Gut microbiota dysbiosis in patients with hepatitis B virus-related cirrhosis

Introduction and aimChronic hepatitis B (CHB) is a global epidemic disease that results from hepatitis B virus (HBV) infection and may progress to liver cirrhosis. The relationship between hepatitis B virus-related cirrhosis (HBV-RC) and gut microbiota dysbiosis is still unclear. The aim of this study is to elucidate the compositional and functional characteristics of the gut microbiota in the patients with liver cirrhosis and healthy individuals.Materials and methodsWe analyzed the gut microbiome in patients with HBV-RC and healthy individuals by 16S rRNA sequencing and metagenomic sequencing of fecal samples. A total of 113 genera, 85 families, 57 orders, 44 classes and 21 phyla were performed.ResultsOur results suggests that the composition of the gut microbiota had changed in the early stages of cirrhosis. We further identified more than 17 genera with different richness in compensated and decompensated cirrhosis groups. PICRUSt analysis showed that changes in bacterial composition can lead to significant changes in gene function, which may be one of the causes of liver cirrhosis.ConclusionOur study demonstrated that the composition of gut microbiota changed at different phases of HBV-RC. Gut microbiome transformation may be a biological factor in the progression of cirrhosis.     

Comparison of toxic reaction of Tripterygium wilfordii multiglycoside in normal and adjuvant arthritic rats

Aim of the study

Tripterygium wilfordii multiglycoside (GTW), an authorized Chinese patent drug, is used for treatment of rheumatoid arthritis and other immune disease. This study was to determine whether GTW induced different toxic reactions in adjuvant arthritis rats (AA rats) compared to those in normal rats.

Materials and methods

To prepare arthritic rat model, male Sprague-Dawley (SD) rats were immunized by injecting complete Freund's Adjuvant into right hind footpad. And then, GTW was given to rats intragastrically at dosage of 7 or 105 mg kg⁻¹ day⁻¹ from day 15 to day 28 after immunization. Routine clinical parameters and histopathologic changes of liver, kidney and testis were examined. Metabolic profiling in serum of groups was analyzed by LC-MS. A principal component analysis (PCA) and partial-least-squares discriminate analysis (PLS-DA) were carried out combined with mass spectrometry (MS) data set. All the quantitative data were performed by two-way ANOVA analysis following Student's t-test.

Results and conclusions

Treatment with GTW at both doses could diminish the right and left hind paws swelling. There was slight lipoid degeneration in hepatic tissue of normal rats treated by high dose of GTW, but there were not distinctly pathological changes in hepatic tissue of AA rats treated by GTW. Compared normal rats administered with GTW, no statistically significant difference in the serum alanine aminotransferase (ALT), creatinine (CRE), and blood urea nitrogen (BUN) levels were observed. However, the serum aspartate aminotransferase (AST) level was significant decreased in AA rats under exposure GTW compared with normal rats in the same conditions (p < 0.05), which indicated that GTW could offer a different liver toxic reaction in normal and AA rats. The metabolic analysis showed that a clear separation of PCA and PLS-DA score spot in normal rats, but not separation was seen in AA rats perturbed with low dosage GTW. The result indicated low dosage GTW might arouse a general toxic in normal rats but not in AA rats. The biomarker analysis showed that the level of lysophosphatidylcholines (LPCs) was down-regulated, but the level of ursodeoxycholic acid (UDCA) and chenodexycholic acid (CDCA) was up-regulated in AA rats compared with normal rats under exposure GTW. According to pathway analysis of metabolic markers, we conceived that LPC, UDCA and CDCA were the critical intermediates of choline and fatty acid metabolism. And the lipid metabolism was a correlative outcome of GTW induced toxicity in the liver in physiological condition animals. Taken together, GTW could induce different toxic reactions between normal and AA rats, and the lipid metabolism might be part of the mechanism for the hepatic lipidosis or the other liver injury.     

A metabolomic approach to lung cancer

Lung cancer is one of the most common cancers in the world, but no good clinical markers that can be used to diagnose the disease at an early stage and predict its prognosis have been found. Therefore, the discovery of novel clinical markers is required. In this study, metabolomic analysis of lung cancer patients was performed using gas chromatography mass spectrometry. Serum samples from 29 healthy volunteers and 33 lung cancer patients with adenocarcinoma (n = 12), squamous cell carcinoma (n = 11), or small cell carcinoma (n = 10) ranging from stage I to stage IV disease and lung tissue samples from 7 lung cancer patients including the tumor tissue and its surrounding normal tissue were used. A total of 58 metabolites (57 individual metabolites) were detected in serum, and 71 metabolites were detected in the lung tissue. The levels of 23 of the 58 serum metabolites were significantly changed in all lung cancer patients compared with healthy volunteers, and the levels of 48 of the 71 metabolites were significantly changed in the tumor tissue compared with the non-tumor tissue. Partial least squares discriminant analysis, which is a form of multiple classification analysis, was performed using the serum sample data, and metabolites that had characteristic alterations in each histological subtype and disease stage were determined. Our results demonstrate that changes in metabolite pattern are useful for assessing the clinical characteristics of lung cancer. Our results will hopefully lead to the establishment of novel diagnostic tools.     

牡丹皮炭给药后血清代谢物的~1H-NMR代谢组学研究

目的运用1H-NMR代谢组学方法研究正常人服用牡丹皮炭前后的血清代谢组变化及探讨牡丹皮炭的临床作用机制。方法采用磁共振技术对人体的血清进行测试分析,应用偏最小二乘判别分析法(PLS-DA)研究比较正常人服用牡丹皮炭前后血清之间的代谢谱差异。结果服用牡丹皮炭后,血清中变化最明显的为葡萄糖含量下降,乳酸和糖蛋白含量升高。结论正常人服用牡丹皮炭前后的血清在1H-NMR中出现的特征峰有明显差异,为医学临床检验和临床应用前期研究提供了一个参考。     

基于主成分聚类和PLS回归分析的穿心莲质量等级评价研究

目的结合性状指标和内在质量指标对穿心莲进行质量等级评价,建立等级评价模型,为穿心莲及其他中药材等级标准的制定提供参考。方法采收不同批次的穿心莲药材样品,测量和描述穿心莲药材的性状,采用数量分类学的方法对其评价指标进行编码,以HPLC法测定药材的4个二萜内酯类成分的含量,将编码后的性状指标和4个内酯的总量及醇溶性浸出物含量做相关性分析,初步筛选出13个评价指标,应用主成分聚类分析法对这13个指标进行分析,划分等级;并以偏最小二乘-判别分析(PLS-DA)对所划分的等级进行判别分析;最后,通过偏最小二乘回归分析(PLS)建立穿心莲药材等级的预测模型。结果根据主成分聚类分析法可以将穿心莲药材划分为3个等级,PLS-DA分析表明该等级划分合理;PLS回归分析所建立的穿心莲质量等级的预测模型为药材等级(Y)=3.761-0.020×叶含量-0.388×穿心莲内酯含量-1.117×新穿心莲内酯含量-0.274×去氧穿心莲内酯含量-0.287×脱水穿心莲内酯含量-0.302×4个内酯总量-0.104×醇溶性浸出物含量-0.015×茎颜色-0.0084×叶颜色-0.003×茎基部直径+0.020×分枝数+0.137×茎上部直径+0.011×株高,若Y在0.7～1.3,则预测穿心莲药材为一等品;若Y在1.7～2.3,则为二等品;若Y在2.7～3.3则为合格品。结论主成分聚类分析法结合PLS回归建立的穿心莲等级快速评价模型评价效果较为理想,可作为穿心莲药材质量等级的快速评价模型,为穿心莲及其他中药材质量等级评价及其等级标准的制定提供新思路。     

基于UPLC-Triple TOF-MS/MS技术分析何首乌代谢物积累的动态变化

目的分析不同采收时间何首乌Polygonum multiflorum代谢产物积累的动态变化。方法采用超高效液相色谱-串联四极杆飞行时间高分辨质谱(UPLC-Triple TOF-MS/MS)技术对不同采收时间何首乌15批样品进行测定,通过多级串联质谱分析,对其质谱数据进行峰匹配、峰对齐、滤噪处理等进行特征峰提取;采用主成分分析(PCA)和偏最小二乘法-判别分析(PLS-DA)进行数据处理;通过一级质谱精确质荷比和二级质谱碎片信息,结合软件数据库搜索及相关文献进行成分鉴别。结果不同采收时间何首乌样品间的化学组成得到明显区分;初步筛选出不同采收时间何首乌样品间25种差异显著的化学成分并鉴定出24个成分,其中共有9种差异化学成分呈现不同的变化规律。结论为揭示不同采收时间何首乌中代谢物积累动态规律及确定其药材合理采收期提供基础资料。     

Toxicological effects of cinnabar in rats by NMR-based metabolic profiling of urine and serum

Cinnabar, an important traditional Chinese mineral medicine, has been widely used as a Chinese patent medicine ingredient for sedative therapy. However, the pharmaceutical and toxicological effects of cinnabar, especially in the whole organism, were subjected to few investigations. In this study, an NMR-based metabolomics approach has been applied to investigate the toxicological effects of cinnabar after intragastrical administration (dosed at 0.5, 2 and 5 g/kg body weight) on male Wistar rats. Liver and kidney histopathology examinations and serum clinical chemistry analyses were also performed. The 1H NMR spectra were analyzed using multivariate pattern recognition techniques to show the time- and dose-dependent biochemical variations induced by cinnabar. The metabolic signature of urinalysis from cinnabar-treated animals exhibited an increase in the levels of creatinine, acetate, acetoacetate, taurine, hippurate and phenylacetylglycine, together with a decrease in the levels of trimethyl-N-oxide, dimethylglycine and Kreb's cycle intermediates (citrate, 2-oxoglutarate and succinate). The metabolomics analyses of serum showed elevated concentrations of ketone bodies (3-d-hydroxybutyrate and acetoacetate), branched-chain amino acids (valine, leucine and isoleucine), choline and creatine as well as decreased glucose, lipids and lipoproteins from cinnabar-treated animals. These findings indicated cinnabar induced disturbance in energy metabolism, amino acid metabolism and gut microflora environment as well as slight injury in liver and kidney, which might indirectly result from cinnabar induced oxidative stress. This work illustrated the high reliability of NMR-based metabolomic approach on the study of the biochemical effects induced by traditional Chinese medicine.     

基于抗菌活性及病理指标评价子芩与枯芩对肺炎的药效学差异

目的比较子芩与枯芩(均为黄芩的根)体外抗菌活性及体内对肺炎大鼠的药效学差异。方法采用体外实验,以二倍微量稀释法测定子芩与枯芩对肺炎链球菌的最小抑菌浓度(MIC);体内实验以肺炎大鼠为模型,选择体温、肺脏指数、血常规检测以及大鼠肺部病理切片为评价指标评价子芩与枯芩对肺炎大鼠作用的药效学差异;采用偏最小二乘-判别分析(PLS-DA)法,对子芩和枯芩的抗菌活性及病理指标进行分析。结果子芩对肺炎链球菌的MIC为0.5 g/mL,枯芩的MIC为0.25 g/mL;子芩和枯芩均能显著降低肺炎大鼠的体温、肺脏指数、全血中白细胞数、中性粒细胞数和淋巴细胞数;PLS-DA分析显示子芩与枯芩的体外抗菌活性及体内抗肺炎活性具有显著差异。结论子芩与枯芩对肺炎均具有一定的治疗作用,两者的作用强度有明显差异,枯芩作用优于子芩。     

Determination of Residence Time Distribution in a Continuous Powder Mixing Process With Supervised and Unsupervised Modeling of In-line Near Infrared (NIR) Spectroscopic Data

Successful implementation of continuous manufacturing processes requires robust methods to assess and control product quality in a real-time mode. In this study, the residence time distribution of a continuous powder mixing process was investigated via pulse tracer experiments using near infrared spectroscopy for tracer detection in an in-line mode. The residence time distribution was modeled by applying the continuous stirred tank reactor in series model for achieving the tracer (paracetamol) concentration profiles. Partial least squares discriminant analysis and principal component analysis of the near infrared spectroscopy data were applied to investigate both supervised and unsupervised chemometric modeling approaches. Additionally, the mean residence time for three powder systems was measured with different process settings. It was found that a significant change in the mean residence time occurred when comparing powder systems with different flowability and mixing process settings. This study also confirmed that the partial least squares discriminant analysis applied as a supervised chemometric model enabled an efficient and fast estimate of the mean residence time based on pulse tracer experiments.     

基于电子舌的白及及其近似饮片的快速辨识研究＊

目的 探讨电子舌方法用于白及及其近似饮片快速辨识的可行性。方法 收集45批白及饮片及其近似品天麻饮片30批、玉竹饮片30批、黄花白及饮片29批，分别进行药典与地方标准辨识（M₁法）、HPLC指纹图谱辨识（M₂法），并结合原始采购信息获取最终饮片种类的标杆信息（Y），再采集电子舌味觉感官数据（X）并利用化学计量学方法分别建立主成分分析-判别分析（PCA-DA）、偏最小二乘-判别分析（PLS-DA）的45批白及饮片与剩余89批饮片的二分类辨识模型和45批白及饮片、30批天麻饮片、30批玉竹饮片、29批黄花白及饮片的四分类辨识模型（Y=F（X），M₃法）。结果 经留一法交互验证，基于PCA-DA、PLS-DA二分类辨识模型的正判率分别为98.51%、100.00%，基于PCA-DA、PLS-DA四分类辨识模型的正判率分别为100.00%（无未分类样本）、100.00%（有4个未分类样本），模型判别良好，结合正判率与模型未分类样本数两项指标，最终选择二分类辨识以PLS-DA为最终辨识模型、四分类辨识以PCA-DA为最终辨识模型，两种模型正判率均为最高，且均未出现未分类样本。结论 电子舌可快速准确辨识白及及其近似饮片，为未来研发智能化中药饮片快速辨识设备提供了思路。