Objective The aim of this study was to explore possible pharmacokinetic factors and develop a population pharmacokinetic model for remifentanil in adult patients.Methods Eleven healthy patients,undergoing elective major abdominal surgery,aged 25 to 86 years,received random-ly remifentanil 0.3μg·kg-1 ·min-1 (group R3),or 0.6μg·kg-1 ·min-1 (group R6).Frequent ar-terial blood samples were drawn according to predetermined time and assayed for remifentanil concen-tration.Nonlinear mixed-effects modeling (NONMEM)was used to evaluate the time courses of the measured concentrations.The covariates include age,bodyweight (WT),gender,lean body mass (LBM),body mass index (BMI)and body surface area (BSA).Results The pharmacokinetic data of remifentanil were well described using a three-compartment linear model with first-order elimination from the central compartment.Forward analysis showed that age,height and body mass index (BMI) does not affect the pharmacokinetic parameters,which are contrast with body weight,lean body mass (LBM),body surface area (BSA)and gender;further analysis demonstrated only a significant effect of body weight on remifentanil systemic clearance (CL)and volume of the central compartment (V). For typical 60 years patients,PK parameters were:V1 =7.64 L,V2 =4.81 L,V3 =4.34 L,CL1 =2.74 L/min,CL2 = 0.738 L/min,CL3 = 0.0905 L/min.Conclusion The pharmacokinetics of remifentanil is consistent with its rapid elimination by blood and tissue esterase in Chinese patients. The systemic clearance and volume of distribution of central compartment increases with body weight in the population and the range of covariates studied,which suggests that a patient with greater body weight needs a greater initial dose and maintenance infusion rate higher to obtain a stable plasma con-centrations and clinical effects.

Objective:To observe the expression of microRNA (miRNA, miR) -7850 in renal cancer tissues, and to explore the effect of miR-7850 on the proliferation and migration of renal cancer cells and on the regulation of serine proteinase inhibitor B3 (SERPINB3) gene expression.Methods:Real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-7850 in renal cancer tissues and renal cancer cell lines. The renal cell carcinoma cell line with the lowest expression of miR-7850 was selected, and the negative control sequence (miR-NC) and miR-7850 mimics were transfected into renal cell carcinoma cells by Lipofectamine 2000 transfection reagent, respectively, which were defined as miR-NC group and miR-7850 group. qRT-PCR was used to detect the expression of miR-7850 in transfected renal cancer cells. The cell proliferation and migration ability after transfection were detected by cell counting kit-8 (CCK-8) method and transwell experiment. Bioinformatics prediction and dual luciferase reporter gene experiments were used to verify the target gene of miR-7850. qRT-PCR and Western blot were used to detect the expression of target genes in renal cancer cells after transfection.Results:Compared with adjacent tissues (5.95±0.44), the expression of miR-7850 in kidney cancer tissues (1.19±0.33) was lower ( P<0.01). Compared with immortalized proximal renal tubular epithelial cells (1.01±0.07), the expression of miR-7850 was lower in renal cancer cell lines ( P<0.05), and the lowest in A498 cells (0.13±0.01) ( P<0.01). The expression of miR-7850 in the miR-7850 group (7.46±0.93) was significantly higher than that in the miR-NC group (1.01±0.08) ( P<0.01), indicating successful transfection. Compared with the miR-NC group, the cell proliferation ability of the miR-7850 group was significantly reduced ( P<0.05). The number of migrating cells in miR-NC group and miR-7850 group were (139.50±12.31) and (75.09±16.05) cells, respectively, and the cell migration ability in miR-7850 group decreased significantly ( P<0.01). Bioinformatics technology shows that the target gene of miR-7850 was SERPINB3. The dual luciferase reporter gene experiment confirmed that miR-7850 can target the SERPINB3 gene ( P<0.05). Compared with the miR-NC group, the expression of SERPINB3 in cells of miR-7850 group was significantly reduced ( P<0.05), as well as the CDK4, CyclinD, Snail and Vimentin. Conclusions:miR-7850 is lowly expressed in renal cancer tissues and cell lines. miR-7850 can inhibit the proliferation and migration of renal cancer A498 cells, which may be related to its inhibition of SERPINB3 gene expression.

AIM:To compare the higher-order aberrations between implantations of AcrySof and AcrySof ReStor multifocal aspherical intraocular lens(IOL) with various pupil diameters.METHODS:Fifty-four patients(62 eyes)with bilateral senile cataracts were retrospectively selected.Patients were operated with phacoemulsification and IOL implantation.They were divided into two groups based on the IOL implantation of AcrySof IQ and AcrySof ReStor.Wave front aberration:spherical aberration(C12)and the root mean square of the total higher-order aberration(RMSh)were observed 3 months after surgery.RESULTS:The larger the pupil was, the higher the C12 and RMSh were in the eyes (P<0.01).There were no statistical differences in C12 or RMSh between two groups at 5, 6 or 7mm pupil diameters (P>0.05). CONCLUSION:There are no differences between AcrySof IQ group and AcrySof ReStor group at 5, 6 or 7mm pupil diameters.

Objective To explore the relationship between the apoptosis of esophageal carcinoma tissue and its adjacent tissues with the survival time of patients with esophageal carcinoma by examining the apoptosis in esophageal carcinoma tissue and its adjacent tissues.Methods FCM Wag performed to detect the rates of apoptosis in 68 cases of esophageal carcinoma and their adjacent tissues and 28 cases of normal esophageal mucosae tissue.Sixty-three patients with esophageal carcinoma had been followed up and noted the survival time of every patient from the operated day to the deadline.Results The rate of apoptosis was the highest in the normal esophageal mucosae tissue(12.78±1.32)%,(7.79±1.48)% in adjacent tissue,and(4.16±2.06)% in esophageal carcinoma tissue,respectively.To follow the survival time after operation of 63 cases with esophageal carcinoma showed 60 cases in one year survival time and 35 cases in three years survival time.The rate of apoptosis in the esophageal carcinoma and its adjacent tissues wag(3.45±1.51)% and (3.96±0.94)% in the patients of one year survival time,(3.90±2.53)% and (7.89±2.27)% in the patients of three years survival time,respectively,P<0.05.Conclusions There is the phenomenon of apoptosis-escape in the esophageal carcinogenesis.There is close relationship between the apoptosis of esophageal carcinoma tissue and the survival time after operation.The apoptosis in the adjacent tissue is more important than that in the esophageal carcinoma tissue for evaluating the survival time after operation of patients with esophageal carcinoma.

Objective To study the expression of early growth response gene-1(Egr-1)in esophageal carcinoma tissue,and to explore the relationship between Egr-1 and the survival time of the patients with esophageal carcinoma.Methods RT-PCR was performed to detect the expression of Egr-1 mRNA in68 cases of esophageal carcinoma.The relationship between survival time and prognosis was analyzed.Results The expression of Egr-1 mRNA was the lowest(0.567±0.404),(0.945±0.336)and(1.201±0.347)in esophageal carcinoma tissue,para-cancerous tissue and normal esophageal mucosa tissue,respectively(F=12.709,P＜0.05,P＜0.00).21 cases showed the positive expression of Egr-1 mRNA of both the esophageal carcinoma tissue and the para-cancerous tissue.21 cases showed the positive expression of Egr-1 mRNA in a single esophageal carcinoma tissue or the para-cancerous tissue.26 cases showed the negative expression of Egr-1 mRNA both in the esophageal carcinoma tissue and the paracancerous tissue.The positive rate of Egr-1mRNA expression was 65.71%and 30.00%in the groups of the survival time for three years and the groups of the survival time for one year(P＜0.05).The survival rates in the two groups with positive expression of Egr-1 mRNA were 94.44%and 86.96%,respectively(P＞0.05).Conclusion Decreased level of EGR1 expression may be related to esophageal carcinogenesis.The expression level of Egr-1 mRNA might be associated with the survival time of the patients with esophageal carcinoma.Egr-1 expression in esophageal carcinoma tissue may be of great value for determining prognosis of esophageal carcinoma.

The aim of this study was to isolate, cultivate and phenotypically characterize two types of human amnio-tic membrane (HAM)-derived cells, and to analyze their differentiation potential in vitro. Human amnion epithelial cells (hAEC) were derived from the embryonic ectoderm, while human amnion mesenchymal cells (hAMC) were derived from the embryonic mesoderm. The cells were characterized by flow cytometry and immunofluorescence, then immunofluorescence also was performed for the analysis of multipotentiality in differentiation. The results indicated that immunophenotypic characterization of both cell types demonstrated positive for HLA-A, B, C and mesenchymal stem cell markers (CD29, CD73, CD44, CD59, CD90, CD105, CD166), but did not express the hematopoietic markers (CD31, CD34, CD45, HLA-DR) and showed the weak expression of costimulatory molecules (CD40, CD40L, CD80, CD86). Phenotypes of both cell populations were maintained from passages 3 to 7. The immunofluorescence indicated that hAEC expressed cytokeratin 19, but did not express vimentin. On the contrary, hAMC expressed vimentin but did not express cytokeratin 19. The assessment of multilineage potential demonstrated that hAMC showed greater cardiomyocytes potential, while hAEC showed greater neural potential. It is concluded that hAEC and hAMC can be successfully isolated from the HAM. Both cell populations possess similar immunophenotype. However, they differ in cell yield and multipotential for differentiation into the major lineages, hAEC possess a much greater ectodermal differentiation capacity, while hAMC possess a much greater mesodermal differentiation capacity. This conclusion will be important for use of these cells in cell therapy.
Amnion ; cytology ; Cell Differentiation ; physiology ; Cell Lineage ; Epithelial Cells ; cytology ; Humans ; Immunophenotyping ; Stromal Cells ; cytology

Human leukocyte antigen G (HLA-G), a kind of non-classical major histocompatibility complex class I antigens, can inhibit inflammatory reaction, assist tumor cells to escape from immune surveillance and promote the immunologic tolerance of the graft. HLA-G, expressed and secreted by human amniotic mesenchymal cells (HAMC), suppresses the functions of NK cells, T cells and B cells and modulates the activity of dendritic cells (DC). These findings provide a theoretical basis for illustrating the mechanism of immunosuppression on HAMC. In this article, the recent advances on not only the gene and the molecular structure of HLA-G, but also the possible mechanisms of HLA-G in immunoregulatory function of HAMC, as well as the relation of HLA-G with HAMC, NK, DC, T and B cells are reviewed.
Amnion ; cytology ; HLA-G Antigens ; immunology ; Humans ; Immune Tolerance ; Mesenchymal Stromal Cells ; cytology ; immunology

The present study is to establish Caco-2/HT29-MTX co-cultured cells and investigate the transport capability of PLGA nanoparticles with different surface chemical properties across Caco-2/HT29-MTX co-cultured cells. PLGA-NPs, mPEG-PLGA-NPs and chitosan coated PLGA-NPs were prepared by nanoprecipitation method using poly(lactic-co-glycolic acid) as carrier material with surface modified by methoxy poly(ethylene glycol) and chitosan. The particle size and zeta potential of nanoparticles were measured by dynamic light scattering. Coumarin 6 was used as a fluorescent marker in the transport of nanoparticles investigated by confocal laser scanning microscopy. The transport of furanodiene (FDE) loaded nanoparticles was quantitively determined by high performance liquid chromatography. Colchicine and nocodazole were used in the transport study to explore the involved endocytosis mechanisms of nanoparticles. Distribution of the tight junction proteins ZO-1 was also analyzed by immunofluorescence staining. The results showed that the nanoparticles dispersed uniformly. The zeta potential of PLGA-NPs was negative, the mPEG-PLGA-NPs was close to neutral and the CS-PLGA-NPs was positive. The entrapment efficiency of FDE in all nanoparticles was higher than 75%. The transport capability of mPEG-PLGA-NPs across Caco-2/HT29-MTX co-cultured cells was higher than that of PLGA-NPs and CS-PLGA-NPs. Colchicine and nocodazole could significantly decrease the transport amount of nanoparticles. mPEG-PLGA-NPs could obviously reduce the distribution of ZO-1 protein than PLGA-NPs and CS-PLGA-NPs. The transport mechanism of PLGA-NPs and mPEG-PLGA-NPs were indicated to be a combination of endocytosis and paracellular way, while CS-PLGA-NPs mainly relied on the endocytosis way. PEG coating could shield the surface charge and enhance the hydrophilicity of PLGA nanoparticles, which leads mPEG-PLGA-NPs to possess higher anti-adhesion activity. As a result, mPEG-PLGA-NPs could penetrate the mucus layer rapidly and transport across Caco-2/HT29-MTX co-cultured cells.
Biological Transport ; Caco-2 Cells ; Chitosan ; chemistry ; Coated Materials, Biocompatible ; chemistry ; Coculture Techniques ; Drug Carriers ; Furans ; administration & dosage ; chemistry ; metabolism ; HT29 Cells ; Heterocyclic Compounds, 2-Ring ; administration & dosage ; chemistry ; metabolism ; Humans ; Lactic Acid ; chemistry ; Nanoparticles ; Particle Size ; Polyethylene Glycols ; chemistry ; Polyglycolic Acid ; chemistry ; Zonula Occludens-1 Protein ; metabolism

Objective To identify seven species of Gynostemma BI.,including G.pentaphyllum,G. pentagynum,G.cardiospermum,G.longipes,G.yixingense,G.laxiflorum,and G.guangxiense,by in- ter-simple sequence repeat(ISSR)markers.Methods General DNA was isolated from leaves of the seven species in Gynostemma B1.by CTAB,57 primers constituted by ISSR were tested for PCR and sepharose electrophoresis.Results Fourteen primers amplified polymorphic bands,the amplification patterns of primers UBC-873 and UBC-895 were higher in terms of polymorphic and amplified band ratio.They are used to distinguish all the examined seven species.Conclusion ISSR-PCR Method provides a quick,reli- able molecular marker technique for the identification of different species of Gynostemma B1.

Objective:To explore the expression of long non-coding RNA LINC00630 in bladder cancer cell lines, and to explore the effect of interference with its expression in vitro on the proliferation and migration of bladder cancer cells.Methods:Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of LINC00630 in bladder cancer cell lines 5637, BIU-87, T24, J82 and normal bladder epithelial cell line SV-HUC-1. The bladder cancer cell line with the highest LINC00630 expression was selected for follow-up experiments, then the cell line infected with the control lentivirus was used as the control group, and the cell line infected with the lentivirus that could interfere with the expression of LINC00630 was used as the experimental group. qRT-PCR was used to detect the expression of LINC00630 in the two groups of cells. MTS method and cell scratch test were used to detect the proliferation and migration abilities of cells in the two groups. qRT-PCR was used to detect the expression of neuregulin 1 (NRG1) mRNA in the two groups of cells, and Western blot was used to detect the expressions of NRG1 protein, cell proliferation-related proteins (cyclin D3 and CDK2) and cell migration-related proteins (Vimentin and N-cadherin) in the two groups of cells.Results:Compared with SV-HUC-1 cells (1.05±0.17), the expression of LINC00630 was significantly increased in all bladder cancer cell lines (all P < 0.01), and the expression was highest in J82 cells (relative expression 5.83±0.42). Compared with J82 cells of the control group, the expression of LINC00630 in J82 cells of the experimental group decreased (0.18±0.02 vs. 1.00±0.05, t=14.36, P < 0.01); from day 2 of transfection, the cell proliferation activity of the experimental group was lower than that of the control group (all P < 0.05). The cell scratch closure rate of the experimental group was lower than that of the control group [(27.4±7.1)% vs. (66.0±5.4)%, t = 4.31, P < 0.01]. Therelative expression of NRG1 mRNA in the experimental group was lower than that in the control group (0.34±0.03 vs. 1.07±0.24, t = 2.99, P < 0.05). Compared with the control group, the expressions of NRG1 protein, cell proliferation-related proteins and cell migration-related proteins in the experimental group were reduced. Conclusions:LINC00630 is up-regulated in bladder cancer cell lines, and interference with LINC00630 may inhibit the proliferation and migration of J82 cells by down-regulating the expression of NRG1 gene. LINC00630 may be a new molecular target for the treatment of bladder cancer.