1.Preparation of doxorubicin-loaded nanoparticles based on modified popaccharide and its targeting effect on hepatocellular carcinoma cells
Yang BAI ; Qingqing XIONG ; Hai WANG ; Liyun PANG ; Tianqiang SONG
International Journal of Biomedical Engineering 2017;40(1):-
Objective To prepare a redox-responsive doxorubicin-loaded nanoparticle,and to study its in vitro realease behavior and targeting effect on heptoma cells.Methods Cystamine was grafted on the side chains of hyaluronic acid with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide catalyst,and then β-cyclodextrin (β-CD) was conjugated on the amine groups of the cystamine by Schiff's base reaction to prepare β-CD modified hyaluronic acid (HACD).The HACD/DOX nanoparticles were prepared by encapsulating DOX into HACD using dialysis method.The drug loading,encapsulation efficiency,particle size and distribution,zeta potential and other physical and chemical properties,as well as in vitro drug release behavior of the HACD/DOX nanoparticles were characterized.The cytotoxicity of HACD/DOX nanoparticles to HepG2 cells was studied by cell counting kit-8 (CCK-8) method.The targeting effect of HACD/DOX nanoparticles on HepG2 cells was studied using flow cytometry and confocal laser scanning microscopy (CLSM).Results HACD were successfully synthesized,which could carry DOX to form uniform homogeneous nanoparticles.The drug loading of DOX in the nanoparticles was (16.1±0.2)% and the encapsulation efficiency was (64.2±0.9)%.The transmission electron microscope images indicated that the shape of the HACD/DOX nanoparticles was homogeneous sphere.The results of granularity analysis showed that the average size of the HACD/DOX nanoparticles was (203.1 ±2.5) nm with a narrow size distribution (PDI =0.202).The zeta potential of the HACD/DOX nanoparticles was (-29.1±0.8) mV.The in vitro release behavior of the nanoparticles exhibited obvious redox-sensitivity.The results of in vitro cytotoxicity showed that the blank carrier material HACD had no obvious toxicity to hepatoma cells,and the HACD/DOX nanoparticles could effectively kill hepatoma cells with the 0.38 μg/ml half maximal inhibitory concentration (IC50) value at 48 h.Flow cytometry and CLSM results demonstrated that the HACD/DOX nanoparticles could target hepatoma cells through the mediating effect of hyaluronic acid.Conclusions The prepared HACD/DOX nanoparticles have suitable particle size,high drug loading and encapsulation efficiency,and can release DOX under the stimulation of reducing agent.These nanoparticles have obvious targeting effect on hepatoma cells,which is expected to be applied as the drug delivery system of hepatocellular carcinoma (HCC) therapy.
2.Analysis on envelope gene of type Ⅰ dengue virus isolated from Guangzhou area in 2009
Zhijun BAI ; Yulin WANG ; Biao DI ; Lei LUO ; Yu CHEN ; Liyun JIANG ; Ming WANG ; Zhicong YANG
Chinese Journal of Infectious Diseases 2010;28(11):641-644
Objective To sequence and analyze the envelope (E) gene of type Ⅰ dengue virus isolated from Guangzhou in 2009 for tracing the infection source. Methods The serum samples were collected from patients diagnosed with dengue fever in Guangzhou area during 2009. Dengue virus was isolated and cultured in C6/36 cells.The whole length of E gene was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) and then sequenced. The phylogenetic tree was drawn by neighbor-joining method. The bioinformatics analysis was performed by combining the phylogenetic information and the epidemiology data. Results Four strains of type Ⅰ dengue virus were isolated from 19 samples. E gene of these strains was amplified and sequenced. The phylogenetic analysis showed that 09/GZ/9104 strain and 09/GZ/9236 strain had identical nucleotide sequence and fell within the American/African group, 09/GZ/11534 stain and 09/GZ/11562 strain had similar sequence homology and fell within the Asian group. Conclusion The typeⅠdengue viruses in Guangzhou area in 2009 are imported, which belong to two genotypes and may come from two independent origins respectively.
3.Effects of vibrating the abdomen on peripheral blood gastrointestinal hormones and gastrointestinal motility of young anorexia model rats
Ling ZHOU ; Yan ZHANG ; Nan GU ; Weijie BAI ; Liyun YANG ; Jinlong LI ; Yanping YANG
Journal of Acupuncture and Tuina Science 2024;22(2):104-110
Objective:To observe the therapeutic effect of vibrating the abdomen on anorexia model rats,as well as its effects on cholecystokinin octapeptide(CCK-8)and motilin(MTL)in the peripheral blood. Methods:Forty young rats were randomly divided into a normal group(n=10)and a modeling group(n=30).Rats in the normal group were fed common feed.The anorexia model was established by the etiological simulation method in the modeling group,and these rats were further randomly divided into a drug group,a vibrating abdomen group,and a model group 3 weeks after the anorexia model was induced,with 10 rats in each group.The drug group was given Jian Wei Xiao Shi Pian by intragastric administration at a dose of 0.72 g/(kg·bw)(0.72 g drug was dissolved in 10 mL purified water).The normal group and the model group were given purified water once a day in the morning.The vibrating abdomen group was treated with vibrating the abdomen once a day for 21 times.The body mass,food intake,serum CCK-8,MTL,gastrin(GAS),neurotensin(NT)levels,and the intestinal propulsion rate of rats in each group were measured. Results:Compared with the model group,the body mass,food intake,serum MTL and GAS levels,and the small intestine propulsion rate increased significantly,and the serum CCK-8 and NT levels,the gastric residual rate decreased significantly in the vibrating abdomen group and the drug group(P<0.05).There were no significant differences between the vibrating abdomen group and the drug group(P>0.05). Conclusion:Vibrating the abdomen increases the food intake and body mass of anorexia model rats,reduces the residue of gastric contents,improves the small intestine propulsion rate,and therefore has a good therapeutic effect on anorexia.The mechanism may be related to inhibiting the secretion of CCK-8 and NT in plasma and promoting the release of MTL and GAS in serum.
4.Influence of Tuina plus oxiracetam on serum inflammatory factors and oxidative stress in mild vascular dementia patients
Nan GU ; Yuexian FAN ; Ling ZHOU ; Yan ZHANG ; Weijie BAI ; Yanfeng LI ; Liyun YANG ; Jinlong LI ; Jue HONG
Journal of Acupuncture and Tuina Science 2022;20(4):273-280
Objective: To observe the clinical efficacy of Tuina (Chinese therapeutic massage) plus oxiracetam in treating mild vascular dementia (VD) and seek its underlying mechanism. Methods: Ninety-six patients with mild VD were randomized into an observation group and a control group, with 47 cases in the observation group and 49 cases in the control group. The control group received oral oxiracetam capsules for treatment, and the observation group was given additional Tuina treatment. Before and after treatment, the mini-mental state examination (MMSE) was adopted to assess the patient's cognitive function; the activities of daily living (ADL) scale was used to evaluate their ability to conduct daily activities; changes in the serum inflammatory factors and oxidative stress indicators were also detected. Results: After treatment, the serum content of malondialdehyde (MDA) decreased in both groups (P<0.05) and was lower in the observation group than in the control group (P<0.05); the serum contents of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) increased in both groups (P<0.05) and were higher in the observation group than in the control group (P<0.05); the serum contents of interleukin (IL)-1, tumor necrosis factor (TNF)-α, IL-6, and IL-8 declined in both groups (P<0.05) and were lower in the observation group than in the control group (P<0.05). After the intervention, the levels of systolic velocity (Vs) and mean velocity (Vm) of the middle cerebral artery elevated, and the pulsatility index (PI) dropped in patients in the two groups, showing significant intra-group differences (P<0.05); the levels of Vs and Vm in the observation group were higher than those in the control group, and the PI was lower in the observation group than in the control group, showing significant between-group differences (P<0.05). The MMSE and ADL scores increased in both groups after the intervention (P<0.05) and were higher in the observation group than in the control group (P<0.05). Conclusion: In the treatment of mild VD, Tuina plus oxiracetam can improve the cerebral blood supply, ADL, and cognitive function; the mechanism may be associated with the reduction of oxidative stress damages and inflammatory reactions.
5.Evaluation on Scales Based on Unidimensional Item Response Models
Wenjing BAI ; Liyun HE ; Yanhong ZHANG ; Weimin LIU ; Baoyan LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2017;19(9):1473-1480
This study was aimed to assess scales with the unidimensional item response models (UIRM) and classical test theory.The reliability and validity of 841 sub-health status samples were analyzed by CTT.Surveying evaluation from the project perspective was also conducted.The results showed that Cronbach'so coefficients in five areas were all larger than 0.6 with relatively high consistency.The scale structures of five areas were relatively good and the CFI was 0.96.When discriminations of all items were larger than 0.3,the sensitivity of item for its area was high.When the scale test information was 33.8,the accuracy of sub-health status was relatively high.However,there were still some items should be further revised.It was concluded that UIRM can be used to evaluate the psychological measurement performance in the project.It is a supplement to the evaluation result of classical test theory.The reliability and validity of sub-health status scale are relatively high.After IRT correction,it can be used for human-computer interaction techniques “preventive treatment” dynamic monitoring.
6.Three-Dimensional Finite Element Analysis of PEEK Customized Reconstruction Plate for Mandibular Defect
Hui GAO ; Liyun BAI ; Xian LI ; Ping JI ; Chao WANG
Journal of Medical Biomechanics 2019;34(2):E193-E199
Objective To study the stress distributions of mandible defect by reconstruction with polyetheretherketone (PEEK) and its composite reconstruction plate through three-dimensional finite element analysis. Methods The finite element models of reconstruction plate of titanium alloy, PEEK, carbon-fiber-reinforced polyetheretherketone (CFR)-PEEK with 30% endless carbon fibers and CFR-PEEK with 68% endless carbon fibers were established by CBCT scanning,Mimics software,SolidWorks, Geomagic Studio and ANSYS Workbench software, and titanium alloy served as control. Two occlusal situations were simulated in the mandible model. Loading I: anterior region loading with 300 N; loading II: left posterior region with 300 N. Results The ratio of the maximum Von Mises stress of the reconstructed plate to its yield strength under two load situations: PEEK system > 30%CFR-PEEK system > titanium alloy system > 68% CFR-PEEK system; the maximum stress of the mandible: PEEK system > 30% CFR-PEEK system > titanium alloy system > 68% CFR-PEEK system. In the PEEK system, the maximum Von Mises stress of reconstructed plate and mandibular exceeded its yield strength under loading I; in the other systems, the maximum Von Mises stresses of mandible and reconstruction plate were below the yield strength. Conclusions The reconstruction plate of CFR-PEEK with 68% endless carbon fibers distributed the stresses in a similar manner as the titanium reconstruction plate, which could meet the mechanical strength requirements of mandible defect reconstruction. The fracture risk of the reconstructed plate was lower than that of titanium alloy, but the incidence of stress shielding was slightly higher. The result can provide references for the selection of materials and clinical application of reconstruction plate for mandibular defects.
7.Immunogenicity of the truncated NDV F protein surface-displayed on Lactobacillus casei.
Huanhuan LIU ; Shudong LI ; Yuqing YANG ; Xiaoying SUN ; Yan LI ; Xinyang LIU ; Xiaoyan CHEN ; Lianmei ZHANG ; Yongfei BAI ; Xilin HOU ; Liyun YU
Chinese Journal of Biotechnology 2019;35(8):1453-1462
To evaluate immune efficacy of the recombinant Lactobacillus casei, we constructed pLA-Newcastle disease virus (NDV)-F/L. casei and obtained the expression products. PCR amplified the NDV F gene carrying part of the major epitopes. The target gene was inserted to the shuttle plasmid pLA, and then transformed into Escherichia coli BL21 (DE3) in order to screen positive recombinant plasmid. The positive recombinant plasmid was transformed into L. casei by electroporation to construct pLA-NDV-F/L. casei. The positive strains were identified by PCR. The reactivity of the recombinant bacteria was identified by Western blotting and the protein expression was detected by indirect immunofluorescence, flow cytometry and laser confocal microscopy. The 14-day-old chickens in each group were vaccinated by oral plus nose drops. The pLA-NDV-F/L. casei twice immunization group and three times immunization group, the commercial vaccine group, the pLA/L. casei group, the unchallenge PBS and the challenge PBS group were established. IgG in serum and sIgA in the lavage fluid of intestinal, nasal and lung were detected by ELISA. The protection rate of chickens was evaluated. The results showed that 94.10% of the recombinant bacteria expressed the F protein. The recombinant protein was highly expressed on the surface of L. casei with a protein size of 62 kDa, which specifically bound to anti-NDV serum. The levels of anti-F IgG and sIgA antibodies in each test group were significantly higher than those in the control groups. The duration of antibody in the pLA-NDV-F/L. casei three-time immunization group lasted 28 days longer than that in the twice immunized group, and there was no significant difference between antibody peak values. The attack protection rates in each group of immunized pLA-NDV-F/L. casei three times, twice, attenuated vaccine, pLA/L. casei and PBS were 80%, 80%, 90%, 0% and 0%, respectively. Therefore, the antigenic protein of NDV F was successfully expressed by L. casei expression system, which has of reactogenicity and immunogenicity, and could induce protective immune responses in chickens.
Animals
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Antibodies, Viral
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Chickens
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Immunization
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Lactobacillus casei
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Newcastle disease virus
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Vaccines, Attenuated
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Viral Vaccines
8.Establishing the acupuncture-moxibustion clinical trial registry and improving the transparence of clinical trials of acupuncture and moxibustion.
Yali LIU ; Liyun HE ; Jia LIU ; Xingyue YANG ; Dongning YAN ; Xin WANG ; Lin LUO ; Hongjiao LI ; Shiyan YAN ; Tiancai WEN ; Wenjing BAI ; Taixiang WU ; Baoyan LIU
Chinese Acupuncture & Moxibustion 2017;37(7):685-689
As a kind of intervention measures of traditional Chinese medicine, acupuncture-moxibustion is highly adopted on global clinical practice. Even though the global clinical trial registration system was established more than 10 years ago, the proportion of acupuncture-moxibustion clinical trial registration is still very low; and it is very problematic on the methodological quality and report quality in the published acupuncture-moxibustion clinical trials. In order to manage particularly the acupuncture-moxibustion clinical trials, China Academy of Chinese Medical Sciences, collaborated with China Association of Acupuncture and Moxibustion and World Federation of Acupuncture Societies, established the Acupuncture-Moxibustion Clinical Trail Registry (AMCTR). AMCTR is a secondary registry platform affiliated to the Chinese Clinical Trial Registry (ChiCTR) and WHO International Clinical Trials Registry Platform (ICTRP), specifically for the acceptance and management of clinical trials in the field of acupuncture and moxibustion. It is a nonprofit academic organization, located in China Academy of Chinese Medical Sciences.