OBJECTIVE:To prepare silybin liposomes(SLBL)coated by N-trimethyl chitosan(TMC)(TMC60-SLBL),and to optimize the formula. METHODS:The effects of 3 kinds of preparation methods on encapsulation efficiency of SLBL were com-pared,including film dispersion method,multiple emulsion method and reverse evaporation method. The formula of TMC60-SLBL was optimized by orthogonal test using encapsulation efficiency as index with the concentration of phospholipid,ratio of phospholip-id to cholesterol,ratio of silybin to lipids,hydration temperature as factors. The stability of TMC60-SLBL by optimal formula at 4 and 25℃within 30 d,the particle size and Zeta potential of TMC60-SLBL and SLBL were all compared. RESULTS:The encapsu-lation efficiency of TMC60-SLBL prepared by the film dispersion method was the highest,without statistical significance in encap-sulation efficiency before and after cutting (P>0.05). The optimal formula was with the concentration of phospholipid 6 mg/ml;the ratio of phospholipid to cholesterol 40∶1;the ratio of silybin to lipids 1∶30;the temperature of hydration medium 45 ℃;the encapsulation efficiency were(82.08±2.6)%,RSD=3.17%(n=3). The preparation was stable at 4℃;the mean diameter of SL-BL and TMC60-SLBL were(131.9±1.9)nm and(161.2±2.0)nm,and Zeta potential respectively were(-23.18±1.14)mV and (36.73 ± 2.84) mV. CONCLUSIONS:TMC60-SLBL is prepared successfully,and its formula is simple and practicable with high encapsulation efficiency.

OBJECTIVETo evaluate the early clinical effects of arthroscopic treatment for cruciate ligament cysts of the knee.

METHODSFrom September 2008 to January 2014, 23 patients with cruciate cysts were treated with arthroscopic surgery. There were 11 males and 17 females, with an average age of 28.3 years old (ranged, 16 to 56 years old). Twenty patients had injuries on the right knee and 8 patients had injuries on the left knee. Eight patients had a history of injury,5 patients had a history of chronic injury,and the other 15 patients had no obvious reasons for the symptom. Before the operation, 24 patients were clearly diagnosed as the cruciate ligament cysts of knee joint,and 4 patients were diagnosed as other problems of the knee, but the diagnosis were corrected after the operation. According to the MRI before the surgery,all the patients could be divided into 3 types: 14 were type I, 6 were type II, 8 were type III. After the operation, the patients were suggested to have a rest for 2 weeks, and take exercises everyday at the same time. The knee range of motion, Lysholm score, International Knee Documentation Committee (IKDC) 2000 subjective score, GLASGOW criteria, and McMurray test, pivot shift test, and anterior drawer test were observed to evaluate clinical results.

RESULTSAll the incisions healed at the first stage without complications. Twenty-five patients were followed up, and the duration ranged from 12 to 52 months, with a mean of 26.7 months. Three patients accepted the reconstruction of ACL or tightened elongated ACL using bipolar radio frequency. The Lysholm score increased from preoperative 59.80 +/- 6.58 to 75.32 +/- 6.49 at the latest follow-up; IKDC 2000 score increased from preoperative 65.36 +/- 6.26 to 81.00 +/- 5.76 at the latest follow-up. According to GLASGOW criteria,23 patients got an excellent result and 2 good.

CONCLUSIONIt has a satisfactory curative effect on ACL reconstruction using bipolar radio frequency. It has advantages of firm fixation, simple and secure operation as well as quick postoperative recovery. It's very important to check the MRI before operations, and carefully seek the cysts during the operations, avoiding the omission of any cysts,especially the cases of multiple cysts. Finally, patients should take exercises actively after operations.

Adolescent ; Adult ; Anterior Cruciate Ligament ; physiopathology ; surgery ; Anterior Cruciate Ligament Injuries ; Anterior Cruciate Ligament Reconstruction ; Arthroscopy ; Cysts ; surgery ; Female ; Humans ; Knee Injuries ; physiopathology ; surgery ; Knee Joint ; physiopathology ; surgery ; Male ; Middle Aged ; Range of Motion, Articular ; Young Adult

OBJECTIVETo investigate the mechanism of liraglutide-mediated protection against nonalcoholic fatty liver disease (NAFLD) using aApoE knockout (KO) mouse with high-fat diet (HFD) and Acrp30 knockdown.

METHODSFifty-six male ApoE KO mice were divided into the following six modeling and experimental groups:regular chow fed (ApoE KO, n=10), HFD fed (HF, n=10), HFD+Adenovirus (Ad)-small hairpin (sh) Acrp30 (Ad-shAcrp30, n=10), HFD+Ad-shGreen Fluorescent Protein (GFP) (Ad-shGFP, n=6), HFD+Ad-shAcrp30+liraglutide (liraglutide, n=10), and HFD+Ad-shAcrp30+saline (saline, n=10). Weight-matched C57BL/6 mice on the regular chow diet were used as the control group (WT control, n=10).All mice were fed their assigned diet for 16 weeks.The Ad-shGFP or Ad-shAcrp30 was injected by tail vein at the end of 14 and 15 weeks.Mice in the liraglutide group received 1 mg/kg of the drug, twice daily, intraperitoneally for a total of 8 weeks (from the 9th to 16th week).Fasting blood samples were collected for testing levels of fasting plasma glucose (FPG), triglycerides (TGs), total cholesterol (TC), free fatty acid (FFA), alanine aminotransferase (ALT), Acrp30 and insulin.Liver tissue was procured for histological examination.Expression of mRNA was detected by real-time RT-PC and of protein was detected by western blot analysis.

RESULTSThe Ad-shAcrp30 treated mice had reduced expression of Acrp30 at both the mRNA and protein levels in adipose tissues and plasma, as compared with the AdshGFP treated mice (all P < 0.01).Compared to the WT and ApoE KO groups, the HF group showed higher levels of FPG, FFA, TGs and TC (all P < 0.01); furthermore, the Ad-shAcrp30 treatment compounded these changes.The Ad-shAcrp30 treated group had markedly higher hepatic TC and TGs than the HF group (P < 0.01 and P < 0.05).Oil Red O staining showed that there was more lipid droplets in the liver tissue of the Ad-shAcrp30 treated group than in that of the HF group (P < 0.01), and hematoxylin-eosin staining confirmed these results.Liraglutide treatment prevented the increase in body weight, FPG, FFA, TGs, TC and ALT levels, as compared to the saline controls (all P < 0.01), but the plasma Acrp30 levels and the Acrp30 mRNA and protein expression in adipose tissues were elevated (all P < 0.01).Oil-Red O staining indicated that the liraglutide group had a significantly lower hepatic lipid content than the saline group, and total hepatic TG and TC were reduced in the former group (P < 0.01 and P < 0.05).The liraglutide treatment significantly attenuated the mRNA expression of ACC and FAS (both P < 0.01) but increased AMPK phosphorylation (P < 0.01).

CONCLUSIONAdministration of liraglutide prevented the development of HFD-and hypoadiponectinemia-induced metabolic disturbance and accumulation of hepatic lipids in this mouse model system of NAFLD.

AMP-Activated Protein Kinases ; metabolism ; Adiponectin ; deficiency ; metabolism ; Adipose Tissue ; Alanine Transaminase ; Animals ; Apolipoproteins E ; deficiency ; metabolism ; Cholesterol ; Diet, High-Fat ; Disease Models, Animal ; Glucagon-Like Peptide 1 ; analogs & derivatives ; Insulin ; Liraglutide ; Male ; Metabolism, Inborn Errors ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Non-alcoholic Fatty Liver Disease ; metabolism ; Protective Agents ; RNA, Messenger ; Triglycerides

OBJECTIVETo establish the methods of calculating and analyzing the multi-coefficient of variation significance test for the toxicology study.

METHODSThe paper aimed to confirm the significance level with the method of Bonferroni and then compared the methods of calculating and analyzing of the experiment groups with the control group respectively.

RESULTSThe significance level of multi-coefficient of variation significance test was confirmed as alpha1=0.0167. Compared with the control groups, the activity of ALT in serum both in 30 mg/kg and 60 mg/kg groups did not change in the average significance test, which was not statistically significant (P>0.05), while it changed in the variation significance test, which was of statistical significance (P<0.0167). The activity of AST in serum in 60 mg/kg group did not change in the average significance test (P>0.05), while it changed in the variation significance test (P<0.0167).

CONCLUSIONThe complete changes of the indexes can only be shown by use of both the average significance test and the variation significance test together.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Disease Models, Animal ; Female ; Lead Poisoning ; enzymology ; Rats ; Rats, Wistar ; Statistical Distributions

To investigate the non-viral vector mediating human coagulation factor VIII gene expression in mouse 32D cell line, a recombinant plasmid vector, pRC/RSV-hFVIIIBDcDNA, was constructed by cloning B-domain-deleted (Delta760aa-1639aa) human factor VIII cDNA (hFVIIIBDcDNA) into plasmid vector, pRC/RSV. The plasmid RC/RSV-hFVIIIBDcDNA was then transfected by means of SuperFect Transfection Reagent into mouse 32D cell line. After screening with G418, the procoagulant activity (hFVIII:C) and antigen (hFVIII:Ag) of human factor VIII in the culture medium were detected using one-stage method and ELISA, respectively. Furthermore, RT-PCR was performed to observe the transcription of hFVIIIBDcDNA. The results showed that human coagulation factor VIII protein existed in culture medium with hFVIII:C up to 2.01 U/(10(6) cell x 24 hours) and hFVIII:Ag to 450.08 ng/(10(6) cell x 24 hours). RT-PCR displayed mRNA of hFVIIIBDcDNA in 32D cells. It is concluded that the recombinant plasmid RC/RSV-hFVIIIBDcDNA can successfully express human FVIII in mouse 32D cell line, and hFVIII expressed in vitro presents the similar coagulant activity to the native hFVIII existing in normal human plasma.
Animals ; Cell Line ; DNA, Complementary ; genetics ; Enzyme-Linked Immunosorbent Assay ; Factor VIII ; genetics ; metabolism ; Gene Expression Regulation ; Humans ; Mice ; Plasmids ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo observe the protective effect of Panax notoginseng saponins (PNS) on the level of synaptophysin ptotein in brain in rat model with Alzheimer's disease (AD).

METHODThe AD rat models were established by intra-peritoneal injection of D-galactose combined with excitatory neurotoxin ibotenic acid injection into bilateral nbM. The activity and content of synaptophysin protein in brain were determined by immunohistochemistry analysis.

RESULTPNS could reduce the lesion of level of synaptophysin protein in brain, as compared with those of model group's rats.

CONCLUSIONPNS plays a protective role by reducing down of the level of synaptophysin protein in brain in lesion of AD animal model.

Alzheimer Disease ; chemically induced ; metabolism ; pathology ; Animals ; Basal Nucleus of Meynert ; drug effects ; pathology ; Brain ; metabolism ; pathology ; Galactose ; toxicity ; Ginsenosides ; isolation & purification ; pharmacology ; Ibotenic Acid ; toxicity ; Neuroprotective Agents ; isolation & purification ; pharmacology ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Synaptophysin ; metabolism

OBJECTIVETo study the expression of a glycoprotein of plant origin in normal, benign and malignant breast tissues.

METHODSExpression of a plant glycoprotein was examined in 5 samples of normal breast tissues, 20 fibro-adenoma and 136 breast cancer by SABC immunohistochemical staining and the results were analyzed by SPSS statistics software.

RESULTSNo positive staining was detected in normal breast tissues (0/5). Weak staining was observed in 4 of 20 (20.0%) breast fibro-adenoma. Positive staining was demonstrated in 116 out of 136 (85.3%) breast cancer specimens. The differences were statistically significant. The expression of plant-associated human cancer antigen was related to pathological grade (P < 0.05), tissue invasiveness (P < 0.01) and recurrence (P < 0.05), but not to patients' age, tumor size and c-erbB-2 expression.

CONCLUSIONThe plant glycoprotein studied may be a human cancer-associated antigen which might be a potential marker of breast cancer.

Adenocarcinoma, Mucinous ; immunology ; pathology ; Adult ; Aged ; Antigens, Neoplasm ; metabolism ; Biomarkers, Tumor ; metabolism ; Breast ; immunology ; Breast Neoplasms ; immunology ; pathology ; Carcinoma, Ductal, Breast ; immunology ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; immunology ; pathology ; Female ; Fibroadenoma ; immunology ; pathology ; Humans ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; metabolism ; Plants ; immunology ; Receptor, ErbB-2 ; metabolism

BACKGROUNDAnomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) is a rare congenital heart anomaly. We aimed to illustrate the clinical features and long-term prognosis of patients with ALCAPA.

METHODSTwenty three patients (13 males and 10 females, ages ranging from 2.5 months to 65 years) identified as ALCAPA in Beijing Anzhen Hospital from April 1984 to June 2009 were divided into two groups, based on the age of onset: group 1 (≤ 12 months, n = 16) and group 2 (> 12 months, n = 7).

RESULTSFifty six point three percent of patients in group 1 had been misdiagnosed as endocardial fibroelastosis (9/16), 18.8% as dilated cardiomyopathy (3/16) and 6.3% as myocardial infarction (1/16). Patients in group 2 were usually diagnosed as coronary heart disease, myocarditis, or patent ductus arteriosus. Electrocardiography in group 1 revealed abnormal Q waves with T wave inversion in leads I, avL, V(4)-V(6), especially in lead avL (deep and wide Q wave); but no specific manifestations in group 2. A higher percentage of patients in group 1 had cardiomegaly on chest radiograph (86.7% vs. 33.3%, P = 0.031), while pulmonary artery protrusion was more common in group 2 (26.7% vs. 83.3%, P = 0.046). Lower left ventricular ejection fraction (LVEF) was present in group 1 than in group 2 ((48.5 ± 11.5)% vs. (65.0 ± 6.1)%, P < 0.001). Apical ventricular aneurysm (62.5% vs. 0%, P = 0.007), enhanced echogenicity of papillary muscles (87.5% vs. 28.6%, P = 0.011) and endocardial thickening (93.8% vs. 14.3%, P < 0.001) were more frequent in group 1 than in group 2. The ratio of the proximal right coronary artery (RCA) diameter to the aortic root diameter exceeded 0.14 in all cases, more prominent in group 2 (0.26 ± 0.05 vs. 0.33 ± 0.03, P = 0.009). Increased coronary artery collaterals within the interventricular septum were detected in 18 patients (78.3%) by Doppler imaging. Twenty one patients underwent cardiac surgery, including left coronary artery (LCA) ligation (1/21), LCA ligation plus coronary artery bypass grafting (1/21), Takeuchi operation (7/21), and LCA reimplantation surgery (12/21). Four patients underwent concomitant mitral valve repair and one received mitral valve replacement. Aneurysm resection was performed in 3 cases. Six patients died in hospital after surgery, and the rest of the cohort had no overt symptoms during a follow-up period of 6 to 166 months. Their abnormal Q waves gradually regressed or disappeared, and the LVEF and left ventricle size returned to normal range with alleviation of mitral insufficiency.

CONCLUSIONSThe accurate diagnosis of ALCAPA can be made with serial diagnostic methods. ALCAPA can be successfully treated with several types of cardiac surgery, and surgeries of establishing two-coronary-artery circulation are the preferred operations nowadays, with good long-term prognosis.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Coronary Vessel Anomalies ; diagnosis ; mortality ; Echocardiography ; Electrocardiography ; Female ; Follow-Up Studies ; Humans ; Infant ; Male ; Middle Aged ; Prognosis ; Pulmonary Artery ; abnormalities

OBJECTIVETo observe the ECG and electrophysiological characteristic of patients with idiopathic ventricular tachycardia (VT) and premature ventricular contraction (PVC) originating from left (LVOT) and right (RVOT) ventricular outflow tracts and assess the clinical effect of radio frequency catheter ablation (RFCA) on these patients.

METHODSRFCA was performed in 58 patients (10 with VT and 48 with PVC, 5 patients with VT from RVOT under the guidance of non-contact mapping system Ensite3000). VT or PVC originated from LVOT in 15 patients (12 out of 15 from left sinus of Valsalva) and RVOT in 43 patients.

RESULTS(1) R wave in II, III, aVF leads was the common characteristics of VT or PVC originated from LVOT and RVOT and difference in wave duration index and R/S-wave amplitude ratio in V(1) or V(2) could be used to define VT and PVC originated from LVOT or RVOT. (2) Ablation was successful in 55 out of 58 patients (9 patients with the 2nd ablation, evaluated as arrhythmia-free at 3 months post ablation without medication) and failed in 3 patients. One patient developed pericardial tamponade during ablation and recovered without complication after related treatments.

CONCLUSIONSRFCA is an effective, safe and curative therapy for VT or PVC originated from LVOT and RVOT. Non-contact mapping system (Ensite3000) is a safe and reliable tool to guide mapping and ablation in patients with complex VT and unstable hemodynamics.

Adolescent ; Adult ; Aged ; Catheter Ablation ; Female ; Humans ; Male ; Middle Aged ; Tachycardia, Ventricular ; etiology ; therapy ; Ventricular Outflow Obstruction ; complications ; Ventricular Premature Complexes ; etiology ; therapy ; Young Adult

Aim To assess the value of the administration of adenosine-5'-triphosphate (ATP) during sinus rhythm for noninvasive diagnosis of AV node dual pathways(AVNDP) and abolition or modification of the slow pathway (SP) after radiofrequency(RFCA) in patients with inducible sustained AVNRT. Methods Incremental doses of ATP were intravenously administrated during sinus rhythm to patients with spontaneous or inducible sustained AVNRT(study group, n=45)and to patients with no evidence of AVNDP or inducible AVNRT (control group, n=37) until ECG signs of AVNDP( 50 ms increase or decrease in P-R interval in two consecutive beats, or occurrence of AVNRT) or second-degree AV block were observed. Results Four patients (two in study patients and two in control patients) could not complete the trial and were excluded from analysis. AVNDP was observed by ATP in 36(84%) study patients, whereas it was diagnosed by electrophysiology criteria in 38(88%) patients. AVNDP was observed only in 1(3%) control patient. AVNDP by ATP test was disappeared in 18(90%) of 20 patients who underwent SP abolition and in 3(38%) of 8 patients who underwent SP modification. Conclusion ATP test during sinus rhythm enables noninvasive diagnosis of AVNDP in a high percentage of patients with inducible AVNRT and reliably confirms the results of RFCA of the SP.