For screening the potential drugs as anti-liver fibrosis candidates, we established a high- throughput drug screening cell model based on COL1A1 promoter. The activity of COL1A1 promoter and luciferase reporter gene can be elevated by TGF-β1, and inhibited by candidate drugs. We constructed a recombined plasmid with COL1A1 promoter and luciferase reporter gene pGL4.17, the activity of COL1A1 promoter was reflected by fluorescence intensity. COL1A1 promoter activity was detected by Dual-Luciferase Reporter Assay System, it came that the relative luciferase activity of COL1A1 promoter was 15.98 times higher than that of control group induced by TGF-β1, showing the recombined plasmid could be used in cell model. The recombined plasmid was transfected into human hepatic stellate cells LX2, detected the effect of potential drugs, and obtained a stable expression system through stable transfection and monoclonal cell culture. A sample which could reduce COL1A1 promoter activity signally by our cell model, decreased collagen I mRNA and protein expression detected by real-time RT-PCR and Western blotting. It indicates this novel cell model can be used in high-throughput drug screening of potential anti-liver fibrosis drugs.
Collagen Type I ; genetics ; Drug Evaluation, Preclinical ; methods ; Genes, Reporter ; Hepatic Stellate Cells ; High-Throughput Screening Assays ; Humans ; Liver Cirrhosis ; drug therapy ; Luciferases ; Plasmids ; Promoter Regions, Genetic ; RNA, Messenger ; Transfection ; Transforming Growth Factor beta1 ; pharmacology

Objective To explore the role of plasminogen activator inhibitor-1(PAI-1)in development of progressive fibrosis via the inhibition of extracellular matrix degradation,and to reveal the contributive role of PAI-1 in muscular dystrophy(MD).Methods Expression and cellular localization of PAI-1 protein were examined in frozen muscle specimens obtained via biopsy from 5 patients with duchenne muscular dystrophy(DMD),3 patients with becker muscular dystrophy(BMD),9 patients with congenital muscular dystrophy(CMD) and 4 cases with normal muscle by immunohistochemistry,double immunofluorescence and Western-blot analysis.Results PAI-1 was positive only in vascular endothelial cells of normal muscle.Both immunohistochemistry and Western-blot analysis showed that PAI-1 expression distinctly increased in most dystrophic muscles of MD than that in normal muscles.Double immunolabeling revealed that PAI-1 strongly expressed in cytoplasm and nuclei of regenerating muscle fibers,macrophages,macrophage infiltrating necrotic fibers.Some activated fibroblasts in endomysium and perimysium of DMD and CMD muscles were positive for PAI-1.Conclusions The functional consequence of overexpression of PAI-1 in dystrophic muscles is unknown but the elevated local expression of PAI-1 in diseased muscles of MD and their distinct distribution pattern provide evidence that PAI-1 participate in pathogenesis of MD.

Liver fibrosis is a pathological process of the excessive accumulation of extracellular matrix, especially collagen al (I) in liver. Ultimately, hepatic fibrosis leads to cirrhosis or hepatic failure. Liver fibrosis and early cirrhosis can be reversed, thus control of the development of liver fibrosis is very important for preventive treatment of cirrhosis and hepatic failure. This is a review of potential targets for anti-hepatic fibrosis based on plenty of publications, including TGF-β1 and integrin α(v) and so on, aimed at providing novel therapeutic targets in liver fibrosis.
Collagen ; metabolism ; Humans ; Integrin alphaV ; metabolism ; Liver ; pathology ; Liver Cirrhosis ; drug therapy ; Transforming Growth Factor beta1 ; metabolism

Acupuncture has remarkable effects of pain relieving and functional restoration on injuries of soft tissue and joint due to military training. As more and more attention has been attached to the impact of psychological states and biorhythm disorder on the fighting ability of military staff, acupuncture has found its place in treating chronic fatigue, combat stress reaction, traumatic brain injury and post-traumatic stress disorder as well as regulating circadian rhythms. The therapeutic effect of acupuncture in military training-related physical damage and psychological trauma has already been proved by numerous clinical practices and researches. It is held that using acupuncture as an alternative could not only save medical resources, but also enhance the fighting ability of the army. However, the current clinical studies is facing the problem of limited sample size. Therefore, randomized controlled trials in large scale and multiple centers should be further carried out toward military staff, so as to provide more speaking evidences to the prevention and treatment of physical and psychological diseases.
Acupuncture Therapy ; Adult ; Female ; Humans ; Male ; Middle Aged ; Military Personnel ; psychology ; Pain Management ; Stress Disorders, Post-Traumatic ; therapy ; Wounds and Injuries ; therapy ; Young Adult

Objective To investigate the correlation between microembolic signal (MES) and immune inflammation in patients with acute ischemic stroke. Methods The consecutive patients with acute ischemic stroke were enroled. According to the results of MES, they were divided into either a positive group or a negative group. The Immune inflammatory indexes, demographics, and baseline clinical data in both groups were compared. Multivariate logistic regression analysis was used to analyze the independent influencing factors of MES in acute ischemic stroke. Results A total of 237 patients were enroled, including 52 in the MES positive group and 185 in the MES negative group. There were significant differences in the levels of triglyceride (2. 130 ± 0. 933 mmol/L vs. 1. 811 ± 0. 962 mmol/L; t = 2. 126, P = 0. 035), plasma fibrinogen (2. 946 ± 0. 255 g/L vs. 2. 833 ± 0. 322 g/L; t = 2. 332, P = 0. 021 ), Lp-PLA2 level ( 288. 265 ± 27. 855 μg/L vs. 261. 652 ± 29. 961 μg/L; t = 2. 897, P = 0. 004 ), as wel as the proportions of CD4 + CD25high Treg (8. 695% ± 1. 461% vs. 9. 445% ± 1. 397% ; t = 3. 386, P = 0. 001), artery stenosis ≥70% (21. 15% vs. 5. 41% ; χ2 = 10. 592, P = 0. 001 ) and smal arterial occlusive stroke (9. 62% vs. 23. 24% ; χ2 = 4.667, P = 0. 031) between the MES positive group and the MES negative group. Multivariate logistic regression analysis showed that the increased plasma fibrinogen level (odds ratio [OR] 3. 257, 95%confidence interval [CI] 1. 124 - 9. 438; P = 0. 030), artery stenosis ≥ 70% (OR 3. 585, 95% CI 1. 394 -9. 219; P = 0. 008), and the decreased ratio of Treg (OR 3. 801, 95% CI 1. 190 - 12. 148; P = 0. 024) were the independent risk factors for positive MES, and smal arterial occlusive stroke was its independent protective factor (OR 0. 244, 95% CI 0. 072 - 0. 829; P = 0. 024). Conclusions MES may be associated with immune inflammation. The relationship between stroke and immune inflammation should be taken seriously.

Objective To investigate the dynamic changes of dengue viral loads and IgM antibody in patients with dengue fever ( DF) and to analyze their relationships with disease severity. Methods A to-tal of 1 508 serum samples were collected from 1 140 hospitalized patients including 1 050 mild cases and 90 severe cases within 10 days after the onset of DF in Guangzhou in 2014. The viral loads were determined by using fluorescence quantitative RT-PCR. ELISA was performed to measure the dengue virus ( DENV)-spe-cific IgM antibody. Results In general, the DENV viral loads in patients declined gradually from 108 copies/ml on day 1 to 103 copies/ml on day 10 after the onset of DF. The viral loads in severe cases were significantly higher than those in mild cases on days 5 to 7 (P<0. 05). The positive rates of DENV RNA in serum samples also decreased with the disease progression from 100% on day 1 to 40% on day 10. Com-pared with the mild cases, the patients with severe DF showed higher positive rates of DENV RNA on day 6 and day 8 (P<0. 05). The DENV-specific IgM antibody could be detected on day 2 and the secretion of IgM antibody increased gradually with the disease progression. The levels of IgM antibody in mild cases were sig-nificantly higher than those in severe cases (P<0. 001). The positive rates of IgM antibody in patients in-creased form 8% on day 2 to 95% on day 6. Higher positive rates of IgM antibody were detected in mild ca-ses on days 5 and 6 as compared with those in patients with severe DF (P<0. 05). Conclusion High viral load and low level of IgM antibody during the fastigium of DF were closely associated with the disease severity.

During oral glucose tolerance test(OGTT),endothelium-dependent vasodilation(EDD)at different time points in impaired glucose tolerance(IGT)group was lower than that in normal control group.EDD at 60 and 120 min in IGT + vitamin C group was higher than that in IGT group(all P＜0.05).There was a negative relationship between blood glucose level and EDD during OGTT in IGT patients.

Adult ; Aged ; Bone Nails ; Female ; Femoral Fractures ; surgery ; Femur ; surgery ; Fracture Fixation, Intramedullary ; Fractures, Comminuted ; surgery ; Humans ; Internal Fixators ; Male ; Middle Aged

The samples of sulfur-fumigated Paeoniae Alba Radix acquired both by random spot check from domestic market and self-production by the research group in the laboratory were used to evaluate the effects of sulphur fumigation on the quality of Paeoniae Alba Radix by comparing sulfur-fumigated degree and character, the content of paeoniflorin and paeoniflorin sulfurous acid ester, and changes of the fingerprint. We used methods in Chinese Pharmacopeia to evaluate the character of sulfur-fumigated Paeoniae Alba Radix and determinate the content of aulfur-fumigated paeoniflorin. LC-MS method was used to analyze paeoniflorin-converted products. HPLC fingerprint methods were established to evaluate the differences on quality by similarity. Results showed that fumigated Paeoniae Alba Radix became white and its unique fragrance disappeared, along with the production of pungent sour gas. It also had a significant effect on paeoniflorin content. As sulfur smoked degree aggravated, paeoniflorin content decreased subsequently, some of which turned into paeoniflorin sulfurous acid ester, and this change was not reversible. Fingerprint also showed obvious changes. Obviously, sulfur fumigation had severe influence on the quality of Paeoniae Alba Radix, but we can control the quality of the Paeoniae Alba Radix by testing the paeoniflorin sulfurous acid ester content.
Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; Fumigation ; methods ; Paeonia ; chemistry ; Quality Control ; Sulfur ; chemistry

OBJECTIVETo evaluate the oxidative stress in patients with colorectal cancer and to investigate the relationship between oxidative stress and colorectal cancer.

METHODSSeventy-six subjects were divided into two groups (36 colorectal cancer patients as the study group and 40 normal healthy individuals as the control group). Their protein oxidation, DNA damage, lipid peroxidation and antioxidants, vitamin C, vitamin E, glutathione (GSH), and antioxidative enzymes in serum were detected.

RESULTSThe levels of protein carbonyl and advanced oxidation protein products (AOPP) were significantly higher in the study group than in the control group (P<0.01). Serum 8-OHdG was significantly increased in the study group compared to the control group (P<0.01). However, the mean serum level of MDA and conjugated diene was lower in the study group than in the control group (P<0.01). The activity of antioxidative enzymes was significantly decreased in the study group compared to the control group (P<0.01). Serum vitamins C and E concentrations were significantly reduced in the study group compared to the control group (P<0.01).

CONCLUSIONColorectal cancer is associated with oxidative stress, and assessment of oxidative stress and given antioxidants is important for the treatment and prevention of colorectal cancer.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; metabolism ; DNA Damage ; Female ; Humans ; Lipid Peroxidation ; Male ; Middle Aged ; Oxidative Stress