1.Preparation of cell penetrating peptide TAT and cleavable PEGco-modified liposomes loaded with paclitaxel and its in vitro apoptosis assay.
Han FU ; Guan-Lian HU ; Qin HE
Acta Pharmaceutica Sinica 2014;49(7):1054-1061
The preparation method, serum stability, efficiency of cellular uptake and apoptosis induction of the cell penetrating peptide TAT and cleavable PEG co-modified liposomes loaded with paclitaxel (C-TAT-Lipo) were investigated. The best preparation procedure was performed by orthogonal test based on single factor screening method. First, the paclitaxel (PTX)-loaded liposomes were prepared by filming-rehydration method, evaluated with entrapment efficiency and polydispersity index. The morphology of C-TAT-Lipo was characterized by transmission electron microscopy. Turbidity variations were monitored in the presence of fetal bovine serum (FBS) to evaluate the serum stability of the liposomes developed here. Next, the efficiency of cellular uptake of different Rho-PE-labeled liposomes on B16F1 cells in vitro was evaluated by confocal laser scanning microscopy (CLSM) and flow cytometry. The quantitative analysis of apoptosis induced by different PTX-loaded liposomes was performed by Annexin V-FITC/PI double staining. The optimal formulation was as follows: Chol : lipid: 1 : 8 (molar ratio); drug : lipid: 1 : 40 (mass ratio); lipid concentration: 3 mmol x L(-1); temperature of hydration: 25 degrees C. The mean size and polydispersity index of C-TAT-Lipo were about (97.97 +/- 3.68) nm and 0.196 +/- 0.037, the zeta potential was (-0.89 +/- 0.45) mV, the entrapment efficiency of paclitaxel was (90.16 +/- 1.53)%. The particle sizes did not exhibit significant variations in 50% FBS over 24 h at 37 degrees C. The efficiency of cellular uptake of the C-TAT-Lipo increased 1.40 fold following the cleavage of PEG. Apoptosis analysis showed 59.3% increase of the apoptosis and necrosis profile of C-TAT-Lipo after the detachment of PEG shells, which was markedly higher than that of N-TAT-LP with or without glutathione and SL, respectively. The results indicate that the C-TAT-Lipo is successfully prepared by filming-rehydration method and shows significant antitumor activities.
Animals
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Annexin A5
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Apoptosis
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Cell Line, Tumor
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Cell-Penetrating Peptides
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pharmacology
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Fluorescein-5-isothiocyanate
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analogs & derivatives
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Liposomes
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chemistry
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Melanoma, Experimental
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Mice
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Microscopy, Confocal
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Paclitaxel
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pharmacology
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Particle Size
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Polyethylene Glycols
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chemistry
2.Influence of Shenqifuzheng injection on efficacy of chemotherapy and immune function in patients with gas-tric cancer after surgery
Yufen XIE ; Ruiyang FU ; Xinjun GUAN
Chinese Journal of Primary Medicine and Pharmacy 2015;(20):3078-3081
Objective To explore the influence of Shenqifuzheng injection on efficacy of chemotherapy and immune function in patients with gastric cancer after surgery.Methods 60 postoperative patients with gastric cancer were selected as the research subjects,and were randomly divided into the control group and observation group by the digital table,30 cases in each group.The control group was given FOLFOX regimen chemotherapy.The observation group received FOLFOX regimen combined with the treatment of Shenqifuzheng injection.The recurrence and metasta-sis,survival,immune function and adverse reactions of the patients were observed and the patients were observed before and after chemotherapy.Results The chemotherapy cycle of the observation group was (3.95 ±0.40),which was significantly higher than (3.40 ±0.30)of the control group,the difference was statistically significant (t =6.025,P <0.01).The progression free survival in the observation group was (9 ±2.4)months,which was significant-ly longer than (6.6 ±3.2)months in the control group,the difference was statistically significant (t =3.286,P <0.01).The reduction rate in the observation group was 6.67%,which was lower than 26.67% in the control group, the difference was statistically significant (χ2 =4.320,P <0.05).The incidence rate of nausea and vomiting in the observation group was 6.67%,which was lower than 26.67% in the control group,the difference was statistically sig-nificant (χ2 =4.320,P <0.05).KPS[(83.51 ±15.37)points]and QLQ -C30[(82.01 ±9.45)points]of the observation group were significantly higher than those in the control group [KPS (74.62 ±13.22)points and QLQ -C30 (74.62 ±13.22)points],the differences were statistically significant (t =2.298,2.491,all P <0.05).After chemotherapy,gastric cancer patients ratio of CD +3 ,CD +4 ,CD +4 /CD +8 in the observation group were higher than those of the control group and the ratio of CD +4 /CD +25 in the observation group was lower than control group,the differences were statistically significant (all P <0.05).Conclusion Shenqifuzheng injection combined with FOLFOX chemo-therapy for gastric cancer patients has low adverse reaction rate,and can effectively improve the immune function.
3.Study on Inhabition Effect of Chitlsan Oligosaccharide on Tumour
Jie GUAN ; Dianjun LI ; Zhenkun FU
Journal of Medical Research 2006;0(01):-
Objective To observe the inhibition effect of chitlsan oligosaccharide on tumour. Methods The growth of tumour were observed after chitlsan oligosaccharide was injected into abdominal cavity(ip) or introtumour of mice;And the anti proliferation activities of chitlsan oligosaccharide on cells were evaluated by means of MTT assay. Results Growth of tumour were inhibited by chitlsan oligosaccharide, necrosis was observed in the tumour tissue.Chitlsan oligosaccharide affected the growth of cell. The effect was related to concentration of chitlsan oligosaccharide,but not depending of culture time.The apoptosis of cells could be seen under the transmission electron.Conclusion Chitlsan oligosaccharide could inhibit the growth of tumour and might induce apoptosis of cell.
4.Determination of Tanshinone Ⅱ_A in Huganning Tablets by HPLC
Xiaoliu FU ; Lei GUAN ; Demin ZHANG
China Pharmacy 2007;0(30):-
OBJECTIVE:To establish an HPLC method for the determination of Tanshinone Ⅱ A in Huganning tablets.METHODS:The determination was performed by HPLC using C18 column(200 mm?4.6 mm,5 ?m) with the column temperature maintained at 25 ℃.The mobile phase consisted of methanol-water(80∶20) at a flow rate of 1.0 mL?min-1 and a UV detection wavelength of 270 nm.RESULTS: The calibration curve of Tanshinone Ⅱ A was in good linearity over the range of 4.999~44.999 ?g?mL-1(r=0.999 7) and the mean recovery was 99.07%(RSD=0.33%,n=9).CONCLUSION: The method is sensitive,accurate and reproducible,and it can be used for the quality control of Huganning tablets.
5.Impacts on analgesia and detumescence in ankle sprain treated with acupuncture at Xiaojie point combined with tendon-regulation manipulation.
Wei-Bin DU ; Guan-Ai BAO ; Ren-Fu QUAN
Chinese Acupuncture & Moxibustion 2014;34(7):647-650
OBJECTIVETo compare the efficacy difference of analgesia and detumescence on ankle sprain among acupuncture at Xiaojie point combined with tendon regulation manipulation, acupuncture at Xiaojie point and tendon regulation manipulation.
METHODSSixty cases of ankle sprain were randomized into a combined therapy group, a Xiaojie point group and a tendon-regulation manipulation group, 20 cases in each one. The combined therapy of acupuncture at Xiaojie point and tendon regulation manipulation, the acupuncture at Xiaojie point and the simple application of tendon-regulation manipulation were applied respectively in the three groups, once a day, 3 treatments were required. The symptom score such as pain, ecchymosis, swelling and motor dysfunction and the total score were observed before and after treatment in the three groups. The different values of pain and swelling scores were compared before and after treatment in the three groups. The efficacy was compared among the groups.
RESULTSThe total effective rate was 100.0% (20/20) in each group. But the curative rate was 85.0% (17/20) in the combined group, 65.0% (13/20) in the Xiaojie point tion manipulation group. After treatment, the symptom scores of pain, ecchymosis, swelling and motor dysfunction and the total score were all improved as compared with those before treatment in the three groups (P < 0.01, P < 0.05). The pain score in either the combined therapy group or Xiaojie point group was lower than that in the tendon-regulation manipulation group after treatment (0.20 -/+ 0.41, 0.15 +/- 0.37 vs 0.60 +/- 0.50, both P < 0.05). Swelling score in the Xiaojie point group was different significantly from that in the tendon-regulation manipulation 0.49 vs 06.4 vs. 20+0.41, P < 0.05). The different value of pain score in either the combined therapy group or Xiaojie point group was higher than that in the tendon-regulation manipulation group before group after treatment (0.65 bined therapy group or Xiaojie point group was higher than that in the tendon-regulation manipulation group before and after treatment (2.35 +/- 0.59, 2.45 +/- 0.51 vs 2.00 +/- 0.46, both P < 0.05). The different value of swelling score in the tendon-regulation manipulation group was higher than that in the Xiaojie point group before and after treatment (2.30 +/- 0.57 vs 1.60 +/- 0.60, P < 0.05).
CONCLUSIONAcupuncture at Xiaojie point combined with tendon-regulation manipulation achieve an apparent effect of analgesia and detumescence on ankle sprain.
Acupuncture Analgesia ; Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Ankle Injuries ; physiopathology ; therapy ; Combined Modality Therapy ; Female ; Humans ; Male ; Tendons ; physiopathology ; Therapy, Soft Tissue ; Young Adult
6.Metatropic dwarfism in a case.
Ji HUANG ; Li-fu GUAN ; Hui-ping SHI ; Wei YU
Chinese Journal of Pediatrics 2003;41(4):310-310
7.Epidemilogical investigation of pneumoconiosis characteristics in Ningxia from 2002 to 2006.
Qi WANG ; Fu-Hai MA ; Guan-Mei WANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2008;26(12):742-743
Adult
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Aged
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Aged, 80 and over
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China
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epidemiology
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Humans
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Middle Aged
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Pneumoconiosis
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epidemiology
8.Use of OTSC in Treatment of Gastrointestinal Bleeding
Fu GUAN ; Zhong CHEN ; Ling ZHONG ; Mingqing ZHANG
Chinese Journal of Gastroenterology 2017;22(3):187-189
Gastrointestinal bleeding is a common complication of gastrointestinal diseases and gastrointestinal surgery, which may lead to hemorrhagic shock or cause death if not treated properly and promptly.Currently, endoscopic treatment for gastrointestinal bleeding includes clip closure, endoscopical injection of saline/adrenaline, argon plasma coagulation (APC), electrocoagulation and heater probe coagulation etc..Endoscopic clip closure mainly includes traditional closure through-the-scope clip (TTSC) and more novel closure over-the-scope clip (OTSC).This article reviewed the use of OTSC in the treatment of gastrointestinal bleeding.
9.Inhibitory effects of TLR7 on Mycobacterium tuberculosis
Ruifang LI ; Zhiyu GUAN ; Yurong FU ; Zhengjun YI
Chinese Journal of Zoonoses 2014;(12):1223-1226
ABSTRACT:The aim of the present study was to investigate the inhibitory effects of TLR7 on Mycobacterium tuberculosis . TLR7 on infected RAW264 .7 cells was activated by chemical synthesis of TLR7 activation motif ssRNA .Activated RAW264 .7 cells were inoculated with Mycobacterium tuberculosis ,quantitative PCR method was applied to detect the phagocytosis rate of cell to bacteria at different time after infection .Cytokine production was measured by ELISA from cell supernatant .Cells were cultured on Roche medium and counted after sterile cracked with TritonX‐100 and diluted with PBS .Scanning electronic micro‐scope ( SEM ) was applied to detect the morphological changes of cells treated with TLR7 activation motif ssRNA .The highest phagocytosis rate of bacteria of RAW264 .7 cells was at 3 hours post infection (P>0 .05) .Compared with that of the control group ,treatment after 36 hours intracellular bacterial quantity in ssRNA treated group was lower (P<0 .05) ,levels of IL‐12 (P<0 .05) and IL‐4 (P<0 .05) were increased .For treatment after 48 hours ,level of IL‐4 (P<0 .05) was decreased ,and TNF‐α (P<0 .05) was increased .For treatment after 3 hours ,cell morphology of the ssRNA group was obviously better than the control group and appeared lots of phagosomes .Results suggested that TLR7 could enhance macrophages in killing Myco‐bacterium tuberculosis by forming phagosomes and regulating cytokines production ,and TLR7 activation motif ssRNA could be used in the treatment of tuberculosis .
10.Effect of continuous blood purification on immune function and prognosis in patients with severe sepsis
Suzhen FU ; Jie SUN ; Yun DONG ; Qi ZHAO ; Bingxing GUAN
The Journal of Practical Medicine 2014;(17):2731-2734
Objective To investigate the effect of continuous blood purification (CBP) on pro-or anti-inflammatory immune function and prognosis in patients with sepsis. Methods One hundred and two patients with sepsis were randomized into the CBP group (60 cases) and the control group (42 cases). The patients in the control group were treated with conventional therapy, and the patients in the CBP group received at least 72 h CBP treatment additionally. The APACHEⅡ score, SOFA score, the 28 day survival rate and ICU length of stay were recorded and levels of spleen Th1, Th2 were assessed by FACS flow cytometry. Enzyme linked immunoadsordent assay (ELISA) was used to determine the levels of serum IL-1, IL-10 and TNF-α before and at 24, 48, 72 h after the treatment. Results The APACHEⅡscore and SOFA score decreased markedly in the CBP group after the treatment (Р <0.05). The period of staying in ICU of patients in the CBP group was shorter than that of patients in the control group (Р < 0.05). There was no significant difference of the 28 day survival rate between the two groups (91.6% vs 71.2%, Р > 0.05). Compared with the control group, levels of IL-1, IL-10 and TNF-α were decreased markedly, and the ratio of Th1 / Th2 was increased significantly at 72 h after the treatment in the CBP group (Р < 0.05). Conclusion CBP can eliminate inflammatory mediators, and help to enhance the immune function, and restore the balance of Th1 / Th2 in patients with severe sepsis.