AIM:To develope a reversed-phase high performance liquid chromatographic method for determing chlorogenic acid and caffeic acid in Fanhuncao Granule(Senecio cannabifolius Less). METHODS: The operation was carried out on Eclipse XDB-C_(18) column with the mobile phase consisting of a mixture of acetonitrile-0.4% phosphoric acid(12(∶)88,V/V).The flow rate of 1.0 mL/min and UV detection wavelength at 327 nm were set to determine the contents of chlorogenic acid and caffieic acid. RESULTS: There was good linear relationship between the concentrations and the peak-areas of chlorogenic acid and caffeic acid.The two kinds of standard solutions were both stable in 16 h(RSD=1.55% for chlorogenic acid,1.11% for caffeic acid) The average recovery was 100.1% and 99.8% for chlorogenic acid and caffeic acid,repectively. CONCLUSION: The method is simple,sensitive,rapid and accurate,and can be used for the quality control of Fanhuncao Granule.

Objective To investigate the correlation between hepatitis B virus(HBV)markers and AFU,ALT,GGT.Methods 570 eases of HBV-infected people were divided into 4 groups according to the different performance modes.150 healthy people were considered as control group.rate method was used to detect AFU,ALT and GGT.Estimate the differences between the groups were compared.Results The comparison between the AFU,ALT,GGT in different performance mode of HBV and the control group showed:the differences of group 1,group 2 and group 3 were statistically significant(P＜0.01),the difference of group 4 was not significant(P＞0.01).The correlation between AFU,ALT,GGT of different performance mode showed:the changes of AFU and ALT,GGT in group 1,group 2 and group 3 were positively change-related.But the changes of AFU,ALT and GGT were not clearly related.The distribution of the rise of serum enzyme activity in different performance mode showed:the AFU,ALT and GGT rised in different proportion in group 1,group 2 and group 3.Conclusions The AFU,ALT and GGT activity of HBVinfected persons should be detected at the same time,and evaluate the damage of liver cells.This will do help to predict the progression of hepatitis.

Objective To observe the effects of permanent magnetic fields with different magnetic inductions on oxidative damage to human brain microvascular endothelial cells (HBMECs) under normal and hypoxic conditions. Methods HBMECs were cultured in vitro under normal and hypoxic conditions,then divided into a control group and groups receiving magnetic induction at 8.1,16.5,20.3,26.0,27.3,62.5,110.7 and 215.6 mT.Changes in superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in each group were measured 72 h after exposure to the magnetic fields. Results Under normal conditions the SOD activities of the magnetic groups were not significantly different from that of the control group. Only the MDA content of the 215.6 mT group was statistically different (slightly higher) than that of the control group (P ＜ 0.05 ).Under hypoxic conditions,there was again no statistically significant difference in SOD activity,but the MDA content of the 110.7 mT group was significantly lower. Conclusions Under normal conditions,a 215.6 mT permanent magnetic field has an effect on MDA in HBMECs; under hypoxic conditions a 110.7 mT permanent magnetic field has such an effect.

Objective To explore the magnetic properties of permanent magnet wafers commonly used in magnetotherapy to provide a basis for the scientific and rational use of sources of magnetism.Methods A magnetometer was used to measure the magnetic induction intensities of the N and S polar centers of eight groups of magnet wafers (axial magnetization) with nominal magnetic induction intensities of 6 mT, 10 mT, 20 mT, 30 mT, 60 mT,100 mT, 200 mT and 350 mT.Results There were some differences between the nominal values and the measured mean magnetic induction intensities. In a few groups the differences were close to 20％. There were also differences within the same group. The magnetic induction intensities of the two polar surfaces were not always equal for the same magnet, and there were large differences among individual magnets.Conclusions The intensities of magnetic sources must be measured before use to ensure the consistency of magnetic induction intensities in therapeutic applications.

Objective To explore the possible roles of PI3K pathway and MAPK pathway in mediating the survival and differentiation-promoting effect of glial cell line-derived neurotrophic factor(GDNF) on midbrain dopaminergic neurons.(Methods Midbrain) slices of early postnatal rats were cultured,the slices were divided into four groups according to the different substances added into the culture medium: i.e.blank control group,GDNF group,Wortmanin plus GDNF group,and PD98059 plus GDNF group.On the 6th day,some slices were fixed,embedded and sectioned.The sections were processed for tyrosine hydroxylase(TH) immunohistochemistry,and then examined under microscope to determine the morphologic index for statistic analysis(n=6).Meanwhile,some slices were taken for Western blot to examine the expression of TH protein in midbrain slices(n=4). Results GDNF group slices showed the morphologic maturity,the density and the diameter of TH positive neurons,as well as the level of TH expression,were all significantly higher than that of the control groups.The survival effect of GDNF on DA neurons was almost abolished when the PI3K pathway was blocked with Wortmannin;while the MAP kinase and ERK kinase(MEK) inhibitor PD 98059 was added to block the MAPK pathway,the diameter of TH positive neurons was decreased significantly.Conclusion The PI3K pathway might mediate the survival effect of GDNF,while the MAPK pathway seems to be involved in the differentiation process.

Objective To observe the proliferation and differentiation of the neural progenitors in the midbrain substantia nigra(SN) of adult rat when dopaminergic neurons(DA neurons) were destroyed. Methods The Parkinson disease model(PD model) was induced in adult SD rats by steriotaxic injection of 6-OHDA into the right striatum,and then assessed by behavioral analysis to screen the qualified models.After varied survival period,the brain of model rats were perfused and fixed by 4% paraformaldehyde,moved the segment of SN,embedded with paraffin and coronally sectioned continuously.The microsections were processed by immunohistochemistry labeling separately the neural progenitors with anti-nestin monoclonal antibody,the dividing cells with anti-PCNA(proliferation cell nuclear antigen),the neuronal precursors with anti-Tuj1(?-tubulin isotype Ⅲ),and DA neurons with anti-TH(tyrosine hydroxylase).The labeled cells were counted under microscope and analyzed statistically.(Results It) was found in the right SN of PD model rats that: 1.Nestin positive(Nestin~+) cells appeared 10d after 6-OHDA injection,became abundant on 14d,declined in number on 17d,and disappeared on 21d.2.Weakly positive PCNA(PCNA~+) cells appeared on 7d.PCNA~+ cells were abundant on 14d,decreased in number from 21d,with only a few positive cells noticed on 28d.3.Tuj1 positive cells appeared in small number on 10d,became abundant on 14d,decreased in number from 17d and dropped nearly to zero on 21d.4.The number of TH positive neurons was significantly less than the normal control(by 24%) on 7d,and became even less as time elapsed.Conclusion When 6-OHDA is injected into the striatum of adult rats to cause degeneration and death of the DA neurons there,there would be a certain period of time in which a number of neural progenitors will be induced to proliferate actively and differentiate toward neuronal cells(except DA neurons).

Mortality of patients with malignant middle cerebral artery infarction (mMCAI) is as high as 41％ to 78％ with the conservative treatment in the intensive care unit.Malignant edema caused tentorial herniation is the most common cause of resulting in the death of patients.Some evidence supports the hemicraniectomy for the treatment of patients with mMCAI.The patients must receive supportive and adjuvant therapies before surgical decision making.Therefore,the prediction and intensive care of mMCAI have a very important significance.This article reviews the predictive factors and intensive care of mMCAI.

Objective:To observe the stability of thymosin ?_1 microspheres under preparation conditions.Methods: Using HPLC method,we investigated the influence of different temperatures(-20 ℃, 6-8℃,60℃),sonification time(10-60 s) and pH values (pH 4.0,7.0 and 10.0) on thymosin ?_1 stability.Results: Under the conditions of frozen environment(-20℃) and refrigeration(6-8℃), thymosin ?_1 remained stable for 30 d.Thymosin ?_1 did not degrade in 60℃ water bath for 12 h or in phosphate buffer solution(PBS,pH 4.0) under 37℃ for 14 h.The sonification time within 60 s was found to be safe for the preparation.Conclusion: Thymosin ?_1 is stable under these preparation conditions.

Objective To understand the status of mycoplasma infection and drug susceptibility to provide reference basis for clinical rational drug use.Methods The reagents kits produced by Shanghai Aopu Biopharmaceutical Co.,Ltd.were adopted to per-form the mycoplasma culture,identification and susceptibility test.Results Among 1 745 specimens,896 cases of mycoplasma in-fection were detected with the total detection rate of 51 .35%,including Ureaplasma urealyticum (Uu)infection in 841 cases (48.19% ),Uu combined Mycoplasma hominis(Mh)infection in 92 cases(5.27% )and Mh infection in 17 cases(0.97%).The de-tection rate was 9.46% in males and 41 .89% in females,the infection rate in females was obviously higher than that of males.The results of drug susceptibility test showed that the sensitive rate to doxycycline,josamycin and minocycline was higher than 95%, while the resistant rate to norfloxacin and lincomycin was higher than 75%.Conclusion Mycoplasma infection in this hospital is dominated by Uu.Doxycycline,josamycin and minocycline can be used as the empirical medication to treat mycoplasma infections in this hospital.But before clinical treatment,the mycoplasma culture and the drug susceptibility test should be performed as possible for rational and standardized medication.

Objective To establish a kind of detection technique of nucleic acid based on surface plasmon resonance(SPR) and to set up the foundation of real-time, online space microbial detection. Methods A portable online bio-molecules analyzer based on SPR biosensor was applied. The probe was mercapto-modified at the 5’ end and immobilized on the sensor surface. Then the target sequences in the solution were monitored and sensitivity, specificity and reproducibility of the method were investigated. Results The results showed that detection method with good specificity and sensitivity could realize online detection of target sequence. The system could detect 2.3 nmol/L target sequence, CV value of nine detections was 3.5% and that of thirty detections was 14.7%. Conclusion The established nucleic acid detection method has the advantage of high sensitivity, good specificity and reproducibility, which can be applied in the field of nucleic acid detection.