AIMTo study the transfection and anti-hepatitis B virus (HBV) effect of the co-modified hepatocytes-targeting cationic liposomes encapsulating anti-HBV antisense oligonucleotides (asON) , and to investigate the transfection mechanisms of the liposomes.

METHODSDipalmitoylphosphatidylcholine (DPPC) and 3beta-[N-(N',N'-dimethylaminoethane)-carbamoyl] cholesterol (DC-Chol) were used as the lipids, beta-sitosterol-beta-D-glucoside (sito-G) and Brij 35 were used to modify the liposomes. Flow cytometry (FCM), fluorescence microscopy and enzyme-linked immunosorbent assay (ELISA) were utilized to evaluate the transfection improvement of the asON encapsulated in the liposomes in primary rat hepatocytes and the antigens inhibition activity in HepG 2.2.15 cells. The transfection mechanisms were evaluated based on the influence of wortmannin, nigericin, and asialofetuin on the antigens inhibition in HepG 2.2.15 cells by ELISA.

RESULTSThe co-modification with sito-G and Brij 35 significantly improved the transfection of the liposomes in primary rat hepatocytes and antigens inhibition effect in HepG 2.2.15 cells. Both transfection efficiency and antigens inhibition effect showed to be concentration-dependent with the asON-encapsulating liposomes. In fluorescence microscopy, the transfected cells showed strong fluorescence in primary rat hepatocytes, especially in the nuclei. Wortmannin, nigericin and asialofetuin decreased the antigens inhibition of the asON-encapsulating liposomes to different levels. Cationic liposomes modification with sito-G and Brij 35 could improve the transfection and antigens inhibition effect of the asON. The transfection mechanisms of the co-modified liposomes included endocytosis and membrane fusion. The ligand sito-G was confirmed to be able to enhance asialoglycoprotein receptor (ASGPR)-mediated endocytosis.

CONCLUSIONCo-modified hepatocytes-targeting cationic liposomes would be a specific and effective carrier to transfer asON into hepatocytes.

Androstadienes ; pharmacology ; Animals ; Asialoglycoproteins ; pharmacology ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Cell Survival ; Cells, Cultured ; Endocytosis ; drug effects ; Female ; Fetuins ; Flow Cytometry ; Hepatitis B Antigens ; metabolism ; Hepatitis B virus ; genetics ; immunology ; Hepatocytes ; cytology ; drug effects ; metabolism ; Humans ; Liposomes ; Microscopy, Fluorescence ; Nigericin ; pharmacology ; Oligonucleotides, Antisense ; chemistry ; genetics ; Polyethylene Glycols ; chemistry ; Rats ; Rats, Wistar ; Sitosterols ; chemistry ; Transfection ; methods ; alpha-Fetoproteins ; pharmacology

Objective To evaluate the knowledge,attitude and practice(KAP)related with AIDS of local floating population.Methods A total of 1 942 floating persons were sampled by multistage cluster sampling and interviewed with questionnaire in the agricultural markets,construction sites,and their habitats within 3 communities in Hongkou District.Results 82.9% of the interviewees were 20 ～49 years old.The average score was 34.77 ±3.52 (maximum score is 44)for knowledge,12.11 ±2.32(maximum score is 19)for attitude,5.50 ±0.95(maximum score is 7)for practice,respectively.The scores increased with educational level significantly.The scores of knowledge were 33.26 ±3.54,34.63 ±3.23,36.56 ±3.39 among the subjects with educational levels of primary school and below,junior high school,senior high school and above.The scores of attitude were 13.77 ±2.27,14.79 ±2.39,15.62 ±2.38,respectively.And the score of practice was 5.40 ±0.90,5.51 ±0.93,5.58 ±1.03,respectively.Conclusion In the present,the KAP relating AIDS of local floating population is poor in Hongkou District.A variety of intervention methods and operational patterns should be developed for the floating population with different educational levels and jobs.

AIMTo prepare fluorescein sodium (FS) cationic liposomes and investigate the influence of cationic lipid (DC-chol) and polyethylene glycol (PEG) with different molecule weight (MW) on cationic liposome incorporation efficiency, cellular delivery and fluidity of liposome membrane.

METHODSUsing FS as a model material for encapsulation, the liposomes were prepared and separated (by sephadex G-50 1 cm x 20 cm column), and the liposome incorporation efficiencies was measured. The interaction between the FS and cationic liposomes was investigated by measuring the change of fluorescent spectrum. The cellular uptake of different liposome forms by choosing HepG2 2.2.15 as an in vitro cell culture assay model, and the influence of PEG on the fluidity of liposome membrane with the technique of fluorescence polarization were investigated.

RESULTSCationic lipid and different PEGs showed great effects on increasing liposome incorporation efficiency (from 0.64% to 86.57%), cellular uptake (from 2.18% to 48.46%) and fluidity of liposome membrane. The effect of PEG was MW dependent, and with the increase of MW, the incorporation efficiency and transfection was improved, and the fluidity of liposome membrane increased.

CONCLUSIONAddition of cationic lipid and high MW PEG into cationic liposomes can enhance the cellular delivery and fluidity of cationic liposomes. Also, they can improve the incorporation efficiency of cationic liposomes.

Cholesterol ; analogs & derivatives ; pharmacology ; Drug Delivery Systems ; Fluorescein ; Hepatoblastoma ; pathology ; Humans ; Liposomes ; Liver Neoplasms ; pathology ; Membrane Fluidity ; drug effects ; Molecular Weight ; Polyethylene Glycols ; chemistry ; pharmacology ; Transfection ; methods ; Tumor Cells, Cultured

OBJECTIVETo develop a method for the determination of Chonglou saponin I and Chonglou saponin II in Yifu ointment.

METHODThe chromatographic separation was performed on Hypersil ODS2 C18 column (4.6 mm x 150 mm, 5 microm) and IBBM-612111 ODS C18 guard column. Acetonitrile-0.02% phosphoric (43:57) as mobile phase. The flow rate was 1 mL min(-1) and column temperature was set at 40 degrees C. The UV detection wavelength was set 203 nm.

RESULTChonglou saponin I showed a good linear relationship at a range of 0.1024-3.2 microg, r =0.9998, the average recovery was 97.4%, and RSD was 1.8% (n = 6); Chonglou saponin II showed a good linear relationship at a range of 0.064-2.0 microg, r = 0.9999, the average recovery was 101.4%, and RSD was 1.0% (n =6).

CONCLUSIONThe method is accurate with the good reproducibility and can be used for the quality control of Yifu Ointment.

Chromatography, High Pressure Liquid ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Liliaceae ; chemistry ; Ointments ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Rosaceae ; chemistry ; Saponins ; analysis

AIMTo construct a liposomal liver targeting delivery system by adding soybean-derived sterylglucoside (SG) to the cationic liposomes.

METHODSThe physico-chemical properties of SG modified cationic lipsomes were investigated using fluorescein sodium (FS) as a model drug, as well as the interaction of SG modified liposomes with HepG2 2. 2. 15 cells in the point of involvement of asialoglycoprotein receptor (ASGP-R) mediated transfection. Liver targeting of modified cationic liposomes were also investigated using liver perfusing technique, and hepatocytes and non-hepatocytes were separated and examined after perfusing.

RESULTSAll the formula yielded high incorporation efficiency (83.12% - 91.74%), small particle size (93.0 - 124.4 nm). The zeta potential of blank liposomes all showed positive values. The transfection efficiency of FS entrapped in SG-liposomes with HepG2 2.2. 15 was significantly higher than that of liposomes without modification. The transfection of SG-liposomes were reduced significantly by the 20/30 mmol galactose as a competitor of ASGP-R. All the cationic liposomes showed high level of liver uptake of FS. Compared with the uptake of non-hepatocytes of each respectively, only SG/Brij-35 liposomes showed difference in FS uptake by hepatocytes (P < 0.05).

CONCLUSIONIt showed that SG/Brij-35 modified cationic liposomes are potentially useful drug carrier to liver but may be affected by different modification.

Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cations ; pharmacokinetics ; Cell Line, Tumor ; Cholestenes ; administration & dosage ; pharmacokinetics ; Drug Delivery Systems ; Galactose ; pharmacology ; Humans ; Liposomes ; Liver ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Male ; Particle Size ; Polyethylene Glycols ; administration & dosage ; pharmacokinetics ; Rats ; Transfection

Objective To observe the therapeutic efficacy of hyperbaric oxygen (HBO) for focal cerebral infraction influenced by the treatment time after permanent middle cerebral artery occlusion (MCAO) in rabbits. Methods Seventy-five male rabbits were randomly divided into simple MCAO group (n=25), MCAO+HBO group (100% O2, 250 kPa, 1 h/d, from 1 d after MCAO, n=25) and MCAO+DHBO group (100% O2, 250 kPa, 1 h/d, from 7 d after MCAO, n=25). Behaviors and volumes of infarction were observed, and microdialysis was applied to monitor the concentrations of glucose, lactate, pymvate and glutamate around the infarct foci at 1, 3, 10 and 30 d after permanent MCAO. Results Behaviors'score was lower in MCAO+HBO group than the others (P<0.05). The infarct volume from day 3 to day 30 was significantly smaller in MCAO+HBO group than in the other 2 groups (P<0.05). The lactate and pyruvate ratio was increased after MCAO in three groups, but they were lower in the MCAO+HBO group than in the others at day 1 and day 3 (P<0.05). The glutamate concentration was increased after MCAO, peaked at 3 d, but at day 1 and day 3 the glutamate concentration was lower in the MCAO+HBO group (P<0.05). Conclusions HBO treatment could protect the brain from infarction through improving the energy metabolism and decreasing the excitatory amino acids disorders around the infarct foci after MCAO in rabbits. In order to improve the therapeutic efficacy of HBO, it should be performed as possible.

OBJECTIVETo establish the quality control standard of Xindi soft capsule.

METHODQuercetin, kaempferol and isorhamnetin were isolated by TLC with chloroform-ethyl formate-formic acid (5:4:1). The chromatographic separation was performed on a Diamonsil C18 column (4.6 mm x 250 mm, 5 microm). Acetonitrile-water-phosphoric (30:70:0.1) as mobile phase. The flow rate was 1 mL x min(-1) and column temperature was set at 40 degrees C. The UV detection wavelength was set at 254 nm.

RESULTQuercetin, Kaempferol and Isorhamnetin could be identified by TLC. Quercetin showed a good linear relationship at a range of 0.412-1.648 microg, r = 0.999 9, the average recovery was 96.8%, and RSD was 0.9% (n = 6). Kaempferol showed a good linear relationship at a range of 0.021-0.083 microg, r = 0.999 8, the average recovery was 96.9%, and RSD was 2.0% (n = 6). Isorhamnetin showed a good linear relationship at a range of 0.183-0.732 microg, r = 0.999 9, the average recovery was 97.1%, and RSD was 1.6% (n = 6).

CONCLUSIONThe method is accurate with the good reproducibility and can be used for the quality control of Xindi soft capsule.

Capsules ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; standards ; Flavonols ; analysis ; Hippophae ; chemistry ; Kaempferols ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Quercetin ; analysis ; Reproducibility of Results

AIM To investigate the dose-proportion relation of Tibetan medicine Siwei Jianghuang Prescription (SJP) for protective effects on diabetic nephropathy (DN),and the underlying mechanism.METHODS Diabetes mellitus rats induced by intraperitoneal injection of streptozotocin (STZ) (60 mg/kg) were randomly divided into model group,metformin support group,and eight SJP groups with dose-proportion variation (with reference to the uniform design method) for corresponding drug administration once a day,for four weeks.Measurement of fasting blood glucose (FBG) by a blood glucose meter,the concentrations of blood urea nitrogen (BUN),uric acid (UA),serum creatinine (SCr) and total protein (TP) by chemical methods,serum transforming growth factor-β1 (TGF-β1) and vascular endothelial growth factor (VEGF) by ELISA kits were conducted,the pathological morphology observation and glomerular basement membrane thickness detection by electron microscope were accomplished as well.Principal components analysis (PCA) and multivariate progressive regression analysis (MSRA) were employed to analyze the relationship between the dose-proportion to pharmacodynamics.RESULTS The resultant indexes revealed variant pharmacological improvement in each treatment group.MSRA results showed that the levels of BUN,renal index,FBG,glomerular basement membrane thickness,VEGF,Scr,and UA had correlative relations with a multiple linear or a multiple non-linear in all groups,which regression equation had a statistical significance (P ＜ 0.05);TGF-β1 level and total protein index with the dose-proportion had no linear or non-linear relation,which the regression equation statistical showed non-significance (P ＞ 0.05).In the global optimization comparison around the range of uniform design,the optimal dosage of the rats model was Curcumae Longae Rhizoma ∶ Berberidis dictyophyllae Cortex ∶ Phyllanthi Fructus ∶ Tribuli Fructus =1 ∶ 2 ∶ 1 ∶ 2.CONCLUSION Siwei Jianghuang Prescription shows better therapeutic effects on DN,which may be related to reducing the levels of BUN,renal index,FBG,glomerular basement membrane thickness,VEGF,Scr and UA.

Objective To investigate the changes of matrix Metalloproteinases-9 (MMP-9) content in the cerebrospinal fluid of patients with brain injury and its clinical significance.Methods Sixty cerebrospinal fluid (CSF) specimens from patients with traumatic brain injury were chosen in our study as experimental group,and other CSF samples from 35 healthy subjects were selected as control group.The content of MMP-9 in cerebrospinal fluid was measured by enzyme-linked immunosorbent assay (ELISA) at 24 h and 72 h,1 and 2 weeks,respectively; and the comparative study was performed.Results The MMP-9 content in all CSF samples from the experimental group was significantly higher than that in the healthy control group (P＜0.05).Conclusion MMP-9 content in CSF of patients with brain injury increases obviously; MMP-9 in CSF may contribute to the severity of traumatic brain injury determination and guide the treatment strategies,which is worth for further study.

The establishing and improving of quality standards for ethnic medicine have close relation with the safety and efficacy of clinical medicine. It is significant to promote the modernization and industrialization of ethnic medicine. This paper summarized the present status of quality standards for ethnic medicine, and pointed out the deficiency of ethnic medicine; the origin of medicinal materials was not clear and varieties were confused; the safety research of mineral drug was weak and so on. Based on these, firstly, the research of quality standards for ethnic medicine should be under the guidance of ethnic traditional medical theory and clinical experience and strengthen the quality control and evaluation of safety and effectiveness for ethnic medicine. Further, much attention should be payed to protection of natural medicinal resources and sustainable utilization. Lastly, modern chemical analysis technology shouled be used during the research process on the basis of ethnic characteristics. It will promote the establishment and improvement of quality standards for ethnic medicine and will be benificial to modernization and industrialization development of ethnic medicine.
Drugs, Chinese Herbal ; chemistry ; standards ; Humans ; Medicine, Chinese Traditional ; standards ; Plants, Medicinal ; chemistry ; growth & development ; Quality Control