[Effect of methyl eugenol on nasal mucosal aquaporin 5 in rats with allergic rhinitis].

OBJECTIVE: To observe the effect of methyl eugenol on the expression of aquaporin (AQP) 5 in nasal mucosa of rats with allergic rhinitis and to explore its significance. METHODS: In the study, 128 Wistar rats were randomly divided into normal control group, AR model control group, budesonide positive control group, 80 mg/kg group, 40 mg/kg group, 20 mg/kg group and 10 mg/kg group, and ovalbumin (OVA) was used to establish the model of allergic rhinitis. After successful modeling, castor oil, budesonide and corresponding doses of methyl eugenol were given respectively. After 1, 2 and 4 weeks of administration, the distribution of AQP5 in nasal mucosa was observed by immunohistochemistry. The expression of AQP5 in nasal mucosa of each group was compared by Western blotting. The expression of AQP5 mRNA was compared with real-time PCR. RESULTS: AQP5 was mainly located in the glandular epithelium and ductal epithelial cell membrane and cytoplasm. The expression of AQP5 and AQP5 mRNA in nasal mucosa of the rats in the model control group was lower than that in the normal control group (P<0.05). AQP5 and AQP5 mRNA in nasal mucosa of the rats in each treatment group were higher than those in the model control group in varying degrees. The expression of AQP5 in the budesonide group was not significantly different from that in the normal control group 1, 2 and 4 weeks after drug intervention (P>0.05), but there was significant difference between the budesonide group and the model control group (P<0.05). The expression of AQP5 mRNA in the budesonide group was significantly different from that in the normal control group and the model control group (P<0.05).After 2 weeks of intervention, the expression of AQP5 in each dose group of methyleugenol was not significantly different from that in the budesonide group (P>0.05). After 1 week of intervention, there was no significant difference in AQP5 mRNA between the 20 mg/kg group and the normal control group (P>0.05), but there was significant difference between the 20 mg/kg group and the model control group (P<0.05). CONCLUSION: Methyl eugenol may increase the degree of edema of the nasal mucosa by reducing the expression of AQP5 and reduce the secretion of glands, thus alleviating the symptoms of allergic rhinitis, sneezing and runny nose.

[Effect of long-term deep slow-wave sleep deprivation on the reproductive system in male rats].

Objective: To investigate the effect of long-term deep slow-wave sleep deprivation on the gonad axis, sperm abnormality rate, and structure of the testis in male rats and possible mechanisms. Methods: A total of 30 specific pathogen-free male Wistar rats aged 5 weeks were randomly divided into slow-wave sleep deprivation group 1 (SD1 group) , slow-wave sleep and sleep time deprivation group 2 (SD2 group) , and control group, with 10 rats in each group. The flower pot method was used to establish a model of sleep deprivation. In addition to 12-hour sleep deprivation at night, the rats in the SD1 group were given interference once every 24 minutes, and those in the SD2 group were deprived of sleep for 8 minutes every 24 minutes; the rats in the control group were given 12-hour light illumination and then placed in dark environment for 12 hours. All rats were sacrificed by exsanguination from the femoral artery, and the testis, the epididymis, and blood were collected for analysis. Sperm abnormality rate and sperm motility rate were measured, and cauda epididymal sperm counting was performed. ELISA was used to measure the serum levels of testosterone (T) , follicle-stimulating hormone (FSH) , and luteinizing hormone (LH) . Results: Compared with the control group, the SD2 group had a significant increase in organ coefficient of the epididymis (P<0.05) and a significant reduction in sperm motility rate (P<0.05) . There were significant differences between the SD1 group and the SD2 group in the increase in sperm abnormality rate (P<0.05) and the reduction in cauda epididymal sperm count (P<0.05) . The levels of FSH and T tended to increase, and the level of LH tended to decrease. Pathological examination showed degeneration and vacuolization of a small amount of spermatogenic cells in the SD1 group; in the SD2 group, there were significant degeneration, edema, and vacuolization of most spermatogenic cells, some spermatogenic cells were observed in the lumen, and there were no sperms in the lumen. Conclusion: Long-term deep slow-wave sleep deprivation impairs the structure of the testis, affects sperm motility rate and sex hormones, and increases the risk of sperm abnormality.

Effects of hTERT antisense oligodeoxynucleotide on cell apoptosis and expression of hTERT and bcl-2 mRNA in keloid fibroblasts.

OBJECTIVE: This study was purposed to investigate the effects of hTERT antisense oligodeoxynucleotide (ASODN) on cell apoptosis and expression of hTERT and bcl-2 mRNA in keloid fibroblasts and to explore its anti-keloid effect. MATERIALS AND METHODS: Primary cultures of dermal fibroblasts derived from 12 keloid samples were established, strains of fibroblasts at passages 3 to 4 were used in this study. After treated by hTERT ASODN the proliferation of the fibroblasts was measured by cell count and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay method, the apoptosis was analyzed by flow cytometry (FCM), and the expression of hTERT and bcl-2 mRNA were observed by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). The data was analyzed by statistical software (SPSS11.5). RESULTS: The results showed that after sealing hTERT gene with ASODN for 72 h, the fibroblasts growth was repressed and the ability of proliferation decreased as the fibroblasts were treated with 1.0 mol/L ASODN for 72 h, the fibroblasts apoptosis was induced and the expression of hTERT and bcl-2 mRNA was lower than that of controlled group. The result was significantly different between control group and treatment group and was related to the treatment time of ASODN (p<0.01), but the difference was no significant when compared 1.0 µmol/L SODN group with untreated group (p>0.05). CONCLUSIONS: As a negative modutory factor, hTERT-ASODN can suppress growth and proliferation of keloid fibroblasts. Decreasing the telomerase activity of keloid fibroblasts may be one of the most important mechanisms. That hTERT-ASODN inhibited telomerase activity in keloid fibroblasts is an important pathway that may play a key role in the anti- keloid therapy.

The expression of inhibitor of growth 4 is reduced in cervical cancer tissues.

OBJECTIVE: Inhibitor of growth 4 (ING4) is a candidate tumor suppressor which plays an important role in multiple processes including DNA repair, apoptosis, cell cycle control, tumor metastasis and angiogenesis. However, clinical data about the role of ING4 in the development and progression of cervical cancer are still limited. This study aimed to examine ING4 expression in cervical cancer and analyze its correlation with the progression of the malignancy. PATIENTS AND METHODS: RT-PCR, Western blot and immunohistochemistry analysis were performed to determine ING4 expression in 18 clinical specimens from cervical cancer patients. The correlation of ING4 expression with the clinical-pathological features of the patients was analyzed. Moreover, the correlation between ING4 and HPV E6/E7 transcription level in SiHa cells was analyzed. RESULTS: ING4 expression was decreased significantly at mRNA and protein levels in the tissues of cervical cancer compared with paracarcinoma tissues. Analysis of the subcellular localization of ING4 showed that ING4 expression was decreased in the nucleus of cervical cancer tissues. Ectopic expression of ING4 reduced the proliferation of SiHa cells, accompanied by decreased HPV E6/E7 transcription. CONCLUSIONS: ING4 expression is decreased in human cervical cancer tissues. Reconstitution of ING4 expression in cervical cancer cells is correlated with decreased HPV E6/E7 transcription. These data suggest that ING4 expression has diagnostic and prognostic significance for cervical cancer.

Size-dependent oriented attachment in the growth of pure and defect-free hexagonal boron nitride nanocrystals.

Pure and defect-free hexagonal boron nitride (hBN) nanocrystals with deep-ultraviolet light emissions at around 215 nm were prepared via a solid state reaction. This involved preparing a precursor from potassium borohydride and ammonium chloride powders, and then heating the precursor and additional ammonium chloride to 1000 °C within a nitrogen atmosphere. The hBN nanocrystals were studied using a variety of characterization techniques (e.g., TEM, AFM, N(2) absorption/desorption). A growth mechanism based on size-dependent oriented attachment was proposed for the nanocrystals.

The impact of iodine excess on thyroid hormone biosynthesis and metabolism in rats.

Thyroid function ultimately depends on appropriate iodine supply to the gland. There is a complex series of checks and balances that the thyroid uses to control the orderly utilization of iodine for hormone synthesis. The aim of our study is to evaluate the mechanism underlying the effect of iodine excess on thyroid hormone metabolism. Based on the successful establishment of animal models of normal-iodine (NI) and different degrees of high-iodine (HI) intake in Wistar rats, the content of monoiodotyrosine (MIT), diiodotyrosine (DIT), T(4), and T(3) in thyroid tissues, the activity of thyroidal type 1 deiodinase (D1) and its (Dio1) mRNA expression level were measured. Results showed that, in the case of iodine excess, the biosynthesis of both MIT and DIT, especially DIT, was increased. There was an obvious tendency of decreasing in MIT/DIT ratio with increased doses of iodine intake. In addition, iodine excess greatly inhibited thyroidal D1 activity and mRNA expression. T(3) was greatly lower in the HI group, while there was no significant difference of T(4) compared with NI group. The T(3)/T(4) ratio was decreased in HI groups, antiparalleled with increased doses of iodine intakes. In conclusion, the increased biosyntheses of DIT relative to MIT and the inhibition of thyroidal Dio1 mRNA expression and D1 activity may be taken as an effective way to protect an organism from impairment caused by too much T(3). These observations provide new insights into the cellular regulation mechanism of thyroid hormones under physiological and pathological conditions.

Retrospective analysis of thyroid nodules by clinical and pathological characteristics, and ultrasonographically detected calcification correlated to thyroid carcinoma in South China.

OBJECTIVE: To investigate the clinical and pathological characteristics of thyroid nodules, as well as to evaluate the significance of ultrasonographically detected thyroid calcification in the diagnosis of thyroid carcinomas. METHODS: Retrospective data were studied from 1,051 consecutive patients who underwent a thyroidectomy in the Provincial Hospital of Fujian Medical University in South China between January 2003 and July 2006 for nodular thyroid disease. Complete sonographical information before surgery was only collected from 758 of the 1,051 patients. RESULTS: Among the 1,051 patients, benign lesions were found in 857 (81.54%) patients, of whom 612 (71.41%) were nodular goiter; malignant lesions were found in 194 (18.46%) patients, in whom benign thyroid lesions were also found in 85 (43.81%) patients. A total of 48 patients suffered from microcarcinomas, of whom 37 patients had benign lesions; these 37 accounted for 43.53 and 77.08%, respectively, of the 85 malignant cases with benign lesions and the 48 cases with microcarcinomas. In the 758 patients who underwent thyroid ultrasonography before surgery, intrathyroidal calcifications were apparent in 243 patients (32.06%). The incidence of calcification was significantly higher in patients with thyroid carcinoma (54.17%) than in those with benign lesions (26.87%; p < 0.005). Detection of calcification in thyroid lesions by ultrasound had a sensitivity of 32.38% and a specificity of 87.35%, with an OR of 3.31 (95% CI, 2.24-4.63), positive likelihood ratio of 2.56, negative likelihood ratio of 0.77 and a kappa value of 0.23. CONCLUSION: Thyroid carcinoma, especially microcarcinoma, often coexists with benign thyroid disease. Calcification detected by thyroid ultrasound represents a risk factor for malignancy, but is of limited use as a sole marker of malignancy.

[Effect of uncaria on evoked field potential of hippocampal slice in epileptogenic rat].

AIM: To investigate the effect of uncaria on evoked field potential of CA1 hippocampal slice in epileptogenic model. METHODS: We made rats which were injected by pilocarpine as epileptic-models. The pathway of drug administration is titration adjacent to hippocampal slices. We recorded the evoked field potential of hippocampal slice by glass microelectrode extracellularly, and observed the effect of uncaria on the amplitude of population spike (PS) in CA1 pyramidal cells in hippocampal slices. RESULTS: The uncaria with concentration of 1 g/ml could reduce the amplitude of PS in CA1 of hippocampal slices. The average of fall was 27.64%, and restored in 8.71 minutes on average (n = 14, P < 0.01). CONCLUSION: Uncaria could reduce the amplitude of PS in CA1 of epileptic hippocampal slices remarkably. These results show that uncaria can inhabited the synaptic transfer activity of CNS remarkably, suggest that it has the distinct effect of anti-epilepsy.

Differential binding of (+) and (-) gossypol to plasma protein and their entry into rat testis.

Concentrations of (+) and (-) gossypol were measured by high performance liquid chromatography after they were incubated with plasma proteins in vitro. The concentration of (-) gossypol decreased more than the concentration of (+) gossypol. A similar decrease in free gossypol concentrations in the blood plasma of rats was observed after intravenous infusion of gossypol enantiomers. The concentration of (-) gossypol was also found to be lower than the concentration of (+) gossypol at the blood-testis barrier. The biological effect of (-) gossypol probably results from its stereospecific binding to extra- and intracellular proteins in vivo and inhibition of the biological activity of some proteins.

Immunization strategies for the production of rat monoclonal anti-idiotope antibodies.

Anti-idiotypic antibodies are powerful reagents for the study of immunoregulation, and have potential interest as vaccines against tumors and infectious diseases. Three immunization strategies for the production of rat monoclonal anti-idiotope antibodies have been compared in this paper. Male Wistar rats were immunized i.p. and at multiple subcutaneous sites with 750 micrograms of purified monoclonal antibody against Plasmodium falciparum for three times and subsequently boosted by (1) intraperitoneal injection with 750 micrograms of the immunogen, (2) intravenous inoculation with 400 micrograms of the IgG, and (3) intrasplenic immunization with 200 micrograms of the idiotype. With the intraperitoneal boost method, the frequency of hybrids with anti-idiotope activity was 0.3-0.9% with 62.8-85.2% of the seeded wells containing hybrids. In the intravenous boost group, the percentage of hybrids demonstrating anti-idiotope activity increased to 11.0-13.3% with 80.2-97.9% of the hybrid efficiency. When immunized by the intrasplenic boost route, the frequency of anti-idiotope hybrids generated rose to 12.9-16.4% with 82.3-96.6% of the hybrid efficiency. There was no obvious effect of the boost immunizing methods on the generation of rat monoclonal anti-mouse IgG antibodies. These results indicated that the multiple-site immunization followed by intravenous or intrasplenic boost injection was an appropriate immunizing method for the production of monoclonal anti-idiotope antibodies.

Cholesterol requirement for growth of IR983F and P3X63-Ag8-U1 myeloma cells in serum-free medium.

Cholesterol, a major lipid component of the plasma membrane, is thought to have profound effects on the structure and function of cells. Most animal tissues are capable of synthesizing cholesterol de novo from acetate; however, there are relatively few mammalian cells in vitro expressing an absolute requirement for an exogenous source of cholesterol. In this paper, it was shown that both IR983F (983) rat myeloma cells and P3X63-Ag8-U1 (P3U1) mouse myeloma cells which had been cultivated in serum-free medium containing cholesterol for more than 6 months still required cholesterol in vitro for growth in serum-free medium. Optimal growth of 983 and P3U1 occurred in cholesterol concentrations of 15 and 5 micrograms/ml, respectively. Moreover, it was demonstrated that the cholesterol could be replaced by human low density lipoprotein in a concentration of 10 micrograms/ml but not by mevalonic acid lactone. In contrast to the parental myeloma cells, hybridoma cells derived from the mouse myeloma cells which had been cultivated in serum-free medium containing cholesterol for more than 6 months did not require cholesterol.