The microstructure of cationic cyclopeptide (TD-34) treated Caco-2 cell membrane was observed, and we discussed the relationship between membrane structure and insulin transmembrane permeability. Atomic force microscope (AFM) was used to observe living cell membrane in air condition and tapping mode. Results showed that the surface of Caco-2 cell membrane treated with TD-34 lost its smoothness and nearly doubled its roughness. Apparent permeability coefficients (P(app)) of insulin in Caco-2 cell monolayers increased 2.5 times. In conclusion, AFM can be used to observe microstructure of cationic cyclopeptide treated cell membrane and cationic cyclopeptide enhanced insulin delivery across Caco-2 cell membrane by increasing membrane fluidity.
Caco-2 Cells ; Cations ; Cell Membrane ; drug effects ; Cell Membrane Permeability ; drug effects ; Humans ; Insulin ; metabolism ; Membrane Fluidity ; drug effects ; Microscopy, Atomic Force ; Peptides, Cyclic ; pharmacology

OBJECTIVE： To study the pharmacokinetics of domestic carvedilol and relative bioavailability of carvedilol capsule in Chinese volunteer.METHODS： Eight volunteers orally took a single dose of 30mg test preparation and 25mg control preparation in a random crossover and self-control method.Samples were determined by RP-HPLC fluorescent method.RESULTS： Profiles of carvedilol in vivo could be described as open two-compartment model.The main pharmacokinetics parameters of test and control preparations were as follows： Cmax（98.89± 27.60） ng/ml、 （70.06± 27.29） ng/ml， Tmax（0.4 849± 0.2 635） h、 （0.6 037± 0.1 707） h， CL（0.1 621± 0.08 057） （mg· h） /（ng· ml） 、 （0.1 796± 0.09 198） （mg· h） /（ng· ml） ， V/F(c)（0.2 127± 0.1 260） mg/(ng· ml)、 （0.2 777± 0.1 860） mg/（ng· ml） ， T1/2β （2.011± 1.709） h、 （1.959± 1.156） h， AUC（233.1± 97.12） ng/（ml· h） 、 （168.0± 70.61） ng/（ml· h） ； Mean relative bioavailability in man was (111.3± 15.18)％ .CONCLUSION： The results can be used for design of therapeutic scheme.

Adult ; Cranial Fossa, Middle ; anatomy & histology ; surgery ; Ear, Inner ; anatomy & histology ; surgery ; Facial Nerve ; anatomy & histology ; surgery ; Humans ; Male

Adenocarcinoma, Papillary ; blood supply ; metabolism ; pathology ; Adult ; Aged ; Carcinoma, Pancreatic Ductal ; blood supply ; metabolism ; pathology ; Chemokine CXCL12 ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lymph Nodes ; metabolism ; Lymphatic Metastasis ; Male ; Microvessels ; pathology ; Middle Aged ; Neoplasm Staging ; Neovascularization, Pathologic ; metabolism ; pathology ; Pancreas ; metabolism ; Pancreatic Neoplasms ; blood supply ; metabolism ; pathology ; Receptors, CXCR4 ; metabolism

Objective To investigate the clinical value of perforin and granzyme B expression in peripheral blood lymphocytes and diagnosis of acute rejection in renal transplantation. Methods Sixty-seven recipients of renal allograft were included in the study. The expression of perforin and granzyme B of peripheral blood lymphocytes was studied by quantitative reverse transcription polymerase chain reaction (RT-PCR). The recipients were divided into four groups, including 7 cases of acute rejection as group 1, 8 cases of delayed graft function as group 2, 27 cases of stable function as group 3, 25 cases of long-term survival as group 4. Results The expression of perforin and granzyme B in group 1 was significantly higher than that of the other three groups (P

Objective To investigate the effect of perforin and granzyme B in peripheral blood lymphocytes during acute rejection in renal transplantation.Methods Sixty-seven recipients of renal allograft were involved in the study.The recipients were divided into four groups: group 1 with 7 cases of acute rejection,group 2 with 8 cases of delayed graft function,group 3 with 27 cases of stable function and group 4 wih 25 cases of long-term survival.The expressions of perforin and granzyme B of peripheral blood lymphocytes were studied by quantitative reverse transcription polymerase chain reaction(RT-PCR).Results The expressions of perforin and granzyme B in acute rejection group were significantly higher than those of the other three groups(P

Objective To investigate the relationship between the preterm infants after premature rupture of the membranes(PROM)brain injury and some cellular factors in the umbilical cord blood and amniotic fluid,and ana-lyze the biological markers with great predictive value,and provide a theoretical basis for early monitoring of brain injury in premature infants. Methods One hundred and thirty - nine singleton infants with PROM,their gestation less than 34 weeks,were evaluated. The umbilical cord blood and amniotic fluid of cytokines,including interleukin - 1β(IL - 1β),IL - 4,IL - 6,IL - 8,IL - 10,IL - 17A,tumor necrosis factor alpha(TNF - α),granulocyte colony - stimu-lating factor(G - CSF),monocyte chemotactic protein - 1(MCP - 1),S100B protein and soluble intercellular adhe-sion molecule - 1(sICAM - 1)levels were measured with Luminex liquid chip. All the premature infants underwent brain imaging for the diagnosis of brain damage. All cases were divided into brain injury group and non - brain injury group based on brain imaging examination. Results The concentration of IL - 10 in cord blood was significantly lower in the brain injury group than that in the non - brain injury group,and the difference was statistically significant(P ﹤0. 05). The levels of IL - 1β,IL - 6,IL - 8,TNF - α,G - CSF,MCP - 1,S100B and sICAM - 1 in the brain injury group were significantly higher than those of non - brain injury group,and the differences were statistically significant (all P ﹤ 0. 05). The levels of IL - 1β,IL - 6,IL - 8,TNF - α,G - CSF,MCP - 1 and sICAM - 1 in the amniotic fluid were significantly higher than those of non - brain injury group,and the differences were statistically significant(all P ﹤ 0. 05),but amniotic fluid S100B protein level was similar between 2 groups,which had no statistical significance (P ﹥ 0. 05). To predict the value of brain damage in premature infants,the highest sensitivity in cord blood was S100B protein,the highest specificity was IL - 6. The highest sensitivity in amniotic fluid was IL - 1β,and the highest specificity was IL - 8. The levels of IL - 4 and IL - 17A in the umbilical cord blood and amniotic fluid,IL - 10 in amniotic fluid were very low,and had no predictive value for brain damage. Conclusions Many biological markers in umbilical cord blood and amniotic fluid provide information about the risk of brain injury in premature infants. The highest sensitivity in cord blood was S100B protein,the highest specificity was IL - 6. The highest sensitivity in amniotic fluid was IL - 1β,the highest specificity was IL - 8. Changes in inflammation - related biomarkers suggest that brain damage in the preterm infants might be associated with intrauterine inflammation.

This study is to investigate the effect of artesunate on transforming growth factor-beta1 (TGF-beta1) induced epithelial-mesenchymal transition (EMT) and its possible mechanism. After the in vitro cultured RLE-6TN cells were treated with TGF-beta1 then artesunate intervened on it, after 24 h, expression of the markers of mesenchymal cell was assayed using Western blotting and real-time PCR analysis. Western blotting was also used to detect the effect of TGF-beta1 on the Smad3 and Smad7 expressions of RLE-6TN cells. Morphological alterations were examined by phase-contrast microscope, and ultrastructure changes by electron microscope. Incubation of RLE-6TN cells with TGF-beta1 resulted in the up-regulation of the expression of the mesenchymal cell markers, after artesunate intervened on it, resulted in the down-regulation of the expression. Meanwhile, incubation with artesunate intervened on RLE-6TN cells could lead to the apparent down-regulation of the expression of Smad3 and up-regulation of Samd7 and the transition of RLE-6TN cells to mesenchymal-like by TGF-beta1 induction, after artesunate intervened on it, RLE-6TN cells to epithelial-like. TGF-beta1 induced epithelial-mesenchymal transition process; artesunate can inhibit TGF-beta1-induced epithelial-mesenchymal transition process, the possible mechanism is up-regulation of the expression of Smad7 and down-regulation of the expression of Smad3, meanwhile inhibits phosphorylation of Smad3.
Actins ; genetics ; metabolism ; Animals ; Artemisia ; chemistry ; Artemisinins ; isolation & purification ; pharmacology ; Cell Line ; Cell Proliferation ; drug effects ; Epithelial Cells ; cytology ; metabolism ; Epithelial-Mesenchymal Transition ; drug effects ; Idiopathic Pulmonary Fibrosis ; pathology ; Plants, Medicinal ; chemistry ; Pulmonary Alveoli ; cytology ; RNA, Messenger ; metabolism ; Rats ; Smad3 Protein ; genetics ; metabolism ; Smad7 Protein ; genetics ; metabolism ; Transforming Growth Factor beta1 ; pharmacology ; Vimentin ; genetics ; metabolism

Objective:To investigate the characteristics and significance of insulin-like growth factor 1(IGF- 1) and insulin-like growth factor binding protein 2 (IGFBP2) expressions in ankle cartilage of patients with Kashin-Beck disease (KBD).Methods:In this case-control study, 10 KBD patients who were hospitalized in the Department of Orthopedics of Shaanxi Provincial People's Hospital from January 2010 to December 2016 were selected as KBD group, and 10 patients with ankle fracture caused by trauma but without talus injury during the same period were selected as control group, the cartilage tissues of the two groups were collected. IGF-1, IGFBP2 positive cells, the mRNA and protein expressions of IGF-1, IGFBP2 in the cartilage tissues were detected by immunohistochemistry, real-time fluorescent quantitative PCR and Western blotting, respectively. According to the expressions of IGF-1 and IGFBP2 in ankle cartilage of KBD patients, a patient with amputation caused by trauma was selected in Shaanxi Provincial People's Hospital, and ankle joint cartilage was taken to prepare chondrocytes for in vitro cell verification experiments. The chondrocyte were divided into control group (0 ng/ml T-2 toxin), T-2 treatment group (20 ng/ml T-2 toxin) and T-2+ IGFBP2 silenced group (20 ng/ml T-2 toxin+ 50 nmol/L IGFBP2 siRNA), the MTT method and dimethyl methylene blue staining were used to detect the activity of chondrocyte and the secretion of sulfated glycosaminoglycan (sGAG). Results:In the control group and the KBD group, the number of IGF-1[(47.26 ± 8.97), (68.15 ± 7.42) cells] and IGFBP2 positive cells [(27.56 ± 5.40), (71.85 ± 7.62) cells] in the cartilage tissues were significantly different ( t = 4.487, 9.402, P < 0.01). Compared with the control group, the IGF-1, IGFBP2 mRNA and protein expression levels in KBD group were significantly higher, the differences were significantly different ( t = 3.340, 20.700, 4.684, 8.699, P < 0.05 or < 0.01). In cell experiment, the chondrocyte activitives and sGAG contents of the control group, T-2 treatment group, and T-2+ IGFBP2 silenced group were significantly different ( F = 226.70, 80.66, P < 0.01); among them, the cell activitives and sGAG contents of the T-2 treatment group and T-2+ IGFBP2 silenced group were lower than those of control group ( P < 0.05), and the T-2+ IGFBP2 silenced group were higher than those of the T-2 treatment group ( P < 0.05). Conclusions:The expressions of IGF-1 and IGFBP2 in the ankle cartilage of KBD patients are significantly higher. Silencing IGFBP2 gene can reduce the inhibitory effect of T-2 toxin on chondrocyte activity and the secretion of sGAG.

OBJECTIVETo investigate the effects of butyl alcohol extract of baitouweng decoction (BAEB) on the fungal cell surface hydrophobicity (CSH), filamentation and biofilm formation of Candida tropicalis.

METHODGradual dilution method was used to determine the MIC. XTT assay was applied to determine the SMIC80. Time-Kill assay was employed to draw the Time-Kill curve. The water-hydrocarbon two-phase assay was used to measure the cell surface hydrophobicity. Scanning electron microscopy (SEM) was applied to observe the morphological changes of the biofilm. Confocal laser scanning microscopy (CLSM) was applied to determine the thickness of the biofilm. The quantification real-time PCR (qRT-PCR) was used to detect expression changes of releated genes (UME6, ALST3 and NRG1). result: The MICs of BAEB against C. tropicalis strains are determined as 64-128 mg x L(-1). The SMIC80 s of BAEB against the biofilm of Candida tropicalis strains are determined as 256-512 mg x L(-1). Time-Kill curve results indicate that BAEB has a promise fungicidal effect at 256 and 512 mg x L(-1). SEM results shows that 512 mg x L(-1) BAEB can inhibit the formation of C. tropicalis biofilm on Silicone catheter, and the morphology of biofilm is also affected by BAEB. The thickness of C. tropicalis biofilm is reduced by BAEB according to CLSM results. Furthermore, qRT-PCR results indicate that expression of UME6 and ALST3 are significantly down-regulated by BAEB 256,512 mg x L(-1), and NRG1 is not affected by BAEB.

CONCLUSIONBAEB inhibits effectively the CSH, filamentation and biofilm formation of VVC strains of C. tropicalis.

Antifungal Agents ; chemistry ; pharmacology ; Biofilms ; drug effects ; Candida tropicalis ; drug effects ; genetics ; physiology ; Candidiasis ; microbiology ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Fungal Proteins ; genetics ; metabolism ; Gene Expression Regulation, Fungal ; drug effects ; Humans ; Virulence Factors ; genetics ; metabolism