Objective : To analyze the effects of transglutaminase 2(TGM2) gene interference on the malignant proliferation and movement of lung cancer cells . Methods : A549 cells in logarithmic growth phase were divided into control group , shRNA⁃NC group and TGM2⁃shRNA1 group . After transfection , expression levels of TGM2 mRNA A549 cells in logarithmic growth phase were divided into control group , shRNA⁃NC group and TGM2⁃shRNA1 group . After transfection , expression levels of TGM2 mRNA clone formation assay . The proliferation activity of cells was detected by CCK⁃8 . The apoptosis rate was detected by flow cytometer. The invasion of cells was detected by Transwell . The number of tubule formation in each group was observed under microscope . The expression of E ⁃cadherin (E⁃cad) , N ⁃cadherin (N⁃cad) , fibronectin (FN) and vascular endothelial growth factor (VEGF) in each group was detected by Western blot . Results : Compared with shRNA⁃NC group , the mRNA level and protein expression of TGM2 in TGM2⁃shRNA1 group were decreased (P <0. 05) , and the rate of clone formation , proliferation activity , number of invasive cells and number of tubules formation were decreased ( P < 0. 05) . The apoptosis rate and the expression of E ⁃cad protein were increased ( P <0. 05) , while the expression of N ⁃cad , FN and VEGF protein were decreased (P < 0. 05) . Conclusion TGM2 gene silencing can inhibit the expression of VEGF protein , inhibit the malignant proliferation and invasion of A549 cells , and promote apoptosis .

Objective : To analyze the effects of miR⁃346 on alleviating cardiac function damage and oxidative stress in rats undergoing myocardial ischemia/reperfusion (I/R) . Methods : A total of 48 rats were randomly divided into control group , I/R group , I/R + agomiR⁃NC group and I/R + agomiR⁃346 group. I/R model was replicated by the ligation of left anterior descending coronary artery. The recombinant adeno⁃associated virus miR⁃346 was injected through the tail vein for overexpression intervention. After 120 minutes of reperfusion , heart rate (HR) , left ventricular ejection fraction (LVEF) , fractional shortening (FS) and left ventricular wall thickness (LVWT) , levels of serum creatine kinase MB (CK⁃MB) , myoglobin (Mb) , lactate dehydrogenase (LDH) and cardiac troponin I (cTnI) , and levels of superoxide dismutase ( SOD) , glutathion peroxidase ( GSH⁃Px ) and malondialdehyde (MDA) in myocardial tissues were detected. The apoptosis of myocardial tissue cells was detected by TUNEL staining. The expression of B⁃cell lymphoma/leukemia⁃2 (Bcl⁃2) gene , Bcl⁃associated x protein (Bax) , cysteine protease 3 ( Cas⁃3 ) and Cas⁃9 in myocardial tissues was detected by Western blot. Results : HR , LVEF , FS and LVWT in I/R group were lower than those in control group , the levels of serum CK⁃MB , LDH , Mb and cTnI were higher than those in control group , the expression of MDA , Bax/Bcl⁃2 , cleaved Cas⁃3/Cas⁃3 and cleaved Cas⁃9/Cas⁃9 proteins in myocardial tissues was higher than that in control group , the levels of SOD and GSH⁃px were lower than those in control group , and the apoptosis rate of myocardial tissue cells was higher than that in control group (P < 0. 05) . HR , LVEF , FS and LVWT in I/R + agomiR⁃346 group were higher than those in I/R + agomiR⁃NC group , the levels of serum CK⁃MB , LDH , Mb and cTnI were lower than those in I/R + agomiR⁃NC group , the expression of MDA , Bax/Bcl⁃2 , cleaved Cas⁃3/Cas⁃3 and cleaved Cas⁃9/Cas⁃9 proteins in myocardial tissues was lower than that in I/R + agomiR⁃NC group , the levels of SOD and GSH⁃px were higher than those in I/R + agomiR⁃NC group , and the apoptosis rate of myocardial tissue cells was lower than that in I/R + agomiR⁃NC group ( P <0. 05) . Conclusion The miR⁃346 overexpression can reduce oxidative stress level in I/R rats , alleviate myocardial tissue damage , and improve cardiac function.

Cell Line ; Cholesterol ; biosynthesis ; Hepatocytes ; drug effects ; metabolism ; Humans ; Hydroxymethylglutaryl-CoA Synthase ; metabolism ; Lipogenesis ; Quercetin ; pharmacology

OBJECTIVE: To study the relationship between the postmortem interval (PMI) and the changes of DNA content in rabbit's cornea epithelial and endothelium. METHODS: Using the computerized image analysis to measure the DNA content in the cornea epithelial and endothelium of 105 rabbit's at every six hours during 72 hours after death. RESULTS: The degradation rate of DNA in nuclear has an apparent relationship in 72 hours after death of the rabbits. CONCLUSION The determination of the quantity of DNA in cornea nuclear is a new method to estimate the PMI accurately.
Animals ; DNA/analysis* ; Endothelium, Corneal/chemistry* ; Epithelium, Corneal/chemistry* ; Female ; Forensic Medicine ; Male ; Postmortem Changes ; Rabbits ; Signal Processing, Computer-Assisted ; Time Factors

ObjectiveTo investigate the role and mechanism of hyodeoxycholic acid （HDCA） in the progression of metabolic associated fatty liver disease （MAFLD）， and to provide a new theoretical basis for further clarifying the pathogenesis of MAFLD. MethodsL02 hepatocytes were used as experimental cells， and palmitic acid was used to induce steatosis in L02 cells. The farnesoid X receptor （FXR） siRNA interference chain technique was used to construct a hepatocyte cell line with low FXR expression. CCK8 assay was used to observe the effect of HDCA on L02 steatosis hepatocytes at different concentrations （0， 100， 200， 300， and 400 μmol/L） and time points （12， 24， 36， and 48 hours）. The method of qRT-PCR was used to measure the mRNA expression levels of FXR， proliferating cell nuclear antigen （PCNA）， Cyclin D1， phosphatidylinositol 3-kinase （PI3K）， and protein kinase-B （AKT）， and Western blot was used to measure the protein expression levels of FXR， Cyclin D1， PCNA， PI3K， phosphorylated PI3K （p-PI3K）， AKT， and phosphorylated （p-AKT）. A one-way analysis of variance was used for comparison of normally distributed continuous data with homogeneity of variance between multiple groups， and the Tukey HSD test was used for further comparison between two groups； the Welch analysis of variance was used for comparison of normally distributed continuous data with heterogeneity of variance between multiple groups， and the Games-Howell test was used for further comparison between two groups. The independent-samples t test was used for comparison between two groups. ResultsCCK8 assay showed a significant reduction in the viability of L02 cells and steatosis hepatocytes treated by 300 μmol/L HDCA （P<0.05）， and qRT-PCR showed a significant increase in the mRNA expression level of FXR and significant reductions in the mRNA expression levels of PCNA， Cyclin D1， PI3K， and AKT （all P<0.05）. Western blot showed a significant increase in the protein expression level of FRX （P<0.05）， and after interference of FXR expression in L02 cells， there were significant increases in the protein expression levels of PCNA， PI3K， p-PI3K， AKT， and p-AKT （all P<0.05）. ConclusionHDCA inhibits the PI3K/AKT signaling pathway by upregulating FXR expression， thereby inducing a reduction in the viability of steatosis hepatocytes.

OBJECTIVEThis clinical study was to improve the surgical treatment to craniomaxillofacial tissue defects.

METHODSSince 1997, eight cases with severe craniomaxillofacial defects were treated using free latissimus dorsi myocutaneous flaps. In the operation, nerve anastomosis was performed. Of the 8 cases, 7 were treated in one stage, 1 was treated in 3 steps. The craniomaxillofacial defects ranged from 10 cm x 8 cm to 30 cm x 12 cm. The flaps was 12 cm x 10 cm to 32 cm x 16 cm in size.

RESULTSPostoperative follow-up for 6 months to 4 years demonstrated satisfactory results in all the cases. There was neither necrosis nor ulcer after the operation. The sensation recovery of the flap was also satisfactory.

CONCLUSIONFree transfer of the latissimus dorsi myocutaneous flap is an ideal treatment to severe craniomaxillofacial defects as it possesses the advantages of reliable blood supply, ability against infections, large size, concealed donor site, and functional restoration of sensation and movement.

Adult ; Aged ; Craniofacial Abnormalities ; surgery ; Female ; Humans ; Male ; Maxillofacial Abnormalities ; surgery ; Middle Aged ; Muscles ; metabolism ; surgery ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; Transplantation, Homologous ; Treatment Outcome

OBJECTIVETo observe the therapeutic effect of Xiaobai Mixture (XBM) in treating vitiligo.

METHODSSeventy-four patients with vitiligo were randomly divided into the XBM group treated with XBM and the control group treated with 8-MOP. The therapeutic effect, nail-fold microcirculation, plasma endothelin-1, serum immunoglobulin were observed and compared.

RESULTSThe therapeutic effect of XBM was better than that of 8-MOP (P < 0.05). XBM could also obviously improve the nail-fold microcirculation, elevate the plasma endothelin-1 level and lower the serum IgG (P < 0.01).

CONCLUSIONXBM has superiority in treating vitiligo.

Adolescent ; Adult ; Drugs, Chinese Herbal ; therapeutic use ; Endothelin-1 ; blood ; Female ; Humans ; Immunoglobulin G ; blood ; Male ; Microcirculation ; Middle Aged ; Nails ; blood supply ; Phytotherapy ; Vitiligo ; drug therapy

To explore the effect of sinomenine on the nitric oxide (NO)/neural nitric oxide synthase (nNOS) system in the cerebellum and spinal cord of morphine-dependent and morphine-withdrawal Kunming mice, mice were subjected to injection of morphine with an increasing dose for 5 d, and then were treated with sinomenine (40 mg/kg, i.p.) for another 5 d. Naloxone was used to develop acute withdrawal, and the withdrawal syndromes, including teeth chattering, twisting, straightening, sneezing and ptosis, were investigated. nNOS mRNA expressions in the cerebellum and spinal cord were determined by semi-quantitative RT-PCR. nNOS activity and NO level were determined by the chemistry-colorimetry and nitrate reductase-reduction, respectively. The results obtained were as follows: (1) Sinomenine restored the decrease in body weight and alleviated the signs of morphine-withdrawal in mice. (2) Sinomenine also reduced the increases in nNOS mRNA expression and nNOS activity resulting from morphine-dependence, and simultaneously attenuated the high level of NO in both tissues following morphine-withdrawal. (3) Administration of sinomenine alone did not develop physical dependence in mice. The results obtained indicate that sinomenine may attenuate morphine addiction and significantly alleviate morphine-withdrawal symptoms, and the mechanism may be associated with the effect of sinomenine on the NO/nNOS system in the cerebellum and spinal cord.
Animals ; Body Weight ; drug effects ; Cerebellum ; drug effects ; metabolism ; Male ; Mice ; Morphinans ; pharmacology ; therapeutic use ; Nitric Oxide ; analysis ; Nitric Oxide Synthase Type I ; genetics ; metabolism ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; Spinal Cord ; drug effects ; metabolism ; Substance Withdrawal Syndrome ; drug therapy ; metabolism

OBJECTIVETo study the features of vascular smooth muscle cell (VSMC) proliferation induced by endothelin-1 (ET-1).

METHODSVSMCs of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats were cultured and treated with ET-1. Basic fibroblast growth factor (bFGF) gene expression was measured using both Northern blot and an enzyme-linked immunoassay.

RESULTSET-1 resulted in an increase in bFGF transcripts at 8 - 24 h; bFGF levels were significantly higher in VSMCs treated with ET-1 than in those not treated. However, VSMCs growth responses in SHR and WKY were different. Smooth muscle cells of SHR were hyper-responsive to ET-1. Maximal bFGF mRNA levels were elevated 3.5-fold at 4 h of stimulation in WKY and 8-fold at 8h in SHR4. Moreover, the proliferation of VSMCs induced by ET-1 was inhibited by antisense phosphorothioate oligodeoxynucleotides (10 micromol/L AS-bFGF) but not sense bFGF oligomers at the same concentrations, being reduced by 80% in SHR and 40% in WKY vs control, respectively. Furthermore, the effect of AS-bFGF oligomers on SHR SMC proliferation is significantly greater than on WKY SMC proliferation.

CONCLUSIONET-1 may be required for exaggerated vascular growth responses in SHR and bFGF may be involved.

Animals ; Cell Division ; drug effects ; Cells, Cultured ; DNA, Antisense ; pharmacology ; Dose-Response Relationship, Drug ; Endothelin-1 ; pharmacology ; Fibroblast Growth Factor 2 ; genetics ; Gene Expression Regulation ; drug effects ; Muscle, Smooth, Vascular ; cytology ; drug effects ; metabolism ; Oligonucleotides ; pharmacology ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Time Factors

In order to determine the involvement of CALM-AF10 fusion transcripted in primary leukaemias with t(10;11) and its chemotherapy sensitivity in vitro, the AF10-CALM fusion transcripts were detected by reverse transcription-polymerase chain reaction (RT-PCR), and the chemotherapy sensitivity testing in vitro was undergone by MTT assay in five t(10;11) leukemia samples from patients with ALL, AML and lymphoblastic lymphoma. The results showed that five different-sized AF10-CALM product and four different-sized CALM-AF10 products were detected. The chemotherapy sensitivity of leukemic cells with t(10;11) in vitro to drugs is lower than that of leukemic cells without t(10;11). 3 out of 5 cases of t(10;11) leukemia were sensitive to chemotherapeutic drugs, while 31 out of 36 cases of leukemia without t(10;11) were sensitive at same condition. There were significant differences (P < 0.01), consistent with clinical features of patients. Apoptosis rate of leukemic cells with t(10;11) induced by chemotherapeutic drugs was lower than that of leukemic cells without t(10;11), (16.37 +/- 2.56)%, and (33.75 +/- 5.59)%, respectively (P < 0.01). It is concluded that the CALM-AF10 fusion transcripts are a common features and are involved in the pathogenesis of haematological malignancies with t(10;11), and are associated with a poor prognosis.
Antineoplastic Agents ; pharmacology ; Cell Survival ; drug effects ; Chromosomes, Human, Pair 10 ; genetics ; Chromosomes, Human, Pair 11 ; genetics ; Humans ; Leukemia ; genetics ; pathology ; Oncogene Proteins, Fusion ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transcriptional Activation ; drug effects ; Translocation, Genetic ; Tumor Cells, Cultured