Animals ; Calcium ; metabolism ; Cell Line ; Cricetinae ; Cricetulus ; Dose-Response Relationship, Drug ; Fibroblasts ; drug effects ; metabolism ; Vehicle Emissions

Objective To study the effects of selenium deficiency,iodine deficiency and combined selenium and iodine deficiency on bone and cartilage growth in the parental and the first filial generation rats. Methods Forty-eight weanling healthy SD rats were randomly divided into selenium deficieney, iodine deficiency, combined selenium and iodine deficiency and control groups according to their body mass. These rats were fed with selenium deficiency, iodine deficiency, combined selenium and iodine deficiency, and normal fodder, respectively. The parental rats (about 3 months old) were mated in each group 8 weeks after the beginning of the experiment. Right tibias and left knee joints were collected when the parental generation rats were about 6 months and the first filial generation rats were about 3 months old. Tibial length, mid-shaft tibial diameter, and articular cartilage diameters of the right tibias were measured by vernier caliper. Left knee joints were embedded and cut into sections and the thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes in growth plate cartilage were observed under the light microscope. Results The selenium deficiency had significant effect on serum selenium level of the parental and the first filial generation rats(F value were 239.56,232.68, P< 0.01), and also on serum T4 level of the first filial generation rats(F value were 6.95, P < 0.05). The iodine deficiency had significant effect on serum T3 and T4 level in the two generations rats(F value were 14.11,14.05,30.29,34.53, P < 0.01 ). There were interactions between selenium deficiency and iodine deficiency on serum T4 level in the first filial generation rats (F= 5.99, P< 0.05). The serum selenium of selenium deficiency group[ (30.28 ± 6.34), (43.95 ± 9.75)μg/L],combined selenium and iodine deficiency group[ (30.33 ± 5.18), (35.40 ± 3.16)μg/L] were significantly lower than iodine deficiency group[(345.83 ± 29.55), (245.24 ± 9.95)μg/L] and the controls[ (358.64 ± 30.50), (236.50 ±9.75) μg/L] in the two generations. The serum T3 of combined selenium and iodine deficiency group [(0.55 ± 0.05 ),(0.88 ± 0.14)nmol/L] were significantly lower than the controls[(0.75 ± 0.08), (1.26 ± 0.26)nmol/L] in the two generations. The serum T4 of iodine deficiency [ (24.11 ± 2.29), (42.10 ± 8.92) nmol/L ] and combined selenium and iodine deficiency group[ (20.66 ± 1.93), (26.55 ± 5.98)nmol/L] were significantly lower than the controls[ (36.15 ±2.74), (52.79 ± 8.84)nmol/L] and selenium deficiency group[ (28.12 ± 3.33), (52.02 ± ll.99)nmol/L] in the two generations. The selenium deficiency and iodine deficiency had significant effect on tibial length, thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes in first filial generation rats(F values were 24.31,6.98,40.76,56.15,25.24,82.82, 10.07,5.57, P <0.05 or <0.01). There were interactions between selenium deficiency and iodine deficiency on tibial length, thickness of the growth plate cartilage, layers of proliferative and hypertrophic chondrocytes (F values were 5.68,24.86,41.82,9.12, P <0.05 or <0.01 ). The tibial length of the selenium deficiency group[ (33.17 ± 0.34)mm] and combined selenium and iodine deficiency group[ (31.30 ± 0.87)mm] were significantly lower than the controls[ (34.12 ± 0.32)mm, P< 0.05]. Thickness of the growth plate cartilage [ (1.60 ± 0.18)mm ], layers of proliferative chondrocyte (8.54 ± 0.81), and hypertrophic chondrocyte (4.95 ± 0.37)of the combined selenium and iodine deficiency group were significantly decreased when compared to the selenium deficiency group[ (3.03 ± 0.10)mm, 14.68 ± 0.84,6.60 ± 0.31], iodine deficiency group[ (2.90 ± 0.09)mm, 13.75 ±0.33,6.61 ± 0.84 ] and the controls [ (3.19 ± 0.09) mm, 14.94 ± 0.36, 6.64 ± 0.26, P <0.05]. Thickness of the growth plate cartilage, layers of proliferative chondrocyte of the iodine deficiency group were lower than the controls(P<0.05). Conclusions Selenium deficiency impair tibial growth in first filial generation rats, iodine deficiency retarded the chondroncyte proliferation and decreases the thickness of growth plate cartilage in first filial generation rats, and combined selenium and iodine deficiency significantly impair the growth of bone and cartilage in first filial generation rats.

OBJECTIVETo study the impact of low-level lead exposure on neural cell adhesion molecule (NCAM) expression of primarily cultured hippocampal neurons.

METHODSWistar rats gestated at 18th day were anaesthetized and paunched to get the pups, the hippocampi of the pups were separated and the hippocampal neurons were primarily cultured. After co-cultivated with different dosage of PbCl(2), the NCAM expression of the neurons were tested with Western blotting at different culture time.

RESULTSNormally, the expression of NCAM at the 1st culture day was very low and its integral obsorbency density was 14; the climax expression time of NCAM of the cultured hippocampal neurons was 3rd to 5th cultured day, and their integral obsorbency density were 2 542 to 2 580; henceforth, the NCAM expression declined. NCAM expression was inhibited significantly by lead during the 2nd to 4th cultured day, and dose-response relationship was observed. The inhibition of lead weakened along with the cultured time prolonged, at 5th cultured day, it disappeared, and the NCAM expression of 10(-2), 10(-3) and 10(-4) mmol/L groups even exceeded the control groups. After that, the expression of NCAM in all groups began to decline, and the dose-response relationship of lead to the NCAM expression was observed again.

CONCLUSIONLow-level lead might significantly inhibit the NCAM expression of the primarily cultured Wistar rats' hippocampal neurons, and might delay the climax NCAM expression time.

Animals ; Animals, Newborn ; Cell Separation ; Cells, Cultured ; Dose-Response Relationship, Drug ; Female ; Hippocampus ; cytology ; drug effects ; metabolism ; Lead ; toxicity ; Neural Cell Adhesion Molecules ; biosynthesis ; genetics ; Neurons ; cytology ; Pregnancy ; Rats ; Rats, Wistar

OBJECTIVETo observe cell apoptosis and Bcl-2 and Bax expression changes of chondrocytes induced by butenolide (BUT) and the inhibitory effect of selenium against BUT-induced chondrcyte apoptosis, to gain insights into the mechanism by which BUT induces chondrcyte apoptosis.

METHODSCartilage tissue reestablished from human fetal articular chondrocytes in vitro were treated with BUT at the concentrations of 0.1, 1.0 and 5.0 microg/ml and with the protective factor selenium. TUNEL method was used to detect chondrocyte apoptosis, which was quantified by flow cytometry. Immunohitochemistry was performed to analyze the expression of Bcl-2 and Bax in the reestablished cartilage tissue.

RESULTSBUT exposure induced chondrocyte apoptosis, and the apoptosis rate increased with the concentration increment of BUT from 0 to 1.0 mg/ml, resulting also increased positive expression rate of Bcl-2 and Bax(P<0.05). The apoptosis rate of chondrocytes in BUT+ selenium group was significantly lower than that of BUT groups (P<0.05), as was the positivity rate of Bcl-2 and Bax expression (P<0.05).

CONCLUSIONBUT induces chondrocyte apoptosis in positive relation with BUT concentration (from 0 to 1.0 mg/ml) and causes increased expressions of Bcl-2 and Bax. Selenium can inhibit the chondrocyte apoptosis induced by BUT.

4-Butyrolactone ; analogs & derivatives ; pharmacology ; Apoptosis ; drug effects ; Cells, Cultured ; Chondrocytes ; drug effects ; Humans ; In Situ Nick-End Labeling ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Selenium ; pharmacology ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo investigate chondrocyte apoptosis and expression of Fas and inducible nitric oxide synthase (iNOS) in articular cartilage in the pathogenesis of Kashin-beck disease (KBD) and primary osteoarthritis (OA).

METHODSThe collected samples of articular cartilage were divided into three groups: normal control (15 cases), KBD adults (15 cases) and OA (15 cases). Chondrocyte apoptosis was detected by terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling method, and Fas and iNOS in articular cartilage were stained by immunohistochemistry.

RESULTSThe positive percentages of chondrocyte apoptosis stained in articular cartilage of KBD and OA were significantly higher than that of the control (P < 0.01), and the positive percentage of chondrocytes apoptosis in the eroded areas of articular cartilage were significantly higher than in the non-eroded areas in articular cartilage of the same patient with KBD and OA (P < 0.05). There was no significant difference in positive percentage of chondrocytes apoptosis between KBD and OA. The positive percentages of Fas and iNOS in chondrocytes were significantly higher in KBD and OA than in control (P < 0.01). Significant differences in Fas and iNOS expression between the eroded areas and non-eroded areas were seen in articular cartilage of patients with KBD and OA (P < 0.05), but such difference did not exist between KBD and OA.

CONCLUSIONCell apoptosis seems to be associated with the pathogenesis of both KBD and OA. Fas and iNOS might mediate chondrocyte apoptosis.

Adult ; Apoptosis ; Cartilage, Articular ; pathology ; Chondrocytes ; cytology ; Endemic Diseases ; Female ; Humans ; In Situ Nick-End Labeling ; Male ; Nitric Oxide Synthase ; metabolism ; Osteoarthritis ; pathology ; physiopathology ; Osteoarthritis, Knee ; pathology ; physiopathology ; fas Receptor ; metabolism

OBJECTIVETo investigate chondrocyte apoptosis and the expressions of Bcl-2, Bax, Fas and iNOS in the articular cartilage between Kashin-Beck disease (KBD) and primary osteoarthritis (OA) and explore the difference in pathogenesis between the two diseases.

METHODSThe articular cartilage specimens were collected from 15 normal human subjects, 15 adult patients with KBD and 15 with OA. Chondrocyte apoptosis was detected by TUNEL method, and the expressions of Bcl-2, Bax, Fas and iNOS in articular cartilage were examined with B-SA immunohistochemistry.

RESULTSThe percentages of apoptotic chondrocytes positive for TUNEL staining in the articular cartilage were significantly higher in patients with KBD and OA than in normal control subjects (F=20.90-53.16, df=42, P<0.01), and the erosive areas of the articular cartilage contained greater percentage of apoptotic chondrocytes than the non-erosive areas in the same patient with KBD (t=4.154, df=28, P<0.01) or OA (t=6.004, df=28, P<0.01). No significant difference was noted in the positive apoptotic chondrocytes between KBD and OA (t=1.329-1.362, df=28, P>0.05). The percentage of chondrocytes positive for Bcl-2, Bax, Fas and iNOS were significantly higher in KBD and OA patients than in the control subjects (F=25.46-215.31, df=42, P<0.01), and significant differences were observed in Bcl-2, Bax, Fas and iNOS expressions between the erosed areas and non-erosed areas in articular cartilage in patients with KBD (t=2.608-7.77, df=28, P<0.05) and OA (t=2.278-5.413, df=28, P<0.05), but their expressions showed no significant difference between the two diseases (t=0.284-1.590, df=28, P>0.05).

CONCLUSIONThere was no significant difference in apoptotic chondrocytes and Bcl-2, Bax, Fas and iNOS expressions in the cartilage between adult patients with KBD and OA.

Adult ; Apoptosis ; Cartilage, Articular ; pathology ; Chondrocytes ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Osteoarthritis ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; bcl-2-Associated X Protein ; biosynthesis

OBJECTIVETo explore the effects of selenium and/or iodine deficiency on chondrocyte apoptosis in articular cartilage in rats.

METHODSForty-eight Sprague-Dawley rats were randomly divided into selenium deficiency group, iodine deficiency group, combined selenium and iodine deficiency group, and control group. Chondrocyte apoptosis was detected by terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) method, and Bcl-2 and Bax in articular cartilage were stained by immunohistochemistry in F3 generation of rats.

RESULTSIn articular cartilage, the positive rate of apoptotic chondrocytes stained by TUNEL in the upper and middle zones in selenium deficiency group, iodine deficiency group, and combined selenium and iodine deficiency group (all P < 0.05) were significantly higher than that in control group. The apoptotic chondrocytes were prominent in the middle zone. The positive percentage of chondrocytes apoptosis was not significantly different among these three groups (P > 0.05). Compared with the control group, the expressions of both Bcl-2 and Bax were significantly higher in the upper and middle zone in the selenium deficiency group, iodine deficiency group, and combined selenium and iodine deficiency group (all P < 0.05); however, the expressions of Bcl-2 and Bax were not significantly different among these three groups (P > 0.05).

CONCLUSIONSelenium and/or iodine deficiency may induce chondrocyte apoptosis.

Animals ; Apoptosis ; Cartilage, Articular ; metabolism ; pathology ; Chondrocytes ; metabolism ; pathology ; Female ; Iodine ; deficiency ; Male ; Rats ; Rats, Sprague-Dawley ; Selenium ; deficiency

Humans ; Uric Acid ; Sleep Apnea, Obstructive ; Risk Factors

Objective:To investigate the pharmacodynamics and mechanism of Chaijin Sanjie prescription (CJSJP) on rat mammary gland hyperplasia, in order to provide experimental basis for the research and development of new Chinese medicine. Method:SD rat model of mammary gland hyperplasia was established through exogenous intramuscular injection with estrogen and progesterone. After successful establishment of the model, the rats were randomly divided into normal group, model group, and low, medium and high-dose CJSJP groups (3.13, 6.26, 12.52 g·kg^-1) and Rupixiao (0.517 g·kg^-1) group, with 9 rats in each group. After 28 days of administration, estradiol (E₂), progesterone (P) and rolactin (PRL) were measured by radioimmunoassay, uterus and ovary coefficients were calculated; nipple diameter and breast histopathology were observed, estrogen receptor-α(ER-α) expression in mammary gland was measured by immunohistochemistry, and gonadotropin-releasing hormone (GnRH) and gonadotropin-releasing hormone receptor (GnRH-R) mRNA expressions in hypothalamus, pituitary were measured by Real-time PCR. ICR mice were randomly divided into normal group, low, medium and high-dose CJSJP groups (5.2,10.4,20.8 g·kg^-1) and Luotongding group (0.038 6 g·kg^-1) according to their body weight. Twelve mice in each group were given drugs for 7 days, and 0.6% acetic acid was injected intraperitoneally for 30 minutes after the last administration. The writhing times were observed within 15 minutes. Result:Compared with the normal group, the diameter of nipple was widened, serum E₂ was significantly increased (P<0.01),breast tissue proliferation and ER-α expression were increased in model group. compared with model group, the diameter of nipple was significantly decreased in high dose group of CJSJP (P<0.05), E₂ was decreased in all dose groups of CJSJP, pathological score of breast hyperplasia was decreased in middle and high dose groups of CJSJP, GnRH mRNA in hypothalamus was decreased in all dose groups of CJSJP. The writhing times of mice in high dose group of CJSJP was decreased (P<0.05). Conclusion:Chaijin Sanjie prescription can improve the lesions of breast hyperplasia. The therapeutic mechanism may be related to the regulation of GnRH gene expression in hypothalamus and the decrease of estrogen receptor expression.