Metabolomic Markers in Tongue-Coating Samples from Damp Phlegm Pattern Patients of Coronary Heart Disease and Chronic Renal Failure.

In this paper, we used tongue coating to obtain metabolites in patients with coronary heart disease (CHD) and chronic renal failure (CRF). The metabolites were analyzed to discover the substance that serves as the underlying basis of the damp phlegm pattern. This analysis is based on the Traditional Chinese Medicine (TCM) theory of "different diseases have the same pattern." The metabolic spectrum was obtained through the Gas Chromatography Mass Spectrometry coupling techniques and analyzed by searching the METLIN and HMDB databases. Some metabolites related to amino acids and glucose metabolism were identified in the tongue-coating samples from damp phlegm pattern patients by comparing them to nondamp phlegm pattern patients and healthy subjects. In addition, there were five common metabolites in the tongue-coating samples from CHD damp phlegm pattern patients compared to CRF damp phlegm pattern patients, which allowed us to understand the theory of "different diseases have the same pattern." In the future, the metabolites identified in this study may be used as noninvasive and convenient biomarkers to distinguish the damp phlegm pattern of CHD and CRF patients.

Non-targeted metabolomics reveals diagnostic biomarker in the tongue coating of patients with chronic gastritis.

Analysis of the properties of the tongue has been used in traditional Chinese medicine for disease diagnosis. Notably, tongue analysis, which is non-invasive and convenient compared with gastroscopy and pathological examination, can be used to assess chronic gastritis (CG). In order to find potential diagnostic biomarkers and study the metabolic mechanisms of the endogenous small molecules in the tongue coating related to CG, a non-targeted metabolomic analysis method was developed using ultra high performance liquid chromatography combined with quadrupole time-of-flight mass spectrometry (UHPLC-Q/TOF-MS). It was performed using two different columns in positive and negative ion scanning modes separately. The stability of the samples was evaluated and the age and gender factors of the subjects were excluded to ensure the reliability of the data in this study. Finally, under the four analysis models, 130, 229, 113 and 92 differential compounds were found using multivariate statistical methods respectively. 37 potential biomarkers were putatively identified after removing the duplicate compounds and five potential diagnostic biomarkers were putatively identified by receiver operating characteristic (ROC) curve analysis, including inosine, oleamide, adenosine, N-acetylglucosamine (GlcNAc) and xanthine. The main metabolic pathways associated with CG were purine metabolism, amino acid metabolism, sphingolipid metabolism and energy metabolism, which suggested that oxygen free radicals and energy metabolism were altered in patients with CG. These results provided a potential new basis for the quantitative diagnosis and pathogenesis of CG.

A historical evaluation of Chinese tongue diagnosis in the treatment of septicemic plague in the pre-antibiotic era, and as a new direction for revolutionary clinical research applications.

Chinese tongue diagnosis was initially developed to quickly and efficiently diagnose and prescribe medicine, while at the same time allowing the doctor to have minimal contact with the patient. At the time of its compiling, the spread of Yersinia pestis, often causing septicaemia and gangrene of the extremities, may have discouraged doctors to come in direct contact with their patients and take the pulse. However, in recent decades, modern developments in the field of traditional Chinese medicine, as well as the spread of antibiotics in conjunction with the advancements of microbiology, have overshadowed the original purpose of this methodology. Nevertheless, the fast approaching post-antibiotic era and the development of artificial intelligence may hold new applications for tongue diagnosis. This article focuses on the historical development of what is the world's earliest tongue diagnosis monograph, and discusses the directions that such knowledge may be used in future clinical research.

Biology of the Tongue Coating and Its Value in Disease Diagnosis.

Tongue diagnosis is one of the most important diagnostic tools in traditional Chinese medicine and has been verified for thousands of years. However, its subjectivity and repeatability has been disputed continuously. The tongue coating as the primary coverage of tongue diagnosis provides more objectivity and reproducibility due to its relatively clear molecular basis; it also has a close relationship with many system diseases and may be used as a potentially valuable disease diagnostic tool. This article describes the material basis of the tongue coating, including its biology (epithelial cells, blood cells, vascular endothelial cells, and bacteria) and its metabolites; moreover, we summarize the diseases that are most correlated with the tongue coating. This will be valuable not only for fundamental research of tongue diagnosis but also for the diagnosis and differential diagnosis of disease. We suppose that the tongue coating could serve as a valuable auxiliary diagnosis tool in many diseases, and more research should focus on how to colligate the various information about the tongue and provide useful information for disease diagnosis.

[Purple-bluish tongue is associated with platelet counts, and the recurrence of epithelial ovarian cancer].

OBJECTIVE: To evaluate the relationship between purple-bluish tongue and platelet counts, and further to examine their associations with the recurrence of epithelial ovarian cancer. METHODS: A total of 82 epithelial ovarian cancer patients were enrolled in this study. Cluster analysis was used for grouping patients' P(RGB) (Red-R; Green-G; Blue-B; Average percentage of RGB, P(RGB)) values. Receiver operating characteristic (ROC) curve was performed for detecting the diagnostic standard of purple-bluish tongue. Χ2 test was used to assess the relationship between purple-bluish tongue and platelet counts, and the recurrence of epithelial ovarian cancer. The perioperative (preoperative) platelet level was examined with tongue image and disease recurrence. RESULTS: Tongue images were classified into two groups basing on P(RGB) values of images by cluster analysis. The numbers of cases in cluster "1" (normal color tongue) was 16 and cluster "2" (purple-bluish tongue) was 66. Two groups of P(RGB) values, classified by cluster analysis, were significantly correlated with vision-based tongue color recognition (Kappa = 0.852, P < 0.001). ROC curve showed that the ratio of P(B) to P(R) had the highest diagnostic value. The sensitivity and the specificity of the ratio of P(B) to P(R) were 95.3% and 88.9% respectively and the optimal cut-off point was 0.71. Purple-bluish tongue was significantly correlated with increased platelet counts (P < 0.001). Both the increased platelet counts (P = 0.01) and purple-bluish tongue were associated with recurrence of epithelial ovarian cancer (P < 0.001). CONCLUSION: The ratio of P(B) to P(R) greater than 0.71 could serve as an indicator for purple-bluish tongue diagnosing used in symptom pattern identification in Traditional Chinese Medicine. Purple-bluish tongue, associated with increased platelet counts, was also closely correlated with the recurrence of epithelial ovarian cancer.

Reliability and validity of tongue color analysis in the prediction of symptom patterns in terms of East Asian Medicine.

OBJECTIVE: To examine whether color parameters of tongue inspection (TI) using a digital camera was reliable and valid, and to examine which color parameters serve as predictors of symptom patterns in terms of East Asian medicine (EAM). METHODS: Two hundred female subjects' tongue substances were photographed by a mega-pixel digital camera. Together with the photographs, the subjects were asked to complete Yin deficiency, Phlegm pattern, and Cold-Heat pattern questionnaires. Using three sets of digital imaging software, each digital image was exposure- and white balance-corrected, and finally L* (luminance), a* (red-green balance), and b* (yellow-blue balance) values of the tongues were calculated. To examine intra- and inter-rater reliabilities and criterion validity of the color analysis method, three raters were asked to calculate color parameters for 20 digital image samples. Finally, four hierarchical regression models were formed. RESULTS: Color parameters showed good or excellent reliability (0.627-0.887 for intra-class correlation coefficients) and significant criterion validity (0.523-0.718 for Spearman's correlation). In the hierarchical regression models, age was a significant predictor of Yin deficiency (ß = 0.192), and b* value of the tip of the tongue was a determinant predictor of Yin deficiency, Phlegm, and Heat patterns (ß = - 0.212, - 0.172, and - 0.163). Luminance (L*) was predictive of Yin deficiency (ß = -0.172) and Cold (ß = 0.173) pattern. CONCLUSION: Our results suggest that color analysis of the tongue using the L*a*b* system is reliable and valid, and that color parameters partially serve as symptom pattern predictors in EAM practice.

Immediate effects of acupuncture on tongue pressure including swallowing reflex latency in Parkinson's disease.

BACKGROUND: It is important to evaluate tongue function in terms of its clinical implications for swallowing ability. Motor dysfunction and loss of coordination of the tongue are frequently seen, and this influences the oral and pharyngeal phases of swallowing. The purpose of this pilot study was to evaluate the effect of a single acupuncture treatment for tongue pressure in Parkinson's disease. METHODS: A total of 13 patients, aged 57- 84 years, were recruited. Tongue pressure was measured using a tongue pressure manometer. Furthermore, swallowing reflex latency was measured in 3 of the 13 patients. RESULTS: Significant changes were seen after acupuncture in mean tongue pressure, which increased from 23.1 to 26.7 kPa (p<0.01). Reductions were seen after acupuncture in mean swallowing reflex latency (from 5.2 to 4.6 s for first saliva swallow; from 19.9 to 15.7 s for second saliva swallow; and from 10.4 to 5.7 s for third saliva swallow(s)). CONCLUSIONS: Our study's findings suggest that acupuncture may be useful for improving oral cavity function, but further controlled trials are needed.

The association between objective tongue color and endoscopic findings: results from the Kyushu and Okinawa population study (KOPS).

BACKGROUND: The relation between tongue color and gastroesophageal disease is unclear. This study was done to investigate the associations between tongue color (TC), endoscopic findings, Helicobacter.pylori infection status, and serological atrophic gastritis (SAG). METHODS: The participants were 896 residents of Ishigaki Island, Okinawa, aged 28-86 years. The tongue was photographed, esophagogastroduodenoscopy was done, and serum antibody to H.pylori was measured. SAG was defined as a serum Pepsinogen (PG)Ilevel ≤70 ng/ml and a PGI/IIratio ≤3.0. TC was measured by the device-independent international commission on Illumination 1976 L*a*b* color space standards at four points: (1) edge, (2) posterior, (3) middle, and (4) apex. We also calculated the ratio of the tongue edge to the three other measured points to examine the association between the coating of the tongue and the endoscopic and laboratory findings. RESULTS: Participants were excluded who had two or more endoscopic findings (n = 315) or who had SAG without seropositivity to H.pylori (n = 33). The remaining 548 participants were divided into three groups: SAG and seropositive to H.pylori (n = 67), seropositive to H.pylori alone (n = 56), and without SAG and seronegative for H.pylori (n = 425). We divided 425 residents into a single endoscopic finding positive group (n = 207) and a negative group, which served as a control (n = 218). The most frequent single endoscopic finding was esophageal hernia (n = 110), followed by erosive esophagitis (n = 35) and erosive gastritis (EG) (n = 45). EH was significantly associated with TC (2b*/1b*) (P < 0.05). EG was significantly associated with TC (3a*, 3b*) (P < 0.05). Seropositivity to H.pylori was significantly associated with TC (3 L*, 3 L*/1 L*) (P < 0.05, <0.01), and seropositivity to both H.pylori and SAG was significantly associated with TC (3 L*/1 L*) (P < 0.05). Multivariate analysis extracted TC (3a*, 3b*) as an independent factor associated with a differential diagnosis of EG (Odds ratio (OR) 2.66 P = 0.008, OR 2.17 P = 0.045). CONCLUSIONS: The tongue body color of the middle area reflects acute change of gastric mucosa, such as erosive gastritis. Tongue diagnosis would be a useful, non-invasive screening tool for EG.

Properties of oil/water emulsions affecting the deposition, clearance, and after-feel sensory perception of oral coatings.

The aims of this study were to investigate the influence of (i) protein type, (ii) protein content, and (iii) viscosity of o/w emulsions on the deposition and clearance of oral oil coatings and after-feel perception. Oil fraction (m(oil)/cm(2)(tongue)) and after-feel perception differed considerably between emulsions which do not flocculate under in mouth conditions (Na-caseinate) and emulsions which flocculate under in mouth conditions (lysozyme). The irreversible flocculation of lysozyme stabilized emulsions caused slower oil clearance from the tongue surface compared to emulsions stabilized with Na-caseinate. Protein content had a negative relation with oil fraction for lysozyme stabilized emulsions and no relation for Na-caseinate stabilized emulsions immediately after expectoration. Viscosity differences did not affect oil fraction, although the presence of thickener decreased deposition of oil on tongue. We conclude that after-feel perception of o/w emulsions is complex and depends on the deposited oil fraction, the behavior of proteins in mouth, and thickeners.

Physical and sensory characterizations of oral coatings of oil/water emulsions.

The physical and sensory properties of oil coatings on the tongue formed by five oil/water emulsions varying in oil content were investigated. A total of 20 subjects processed orally each emulsion for 30 s in triplicate. In vivo fluorescence measurements at the front and back of the anterior tongue were made to quantify the oil fraction deposited at different time points. Calibration lines relating fluorescence intensity to oil fraction were determined using pig tongues at 37.5 °C to mimic oral conditions. The oil fraction on the tongue increased linearly with an increasing oil content of the emulsions. The oil fraction deposited at the back of the anterior tongue was 1.5-2.0× larger than at the front. The intensity of sensory attributes describing after-feel perception was related to the oil fraction by Weber-Fechner's law. This study uses in vivo fluorescence to study food behavior in the mouth and unravel new insights in after-feel perception of emulsions.

Metabolic markers and microecological characteristics of tongue coating in patients with chronic gastritis.

BACKGROUND: In Traditional Chinese Medicine (TCM), tongue diagnosis has been an important diagnostic method for the last 3000 years. Tongue diagnosis is a non-invasive, simple and valuable diagnostic tool. TCM treats the tongue coating on a very sensitive scale that reflects physiological and pathological changes in the organs, especially the spleen and stomach. Tongue coating can diagnose disease severity and determine the TCM syndrome ("Zheng" in Chinese). The biological bases of different tongue coating appearances are still poorly understood and lack systematic investigation at the molecular level. METHODS: Tongue coating samples were collected from 70 chronic gastritis patients and 20 normal controls. 16S rRNA denatured gradient gel electrophoresis (16S rRNA-DGGE) and liquid chromatography and mass spectrometry (LC-MS) were designed to profile tongue coatings. The statistical techniques used were principal component analysis and partial least squares-discriminate analysis. RESULTS: Ten potential metabolites or markers were found in chronic gastritis patients, including UDP-D-galactose, 3-ketolactose, and vitamin D2, based on LC-MS. Eight significantly different strips were observed in samples from chronic gastritis patients based on 16S rRNA-DGGE. Two strips, Strips 8 and 10, were selected for gene sequencing. Strip 10 sequencing showed a 100% similarity to Rothia mucilaginosa. Strip 8 sequencing showed a 96.2% similarity to Moraxella catarrhalis. CONCLUSIONS: Changes in glucose metabolism could possibly form the basis of tongue coating conformation in chronic gastritis patients. The study revealed important connections between metabolic components, microecological components and tongue coating in chronic gastritis patients. Compared with other diagnostic regimens, such as blood tests or tissue biopsies, tongue coating is more amenable to, and more convenient for, both patients and doctors.

[Metabolite changes in the greasy tongue coating of patients with chronic gastritis].

OBJECTIVE: To explore the changes in metabolites in the greasy tongue coating in patients with chronic gastritis. METHODS: Forty chronic gastritis patients presenting with greasy tongue coating, 30 chronic gastritis patients presenting with non-greasy tongue coating, and 20 healthy control persons presenting with light red tongues and thin white coating were enrolled, and the tongue coating was detected by combining artificial diagnosis and the Z-BOX Tongue Digital Analyzer's diagnosis. Samples of all the tongue coatings were collected before treatment. The metabolic fingerprinting of the tongue coating samples was obtained using ultra-performance liquid chromatography and mass spectrometry (UPLC-MS), and the metabolic components in the tongue coating samples were detected. After this, principal component analysis, partial least squares discriminant analysis and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to identify potential metabolic markers. Finally, the components were identified using the Chemspider and HMDB searching. RESULTS: UPLC-MS results were analyzed by OPLS-DA and showed that the metabolites among the three groups were distributed in different regions. The different potential metabolic markers between the patients with or without greasy coating were 3-ketolactose, 2-deoxy-D-ribose, UDP-D-galactose metarhodopsin, ascorbate, picolinate and histidine. The different potential metabolic markers between the greasy coating group and the normal group were 3-ketolactose, UDP-D-galactose, leukotriene A4 and vitamin D(2). CONCLUSION: The metabolites of the greasy coating group, the non-greasy coating group and the normal group show significant differences in energy metabolism, mainly of glucose metabolism. This demonstrated that glucose metabolism may be one of the mechanisms leading to the formation of greasy coating.

Tablets containing a cysteine protease, actinidine, reduce oral malodor: a crossover study.

Tongue coating (TC) mainly consists of protein mostly from exfoliated epithelial cells. Until now, to reduce TC accumulation, only mechanical measures have been available, and the procedure involves unpleasant side effects, such as gagging reflex or carcinogenesis related to mechanical stimulation. We expected that protease might be effective in reducing the accumulation of TC causing oral malodor. The purpose of this study was to determine the effect of long-term use of candy tablets containing protease, actinidine, on both TC accumulation and concentration of volatile sulfur compounds (VSCs) in mouth air. We employed 14 subjects aged 24 to 54 years old for this study, and conducted a double-blind randomized crossover trial. The subjects sucked the tablets containing actinidine three times a day until the sixth day after starting the study. The tablets without actinidine were utilized as a placebo. Measurements of VSC concentration and TC accumulation were carried out before and after chewing tablets on the first day, and also on the seventh day. The levels of VSC and TC significantly (p < 0.05) decreased after tablets were taken on the first day in both the test and placebo groups. There was a statistically significant decrease (p < 0.05) in VSC after seven days of use only in the test group. The results of the study suggest that the tablets containing actinidine had an accumulative effect in reducing VSC in mouth air with long-term use.

The effect of various forms of selenium supplied to pregnant goats on the levels of selenium in the body of their kids at the time of weaning.

The aim of this trial was to compare the effect of long-term supplementation of goats with different forms of selenium on body reserves of selenium in their kids at the time of weaning. Thirty-three pregnant goats were divided into five groups. Group C was control while the other four groups were supplemented with selenium (Se) for 6 weeks before parturition (0.3 mg/goat/day) and after parturition (0.9 mg/goat/day). Group "Se-I" received sodium selenite and three other groups received organic forms: "Se-L," lactate-protein complex; "Se-P," Se-proteinate; and "Se-Y," Se-yeast. The kids were weaned at 3 months of age and samples of tissues (liver, pancreas, myocardium, lungs, kidneys, spleen, thigh, tongue, and diaphragm) were taken after slaughtering. The long-term supplementation of goats with Se influenced Se concentration in all examined tissues of kids. Significant differences (p≤0.01) were found between the control and all experimental groups, except for the renal cortex and pancreas (Se-I). The average increase of Se concentration in overall examined tissues in comparison with the control (100%) was as follows: Se-Y, 192%; Se-P, 167%; Se-L, 161%; Se-I, 144%. The highest efficiency was found in the group supplemented with Se-yeast with a high content of selenomethionine, also the other two organic forms of Se were more efficient than the inorganic form.

Study on relationship between the thickness of tongue fur and the expressions of apoptosis-related genes of the tongue epithelial cells in patients with diseases of the digestive system.

To investigate the relationship between the thickness of tongue fur, apoptosis of the tongue fur epithelial cells and expressions of apoptosis-related genes in diseases of the digestive system, apoptosis-related genes TGF-beta3, fas mRNA and protein products were detected with terminal deoxynucleotidyl transferase-mediated deoxyurine triphosphate (d-UTP) nick-end labeling (TUNEL) technique, in situ hybridization, immunohistochemical methods, and image analysis technique, respectively. Results indicated that compared with the normal tongue fur, over-expression of fas gene was found in the peeling fur with an increase in cell apoptosis, while a low-expression of TGF-beta3 in the thick fur with a decrease in cell apoptosis. The changes in expression levels of fas and TGF-beta3 genes, apoptosis-promoting genes in the tongue fur epithelial cells, had a similar tendency of cell apoptosis level. It is concluded that the changes in expression levels of fas and TGF-beta3 are possibly important reasons influencing apoptosis of epithelial cells of tongue fur and leading to changes in thickness of the tongue fur.

Fast-disintegrating sublingual tablets: effect of epinephrine load on tablet characteristics.

The aim of this study was to evaluate the effect of increasing epinephrine load on the characteristics of fast-disintegrating sublingual tablets for the potential emergency treatment of anaphylaxis. Four tablet formulations, A, B, C, and D, containing 0%, 6%, 12%, and 24% of epinephrine bitartrate, respectively, and microcrystalline cellulose:low-substituted hydroxypropyl cellulose (9:1), were prepared by direct compression, at a range of compression forces. Tablet weight variation, content uniformity, hardness, disintegration time, wetting time, and friability were measured for each formulation at each compression force. All 4 tablet formulations at each compression force were within the United States Pharmacopeia (USP) limits for weight variation and content uniformity. A linear increase in compression force resulted in an exponential increase in hardness for all formulations, a linear increase in disintegration and wetting times of A, and an exponential increase in disintegration and wetting times of B, C, and D. At a mean +/- SD hardness of > or = 2.3 +/- 0.2 kg, all tablet formulations passed the USP friability test. At a mean +/- SD hardness of < or = 3.1 +/- 0.2 kg, all tablet formulations resulted in disintegration and wetting times of <10 seconds and <30 seconds, respectively. Tablets with drug loads from 0% to 24% epinephrine can be formulated with hardness, disintegration times, and wetting times suitable for sublingual administration.

Selenium influences tissue levels of selenoprotein W in sheep.

Because selenium increases the levels of other selenoproteins, the influence of this element on selenoprotein W was examined in wether sheep fed either a low selenium diet (0.02 mg/kg) or the same diet supplemented with 3 mg selenium as selenite per kilogram diet. Muscle biopsies were taken initially and at 3.5, 7.0 and 10.5 wk. The sheep were killed after the last muscle biopsy and samples from nine tissues were taken. Selenoprotein W was determined in tissues by Western blots with a polyclonal antibody against a synthetic peptide based on the protein sequence of the homologous rat selenoprotein W. In supplemented sheep, muscle selenoprotein W was significantly increased over initial levels (P < 0.05) at 7 wk and afterwards, whereas in sheep consuming the low selenium diet, muscle selenoprotein W levels declined significantly (P < 0.05) after 10.5 wk. This selenoprotein was found in various amounts in all tissues examined. The highest levels of selenoprotein W were found in skeletal muscles and heart and the lowest was found in liver. Except for selenoprotein W in brain, the concentrations of selenoprotein W, selenium and glutathione peroxidase activity were significantly higher (P < 0.05) in all tissues from supplemented sheep than in those from unsupplemented sheep. The selenoprotein W levels in brains of the two groups were not significantly different. Thus, selenoprotein W levels in all tissues of sheep except the brain are sensitive to selenium status.

[Preliminary study of cytochemistry on thin-white coating of tongue with deficiency-cold syndrome].

The method of qualitative and quantitative analysis and localization of cytochemistry were used to observe the epithelial cells of the thin-white coating of tongue from 25 patients with Deficiency Cold Syndrome (DCS) and 20 normal persons. Results showed that the level of intracellular DNA, PAS of the epithelial cells in the DCS group were markedly enhanced, while the activities of succinate dehydrogenase (SDH) and acid alpha-naphthyl acetate esterase (ANAE) were reduced significantly compared to normal group, P < 0.05, 0.001, 0.001, 0.01 respectively.