Objective To investigate the efficacy of photodynamic therapy(PDT) for tumor diagnosis.Methods A total of 1 400 patients with the cervix of uterus disease,56 patients with gullet disease,133 patients with stomach diseases,37 patients with urinary bladder tumor and 14 patients with brain tumor underwent diagnosis with PDT,combination of He-cd laser,Ar+ laser,and KTP laser.Results All of the patients could be explicitly diagnosed by this method,which indicated that PDT helped greatly in early tumor diagnosis. Conclusion PDT can be an effective diagnostic method for tumors of early stage,and should be widely applied clinically.

The purpose of this study was to investigate and analyze the high frequency electrocardiogram (HFECG) for middle-aged and elderly people.138 subjects were chosen in our study.The mean age was 60.38 years.Among them,72.1 per cent were between 50 and 69 years.The results showed that (1) the differences were not significant between male and female groups in the mean number of high frequency notches (HFN).(2) For an increase in high frequency notches in the leads V1 and V2,we should consider the possibility of pathological changes of the anterior medial wall of heart,which was supplied by the dessending rami of the right anterior coronary artery,on the one hand,and it might remind us to pay great attention to posteromedian wall of heart,on the other hand.(3) Looking at all age groups in this study,we preliminarily found that the age peak of ischemic heart disease was between 50 and 70 years,according to the number of high frequency notches in the leads of HFECG.(4) The sensitivity of HFECG for diagnosing ischemic heart disease is 40 per cent higher than that of the conventional ECG.(5) In addition,nine leads were used in this examination,which not only preserved sensitivity of six leads but also could provide a localizing value for the diagnosis of heart disease.

Objective To explore the effect of azelastine hydrochloride nasal spray combined with desloratadine to inflammatory factors, cell function and IgE of patients with allergic rhinitis.Methods 92 cases of allergic rhinitis patients treated in the first affiliated hospital of wenzhou medical college hospital from June 2014 to December 2015 were divided into experimental group(n=46) and control group(n=46) according to the random number table method.The control group was given oral loratadine tablets, one piece per time, one time per day, while the experimental group was given azelastine hydrochloride nasal spray on the basis of the control group,each nostril one spray, one time in the morning and night.The clinical efficacy of two groups of patients would be observed after 4 weeks,ELISA would be used to detect serum levels of IFN-γ, IL-4, IL-8 and IgE level, and IFN-γ/IL-4 was the value of Thl/Th2,flow cytometry instrument was used to the determination of T cell subgroup CD4 +,CD8 + cells.Results 4 weeks after treatment,stuffy nose, nasal itching, runny nose, sneezing and nasal cavity change points are lower than before the treatment in both the two groups.Experimental group obviously lower than the control group, the difference was statistically significant ( P <0.05 ).The total effective rate of treatment group is higher than the control group,the difference was statistically significant(χ2 =4.389,P=0.036).The serum level of IFN-γis higher than before treatment in both the two groups.IL-4, IL-8 inflammatory factor levels were lower than before treatment,the experimental group was better than control group,the difference was statistically significant(P<0.05).CD4 +,CD8 +of T cells and Thl/Th2 values are higher than before the treatment in both the two groups,the experimental group was higher than control group, the difference was statistically significant ( P<0.05 ).Serum IgE levels were lower than before the treatment in both the two groups,the experimental group was lower than control group,the difference was statistically significant (P<0.05).Conclusion The therapy of azelastine hydrochloride nasal spray combined with desloratadine can improve the clinical effect of the treatment of allergic rhinitis,reduce inflammation,strengthen the body's immune function, improve thelevel of serum IgE.

OBJECTIVE: To study the mTOR expression of cancer stem cells(CSCs) in nasopharyngeal carcinoma and preliminarily explore the mechanism of inhibiting its proliferation with rapamycin. METHOD: Nasopharyngeal carcinoma spherical cells were gathered by using serum-free suspension culture method, CCK8 assay was used to detect cell proliferation, Western blot assay was used to detect the expression of CD44, OCT4, SOX2 and mTOR signaling. The spherical cells and CNE2 were treated with rapamycin in concentrations of 0, 0.1, 1.0, 10.0, 100.0, 1000.0 nmol/L, CCK8 assay was used to detect cell inhibition ratio, Western blot assay was used to detect the expression of mTOR signaling of nasopharyngeal carcinoma spherical cells. RESULT: Compared with CNE2, the spherical cells exhibited a high proliferation rate in RPMI 1640 medium supplemented with fetal bovine serum, and overexpressed in OCT4, SOX2 (P < 0.05), but not that of CD44 (P > 0.05). Although the expression levels of mTOR, P70S6, 4EBP1 were not significantly different between the two kinds of cells (P > 0.05) the proteins of phosphorylation activation form of them (P-mTOR, P-P70S6, P-4EBP1) were highly expressed in spherical cells (P < 0.05). The spherical cells and CNE2 were treated with rapamycin in different concentrations, the concentrations for 50% of maximal effect of spherical cells and CNE2 were 2.59 nmol/L and 78.12 nmol/L respectively, rapamycin inhibited the spherical cells more strongly compared with CNEZ. The expression levels of P-mTOR, P-70S6, P-4EBP1 in spherical cells were gradually decreased with increasing of the concentrations of rapamycin, but the difference of the expression levels of mTOR, P70S6, 4EBP1 were not significant. CONCLUSION The proteins of mTOR signaling pathway of CSCs in nasopharyngeal carcinoma are overexpressed, and rapamycin can effectively inhibit cell proliferation of CSCs in nasopharyngeal carcinoma by blocking mTOR signaling pathway.
Adaptor Proteins, Signal Transducing ; metabolism ; Carcinoma ; Cell Proliferation ; Humans ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplastic Stem Cells ; drug effects ; metabolism ; Phosphoproteins ; metabolism ; Phosphorylation ; Signal Transduction ; Sirolimus ; pharmacology ; TOR Serine-Threonine Kinases ; metabolism ; Tumor Cells, Cultured

Objective To investigate whether heparin has a beneficial effect on lipopolysaccharide(LPS)-induced acute lung injury(ALI)in rats,and to explore the possible underlying mechanisms. Methods Thirty-two adult Sprague-Dawley(SD)rats were randomly assigned into the control,heparin control,model,and heparin treatment groups,with 8 in each group. ALI rat model was reproduced by intratracheal instillation of LPS at a dose of 1 mg/kg. The rats in the control and heparin control groups received an equal volume of normal saline at the same times. The rats in the heparin control and heparin treatment groups were intravenously received 50 U/kg heparin every 1 hour after the induction of ALI. Animals were sacrificed 24 hours after LPS challenge. Bronchoalveolar lavage fluid(BALF) and lung tissue samples were collected. Histopathological evaluation,lung wet/dry(W/D)ratio,malondialdehyde (MDA),nitric oxide(NO)and myeloperoxidase(MPO)were analyzed. Enzyme-linked immunosorbent assay(ELISA) was used to measure the concentration of inflammatory factor in BALF. Expression of inducible nitric oxide synthase (iNOS)mRNA in the lung of rats was measured by reverse transcription-polymerase chain reaction(RT-PCR). Western Blot was used to determine the expression of transforming growth factor-β1(TGF-β1)and phosphorylation of Smad in the lung tissues. The expression of iNOS in lung was determined by immunohistochemistry. Results In the control and heparin control groups,lung tissue showed a normal structure and clear pulmonary alveoli under a light microscope. In the model group,ALI characters such as extensive thickening of the alveolar wall,significant infiltration of inflammatory cells,demolished structure of pulmonary alveoli,and hemorrhage were found. In the heparin treatment group,heparin treatment markedly alleviated LPS-induced these pathological changes in lung. Compared with control and heparin control groups,lung W/D ratio,lung MDA,NO and MPO levels,and tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6)in BALF in the model group were increased significantly. Compared with the model group, lung W/D ratio,lung MDA,NO and MPO levels,and TNF-αand IL-6 in BALF in the heparin treatment group were significantly decreased〔W/D ratio:7.54±0.17 vs. 10.69±0.15,MDA(mmol/mg):2.01±0.30 vs. 2.51±0.25,NO (μmol/L):3.07±0.21 vs. 3.89±0.14,MPO(U/g):1.94±0.09 vs. 2.74±0.20,TNF-α(μg/L):201.80±0.27 vs. 297.53±0.34,IL-6(μg/L):38.41±0.25 vs. 46.31±0.31,all P<0.05〕. RT-PCR showed that the expression of iNOS mRNA in the heparin treatment group was significantly lower than that in the model group(2-ΔΔCt:3.04±0.18 vs. 4.37±0.15,P<0.05). Western Blot showed that compared with control group,the protein expressions of iNOS and TGF-β1,and phosphorylation of Smad2 and Smad3 were significantly increased,and the heparin could inhibit the protein expressions compared with model group. Immunohistochemistry showed that positive expressions of iNOS in alveolar epithelial cell and capillary endothelial cell in the heparin treatment group were significantly lower than those in the model group. Conclusion Heparin significantly ameliorated the lung injury induced by LPS in rats via the inhibition of nitric oxide synthase expression and the TGF-β/Smad pathway.

To develop the stable transformants of the silkworm (Bombyx mori) BmN cells that could continuously express the exogenous gene based on a non-transposon vector, an expression cassette containing human granucyto-macrophage colony-stimulating factor (hGM-CSF) gene driven by ie-1 promoter from B. mori nucleopolyhedrovirus was inserted into pIZT-V5-His to form a recombinant vector pIZT-IE-hGM-CSF, followed by transfecting the constructant into BmN cells, the stable ie-hGM-CSF cell lines were obtained after being selected with Zeocin. PCR result using the genomic DNA of the transformed BmN cells as template illustrated a specific fragment of ie-hGM-CSF, and Western blotting analysis using an antibody against hGM-CSF demonstrated a specific band with a molecular weight of 22 kDa in the transformed cells, meanwhile, the expression level of hGM-CSF determined by ELISA was about 2 814.7 pg in 10(6) transformed BmN cells.
Animals ; Animals, Genetically Modified ; Bombyx ; cytology ; genetics ; metabolism ; Cell Line ; Fibroins ; genetics ; Genetic Vectors ; genetics ; Granulocyte-Macrophage Colony-Stimulating Factor ; biosynthesis ; genetics ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transformation, Genetic

AIM:The objective of this study was to observe the effects of AP-1 element on endothelin-1(EDN1)gene transcription in homocysteine(Hcy)-induced human umbilical vein endothelial cells(HUVECs).METHODS:The redox level in Hcy-induced HUVECs was determined by using the fluorescent product DCF.The influences of Hcy on expressions of EDN1 mRNA and protein of c-jun/AP-1 in HUVECs were measured by the methods of RT-PCR and Western blotting,respectively.The EDN1 secretion level of HUVECs induced by Hcy was determined by enzymatic immunoassay.The transient transfection assay was performed and luciferase activity of transcriptional factor AP-1 reporter gene induced by Hcy was detected.RESULTS:The Hcy significantly increased EDN1 secretion,EDN1 mRNA expression and oxidative stress in HUVECs.Hcy remarkably induced the expression of pGL3-EDN1-AP-1(115/+135)luciferase reporter gene plasmid.However,basic expression of mutation of pGL3-EDN1-AP-1(-115/+135)luciferase reporter gene plasmid was downregulated markedly in HUVECs induced by Hcy.CONCLUSION:Redox-active and release of ROS are changed by Hcy.Activated AP-1 element plays an important role in EDN1gene transcription in HUVECs induced by Hcy.

Objective To retrospectively analysis the clinical effect of temporary hemiepiphysiodesis for treatment of genu varus and valgus with pathologic physis in children.Methods All of 31 children (52 knees) were included in the study from January 2008 to December 2014,20 boys and 11 girls,the age at the time of surgery from 2 year 4 month to 13 year 4 month,mean 6 year 5 month.12 varus and 19 valgus,10 unilateral and 21 bilateral,41 femurs and 44 tibias.The Mechanichal Axis Deviation (MAD) was evaluated and the mechanical lateral distal femur angle (mLDFA) and medial proximal tibia angle (mMPTA) pre-operation and post-surgery was measured.Judging the efficacy with zone system for assessing mechanical axis based on Stevens,we think the results was satisfactory if the mechanical axis falls in zone ± 1 when removed the eightplate and other was unsatisfactory.We made statistical analysis of children who were corrected satisfactory and observed the change of the width and appearance of the physis.We also made the univariate analysis and Logistical multivariate regression analysis about sex,unilateral or bilateral,surgery age,femur or tibia,varus or valgus and severity of deformity to judge the influencing factors between satisfactory and unsatisfactory.We recorded the knee function and deformity recurrence at last follow up.Results The follow-up period was 2 year 6 month to 9 year 5 month,mean 4 year 1 month,44 knees (84.6％) were corrected satisfactory.The mean value of MAD at pre-surgery and at the time when the plate was removed were (-33.3t7.2) mm and (2.1 ±4.3) mm,mean mLDFA were 102.9°±4.9° and 85.3°±3.8°,mean mMPTA were 81.2°±3.4° and 90.5° ±4.4° in genu varus (21 knees);The meam value of MAD at pre-surgery and at the time when the plate was removed were 29.3±6.8 mm and-4.1±6.5 mm,and the mean mLDFA were 79.5o±5.7° and 88.1°±3.5°,mean mMPTA was 97.0°±4.3° and 87.1°±5.2° in genu valgus (23 knees),which were significant difference.There were 28 knees with physis width asymmetry in the 44 knees pre-operation and improved obviously after-surgery and the appearance became normally.8 knees corrected unsatisfactory included 2 varus and 6 valgus.The severity of deformity was a risk factor that affects efficacy with univariate analysis(t=5.124,P=0.000).Regarding the range of the age,we also did the logistic multivariate regression analysis with results showing that surgery age (OR=1.032) and se verity of deformity (OR=1.335) were related to the outcome.There were 8 knees in the 44 knees which had deformity recurrence and the rate was 19.5％ (8/41,3 knees excluded due to doing the limb lengthening surgery when removing the plate) at last follow up,and the knees function were normal.Conclusion Temporary hemiepiphysiodesis is a minimally invasive and effective method for treatment of genu varus or valgus with pathologic physis in children and could improve the appearance of the physis.The age at surgery and severity of deformity are the risk factors that affect efficacy.The pathological changes persisted could cause complication and deformity recurrence which need follow-up after removal of the plate.

Background/Aims: Ubiquitination is widely involved in the progression of hepatocellular carcinoma (HCC) by regulating various cellular processes. However, systematic strategies for screening core ubiquitin-related genes, clarifying their functions and mechanisms, and ultimately developing potential therapeutics for patients with HCC are still lacking. Methods: Cox and LASSO regression analyses were performed to construct a ubiquitin-related gene prediction model for HCC. Loss- and gain-of-function studies, transcriptomic and metabolomics analysis were used to explore the function and mechanism of UBE2S on HCC cell glycolysis and growth. Results: Based on 1,423 ubiquitin-related genes, a four-gene signature was successfully constructed to evaluate the prognosis of patients with HCC. UBE2S was identified in this signature with the potential to predict the survival of patients with HCC. E2F2 transcriptionally upregulated UBE2S expression by directly binding to its promoter. UBE2S positively regulated glycolysis in a HIF-1α-dependent manner, thus promoting the proliferation of HCC cells. Mechanistically, UBE2S enhanced K11-linkage polyubiquitination at lysine residues 171 and 196 of VHL independent of E3 ligase, thereby indirectly stabilizing HIF-1α protein levels by mediating the degradation of VHL by the proteasome. In particular, the combination of cephalomannine, a small molecule compound that inhibits the expression of UBE2S, and PX-478, an inhibitor of HIF-1α, significantly improved the anti-tumor efficacy. Conclusions UBE2S is identified as a key biomarker in HCC among the thousands of ubiquitin-related genes and promotes glycolysis by E3 enzyme-independent ubiquitination, thus serving as a therapeutic target for the treatment of HCC.

Background/Aims: Ubiquitination is widely involved in the progression of hepatocellular carcinoma (HCC) by regulating various cellular processes. However, systematic strategies for screening core ubiquitin-related genes, clarifying their functions and mechanisms, and ultimately developing potential therapeutics for patients with HCC are still lacking. Methods: Cox and LASSO regression analyses were performed to construct a ubiquitin-related gene prediction model for HCC. Loss- and gain-of-function studies, transcriptomic and metabolomics analysis were used to explore the function and mechanism of UBE2S on HCC cell glycolysis and growth. Results: Based on 1,423 ubiquitin-related genes, a four-gene signature was successfully constructed to evaluate the prognosis of patients with HCC. UBE2S was identified in this signature with the potential to predict the survival of patients with HCC. E2F2 transcriptionally upregulated UBE2S expression by directly binding to its promoter. UBE2S positively regulated glycolysis in a HIF-1α-dependent manner, thus promoting the proliferation of HCC cells. Mechanistically, UBE2S enhanced K11-linkage polyubiquitination at lysine residues 171 and 196 of VHL independent of E3 ligase, thereby indirectly stabilizing HIF-1α protein levels by mediating the degradation of VHL by the proteasome. In particular, the combination of cephalomannine, a small molecule compound that inhibits the expression of UBE2S, and PX-478, an inhibitor of HIF-1α, significantly improved the anti-tumor efficacy. Conclusions UBE2S is identified as a key biomarker in HCC among the thousands of ubiquitin-related genes and promotes glycolysis by E3 enzyme-independent ubiquitination, thus serving as a therapeutic target for the treatment of HCC.