Objective To investigate the association between the expression of heparanase(Hpa) and the invasion of choriocarcinoma by studying the expression of Hpa in human choriocarcinoma cell lines JEG-3 and JAR and human chorionic villous tissues.Methods(1)Matrigcl invasion assays were used to detect in vitro invasive ability of JEG-3 cells and JAR cells.(2)Expression of Hpa protein in the human chorionic villous tissues and choriocarcinoma cell lines(JEG-3 cells and JAR cells)were detected by immunocytochemistry and western blot.Results(1)The invasive cell number was significantly larger in JEG-3 cells than in JAR cells(191?17 vs 106?13,P

OBJECTIVEThe aim of the present study was to prepare uniform-sized silybin loaded poly (lactic-co-glycolic acid) (PLGA) microspheres in study of silybin with stainless steel membrane.

METHODSilybin PLGA microspheres were prepared by stainless steel membrane emulsification. The preparation conditions were optimized by single-factor test and orthogonal experiment, and evaluating the mean diameters, the particle size distribution, drug loading, entrapment efficiency and morphology of microsphere.

RESULTPrepared microspheres were round and surface was smooth. The mean diameter was (4.961 +/- 0.56) microm. The span was (1.75 +/- 0.18). The entrapment efficiency was (54.997 +/- 4.05)% and the average drug loading was (23.6 +/- 1.70)%.

CONCLUSIONThe stainless steel membrane emulsification can be used to prepare the silybin PLGA microspheres. The mean diameters of the silybin PLGA microspheres can be controlled in certain level. Stainless steel membrane emulsification has great potentiality exploitation and utilization.

Drug Compounding ; methods ; Emulsions ; chemistry ; Lactic Acid ; chemistry ; Microspheres ; Particle Size ; Polyglycolic Acid ; chemistry ; Silymarin ; chemistry ; Stainless Steel ; chemistry

Objective To discuss the occurrence of anxiety in subarachnoid hemorrhage (SAH) patients and the analysis of correlation factors. Methods The occurrences of anxiety in 73 SAH patients were studied with a questionnaire. The correlations between the anxiety occurrence and the gender, age, education state, mode of medicine fee payment, SAH times, clinical symptoms, knowledge of the disease and complications were analyzed. Results The occurrence rate of anxiety in SAH patients was 53.4%. The female patients had a higher occurrence rate of anxiety than that of the males. Severe headache patients had a higher rate of anxiety than the slight ones. Hemorrhages times and complications had Obvious correlations with anxiety. Conclusions Correct understanding of anxiety is the precondition of expanding the intervention of positive emotion. Analyzing the anxiety-related factors and then actively giving the physiological therapy are the keys to the successful operation in SAH patients.

BACKGROUNDIn China, tracheobronchial foreign body (TFB) aspiration, a major cause of emergency episode and accident death in children, remains a challenge for anesthetic management. Here, we share our experience and discuss the anesthetic consideration and management of patients with TFB aspiration.

METHODSThis was a single-institution retrospective study in children with an inhaled foreign body between 1991 and 2010 that focused on the complications following rigid bronchoscopy (RB). Data including the clinical characteristics of patients and TFB, anesthetic method, and postoperative severe complications were analyzed by different periods.

RESULTSDuring the 20-year study period, the charts of 3149 patients who underwent RB for suspected inhaled TFB were reviewed. There were 2079 male and 1070 female patients (1.94:1). A nut (84%) was the most commonly inhaled object. The study revealed a 9% (n = 284) overall rate of severe postoperative complications related to severe hypoxemia, laryngeal edema, complete laryngospasm, pneumothorax, total segmental atelectasis, and death with incidences of 3.2%, 0.9%, 1.3%, 0.3%, 0.3%, and 0.1%, respectively. The rates of preoperative airway impairment, negative findings of TFB, and adverse postoperative events have been on the rise in the past 5 years.

CONCLUSIONSThe survey results confirmed that hypoxemia remains the most common postoperative complication in different periods. Both controlled ventilation and spontaneous ventilation were effective during the RB extraction of the foreign body at our hospital in the modern technique period. An active respiratory symptom was commonly seen in the groups with negative findings.

Airway Obstruction ; complications ; diagnosis ; epidemiology ; surgery ; Bronchoscopy ; adverse effects ; Child ; Child, Preschool ; Female ; Foreign Bodies ; Humans ; Hypoxia ; diagnosis ; epidemiology ; etiology ; Male ; Retrospective Studies

Objective To detect carbapenemase production in enterobacteriaceae by using modified Hedge test (MHT) and investigate the distribution of carbapenemase genes.Methods A total of 3 718 isolates from enterobacteriaceae were collected from 4 hospitals,including Second Affiliated Hospital of Zhejiang University,People's Hospital of Dongyang,Beijing Hospital,and Huaxi Hospital of Sichuan University.Susceptibility of enterobacteriaceae to ertapenem was tested by K-B method.MHT was used to detect carbapenemase in enterobacteriaceae and the common carbapenemase genes were amplified by using PCR.Results The total resistance rate ( resistance and intermediate resistance) of enterobacteriaceae to ertapenem was 3.04％ (113/3718) and there were differences in resistance rate of enterobacteriaceae to ertapenem among 4 hospitals (5.09％,2.15％,2.59％,and 1.72％,respectively).Of the 3718 isolates,2.29％ (85) were positive to MHT,and there were differences in positive rate to MHT among 4 hospitals (4.73％,1.21％,1.06％,and 1.58％,respectivdy).Of 113 non-susceptible isolates to ertapenem,82 were positive to MHT and 31 were negative.Of 85 MHT-positive isolates,82 were resistant or intermediate resistant to ertapenem and only 3 were susceptible.Of 82 MHT-pesitive and non-susceptible isolates to ertapenem,65 were positive to KPC gene,15 were positive to IMP gene (two of them were positive to both KPC and IMP),and 4 were negative to all tested carbapenemase genes.Thirty-one MHT-negative and nonsusceptible to ertapenem and 3 MHT-positive and susceptible isolates to ertapenem were negative to all tested carbapenemase genes.Conclusions MHT is used to detect carbapenemase in enterobacteriaceae with high sensitivity and low false positive rate.KPC gene is the most occurred gene to be dominant in the production of carbapenemase in enterobacteriaceae, and the IMP gene is also responsible to the genesis ofcarbapenemase in enterobacteriaceae..

Case-Control Studies ; Creatine ; urine ; Female ; Humans ; Hypoxia-Ischemia, Brain ; diagnosis ; urine ; Infant, Newborn ; Lactic Acid ; urine ; Male ; Predictive Value of Tests

The high expression of tissue factor (TF) is related to the coagulation disorder in acute leukemia. TF in blood circulation is mainly expressed in cells, microparticles (MP) and alternatively spliced human tissue factor (asHTF). To elucidate the role of TF in the coagulation disorder of acute myeloid leukemia (AML), RT-PCR was performed on 6 common AML cell lines NB4, HL-60, Kasumi-1, U937, K562 and THP-1. The results showed that only NB4 and U937 cells expressed baseline full-length TF and asHTF which were proved by sequencing. The flow cytometric detection, TF activity and TF antigen tests in NB4 and U937 cells revealed that the asHTF was expressed in trace amount and almost had no activity, while the TF antigen and activity in microparticles were significantly higher than that in asHTF. It is concluded that asHTF may play an unimportant role in the coagulation disorder of AML. Microparticle associated tissue factor (MP-TF) is the predominant source of TF activity released from AML cells.
Alternative Splicing ; HL-60 Cells ; Humans ; Leukemia, Promyelocytic, Acute ; genetics ; metabolism ; Thromboplastin ; genetics ; metabolism ; Tumor Cells, Cultured ; U937 Cells

BACKGROUNDThe evoked electromyography (EMG) is frequently used to identify facial nerve in order to prevent its damage during surgeries. Partial neuromuscular blockade (NMB) has been suggested to favor EMG activity and insure patients' safety. The aim of this study was to determine an adequate level of NMB correspondent to sensible facial nerve identification by evaluating the relationship between facial EMG responses and peripheral NMB levels during the middle ear surgeries.

METHODSFacial nerve evoked EMG and NMB monitoring were performed simultaneously in 40 patients who underwent tympanoplasty. Facial electromyographic responses were recorded by insertion of needle electrodes into the orbicularis oris and orbicularis oculi muscles after electrical stimulation on facial nerve. The NMB was observed objectively with the hypothenar muscle's twitching after electrical stimulation of ulnar nerve, and the intensity of blockade was adjusted at levels of 0, 25%, 50%, 75%, 90%, and 100% respectively with increased intravenous infusion of Rocuronium (muscle relaxant).

RESULTSAll of the patients had detectable EMG responses at the levels of NMB or= 75%. A significant linear positive correlation was present between stimulation thresholds and NMB levels while a linear negative correlation was present between EMG amplitudes and NMB levels.

CONCLUSIONSThe facial nerve monitoring via facial electromyographic responses can be obtained when an intraoperative partial neuromuscular blockade is induced to provide an adequate immobilization of the patient. The 50% NMB should be considered as the choice of anesthetic management for facial nerve monitoring in otologic microsurgery based on the relationship of correlation.

Adult ; Ear, Middle ; surgery ; Electromyography ; methods ; Facial Nerve ; physiology ; Female ; Humans ; Male ; Middle Aged ; Monitoring, Intraoperative ; methods ; Neuromuscular Blockade ; methods ; Otologic Surgical Procedures ; methods ; Young Adult

OBJECTIVETo prepare the polyclonal antibody of human endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1), and to determine the expression of EOLA1 in human umbilical vein endothelial cell (HUVEC).

METHODSThe protein samples (sample 1 and 2) expressing EOLA1 were purified and renatured. The protein concentrations were determined with bicinchoninic acid assay. The protein samples were identified with peptide mass fingerprinting (PMF) analysis. Protein sample with higher coincidence rate of amino acid sequence with theoretic protein was used to inoculate 4 mice; another 4 mice inoculated with adjuvant were used as control. Serum was isolated from collected mice blood. Polyclonal antibody of EOLA1 was purified with saturated ammonium sulfate precipitation, and was determined with ELISA for the titer (data were denoted by absorbance value). The expression of EOLA1 in HUVEC was determined with Western blot.

RESULTSThe concentration of protein sample 1 and 2 was respectively 0.124 16 mg/mL and 0.132 15 mg/mL. According to PMF analysis, the coincidence rate of amino acid sequence between protein samples and theoretic protein were 32% (protein sample 1) and 24% (protein sample 2). The polyclonal antibody of EOLA1 with titer more than 1:10 000 was obtained from mice inoculated with protein sample 1. The expression of EOLA1 protein in HUVEC was determined with polyclonal antibody of EOLA1.

CONCLUSIONSThe polyclonal antibody of EOLA1 can be prepared by inoculating mice with EOLA1 prokaryotic expressing protein, which can be used for determination of EOLA1 protein.

Animals ; Antibodies ; Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; metabolism ; Humans ; Lipopolysaccharides ; metabolism ; Membrane Proteins ; immunology ; metabolism ; Mice

OBJECTIVETo investigate the essential role and mechanism of TRPC6 gene in the development of gastric cancer.

METHODSThe expression of TRPC6 protein was assessed in gastric cancer tissues and normal tissues adjacent to the cancer from 30 patients with gastric cancer. The inhibiting effect of TRPC6 activity on cell growth, cell cycle of a human gastric cancer cell line AGS cells, tumor progression and development of xenografted human gastric cancer in a mouse model was tested using dominant-negative mutant TRPC6 (DNC6). The survival of mice bearing xenografted tumors in the GFP and DNC6 was compared using Kaplan-Meier analysis. All statistical tests were two-sided.

RESULTSThe TRPC6 protein in the tumor tissues and para-tumor tissues was (21.60 ± 8.32)% versus (7.14 ± 2.24)%. After transfection of DNC6 virus for 24 hours, 48 hours, 72 hours and 96 hours, the growth inhibition rates of gastric cancer cells were (36.90 ± 1.13)%, (44.06 ± 2.17)%, (52.12 ± 2.76)% and (50.89 ± 1.97)%, respectively. The clone formation rates of control group and DNC6 group were (14.70 ± 3.00)% versus (43.80 ± 7.00)%. After transfection with DNC6 virus for 0, 24, 36 and 48 hours, the G(2)/M phase arrest was (20.34 ± 1.98)%, (24.31 ± 2.37)%, (27.70 ± 2.36)%, (35.10 ± 3.0)% in the DNC6 group and (18.40 ± 2.01)%, (18.0% ± 1.72)%, (17.50 ± 1.74)%, (16.80 ± 1.71)% in the control group, respectively. Inhibition of TRPC6 activity also reduced the subcutaneous tumor volume in the mouse models with xenografted human tumors (P < 0.05).

CONCLUSIONIn the preclinical models tested, TRPC6 channels are essential for gastric cancer development via regulation of G(2)/M phase transition.

Adenoviridae ; genetics ; Animals ; CDC2 Protein Kinase ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Cyclin B ; metabolism ; Cyclin-Dependent Kinases ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Recombinant Proteins ; metabolism ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; TRPC Cation Channels ; metabolism ; TRPC6 Cation Channel ; Transfection ; Tumor Burden