AIM: It is tested that the suppressive effects of (-)-Epigallocatechin-3-gallate (EGCG ) on the migration, invasion and RhoA expression of human lung carcinoma 95-D cells and B16BL6 melanoma cells invasion in vivo,which will possibly help to understand the molecular mechanisms by which EGCG inhibits the invasion of tumor cells. METHODS: The inhibitory effect of EGCG on the migration of 95-D cells was tested by cell migration assay. Cell invasion was analyzed by the matrigel invasion assay. Assay of tumor metastasis in an animal model, RT-PCR and Western blot analysis of expression of RhoA was also performed. RESULTS:EGCG was effective in inhibiting the migration of 95-D cells in a dose-dependent manner. EGCG dose-dependently inhibited 95-D cells invasion in vitro and 40 μmol·L-1 EGCG exhibited 79.9% inhibition and EGCG 50 mg·kg-1·d-1 for 3 weeks inhibited B16BL6 melanoma cells invasion by 71.7% in vivo. EGCG could down-regulate the expression of RhoA. CONCLUSION: EGCG strongly inhibits metastasis of 95-D cells, and the mechanism of EGCG is possible associated with the inhibition of RhoA expression.

Objective:To develop an indirect enzyme-linked immunosorbent assay (ELISA) for semi-quantification of major allergen parvalbumin in fish.Methods:The soluble proteins were prepared from both white and dark muscles of seven species of freshwater fish and five species of marine fish.Tricine-SDS-PAGE and Western blot were performed to examine the protein patterns of fish muscle extracts.Natural parvalbumin being used to make calibration curve was purified from silver carp (Hypophthalmichthy molitrix) by ammonium sulphate fractionation,followed by ion exchange and gel filtration chromatography.The molecular mass of purified protein was estimated by Tricine-SDS-PAGE and identified by Western blot with anti-frog parvalbumin monoclonal antibody PARV-19.ELISA using PARV-19 was carried out to evaluate parvalbumin contents in white and dark muscles.Results:Tricine-SDS-PAGE revealed species-specific differences in proteins of heated extracts.Western blot confirmed that the major bands were showed in Tricine-SDS-PAGE with the molecular masses of 10-14 kD corresponded to parvalbumins recognized by PARV-19 and various numbers of isoforms of parvalbumin existed in different species of fish.There might be some differences in the parvalbumin contents and the epitope region was recognized by PARV-19 based on the differences in relative intensities of protein immunodetection.The ELISA showed that the contents of parvalbumin were 4 to 33 folds higher in the white muscle than in the dark muscle and varied greatly in different species of fish.Conclusion:These results validate that the dark muscle might be less allergenic than the white muscle due to the lower content of parvalbumins,and it is suggested that the commercial anti-parvalbumin antibody PARV-19 can be used to detect parvalbumins from the commercially important species of fish tested in this study and the method we develope succeeds to detect the major allergen in various species of fish.

China ; Counseling ; Female ; HIV Infections ; diagnosis ; Humans ; Pregnancy

Objective:To observe the efficacy and safety of mechanical thrombectomy with retrievable stent in patients with acute basilar artery occlusion (ABAO).Methods:Twenty-five patients with ABAO admitted to our hospital from December 2016 to October 2019 were chosen in our study. In patients who were eligible for intravenous thrombolysis and had onset within 4.5 h, mechanical thrombectomy with Solitaire AB stent was performed besides intravenous thrombolysis. In patients who had contraindications for intravenous thrombolysis or had onset within 4.5-24 h, mechanical thrombectomy with Solitaire AB stent or mechanical thrombectomy with Solitaire AB stent combined with balloon dilatation and stenting were performed. The prognoses of the patients were determined by modified Rankin scale (mRS) 90 d after surgery. The clinical data and efficacy of the patients were analyzed retrospectively, and the differences in clinical data and prognoses between patients with cardiogenic embolization and those with large artery atherosclerosis were compared.Results:Twenty-four patients had successful recanalization, and one failed. Follow-up for 90 d showed that 11 patients had good prognosis, 8 had poor prognosis, and 6 died. In patients with cardiogenic embolization, the occlusion sites were located in the upper segment of the basilar artery ( n=4) and the middle segment of the basilar artery ( n=10); in patients with large-artery atherosclerosis, occlusion sites were located in the middle segment of the basilar artery ( n=6) and the under segment of the basilar artery ( n=5); the distribution of basilar artery occlusion sites between patients with cardiogenic embolization and those with large artery atherosclerosis was statistically different ( P<0.05). The mRS scores 90 d after surgery showed no significant differences ( P>0.05). Conclusion:Mechanical thrombectomy with retrievable stent is safe and feasible for patients with ABAO, and the occlusion sites of the basilar artery in ABAO may indicate the etiology of occlusion.

Objective: To analyze the deaths with antiretroviral treatment among adult HIV/AIDS patients in Liangshan Yi Autonomous Prefecture from 2005 to 2015, in order to understand the epidemiological characteristics and to further reduce the mortality rate in Liangshan Prefecture. Methods: The relevant information was collected through the Management Database of Antiretroviral Treatment from the National AIDS Comprehensive Prevention Information System. Results: From 2005 to 2015, a total of 14 219 adult HIV/AIDS patients received antiretroviral treatment and 1 425 death cases were reported during the treatment. The cause of death was mainly AIDS-related diseases (58.9%), and the cumulative mortality rate was 10.02%. Gender, age, the way of infection, duration of antiretroviral therapy, clinical stage when received antiretroviral therapy, and CD₄⁺ T lymphocyte levels were factors for the mortality rate (P<0.001). The mortality increased with older age, higher initiation clinical stage and lower level of CD₄⁺ T lymphocyte. Among the death cases, 82.6% were male, 1 182 (82.9%) were married or cohabited, most aged between 30-39 years old (48.6%). At the initial point of receiving antiretroviral therapy, 49.7% of the cases with CD₄⁺T lymphocytes levels< 200/μl, 61.2% of the deaths cases were>1 000 copies/ml during the last viral load test, and 16.2% of deaths were ≥500/μl in the last CD₄⁺T lymphocyte test; 44.5% of deaths were received antiretroviral treatment within one year. Conclusion Early and timely antiretroviral therapy should be carried out. It is necessary to strengthen the propaganda of antiretroviral therapy and to improve the management quality of follow-up information of antiretroviral therapy case files, and to improve the medication compliance of patients.

OBJECTIVETo investigate the survival time and its impact factors among AIDS patients who initially received antiretroviral treatment (ART) of Liangshan Yi Autonomous Prefecture, Sichuan province.

METHODSA retrospective cohort study was conducted to analyze the information of AIDS patients over 18 years old initially received ART in Liangshan Yi Autonomous Prefecture during 2005-2013, which were downloaded from Chinese AIDS Antiretroviral Therapy DATA Fax Information System. Cox proportion hazard regression model was used to identify impact factors related survival time.

RESULTSAmong 8 310 ART AIDS patients who initially received ART, their mean age was (34.59 ± 9.10) years old, 65.50% (5 443 cases) were infected with HIV through injecting drug use, the mean time from testing HIV positive to starting ART were (24.68 ± 21.69) months. 436 cases died of AIDS related diseases, 28.67% (125 cases) of them died within the first 6 months of treatment. The cumulative survival rate of receiving ART in 1, 2, 3, 4 5 years were 97.11%, 93.41%, 90.61%, 88.81%, 86.02%, respectively. Multivariate Cox regression analysis showed the male patients receiveing ART were at a higher risk death of AIDS related diseases compared to the females (HR = 1.57, 95% CI: 1.13-2.182), the patients infected with HIV through injecting drug use were at a higher risk deathcompared to the infected through heterosexual transmission (HR = 1.64, 95% CI: 1.20-2.24), before the treatment patients with tuberculosis in recent1 year had higher death hazard as compared to those without tuberculosis (HR = 1.53, 95% CI: 1.05-2.21), in the treatment of the first 3 months of AIDS related diseases or symptoms of AIDS patients had higher death hazard as compared to those not suffer these diseases (HR = 1.80, 95% CI: 1.39-2.34). The patients with baseline CD4 (+) T lymphocytes cell counts < 50/µl (HR = 9.79, 95% CI: 6.03-15.89), 50-199/µl (HR = 3.26, 95% CI: 2.32-4.59), 200-349/µl (HR = 1.69, 95% CI: 1.22-2.34), were at a higher risk death than those with CD4 (+) T lymphocytes cell counts ≥ 350/µl.

CONCLUSIONAccumulate survival rate was higher after initial antiretroviral treatment among AIDS patients in Liangshan Yi Autonomous Prefecture, Sichuan province. AIDS patients who are males, have tuberculosis in recent year, infected HIV via route of intravenous drug use, with AIDS-related illness or symptoms in 3 months before ART, lower baseline CD4 (+) T lymphocyte count have higher risk of death.

Acquired Immunodeficiency Syndrome ; drug therapy ; mortality ; Adult ; Anti-Retroviral Agents ; therapeutic use ; Asian Continental Ancestry Group ; China ; Female ; Humans ; Lymphocyte Count ; Male ; Proportional Hazards Models ; Retrospective Studies ; Risk Factors ; Substance Abuse, Intravenous ; Survival Rate ; Tuberculosis ; complications

In this study, the crude polysaccharides was extracted from Shengfupian and purified by Sevag deproteinization. Then, the purified neutral polysaccharide fragment was obtained by the DEAE-52 cellulose chromatography column and Sephadex G-100 co-lumn. The structure of polysaccharides was characterized by ultraviolet spectroscopy, infrared spectroscopy, ion chromatography, and gel permeation chromatography. To investigate the anti-inflammatory activity of Shengfupian polysaccharides, LPS was used to induce inflammation in RAW264.7 cells. The expression of the CD86 antibody on surface of M1 cells, the function of macrophages, and the content of NO and IL-6 in the supernatant were examined. An immunodepression model of H22 tumor-bearing mice was established, and the immunomodulatory activity of Shengfupian polysaccharides was evaluated based on the tumor inhibition rate, immune organ index and function, and serum cytokine levels. Research indicated that Shengfupian polysaccharides(80 251 Da) was composed of arabinose, galactose, glucose, and fructose with molar ratio of 0.004∶0.018∶0.913∶0.065. It was smooth and lumpy under the scanning electron microscope. In the concentration range of 25-200 μg·mL~(-1), Shengfupian polysaccharides exhibited little or no toxicity to RAW264.7 cells and could inhibit the polarization of cells to the M1 type and reduce the content of NO and IL-6 in the cell supernatant. It could suppress the phagocytosis of cells at the concentration of 25 μg·mL~(-1), while enhancing the phagocytosis of RAW264.7 cells within the concentration range of 100-200 μg·mL~(-1). The 200 mg·kg~(-1) Shengfupian polysaccharides could alleviate the spleen injury caused by cyclophosphamide, increase the levels of IL-1β and IL-6, and decrease the level of TNF-α in the serum of mice. In conclusion, Shengfupian polysaccharides has anti-inflammatory effect and weak immunomodulatory effect, which may the material basis of Aconm Lateralis Radix Praeparaia for dispelling cold and relieving pain.
Animals ; Mice ; Interleukin-6/genetics* ; Cytokines/metabolism* ; Polysaccharides/chemistry* ; RAW 264.7 Cells ; Anti-Inflammatory Agents/chemistry* ; Spectrophotometry, Infrared

Background: Subclinical hypothyroidism (SCH) is the most common thyroid dysfunction, and its relationship with blood pressure (BP) has been controversial. The aim of the study was to analyze the association between SCH and newly-diagnosed hypertension. Methods: Based on data from the Thyroid disease, Iodine nutrition and Diabetes Epidemiology (TIDE) study, 49,433 euthyroid individuals and 7,719 SCH patients aged ≥18 years were enrolled. Patients with a history of hypertension or thyroid disease were excluded. SCH was determined by manufacturer reference range. Overall hypertension and stage 1 and 2 hypertension were diagnosed according to the guidelines issued by the American College of Cardiology/American Heart Association in 2017. Results: The prevalence of overall hypertension (48.7%), including stage 1 (28.9%) and 2 (19.8%) hypertension, increased significantly in SCH patients compared with euthyroid subjects. With elevated serum thyroid stimulating hormone (TSH) level, the hypertension prevalence also increased significantly from the euthyroid to different SCH subgroups, which was more profound in females or subjects aged <65 years. The age- and sex-specific regression analysis further demonstrated the same trends in the general population and in the 1:1 propensity matched population. Similarly, several BP components (i.e., systolic, diastolic, and mean arterial BP) were positively associated with TSH elevation, and regression analysis also confirmed that all BP components were closely related with SCH in female subjects aged <65 years. Conclusion The prevalence of hypertension increases for patients with SCH. SCH tends to be associated with hypertension and BP components in females younger than 65 years.

Background: Subclinical hypothyroidism (SCH) is the most common thyroid dysfunction, and its relationship with blood pressure (BP) has been controversial. The aim of the study was to analyze the association between SCH and newly-diagnosed hypertension. Methods: Based on data from the Thyroid disease, Iodine nutrition and Diabetes Epidemiology (TIDE) study, 49,433 euthyroid individuals and 7,719 SCH patients aged ≥18 years were enrolled. Patients with a history of hypertension or thyroid disease were excluded. SCH was determined by manufacturer reference range. Overall hypertension and stage 1 and 2 hypertension were diagnosed according to the guidelines issued by the American College of Cardiology/American Heart Association in 2017. Results: The prevalence of overall hypertension (48.7%), including stage 1 (28.9%) and 2 (19.8%) hypertension, increased significantly in SCH patients compared with euthyroid subjects. With elevated serum thyroid stimulating hormone (TSH) level, the hypertension prevalence also increased significantly from the euthyroid to different SCH subgroups, which was more profound in females or subjects aged <65 years. The age- and sex-specific regression analysis further demonstrated the same trends in the general population and in the 1:1 propensity matched population. Similarly, several BP components (i.e., systolic, diastolic, and mean arterial BP) were positively associated with TSH elevation, and regression analysis also confirmed that all BP components were closely related with SCH in female subjects aged <65 years. Conclusion The prevalence of hypertension increases for patients with SCH. SCH tends to be associated with hypertension and BP components in females younger than 65 years.

OBJECTIVES: Staphylococcus epidermidis (S. epidermidis) is a Gram-positive opportunistic pathogen that often causes hospital infections. With the abuse of antibiotics, the resistance of S. epidermidis gradually increases, and drug repurposing has become a research hotspot in the treating of refractory drug-resistant bacterial infections. This study aims to study the antimicrobial and antibiofilm effects of simeprevir, an antiviral hepatitis drug, on S. epidermidis in vitro. METHODS: The micro-dilution assay was used to determine the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of simeprevir against S. epidermidis. Crystal violet staining assay was used to detect the biofilm inhibitory effect of simeprevir. The antimicrobial activity of simeprevir against S. epidermidis and its biofilm were explored by SYTO9/PI fluorescent staining. The combined effect between simeprevir and gentamycin was assessed by checkerboard assay and was confirmed by time-inhibition assay. RESULTS: Simeprevir showed significant antimicrobial effects against S. epidermidis type strains and clinical isolates with the MIC and MBC at 2-16 μg/mL and 4-32 μg/mL, respectively. The antimicrobial effects of simeprevir were confirmed by SYTO9/PI staining. Simeprevir at MIC could significantly inhibit and break the biofilm on cover slides. Similarly, simeprevir also significantly inhibit the biofilm formation on the surface of urine catheters either in TSB [from (0.700±0.020) to (0.050±0.004)] (t=54.03, P<0.001), or horse serum [from (1.00±0.02) to (0.13±0.01)] (t=82.78, P<0.001). Synergistic antimicrobial effect was found between simeprevir and gentamycin against S. epidermidis with the fractional inhibitory concentration index of 0.5. CONCLUSIONS Simeprevir shows antimicrobial effect and anti-biofilm activities against S. epidermidis.
Humans ; Simeprevir ; Antiviral Agents ; Anti-Bacterial Agents/pharmacology* ; Cross Infection ; Gentamicins