Objective:To investigate the effect of 12(S)-HETE on the p27~(kip1) expression in mesangial cells and glomeruli.Methods:Mesangial cells were exposed to 12(S)-HETE.12(S)-HETE was infused to rats by osmotic mini-pump.Total protein content measurement for cell hypertrophy,RT-PCR for mRNA expression and Western blot for protein expression were performed respectively.Results:12(S)-HETE stimulation induced mesangial cell hypertrophy and p27~(kip1) protein expression,but not p27~(kip1) mRNA expression.Furthermore,p27~(kip1) mRNA and protein expression in the glomeruli were significantly increased by 12(S)-HETE stimulation using osmotic mini-pump.Conclusion:12(S)-HETE plays an important role in the pathogenesis of glomerular cell hypertrophy and senescence through upregulation of p27~(kip1) expression.

Objective To investigate the feasibility of using anti-sense RNA against classⅡmajor histocompatibility complex(MHCⅡ)transactivator(CⅡTA),which might regulate MHCⅡexpression,to suppress the relative immune response.Methods Stable transfectants of dermal fibroblasts with pDarⅡ(pDarⅡ-D)were tested for the expression of classic MHCⅡ(HLA-DR,-DP,-DQ)antigens induced with recombinant human interferon-gamma(IFN-?).mRNA abundance of CⅡTA,and classic MHCⅡwas mea-sured by RT-PCR.IL-2mRNA expressed in T cells,stimulated by transfected dermal fibroblasts,was de-termined by mixed lymphocyte reaction.Results When induced with IFN-?,the expression of HLA-DR and-DP antigens on pDarⅡ-D was reduced by95.63%and87.89%,respectively.Meanwhile,the mRNA contents of CⅡTA and classic MHCⅡwere decreased significantly(P

Objective To investigate the clinical value of myocardial enzymes and umbilical artery blood gas analysis in the diagnosis and treatment of neonatal asphyxia (HIE).Methods Eighty-seven neonates with HIE and 94 healthy neonates(control group) were selected and compared.The myocardial enzymes [aspartate aminotransferase (AST),creatine phosphokinase(CK),creatine phosphokinase isoenzyme (CK-MB),hydroxybutyrate dehydrogenase (HB-DH),lactate dehydrogenase (LDH)] and umbilical artery blood gas analysis [pH,alkali residue (BE),arterial oxygen pressure(PaO2),arterial carbon dioxide partial pressure (PaCO2)] of the two groups were detected and compared.Results The levels of AST,CK,CK-MB,HB-DH and LDH in the observation group were (123.54 ±36.57) U/L,(1 786.83 ± 542.37) U/L,(584.63 ± 164.54) U/L,(652.31 ± 187.38) U/L,(956.38 ± 257.64) U/L,respectively,which were significantly higher than those in the control group [(65.38 ± 20.34) U/L,(675.48 ± 240.32) U/L,(48.61 ± 12.15) U/L,(248.36 ± 51.69) U/L,(581.36 ± 102.67) U/L] (t =13.35,17.75,31.50,20.10,13.04,all P ＜0.05).The pH,BE,PaO2,PaCO2 of the observation group were (7.02 ±0.13),(9.56 ± 1.74) mmol/L,(3.26 ± 0.26) kPa,(6.88 ± 1.24) kPa,respectively,which of the control group were (7.20 ± 0.16),(8.96 ± 1.52) mmol/L,(3.21 ±0.33) kPa,(6.58 ± 1.84) kPa,respectively.the pH value in the observation group was significantly lower than that in the control group (t =8.27,P ＜ 0.05).The BE,PaO2 and PaCO2 between the two groups had no statistically significant differences (all P ＞ 0.05).In the observation group,the levels of AST,CK,CK-MB,HB-DH and LDH of the neonates with ideal prognosis were (59.68 ±-13.52) U/L,(542.36 ± 103.65) U/L,(49.37 ± 14.25) U/L,(275.36 ± 64.51) U/L,(567.35 ± 115.24) U/L,respectively,which were significantly lower than those of the neonates with poor prognosis [(81.32 ± 36.24) U/L,(1 265.38 ± 362.74) U/L,(168.35 ±50.01)U/L,(602.31 ± 205.34)U/L,(853.64 ± 212.54)U/L],and the pH value of the observation group with ideal prognosis was significantly higher than that of the neonates with poor prognosis [(7.19 ± 0.21) vs.(7.01 ±0.18)],the differences were statistically significant (t =14.67,14.42,17.22,11.14,7.14,2.91,all P ＜ 0.05).Conclusion The myocardial enzymes and umbilical artery blood gas analysis can be used in the early diagnosis of HIE and evaluation of the prognosis,and it has high clinical value.

OBJECTIVETo explore a feasible method to repair full-thickness skin defects with tissue engineered techniques.

METHODSThe skin specimens were cut from the Changfeng hybrid swines' abdomen, then keratinocytes and fibroblasts were isolated and harvested by trypsin, EDTA and type II collagenase. The cells were seeded in petri dishes for primary culture. When the cells were in logarithmic growth phase, they were treated with dispase II (keratinocytes) or trypsin (fibroblasts) to separate them from the floor of the tissue culture dishes. A biodegradable material-pluronic F-127 was prefabricated and mixed with these cells, and then the cells-pluronic compounds were seeded evenly into polyglycolic acid (PGA). Tinally the constructs were replanted to autologous animals to repair full-thickness skin defects. Histological changes were observed in 1, 2, 4 and 8 weeks postsurgery.

RESULTSThe cells-pluronic F-127-PGA compounds could repair autologous full-thickness skin defects. Histologically, the tissue engineered skin was similar to normal skin with stratified epidermis overlying a moderately thick collageneous dermis.

CONCLUSIONTissue engineered skin can repair autologous full-thickness skin defects with primary-cultured keratinocytes and fibroblasts as seed cells and PGA as a cell carrier.

Animals ; Female ; Fibroblasts ; physiology ; Male ; Polyglycolic Acid ; pharmacology ; Skin Transplantation ; Skin, Artificial ; Swine ; Tissue Engineering

OBJECTIVETo investigate the feasibility of chondrogenic phenotype differentiation of adult swine bone marrow stem cells(MSCs) in a defined medium as seeding cells in cartilage tissue engineering.

METHODSA volume of 5 ml bone marrow was aspirated from swine iliac crest and cultured in the complete medium of DMEM-LG for two weeks. The growth and ultrastructure of the cultured MSCs were observed. Immunohistochemistry and in situ hybridization were applied to detect the expression of collagen type II.

RESULTSThe MSCs changed from a spindle-like fibroblastic appearance to a polygonal shape when transferred from the complete medium of DMEM-LG to a defined medium. A large amount of endoplasmic reticulum was observed in large Golgi ccmplex and mitochondria. The differentiation of MSCs toward chondrogenic phenotype was verified by the positive result of collagen type II through immunohistochemistry and in situ hybridization respectively.

CONCLUSIONSBone marrow stem cells obtained from adult swine can differentiate to be chondrogenic phenotype when cultured in vitro. MSCs can likely be served as optimal autogenous cell source for cartilage tissue engineering.

Animals ; Bone Marrow Cells ; physiology ; ultrastructure ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; physiology ; Collagen Type II ; genetics ; Phenotype ; RNA, Messenger ; analysis ; Stem Cells ; physiology ; ultrastructure ; Swine ; Tissue Engineering ; Transforming Growth Factor beta ; physiology

OBJECTIVETo study transforming growth factor-beta1 (TGF-beta1) autoproduction in keloid fibroblasts and the regulation effect of blocking TGF-beta intracellular signaling on rhTGF-beta1 autoproduction.

METHODSKeloid fibroblasts cultured in vitro were treated with either rhTGF-beta1 (5 ng/ml) or recombinant adenovirus containing a truncated type II TGF-beta receptor gene (50 pfu/cell). Their effects of regulating gene expression of TGF-beta1 and its receptor I and II were observed with Northern blot.

RESULTSrhTGF-beta1 up-regulated the gene expression of TGF-beta1 and receptor I, but not receptor II. Over-expression of the truncated receptor II down-regulated the gene expression of TGF-beta1 and its receptor I, but not receptor II.

CONCLUSIONSTGF-beta1 autoproduction was observed in keloid fibroblasts. Over-expression of the truncated TGFbeta receptor II decreased TGF-beta1 autoproduction via blocking TGF-beta receptor signaling.

Activin Receptors, Type I ; biosynthesis ; pharmacology ; Cells, Cultured ; Down-Regulation ; Fibroblasts ; drug effects ; metabolism ; Gene Expression ; Humans ; Keloid ; metabolism ; Protein-Serine-Threonine Kinases ; RNA, Messenger ; genetics ; metabolism ; Receptors, Transforming Growth Factor beta ; biosynthesis ; metabolism ; Sensitivity and Specificity ; Signal Transduction ; Trans-Activators ; metabolism ; Up-Regulation

Objective: To explore the expression of IQGAP1 (Ras GTPase-activating-like protein containing IQ domain) in esophageal squamous cell carcinoma (ESCC) tissues and cell lines and its effect on the proliferation and invasion of TE-2 cell. Methods: Totally 125 pairs of cancer tissues and para-cancerous tissues from ESCC patients, who underwent surgical resection inAffiliated Tumor Hospital of Zhengzhou University from January 2015 to December 2016, were included in this study; in addition, ESCC cell lines (TE-2, TE3, ECA109) and normal esophageal epithelial cell line Het-1A were also collected. The expression of IQGAP1 was detected by immunohistochemical staining and its relationship with cliniopathological features was also analyzed. IQGAP1 mRNA and protein expressions in ESCC cell lines were detected by Real-time quantitative PCR (qPCR) and Western blotting, respectively. TE-2 cells were transfected with si-IQGAP1 (positive transfection group) and si-CTRL (negative control group) plasmids, and the effects of IQGAP1 silencing on the proliferation and invasion of TE-2 cells were detected by MTT and Transwell assay. The expressions of E-cadherin and Ncadherin were detected by Western blotting. Results: The positive expression rate of IQGAP1 in ESCC tissues was significantly higher than that in para-cancerous tissues (P<0.05), which was closely related to tumor stage and histologic grade (all P<0.05). The mRNAand protein expressions of IQGAP1 in TE-2, TE-3 and ECA109 cells were significantly higher than those in Het-1Acells (all P<0.05).After IQGAP1 was silenced, compared with the negative control group and the blank group, the expression of IQGAP1 mRNAand protein in the positive transfection group significantly decreased (all P<0.05); the proliferation and invasiveness of TE-2 cells significantly decreased (all P<0.05); E-cardherin was up-regulated while N-cardherin was down-regulated (all P<0.05) in the positive interference group. Conclusion: IQGAP1 is highly expressed in ESCC tissues, and si-IQGAP1 can inhibit the proliferation and invasion of TE-2 cells, which plays an important role in the occurrence and development of ESCC.

Objective:To identify the radiomics features related to the occurrence of radiation pneumonitis based on localized CT images of the chest in lung cancer patients, establish a machine learning model and investigate the value of radiomics technology in predicting the incidence of radiation pneumonitis.Methods:Clinical data of 86 patients with stage Ⅲ non-small cell lung cancer who received radical intensity-modulated radiation therapy (IMRT) were retrospectively analyzed. The radiation pneumonitis was graded by follow-up imaging data and clinical information. The planning CT images were collected. The lung was used as the volume of interest for extraction of radiomics features. The radiomics features, clinical and dosimetric parameters associated with the incidence of radiation pneumonitis were analyzed. Using the support vector machine to construct the model, the prediction performance of the model was evaluated by the five-fold verification method.Results:A total of 1029 radiomics features were extracted from CT images and 5 features were selected by ANOVA and LASSO. Two validation sets showed differences between adopting radiomics features alone and incorporating clinical and dosimetric parameters and radiomics features (AUC=0.67 and 0.71, respectively).Conclusions:The radiomics model constructed by planning CT images of lung cancer patients has the potential to predict the occurrence of radiation pneumonitis. Addition of clinical and dosimetric parameters can further improve the prediction performance of the model.