Biomacromolecules play an important role in the treatment of many diseases, but as a result of cell membrane serving as the natural barriers, only the small molecular compounds whose molecular weights are smaller than 600 Da can get through cell membrane and enter the cell. In recent years, some short peptides (the length less than 30 amino acids) are found to have the cell-penetrating function, called cell-penetrating peptides (CPPs). They are able to effectively translocate segments of protein, polypeptides, nucleic acid into the cells of many mammal animals with many methods. They have high transduction efficiency and will not lead to cell damage. So, the discovery of CPPs has a very good applicable prospect in such research fields as cell-biology, gene-therapy, drug transduction in vivo, evaluation of clinical medicine and medical immunology. This paper reviews the types and characteristics of CPPs, internalization mechanisms, applications, and their existing problems.

Biomacromolecules play an important role in the treatment of many diseases, but as a result of cell membrane serving as the natural barriers, only the small molecular compounds whose molecular weights are smaller than 600 Da can get through cell membrane and enter the cell. In recent years, some short peptides (the length less than 30 amino acids) are found to have the cell-penetrating function, called cell-penetrating peptides (CPPs). They are able to effectively translocate segments of protein, polypeptides, nucleic acid into the cells of many mammal animals with many methods. They have high transduction efficiency and will not lead to cell damage. So, the discovery of CPPs has a very good applicable prospect in such research fields as cell-biology, gene-therapy, drug transduction in vivo, evaluation of clinical medicine and medical immunology. This paper reviews the types and characteristics of CPPs, internalization mechanisms, applications, and their existing problems.

Objective To evaluate the role of brain-derived neurotrophic factor (BDNF) in inflammatory pain in rats. Methods Sixty female SD rats weighing 150-180 g in which intrathecal (IT) catheters were successfully placed without complication were randomly divided into 5 groups (n= 12 each): group Ⅰ sham operation; group Ⅱ sham operation + IT anti-BDNF antibody; group Ⅲ inflammatory pain; group Ⅳinflammatory pain + IT control IgG and group Ⅴ inflammatory pain + IT anti-BDNF antibody. Inflammatory pain was induced by injecting complete Freund's adjuvant (CFA) into ankle joint cavity of left hindpaw, while in sham operation group equal volume of normal saline was injected instead of CFA. Anti-BDNF antibody or control IgG 15 μg/10 μl was injected IT once a day for 3 days after inflammatory pain was induced. Paw withdrawal latency to thermal stimuli (PWTL) was measured one day before and at 3, 5, 7, 10 and 14 d after inflammatory pain was induced. The rat was sacrificed on 3 rd day of IT anti-BDNF antibody or control IgG injection. The lumbar segment L4-6 of the spinal cord was removed for detection of the expression of BDNF and p-ERK1/2 by immunohistochemistory and Western blot. Results Intra-articular CFA injection significantly increased the expression of BDNF and p-ERK1/2 in the spinal cord in group Ⅲ as compared with sham-operated animals in group Ⅰ . IT antiBDNF antibody injection significantly suppressed the expression of BDNF and p-ERK1/2. PWTL was significantly shortened after intra-articular CFA injection in group Ⅲ as compared with group Ⅰ . IT anti-BDNF antibody reversed the inflammation-induced thermal hyperalgesia in group Ⅴ but IT control IgG did not. Conclusion BDNF in the spinal cord may be involved in inflammatory pain through p-ERK1/2 signal transduction pathway.

Objective To explore the effects of different dose rates of X-ray under the same dose on cell clonogenic formation in non-small-cell lung cancer cell line A549 in order to provide experimental basis for clinical radiotherapy plan. Methods The A549 cells were cultured at low density and irradiated with X-rays at dose of 4 Gy and selected dose rates of 1, 2, 4 and 6 Gy/min, respectively, from a linear accelerator. The 8th day after irradiation, the cells were fixed and stained with Giemsa solution, and colonies containing at least 50 cells were counted. The plating efficiency and surviving fraction were calculated. Results The clonogenic number in non-irradiated cells was 88.6±4.6. The numbers were significantly reduced in irradiated cells at dose rate 1, 2, 4 and 6 Gy/min (12.3±3.4, 9.0±0.8, 5.6±1.0, 11.5±1.7, respectively) than that in non-irradiated control cells (F=678.799, P<0.05). The plating efficiencies were decreased in irradiated cells, especially in 4 Gy/min irradiated cells, which was lower than that in any of the other three dose rate groups (P< 0.05). Conclusions Though at same radiation dose, cancer cells have different clonogenic formation efficiency when irradiation with X-ray at different dose rates. Thus, treatment with optimal dose rate may improve the radiotherapy efficacy.

AIM: To study the effects of apolipoprotein (apo) A-Ⅰon ATP binding cassette transporter A1 (ABCA1) degradation and cholesterol efflux in THP-1 macrophage-derived foam cells. METHODS: After exposure of the cultured THP-1 macrophage-derived foam cells to apolipoprotein A-Ⅰ for different time, cholesterol efflux, ABCA1 mRNA and protein level were determined by liquid scintillation counting, reverse transcriptase-polymerase chain reaction and Western blotting, respectively. The mean ABCA1 fluorescence intensity on THP-1 macrophage-derived foam cells was detected by flow cytometry. RESULTS: ApoA-Ⅰ markedly increased ABCA1-mediated cholesterol efflux from THP-1 macrophage-derived foam cells. This was accompanied by an increase in the content of ABCA1. ApoA-Ⅰ did not alter ABCA1 mRNA abundance. Thiol protease inhibitors increased the level of ABCA1 protein and slowed its decay in THP-1 macrophage-derived foam cells, whereas none of the proteosome-specific inhibitor lactacystin, other protease inhibitors, or the lysosomal inhibitor NH_4Cl showed such effects. The apoA-Ⅰ mediated cellular cholesterol efflux was enhanced by thiol protease inhibitors. CONCLUSION: Thiol protease inhibitors might provide an alternative way to upregulate ABCA1 protein. This strategy is especially appealing since it may mimic the stabilizing effect of the natural ligands apoA-Ⅰ.

OBJECTIVETo study the feasibility of intraoperative radiotherapy (IORT) in primary liver cancer.

METHODSBased on the target of dose curves, the dose-volume histogram (DVH) and cost of radiation equipment and radiation therapy, IORT was compared with protonbeam therapy (PBT) and 3DCRT in 16 patients with primary liver cancer using the therapy plan system (TPS).

RESULTSIORT had significantly better performance than 3DCRT to allow a target region surrounded by 90% of the dose lines. IORT was similar to protonbeam therapy in terms of target region surroundings and absorbed dose in the normal organs, but the cost of IORT was significantly lower.

CONCLUSIONThe TPS of IORT is better than 3DCRT and similar to protonbeam therapy in the treatment of primary liver cancer with similar cost to 3DCRT. IORT can effectively protect the neighboring sensitive organs and improve the absorbed dose in the tumors and the local control rate.

Aged ; Feasibility Studies ; Female ; Humans ; Intraoperative Care ; Liver Neoplasms ; radiotherapy ; surgery ; Male ; Middle Aged ; Radiotherapy ; methods ; Radiotherapy Dosage ; Radiotherapy, Adjuvant ; methods

Aim To investigate the mechanism for the inhibitory effect of hydrogen sulfide on the expression of tissue factor(TF)induced by oxidative low-density lipoprotein(ox-LDL)in endothelial cells.Methods Human umbilical vein endothelial cells (HUVECs ) were treated with 50 mg·L-1 ox-LDL in the absence or presence of different concentrations of NaHS (25 , 50,100 and 200 μmol·L-1 )for 24 h.The mRNA expression and protein content of TF in HUVECs were determined by reverse transcription PCR and ELISA, respectively.The content of intracellular reactive oxy-gen species (ROS)was determined by DCFH,an oxi-dative sensitive fluorescent indicator.The activation of nuclear factor-kappaB (NF-κB)was estimated by its expression in nuclear extracts analyzed by Western blot.Results Ox-LDL induced TF mRNA expression and increased TF protein content in HUVECs.The in-crease in intracellular ROS production and the activa-tion of NF-κB were observed in HUVECs treated with ox-LDL.However,NaHS could markedly inhibit the increases in TF mRNA and protein levels induced by ox-LDL.Also the elevation of intracellular ROS pro-duction and the activation of NF-κB elicited by ox-LDL were significantly suppressed by pretreatment with NaHS.In addition,pretreatment with BAY 1 1-7082 (10 μmol·L-1 ),the inhibitor of NF-κB or N-acetyl-L-cysteine(1 mmol·L-1 ),an antioxidant,could also decrease the TF mRNA and protein level as well as ROS production and NF-κB activation induced by ox-LDL in HUVECs,similar to the effects of 200 μmol· L-1 NaHS.Conclusion The mechanism for the in-hibitory effect of H2 S on the ox-LDL- induced TF ex-pression in endothelial cells may be related to inhibi-ting intracellular ROS production and subsequently NF-κB activation.

Objective To study the effects of selenium deficiency,iodine deficiency and combined selenium and iodine deficiency on bone and cartilage growth in the parental and the first filial generation rats. Methods Forty-eight weanling healthy SD rats were randomly divided into selenium deficieney, iodine deficiency, combined selenium and iodine deficiency and control groups according to their body mass. These rats were fed with selenium deficiency, iodine deficiency, combined selenium and iodine deficiency, and normal fodder, respectively. The parental rats (about 3 months old) were mated in each group 8 weeks after the beginning of the experiment. Right tibias and left knee joints were collected when the parental generation rats were about 6 months and the first filial generation rats were about 3 months old. Tibial length, mid-shaft tibial diameter, and articular cartilage diameters of the right tibias were measured by vernier caliper. Left knee joints were embedded and cut into sections and the thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes in growth plate cartilage were observed under the light microscope. Results The selenium deficiency had significant effect on serum selenium level of the parental and the first filial generation rats(F value were 239.56,232.68, P< 0.01), and also on serum T4 level of the first filial generation rats(F value were 6.95, P < 0.05). The iodine deficiency had significant effect on serum T3 and T4 level in the two generations rats(F value were 14.11,14.05,30.29,34.53, P < 0.01 ). There were interactions between selenium deficiency and iodine deficiency on serum T4 level in the first filial generation rats (F= 5.99, P< 0.05). The serum selenium of selenium deficiency group[ (30.28 ± 6.34), (43.95 ± 9.75)μg/L],combined selenium and iodine deficiency group[ (30.33 ± 5.18), (35.40 ± 3.16)μg/L] were significantly lower than iodine deficiency group[(345.83 ± 29.55), (245.24 ± 9.95)μg/L] and the controls[ (358.64 ± 30.50), (236.50 ±9.75) μg/L] in the two generations. The serum T3 of combined selenium and iodine deficiency group [(0.55 ± 0.05 ),(0.88 ± 0.14)nmol/L] were significantly lower than the controls[(0.75 ± 0.08), (1.26 ± 0.26)nmol/L] in the two generations. The serum T4 of iodine deficiency [ (24.11 ± 2.29), (42.10 ± 8.92) nmol/L ] and combined selenium and iodine deficiency group[ (20.66 ± 1.93), (26.55 ± 5.98)nmol/L] were significantly lower than the controls[ (36.15 ±2.74), (52.79 ± 8.84)nmol/L] and selenium deficiency group[ (28.12 ± 3.33), (52.02 ± ll.99)nmol/L] in the two generations. The selenium deficiency and iodine deficiency had significant effect on tibial length, thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes in first filial generation rats(F values were 24.31,6.98,40.76,56.15,25.24,82.82, 10.07,5.57, P <0.05 or <0.01). There were interactions between selenium deficiency and iodine deficiency on tibial length, thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes (F values were 5.68,24.86,41.82,9.12, P <0.05 or <0.01 ). The tibial length of the selenium deficiency group[ (33.17 ± 0.34)mm] and combined selenium and iodine deficiency group[ (31.30 ± 0.87)mm] were significantly lower than the controls[ (34.12 ± 0.32)mm, P< 0.05]. Thickness of the growth plate cartilage [ (1.60 ± 0.18)mm ], layers of proliferative chondrocyte (8.54 ± 0.81), and hypertrophic chondrocyte (4.95 ± 0.37)of the combined selenium and iodine deficiency group were significantly decreased when compared to the selenium deficiency group[ (3.03 ± 0.10)mm, 14.68 ± 0.84,6.60 ± 0.31], iodine deficiency group[ (2.90 ± 0.09)mm, 13.75 ±0.33,6.61 ± 0.84 ] and the controls [ (3.19 ± 0.09) mm, 14.94 ± 0.36, 6.64 ± 0.26, P <0.05]. Thickness of the growth plate cartilage, layers of proliferative chondrocyte of the iodine deficiency group were lower than the controls(P<0.05). Conclusions Selenium deficiency impair tibial growth in first filial generation rats, iodine deficiency retarded the chondroncyte proliferation and decreases the thickness of growth plate cartilage in first filial generation rats, and combined selenium and iodine deficiency significantly impair the growth of bone and cartilage in first filial generation rats.

OBJECTIVETo identify whether GC-box (-348 to -338) in human insulin gene promoter is a key cis-acting element.

METHODSHuman insulin gene promoter was sub-cloned into secreted alkaline phosphatase (SEAP) reporter plasmid. The deletion and mutation of GC-box in insulin gene promoter was performed. The activity of human insulin gene promoter was determined by evaluating the activity of SEAP in the supernatant of cell culture after the reporter plasmids were transfected in beta cell line betaTC3.

RESULTDeletion and mutation of GC box in human insulin gene promoter did not result in significant changes of the activity of the promoter in betaTC3.

CONCLUSIONThe GC-box is not a key cis-acting element in human insulin gene promoter.

Alkaline Phosphatase ; genetics ; metabolism ; Base Sequence ; Cell Line ; Enhancer Elements, Genetic ; GC Rich Sequence ; Gene Expression Regulation ; Humans ; Insulin ; genetics ; Promoter Regions, Genetic ; Recombinant Fusion Proteins ; genetics ; metabolism ; Sequence Deletion ; Transcription, Genetic ; Transfection

Objective To study the prevention effects of AduoLa Fuzhenglin(ADL)Oll the brain injury induced by microwave radiation in rats.Methods A total of 140 male Wismr rats were divided randomly into 5 groups,including control group,microwave exposed group,low dosage(0.75 g·kg-1·d-1)group.middle dosage(1.5 g·kg-1·d-1)group and high dosage(3 g·kg-1·d-1)group.Rats in three ADL groups were lavaged with ADL per day for 2 weeks before radiation.After administration,rats were exposed to microwave at 30 mW/cm2 for 15 min.The abilities of learning and memory were detected by Morris water maze,and the contents of amino acids neurotransmitter of hippocampus were detected by HPLC, then the histology and uhrastrncture of hippocampus were observed with light and electron microscope at 6 h,7 and 14 d after exposure.Results The abilities of learning and memory were declined(F=0.000-0.043,P<0.05)from 6 h to 7 d after exposure,and the contents of four kinds of amino acid neurotransmitter in hippocampus were decreased,of which GLU,GLY and GABA were decreased significantly(F=0.000-0.007,P<0.01)at 6h after exposure,then tissue edema,neuronal degeneration,neuron mitoehondria swelling and cavitation,endocytoplasmie rotieulum broaden,synaptic cleft blurred,and perivascular space widen were found in the hippocampus at 6 h and 7 d after exposure.The changes in low dosage group were similar to those of the radiation group.However,in middle and high dosage groups,the abilities of learning and memory were normal to some extent with the significant differences compared to the radiation group from 6 h to 7 d after exposure(F=0.015-0.028.P<0.05).The contents of four kinds of amino acid neurotransmitter were not decreased,especially GLU contents close tO normal level.There were significant differences between middle and high dosage groups and radiation group at 6 h after exposure(F=0.000-0.042,P<0.05).Moreover,no obvious injury in the hippocampus was observed in middle and high dosage groups at 6 h and 7 d after exposure.Conclusions Exposure to 30 mW/cm2 microwave radiation could decrease the abilities of learning and memory,induce amino acid neurotransmitter turbulence,and injure the histology and uhrastructure of hippocampus.ADL at the dosages of 1.5 and 3 g·kg-1·d-1 would have preventive effects on the injury induced by microwave exposure.The concentration of 1.5 g·kg-1 ·d-1 of ADL might be the effective dosage to prevent the brain damage after microwave exposure.