Objective: To establish an ultra performance liquid chromatography(UPLC)method for determination of prot- amine peptide analogul(R15),and investigate the metabolism in biological samples of R15 in vitro by the method. Methods: Chro- matographic separation was performed by pumping the mobile phase into a BioBasic C 18 (2.1 mm×100 mm,5 μm). The mobile phase was composed of 0.05% trifluoroacetic acid-water and 0.05% trifluoroacetic acid acetonitrile(98:2,V/V),using 10% trifluoroacetic ac- id-acetonitrile-water as precipitant. The stability of R15 in rat whole blood,liver and kidney homogenate was studied. Results: The in- tra-and inter-batch relative standard deviations (RSD)were less than 15%,the relative errors (RE)were in the range of ±15%. The range of calibration curve was 25-1000 μg/ml with a good correlation coefficient. The extraction recoveries of R15 were 99.43-100.43%. R15 was stable under various experimental storage conditions. Half-lives of R15 in rat liver,rat kidneys,rat blood,beagle dog blood and human blood were 27.4 min,9.1 min,15.6 min,11.9 min and 21.0 min,respectively. Compared with the control group without inhibi- tor,the inhibitory rates of EDTA,pepstatin and benzamidine HCl protease inhibitor in the experimental group was 92.7%,41.3% and 33.0%,respectively. Protamine analogue peptide was mainly metabolized by metal carboxyl peptidases. Compared with the control group,AEBSF,leupeptin and trypsin protease inhibitors had significant difference,but the inhibitory rate was low. Conclusion: The method is specific,sensitive,accurate and reliable. The metabolism of R15 in rat kidney is the fastest. The degradation of R15 in whole blood is possibly related to metal carboxypeptidase.

BACKGROUND: It is manifested in epidemiology and clinical observation that lipoprotein (a) is a new risk factor of cerebrovascular disease and is closely related to cerebral ischemic stroke.OBJECTIVE: To discuss the relationship between levels of high serum lipoprotein (a) and stroke.DESIGN: Case controlled analysis.SETTING: Neurological Institute of Xiangya Hospital of Central South University.METHODS: Totally 294 patients with stroke were selected from Neurological Department of Xiangya Hospital of Central So, uth University between September 1999 and March 2002. Of them, 159 cases were regarded as cerebral infarction group and other 135 cases as acute hypertensive cerebral hemorrhage group. In cerebral infarction group, 109 patients had atherosclerotic cerebral infarction and 50 patients had lacunar cerebral infarction, and 94patients with contimuous health examination were regarded as health examination group. Serum lipoprotein (a) in each group was assayed with "sandwich enzyme-linked immuno-sorbent assay". According to whether the value of lipid was normal or not, patients with atherosclerotic cerebral infarction and acute hypertensive cerebral hemorrhage were divided into two groups. Comparisons between the two groups were assayed with single sample t test, and multiple liner regression was used to assay whether sex, hypertension and value of lipid were related to the level of serum lipoprotein (a).MAIN OUTCOME MEASURES: ① Comparisons of serum lipoprotein (a)among atherosclerotic cerebral infarction group, lacunar cerebral infarction group, acute hypertensive cerebral hemorrhage group and health examination group. ② Correlated analysis between serum lipoprotein (a) and lipid.RESULTS: Among 294 patients, 94 cases in control group entered the final analysis. ① Comparisons of serum lipoprotein (a) among atherosclerotic cerebral infarction group, lacunar cerebral infarction group, acute hypertensive cerebral hemorrhage group and health examination group: Levels of serum lipoprotein (a) in atherosclerotic cerebral infarction group and cerebral hemorrhage group were higher than those in health control group (P ＜ 0.05), and concentration of lipoprotein (a) in atheroosclerotic cerebral infarction group was increased as compared with that in acute hypertensive cerebral hemorrhage group (P ＜ 0.05). Also, level of lipoprotein (a) in lacunar cerebral infarction group was a little higher than that in control group,but the difference was not significant (P ＞ 0.05). ② Correlated analysis between serum lipoprotein (a) and lipid: Levels of lipoprotein (a) in both normal lipid group and abnormal lipid group were assayed with single sample t test, and the results showed that levels in the two groups were similar (P ＞ 0.05). Multiple liner regression was used to assay whether sex,hypertension and value of lipid were related to level of serum lipoprotein (a).CONCLUSION: Levels of lipoprotein (a) may be an independent risk factor for cerebral hemorrhage and atherosclerotic cerebral infarction.

Objective: To investigate the effect ofintestinal trefoil factor (ITF) on the transcriptional activity of ITF promoter and to explore the regulatory mechanismofJanus kinase/signal transducersand activators of transcription(JAK/STAT) on ITF promoter. Methods: The 5' flanking sequence of the ITF gene was cloned from human whole blood genomic DNA by PCR. ITF promoter fragment was cloned and inserted into the pGL3-Basic vector to construct recombinant vector. ITF promoter vector was stimulated with ITF at various concentrations and the luciferase activity was measured. The JAK-STAT3 signal transductionpathway was then blocked by a speciifc inhibitor AG490 to determine the signal pathway involved in ITF promoter activity. Results: Restriction endonuclease analysis and DNA sequencing confirmed that the recombinant plasmid, containing ITF promoter, was constructed successfully. After transient transfection, the activity of ITF promoter was increased signiifcantly in the presence of ITF (P<0.05). Blockage of the JAK-STAT3 signal transduction pathway with AG490 signiifcantly reduced the ITF promoter activity (P<0.05). Conclusion: ITF increases the transcriptional activity of ITF promoter via the JAK-STAT3 signal transduction pathway.

BACKGROUND: There has been increasing attention in the study of adolescent idiopathic scoliosis (ALS)and bone metabolism, which is accompanied by osteopenia and osteoporosis. Interleukin(IL) -6, a cytokine that strongly promotes bone resorption, participates in the regulation of bone metablism.OBJECTIVE: To explore the relationship between changes of bone mineral density(BMD) and serum IL-6 content in AIS.DESIGN: A controlled non-randomized concurrent study involving patients with AIS and and healthy volunteers.SETTING: The clinic and wards of the department of spinal surgery of a university-affiliated hospital.PARTICIPANTS: Thirty-six patients with AIS(6 males and 30 females aged 12 to 18 years) were treated in the Department of Spinal Surgery, Drum Tower Hospital, Medical College of Nanjing University from July to October 2003, who had a Cobb angle ranging from 34° - 109° and curvature of the thoracic spine. Thirty-six healthy adolescent volunteers(7 males and 29 females within the range of 13 - 18 years old) served as the control group.METHODS: The BMD was measured at L2- L4 and the proximal femur;(including the femoral neck, greater trochanter and Ward' s triangle) by dual-energy X-ray absorptiometry and serum IL-6 determined by enzyme-linked immunosorbent assay(ELISA).MAIN OUTCOME MEASURES: ① BMD of the lumbar spine and femur ② Comparison of serum IL-6 content of all the subjects.RESULTS: The BMD of AIS patients at L2- L4, femoral neck, greater trochanter and Ward' s triangle was 0.79±0. 12, 0.78±0. 12, 0.65± 0. 10, and 0. 69 ± 0. 13 g/cm2, respectively, all significantly lower than the corresponding measurements of the control group(1.09 ± 0. 11, 0.95 ± 0. 11,0. 78 ± 0. 10, and 0. 88 ± 0. 11 g/cm2, respectively, P ＜ 0. 001), whereas the serum IL-6 content in the patients was significantly higher than that in the control subjects ( P ＜ 0. 005). The changes of BMD at the lumbar spine and the three sites of femur were negatively correlated with serum IL-6 in AIS patients( P ＜ 0. 001 ), but not so in the control group( P ＞ 0.05).CONCLUSION: The BMD is decreased and serum IL-6 elevated in patients with AIS, and excessive secretion of IL-6 might be one of the important factors of osteopenia in AIS.

To study the influence of foreign plasmid DNA on spleen metabolism in immune-stimulated mice via the gastrointestinal tract. Mice were oral administered by pipette with 200?g of plasmid pcDNA3 and spleen were isolated at 4h and 18h after oral administration. Total RNA were extracted from spleen and spleen gene expression profile of Balb/c mice was analyzed by using Affymetrix oligonucleotide genechip after oral plasmid pcDNA3 administration. Functional cluster analysis was conducted by Genmapp and MAPPFinder software. By functional cluster analyzing the genes which were up-regulated more than twofolds, Genmapp results showed that plenty of metabolic pathway were induced in spleen after oral administration of plasmid pcDNA3. These metabolic process included purine metabolism, pyrimidine metabolism, protein synthesis, cholesterol synthesis, fatty acid synthesis, Glycolysis, TCA cycle and mitochondria oxidative phosphorylation pathway. The similar results also took place at 18h after oral administration. The result indicated that foreign plasmid DNA can modulate metabolism process in spleen of mice via the gastrointestinal tract, and may help understand the mechanism of action of foreign plasmid DNA uptaked via the gastrointestinal tract.

We used 4 methods,such as ultrasonic crush(UC),ultrasonic rinse(UR),whorl surge(WS)and rubbing(RU),to isolate epiphytic bacteria from red alga Gracilaria lemaneiformis.Then,we counted bacteria numbers,detected bacterial species,observed bacterial configuration and characteristic of cell wall.Compared with these methods and with different treatments in one method,the results were drawn:the UR and RU were inferior in all methods to isolate bacterial numbers and species,the UC and WS were better,especially,the treatment 30W 30s of UC was the best in experiment,which isolated 12 of 16 bacterial species,and got 1.75 10~6 cells per gram wet weight G.lemaneiformis.

Objective To explore the relationship between diet intake and diabetes mellitus(DM) and impaired fasting glucose (IFG) in adult residents in Guangxi,so as to provide scientific basis for dietary prevention.Methods 2 281 people(1 020 from urban areas and 1 261 from rural areas) aged 18 years and above were selected from 4 cities and 4 countrysides in Guangxi through a multistage stratified random sampling.The investigation included the meal investigation,medical examination and blood assay. Results Total 37 people(26 from urban and 11 from rural) suffered from DM and 26 people(15 from urban and 11 from rural) had IFG,the general prevalence rate of Impaired Glucose Metabolism(IGM) was 2.8%(4.0% for urban and 1.7% for rural);It showed that the prevalence of IGM in city was obviously higher than that in the countryside(P

Background The aspheric intraocular lenses(IOLs)can reduce ocular spherical aberration to some degree.However,the clinical effect depends more on the IOL proper alignment.It becomes more important to study the IOL position in eye,Objective This study was to analyze the position alteration of IOL after phacoemulsification combined with implantation of one-piece soft IOLs.Methods In this prospective control study,80 eyes of 40 patients with age-related cataract were enrolled.The phacoemulsification with IOL implantation was performed in all the eyes.Decentration and tilt of IOL in the nasal superior,superior temporal,inferior temporal and nasal inferior quadrants(the intersection point of the system optical axis and the IOL maximum cross plane were regarded as the ordinate origin)were measured by rotating Scheimpflug camera(Pentacam Oculus)in 3 months postoperatively under the mydriasis condition.Written informed consent was obtained from each subject prior to this trial.Results In the right eye group,the IOL decentered toward temporal in 26 eyes(65％)and infratemporal in 16 eyes(40％).IOLs tilted temporally in the horizontal plane in 37 eyes(92.5％)and tilted inferiorly in the vertical plane in 34 eyes(85.0％).In the left eye group,IOLs decentered temporally 33 eyes(82.5％)and 20 IOLs (50％)infratemporally,IOLs tilted temporally in the horizontal plane in 37 eye(92.5％)and 36 IOLs(90％)tilted inferiorly in the vertical plane.There was no statistical difference for the intercomparsion of horizontal/vertical decentration in various quadrant in the right eye(F =0.221,0.792,P＞0.05).The obvious elevated horizontal decentration was found in the supertemporal and infratemporal quadrants compared with supernasal quadrant in the left eyes but there was no significant difference in the vertical decentration among 3 quadrants(F=0.576,P＞0.05).Decentrations were positively correlated with the tilt in both horizontal and vertical plane(right eye horizontal plane:r=0.374,P=0.002;right eye vertical plane:r=0.402,P=0.001 ;left eye horizontal plane:r=0.377,P=0.002；left eye vertical plane:r=0.347,P=0.002).Conclusions The one-piece soft IOLs(Adapt AO)decenter toward temporal mostly in 3 months after surgery,especially infratemporally in the eye.And the optical axis of the IOL tilt toward infratemperol mostly in both right and left eyes.The decentration and tilt are consisted in the corresponding direction between the right and left eyes.The position of the IOLs showed mirror symmetry between right and left eyes.The IOLs decentration show the positively correlation to tilt whatever in horizontal and vertical plane.

BACKGROUND:Whether controling of post-injury inflammatory response combined with neural stem cel (NSC) transplantation can improve the curative efficacy for spinal cord injury stil remains unclear. OBJECTIVE:To investigate the repair of spinal cord tissue, myelin regeneration, axon regeneration, motor function recovery and the possible mechanism after early application of tumor necrosis factor α antagonist (Etanercept) combined with tyrosine kinase C (TrkC) gene-modified NSC transplantation. METHODS:TrkC-overexpressed NSCs (TrkC-NSCs) were constructed by lentiviral transfection technique. The rat models of spinal cord transection injury were prepared, and then subjected to Etanercept combined with TrkC-NSCs transplantation. The number of neurons and neuroregeneration after injury were measured by Nissl's staining, immunofluorescence and western blot. The rat motor function was detected by Basso Beattie Bresnahan score and evoked potential. The myelin regeneration was detected by electron microscopy and toluidine blue staining. RESULTS AND CONCLUSION:Compared with the other groups, the Etanercept combined with TrkC-NSCs transplantation group had more survived anterior horn motor neurons at 28 days after injury, more myelin-encapsulated axons, higher Basso Beattie Bresnahan score, greater amplitude of the evoked potentials, and relatively shorter latency (alP < 0.05). These findings indicate that early application of tumor necrosis factor-α antagonist combined with TrkC-NSCs transplantation after spinal cord injury in rats can effectively promote myelin regeneration, axon regeneration, and further promote motor function recovery.

Objective:To investigate the effects of the β-casomorphin-7 on in vitro and in vivo mice splenic lymphocyte and macrophage nitric oxide production.Methods:Splenocytes proliferation assay and nitrite determination were employed for the studies of effects of β-casomorphin-7 on mice splenic lymphocyte and macrophage following in vitro stimulation of β-casomorphin-7 and in vivo intrapetitoneal administration and drinking β-casomorphin-7 solution administration.Results:In vitro study, β-casomorphin-7 showed a stimulation as well as a suppression of lymphocyte proliferation and significantly(P＜0.01) suppressed the production of nitric oxide. In vivo study, β-casomorphin-7 actions on lymphocytes and macrophages were accordant in intraperitoneal administration and drinking β-casomorphin-7 solution administration. β-casomorphin-7 significantly(P＜0.01) increased in splenic lymphocyte proliferative response and suppressed nitric oxide production in peritoneal macrophages.Conclusion:The present study indicates that the β-casomorphin-7 has the immunomodulatory effects and β-casomorphin-7 may be permeable into peripheral blood intact in mice of 2-3 weeks of age.