Objective To investigate the retainage rate,distribution,and emigration of the bone marrow mesenchymal stem cell(BMMSC) after transcoronary infusion and to further evaluate the feasibility of injecting BMMSCs into coronary artery.Methods BMMSCs were isolated,purified,expanded,and labelled with CM-DiI.The infarcted SD rat hearts were removed and perfused with Langendorff apparatus.The cells were injected into the aortic root and the fluid returning from the coronary system was collected and the labelled cells in the coronary effluent were quantitated with flowcytometry.At the same time,left ventricle function was recorded to evaluate the safety of this approach.In vivo study,the cells were then injected into clamped ascending aorta through a catheter inserted through the left ventricle into the aortic root.The hearts were harvested at different time points after cell transplantation to obtain the direct evidence of distribution and emigration of the implanted cells.Results Only 3%～5% of transplanted cells returned into the right ventricle and more than 90% cells retained in the heart after beinginjected into aortic root of Langendorff model of infarcted hearts of SD rats.Left ventricle function did not deteriorate after cell transplantation.The labelled cells were entrapped within the coronary capillary immediately after cell infusion,mainly in the normal area.After 24 hours some cells migrated through the capillary wall into interstitium of the heart.One week later we found that most survival cells located at the infarcted area and the border zone.Conclusion The majority of BMMSCs delivered by transcoronary infusion retained in the heart.BMMSCs can penetrate the vessel wall and home back to the interstitial compartment and the injured area in a few hours.

OBJECTIVETo investigate the metabolism-based interaction between ipriflavone(IP) and other drugs in vitro.

METHODSIP and propafenone or estradiol, or other drugs were co-incubated with rat liver microsome induced by beta-naphthoflavone. The concentrations of residual substrate in microsomal incubates were determined by HPLC.

RESULTThe metabolism of propafenone (10 microg/ml) incubated with 2 microg/ml or 50 microg/ml IP was markedly inhibited compared with control group(P<0.01). While the metabolism of 10 microg/ml of estradiol incubated with 10 microg/ml and 100 microg/ml IP showed no marked inhibition. When IP (20 microg/ml) was incubated with 0.5 microg/ml propranolol, 0.5 microg/ml propafenone or 5.0 microg/ml estradiol respectively the metabolic activity of IP was markedly inhibited compared with control group (P<0.05 P<0.05 P<0.02 respectively).

CONCLUSIONThe results indicate that there is an interaction between ipriflavone and propafenone during in vitro metabolism, and there might also be clinical significance.

Animals ; Drug Interactions ; Estradiol ; metabolism ; Female ; Isoflavones ; metabolism ; Propafenone ; metabolism ; Propranolol ; metabolism ; Rats ; Rats, Sprague-Dawley

AIMTo establish a RP-HPLC method for determination of diphenytriazol (DL111-IT) in rat hepatic microsomes.

METHODSDL111-IT in rat hepatic microsomal incubates was extracted with chloroform, using diazepam as internal standard. The determination was performed on a Lichrospher ODS-C18 reversed column (25 cm x 0.46 cm ID) with mobile phase of methanol-pH 7.5 phosphate buffer (70:30) at a flow-rate of 1.0 mL.min-1. A UVVIS detector was operated at 235 nm.

RESULTSThe assaywas linear from 1.01-101.0 micrograms.mL-1 for DL111-IT. The limit of detection was 0.15 microgram.mL-1 (signal-to-noise ratio 3) and the limit of quantification was 1.01 micrograms.mL-1(RSD < 10%, n = 4). The method afforded average recoveries of (100.3 +/- 1.9)% (n = 5), and intra-day and inter-day RSD were less than 5.0%(n = 5). The method allowed study of the in vitro phase I metabolism of DL111-IT in rat liver microsomal incubates. The microsomes induced by beta-naphthoflavone showed high enzymatic activity for DL111-IT phase I metabolism.

CONCLUSIONThe method is simple, accurate and can be used to study the metabolism of DL111-IT in rat hepatic microsomes.

Abortifacient Agents, Nonsteroidal ; analysis ; metabolism ; Animals ; Cell Separation ; Chromatography, High Pressure Liquid ; methods ; Female ; Microsomes, Liver ; metabolism ; Rats ; Rats, Sprague-Dawley ; Triazoles ; analysis ; metabolism

BACKGROUND: Currently, most of the clinical trials of cell transplantation for ischemic heart disease is the transplantation of bone marrow mononuclear cells through the bypass graft artery in patients with acute myocardial infarction, but reports in combination with cell transplantation for old myocardial infarction are few. OBJECTIVE: To investigate the safety and feasibility of intracoronary artery injection of bone marrow mononuclear cells through the bypass graft artery during coronary artery bypass grafting (CABG). DESIGN: Self-control and case analysis. PARTICIPANTS: A total of 10 patients who had old myocardial infarction, left ventricular ejection fraction (LVEF)≤40%, were selected from Cardiovascular Institute and Fuwai Hospital from November 2004 to June 2005. METHODS: The bone marrow mononuclear cells were harvested from the bone marrow by Ficoll density gradient centrifugation method before the CABG was carried. And the patients received CABG and 10 mL mononuclear cell suspension through the grafts into anterior descending branch. In addition, 10 mL mononuclear cell suspension was injected into the circumflex branch and right coronary artery through the proximal heart. MAIN OUTCOME MEASURES: The heart function was evaluated with transthoracic echocardiography (TEE) and cardiac MRI after the operation. RESULTS: All patients recovered. A total of 45-60 mL bone marrow was harvested from iliac crest, and 4.1 ×10~7 mononuclear cells were isolated and identified by trypan blue test (cell activity >95%). TEE showed that the LVEF at 1 week and 1, 3 months postoperatively was significantly improved compared with before operation; creatase arid troponin T were not increased, and no myocardial infarction changes were found. MRI showed that the LVEF was significantly increased following operation (P < 0.01); left ventricular end-diastolic and systolic diameters were significantly decreased (P < 0.05). There was no complication associating with bone marrow harvest, or cell transplantation. CONCLUSION: Bone marrow mononuclear cells transplantation through bypass graft, as an adjunctive therapy, is safe and feasible.

OBJECTIVETo develop an analytical method and quality control for determination of zolmitriptan and related substances.

METHODSZolmitriptan and related substances were separated and determined on a shimadzu CLC-C(8) column (150 mm x 6 mm, 10 microm) with a mobile phase of acetonitrile-10 mmol/L phosphate buffer (25:75 pH 7.5) and a flow-rate of 1 ml/min; the UV-VIS detector was operated at 229 nm.

RESULTThe limit of detection for the related substances was 0.5 ng on the zolmitriptan basis (S/N >3). Linear calibration curve was gene rated from 4 - 40 microg/ml with a correlation coefficient of 0.9999. The recovery rate of zolmitriptan was 99.1% with a standard deviation of 0.2%. The results of HPLC method were consistent with those of nonaqueous titration method.

CONCLUSIONHPLC method is a rapid sensitive and accurate method for the determination of zolmitriptan and its related substances.

Chromatography, High Pressure Liquid ; Oxazolidinones ; analysis ; Tryptamines

Objective To evaluate the therapeutic efficacy and safety of transcatheter closure of atrial septal defect (ASD) and patent ductus arteriosus (PDA) using Amplatzer occluder. Methods Routine cardiac catheterization and angiography were performed in 50 patients (23 male, 27 female, age ranging from 3 to 64 years old), including 19 cases of ASD and 31 cases of PDA under local or general anesthesia. After balloon sizing of the ASD, the optimal Amplazter septal occluder (ASO) was transmitted into the left atrial, and the left and right disks were released in turn. The Amplatzer occluder was completely released after transthoracic echocardiography confirmed that there was no residual shunts or new onset mitral valve regurgitation. The Amplatzer duct occluder (ADO) size was selected according to the narrowest point of PDA measured by angiography, and the occluder was released after the repeated angiography showed no residual shunts. Results ① The mean diameter of the ASD measured by balloon was 13-31 (23?6) mm and the diameter of ASO was (17-40) mm. The immediate closure rate was 100%. ② Angiography confirmed that closure of the ductus using ADO was achieved in 30 patients, and closure of the large size (12 mm) was achieved in 1 case of PDA patient using ASO (17 mm). No complications were encountered. Conclusion Transcatheter closure of ASD and PDA using Amplatzer device, with the advantages of simple operation, confirmative occlusion efficacy, minimal invasiveness, wide indications, and less complications, has a bright future of clinical application.

Objective To evaluate the two-stage repair in hypospadias cripples.Methods 35 patients with hypospadias cripples underwent two-stage repair.At the first stage,a full-thickness graft of skin or buccal mucosa was used for urethral plate reconstruction after the release of chordee.The second stage was performed 6 months after completion of the first stage.Tubularization of the replaced urethral plate was the preferred technique.Results There was no urethra stricture or “preplaced urethral plate ” atrophy.Fistula developed in 2 of 35 cases.Conclusion The twostaged technique was a useful strategy to deal with the myriad abnormalities encountered in crippled hypospadias.This technique could not only create a neourethra successfully,but also give the penis a near-normal shape and appearance.

AIMTo observe the metabolic interaction between diphenytriazol and steroid hormone drugs, and provide some useful information for clinical medication.

METHODSThe steroid hormone drugs which may be co-administrated with diphenytriazol were selected, such as mifepriston, estradiol, medroxyprogesterone acetate, progesterone, norethisterone and so on. Diphenytriazol was incubated with each drug in rat liver microsome. The residual concentration of diphenytriazol or steroid hormone drugs in the microsomal incubates was determined by reversed-phase high-performance liquid chromatography, separately. The inhibition constants (K(i)) for each of them were calculated.

RESULTSThe inhibition constant K(is) of diphenytriazol for the metabolism of mifepristone, estradiol, medroxyprogesterone acetate, progesterone and norethisterone were (201.3 +/- 1.0), (94 +/- 4), (128.7 +/- 2.2), (64 +/- 5) and (80 +/- 4) micromol x L(-1), respectively. The inhibition constants K(i) of steroid hormone drugs for the metabolism of diphenytriazol was (66.9 +/- 2.2) micromol x L(-1) for estradiol, (60.0 +/- 2.3) micromol x L(-1) for medroxyprogesterone acetate, (163 +/- 10) micromol x L(-1) for progesterone and (88 +/- 5) micromol x L(-1) for norethisterone, respectively.

CONCLUSIONDiphenytriazol shows metabolism interaction with steroid hormone drugs such as estradiol, medroxyprogesterone acetate, progesterone and norethisterone.

Abortifacient Agents, Nonsteroidal ; metabolism ; pharmacology ; Abortifacient Agents, Steroidal ; metabolism ; Animals ; Contraceptives, Oral, Synthetic ; metabolism ; Drug Interactions ; Estradiol ; metabolism ; Female ; In Vitro Techniques ; Medroxyprogesterone ; metabolism ; Microsomes, Liver ; metabolism ; Mifepristone ; metabolism ; Rats ; Rats, Sprague-Dawley ; Triazoles ; metabolism ; pharmacology

Objectives To access the bacteriology in patients with sepsis due to biliary tract infection to provide a basis for empirical selection of proper antibiotic treatment.Methods This is a single-center retrospective study on 214 patients with biliary tract infection admitted from August 2014 to July 2016 to the surgical intensive care units (ICU) of The First Affiliated Hospital of Sun Yat-sen University.To study the demographic information,sequential organ failure assessment (SOFA),usage of antibiotics before ICU and duration of ICU were analyzed.Bile,peritoneal drainage and blood samples were collected.Results 47 septic shock patients and 25 septic patients due to biliary tract infection were enrolled in the trial.The two groups (the shock group vs.the sepsis group) had a significant difference in the duration of ICU stay [(6.4 ± 4.6) d vs.(2.3 ± 1.8) d,P ＜ 0.05].48 strains of pathogens were isolated from the bile samples.The major pathogens were Escherichia coli (E.coli) (n =23,47.9％),Enterococcus faecalis (n =8,16.7％) and Enterococcus faecium (n =2,4.2％).80 strains of pathogens were isolated from the peritoneal drainage culture samples.E.coli,pseudomonas aeruginosa,and Klebsiella pneumoniae ranked the top 3 species,accounting for 26.3％,11.3％ and 7.5％,respectively.The sensitivity of E.coli isolated from bile to amikacin,imipenem and panipenem were all over 90.0％.Conclusions E.coli was the principal gram-negative bacterium in biliary infection induced sepsis.Early administration of carbapenemes may reduce the occurrence of septic shock in these patients.

Skeletal muscle is the largest tissue in the human body, and it plays a major role in exerting force and maintaining metabolism homeostasis. The role of muscle transcription factors in the regulation of metabolism is not fully understood. MondoA is a glucose-sensing transcription factor that is highly expressed in skeletal muscle. Previous studies suggest that MondoA can influence systemic metabolism homeostasis. However, the function of MondoA in the skeletal muscle remains unclear.

We generated muscle-specific MondoA knockout (MAKO) mice and analyzed the skeletal muscle morphology and glycogen content. Along with skeletal muscle from MAKO mice, C2C12 myocytes transfected with small interfering RNA against MondoA were also used to investigate the role and potential mechanism of MondoA in the development and glycogen metabolism of skeletal muscle.

MAKO caused muscle fiber atrophy, reduced the proportion of type II fibers compared to type I fibers, and increased the muscle glycogen level. MondoA knockdown inhibited myoblast proliferation, migration, and differentiation by inhibiting the phosphatase and tensin homolog (PTEN)/phosphoinositide 3-kinase (PI3K)/Akt pathway. Further mechanistic experiments revealed that the increased muscle glycogen in MAKO mice was caused by thioredoxin-interacting protein (TXNIP) downregulation, which led to upregulation of glucose transporter 4 (GLUT4), potentially increasing glucose uptake.

MondoA appears to mediate mouse myofiber development, and MondoA decreases the muscle glycogen level. The findings indicate the potential function of MondoA in skeletal muscle, linking the glucose-related transcription factor to myogenesis and skeletal myofiber glycogen metabolism.