Effects of arsenite and MPST over-expressin on GRP78/CHOP endoplasmic reticulum stress pathway in SH-SY5Y cells / 中国药理学通报

Aim To investigate whether endoplasmic reticulum stress is involved in the neurotoxicity of sodi¬um arsenite and clarify whether over-expression of 3-mercaptopyruvate sulfurtransferase (MPST) regulates endoplasmic reticulum stress induced by arsenic. Methods The SH-SY5Y cell line stably expressing the exogenous MPST gene was obtained by constructing the lentiviral vector of MPST gene. The SH-SY5Y cells were randomly divided into six groups, the SR-MPST over-expression group stably expressing the exogenous MPST gene, SH-PEB control group transfected with empty vector, the arsenite treatment group ( NaAs02 group ), TUDC A treatment group ( blocker of endoplasmic reticulum stress ) and TUDC A + NaAs02 group. Western blot was used to examine the protein expression of GRP78 and CHOP after different treatment. Results Although MPST overexpression had no significant effects on the expression of GRP78 and CHOP proteins, NaAs02 could significantly increased the protein levels of GRP78 and CHOP ( P < 0. 01 ) and the up-regulation of GRP78 and CHOP proteins caused by NaAs02 could be blocked by the treatment of TUDC A. In addition, the inhibition by MPST overexpression on the arsenic-induced increase of GRP78 and CHOP proteins (P <0. 01 ) could also be reversed by the TUDC A treatment significantly. Conclusions The GRP78/ CHOP endoplasmic reticulum stress pathway is involved in the neurotoxic damage induced by arsenic; MPST overexpression may decrease arsenic-induced endoplasmic reticulum stress.

Value of texture analysis on apparent diffusion coefficient maps in the preoperative prediction of histological grade of tongue and mouth floor squamous cell carcinoma / 中华放射学杂志

Objective To explore the value of texture analysis on ADC maps in the preoperative prediction of histological grade of tongue and mouth floor squamous cell carcinoma (SCC). Methods Forty?nine pathologically confirmed tongue and mouth floor SCC with definite grading from May 2015 to June 2018 were retrospectively analyzed, including 21 cases of gradeⅠ, 21 cases of gradeⅡand 7 cases of gradeⅢ. All subjects underwent preoperative MRI examination with DWI included. Two doctors delineated whole tumor region of interest and extracted texture parameters by the 3D Slicer software, including 8 histogram parameters, 11 grey?level co?occurrence matrix (GLCM) parameters and 7 gray?level run?length matrix (GLRLM) parameters. Intraclass correlation coefficient (ICC) was used to evaluate the inter?observer delineation agreement, and the texture parameters with excellent reproducibility (ICC>0.8) were used for analysis only. Mann?Whitney U test was used to compare the differences of ADC texture parameters between grade Ⅰ and grade Ⅱ?Ⅲ SCCs. Stepwise logistic regression was used to determine the independent predictors and to build combined model. ROC analysis was used to explore the performance of texture parameter and model in predicting histological grade of tongue and mouth floor SCCs. Pearson correlation coefficient was used to evaluate the correlation between texture parameters with statistical significance. Results (1) Excellent inter?observer delineation agreement (ICC: 0.81-0.98) was observed in 69.23% (18/26) texture parameters, including 6 histogram parameters, 7 GLCM parameters and 5 GLRLM parameters. (2) Among histogram parameters, significantly higher 10 percentile ADC value (ADC10) and significantly lower energy and entropy were shown in gradeⅠcompared with gradeⅡandⅢSCCs (all P<0.05). Among GLCM parameters, significantly lower joint entropy, difference entropy, sum entropy, difference variance, difference average and contrast were shown in grade Ⅰ SCCs (all P<0.05). Among GLRLM parameters, significantly lower gray?level nonuniformity and run?length nonuniformity were shown in gradeⅠSCCs (all P<0.05). ADC10 and entropy were identified as independent predictors. The ADC10 and entropy were 960(913, 1 178)×10?6mm2/s and 4.32(4.06, 4.76) in gradeⅠSCCs, and 888(816, 987)×10?6mm2/s and 4.88(4.57, 5.29) in gradeⅡ?ⅢSCCs respectively. The area under ROC curve (AUC) of ADC10, entropy and combined model were 0.72, 0.75, 0.81. (3) Significant correlation (|r|≥0.5) was observed among 52.73% (29/55)texture parameters with statistical significance. Conclusion Texture analysis on ADC maps can provide more quantitative information, which can be more accurately in discriminating grade Ⅰfrom gradeⅡ?Ⅲtongue and mouth floor SCCs.

To compare the predictive value of the radiomics siganature extrated from MRI plain or enhancement imaging for the survival of rectal cancer / 中华放射学杂志

Objective To compare the predictive value of radiomics signature extracted from MRI plain and enhancement sequence for the disease-free survival (DFS) of rectal cancer. Methods We retrospectively analyzed fifty-one patients with rectal adenocarcinoma confirmed by biopsy from October 2010 to December 2013 in Cancer Hospital Chinese Academy of Medical Sciences.All patients underwent neoadjuvant chemotherapy(nCRT)followed total mesorectal excision(TME),and MRI scans were performed before nCRT.Follow-up time for the survival patients were more than 3 years.The image segmentation was performed on the T2WI sequence of the small FOV and the multi-phase enhancement sequence venous phase,respectively.Least absolute shrinkage and selection operator(LASSO)Cox regression was applied to extract radiomics features and the imaging signature was constructed. According to the radiomics score of each patient,the patients were divided into the high risk group with shorter DFS and the low risk group with longer DFS. A 3-year DFS was calculated for radiomics signature using the Kaplan-Meier product limit method with univariate log-rank analysis testing for differences in the training and validation cohort, respectively. And the predictive ability of the model was evaluated by concordance index (C-index). Results The training set and the validation set were 36 and 15 cases, respectively. During follow-up 32 patients experienced relapse(26 distant,3 local and 3 both),and 19 cases were censored.Twelve features were extracted in the enhanced sequence.The radiomics signatures were significant for DFS in the training set and the validation set(P=0.000 2 and 0.009 1,respectively).The C-index of the model were 0.904 and 0.700 in the training set and the validation set, respectively. The model has the better ability to predict survival.Two features were extracted in the plain sequence.The radiomic signature was significant for DFS in the training set(P=0.005 0),while the radiomics signature was not significant for DFS in the validation set (P=0.767 0). The C-index of the model were 0.711 and 0.500 in the training set and the validation set, respectively.Conclusions Radiomics signature extracted from MRI venous phase enhancement sequence superior to plain sequence for predicting the DFS of rectal cancer before nCRT.

Application and Practice of Risk Managements Analysis Methods in Medication Error Prevention in Medi-cal Institutions / 中国药师

Medication safety is one of patient safety goals, and medication error prevention is its main content. Risk managements such as hazard analysis and critical control points ( HACCP) and failure mode and effect analysis ( FMEA) with a lot of applications in health care field have a proper condition to be used for medication error prevention. A risk management was carried out for medication error prevention through the design and practice in our hospital, which contained the following 7 steps:the medication error prevention project was determined, the management team was made up, the links of medication and types of medication errors were confirmed, the critical control points were determined by using HACCP, the critical control points were analyzed by using FMEA, the links with high scores were performed improvement in prevention plan, and the improvement plan was applied and evaluated. The results showed the risk management used in medication error prevention is flexible and practical, which is especially suitable for the risk management for high-alter drugs.

Subtype discrimination of lung adenocarcinoma manifesting as ground glass nodule based on radiomics / 中华放射学杂志

Objective To develop and validate the radiomics nomogram on the discrimination of lung invasive adenocarcinoma from'non-invasive'lesion manifesting as ground glass nodule(GGN)and compare it with morphological features and quantitative imaging. Methods One hundred and sixty pathologically confirmed lung adenocarcinomas from November 2011 to December 2014 were included as primary cohort. Seventy-six lung adenocarcinomas from November 2014 to December 2015 were set as an independent validation cohort. Lasso regression analysis was used for feature selection and radiomics signature building. Radiomics score was calculated by the linear fusion of selected features. Multivariable logistic regression analysis was performed to develop models. The prediction performances were evaluated with ROC analysis and AUC,and the different prediction performance between different models and mean CT value were compared with Delong test. The generalization ability was evaluated with the leave-one-out cross-validation method. The performance of the nomogram was evaluated in terms of its calibration. The Hosmer-Lemeshow test was used to evaluate the significance between the predictive and observe values.Results Four hundred and eighty-five 3D features were extracted and reduced to 2 features as the most important discriminators to build the radiomics signatures. The individualized prediction model was developed with age, radiomics signature, spiculation and pleural indentation, which had the best discrimination performance(AUC=0.934)in comparison with other models and mean CT value(P<0.05)and showed better performance compared with the clinical model(AUC=0.743,P<0.001).The radiomics-based nomogram demonstrated good calibration in the primary and validation cohort, and showed improved differential diagnosis performance with an AUC of 0.956 in the independent validation cohort. Conclusion Individualized prediction model incorporating with age, radiomics signature, spiculation and pleural indentation, presenting with radiomics nomogram, could differentiate IAC from'non-invasive'lesion manifesting as GGN with the best performance in comparison with morphological features and quantitative imaging.

Medication Safety Analysis of Cephalosporin in Our Hospital / 中国药师

Objective:To understand the medication safety events about cephalosporin in our hospital in order to provide evidence for the better medication safety of cephalosporin. Methods:Adverse drug reaction ( ADR) reports, medication error ( ME) reports and drug quality problem records about cephalosporin in our hospital were collected and the critical indicators in the reports were analyzed. Re-sults:The ADR reports selected in our hospital were mainly reported by pharmacists. More males were involved in the related patients, and young adults were in the majority. Second and third-generation cephalosporin were the major drugs used in the reports. The damages in skin and its appendages were the major damages in organs. The ME reports selected in our hospital were totally reported by pharma-cists. Errors in frequency and dose of drug administration were the most common, and the main causes were lack of knowledge and train-ing. No serious quality problem in cephalosporin was found in our hospital, and the inner and outer packaging in damaged conditions was the most common problems in our records. Conclusion:Cephalosporin as one widely used antibacterial agent is considered safe and effec-tive. Because of its large amount of application, cephalosporin should be paid more attention by health care professionals. MEs should be avoided as far as possible. ADRs should be treated timely and properly in order to decrease medical disputes. More attention should be paid to ensure security of drug use for patients.

Analysis of the Situation of the Minimum Sales Package Unit / 中国药房

OBJECTIVE:To analyze the situation of the minimum sales package unit,and to provide reference for standardiz-ing package. METHODS:According to related regulations of New Edited Materia Medica(17th edition)and package inserts,373 kinds of oral drugs,57 kinds of oral drugs for lowering blood glucose,blood lipid and blood pressure,6 kinds of second kind psy-chotropic oral drugs were analyzed statistically in respect of minimum sales package unit and medication duration. RESULTS:There were 30 kinds of minimum sale package unit for 373 oral drugs,among which common capacity were 20,10,100 grain/tab-let/pill,accounting for 14.21%,14.21%,12.33% respectively,and 57.11% were used within 7 d. There were 13 kinds of mini-mum sale package unit for 57 oral drugs for lowering blood glucose,blood lipid and blood pressure,among which the common ca-pacity were 7,30,14,10 grain/tablet/pill,accounting for 33.34%,14.04%,14.04%,14.04% respectively,and 85.97% were used within 14 d,indicating sales package unit often based on weekly dosage. Among 6 kinds of type Ⅱ psychotropic oral drugs, minimum sales package unit capacity of 3 kinds were all 20 grain/tablet/pill,and those of other 3 kinds were 10,7,6 grain/tablet/pill;one of them were used more than 7 d. CONCLUSIONS:Minimum sales package unit is confirmed discretionarily. It is sug-gested that minimum sale package unit of oral drugs for lowering blood glucose,blood lipid and blood pressure should be packaged according to the dosage of 2 weeks to one nonth;the capacity of type Ⅱ psychotropic oral drugs is lower than the dosage of 7 days,and daily dose package is the best.

Expression of mitochondrial fission protein locus Fis1 and ultrastructural changes in the renal cells of rats with chronic fluorosis / 中国地方病学杂志

Objective To observe the expression of mitochondrial fission protein locus Fis1 and ultrastructural changes in the renal cells of rats with chronic fluorosis,and to reveal the mechanism in mitochondrial damage of the renal cells.Methods Sixty SD rats were randomly divided into 3 groups according to sex and body mass(20 in each group):control group,lower fluoride group and higher fluoride group.All the rats were fed with different doses of sodium fluoride in drinking water(0,10 and 50 mg/L,respectively).Six-month later,the expression of Fisl in renal cells was determined by real-time fluorenscence quantitative PCR and immunohistochemistry technology,the mitochondrial morphology of renal cells was observed under transmission electron microscopy (TEM).Results As compared with the control group(28.70 ± 12.41),Fis1 mRNA levels(91.48 + 34.83 and 582.09 ± 184.69) in renal cells of the lower fluoride and the higher fluoride groups were increased(all P ＜ 0.05).As compared with the control group(10.49 ± 7.66),Fisl protein levels(16.33 ± 10.26 and 21.50 ± 5.24) in renal cells of the lower fluoride and the higher fluoride groups showed a trend of increasing,the higher fluoride group was higher than that of the control group(P ＜ 0.05).By TEM,mitochondrial crest in renal cells of the lower fluoride and the higher fluoride groups was vague or disappeared,mitochondrial division section appeared.Conclusions Fluoride is a kind of toxicant that can cause damage to mitochondrion of renal cells,induce the expression of Fis1 in transcriptional and protein level,and lead to the obstacles of mitochondrial fusion-fission and ultrastructural abnormality of mitochondrion,which may play an important role in mechanism of mitochondrial damage in the renal cells of rats with chronic fluorosis.

Effects of chronic fluorosis on 4.8 kb mitochondrial DNA in liver, kidney and brain of rats / 中国地方病学杂志

Objective To investigate the deletion pattern of 4.8 kb mitochondrial DNA(mito-DNA) in liver,kidney,and brain of rats with chronic fluorosis and to explore the significance of mitochondria in the pathogenesis of chronic fluorosis.Methods Sixty SD rats were randomly divided into 3 groups according to body mass (20 in each group):control,low-fluoride and high-fluoride groups,and they were fed with different concentrations of fluoride in drinking water (0,10,50 mg/L,respectively) for 6 months.Mito-DNA in liver,kidney and brain was detected by real-time PCR.Results The amounts of 4.8 kb mito-DNA in liver(2.1 × 10-3,1.6 × 10-3),kidney (1.7 × 10-3,1.4 × 10-4) and brain cortex (1.5 × 10-5,1.3 × 10-5) in low-and high-fluoride groups were significantly reduced,as compared with that of control group (2.9 × 10-3,2.0 × 10-3,1.1 × 10-4,all P ＜ 0.05).The amount of 4.8 kb mito-DNA in kidney in high-fluoride group was lower than that in low-fluoride group (P ＜ 0.05).Conclusions Excessive fluoride intake can result in missing of 4.8 kb mito-DNA in liver,kidney and brain cortex.The abnormal of mito-DNA might be related to the dysfunction of mitochondrial respiratory chain.

Changes of syndecan-4 and nuclear factor κB in the kidney of rat with chronic fluorosis / 中国地方病学杂志

Objective Aim of the study is to investigate the expression of syndecan-4 and nuclear factor κB(NF-κB) in the kidney of rat with chronic fluorosis,and to reveal the mechanism of kidney damage resulted from the toxicity of excessive amount of fluoride.Methods According to body mass and sex,sixty SD rats were randomly divided to three groups according to body mass and fed with different contents of fluoride:control group with normal tap-water(＜ 0.5 mg/L fluoride),small dosage of fluoride exposure group (adding 10 mg/L fluoride in tap-water) and large dosage of fluoride exposure group (50 mg/L fluoride) for six months.The protein level of syndecan-4 and NF-κB in the kidney was detected by Western blotting and syndecan-4 mRNA level by quantitative real time PCR.Results As compared to the control group[(100.0 + 8.1)％],the expression of syndecan-4 at protein level in the kidney of rat was significantly increased in the small dosage of fluoride exposure group [(198.5 + 5.6)％,P ＜ 0.05] and large dosage of fluoride exposure group [(209.2 + 13.0)％,P ＜ 0.05]; the protein levels of NF-κB in the small dosage of fluoride exposure group[(284.4 + 11.1)％,P ＜ 0.05] and in the large dosage of fluoride exposure group[(343.2 + 2.9)％,P ＜ 0.05] were significantly increased than that of the control group[(100.0 ± 10.7)％].The mRNA levels of syndecan-4 in the kidney in the small dosage of fluoride exposure group and large dosage of fluoride exposure group(0.431 + 0.058 and 0.453 ± 0.065,both P ＜ 0.05,respectively) were significantly increased than that of the control(0.128 + 0.026).Conclusions The increased expression of NF-κB in the kidney is induced by increased expression of syndecan-4,which may be involved in kidney damage of chronic fluorosis.

Expression of receptor for advanced glycation endproducts and nuclear factor κB in brain hippocampus of rat with chronic fluorosis / 中国地方病学杂志

Objective To investigate the expressions of receptor for advanced glycation endproducts (RAGE) and nuclear factor κB(NF-κB) in brain hippocampus of rat with chronic fluorosis,and to reveal the mechanism of brain damage resulted from chronic fluorosis.Methods Sixty clean grade SD rats were randomly divided to three groups(20 rats in each group,10 female and 10 male) fed with different contents of fluoride,control group with normal tap-water(＜ 0.5 mg/L fluoride),small dosage of fluoride exposure group(10 mg/L fluoride in tap-water) and large dosage of fluoride exposure group(50 mg/L fluoride) for six months.Then the rats were killed by femoral artery bleeding and hippocampus was removed.Protein and mRNA levels of RAGE and NF-κB in the hippocampus were determined by Western blotting and quantitative real time PCR,respectively.Results As compared to the control groups[(100.00 ± 2.60)％,(100.00 ± 7.80)％],the expressions of RAGE and NF-κB at protein level in the hippocampus were significantly increased in the small dosage of fluoride exposure groups [(205.00 ± 15.30)％,(156.00 ± 12.20)％] and the large dosage of fluoride exposure groups[(232.00 ± 10.90)％,(162.00 ± 9.80)％,all P ＜ 0.05]; for the mRNA level of RAGE and NF-κB,the expressions were higher in the small dosage of fluoride exposure groups(1.27 ± 0.09,0.83 ± 0.15) and the large dosage of fluoride exposure groups (2.60 ± 0.19,1.27 ± 0.19) than those of the control groups(0.66 ± 0.18,0.32 ± 0.08,all P＜ 0.05).Conclusions The increased expressions of RAGE and NF-κB in the hippocampus of rat brain are caused by chronic fluorosis,and these changes may be associated with the mechanism of nerve injury.

Changed expression of mito-fusion 1 and mitochondrial fragmentation in the cortical neurons of rats with chronic fluorosis / 中华预防医学杂志

<p><b>OBJECTIVE</b>To observe the mitochondrial fragmentation and the expression of mito-fusion 1 gene in the cortical neurons of rats with chronic fluorosis, and to reveal their roles in mitochondria damage to neurons due to chronic fluorosis.</p><p><b>METHODS</b>SD rats were divided randomly into three groups of 20 each (a half females and a half males housed individually in stainless-steel cages), and fed with the different doses of fluoride containing in drinking water (untreated control containing 0 mg/L fluoride, and low-fluoride and high supplemented with 10 and 50 mg/L fluoride, respectively). After 3 or 6 months exposure, the mitochondrial morphology of the neurons in rat brains were observed by transmission electron microscopy (TEM), then the expression of mitochondrial fusion gene, Mfn1, were detected by immunohistochemistry and western-blotting, respectively.</p><p><b>RESULTS</b>Dental fluorosis was obvious in the rats exposed to excessive fluoride in their drinking water, that is, (16 rats out of 20) numbers of I° detal fluorosis in the low-fluoride group, and (11 rats out of 20) numbers of I° and (9 rats out of 20) numbers of II° detal fluorosis in the high-fluoride group were observed after 3 months exposure. Moreover, (14 rats out of 20) numbers of I° and (6 rats out of 20) numbers of II° detal fluorosis in the low-fluoride group and (6 rats out of 20) numbers of Io, (13 rats out of 20) numbers of II°, and (1 rats out of 20) numbers of III° detal fluorosis in the high-fluoride group were observed after 6 months exposure. And both of untreated controls without detal fluorosis were also observed. The urinary level of fluoride in the low-fluoride group (3.30 ± 1.18) mg/L and in the high-fluoride group (5.10 ± 0.35) were observed after 3 months exposure (F = 3.18, P < 0.05). Moreover, the urinary level of fluoride in the low-fluoride group (4.16 ± 1.39) mg/L and in the high-fluoride group (5.70 ± 1.70) mg/L were also observed after 6 months exposure (F = 3.17, P < 0.05). The normal mitochondrial morphology of neurons in rats without fluorosis was observed after 3 and 6 months, while the abnormal mitochondrial morphology of neurons with fluorosis was shown, presenting mitochondrial fragmentation with swollen cristae and even the fragmented, shortened or stacked punctuate membranes (section observation of three bullous mitochondrial-mitochondrial fission process) by TEM. As compared with controls (53.0 ± 4.54 and 1.21 ± 0.18) at the experiment period of 3 months, Mif1 protein analysis with immunocytochemical (the numbers of positive cells: 51.09 ± 6.25) and western-blotting (1.22 ± 0.26) were no significant difference for low fluoride group (t = 1.7, 1.1, P > 0.05); Mif1 protein analysis with immunocytochemical (the numbers of positive cells: 59.71 ± 5.64) and western-blotting (1.66 ± 0.20) were significantly increasing for high fluoride group (t = 2.1, 2.1, P < 0.05). As compared with controls (36.43 ± 4.04 and 1.00 ± 0.13) at the experiment period of 6 months, Mif1 protein analysis with immunocytochemical (the numbers of positive cells 20.05 ± 4.55 and 17.10 ± 3.86) and western-blotting (0.64 ± 0.08 and 0.39 ± 0.06) were significantly decreasing for the two fluoride group (t = 2.1, 2.2; 2.2, 2.2 respectively, all P value were < 0.05).</p><p><b>CONCLUSIONS</b>Taking excessive amount of fluoride might result in the mitochondrial fragmentation for the changed expression of Mfn1, and the neurons damage from the chronic fluorosis might be associated with the dysfunction of mitochondrial fusion.</p>

Influence of chronic fluorosis on the expression of mitochondrial fission protein dynamin-related 1 in the cortical neurons of rats / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the changes of protein expression of mitochondrial fission gene dynamin-related 1(Drp 1) in the cortical neurons of rats with chronic fluorosis.</p><p><b>METHODS</b>A total of 120 one-month-old SD rats (each weighing approximately 100-120 g at the beginning of the experiment) were randomly divided into three groups, and fed with the different doses of fluoride containing in drinking water (untreated control containing 0 mg/L fluoride, and low-fluoride & high-fluoride supplemented with 10 and 50 mg/L fluoride,respectively). After 3 or 6 months exposure, 20 rats from each group were killed. Then the protein expression of mitochondrial fission gene, Drp1, was detected by immunohistochemistry and western-blotting method.</p><p><b>RESULTS</b>Dental fluorosis and urinary fluorosis were obviously found in the rats exposed to fluoride. At the experiment period of 3 months, the numbers of positive cells of Drp1 detected by immunohistochemistry changed. Compared with the control group (36.3 ± 5.8), the changes in low-fluoride group (34.7 ± 4.1) showed no significant difference (t = 1.5, P > 0.05),but the increase in high-fluoride group (45.0 ± 4.7) had statistical significance (t = 8.8, P < 0.05). The western-blotting method had consistent results. Compared with the control group (0.59 ± 0.03), a significant increase of the average topical density in low- fluoride (0.62 ± 0.03) and high-fluoride (0.71 ± 0.02) groups were found (t = 0.02,0.11, P < 0.05). At the experiment period of 6 months, the numbers of positive cells of Drp1 detected by immunohistochemistry significantly changed. Compared with the control group (33.2 ± 4.4), the number in low- fluoride and high-fluoride groups were separately (36.6 ± 3.8) and (39.4 ± 4.2),both increased significantly (t = 3.5,6.3, P < 0.05). Same results could be found in western-blotting method,compared with the control group (0.65 ± 0.06), the average topical density in low- fluoride (0.80 ± 0.09) and high-fluoride (0.76 ± 0.08) groups both increased significantly (t = 0.1,0.1, P < 0.05).</p><p><b>CONCLUSIONS</b>Taking excessive amount of fluoride might result in the changes of expression of Drp1, and the neurons damage from the chronic fluorosis might be associated with the hyperfunction of mitochondrial fusion.</p>

Changed transcription level of mitochondrial fission and fusion gene loci in cortical neurons of rats with chronic fluorosis / 中国地方病学杂志

ObjectiveTo investigate the transcriptional changes of nitochondria fission and fusion gene loci and reactive oxygen species (ROS) level in cortical neurons of rats with chronic fluorosis,and to reveal their roles in mitochondria damage due to chronic fluorosis.MethodsSD rats were fed with different doses of fluoride through drinking water[＜ 0.5(control),10,50 mg/L,respectively] for 3 and 6 months.The level of ROS and mRNA contents of mitochondria fission gene loci Drp1/Fis1 and fusion gene locus Mfn1 in the cortical neurons of rat brains were detected with ROS fluorescent probe and real-time PCR,respectively.ResultsAs compared with control group [10.43 ± 5.98,(3.4 ± 0.6) × 103,(8.8 ± 1.4) × 10,(1.2 ± 0.2) × 102] at the experiment period of 3 months,the level of ROS and mRNA contents of mitochondria fusion gene locus Mfn1 and fission gene loci Drp1/Fis1 in the cortical neurons were obviously increased in the rats fed with 50 mg/L fluoride through drinking water[25.48 ± 6.09,(1.0 ± 0.2) × 1011,(3.0 ± 1.6) × 103,(8.9 ± 3.6) × 102,all P ＜ 0.05],whereas no significant changes were found in the rats fed with 10 mg/L fluoride[11.67 ± 3.49,(3.1 ± 0.3) × 104,(6.7 ± 2.7) × 10,(5.0 ± 0.9) × 10,all P ＞0.05].Furthermore,at 6 months of the experiment the increases in ROS level(63.02 ± 8.15,65.60 ± 7.40) and mRNA contents of mitochondria fission gene loci Drp1/Fis1 [(2.0 ± 0.8) × 106,(4.0 ± 0.6) × 105,(3.8 ± 1.3) × 103,(1.3 ± 0.2) × 103] and the decrease in mitochondrial fusion gene locus Mfn1[(3.0 ± 0.4) × 106、(4.0 ± 0.9) × 104]were observed in the cortical neurons of the rats fed with 10 mg/L and 50 mg/L fluoride as compared with the control group[25.26 ± 6.41,(3.0 ± 0.8) × 109,(5.1 ± 0.8) × 103,(2.8 ± 0.7) × 102,all P ＜ 0.05].Conclusions Excessive intake of fluorine leads to elevated ROS levels,and decreased transcription of mitochondrial fusion gene loci Mfn1,which is positively correlated with the time and dose-exposed to fluoride.The changes of mitochondrial fission and fusion gene loci in the cortical neurons may be related to high level of oxidative stress induced by chronic fluorosis.

Changes of P-glycoprotein and nuclear factor κB in the cerebral cortex of rat with chronic fluorosis / 中国地方病学杂志

Objective The aim of the study was to investigate the expression of P-glycoprotein (P-gp)and nuclear factor κB (NF-κB) in the cerebral cortex of rat with chronic fluorosis,and to reveal the mechanisms of damaged nervous system resulted from the toxicity of fluoride.Methods Sixty SD rats were randomly divided into three groups.The rats in each group were given drinking water containing different levels of fluoride:control group less than 0.5 mg/L,small amount of fluoride exposure group 10.0 mg/L and large amount of fluoride exposure group 50.0 mg/L.The animals were examined at the sixth month after initiating the experiment.Protein levels of P-gp and NF-κB in brain tissues were detected by immunocytochemistry and Western blotting,and the P-gp protein and mRNA level by quantitative real time PCR method.Results As compared to the control group(28.21 ±6.13),the numbers of positive staining cells by P-gp antibody in the cortex of rat brains were significantly increased in both the small and the large amount of fluoride exposure groups[(48.46 ± 8.00),(53.72 ± 9.15),respectively,all P ＜ 0.05] ; the protein levels in the control group(100.00 ± 3.86)％ detected by Western blotting were significantly increased in the cortex of rat brains treated with fluoride in both the small and the large amount of fluoride exposure groups[(189.47 ± 3.14)％,(191.36 ± 11.09)％,respectively,all P ＜ 0.05].The significantly increased expression of NF-κB at the protein level was observed in the cortex of rat brains of the small and the large amount of fluoride exposure groups[(365.97 ± 6.04)％ and (417.15 ± 10.89)％,respectively] as compared with the control group[(100.00 ± 10.07)％,all P ＜ 0.05].The mRMA level of P-gp in the cortex of rat brains of the small and the large amount of fluoride exposure groups(2396 ± 427,3479 ± 371,respectively) were higher than that of the control group(260 ± 106,all P ＜ 0.05).Conclusion The increased expressions of P-gp and NF-κB in the cortex of rat brains are induced by chronic fluorosis,which might be connected with the mechanism of brain damages.

Alteration of mitochondrial distribution and gene expression of fission 1 protein in cortical neurons of rats with chronic fluorosis / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the changes of mitochondrial distribution in axon/soma and the expression of mitochondrial fission 1 (Fis1) protein in the cortical neurons of rats with chronic fluorosis.</p><p><b>METHODS</b>Sixty SD rats were divided into 3 groups (20 each) according to weight hierarchy and fed with different concentrations of fluoride in drinking water (0, 10 and 50 mg/L, respectively) for 6 months. Images of mitochondria and tubulin labeled by immunofluorescence COXIV and tubulin-α were captured with fluorescence microscope. Fis1 protein expression in cortical neurons was analyzed with immunohistochemistry and Western blot. The expression of Fis1 mRNA was detected with real-time PCR.</p><p><b>RESULTS</b>Varying degrees of dental fluorosis and increased fluoride contents in urine were observed in the rats receiving additional fluoride in drinking water. In the cortical neurons of rats fed with 10 mg/L and 50 mg/L fluoride, the numbers of neuronal soma stained with COXIV(34.8 ± 4.7 and 39.3 ± 3.0, respectively), and the expression of Fis1 protein (immunohistochemistry: 54.0 ± 3.6 and 51.3 ± 4.1, respectively; Western blot: 2.9 ± 0.4 and 2.6 ± 0.6, respectively) and mRNA (3773 ± 1292 and 1274 ± 162, respectively) was markedly increased as compared with controls (4.4 ± 2.3, 25.2 ± 2.5, 1.8 ± 0.2 and 277 ± 73) over the experimental period of 6 months.</p><p><b>CONCLUSIONS</b>Excessive intake of fluoride results in an altered mitochondrial distribution in axon and soma in cortical neurons (i.e., the increase in soma and the decrease in axon), increased expression of Fis1 gene and enhanced mitochondrial fission. The altered mitochondrial distribution may be related to the high expression level of Fis1 and a functional disorder of mitochondria.</p>

Changes of reactive oxygen species level and mitochondria fission-fusion hi cortical neurons of rats with chronic fluorosis / 中国地方病学杂志

Objective To investigate the changes of reactive oxygen species(ROS) level and mitochondria fission-fusion-balance in cortical neurons of rats with chronic fluorosis and reveal the correlation between these two factors. Methods One hundred and twenty rats were randomly divided into 3 groups(control group, low-dose fluorosis group, high-dose fluorosis group) and 40 rats were in each group according to body weight and the experiments were carried out for 3 months or 6 months. The rats were fed with different concentrations of fluoride (NaF) to establish fluorosis models. Controls were fed with tap water( < 0.5 mg/L), experimental animals in low- or high-dose group were fed with water containing NaF 10.0,50.0 mg/L, respectively. The level of ROS and the morphology in mitochondria fission-fusion balance in neurons of the cortex of rat brains prepared with cortical frozen sections were detected with ROS fluorescent probe and MitoTracker RED probe, respectively. Results Significant differences of the level of ROS and the numbers of abnormal mitochondria in morphology in the cortical neurons were found between 3 groups at the experiment period of 3 month and 6 month(F= 3.07,3.06,3.05,3.07, all P < 0.05). As compared with control group(10.43 ± 5.98,4.12 ± 3.86) at the experiment period of 3 month, the level of ROS and the numbers of abnormal mitochondria in morphology in the cortical neurons were obviously increased in high-dose fluorosis group(25.48 ± 6.09,20.47 ± 6.09, all P < 0.05), whereas no significant changes were found in low-dose fluorosis group(11.67 ± 3.49,6.68 ± 3.48, all P> 0.05). Furthermore, the increases in both ROS level and abnormal numbers of mitochondria were significant observed in the cortical neurons of low-dose fluorosis group (63.02 ± 8.15, 49.33 ± 8.61) and high-dose fluorosis group(65.60 ± 7.40,53.10 ± 6.95) as compared with the control group (25.26 ± 6.41,20.26 ± 6.41) at the experimental period of 6 month (all P < 0.05). The abnormal numbers of mitochondria correlated with ROS level(r = 0.93,0.81, all P < 0.05). Conclusions Taking excessive amount of fluoride results in high level of oxidative stress and impaired the balance of mitochondrial fission-fusion,which is dependent on the feeding times and doses of fluoride. The mechanism of the mitochondrial abnormalities might be associated with the high level of oxidative stress induced by chronic fluorosis.

A reviewing for profiling of highly degraded remains / 法医学杂志

It's always a challenge to type from highly degraded biological remains. PCR-based STR genotyping is helpful and valuable for such degraded samples like bones, teeth et al, but the typing results are sometimes unstable or wrong. Here the methods for solving the problems and improving reproducibility are reviewed.

Polymorphism of DXYS267 locus and application of its Y-specific single nucleotide substitutions / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the polymorphism of DXYS267 locus in China Han population and find the application and characters of its Y-specific single nucleotide substitutions.</p><p><b>METHODS</b>The locus was analyzed by PCR and PAGE in silver-staining. The Y-specific STR was amplified with newly designed primers according to the Y-specific single nucleotide substitutions.</p><p><b>RESULTS</b>Six alleles were detected in Han population in Wuhan. Exact tests demonstrated that genotype frequencies did not deviate from Hardy-Weinberg equilibrium. Heterozygosity of DXYS267 was 0.6706, discrimination power (DP) was 0.8433, and the probability of paternity exclusion (PE) was 0.5957. The Y-specific STR of DXYS267 was successfully amplified with the new primer. The 4 alleles for Y-STR were detected with haplotype diversity (HD) 0.6372.</p><p><b>CONCLUSION</b>The DXYS267 locus is appropriate for individual identification and paternity testing. The new primer is useful for individual and paternity testing involving brothers and mixed stains.</p>

Application of the age-associated injure in mitochondrial DNA / 法医学杂志

Nowadays, the injury in human mitochondrial DNA (mtDNA) is well known to accumulate in various tissues with age. It's significant to further investigate and then apply it to estimation of the age at parenchymas.