ObjectiveTo determine the diurnal variation,moon phase,and seasonal variation in the onset of stroke. MethodsThe time parameters,including the time,the moon phase and the season of the stroke occurrence for 905 in-patients from 1989-2001 were investigated.ResultsThere was a significant difference (D=0.155,P<0.01) for circadian rhythm in the onset of 321 cases with cerebral infarction,and the incidence frequency at dawn was much higher than that at other time,but the difference of the moon phase and seasonal rhythm in the onset of cerebral infarction were not significant(P>0.05).There were not diurnal variation,moon phase and seasonal rhythms in the onset for cerebral haemorrhage(P>0.05).ConclusionsThe cerebral infarction occur more frequently at dawn time.

Objective To investigate the impact of imaging quality of pitch on coronary CT angiography (CTA) with 64-detector row CT. Methods 566 patients were divided into four groups according to heart rate (≤ 50,51-70,71 ～ 80 and ≥ 80 bpm). Three dimensional reconstructions were used such as volume rendering (VR) ,maximum intensity projection(MIP) and curved planar reformation(CPR). Each group was divided into control group and cxperimential group randomly, using normal pitch and revised pitch respectively, and the imaging quality and influencing factons were analyzed among the four groups. Results There was significant difference in imaging quality among the four groups (P < 0.05). Each group had difference in imaging quality with normal pitch and revised pitch. Conclusions The revised pitch helps to improve the imaging quality and meet the demand of diagnosis.

Objective To observe the clinical efficacy of acupoint injection of mouse nerve growth factor in treating lumbar intervertebral disc herniation (LIDH).Method Totally 120 LIDH patients were randomized into a treatment group, control group 1, and control group 2, 40 cases in each group. Control group 2 was intervened by dehydration therapy; the treatment group was by acupoint injection of mouse nerve growth factor in addition to the intervention given to control group 2; control group 1 was by muscular injection of Trivitamin B in addition to the intervention given to control group 2. The common peroneal nerve and posterior tibial nerve conduction velocities and Visual Analogue Scale (VAS) were observed before and after treatment, and the clinical efficacies were compared among the three groups.Result The recovery and markedly effective rate was 97.5% in the treatment group, versus 92.5% in control group 1 and 85.0% in control group 2, and the rate in the treatment group was significantly different from that in control group 1 and 2 (P<0.05). Respectively 14 d and a month after intervention, the VAS scores were significantly different from that before treatment in the three groups (P<0.05). Respectively 14 d and 1 month after the intervention, the VAS score in the treatment group was significantly different from that in control group 1 and 2 (P<0.05). The common peroneal nerve and posterior tibial nerve conduction velocities a month after the intervention were significantly different from that before the intervention in the three groups (P<0.05). A month after the intervention, the posterior tibial nerve and peroneal nerve conduction velocities in the treatment group were significantly different from that in control group 1 and 2 (P<0.05). Conclusion Acupoint injection of mouse nerve growth factor is an effective way in treating LIDH.

The identification of the relationships between mutations of the bone morphogenetic protein type Ⅱ receptor (BMPR2) and pulmonary arterial hypertension (PAH) has been considered to be one of the most significant discoveries in this area in the 21st century. And BMPR2 mutation is responsible for the majority of hereditary PAH as well as some of idiopathic PAH. Furthermore, clinical and animal expreimental research over the past few years has revealed that BMPR2 signaling pathway plays a critical role in the initiation and progress of PAH, by participateing in the pathogenesis of PAH. In addition, the potential that BMPR2 signaling pathway is used as a therapeutic target is being evaluated. This review summarizes our current understanding of the role of BMPR2 mutations in PAH from the perspectives of genetics, epigenetics, inflammation as well as interactions with other significant pathways.

Objective:To study the mechanisms of neutrophil elastase (NE) induced expression of respiratory mucin MUC5AC. Methods:Using gene recombination techniques,four luciferase reporter gene plasmids containing different length of human MUC5AC gene promoter were constructed.Site-directed mutagenesis technique was used to establish mutants of Sp-l and NF-?B site in MUC5AC gene promoter; the relative luciferase activities were detected in the transfected human pulmonary A549 cells. Results:Series of luciferase reporter gene containing different sequences of human MUC5AC promotor were constructed successfully.NE could increase the expression of luciferase reporter gene plasmid containing -1300bp,-689bp and-324bp version of MUC5AC promoter in the transfected A549 cells (P

Objective To establish a HPLC method for simultaneous determination of uracil,hypoxanthine,guanosine, thymine,and( R,S)-goitrin in Isatidis Radix. Methods The determination was performed on a Hanbon Hedera C18 column (4. 6 mm×250 mm,5 μm). The mobile phase consisted of acetonitrile( A)and water( B)with linear gradient elution at the flow rate of 1.0 mL·min-1. The column temperature was 30 ℃. Detection wavelength was 254 nm. Results The linear range of uracil,hypoxanthine,guanosine,thymine,and(R,S)-goitrin was 1.97-39.40 mg·L-1(r=0.999 9),1.25-50.00 mg·L-1(r=0.999 9),0.25-10.40 mg·L-1(r=0. 999 6),2. 84-56. 70 mg·L-1(r=0. 999 9),and 0. 72-28.80 mg·L-1(r=0.999 8), respectively. The average recovery was 99. 36%(RSD=0. 91%),99. 79%(RSD=1. 12%),100. 90%(RSD=1. 71%),98. 67%(RSD=1. 50%),and 99. 33%(RSD=0. 94%),respectively. Conclusion The method is simple,accurate,reliable,reproducible and sensitive,which can be used as an effective method for quality control of Isatidis Radix.

Muscle recently has been identified as a good source of adult stem cells that can differentiate into cells of different lineages.Researchers have identified two types of stem cells in skeletal muscle.Further research is necessary to delineate the relationship between different populations of musclederived stem cells(MDSCs)and between MDSCs and other adult stem cells.The methods used to isolate these cells appear to influence the stem cell characteristics.As these efforts continue,the potential for MDSCsbased therapy for other musculoskeletal injuries,as well as for cardiac and smooth muscle injuries,is currently being explored.The behavior,biocharacteristic,isolation,differentiation and the probability of application to regenerate lost or diseased tissue of MDSCs were summarized.

Objective To construct and express a recombinant plasmid pGEX-Sj26GST of Schistosoma japonicum(Sj) in Escherichia coli(E.coli) BL21 (DE3).Methods Total RNA was extracted from Sj adult worms by RNeasy Mini kit,26 kilodalton glutathione-S-transferases of Schistosomajaponicum (Sj26GST) antigen gene was amplified by real-time PCR(RT-PCR) from the total RNA,then cloned into a prokaryotic expression plasmid pGEX1λT and transformed into E.coli BL21 (DE3) to construct pGEX-Sj26GST; BL21 (pGEX-Sj26GST) was induced with isopropyl-beta-D-thiogalactopyranoside (IPTG),and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) and Western blotting.Results The 676 bp Sj26GST gene was successfully amplified by RT-PCR and restriction enzyme double-digestion technique confirmed that Sj26GST antigen gene was successfully cloned into pGEX-1λT vector,the relative molecular mass of the expressed recombinant protein was approximately 52 × 103 by SDS-PAGE,and the amount of expressed protein was 20％ of the total bacterial proteins; the fusion protein could be recognized by sera from rabbits infected with Sj by Western blotting.Conclusions The recombinant plasmid pGEX-Sj26GST is successfully constructed and highly expressed in E.coli and the expressed fusion protein shows specific antigenicity.

AIM:To develop a method for determining cholic acid by HPLC-ELSD and GC was applied to determing muscone;in Jawei Xihuang Soft Capsule(Calculus Bovis,Moschus,Venenum Bufonis,Olibanum,Myrrha).METHODS:A C18 column(Kromasil C18,5 ?m,4.6 mm?250 mm)was used as stationary phase,the mobile phase was methanol-0.01% glacial acetic acid(73:27) at a flow rate of 1.0 mL/min.The parameters of ELSD were set as follows:evaporation temperature was 40 ℃,carrier gas(N2) pressure was 200 kPa.The GC system consisted of DB-1 capillary column(30 m?0.32 mm?0.25 ?m) and FID as the detector.The programmed temperature-GC and internal standard method were employed to determine the content of muscone.RESULTS:The linear ranges of cholic acid and muscone were in the range of 45.2 ng-904 ng and 0.05 mg/mL—0.5 mg/mL respectively.The average recoveries were 99.06% and 99.40% with RSD of 1.56% and 0.95% respectively.CONCLUSION:The method is convenient and accurate,and it can be used for the quality evaluation of Jawei Xihuang Soft Capsule.

BACKGROUND: Previous researches indicate that ADp14ARF transfecting positive tumor cell line of p53 can inhibit the proliferation; in addition, the inhibitory effect is superior to transfection negative tumor cell line of p53. Whether simultaneous transfection of p14ARF and p53 genes can increase expression and accumulation of p53 and accelerate apoptosis of tumor cells needs further studies.OBJECTIVE: To construct double plasmid expression vector plRES-p14ARF-p53 by using gene engineering so as to observe the inhibitory effect on proliferation of osteogenic sarcoma cells.DESIGN: Randomized controlled observation.SETTING: Department of Orthopaedics, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiment was carried out in the Public Laboratory Platform, Immune Researching Room, Basic Medical College, Tongji Medical College, Huazhong University of Science and Technology from January 2005 to October 2006. Human osteogenic sarcoma MG-63 cells were provided by Cell Laboratory, Immune Researching Room, Tongji Medical College, Huazhong University of Science and Technology. plRES-p53 plasmid and plRES vector containing p53total-length gene order were provided by Wuhan Jingsai Biology Company.METHODS: Based on gene engineering, p14DNA (0.5 kb) was amplified from cultured L02 cells of normal human hepatic cells into plRES vector. Recombinant plasmid plRES-p14ARF-p53 was determined with polymerase chain reaction (PCR) and restriction enzyme and transfected into human osteogenic sarcoma MG-63 cells through mediation of liposome to screen positive clones. Otherwise, cells were divided into three groups, including blank control group (MG-63cells), blank vector control group (stably transfecting plRES-neo cells) and p14ARF-p53 group (stably transfecting plRES-p14ARF-p53 cells). ① DNA content and cycle of tumor cells were measured by using flow cytometry before and after transfection. ② Reverse transcription polymerase chain reaction (RT-PCR) and Western blot were used to detect quantitative and semi-quantitative expression of p53 and p14ARF protein in tumor cells after stable transfection. ③Thiazole blue chromatometry and growth curve were used to observe proliferation.MAIN OUTCOME MEASURES: ① DNA content and cycle of osteogenic sarcoma cells; ② expressions of p53 and p14ARF protein in tumor cells; ③ proliferation.RESULTS: Double plasmid expression vector plRES-p14ARF-p53 was constructed successfully. ① DNA content and cycle of osteogenic sarcoma cells: Flow cytometry demonstrated that tumor cells mainly stayed in G1 phase after transfection. ② Protein expression: RT-PCR and Western blot indicated that p14ARF and p53 gene independently expressed in target cell mRNA and protein, respectively. ③ Cell growth: At 24, 48, 72 and 96 hours after MG-63 transfection, inhibitory rates of tumor cells were 33.43%, 69.37%, 66.19% and 75.26%, respectively, which was significant difference as compared with blank vector control group (P ＜ 0.01).CONCLUSION: Wild p53 and p14ARF can synergistically inhibit the proliferation and accelerate the apoptosis of osteogenic sarcoma cells.