OBJECTIVE: To investigate the protective effect of selenium on toxicity of rat kidney injury caused by chronic cadmium exposure and its possible mechanism. MATERIALS: The forty-eight male SD rats were randomly divided into four groups:blank control,cadmium exposure,negative control,and selenium treatment groups. There were 12 rats in each group,and animals were given deionized water,cadmium chloride,sodium selenite,and co-processing with cadmium chloride and sodium selenite by gastric gavage,respectively,for six days a week for twelve weeks. After treatment,blood,urine and kidney tissues were collected to determinate renal function indexes,such as urea nitrogen,creatinine,and total protein in serum and urine. We observed the pathological changes and detected level and/or activity of ROS (reactive oxygen species),MDA (malondialdehyde),T-SOD (total superoxide dismutase),and NAG (N-acetyl-beta-d-glucosidase) in renal tissue. The protein imprinting method was used to detect protein expression of SETD6,DJ-1,and Nrf2. RESULTS: Biochemical and pathological results show that cadmium induced renal injury in rats,caused glomerular swelling,generalized lesions of renal tubules,and renal interstitial congestion,inflammatory cell infiltration. The levels of ROS and MDA in the kidney tissues increased (P < 0.05),and activities of T-SOD decreased (P < 0.05),indicating that cadmium exposure induced oxidative stress injury in the renal tissue. However,after selenium intervention,the renal tissue structure and functional damage caused by cadmium exposure were reduced,indicating that selenium had protective effect on kidney injury caused by cadmium exposure. At the same time,compared with the blank control,SETD6,DJ-1 and Nrf2 showed varying degrees of reduced protein expression after cadmium exposure (P < 0.05). After selenium intervention,compared to cadmium exposure group,SETD6 protein expression was significantly down-regulated (P < 0.05),DJ-1 and Nrf2 protein expression were significantly up-regulated (P < 0.05),kidney damage was reduced,suggesting that selenium might have played a protective role by inhibiting the expression of SETD6 and enhancing the expression of DJ-1 and Nrf2. CONCLUSION: Selenium effectively reduced the rat renal toxicity induced by chronic cadmium exposure. Its mechanism might be that under oxidative stress condition,selenium promoted the expression of DJ-1 by inhibiting SETD6,and thus upregulated the expression of antioxidant transcription factor Nrf2,through which enhanced the antioxidant capacity,and reduced the oxidative stress injury which was induced by cadmium exposure.

OBJECTIVE: Study the effects of nickel nanoparticles (Ni NPs) on apoptosis in SD rat Sertoli-germ cell culture and research on related mechanisms. METHODS: We established a rat Sertoli-germ cell co-culture system in vitro and the cells were treated with Ni NPs. Survive rates were determined by using the MTT assay. Apoptosis morphology were determined by Hoechst33258. Apoptosis index of Sertoli cells and germ cells were determined by Annexin V/PI staining;lncRNA expression profiles were detected to screen out the apoptosis-related genes. RESULTS: The survival rates in the Ni NPs exposure groups (25,50,100,200,400 and 800 μg/mL) were significantly lower than that in the control group (0 μg/mL) after 24 h (P < 0.01). The survival rates decreased as the concentrations of Ni NPs increased. As the Hochest33258 shows,the treatment groups showed the dense granular luminescence and obvious nuclear morphological changes in the nucleus or cytoplasm. The apoptosis indexes were 8.80%±0.50%,11.00%±0.70%,12.53%±0.71%,15.95%±0.54% and 17.80%±0.76% as the doses were 25,50,100,200,400 μg/mL. Apoptosis indexes increased as the concentrations of Ni NPs increased. The apoptosis indexes in the Ni NPs exposure groups were significantly higher than that in the control group (P < 0.01). Compared with the control group,there were 169 lncRNA (117 up and 52 down) associated with apoptosis in the exposure group of 100 μg/mL. CONCLUSION: The study shows that Ni NPs can cause apoptosis of SD rat Sertoli-germ cell co-culture with significant cytotoxicity. LncRNA were closely related to the apoptosis of SD rat co-culture Sertoli-germ cell.

OBJECTIVE: Construction of a recombinant lentiviral vector overexpressed SREBP-1c,its transfection into human hepatocellular carcinoma cell lines (HepG2) with high transfection efficiency and its effect on cellular lipid storage. METHODS: SREBP-1c gene was cloned from pCMV5-HA-SREBP-1c by polymerase chain reaction and identified by gene sequencing. The SREBP-1c gene was cloned to lentiviral expression vector (pLenti6.3/V5) by recombing DNA technology. The 293T cells were co-transfected with lentiviral packaged systems and the SREBP-1c plasmid by Lipofectamine2000 reageant to package the lentiviral particles,viral supernatant was collected,HepG2 cells were then infected by pLenti6.3-SREBP-1c. After puromycin screening,HepG2 cells were constructed. Real time PCR and Western blot assays were performed to analyzed SREBP-1c expression levels. Cellular lipid storage was detected under fluorescent microscopy and the triacylglyceride(TAG) content by TLC analysis in SREBP-1c-overexpressing HepG2 cells. RESULTS: pLenti6.3-SREBP-1c was successfully constructed and verified by restriction analysis and sequencing. SREBP-1c gene expression level in HepG2 cells with SREBP-1c gene transfection was 11.4 times higher than the control. SREBP-1c protein level in HepG2 cells with SREBP-1c gene transfection was 3.2 times higher than the control. The number and size of lipid droplets (LDs) was increased,and a higher concentration of TAG was found in SREBP-1c-overexpressing cells compared with the control (P < 0.01). CONCLUSION: Recombinant lentiviral vector pLenti6.3-SREBP-1c and SREBP-1c-overexpressing cell strain was successfully constructed. The high protein and mRNA levels of SREBP-1c were assessed after HepG2 cells were transfected. SREBP-1c increased the lipid storage of HepG2 cells. The constructed cells can become a useful tool for lipid metabolism research of liver carcinoma.

OBJECTIVE: To investigate the protective effect of allicin on cisplatin (DDP)-induced oxidative damage in HEK293 cells. METHODS: HEK293 cells were cultured in vitro and organized into different experimental groups:control,DDP treatment,allicin treatment,DDP+allicin treatment. Cell survival and expression of cytotoxicity were determined by using the MTT method. Superoxide dismutase (SOD) activity was examined by the xanthine oxidase method,malondialdehyde (MDA) content by the thiobarbituric acid method and glutathione (GSH) content by the nitrobenzoic acid method. RESULTS: Cell survival rate decreased gradually when treated with DDP alone,and this effect was dose-dependent. After pretreatment with catechin and then with DDP (20 mg/L),the cell survival rate improved significantly (P < 0.01). Allicin treatment significantly increased the intracellular MDA content,but decreased the SOD activity and GSH level compared with that in the DDP control group (P < 0.01). CONCLUSION: Allicin can antagonize DDP-induced oxidative damage in HEK293 cells and has obvious protective effects on DDP-induced toxicity in kidney cells.

OBJECTIVE: We tested the ability of five candidate compounds-isoflavones,resveratrol,quercetin,curcumin,and green tea polyphenols-to protect SH-SY5Y cells from atrazine (ATR) via in vitro experiments. METHODS: The cells were incubated for 24 h in the presence of various doses (5,10,20,50,and 100 μmol/L) of isoflavones,resveratrol,quercetin,curcumin,or green tea polyphenols. Cells were then cultured for 24 h with 250 μmol/L ATR. At the indicated time after treatment,the CCK-8 method was used to test for cell viability. Apoptosis was detected by using Annexin V-FITC and PI double dye,oxygen radical level was detected by using DCFH-DA method,and the protein expression of tyrosine hydroxylase (TH) by using Western blot. RESULTS: Five plant compounds showed significant anti-ATR effect at low doses,isoflavones 5 μmol/L,resveratrol 5 μmol/L,quercetin 50 μmol/L,curcumin 5 μmol/L and green tea polyphenols 20 μmol/L. Compared with other four plant compounds,the effect of isoflavones anti-ATR injury on SH-SY5Y was significant,especially in 5 μmol/L (P < 0.05). And isoflavones (5 μmol/L) could significantly reduce the level of ROS and increase the expression of TH (P < 0.05). CONCLUSION: Isoflavones could protect SH-SY5Y cells from ATR by reducing intracellular ROS levels,increasing the expression of TH and cell viability to inhibit cell apoptosis.

OBJECTIVE: To observe the effects of chlorine on structure and function of mitochondria of lungs in rats and explore the protective effects of salidroside on acute lung injury induced by chlorine. METHODS: 36 healthy male Sprague-Dawley (SD) rats were divided into 6 groups:1 control and 5 exposure. Rats in the exposure groups were exposed to an initial concentration of 1 200 mg/m³ chlorine for 5 min in a devised cabinet or room air. Rats were sacrificed at 0.5,1.5,3,6 and 9 h,respectively,after exposure. In addition,24 rats were randomly divided into 4 groups:control,chlorine,chlorine+salidroside and salidroside. Chlorine and chlorine+salidroside groups were exposed to chlorine as mentioned above. Chlorine+salidroside and salidroside groups were treated with salidroside (300 mg/kg) at 30 min before chlorine exposure and at 15 min after chlorine exposure. Then the rats were sacrificed at 3 h after chlorine exposure,and the lung tissues were harvested. Lung mitochondria ultra structures were observed with electron microscope. The Na⁺,K⁺-ATPase,Ca²⁺,Mg²⁺-ATPase in lung tissues and lactate dehydrogenase (LDH) in serum and bronchoalveolar lavage fluid (BALF) were measured using kits,respectively. The mitochondria in lung tissues were isolated with differential centrifugation. The protein levels of Tom 20,PINK1 and Parkin in lung tissues or mitochondria were detected by western blotting. RESULTS: Compared with control group,LDH of BALF in exposure groups were significantly increased (P < 0.05). Tom 20,PINK1 and Parkin protein expressions in lung tissues were all higher than that in control group (P < 0.05). Expression of Tom 20 was highest at 3 h. It was also shown by transmission electron microscope that the mitochondria structure in the control group was normal. The mitochondria in rat pulmonary vascular endothelial cells were swollen with disrupted or disintergrated cristae. When treated with salidroside,the expression levels of PINK1 and Parkin in lung tissues and PINK1 in mitochondria were reduced compared with chlorine group at 3 h. The expressions of Parkin in mitochondria was markedly increased (P < 0.05);Na⁺,K⁺-ATPase,Ca²⁺,Mg²⁺-ATPase in lung tissues and LDH in serum were decreased. CONCLUSION: Chlorine induced mitochondrial structural damage and functional impairment in rats and salidroside exerted a beneficial effect through protecting the mitochondria.

OBJECTIVE: To investigate the effects of isoimperaterin on regulation of tyrosinase and Rab27a in cultured human epidermal melanocytes. METHODS: Human epidermal melanocytes were isolated and cultured. The cells were randomly divided into control and experimental groups. Melanocytes of the control group were cultured with M-254 culture medium. Cells of the experimental group were cultured with M-254 culture medium containing 25 μmol/L isoimperaterin. The cells were cultured for 48 h and observed using inverted microscopy. Cell vitality was assayed using MTT method. The expression of tyrosinase and transfer-related protein Rab27a were analyzed by western blotting. The activity of tyrosinase and the content of melanin were measured by L-dopa oxidation and NaOH method,respectively. RESULTS: Compared with the control group,the dendrites of melanocytes of experimental group were elongated after treatment with 25 μmol/L isoimperaterin for 48 h. Melanocyte vitality was reduced by 17.2% (P < 0.05). Expression of tyrosinase was decreased by 30.3% (P < 0.01) and Rab27a was increased by 31.9% (P < 0.05). The activity of tyrosinase was reduced by 25.6% (P < 0.05). Melanin content was decreased by 30.9% (P < 0.01). CONCLUSION: 25 μmol/L isoimperaterin elongated melanocytic dendrites but inhibited melanocyte vitality. It also inhibited the expression and activity of tyrosinase and promoted the expression of transfer-related protein Rab27a to reduce the content of melanin in cultured human epidermal melanocytes.

OBJECTIVE: To investigate expression of apoptosis-related genes during different dose of X-ray-induced bystander effects in rats. METHODS: 70 male Wistar rats were randomly divided into the normal control group and the different irradiation groups. For the latter groups,the right lungs of the rats were radiated with 2,4,8 Gy X-rays. During irradiation,the rest of the body was covered with lead shielding. At 6,24 and 48 h respectively after radiation,7 rats of each radiation group were executed. Variations in mRNA expression of Cytochrome C,Caspase-9,Caspase-3,Bax and Bcl-2 in the right lung,left lung and left kidney tissue were detected with quantitative polymerase chain reaction (qPCR). RESULTS: The mRNA expressions of Cytochrome C,Caspase-9 and Caspase-3 in the right lung,left lung and left kidney tissue of the irradiated groups were up-regulated at all time points. However,the mRNA expressions of Bcl-2 were down-regulated at all time points in the right and left lungs. The observed changes were significantly different in comparison with the normal control group (P < 0.05 or P < 0.01). In the right lung,expressions of Bax mRNA were significantly increased in all irradiated groups except the 8 Gy dose group at 48 h (P < 0.05 or P < 0.01). In the left lung,expressions of Bax mRNA were significantly increased in each radiation group (P < 0.01). In the left kidney,expressions of Bax mRNA were significantly increased in all irradiated groups except the 2 Gy dose group at 6 h. Compared with the normal control group,the differences were statistically significant (P < 0.01). Expressions of Bcl-2 mRNA were decreased at all time points after irradiation except the 8 Gy dose group at 6 h and the difference was statistically significant compared with the normal control group (P < 0.05 or P < 0.01). CONCLUSION: After the right lungs of the rats were irradiated by different doses of X-rays,expressions of Cytochrome C,Caspase-9,Caspase-3 and Bax in the right lung tissue were up-regulated. While expressions of Bcl-2 were down-regulated. The left lung and left kidney tissues that were not irradiated also responded to the X-rays. And the radiation-induced bystander effects were observed in vivo. Between every radiation dose and every time point,there were no apparently different effects.

OBJECTIVE: To study the effects of expression of HIF-1α on cytotoxicity of K562 cells induced by 1,4-benzoquinone (1,4-BQ),hydroquinone (HQ) and phenol. METHODS: The control K562 cells (K-c) and K562 cells with high expression of HIF-1α (K-h) were treated with different concentrations of 1,4-BQ (0,10,20,40,80 μmol/L),HQ (0,10,20,40,80 μmol/L) and phenol (0,1,1.5,2,2.5,5 mmol/L) for 24 h. Cell proliferation was detected by MTT assay,apoptosis was detected by flow cytometry with PI/FITC Annxein V double staining,and cell cycle was detected by propidium iodide combined with flow cytometry. RESULTS: The MTT results revealed that the relative proliferation rate of K-c cells and K-h cells showed a dose-related decrease after treating with 1,4-BQ (P < 0.05). The relative proliferation rate of K-h was higher than control K-c (P < 0.05) in 80 μmol/L 1,4-BQ. After treating with HQ and phenol at higher concentrations,K-c exhibited proliferation inhibition,while no significant proliferation inhibition was observed in K-h. Flow cytometry results revealed that the apoptosis rate increased in K-c and K-h (P < 0.05) and showed a dose-response relationship after exposuring to 1,4-BQ for 24 h (P < 0.05). Moreover,the apoptosis rate of K-hwas lower than K-c (P < 0.05) with 20 μmol/L 1,4-BQ. After exposure to HQ and phenol,K-c and K-h demonstrated no significant differences compared with control group,K-h was not significantly different from K-c. The results of cell cycle analysis demonstrated K-c were arrested in G0/G1 phase or S-phase after exposure to 1,4-BQ,HQ and phenol. CONCLUSION: Among benzene metabolites,1,4-BQ was the most cytotoxic,followed by HQ and phenol. High expression of HIF-1α reduced the cytotoxicity of K562 cell caused by benzene metabolites,and HIF-1α might work in regulating cell proliferation,apoptosis through cell cycle control.

OBJECTIVE: To compare the sub-chronic toxicity caused by three scales of nano-zinc oxide and micron-zinc oxide in rats. METHODS: SPF SD rats were randomly divided into 5 groups with 8 rats (four females and four males) per group. Once a day for 42 days,rats received intraperitoneal injection of the same dose (10 mg/kg) of different scales nano-ZnO(30 nm,50 nm,100 nm) and micron-ZnO(≤ 1 μm). The negative control group was injected with saline. At the end of the experiment, organ coefficients of the liver,spleen,lung,kidney and testis;haematology and blood biochemical parameters;the sperm abnormality rate of the epididymis were determined. The main organs were used to prepare for pathological sections and HE staining. TUNEL method was used to detect testicular germ cell apoptosis. RESULTS: Compared with the negative control group,there were different degree of changes to the organ coefficients of liver,spleen,kidney and lung in the three scale nano-and micron-ZnO groups. Compared with the micron-ZnO group,the organ coefficients of spleen in 30 nm and 50 nm ZnO groups were all increased,while the organ coefficients of lung in 30,50 and 100 nm groups were decreased (P < 0.05). The number of erythrocytes (RBC),neutrophil ratio (NEUT,%) and platelet count (PLT) in the three scale of nano and micron-ZnO groups were significantly higher than the negative control group,while the lymphocyte ratio was significantly decreased (P < 0.05). The number of PLT in the 30 and 50 nm ZnO groups was significantly higher than that in the micron-ZnO group (P < 0.05),but the hemoglobin (HGB) was decreased (P < 0.05). The sperm abnormality rate induced by 30nm and 50nm ZnO group was 1.05% and 0.81%,respectively,and it was statistically significant (P < 0.05) compared with the negative control group(0.24%). The number of spermatogenic cells on the wall of the testis tissue of the three scale of nano-ZnO group was significantly lower than that of the control group. Apoptosis of the spermatogenic cells was observed by TUNEL method,and the apoptosis frequency of the nano-ZnO group was statistically different (P < 0.05). CONCLUSION: The nano-ZnO caused higher sub-chronic toxicity than micron-ZnO. The target organs of nano-ZnO included the testicle,liver,bone marrow and kidney. With the decrease of particle sizes,the number of affected organs and the degree of damage increased.

OBJECTIVE: This paper was to study on the effects of rhizosphere growth promoting bacteria Paenibacillus polymyxa on strontium enrichment in rape. From the aspect of food safety assessment,it was of practical significance to explore the characteristics of enrichment and transfer of typical radionuclide strontium in vegetables. It also provided a theoretical basis for the feasibility of phytoremediation of contaminated soil. METHODS: The experiment was divided into 5 groups:4 different strontium concentration treatment groups and 1 control groups (hydroponics blank control). The vegetable was cultivated by hydroponic method and Paenibacillus polymyxa was added. Vegetables were harvested at 3 time points (7 d,14 d,21 d) after adding Paenibacillus polymyxa to the hydroponic solution. Then the concentration of strontium in roots,stems and leaves of rape was determined by inductively coupled plasma atomic emission spectrometry (ICP-AES) and the concentration ratios and transportation factors were calculated. RESULTS: Compared with blank control groups,when Paenibacillus polymyxa was present,strontium concentrations in the stems,leaves and roots of rape increased with strontium concentration in hydroponic solution and increased with the number of cultivation days (P < 0.05) (except rape roots of 1 mmol/L strontium concentration in hydroponic solution with 21 d). The content of strontium in stems and leaves of rape was greater than that in rape roots (P < 0.05). Paenibacillus polymyxa led to reduction of the concentration ratios of strontium but to increase of the transportation factors which promoted the transfer of strontium from root to stem and leaf. TF(transportation factor) of strontium increased in experiment groups 1,2,3 and 4,with increased number of days to reach the highest in 21 days. CONCLUSION: Paenibacillus polymyxa significantly affected the transfer of strontium in rape. It reduced the concentration ratios of strontium in roots,stems and leaves of rape,and promoted the strontium transfer from root to stem and leaf.

OBJECTIVE: To investigate the effectiveness of transcarotid artery chemotherapy on gliomas in rats. METHODS: After the brain glioma model was established successfully,rats were randomly divided into control group(the same amount of saline injected through the tail vein),intravenous injection of cisplatin group (V+CDDP),intravenous bevacizumab group (V+BVZ),arterial injection of cisplatin group (A+CDDP),arterial injection of bevacizumab group (A+BVZ). After treatment for 7 d,the glioma volume was detected,and the expression of VEGF,P21,Bcl-2 and MMP-2 protein and mRNA were detected by Western blot and real time-PCR. RESULTS: The glioma volume in A+CDDP group[(5.58±0.37) mm³] was smaller than that in V+CDDP group[(7.67±0.70) mm³]. The glioma volume in A+BVZ group[(4.81±0.20) mm³] was smaller than that in V+BVZ group[(6.57±0.69) mm³]. Differences were statistically significant (P < 0.01),and it was shown that artery injection had a strong inhibitory effect on the size of glioma. At the same time,the glioma volume in V+BVZ group was smaller than that in V+CDDP group (P < 0.01),the glioma volume in A+BVZ group was smaller than that in A+CDDP group (P < 0.05). BVZ had a stronger inhibitory effect on glioma size compared with CDDP,and artery injection had a stronger inhibitory effect than intravenous injection on the size of glioma. Compared with intravenous injection,the expression of VEGF,Bcl-2 and MMP-2 protein and mRNA were significantly decreased (P < 0.05) and the expression of P21 protein and mRNA was significantly increased in arterial injection group (P < 0.05). Compared with CDDP,the expression of VEGF,Bcl-2 and MMP-2 protein and mRNA were significantly decreased (P < 0.05) and the expression of P21 protein and mRNA was significantly increased (P < 0.05) in BVZ group. CONCLUSION: Application of BVZ transcatheter arterial chemoinfusion effectively inhibited the size of glioma,therefore,this approach may be useful for providing better therapeutic effect.

OBJECTIVE: This paper aims to study the mutation of epidermal growth factor receptor (EGFR) gene in fresh cytological specimens from patients with lung adenocarcinoma,and to determine the prognosis of positive patients by tyrosine kinase inhibitor (TKI). METHODS: A total of 313 specimens from needle aspiration and pleural effusion were collected in the Cancer Detection Center of the Fourth Hospital of Hebei Medical University. After HE and immunocytochemistry stainings,the specimens were diagnosed as lung adenocarcinoma by two cytology pathologists. The mutation of 18-21 exon was detectied using ARMS to observe mutations situation. Then, the objective response rate (ORR) and the progression-free survival (PFS) between the targeted group and the chemotherapy group of patients were comparedith. RESULTS: Among 313 cases,293 cases of lung adenocarcinoma were diagnosed,and DNA specimens were extracted from 288 cases,the success rate was about 98.3%. 130 mutations were found and the rate was 45.1%. EGFR mutation of adenocarcinoma patients mainly occurred to females,nonsmokers,but had nothing to do with age. The ORR was statistically different between the targeted group with chemotherapy (P < 0.01),and PFS curve of targeted group was on chemotherapy group. The efficacy and the survival time of targeted group and targeted and chemotherapy group were superior to that of chemotherapy group. The results of the EGFR mutation and the prognosis of the tested positive patients in the fresh cytology samples were consistent with that from previous literatures. CONCLUSION: The results of the test were accurate,and fresh cytological specimens can be used as a replacement for tumor tissue specimens.

OBJECTIVE: Genotoxicity of triclosan was evaluated in TK6 and bacterial cells by alkaline comet assay,cytokinesis-block micronucleus (CBMN) test and bacterial reverse mutation test (Ames test). METHODS: Comet assay and CBMN test were performed by TK6 cell,and 5 doses of TCS were set as 3.5,8.8,17.5,26.3 and 35 μmol/L. Three Salmonella typhimurium tester strains TA98,TA100 and YG7108 (DNA repair deficient strain,Ogt^-/Ada^-) were employed to perform Ames test,and 8 doses of TCS were set at 0.000 5,0.001 67,0.005,0.016 7,0.05,0.167,0.5 and 1.67 μg/plate. RESULTS: The results of comet assay showed that TCS caused DNA damage with a significant increase in comets tail length,tail DNA intensity and tail moment (P < 0.01),and have a statistical correlation with tail DNA intensity (r=0.943,P=0.017). The CBMN test indicated that TCS did not induce increased frequency of micronucleus (P > 0.05) and no consistent increase in numbers of revertant colonies of three strains was seen in Ames test (P > 0.05). CONCLUSION: TCS mainly induced severe DNA injury to TK6 cell in a dose-dependent manner. TCS did not cause chromosome damage and also was not mutagenic to three Salmonella typhimurium strains.