Serum Osteopontin, KL-6, and Syndecan-4 as Potential Biomarkers in the Diagnosis of Coal Workers' Pneumoconiosis: A Case-Control Study.

Background: Coal worker's pneumoconiosis (CWP) is a chronic occupational disease mainly caused by coal dust inhalation in miners. This study aimed to investigate the clinical value of Osteopontin (OPN), KL-6, Syndecan-4 and Gremlin-1 as serum biomarkers in CWP. Patients and Methods: We integrated reported lung tissues transcriptome data in pneumoconiosis patients with silica-exposed alveolar macrophage microarray data to identify four CWP-associated serum biomarkers. The serum concentrations of Osteopontin, Krebs von den Lungen-6 (KL-6), Syndecan-4 and Gremlin-1 were measured in 100 healthy controls (HCs), 100 dust-exposed workers (DEWs) and 200 patients of CWP. Receiver operating characteristic (ROC) curve analysis was used to determine the sensitivity, specificity, cut-off value and area under the curve (AUC) value of biomarkers. Results: The pulmonary function parameters decreased sequentially, and the serum OPN, KL-6, Syndecan-4 and Gremlin-1 concentrations were increased sequentially among the HC, DEW and CWP groups. Among all participants, multivariable analysis revealed that these four biomarkers were negatively correlated with the pulmonary function parameters (all p<0.05). Compared with HCs, patients with higher OPN, KL-6, Syndecan-4 and Gremlin-1 had higher risk for CWP. The combination of OPN, KL-6, and Syndecan-4 can improve the diagnostic sensitivity and specificity of CWP patients differentiated from HCs or DEWs. Conclusion: OPN, KL-6 and Syndecan-4 are novel biomarkers that can be used for CWP auxiliary diagnosis. The combination of three biomarkers can improve the diagnostic values of CWP.

SMAD4 rs10502913 is Significantly Associated with Chronic Obstructive Pulmonary Disease in a Chinese Han Population: A Case-Control Study.

Background: COPD is a respiratory disease caused by a combination of genetic and environmental factors. Polymorphism, as a genetic factor, can affect the susceptibility of the disease of COPD. In this study, we assessed the relationship between the polymorphisms of three genes and COPD risk in a Chinese Han population. Patients and Methods: A total of 376 patients diagnosed with COPD and 284 control subjects were enrolled in this study. Multivariate logistic regression analysis was used to analyze the association between three polymorphisms (SMAD4 rs10502913, IL-4 rs2070874, HSPA1L rs2227956) and COPD susceptibility. Results: The SMAD4 rs10502913 GG and AG genotype significantly increased COPD risk (adjusted OR = 2.235, 95% CI 1.198-4.104; adjusted OR = 2.218, 95% CI 1.204-4.151, respectively) compared with the AA genotype. In the stratification analyses, the GG genotype significantly increased the risk of COPD in subjects aged 60 and over (adjusted OR = 2.519, 95% CI 1.266-5.015) and with a smoking history of less than 30 years (p=0.009; adjusted OR = 3.751; 95% CI 1.398-10.062). This increased risk was more pronounced in the group of GOLD I and GOLD II (adjusted OR = 3.628, 95% CI 1.022-12.885; adjusted OR = 2.394, 95% CI 1.004-5.710, respectively). In addition, AG genotype was associated with an increased COPD risk in subjects aged 60 and over (adjusted OR = 2.599, 95% CI 1.304-5.176) and in smokers (p=0.021; adjusted OR = 2.269; 95% CI 1.132-4.548). This increased risk was more obvious in the group of GOLD III COPD (p=0.047; adjusted OR = 2.532; 95% CI 1.012-6.336). Conclusion: Our present study indicated that the genotype GG and AG of SMAD4 rs10502913 are associated with an increased risk of COPD in a Chinese Han population. Further validation studies with large-scale populations are needed to confirm our findings.

Associations of SMAD4 rs10502913 and NLRP3 rs1539019 Polymorphisms with Risk of Coal Workers' Pneumoconiosis Susceptibility in Chinese Han Population.

Purpose: CWP is an untreatable but preventable fibrotic lung disease caused by the chronic inhalation of coal dust. Genetic factors such as polymorphisms play an important role in the development of CWP. The present study investigated the association between the polymorphisms of SMAD4 and NLRP3 and CWP risk in a Chinese Han population. Patients and Methods: SMAD4 rs10502913 and NLRP3rs1539019 polymorphisms were examined in 292 CWP subjects and 315 coal dust-exposed controls. The genotypes were analyzed using direct sequencing. The allele and genotype proportion between the cases and controls were compared using the chi-square test. Results: The AG and GG genotypes of SMAD4 rs10502913 were not associated with altered CWP risk compared with AA genotype (adjusted OR = 1.535 and 1.426, 95% CI = 0.785-3.000 and 0.732-2.781, p = 0.210 and 0.297, respectively). Also, the NLRP3 rs1539019 heterozygous and homozygous variants CA and CC genotypes were not associated with the risk of CWP compared with the AA genotype (adjusted OR = 0.985 and 1.127, 95% CI = 0.652-1.489 and 0.713-1.782, p = 0.944 and 0.608, respectively). In addition, there was no interaction between SMAD4 rs10502913 and NLRP3 rs1539019 genotypes and smoking status on CWP risk in the stratified analyses. Conclusion: In this present study, SMAD4 rs10502913 and NLRP3 rs1539019 genotypes were not associated with altered CWP risk in the Chinese Han population. Large sample sizes and multicenter studies are needed to elucidate these results in the future.

Predictive quality control for compound liquorice tablets by the intelligent mergence fingerprint method combined with the systematic quantitative fingerprint method.

INTRODUCTION: Compound liquorice tablet (CLT) is a herbal compound preparation and is used as a classic antitussive and expectorant in China. It is composed of liquorice extract powder, opioid powder, star anise oil, camphor, and sodium benzoate. The complexity of herbal materials brings a huge challenge in producing compound preparations with stable and uniform quality consistency. OBJECTIVE: To establish a new intelligent model for predicting the quality of CLT. METHODS: The HPLC fingerprints of raw materials including liquorice extract powder, powdered opium, star anise oil, and sodium benzoate were tested and merged to generate the intelligent mergence fingerprints, whose correlation with the raw materials and the CLT samples was studied. The consistency of the intelligently merged fingerprints with the standard fingerprints was observed by using the systematic quantitative fingerprint method in order to calculate quality evaluation results. RESULTS: The intelligent mergence fingerprints covered all the main fingerprint peaks of four raw materials and had a good correlation with the CLT sample fingerprint. There were no significant quality differences either among the six intelligent mergence models obtained by combining different batches of raw materials or between the reference fingerprint of the intelligent mergence connection fingerprints (RFPIMFC ) and the theoretical standard preparation (RFPS ). CONCLUSION: The computer-aided model of intelligent mergence fingerprints could be used to predict the quality of herbal compound preparations based on raw materials. In this way, preproduction quality prediction can be realised in order to avoid low-quality medicinal materials and improve the quality consistency among different batches.

Overall control herbal medicine in best consistency.

The standard preparation (SP) based on the quantitative fingerprint was proposed to control the quality of compound licorice tablets (CPLTs) in this paper, which is a great way to control the quality consistency nowadays. Here, taking China 145 batches of CPLTs from 9 manufacturers as samples to set up High Performance Liquid Chromatography fingerprints and CPLT-SP, and employing the systematically quantified fingerprint method (SQFM) combined five markers' contents to evaluate their quality. The results showed that all samples were qualified. The quantitative analysis of multi-components by single-marker (QAMS) for the determination of five components in CPLTs was applied, and there was no striking diversity in the results compared to the standard curve method. Furthermore, the correlation between raw materials and preparation of CPLTs was studied for predicting quality intelligently. As well as the ultraviolet full fingerprint dissolution was applied to evaluate the rationality of preparation. The results demonstrated that SQFM based on SP can integrally and perfectly control the quality of HD in best consistency.

Capillary electrophoresis fingerprints combined with Linear Quantitative Profiling Method to monitor the quality consistency and predict the antioxidant activity of Alkaloids of Sophora flavescens.

Alkaloids of Sophora flavescens (ASF) has various pharmacological effects, and it is widely used in clinical practice. The aim of this research was to develop an environmentally friendly methodology that enables not only identification but also the quality consistency assessment of Alkaloids of Sophora flavescens. A background electrolyte composed of 50 mmol/L sodium tetraborate solution, 500 mmol/L boric acid and 1.2 mmol/L citric acid was used to conduct the fingerprint analysis coupled with quantitative determination of three components. Linear quantitative profiling method was used for comprehensive quality discrimination of the test samples from both qualitative and quantitative perspectives, and the 27 batches of samples were well differentiated. In addition, the fingerprint-efficacy relationship between chemical components and antioxidant activity in vitro was established using partial least squares regression model, which provided important medicinal efficacy information for quality control.

GILT restricts the cellular entry mediated by the envelope glycoproteins of SARS-CoV, Ebola virus and Lassa fever virus.

Interferons (IFNs) control viral infections by inducing expression of IFN-stimulated genes (ISGs) that restrict distinct steps of viral replication. We report herein that gamma-interferon-inducible lysosomal thiol reductase (GILT), a lysosome-associated ISG, restricts the infectious entry of selected enveloped RNA viruses. Specifically, we demonstrated that GILT was constitutively expressed in lung epithelial cells and fibroblasts and its expression could be further induced by type II interferon. While overexpression of GILT inhibited the entry mediated by envelope glycoproteins of SARS coronavirus (SARS-CoV), Ebola virus (EBOV) and Lassa fever virus (LASV), depletion of GILT enhanced the entry mediated by these viral envelope glycoproteins. Furthermore, mutations that impaired the thiol reductase activity or disrupted the N-linked glycosylation, a posttranslational modification essential for its lysosomal localization, largely compromised GILT restriction of viral entry. We also found that the induction of GILT expression reduced the level and activity of cathepsin L, which is required for the entry of these RNA viruses in lysosomes. Our data indicate that GILT is a novel antiviral ISG that specifically inhibits the entry of selected enveloped RNA viruses in lysosomes via disruption of cathepsin L metabolism and function and may play a role in immune control and pathogenesis of these viruses.

[Quality consistency evaluation of Fufanggancao tablets based on the control mode of standard preparation and quantitative fingerprint].

The control mode of standard preparation (SP) and quantitative high performance liquid chromatography (HPLC) fingerprint of Fufanggancao tablets (FFGCTs) combined with the quantification of five markers were successfully developed and applied to the precise and accurate assessment of the quality consistency of 145 FFGCTs from nine manufacturers. The profiling was determined by reversed-phase HPLC at 220 nm wavelength, where the reference fingerprint (RFP) of the FFGCTs reserved as standard preparation was established. Subsequently, the SP-RFP was considered as the evaluation standard, and a systematic quantitative fingerprint method was applied to the integrative quality discrimination of 145 batches of FFGCTs, from both qualitative and quantitative perspectives. Besides, the chromatographic systematic error of quantitative fingerprints was corrected by the double marker calibration method. The results showed that the qualities of the FFGCTs from the nine manufacturers were completely qualified. Moreover, all raw herb fingerprints and the simulated sample were determined by using the combined chromatographic conditions applied to the FFGCTs, which helped identify the source of the profiling peaks in the preparation and establish the correlation between the raw herb fingerprints and the preparation fingerprints. This eventually aided the intelligent prediction of the quality of the preparation or raw herbs and effective prevention of the inputs of inferior raw materials. In addition, we employed the ultraviolet full fingerprint dissolution method to differentiate the FFGCTs from five manufacturer dissolution, in which the rationality of the preparation process was evaluated. The method is feasible and accurate, and it can be applied to evaluate the quality and process technology consistency of FFGCTs. This paper provides a fundamental standard preparation evaluation mode and the basic operation procedure for the quality consistency assessment of traditional Chinese medicine.

Identification of Residues Controlling Restriction versus Enhancing Activities of IFITM Proteins on Entry of Human Coronaviruses.

Interferon-induced transmembrane proteins (IFITMs) are restriction factors that inhibit the infectious entry of many enveloped RNA viruses. However, we demonstrated previously that human IFITM2 and IFITM3 are essential host factors facilitating the entry of human coronavirus (HCoV) OC43. In a continuing effort to decipher the molecular mechanism underlying IFITM differential modulation of HCoV entry, we investigated the roles of structural motifs important for IFITM protein posttranslational modifications, intracellular trafficking, and oligomerization in modulating the entry of five HCoVs. We found that three distinct mutations in IFITM1 or IFITM3 converted the host restriction factors to enhance entry driven by the spike proteins of severe acute respiratory syndrome coronavirus (SARS-CoV) and/or Middle East respiratory syndrome coronavirus (MERS-CoV). First, replacement of IFITM3 tyrosine 20 with either alanine or aspartic acid to mimic unphosphorylated or phosphorylated IFITM3 reduced its activity to inhibit the entry of HCoV-NL63 and -229E but enhanced the entry of SARS-CoV and MERS-CoV. Second, replacement of IFITM3 tyrosine 99 with either alanine or aspartic acid reduced its activity to inhibit the entry of HCoV-NL63 and SARS-CoV but promoted the entry of MERS-CoV. Third, deletion of the carboxyl-terminal 12 amino acid residues from IFITM1 enhanced the entry of MERS-CoV and HCoV-OC43. These findings suggest that these residues and structural motifs of IFITM proteins are key determinants for modulating the entry of HCoVs, most likely through interaction with viral and/or host cellular components at the site of viral entry to modulate the fusion of viral envelope and cellular membranes.IMPORTANCE The differential effects of IFITM proteins on the entry of HCoVs that utilize divergent entry pathways and membrane fusion mechanisms even when using the same receptor make the HCoVs a valuable system for comparative investigation of the molecular mechanisms underlying IFITM restriction or promotion of virus entry into host cells. Identification of three distinct mutations that converted IFITM1 or IFITM3 from inhibitors to enhancers of MERS-CoV or SARS-CoV spike protein-mediated entry revealed key structural motifs or residues determining the biological activities of IFITM proteins. These findings have thus paved the way for further identification of viral and host factors that interact with those structural motifs of IFITM proteins to differentially modulate the infectious entry of HCoVs.

Evaluation of the quality consistency of powdered poppy capsule extractive by an averagely linear-quantified fingerprint method in combination with antioxidant activities and two compounds analyses.

A novel averagely linear-quantified fingerprint method was proposed and successfully applied to monitor the quality consistency of alkaloids in powdered poppy capsule extractive. Averagely linear-quantified fingerprint method provided accurate qualitative and quantitative similarities for chromatographic fingerprints of Chinese herbal medicines. The stability and operability of the averagely linear-quantified fingerprint method were verified by the parameter r. The average linear qualitative similarity SL (improved based on conventional qualitative "Similarity") was used as a qualitative criterion in the averagely linear-quantified fingerprint method, and the average linear quantitative similarity PL was introduced as a quantitative one. PL was able to identify the difference in the content of all the chemical components. In addition, PL was found to be highly correlated to the contents of two alkaloid compounds (morphine and codeine). A simple flow injection analysis was developed for the determination of antioxidant capacity in Chinese Herbal Medicines, which was based on the scavenging of 2,2-diphenyl-1-picrylhydrazyl radical by antioxidants. The fingerprint-efficacy relationship linking chromatographic fingerprints and antioxidant activities was investigated utilizing orthogonal projection to latent structures method, which provided important pharmacodynamic information for Chinese herbal medicines quality control. In summary, quantitative fingerprinting based on averagely linear-quantified fingerprint method can be applied for monitoring the quality consistency of Chinese herbal medicines, and the constructed orthogonal projection to latent structures model is particularly suitable for investigating the fingerprint-efficacy relationship.

Correction: Linear Quantitative Profiling Method Fast Monitors Alkaloids of Sophora Flavescens that was Verified by Tri-marker Analyses.

[This corrects the article DOI: 10.1371/journal.pone.0161146.].

Linear Quantitative Profiling Method Fast Monitors Alkaloids of Sophora Flavescens That Was Verified by Tri-Marker Analyses.

The present study demonstrated the use of the Linear Quantitative Profiling Method (LQPM) to evaluate the quality of Alkaloids of Sophora flavescens (ASF) based on chromatographic fingerprints in an accurate, economical and fast way. Both linear qualitative and quantitative similarities were calculated in order to monitor the consistency of the samples. The results indicate that the linear qualitative similarity (LQLS) is not sufficiently discriminating due to the predominant presence of three alkaloid compounds (matrine, sophoridine and oxymatrine) in the test samples; however, the linear quantitative similarity (LQTS) was shown to be able to obviously identify the samples based on the difference in the quantitative content of all the chemical components. In addition, the fingerprint analysis was also supported by the quantitative analysis of three marker compounds. The LQTS was found to be highly correlated to the contents of the marker compounds, indicating that quantitative analysis of the marker compounds may be substituted with the LQPM based on the chromatographic fingerprints for the purpose of quantifying all chemicals of a complex sample system. Furthermore, once reference fingerprint (RFP) developed from a standard preparation in an immediate detection way and the composition similarities calculated out, LQPM could employ the classical mathematical model to effectively quantify the multiple components of ASF samples without any chemical standard.

Holistic Evaluation of Quality Consistency of Ixeris sonchifolia (Bunge) Hance Injectables by Quantitative Fingerprinting in Combination with Antioxidant Activity and Chemometric Methods.

A widely used herbal medicine, Ixeris sonchifolia (Bge.) Hance Injectable (ISHI) was investigated for quality consistency. Characteristic fingerprints of 23 batches of the ISHI samples were generated at five wavelengths and evaluated by the systematic quantitative fingerprint method (SQFM) as well as simultaneous analysis of the content of seven marker compounds. Chemometric methods, i.e., support vector machine (SVM) and principal component analysis (PCA) were performed to assist in fingerprint evaluation of the ISHI samples. Qualitative classification of the ISHI samples by SVM was consistent with PCA, and in agreement with the quantitative evaluation by SQFM. In addition, the antioxidant activities of the ISHI samples were determined by both the off-line and on-line DPPH (2, 2-diphenyl-1-picryldrazyl) radical scavenging assays. A fingerprint-efficacy relationship linking the chemical components and in vitro antioxidant activity was established and validated using the partial least squares (PLS) and orthogonal projection to latent structures (OPLS) models; and the online DPPH assay further revealed those components that had position contribution to the total antioxidant activity. Therefore, the combined use of the chemometric methods, quantitative fingerprint evaluation by SQFM, and multiple marker compound analysis in conjunction with the assay of antioxidant activity provides a powerful and holistic approach to evaluate quality consistency of herbal medicines and their preparations.

Quickly quantifying the dissolution fingerprints of compound Danshen dropping pill by HPLC.

To firstly accurately overall determine the all components of Compound Danshen Dropping Pills (CDDP) and assess how about the authentic quality is for each bath of samples through a quantified fingerprint that only can qualitatively identified which kind of a herbal or Traditional Chinese Medicine may be. The systematically quantified fingerprint method (SQFM) was employed to identify that the qualities of S4 and S6 were belong to best (grade 1), those of S3, S5, S7, S8 and S9 belong to better (grade 2), those of S2 and S10 belong to good (grade 3), and only S1 falling to fine (grade 4) where those were all with 19 fingerprint peaks. On the basis, the important HPLC dissolution fingerprints (DFPs) of CDDPs were meticulously established to investigate the dynamic process in water, 0.1 mol/L hydrogen chloride (HCl) and 0.5% sodium dodecyl sulphate (SDS), respectively, in which SQFM executed the excellently quantified analyses for the DFPs. During the evaluation, the eighth time points of DFPs were selected as the standards for all dissolving to calculate how high are the macro quantitative similarity ( P m ) of DFPs for the other different time points. Finally in terms of Weibull function, lnln [1/(1- P m )]=-0.5951 x (2)+3.8146 x -4.5195 ( r =0.9974), lnln [1/(1- P m )]=-0.5555 x (2)+3.5814 x -4.6126 ( r =0.9972) were acquired in water and in 0.1 mol/L HCl solution respectively, but without in the SDS solution. In order to specially monitor the dissolution dynamic changes of the important fingerprints to guide the preparation technology and improve their bioavailability, We investigated the four strong peaks of CDDP-DFP and calculated out the fitting equations as follows, lnln [1/(1- A i / A max )]= y , the first one (No.6, Protocatechualdehyde), y =-0.5603 x (2)+3.4516 x -3.8974 ( r =0.9988); the second one (No.19, Salvianolic acid B), y =-1.7127 x (2) +9.6655x-11.947 ( r =0.9897); the third one (No.4, Danshensu), y =-0.4239 x (2)+3.0436 x -3.6276 ( r =0.9985); the forth one (No.17, unkown), y =-0.5019 x (2)+3.3706 x -4.1053 ( r =0.9979), which indicated Salvianolic acid B with a prominent discrepancy from others. The quantified HPLC fingerprint can perfectly both quantitatively reflected the authentic quality of CDDPs and accurately overall quantitatively measure the dissolution dynamic varieties in vitro to provide the important information for bioavailability, in which water is proven to be the best dissolution medium for CDDP whose T 50 =4.1 min (released 50%) and T 90 =8.0 min (released 90%).

[Comparison between the qualitative similarity and the quantitative similarity of chromatographic fingerprints of traditional Chinese medicines].

To explore the differences between the qualitative similarity and the quantitative similarity of chromatographic fingerprints of traditional Chinese medicines, the quantitative similarity calculated by vector shadow C%, apparent quantitative similarity R%, quantitative similarity P%, etc. were firstly proposed to disclose the quantitative information characteristics of traditional Chinese medicines fingerprints. The HPLC fingerprints of both the standard Fructus gardeniae and the ten batches of Fructus gardeniae produced in different places were evaluated by the new parameters to obtain good results. The contrasted fingerprint contained 35 peaks while geniposide was selected as the reference peak. The HPLC fingerprint had good precision and reproducibility with the RSD of the relative retention time less than 1.5% and the RSD of the relative peak area within 5%. The qualitative similarity and quantitative similarity between each crude drug and the contrasted fingerprint were quantitatively calculated, the values of C%, P%, etc., were applied in the quality control practice, which had less errors. What is more, this method could be used for the overall quality control of Fructus gardeniae and especially suits for qualitative and quantitative evaluations of the chromatographic fingerprints both in chemical constituent distribution and in contents. The quantitative parameters such as C% and P% can be used to objectively, authentically and thoroughly display the content information characteristics. When they combined with the qualitative similarity, it will be the good method to evaluate the chromatographic fingerprints of traditional Chinese medicines.

[The digital criterion of the potential information characteristics of the traditional Chinese medicine chromatographic fingerprints].

AIM: To exploit the characteristic digital criterion for the potential information characteristics of traditional Chinese medicine chromatographic fingerprints, the 37 parameters such as F and I were firstly proposed to disclose the potential information characteristics of traditional Chinese medicine fingerprints. METHODS: The HPLC fingerprints of the Ginkgo biloba extract (GBE) , Ginkgo leaf extract and diphyridamole injection (GLEDI), Ixeris sonchifolia Hance (ISH) and Ixeris sonchifolia Hance injection (ISHI) were compared each other. RESULTS: As far as the peak signal intensity, the uniform of peak signal, resolution and the fingerprint information were concerned. The GBE fingerprint was better than the GLEDI's, and the ISH fingerprint was also better than the ISHI's, then GBE fingerprint was close to the ISHI' s. CONCLUSION: The 37 parameters such as F and I can be used to objectively, authentically and thoroughly display the potential information characteristics of the traditional Chinese medicine chromatographic fingerprints.

[Study on the fingerprints of Fructus Forsythiae by high performance capillary electrophoresis].

The capillary electrophoresis fingerprints (CEFP) of Fructus Forsythiae was established to control its quality. The background electrolyte (BGE) was a 75 mmol/L sodium borate solution adjusted to pH 9.7 with 0.1 mmol/L NaOH solution. The detection wavelength was 228 nm and a voltage of 14 kV was applied. The Forsythia suspensa (Thunb.) Vahl was extracted by water and a set of CEFP containing 29 peaks was obtained. The CEFP had acceptable precision and reproducibility with the relative standard deviation (RSD) of the relative migration time less than 5.0% and the RSD of the relative peak area from 2% to 15%. There were good similarities between the standard CEFP and each set of CEFP of Forsythia suspensa (Thunb.) Vahl from ten different places, and their similarity coefficients were not less than 0.94. The CEFP were also evaluated by the information index I and the relative information index Ir, of chromatographic fingerprint.