2.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
3.Experience of transurethral resection of prostate for larger prostatic hyperplasia
Hua HU ; Lian FU ; Chaoying LIU ; Yue WU ; Yi TAN
Chinese Journal of Geriatrics 2012;31(11):947-948
Objective To explore the safety and effectiveness of transurethral resection of the prostate (TURP) for large benign prostatic hyperplasia.Methods A total of 56 cases of patients with large benign prostatic hyperplasia were analyzed retrospectively.Results The weight of prostate tissue resected with TURP was 60.2 g (48-122 g).The resection rate was 60%,operation time 113.4 min (70-180 min).The lower urinary tract symptoms (LUTS) after TURP was improved significantly.The international prostate symptom scores (IPSS) of 38 cases were decreased from (25.6±1.5) before operation to (6.0± 2.0) after operation.The maximum urine flow rate were increased from (5.0±1.9) ml/s to (18.0±4.5) ml/s.The mean residual urine volume of 22 cases were reduced from 95 ml (40-250 ml) to 10 ml (0-60 ml).Conclusions Skilled TURP technology for large benign prostatic hyperplasia is a safe treatment with good effect and low complications rates.
4.Preparation of solid lipid nanoparticles loaded with all-trans retinoic acid and their evaluation in vitro and in vivo.
Lian-Dong HU ; Xing TANG ; Fu-De CUI
Acta Pharmaceutica Sinica 2005;40(1):71-75
AIMTo prepare solid lipid nanoparticles (SLN) loaded with all trans retinoic acid using an ultrasonic technique with Compritol 888 ATO as matrix material, and investigate properties of nanoparticles in vitro and in vivo.
METHODSUltrasonic technique was adopted to prepare solid lipid nanoparticles in an aqueous system using all-trans retinoic acid (ATRA) as a model drug. Physicochemical proterties of SLN were investigated in detail. Drug release from two sorts of ATRA-SLN was investigated using a dialysis bag method. Compared with ATRA solution, the in vivo pharmacokinetics of two sorts of ATRA-SLN after intravenous injection to rats were studied.
RESULTSSolid lipid nanoparticles loaded with all-trans retinoic acid was readily and quickly prepared by ultrasonic technique. The morphological investigation by Transmission Electron Microscopy (TEM) showed that the particles had round and uniform shapes. The mean diameters of them were (158 +/- 9) nm and (89 +/- 11) nm separately. The SLN dispersion was stable at 4 degrees C for more than one year. Drug loading was 3.3%, drug entrapment efficiency was more than 95%, the in vitro release was well in accordance with Weibull distribution. Compared with ATRA control solution, SLN could stay in the blood circulation for a longer time after intravenous injection.
CONCLUSIONThe ultrasonic technique was appropriate for the preparation of solid lipid nanoparticles.
Animals ; Drug Carriers ; Drug Delivery Systems ; Drug Stability ; Fatty Acids ; Male ; Nanostructures ; chemistry ; Particle Size ; Poloxamer ; chemistry ; Polysorbates ; Rats ; Rats, Wistar ; Tretinoin ; administration & dosage ; pharmacokinetics ; Ultrasonics
6.The establishment of diabetic peripheral neuropathy model in spontaneous diabetic GK rats
Xiaoxu FU ; Lulin FENG ; Xiyu ZHANG ; Yun HU ; Xiaoya NAN ; Chunguang XIE ; Lian DU
Tianjin Medical Journal 2016;44(8):989-992
Objective To establish a simple diabetic peripheral neuropathy (DPN) rat model with the high fat-fed in GK rats. Methods A total of 30 GK rats (7-8 weeks) were fed with high-fat diet to establish the DPN model. Thirty normal Wistar rats were fed with ordinary diet (control group). The blood-sugar value, body mass, water-intake and food-intake were monitored every week in two groups. The serum level of glycosylated hemoglobin, the right sciatic nerve conduction velocity were detected at 8, 12 and 16 weeks respectively. The left sciatic nerve was used for HE and TUNEL staining. Results The manifestations of polydipsia, polyphagia and growth retardation were gradually appeared in GK rats. After 12 and 16 weeks, the blood-sugar and glycosylated hemoglobin were significantly increased in GK rats compared with those of normal Wistar rats (P < 0.01). The sensory nerve conduction velocity decreased obviously (P < 0.01). And motor nerve conduction velocity showed a certain decline trend (12 week P < 0.05,16 week P > 0.05). The sciatic nerve pathological features and Schwann cell apoptosis suggested that the model of DPN was successfully established (apoptosis index, P <0.01). Conclusion GK rats fed by high-fat diet are the satisfactory models of the DPN in experimental research. And 12-week is a suitable and economical time for molding.
7.The relationship between social anxiety and self-esteem, self-awareness in grade 4-6 primary students
Zhili ZOU ; Pinhong CHEN ; Huaqing MENG ; Hui WANG ; Qinghua LUO ; Yixiao FU ; Lian DU ; Haitang QIU ; Tian QIU ; Hua HU
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(5):436-439
ObjectiveTo explore the status of social anxiety and study the relationship between social anxiety and self-consciousness,self-esteem among primary students.MethodsUsing stratified randomization sampling method,568 primary students in grade 4 ~ 6 were investigated with social anxiety scale for children( SASC ),self-consciousness scale for children(PHCSS) and self-esteem scale(SES).Results①The score of SASC in the group of primary students was obviously higher than that of Chinese norm (P<0.01).②The SASC and negative evaluation scores in the female were higher than the male (P < 0.05 ),the negative evaluation scores in the sixth grade (3.22 ± 2.82) was higher than the fifth grade (2.53 ± 2.76,P < 0.01 ) and fourth grade ( 2.54 ± 2.68,P< 0.05 ).The primary students in migrant workers elementary school,the single parent family and the relatively low income family had higher social anxiety scores than others(P<0.05 ).③The SASC and two major factors were significantly negative correlated with the self-consciousness and each subscale( r =-0.202 ~ -0.511,P < 0.01 ),and positive correlated with the self-esteem( r =0.282,0.270,0.305,P < 0.01 ).④)The anxiety and gregariousness had significant prediction function of 25.5% on negative evaluation,and 28.2% on SASC,anxiety,gregariousness,intelligence and school situation had significant prediction function of 20.4% on social avoidance and distress.ConclusionsPrimary students in grade 4 ~ 6 existed social anxiety.There are significant differences for their level of anxiety in different gender,grade,elementary school,family structure,monthly income.And social anxiety is closely related to self-consciousness and self-esteem.
8.Relationship of Dopamine ?-hydroxylase Gene with Qualitative and Quantitative Characters of Schizophrenia
Pinhong CHEN ; Qing QIN ; Ying OU ; Yixiao FU ; Hua HU ; Qinghua LUO ; Lian DU ; Haitang QIU ; Huaqing MENG
China Pharmacy 2005;0(18):-
0.05). The factor score of "preoccupation"in patients with Ins/Del genotype was higher than that with Ins/Ins and Del/Del genotypes(P=0.052),as well as in male patients of experimental group(P=0.052). CONCLUSION: The results suggest that D?H gene-4784-4803del polymorphism may not be associated with qualitative and quantitative characters of schizophrenia. Male schizophrenia patients with Ins/Del encoding D?H gene-4784-4803del are significantly characterized with the character of "preoccupation".
9.Therapeutic effects of leaf extract from Cyclocarya paliurus on rats with type 2 diabetes mellitus
Lijun FU ; Minghua HU ; Xiquan YIN ; Dawei LIAN ; Wenkang REN ; Yifei XU ; Pinglong FAN ; Yaqi CHEN ; Yongfeng CAI ; Hongying CAO
Chinese Traditional Patent Medicine 2017;39(6):1134-1138
AIM To speculate the hypoglycemic mechanism for rats with type 2 diabetes by exploring the therapeutic effects of leaf aqueous extract from Cyclocarya paliurus on liver insulin receptor (InsR) and insulin receptor substrate 2 (IRS-2).METHODS The diabetic rat model was established through intraperitoneal injection of streptozotocin and fed with high-fat diet.The moleled rats were equally assigned into the control group and leaf aqueous extract from Cyclocarya paliurus group (extract group).After the test extract was orally administrated for four weeks,body weight,urine output,food intake,water intake and fasting blood-glucose (FBG) were measured,and the levels of serum insulin,InsR and IRS-2 mRNA in liver tissue were investigated in rats.RESULTS Compared with the control group,the extract group showed a reduction in urine output,food intake,water intake,FBG and insulin levels.Meanwhile,the rats' body weights in extract group were presented a trend to increase.The gene expressions of InsR and IRS-2 in liver tissue were up-regulated.Moreover,the insulin sensitivity was improved.CONCLUSION The leaf aqueous extract from Cyclocarya paliurus can reduce FBS,improve insulin sensitivity,which may be associated with the increase of InsR and IRS-2 gene expression in liver tissue.
10.Effects of Nrf2 on gamma-glutamylcysteine synthase in lung of guinea pigs with bronchial asthma.
Yun-Fu ZHUN ; Ai-Guo DAI ; Rui-Cheng HU ; Yong-Lian JIAN
Chinese Journal of Applied Physiology 2006;22(4):492-496
AIMTo investigate the effects of Nrf2 (Nuclear-E2 related factor) on gamma-glutamylcysteine synthase (gamma-GCS) in lung of guinea pigs with bronchial asthma.
METHODS20 adult male guinea pigs were randomly divided into two groups (n = 10): control group (C group) and asthmatic group (A group), asthmatic model was established by ovalbumin intraperitoneal and ovalbumin inhalation. The reactive oxygen piece (ROS), reduced glutathione (GSH), oxidant glutathione (GSSG) and total GSH in lung tissue were examined respectively. Inflammatory cell infiltration and index of remodeling of bronchiole were detected. In situ hybridization detected the gamma-GCS heavy subunit (gamma-GCS h) mRNA in lung tissue. Immunohistochemistry detected the expression of Nrf2 protein and gamma-GCS protein in lung tissue. RT-PCR measured the expression of Nrf2 mRNA in lung tissue. The activity of gamma-GCS was measured by coupled enzyme assay.
RESULTS(1) The number of eosinophils and lymphocytes in bronchiole of A group were significantly higher than that of C group (P < 0.05), the remodeling of bronchiole in A group was definite. (2) ROS (U/mg pro), GSSG (micromol/g pro) and total GSH in lung tissue of A group were significantly higher than that of C group (P < 0.01). The GSH/GSSG in lung tissue of A group was much lower than that of C group (P < 0.01), GSH in lung tissue showed no difference between A group and C group. (3) Immunohistochemistry indicated that Nrf2 protein and gamma-GCS protein were more positively expressed in A group than that in C group (P < 0.01). In situ hybridization discovered that the expression of gamma-GCS-h mRNA in lung tissue of A group was more positive than that of C group. (4) RT-PCR showed that the expression of Nrf2 mRNA was no difference between A group and C group (P > 0.05). (5) The activity of gamma-GCS of A group was (28 +/- 8)U which was significantly higher than that of C group (9 +/- 2)U (P < 0.01). (6) Linear correlation analysis indicated that in lung tissue of guinea pig with asthma there existed strongly positive relationship among ROS, GSSG and the expression of Nrf2, gamma-GCS mRNA, gamma-GCS protein, the activity of gamma-GCS, there existed strongly negative relationship among GSH/GSSG and the expression of Nrf2, gamma-GCS mRNA, gamma-GCS protein, the activity of gamma-GCS.
CONCLUSIONThere existed oxidative stress in lung of guinea pigs with bronchial asthma, which possibly positively regulated gamma-GCS via up regulating transcription factor Nrf2.
Animals ; Asthma ; metabolism ; Glutamate-Cysteine Ligase ; metabolism ; Guinea Pigs ; Lung ; metabolism ; Male ; NF-E2-Related Factor 2 ; metabolism ; Oxidative Stress ; RNA, Messenger ; genetics