was obtained in the range of 1.98-6 600.00 mg/L with the correlation coefficient of 0.999 9. The minimum detection limit and the minimum quantification limit were 0.06 and 0.20 mg/L, respectively. The minimum detection concentration and minimum quantification concentration was 0.04 and 0.14 mg/m~3, respectively(calculated by collecting 1.5 L of air sample). The average desorption efficiency was 97.3%-102.2%. The within-run relative standard deviation(RSD) and the between-run RSD were 0.4%-0.9% and 0.3%-3.0%, respectively. The sampling efficiency was 96.7%-100.0% and the penetration capacity was 8.68 mg. The samples can be stored at room temperature for at least 14 days. CONCLUSION: This method is suitable for methyl pentane detection in workplace air.

BACKGROUNDDendritic cells are professional antigen-presenting cells found in an immature state in epithelia and interstitial space, where they capture antigens such as pathogens or damaged tissue. Matrix metallopeptidase 13 (MMP-13), a member of the collagenase subfamily, is involved in many different cellular processes and is expressed in murine bone marrow-derived dendritic cells (DCs). The function of MMP-13 in DCs is not well understood. Here, we investigated the effect of MMP-13 on DC maturation, apoptosis, and phagocytosis.

METHODSBone marrow-derived dendritic cells were obtained from C57BL/6 mice. One short-interfering RNA specific for MMP-13 was used to transfect DCs. MMP-13-silenced DCs and control DCs were prepared, and apoptosis was measured using real-time polymerase chain reaction and Western blotting. MMP-13-silenced DCs and control DCs were analyzed for surface expression of CD80 and CD86 and phagocytosis capability using flow cytometry.

RESULTSCompared to the control DCs, MMP-13-silenced DCs increased expression of anti-apoptosis-related genes, BAG1 (control group vs. MMP-13-silenced group: 4.08 ± 0.60 vs. 6.11 ± 0.87, P = 0.008), BCL-2 (control group vs. MMP-13-silenced group: 7.54 ± 0.76 vs. 9.54 ± 1.29, P = 0.036), and TP73 (control group vs. MMP-13-silenced group: 4.33 ± 0.29 vs. 5.60 ± 0.32, P = 0.001) and decreased apoptosis-related genes, CASP1 (control group vs. MMP-13-silenced group: 3.79 ± 0.67 vs. 2.54 ± 0.39, P = 0.019), LTBR (control group vs. MMP-13-silenced group: 9.23 ± 1.25 vs. 6.24 ± 1.15, P = 0.012), and CASP4 (control group vs. MMP-13-silenced group: 2.07 ± 0.56 vs. 0.35 ± 0.35, P = 0.002). Protein levels confirmed the same expression pattern. MMP-13-silenced groups decreased expression of CD86 on DCs; however, there was no statistical difference in CD80 surface expression. Furthermore, MMP-13-silenced groups exhibited weaker phagocytosis capability.

CONCLUSIONThese results indicate that MMP-13 inhibition dampens DC maturation, apoptosis, and phagocytosis.

Animals ; Apoptosis ; drug effects ; physiology ; Bone Marrow Cells ; cytology ; Dendritic Cells ; cytology ; drug effects ; metabolism ; Female ; Lipopolysaccharides ; pharmacology ; Matrix Metalloproteinase 13 ; metabolism ; physiology ; Mice ; Mice, Inbred C57BL ; RNA, Small Interfering

Aim To study the neuroprotective effect of rhubarb extract on MCAO model rats and explore its mechanism of action. Methods Forty-five SPF male Sprague-Dawley rats were randomly divided into sham group, MCAO group, and MCAO + rhubarb group. MCAO model was prepared by silk plug method, and rhubarb extract was administered at a concentration of 200 mg · kg

OBJECTIVE: To verrify the anti-tumor efficacy and toxicity between juglone (Jug) and Jug-loaded PLGA nanoparticles (Jug-PLGA-NPs). METHODS: Jug-PLGA-NPs were prepared by ultrasonic emulsification. The anti-tumor activity of Jug (2, 3, 4 µg/mL) and Jug-PLGA-NPs (Jug: 2, 3, 4 µg/mL) in vitro was measured by MTT assay and cell apoptosis analysis. The distribution, anti-tumor effect and biological safety in vivo was evaluated on A375 nude mice. RESULTS: With the advantage of good penetration and targeting properties, Jug-PLGA-NPs significantly inhibited proliferation and migration of melanoma cells both in vitro and in vivo (P<0.05 or P<0.01) with acceptable biocompatibility. CONCLUSIONS Jug can inhibit the growth of melanoma but is highly toxic. With the advantage of sustained release, tumor targeting, anti-tumor activity and acceptable biological safety, Jug-PLGA-NPs provide a new pharmaceutical form for future application of Jug.
Animals ; Cell Line, Tumor ; Delayed-Action Preparations/therapeutic use* ; Drug Carriers/therapeutic use* ; Melanoma/pathology* ; Mice ; Mice, Nude ; Nanoparticles ; Naphthoquinones ; Particle Size ; Polylactic Acid-Polyglycolic Acid Copolymer/therapeutic use*