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 observe the biocompatibility of a biomimetic designing of a multi-grade compositions in repairing articular cartilage and subchondral bone in animal bodies and repair the fullthickness defects in articular cartilage with the compositions and to study the regenerated cartilage histomorphologically. Methods Biocompatibility study: Acute general toxicity test, Haemolysis test, subcutaneous implantation test and chronic toxicity test. Articular cartilage defects repaired experimental study :The models of defects in articular cartilage were made artificially in both condylus lateralis femoris of mature rabbits. Implanted with the biomimetic designing of a multi grade compositions randomly at one side as the experimental group and the other side were untreated as the control group. The rabbits were killed at 4, 6, 8and 12 weeks after operation, respectively, with 6 ones at each time, and the macroscopic, histological, ultrastroctural examinations and semi-quantity cartilage scoring employing Wakitanifa repaired cartilage value system were performed. Results Biocompatibility study: (1) The rabbits' weight in experimental group kept growing .(2) Haemolysis rate of rats to different concentrations of diffusion solution was＜5%.(3) In chronic toxic reaction, rabbits' liver and kidney function was not different compared with the control groups at 12weeks and the index before operation. Articular cartilage defects repaired experimental study: 4-8 weeks after operation, the defects in the experimental group were partly filled with hyaline cartilage. Twelve weeks after operation, the defects in the experimental group were completely filled with mature hyaline cartilage.However, fibrous tissues were seen in the control group all the time. At 4, 6, 8, and 12 weeks postoperatively, the Wakitanifa cartilage scores were (7.60±0.98), (5.69±0.58), (4.46±0.85) and (4.35±0.12), respectively,in the experimental group and (10.25±1.05), (9.04±0.96), (8.96±0.88) and (8.88±0.68), respectively, in the control group. Differences between the control group and the experimental group were significant. Conclu sion The biomimetic designing of a multi-grade compositions has good biocompatibility and may induce cartilage regeneration to repair the full-hickness defects of articular cartilage.

BACKGROUND: CpG oligonucleotide has been shown to strengthen the function of peripheral blood mononuclear cells (PBMCs), but its effects on type 1 diabetes mellitus has been rarely reported. OBJECTIVE: To investigate the effects of CpG oligonucleotide on the expression of interferon γ (IFN-γ), interleukin (IL) -12 and IL-10 in PBMCs in patients with type 1 diabetes mellitus versus healthy controls. METHODS: PBMCs were isolated from patients with type 1 diabetes mellitus and healthy controls and then cultured in RPMI-1640 with non-stimulator (control group) and CpG oligonucleotide (CpG oligonucleotide group), respectively. The mRNA expression of IFN-γ, IL-10, and IL-12 in PBMCs was detected by reverse transcription-polymerase chain reaction. RESULTS AND CONCLUSION: mRNA expression of IFN-γ and IL-10 was significantly lower in patients with type 1 diametes mellitus than in healthy controls (P < 0.01). In the CpG oligonucleotide group, the mRNA expression of IFN-γand IL-12 was significantly higher than in the healthy control group (P < 0.01), but the mRNA expression of IL-10 was similar to that in the healthy control group (P > 0.05). These findings demonstrated that CpG oligonucleotide can promote the production of IFN-γ and IL-12 in PBMCs of type 1 diametes mellitus.