[Objective] To study the mechanisms and effects of panax notoginsenosides(PNS)on myocardial fibrosis in chronic viral myocarditic(VMC)mice.[Methods]The model mice with VMC at chronic status were established by repetitive infection of Coxsckievirus B3m(CVB3m).Heart changes of myocardium and collagen hyperplasia were observed by HE and VG stain.Collagen volume fraction(CVF)and perivascular circuferential area(PVCA)were examined by pathological examination with computed processing.Myocardium TGF-?1 was detected by immunohistochemical method and mRNA expression of TGF-?1 was analyzed by Reverse transcription polymerase chain reaction(RT-PCR)method.[Results]Marked myocardial fibrosis was observed in chronic VMC model group.Compared with blank group,the index of CVF and PVCA was increased significantly(P

Because of the rapid action and high bioavailability, traditional Chinese medicine injections (TCMIs) had been widely used in clinical critical field. In recent years, with the increasing reports of clinical adverse reaction, more and more attention was paid to them, and acute allergic reaction was the main adverse reaction. Acute allergic reaction included type-I anaphylaxis reaction and anaphylactoid reaction, the latter had been found in a variety of TCMIs and accounted for 77% of adverse reaction. But the mechanism of anaphylactoid reaction was not completely understood, the standard animal model for TCMIs was not established, and the technical guidance for anaphylactoid reaction was not formulated. Thus the three aspects included mechanism, evaluation index and evaluation methods of TCMIs for anaphylactoid were reviewed. Five ways including direct stimulating pathway, complement pathway, coagulation pathway, kallikrein-kinin pathway and acute allergic pathway were the main mechanism of anaphylactoid reaction; whole animal model and cell model were the main evaluation methods; the occurrence index and effect index were reviewed for the evaluation index analysis.
Anaphylaxis ; chemically induced ; Animals ; Drug Hypersensitivity ; diagnosis ; etiology ; Humans ; Injections ; Medicine, Chinese Traditional ; adverse effects

[Objective]To study the effect of TGF?1 on the expression of fibronectin and ?1 integrin receptor in chronic viral myocarditis of myocardial fibrosis induced by repeated virus infection and the interventional effect of QingXin.[Methods] Establish mice model of myocardial fibrosis in chronic stage of viralmyocarditis,then divide the mice model into model group,captopril group,QingXinⅡ high dose,medium dose and low dose groups.The normal group and model group are given saline through stomach perfusing,as well as the treated group are given respectively captopril and QingXinⅡ high,medium and low dose in the same way.After the treatment,hearts are collected,CVB3-RNA are detected by RT-PCR,TGF?1 and FN are detected by immunohistochemical technique,integrin?1 are analyzed by immunofluorescence staining-confocal laser scanning.[Results] After having done the model,TGF?1,integrin?1,FN has increased obviously;virus can be detected in themyocardium by RT-PCR;after the intervention of QingXinⅡ,TGF?1,integrin?1,FN has decreased,virus has reduced in treated group.[Conclusion] Virus,TGF?1,integrin?1 and FN have important effects on the formation ofmyocardial fibrosis;QingXinⅡ has the function of antivirus and anti-myocardial fibrosis.

Objective To construct a mammalian vector encoding angiostatin kringle 5 (K5) under the control of ?-fetoprotein (AFP) enhancer and albumin promoter, and to observe the expression of angiostatin by introducting angiostatin gene into hepatocellular carcinoma cells through gene transfection. Methods Angiostatin cDNA was amplified from normal human eukaryotic cells by using RT-PCR. Meanwhile, AFP enhancer and albumin promoter sequences were directed cloned and were inserted into vector pcDNA3.1. The recombinant vector of pcDNA3.1-AFAB-angiostatin K5-His was constructed, which contained the angiostatin K5 cDNA sequence that was under the control of the AFP enhancer and promoter. Angiostatin K5 cDNA was introduced into human AFP positive hepatocellular carcinoma cell lines with the transfected cultured cells that were mediated with Lipofectamine 2000. The expression of angiostatin K5 was analyzed by Western blot and the protein was dectected with anti-His antibody. Results The 500-base pair of angiostatin K5 was in accordance with the expected sequence and the recombinant vector of pcDNA3.1-AFAB-angiostatin K5-His was also confirmed as the anticipated sequence. The expression of angiostatin K5 in AFP positive hepatocellular carcinoma cells was detected both by SDS-PAGE and Western blot. Conclusion Efficient construction and expression of angiostatin K5 to AFP positive cells make it possible for antiangiogenesis therapy of human hepatocellular carcinomas, which may provide a promising approach.

The two pabAB genes encoding aminodeoxychorismate synthase(ADC synthase)from a L-serine producing strain Corynebacterium glutamicum SYPS-062 and model strain Corynebacterium glutamicum ATCC 13032 were ampilified by PCR.The result of nucleotide sequence analysis showed that both pabAB fragments were 1863bp,encoding 620 amino acids.16 bases differences that resulted in the changes of 7 amino acids were found in the pabAB of SYPS-062.The two pabAB were inserted into pET-28a to yield the recombinant expression vector pET-28a-pabAB and then transfromed into BL21(DE3).Upon IPTG induction,soluble ADC synthase was over-produced by E.coli BL21(DE3)harboring the expression construct.Recombinant ADC synthase purified by Ni-NTA affinity chromatography showed a single band about 67kDa on SDS-PAGE gel,and activity of aminodeoxychorismate synthase analysis show that the enzyme specific activity of SYPS-062 is 46.6% lower than ATCC 13032.

Fatty Acids ; metabolism ; Humans ; Myocardial Ischemia ; diagnostic imaging ; metabolism ; Tomography, Emission-Computed, Single-Photon

Background and purpose:Ca2+plays a very important role in the maintenance of cell biological functions. The storage, release and uptake capacity of Ca2+ is controlled by endoplasmic reticulum (ER). Ca2+ homeo-stasis is essential for cellular energy metabolism and proper protein folding. This study aimed to investigate the effect of cytoplasmic Ca2+ on cisplatin induced ER stress-mediated autophagy in ovarian carcinoma SKOV3 and its underlying mechanism.Methods:The ovarian cancer SKOV3 was used as a study object. The experiment consisted of three parts:① To explore the possible relationship between cisplatin-induced ER stress and autophagy, SKOV3 cells were treated with cisplatin for 0, 6, 12 and 24 h, respectively;② To explore the possible relationship between ER stress induced Ca2+ effux and autophagy, SKOV3 cells were treated with cisplatin for 0, 9 and 12 h, respectively, and TG was used as a positive control;③ To explore the effects of blocking calcium effux on autophagy, SKOV3 cells were divided into control group, cisplatin group, TG group, BAPTA-AM group, cisplatin combined with BAPTA-AM group and TG com-bined with BAPTA-AM group. Western blot was used to detect the protein levels of GRP78 and LC3. Fluo-4 calcium lfuorescent probe was used to examine cytoplasmic Ca2+ levels. Confocal microscopy was used to detect LC3 level by immunolfurescence staining.Results:Compared to control group (0.679±0.011), GRP78 was signiifcantly accumulated at 6, 12 and 24 h after cisplatin treatment and reached the maximum value at 6 h (1.393±0.004,P=0.000). Similarly, compared to control group (0.038±0.000), LC3 puncta were clearly seen after cisplatin treatment and reached the maxi-mum value at 12 h (0.072±0.002,P=0.000). Using confocal microscopy, we found that cisplatin and TG increased LC3 punctate accumulation and cytoplasmic Ca2+ levels in a time-dependent manner. Immunolfuorescent method showed that treatment with cisplatin combined with BAPTA-AM or TG combined with BAPTA-AM increased LC3 punctate accumulation induced by cisplatin or TG. The results of Western blot showed that cisplatin combined with BAPTA-AM (0.071±0.001) or TG combined with BAPTA-AM (0.065±0.001) signiifcantly increased LC3Ⅱ/LC3Ⅰ ratio induced by cisplatin (0.039±0.000,P=0.000) or TG (0.035±0.001,P=0.000).Conclusion:Cisplatin induces intracellular ER stress and autophagy in SKOV3 cells, accompanied by increased cytoplasmic Ca2+ levels. Chelating cytoplasmic Ca2+enhanc-es cisplatin-induced autophagy.

Objective To observe the effect of Astragalus injection on the expressions of inflammatory cytokines in human primary macrophages stimulated by lipoteichoic acid (LTA) and lipopolysaccharide (LPS),and investigate its effects on inflammatory reactions of Gram-positive (G+) and Gram-negative (G-) bacteria sepsis and its mechanisms.Methods Percoll density gradient centrifugation was used to isolate the human peripheral blood mononuclear cells,then they were purified by immune Anti-Biotin Microbeads with magnetic character and under the induction of recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF),the cells were cultivated for 12 days in vitro,eventually the human monocyte-derived macrophage was formed.The cultured human macrophages were inoculated in 96-well plates (each group 3 wells) and 6-well plates (each group 3 wells).The cells were divided into control group (200 μL DMEM added in each well),LTA 1 mg/L group,LPS 0.1 mg/L group and low astragalus injection (0.1 mg/L) and high astragalus injection (0.2 mg/L) dose groups.After the incubator plates were put in an incubator for 24 hours,the protein content of IL-8 and IL-10 in supernatant were detected by enzymelinked immunosorbent assay (ELISA),and the mRNA expression levels of IL-8 and IL-10 were detected by real time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR).Results LTA and LPS all can obviously up-regulate the expression levels of pro-inflammatory factor IL-8 and anti-inflammatory factor IL-10 of macrophage.The expressions of IL-8 and IL-10 protein and mRNA in LTA group and LPS group were significantly higher than those in control group after cuhure for 8 hours and 24 hours,the degrees of increment were more significantly at 24 hours [LTA stimute group:IL-8 protein (ng/L,× 103):41.57± 1.90 vs.1.58 ±0.24,IL-8 mRNA (A value):21.49±8.05 vs.1.00±0.16;IL-10 protein (ng/L):5.90±3.02 vs.2.91 ± 1.54,IL-10 mRNA (A value):1.35±0.34 vs.0.95±0.14;LPS stimute group:IL-8 protein (ng/L,× 103):345.00±22.80 vs.5.60±0.31,IL-8 mRNA (A value):29.84 ± 8.93 vs.1.00 ± 0.16,IL-10 protein (ng/L):122.37 ± 39.26 vs.44.79 ± 3.67,IL-10 mRNA (A value):7.38 ± 1.58 vs.1.35 ± 0.34,all P ＜ 0.05].The Astragalus injection could regulate LTA and LPS to stimulate the macrophage to decrease the expression levels of pro-inflammatory factor IL-8 protein and mRNA and increase the expression levels of anti-inflammatory factor IL-10 protein and mRNA in the macrophage;the changes of regulatory effect in the 24 hour-culture of Astragalus injection high dose group was the most significant [LTA stimute group:IL-8 protein (ng/L,×103):22.63±1.91 vs.41.57±1.90,IL-8 mRNA (A value):12.10±1.93 vs.21.49±8.05,IL-10 protein (ng/L):14.03±2.22 vs.5.90±3.02,IL-10 mRNA (A value):10.37±6.08 vs.1.35±0.34;LPS stimute group:IL-8 protein (ng/L,× 103):167.75 ± 19.90 vs.345.01 ±22.80,IL-8 mRNA (A value):15.61 ± 3.63 vs.29.84±8.93;IL-10 protein (ng/L):243.22±14.41 vs.122.37±39.26,IL-10 mRNA (A value):16.14±4.10 vs.7.38± 1.58,all P ＜ 0.05].Conclusions In the process of inflammatory response,the pro-inflammatory and anti-inflammatory factors co-exist simultaneously.Astragalus injection can inhibit the expression levels of pro-inflammatory factor gene and protein in the inflammatory response of G+ and G-bacteria sepsis and in the mean time,it can promote the expression levels of anti-inflammatory factor gene and protein,thus the immune mechanism of sepsis is affected,achieving the balance between pro-inflammation and anti-inflammation.

Objective To investigate the changes of cerebral regional glucose metabolism and regional homogeneity (ReHo) and their relation in patients with major depression disorder (MDD) using 18F-FDG PET/CT and functional MRI (fMRI).Methods A total of 18 MDD patients (6 males,12 females,age:(33.00±7.59) years) and 17 healthy controls (6 males,11 females,age:(34.59±8.96) years) underwent 18F-FDG PET/CT and fMRI.The changes of glucose metabolism on PET and ReHo on fMRI were analyzed individually by SPM and ReHo fMRI 1.0 software.Pearson correlation analysis was used.Results Compared with the glucose metabolism of control subjects,those of MDD patients decreased in the bilateral superior,middle and inferior frontal gyri,bilateral superior and middle temporal gyri,bilateral anterior cingulate cortices,bilateral putamina and caudate nuclei and the left pallidum.Meanwhile the glucose metabolism increased in the bilateral hippocampi and the left thalamus.The ReHo in MDD patients decreased in bilateral superior and middle frontal gyri,left pallidum,bilateral putamina,left anterior cingulate cortex,whereas increased ReHo was found in right hippocampus and right thalamus.The SUV of bilateral superior,middle and inferior frontal gyri,bilateral superior and middle temporal gyri,bilateral putamina,left caudate,left pallidum,left anterior cingulate cortex,bilateral hippocampi and bilateral thalami were correlated with ReHo (r =0.51-0.83,all P＜0.05).However,no correlation was found between the SUV and ReHo in right caudate and anterior cingulate cortex (r=0.41,0.37; both P＞0.05).Conclusion There may be relative characteristic models of abnormal cerebral metabolism and cerebral dysfunction impairment in MDD patients,and the changes of cerebral regional glucose metabolism may be correlated with the changes of ReHo.

Objective: To establish a risk prediction model and scoring system in patients with side branch (SB) occlusion during coronary bifurcation intervention. Methods: A total of 7007 consecutive patients who received percutanenous coronary intervention (PCI) in our hospital from 2012-02 to 2012-07 were recruited and 1545 patients (with 1601 bifurcation lesions) treated by single stent technique or main vessel stenting ifrst strategy were selected for our study. According to weather SB occlusion occurred during operation, the lesions were divided into 2 groups: Non-SB occlusion group,n=1431 and SB occlusion group,n=114. The data set of the ifrst 1200/1601 lesions by time sequence, was used for establishing the risk model and scoring system, the data set of rest 401 lesions was used for model validation. Results: The modeling data set presented that the relationship between pre-operative main vessel plaque and the position of branch vessel, the main blood vessel pre-stenting TIMI grade, the stenosis degree of pre-operative bifurcation nucleus, the angle of pre-operative bifurcation and the ratio of pre-senting stenosis degree of branch diameter and pre-operative main vessel to branch vessel diameter were the independent risk factors for branch occlusion. The risk model ROC=0.80, 95% CI 0.75-0.85, Hosmer-Lemeshow HLP=1.00; the scoring system ROC=0.76, 95% CI 0.71-0.82, HLP=0.12. The validation data set ROC=0.81, 95% CI 0.73-0.89, HLP=0.77; the scoring system ROC=0.77, 95% CI 0.69-0.86, HLP=0.58. The quartile integration of both data sets indicated that the patients with the integration score ≥ 10 had the higher risk for SB occlusion than those with integration score < 10 during the operation,P<0.001. Conclusion: Our research developed a simple and user-friendly system, it may distinguish the patients with high risk of SB occlusion during bifurcation intervention by quantitative stratiifcation of coronary angiographic imaging.