This study aims to optimize the most effective component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on lung cancer A549 using the orthogonal design method, and to investigate its effects of the component formula on cell proliferation, apoptosis and cytoskeleton in lung cancer A549 cells. The orthogonal design method was introduced to optimize the most effective component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on lung cancer A549 cells. CCK-8 assay and Real-time cell analysis were adapted to analyze the effect of component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on A549 cells viability at different time and dose. Cell apoptosis was measured by Annexin V- FITC/PI double staining and flow cytometry. Cell skeleton protein F-actin was detected by high content screening (HCS). The optimizing component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma for total salvianolic acid, total saponins of panax ginseng and ginseng polysaccharide doses were 5, 10, 5 mg L(-1). CCK-8 assay and real-time cell analysis demonstrated that the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma treatment could significantly decrease the A549 cell viability in both dose- and time-dependent manner compared with control group (P < 0.01). Moreover, the increase of cell apoptosis was detected by Annexin V-FITC/PI double staining and flow cytometry when cells treated with the component formula, which indicating that the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma could induce A549 cell apoptosis in a time-dependent manner compared with control group (P < 0.01). Furthermore, compared with control group, a significant decrease in A549 cell skeleton area was found in the component formula-exposed cells in the dose-dependent manner (P < 0.01). In summary, the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma inhibits A549 cell proliferation by inducing cell apoptosis and decreasing cell microfilament formation. All of these results will be helpful to reveal antitumor mechanism of the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma, which provides a basis for the exploration of antitumor mechanism of the component formula on lung cancer.
Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Lung Neoplasms ; drug therapy ; Panax ; chemistry ; Plant Extracts ; chemistry ; pharmacology ; Plant Roots ; chemistry ; Rhizome ; chemistry ; Salvia miltiorrhiza ; chemistry

Objective To study the effects of oxymatrine as inhibitor of hemorrhagic fever with renal syndrome virus (HFRSV) infection in vitro and in vivo.Methods In vitro studies,a dose of oxymatrine without cytotoxicity and 76-118 strain of HFRSV was taken to treat Vero cells in three ways:①After treated with oxymatrine for 48 h,Vero cells were attacked by HFRSV at dilution of 10-1 ～ 10-6,respectively for 24 h before changing to maintenance medium; ②Vero cells were first attacked by HFRSV of 10-1 ～ 10-6 dilution respectively,then oxymatrine was used for 48 h before changing to maintenance medium; ③Vero cells were attacked by HFRSV at dilution of 10-1 ～ 10-6 respectively,and meanwhile treated with oxymatrine for 48 h before changing to maintenance mcdium.Each dilution handled four porocytes,and four positive controls were set up at the same time.Indirect immunofluorescence assay (IFA) was performed to determine the inhibitory effect of oxymatrine in experimental group and positive control.In vivo studies,thirty 2-week-old hamsters,weighing about 30-40 g,were divided into experimental and control groups according to body weight,n =15.These aninals were inoculated intraperitoneally with HFRSV in 100TCID50(0.1 ml each); on days 4-13,0.1 ml of oxymatrine 1:100 were given to each hamster in experimental group daily by intraperitoneal injection,while the same amount of saline was given to the control ones.Lung tissue of hamsters was then dissected out to slice to be identified by immunofluorcscence stain.Results It was demonstrated that oxymatrine with the diluted fractions of 1:8 was safe in vitro.When the virus dilution of HFRSV was l0-4,compared with control groups,the differences were statistically significant in method 2 and 3 (z =-2.53,-2.53,all P ＜ 0.05),while no statistical significance in method 1 (z＝5.36,P＞ 0.05).When the virus dilution of HFRSV was 10-1 ～ 10-3,10-5,10-6,the differences were not statistically significant (z--0.00,-0.32,-0.19,4.21,4.21,all P ＞ 0.05).In vivo studies,compared with control group,the differences were statistically significant in experimental group (z =-3.85,P ＜ 0.05).Conclusion Oxymatrine significantly inhibites HFRSV.

Objective To investigate the copy number changes on chromosome 3q26. 1 in urothelial carcinoma of the bladder, and to explore its potential clinical significance. Methods The microarray-based comparative genomic hybridization (Array-CGH) approach was used to analyze the genome-wide copy number changes of 35 tumor tissue samples of bladder cancer. To confirm the loss of a small fragment in 3q26. 1 detected by Array-CGH, real-time fluorescent quantitative polymerase chain reaction (real-time PCR) was performed with 57 frozen tumor tissue samples and 34 formalinfixed paraffin-embedded (FFPE) tumor tissue samples. The urine sediment cells collected from 15 healthy volunteers and 29 bladder cancer patients were checked as above. Results The Array-CGH data showed that the copy number loss of a small fragment in 3q26. 1 was detected in 77.1% (27/35)of the tumor tissue samples investigated. Real-time PCR analysis validated this loss of a small fragment of 3q26.1 with high frequencies in both 57 frozen tumor samples and 34 FFPE tumor samples.The percentage of samples exhibiting loss was 78.9% (45/57) and 100. 0% (34/34) respectively.Furthermore, the relative copy number of the 3q26.1 small fragment was significantly lower in the urinary sediment cells of the patients (median=0. 0020), comparing with that of healthy controls (median=0. 0030) (P＜0.01). Conclusions Loss of the small fragment in 3q26.1 could be a characteristic genetic change of urothelial carcinoma of the bladder. It may serve as a potential molecular marker for bladder cancer.

BACKGROUNDTarget-controlled infusion (TCI) has been recently developed and successfully implemented in clinical practice. The current study was to estimate the population pharmacokinetics of rocuronium TCI in adult patients using nonlinear mixed-effects model (NONMEM), and to investigate the influence of relevant factors in adult patients.

METHODSFourteen ASA I-II patients undergoing elective laparoscopy operation with general anesthesia were included. After induction, all patients received rocuronium by TCI system. The beginning target plasma concentration (Cpt) was 2.0 µg/ml, then increased Cpt according to the neuromuscular transmission monitoring. The endpoint of Cpt was determined when the T₁ scale was blocked by 90% - 95%. TCI rocuronium was stopped 30 minutes before the end of the operation. Arterial blood was drawn before anesthesia at 0, 2, 4, 6, 8, 10, 15, 20, 30, 45, 60, 120, 180, 240 and 360 minutes after the infusion of rocuronium was stopped for the analysis of plasma concentrations of rocuronium by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). The population pharmacokinetics analysis was performed using NONMEM program.

RESULTSThe pharmacokinetics of TCI rocuronium in adult patients was best described by a three-compartment model. Pharmacokinetic parameters were clearance (CL)₁ = 0.205 L/min, CL₂ = 0.324 L/min, CL₃ = 0.0292 L/min, volumes of distribution (V)₁ = 4.00 L, V₂ = 2.28 L, V₃ = 4.26 L, Vdss = 10.54 L. Both age and weight as covariates affected the pharmacokinetic parameters. V₁ and CL₁ were negatively correlated with patient age. CL₁ was positively correlated with weight.

CONCLUSIONSNo pharmacokinetic change was noted when rocuronium was administered via TCI. Both age and weight as covariates affected the pharmacokinetic parameters.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Androstanols ; administration & dosage ; pharmacokinetics ; Female ; Humans ; Infusion Pumps ; Male ; Middle Aged ; Young Adult

Objective To study the clinical characteristics of bulbar myasthenia gravis (MG). Methods Retrospective review was performed on 166 patients with bulbar type of myasthenia gravis, diagnosed at Tongji Hospital in the period of May 1983 through October 2005.Results Bulbar MG was a relatively rare type of MG,accounting for 5.7% (166/2888) of MG classifications.Females were more often affected than males (the ratio of male:female was 1:1.35).The peak of onset age was at 20—40 years.The incidence of myasthenia crisis in the group was 26.5% (44/166).Myasthenic crisis occurred in 10.8% (18/166) of the bulbar MG patients within 6 months after onset,resulting in a mortality rate of 6.0% (10/166) in the group.Out of the group,30 cases experienced puhnonary infections (18.1%). Thirty cases were initially misdiagnosed as other diseases such as nasopharyngeal disorders (33/166, 19.9%).The routine therapy was not very satisfactory.Median dose cyclophosphamide therapy appeared to be effective for ameliorating refractory MG.Thymectomy was performed in 25 patients,with optimistic efficacy rate up to 80.0% (20/25) in a 3-year follow-up.Conclusions The clinical analysis in the current study suggested that the bulbar MG had its own characteristics in such aspects as progression of the disease, complications,treatment and prognosis.The information of the clinical manifestations presented in this study may be useful in diagnosing and treating bulbar MG.

Objective To explore the effect of low frequency repetitive transranial magnetic stimulation (rTMS) on unilateral spatial neglect (USN). Methods 40 stroke patients with USN were divided into treatment group (n=20) and control group (n=20). Patients in the treatment group were treated with low frequency rTMS for 2 weeks. The USN degree of these groups were evaluated before and after the treatment. Results There was no significant difference of USN degree between these groups before the treatment (P>0.05); Compared with the control group, the treatment group improved significantly after the treatment (P<0.05). The USN degree of patients in the treatment group decreased significantly after the treatment (P<0.05), while patients in the controlled group had no difference (P>0.05). Conclusion USN induced by stroke could be improved obviously through low frequency rTMS.

BACKGROUNDRecently, the combination of sevoflurane and remifentanil has been widely used in general anesthesia. In this study, we investigated the sevoflurane-remifentanil pharmacodynamic interactions at clinical concentrations using the observer's assessment of alertness/sedation (OAA/S) and the bispectral index (BIS) by response surface analysis.

METHODSTotally 65 American Society of Anesthesiologists (ASA) I patients age 20 to 50 years old were included in this study. Patients were randomly assigned to be anesthetized with different target end-tidal sevoflurane concentrations that ranged from 0.2% to 3.4% in increments of 0.2%. The end-tidal sevoflurane concentration was maintained constant throughout the study. Remifentanil was infused with a target controlled infusion (TCI) system at increasing step-wise concentrations from 1 ng/ml to 10 ng/ml. The values of OAA/S and BIS at different sevoflurane-remifentanil concentration combinations were measured. The pharmacodynamic interactions between sevoflurane and remifentanil were analyzed by a response surface method. The three-dimensional response surfaces were constructed with Minitab Software. Model parameters were estimated with NONMEM program.

RESULTSSevoflurane and remifentanil acted synergistically on OAA/S. Sevoflurane alone could produce OAA/S ≤ 1 at a minimal alveolar concentration (MAC) of 0.93%. When used in combination with remifentanil at 1, 3, 6, and 10 ng/ml, the corresponding sevoflurane MACs were reduced to 0.79%, 0.58%, 0.48%, and 0.38%, with reductions of 17.2%, 37.6%, 48.4%, and 62.0% from baseline, respectively. In patients administered remifentanil alone, the OAA/S score was ≥ 3 even when the remifentanil concentration reached 10 ng/ml. BIS was closely associated with the sevoflurane concentration and the remifentanil concentration did not noticeably influence the relationship between the sevoflurane concentration and BIS. A sevoflurane concentration of (1.04 ± 0.19)% to (1.81 ± 0.21)% could maintain a BIS between 60 and 40.

CONCLUSIONSThe response surface method can analyze the pharmacodynamic interactions between remifentanil and sevoflurane qualitatively and quantitatively. Within the range of our study (remifentanil ≤ 10 ng/ml, sevoflurane ≤ 3.4%), the two drugs produced synergistic effects on OAA/S but had no interactive effect on BIS. A guideline of BIS between 40 and 60 may cause excessive anesthesia when opioids are used to maintain anesthesia.

Adult ; Anesthesia ; methods ; Consciousness ; drug effects ; Female ; Humans ; Male ; Methyl Ethers ; pharmacokinetics ; pharmacology ; Middle Aged ; Piperidines ; pharmacokinetics ; pharmacology ; Young Adult

Animals ; Disease Models, Animal ; Liver ; pathology ; Liver Failure, Acute ; metabolism ; pathology ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Transcription Factor RelA ; metabolism

As a novel and effective approach, response surface model is used in the study of drug-drug interactions. When two drugs are used simultaneously, this model can be applied to estimate the key characters of the response surface, to find the desired response region by optimal drugs combination, to explore the mechanism of drug-drug interactions, and thus to guide sound clinical application and reduce the risk and cost. In this article, the model's basic mathematical and pharmacological concepts are introduced, and its research progresses are reviewed.
Alfentanil ; pharmacology ; Anesthetics ; pharmacology ; Computer Simulation ; Drug Interactions ; Drug Synergism ; Humans ; Midazolam ; pharmacology ; Models, Statistical ; Propofol ; pharmacology

BACKGROUNDLittle is known about the influence of liver transplantation on the pharmacokinetics of most anesthetic drugs. The goal of this study was to study the population pharmacokinetics of remifentanil in the different phases of orthotopic liver transplantation (OLT) and the influence of relevant factors.

METHODSThirteen adult patients undergoing OLT were enrolled. A single bolus infusion of remifentanil 5 microg/kg was administered during the preanhepatic, anhepatic and neohepatic phases of OLT. Arterial blood samples of 1.5 ml were collected at 0 (baseline), 1, 2, 3, 5, 7, 10, 15, 20, 25, 30, 45, 60 and 90 minutes after drug administration. Remifentanil concentration was assayed by high-performance liquid chromatography/mass spectrometry/mass spectrometry (HPLC/MS/MS). Population pharmacokinetic modeling was performed using nonlinear mixed-effects modeling (NONMEM).

RESULTSThe pharmacokinetics of remifentanil in patients undergoing OLT was best described by a two-compartment open model. The pharmacokinetic parameters were not influenced by age, gender, operative phase, blood temperature, rehydration volume, or blood loss volume during sampling. The volume of distribution in the central compartment (V(1)) and the volume of distribution in the peripheral compartment (V(2)) were influenced by body weight.

CONCLUSIONSThe population pharmacokinetics of remifentanil in patients undergoing OLT can be well described by a two-compartment open model. The functional status of the liver does not significantly affect the pharmacokinetics of remifentanil, but the body weight is an influential factor of V(1) and V(2).

Adult ; Chromatography, High Pressure Liquid ; Female ; Humans ; Liver Transplantation ; Male ; Middle Aged ; Piperidines ; administration & dosage ; pharmacokinetics ; Tandem Mass Spectrometry