3.Effect and Underlying Mechanism of Piperlongumine on Proliferation and Apoptosis of Human Gastric Cancer Cell Line MKN45
Chaoqin DUAN ; Chao DENG ; Xiaoping ZOU
Chinese Journal of Gastroenterology 2016;21(2):69-74
Background:Recently,studies have shown that piperlongumine( PL)selectively killed cancer cells by elevating reactive oxygen species(ROS)in various cancers. However,the effect of PL on gastric cancer cells remained to be further studied. Aims:To investigate the effect of PL on proliferation and apoptosis of human gastric cancer cell line MKN45 and its underlying mechanism. Methods:MKN45 cells were treated with different doses of PL,caspase inhibitor,antioxidant, and their combinations,respectively. Cell viability was assessed by CCK-8 assay;cell cycle,apoptosis and intracellular ROS level were measured by flow cytometry;and Western blotting was employed to determine the expression of apoptosis-related proteins( XIAP,cleaved-caspase3,7,9 and cleaved-PARP),p53 and its downstream target genes( p21, GADD45α and PUMA). Results:PL inhibited the proliferation of MKN45 cells in a dose- and time-dependent manner. In MKN45 cells treated with PL,the proportion of cells in G1 phase,apoptotic rate and intracellular ROS level were significantly increased,the expression of inhibitor of apoptosis protein XIAP was down-regulated,and the caspase-dependent apoptosis pathway,p53 and its downstream target genes were activated. Pretreatment with antioxidant NAC or Z-VAD-FMK, a general caspase inhibitor could partially abolish the effect of PL on ROS production and its antitumor effect. Conclusions:PL can inhibit cell proliferation and induce cell cycle G1 phase arrest and apoptosis in MKN45 cells. Its antitumor effect may be associated with a ROS-mediated p53 activation and subsequent triggering of caspases cascade of cell apoptosis.
4.Research on clinical effect observation of TACE combined with percutaneous microwave coagulation therapy on treatment of patients with hepatocellular carcinoma
Bin LI ; Chao SUN ; Huiyong DUAN
China Medical Equipment 2015;(12):137-139
Objective:To explore the effect of transcatheter arterial chemoembolization (TACE) combined with percutaneous microwave coagulation therapy(PMCT) on treatment of patients with hepatic carcinoma.Methods: The clinical data of 60 cases of liver cancer patients in our hospital in TACE combined with PMCT were analyzed. Results:The operation was successfully performed, no serious complications and death. Efficacy: 4 CR cases, 38 PR cases, 16 SD cases, 2 PD cases, the total efficiency of treatment was 70.0%; After 1, 2, 3 year, survival cases (ratio) were respectively: 46(76.7%), 35(58.3%), 31(51.7%); The levels of Serum alpha fetoprotein (AFP) after operation was significantly reduced, after treatment, the patient's liver function improved significantly; The diameter of the lesions was significantly reduced.Conclusion: TACE combined with PMCT is an effective therapy for hepatic carcinoma.
5.Eosinophil could predict the prognosis of patients with bloodstream infection: a retrospective analysis of 305 cases
Chinese Critical Care Medicine 2017;29(6):496-500
Objective To investigate the value of peripheral blood for the prognosis of patients withbloodstream infection. Methods A retrospective analysis of patients with bloodstream infection was conducted inthe intensive care unit (ICU) of Mianyang Central Hospital of Sichuan from January 2012 to October 2016. Accordingto the 28-day survival, the patients were divided into survival group and death group. The white blood cell (WBC),neutrophils count (NEU), lymphocyte count (LYM), neutrophil/lymphocyte ratio (NLR), monocyte count (MO), eosinophilcount (EO), basophil count (BA), hemoglobin (Hb), platelet count (PLT) and procalcitonin (PCT) in peripheral bloodwere recorded when patients were diagnosed with blood infection. Receiver operating characteristic curve (ROC),Kaplan-Meier survival analysis and Cox regression were used to evaluate the value of these risk factors for predictingthe outcome. Results 305 patients were enrolled. 182 patients survived while 123 patients died during the 28-dayperiod. ① There was no significant difference in gender, age and comorbidities between the two groups. There was nosignificant difference in infection rate between the two groups except for fungal infection rate. The fungal infection ratein the death group was significantly higher than that in the survival group (9.8% vs. 3.3%, P = 0.019). ② The LYM,MO, EO and PLT in the death group were significantly lower than those in the survival group [LYM (×109/L):0.58 (0.29, 0.93) vs. 0.76 (0.44, 1.23), MO (×109/L): 0.47 (0.19, 0.80) vs. 0.58 (0.30, 0.94), EO (×109/L):0.00 (0.00, 0.01) vs. 0.03 (0.01, 0.09), PLT (×1012/L): 89 (47, 148) vs. 126 (82, 186), all P < 0.05]. The NLR in the death group was significantly higher than that in the survival group [17.09 (7.60, 33.51) vs. 12.86 (6.51, 24.85), P < 0.05]. There was no significant difference in the WBC, NEU, BA, Hb and PCT between the two groups. ③ It was shown by ROC curve analysis that the maximum area under the ROC curve (AUC) of EO was 0.755. When the best cut-off value was 0.015×109/L as a predictor of death in 28 days, the sensitivity was 80.3%, and specificity was 64.7%. ④ It was shown by survival analysis that the 28-day survival rate in the patients with EO < 0.015×109/L was significantly lower than that of patients with EO > 0.015×109/L [38.3% (62/162) vs. 83.9% (120/143), χ 2 = 56.999, P = 0.000]. ⑤ It was shown by Cox regression that EO was the independent factor for 28-day survival (β = 1.466, χ 2 = 39.535, P = 0.000). Risk of death was 4.331 times greater in patients with EO < 0.015×109/L than in those with EO > 0.015×109/L [hazard ratio (HR) = 4.331, 95% confidence interval (95%CI) = 2.743-6.840]. Conclusions Compared to other parameters in peripheral blood, EO has the best correlation with the prognosis of bloodstream infection. EO is the independent prognostic predictor for 28-day survival.
6.Fenofibrate inhibits production of reactive oxygen species and oncofetal fibronectin induced by short-term intermittent high glucose through blockade of local aldosterone system in human mesangial cells
Chao DU ; Bo ZHOU ; Yaqian DUAN ; Hong SU
Chinese Journal of Endocrinology and Metabolism 2012;28(8):654-660
Objective To investigate the role of local aldosterone system in oncofetal fibronectin ( oncofetal FN) mRNA expression and reactive oxygen species (ROS) production in human mesangial cells (HMCs) exposed to short-term intermittent high glucose and the effect of Fenofibrate.Methods The HMCs were divided into 8 groups:normal glucose(NG) ;osmotic fluctuation(OF) ;mean glucose load (MGL) ;stable high glucose (SHG),short-term intermittent high glucose (IHG) ; intermittent high glucose plus eplerenone (IHGE) ; intermittent high glucose plus fenofibrate(IHGF) ; and normal glucose plus fenofibrate (NGF) groups.The mRNA expression levels of Aldosterone synthase ( CYP11 B2 ),11 β-hydroxysteroid dehydrogenase type 2 ( 11βHSD2 ) and oncofetal FN were determined by RT-PCR.The expression of CYP11B2 protein was determined by western-blot.Aldosterone level in cell culture supernatant was detected by radioimmunoassay.The expression and translocation of mineralocorticoid receptor (MR)protein were assayed with confocal laser scanning microscopy. ROS levels were determined by Fluorescence microscopy and fluorescence microplate reader.Results ( 1 ) MGL,SHG,and IHG groups showed a 2.41,3.63,and 4.45 times increase in CYP11B2 mRNA expression,and a 1.83,2.15,and 2.78 times increase in CYP11B2 protein expression,respectively,compared with NG group (P < 0.05 ).The aldosterone levels of HMCs culture supernatant in MGL,SHG,and IHG groups were also increased,being 1.49,2.04,and 2.54 times of that in NG group ( P<0.05 ),and the degree of elevation in IHG group was more marked than that in SHG group( P<0.05 ).MR was activated and translocated from cytosol to nucleus in MGL,SHG,and IHG groups.Quantitative analysis showed the ratioes of cytosol/nucleus fluorescence intensity in MGL,SHG,and IHG groups were 15%,38%,and 53% decreased as compared with that in NG group,and the decrease was more marked in IHG group ( P<0.05 ).(2) Oncofetal FN mRNA expression and ROS levels in MGL,SHG,and IHG groups were increased,being 1.54,2.31,3.65 and 1.26,1.91,2.48 times of those in NG group,respectively ( P<0.05 ),and this increase was more marked in group IHG ( P<O.05 ).Compared with IHG group,oncofetal FN mRNA expression and ROS levels in group IHGE were significantly decreased by 54% and 53%,and in group IHGF by 45% and 39%. ( 3 ) CYP11B2 mRNA,protein,and aldosterone levels in IHGF group were decreased by 74%,59%,and 50%,and the activation of MR in group IHGF was inhibited when the ratio of cytosol/nuclear fluorescence intensity was increased 1.88 fold as compared with that in group IHG ( P<0.05 ).Conclusions Increased expressions of oncofetal FN and ROS by HMCs induced by short-term intermittent high glucose were nore marked than those induced by stable high glucose.The mechanism was associated with activation of local aldosterone system.Fenofibrate may inhibit the activation of local aldosterone system and alleviate the injury to HMCs induced by intermittent high glucose.
7.Clinical analysis of 29 hyperthyroid patients combined with cytopenia
Xiaolan ZHANG ; Yu DUAN ; Li ZHANG ; Chao LIU
Chinese Journal of General Practitioners 2008;7(6):412-413
This report was to retrospectively analyze clinical data of 29 hyperthyroid patients with concurrent hematocytopenia.Our findings demonstrated that 5 patients developed cytopenia without any antithyroidism drugs(ATDs),while the other 24 cytopenia patients were only found after taking ATD.In the drug intervention group,17 patients received methimazole(MMI)and 7 took propylthiouracil(PTU).Most cytopenia was dose-dependent and occurred within the first 6 months of medication.Twenty subjects received radioiodine therapy,with no impact on blood cells.We suggested that cytopenia might occur at the early stage of ATD therapy and be related with drug and its dose.Blood cell amount should be monitored,especially for initial ATD therapy.As for interaction between PTU and MMI,alternative radioiodine therapy might seem to be relatively safe.
8.Clinical study on ductoscopic flushing in the treatment of early lactation acute mastitis
Baoyun WANG ; Xiaodong DUAN ; Haiquan ZHANG ; Chao HAN ; Hongwei YAN
Chinese Journal of Primary Medicine and Pharmacy 2017;24(6):905-908
Objective To investigate the clinical application value of ductoscopic flushing in the treatment of early lactation acute mastitis.Methods 52 patients with early acute mammitis were divided into observation group (27 cases)and control group(25 cases)according to the principle of completely random.In the observation group, the patients were checked under the intervention of mammary duct firstly,and then the disease regions of the breast were flushed using ductoscope.In the control group,the patients were treated with artificial breast -milk.The two groups were all treated with the same antibiotics.The curative effect of the two groups was observed,and the statistical analysis was performed.Results In the observation group,the mass extinction time,pain relief time,pyretolysis time, hemogram recovery time,contralateral continued breast -feeding proportion,the proportion of abscess formation,the proportion of back -milk,the proportion of ipsilateral quadrant recurrence were (3.5 ±1.2)h,(5.0 ±0.9)h, (1.0 ±0.1)d,(1.0 ±0.3)d,92.6%,7.4%,7.4%,0.0% respectively,those in the control group were (24.0 ± 3.2)h,(2.0 ±2.1)h,(2.0 ±0.2)d,(3.0 ±0.3)d,88.0%,12.0%,12.0%,8.0% respectively.The differences between the two groups were statistically significant(t =1.72,0.36,0.43,0.72,χ2 =1.83,2.02,1.56,0.34,all P <0.05).Conclusion Ductoscopic flushing has good effect in the treatment of early lactation acute mastitis,and it is worthy of clinical promotion.
9.Prognostic value of procalcitonin in patients with acute paraquat intoxication
Xiaofan ZHANG ; Li LI ; Chao LAN ; Li HUANG ; Guoyu DUAN
Chinese Journal of Emergency Medicine 2016;25(8):1044-1048
Objective To study the prognostic value of procalcitonin (PCT) level in the outcome of patients with paraquat poisoning (PQ).Methods The clinical data of 128 patients with acute PQ admitted to emergency department were collected from March 2013 through March 2014.The patients were divided into two groups:the death group and the survival group (survival of 28 days).Poisoning doses,urine concentration of PQ,time elapsed from poisoning to admission,and time elapsed from poisoning to gastrolavage were documented.And on the 1 st day,the 3rd day and the 7th day after poisoning,serum PCT were detected.The level of PCT was used to investigate the prognostic values in patients with acute PQ in the death group and survival group.Results Of 128 cases,72 (56.3%) survived and 56 died in 28 days.Among them,the level of PCT increased to some extent in the first day in 90 cases,and 48 patients died.According to trend analysis,the levels of PCT in death group on the 1st day,the 3rd day and the 7th day after PQ were significantly higher than those in survival group [ld:(0.96 ±0.13) vs.(0.08 ±0.01),3d:(1.12 ±0.14) vs.(0.28 ±0.05),7d:(1.22 ±0.14) vs.(0.20 ±0.03),P <0.01].There was a trend of escalating PCT levels in death group,whereas the PCT level reached the peak on the 3st day and decreased gradually in the following days in survival group.The early PCT level was obviously related to poisoning doses,urine concentration,CRP,WBC,ALT,CR (the coefficient of association were 0.794,0.723,0.724,0.332,0.700,0.414,respectively,P<0.01).Conclusions The serum level of PCT increased in patients with acute PQ was significantly positively correlated with the oral dose and urine concentration of paraquat,and it can be used as an indicator for PQ severity.There is important clinical significance in detecting the change of serum level of PCT for estimating the condition of patients and evaluating the prognosis.
10.Expression of interleukin-34/colony stimulating factor-1R in the process of transforming growth factor -β1 inducing A549 cells transition
Jinxu ZHENG ; Qin ZHU ; Chao LIU ; Ping SONG ; Lirong DUAN
Acta Anatomica Sinica 2014;(3):393-397
Objective To investigate the expression of interleukin (IL)-34/colony stimulating factor(CSF)-1R in the process of transforming growth factor ( TGF)-β1 inducing epithelial-mesenchymal transition ( EMT) of human alveolar epithelial cells A549 cells.Methods A549 cells were cultured in vitro.CCK 8 was used to test the influence of the proliferative rate of A549 cells which were stimulated by TGF-β1 at different concentrations and time points .A549 cells were stimulated by 5μg/L TGF-β1 at 0 hour, the 12th hour, the 24th hour, and the 48th hour.Western blotting was adopted to detect changes of the following proteins: α-smooth muscle actin (α-SMA ) , E-cadherin ( E-Cad ) , matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1(TIMP-1). Real-time PCR was adopted to detect changes of the following genes: IL-34 mRNA, CSF-1R mRNA, MMP-2 mRNA and MMP-9 mRNA.Results TGF-β1 had no significant influence in the proliferation of A 549 cells compared with the control group(P>0.05).TGF-β1(5μg/L)stimulated A549 cells at different time point (0 hour, 12, 24, 48 hours), compared with the control group .The epithelial phenotype E-Cad protein was gradually down-regulated ( P <0.01 ) , while the mesenchymal phenotype α-SMA protein was gradually up-regulated ( P <0.01 ) and the protein of MMP-2 increased gradually (P<0.01).The protein of MMP-9 increased firstly and then was reduced (P<0.01),the peak was at the 24th hour.The protein of TIMP-1 was firstly transiently increased and then reduced (P<0.01), the minimum was at the 48th hour.Compared with the control group , the gene of IL-34 mRNA increased gradually (P<0.01), and the genes of CSF-1R mRNA, MMP-2 mRNA and MMP-9 mRNA increased firstly and then decreased ( P <0.01), which were peaked respectively at the 24th hour, the 24th hour, the 12th hour, respectively.Conclusion In the process of TGF-β1 inducing A549 cells transition,there is accompanied with the expression of IL-34/CSF-1R.