Study on the Specific Complexation of GMDTC and Metal Ion / 中华劳动卫生职业病杂志

Objective@#Study the response of GMDTC to cadmium ions and metal ions in vivo to determine whether GMDTC are specifically complexed with cadmium ions to provide a reference for the safety and dfficacy of GMDTC.@*Methods@#Complexometric titration, HPLC and HPLC-MS were applied to research the complexation reaction of GMDTC and various metal ions. The molecular ion peak of GMDTC, GMDTC-Cd complex and GMDTC-Pb complex also detected by LC-MS. Additionally, the initial structure was determined by DFT simulation method.@*Results@#Results of complexometric titration and HPLC detection showed that GMDTC characteristic absorption peak area was proportional to the concentration of itself and there was no color change and peak time change when the GMDTC mixed with Ca2+, Fe2+, Mg2+, Zn2+. However, the color changed to black transition when the GMDTC mixed with Cu2+ and the color changed from yellow precipitate to light yellow transparent transition when GMDTC mix with Hg2+. Moreover, the peak area as well as the retention time has changed a lot which indicated that a chemical reaction has already happened. When the GMDTC mixed with Cd2+ and Pb2+, the color has changed from pale yellow to colorless transparent and the peak area of GMDTC has increased a lot. Finally, the GMDTC-Cd complex ratio both of which are 2:1 were calculated based on the results of LC-MS instrument and atomic calculations.@*Conclusion@#The specific cadmium chelating agent GMDTC can not react with the Ca2+, Fe2+, Mg2+, Zn2+, but it can react chemically with Cu2+ and Hg2+, even specific complex with Pb2+ and Cd2+.

Exploration of the immune mechanism in systemic lupus erythematosus MRL/lpr mice / 中国比较医学杂志

Objective To explore the immune mechanism in the systemic lupus erythematosus MRL/lpr mice at different months of age, and to provide the basis for research of its pathogenesis. Methods 3-,4-,5- and 6-month old female MRL/lpr mice, and wild type C57 female mice were used in this study, 10 mice per each group. Their organ coefficients were determined. ELISA was performed to detect the serum levels of double stranded DNA(ds-DNA) antibody. The interleukins IL-2,IL-4,IL-17 and tumor necrosis factor-α(TNF-α)in spleen tissue were detected. Flow cytometry was used to assess the content of spleen lymphocyte CD3 cells and CD4/CD8 cell ratio. Results Compared with the 3-month old wild-type C57 mice,the spleen coefficient,the blood concentration of ds-DNA antibody,IL-2 and TNF-α in the 3- to 6-month old MRL/lpr mice were significantly increased(P < 0.05). There was no significant difference between the concentrations of interleukin IL-4(P> 0.05). The blood concentration of IL-17 in the 5- and 6-month old MRL/lpr mice was significantly lower(P < 0.05 or P < 0.01)than that in the 3-month old MRL/lpr mice. The rest indexes of MRL/lpr mice showed no obvious changes or significant difference in the mice at different ages. Compared with the 3-month old C57 mice,the spleen CD3 lymphocyte concentration in the MRL/lpr mice was significantly decreased(P< 0.01). With the increasing age, the CD3 lymphocyte concentration and D4 +/CD8 +cell ratio in the MRL/lpr mice were decreased, however, showing a non-significant difference(P ﹥0.05). Conclusions The data obtained in this study indicate that 3-month old lupus MRL/lpr mice have already immune injury,increasing with the increase of age of the mice.

New Zealand rabbit model of cisplatin-induced acute kidney injury / 中国职业医学

OBJECTIVE: To establish a New Zealand rabbit model of acute kidney injury induced by cisplatin. METHODS: A total of 24 male New Zealand rabbits were randomly divided into control group,low-,medium-and high-dose cisplatin group according to the body mass. Rabbits were injected with cisplatin at 0. 0,1. 0,2. 0,4. 0 mg/kg body weight by auricular vein. Rabbits in low-dose group was continuously injected for 5 days,medium-dose group was continuously injected for 3 days,and the high-dose group was injected for once per day. Rabbits in the control group did not receive any treatment. Blood was collected from the middle ear artery and 24 h urine was taken before exposure and on day 1,day 3,day 5 and day 7 of injection. The serum creatinine( Scr) and urea nitrogen( BUN) were detected by colorimetric method,and 24 h urine kidney injury molecule 1( KIM-1) was measured by enzyme-linked immunosorbent assay. Plasma platinum,24 h urinary platinum and renal platinum level were detected by inductively coupled plasma mass spectrometry.At the end of the experiment,rabbits were sacrificed and the left kidney was taken for histopathological examination.RESULTS: The body mass of rabbits of the low-,medium-and high-dose groups on day 7 after cisplatin exposure was lower than that of the control group( P < 0. 05),and lower than that of the same group before exposure( P < 0. 05). After 3 days of exposure,the Scr level in each dose group was higher than that of the control group( P < 0. 05),the Scr level on day 3and day 5 in medium-and high-dose groups were higher than that of the low-dose group( P < 0. 05). The BUN levels on day 3 and day 5 in medium-and high-dose group were higher than that of the control group and low-dose group( P <0. 05),the BUN levels on day 7 in three dose groups were higher than that of the control group( P < 0. 05). The levels of plasma platinum and 24 h urinary platinum in the three doses groups of New Zealand rabbits on day 1,day 3,day 5 and day 7 after exposure were higher than that of the control group( P < 0. 05),and were higher than the pre-treatment levels of the same group( P < 0. 05). The level of 24 h urinary KIM-1 in the meclium-dose group of New Zealand rabbits was higher than that of the control group on day 3 of exposure( P < 0. 05). The level of 24 h urinary KIM-1 in the mediumdose group of New Zealand rabbits on the 5th day after exposure was higher than that of the control group( P < 0. 05). The renal platinum levels in the three groups of New Zealand rabbits were higher than that in the control group( P < 0. 05).The pathological changes of rabbit kidney caused by cisplatin are mainly tubular dilatation,protein cast,alkalophilic and interstitial nephritis. CONCLUSION: Cisplatin can induce acute kidney injury in rabbits,and the degree of injury is dosedependent. The dose of 1. 0 mg/kg body weight continuous injection for 5 days is closely related to clinical use of cisplatin,which is recommended for model establishment.

Study on the dosage of GMDTC for removing cadmium / 中国职业医学

OBJECTIVE: To explore different doses of sodium(s)-2-(dithiocarboxylato((2R,3R,4R,5R,6R)-2,3,4,5,6-pentahydroxyhexyl) amino)-4-(methylthio) butanoate(GMDTC) for removing cadmium. METHODS: Thirty-five male New Zealand rabbits were randomly divided into blank control group,GMDTC high dose control group,model control group,ethylene diamine tetraacetic acid(EDTA) control group and GMDTC low,medium and high dose groups,five rabbits in each group. The blank control group and GMDTC high dose control group were given 0. 90% normal saline solution intravenously; model control group,EDTA control group and GMDTC low,medium and high dose group were given 2 μmol/kg of cadmium chloride(CdCl_2) and 40 μmol/kg of β-mercaptoethanol mixed solution intravenously,5. 0mL/kg body weight(bw),once a day for five days. On the forty-one day of the experiment(the fist day of GMDTC treatment),the control group and the model control group were injected 0. 90% normal saline solution 250 mL via ear vein,the EDTA control group was given EDTA solution at the dose of 93. 5 mg/kg bw with 250 mL 0. 90% normal saline solution,also via ear vein; the GMDTC high dose control group,and the GMDTC low,medium and high dose groups were given 250 mL GMDTC solution at the concentration of 108.0,12.0,36.0 and 108. 0 mg/kg bw with 0. 90% normal saline by intravenous infusion,once a day,6 times a week for four consecutive weeks. The urine β_2-microglobulin(MG),renal cadmium,blood cadmium,and urinary cadmium before and after the treatment were detected. RESULTS: The body weight of New Zealand rabbits increased with the increasing feed time(P < 0. 01). The levels of β_2-MG before treatment increased in model control group,EDTA control group,GMDTC low,medium and high dose groups than that in the blank control group(P < 0. 01). The levels of renal cadmium after treatment in GMDTC medium and high dose groups decreased compared with those in the blank control group and EDTA control group respectively(P < 0. 05). The blood cadmium after treatment in EDTA control group,GMDTC low,medium and high dose groups were decreased compared with those before treatment in the same group respectively(P < 0. 05),meanwhile decreased than the blood cadmium after treatment in the model control group respectively(P < 0. 05). The blood cadmium after treatment had not a statistically significant difference among the EDTA control group,GMDTC medium and high dose groups(P < 0. 05). At all the time points(1,6,8,13,15,20,22 and 28 days after treatment),the urinary cadmium after treatment in EDTA control group and the three GMDTC dose groups increased compared to the model control group at the same time(P < 0. 05). The urinary cadmium after treatment increased with GMDTC dose increased at the other six time points,expect on 20 and 22 days after treatment(P < 0. 05). The blood cadmium removal rates after treatment were 70. 06%,74. 86% and 78. 05% and the renal cadmium removal rates were 14. 27%,27. 95% and 61. 24% in GMDTC low,medium and high dose groups,respectively. CONCLUSION: The intravenous infusion of GMDTC at the dose of 108. 0 mg/kg bw effectively removed cadmium in cadmium poisoning rabbit. This dose had no obvious toxic effect and was equivalent to human dose of 36. 0mg/kg bw which meets the requirement of new drug property.

The influencing factors for establishing a model of acute myocardial infarction in rats / 西安交通大学学报(医学版)

ABSTRACT:Objective To study the effects of weight and anteroposterior diameter-to-transverse diameter ratio on establishing a model of acute myocardial infarction (AMI)in rats without artificial ventilation and changes in left ventricular function after infarction.Methods Healthy SD rats were randomly divided into group A (200-250 g),group B (250 - 300 g),group C (> 300 g),and group D (control group).The left anterior descending (LAD)coronary artery was ligated to establish a model of myocardial infarction under spontaneous breathing condition immediately after thoracic lines were measured.And changes of electrocardiography were recorded after model establishment.At 2 and 4 weeks after AMI,we observed ventricular wall thickness and ventricular wall motion and measured the changes of cardiac function.Histomorphological changes and myocardial ultrastructure of the heart were observed under thoracotomy 2 weeks after operation.The above data were analyzed by SPSS13.0 statistics software.Results ① The first AMI rat model was established successfully after 30 times of experiments, and after 100 times the model’s success rate gradually stabilized at about 83%.② Group B and group C had a higher model success rate than group A (P 0.05).③ There was no association between the rate of rat thoracic line and modeling success rate (P >0.05). ④ Two weeks after thoracotomy,ischemic myocardial color was white,and ventricular wall motion decreased.HE staining revealed that cardiomyocytes disappeared and were replaced by fibrous tissues and collagen.Remnant cardiomyocytes were arranged disorderly and myofibers were fractured,with interstitial damage and hyperplasia of fibrous tissue.Visible muscle cells were sparse and dissolved,the mitochondria had darker staining,blurred cristae, and edema under electron microscopy. ⑤ Compared with group D, 2 weeks and 4 weeks after myocardial infarction,left ventricular end-diastolic diameter (LVEDD)and left ventricular end systolic diameter (LVEDs) increased (P <0.05),but EF values and heart rate dropped (P <0.05).Conclusion By this method,a model of AMI in rats can be established successfully and the heart function is changed.Under the condition of non-artificial ventilation,the weight of rats is an important factor for establishing AMI model.However,we have not confirmed the effect of thoracic lines on establishing AMI model yet.

Breeding management and quality control of SPF BALB/c nude mice / 中国比较医学杂志

Breeding and quality control of SPF nude mice involves many links and works .The quality and quantity of animals are directly affected by breeding management and quality control .A standardized breeding system of SPF BALB/c nude mice were gradually established at the center after 2007.This paper introduces specific practices and experience in facilities, stock, breeding management , and quality control of SPF BALB/c nude mice, in order to provide the reference for related breeding and study in SPF BALB/c nude mice.

Brief report of series study on Rongshui miniature pig as laboratory animal / 中国比较医学杂志

The provenances of Rongshui miniature pigs ( RMPs) were purchased from Rongshui, Guangxi, in 2012.130 RMPs were transported to Sanshui, Guangdong,China, which were breed according to the laboratory animal standards.83 RMPs were selected randomly from the first filial generations ( F1 ).The basic data were collected including breed characteristics, reproductive performance, grow curve, hematology, biochemical markers of serum and urine, organ coefficient, Chromosome analysis.According to the national and local standards, the quality control standards of RMP were set up including microbiological, parasitic, environment and facilities, fodder, pathology, genetic.The results showed that RMPs adapt to the climate of Guangdong.The natural mating and conceive rate was 88.3% with the pregnancy of 112 days.The average number of firstborn and still-born was 6.1 and 7.9 respectively.RMP was small body size with the adult body weight of 17.21 ±5.20 kg and 16.35 ±5.23 kg in female and male respectively.RMP was very tame.The mitochondrial genome analysis suggested RMP belonged a typical miniature pig breed in China, which is ancient than Lanyu pig.RMP could be breed as a new kind of laboratory animal.

Genomic and phylogenetic analysis of porcine mitochondrial genomes of Rongshui miniature pig / 中国比较医学杂志

Objective Assembly whole mitochondrial genome sequence of Rongshui miniature pig ( RMP ) breed and analysis the structure of mitochondrion based on the next-generation sequecing method.Comparison of phylogenetic relationship and genetic diversity among different pig breeds.Methods We collected peripheral venous blood sample from RMP and constructed two paired-end sequencing libraries.A whole-genome shotgun sequencing strategy and Illumina Genome Analyser sequencing technology were used in our study.Results The mitochondrial genome of RMP consists of 13 protein coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes and the length of pig is 16888 bp.The GC content of this pig mitochondrial genome is about 44 %.Based on phlogenetic analysis, population genetic analysis, our findings confirmed that the ancestral cluster in East Asia mainly occurred among Diannan 7#pig, Hainan wild boar, Lanyu and RMP.Conclusion RMP, a typical miniature pig breed in China, is an earlier ancestor than Lanyu pig breed.

Detection and analysis of blood parameter, blood biochemical and electrolyte indices in F1 generation of Rongshui miniature pig / 中国比较医学杂志

Objective To detect the blood parameter, blood biochemical and electrolyte indices of the F1 generation of Rongshui miniature pig ( RMP) .Methods The blood of 43 female and 42 male RMPs of 4 th month old, and 36 RMPs of 12th month old ( half male and female) were extracted from jugular vein.And the blood parameter, blood biochemical and electrolyte indices were detected by blood analyzer and automatic biochemical analyzer.Results In the same month-old RMP, no significant difference between male and female were found in most indices of blood parameter, blood biochemical and electrolyte indices.On the other hand, many indices were difference between 4th month old and 12th month old RMPs of same gender.Compared with the 4th month old RMP, the 12th month old RMP decreased significantly in WBC and PLT, increased in HGB ( P 0.05 ) .Serum ALT, AST, ALP, CK (male), LDH(male), A/G, BUN, GLU (female), CHOL (male) and K+decreased significantly (P <0.05), while serum TP, TBIL, CR and Ca2+increased significantly (P <0.05),but serum CHOL, TG, HDL-C and LDL-C were not different.86.4%(19/22) biochemical and electrolyte indices in RMP were in/or close to the range of normal value of human.Conclusion Most of the blood parameter, blood biochemical and electrolyte indices of RMP were close to human’ s normal value.

Autopsy procedures and pathological observation of spontaneous main organ lesions in Rongshui miniature pig / 中国比较医学杂志

Objective To establish the data including anatomy and histology of main organs in Rongshui miniature pig (RMP).Methods F1 Rongshui miniature pigs with male and female (2 in each group) in 6 month old were used in this experiment.We measured body weights, dissected these pigs after anaesthesia, recorded total blood volume, total plasma volume, number of spine and dental formula, took main organs for photographs, and made histological sections observed and took photographs by microscope.Results We gained the photographs of main organs and histological sections, organ weights,organic coefficients and other basic data.Conclusion Basic anatomy and histology data of main organs in RMP were collected.

Correlation between expression of TOP2A and HER2 signaling pathway in breast cancer / 西安交通大学学报(医学版)

Objective To study the correlation of expression of DNA topoisomerase Ⅱ alpha (TOP2A)with expressions of human epidermal growth factor receptor 2 (HER2)and phosphatase and tensin homolog (PTEN)and gene mutation of phosphatidylinositol 3-kinase (PI3K)in breast cancer so as to provide reference for prognosis of the cancer and evaluation of drug efficiency.Methods This study enrolled totally 96 breast cancer patients. Tumor specimens were resected.The gene expressions of TOP2A,HER2 and PTEN were analyzed using branched DNA-liquid-chip,and PI3K gene mutation was detected by xTAG-liquid-chip.Correlations between gene expressions and gene mutation were further explored by Spearman correlation analysis so as to clarify the relationship between TOP2A and HER2 signaling pathway gene.Results Co-expression of TOP2A and HER2 was strong,and TOP2A tended to be highly expressed in the presence of high expression of HER2 (P =0.01).The expression of PTEN was not significantly correlated with the expression of TOP2A,whereas the mutation of PI3K had a positive association with the high expression of TOP2A (P =0.004).Conclusion Anthracycline drug resistance factor TOP2A may be related to the critical factors of HER2 signaling pathway,suggesting that HER2 expression and PI3K mutation may be key factors in regulation of TOP2A expression,which would provide important evidence for chemotherapeutic resistance.

Morphological characterization of the rat model of acetic acid-induced acute rectal mucosal injury / 中国实验动物学报

Objective To establish a reproducible rat model of acute rectal mucosal injury induced by acetic acid. Methods Fifteen healthy Sprague-Dawley rats were randomly divided into the control group (3 rats) and experimental group (12 rats).Acute rectal mucosal injury was induced by 4%acetic acid using a cotton swab inserted into the rat rec-tum for 1 min to a depth of 3 cm.The morphological characteristics were analyzed by the naked eye and histology at 0.5 h and 1, 4, and 6 days after acetic acid intervention.Results All rats survived 6-day study period.The successful rate of model establishment was 100%.From 0.5 h to 1st day after acetic acid intervention, the gross morphology of recta showed congestion, edema and ulcer to ulcer complicated with hemorrhage.The histology showed necrosis and hemorrhage of the epithelial tissue of the mucosa to complete and extensive necrosis of the mucosa.The glandular structure showed partial to complete loss.The submucosa showed edema to edema complicated with hemorrhage and congestion.The interstitial tissues showed vasodilatation and congestion to inflammatory cell invasion.From 4 to 6 days after acetic acid intervention, the rectal mucosal changes were obviously improved.Epithelial and glandular regeneration and inflammatory granulation occurred, but not fully recovered, some edema and redness, partial lack of glands were still present.Conclusions 4%acetic acid for 1 min can be used to successfully induce rat model of acute rectal mucosal injury.This procedure is easy to operate, with a high success rate,reproducible, and the alterations are lasting more than 6 days.This animal model is very suitable for rapid screening of topical drugs for the treatment for rectal mucosal injury.

Effect of Rhodiola Crenulate oral liquid on reproduction function in male rats / 国际中医中药杂志

Objective To observe the action of Rhodiola Crenulate(RC) oral liquid reproduction function for the male rats. Methods 100 SD rats were randomly divided into 5 groups with equal gender number respectively. The male rats were given RC oral liquid (0, 2.48 g/kg, 7.48 g/kg, 24.80 g/kg) and Nanbao (2.00 g/kg)by oral twice a day for 48 d, respectively. Then the male rat was put into the same box in which the female rat whit two-side ovarys were excised for 15 d, respectively. The intercoursing ability,testosterone levels, and the weight of immune organs of the male rats were examined. 40 four-week-old rats were divided into 5 groups. Except for blank control, all rats's testis were excised. The castrated rats were treated with RC oral liquid (2.48 g/kg, 24.8 g/kg)and Methyltestosterone (2.00 mg/kg). The blank control group and model control group were given the same dose NS by oral twice a day for 28 d, respectively. The weight of accessory sex organs in castrated rats was observed. Results The level of testosterone and the organ coefficients of thymus and spleen in RC oral liquid groups were higher than blank control (P<0.05 or P<0.01).The weight of accessory sex organs of castrated rats in RC oral liquid groups were heavier than model control (P<0.05). Conclusion RC oral liquid can improve the reproduction function in male rats.

Effect of fluoroacetamide on cardiomyocytes of rat and the antidotal effect of acetamide / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To observe the effect of fluoroacetamide on cardiomyocytes of rat and the antidotal effect of acetamide.</p><p><b>METHODS</b>4 groups of SD rats were treated with various dosages of fluoroacetamid(p.o.) and 2 groups of them were treated with acetamide(i.p.). The changes of cardiomyocytes and serum AST, LDH, CK, CK-MB and HBDH were measured at different intervals after poisoning.</p><p><b>RESULTS</b>In the group treated with fluoroacetamid 8 mg/kg. bw, serum AST[(589.58 +/- 821.72) U/L], CK[(916.78 +/- 343.55) U/L], HBDH[(504.47 +/- 148.88) U/L] raised obviously compared with control[(187.70 +/- 46.87), (755.65 +/- 498.90), (347.25 +/- 228.40) U/L respectively] (P < 0.01), and the pathological findings such as degeneration, liquefactive necrosis and filtration of inflammatory cells in cardiac muscles were observed 24 hours later, while all the male dead within 3 days. In the group treated with fluoroacetamid 4 mg/kg. bw, serum LDH and HBDH rose significantly compared with control(P < 0.01) 5 day later. On the day of 10, myocardial enzymes restored in all experiment groups with some interstitial fibroblastic proliferation. The pathological changes were reduced in the group treated with acetamide synchronously (100 mg/kg. bw).</p><p><b>CONCLUSION</b>Acute intoxication of fluoroacetamide could damage cardiomyocytes while acetamide could reduce the injury of them, but the injury was reversible. The levels of serum myocardial enzymes could be a usable index for early diagnosis.</p>

Construction of recombinant vector containing fusion gene NT_4-Ant-Shepherdin[79-87] / 西安交通大学学报(医学版)

Objective To investigate survivin as an anticancer therapeutic target by use of shepherdin [79-87],a novel peptide carrying the survivin sequence from Lys-79 through Leu-87,we constructed an recombinant vector containing fusion gene NT4-Ant-Shepherdin [79-87].Methods The gene of Ant-Shepherdin [79-87] was obtained by PCR and T-vector method.After cloned and digested with restricted enzyme,Ant-shepherdin [79-87] was inserted in PBV220NT4 vector.The recombinant vector was transformed into the competent cell,E.coli DH5?.The fusion gene of NT4-Ant-Shepherdin [79-87] was identified by agarose gel electrophoresis (AGE).Results DNA sequencing results verified that the sequence of Ant-Shepherdin [79-87] was consistent with what we had designed.After transformed E.coli DH5?,a fragment of 321 bp was confirmed.Conclusion The recombinant vector containing fusion gene NT4-Ant-Shepherdin [79-87] was successfully constructed in this experiment by molecular biology techniques,which provides the basis of further research of survivin for cancer gene therapy.