1.Research progress of lactoferrin as drug carriers.
Hui-xian TANG ; Zhen-hai ZHANG ; Zhi-ying ZHAO ; Hui-xia LÜ
Acta Pharmaceutica Sinica 2015;50(6):675-681
Lactoferrin (Lf) is one of the food protein belonged to the innate immune system. Apart from its main biological function of binding and transport of iron ions, lactoferrin also has many other functions and properties such as antibacterial, antiviral, antiparasitic, catalytic, anti-cancer, anti-allergic and radioprotecting. Lf is usually used as additives of food and cosmetics. The research of lactoferrin has been increasingly reported, and the application of lactoferrin as a drug carrier has drawn extensive attention over the recent year. In this paper, researches of lactoferrin as drug carriers are classified and summarized in brain targeting, liver tumor targeting, lung tumor targeting and oral delivery systems according to their different characteristics.
Administration, Oral
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Brain
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Drug Carriers
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Humans
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Lactoferrin
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chemistry
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Neoplasms
2.Expression of protease activated receptors-2 in intestinal mucosa of acute necrotizing pancre-atitis in rats
Liang XIA ; Jiang CHEN ; Pi LIU ; Tao SU ; Ying HU ; Nonghua Lü
Chinese Journal of Digestion 2012;32(9):598-601
Objective To explore the correlation between the expression of protease activated receptors-2 (PAR-2) and intestinal mucosal barrier injury of acute necrotizing pancreatitis (ANP) in rats.Methods The ANP rat model was created.The expression of PAR-2 in rat's intestinal mucosa of sham-operated group and ANP group at six,12 and 24 hours after model established was detected by immunohistochemistry (IHC),reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.The difference between groups was analyzed by one way analysis of variance.Results The results of IHC indicated that PAR-2 expression in rat's intestinal mucosa of sham-operated group was weak.The number of PAR-2 expression positive cells and immunostaining intensity increased significantly after ANP model established.The IHC score was 4.88±0.33,5.87±0.32 and 11.17±0.27 at six,12 and 24 hours after model established respectively.Compared with those of shamoperated group (2.86 ± 0.31),the differences were statistically significant (F=747.08,P<0.01).The expression of PAR-2 at mRNA and protein level in intestinal mucosa of sham-operated group was very low.As time extended after ANP model established,both expression increased gradually.The PAR-2 mRNA was 0.56±0.03,0.69±0.03,1.05±0.05,and the protein was 0.28±0.02,0.35±0.03,0.69±0.04 at six,12 and 24 hours after model established respectively.Compared with shamoperated group,the differences were statistically significant at each time point (F=785.69,1177.82,both P<0.01).Conclusions PAR-2 is activated in the inflammatory progress of ANP,and may play an important role in the pathogenesis and development of intestinal mucosa barrier injury in ANP.
3.Clinical observation on treatment of cervicogenic headache with turtle-probing needling at Tianzhu (BL 10).
Chinese Acupuncture & Moxibustion 2006;26(11):796-798
OBJECTIVETo observe therapeutic effect of turtle-probing needling at Tianzhu (BL 10) on cervicogenic headache.
METHODSSeventy cases were randomly divided into a treatment group (n=36) and a control group (n= 34). The treatment group were treated with turtle-probing needling at Tianzhu (BL 10) on the affected side, first perpendicularly needling the point and then penetration-needling homolateral Fengchi (GB 20), Fengfu (GV 16), below cervical Jiaji points (EX-B2), and occipital tuberosity in order. And the control group were treated with acupuncture at Fengchi (GB 20),Tianzhu (BL 10), Shuaigu (GB 8), Touwei (ST 8) and corresponding cervical Jiaji points (EX-B2), Houxi (SI 3), Ashi points at the affected side with routine needling manipulation. Treatment was given once each day, 6 sessions constituting one course.
RESULTSThere was no significant difference between the two groups in therapeutic effect (P > 0.05), but the cured and markedly effective rates for analgesic effect in the observation group at the first three sessions were higher than those in the control group (P < 0.05).
CONCLUSIONThe turtle-probing needling method and the routine needling have a similar therapeutic effect, but the turtle-probing needling method more rapidly produces the analgesic effect.
Acupuncture Points ; Acupuncture Therapy ; methods ; Adolescent ; Adult ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Post-Traumatic Headache ; therapy
4.Hhigh-frequency ultrasound research on the normal adult sciatic nerve.
Su-Hong SHEN ; Hai-Xia LÜ ; E ZHAN-SEN ; Zhan-Mei LIU ; Ying ZHANG
China Journal of Orthopaedics and Traumatology 2013;26(2):107-110
OBJECTIVETo explore the application value of the high-frequency ultrasound examination of the normal adult sciatic nerve.
METHODSThe high frequency ultrasound was used to examine 60 cases ,a total of 120 normal adult sciatic nerve (28 males and 32 females ranging in age from 19 to 38 years old, averaged 23.8 years old). The age between men and women, height and weight showed no significant difference. The cross-sectional scanning and longitudinal scanning, if necessary, Panoramic Imaging were conducted. The cross section area of sciatic nerve was measured at the hip line, the middle of the thigh, the edge of the popliteal fossa, and the relationship betwen the sciatic nerve and surrounding muscle group was observed in each cross-section. Probe was rotated 90 degree to display the long axis of sciatic nerve, to get the longitudinal section along the long axis from top to bottom. So the sciatic nerve in the vertical axis and the surrouunding muscles was displayed by above method combined with panoramic imaging. The data of coss-sectional area of sciatic nerve were obtained at the central of hip line, the middle of the thigh, the edge of the popliteal fossa, and then were divided into different groups by gender and sides to do statistical analysis.
RESULTSThe applicatic nerve in thigh can be clearly displayed by high-frequency ultrasound in this group of 60 people with 120 nerves, but low display rate of sciatic nerve in hip, approximatley 31.4% (21/66). The longitudinal section of the sciatic nerve displayed as the banded isoechoic or slightly hyperechoic, parallel partial hyperechoic line could be seen in it; in the cross-section, it showed a clear edge of the sieve-like isoechoic or slightly hyperechnie, which could be oval ,trapezoidal, triangular and irregular in shape; the sciatic nerve bad visible passive movement and shape change when muscle contracted. There were statistically significant differences of cross-sectional area of sciatic nerves at the hip line ,the middle of thigh and edge of the popliteal fossa between 56 sciatic nerves in 28 males and 64 sciatic nerves in 32 females (P< 0.05); but there were no significant differences between left and right sides (P>0.05).
CONCLUSIONHigh frequency ultrasound can be used as the first choice of the sciatic nerve when an imaging method preferred.
Adult ; Female ; Humans ; Male ; Sciatic Nerve ; diagnostic imaging ; Ultrasonography
5.Animal trials for mPEG-modified red blood cells.
Yan QIU ; Ying-Xia TAN ; Li-Li LI ; Su-Bo LI ; Qiu-Shuang LÜ ; Yi ZHA ; Yang-Pei ZHANG
Journal of Experimental Hematology 2006;14(4):816-821
This study was aimed to investigate the survival rate and difference of transfused modified and unmodified RBC at 24 hours. The modified and unmodified RBC from mice, monkey, pig and human were labeled by using FITC, then these blood RBCs were transfused to homogeneous and heterogeneous animals. The result showed that 24 hour survival rate of unmodified mice RBC transfused to mice was 74%, while survival rate of 2.0 mmol/L mPEG-SPA modified mice RBC transfused to mice was 45%, difference between them was significant. The 24 hour survived rate of unmodified human RBC transfused to mice was 8%, while 24 hours survival rate of 2.0 mmol/L mPEG-SPA modified human RBC transfused to mice was 5% without statistical difference. The 24 hour survived rate of homogeneous transfusion of modified monkey RBC was 90%, while survival rate of modified human and pig RBC was zero on 24 hours after transfusion to monkey. It is concluded that RBC labeling methods and mice species are unrelated to 24 hours survival rate, but mPEG variety and concentration are related to mouse RBC life-span. It is incredible to use mouse RBC homogeneous transfusion result instead of human RBC to evaluate longevity and safety of modified human RBC. But modified human RBC transfused to mice can be a model to evaluate longevity of modified human RBC. It is very difficult to get the result about modified RBC life span by RBC transfusion among great heterogeneous mammal animals. So evaluation in large mammal animal models needs to be further studied.
Animals
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Cell Survival
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drug effects
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Erythrocyte Transfusion
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methods
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Humans
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Macaca mulatta
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Male
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Mice
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Polyethylene Glycols
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pharmacology
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Swine
6.Regularity of sugar-uptake in human red blood cells.
Guo-Bo QUAN ; Cui-Cui LÜ ; Min-Xia LIU ; Wen-Bo HU ; Yan WANG ; Ying HAN
Journal of Experimental Hematology 2006;14(3):592-596
Lyophilization of human red blood cells has important significance in clinical application. Some sugars, especially trehalose, can be more tolerant of some organism or cells to dry environments, But, how to bring sugars into cells is a challenge. This study was aimed to investigate the regularity of sugar-uptake in human red blood cells. The absorption rate of trehalose and glucose in red blood cells, free hemoglobin level and erythrocyte deformation index were determined at different incubation temperature (4, 25 and 37 degrees C), different sugar concentration (0, 0.2, 0.4, 0.6, 0.8 and 1 mol/L) and different incubation time (1, 3, 5, 7 and 9 hours). The results showed that with increase of temperature and extracellular sugar concentration, the uptake of sugar in red blood cells also increased, the intracellular trehalose and glucose concentrations were over 30 mmol/L and 40 mmol/L respectively. The effects of incubation time on uptake of trehalose and glucose were different. With prolonging of incubation time, the uptake of trehalose showed firstly increase and then decrease, however, the uptake of glucose showed a constant increase. But the loading process had side-effect on free hemoglobin and maximum deformation index (MAXDI) of red blood cells, especially for trehalose, which mainly come from high osmotic pressure. It is concluded that the uptake of sugars in red blood cells is closely dependent on incubation temperature, extracellular sugar concentration and incubation time. In certain condition, the efficiency of sugar uptake is very high, but this process also damages red blood cells so as to affect the application of sugars in lyophilization of red blood cells. The research in the future should focus on how to deal with the relation between cell injury and uptake efficiency of sugar in red blood cells.
Blood Preservation
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adverse effects
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Cryoprotective Agents
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pharmacokinetics
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Erythrocyte Membrane
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drug effects
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Erythrocytes
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metabolism
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Freeze Drying
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Glucose
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pharmacokinetics
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Humans
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Trehalose
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pharmacokinetics
7.The role of aptamer-siRNA nucleic acid compound in promoting the apoptosis of K562 cells
hua Jian LU ; qiong Zhao CUI ; xia Zhao ZHAO ; Ying LÜ ; Yi XU
Journal of Xi'an Jiaotong University(Medical Sciences) 2017;38(6):862-865,871
Objective To observe the effect of aptamer-siRNA nucleic acid compound on the apoptosis of K562 cells in human chronic myelogenous leukemia (CML)and explore its acting mechanisms.Methods K562 cells were transfected with different concentrations of aptamer-siRNA solution.The effects of aptamer-siRNA on the proliferation and apoptosis of K562 cells were detected by MTT method and AnnexinV/PI double staining method,respectively.The effects of aptamer-siRNA on the expressions of bcl-2,Bax and casepase-3 at protein and mRNA levels in K562 cells were detected by Western blot and RT-PCR method,respectively.Results Compared with the control group,the proliferation of K562 cells was significantly inhibited,early apoptosis rate of K562 cells increased significantly,the expression levels of bcl-2 protein and mRNA were significantly decreased,while the expression levels of Bax and caspase-3 protein and mRNA were significantly increased after transfection with aptamer-siRNA (P <0.05).Aptamer-siRNA nucleic acid complex at the concentration of 50 -250 μmol/L)had a significant dose-effect relationship on bcl-2,Bax and caspase-3 mRNA.Conclusion Aptamer-siRNA nucleic acid compound can promote the decreased number of bcl-2 gene and the growth of Bax and caspase-3 genes,thus promoting the apoptosis of K562 cells.
8.Sequence variability of human cytomegalovirus UL144 open reading frame in low-passage clinical isolates.
Rong HE ; Qiang RUAN ; Chang XIA ; Lan-qing LIU ; Sheng-min LÜ ; Ying LU ; Ying QI ; Yan-ping MA ; Qing LIU ; Yao-hua JI
Chinese Medical Sciences Journal 2004;19(4):293-297
OBJECTIVETo explore the relationship between human cytomegalovirus (HCMV) UL144 sequence variability and clinical disease.
METHODSHCMV UL144 open reading frame (ORF) was amplified by PCR assay in 72 low-passage isolates [65 congenitally infective children and 7 healthy children who were HCMV-DNA positive by quantitative PCR (qPCR)]. All positive PCR products were analyzed by heteroduplex mobility assay and single-stranded conformation polymorphism (HMA-SSCP) and 32 of them were sequenced.
RESULTSFifty-five patient isolates and five healthy children isolates were HCMV-UL144 positive by PCR. Sequencing and HMA-SSCP analysis showed that significant strain-specific variability was present in the UL144 ORF. Phylogenetic analysis indicated that the nucleotide sequences could be separated into 3 major genotypes. Comparing between UL144 sequences and the corresponding symptoms showed that genotype 2 did not exist in megacolon isolates. And genotype 1 and 3 were the major types among microcephaly and jaundice isolates respectively.
CONCLUSIONSHCMV-UL144 existed in most of low passage isolates and sequences were hypervariable. The UL144 ORF and its predicted product with the high level of sequence variability in different kinds of isolates suggest that UL144 ORF might play a role in HCMV infectivity and subsequent diseases.
Amino Acid Sequence ; Base Sequence ; Cytomegalovirus ; genetics ; isolation & purification ; Cytomegalovirus Infections ; genetics ; virology ; DNA, Viral ; genetics ; Genetic Variation ; Genotype ; Humans ; Infant ; Infant, Newborn ; Jaundice ; genetics ; virology ; Megacolon ; genetics ; virology ; Microcephaly ; genetics ; virology ; Molecular Sequence Data ; Open Reading Frames ; genetics ; Phylogeny ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Sequence Homology, Amino Acid
9.Recombinant adenovirus vectors carrying antisense MMP2 inhibit invasion of HCC cells in vitro.
Ming-Man ZHANG ; Xing-Hua GOU ; De-Hua LI ; Lü-Nan YAN ; Dong XIA ; Lei HAN ; Zhi SU ; Lai-Ying ZHAO ; Hai-Yang HU
Chinese Journal of Hepatology 2005;13(4):267-270
OBJECTIVESTo construct a recombinant adenoviral vector carrying antisense matrix metalloproteinase-2 (MMP2) and to study its inhibitory effects on the invasiveness and migratory capacity of hepatocellular carcinoma (HCC) cell line HepG2 in vitro.
METHODSTotal RNA was extracted from HCC. Then a 500 bp fragment at the 5' end of the human MMP2 cDNA sequence was synthesized by polymerase chain reaction (PCR) and was reversely inserted into the multiclone site (MCS) of the shuttle plasmid pAdTrack-CMV. With the resultant plasmid and the backbone plasmid pAdEasy-1, the homologous recombination took place in the E.coli BJ5183 and the recombinant adenoviral plasmid carrying the antisense MMP2 gene was constructed. The adenovirus (Ad-MMP2AS) was packaged and amplified in the HEK 293 cells and the viral titer was checked by GFP. Using the Boyden chamber model, the influence of Ad-MMP2AS on the invasion ability of HepG2 cells was determined in vitro.
RESULTSThe recombinant adenovirus vector carrying antisense MMP2 was constructed successfully and a strong green fluorescence was observed in HepG2 cells under a fluorescence microscope. The viral titer was 1 x 10(8); Ad-MMP2AS can effectively inhibit the penetrating capacity of HepG2 cells through Matrigel in vitro.
CONCLUSIONThe recombinant adenovirus with antisense MMP2 can effectively inhibit the invasiveness and migratory capacity of HepG2 in vitro and may have potential in treating HCC.
Adenoviridae ; genetics ; Carcinoma, Hepatocellular ; pathology ; Genetic Vectors ; Humans ; Liver Neoplasms ; pathology ; Matrix Metalloproteinase 2 ; biosynthesis ; genetics ; pharmacology ; Neoplasm Invasiveness ; Oligonucleotides, Antisense ; biosynthesis ; genetics ; pharmacology ; RNA, Antisense ; genetics ; pharmacology ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Tumor Cells, Cultured
10.Molecular mechanism of indirubin-3'-monoxime and Matrine in the reversal of paclitaxel resistance in NCI-H520/TAX25 cell line.
Su-xia LUO ; Wen-ying DENG ; Xin-feng WANG ; Hui-fang LÜ ; Li-li HAN ; Bei-bei CHEN ; Xiao-bing CHEN ; Ning LI
Chinese Medical Journal 2013;126(5):925-929
BACKGROUNDMultidrug resistance (MDR) is a main reason for paclitaxel (TAX) treatment failure. Indirubin-3'-monoxime (IRO) and Matrine are traditional Chinese medicines, which may reverse the resistance of tumor cells to some chemotherapy drugs, but the relationship between paclitaxel resistance and Matrine is still unclear. The aim of this study was to explore the potential molecular mechanism of IRO and Matrine in reversal of TAX resistance.
METHODSIn this study, MTT assay was used to measure the non-cytotoxic dosage of IRO and Matrine on NCI-H520/TAX25 cells and determine the reversal extent of TAX resistance under non-toxic doses. In addition, RT-PCR and Western blotting were used to evaluate the mRNA expression and the protein level of survivin, Oct-4, and Sox-2 in NCI-H520/TAX25 cells using semi-quantitative methods.
RESULTSThere was no obvious inhibition on sensitive cell strains and drug-resistant strains, when the final concentration was at lest 4 µmol/L for IRO and 100 µmol/L for Matrine. So 4 µmol/L of IRO and 100 µmol/L of Matrine were considered as the reversal dosage. When 4 µmol/L of IRO or 100 µmol/L of Matrine were used together with TAX, the sensitivity to TAX increased evidently in NCI-H520/TAX2 cells; the reversal rate of IRO and Matrine was about 1.92 (43.56/22.6 nmol/L) and 1.74 (43.56/25.0 nmol/L), respectively. The mRNA expression and the protein level of survivin, Oct-4, and Sox-2 in NCI-H520/TAX25 decreased significantly (P < 0.05) after addition of IRO or Matrine in TAX treatment, compared to that of TAX treatment alone.
CONCLUSIONThe decrease in both mRNA expression and protein level of survivin, Oct-4, and Sox-2 might be the molecular mechanism, by which IRO and Matrine mediate the reversal of TAX resistance.
Alkaloids ; pharmacology ; Blotting, Western ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Humans ; Indoles ; pharmacology ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Octamer Transcription Factor-3 ; genetics ; metabolism ; Oximes ; pharmacology ; Paclitaxel ; pharmacology ; Quinolizines ; pharmacology ; SOXB1 Transcription Factors ; genetics ; metabolism