Exploring Central Regulatory Effect of Chaishao Liujuntang on Chronic Atrophic Gastritis Rats with Liver Depression and Spleen Deficiency Syndrome Based on Metabolomics / 中国实验方剂学杂志

ObjectiveBased on ultra performance liquid chromatography-mass spectrometry（UPLC-MS） and non-targeted metabolomics technology to discuss the central regulatory effect of Chaishao Liujuntang on chronic atrophic gastritis（CAG） rats with liver-depression and spleen-deficiency， and to look for the correlation between cerebral cortex， hypothalamus and metabolic status of gastric tissues. MethodA CAG rat model with liver-depression and spleen-deficiency was established by chemical induction， hunger and satiety disorders， chronic restraint and tail clamping stimulation， lasting for 16 weeks. Twenty-eight Wistar rats were randomly divided into a blank group of 8 rats and a model group of 20 rats. After the completion of modeling， 4 rats in the model group were taken to observe the pathological changes of gastric mucosa. The remaining model rats were randomly divided into a model group of 8 rats and a Chaishao Liujuntang group of 8 rats. Chaishao Liujuntang group rats were given 5.1 g·kg-1 by gavage， and the remaining rats were given equal volume sterilized water by gavage for 4 weeks. Macroscopic characteristics， behavioral indicators and histopathological changes of the gastric mucosa of rats in each group were observed and compared. UPLC-MS non-targeted metabolomics was used to explore the metabolic regulation effect of Chaishao Liujuntang on the cerebral cortex， hypothalamus and stomach tissues of CAG rats with liver-depression and spleen-deficiency. Pearson correlation coefficient method was used to analyze the correlation between different tissue metabolites. ResultCompared with the model group， the macroscopic characteristics of rats in Chaishao Liujuntang group were improved， such as hair color， mental state and stool properties， and the number of times of crossing and standing in the open field experiment was significantly increased， and the static time of forced swimming was significantly reduced（P<0.01）， and the gastric mucosa atrophy was reduced. The metabolic data from the cerebral cortex of rats in each group identified a total of 3 common potential biomarkers， but not enriched in pathways， 26 common potential biomarkers were identified in the hypothalamus， and the key metabolic pathways involved were mainly enriched in purine metabolism， glycerol phospholipid metabolism， D-glutamine and D-glutamic acid metabolism. Seventeen common potential biomarkers were identified in the stomach， and the key metabolic pathways involved were mainly enriched in thiamine metabolism， valine， leucine and isoleucine biosynthesis， and taurine and taurine metabolism. Correlation analysis of metabolites in different tissues revealed that multiple amino acids and their derivatives mediated metabolic connections between the cerebral cortex， hypothalamus and stomach of rats. ConclusionThe metabolic disorders in the cerebral cortex， hypothalamus and stomach of CAG rats with liver-depression and spleen-deficiency have their own characteristics， mainly manifested by changes in the content of glycerol phospholipids， fatty acids and bile acid metabolites. Moreover， Chaishao Liujuntang may play a central regulatory role in CAG rats with liver-depression and spleen-deficiency by correcting the metabolic disorders of amino acids.

Altered gut microbiota-host bile acid metabolism in IBS-D patients with liver depression and spleen deficiency pattern.

BACKGROUND: Dysregulation of gut microbiota-host bile acid (BA) co-metabolism is a critical pathogenic factor of diarrhea-predominant irritable bowel syndrome (IBS-D). Traditional Chinese Medicine (TCM), instructed by pattern differentiation, is effective in treating IBS-D, in which liver depression and spleen deficiency (LDSD) is the most prevalent pattern. Still, it is unclear the linkage between the LDSD pattern and the BA metabolic phenotype. PURPOSE: This study aimed to uncover the biological basis of the LDSD pattern from the BA metabolic perspective. METHODS: Patients with IBS-D completed questionnaires regarding the irritable bowel severity scoring system (IBS-SSS), stool frequency, Stool Bristol scale, and Self-Rating Scales of mental health. Fasting blood and morning feces were collected to analyze the gut metagenome and BA-related indices/metabolites. RESULTS: IBS-D patients with LDSD had a higher incidence of BA overexcretion (41% vs. 23% non-LDSD) with significant elevations in fecal total BAs and serum BA precursor 7α-hydroxy-4-cholesten-3-one levels. Compared to controls or non-LDSD patients, LDSD patients had a featured fecal BA profile, with higher proportions of deoxycholic acid (DCA), 7-ketodeoxycholic acid, and lithocholic acid. It is consistent with the BA-metabolizing genomic changes in the LDSD gut microbiota characterized by overabundances of 7-dehydroxylating bacteria and BA-inducible genes (baiCD/E/H). The score of bowel symptoms (stool frequency and abdominal pain) showing greater severity in the LDSD pattern were positively correlated with bai-expressing bacterial abundances and fecal DCA levels separately. CONCLUSION: We clarified a differed BA metabolic phenotype in IBS patients with LDSD, which closely correlates with the severity of bowel symptoms. It demonstrates that gut microbiota and host co-metabolism of BAs would provide crucial insight into the biology of the LDSD pattern and its internal relationship with IBS progression.

HMGB1/PTEN/PI3K axis participates in the peripheral immune cell differentiation in two representative TCM syndromes of chronic hepatitis B patients.

Liver depression and spleen deficiency syndrome (LDSDS) and spleen-gastric damp-heat syndrome (SGDHS) are two major traditional Chinese medicine syndromes observed in chronic hepatitis B (CHB). Both syndromes exhibit significant differences in the pathogenesis and prognosis, and are closely related to the immune system. However, the underlying mechanisms are largely unknown. This study aimed to explore the immunoregulatory mechanisms of the two syndromes and promote the differentiation precision between the two syndromes. Thirty-six patients with CHB (18 LDSDS patients and 18 SGDHS patients) and 14 healthy controls were recruited into this study and blood was collected from all the subjects for testing. We studied the contents of T lymphocytes by flow cytometry and the expression levels of HMGB1/PTEN/PI3K axis proteins by enzyme-linked immunosorbent assay (Elisa). Protein-protein interaction (PPI) networks among HMGB1/PTEN/PI3K axis were constructed for functional enrichment. The correlations between T lymphocytes and proteins were analyzed by constructing multiple regression equations. The results revealed that the CD8+ T cells level in the two syndromes were lower than that in healthy controls, and the levels of Th17, Treg cells, and HMGB1, PI3K, PDK1, Akt were higher than those of the healthy controls (p < 0.05). Moreover, the levels of CD4+ T, Th17 cells, and HMGB1, PTEN, PI3K in LDSDS were higher than SGDHS (p < 0.05). PPI network indicated that HMGB1/PTEN/PI3K axis participated in T cell activation and liver pathology. Our results revealed that HMGB1/PTEN/PI3K axis may play an important role in regulating the formation of peripheral immune differences between the two syndromes. CD4+ T and Th17 are two representative immune cells that may serve as potential biological markers for LDSDS and SGDHS in CHB.

Clinical Effect of Chaishao Longmu Decoction on Insomnia in Patients with Liver Depression and Spleen Deficiency Based on Infrared Thermal Images / 中国实验方剂学杂志

ObjectiveTo observe the therapeutic effect of Chaishao Longmu decoction on insomnia in the patients with the syndrome of live depression and spleen deficiency and explore the correlation between infrared thermal imaging and insomnia with liver depression and spleen deficiency. MethodA total of 72 insomnia patients treated in the outpatient department of Shenzhen Hospital of Guangzhou University of Chinese Medicine （Futian） from May to December in 2022 were selected and randomized into a treatment group and a control group， with 36 patients in each group. The patients in the treatment group were treated with Chaishao Longmu decoction and those in the control group with eszopiclone for 4 weeks. The clinical efficacy， Pittsburgh sleep quality index （PSQI） score， TCM syndrome score and infrared thermal imaging characteristics ［temperature and temperature changes of frontal sleep line， frontal region， anterior trunk， Zhongwan （CV12）， conception vessel （CV）， left hypochondrium， right hypochondrium， dorsal trunk， and governor vessel （GV）］ of two groups were determined before and after treatment. ResultAfter treatment， the clinical response rate in treatment group was 91.67% （33/36）， which was higher than that （66.67%， 24/36） in the control group （Z=-2.617， P<0.01）. The treatment in both groups decreased the PSQI score and TCM syndrome score （P<0.01）， and the decreases were more significant in treatment group than in the control group （P<0.01）. After treatment， the total response rate of sleep line improvement in the treatment group was 86.11% （31/36）， which was higher than that （66.67%， 24/36） in the control group （Z=-2.591， P<0.05）. The frontal temperature of the two groups decreased （P<0.01） after treatment. Compared with those before treatment， the temperatures of anterior trunk， CV12， CV， left hypochondrium， right hypochondrium， dorsal trunk， and GV rose after treatment （P<0.01）. After treatment， the treatment group had lower frontal temperature and higher temperatures of anterior trunk， CV12， CV， left hypochondrium， right hypochondrium， dorsal trunk， and GV than the control group （P<0.01）. The treatment in the treatment group reduced the ∆T values of GV （P<0.01） and increased that of the CV12， CV， left hypochondrium， and right hypochondrium （P<0.05，P<0.01）. The treatment in the control group increased the ∆T value of CV12 （P<0.01）. After treatment， the treatment group had lower ∆T values of GV （P<0.01） and higher ∆T value of CV12， CV， left hypochondrium， and right hypochondrium （P<0.05， P<0.01） than the control group. Compared with that before treatment， the temperature difference between GV and CV in the two groups increased after treatment （P<0.01）. According to the infrared thermal image characteristics， the normal temperature difference between GV and CV was within the range of 0.5-1. The median value after treatment in the treatment group was 0.69 （0.52， 0.88）， which was within the normal range， indicating that the treatment group outperformed the control group. ConclusionChaishao Longmu decoction can alleviate short-term insomnia by soothing liver， invigorating spleen， harmonizing the middle energizer， and regulating GV and CV. With definite clinical effect， this decoction deserves promotion. Furthermore， the frontal temperature， sleep line， CV12， CV ∆T， and temperature difference between GV and CV revealed by the infrared thermal images could be used for the diagnosis and of insomnia with liver depression and spleen deficiency.

Biological Connotation of Pathogenesis of Ulcerative Colitis with Liver Depression and Spleen Deficiency Based on Bile Acid-intestinal Flora Axis / 中国实验方剂学杂志

Ulcerative colitis （UC） is one of the chronic refractory inflammatory bowel diseases characterized by abdominal pain， diarrhea， and mucus， pus and blood in the stool. In recent years， with changes in human life style and improvements of the diagnosis， the incidence and prevalence of UC have been increasing. The pathogenesis of UC is closely related to intestinal mucosal immune dysfunction， intestinal flora disturbance， and abnormal bile acid secretion. Patients with UC have abnormal bile acid secretion and intestinal flora imbalance. A large number of studies have found that abnormal bile acid secretion inhibits immune function， affects signal transduction， and destroys the intestinal mucosal barrier. Intestinal flora disturbance has an important impact on the occurrence and development of inflammation， immune homeostasis， and stress. Bile acids indirectly or directly affect the structure and function of intestinal flora， and at the same time， they produce secondary bile acids under the modification of intestinal flora， entering the liver through enterohepatic circulation. Therefore， the complex dialogue mechanism of bile acid-intestinal flora axis is closely related to the occurrence and development of UC. Based on the basic theory of traditional Chinese medicine（TCM） and clinical research， it is found that emotion is an important factor that induces this disease， spleen and stomach weakness is the root of the disease， and liver depression and spleen deficiency are the key pathogenesis of UC. Combined with modern medicine and molecular biology research， it is believed that abnormal secretion of bile acids is a microscopic manifestation of liver depression in TCM， and intestinal flora disturbance is the biological basis of spleen deficiency. In the pathogenesis of UC， the imbalanced bile acid-intestinal flora axis is consistent with the pathogenesis of liver depression and spleen in TCM. The exploration of the biological connotation of the pathogenesis of UC with liver depression and spleen deficiency from the perspective of bile acid-intestinal flora axis can better explain the scientific nature of its pathogenesis， which provides new clinical solutions and reliable references for studying the pathogenesis of UC with liver depression and spleen deficiency and finding representative prescriptions to prevent and treat this disease.

[Effect of Wei's triple nine needling on eye regulation in patients with presbyopia complicated with visual fatigue of liver depression and spleen deficiency].

OBJECTIVE: To compare the clinical efficacy between Wei's triple nine needling combined with esculin and digitalis glycosides eye drops and esculin and digitalis glycosides eye drops alone for presbyopia complicated with visual fatigue of liver depression and spleen deficiency. METHODS: Forty-six cases (92 eyes) with presbyopia complicated with visual fatigue of liver depression and spleen deficiency were randomly divided into an observation group (23 cases) and a control group (23 cases, 2 cases dropped off). The cases in the observation group were treated with Wei's triple nine needling and esculin and digitalis glycosides eye drops. The acupoints included Shangming (Extra), Chengqi (ST 1), Cuanzhu (BL 2) to Jingming (BL 1), Sizhukong (TE 23) to Taiyang (EX-HN 5), etc; the needling was given once every other day, three times a week, and the eye drops were given one drop each time, three times a day. The cases in the control group were only treated with the eye drops. Both groups were treated for 7 days as one course of treatment, and 2 courses of treatment were given. The visual fatigue core symptoms score, adjustment amplitude, adjustment lag and best average corrected visual acuity were observed in the two groups before treatment, 1 week and 2 weeks into treatment, respectively. RESULTS: Compared before treatment, the visual fatigue core symptoms scores in the two groups were decreased after 1-week and 2-week treatment (P<0.05); in the observation group, the adjustment amplitude was increased after 2-week treatment (P<0.05), while in the control group, the adjustment amplitude was increased after 1-week and 2-week treatment (P<0.05); in the observation group, the adjustment lag was decreased after 1-week and 2-week treatment (P<0.05). After 2-week treatment, the visual fatigue core symptoms score in the observation group was lower than that in the control group, and the adjustment amplitude was higher than that in the control group (P<0.05). There were no significant differences in adjustment lag and best average corrected visual acuity between the two groups after 1-week and 2-week treatment (P>0.05). CONCLUSION: Wei's triple nine needling combined with esculin and digitalis glycosides eye drops could improve the visual fatigue and eye regulation ability in patients with presbyopia complicated with visual fatigue of liver depression and spleen deficiency, and the effect is better than esculin and digitalis glycosides eye drops alone.

Expression of chitinase-3-like protein 1 in different TCM syndromes and its correlation with liver fibrosis.

BACKGROUND: We aimed to explore the expression of chitinase-3-like protein 1 (CHI3L1) in different traditional Chinese medicine (TCM) syndromes and its correlation with liver fibrosis. METHODS: A total of 95 patients with hepatitis B e antigen (HBeAg)-positive chronic hepatitis B who were diagnosed by pathological examination of liver biopsy specimens were selected as study subjects. According to different TCM syndrome types, 47 cases were divided into damp-heat moderate resistance type, and 48 cases were liver depression and spleen-deficiency type. Serum samples of the 2 groups were collected, then the levels of CHI3L1 were detected by enzyme-linked immunosorbent assay (ELISA). Hyaluronic acid (HA), laminin (LN), fibronectin (FN), collagen type IV (CIV), and N-terminal procollagen III propeptide (PIIINP) were measured by chemiluminescence. The levels of CHI3L1 was studied to explore the correlations between the 2 types of TCM syndromes and liver fibrosis. RESULTS: The serum CHI3L1 level of the 2 groups increased with increasing fibrosis stage and peaked at the S4 stage. The serum CHI3L1 levels of liver depression and spleen deficiency type patients in the S0-S1, S2-S3, and S4 stages were higher than those of patients with damp-heat moderate resistance type, and the difference was significant (P<0.01). The levels of HA, LN, FN, CIV, and PIIINP in the 2 groups increased with the increase of fibrosis stage and reached the highest level at the S4 stage. Compared to patients with damp-heat syndrome, the levels of HA, LN, FN, CIV, and PIIINP in patients with liver depression and spleen deficiency syndrome were lower, and the difference was statistically significant (P<0.05). There were positive correlations between CHI3L1 and HA, LN, FN, CIV, and PIIINP (all <0.05). CONCLUSIONS: The expression level of CHI3L1 in HBeAg-positive chronic hepatitis B patients was higher than expected. The levels of CHI3L1, HA, LN, FN, CIV, and PIIINP in liver depression and spleen deficiency type patients were lower than those in damp-heat moderate resistance type patients. These findings indicated that CHI3L1 is correlated with HA, LN, FN, CIV, PIIINP liver fibrosis indexes, and the syndrome of stagnation of spleen deficiency has a better correlation.

Effect of Wei's triple nine needling on eye regulation in patients with presbyopia complicated with visual fatigue of liver depression and spleen deficiency / 中国针灸

OBJECTIVE@#To compare the clinical efficacy between Wei's triple nine needling combined with esculin and digitalis glycosides eye drops and esculin and digitalis glycosides eye drops alone for presbyopia complicated with visual fatigue of liver depression and spleen deficiency.@*METHODS@#Forty-six cases (92 eyes) with presbyopia complicated with visual fatigue of liver depression and spleen deficiency were randomly divided into an observation group (23 cases) and a control group (23 cases, 2 cases dropped off). The cases in the observation group were treated with Wei's triple nine needling and esculin and digitalis glycosides eye drops. The acupoints included Shangming (Extra), Chengqi (ST 1), Cuanzhu (BL 2) to Jingming (BL 1), Sizhukong (TE 23) to Taiyang (EX-HN 5), etc; the needling was given once every other day, three times a week, and the eye drops were given one drop each time, three times a day. The cases in the control group were only treated with the eye drops. Both groups were treated for 7 days as one course of treatment, and 2 courses of treatment were given. The visual fatigue core symptoms score, adjustment amplitude, adjustment lag and best average corrected visual acuity were observed in the two groups before treatment, 1 week and 2 weeks into treatment, respectively.@*RESULTS@#Compared before treatment, the visual fatigue core symptoms scores in the two groups were decreased after 1-week and 2-week treatment (P<0.05); in the observation group, the adjustment amplitude was increased after 2-week treatment (P<0.05), while in the control group, the adjustment amplitude was increased after 1-week and 2-week treatment (P<0.05); in the observation group, the adjustment lag was decreased after 1-week and 2-week treatment (P<0.05). After 2-week treatment, the visual fatigue core symptoms score in the observation group was lower than that in the control group, and the adjustment amplitude was higher than that in the control group (P<0.05). There were no significant differences in adjustment lag and best average corrected visual acuity between the two groups after 1-week and 2-week treatment (P>0.05).@*CONCLUSION@#Wei's triple nine needling combined with esculin and digitalis glycosides eye drops could improve the visual fatigue and eye regulation ability in patients with presbyopia complicated with visual fatigue of liver depression and spleen deficiency, and the effect is better than esculin and digitalis glycosides eye drops alone.

Ganshuang Granule Alleviates Early Liver Fibrosis （S1 and S2） in Chronic Hepatitis B with Liver Depression Spleen Deficiency and Blood Stasis Syndrome / 中国实验方剂学杂志

ObjectiveTo explore the effect of Ganshuang granule on liver fibrosis （S1 and S2） in chronic hepatitis B （CHB） with liver depression spleen deficiency and blood stasis syndrome. MethodA total of 100 patients were classified into the control group （50 in total with 4 lost and 2 rejected， 44 finally included） and observation group （50 in total with 5 lost and 2 rejected， 43 finally included） with the random number table method. Both groups were given oral entecavir tablets （0.5 mg/time， once a day， 12 months）， and oral glutathione tablets was applied depending on the conditions of patients. In addition， the control group took the analog drug of Ganshuang granule （3 g/time， 3 times/day， 12 months） and the observation group received Ganshuang granules （3 g/time， 3 times/day， 12 months）， followed by histological examination of the liver by puncture biopsy. The two groups were compared in terms of inflammatory activity grade and fibrosis stage， as well as liver stiffness measure （LSM）， liver function， hepatitis B virus （HBV） DNA， liver depression and spleen deficiency syndrome score， aspartate aminotransferase （AST）-to-platelet ratio index （APRI）， and fibrosis index based on the four factors （FIB-4）. ResultAfter treatment， liver fibrosis in the observation group was milder than that in the control group （P<0.05） and the inflammatory activity grade in the observation group was lower than that in the control group （P<0.05）. The effective rate in down-regulating inflammatory activity grade in the observation group was 77.78% as compared with the 45.83% in the control group （χ2=5.546， P<0.05）. The effective rate in decreasing the fibrosis stage in the observation group was 59.26%， which was higher than that （16.67%） in the control group （χ2=9.669， P<0.01）. The LSM and score of the liver stagnation and spleen deficiency syndrome in the observation group were lower than those in the control group at the 6th months and 12th months of treatment （P<0.05，P<0.01）. The levels of alanine aminotransferase （ALT）， AST， and alkaline phosphatase （ALP） in the observation group were lower than those in the control group （P<0.01）. The APRI and FIB-4 in the observation group were lower than those in the control group （P<0.01）. ConclusionThe Ganshuang granule combined with entecavir can alleviate inflammation and liver fibrosis， delay and reverse liver fibrosis， protect liver， and improve the traditional Chinese medicine syndrome of liver fibrosis （S1 and S2） in CHB， which is worth of clinical use and further research.

[Mechanism of Xiaoyao San in treatment of depression,breast hyperplasia,and functional dyspepsia based on network pharmacology].

This study aimed to explore the mechanism of Xiaoyao San(XYS) in the treatment of three diseases of liver depression and spleen deficiency, ie, depression, breast hyperplasia, and functional dyspepsia, and to provide a theoretical basis for the interpretation of the scientific connotation of "treating different diseases with the same method" of traditional Chinese medicines. Traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP) was used to screen the active components of XYS which underwent principal component analysis(PCA) with the available drugs for these three diseases to determine the corresponding biological activities. The targets of XYS on depression, breast hyperplasia, and functional dyspepsia were obtained from GeneCards, TTD, CTD, and DrugBank databases. Cytoscape was used to plot the "individual herbal medicine-active components-potential targets" network. The resulting key targets were subjected to Kyoto encyclopedia of genes and genomes(KEGG) pathway analysis and gene ontology(GO) enrichment analysis. A total of 121 active components of XYS and 38 common targets in the treatment of depression, breast hyperplasia, and functional dyspepsia were collected. The key biological pathways were identified, including advanced glycation and products(AGEs)-receptor for advanced glycation and products(RAGE) signaling pathway in diabetic complications, HIF-1 signaling pathway, and cancer-related pathways. The key targets of XYS in the treatment of depression, breast hyperplasia, and functional dyspepsia included IL6, IL4, and TNF, and the key components were kaempferol, quercetin, aloe-emodin, etc. As revealed by the molecular docking, a strong affinity was observed between the key components and the key targets, which confirmed the results. The therapeutic efficacy of XYS in the treatment of diseases of liver depression and spleen deficiency was presumedly achieved by reducing the inflammatory reactions. The current findings are expected to provide novel research ideas and approaches to classify the scientific connotation of "treating different diseases with the same method" of Chinese medicines, as well as a theoretical basis for understanding the mechanism of XYS and exploring its clinical applications.

Clinical study on modified Sini Powder combined with acupoint application in the treatment of liver depression and spleen deficiency syndrome of diarrhea irritable bowel syndrome / 国际中医中药杂志

Objective:To evaluate the clinical efficacy of modified Sini Powder combined with acupoint application in the treatment of liver depression and spleen deficiency syndrome of diarrhea irritable bowel syndrome(IBS-D). Methods:A total of 104 IBS-D patients with liver depression and spleen deficiency syndrome in our hospital from October 2018 to December 2020 were randomly divided into two groups according to the random number table method, 52 in each group. Both groups took montmorillonite powder orally first. On this basis, the control group was treated with pivirium bromide tablets, and the study group was treated with Modified Sini Powder combined with acupoint application. Both groups were treated for 4 weeks. TCM symptom scores were performed before and after treatment. The levels of serum IL-6, IL-8 and TNF-α were detected by ELISA. The recovery time of stool characteristics, the disappearance time of abdominal pain and the recovery time of stool times were observed and recorded, and the clinical curative effect was evaluated. Results:The total effective rate was 92.3% (48/52) in the observation group and 75.0% (39/52) in the control group, there was significant difference between the two groups ( χ2=5.696, P=0.017). After treatment, the scores of abdominal pain, diarrhea, stool frequency, irritability, mental fatigue and hypochondriac pain in the observation group were significantly lower than those in the control group ( t values were 15.492, 16.827, 13.419, 10.831, 14.736,12.437, respectively, all Ps<0.001), and the levels of serum IL-6, IL-8 and TNF-α were significantly lower than those in the control group ( t values were 16.390, 21.528 and 18.734, respectively, all Ps<0.001). The recovery time of stool characteristics [(3.79 ± 0.63) d vs. (4.84 ± 0.79) d, t=7.493], the disappearance time of abdominal pain [(2.63 ± 0.32) d vs. (3.91 ± 0.37) d, t=18.869], and the recovery time of stool times [(3.26 ± 0.57) d vs. (4.19 ± 0.68) d, t=7.558] in the observation group were significantly shorter than those in the control group ( P<0.01). Conclusion:Modified Sini Powder combined with acupoint application can improve the clinical symptoms of IBS-D patients with liver depression and spleen deficiency syndrome, reduce the level of serum inflammatory cytokines and improve the curative effect.

Role of Counterevidence from Chinese Medicinal Prescription in Construction of Integrated Disease-syndrome Animal Model： Xiaoyaosan as Counterevidence for Depression of Liver Depression and Spleen Deficiency Syndrome / 中国实验方剂学杂志

Objective:To seed for stable time window of the integrated disease-syndrome animal model based on the counterevidence from Chinese medicinal prescriptions， and to verify syndrome stability and reliability. Method:A model of depression was established by exposing rats to chronic unpredictable mild stress （CUMS）， followed by body weight measurement， sugar water test， behavioral test， and brain 5-hydroxytryptamine（5-HT） detection. The identification of liver depression and spleen deficiency syndrome was conducted after the equivalent transformation of human clinical symptoms into macroscopic representations of rats. Based on the dynamically collected macroscopic representation scale， Xiaoyaosan was used to reversely verify the stability and reliability of the integrated disease-syndrome animal model of depression due to liver depression and spleen deficiency. Result:The sugar water consumption and the number of crossings and the total movement distance in the open field test of 16-week-old rats in the CUMS （eight weeks of CUMS） group were significantly lower than those in the normal group （<italic>P</italic><0.05）. According to the immunohistochemical results， the 5-HT content in hippocampal area CA2 of rats in the CUMS group was also significantly lowered as compared with that in the normal group（<italic>P</italic><0.05），which indicated that depression was successfully modeled. The liver depression and spleen deficiency syndrome was present in 14-week-old rats （six weeks after CUMS）of the CUMS group， and the number of rats experiencing the liver depression and spleen deficiency syndrome reached the peak in the 16th week （eight weeks after CUMS），accounting for 70% of the total number. Thereafter， the number decreased gradually. The syndrome scores of the 14-， 16-， 18-， 20-， and 22-week-old rats in the Xiaoyaosan group were reduced by 66.6%， 70.7%， 54.8%， 50.4%， and 44.8%， which were graded as effective， marked effective， effective， effective， and effective， respectively. Conclusion:The age of 14-16 weeks（six to eight weeks after CUMS） is considered the stable and reliable time window for depression due to liver depression and spleen deficiency.

Mechanism of Xiaoyao San in treatment of depression,breast hyperplasia,and functional dyspepsia based on network pharmacology / 中国中药杂志

This study aimed to explore the mechanism of Xiaoyao San(XYS) in the treatment of three diseases of liver depression and spleen deficiency, ie, depression, breast hyperplasia, and functional dyspepsia, and to provide a theoretical basis for the interpretation of the scientific connotation of "treating different diseases with the same method" of traditional Chinese medicines. Traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP) was used to screen the active components of XYS which underwent principal component analysis(PCA) with the available drugs for these three diseases to determine the corresponding biological activities. The targets of XYS on depression, breast hyperplasia, and functional dyspepsia were obtained from GeneCards, TTD, CTD, and DrugBank databases. Cytoscape was used to plot the "individual herbal medicine-active components-potential targets" network. The resulting key targets were subjected to Kyoto encyclopedia of genes and genomes(KEGG) pathway analysis and gene ontology(GO) enrichment analysis. A total of 121 active components of XYS and 38 common targets in the treatment of depression, breast hyperplasia, and functional dyspepsia were collected. The key biological pathways were identified, including advanced glycation and products(AGEs)-receptor for advanced glycation and products(RAGE) signaling pathway in diabetic complications, HIF-1 signaling pathway, and cancer-related pathways. The key targets of XYS in the treatment of depression, breast hyperplasia, and functional dyspepsia included IL6, IL4, and TNF, and the key components were kaempferol, quercetin, aloe-emodin, etc. As revealed by the molecular docking, a strong affinity was observed between the key components and the key targets, which confirmed the results. The therapeutic efficacy of XYS in the treatment of diseases of liver depression and spleen deficiency was presumedly achieved by reducing the inflammatory reactions. The current findings are expected to provide novel research ideas and approaches to classify the scientific connotation of "treating different diseases with the same method" of Chinese medicines, as well as a theoretical basis for understanding the mechanism of XYS and exploring its clinical applications.

The characteristics of intestinal flora of IBS-D with different syndromes.

OBJECTIVE: To study the distribution of fecal microbiota in diarrhea-predominant irritable bowel syndrome (IBS-D) patients of spleen-kidney-yang deficiency and liver depression and spleen deficiency, to make an objective foundation for dialectics of different type of IBS-D. And to provide the clinical doctors an experimental basis for medication by regulating dysbacteriosis. METHODS: We collected feces from the control group, spleen-kidney-yang deficiency IBS-D group, and liver-depression and spleen-deficiency IBS-D group. After the extraction of fecal DNA, global DNA was isolated from every sample, and 16S ribosomal RNA was sequenced, and then we analyzed the results for bacteria such as Alpha diversity, community composition, LEfSe, and partial least squares discriminant analysis. RESULTS: We compared the changes among the fecal bacteria in the intestine of the IBS-D patients and healthy controls and found the specificity of spleen-kidney-yang deficiency syndrome and liver-depression and spleen-deficiency syndrome. The control group has the highest flora diversity (control group > liver-depression and spleen-deficiency > spleen-kidney-yang deficiency group). The control group, spleen-kidney-yang deficiency group, and liver-depression and spleen-deficiency group are different in phylum (Actinobacteria, Fusobacteria), class (Actinobacteria, Fusobacteria), order (Enterobacteriales, Bifidobacteriales, Fusobacteriales), and family (Bifidobacteriaceae, Ruminococcaceae, Enterobacteriaceae, Acidaminococcaceae, Veillonellaceae, Fusobacteriaceae). Bifidobacteriaceae and Ruminococcaceae in the control group, Enterobacteriales, Fusobacteriales, Acidaminococcaceae, and Phascolarctobacterium in the spleen-kidney-yang deficiency group, and streptococcus are the specific bacteria in the liver-depression and spleen-deficiency group. Intestinal flora disturbance is closely related to IBS-D. CONCLUSIONS: There is a correlation between traditional Chinese medicine syndrome type and intestinal flora. The control group, the spleen-kidney-yang deficiency group, and the liver-depression and spleen-deficiency group have specific bacteria.

Effect and Mechanism of Modified Banxia Xiexintang on Depression During Perimenopause / 中国实验方剂学杂志

Objective::To observe the effect of modified Banxia Xiexintang on depression during perimenopause, in order to study itseffecton 5-hydroxytryptamine (5-HT) and proinflammatory factors. Method::One hundred and thirty-nine patients were randomly divided into control group (69 cases) and observation group (70 cases) by random number table.Patients in control group got tibolone tablets, 2.5 mg/time, 1 time/day, and paroxetine hydrochloride tablets, 20 mg/time, 1 time/day.In addition to the therapy in control group, patients in observation group were added with modified Banxia Xiexintang, 1 dose/day.The course of treatment was 8 weeks.And before and after treatment, Hamilton depression scale for-17 items (HAMD-17), Zung's self-rating depression scale (SDS), hamilton anxiety scale (HAMA), improvement Kupperman(KI), liver depression and spleen deficiency syndrome, menopause-specific quality of life questionnaire (MENQOL) and treatment emergent symptom scale (TESS) were scored, and levels of 5-HT, rain-derived neurotrophic factor (BDNF), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were detected. Result::After treatment, scores of HAMD-17 and SDS in observation group were lower than those in control group (P<0.01). And the effect on trea depression in observation group was better than that in control group (Z=2.074, P<0.05). The degree of depression in observation group was ligher than that in control group (Z=2.157, P<0.05). And scores of HAMA, KI and liver depression and spleen deficiency syndrome were lower than those in control group (P<0.01). The severity of perimenopausal syndrome was ligher than that in control group (Z=2.046, P<0.05). And scores of vasomotor symptoms and psychological symptoms of MENQOL scale and the total scores were lower than those in control group (P<0.05). Levels of 5-HT and BDNF were higher than those in control group (P<0.01), while levels of IL-1β, TNF-α and TESS were lower than those in control group (P<0.01). Conclusion::In addition to theroutine western medicine, modified Banxia Xiexintang can alleviate the severity of depression, release the symptoms of depression, anxiety and perimenopausal syudrome(PMS), improve the quality of life, inhibit pro-inflammatory factors, and enhance the expressions of 5-HT and BDNF, with no adverse event.

Discussion of Correlation Between Chronic Psychological Stress Induced Liver Depression and Spleen Deficiency Syndrome and Microbiota-gut-brain Axis and Study on Intervention Mechanism of Xiaoyaosan / 中国实验方剂学杂志

Psychological stress （pressure） has gradually become an important factor affecting human physical and mental health， and is one of the important factors in the progression of clinical chronic refractory diseases. Psychological stress response can be attributed to the category of emotional illness in traditional Chinese medicine （TCM）. In the theory of TCM， the liver controls dispersion and regulates mental activity. Relevant scholars believe that the liver is the core of psychological stress response in the TCM theory. When being stimulated by chronic or repeated psychological stress， the body can gradually change from the initial depression and anxiety-rela behaviors to the gastrointestinal dysfunctions， which is similar to the formation process of the syndrome of liver depression and spleen deficiency with liver-spleen disharmony. The syndrome of liver depression and spleen deficiency is also the common syndrome of TCM for psychological stress-related diseases. With the effect in soothing liver and invigorating spleen， Xiaoyaosan has been regarded as the classic formula for anti-stress. Modern studies have showed that gut microbiota not only get involved in the movement， structure and function of the digestive tract， but also affect the brain function and behavior of the host through the gut-brain axis. Therefore， the microbiota-gut-brain axis has become an important part of psychological stress to trigger the body's gastrointestinal symptoms and abnormal brain behaviors. Focusing on psychological stress， the authors explored the correlation between the syndrome of liver stagnation and spleen deficiency and the microbiota-gut-brain axis， and the intervention mechanism of Xiaoyaosan， so as to enrich the scientific connotation of the syndrome of liver stagnation and spleen deficiency.

Effects of RuanJianXiaoYing Granule on expressions of Treg and Th17 cytokines in Hashimoto thyroiditis model rats with liver depression and spleen deficiency / 吉林大学学报(医学版)

Objective: To observe the effects of RuanJianXiaoYing Granule on the expressions of specific transcription factors and cytokines of Treg and Th17 in the Hashimoto thyroiditis (HT) rats with liver depression and spleen deficiency, and to investigate the therapeutic effect of RuanJianXiaoYing Granule on HT and its immunological mechanism. Methods: A total of 48 SD rats were randomly divided into normal group (12 rats) and modeling group (36 rats). The HT model was established by subcutaneous injection of excessive iodine drinking water and thyroglobulin, and then combined with chronic restraint stress, excessive fatigue and dietary disorders to prepare the rat models with liver depression and spleen deficiency. After modeling, the rats were randomly divided into model group (n=12), Tripterygium wilfordii group (n=12), and RuanJianXiaoYing group (n=12), and the rats were treated for 8 weeks. In addition to the dead rats and unqualified specimens during the experiment, the test results of 10 rats in each group were retained. The general situation of the rats in various groups was observed; the abdominal aortic blood and thyroid gland tissue of the rats in various groups were collected after the last administration; the levels of thyroglobulin antibody (TGAb), thyroid peroxidase antibody (TPOAb), free triiodothyronine (FT3), free thyroxine (FT4), thyrotropin-releasing hormone (TRH) and thyrotropin (TSH) in serum and the levels of IL-17, IL-23 and IL-10 were detected by ELISA method; the pathomorphology of thyroid gland tissue of the rats in various groups were detected by HE staining; the expression levels of Foxp3 and RORyt proteins in thyroid gland tissue of the rats in various groups were detected by Western blotting method. Results: Compared with normal group, the levels of serum TGAb and TPOAb of the rats in model group, Tripterygium wilfordii group and RuanJianXiaoYing - group were increased (P0. 05); the level of serum IL-10 was increased (P<0. 05), and the levels of serum IL-17 and IL-23 were decreased (P<0. 05). The follicular cavity of thyroid gland in normal group had regular morphology, complete structure and no lymphocyte infiltration. In model group, the follicular cavity of thyroid gland was enlarged, the follicular structure was destroyed, and a large number of lymphocyte infiltration was observed in and around the follicular cavity. In Tripterygium wilfordii group, the follicular cavity of thyroid gland was enlarged, part of the follicular structure was destroyed, and there was a small amount of lymphocyte infiltration in and around the follicular cavity, which was less than that in model group. The morphological changes of thyroid gland in RuanJianXiaoYing group were similar to those in Tripterygium wilfordii group Conclusion: RuanJianXiaoYing Granule has the therapeutic effect in the HT rats with liver depression and spleen deficiency by regulating the expressions of the specific transcription factors Foxp3 and RORγt protein and related cytokines of Treg and Th17.

Efficacy of Jianpi Shugan Fomula on Intestinal Flora of Patients with Diarrhea Predominant Irritable Bowel Syndrome and Its Effect on Intestinal Flora / 中国实验方剂学杂志

Objective:To observe the effect of Jianpi Shugan decoction combined with acupuncture on the clinical symptoms of diarrhea-predominant irritable bowel syndrome (IBS-D) with liver depression and spleen deficiency and the intestinal flora. Method:Seventy patients with IBS-D with liver depression and spleen deficiency were randomly divided into two groups by the random number table. The treatment group was given Jianpi Shugan decoction combined with acupuncture for 4 weeks, the control group was treated with Pivavironium bromide for 4 weeks, and 30 healthy people were used as healthy control. The total effective rate, IBS bowel symptom severity scale (IBS-BSS), IBS quality of life questionnaire (IBS-QOL) and traditional Chinese medicine pattern curative effect scoring system (TCM-PES) were evaluated. The counts of Bacillus bifidus, B.acidi lactici, Enterobacteria, and Bacteroides in feces and the colonization resistance (CR) were observed by Real-time PCR. Result:The IBS-SSS scale showed that the TCM treatment group could reduce the scores at the 4th, 8th, and 12th weeks (Pth and 8th weeks (Pth week scores of TCM treatment group were better than that of control group (PPth, 8th, and 12th weeks, and the control group increased the scores at the 4th week (PPB.bifidus and B.acidi lactici increased after 4 weeks of TCM treatment, and the count of Enterobacteria decreased (PB. bifidus increased, while Enterobacteria decreased in the TCM treatment group (PBacteroides after treatment in each group. Conclusion:Jianpi Shugan decoction combined with acupuncture has a reliable curative effect on IBS-D patients with liver depression and spleen deficiency. The mechanism may be related to the regulation of intestinal flora imbalance.

Clinical Observation of Jiawei Xiaoyaosan in Treating Liver Depression and Spleen Deficiency Type 2 Diabetes with Emotional Disorder / 中国实验方剂学杂志

Objective:To observe the clinical efficacy of Jiawei Xiaoyaosan for patients with anxiety and depression in type 2 diabetes with liver depression and spleen deficiency. Method:The 76 eligible patients with anxiety and depression in type 2 diabetes with liver depression and spleen deficiency were randomly divided into two groups:control group (38 cases) and treatment group (38 cases). In control group, the basic medicines metformin sustained-release tablets combined with deanxit were given; based on the treatment in control group, the patients in treatment additionally received Jiawei Xiaoyaosan. The changes of fasting plasma glucose (FPG), 2 hours postprandial blood glucose (2 hPG), haemoglobin A1c (HbA1c), fasting insulin (FINS) and serum insulin levels 30 minutes after glucose intake (30 min INS), Hamilton depression scale (HAMD), Hamilton anxiety scale (HAMA) and so on were observed and compared. Result:As compared with those before treatment, the levels of FPG, 2 hPG and HbA1c were decreased significantly (PPPPPPPPConclusion:Jiawei Xiaoyaosan combined with basic medication can significantly reduce depression and anxiety in patients with type 2 diabetes mellitus of liver depression and spleen deficiency, reduce blood glucose, glycosylated hemoglobin, decrease fasting insulin and increase 30 min INS level. The effect may be related to the improvement of anxiety, depression and other adverse emotions in these patients.

[Methods to establishment of serum induced syndrome of liver-depression and spleen-deficiency cell model].

To explore the method of establishing a cell model of traditional Chinese medicine (TCM) syndromes, HepG2 cells were induced by human serum of liver-depression and spleen-deficiency syndrome(LDSDS) to establish a cell model of LDSDS in this research. The concentration of cells, the content of human serum in culture medium and the growth characteristics of model-cell (cell growth curve, the survival rate and apparent morphology were investigated by MTT assay and microscopy. Evaluation of syndrome cell model: metabolomics was used to analyze the human serum of normal individuals and patients with LDSDS, and cell models induced by these serums, respectively. We obtained the difference metabolites from serums and cell models of LDSDS, respectively; then compared the biomarkers from two metabolomics and their metabolic pathways, to verify that the reliability and applicability of the model. Metabolomics data were collected by UPLC-Q-TOF-MS, and then all data were analyzed by multivariate statistical (PCA，OPLS-DA). The results showed that, model cells have the characteristics of normal growth, slow proliferation and stable morphological structure inducted by 10% serum of LDSD in 24-72 h. There were the same 19 difference metabolites which from the human serum of normal individuals and patients with LDSDS, and cell models induced by these serums; including 9 metabolic pathways that play an important role in maintaining normal physiological activities of the human body, such as lipids, amino acids, nucleotides, and energy metabolism etc. It was shown that the established syndrome cell model can reflect the biological basis of LDSDS to some extent. This research provides a reference method for the establishment of TCM syndrome cell model.