Excessive N-acetyl-p-benzoquinone imine(NAPQI)formation is a starting event that triggers oxidative stress and subsequent hepatocyte necrosis in acetaminophen(APAP)overdose caused acute liver failure(ALF).S-glutathionylation is a reversible redox post-translational modification and a prospective mechanism of APAP hepatotoxicity.Glutaredoxin-1(Glrx1),a glutathione-specific thioltransferase,is a primary enzyme to catalyze deglutathionylation.The objective of this study was to explored whether and how Glrx1 is associated with the development of ALF induced by APAP.The Glrx1 knockout mice(Glrx1-/-)and liver-specific overexpression of Glrx1(AAV8-Glrx1)mice were produced and underwent APAP-induced ALF.Pirfenidone(PFD),a potential inducer of Glrx1,was administrated preceding APAP to assess its protective effects.Our results revealed that the hepatic total protein S-glutathionylation(PSSG)increased and the Glrx1 level reduced in mice after APAP toxicity.Glrx1-/- mice were more sensitive to APAP overdose,with higher oxidative stress and more toxic metabolites of APAP.This was attributed to Glrx1 deficiency increasing the total hepatic PSSG and the S-glutathionylation of cytochrome p450 3a 11(Cyp3a11),which likely increased the activity of Cyp3a11.Conversely,AAV8-Glrx1 mice were defended against liver damage caused by APAP overdose by inhibiting the S-glutathionylation and activity of Cyp3a11,which reduced the toxic metabolites of APAP and oxidative stress.PFD precede administration upregulated Glrx1 expression and alleviated APAP-induced ALF by decreasing oxidative stress.We have identified the function of Glrx1 mediated PSSG in liver injury caused by APAP overdose.Increasing Glrx1 expression may be investigated for the medical treatment of APAP-caused hepatic injury.

Objective:To evaluate the effect of 2-aminoethoxydiphenyl-borate (2-APB) on cardiac dysfunction in arsenic poisoning rats by stratified strain technique.Methods:Thirty-two 12-week-old SD rats were randomly divided into control group ( n=8) and arsenic exposure group ( n=24). After 12 weeks of arsenic exposure, arsenic exposure group was divided into three groups: natural recovery group ( n=8), low-dose intervention group (n=8) and high-dose intervention group ( n=8). The rats were treated with 2-APB for 21 days. After the last administration, the routine parameters and the layered strain parameters were measured by ultrasonic instrument. Then the rats were killed and their blood and myocardial samples were obtained. The levels of serum creatine kinase isoenzyme (CK-MB) and lactate dehydrogenase (LDH) were tested, and the morphological changes of cardiomyocytes were observed by HE staining. Results:Left ventricular ejection fraction (LVEF), left ventricular short axis shortening (LVFS), global circumferential strain of subendocardial myocardium (GCS-endo), global circumferential strain of middle myocardium (GCS-mid), global circumferential strain of subepicardial myocardium (GCS-epi) and circumferential strain rate of left ventricular segments (SrC) in the natural recovery group were lower than those in the control group ( P<0.05), while serum CK-MB and LDH were higher than those in the control group ( P<0.05). Some ultrasonic parameters and biochemical indexes in the low and high dose intervention groups were improved in varying degrees compared with the natural recovery group ( P<0.05). The correlation between GCS-endo and CK-MB was the highest ( r=-0.931, P<0.05). Myocardial HE staining showed that the degree of myocardial cell swelling and necrosis were reduced, and erythrocyte exudation was reduced in the low and high dose intervention groups compared with the natural recovery group. Conclusions:The stratified strain technique can be used to evaluate the protective effect of 2-APB on cardiac dysfunction in arsenic poisoning rats, and GCS-endo may be one of its sensitive indexes.

Objective To construct humanized monoclonal antibodies against CD 20 and check their affinity to CD 20 antigen and their anti-tumor activity.Methods Based on the computer model , human IgG1 candidates closest to rituximab in crystal structure were selected in the Protein Data Bank ( PDB) .With the selected human IgG 1 candidates as the frame , we modified and transplanted the complementarity determining region ( CDR) of rituximab .First,the target gene fragments were obtained by overlapping PCR.Then, the sequences of the light chains(L) and the heavy chains(H) were inserted in-to the pcDNA3.3 and pOptiVEC vectors.Next, the constructed clones were transfected into 293F cells through transient transfection.After a large-scale cell culture, the mAb was purified by affinity chromatography rProtein A column.The puri-ty and expression level of the humanized antibodies was tested by sodium dodecyl sulfate ( SDS)-polyacrylamide gelelectro-phoresis(PAGE).The affinity of the humanized antibodies to CD20 was assessed with Fortebio assay.Finally, the anti-tumor activity of the constructed antibodies was detected by checking the tumor growth inhibition of the nude mice transplan-ted with tumor .Results Three humanized monoclonal antibodies against CD 20 were expressed and purified successfully . In reducing SDS-PAGE, the antibodies exhibited two bands of approximately 25 ×103 and 55 ×103 , respectively.The band size of the antibodies matched the expected value.Fortebio assay revealed that the humanized antibodies could bind to CD20 with high affinity (rituximab:6.48 ×10 -9mol/L, L4H7:1.91 ×10 -9mol/L, L5H5:7.35 ×10 -10mol/L,and L5H7:1.91 ×10 -9mol/L).The tumor growth inhibition experiment showed that the anti-tumor activity of L5H7 mAb was better than that of rituximab .Conclusion Three humanized monoclonal antibodies against CD 20 have been successfully construc-ted and expressed.L5H7 mAb possesses high affinity for CD20 and a good ability to kill tumor cells.

Objective To detect the expressions of proliferating cell nuclear antigen (PCNA), C-erbB-2, p53, estrogen receptor (ER) and progesterone receptor (PR) in order to explore the proper margin for breast conservative surgery on Chinese women. Methods We collected 40 resection specimens from breast cancer patients who had received radical surgery. Then we divided each specimen into primary tumor group and hyperplasia and gene expression characteristics of PCNA, c-erbB-2, p53, ER and PR were measured by pathological and immunohistochemical assay in the five groups. Results With the further distance from the primary tumor, the proportions of high-risk disease and positive gene expressions of PCNA, c-erbB-2 and p53 in paracarcinoma tissues gradually decreased (P<0.05). Higher risk and more positive expressions were related to paracarcinoma was no correlation of PCNA, ER and PR expressions adjacent to breast cancer with tissue differentiation and lymph node metastasis (P>0.05). a safe and appropriater region for breast conservative surgery.