ABSTRACT
Objective To evaluate the protective effect of recombinant Schistosoma japonicum cystatin (rSj-Cys) against acute kidney injury induced by acute liver failure and unravel the underlying mechanism, so as to provide insights into the clinical therapy of acute kidney injury. Methods Twenty-four male C57BL/6J mice at ages of 6 to 8 weeks were randomly divided into the normal control group, rSj-Cys control group, lipopolysaccharide (LPS)/D-galactosamine (D-GaIN) model group and LPS/D-GaIN + rSj-Cys treatment group, of 6 mice each group. Mice in the LPS/D-GaIN group and LPS/D-GaIN + rSj-Cys group were intraperitoneally injected with LPS (10 μg/kg) and D-GaIN (700 mg/kg), and mice in the LPS/D-GaIN + rSj-Cys group were additionally administered with rSj-Cys (1.25 mg/kg) by intraperitoneal injection 30 min post-modeling, while mice in the rSj-Cys group were intraperitoneally injected with rSj-Cys (1.25 mg/kg), and mice in the normal control group were injected with the normal volume of PBS. All mice were sacrificed 6 h post-modeling, and mouse serum and kidney samples were collected. Serum creatinine (Cr) and urea nitrogen (BUN) levels were measured, and the pathological changes of mouse kidney specimens were examined using hematoxylin-eosin (HE) staining. Serum tumor necrosis factor (TNF)-α and interleukin (IL)-6 levels were detected using enzyme-linked immunosorbent assay (ELISA), and the expression of inflammatory factors and pyroptosis-related proteins was quantified in mouse kidney specimens using immunohistochemistry. In addition, the expression of pyroptosis-related proteins and nuclear factor-kappa B (NF-κB) signaling pathway-associated proteins was determined in mouse kidney specimens using Western blotting assay. Results HE staining showed no remarkable abnormality in the mouse kidney structure in the normal control group and the rSj-Cys control group, and renal tubular injury was found in LPS/D-GaIN group, while the renal tubular injury was alleviated in LPS/D-GaIN+rSj-Cys treatment group. There were significant differences in serum levels of Cr (F = 46.33, P < 0.001), BUN (F = 128.60, P < 0.001), TNF-α (F = 102.00, P < 0.001) and IL-6 (F = 202.10, P < 0.001) among the four groups, and lower serum Cr [(85.35 ± 32.05) μmol/L], BUN [(11.90 ± 2.76) mmol/L], TNF-α [(158.27 ± 15.83) pg/mL] and IL-6 levels [(56.72 ± 4.37) pg/mL] were detected in the in LPS/D-GaIN + rSj-Cys group than in the LPS/D-GaIN group (all P values < 0.01). Immunohistochemical staining detected significant differences in TNF-α (F = 24.16, P < 0.001) and IL-10 (F = 15.07, P < 0.01) expression among the four groups, and lower TNF-α [(106.50 ± 16.57)%] and higher IL-10 expression [(91.83 ± 5.23)%] was detected in the LPS/D-GaIN + rSj-Cys group than in the LPS/D-GaIN group (both P values < 0.01). Western blotting and immunohistochemistry detected significant differences in the protein expression of pyroptosis-related proteins NOD-like receptor thermal protein domain associated protein 3 (NLRP3) (F = 24.57 and 30.72, both P values < 0.001), IL-1β (F = 19.24 and 22.59, both P values < 0.001) and IL-18 (F = 16.60 and 19.30, both P values < 0.001) in kidney samples among the four groups, and lower NLRP3, IL-1β and IL-18 expression was quantified in the LPS/D-GaIN + rSj-Cys treatment group than in the LPS/D-GaIN group (P values < 0.05). In addition, there were significant differences in the protein expression of NF-κB signaling pathway-associated proteins p-NF-κB p-P65/NF-κB p65 (F = 71.88, P < 0.001), Toll-like receptor (TLR)-4 (F = 45.49, P < 0.001) and p-IκB/IκB (F = 60.87, P < 0.001) in mouse kidney samples among the four groups, and lower expression of three NF-κB signaling pathway-associated proteins was determined in the LPS/D-GaIN + rSj-Cys treatment group than in the LPS/D-GaIN group (all P values < 0.01). Conclusion rSj-Cys may present a protective effect against acute kidney injury caused by acute liver failure through inhibiting inflammation and pyroptosis and downregulating the NF-κB signaling pathway.
ABSTRACT
Objective To explore the anti-tumor effect of 17XL strains of Plasmodium yoelii(P.y)infection on melanoma in mice. Methods B16F10 tumor cells were axillarilly injected into the right flank of 20 C57BL/6 mice to establish tumor-bearing mouse models. The next day,the mice were randomly divided into a P.y infection group and control group,10 mice each group. Each mouse of the P.y infection group was intraperitoneally injected with 1×106 red blood cells including 20% P.y infection red blood cells,and each one of the control group were intraperitoneally injected with 1×106 normal red blood cells of C57BL/6 mice. The time of tumor formation of the mice in the two groups was observed and the tumor volumes were measured. Results The time of tumor formation in the P.y infection group[(11.30 ± 0.21)d]was significantly later than that in the control group [(10.40 ± 0.22)d](P < 0.05). From the tumors could be accurately measured to the study end point,both the tumors of mice in the two groups were growing,and the tumor volumes of mice in the P.y infection group were significantly less than those in the control group at each time point(all P < 0.05). The growth rate of tumors in the P.y infection group[(71.10 ± 6.29)mm3/d]was significantly slower than that in the control group[(302.80 ± 49.94)mm3/d](P < 0.05),and the growth rates of tumors every day in the P.y infection group were significantly slower than those in the control group(all P < 0.05). Conclusion The P.y in-fection can delay the occurrence of tumor and inhibit the growth of melanoma.
ABSTRACT
Teaching competition is an effective way for college and university teachers to improve their teaching skills. Based on the teaching practice and experience in medical parasitology,this paper discusses several key issues in teaching competition including topics,teaching designs and teaching methods. It provides references for the teachers in department of parasitology of universities and colleges to improve the quality of classroom teaching.
ABSTRACT
Objective To study the effect of exogenous nitric oxide donor sodium nitroprusside(SNP)on antioxidant en?zymes activities and lipid peroxidation of mice infected with Trichinella spiralis. Methods BALB/c mice were infected with T. spiralis separated by the digestion method. Forty?two days post?infection,the peripheral blood and hepatic tissue from the in?fected or normal mice were collected. Then 4 groups were set:liver homogenate from infected mice+SNP(Group A),liver ho?mogenate from normal mice+SNP(Group B),peripheral blood from infected mice+SNP(Group C),and peripheral blood from normal mice+SNP(Group D). The final concentrations of SNP in each group were set as 0(blank control),2,5,10μmol/L and 30μmol/L,respectively. After reacting with SNP at 37℃for 30 min,the superoxide dismutase(SOD),glutathi?one peroxidase(GSH?Px),catalase(CAT)activities,and malondialdehyde(MDA)concentration were examined and com?pared. Results The levels of SOD,CAT,GSH?Px and MDA concentration in the liver and the blood from the mice infected with T. spiralis were significantly higher than those of the normal ones(all P < 0.05). When reacted with 10 μmol/L and 30 μmol/L SNP,the SOD,GSH?Px,and CAT activities in Group A and B decreased significantly(all P<0.05),while the liver MDA concentration reacted with 2-30μmol/L SNP increased obviously(all P<0.05). As reacted with 30μmol/L SNP,the ac?tivities of blood SOD,GSH?Px,and CAT in Group C and D decreased,while the MDA concentration in blood still increased (all P<0.01). When the SNP concentration was in the range of 2-30μmol/L,there were a negative correlation between the SNP concentrations and SOD,GSH?Px,and CAT activities,as well as a positive correlation with the MDA concentration in the liver and blood from the mice infected with T. spiralis(all P<0.05). Conclusions T. spiralis infection could cause oxidative damage to mice,and increase SOD,GSH?Px,and CAT activities. Nitric oxide released from SNP can decrease antioxidase activ?ities,and inhibit the antioxidant capacity of mice infected with T. spiralis.
ABSTRACT
Objective To understand the status of resistance to multiple insecticides and the frequencies of kdr mutations in Culex pipiens pallens from north?central Anhui Province. Methods From July to September,2014,the C. pipiens pallens mosquito larvae were collected in Huaibei,Bengbu and Chuzhou cities of the north?central Anhui Province and reared to adults. The female adult mosquitoes at 3-5 days post emergence were tested for susceptibility to the four insecticides,namely 0.05%deltamethrin,5%malathion,0.1%bendiocarb and 4%DDT,by using the standard WHO resistance tube bioassay. The detec?tion of the point mutations of the kdr gene at codon 1014 was conducted by PCR and DNA sequencing in the deltamethrin?resis?tant and?susceptible mosquitoes. Results High levels of resistance to all the four insecticides were found in all the three tested populations,although mosquito mortality varied among populations and test insecticides. Among the test insecticides,DDT showed lowest mortality with no significant difference(F=1.027,P>0.05)in all test populations,whereas significantly differ?ent mortalities were observed among populations for the remained three insecticides tested(deltamethrin,malathion,and ben?diocarb)(F = 23.823,33.955,128.841;all P < 0.01). Two types of non?synonymous kdr mutation at codon position 1014 (L1014F and L1014S)were observed. A positive correlation between L1014F mutation frequencies and deltamethrin resistance levels were detected in the three mosquito populations(r2=0.718,P<0.01). Conclusions The observed high levels of resis?tance to multiple?insecticides coupled with the occurrence of medium to high kdr frequencies in populations of C. pipiens pallens could profoundly affect the mosquito vector control programme in China. The local health departments need to strengthen vector dynamic monitoring and implement rational resistance management strategies.
ABSTRACT
Objective To observe the effect of excretory/secretory products from Trichinella spiralis adult worms(AES)on cecal ligation and puncture(CLP)?induced sepsis in mice. Methods Forty?eight BALB/c mice were randomly divided into 3 groups:a sham operation group(PBS+sham group,Group A),a CLP?induced sepsis group(PBS+CLP group,Group B)and an AES treatment group(AES+ CLP group,Group C). The mice of each group were intraperitoneally injected with 25 μg of AES or PBS only as a control in a total volume of 200μl. Eight mice from each group were selected randomly for survival analy?sis of 96 hours. The other 8 mice in each group were observed for pathological changes in the lung,liver and kidney tissues by HE staining 12 h after CLP,and then determined for the detection of cytokines including TNF?α,IL?1β,IL?6,IL?10 and TGF? βin the sera by ELISA. Results The difference among the survival rates of mice in the 3 groups was statistically significant (χ2=21.16,P<0.05). Compared to Group A(100%),the survival rate of mice in Group B(0)decreased significantly(P<0.05),and also the pathological damage degrees in the lung,liver and kidney tissues of the mice in Group B increased signifi?cantly after CLP. Compared with the mice in group B,the survival rate of those in Group C(70%)increased significantly(P<0.05),and the pathological damage degrees in the lung,liver and kidney tissues of the mice in Group C decreased significantly after the treatment with AES. The differences among the levels of pro?inflammatory cytokines TNF?α(F=27.11,P<0.05),IL?1β(F=18.75,P<0.05)and IL?6(F=100.93,P<0.05)in the sera of the mice in the three groups were statistically signifi?cant. Compared with the mice in Group A,the levels of the 3 cytokines of those in Group B increased significantly(all P <0.05). However,after the treatment with AES,the levels of the pro?inflammatory cytokines of those in Group C decreased signifi?cantly(all P<0.05). The differences among the levels of immunoregulatory cytokines IL?10(F=10.88,P<0.05)and TGF?β(F=11.37,P<0.05)in the sera of the mice in the three groups were also statistically significant. Compared with the mice in Group B,the levels of IL?10 and TGF?β of those in Group C were higher after treatment with AES(both P<0.05). Conclu?sion T. spiralis AES has a therapeutic potential for alleviating sepsis induced by CLP in mice.
ABSTRACT
A novel electrochemical method based on dehydroabietylamine schiff base( DBS) and multi-walled carbon nano-tubes( MWCNTs) composite modified glassy carbon electrode was presented for determination of Pb2+. The electrochemical behaviors of Pb2+on the modifled electrode were investigated by cyclic voltammetry ( CV) and differential pulse anodic stripping voltammetry ( DPASV ) . The results showed that under the optimized conditions including 0 . 2 mol/L NaAc-HAc used as supporting buffer ( pH 5 . 5 ) , -1 . 1 V of accumulating potential, 250 s of accumulating time, the oxidation peak current was proportional to Pb2+concentration in the range between 1 × 10-8 mol/L and 1 × 10-6 mol/L with the linear regression equation as I(μA)= 6. 6173c(μmol/L)+0. 2597(R=0. 9971) and the detection limit as 5. 0×10-9 mol/L (S/N=3). The proposed sensor was successfully employed to determine Pb2+ in real samples with satisfactory results. In addition, this method showed the advantages of simple operation, short assay time, good accuracy, satisfactory efficiency and good selectivity for determination of low concentration lead in water samples.
ABSTRACT
Objective To investigate the characteristics of growth and angiogenesis of colon cancer xenograft in nude mice with metabolic syndrome induced by high fat diet.Methods Female BALB/C nude mice were fed with high fat diet (45.0％ from fat,HFD group) or common diet (13.8％ from fat,CD group) for 12 weeks (n=15,respectively).Colon cancer cell line SW480 was marked with green fluorescent protein (GFP) and subcutaneous xenograft model was established.The tumor growth was observed by the in vivo imaging system in small animal at the 4th week.By the end of the experiment,serum glucose and lipid level of the two groups were measured,visceral subcutaneous and visceral adipose tissue,liver and xenograft tumor were dissociated and weighted.The differences of proliferating cell nuclear antigen (PCNA) and CD31 expression in the tumors between groups were analyzed.The t-test or x2 test were performed for group comparison.Results Compared with CD group,the body weight,blood serum glucose level,triglyceride and cholesterol level,adipose content of subcutaneous and visceral of the HFD group significantly increased (t=2.91,4.12,4.43,3.92,3.77 and 4.02,all P＜0.05).Averagedaily energy intake of HFD group was significantly higher than that of CD group (t=2.34,P＜0.05).There was no significant difference between the two groups in liver weight (t=1.02,P＞0.05).However,by HE staining lipid vacuoles in the liver tissue was obvious in HFD group.Average bioluminescent index,tumor volume and weight of xenografts of HFD group were remarkably higher than those of CD group (t=8.84,2.48 and 2.86,all P＜0.05).The immunohistochemical staining results indicated that the strong positive rate of PCNA in xenografts of HFD group was 80.00％ and the microvessel density (MVD) was (25.75±0.96)/per high power field,both of which were higher than those of CD group (14.29％ and (13.33±1.53)/per high power field respectively,x2 =12.52,t=13.35,both P＜0.01).Conclusions The colon cancer xenograft in nude mice with metabolic syndrome induced by high fat diet had a high MVD and grew fast.
ABSTRACT
Objective To observe the effects of Toxoplasma lysate antigen(TLA)on growth inhibition and tumor angiogenesis in mice B16 melanoma.Methods Twenty mice bearing B16 melanoma were established by subcutaneous inoculating with B16 cells and were randomly divided into the treatment group and the control group 0.1 ml TLA was administered once every two days for the treatment group starting on the seventh day after inoculation.and an equivalent volume of sterile saline was administered similarly in the control group.The mice were sacrificed on 21 th day after the tumor cells was injected.The anti-tumor effects of TLA were observed by measuring the size and weight of tumor.The microvessel density(MVD)and expression of vascdar endotheilial growth factor(VEGF)were observed by immunohistochemical method.Results TLA Can markedly inhibit the B16 melamoa growth in mice.The volume and weight of the tumors in the experimental group were significantly decreased compared with the control group(P＜0.05),and the turnout inhibitory rate was 49.6%.The MVD of experimental group and control was(44.4000±4.7888)and(31.9000±2.6012)respectively,and with a significant difference between two groups(P＜0.05).The VEGF in the treatment group were significantly lower than those in the control group(P＜0.05).Conclusion TLA can inhibit the growth of the B16 melanoma in mice.suggesting that the anti-tumor effect induced by TLA via a possible antianigionesis mechanism.