Objective:To establish an HPLC method for the determination of the content of magnolol and honokiol in Cortex mag-noliae Officinalis formula granules and compare the content of the formula granules from different manufacturers. Methods:An HPLC was used to determine the content of magnolol and honokiol in Cortex magnoliae Officinalis formula granules. The analysis was carried out on a Hypersil C18 (250 mm × 4. 6 mm, 5 μm) chromatographic column. Acetonitrile-water was used as the mobile phase with gra-dient elution and the flow rate was 1. 0 ml·min-1 . The detection wavelength was set at 294 nm, the sample size was 20 μl and the column temperature was 25℃. Results:The linear range was 0. 873-26. 190μg·ml-1(r=0. 999 5) for magnolol, the average recov-ery was 99. 24% with RSD of 2. 00%(n=6) and that was 0. 732-21. 980μg·ml-1(r=0. 999 0) for honokiol,and the average recov-ery was 99. 89% with RSD of 1. 33%(n=6). The difference in the content of magnolol and honokiol in Cortex magnoliae Officinalis formula granules from different manufacturers was notable. Conclusion: The method is simple, repeatable and feasible, and can be used for the quality control of magnolol and honokiol in Cortex magnoliae Officinalis formula granules. The content difference in magno-lol and honokiol in Cortex magnoliae Officinalis formula granules from different manufacturers suggests that it is necessary to standardize the planting and selecting of Chinese medicine, and develop scientific and unified production technology and quality standard for the formula granules.

A reporter system for φC31 integrase was developed in NIH3T3 cells. The reporter plasmid coding green fluorescent protein (GFP) coupled with red fluorescent protein (RFP) was eo-transfected with the plasmid coding φC31 integrase, to show the activity of integrase in the cells. Fluorescence activated cell sorter (FACS) was used to measure the proportion of the cells containing red and green fluorescence. The increment of green cells was positively related to the increase in the transfection with plasmid coding φC31 integrase. Approximately 90% of green cells were observed under a ratio of [plasmid-φC31-integrase]/[reporter plasmid] at 10 : 1. This suggests that the φC31 integrase reporter system provides a probe for the function of φC31 integrase in cells.

A reporter system for ?C31 integrase was developed in NIH3T3 cells.The reporter plasmid coding green fluorescent protein(GFP) coupled with red fluorescent protein(RFP) was co-transfected with the plasmid coding ?C31 integrase, to show the activity of integrase in the cells.Fluorescence activated cell sorter(FACS) was used to measure the proportion of the cells containing red and green fluorescence.The increment of green cells was positively related to the increase in the transfection with plasmid coding ?C31 integrase.Approximately 90% of green cells were observed under a ratio of plasmid-?C31-integrase/reporter plasmid at 10∶1.This suggests that the ?C31 integrase reporter system provides a probe for the function of ?C31 integrase in cells.

Objective To investigate the effects of intravenous infusion of lidocaine on colorectal resection.Methods Sixty patients undergoing colorectsl resection were randomly divided into group L [receiving intravenous infusion of 1%lidocaine,bolus injection of 1.5 mg/kg lldocaine at induction of anesthesia,followed by a continuous infusion of 1.5 mg/(kg·h)intraoperatively and for 12 h postoperatively]and group C(an equal volume of saline).Postoperative pain scores(VAS score)and morphine consumption were measured.Times of flatus and first defecation were recorded.Results The end expiration sevotturane concentration maintaining hemedynamics stable in group L was lower than that in group C(P<0.05).The dose of sulfentsnyl was(17.5±3.8)μg in group L,(25.6±4.5)μg in group C,there was significant difference between two groups(P<0.05).The times of flatus first,defecation and morphine conaumption at 24 and 48 h after operation in group L were lower than those in group C[(23±6)h vs(28±7)h,(31±7)h vs(43±9)h,(32±6)mg vs(46±8)nag and(58±7)mg vs(71±10)mg,P<0.05].There was no significant difference at resting VAS score,but there was significant difference at movement and cough VAS score between twp groups(P<0.05).Conclusion Perioperative administration of low doses of intravenous infusion of lidocaine reduces intraoperative anesthetic requirements and has a clinically relevant beneficial effect on postoperative recovery after colorectal resection.

Objective To establish the methodology of flow cytometry for detecting human cells in human/goat chimerisra.Methods Human hemopoietic stem/progenitor cells (CD+34 cells) or MIG-tranadueed-GFP CD+34 cells were transplanted into the peritoneal cavity of fetal goats in utero to obtain human/goat chimera modeL The peripheral blood cells from the chimeras were labeled with multiple mouse anti-human antibodies and the monoelonal antibodies that were specific for human but had not or only minimal cross-reaction with goat were screened as the primary antibodies for routine analysis in flow cytometry.Human cord blood was proportionally (25% ,50% ,75%,100%) added into the blood of the untransplanted goats and the cells were labeled with CD+34 monoclonal antibody.The region and size of the "gate" were chosen based on to the distribution of CD+34 cells or human cord blood.One human/goat chimera marked with GFP (MIG goat) was sacrificed and the substantial liver cells from its perfused liver were analyzed for the GFP+cells percentage and DNA contents by flow cytometry.Results CD7,CD15,CD38,CD45CD20CD34CD14and GPA monoclonal antibodies were chosen as the primary antibodies in rou tine detection by flow cytometry.The size and area of the "gate" were also defined.29.1% (29100/100 000 ) of the substantial liver cells from the MIG goat expressed GFP.DNA content analysis showed that the GFP+ cells obtained from the liver of MIG goat mainly manifested two peaks that were correspond to those of human.Conclusions Flow cytometry is rapid,simple and effective for the investigation of differentiation,homing and biological characteristics of stem cells in vivo.The selections of suitable surface antibodies and the "gate" are very important for detecting human cells accurately in the human/goat chimerism.

Objective To evaluate the effect of greater trochanteric transfer for the treatment of coxa brevis in children and adolescents.Methods From August 2005 to January 2011,twenty patients (22 hips)with coxa brevis underwent greater trochanteric transfer.Among them 18 patients (20 hips) were available for evaluation,including 4 boys and 14 girls,with an average age of 11.4 years (range,7.5-15.0 years) at operation.Five cases (6 hips) were caused by Perthes disease,and 13 cases (14 hips) were caused by developmental dysplasia of hip.Four patients (4 hips) had undergone greater trochanteric epiphyseodesis ever before.All patients were fixed with tension screw after the deformity was corrected during the operation.After operation,the human plaster spica was used for 6 weeks in all patients.Results All patients were followed up for 14 to 79 months (average,31 months).At the last follow-up,fatigue or pain in the hips disappeared or improved in 13 patients.Sixteen patients had limping and positive Trendelenburg sign preoperatively,at the last follow-up 9 patients got improvement.Twelve patients (13 hips) had limitation of abduction of the hip,the average range of abduction was 25.38°±1.20°,which was improved to 45.38°±1.05° at the last follow-up.The average articulotrochanteric distance and ratio of the distance from the greater trochanter tip to femoral head center and the radius of the femoral head at the last follow-up was (17.47+3.14)mm and 2.10±0.21,respectively,there were statistical differences compared with those before operation [(-2.89±4.62) mm and 1.59±0.22,respectiovely].The average leg-length discrepancy at the last follow-up was (0.78t±0.26) cm,which had on statistical differences compared with that [(0.83 ±0.33) cm]before operation.Conclusion Greater trochanteric transfer for the treatment of coxa brevis in children and adolescents could improve the clinical symptom,recover the normal anatomy of the proximal femoral,restore the hip biomechanics environment,but could uot improve the leg-length discrepancy.

OBJECTIVE:To observe the effects of Xiaoer Reganning oral liquid(XRN) on clinical symptoms and signs of children with acute upper respiratory tract infections.METHODS:161 children with acute upper respiratory tract infections were randomly divided into three groups.The patients were treated with XRN(XRN group,60 cases),Shuanghuanglian oral liquid(SHL group,41 cases) and Wei C Yingqiao granules(WCG group,60 cases),3～4 times every day for 3 days.The changes of clinical symptoms of coughing rhinorrhea,lassitude,anorexia and dyssomnia,and signs of pharyngeal congestion and tonsillar swelling were studied on the patients before and after treatment for three days,and the results of three groups were compared.RESULTS:The improvement of the clinical symptoms and signs in XRN group was better than that in the control groups(P

Objective: To evaluate the antipyretic action of Xiaoerreganning Oral Solution on acute upper respiratory tract infection of children. Methods: 161 children with acute upper respiratory tract infection were randomly divided into 3 groups. 60 patients at different age ranges in Xiaoerreganning Oral Solution group were treated with Xiaoerreganning Oral Solution at different dosags, 3～4 times every day. The treatment course was 3 days. Patients' body-temperatures before and after treatment were recorded. The temperature differences in various time points before and after treatment were computed, and compared with the results of Shuanghuanglian Oral Solution group and Wei C Yingqiao Tablets group.Results: All three drugs could reduce the patients' body-temperatures gradually, but the antipyretic action of Xiaoerreganning Oral Solution was the strongest. There were significant differences in temperature difference between Xiaoerreganning Oral Solution Group and Wei C Yinqiao Granules (in 2 or 6h after treatment and in 1 day or 3 days after treatment, P

BACKGROUND:Animal experiments have demonstrated that transplanted bone marrow stem cells (BMSCs)in the myocardial infarction region can directionally differentiate into myocardial cells with normal physiological function and promote neovascularization. Clinical studies have also showed that the cardiac function can be improved in myocardial infarction and cardiomyopathy patients after stem cell transplantation.OBJECTIVE: To observe the effect of autologous BMSCs transplantation on short-term cardiac function of patients with heart failure of ischemic cardiomyopathy.DESIGN: Self-control study.SETTING: Department of Cardiology, Xiangtan Central Hospital.PARTICIPANTS: Twenty-one patients with ischemic cardiomyopathy, including 13 males and 8 females, aged (64±6)years,who received treatment in the Department of Cardiology,Xiangtan Central Hospital of Hunan Province from March 2004 to January 2006 were retrieved. Inclusive criteria: with previous myocardial infarction at least once, B-mode ultrasonic cardiac examination showed that cardiac chamber was expanded, obvious cardiac inadequacy or stenocardia existed before stent implantation and hospitalized repeatedly, underwent percutaneous coronary artery intervention for restoring blood flow of infarcted vessel to TIMI3 degree over 3 months,but cardiac inadequacy existed to different degrees.Coronary arteriongraphy showed that no stenosis was found in the stent implanted in the coronary artery.Informed consents were obtained from all the patients.METHODS:After admission, all the patients received BMSCs transplantation based on routine drug treatment.Infarction-related arterial passage was established by percutaneous transluminal catheter technique and occluded by balloon.Isolated bone marrow stem cell suspension was injected into infarction-related arterial passage through the central cavity of catheter. ① Left ventricular ejection fractions (LVEF) and left ventricular end-diastolic diameter(LVDd)were measured before and 6 months after transplantation.② 24-hour dynamic electrocardiogram evaluation was conducted before and 6 months after transplantation under the precondition of not taking antiarrhythmic drugs. ③Clinical cardiac functional grading was conducted before and 6 months after transplantation by NYHA grading method: Grade Ⅰto Ⅳ: the higher grade, the severer symptom. ④ Adverse events and side effects were observed after operation.MAIN OUTCOME MEASURES:① LVEF and LVDd were measured before and 6 months after transplantation. ②24-hour dynamic electrocardiogram evaluation results. ③ Clinical cardiac functional grading evaluation results. ④ Post-operative adverse events and side effects.RESULTS:All the involved 21 patients participated in the result analysis.①The LVEF of patients 6 months after transplantation of BMSCs was more than that before transplantation [(54.4±6.2)％, (44.6±6.4)％,t = -5.946, P＜ 0.01], and LVDd of patients 6 months after transplantation was smaller than that before transplantation [(54.6±4.2), (60.2±4.4) mm,t = 5.306, P ＜ 0.01]. ② No new arrhythmic types appeared, and case of malignant serious cardiac arrhythmias were not increased. ③ Six months after transplantation of BMSCs, there were totally 9 patients with cardiac function of grade Ⅲ and Ⅳ, while there were 18 patients before transplantation. ④ The whole transplantation was safe.No patients were found to undergo re-examination of coronary arteriongraphy, which showed stent necrosis, due to chest pain, and no dead cases were either found.CONCLUSION:It is feasible to treat ischemic cardiomyopathy by percutaneous coronary transplantation of BMSCs,which can boost LVEF and improve cardiac function after transplantation.

Objective:To investigate the effects and underlying mechanisms of phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/NF-κB signaling pathway in human kidney-2(HK-2) cells of hyperuricemic nephropathy.Methods:HK-2 cells were cultured in vitro and randomly divided into control group and experimental group. The experimental group was induced by high uric acid (720 μmol/L) immersion for 48 h to establish a cell model of hyperuricemic nephropathy in vitro and subsequently divided into hyperuricemic group, overexpressed protease activated receptor 2 (PAR2) and knockdown PAR2 group. The expressions of PAR2, PI3K, AKT, NF-κB mRNA were measured by real-time PCR. The expressions of PAR2, PI3K, AKT and NF-κB protein were measured by Western blotting. The expressions of tumor necrosis factor-α (TNF-α), monocyte chemotactic protein-1 (MCP-1), interleukin-6 (IL-6), pro-interleukin-1β (pro-IL-1β), interleukin-1β (IL-1β) and transforming growth factor-β1 (TGF-β1) were detected by enzyme linked immunosorbent assay (ELISA). Results:(1) Compared with the control group, the expressions of PAR2, PI3K, AKT and NF-κB mRNA and protein in hyperuricemic group were significantly increased (all P<0.05), the expressions of TNF-α, MCP-1, IL-6, pro-IL-1β, IL-1β and TGF-β1 in the supernatant in hyperuricemic group were significantly increased (all P<0.01). (2) Compared with the hyperuricemic group, the expressions of PAR2, PI3K, AKT and NF-κB mRNA and protein in overexpressed PAR2 group were significantly increased (all P<0.05), the expressions of TNF-α, MCP-1, IL-6, IL-1β and TGF-β1 in the supernatant were significantly increased (all P<0.05). (3) Compared with the hyperuricemic group, the expression of PAR2, PI3K, AKT and NF-κB mRNA and protein in knockdown PAR2 group were significantly decreased (all P<0.05), the expressions of IL-6, pro-IL-1β, IL-1β and TGF-β1 in the supernatant were significantly decreased (all P<0.05). Conclusions:In the process of uric acid-induced HK-2 cell damage, uric acid significantly up-regulates the expression of PI3K/AKT/NF-κB signaling pathway by activating PAR2, leading to a marked increase in inflammatory damage. Knocking down PAR2 inhibits the expression of PI3K/AKT/NF-κB signaling pathway, which can effectively reduce the inflammatory damage of HK-2 cells.