Child, Preschool ; Echocardiography, Transesophageal ; methods ; Heart Septal Defects, Ventricular ; diagnostic imaging ; therapy ; Humans ; Male

Aim To establish an in vitro early drug cardiac tox-icity evaluation method by human embryonic stem cells derived cardiomyocytes ( hESC-CM) and real-time cell analysis Cardio (RTCA Cardio) system. Method The hESC-CM were cultured at RTCA Cardio E-Plate 96. Impedance signals from hESC-CM were analyzed for beating rate, contraction amplitude and beating rhythm irregularity to determine the optimum inoculation density and detection duration. Based on this, we used 0. 1 % DMSO to be the solvent and quinidine (0. 2, 0. 78, 3. 13, 12. 5, 50 and 100 μmol·L - 1 ) known as affecting cardiac activity to validate this method. Result The results revealed no significant changes in the cell index (CI), transient pulse patterns, beating rate and amplitude of hESC-CM. Quinidine will affect the CI and transi-ent pulse patterns of hESC-CM and decrease the beating rate and amplitude of hESC-CM when its concentration ≥3. 13 μmol · L - 1 . And this effect is concentration-dependent, the higher the concentration,the more time they need to recover beating and the more significant the beating rate and amplitude inhibition of quinidine on hESC-CM. Conclusion The method established by hESC-CM and RTCA Cardio system can detect the effect of quinidine on the contraction of hESC-CM, and this indicates that this method has the potential to be an attractive high-throughput tool for screening potential drugs in early evaluation of drug car-diotoxicity.

Objective To explore the influence of Akt inhibitor MK2206 in the proliferation and apoptosis of tongue squamous cell carcinoma TCA-8113 cells,and to clarify the possible mechanism.Methods The tongue squamous carcinoma TCA-8113 cells at the logarithmic phase were randomly divided into control group and 1,5,25,125, 250 nmol·L-1 MK2206 groups.The inhibitory rate of proliferation of TCA-8113 cells was detected with MTT method,and the apoptotic rate of TCA-8113 cells was determined with flow cytometry(FCM),and the expressions of caspase-9,Bad,GSK-3β,p-Akt and T-Akt proteins in the TCA-8113 cells were detected with Western blotting method.Results The IC50 of tongue squamous cell carcinoma TCA-8113 cells after treated with MK2206 for 12, 24,and 36 h were (112.54±1.67),(79.67±2.01),and (33.33±1.98)nmol·L-1 .The FCM results showed that the apoptotic rates of TCA-8113 cells after treated with 1,5,25,125,and 250 nmol·L-1 MK2206 for 12 h were (14.2±0.74)%,(19.3±0.45)%,(35.1±0.45)%,(39.6±0.48)% and (52.1±0.19)%;there were significant differences compared with control group(P<0.01).The Western blotting method results showed that the expressions of p-Akt, Bad and GSK-3βwere decreased with the increasing of dose and time of MK2206;compared with theβ-actin in control group,the bands got darken;the expression level of caspase-9 was increased, compared with theβ-actin in control group, the bands got darken;the T-Akt protein expression did not change significantly;compared with the β-actin in control group, the color of bands had no significant difference.Conclusion Akt inhibitor MK2206 can inhibit the proliferation of tongue squamous cell carcinoma TCA-8113 cells and induce apoptosis.

Objective To discusse the influence of permeating perfluoropentane drops-encapsulated mesoporous silica nanocapsules (MSNC-PFP) on HIFU surface ablation in vitro bovine liver.Methods Fifity bovine liver tissues were divided into 5 groups:Control group,0.25 mg/ml group,0.50 mg/ml group,1.00 mg/ml group,2.00 mg/ml group according to the concentration of MSNC-PFP.The synergistic agent was injected along the ablation line path under the guiding of ultrasound.The surface ablation model was structured by 6 coagulation necrosis side by using HIFU linear scanning,inner region without melting,and the ultrasonographic changes were observed in the melt area.The coagulation necrosis range was assessed by TTC staining,and necrosis degree was observed by HE staining.The coagulation necrosis volume was measured,the coverage index (CI),external volume index (EI),energy efficiency factor (EEF) were used to evaluate the ablation effect in each group.Results The MSNC-PFP more than 1.00 mg/ml was shown as strong echo in ultrasonic image,fade after 3-5 min.The grey value was changed in HIFU ablation area related to the concentration of MSNC-PFP.The total ablation volume of 0.50 mg/ml group,1.00 mg/ml group,2.00 mg/ml group were more than control group,the EEF was lower than that of the control group (all P＜0.01).The CI and EI of 1.00 mg/ml group and 2.00 mg/ml group were higher than those of the other group (all P＜0.01).The coagulation necrosis became complete when the concentration of MSNC-PFP above 0.50 mg/ml.Conclusion MSNC-PFP can increase the volume of coagulation necrosis,decrease the EEF and increase the efficiency of HIFU surface ablation of bovine liver in vitro.

Objective To study the method and efficacy of double chemoembolization of hepatic carcinoma by transcatheter arterial cheomembolization (TACE) and portal vein port catheter system (PCS). Methods PCS was implanted in portal vein in 17 cases of irresectable primary hepatocellular cancer (HCC) during operation. One month later, chemoembolization of portal vein via PCS was performed, meanwhile TACE was also done (hepatic arterial portable vein chemoembolization, HA PVCE). A total of 120 cases of HCC treated simply with TACE were served as the control. Results The technical success rate was 100% in both TACE and portal vein PCS. Hepatic arteriography showed rich or moderate blood supply of HCC in 134 cases and PCS portography showed tumor staining in 6 cases. In groups of HA PVCE and TACE, the positive alpha fetoprotein (AFP) level decreased in 80% and 71.6% cases; half a year survival rate was 85.7% and 74.4%, and one year survival rate was 71.4% and 52.4%, respectively. Conclusion Double chemoembolization by TACE and portal vein PCS is a safe procedure with high technique success rate and therapeutic efficacy for irresectable hepatic carcinoma.

Tanshinones, the main bioactive compounds of Salvia miltiorrhiza, are the diterpenoid pigments, multiple genes were proved to be involved in their biosynthesis in plants. CYP76AH1 is the initial P450 gene in the tanshinones biosynthetic pathway, its function has been validated by yeast expression and in vitroenzymatic reaction. In order to clarify the function of CYP76AH1 in vivo, in this study, we constructedthe RNA interference of CYP7AH1 in S. miltiorrhiza hairy root. The RNA interference vector with a hairpin structure was constructed using the Gateway technology, and then the interference fragment was integrated into the genome of S. miltiorrhiza mediated by Agrobacterium rhizogenes. Several highly CYP76AH1 interference S. miltiorrhiza hairy roots were obtained for further analysis.
Agrobacterium ; genetics ; metabolism ; Biosynthetic Pathways ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Diterpenes, Abietane ; biosynthesis ; Gene Expression Regulation, Plant ; Plant Proteins ; genetics ; metabolism ; RNA Interference ; Salvia miltiorrhiza ; genetics ; metabolism ; microbiology

OBJECTIVETo improve the method for monitoring sleep state and respiratory rhythm of SD rats, providing a solution for rats' chewing on the wires, signal loss and instability problems in the animal model of sleep apnea syndrome (SAS).

METHODSWe improved monitoring electrodes of both electrocorticogram (ECoG) and electromyogram (EMG), signal circuit and animal operation.

RESULTSOperation time was shortened and wound exposure time was reduced, which made it easier for postoperative recovery. The ECoG and EMG signals were more stable with sharp image, and signal circuit lines had better conductivity and material durability, achieving continuous monitoring for a long time and high success rate. We could precisely distinguish the sleep wake state and the sleep apnea events in rats according to these signals.

CONCLUSIONThe improved method is more reliable and practical to test the small animal model of SAS, and is more easily to operate and the signals are more stable.

Animals ; Electroencephalography ; methods ; Electromyography ; methods ; Models, Animal ; Monitoring, Physiologic ; methods ; Rats ; Respiration ; Sleep ; Sleep Apnea Syndromes ; diagnosis

Objective To optimize immunodominant protein combinations for serological screening for Cblamydia trachomatis (Ct) infection.Methods Both serum and genital swab samples were collected from 50 patients with Ct infection confirmed by colloidal gold immunochromatographic assay (GICA),and 30 GICA-negative clients without Ct infection at a sexually transmitted disease (STD) clinic in Tianjin Medical University General Hospital.The 30 serum samples from GICA-negative clients were also negative for microimmunofluorescence (MIF) assay.Eight Ct immunodominant proteins,including Pgp3,CPAF,CT143,CT101,CT694,CT875,CT813 and IncA,were selected as antigens to detect corresponding antibodies in the serum samples by enzyme-linked immunosorbent assay (ELISA) with the Ct proteins Hsp60 and major outer membrane protein (MOMP) as references.The results of ELISA were compared with those of the traditional gold standard method MIF assay to determine the immunodominant protein combination with the highest sensitivity and specificity.Results Of the 50 serum samples from patients with Ct infection,44 were positive and 6 negative by MIF.The results of ELISA with the combination of immunodominant proteins Pgp3,CT694 and CT875 as antigens were 97.73％ (43/44) consistent to those of MIF assay.Of the 30 serum samples from GICA-negative clients,all were negative by MIF.Meanwhile,no antibody was detected against any of the immunodominant proteins Pgp3,CT694 and CT875 in any of the serum samples from GICA-negative clients.Conclusions The ELISA with the combination of immunodominant proteins Pgp3,CT694 and CT875 as antigens has good sensitivity and specificity for serological screening for Ct infection,and is simple to operate and easy to popularize.

BACKGROUND:In the treatment of tibial plateau fractures, because of the variety of fracture, the complexity of anatomical changes, X-ray films or three-dimensional CT scan limited by two-dimensional plane, increases the difficulty in preoperative plan and surgical treatment. The application of three-dimensional (3D) printing technology has attracted attention in the department of orthopedics. OBJECTIVE:To explore the auxiliary role of 3D printing technique in preoperative plan and treatment for tibial plateau fractures. METHODS:Thirty patients with tibial plateau comminuted fractures were enroled in this study and divided into two groups: experimental and control groups, with 15 patients in each group. In the experimental group, patients underwent 3D CT scan, which was stored in DICOM format, and processed by Mimics software. Data were converted into STL format, entered 3D printer, and a 1:1 entity size of the fracture model was made, in accordance with repair plan of 3D fracture model. Operation time and intraoperative blood loss were compared between the two groups. At 12 months after treatment, their outcomes were assessed using Rasmussen evaluation criteria. RESULTS AND CONCLUSION: The 3D printing fracture models of 1:1 ratio identified fracture type and made a repair program before surgery in the experimental group. Operation time and intraoperative blood loss were significantly less in the experimental group than in the control group (P < 0.05). After surgery, patients were folowed up for 12 to 18 months. The healing time was 3-5 months, averagely 4.3 months. At 12 months after treatment, the Rasmussen evaluation criteria results showed that the excelent and good rate was significantly higher in the experimental group than in the control group (P < 0.05). These results suggest that the fracture model of 3D can help to make the operation plan. The treatment of tibial plateau fractures is more precise, personalized and visual.

OBJECTIVE: To investigate the signal transduction mechanism of curcumin in inhibiting the proliferation of bovine lens epithelial cell (LEC) induced by recombinant human epidermal growth factor (rhEGF). METHODS: There were three groups in this experiment, which were normal control group, untreated group and curcumin-treated group. Proliferation of LEC was induced by rhEGF (50 microg/L). The concentration of intracellular Ca(2+) ([Ca(2+)](i)) in LEC was measured with Fura-2/AM by spectrofluorimetry. The contents of intracellular cAMP and cGMP were assayed by radioimmunoassay. RESULTS: The [Ca(2+)]i in LEC was obviously increased in the untrated group as compared with that in the normal control group (P<0.01), and the [Ca(2+)](i) in LEC in the curcumin-treated group was highest among three groups (P<0.01). The content of intracellular cAMP in LEC was decreased while the content of intracellular cGMP was obviously increased in the untreated group as compared with those in the normal control group (P<0.01). The content of intracellular cAMP in LEC was higher in the curcumin-treated group than that in the untreated group, while the content of intracellular cGMP was lower than that in the untreated group (P<0.01). CONCLUSION: The antiproliferation effects of curcumin on LEC may relate to the regulations of multiple processes of signal transduction.