Objective To appraise determination of serum type Ⅳ collagen protein with latex-enhanced immunoturbidimetry.Methods The evaluation test was performed including precision, linearity determination, prozone reaction, correlation, recovery test and interference test.Results The intra-batch CV% was within 2.47%-3.48%, inter-batch CV% was within 3.46%-4.07%, and recovery rate was within 96.2%-99.3%. There was a good linearity, and the tolerance limit was 800 ng/mL.Regression equation:Y=1.011 8X-1.788 3;detection correlation of different concentrations of standard preparations:r2=0.999 9,Y=1.003 1X-1.236 2.The assay had a very good anti-interference performance to bilirubin, cruorin, and triglyceride without exception.Conclusion Determination of serum type Ⅳ collagen protein attains the demand of the clinic with latex-enhanced immunoturbidimetry in fully automatic biochemical analyzer.

AIM: To observe the effects of folic acid (FA) on antioxidant enzyme, nitric oxide synthase (NOS) and nitric oxide (NO) in ovariectomized (OVX) rats.METHODS: Forty three-month-old female SD rats were randomly divided into 5 groups:sham group, OVX group, diethylstilbestrol group (0.03 mg· kg-1· d-1), low-dose FA group (5 mg· kg-1· d-1) and high-dose FA group (20 mg· kg-1· d-1).Gastric gavage started 1 week after operation and lasted for 10 weeks.The rats in sham group and OVX group were given distilled water instead of FA as controls.At the end of the 10th week, the L5 vertebra and right femur were removed for determination of bone mineral density (BMD).The bone homogenates were made using the L3 and L4 vertebrae.The levels of the total antioxidant capacity ( TAC) , glutathione peroxidase ( GSH-Px) , malondialdehyde ( MDA ) , NOS and NO were detected in plasma and bone homogenates.RE-SULTS:Compared with sham group, the BMD levels in L5 vertebra and right femur and the levels of GSH-Px and NO in the plasma were all decreased.The levels of TAC, GSH-Px, NOS and NO in the bone homogenates were also decreased, while the MDA concentration was increased in OVX group (all P<0.01).Compared with OVX group, the levels of TAC, GSH-Px, NOS, NO and BMD of the L5 vertebra and right femur were all increased, while the MDA concentration was de-creased in high-dose FA group (all P<0.01).CONCLUSION:In female SD rats, ovariectomy leads to a significant re-duction of antioxidant enzyme, NOS and NO levels.Oxidative stress is possibly involved in the development of osteoporo-sis.Protection against osteoporosis by high-dose FA may be linked to improvement of antioxidant enzyme activity, the levels of NOS and NO as well as a reduction of oxidative stress in ovariectomized rats.

Objective To investigate the protective effect of folic acid(FA) on osteoporosis in ovariectomized(OVX) rats.Methods Forty three-month-old female SD Rats were divided into 5 groups, sham operation group, OVX group, diethylstilbestrol group(0.03mg·kg-1·d-1),low dose FA Group (5 mg·kg-1·d-1),and high dose FA group (20 mg·kg-1·d-1).Gastric gavage in each group was started from one week after being ovariectomized and lasted 10 weeks. Sham operation group and OVX group were treated with solvent. The rats were sacrificed at the end of 10th week after treatment. The total homocysteine(tHcy) in plasma, alkaline phosphatase(ALP), and tartrate-resistant acid phosphatase(TRACP) activity of bone homogenates were measured. The bone mineral density(BMD) and bone biomechanics were determined using L5 vertebrae and right femur. The bone tissue slices were made with L6 vertebrae and left femur and HE stained, and then the histomorphology was observed. Results Compared with sham operation group, plasma tHcy level was significantly increased(P＜0.01), BMD of lumbar vertebrae and femur was remarkedly decreased in OVX group(all P＜0.01). Plasma tHcy concentration was negatively correlated with lumbar BMD(r=-0.359, P=0.040). Plasma tHcy level in both groups treated with folic acid was significantly reduced(all P＜0.01). The ALP concentration in bone homogenates was higher, the TRACP concentration in bone homogenates was lower, and BMD and bone biomechanics of lumbar vertebrae and femur were increased in high dose FA group than those in OVX group(all P ＜0.01). Conclusions In OVX rats hyperhomocysteinemia existed and was involved in the development of osteoporosis. Folic acid could protect OVX rats from osteoporosis, due probably to improved homocysteine metabolism.

AIM: To establish HPLC for the determination of icariin and adenosine in Cordyceps Vigorine Tablet. METHODS:MetaChem Polaris ODS C18-A column(4.6 mm × 250 mm, 5 μm) was adopted; acetonitrile-water was the mobile phase; detection wavelength was set at 272 nm for icariin and at 260 nm for adenosine; column linearity within the range of 0. 374-2. 246 μg for icariin and 0. 348-1. 392 μg for adenosine, respectively. The average recovery and RSD were found to be (99.4 ± 1.84) % for icariin ( n = 6), (99.63 ± 1.29 ) % for adenosine,respectively. CONCLUSION: The method is convenient, fast, reliable to operate and suitable for the quality control of Cordyceps Vigorine Tablet.

This study is to investigate the protective effect of astaxanthin against injured hepatocyte L-02 cells induced by sodium azide (NaN3) and reveal the possible mechanisms. Hepatocyte L-02 cells were exposed to 100 mmol.L-1 NaN3 with various concentrations of astaxanthin pre-incubated, then the cell viability was measured by MTT method; The level of reactive oxygen species (ROS) was determined by DCFH-DA method; The changes of mitochondrial membrane potential (MMP) and apoptosis ratio were detected by JC-1 method and Annexin V-FITC/PI double stain method, respectively. Results showed that after cells were exposed to 100 mmol.L-1 NaN3 for 3 hours, the cell viability significantly decreased; ROS level and the percentage of late phase apoptosis increased obviously; MMP was also declined. When cells were pretreated with astaxanthin, the cell damage and late phase apoptosis ratio reduced and MMP was maintained. However, the level of ROS showed insignificant decrease (P>0.05). The beneficial concentration of astaxanthin in improving cell viability and MMP was not in a dose dependent manner and the most effective of which was 0.10 nmol.L-1 (P<0.01). In order to reveal its possible non-antioxidant mechanism, mitochondrial membrane was imitated and H+ transferring function of astaxanthin was also detected by bilayer lipid membrane (BLM) method. Results showed that 2.0% astaxanthin could transfer H+ efficiently. These suggested the mechanisms of astaxanthin in protection of hepatocyte L-02 cells not via its ROS quenching capability but via its H+ transferring function, which improved the mitochondrial function and had the sequence biology effects.

Objective: To study the inhibition effect of agaro-oligosaccharide (AOS) on neovascularization and its mechamism. Method: The anti-angiogenic effect in vivo was evaluated on chicken chorioallantoic membrane (CAM) model. Cytotoxic activity of AOS was demonstrated by inhibition of several human cell lines by MTT assay. The apoptosis of HUVECs induced by AOS was examined by Hoechst staining assay and quantified by flowcytometry. Results: In CAM, AOS caused dose-dependent decrease on the vascular density and adversely affected capillary plexus formation. Different cytotoxic sensitivities were observed for AOS towards several kinds of cell lines, and HUVECs were more sensitive. Moreover, the growth inhibitory activity was correlated with induction of apoptosis. Flowcytometric analysis also revealed that AOS arrested the cell cycle progression at S phase. Conclusion: Agaro-oligosaccharide possesses the anti-angiogenic effects, which are associated with apoptosis induction of endothelial cells.

Objective To observe any differences in motor cortex excitability between Parkinson's disease (PD)patients and patients with multiple system atrophy (MSA) and to explore whether motor evoked potentials (MEPs) can be used as an electrophysiological indicator for differentiating the 2 diseases.Methods Thirty-four PD patients, 22 MSA patients and 15 age- and sex-matched healthy control subjeets were included in this study. Relaxed motor thresholds (RMTs), central motor conduction time (CMCTs) and MEP amplitudes (AMPs) were recorded in all three groups. The relationships of RMT, CMCT and AMP with the severity of the disease were observed.Results Average RMT in the PD group was significantly lower than that in the MSA and control groups. Average RMT in the MSA group was also significantly lower than that in the control group. There was no significant difference among the three groups with regard to CMCT. AMP in the PD group was significantly higher on average than in the MSA and control groups, but there was no significant difference between the MSA and control groups. RMT decreased and CMCT shortened progressively with the severity of the disease in the PD group, but not in the MSA group.Conclusions There were differences in motor cortex excitability between PD patients and MSA patients. MEP RMTs and CMCTs may be valuable for identifying PD and MSAc but the clinical significance of the amplitude differences remains to be further explored.

Objective To study the effects of repeated transcranial magnetic stimulation(rTMS)on Parkinson's plus syndrome(PPS).Methods Fifteen in-patients with PPS were studied between 2005 and 2008.The patients received 1 Hz rTMS at an intensity 30%over the threshold.The rTMS was applied on the hand representive area of the bilateral first motor cortex,50 stimulations on each side,5 arrays,for 5 min,once daily for 15 d.Hamilton's depression scale(HAMD),Hamilton's anxiety scale(HAMA),the unified Parkinson's disease rating scale(UPDRS,which can be subdivided into UPDRS Ⅰ,UPDRS Ⅱ and UPDRS Ⅲ),an activities of daily living scale(ADL),the mini-mental state examination(MMSE)and motor evoked potential(MEP)were assessed before and immediately after 15 d of rTMS treatment. Results Average HAMD,HAMA,UPDRS,UPDRS Ⅱ and UPDRS Ⅲ scores all decreased,and ADL scores increased significantly after treatment,while UPDRSⅠand MMSE scores were unchanged before and after treatment.No significant changes in resting motor threshold or central motor conduction time of the MEP were observed after rTMS treatment. Conclusion Clinical symptoms of PPS patients improved after rTMS treatment and side effects were few.Depression,anxiety,motor function and ability in the activities of daily living improved greatly.Repeated transcranial magnetic stimulation is a potential treatment for PPS patients.There may be no correlation between the effective mechanism of rTMS and cortex excitation.

Objective To evaluate the changes in and the regularity of brainstem evoked potentials (BA-EPs) in Parkiuson's disease (PD) as an objective criterion for early diagnosis and assessment. Methods Thirty-five healthy SD rats were divided into two groups at random. Twenty-two rats were in the experimental group and 13 in the control group. The rats were injected with 8 μg of 6-OHDA solution in the right substantia nigra pars compacta (SNc) and the right ventral tegmentum area (VTA) to create a PD model. The BAEPs of the rats in the experimental group were recorded in a quiet shielded room before the 6-OHDA injection, and one week and two weeks after injec-tion. The control group rats were injected with saline (Ns) and their BAEPs were recorded at the corresponding times. One week and two weeks later, the model rats were injected with apomorphine (APO) and their rotating cycles were counted. Results The Ⅱ , Ⅳ, andV PLs and the Ⅲ-Ⅴ IPLs on the fight ears of the experimental group were prolonged significantly compared with the control group one week after APO injection. There was no significant differ-ence in the BAEPs of the left ears after the first week. After two weeks, the Ⅱ , Ⅳ, and Ⅴ PLs and the Ⅲ-Ⅴ, and Ⅰ-Ⅴ IPLs of the right ears in the experimental group were prolonged significantly compared with the controls and the Ⅳ, and Ⅴ PLs and the Ⅲ -Ⅴ , and Ⅰ-Ⅴ IPLs on their left ears were prolonged significantly. Conclusion In the early course of a PD model in rats, their BAEPs show abnormal changes, which indicates that BAEP could be an ob-jective criterion for early diagnosis and assessment of PD. BAEP may serve as one index of damage in PD. The Ⅲ-Ⅴ PL and Ⅰ-Ⅴ iPL are sensitive indices of PD.