Acupuncture Analgesia ; Acupuncture, Ear ; Adult ; Chemoembolization, Therapeutic ; Humans ; Liver Neoplasms ; complications ; surgery ; Male ; Pain, Postoperative ; etiology ; therapy

The aim of this paper was to investigate the effect of terpene penetration enhancers on membrane fluidity and membrane potential using HaCaT keratinocytes, and study the potential mechanisms of these terpene compounds using as natural transdermal penetration enhancer. Six terpene compounds, namely menthol, limonene, 1,8-cineole, menthone, terpinen-4-ol and pulegone, were chosen in this study on account of their good penetration-enhancement activities. The cytotoxicity of these terpene compounds was measured using an MTT assay. The fluorescence recovery after photobleaching (FRAP) technique was employed to measure the change of membrane fluidity of HaCaT cells. The flow cytometer was used to study the alteration of membrane fluidity of HaCaT cells, and investigate the effect of terpene compounds on intracellular Ca2+. It was found that 6 terpene compounds possessed low cytotoxicity in comparison to the well-established and standard penetration enhancer azone. Those terpene compounds could significantly enhance HaCaT cells membrane fluidity and decrease HaCaT cells membrane potentials. Meanwhile, after treated with various terpene compounds, the Ca2(+)-ATPase activity and intracellular Ca2+ of HaCaT cells was decreased significantly. Terpene penetration enhancers perhaps changed the membrane fluidity and potentials of HaCaT cells by altering the Ca2+ balance of the cell inside and outside, resulting in the low skin permeability to increase the drug transdermal absorption.
Cell Line ; Drugs, Chinese Herbal ; pharmacokinetics ; Humans ; Keratinocytes ; drug effects ; metabolism ; Membrane Fluidity ; drug effects ; Skin Absorption ; drug effects ; Terpenes ; pharmacokinetics

OBJECTIVETo observe the effect of umbilical sticking therapy (UST) with Qitou Xiaugu Plaster (QXP) on hemodynamics of portal system in patients with liver cirrhosis.

METHODSOne hundred and twenty patients of liver cirrhosis with portal hypertension were assigned to two groups. On the basis of conventional therapy, UST was applied in the 66 patients in treated group, which was exchanged once every 3 days with an interval of 1-day rest. The 54 patients in the control group were orally administered with propanolol. The therapeutic course for both groups was 1 month. Before and after treatment, the hemodynamic changes in portal or splenic veins were observed by color Doppler ultrasonograph, and the changes of liver function, blood coagulation and patients' subjective symptoms were observed as well.

RESULTSAfter treatment, portal vein diameter and splenic vein diameter significantly decreased (P < 0.05, portal venous flow velocity and splenic venous flow velocity apparently increased (P < 0.05), and portal venous flow apparently decreased in both groups (P < 0.05), while no significant change was found in the splenic venous flow (P > 0.05). The liver function and blood coagulation indexes in both groups were improved. The improvement of clinical symptoms in the treated group was superior to that in the control group.

CONCLUSIONUST with QXP could decrease the portal vein pressure in a short time, with the therapeutic effect comparable to propanolol, and with no adverse reaction.

Adult ; Aged ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hemodynamics ; drug effects ; Humans ; Hypertension, Portal ; drug therapy ; physiopathology ; Liver Cirrhosis ; drug therapy ; physiopathology ; Male ; Middle Aged ; Portal Pressure ; drug effects ; Portal Vein ; drug effects ; physiopathology ; Splenic Vein ; drug effects ; physiopathology ; Umbilicus ; blood supply ; Young Adult

Background Studies demonstrated that human amniotic epithelial cells (AECs) have some characteristics of embryonic stem cells and they were used to re-establish the surface of eyes. Human AECs may serve as new seed cells in tissue engineering for corneal epithelium reconstitution in the future. Objective The present study was to investigate the application value of human amniotic epithelium cells transfected by lentiviral vectormediated enhanced green fluorescent protein (EGFP) gene as new seed cell source for engineering the corneal surfacelayer. Methods Lentiviral vector carrying the objective gene EGFP was transfected into human amniotic epithelial cells (pLenti6/V5-DEST),and the transient expression of the transgene in the human amniotic epithelial cells was observed under the fluorescence microscope. Flow cytometry was used to detect the positive expression rates of EGFP in transfected cells. The transfected human amniotic epithelial cells were seeded onto the fresh corneal stromal surface of New Zealand white rabbit and cultured in vitro. The stem cell deficiency ( SCD ) models were established by cutting off the limbus of cornea in 20 eyes of New Zealand white rabbits, and the model rabbits were then divided into 2 groups randomly. The transplanted grafts carrying the pLenti6/V5-DEST-EGFP gene-transferred human amniotic epithelium cells were regarded as the pLenti6/V5-DEST-EGFP group, and the corneal stroma graft without any epithelial cell served as the control group. The opacity of stroma and corneal conjunctivalization and vascularization were observed daily. The rabbits' eyes were extracted one month after operation. The expression of EGFP in the cornea was detected under the fluorescence microscope, and the expression of CK8, CK18 and CK12 in cornea was detected by immunohistochemical staining. Results The shape of the transferred human amniotic epithelial cells resembled normal human amniotic epithelial cells. 48 hours after the transient transfection of EGFP presented with the highest expression level throughout the observation duration, with a positive expression rate of EGFP of 61.5% ,showing significant differences in comparison with that of 12 ( 5.24% ) , 24 ( 38.27% ) or 96 ( 39. 10% ) hours ( P ＜0. 05) post-transfection; but no obvious difference was found in the positive rate of transiently transfected EGFP between 48 hours and 72 hours ( 58.36% ) ( P＞0. 05 ). Six cornea grafts were clear in 1 month and two corneas were rejected during the observation period in the pLenti6/V5-DEST-EGFP group. A few new blood vessels were seen around the graft. Ten corneas of the control group became opaque and cloudy with new blood vessels growth around the grafts. Imunohistochemistry revealed the positive expressions of CK8, CK1 8 and CK12 in the corneal epithelial layer in the pLenti6/V5-DEST-EGFP group. However,the expression of CK12 was absent in the control group. Conclusion Human amniotic epithelium cells transfected with the pLenti6/V5-DEST-EGFP gene is a new and ideal feed cell type to reconstruct the corneal surface layer. Lentivirus is a relatively safe gene transfection vector.

Objective To explore the treatment of non-remission of trigeminal neuralgia after microvascular decompression (MVD). Methods The data of 50 patients with trigeminal neuralgia from 2005 to 2009 were reviewed and analyzed; those with non-remission of trigeminal neuralgia after MVD were performed percutaneous shutdown of trigeminal cistern with glycerol via foramen ovale. The operative efficacy was compared. Results Three (6%) patients were noted with non-remission outcomes: 2 with blood vessel compression were performed MVD and 1 without blood vessels compression did not receive MVD procedure; all these 3 patients had thicken adhesion of the arachnoid.The pain was remitted after the showdown operation. Conclusion Percutaneous shutdown oftrigeminal cistern with glycerol is a safe and effective surgical method for patients with non-remission oftrigeminal neuralgia after MVD.

Objective To investigate the influence of RNA-interfered ribonucleotide reductase M2(RRM2)on apoptosis and invasion of ovarian cancer cells with drug resistant.Methods The specific small interfering(siRNA)transfected SKOV3/DDP of RRM2 gene were designed as the interference group.SKOV3/DDP cells,SKOV3/DDP-RRM2 non-specific negative cells were designed as the blank group and negative group.The cell proliferation inhibition rate was detected,RI and drug resistant transfection rate were calculated,the expression of RRM2 mRNA was detected by fluorescence PCR.The effect of siRNA transfection on SKOV3/DDP resistance index and RRM2 protein was analyzed,and the invasion ability of SKOV3/DDP cells was observed with Transwell.Results The transfection rate of the blank group,negative group and interference group was 90％.The drug resistance of DDPto SKOV3/DDP cells was low and gemcitabine was sensitive to SKOV3/DDP cells.DDP and gemcitabine of the DDP on cell half inhibitory concentration(IC50)values showed significant differences among three groups(P<0.05).IC50 values of DDP and gemcitabine on cells in the interference group were significantly lower than the negative group and blank group(P<0.05).The relative expression of SKOV3/DDP cells and SKOV3 cells RRM2 protein showed significantdifferences(P<0.05),and the relative expression was significantly lower in transfection group I,transfection group II and transfection group III than the negative group and blank group(P<0.05).The decrease of relative expression of RRM2 protein was the greatest in the transfection group I(P<0.05).The apoptosis rate was significantly higher in the interference group than the blank group and the negative group,transmembrane cells were fewer than the blank group and the negative group(P<0.05).Conclusion siRNA can effectively inhibit the proliferation and invasion of RRM2 gene in ovarian cancer.It can increase the drug sensitivity of drug resistant cells,especially cells promoting the apoptosis of drug resistant cells induced by DDP.

Objective To investigate the influence of RNA-interfered ribonucleotide reductase M2(RRM2)on apoptosis and invasion of ovarian cancer cells with drug resistant.Methods The specific small interfering(siRNA)transfected SKOV3/DDP of RRM2 gene were designed as the interference group.SKOV3/DDP cells,SKOV3/DDP-RRM2 non-specific negative cells were designed as the blank group and negative group.The cell proliferation inhibition rate was detected,RI and drug resistant transfection rate were calculated,the expression of RRM2 mRNA was detected by fluorescence PCR.The effect of siRNA transfection on SKOV3/DDP resistance index and RRM2 protein was analyzed,and the invasion ability of SKOV3/DDP cells was observed with Transwell.Results The transfection rate of the blank group,negative group and interference group was 90％.The drug resistance of DDPto SKOV3/DDP cells was low and gemcitabine was sensitive to SKOV3/DDP cells.DDP and gemcitabine of the DDP on cell half inhibitory concentration(IC50)values showed significant differences among three groups(P<0.05).IC50 values of DDP and gemcitabine on cells in the interference group were significantly lower than the negative group and blank group(P<0.05).The relative expression of SKOV3/DDP cells and SKOV3 cells RRM2 protein showed significantdifferences(P<0.05),and the relative expression was significantly lower in transfection group I,transfection group II and transfection group III than the negative group and blank group(P<0.05).The decrease of relative expression of RRM2 protein was the greatest in the transfection group I(P<0.05).The apoptosis rate was significantly higher in the interference group than the blank group and the negative group,transmembrane cells were fewer than the blank group and the negative group(P<0.05).Conclusion siRNA can effectively inhibit the proliferation and invasion of RRM2 gene in ovarian cancer.It can increase the drug sensitivity of drug resistant cells,especially cells promoting the apoptosis of drug resistant cells induced by DDP.

Abstract?AIM: To investigate mechanism of bradykinin ( BK) on inflammations of retinal pigment epithelium ( RPE) cells.?METHODS: ARPE -19 cells were cultured in vitro, stimulated by 100nM BK for 24h. Cell morphology changes were observed by microscope, and BK receptor localization was detected through cell immunofluorescence. Changes of Ca2+in BK and BR antagonist stimuli were detected by laser scanning confocal microscopy.The expressions of COX-1, COX-2, eNOS and iNOS protein in control group and BK group were detected by Western Blot.?RESULTS: After the stimulation of BK, there was no significant changes of ARPE-19 cells in morphology.Kinin B1 receptors ( B1R ) and B2 receptors ( B2R ) could be detected in ARPE-19 cells.Compared with control group, Ca2+concentrations significantly increased in BK group; in B1R antagonist group and B2R antagonist group Ca2+concentrations increased less than BK group; B1R and B2R antagonist group showed no obvious changes in Ca2+concentrations.Compared with control group, COX-2 and iNOS protein concentrations were significantly increased in BK group (P<0.001).?CONCLUSION:BK induces the increasing expression of COX-2 and iNOS in the cultured ARPE cells through binding with either B1R or B2R.

OBJECTIVETo observe the effect of Xuefu Zhuyu decoction (XZD) on the chronic hepatitis B caused liver fibrosis (CHBLF) and the portal hemodynamics.

METHODSSixty patients with CHBLF were randomly divided into two groups, the 28 patients in the treated group were treated with oral intake of XZD and conventional liver protective treatment, the 32 patients in the control group were treated with conventional liver protective treatment alone, the therapeutic course for both groups was 3 months. Serum liver fibrosis criteria and portal dynamical criteria were observed before and after treatment.

RESULTSComparison of the remarkable effective rate between the two groups showed significant difference. After treatment, in the treated group, all the serum criteria for liver fibrosis (HA, PCIII, LN) and criteria for portal trunk hemodynamics, such as mean velocity and quantity of blood flow were significantly improved (P < 0.05 or P < 0.01), as compared with those in the control group, the difference was also significant (P < 0.05 or P < 0.01).

CONCLUSIONXZD has definite therapeutic effect on chronic hepatitis B caused liver fibrosis.

Adolescent ; Adult ; Collagen Type III ; blood ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hemodynamics ; Hepatitis B, Chronic ; blood ; complications ; Humans ; Hyaluronic Acid ; blood ; Liver Cirrhosis ; blood ; drug therapy ; etiology ; Male ; Middle Aged ; Phytotherapy ; Portal Vein ; physiopathology

OBJECTIVETo construct a recombinant adenovirus vector that expresses small hairpin RNAs (shRNA) against COX-2, AKT1 and PIK3R1 gene and to evaluate its potential for suppressing the cell proliferation of human gastric adenocarcinoma SGC701 cell in vitro and in vivo, which will enable the development of a gene therapy protocol for the treatment of human gastric adenocarcinoma.

METHODSThree strips of shRNA targeting AKT1, COX-2 and PIK3R1, was subcloned into adenovirus expression vector. After verification, it was amplified and titered. The recombinant adenovirus expression vector was infected into human gastric adenocarcinoma SGC7901 cells in vitro and the infected cells were injected in nude mice. The mRNA and protein expression levels of AKT1, COX-2 and PIK3R1 were determined by real-time PCR and Western blot respectively. Cell proliferation in vitro was determined by methyl thiazolyltetrazolium (MTT) assay and flow cytometry, tumor growth in vivo was measured by volume of tumor in nude mice.

RESULTSRestriction digestion and sequencing analysis showed that the rAd5-C-A-P adenovirus expression vector was constructed successfully. It significantly inhibited the expression of AKT1, COX-2 and PIK3R1, and cell growth was inhibited over 70% as indicated by MTT assay and accompanied with G0/G1 phase arrest. Cell growth on matrigel matrix showed that the rAd5-C-A-P transfected cells were detached from the matrix or grew in a scattered clustering pattern, indicating poor cell growth activities in 2-D matrigel. Tumor growth in nude mice in the C + A + P group was inhibited (P<0.01).

CONCLUSIONshRNA targeting COX-2, AKT1 and PIK3R1 down regulated significantly the expression of the three genes in a sequence-specific manner, exerted proliferation inhibition effect on SGC7901 cells in vitro and in vivo.

Adenocarcinoma ; genetics ; physiopathology ; therapy ; Adenoviridae ; genetics ; metabolism ; Animals ; Cell Line, Tumor ; Cell Proliferation ; Cyclooxygenase 2 ; genetics ; metabolism ; Disease Models, Animal ; Down-Regulation ; Genetic Therapy ; Genetic Vectors ; genetics ; metabolism ; Humans ; Inverted Repeat Sequences ; Mice ; Mice, Nude ; Phosphatidylinositol 3-Kinases ; genetics ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; RNA Interference ; RNA, Small Interfering ; genetics ; therapeutic use ; Stomach Neoplasms ; genetics ; physiopathology ; therapy