Objective:To explore the Wnt-β-catenin signal molecule expression level in acute kidney injury rats renal tissue. Methods:A total of 48 rats accorded to the random number table method were divided into normal control group ( Control group) ,sham operation group ( Sham group) and ischemia reperfusion group ( IRI group) ,each group with 16 rats,Control group was given normal fed,IRI group were established ischemia reperfusion injury rats model,Sham group opened dorsal skin and then wound was closed. 4 rats were sacrificed respectively at 1,3,5,7 d after surgery,the pathological changes of kidney tissue were observed in the three groups,and the serum creatinine,urea level was detected in the three groups. Wnt4,β-catenin,Lrp6 protein expression was detected in three groups of rats by Western blot,Wnt4,β-catenin,Lrp6 mRNA in renal tissues were detected by Real-time quantitative RT-PCR. Results:There was no significant difference in renal injury score and serum creatinine between Control group and Sham group (P>0. 05),IRI group kidney injury score and serum creatinine,urea first increased and then gradually decreased,which was the highest in 3 d after surgery,IRI group renal injury score and serum creatinine,urea were significantly higher than those in Control group and Sham group 1,3,5,7 d after surgery (P<0. 05). There was no significant difference in Wnt4,β-catenin,Lrp6 protein between Control group and Sham group (P>0. 05),IRI group Wnt4,β-catenin,Lrp6 protein gradually increased,which reached to peak at 5 d after surgery,and then gradually decreased,IRI group Wnt4,β-catenin,Lrp6 protein were significantly higher than those in Control group and Sham group 1,3,5,7 d after surgery(P<0. 05). There was no significant difference in Wnt4,β-catenin,Lrp6 mRNA between Control group and Sham group (P>0. 05),IRI group Wnt4,β-catenin,Lrp6 mRNA gradually increased,which reached to peak at 3 d after surgery,and then gradually decreased,IRI group Wnt4,β-catenin,Lrp6 mRNA were significantly higher than those in Control group and Sham group 1,3,5,7 d after surgery (P<0. 05). Conclusion: The expression of Wnt-β-catenin signal molecule are significantly increase in acute kidney injury,and the activation of Wnt-β-catenin signal pathway is involved in the repair process of renal tissue.

In order to study the role played by infection in the occurrence and mechanism of negative nitrogen balance of the skeletal muscles in the early stage of scalding,37% TBSA full thickness scalding was inflicted on the back and one hind leg of rats and the protein catabolic rate in the soleus muscle of both the scalded and healthy legs was determined and the effects of endotoxin on the catabolic rate were observed.It was found that the protein catabolic rate was markedly accelerated in the scalded soleus 3 days after iujusy;and showed no significant changes in the healthy one,in which the protein catabolic rate was near or similar to that of the normal control.After 2 mg/kg of endotoxin was administered intraperitoneally.the findings were as follows:(1)Endotoxin could also accelerate the protein catabolic rate in the skeletal muscles;(2)The protein catabolic rate in the scalded soleus was not further increased and no summation effect of scalding and endotoxin on protein catabolism occurred.

Objective To explore the pathological features and the causative particles of severe acute respiratory syndrome (SARS) for providing evidences of SARS prevention and clinical treatment. Methods A dead case of SARS of China was studied by light microscopy, electron microscopy, histochemical and immunohistochemical stain. Results The major pathological changes of lung in the SARS case were acute pulmonary interstitial exudative and leakage inflammation, with predominant lymphocyte infiltration. The hyaloid membranes were formed in 20%～30% pulmonary alveoli. The diffuse pulmonary epithelial injury was observed, and virus like inclusions were found in about 30% of total alveolar epithelia by histochemical stain, but chlamydia like inclusions were found occasionally. Meanwhile, the extra pulmonary organs, such as lymph nodes and spleen, showed extensive haemorrhagic necrosis inflammation, accompanied macrophage/histocyte reactive proliferation with erythrocytophage. The double adrenal glands also presented focal haemorrhagic necrosis inflammation. Under the electron microscopy observation, virus like particles with 100 ～150 nm diameter and halo or garland envelopes were found in more than 30 % alveolar epithelial cells, endothelial cells in lung tissues, and also in a part of cardiomyocytes, lymphocytes and macrophages in lymph nodes. The virus like particles were mainly located in cytoplasm and dilated reticular endoplasm. In contrast, chlamydia like particles were commonly visualized in multiple extra lung organs such as liver, but very few in the lung. Immunohistochemistry showed the positive reactions in the lung tissues with the serum IgG and/or IgM from the dead case himself and other SARS convalescent stage cases from Guangdong province of China. Conclusion In the severe SARS case, predominant acute interstitial exudative and leakage inflammation, often with the formation of hyaloid membranes in pulmonaryalveoli, and the haemorrhagic necrosis inflammation of immune organs might be pathological features of SARS. According to the structures, diameter and location of the virus like particles found in this case, combined with the pathological changes, we should consider that those virus like particles might be a new kind of coronavirus, and this kind of virus might be the main causative agent of SARS. However, the chlamydia like particles frequently observed in extra lung organs also suggested the potential new kind of coronavirus might be coexist and synergicallly cause SARS. Our findings in this study provide several evidences for SARS clinical therapy such as application of corticosteroid and enhancement of immune ability and combination of anti virus and anti chlamydium drugs.

Objective To study the pathological changes in organs remote from the lung in SARS patient. Methods The pathological changes in extra lung organs and potential coronavirus infection were studied by using light and electron microscopic examinations as well as special virus inclusion stains in the tissues obtained from an autopsy of a patient who died of SARS. Results Besides the lesions in the lung, pathological changes were found also in the central nervous system (CNS), including the cerebrum, cerebellum, thalamus, pons, and medulla oblongata, such as widening of the Virchow Robin′s space, infiltration of a few lymphocytes and macrophages in the parenchyma, vasodilatation and congestion. However, no significant neuron degeneration or necrosis was identified. Vasodilatation in the lamina propria of mucosa and submucosa of the digestive tract with some lymphocytes infiltration, and epithelial nuclear vacuolation, and occasional apoptosis were observed in the mucosal epithelial and glandular cells, as well as focal hemorrhage in segments of the small intestine. Mesenchymal edema and infiltration of a few lymphocytes in the pancreas were noted. Very mild lymphocyte infiltration, but no viral inclusions, was found in the convoluted seminiferous tubules of the testis. The patient who died of SARS was proved to have arteriosclerosis of the coronary arteries, and coronavirul particles were identified in the blood vessels under electron microscopic examination, however no coronavirul particles were found in the brain or the testis of the patient. Conclusion There were mild hypoxic changes in the tissue of CNS in the patient with severe SARS without invasion of the virus. It was confirmed that there were coronavirul particles in the blood of the patient at the acute stage of SARS. Since the patient who succumbed to the disease had a history of coronary arteriosclerosis, it was inferred that cardiovascular disease might be a contributory factor of mortality in this patient with severe SARS.

Objective To explore the target cells of SARS coronavirus infection in vivo and to provide the evidence of multi organ injuries produced by SARS coronavirus infection. Methods Three biotin labeling oligonucleotide probes were synthesized according to the published gene sequence of SARS coronavirus. The location, distributtion and quantity of SARS coronavirus in 2 autopsy cases of SARS were studied by in situ hybridization and electron microscopic examination. Results SARS coronavirus particles were identified in multiple organs. In lungs, SARS coronaviruses were located predominantly in the cytoplasm of bronchiolar and alveolar epithelial cells, in a part of macrophages and endothelial cells as well as a few infiltrated lymphocytes. In situ hybridization showed that in target cells SARS coronavirus distribution presented a cytoplasmic or inclusive pattern, and the mean number of positive cells in the pulmonary tissue was 80?25 per 200? field. Electron microscopic examination showed that the coronaviral particles were 100～150 nm in diameter, with low density electron cores with halo or garland envelopes. About 15% of renal tubular epithelial cells harbored SARS coronavirus, and a few parenchymal cells and sinusoid capillary endothelial cells of adrenal glands were hybridization positive. In the gastro intestinal tract, SARS coronaviruses were seen in the cytoplasm of mucosal and crypt epithelial cells, mostly in 2/3 of superficial mocosa. Under both electron microscopy and in situ hybridization observation, SARS coronaviruses were found focally distributed in some cardiomyocytes. The SARS coronavirus positive particles were also noted in macrophages/histocytes, sinusoid endothelial cells, as well as a few lymphocytes in thoracic and celiac lymph nodes. In addition, coronavirus particles were also seen in a few testicular epithelial cells and Leydig's cells. Conclusion SARS coronavirus could attack multiple target cells, implicating that SARS might cause multi organ damages, with lungs as the predominant organ of injury.

Objective To study whether transient overexpression of tumor suppressor gene PTEN could lead to growth suppression and up-regulate the sensitivity to doxorubicin of human breast cancer MCF-7 cells. Methods The eukaryotic expression plasmid pEGFP-C 1-PTEN containing whole cDNA of PTEN was constructed and transfected into MCF-7 cells by Lipofectamine 2000 in vitro. Growth inhibition of the cells was observed by phase contrast microscope and flow cytometry. The clonogenic cell survival ability was studied by clony forming assay. MCF-7 cells′ chemosensitivity to adriamycin was studied with MTT assay. Results PTEN overexpression led to morphological changes characteristic of apoptosis of MCF-7 cells. PTEN overexpression also resulted in a significant increase in G 0/G 1 cell population (14.79%) and apoptosis (10.60%) detected by flow cytometry. The clonogenic survival rate of cells transfected with PTEN was significantly decreased after doxorubicin treatment compared with control. The transfected cells were more sensitive to doxorubicin compared with the control cells ( ? 2=8.59 , P

Lectin-likeoxidizedLDLreceptor-1(LOX-1)isoneofthemajorreceptorsoftheoxidized low-density lipoproteins (oxLDL). Recent research suggests that LOX-1 can not only bind, neutralize and degrade oxLDL, but also bind some non-oxidized lipoproteins and apoptotic blood cel s. LOX-1 is involved in the process of atherosclerosis through a variety of w ays, such as activating platelet, inducing inflammatory reaction, promoting the proliferation and migration of vascular smooth muscle cels. This article reviews the related research progress from the relationship betw een the structure and function of the LOX-1 and its lipid metabolism and the pathogenesis of atherosclerosis.

Objective To explore the applicability of the childhood autism spectrum test (CAST) of children in Mainland China according to autism screening,in order to provide the effective reference for the identification and diagnosis of autism spectrum disorders of children.Methods Two hundreds and fifty-two typical developing children,and 61 autistic children who were confirmed by psychological clinic of Child Healthcare Clinic,Shenzhen Luohu District Maternal and Child Health Care Hospital who were conformed with diagnosis standards of Diagnostic and Statistical Manual of Mental Disorders Fifth Edition(DSM-5) by using CAST Chinese version were assessed,and the reliability and validity analysis were made on the CAST scale.The receiver operating characteristic curve was drew to determine the optimal threshold value,and the sensitivity,specificity and positive predictive value were calculated.Results CAST Chinese version had good reliability,Cronbach's α was 0.880.Split-half reliability coefficient r was 0.821,and the whole scale calibration reliability coefficient was 0.902.There was statistically significant difference between items and the total score(P＜0.05).The consistency between the CAST ≥4 years old children and diagnosis standards of DSM-5 reached 0.884(P=0.000).The consistency 3-＜4 years old children and diagnosis standards of DSM-5 reached 0.867 (P=0.000).Among the screening of the children who were ≥4 years old,the cut off was 15 scores that the sensitivity and specificity were best.With the children of 3-＜4 years old,a designated cut-point should be 17 scores.Conclusions There are good reliability and validity of CAST,which can be used to mainland China,as well as to screen autistic children more than 3 years old.

Aim To study the effects of polyamine analogue CPENSpm on the human lung cancer line A549 in cell proliferation and apoptosis.Methods MTS was used to assay the cell proliferation,chemical analysis methods were used to determine the activities of enzymes in the polyamine metabolism,HPLC was performed to assay the intracellular concentration of polyamines,Sub-G1 and DNA fragmentation assays were used to determine the cell apoptosis.Results Treating A549 lung cancer cells by CPENSpm resulted in:①cell-growth inhibition and cell apoptosis;②inhibition of ODC(key enzyme in polyamine synthetic pathway)and activation of SSAT and SMO(key enzymes in polyamine catabolism);③great decrease of intracellular polyamine concentrations.MDL72527,the SMO inhibitor,can antagonize the effect of CPENSpm on inhibiting the proliferation of A549 cells.Conclusion CPENSpm inhibits proliferation and induces apoptosis of human A549 lung cancer cell line by interfering the polyamine metabolism,depleting intracellular polyamine contents that are need by quick-growth of cancer cells and inducing production of H2O2.

The Caco-2 cell model was widely applied in the research of absorption,metabolism and toxicity of drugs, especially in the aspect of anticancer,inorganic and traditional Chinese medicine,It has become an important tool of the study on medicine.