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Objective To explore the molecular mechanism underlying the anti-apoptotic activity of pORF5 plasmid protein of Chlamydia trachomatis,so as to provide an experimental basis for further clarifying the pathogenesis of Chlamydia trachomatis.Methods HeLa cells were divided into two groups:carbonyl cyanide m-chlorophenyl hydrazone (CCCP,an apoptosis inducer) group was stimulated by CCCP for 30 minutes,and pORF5 + CCCP group was pretreated with pORF5 plasmid protein for 18 hours followed by CCCP for 30 minutes.Then,Western blot analysis was performed to determine the expression of apoptosisrelated proteins Bcl-2,Bax and caspase-3,JC-1 fluorescent probe was used to detect changes in the mitochondrial membrane potential in HeLa cells,and cytochrome c release from mitochondria was analyzed by indirect immunofluorescence assay.To analyze whether high-mobility group box 1 (HMGB1) protein participated in the anti-apoptotic role of pORF5 plasmid protein,HMGB 1 shRNA and control RNA were separately transfected into the HeLa cells,which were then stimulated by pORF5 plasmid protein and CCCP.Then,the protein expression of Bcl-2,Bax,activated caspase-3 was determined,and cytochrome c release was analyzed.Data were compared between two groups by using paired t test.Results pORF5 plasmid protein could antagonize the CCCP-induced decrease of mitochondrial membrane potential,and the red/green fluorescence intensity ratio was significantly lower in the CCCP group (0.4 ± 0.1) than in the pORF5 + CCCP group (1.7 ± 0.3;t =6.95,P ＜ 0.01).The protein expression of Bcl-2 in the HeLa cells in the pORF5 + CCCP group was 5.3 ± 0.6 times more than that in the CCCP group (t =8.62,P ＜ 0.01),while the protein expression of Bax and activated caspase-3 in the pORF5 + CCCP group significantly decreased by 79％ ± 10％ (t =9.23,P ＜ 0.01) and 75％ ± 8％ (t =4.26,P ＜ 0.05) respectively compared with the CCCP group.Compared with the control RNA transfection group,the HMGB1 shRNA transfection group showed significantly decreased mitochondrial membrane potential in the HeLa cells (t =11.23,P ＜ 0.01),increased cytochrome c release,decreased Bcl-2 expresson (t =7.19,P ＜ 0.05) and increased Bax expression (t =13.06,P ＜ 0.01) after stimulation with pORF5 and CCCP.Conclusion Chlamydia trachomatis plasmid protein pORF5 plays an anti-apoptosis role by blocking the mitochondrial apoptotic pathway through HMGB1 protein.
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Objective:To study the relationship between apoptosis and the pORF5 protein of Chlamydia trachomatis,and further to explore its molecular mechanisms,which could lay a foundation for chlamydial pathogenic mechanisms.Methods:pGEX-6p/pORF5 recombinant expression vector was transformed to XL1-blue E.Coli to express GST-pORF5 fusion protein,and GST-pORF5 fusion protein was purified with Glutathione Sepharose 4B Beads,and cleaved to get pORF5 protein without GST tag by PreScission protease.The pORF5 protein was used to stimulate HeLa cells at different concentrations,then Western blot was used to evaluate the ex-pression of Bax,Bcl-2 and phosphorylation of PI3K/Akt at different time points,Hoechst staining and Flow cytometry were applied to measure the apoptosis of HeLa cells.Before treated with pORF5 protein for 24 h,HeLa cells were pretreated with PI3K inhibitor LY294002 for 1 h,the expression of Bax,Bcl-2 and the phosphorylation of Akt were evaluated by Western blot,apoptosis rates were also determined.Results:The pORF5 protein changed the expression of Bax and Bcl-2 in dose-and time-dependent manners,pORF5 increased the expression of Bcl-2 and decreased the expression of Bax at the concentration of 10 μg/ml,and there was obvious change at concentration of 15 μg/ml for 24 h.The apoptosis rates of pORF5 treated group were reduced by 27.3% and 8.4% respectively when compared with TNF-α treated group(P<0.01) and untreated group (P<0.05).Akt was phosphorylated after stimulated with pORF5 protein for 15 min,and reached its peak at 30 min.PI3K/Akt inhibitor led to the decrease of the expression of Bcl-2 and phosphorylation of Akt and increase of the expression of Bax,furthermore,PI3K/Akt inhibitor reversed pORF5-mediated anti-apoptosis, the apoptosis rate in LY294002 treated group was increased by 13.0%,when compared with the control group(P<0.01).Conclusion:pORF5 protein could inhibit apoptosis through activating PI3K/Akt signal pathway by induction of Bcl-2 and suppression of Bax.
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Objective To analyze the effectiveness and safety of chest pain centers in the management of patients with acute chest pain.Methods The clinical data of 315 patients with acute chest pain who presented to the Affiliated Hospital of Chengdu University of Traditional Chinese Medicine from January 2012 to December 2016 were retrospectively analyzed.The chest pain center of our hospital was established in December 2014.A total of 123 patients with acute chest pain who were treated before the establishment of the chest pain were included as the control group.From December 2014 to December 2016,192 patients with chest pain were admitted and included as the observation group.The percentages of acute myocardial infarction and patients receiving emergency intervention(percutaneous coronary intervention,PCI),the door-to-balloon(D to B)time,average length of hospital stay and rates adverse events between the two groups were compared.Results The percentages of acute myocardial infarction among patients with acute chest pain was 75.6％in the control group and 82.3％in the observation group(P=0.027).The emergency PCI rate was 83.9％in the control group and 92.4％in the observation group(P=0.001).The door-to-balloon time was(64.12±34.76)min in the control group and(58.08±16.26)min in the observation group(P=0.025).The average length of hospital stay was(10.09±4.03)days for the control group,and(7.41±3.78)days for the observation group(P=0.025).There was no statistical difference in the incidence of recurrent myocardial infarction between the 2 groups(P＞0.05).The rates of sudden cardiac death,heart failure,cardiogenic shock and adverse events were all significantly higher in the control group(all P＜0.05).Conclusions The establishment of chest pain center provides safer and more effective treatment to patients with acute chest pain.
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Objective To study the clinical value of magnetically controlled capsule endoscopy in diseases screening. Method We retrospectively analyzed 61 cases which were evaluated by magnetically controlled capsule endoscopy from March 2015 to December 2016. The items include operating time, the divergence rate score and cleanliness score of stomach. The consistency was compared between magnetically controlled capsule endoscopy and gastric duodenal endoscopy. Results 61 upper gastrointestinal tract studies were included. The mean age was (49.4 ± 11.6) years. No capsule retention, perforation or bleeding occurred. There was 98.4% patients, which cleanliness of stomach was good. There was 68.9% patients, which filling degree of stomach was good. The concordance rate of the two tests of gastrduodenoscopy and magnetically controlled capsule endoscopy was 89.9% (80/89). The concordance rate of the two tests was 78.9% (15/19) in esophageal and cardia, 92.9% (52/56) in stomach, 92.9% (13/14) in duodenum. Conclusion Our experience shows that magnetically controlled capsule endoscopy is a safe and useful tool for the diagnosis of upper gastrointestinal tract disease. The detection rate is similar to gastrduodenoscopy.
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This study was aimed to evaluate the efficacy and safety of Y ixin Tongmai Decoction in prevention and treatment of diabetes mellitus complicated with coronary heart disease ( CHD ) after coronary stenting restenosis . Sixty cases were randomly divided into the conventional western medicine treatment plus Y ixin Tongmai Decoction group ( treatment group ) and conventional treatment of western medicine group ( control group ) . Observation was given on the in-stent restenosis before treatment and one-year after treatment . And the traditional Chinese medicine ( TCM ) Syndrome Scale , changes of single symptom in TCM syndrome , effect of Y ixin Tongmai Decoction on blood glucose and other adverse reactions were also observed before and one-year after treatment . The results showed that in the curative effect evaluation , the restenosis rate of treatment group was 6 . 90%, and the restenosis rate of control group was 17 . 24%. There was significant difference be-tween two groups ( P < 0 . 05 ) . According to the standard of TCM syndrome curative effect , the total effective-ness in the treatment group was 86 . 21%, and that of the control group was 31 . 03%. And there was signifi-cant difference between two groups ( P < 0 . 05 ) . In the safety evaluation , there were no obvious abnormalities during safety detection in the clinical study of two groups. There was no influence on blood sugar levels. And no drug related adverse reactions occurred . It was concluded that Y ixin Tongmai Decoction can reduce coro-nary stent restenosis rate in diabetes complicated with CHD (chest pain) with the syndrome of qi deficiency and blood stasis , phlegm-turbid stasis . It can significantly improve the TCM syndrome and single symptom of patients with good safety .
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In recent years , along with the continuous accumulation of clinical experiences and the improvement of surgical devices, the percutaneous coronary intervention (PCI) has become a perfect treatment method and been widely used in the clinic . Hence , the coronary heart disease ( CHD ) patients have received better treat-ment results. However, the in-stent restenosis is still a big problem after PCI. The combination of Chinese and modern medicine in the prevention of restenosis has become a focused hotspot . In order to further under-stand this issue , this article discussed the Chinese and modern medicine mechanism in order to provide more thoughts for clinical practice .
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Two effective flavobacterium strains FCN1 and FCN2 were isolated from the sludge which had been contaminated by coke-plant waste water for a long time.Before isolation,the naphthalene was used to cultivate such sludge for seven weeks as the sole carbon source and its concentration adding to the cultivating system was increased up to 50mg/L gradually.The polycyclic aromatic hydrocarbon degrading characteristics of our isolates themselves and with inorganic ions was studied respectively.The testing results show that our two isolates can transform and degrade anthracene,phenanthrene and pyrene.After 10 hours reaction,the removing rates of anthracene,phenanthrene and pyrene by FCN1 were 84%,69%,80% and by FCN2 were 76%,40%,71% respectively.After 106 hours,the removing rates of TOC caused by anthracene,phenanthrene and pyrene were 70%,54%,69% by FCN1 and 63%,50%,46% by FCN2.The Fe 3+ and Mg 2+ can accelerate the degrading reaction of FCN1.