Objective To develop a method for determination of catechins and alkaloids in brick-tea by high performance liquid chromatography(HPLC) with diode array detection(DAD),and to explore optimum extraction conditions for catechins and alkaloids components in brick-tea.Methods Catechins and alkaloids were separated and detected by HPLC.Orthogonal experiment and paired t test was carried out to compare the effect of water and alcohol as extraction solvents and to obtain optimum extraction conditions for extracting catechins and alkaloids components in brick-tea.Results Six kinds of catechins of catechin,epicatechin,gallocatechin,epigallocatechin,epicatechin gallate,epigallocatechin gallate and two kinds of alkaloids of caffeine and theobromine were separated and detected simultaneously.Calibration curves between peak areas and concentration of each component in bricktea were linear within a suitable concentration range,and coefficients of determination (R2) were between 0.9990-0.9999; spiked recoveries were from 83.78％ to 106.35％,and relative standard deviations(RSD) were between 0.50％-1.51％.The optimum extraction condition for catechins and alkaloids was 80％ ethanol,solid-liquid ratio of 1:10,temperature 80 ℃ and extraction time 30 min.Conclusion Alcohol as extraction solvents with optimal combination on HPLC,six kinds of catechins and two kinds of alkaloids are separated accurately,qualitatively,rapidly and sensitively.

The Coat protein and Maturase gene of E.coli bacteriophage MS2 was amplified by PCR,then the gene was cloned into pET32a to construct the intermediate vector pET32aCP.The conservative sequence of FMDV internal ribosome entry site(IRES) was cloned into the downstream of pET32aCP bacteriophage gene to construct the prokaryotic expression vector pCPES.The recombinant plasmid pCPES transformed into E.coli strain BL21(DE3) was induced to express with 1mmol/L IPTG.The expression products were purified by sucrose density gradient centrifugation.The expression products observed by TEM were circular viruslike particles,and the diameter of these particles was about 26nm.The stability of viruslike particles was detected,and the viruslike particles was identified by RTPCR.The results showed that the viruslike particles contain the FMDV IRES RNA and have good stability.The viruslike particles have great prospect as the standard and quality control in the area of RNA virus detection.

BACKGROUND: Schizophrenia is substantially heritable, but specific susceptibility genes remain difficult to be identified. Therefore, it is necessary to explore hereditary markers first.OBJECTIVE: To investigate the relationship between schizophrenia and related vWA allele genes based on the analysis of microsatellite DNA vWA polymorphism.DESIGN: A case-controlled study with schizophrenic patients and randomly selected population as subjects.SETTING: Ward of Dalian Seventh People's Hospital and Molecular Biological Laboratory of Dalian Medical University.between March and July 2002 at Dalian Seventh People's Hospital which specializes in schizophrenia. Schizophrenia was diagnosed according to the diagnostic standard of the third edition of "the American Diagnostic Statistical Manual for Schizophrenic Diseases", and their clinical manifestations were predominantly negative signs. Altogether 123 normal blood samples were collected from random population at the Blood Center of Dalian Red Cross. They all denied psychological ailments and severe systematic diseases, and they had no kinship with each other.METHODS: Heparin anti-coagulation blood samples were collected and PCR compound amplification was carried out with the aid of PE Profiler plus system. Then the products were subjected to electrophoresis and gene detection with ABI310 type gene analysis system so as to calculate the frequency of allele genes; Hardy-Weinberg equation law was used to make coincidence test and linkage analysis of the theoretical frequency and actual one. Schizophrenic patients and random population were compared and relative risk was calculated with RR=Pd × (1-Pc)/Pc × (1-Pd) in order to assess the statistical significance (RR: relative risk; Pd: gene frequency of schizophrenia; Pc: gene frequency of random population). RR ＞ 1 was considered of higher susceptibility while RR ＜ 1 was considered of anti-susceptibility. In this way, we could find out vWA allele genes that had susceptible linkage or anti-linkage with schizophrenic related genes.MAIN OUTCOME MEASURES: Major outcome: Correlation analysis of vWA allele genes in schizophrenic patients and random population. Secondary outcome: The coincidence of vWA allele gene frequency in patients with schizophrenia and random population with what was calculated by Hardy-Weinberg law.RESULTS: Data of the two groups were processed according to the objective and statistically analyzed.① vWA allele gene frequency in patients with schizophrenia and in random population was found to coincide with HardyWeinberg law(P ＞ 0.05).② The positive rate of vWA-14 in schizophrenic patients (17.2%) was obviously different from that in random population (33.3%) (RR=0.415, P=0.014). The positive rate of vWA-17 in schizophrenic patients (31.3%) was found to be significantly higher than that in random population (19.5%) (RR=1.866, P=0.043) while it did not differ significantly in other allele genes (P ＞ 0.05).CONCLUSION: The positive rate of vWA-14 was significantly lower in schizophrenic patients than in random population, indicating that vWA-14locus may be negatively selected in schizophrenia due to some reasons,which may be approximate to anti-schizophrenia genes. Moreover, the higher expression of vWA-17 in schizophrenic patients than in random population suggests that vWA-17 locus is correlated with schizophrenia,which may be approximate to schizophrenia-susceptibile genes.

Animals ; Brain Injuries ; blood ; cerebrospinal fluid ; chemically induced ; Endotoxins ; toxicity ; Female ; Male ; Natriuretic Peptide, C-Type ; analysis ; Neurotensin ; analysis ; Rabbits

Aged ; Humans ; Male ; Middle Aged ; Penis ; innervation ; Prostatectomy ; methods ; Prostatic Neoplasms ; surgery

Objective To study the changes of serum level of high mobility group box-1(HMGB- 1)in patients with multiple trauma in order to forecast organ dysfunction(OD)and deaths.Methods The optical densities of HMGB-1 in serum of 35 patients with multiple trauma were determined on 1st,3rd, and 7th days after trauma,and the incidence of organ dysfunction and deaths were evaluated,then analyzed statistically to learn the relation between the serum levels of HMGB-1 and deaths with an attempt of predic- ting the incident of organ dysfunction and deaths.Results (1)As OD was concerned,there was a statis- tically significant difference in optical density of HMGB-1 on 1st and 3rd days between the two groups of multiple injury patients(t=4.411,P

12 strains of H2-producing bacteria were isolated and purified from anaerobic sludge, aerobic sludge and river bottom sludge by Hungate method. A new species of high efficient hydrogen production bacterium Enterococcus sp. LG1 (registration number: EU258743 ) was studied deeply. It was showed that the Enterococcus sp. LG1 was an anaerobic and Gram-negative bacterium. Sequence analysis of this type of clones showed that it was affiliated with the genus Enterococcus and it was not reported yet in other paper at present. Meanwhile, batch tests of anaerobic fermentative hydrogen production by Enterococcus sp. LG1 were investigated by using sterilization pretreated sludge as substrate. The changes of soluble COD, protein, carbohydrate and pH value during hydrogen fermentation were monitored. It was found that only hydrogen and carbon dioxide were produced by this strain and no methane was detected during fermentation. The maximal hydrogen yield was 36.48 mL/g TCOD and the hydrogen concentration in the gas phase was 73.5%. The Enterococcus sp. LG1 was a butyrate fermentation bacteria analyzed by metabolites.

The dynamic character of acupoint has been observed extensively, but it is rarely analyzed systematically. The aim of this paper is to analyze the essence of this unique dynamic character of acupoint based on old and modern research literatures. Modern studies are summarized on qi and blood changes on the same acupoint at different times, in different physiological and pathological conditions and different syndrome types. On the basis of the literatures, the scientific base of dynamic character of acupoint is explored preliminarily. The authors propose that the dynamic character of acupoint should be focus on the modern theory of acupuncture and clinical treatment and modern preventive medicine, and the authors also put forward ideas and prospect on the essence, basis and application direction of dynamic character of acupoint.
Acupuncture Points ; Blood Circulation ; Humans ; Qi

OBJECTIVETo determine the effects of recombinant soluble complement receptor type I (sCR1) on the immune inflammatory reaction in acute spinal cord injury tissue of rats and its protective effects.

METHODSSD rat models of acute spinal cord injury were prepared by modified Allen's method. The motor function of the rat lower extremities in sCR1 group and normal saline (NS) group was evaluated by the tiltboard experiment at 12 h, 1 d, 3 d, 7 d, and 14 d. The neutrophil infiltration and C3c positive expression were observed. The myeloperoxidase activity was assessed in the injury tissue at 12 h, 1 d, 3 d, 7 d, and 14 d after injury in the two groups.

RESULTSThe motor function of rat in sCR1 group at 3 d, 7 d, and 14 d was obviously better than that in NS group (P<0.01, P<0.01, P<0.01). C3c positive expression in sCR1 group at each time point after injury was obviously less than that in NS group (P<0.01). The myeloperoxidase activity in sCR1 group at each time point after injury was obviously less than that in NS group (P<0.01).

CONCLUSIONSRecombinant soluble complement receptor type I (sCR1) can lessen the immune inflammatory reaction in acute spinal cord injury tissue and relieve secondary spinal cord injury by inhibiting the activation of the complement system.

Animals ; Disease Models, Animal ; Immunohistochemistry ; Inflammation ; Peroxidase ; biosynthesis ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Complement ; therapeutic use ; Recombinant Proteins ; therapeutic use ; Spinal Cord Injuries ; drug therapy ; enzymology ; pathology

ObjectiveTo investigate the predictive value of heart rate turbulence(HRT) in patients with diabetes mellitus (DM) after acute myocardial infarction (AMI).MethodsNinety-two AMI patients combined with DM (DM group) and 120 AMI patients without DM (non-DM group) were selected.Turbulence onset (TO) and turbulence slope (TS) were two indexes of HRT.HRT was considered positive when TO was ≥0 and TS was ≤2.5 ms/R-R.The differences in clinical data between HRT-positive and HRT-negative patients were compared.And the related risk factors after AMI were analyzed.ResultsAge,left ventricular ejection fraction (LVEF) level,renal insufficiency,LVEF＜40％,standard deviation of sinus cardiac cycle (R-R interval)(SDNN),heart rate variability (HRV) positiveand HRT indexes (TO,TS) between HRT-positive and HRT-negative patients in DM group had significant differences (P ＜ 0.05 ).Age,LVEF level,SDNN and HRT indexes(TO,TS) between HRT-positive and HRT-negative patients in non-DM group had significant differences(P ＜ 0.05).Multivariate Cox regression analysis showed that renal insufficiency (OR=4.8,95％ CI:1.8 - 10.7,P=0.008) and HRT positive (OR=3.7,95％ CI:1.5 - 8.6,P=0.070) in DM group had statistical significance.And HRT positive in non-DM group had statisticalsignificance(OR=23.0,95％ CI:5.2 ～ 86.0,P ＜ 0.01 ).ConclusionsHRT,an index of dynamic electrocardiogram,can predict the risk in patients with DM or without DM after AMI.