OBJECTIVE To investigate the apoptosis-inducing effect of esculetin （Esc） on acute myeloid leukemia （AML） HL-60 cells by regulating the protein kinase B （AKT）/S-phase kinase-associated protein 2 （SKP2）/MutT homolog 1 （MTH1） pathway. METHODS AML HL-60 cells were randomly divided into control group （routine culture）， Esc low-concentration group （L-Esc group， 25 μmol/L Esc）， Esc medium-concentration group （M-Esc group， 50 μmol/L Esc）， Esc high-concentration group （H-Esc group， 100 μmol/L Esc）， and high-concentration of Esc+ SC79 （AKT agonist） group （100 μmol/L Esc+5 μmol/L SC79）. Cell proliferation in each group was detected by MTT assay and colony formation assay. The level of reactive oxygen species （ROS） in cells was measured by using the CM-H2DCFDA fluorescent probe. Cell apoptosis was analyzed by flow cytometry. Western blot assay was performed to detect the expression levels of apoptosis-related proteins [B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， cleaved caspase-3]， AKT/SKP2/MTH1 pathway-related proteins （p-AKT， AKT， SKP2， MTH1）， along with the upstream and downstream proteins of AKT phosphatidylinositol 3-kinase （PI3K）， cyclin-dependent kinase inhibitor 1 （P21） and cyclin-dependent kinase inhibitor 1B （P27）. RESULTS Compared with control group， the cell viability， colony number， and the phosphorylation levels of AKT and PI3K proteins as well as protein expressions of SKP2， MTH1 and Bcl-2 were significantly decreased （P＜0.05）， while ROS level， apoptosis rate， and the expression levels of Bax， cleaved caspase-3， P21 and P27 proteins were significantly increased （P＜0.05）. Moreover， the effects of Esc exhibited concentration-dependence （P＜0.05）. Compared with H-Esc group， above indexes of high-concentration of Esc+ SC79 group were reversed significantly （P＜0.05）. CONCLUSIONS Esc may promote massive ROS production and induce activation of apoptosis in HL-60 cells by inhibiting the AKT/SKP2/MTH1 pathway， thus inhibiting the proliferation of HL-60 cells.

Objective: Screen the pathogenic genes of a pedigree with clinical manifestation of familial dilated cardiomyopathy in Inner Mongolia. Methods: A total of 3 patients with dilated cardiomyopathy and 20 family members from the same family were examined in Ordos Central Hospital in Inner Mongolia from October, 2003 to August, 2017. Data on medical history, physical examinations, electrocardiograms, and echocardiography were obtained. 5 ml peripheral blood was sampled for per person. Chip Capture Sequencing technology was used to capture all the exons and splice sites of the genes that associated with hereditary cardiomyopathy and hereditary arrhythmia. The mutations in these genes were detected by high-throughput sequencing. All suspected pathogenic loci identified by high-throughput sequencing were verified by Sanger sequencing used for mutation detection. One hundred and fifty gender, age and race matched healthy people were included as the control group. Results: Pathogenic gene variations were detected in 3 symptomatic family members and 1 carrier from the pedigree. Five pathogenic gene variations were identified in the proband (Ⅱ1), a pSer236Gly and a pArg215Cys variation in the MYBPC3 gene, a pGln90Arg variation in the DSP gene, and pAsn2912Asp and pGlu2910Val variation in the DMD gene. One pathogenic variation was detected in Ⅲ3, which was a pArg215Cys variation in the MYBPC3 gene. Two pathogenic variations were detected in Ⅲ7, a pSer236Gly variation in the MYBPC3 gene and a pGln90Arg variation in the DSP gene. Two pathogenic variations were detected in the Ⅳ7, a pSer236Gly variation in the MYBPC3 gene and a pGln90Arg variation in the DSP gene. No gene variation loci were detected in the other family members and the control group. Conclusion MYBPC3 gene, DSP gene and DMD gene variations are present in the familial dilated cardiomyopathy pedigree from Inner Mongolia, and these variations may be related with familial dilated cardiomyopathy.

Objective To investigate the value of endoscopic retrograde appendicitis therapy ( ERAT) in the diagnosis and treatment of atypical acute appendicitis. Methods All the 48 patients suspected of atypical acute appendicitis in Jiangsu Province Hospital from January 2015 to December 2016 were randomly divided into ERAT group and conservative treatment group according to the treatment method. The final appendectomy rate of the two groups was analyzed. Results Only 17 of the 24 patients in the ERAT group received endoscopic treatment because of complex conditions or personal wishes, and 16 cases were diagnosed as acute appendicitis. Surgical resection was performed in 5 cases because of recurrence of the disease after ERAT, and the appendectomy rate was 31. 2％ ( 5/16 ) . In the conservative treatment group, all 24 patients were treated with antibiotics. Twenty of them underwent surgical resection with appendectomy rate of 83. 3％ ( 20/24) , and 1 of them had appendiceal perforation. The appendectomy rate of the ERAT group was significantly lower than that of the conservative treatment group (χ2=11. 111, P<0. 05) . Conclusion ERAT has a high diagnostic and therapeutic value for atypical acute appendicitis.

Aim Toinvestigatewhetherexposureto Sanguinarine (SAN ) can inhibit cell proliferation in human mammary adenocarcinoma cells (MCF-7 ) and thepossiblemechanism.Methods WeexposedMCF-7 to anticancer compound SAN,cell viability was as-sessed by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT ) reduction assay. ROS was measured using confocal microscopy,expres-sion of caspase-3 ,caspase-8 and caspase-9 were calcu-latedusingchemiluminescencemethod.Results SAN remarkably inhibited growth of human mammary adeno-carcinoma MCF-7 cells by decreasing cell proliferation. ROS release and caspase-3,caspase-8,caspase-9 ex-pression were stimulated by SAN in MCF-7 ,and these changes were abolished by the antioxidant,N-acetyl-cysteine(NAC).Conclusion Regulationofcaspases expression and release from MCF-7 cells are possibly e-voked by SAN through reactive oxygen species.

Objective To investigate the role of quality care to improve the non-small cell lung cancer related fatigue in patients with cancer and quality of life. Methods A total of 110 cases of non-small cell lung cancer patients were randomly divided into intervention group and control group,each of 55 cases, two groups were given conventional can-cer chemotherapy treatments, the intervention group received quality care interventions on this basis, used self-assess-ment amendment Piper fatigue scale (RPFS), Anderson symptom assessment scale (MDASI), the quality of life of cancer patients before and after assessment scale(EORTC QLQ-C30) evaluated two interventions state. Results The differences of emotional, cognitive and RPFS scores and intervention were statistically significant between before and after inter-vention in the intervention group(P<0.05), and compared with the control group, the differences were statistically sig nificant (P<0.05), the differences of MDASI scale fatigue, sleep, nausea, anguish, sadness, numbness, general activities, relationships, enjoyment of life scores between the intervention group and control group after the intervention, were sta-tistically significant (P<0.05). The differences of EORTC QLQ-C30 scale physical function, role function, emotional function, cognitive function, social function, overall health score between intervention group and control group after in-tervention were statistically significant (P<0.05). Conclusion Quality care could relieve patients with non-small cell lung cancer-related fatigue and improve cancer-related symptoms and quality of life.

BACKGROUNDImplanted medical catheter-related infections are increasing, hence a need for developing catheter polymers bonded to antimicrobials. We evaluated preventive effects of levofloxacin-impregnated catheters in catheter-related Psuedomonas aeruginosa (strain PAO1) infection.

METHODSDrug release from levofloxacin-impregnated catheters was measured in vitro. Levofloxacin-impregnated catheters and polyvinyl chloride (PVC) catheters were immersed in 5 ml 50% Luria Bertani medium containing 10(8) CFU/ml Pseudomonas aeruginosa then incubated for 6, 12, 24 or 48 hours at 37°C when bacteria adhering to the catheters and bacteria in the growth culture medium were determined. Impregnated and PVC catheters were singly implanted subcutaneously in mice, 50 µl (10(7)CFU) of PAO1 was injected into catheters. After the first and fifth days challenge, bacterial counts on implanted catheters and in surrounding tissues were determined microbiologically. Bacterial colonization and biofilm formation on implanted catheters were assessed by scanning electron microscopy.

RESULTSDrug release from levofloxacin-impregnated catheters was rapid. Levofloxacin-impregnated catheters had significantly fewer bacteria compared to PVC in vitro. After first and fifth day of challenge, no or significantly fewer bacteria adhered to impregnated catheters or in surrounding tissues compared to PVC. Scanning electron microscopical images after first day displayed from none to significantly fewer bacteria adhering to impregnated implanted catheters, compared to bacteria and microcolonies adhering to PVC catheters. After the fifth day, no bacteria were found on impregnated catheters, compared to clusters surrounding mucus-like substance and coral-shaped biofilms with polymorphonuclear leukocyte on PVC catheters. After the first day of challenge, secretion occurred in all implanted catheters with surrounding tissues mildly hyperaemic and swollen. After the fifth day, minute secretions inside impregnated catheters and no inflammation in tissues, whereas purulent secretion inside PVC catheters and abscesses in surrounding tissues.

CONCLUSIONLevofloxacin-impregnated catheter is a promising new strategy for prevention of catheter-related Psuedomonas aeruginosa infection.

Animals ; Biofilms ; drug effects ; Catheters, Indwelling ; microbiology ; Female ; Levofloxacin ; therapeutic use ; Mice ; Pseudomonas Infections ; prevention & control ; Pseudomonas aeruginosa ; pathogenicity

Objective To observe the destructive and scavenging effect of ambroxol (AMB) on the biofilm (BF) of Pseudomonas aeruginosa (P.a).To evaluate the synergistically bactericidal effect of AMB along with levofloxacin (LFX) on BF of P.a.Methods The early model (cultured for 3 d) and mature model (cultured for 7 d) of P.a wild strain (PAO1) BF were established,in vitro,respectively.The models were randomly (random number) divided into control group,AMB group and AMB + LFX group.The concentrations of AMB were 256 μg/ml and 512 μg/ml,respectively.When the early BF model and mature BF model were made,different concentrations of AMB were added in AMB group and AMB + LFX (1μg/ml) was added in AMB + LFX group.The number of viable P.a on the BF carrier was counted with the continuous dilution method 24 h after AMB or/and LFX added.Then,the BF morphological changes on the carrier surface were observed by using scanning electron microscopy (SEM).Measured data were analyzed with single factor analysis of variance (One-Way ANOVA).Results Both in the early BF model and in the mature BF model,the SEM examination showed that the BF in AMB group was significantly reduced compared to the control group,and this reduction of BF was in dose-dependent manner.LFX 1 μg/ml could reduce the number of viable bacterial in BF in both early model and mature model (P ＜ 0.05).LFX with addition of different concentrations of AMB showed stronger bactericidal effect than LFX used alone identified by more significant reduction in the number of colonv within the BF (P ＜ 0.05).Furthermore,the LFX combined with 512 μg/ml AMB reduced more significant number of colony apparently than the LFX combined with 256 μg/ml AMB (P ＜ 0.05).Conclusions AMB can destroy the early BF or mature BF partly,and LFX alone can partly reduce the number of viable P.a within BF.When LFX combined with AMB,they exert a synergistically bactericidal effect.

Objective To evaluate the effect of azithromycin on Pseudomonas aeruginosa PAO1 biofilm formation and virulence factors production. Methods Detect the minimum inhibitory concentration of azithromycin against PAO1 by 2-fold dilution method. Crystal violet staining assay was used for initial adhesion assays. The PAO1 biofilm was established in vitro and observed by scanning electron microscope. Viable bacterial counts were determined by serial dilution. LasB elastolytic activity was determined by using Elastin-Congo Red. Protease activity was determined by Azo-casein. Chloroform extraction method was used for pyoverdine assay . The orcinol assay was used to directly assess the amount of rhamnolipids . Results Scanning electron microscope biofilm and viable bacterial counts of PAO1 adhered to the surface of catheter in PAO1 azithromycin group were less than the PAO1 control group after incubated for 3 d and 7 d ( P ＜0.05), and the initial adhesion was weaker ( P ＜ 0. 05 ). The virulence factors production were obviously decreased (P ＜0.01 ). LasB elastolytic activity and pyoverdine were even reduced to the same as with the PA-JP3 group ( P ＞ 0.05 ), but the protease activity and the rhamnolipids concentration were higher than the PA-JP3 group ( P ＜ 0.05 ). Conclusion Azithromycin can inhibit PAO1 bioflim formation in vitro and virulence factors production.

Objective To investigate the effects of caveolin-1 overexpressing on the growth of cervical squamous cell cancer Hela cell line. Methods Eukaryotic expression vector of human caveolin-1 gene was introduced into Hela cells by Lipofectamine. The clones stably overexpressed caveolin-1 were identiffed by RT-PCR, immunofluorescence cell staining techniques and Westernblotting. Cells proliferation viabihty was tested by MTT assay, and flow cytometry was used to assay the cell cycle and apoptosis, and the relative phosphorylation level of extracellular regulated protein kinases (Erk1/2) were detected by Westernblotting. Results The clones stably overexpressed caveolin-1 were obtained. Compared with the parental Hela cells, the tranfected cells exhibited a slower rate of growth. FAGS analysis results revealed that overexpression of caveolin-1 resulted in the cell cycle arrest in the G0/G1 [ ( 68. 04 ± 2. 57 ) % vs ( 53.41 ±1.01)%] phase and increased the apoptotic cell fraction[ (19. 18 ±2.20)% vs (5.63 ±0.55)%, P <0. 05 ]. Western blotting results showed that overexpression of caveolin-1 reduced the phosphorylation of Erk1/2(0.28 ±0.05 vs 0.81 ±0.07, P <0.05). Conclusions Overexpression of caveolin-1 suppressed the growth of Hela cells and induced apoptosis, down-regulation of Erk1/2 phosphorylation might be involved in its mechanism.

Objective To explore the expression of KAI1/CD82,E-cadherin and β-catenin in endometrial carcinoma,and to investigate their correlations to clinicopathological parameters of endometrial carcinoma. Methods The expressions of KAI1/CD82,E-cadherin and β-catenin in 76 specimens of endometrial carcinoma,15 specimens of atypical endometrial hyperplasia and 20 specimens of proliferative endometrium were examined by immunohistochemical envision technique.Their correlations to clinicopathological parameters of endometrial carcinoma were statistically analyzed. Results Compare to normal proliferative phase endometrium and atypical endometrial hyperplasia,the expression of KAI1/CD82 in endometrial carcinoma was significantly decreased(P ＜0.01),the abnormal expression of E-cadherin and β-catenin in endometrial carcinoma were significantly higher(all P <0.01).In endometrial carcinoma,the expression of KAI1/CD82 was negative correlated with histological grade and depth of myometrial invasion(P <0.01,P <0.05); The abnormal expression of the E-cadherin is related to histological grade and type(P <0.01,P<0.05); The abnormal expression of β-catenin was positively correlated with histological grade and FIGO stage(P ＜0.01 ,P <0.05).The down-regulation expression of KAI1/CD82 was closely associated with the abnormal expression of E-cadherin and beta-catenin in endometrial carcinoma(P <0.01,P <0.05). Conclusion The down-regulation of KAI1/CD82 and the aberrant expression of E-cadherin and β-catenin could be involved in the development of endometrial carcinoma.The loss or reduced expression of KAI1/CD82 was closely associated with the abnormal expression of E-cadherin and β-catenin in endometrial carcinoma.