The content of benzyl isothiocyanate (BITC) which as the enzymatic hydrolysis product of benzyl glucosinolate through thioglucosidase was determined by HPLC. The content of benzyl isothiocyanate (BITC) which as the enzymatic hydrolysis product of benzyl glucosinolate through thioglucosidase was determined by HPLC. The chromatography condition was as follows: Kaseisorb LC ODS 2000 (4.6 mm x 150 mm, 5 min) column with the mobile phase of acetonitrile(A)-water( B) under gradient elution (0-5 min, 3%-8% A; 5-9 min, 8%-48% A; 9-23 min, 48%-62% A; 23-28 min, 62%-99% A); the flow rate was 1.0 mL x min(-1) with 10 microL injection volume; detection wavelength was 246 nm and temperature of column was 40 degrees C. The content of benzyl glucosinolate was in the range of 10.76-17.91 g x L(-1). The method is simple, accurate and good reproducibility which can be used for the determination of benzyl glucosinolate in Lepidium meyenii, effectively.
Chromatography, High Pressure Liquid ; methods ; Glucosinolates ; analysis ; Lepidium ; chemistry ; Plant Extracts ; analysis

Objective To establish rSAG1-IgM-ELISA with purified rSAG1 fusion protein for immunodiagnosis of toxoplasmosis. Methods The rSAG1 fusion protein was purified by Ni 2+ column. The ELISA plate was coated with different concentrations of rSAG1, reacted with pooled positive and negtive human sera. Goat anti-human IgM conjugated to horseradish peroxidase was used as the second antibody. The appropriate detecting condition of the rSAG1-IgM-ELISA assay was determined by orthogonal experiment. The reproducibility, sensitivity and specificity of the assay were assessed. Thirty-five IgM-positive and 57 IgM-negative human sera detected by the imported IgM-ELISA kit were detected with the rSAG1-IgM-ELISA. Results The purity of rSAG1 was above 90%. The appropriate detecting condition was that the coated rSAG1 was 2 5 ?g/ml, the human serum was in 1∶100 dilution, and the second antibody was in 1∶4000 dilution. The coefficient of variation (CV) value of IgM-positive and IgM-negative pooled sera were 13 8% and 7 7% respectively. The inhibition rate of the assay was 62 0% The positive correspondence rate and negative correspondence rate were 82 9% (29/35) and 91 2% (52/57) respectively,the total correspondence rate was 88 0%, compared with the imported IgM-ELISA kit. Conclusions The rSAG1-IgM-ELISA has high sensitivity and specificity, and good correspondence rate with the imported IgM-ELISA kit. It indicates that rSAG1-IgM-ELISA has potential value for early diagnosis of toxoplasmosis.

Objective To evaluate the role of sclerostin in bone loss of postmenopausal Chinese women with type 2 diabetes mellitus.
Methods The postmenopausal patients suffering from type 2 diabetes mellitus and age, body mass index, and duration of menopause matched healthy controls were enrolled into this cross-sectional study according to criteria of inclusion and exclusion. The serum sclerostin level and bone mineral density of the anterior-posterior lumbar spine (L1-L4), femoral neck, and total hip were determined by using a quantitative sandwich ELISA kit and dual X-ray absorptiometry, respectively. Meanwhile, the clinical and laboratory indexes of bone mineral metabolism were analyzed. Associations between serum sclerostin level and bone mineral density as well as bone turnover markers were evaluated by linear regression analysis.
Results Finally, 265 postmenopausal women with type 2 diabetes and 225 non-diabetic women were recruited in the diabetic group and control group, respectively. Serum sclerostin level of the diabetic group was significantly higher than that of the control group (48.2±19.4 vs. 37.2±18.6 pmol/L, P<0.001) and was increased with age in both groups (diabetic group, r=0.374, P<0.001;control group, r=0.312, P<0.001). In type 2 diabetes patients, serum sclerostin concentration was positively correlated with hemoglobin A1c level (r=0.237; P=0.021). Biochemical bone turnover markers, intact parathyroid hormone and bone-specific alkaline phosphatase, were negatively associated with serum sclerostin level (r=?0.138, P=0.078 and r=?0.265, P<0.001). Conversely, the positive correlation between sclerostin and C-terminal cross-linking telopeptide of type I collagen was found in diabetic patients (r=0.354, P<0.001). Serum sclerostin levels of the diabetic group were positively correlated with bone mineral density of the lumbar spine, femoral neck, and total hip (r=0.324, 0.367, and 0.416, respectively;all P<0.001).
Conclusions Sclerostin might participate in the pathogenesis of bone loss of type 2 diabetes. The high sclerostin level might serve as a marker of increased osteocyte activity in postmenopausal patients with type 2 diabetes mellitus.

Objective To investigate the potential role of nucleotide-binding oligomerization domain 1 (NOD1), a component of the innate immune system, in mediating lipid-induced insulin resistance in adipocytes.
Methods Adipocytes from Toll-like receptor 4 deficiency mice were used for stimulation experiments. The effect of oleate/palmitate mixture on nuclear factor-κB (NF-κB) activation was analyzed by reporter plasmid assay. The release of proinflammatory chemokine/cytokines production was determined by using real-time PCR. Insulin-stimulated glucose uptake was measured by 2-deoxy-D-[3H] glucose uptake assay. Chemokine/cytokine expression and glucose uptake in adipocytes transfected with small interfering RNA (siRNA) targeting NOD1 upon fatty acids treatment were analyzed.
Results Oleate/palmitate mixture activated the NF-κB pathway and induced interleukin-6, tumor necrosis factor-α, and monocyte chemoattractant protein-1 mRNA expressions in adipocytes from mice deficient in Toll-like receptor 4, and these effects were blocked by siRNA targeting NOD1. Furthermore, saturated fatty acids decreased the ability of insulin-stimulated glucose uptake. Importantly, siRNA targeting NOD1 partially reversed saturated fatty acid-induced suppression of insulin-induced glucose uptake.
Conclusion NOD1 might play an important role in saturated fatty acid-induced insulin resistance in adipocytes, suggesting a mechanism by which reduced NOD1 activity confers beneficial effects on insulin action.

This study was aimed to establish the quantitative analysis of hIL-2 in culture supernatant by multifunctional Luminex 100. The lymphocytes were separated from ACD-anticoagulated peripheral blood by density gradient method. The lymphocytes were stimulated with PHA for 48 hours, and frozen at -20 degrees C The relative fluorescence units of standard preparations and samples were detected by multifunctional Luminex 100, and the sample concentrations were calculated by standard curve. The results indicated that the regression equation of standard preparation is Lg (RFU) = 1.547 + 0.867 LgC. ANOVA F = 301.7427, p < 0.05 (nu = 6). The analysis of variance showed F = 301.7427, p < 0.05 (nu = 6). The test of regression coefficient showed t = 17.3707 (nu = 6), p < 0.05. It is concluded that method for induction and measurement of human IL-2 in vitro is established. The standard curve established by this way is statistically significant. There is linear relationship between the concentration of hIL-2 and fluorescence intensity.
Cell Separation ; methods ; Humans ; Interleukin-2 ; analysis ; Lymphocytes ; cytology ; drug effects ; metabolism ; Phytohemagglutinins ; pharmacology

OBJECTIVETo investigate the effect of chemokine stromal cell-derived factor-1 (SDF-1) and its receptor CXCR4 on liver metastasis of human colon cancer.

METHODSExpression of CXCR4 in different colon cancer cell lines and SDF-1 in different tissues were detected by using Western-blot technique. Effect of SDF-1 and anti-CXCR4 monoclonal antibody (McAb) on proliferation and migration of HT-29 cells were measured using MTT methods. Model mimicking liver metastasis of human colon cancer was established by injecting HT-29 cells intrasplenically into BALB/C nude mice. Mice were randomly divided into AMD3100 treated group and control group. Liver metastatic rate and tumor foci were measured 7 weeks after.

RESULTSHT-29 cells expressed higher level of CXCR4 protein, and liver tissue expressed higher level of SDF-1 protein. Compared with the control, SDF-1 could significantly induced the proliferation and migration of the HT-29 cells, and anti-CXCR4 McAb could inhibited both functions of SDF-1. The liver metastasis rate in the control group was 100%, and it was 40% in the AMD3100 treating group (P < 0.05). The mean liver metastasis number also significantly decreased by AMD3100 (7.8 +/- 2.6 vs 22.4 +/- 8.6, P < 0.05).

CONCLUSIONSSDF-1/CXCR4 biological axis play an important role in liver metastasis of human colon cancer. Arrest of CXCR4 can inhibit liver metastasis of colon cancer through blocking cell proliferation and migration induced by SDF-1.

Animals ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Chemokine CXCL12 ; metabolism ; physiology ; Colonic Neoplasms ; metabolism ; pathology ; HT29 Cells ; Humans ; Liver Neoplasms, Experimental ; secondary ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Receptors, CXCR4 ; metabolism ; physiology ; Xenograft Model Antitumor Assays

OBJECTIVETo evaluate the role of sclerostin in bone loss of postmenopausal Chinese women with type 2 diabetes mellitus.

METHODSThe postmenopausal patients suffering from type 2 diabetes mellitus and age, body mass index, and duration of menopause matched healthy controls were enrolled into this cross-sectional study according to criteria of inclusion and exclusion. The serum sclerostin level and bone mineral density of the anterior-posterior lumbar spine (L1-L4), femoral neck, and total hip were determined by using a quantitative sandwich ELISA kit and dual X-ray absorptiometry, respectively. Meanwhile, the clinical and laboratory indexes of bone mineral metabolism were analyzed. Associations between serum sclerostin level and bone mineral density as well as bone turnover markers were evaluated by linear regression analysis.

RESULTSFinally, 265 postmenopausal women with type 2 diabetes and 225 non-diabetic women were recruited in the diabetic group and control group, respectively. Serum sclerostin level of the diabetic group was significantly higher than that of the control group (48.2±19.4 vs. 37.2±18.6 pmol/L, P<0.001) and was increased with age in both groups (diabetic group, r=0.374, P<0.001; control group, r=0.312, P<0.001). In type 2 diabetes patients, serum sclerostin concentration was positively correlated with hemoglobin A1c level (r=0.237; P=0.021). Biochemical bone turnover markers, intact parathyroid hormone and bone-specific alkaline phosphatase, were negatively associated with serum sclerostin level (r=-0.138, P=0.078 and r=-0.265, P<0.001). Conversely, the positive correlation between sclerostin and C-terminal cross-linking telopeptide of type I collagen was found in diabetic patients (r=0.354, P<0.001). Serum sclerostin levels of the diabetic group were positively correlated with bone mineral density of the lumbar spine, femoral neck, and total hip (r=0.324, 0.367, and 0.416, respectively; all P<0.001).

CONCLUSIONSSclerostin might participate in the pathogenesis of bone loss of type 2 diabetes. The high sclerostin level might serve as a marker of increased osteocyte activity in postmenopausal patients with type 2 diabetes mellitus.

Aged ; Alkaline Phosphatase ; blood ; Asian Continental Ancestry Group ; Biomarkers ; blood ; Bone Morphogenetic Proteins ; blood ; China ; epidemiology ; Diabetes Mellitus, Type 2 ; blood ; epidemiology ; Female ; Genetic Markers ; Hemoglobin A ; metabolism ; Humans ; Middle Aged ; Osteocytes ; metabolism ; pathology ; Osteoporosis, Postmenopausal ; blood ; epidemiology ; Parathyroid Hormone ; blood ; Retrospective Studies

Objective To retrospectively study revisions for infected total hip replacements in 30 cases and discuss the bacteriological characters of the infected total hip replacements,difficulties and strategies in the revision.Methods Thirty revisions of infected total hip replacements were reviewed retrospectively.There were 12 males and 18 females,with mean age of 62.5 years(31-86 years)at revi- sion surgery.Infection was presented one month to four years(mean seven months)after THA operation. The diseases for initial operation included femoral neck fractures in 12 cases,femoral head necrosis in 11,hip osteoarthritis in five and rheumatoid arthritis in two.Twelve eases were treated by one-stage revi- sion and 18 by two-stage revision.Results Before the revision operation,the hip infection were diag- nosed by bacterial culture in 18 cases including five with Staphylococcus epidermidis,four with Staphylo- coccus aureus and nine with other bacteria.Bacteria growth appeared on the specimens from 23 hip joints during the revision surgery but not on the specimens from seven hip joints.Of 12 one-stage revisions,10 cases were followed for mean 16 months,which showed infection recurrence in two eases.Of 18 two-stage revisions,13 cases were followed for mean 20 months,which showed one case with infection recurrence. The mean Harris hip score was improved from preoperative 44 to 84 at follow-up.Conclusions 1) The main bacteria in the infected hip are antibiotic resistant Staphylococcus.2)Because the revision op- eration is difficult,careful preparation before revision is important for success.The fresh surgeon should not attempt.3)The revision strategies should vary according to specific status of the cases.The infection recurrence rate is lower when using a two-stage revision strategy.4)Application of antibiotic bone cement can help improve treatment effect and facilitate functional recovery of the joints.5)The scientific rehabil- itation after operation is very important to functional recovery.

Objective To report the experience of the application of microsurgery in joint replace- ment.Methods There were 22 cases,10 cases with segmental acetabular defects treated with the pedicle sartorius muscle iliac bone grafting,5 cases with vascular repair following major vascular injury of extremity during operation,6 cases with neural repair following neural injury during operation,1 case with serious injury reconstruction by elbow joint replacement and free flap.Results The operations succeeded in 22 cases without any postoperative infection.The mean follow-up was 40.1 months (3-72 months) in 22 cases,in which the joint function improved and the operative result was satisfactory with no joint pain.Conclusion Microsurgical technique can reconstruct bone and tissue defect effectively in joint replacement.

Objective:To investigate the relationship between oral health and mild cognitive impairment(MCI)by using indicators that evaluate oral hypofunction.Methods:The study was conducted using a cross-sectional design.Participants were recruited from three communities(Shizi Ling, Wenyi Xincun, and Yaoling in Furong District)in Changsha in July 2021, using convenience sampling.Cognitive and oral functions were evaluated using the simple mental state examination, version 2(MMSE-2)and seven indicators of oral hypofunction, which included oral hygiene, oral dryness, occlusal force, tongue and lip movement, tongue pressure, mastication, and swallowing function.Results:A total of 144 subjects were included in this study, with 72 males.Except for education level, there were no statistically significant differences between the MCI group and the normal cognitive group in terms of demographic information and self-reported and measured oral functions(all P>0.05).There was a correlation between the subjects' self-reported oral function and the actual measured oral function.Multiple linear regression analysis revealed no significant correlation between the MMSE score and the seven indicators used to measure oral hypofunction(all P>0.05).The MMSE score of the female group showed a negative correlation with mastication( β=-0.003, P=0.043), while the MMSE score of subjects with elementary school education also showed a negative correlation with mastication( β=-0.022, P=0.016).Additionally, the MMSE score of subjects with middle school education showed a positive correlation with the number of residual teeth( β=0.090, P=0.030). Conclusions:The self-reported oral function can serve as an initial assessment of overall oral function.However, among elderly individuals who do not show significant decline in oral function, there was no significant correlation between oral function and MCI.To accurately identify individuals with MCI, a more detailed sub-analysis with a larger sample size is required.