Objective To discuss whether Omega-3 fish oil fat emulsion has the potential protective mechanism for 7-day-old rats with hypoxic-ischemic brain damage (HIBD).Methods One hundred and sixty-eight 7-day-old SD rats were randomly divided into 4 groups:group A (sham group),group B (Omega-3 fish oil fat emulsion group),group C (normal fat emulsion group),group D (model group),and there were 42 cases in each group.Neonatal HIBD was induced by the method of Rice.Rats were sacrificed at 1 d,3 d and 7 d after the surgery.Hippocampus was removed for Real-time PCR and Western blot test to detect Toll-like receptor 4 (TLR4) and nuclear factor-κB (NF-κB) mRNA and protein expression.TUNEL staining comparison was done among different groups to observe the number of cellular apoptosis.Results HE staining of hippocampus CA1 area in 3 d showed that brain tissues in group A maintained normal structures;those in group D had much more brain cells with severe edema than other groups;TLR4 and NF-κB mRNA and protein expression levels in group D were higher than those in group A in 1 d (all P ＜0.05);TLR4 and NF-κB expression levels of mRNA and protein in group B (4.89 ± 0.51,9.30 ± 1.53;1.15 ±0.10,1.44 ± 0.14) were lower than those in group C (17.58 ± 2.50,20.13 ± 1.00;2.56 ± 0.10,2.82 ± 0.09) and group D (15.94-± 2.52,26.21 ± 3.00;2.34 ± 0.11,4.51 ± 0.36) in 3 d (all P ＜ 0.05),and compared with group A (6.30 ± 1.52,5.32 ± 1.06;1.32 ± 0.10,2.42 ± 0.14),there was significant difference (all P ＞ 0.05);TLR4 and NF-κB mRNA and protein expression levels in group B were lower than those in group C and group D in 7 d(all P ＜0.05),and compared with group A there was no significant difference (all P ＞ 0.05).The apoptotic cell number of brain tissues in 3 d:group B (13.67 ±2.52) were lower than those in group C (27.67 ±2.52) and group D (41.00 ±3.61) (all P ＜0.05),and compared with the group A (6.00 ±2.00),the difference was not statistically significant (P ＞ 0.05).Conclusions Omega-3 fish oil fat emulsion plays an important role in protecting neonatal rats with HIBD.The mechanisms were likely to reduce TLR4,NF-κB and cell apoptosis levels.

Objective To investigate the expression of nuclear factor-kappa Bp65 (NF-κBp65)and Toll-like receptor 4(TLR4)protein in the brain tissues of 7-day-old Sprague-Dawley(SD) rats with cerebral hypoxia-ischemia encephalopathy (HIE) and to explore the role of TLR4 and NF-κBp65 in the pathogenesis of neonatal rats with hypoxic-ischemic brain damage.Methods Seven-day SD rats were randomly divided into the experimental group and the control group.Brain pathological changes were observed in light microscopy at 6 h、12 h、24 h、72 h、7 d after HIE.The expression of TLR4 and NF-κBp65 in brain tissues were analyzed by immunohistochemistry method.Results NF-κBp65 and TLR4 were expressed in the neuron and microglia of control group and experimental group.The expression were most significant at cerebral cortex and hippocamp.However,the expression of NF-κBp65and TLR4 began to increase at HIE 6h:NF-κBp65 (0.219 3 ± 0.024 7,0.215 7 ±0.030 4)and TLR4(0.327 1 ±0.033 3,0.303 9 ±0.037 9),and achieved the hightest at HIE 24h:NF-κBp65 (0.3564±0.0235,0.3365 ±0.023 2)and TLR4(0.434 2 ±0.0428,0.4193 ±0.041 3),then decreased at HIE 72 h:NF-κBp65 (0.289 2 ± 0.032 0,0.260 9 ± 0.021 2) and TLR4 (0.300 5 ± 0.020 9,0.282 0 ± 0.022 6),and HIE 7 d:NF-κBp65(0.247 9 ±0.0340,0.242 1 ±0.025 4) and TLR4(0.274 4 ±0.0288,0.257 1 ±0.027 5).Conclusion There is a positive correlation between NF-κBp65 and TLR4 in rats with HIE.It suggested that they may have the same pathophysiology development in HIE.

Objective To explore the effects of 75 mGy irradiation on the apoptosis of spermatogenic cells and antioxidant capacity of serum and testis and hormone levels in male rats with diabetes mellitus(DM).Methods Rats were injected intraperitoneally with streptozotocin(STZ)to develop diabetes.The diabetic rats were irradiated with 75 mGy X-rays every other day for 4 weeks.Their survival rate and body weight were recorded 12 weeks after development of diabetes.The apeptosis percentage of germ cells was measured with flow cytometry and TUNEL method.The changes of anti-oxidation and gonadal hormone levels in serum and testis were measured with kits.Results After the rats suffered from diabetes for 12 weeks,the survival rate in DM group was 25%(6/24),100% in normal control group(16116).The survival rate in 75 mGy + DM group(9/16,56.25%)was obviously higher than that in the DM group(X2= 4.00,P < 0.05).Meanwhile,the percentage of apaptotic spermatogenic cells in the diabetic rats was significantly larger than those in the normal control and irradiation groups(F = 5.496,P < 0.05).MDA and NO levels in serum and testis of diabetic rats were higher in varying degrees than that in the normal control,while the serum and testis MDA content in the 75 mGy + DM group were lower than those in the DM group especially in the testis(F = 10.644,P < 0.01).75 mGy X-ray irradiation decreased NO content in the diabetic rats serum significantly(F = 14.379,P < 0.05)and increased NOS activity and TS,FSH level(F = 9.676,43.194 and 5.282,respectively,P < 0.05 and P < 0.01).Conclusions LDR could decrease the MDA level and NO content,and increase the antioxidant enzyme activity and TS and FSH levels in testis and serum of diabetic rats.

Objective To construct the pshuttle-Egr-1-hSmac plasmid and transfect human breast cancer MDA-MB-435 cells,and to observe its radiotherapy enhancing effect on tumor cells.Methods The empty vector pshuttle and pshuttle-Egr-1-hSmac plasmid were transfected into MDA-MB-435 cells by liposomal.At different time(4,8,12,24 and 48 h)after irradiation with 2.0 Gy X-ray and 24 h after irradiation with 0.5 -5.0 Gy,the total RNA and protein were collected and extracted from these cells to analyze the Smac mRNA and protein expression levels with RT-PCR and Western blotting methods. The cells were divided into control, pshuttle, pshuttle-Egr-1-hSmac,2.0 Gy irradiation group, pshuttle + 2.0 Gy irradiation and pshuttle-Egr-1-hSmac+2.0 Gy irradiation groups.MTT method was used to evaluate cell proliferation,and the cell survival ability was measured with clone formation assay;Annexin Ⅴ/PI double staining and PI single staining were used to examine the apoptosis and cell cycle of MDA-MB-435 cells. Results There was no Smac mRNA expression in MDA-MB-435 cells in control and pshuttle groups,but the Smac mRNA expression levels in MDA-MB-435 cells in pshuttle-Egr-1-hSmac plasmid group were gradually increased with the time prolongation, and reached the maximum at 24 and 48 h;the Smac mRNA expression levels in MDA-MB-435 cells were increased gradually 24 h after irradiation of 0.5 - 5.0 Gy X-ray with the increasing of irradiation doses, and reached the maximum after 2.0 and 5.0 Gy irradiation. The Smac protein expression levels in pshuttle-Egr-1-hSmac plasmid group were increased gradually with the time prolongation,and reached the maximum at 24 h.The Smac protein expression lervels were increased 24 h afer irradiation of 0,0.5,1.0,2.0 and 5.0 Gy X-ray,especially in 5.0 Gy group. The MTT results showed that the A490 values in 2.0 Gy,pshuttle+2.0 Gy and pshuttle-Egr-1-hSmac groups 24, 48,and 72 h after irradiation were lower than those in control group(P<0.01);the A490 values of MDA-MB-435 cells in pshuttle-Egr-1-hSmac group after 1.0-5.0 Gy X-ray irradiation were lower than those in 0 Gy group (P<0.05 or P<0.01);the survival fraction(SF)in pshuttle-Egr-1-hSmac group was lower than those in control group (P<0.01).The percentages of the cells at G0/G1 and S phase in pshuttle-Egr-1-hSmac group were lower than those in 2.0 Gy group(P<0.01),the percentage of the cells at G2/M phase was higher than that in 2.0 Gy group (P<0.01);the apoptotic rate of the cells in pshuttle-Egr-1-hSmac group was higher than that in 2.0 Gy group (P<0.01).Conclusion X-ray irradiation can significantly increase the Smac mRNA and protein expression levels in MDA-MB-435 cells transfected with pshuttle-Egr-1-hSmac plasmid,inhibit the cell survival rate,and induce G2/M arrest and apoptotic increasing;Smac gene combined with radiotherapy could significantly increase the radiosensitivity of breast cancer cells.

This study was aimed to verify the effects of staged sequential therapy on expressions of matrix metalloproteinase-9 (MMP-9) and its inhibitor TIMP-1 within lung tissues in asthmatic rats with the airway remodeling, by applying a series of tests such as the immunohistochemistry, western blot and real-time fluorescent quantitative PCR. SD rats were randomly divided into 7 groups, which were the asthmatic group (Group X), the normal group (Group Z), the No. 1 sequential therapy group (Group A1), the No. 2 sequential therapy group (Group A2), the No. 3 sequential therapy group (Group A3), the montelukast group (Group M), and the budesonide group (Group B). The asthmatic model was established in each group except Group Z, by sensitization with both ovalbumin (OVA) and aluminium hydroxide via injection at the 1st, 8th and 15th day in a 22-day duration, followed by OVA aerosol inhalation every other day for 8 weeks for asthma activation. At the 8th day after the asthmatic model was established, Group A1 was orally given Ma-Xing Er-Chen Tang (MXECT) during acute phase while given normal saline (NS) during catabasis and stable phase; Group A2 was given MXECT during acute phase, and given modified Jin-Shui Liu-Jun Jian (JSLJJ) during catabasis as well as given NS during stable phase; Group A3 was given MXECT during acute phase, and given modified JSLJJ during catabasis as well as given Liu-Wei Di-Huang (LWDH) Powders during stable phase;Group M was given salbutamol via aerosol inhalation after stimulation, while orally given montelukast during catabasis and stable phase; Group B was given salbutamol via aerosol inhalation after stimulation, while given inhaled budesonide during catabasis and stable phase; Group X was given NS. After the 7-week intervention, the immunohistochemistry, western blot and real-time quantitative PCR were applied to analyze the location and quantitative expression of MMP-9 and TIMP-1, so as to find out the biological mechanism on expressions of MMP-9 and TIMP-1 in lung tissues of asthmatic rats from molecular levels to gene levels. The results of immunohistochemistry, western blot and real-time fluorescent quantitative PCR showed as follows. Compared with Group Z, the contents of MMP-9 and TIMP-1 increased significantly within lung tissues in Group X. Compared with Group X, the contents of MMP-9 and TIMP-1 decreased within lung tissues of asthmatic rats in each treatment group. It was concluded that the expression of MMP-9 and TIMP-1 elevated during asthma. TCM staged sequential therapy can regulate the ratio between MMP-9 and its inhibitor so as to block the airway remodeling, by decreasing the expression of MMP-9 and its inhibitor within lung tissues in asthmatic rats. This is one of the important action mechanisms.

Objective To investigate the effects of miR?23a?3p on proliferation, migration and apoptosis on human acute myeloid leukemia ( AML) cells by targeting SMC1A. Methods Microarray analysis was used to screen differentially expressed microRNAs and mRNAs in human AML cells. Real?time fluorescence quantitative PCR (RT?qRCR) was used to detect the expressions of miR?23a?3p and SMCA in human AML cell line U937. TargetScan database was used to analyze the correlation between miR?23a?3p and SMC1A. Double luciferase reporter gene was used to detect the interaction between miR?23a?3p and SMC1A. The effect of miR?23a?3p expression on the proliferation of U937 cells was detected by clonal assay. The migration, apoptosis, cell cycle and caspase?3 activity of U937 cells regulated by miR?23a?3p were detected by cell scratch assay and flow cytometry, respectively. Western blot was used to detect the expressions of Bax and Bcl?2 in U937 cells. Results Compared with human normal monocyte SC group (1.00), the expression of miR?23a?3p in U937 cells was up?regulated (2.56±0.78) ( P＜0.01), while the expression of SMC1A was down?regulated (0.48±0.56, P＜0.01). miR?23a?3p specifically bond to SMC1A 3′UTR and regulated the expression activity of SMC1A. Overexpression of miR?23a?3p promoted the proliferation and migration of U937 cells and inhibited the apoptosis of U937 cells, while up?regulation of SMC1A inhibited the proliferation and migration of U937 cells and promoted the apoptosis of U937 cells. The percentages of G0／G1 phase, G2／M phase and S phase cells in the negative control group were ( 37.48 ± 0.21)%, (16.78±0.18)% and (45.74±0.15)% respectively, and those in the miR?23a?3p mimics group were (19.96±0.11)%, (41.69±0.24)% and (38.24±0.34)%, respectively. The difference was statistically significant (all P＜0.05). The proportions of G0／G1 phase, G2／M phase and S phase cells in the group of miR?23a?3p mimics+pcDNA3.1?SMC1A were (36.88± 0.21)%, ( 30.44± 0.33)% and ( 32.88± 0.16)%, respectively, without significant difference when compared with those of the miR?23a?3p mimics group ( P＞0.05). The relative expression levels of Bax and Bcl?2 protein in the negative control group were 0.55±0.45 and 0.31±0.54, respectively. Overexpression of miR?23a?3p inhibited the expression of Bax protein in U937 cells (0.23± 0.13, P＜0.001), promoted the expression of Bcl?2 protein ( 0.50 ± 0.23, P＜0.01), while SMC1A increased the expression of Bax protein in U937 cells (0.40± 0.11, P＜0.01), and inhibited the expression of Bcl?2 protein (0.37± 0.15). In the negative control group, caspase?3 activity was (25.82± 0.89)%.Overexpression of miR?23a?3p inhibited caspase?3 activity in U937 cells (3.64±0.56)%, P＜0.01, while up?regulation of SMC1A promoted caspase?3 activity in U937 cells ( 15.29 ± 0.85)%, P＜0.01. Conclusion miR?23a?3p can inhibit the proliferation and migration and promote apoptosis of human AML cells by targeting SMC1A.

Objective To investigate the effects of miR?23a?3p on proliferation, migration and apoptosis on human acute myeloid leukemia ( AML) cells by targeting SMC1A. Methods Microarray analysis was used to screen differentially expressed microRNAs and mRNAs in human AML cells. Real?time fluorescence quantitative PCR (RT?qRCR) was used to detect the expressions of miR?23a?3p and SMCA in human AML cell line U937. TargetScan database was used to analyze the correlation between miR?23a?3p and SMC1A. Double luciferase reporter gene was used to detect the interaction between miR?23a?3p and SMC1A. The effect of miR?23a?3p expression on the proliferation of U937 cells was detected by clonal assay. The migration, apoptosis, cell cycle and caspase?3 activity of U937 cells regulated by miR?23a?3p were detected by cell scratch assay and flow cytometry, respectively. Western blot was used to detect the expressions of Bax and Bcl?2 in U937 cells. Results Compared with human normal monocyte SC group (1.00), the expression of miR?23a?3p in U937 cells was up?regulated (2.56±0.78) ( P＜0.01), while the expression of SMC1A was down?regulated (0.48±0.56, P＜0.01). miR?23a?3p specifically bond to SMC1A 3′UTR and regulated the expression activity of SMC1A. Overexpression of miR?23a?3p promoted the proliferation and migration of U937 cells and inhibited the apoptosis of U937 cells, while up?regulation of SMC1A inhibited the proliferation and migration of U937 cells and promoted the apoptosis of U937 cells. The percentages of G0／G1 phase, G2／M phase and S phase cells in the negative control group were ( 37.48 ± 0.21)%, (16.78±0.18)% and (45.74±0.15)% respectively, and those in the miR?23a?3p mimics group were (19.96±0.11)%, (41.69±0.24)% and (38.24±0.34)%, respectively. The difference was statistically significant (all P＜0.05). The proportions of G0／G1 phase, G2／M phase and S phase cells in the group of miR?23a?3p mimics+pcDNA3.1?SMC1A were (36.88± 0.21)%, ( 30.44± 0.33)% and ( 32.88± 0.16)%, respectively, without significant difference when compared with those of the miR?23a?3p mimics group ( P＞0.05). The relative expression levels of Bax and Bcl?2 protein in the negative control group were 0.55±0.45 and 0.31±0.54, respectively. Overexpression of miR?23a?3p inhibited the expression of Bax protein in U937 cells (0.23± 0.13, P＜0.001), promoted the expression of Bcl?2 protein ( 0.50 ± 0.23, P＜0.01), while SMC1A increased the expression of Bax protein in U937 cells (0.40± 0.11, P＜0.01), and inhibited the expression of Bcl?2 protein (0.37± 0.15). In the negative control group, caspase?3 activity was (25.82± 0.89)%.Overexpression of miR?23a?3p inhibited caspase?3 activity in U937 cells (3.64±0.56)%, P＜0.01, while up?regulation of SMC1A promoted caspase?3 activity in U937 cells ( 15.29 ± 0.85)%, P＜0.01. Conclusion miR?23a?3p can inhibit the proliferation and migration and promote apoptosis of human AML cells by targeting SMC1A.

With good prognosis of differentiated thyroid cancer (DTC), the 10-year survival rate of DTC patients is more than 90%. As a kind of radiation exposure, radioactive iodine (RAI) treatment has the potential to induce malignancies. Based on this view, whether RAI treatment will increase the risk of secondary primary malignancy (SPM) still has a lot of controversy. This review summarizes current situation of related researches, and also summarizes the limitations of the current researches and the problems to be solved in the future research. In this review, it is believed that RAI treatment does not increase the overall risk of SPM in postoperative-DTC patients.

OBJECTIVE1-Bromo-3-chloro-5,5-dimethylhydantoin (BCDMH) is a solid oxidizing biocide for water disinfection. The objective of this study was to investigate the toxic effect of BCDMH on zebrafish.

METHODSThe developmental toxicity of BCDMH on zebrafish embryos and the dose-effect relationship was determined. The effect of BCDMH exposure on histopathology and tissue antioxidant activity of adult zebrafish were observed over time.

RESULTSExposure to 4 mg/L BCDMH post-fertilization was sufficient to induce a number of developmental malformations, such as edema, axial malformations, and reductions in heart rate and hatching rate. The no observable effects concentration of BCDMH on zebrafish embryo was 0.5 mg/L. After 96 h exposure, the 50% lethal concentration (95% confidence interval (CI)) of BCDMH on zebrafish embryo was 8.10 mg/L (6.15-11.16 mg/L). The 50% inhibitory concentration (95% CI) of BCDMH on hatching rate was 7.37 mg/L (6.33-8.35 mg/L). Histopathology showed two types of responses induced by BCDMH, defensive and compensatory. The extreme responses were marked hyperplasia of the gill epithelium with lamellar fusion and epidermal peeling. The histopathologic changes in the gills after 10 days exposure were accompanied by significantly higher catalase activity and lipid peroxidation.

CONCLUSIONThese results have important implications for studies on the toxicity and use of BCDMH and its analogs.

Animals ; Antioxidants ; metabolism ; Disinfectants ; toxicity ; Dose-Response Relationship, Drug ; Embryo, Nonmammalian ; drug effects ; Hydantoins ; toxicity ; Time Factors ; Water ; chemistry ; Water Pollutants, Chemical ; toxicity ; Zebrafish

OBJECTIVETo screen a new strain which can transform panaxadiol saponins into the rare ginsenoside compound K.

METHODThe total saponins in stems and leaves of Panax notoginseng was used as a substrate in the liquid state fermentation process, and the results were detected by TLC and HPLC-ELSD to screen a strain from twelve plant pathogenic fungi which can produce ginsenoside compound K.

RESULTFusarium moniliforme was found to transform the total saponins to ginsenoside compound K efficiently in the all twelve fungal strains. In the fermentation process, ginsenoside Rb1 was transformed almost completely, and the content of ginsenoside Rd was decreasing evidently.

CONCLUSIONF. moniliforme is selected as a new high-yield strain. It is expected to be used to produce the high activity infrequent ginsenoside compound K and to improve the content of active principles in medicinal plants.

Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Fungi ; pathogenicity ; Ginsenosides ; chemistry ; Panax notoginseng ; chemistry ; microbiology