Effect of selenium on apoptosis of spleen ceils, T-cell subset of fluorosis chicken / 中国地方病学杂志

Objective To study the effect of selenium on peripheral and splentic T-cell subset, apoptosis of spleen cells in fluorosis chicken and its mechanism. Methods One hundred and eighty 8-day Hailanhe chicks were randomly divided into 3 groups(each 60): ①control group: 195 mg/kg fluoride and 0.08 mg/kg of selenium; ②fluorine group : 1000 mg/kg fluoride and 0.08 mg/kg of selenium ;③selenium antagonism group : 1000 mg/kg On 30~(th), 60~(th), 90~(th) day, peripheral and splentic CD4~+, CD8~+ T-cell subset analyses underwent flow cytometry and apoptosis of spleen cells were detected by TUNEL for study subjects. Results Compared with control group, the CD4~+ T-cell subset of peripheral in fluorine group was decreased obviously in 30,60,90 days[ (35.36± 4.27)% vs (24.29 ± 2.96)%, (47.65 ± 5.42)% vs (41.62 ± 3.96)%, (49.58 ± 3.98) % vs (42.35 ± 6.03 )%, P < 0.05 or < 0.01 ], CD4~+/CD8~+ ratio also was decreased obviously [ ( 1.701 ± 0.145 )% vs (1.393 ± 0.163)%,(2.712 ± 0.345)% vs (1.781 ± 0.201)%,(2.438 ± 0.356)% vs (1.973 ± 0.229)%, P< 0.05 or < 0.01]. Compared with fluorine group, the CD4~+ T-cell subset of peripheral in selenium antagonism group [ (29.40 ± 3.38)%, (45.40 ± 6.01 )%, (46.85 ± 5.25)%, P < 0.05 or < 0.01 ] was increased obviously in 30,60,90 days,CD4~+/CD8~+ ratio in 60,90 days[(2.004 ±0.314)%,(2.211±0.229)%,all P<0.01]also was increased obviously.Compared with control group,the CD4~+ T-cell subset of spleen cells in fluorine group was decreased obviously in 30,60,90 days[(47.33±5.35)% vs(41.91±4.83)%,(49.28±5.24)% vs(41.26 ±4.56)%,(34.31±4.15)%vs(29.33±2.89)%,all P<0.01],CD4~+/CD8~+ ratio also was decreased obviously[(1.927 ±0.244)% vs(1.525 ±0.265)%,(1.847±0.224)% vs(1.640±0.198)%.(1.265±0.174)% vs(0.878±0.092)%,P<0.05 or<0.01].Compared with fluorine group,the CD4~+ T-cell subset of spleen cells in selenium antagonism group in 60,90 days[(44.87±5.43)%,(32.62±3.37)%,all P<0.05]was increased obviously,CD4~+/CD8~+ ratio in 30,60, 90 days[(1.703 ±0.201)%,(1.772±0.215)%,(0.991±0.124)%,P<0.05 or<0.01]also was increased obviously. The apoptosis ratio of spleen cells in fluorine group in 30,60,90 days[(2.31±0.36)%,(2.76±0.22)%,(3.04± 0.29)%]was higher than that in control group[(1.14±0.21)%,(1.23±0.23)%,(1.29±0.20)%,P<0.01].The apoptosis ratio of spleen cells in selenium antagonism group in 60,90 days[(2.42 ±0.32)%,(2.73±0.39)%]was lower than that in fluorine group(P<0.05 or<0.01).Conclusion A certain concentration of selenium can antagonize the immunity inhibition of fluorine by decreasing apoptosis and improving the unbalance of T-cell subset.

Effects of cold stress on energy metabolism in the chicken / 中国应用生理学杂志

<p><b>AIM</b>To investigate the effect of cold stress on the energy metabolism in Yisha chickens.</p><p><b>METHODS</b>Male Yisha chickens were subjected to acute (0.25, 1, 3, 6, 12 and 24 h) and chronic (5, 10 and 20 d) cold stress (12 +/- 1 degrees C). This study detected uncoupling protein (UCP) mRNA levels in gastrocnemius, glucagons (GLU) content in blood plasma and insulin (INS), blood glucose (BG) and free fatty acid (FFA) content in serum in the chicken.</p><p><b>RESULTS</b>The results were as follow: with the time lapsing during acute cold stress, UCP mRNA levels gradually increased, the content of INS and FFA showed fluctuant change, GLU content gradually increased, and BG content first increased and then decreased. During chronic cold stress, UCP mRNA levels significantly increased compared with their control group at every stress time point, and the content of INS, GLU, BG and FFA were all gradually increased with the time lapsing.</p><p><b>CONCLUSION</b>Cold stress could change the energy metabolism in chickens. And the different extent cold stress would produce different effects on the energy metabolism.</p>

Cloning the coding cDNA sequence of alpha1, beta2 and gamma2 subunit of GABA-A receptor in American king pigeon / 中国应用生理学杂志

<p><b>AIM</b>To clone and analyse the coding cDNA sequence of alpha1, beta2 and gamma2 subunit of GABAA receptor in American king Pigeon.</p><p><b>METHODS</b>Withdrew total RNA from the American king pigeon brain, reverse transcribing general primers to acquire a gene set cDNA. Designing specific primers of three subunit mRNA of the GABAA receptor, by RT-PCR respectively expanded the conservative gene of al subunit, beta2 subunit and gamma2 subunit of GABAA receptor, and carried on clone, plastid identification and the sequence measurese of three genes.</p><p><b>RESULTS</b>The experiment on sequence measures has succeeded that sequence analysis indicated that lengths of the conservative gene of alpha1 subunit, beta2 subunit and gamma2 subunit of GABAA receptor was respectively 899 bp, 597 bp and 563 bp, homology on reference sequence was respectively 94.99%, 94.64% and 96.28%.</p><p><b>CONCLUSION</b>Homology is high on the conservative gene of alpha1 subunit, beta2 subunit and gamma2 subunit of GABAA receptor of brain tissue of pigeon and chicken but there is a discriminating characteristic in different kinds of animals.</p>

Effect of fluoride on apoptosis and DNA damage in OX peripheral blood lymphocytes / 中国地方病学杂志

Objective To explore the effect of sodium fluoride on DNA damage and apoptosis on ox peripheral blood lymphocyte cultured in vitro.Methods Using lymphoeytes separation medium lymphocytes were separated and different concentration of NaF(0,4,8,12,16 mg/L)were added into the cultual system of lymphocytes for 24 h.Cell viability was measured by MTT.nuclear changes stained by acridine orange-ethidium bromine staining (AO/EB)were observed under fluoroscope,DNA fragment was measured by agarose gel electrophoresis.DNA damage was detected by alkaline SCGE.The differences between each groups were compared.Results Cells were exposed to 4,8,16 and 24 mg/L NaF in 24 h,cell survival rates,respectively being(73.743±0.552)%,(69.184±0.724)%,(65.736±0.055)%and(63.651±0.287)%,decreased significantly compared to control group.There were distinctive cell apoptosis,evident DNA damage and visible dose-effect relation(R2=0.7456).Conclusions A certain concentration of sodium fluoride lcads to lymphocytes apoptosis and DNA damage.

Effect of arsenic trioxide on apoptosis and mitochondrial transmembrane potential of Marek disease cancer cells / 中国地方病学杂志

Objective To investigate the mechanism of apoptosis induced by arsenic trioxide(As2O3) on MDCC-MSB1 cancer cell line in vitro. Methods MDCC-MSB1 cells were divided into 4 groups, treated with 0 (control group), 2, 4 or 8 μmoL/L of As2O3. At 48 h following the treatment, MTr assay was applied to detect the inhibitory effect of As2O3 on MDCC-MSB1. Morphological changes of apoptosis were observed by fluorescence microscopy. Apoptosis was examined by DNA Ladder. Changes of mitochondrial transmembrane potential were examined by Rhodamin 123 dye and flow cytometry. Results Inhibition ratio was 0, (5.34±3.02)%, (10.78± 0.55)% and (20.02±3.24)% respectively, along with the dosages of As2O3, the differences between the groups were statistically significant(P<0.01). Morphologic changes of apoptosis were observed by DNA ladder of agarose gel electrophoresis. Apoptosis rates were significandy increased from 3.200±0.459, 11.543±0.391, 17.206±0.636 to 21.343±0.620, and the differences between the groups were statistically significant(P<0.01). DNA ladder of experimental group was detected, Intact cell membrane, and mitochondrial transmembrane potential Pl-/Rh123- decreased apoptotic cells percentage was significantly increased from (1.06±0.14)%, (4.63±0.04)%, (9.62±0.07)% to (10.39±0.10)%, respective to doses of 0, 2,4, and 8 μmol of As2O3. The differences between the groups were statistically significant(P<0.01). Conclusions As2O3 can induce apoptosis in MDCC-MSB1 cells by decreasing the mitochondrial transmembrane potential.