Cell-penetrating peptides (CPPs) offer a non-selective and receptor-independent mode to promote cellular uptake. Although the non-specificity of CPP-mediated internalization allows this approach applicable to a wide range of tumor types potentially, their universality is a significant obstacle to their clinical utility for targeted delivery of cancer therapeutics and imaging agents. Accordingly, many reports have focused on selective switching of systemically delivered inert CPPs into their active form in lesions (tumor). In this review, our attention is mainly confined to such an enzyme-sensitive domain incorporated delivery system with activatable CPPs (ACPPs), which have displayed the exciting strength in balancing the CPPs' pros and cons, and potential in the treatment and diagnosis of some diseases.
Cell-Penetrating Peptides ; chemistry ; Drug Delivery Systems ; Enzymes ; chemistry ; Humans ; Neoplasms ; drug therapy

Objective To comprehensively study the oxidative stress of bone tissue in rats with chronic fluorosis treated with anti-oxidant,the oxidative damage of lipid,protein and DNA.Methods Forty Wistar rats weaned 2 weeks were randomized by weight and divided into 4 groups according to body weight,control group (treated with tap water) and 3 NaF (sodium fluoride) exposure groups (treated with NaF at 50,150 and 250 mg/L),5 female rats and 5 male rats in each group.NaF was given through drinking water.After 6 months of treatment,a 12-hour urine samples were collected,then rats were killed,serum was collected,right rear tibiofibula was separated.Bone and urinary fluoride content and incidence rate of dental fluorine were studied and the levels of bone tissue suppression function of hydroxy free radical,superoxide dismutase (SOD),catalase (CAT),glutathione peroxidase (GSH-Px),8-hydroxydeoxyguanosine (8-OHdG),protein carbonyls (PCO),and malonaldehyde (MDA) were assayed.Results ① Results of suppression function of hydroxy free radical:The difference of bone tissue suppression function of hydroxy free radical among control [(22.99 ± 4.31)U/mg prot],low-excess dose [(22.76 ± 8.11)U/mg prot],medium-excess dose [(13.47 ± 4.56)U/mg prot] and high-excess dose [(19.40 ± 5.92)U/mg prot] groups was statistically significant (F =5.01,P ＜0.05).②Results of SOD:The difference of bone tissue SOD among control [(5.06 ± 1.16)U/mg prot],low-excess dose [(5.32 ± 1.18)U/mg prot],medium-excess dose [(3.71 ± 0.72)U/mg prot] and high-excess dose [(4.80 ± 1.10)U/mg prot] groups was statistically significant (F =4.44,P ＜0.05).③ Results of CAT:The difference of bone tissue CAT among control [(25.20 ± 5.91)U/mg prot],low-excess dose [(22.53 ± 7.10) U/mg prot],medium-excess dose [(17.96 ± 4.71)U/mg prot] and high-excess dose [(19.52 ± 5.52)U/ mg prot] groups was statistically significant (F =2.85,P ＜0.05).④Results of GSH-Px:The differences of bone tissue GSH-Px among control [(52.86 ± 12.88)U/mg prot],low-excess dose [(70.05 ± 15.72)U/mg prot],medium-excess dose [(51.55 ± 6.97)U/mg prot] and high-excess dose [(57.47 ± 10.99) U/mg prot] groups was statistically significant (F =4.89,P ＜0.05).⑤Results of PCO:The differences of bone tissue PCO among control [(58.73 ± 20.86)ng/L],low-excess dose [(89.41 ± 26.20)ng/L],medium-excess dose [(97.07 ± 22.24)ng/L] and highexcess dose [(83.96 ± 29.55)ng/L] groups was statistically significant (F =4.43,P ＜0.05).⑥Results of 8-OHdG:The differences of bone tissue 8-OHdG among control [(87.66 ± 6.32)ng/L],low-excess dose [(86.31± 6.30)ng/L],medium-excess dose [(92.17 ± 4.28)ng/L] and high-excess dose [(88.02 ± 6.14)ng/L] groups was not statistically significant (F =1.88,P ＞ 0.05).⑦Results of MDA:The differences of bone tissue MDA among control [(3.70 ± 1.73) nmol/mg prot],low-excess dose [(2.10 ± 0.95)nmol/mg prot],medium-excess dose [(3.32± 2.20)nmol/mg prot] and high-excess dose [(2.71 ± 2.18)nmol/mg prot] groups was not statistically significant (F =1.37,P ＞ 0.05).Conclusions The activity of SOD and CAT of bone tissue are inhibited and suppression function of hydroxy free radical is decreasing under fluorosis influence,which results in protein damage.Oxidative stress is considered to be one of the mechanisms of skeletal fluorosis.

OBJECTIVETo compare the therapeutic effect of Astragalus and Angelica Mixture (AAM) on treating CKD patients according to different CKD primary diseases, staging and TCM syndromes.

METHODSA multicentre, open-label, and self control clinical design was used, and thirty-two patients in line with inclusive criteria were recruited. Based on maintaining their previous basic CKD treatment, patients additionally took AAM (Astragalus and Angelica each 30 g), once a day, three months consisted of one therapeutic course. Serum creatinine (SCr), estimated glomerular filtration rate (eG- FR), 24 h urinary total protein (UTP), plasma albumin (ALB), hemoglobin (Hb), and changes of TCM syndrome factor integrals were compared before treatment, at the end of month 1, 2, and 3. The differences in the aforesaid indices were compared between CKD patients with different CKD primary diseases (chronic glomerulonephritis, chronic renal tubulointerstitial disease, hypertensive renal damage), different CKD stages (CKD 3 and CKD 4), and patients of qi-blood deficiency syndrome (QBDS) and non-QBDS.

RESULTSAAM could improve 78.12% (25/32) patients' renal function. Compared with before treatment, SCr decreased (12.08% +/- 10.11%), eGFR increased (21.14% +/- 18.55%), and ALB increased (2.76% +/- 1.97%) at the end of 3-month treatment (all P < 0.05). As for TCM syndrome factor integrals, compared with before treatment, the integrals for qi deficiency syndrome, blood deficiency syndrome, and yin deficiency syndrome decreased, while the integrals for dampness heat syndrome and turbid-toxin syndrome increased (all P < 0.05). There was no obvious difference in all indices except the integral for hypertensive renal damage patients of yin deficiency syndrome (P > 0.05). The SCr decreasing percent was 19.82% +/- 8.30% for patients of non-QBDS and 5.24% +/- 10.75% for patients of QBDS. The latter was higher with statistical difference (P < 0.05). As for TCM syndrome factor integrals, the integral differences of qi deficiency and blood deficiency were obviously higher in patients of QBDS, when compared with patients of non-QBDS (P < 0.05).

CONCLUSIONAAM could improve the renal function of CKD patients, elevate their ALB levels, and ameliorate associated qi deficiency syndrome, blood deficiency syndrome, and yin deficiency syndrome, especially for CKD patients of QBDS.

Adolescent ; Adult ; Aged ; Angelica ; Astragalus Plant ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Phytotherapy ; Renal Insufficiency, Chronic ; drug therapy ; Treatment Outcome ; Yin Deficiency ; drug therapy ; Young Adult

OBJECTIVE To investigate enhanced immune function of methionine encephalin (MENK) and its anti-tumor mechanism in CT26 colon cancer mouse model. METHODS 3×106 CT26 cells were implanted subcutaneously in BALB/c mice. Four days after, MENK was peritoneally administrated at the concentration of 20 mg·kg-1 for 14 d. The percentage of MDSCs in bone marrow, spleen, blood, tumor and liver were detected by flow cytometry. Non- esterified fatty acid (NEFA), triglycerides (TG) and total cholesterol (T-CHO) in liver homogenate were tested by a NEFA test kit, a TG test kit and a T- CHO test kit respectively. qRT- PCR and Western blot were used to measure mRNA and protein levels of inflammation-, glycometabolsim- and lipometabolsim-associated indexes in liver. RESULTS MENK decreased percentages of MDSCs in bone marrow, spleen, blood and tumor in colon cancer mice. MENK-treated mice displayed elevated ratio of CD4+T and CD8+T cells in spleen as well as increased T and B lymphocytes proliferation. Meanwhile, MENK also ameliorated liver damage reflected by lower levels of GPT and GOT in serum and reduced risks of cancer- associated index including inflammation, high lipid and high glucose. Furthermore, MENK lowered down the levels of NEFA, TG and T- CHO in liver homogenate. MENK treatment decreased expression of p- STAT3, increased expression of p-AKT, IRS1 and Glut4 at protein level as well as reduced lipogenesis-associated genes and elevated glycolysis-associated genes in liver of tumor bearing mice. Also, abated expression of genes associated with MDSCs generation (M-CSF, GM-CSF, IL-6, IL-1β) and migration (S100A9, KC) was observed within shrunken subcutaneous tumor by MENK intervention. CONCLUSION MENK has the ability to strength immune function against colon cancer by reducing MDSCs and improving liver metabolism.

Objective To explore the expression of HCK gene during the cardiomyocyte differentiation of mouse embryonic stem cells and analyze the role of HCK gene in maintenance of pluripotency of embryonic stem cells.Methods Mouse embryonic stem cells were cultured,then induced to differentiate into cardiomyocytes.Total RNAs were isolated from mouse embryonic stem cells in the differentiation days:0 day(D0),the second day(D2),the fourth day(D4),the sixth day(D6),the eighth day(D8),respectively.The levels of HCK mRNAs were assessed by the method of semi-quantitive reverse transcriptase-polymerase chain reaction(RT-PCR).In the meanwhile,Total proteins were also isolated from mouse embryonic stem cells in the differentiation D0,D2,D4,D6,D8,and the levels of HCK proteins were evaluated by Western-blot.Results HCK mRNAs could be detected in the mouse embryonic stem cells in D0 and D2,however,they were undetectable from D4 to D8.The expression of HCK mRNAs was rapidly down-regulated during cardiomyocyte differentiation of mouse embryonic stem cells.Expression of HCK proteins,which coincided with HCK mRNAs,down-regulated during differentiation and couldn't be detected in D4.Conclusions With the cardiomyocyte differentiation of mouse embryonic stem cells,the expression of HCK in the levels of mRNA and proteins are sharply down-regulated;HCK may play an important role in maintaining the pluripotency of embryonic stem cell.

Adult ; Cloning, Molecular ; Female ; Gene Expression ; Humans ; Liver Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Oncogenes

Objective To evaluate the affection of length of indwelling gastric tube in stroke patients with dysphagia.Methods 78 stroke patients with dysphagia were assigned into two groups randomly,39 cases in each group.tube length was 55-65 cm in observation group,45-55 cm in control group.Results 3 days and 1 week after intubation,the incidence rate of tube pulling out,gastrointestinal complications such as nausea and vomiting,aspiration,diarrhea and abdominal distension,upper gastrointestinal hemorrhage of observation group were lower than control group(P<0.05);2 weeks after intubation,the aspiration pneumonia ineidence of observation group Was lower than control group,the difference was statistically significant(P<0.05).Conclusions The incidence of vomiting,reflux,diarrhea and abdominal distension,upper gastrointesdnal hemorrhage,pneumonia decreased remarkablely.

OBJECTIVETo explore the mechanism of cake-separated moxibustion in treatment of hyperlipemia.

METHODSSixty rabbits were randomly divided into 4 groups, a blank group,a model group, a direct moxibustion group and a cake-separated moxibustion group. Hyperlipemia model was developed by high fat diet of cholesterol. Changes of ultrastructures of endothelial cells of the aorta of the rabbit were observed with electron microscope.

RESULTSThe endothelial cells in the cake-separated moxibustion group were more intact, most of them were normal in forms, internal elastic membrane was continuous, their thickness was even, the cells of smooth muscles in the medial membrane were relatively normal, which are similar to those in the blank control group. But the structure of endothelial cells of the aorta in the model group disappeared, in cytoplasm the sedimentation of a great number of lipids can be seen, internal elastic membrane was interrupted, the thickness was uneven, with focal dissolution, the cells of smooth muscle in the medial membrane had sedimentation of lipids, with frothy change.

CONCLUSIONCake-separated moxibustion has a certain protective action on endothelial cells of the aorta in the rabbit of hyperlipemia.

Animals ; Aorta ; Endothelial Cells ; Hyperlipidemias ; Lipids ; Moxibustion ; Rabbits

OBJECTIVETo study the inhibitory effect of paeoniflorin (PAE) on TNF-α-induced TNF receptor type I (TNFR1)-mediated signaling pathway in mouse renal arterial endothelial cells (AECs) and to explore its underlying molecular mechanisms.

METHODSMouse AECs were cultured in vitro and then they were treated by different concentrations PAE or TNF-α for various time periods. Expression levels of intercellular cell adhesion molecule-1 (ICAM-1) were detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 6-h TNF-α 30 ng/mL), the low dose PAE group (cultured by 2-h PAE 0.8 μmo/L plus 6-h TNF-α 30 ng/mL), the middle dose PAE group (cultured by 2-h PAE 8 μmol/L plus 6-h TNF-α 30 ng/mL), the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 6-h TNF-α 30 ng/mL) with Western blot analysis. Nuclear translocation of transcription factor NF-κB (NE-κB) was detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 45-mm TNF-α 30 ng/mL), and the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 45-min TNF-α 30 ng/mL) by immunofluorescent staining. Expression levels of the phosphorylation of extracellular signal-regulated (protein) kinase (ph-ERK) and p38 (ph- p38) were detected in the normal group (cultured by serum-free culture media) and the high dose PAE group (2-h PAE 80 μmol/L culture) by Western blot. NF-κB inhibitor-α (IκBα) protein expressions were detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 30-min TNF-α 30 ng/mL), the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 30-min TNF-α 30 ng/mL), the p38 inhibitor group (SB group, pretreatment with SB238025 25 μmol/L for 30 min, then treated by PAE 80 μmol/L for 2 h, and finally treated by TNF-α 30 ng/mL for 30 min), the ERK inhibitor group (PD group, treated by PD98059 50 μmol/L for 30 min, then treated by PAE 80 μmol/L for 2 h, and finally treated by TNF-α 30 ng/mL for 30 min) by Western blot.

RESULTSCompared with the normal group, ICAM-1 protein expression levels obviously increased (P < 0.01). Compared with the TNFα group, ICAM-1 protein expression levels were obviously inhibited in the high dose PAE group (P < 0.05). Protein expression levels of ph-p38 and ph-ERK were obviously higher in the hIgh dose PAE group (P < 0.05). Compared with the normal group, IκBα protein expression levels obviously decreased in the TNF-α group (P < 0.01). Compared with the TNFα group, TNF-α-induced IκBα degradation could be significantly inhibited in the high dose PAE group (P < 0.01); the inhibition of PAE on IκBα degradation could be significantly inhibited in the SB group (P < 0.05). NF-κB/p65 signal was mainly located in cytoplasm in the normal group. NF-κB/p65 was translocated from cytoplasm to nucleus after stimulated by 45 min TNF-α in the TNF-α group, while it could be significantly inhibited in the high dose PAE group.

CONCLUSIONSPAE inhibited TNF-α-induced expression of lCAM-1. Its action might be associated with inhibiting TNFR1/NF-κB signaling pathway. p38 participated and mediated these actions.

Animals ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; Glucosides ; pharmacology ; Intercellular Adhesion Molecule-1 ; metabolism ; Mice ; Monoterpenes ; pharmacology ; NF-kappa B ; metabolism ; Receptors, Tumor Necrosis Factor ; metabolism ; Signal Transduction ; drug effects ; Tumor Necrosis Factor-alpha ; pharmacology

BACKGROUNDWe investigated the expression and role of TN4 in the oncogenesis of human hepatocellular carcinoma (HCC) from Qidong which is a HCC risk area.

METHODSThe expression of TN4 in HCC was observed using immunohistochemical staining (IHC). TN4 levels were manipulated in human liver cancer cell SMMC7721, using pcDNA3.1 eukaryotic expression constructs designed to express the complete TN4 cDNA. The biological changes of the cells were observed before and after transfection of TN4 and the change of gene expression was analysed by atlas cDNA expression array.

RESULTSAmong 100 pairs of samples of HCC, TN4 down-regulation expression and up-regulation expression positive rate were 81% (81/100), 19% (19/100), respectively (P < 0.01). TN4 protein was mainly localized in cytoplasm and membrane. The positive rate of TN4 were 10% (3/30), 100% (70/70) in lymph node metastasis and no lymph node metastasis, respectively (P < 0.01). The growth rates of the derivative SMMC7721-TN4 cell lines were decreased in comparison with that of normal SMMC7721 cells and pcDNA-SMMC7721. Some gene expression was changed before and after transfection of TN4. At 30 days of post-implantation of SMMC7721-TN4, SMMC7721-pcDNA3, SMMC7721 group produced tumors of (301.9 +/- 143.4) mm(3), (2418.7 +/- 362.8) mm(3), (2317.4 +/- 587.8) mm(3), respectively, (P < 0.01). Tumor inhibiting rate was 82.4% in TN4 transfection group. Sections of tumors were observed for their degree of tissue necrosis and there was higher degree of necrosis in tumors of the TN4-SMMC7721 cell group than those of the SMMC7721, SMMC7721-pcDNA groups.

CONCLUSIONSTN4 may play an important role in the oncogenesis of human HCC, especially in Qidong, the HCC risk area and TN4 could be a candidate tumor suppressor gene for HCC.

Adult ; Aged ; Carcinoma, Hepatocellular ; epidemiology ; genetics ; Carrier Proteins ; genetics ; China ; epidemiology ; DNA, Complementary ; analysis ; Gene Expression ; Genes, Suppressor ; Humans ; Immunohistochemistry ; Intracellular Signaling Peptides and Proteins ; Liver Neoplasms ; epidemiology ; genetics ; Lymphatic Metastasis ; genetics ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Myelin Proteins ; Nogo Proteins ; Risk Factors ; Transfection