Analysis of Residue and Transfer Status and Risk Assessment of Plant Growth Regulator Paclobutrazol in Shenmai Granules / 医药导报

Objective To analyze the residual status,transfer behavior,and risk of paclobutrazole in the national mar-ket of Shenmai granules.Methods GC-MS/MS determined the residual amount of paclobutrazol in Shenmai granules,and ANOVA analyzed the distribution characteristics of sample residuals.The transfer rule of paclobutrazol from Ophiopogonis Radix to Shenmai granules was investigated by simulating the production process,and chronic exposure assessment was performed using the point evaluation model.Results The established method can accurately determine the residual amount of paclobutrazol in Shenmai granules.The residual amount of paclobutrazol in 85 batches of Shenmai granules ranged from 0.001 3 to 0.015 8 mg·kg-1,and there was a statistical difference in the residual amount among different enterprise samples.The transfer rate of paclobutrazol from decoction pieces to preparations was 29.8%.The chronic risk quotient(HQc)of paclobutrazol residues in Shenmai granules was 0.000 7%,far lower than 1.Conclusion There is a general presence of paclobutrazol residues in Shenmai granules.The risk assessment results show that the normal dosage of Shenmai granules does not pose an unacceptable risk to the general population.The residual distribution characteristics and process transfer rules can provide a reference for safety risk control in production enterprises.

Latent tuberculosis infection among close contacts of positive etiology pul-monary tuberculosis in Chongqing / 中国感染控制杂志

Objective To investigate the current situation and influencing factors of latent tuberculosis infection(LTBI)among close contacts of positive etiology pulmonary tuberculosis(PTB)patients,provide basis for formula-ting intervention measures for LTBI.Methods A multi-stage stratified cluster random sampling method was used to select close contacts of positive etiology PTB patients from 39 districts and counties in Chongqing City as the study objects.Demographic information was collected by questionnaire survey and the infection of Mycobacterium tuberculosis was detected by interferon gamma release assay(IGRA).The influencing factors of LTBI were analyzed by x2 test and binary logistic regression model.Results A total of 2 591 close contacts were included,the male to female ratio was 0.69∶1,with the mean age of(35.72±16.64)years.1 058 cases of LTBI were detected,Myco-bacterium tuberculosis latent infection rate was 40.83％.Univariate analysis showed that the infection rate was dif-ferent among peoples of different age,body mass index(BMI),occupation,education level,marital status,wheth-er they had chronic disease or major surgery history,whether they lived together with the indicator case,and whether the cumulative contact time with the indicator case ≥250 hours,difference were all statistically significant(all P＜0.05);infection rate presented increased trend with the increase of age and BMI(both P＜0.001),and decreased trend with the increase of education(P＜0.05).Logistic regression analysis showed that age 45-54 years old(OR=1.951,95％CI:1.031-3.693),age 55-64 years old(OR=2.473,95％CI:1.279-4.781),other occupations(OR=0.530,95％CI:0.292-0.964),teachers(OR=0.439,95％CI:0.242-0.794),students(OR=0.445,95％CI:0.233-0.851),junior high school education or below(OR=1.412,95％CI:1.025-1.944),BMI＜18.5 kg/m2(OR=0.762,95％CI:0.586-0.991),co-living with indicator cases(OR=1.621,95％CI1.316-1.997)and cumu-lative contact time with indicator cases ≥250 hours(OR=1.292,95％CI:1.083-1.540)were the influential fac-tors for LTBI(all P＜0.05).Conclusion The close contacts with positive etiology PTB have a high latent infection rate of Mycobacterium tuberculosis,and it is necessary to pay attention to close contacts of high age,farmers,and frequent contact with patients,and take timely targeted interventions to reduce the risk of occurrence of disease.

Secondary metabolites from symbiotic fungi Talaromyces amestolkiae of Syngnathus acus / 中成药

AIM To study the secondary metabolites from symbiotic fungi Talaromyces amestolkiae of Syngnathus acus Linnaeus.METHODS The methanol extract from Talaromyces amestolkiae fermentation was isolated and purified by silica gel,Sephadex LH-20,TLC and preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Ten compounds were isolated and identified as 2,4-bis(1,1-dimethylethyl)benzeneethanol(1),aspergillumarins A(2),peniciisocoumarins H(3),2-(2-hydroxypropyl)-5-methyl-7-hydroxychromone(4),6-demethylvermistatin(5),penicimarin B(6),penicimarin C(7),8-hydroxy-6-methoxy-3-methylisocoumarin(8),polygonolide(9),ganoderpurine(10).CONCLUSION Compound 1 is a new natural product.Compounds 3-5,8-10 are isolated from this fungi for the first time.

The correlation analysis between plasma sclerostin levels and bone turnover markers and inflammation factors in hemodialysis patients / 中国医师进修杂志

Objective:To explore the correlation between plasma sclerostin (SOST) and bone turnover markers and inflammatory factors in hemodialysis patients.Methods:One hundred and eight patients admitted to Changsha Central Hospital Affiliated to Nanhua University from January 2018 to May 2019 were selected. The levels of plasma SOST at admission and at 3, 6 and 12 months of dialysis were determined by enzyme-linked immunosorbent assay. They were divided into low- SOSTgroup (56 cases) and high- SOSTgroup (52 cases) based on the mean value of SOST. The levels of serum bone turnover markers β-Ⅰ collagen carboxy-terminal peptide (β-CTX) and osteocalcin (OC), propeptide of type Ⅰ procollagen (PINP), full parathyroid hormone (iPTH), N-terminal osteocalcin (N-MID-OC), inflammatory factors interleukin-1β (IL-1β), interleukin-6 (IL-6), C-reactive protein (CRP), tumor necrosis factor-α (TNF-α) were compared between the two groups, abdominal aortic calcification (ACC) score was performed, and Pearson linear correlation analysis was used to analyze the relationship between SOST level of hemodialysis patients and bone turnover markers, inflammatory factors and ACC scores.Results:The baseline levels of β-CTX, OC, PINP, iPTH, and N-MID-OC in the low- SOST group were higher than those in the high- SOST group: (976.03 ± 205.27) ng/L vs. (781.34 ± 150.45) ng/L, (175.31 ± 50.49) ng/L vs. (125.75 ± 40.17) ng/L, (321.45 ± 82.14) μg/L vs. (259.41 ± 75.36) μg/L, (345.26 ± 102.65) ng/L vs. (198.52 ± 45.71) ng/L, (19.96 ± 5.01) μg/L vs. (17.41 ± 4.23) μg/L, the differences were statistically significant ( P<0.05). The baseline levels of IL-1β, IL-6, CRP, TNF-α and ACC scores in the low- SOST group were higher than those in the high- SOST group: (19.31 ± 6.01) ng/L vs. (15.23 ± 4.75) ng/L, (76.85 ± 20.34) ng/L vs. (57.98 ± 15.02) ng/L, (8.15 ± 2.36) mg/L vs. (7.23 ± 1.79) mg/L, (178.37 ± 55.52) ng/L vs. (157.42 ± 10.15) ng/L, (5.96 ± 1.78) scores vs. (5.11 ± 1.15) scores, the differences were statistically significant ( P<0.05). After treated for 3, 6 and 12 months, the levels of β-CTX, OC, PINP, iPTH, N-MIC-OC in hemodialysis patients were increased, the level of SOST was decreased, the levels of IL-1β, IL-6, CRP, TNF-α increased and ACC scores were increased, the differences were statistically significant ( P<0.05). The Pearson linear correlation analysis showed that SOST level and bone turnover markers β-CTX ( r = -0.465, P<0.001), OC( r = -0.498, P<0.001), PINP( r = -0.511, P<0.001), iPTH ( r = -0.396, P = 0.012), N-MID -OC ( r = -0.323, P = 0.031) and inflammatory factors IL-1β( r = -0.305, P = 0.046), IL-6( r = -0.318, P = 0.041), CRP( r = -0.327, P = 0.034) and TNF-α( r = -0.378, P = 0.024) in hemodialysis patients were negatively correlated, and negatively correlated with abdominal aortic calcification scores ( r = -0.301, P = 0.048). Conclusions:Plasma SOST level in hemodialysis patients is lower, which is negatively correlated with bone turnover markers, inflammatory factors, and calcification scores. Low SOST level can induce vascular calcification by mediating bone metabolism disorders and aggravating the body′s inflammatory response, and increase the risk of hemodialysis vascular calcification.

Nucleosides-based identification model for Fritillariae Cirrhosae Bulbus / 中国中药杂志

A high performance liquid chromatography( HPLC) method was established for the fast,and precise determination of ten nucleosides in Fritillariae Cirrhosae Bulbus and its counterfeits. Then multivariate statistical analyses,such as clustering analysis,principal component analysis( PCA),and Fisher' s linear discriminant analysis( LDA),were conducted to establish a discriminant function model for an integrated analysis. The results indicated that data acquisition time of a single sample was shortened within 16 min by the HPLC method. In the range of 5-1 000 mg·kg~(-1),the mass concentrations of all nucleosides exhibited good linear relationships with the corresponding peak areas( R2> 0. 999). The spiked recoveries were in the range of 93. 83%-108. 9% with RSDs of0. 12%-1. 3%( n = 5). The limit of quantitation( LOQ) was 0. 98-4. 13 mg·kg~(-1). As revealed by the clustering analysis,Fritillariae Cirrhosae Bulbus and the counterfeits could be discriminated into two clusters based on the content of nucleosides. Fisher's LDA could achieve this discrimination,while PCA dimension reduction failed. The accuracy of the discriminant function model established on the screened characteristic indicators reached 97. 5%. The present study proposed a new identification method of Fritillariae Cirrhosae Bulbus with one-dimensional indicators,which is simple,accurate,and reliable. It can provide a scientific basis for further optimizing the identification techniques for Fritillariae Cirrhosae Bulbus and inspiration for quality control strategy development of Chinese medicinal materials.

Commelina communis L. extract inhibits hypoxia/reoxygenation-induced apoptosis of cardiomyocytes and expression of inflammatory factors through regulating NADPH oxidase 4 (Nox4) / 中华微生物学和免疫学杂志

Objective:To investigate the effects of Commelina communis L. on hypoxia/reoxygenation (H/R)-induced apoptosis of myocardial cells and expression of inflammatory factors through regulating NADPH oxidase 4 (Nox4). Methods:H9c2 cells were subjected to H/R to establish the injury model. These cells were then treated with different concentrations of Commelina communis L. extract. Cell activity and apoptosis were measured by MTT method and flow cytometry, respectively. Superoxide dismutase (SOD), malondialdehyde (MDA) and lactate dehydrofenase (LDH) levels were detected. ELISA was performed to detect TNF-α, IL-1β and IL-6 levels. Western blot was used to detect the expression of Bcl-2-associated X protein (Bax), cysteine-containing aspartic protease 3 (Caspase-3) and Nox4. Real-time quantitative PCR (qPCR) was used to measure the expression of Nox4 at mRNA level. H9c2 cells were transfected with si-Nox4 and subjected to H/R. Changes in the activity and apoptosis of H9c2 cells, and the expression of SOD, MDA, LDH and inflammatory factors were observed after inhibiting Nox4 expression. H9c2 cells transfected with pcDNA3.1-Nox4 were treated with Commelina communis L. extract and subjected to H/R to evaluate the effects on the cell activity and apoptosis, as well as the expression of SOD, MDA, LDH and inflammatory factors. Results:The survival rate and SOD activity of H9c2 cells exposed to H/R were significantly reduced, but the apoptosis rate, the levels of Caspase-3, Bax protein, MDA, LDH TNF-α, IL-1β and IL-6, and the expression of Nox4 at both mRNA and protein levels were dramatically increased ( P<0.05). As with inhibition of Nox4 expression, Commelina communis L. extract at the concentrations of 10 μg/ml and 20 μg/ml could obviously increase the survival rate and SOD activity of H9c2 cells after H/R exposure, and reduce the apoptosis rate and the expression of Caspase-3, Bax protein, MDA, LDH TNF-α, IL-1β and IL-6, as well as Nox4 expression at mRNA and protein levels ( P<0.05). Overexpression of Nox4 reversed the effects of Commelina communis L. extract on promoting cardiomyocyte cell activity and SOD activity, inhibiting cell apoptosis, and downregulating the expression of Caspase-3, Bax protein, MDA, LDH, TNF-α, IL-1β and IL-6 in the H/R injury model. Conclusions:Commelina communis L. could protect against H/R-induced myocardial cell injury, improve cell activity, and inhibit cell apoptosis and the expression of inflammatory factors through regulating Nox4 expression.

Cytological study on aptamer selection for bladder cancer through CELL-SELEX / 医学研究生学报

ObjectiveThe methods based on bladder cancer markers which could be applied to early diagnosis and postoperative recurrence monitoring of bladder cancer were current research hotspots. This study aims to screen aptamers that specifically recognize human bladder cancer cell lines (EJ, T24, BIU87) through cell-based systematic evolution of ligand by exponential enrichment (CELL-SELEX).MethodsFor CELL-SELEX screening, bladder cancer cell lines EJ, T24, and BIU87 were used as positive control cells. HCV 29 (human normal urothelial cell line), 293T (human embryonic kidney cell line), huh7 (human hepatocellular carcinoma cell line) were used as negative control cells. PCR upstream primers were labeled with FITC, downstream primer was labeled with Biotin. ssDNA fragments collected from each round were amplified by PCR, and the amplified product was then purified using a DNA purification Kit. The biotin-streptavidin magnetic separation methods were used to isolate the PCR product to obtain secondary FITC-ssDNA for the next CELL-SELEX round. The screening process was monitored by flow cytometry. ssDNA pool with the highest binding rates to bladder cancer cell lines(EJ, T24, and BIU87) was selected to PCR amplification, product purification, molecular cloning, and sequencing. According to the sequencing results, the secondary structure of the aptamer was pre-simulated by Dnaman software. Aptamer labeled with FITC was synthesized in vitro, flow cytometry was used to detect the binding rate of the aptamer to bladder cancer cell lins (EJ, T24 and BIU87).ResultsWith the advance of the CELL-SELEX process, the binding rate of FITC-ssDNA to bladder cancer cell lins (EJ, T24, and BIU87) increased gradually. By the 15th round, the binding rate of FITC-ssDNA to EJ cells reached the highest level. The apt1 had the highest enrichment among the 15th round ssDNA pool. By the 18th round, the binding rate of FITC-ssDNA to T24 or BIU87 cells reached the highest level. The apt2 and apt3 had the highest enrichment among the 18th round ssDNA pool. DNA structure prediction showed that the secondary structure of apt1, apt2, and apt3 was mainly stem-loop structure. Flow cytometry showed that the highest binding rate was FITC-apt1 to EJ cells, FITC-apt2 to T24 cells, and FITC-apt3 to BIU87 cells, respectively. There is no significant combination between these aptamers with the negative cells.ConclusionIn this study, three kinds of aptamers with high specificity for bladder cancer cell lines were successfully screened by CELL-SELEX. The apt1 can specifically recognize EJ cells, apt2 can specifically recognize T24 cells and apt3 can specifically recognize BIU87 cells, all of which provide experimental evidence for early diagnosis and targeted therapy technology research of bladder cancer.

The application of dynamic high-frequency ultrasound in the diagnosis of Zenker diverticulum / 中国医学装备

Objective:To explore the diagnostic value of high-frequency ultrasound for Zenker diverticulum.Methods: 15 patients who were suspected as Zenker diverticulum through the diagnosis of using high frequency ultrasound were analyzed in the research. Their appearances of ultrasound were summarized, and these results were compared with barium meal at upper gastrointestinal tract and results of postoperative pathology, respectively.Results: In 15 patients, 4 cases were confirmed by adopting barium meal at upper gastrointestinal tract, and 11 cases were confirmed by adopting postoperative pathology. All of lesions in 15 cases were located on the back of left side of the thyroid gland, and there were 3 kinds of sonographic appearance. The first kind was equal echo lesion, and there were spots and schistoses without echo inside lesion, their form showed a semicircle shape. The second kind was hypoechoic, the form showed irregularity or semi cyclic annular, and the border was clear, and there were strong echogenic spots. The third kind was hyperechoic lesions, and the strong echo were movable with morphological changing after the slight pressure of search unit, and slim half ring low echo wall can be seen indistinctly around lesions.Conclusion:High-frequency ultrasonography is a convenient, rapid and non-invasive method for the diagnosis of Zenker diverticulum, and it is helpful to grasp its ultrasonogram characteristic and examination method in early detection of disease, avoiding misdiagnosis and missed diagnosis. Therefore, it has important clinical value.

HPLC fingerprint of Ermiao Pills and chemical constituents analysis by UPLC-LTQ-Orbitrap / 中草药

Objective To establish a quality assessment method for Ermiao Pills (EP) based on HPLC fingerprint and qualitatively analyze the chemical constituents. Methods The chromatographic column Shiseido C18 (250 mm × 4.6 mm, 5 μm) was used, acetonitrile-0.1% formic acid as mobile phase with gradient elution at the flow rate of 1.0 mL/min, and the detection wavelength was 330 nm. The standard chromatographic fingerprint was synthesized from chromatogram of the mixed standard herbs of Phellodendron Rupr. and Atractylodes DC, and the similarity evaluation of Ermiao Pills samples was carried out by the Similarity Evaluation System for Chromatographic Fingerprints of TCM (Chinese Pharmacopoeia Commission, version 2012A). Ultra high-performance liquid chromatography coupled with linear ion trap-Orbitrap Elite mass spectrometer (UPLC-LTQ-Orbitrap) was used to characterize the chemical constituents of Ermiao Pills. A Thermo Scientific Syncronis C18 (100 mm × 2.1 mm, 1.9 μm) column and a gradient elution of acetonitrile-0.1% formic acid were used for UPLC separation. The combination of ESI-LTQ-Orbitrap mass analyzer with a linear ion trap was applied for high resolution mass spectrometry and collision-induced dissociation (CID). Results The chromatographic fingerprints were generated with 10 common peaks. The similarity scores of 20 samples between each material batch and the reference fingerprint ranged from 0.869-0.992. Twenty-one components were identified via referring to reference components and literatures and analyzing MS data, they were neo-chlorogenic acid, magnocurarine, xanthoplanine, magnoflorine, 3-O-feruloylquinic acid, menisperine, demethyleneberberine, oxyberberine, columbamine, jatrorrhizine, berberubine, palmatine, berberine, syringic acid, caffeic acid, (E)-4-(3-hydroxyprop-1-en1yl)-2-methoxyphenol, atractylenolide II, acetosyringone, atractylenolide I, selina-4(14), 7(11)-dien-8-one, and atractylodin. Conclusion The established method of fingerprint is specific, combined with LC-MS qualitative analysis, can be used for the quality evaluation of Ermiao Pills, giving support to quality control comprehensively.

Evaluation of risk factors and prognosis on diodone-induced acute kidney injury according to ESUR and KDIGO criteria / 中南大学学报(医学版)

OBJECTIVE@#To investigate the incidence, risk factors and prognosis for contrast-induced acute kidney injury (CI-AKI) according to ESUR and KDIGO criteria in patients undergoing angiography.
@*METHODS@#We evaluated 260 patients undergoing angiography and/or intervention therapy from April 2011 to January 2012 in the Second Xiangya Hospital of Central South University. All patients received low-osmolality contrast agent (ioversol). Serum creatinine was measured before angiography or at 48 or 72 h after procedure. The multivariate logistic regression was used to analyze the risk factors of CI-AKI. The major adverse events were observed in a year of follow-up.
@*RESULTS@#Among the 260 patients, 23 experienced CI-AKI and the incidence was 8.8% according to ESUR criteria. Twelve patients experienced CI-AKI and the incidence was 4.6% according to KDIGO criteria. The multivariate logistic regression analysis showed that diabetes mellitus and dehydration were the independent risk factors for CI-AKI according to ESUR criteria; In another KDIGO criteria, chronic kidney disease (CKD), hypercholesterolemia and diabetes mellitus were the independent risk factors for CI-AKI. The prognosis study showed that the mortality of patients with CI-AKI were significantly higher than those without CI-AKI (P<0.05).
@*CONCLUSION@#The incidence of CI-AKI is associated with diagnostic criteria. Diabetes mellitus, CKD, dehydration and hypercholesterolemia were the independent risk factors for CI-AKI. CI-AKI is a relevant factor for mortality in a year after angiography and/or intervention therapy.