Objective To construct a vector containing protein transduction domain (PTD) and cardiotrophin-1 and a vector containing PTD and EGFP, express them in E. coli. and purify them. To detect the distribution of the two fusion proteins in mice. Methods CT-1 and EGFP were cloned to GST-fusion expression vector pGEX-4T3 by PCR and cloning techniques respectively, and then TAT was cloned into the vectors respectively to give pGEX-TAT/CT-1, pGEX-TAT/EGFP. After induced by IPTG the soluble protein GST-TAT/CT-1 and GST-TAT/EGFP was purified by Glutathione Sepharose 4B. The purified fusion proteins were injected into mice through caudal vein and examined in tissue section by immunohistochemical staining. Results CT-1 and EGFP were effectively amplified and the TAT/CT-1 and TAT/EGFP gene sequencing showed the same sequence as scheduled. The fusion proteins was successfully expressed in E. coli. and purified. Conclusion TAT/CT-1 and TAT/EGFP fusion proteins were expressed and purified successfully. The two fusion proteins were all detected positively in mouse brain, spinal cord, heart and liver.

Objective To investigate the value of serum ischemia-modified protein (IMA),hypersensitivity C reaction protein (hs-CRP),myoglobin (MYO)creatine kinase MB (CK-MB)and ultra-sensitivity troponin T (hs-cTnT)in the early diagnosis of patients with acute coronary syndrome(ACS).Methods Serum levels of IMA,hs-CRP,MYO,CK-MB and hs-cTnT were meaused in 94 patients with ACS[including 40 cases of unstable angina pectoris(UAP),20 cases of non-ST segment elevation myocardial in-farction(NSTEMI),non-Q wave myocardial infarction and 34 cases of ST segment elevation myocardial infarction(STEMI),Q wave myocardial infarction]and 99 cases of controls.The efficiency,sensitivity,specificity,negative and positive predictive values in the early diagnosis of ACS were compared among 5 kinds markers by the ROC curve.Results The detection results of serum IMA,hs-CRP,MYO,CK-MB and hs-cTnT had statistically significant differences between the UAP,NSTEMI and STEMI groups with the normal control group (P <0.05),serum IMA,hs-CRP,MYO,CK-MB and hs-cTnT had statistically significant differences between the UAP group with the NSTEMI group and between the UAP group with STEMI group (P <0.05),but the difference between NSTEMI and STEMI group group was not statistically significant (P >0.05).Conclusion In the patients with ACS,the multiple indicators combined detection can achieve the early diagnostic value.

A simple and reliable high performance liquid chromatography (HPLC) method has been developed for the simultaneous quantification of five major bioactive components in ‘Shu-Jin-Zhi-Tong' capsules (SJZTC), for the purposes of quality control of this commonly prescribed traditional Chinese medicine. Under the optimum conditions, excellent separation was achieved, and the assay was fully validated in terms of linearity, precision, repeatability, stability and accuracy. The validated method was applied successfully to the determination of the five compounds in SJZTC samples from different production batches. The HPLC method can be used as a valid analytical method to evaluate the intrinsic quality of SJZTC.

Objective To measure and analyze Varian Clinac 21EX accelerator induced radioactivity,and to provide specific recommendations and ways of radiation protection for radiotherapy technicians.Methods To simulate the working environment of radiotherapy technician,and to detect induced radioactivity variation of Varian Clinac 21EX accelerator that induced by 15 MV X-rays under different conditions of beam field area,dose,time,distance and by high energy electron beam (12,16,20 MeV)at the different time.Results The induced radioactivity level was not influenced by different beam field area,and was increased with increasing dose (r =0.930,P ＜ 0.05),decreased with time increase (r =-0.84,P ＜ 0.05),decreased with distance increase(r =-0.975,P ＜ 0.05).The induced radioactivity attenuation levels of the different doses at the same time are different,and have the common characteristic that the induced radioactivity attenuation rate is faster in the initial times.The induced radioactivity levels of high energy electron beams were significantly lower than those of high-energy X-rays.Conclusions During radiotherapy positioning,it is necessary to take measures to protect against the induced radioactivity when high-energy rays with energy greater than 10 MeV will be used.The radiotherapy technician should take the different time and energy interval steps for the different ray type and energy and dose in order to meet the radiation protection principle of optimization.

AIM: To study the expression and the role of ERK1/2 and JNK1/2 of MAPKs pathways in the development of neural tube defects induced by hyperthermia. METHODS: The animal models of golden hamster were produced by hyperthermia. The expression of ERK1/2 and JNK1/2, and levels of their phosphorylation were measured by Western blotting in control group and hyperthermia group. RESULTS: p-ERK1/2 steadily expressed in each control group, and the expression of p-ERK1/2 significantly decreased, which was different from that in the corresponding control group (P<0.05). The activity of p-JNK1/2 increased in hyperthermia group and the amount of p-JNK1/2 increased as compared to control group. The peak appeared at 16 h after exposed to hyperthermia (P<0.05). CONCLUSION: Hyperthermia, which induces a decrease in p-ERK1/2 expression and increases the expression of p-JNK1/2 of MAPKs pathway, results in the unbalance of cell proliferation and apoptosis, and induces neural tube defects.

Objective To explore the relationship of X-ray exposure parameters and false-node rate during image-guidance treatment with CyberKnife spine tracking.Methods Using spine tracking planning on a chest phantom,several combinations of X-ray exposure parameters were used to locate.The false-node ratio and the surface absorbed dose were investigated and the radiation dose was optimized.Results The false-node ratio and surface absorbed radiation dose decreased when the X-ray exposure parameters increased until they saturated.In the range of ≤5.0％ false-node rate,the surface absorbed radiation dose was 0.11,0.26 mGy,and 0.31-0.46 mGy,when the false-node rate was 2.77％,1.07％and 1.0％,respectively.Conclusions In image-guided treatment of CyberKnife,the radiation dose would be optimized,and the patient's radiation dose would be reduced greatly,which is important to protect the patients.

Objective To investigate the renoprotective effect of transforming growth factor beta activator kinase 1 (TAK1) inhibitor 5Z-7-oxozeaenol (OZ) in diabetic db/db mice and the mechanism.Methods Twenty-four male db/db mice were randomly divided into two groups:db/db mice (db/db,n=12) and db/db mice with 5Z-7-oxozeaenol treatment (db/db+OZ,n=12).Another group of wild type mice (n=12) was held as the control group.OZ 2 mg/kg was administrated by intraperitoneal injection every other day.At week 8 and 12 after 5Z-7-oxozeaenol treatment,blood glucose (BG),body weight (BW),kidney weight (KW) and urinary albumin excretion rate (UAER) were evaluated.Kidney pathological lesions were detected by light and electron microscopy.NF-κB p65,monocyte chemotactic protein-1 (MCP-1) and tumor necrosis factor-ot (TNF-o) were detected by immunohistochemistry.Western blotting was used to detect p-TAK1,TAB1,p-p38MAPK and IL-1β expression,while ICAM-1 and MCP-1 mRNA levels were evaluated by real-time PCR.Results Compared with control group,the levels of BG,BW,KW and UAER were higher (P ＜ 0.01) in db/db mice group,while BW,KW and UAER levels were significantly decreased in db/db + OZ group compared with that in db/db mice group (P ＜ 0.05).In week 8 and 12 db/db mice,glomerular volume and extracellular matrix were increased,while pathological lesions in kidney tissue were positively improved by TAK1 inhibitor.Immunohistochemistry showed that NF-κB p65,MCP-1 and TNF-α expression levels were apparently increased in db/db mice group compared with that in control group (P ＜ 0.05) and were significantly inhibited by TAK1 inhibitor (P ＜ 0.05).Western blotting showed that p-TAK1,TAB1,p-p38MAPK and IL-1β expression levels were higher in db/db mice group than that in control group (P ＜ 0.05) and lower in db/db+ OZ group than that in db/db mice group (P ＜ 0.05).Moreover,real-time PCR showed that the expressions of ICAM-1 and MCP-1 mRNA were higher in db/db mice group than that in control group and lower in db/db+OZ group than that in db/db mice group (P ＜0.05).Conclusions TAK1 Inhibitor can down-regulate MAPK and NF-κB pathway to restrain the reaction of inflammation and alleviate kidney injury in diabetic db/db mice.

Objective To investigate the role of transforming growth factor-β activated kinase-1 (TAK1) signaling pathway in the activation of bone marrow derived macrophages (BMDM) induced by high glucose.Methods Purity of mouse BMDM was detected by flow cytometry.The mice macrophages cultured in vitro were stimulated by high glucose and treated with TAK1 specific inhibitor 5Z-7-oxozeaenol.Cells were divided into normal control group (RPMI 1640),osmolality control group (25 mmol/L mannitol),high glucose group (33 mmol/L D-glucose) and inhibitor group (33 mmol/L D-glucose+300 nmol/L 5Z-7-oxozeaenol).Immunocytochemistry and flow cytometry were used to detect macrophage subtype.The expression of monocyte chemotactic protein-1 (MCP-1) and tumor necrosis Factor-α (TNF-α) mRNA were determined by real time PCR.Expressions of p-TAK1,TAK1 binding protein (TAB1),p-JNK,p-p38 MAPK and NF-κB p65 proteins were analyzed by Western blotting.Results The purity of BMDM was about 99.36％.Compared with normal control group,high glucose group had increased percentage of M1 macrophages,increased expression of MCP-1 and TNF-α mRNA (all P ＜ 0.05).Moreover,p-TAK1,TAB1,p-JNK,p-p38 MAPK and NF-κB p65 proteins expression also increased significantly in high glucose group (all P ＜ 0.05).After treatment with inhibitor 5Z-7-oxozeaenol,the effects induced by high glucose were inhibited (P ＜ 0.05).Conclusions High glucose can induce M1 macrophage activation and expression of inflammatory cytokine of BMDM,which can be inhibited 5Z-7-oxozeaenol through inhibiting TAK1/MAPK and TAK1/NF-κB pathway.

Aim We used bone marrow-derived macro-phages ( BMMs ) , to explore the mechanism of macro-phage activation and the effect of TGF-β activated ki-nase-1 ( TAK1 ) inhibitor 5 Z-7-oxozeaenol on it under AGEs conditions. Methods The BMMs were obtained from C57 mice, and purity of BMMs was detected by flow cytometry. Cell viability was tested after treatment with different concentrations of TAK1 inhibitors. Laser confocal microscopy was used to detect macrophage M1 subtype . Flow cytometry was used to analyse the macro-phage activated by AGEs. TNF-α and MCP-1 mRNA levels were evaluated by qRT-PCR. Western blot was used to detect the expression levels of TAK1 signal pathway protein. Results AGEs stimulation could in-crese the activity of M1 macrophages,and 5Z-7-oxoze-aenol could inhibit the differentiation of BMMs. Com-pared with control group, AGEs increased the expres-sion of MCP-1 and TNF-α mRNA(P<0. 01). p-TAK1, TAB1,p-JNK,p-p38MAPK and NF-κBp65 proteins ex-pression also increased significantly ( P <0. 05 ) . After treatment with inhibitor, transcription levels of MCP-1 and TNF-α decreased significantly ( P < 0. 05 , P <0. 01 ) . 5 Z-7-oxozeaenol treatment downregulated the expression of p-TAK1,TAB1,p-JNK,p-p38MAPK and
NF-κBp65 proteins ( P <0. 05 ) . Conclusions AGEs can induce BMMs to M1 phenotypic polarization. 5Z-7-oxozeaenol reduces the expression of inflammatory cyto-kine via inhibiting TAK1/MAPKs, MAPKs/NF-κB pathways.

Objective: To determine the valuable hemolytic characteristics in differential diagnosis of paroxysmal nocturnal hemoglobinuria (PNH), autoimmune hemolytic anemia (AIHA) and hereditary spherocytosis (HS). Method: The clinical and hemolytic characteristics of 108 PNH patients, 127 AIHA patients and 172 HS patients diagnosed from January 1998 to April 2017 were compared. Results: ①Reticulocyte percentage (Ret%) of PNH patients [6.70% (0.14%-22.82%)] was significantly lower than that of AIHA [14.00%(0.10%-55.95%), P<0.001] and HS patients [11.83%(0.60%-57.39%), P<0.001]. The Ret% in PNH patients were significantly lower than those in AIHA and HS patients at the same levels of anemia, except for in mild anemia between PNH and AIHA patients. However, when comparing the Ret% between AIHA and HS patients, there was significant difference only in mild anemia [7.63%(1.87%-29.20%)% vs 11.20%(3.31%-22.44%), z=-2.165, P=0.030]. ②The level of TBIL in HS patients was significantly higher than that in AIHA and PNH patients [79.3 (11.2-244.0) μmol/L vs 57.6 (7.6-265.0) μmol/L, z=5.469, P<0.001; 79.3(11.2-244.0) μmol/L vs 26.2(4.6-217.7) μmol/L, z=-2.165, P<0.001], and the proportion of HS patients with TBIL more than 4 times the upper limit of normal (ULN) (64.1%) was significantly higher than that of AIHA (37.7%, χ²=19.896, P<0.001) and PNH patients (4.6%, P<0.001). ③The LDH level of PNH patients was significantly higher than that of AIHA and HS [1 500 (216-5 144) U/L vs 487 (29-3 516) U/L, z=-9.556, P<0.001; 1 500 (216-5 144) U/L vs 252 (132-663) U/L, z=-11.518, P<0.001], and the proportion of PNH patients with LDH more than 1 000 U/L (79.1%) was significantly higher than that of AIHA patients (13.0%, χ²=93.748, P<0.001) and HS patients (0, P<0.001). ④Splenomegaly occurred in 43.5% of PNH patients, including 16.0% with severe splenomegaly. In contrast, the occurrence of splenomegaly was 98.6% in AIHA patients and 100.0% in HS patients (P<0.001), and 63.0% of AIHA patients (P<0.001) and 90.4% of HS patients (P<0.001) were with severe splenomegaly. ⑤The prevalence of cholelithiasis in HS patients was up to 43.1%, significantly higher than that in AIHA patients (10.5%, P<0.001) and PNH patients (2.9%, P<0.001). Conclusion The comprehensive assessment of the five hemolytic characteristics is simplified, practical and efficient, with great clinical significance, providing specific indicators for differential diagnosis and efficient approach for making further work-up.