ABSTRACT:In the present study ,we aimed to identify differentially expressed proteins between induced worms (the infec‐ted mice were treated intragastrically with ED50 PZQ) and uninduced worms (control group) for clarifying the mechanism of PZQ .ED50 PZQ was used to administrate mice that were infected with S .japonicum via intragastric incubation for consecutive‐ly 30 days .Twenty‐one days later ,mice were sacrificed after treatment with 200 mg/kg PZQ for continuously five days ,and the male worms were obtained and some of them were subjected in DMEM medium with different concentrations of PZQ in vitro for 16 hours .Then the worms were washed twice and incubated in PZQ‐free medium for 72 hours .Compared with control group ,the induced worms had lesser sensitivity to PZQ .The survival rate of induced worms was 75 .6% in vitro when the con‐centration of PZQ was 112 mol/L (the concentration was 8 times of uninduced worms Lethal Concentration ) ,significantly higher than that in the uninduced worms (11 .1% ,P<0 .05) ,showing obviously tolerance .The other induced and uninduced worms were acquired and collected for 2D‐DIGE and MALDI‐TOF‐MS ,and combined with bioinformatics to analyse the func‐tion of the identified protein .Thirty differential expression proteins were confirmed between induced and uninduced worms ,in‐cluding 12 proteins up‐regulated and 18 proteins down‐regulated .These proteins respectively ascribed to cytoskeleton‐associat‐ed protein ,glucose and energy metabolism enzymes ,stress proteins ,thioredoxin peroxidase enzymes ,and other protease .Up‐or down‐regulation of these differential proteins indicated that PZQ promote or inhibit the expression of some specific genes . These findings may help to clarify the mechanism of PZQ ,simultaneously ,providing a scientific basis for exploring new vaccine candidate antigens and targets for drug therapy .

The protoplasts of Micromonospora purpurea,the high yield strains of gentammicin producer were mutagenized by diethyl sulfate(DES)and ultraviolet radiation(UV)respectively,then fused,screened by gentamicin resistance and regenerated.The average fermentation unit 2200?U/ml could be achieved by shake flask for 10 batches.The average fermentation unit 1900?U/ml could be obtained by 5L fermentor for 7 batches.The quality of the end product conformed to CP2000,BP2000 and USP26 pharmacopoeia.

This study investigated the effect of arctigenin (Arc) on the cell activation, cytokines expression, proliferation, and cell-cycle distribution of mouse T lymphocytes. Mouse lymphocytes were prepared from lymph node and treated with Phorbol-12-myristate-13-acetate (PMA)/Ionimycin (Ion) and/or Arc. CD69, CD25, cytokines, proliferation and cell cycle were assayed by flow cytometry. The results showed that, at concentrations of less than 1.00 micromol x L(-1), Arc expressed non-obvious cell damage to cultured lymphocytes, however, it could significantly down-regulate the expression of CD69 and CD25, as well as TNF-alpha, IFN-gamma, IL-2, IL-4, IL-6 and IL-10 on PMA/Ion stimulated lymphocytes. At the same time, Arc could also inhibit the proliferation of PMA/Ion-activated lymphocytes and exhibited lymphocyte G 0/G1 phase cycle arrest. These results suggest that Arc possesses significant anti-inflammatory effects that may be mediated through the regulation of cell activation, cytokines expression and cell proliferation.
Animals ; Anti-Inflammatory Agents ; isolation & purification ; pharmacology ; Antigens, CD ; metabolism ; Antigens, Differentiation, T-Lymphocyte ; metabolism ; Arctium ; chemistry ; Cell Cycle Checkpoints ; drug effects ; Cell Proliferation ; drug effects ; Cytokines ; metabolism ; Female ; Furans ; isolation & purification ; pharmacology ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-6 ; metabolism ; Ionomycin ; pharmacology ; Lectins, C-Type ; metabolism ; Lignans ; isolation & purification ; pharmacology ; Lymphocyte Activation ; drug effects ; Mice ; Mice, Inbred BALB C ; Plants, Medicinal ; chemistry ; T-Lymphocytes ; cytology ; drug effects ; immunology ; Tetradecanoylphorbol Acetate ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo evaluate the value of brain natriuretic peptide (BNP) in estimating risk stratification in patients with acute myocardial infarction (AMI) and to determine the relationship between BNP and adverse cardiac events after AMI.

METHODSThe 135 subjects were selected into the study, including 25 healthy subjects and 110 patients with a first AMI. The plasma concentrations of BNP were measured at two to four days after infarction in patients and healthy controls. Left ventricular function was evaluated by echocardiography with the parameters of left ventricular ejection function (LVEF) after 3 months. Patients were followed up at 12 months. The main outcome measures were heart failure, left remodeling, mortality and other adverse cardiac events at one year.

RESULTSPlasma BNP concentrations in patients with AMI were much higher than those in the health control people (416.7 +/- 208.0 ng/L versus 61.8 +/- 34.1 ng/L, P < 0.01). The BNP count ranged from 5 to 2500 ng/L in AMI patients. There was no association between the BNP count and mortality rate. The development of new congestive heart failure (CHF) was associated with a higher BNP count (P = 0.02). The development of any of the clinical end points (death/CHF/shock) occurred more frequently in patients with a higher BNP count (13.8% for BNP count of < 100 ng/L, 39.1% for BNP count of 100 - 200 ng/L, 43.3% for BNP count of 200 - 400 ng/L, 46.4% for BNP count of > 400 ng/L; P = 0.019). Plasma BNP concentrations remained independently associated with the development of clinical end points in multivariable model that adjusted for potential confounding variables.

CONCLUSIONThe results of the present study confirm that the elevated BNP count related to the risk stratification and prognosis in patients with AMI. Elevations in BNP count are associated with a higher incidence of new CHF and adverse clinical outcomes after AMI. It could serve as a strong predictor for the subsequent development of poor outcomes in AMI patients.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; blood ; diagnosis ; Natriuretic Peptide, Brain ; blood ; Prognosis

OBJECTIVETo study the effect of Tangshenkang Granule (TG) containing serum on renal mesangial cells' (RMCs) proliferation and TGF-β1/Smad2/3 pathway in the high glucose condition.

METHODSTwelve SD rats were randomly divided into four groups, i.e., the low dose TG group, the middle dose TG group, the high dose TG group, and the blank control group, 3 in each group. After 7-day gastrogavage via portal vein blood, rats were sacrificed and their serum samples were collected. RMCs were cultured in common rat serum and TG containing serum respectively. The proliferation of mesangial cells was determined by methly thiazolyl tetrazolium (MTT) assay to determine the optimal TG containing serum concentration. Expression levels of TGF-β1 mRNA and protein were determined by real time quantitative PCR and ELISA. Smad2/3 protein expression and phosphorylation were determined by Western blot and immunofluorescence.

RESULTSTG containing serum at different doses could inhibit high glucose induced RMC cells' proliferation, TGF-β1 over-expression and Smad2/3 phosphorylation.

CONCLUSIONTG containing serum could inhibit high glucose induced RMC cells' proliferation, and its mechanism might be possibly associated with inhibiting TGF-β1/Smad2/3 signaling pathway.

Animals ; Cell Proliferation ; Drugs, Chinese Herbal ; pharmacology ; Glucose ; Mesangial Cells ; Phosphorylation ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Serum ; Signal Transduction ; Smad2 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism

ObjectiveTo investigate the efficacy and safety of elbasvir/grazoprevir in patients with genotype 1 hepatitis C in the real world. MethodsA total of 35 patients with hepatitis C who received elbasvir/grazoprevir treatment in Guangzhou Eighth People’s Hospital, Guangdong Provincial Hospital of Traditional Chinese Medicine, and Guangdong General Hospital from August 2018 to March 2019 were enrolled, treated for 12 weeks, and then followed up for 12 weeks after drug withdrawal. The patients were observed in terms of sustained virologic response at week 12 after drug withdrawal (SVR12), biochemical response, and incidence rate of adverse events during treatment and follow-up. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between groups, and the Mann-Whitney U test was used for further comparison between two groups; the chi-square test was used for comparison of categorical data between groups. A logistic regression analysis was used to investigate the risk factors for virologic response in patients with hepatitis C. ResultsAmong the 35 patients with HCV infection, 97.1% (34/35) had genotype 1b HCV and 2.9% (1/35) had genotype 1a HCV; of all patients, 28 (80%) were non-cirrhotic patients with chronic hepatitis C and 7 (20%) had compensated liver cirrhosis. At the end of treatment, the virologic response rate of 100% (28/28) and SVR12 was 94.74% (18/19). In addition, age, sex, baseline HCV RNA load, previous treatment history, presence or absence of liver cirrhosis, renal function, and presence or absence of other diseases did not affect the treatment outcome (all P＞0.05). There were significant changes in the levels of alanine aminotransferase, aspartate aminotransferase, gamma-glutamyl transpeptidase, and albumin from baseline to the end of 12-week treatment (Z=-7.131, -6.797, -3.060, and -2.875, all P＜0.05). No patient experienced drug withdrawal during treatment. ConclusionThis study confirms that elbasvir/grazoprevir has good efficacy and safety in the treatment of hepatitis C in domestic real-world studies.

Objective To investigate the distribution features of TCM patterns and differentiation method of thyroid cancer through cluster analysis on data of TCM four examinations after surgery.Methods The data of TCM four examinations were collected from 304 patients by applying cross-section epidemiological survey,and then given quantitative classification and a database was established.The quantitative scores of each patient were given cluster analysis by using sample clustering analysis method (Q clustering) for getting the proportion of each category of four examination information.The preliminary criterion for patterns of thyroid cancer was made according to the proportion of four examinations information and expert survey of Delphi method at earlier stage,which was combined with clinical practice to determine the patterns.Results All 304 patients were clustered into 9 categories indicated by from A1 to A9.The frequency of A1 was 77 cases;A2,7;A3,1;A4,7;A5,105;A6,7;A7,87;A8,11 and A9,2.Based on clinical practice and experts' experiences,A1 and A7,A5 and A6,and A3 and A4 were merged and evaluated respectively as pattern of dual deficiency of qi and yin (164 cases,53.9％),pattern of liver depression and qi stagnation (112 cases,36.8％) and spleen-kidney yang deficiency pattern (8 cases,2.6％).A2 was evaluated as pattern of yin damaged by stasis heat (7 cases,2.3％).The proportions of four examination information of A8 and A9 were disperse and evaluated as other patterns (13 cases,4.4％).Conclusion The distribution of TCM patterns in the patients with thyroid cancer after surgery has its regularity.It is feasible to evaluate TCM patterns with cluster analysis combining with experts' experiences.

This article summarized Professor Liu Qiquan's clinical experience in treating urethral syndrome from the perspective of"regulating the five internal organs to calm the spleen and stomach".It is believed that urethral syndrome is located in the urethra,and its core pathogenesis is disharmony between the spleen and stomach,obstruction of dampness,and unfavorable water channel.Therefore,"tranquilizing the spleen and stomach"should be carried out throughout the treatment.At the same time,the occurrence of the disease is closely related to the disorder of the five internal organs.External sensation,internal injury,emotion,diet,fatigue and visceral disorders will affect the function of the five internal organs and then the disease will occur.In clinical practice,based on the idea of"regulating the five internal organs to calm the spleen and stomach",according to the symptoms and manifestations of the patients,the treatment can be guided by comprehensively using the methods of raising the clear and lowering the turbid to harmonize the stomach and dredge the drench,nourishing the heart and clearing the heart to purge heat and dredge the drench,benefiting the lung and purging heat to relieve dampness and dredge the drench,tonifying the kidney and warming the yang to dissipate qi and dredge the drench,dredging the liver and clearing the liver to promote the flow of qi and dredge the drench,which can make the five internal organs harmonious,benefit the waterways.Finally,the symptoms would be reduced and the disease would be cured.

OBJECTIVETo determine if multi-detector CT (MDCT) characterization of plaque is correlated with the classification of acute coronary syndrome (ACS).

METHODSAltogether 1900 patients were examined by MDCT from December 2007 to May 2009, of whom 95 patients fulfilled the criteria of ACS. Those patients were divided into the discrete plaque group ( n=61) and diffuse plaque group ( n=34) based on the findings in MDCT. The clinical diagnosis of ACS and CT results were analyzed, including segment stenosis score, segment involvement score, 3-vessel plaque score, left main score, calcification score, and remodeling index. The incidences of major adverse cardiac events in follow-up period were also recorded.

RESULTSThe patients of the diffuse plaque group were older than those of the discrete plaque group ( Pü0.0001). The diffuse plaque group presented more cases of hypertension, peripheral artery disease, diabetes, and heart failure than discrete plaque group (all P<0.05). All the 5 patients with ST-segment elevation myocardial infarction were found in discrete plaque group. The segment stenosis score of the discrete plaque group was lower than that of the diffuse plaque group(5.15±3.55 vs. 14.91±5.37, Pü0.001). The other four scores demonstrated significant inter-group difference as well (all P<0.05). The remodeling index of thediscrete plaque group was higher (1.12±0.16 vs.0.97±0.20, Pü0.05). Follow-up data showed that major adverse cardiac events occurred more frequently in diffuse plaque group than in discrete group (29.41% vs. 11.48%, P=0.0288).

CONCLUSIONSCharacteristics of discrete and diffuse plaques may be significantly different among different classes of ACS. The diffuse plaque may present higher risk, correlated to higher mortality. The diagnosis of discrete and diffuse plaques by MDCT would provide a new insight into the prognosis and treatment of ACS.

Acute Coronary Syndrome ; classification ; Adult ; Aged ; Coronary Artery Disease ; diagnostic imaging ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Plaque, Atherosclerotic ; diagnostic imaging ; Tomography, X-Ray Computed ; methods

OBJECTIVETo construct the eukaryotic expression plasmid containing lvgA gene flanked with CpG motifs of Legionella pneumophila for its expression in NIH3T3 cells.

METHODSlvgA gene flanked with CpG motifs of Legionella pneumophila was amplified by PCR. The PCR products was inserted into the eukaryotic expression plasmid pcDNA3.1/myc-his(+) to construct the recombinant plasmid pclvgA/CpG, which was subsequently transfected into NIH3T3 cells via lipofection. Immunofluorescence analysis was carried out to detect the transient expression of the plasmid in the cells.

RESULTSSequence analysis showed that the recombinant plasmid pclvgA/CpG contained the lvgA/CpG fragment with a length of 657 bp, encoding a protein of 27.7 Ku. Immunofluorescence analysis identified the transient expression of the recombinant plasmid pclvgA/CpG in NIH3T3 cells.

CONCLUSIONThe lvgA gene flanked with CpG motifs of Legionella pneumophila has been constructed successfully, and the transient expression of the recombinant plasmid pclvgA/CpG can be detected in NIH3T3 cells.

Animals ; Bacterial Vaccines ; genetics ; CpG Islands ; genetics ; Legionella pneumophila ; genetics ; Mice ; NIH 3T3 Cells ; Plasmids ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Transfection ; Virulence Factors ; biosynthesis ; genetics