Syndromes and Mechanisms of Depression Induced by Second Hit in Mice / 中国实验方剂学杂志

ObjectiveTo explore the syndromes and mechanisms of depression induced by maternal separation （MS） combined with chronic restraint stress （RS） in mice. MethodOn postnatal day 0 （PD0）， the offspring mice were randomized into a blank group （NC） and a modeling group. The mouse model of depression was established by MS+RS for 21 days. After removal of female mice on PD21， the modeled mice were randomized into model， Wenyang， Jieyu， Wenyang Jieyu， and fluoxetine groups， with 15 mice in each group. The sucrose preference， tail suspension， and open field tests were carried out to evaluate the anxiety and depression-like behavior in mice. Enzyme-linked immunosorbent assay was used to measure the adrenocorticotrophic hormone （ACTH） and corticosterone （CORT） levels in mouse plasma. High performance liquid chromatography-electrochemical detector was used to determine the content of monoamine neurotransmitters in the hippocampus. Real-time fluorescence quantitative polymerase chain reaction was employed to determine the mRNA levels of genes in the 5-hydroxytryptamine （5-HT） system， hypothalamic-pituitary-adrenal （HPA） axis， and brain-derived neurotrophic factor （BDNF） signaling pathway in the hippocampus. Immunohistochemistry was employed to determine the expression levels of proteins in the 5-HT system and HPA axis in the hippocampus. The Simple Western system was used to determine the protein levels of BDNF and tyrosine kinase receptor B （TrkB） in the hippocampus. ResultCompared with the NC group， the model group exhibited depression-like behavior， which was significantly relieved by Wenyang Jieyu prescription and fluoxetine. Compared with the NC group， the model group showed elevated levels of CORT and ACTH in the plasma （P<0.01）， which， however， were lowered by Wenyang Jieyu prescription and fluoxetine （P<0.05， P<0.01）. Compared with the NC group， the model group showed inhibited expression of neurotransmitters in the hippocampus （P<0.05， P<0.01）， while Wenyang Jieyu prescription and fluoxetine restored the expression of neurotransmitters （P<0.05， P<0.01）. Compared with NC group， the model group showed inhibition of the 5-HTergic nerve and abnormal activation of the HPA axis， and Wenyang Jieyu prescription and fluoxetine regulated the abnormal state of the 5-HTergic nerve and HPA axis. Compared with NC group， the modeling down-regulated the mRNA and protein levels of BDNF and TrkB in the hippocampus （P<0.05， P<0.01）， which， however， were recovered in Wenyang， Jieyu， Wenyang Jieyu， and fluoxetine groups （P<0.05， P<0.01）. ConclusionThe mouse model of depression induced by MS+RS may present the syndrome of Yang deficiency and liver depression. Wenyang Jieyu prescription may increase the content of hippocampal neurotransmitters by regulating the 5-HT system and the BDNF signaling pathway mediated by the HPA axis， thereby alleviating depression-like behavior in mice.

Clinical study on Xiaosheng Powder ultrasonic nebulization combined with meibomian gland massage for the patients with meibomian gland dysfunction dry eye and pattern of liver depression and Yin deficiency / 国际中医中药杂志

Objective:To observe the clinical efficacy of Xiaosheng Powder ultrasonic nebulization combined with meibomian gland massage in treating liver depression and Yin deficiency pattern of meibomian gland dysfunction dry eye.Methods:Randomized controlled trial. From March 2021 to September 2021, a total of 50 patients (100 eyes) diagnosed with meibomian gland dysfunction dry eye of the liver depression and Yin deficiency pattern at the Ophthalmology Hospital of China Academy of Chinese Medical Sciences were selected. They were randomly divided into two groups using a random number table, with 25 patients (50 eyes) in each group. The control group received 0.1% sodium hyaluronate eye drops combined with meibomian gland massage, while the treatment group received Xiaosheng Powder nebulization combined with meibomian gland massage. Both groups were treated for 4 weeks. Before and after treatment, the tear film break-up time (BUT), Schirmer I test (SIT), corneal fluorescein staining (FL) scores, TCM symptom scores, and meibomian gland function scores were observed and evaluated. Adverse reactions were recorded.Results:After treatment, the treatment group exhibited statistically significant improvements compared to the control group in the following aspects: BUT was significantly longer in the treatment group (t=8.76, P<0.01); SIT values were significantly higher in the treatment group ( t=6.18, P<0.01); FL scores, TCM symptom scores and meibomian gland function scores were significantly lower than in the treatment group ( t values were 2.19, 5.36 and 12.09, P<0.01). The total effective rate in the treatment group was 96.0% (24/25), while in the control group, it was 68.0% (17/25), and the difference between the two groups was statistically significant ( χ2=4.88, P=0.027). Conclusion:The combination of Xiaosheng Powder ultrasonic nebulization and meibomian gland massage is effective in improving the clinical symptoms of patients with meibomian gland dysfunction dry eye of the liver depression and Yin deficiency pattern. Furthermore, this approach is considered safe and efficient.

Observation on the Clinical Efficacy of Modified Tongxie Yaofangin Treating Diarrhea-predominant Irritable Bowel Syndrome with Liver-Depression and Spleen-Deficiency Syndrome / 广州中医药大学学报

Objective To observe the clinical efficacy of Modified Tongxie Yaofang(Important Formula for Relieving Diarrhea with Pain)in the treatment of patients with diarrhea-predominant irritable bowel syndrome(IBS-D)with liver-depression and spleen-deficiency syndrome.Methods Sixty patients with IBS-D of liver depression and spleen deficiency type were randomly divided into treatment group and control group,30 patients in each group.The patients in the treatment group were given modified Tongxie Yaofang,and the patients in the control group were given Pivacurium Bromide Tablets(Dicetel).Both groups were treated for a 4-week course of treatment.The changes of traditional Chinese medicine(TCM)syndrome scores,irritable bowel syndrome quality of life(IBS-QOL)scores,Hamilton Anxiety Scale(HAMA)scores of the two groups were observed before and after the treatment.Moreover,the efficacy for overall symptomatic improvement in the two groups was evaluated according to the grading of irritable bowel syndrome symptom severity score(IBS-SSS),and the efficacy for improving single symptom such as abdominal pain and diarrhea as well as the relapse after 4 weeks of drug cessation in the two groups were compared.Results(1)After 4 weeks of treatment,the total effective rate for overall symptomatic improvement in the treatment group was 83.33%(25/30),and that in the control group was 43.33%(13/30).The intergroup comparison(tested by chi-square test)showed that the efficacy for overall symptomatic improvement in the treatment group was significantly superior to that in the control group(P＜0.01).(2)After 4 weeks of treatment,the total effective rate for improving single symptom of abdominal pain and diarrhea in the treatment group was 80.00%(24/30),90.00%(27/30),and that in the control group was 43.33%(13/30),46.67%(14/30),respectively.The intergroup comparison(tested by chi-square test)showed that the efficacy for improving single symptom of abdominal pain and diarrhea in the treatment group was significantly superior to that in the control group(P＜0.01).(3)After 4 weeks of treatment,the TCM syndrome scores,IBS-QOL scores,and HAMA scores of patients in both groups were significantly decreased compared with those before treatment(P＜0.05 or P＜0.01),and the decrease in the treatment group was significantly superior to that in the control group(P＜0.01).(4)After 4 weeks of drug withdrawal,the recurrence rate of the treatment group was 24.00%(6/25),which was significantly lower than that of the control group(61.54%,8/13),and the difference was statistically significant between the two groups(P＜0.05).Conclusion Modified Tongxie Yaofang exerts certain effect in treating patients with IBS-D of liver depression and spleen deficiency type,and the decoction is effective on decreasing the scores of symptoms of abdominal pain and diarrhea as well as TCM syndrome scores,improving the quality of life of the patients,and alleviating the anxiety status of the patients.

Clinical Study on Therapy of Soothing Liver and Strengthening Spleen in the Treatment of Active Thyroid-Associated Ophthalmopathy Complicated with Dry Eye / 广州中医药大学学报

Objective To investigate the efficacy of the therapy of soothing liver and strengthening spleen(shortened as Shugan Jianpi therapy)in the treatment of active thyroid-associated ophthalmopathy(TAO)complicated with dry eye,and to provide a reference basis for clinical treatment.Methods A total of 108 patients with active TAO complicated with dry eye of liver depression and qi stagnation type were randomly divided into observation group and control group,54 patients in each group.Both groups were given conventional treatment for intervention of Graves'disease,and additionally the control group was given hormone shock therapy by intravenous injection of Methylprednisolone Sodium Succinate,and the observation group was treated with Chinese medicine prescription for soothing liver and strengthening spleen orally and intravenous injection of Methylprednisolone Sodium Succinate.The treatment period lasted for 12 weeks,and then the patients were followed up till to the 6th month.The changes of clinical activity score(CAS),proptosis,ocular surface disease index(OSDI),corneal fluorescein staining(FL),Schirmer I test(SIT)and tear film break-up time(BUT)in the two groups were observed before and after the treatment.After treatment,the clinical efficacy of the two groups was evaluated.Results(1)After 6 months of treatment,the total effective rate in the observation group was 94.44%(51/54)and that in the control group was 74.07%(40/54),and the intergroup comparison(tested by chi-square test)showed that the efficacy of the observation group was significantly superior to that of the control group(P＜0.05).(2)After treatment,the CAS,OSDI score and proptosis of the patients in the two groups were all lower than those before treatment(P＜0.01),and the reduction in the observation group was significantly superior to that in the control group(P＜0.01).(3)After treatment,the indicators of tear secretion function such as SIT,FL score and BUT of patients in the two groups were improved compared with those before treatment(P＜0.01),and the improvement in the observation group was significantly superior to that in the control group,the differences being all statistically significant(P＜0.01).Conclusion Shugan Jianpi therapy exerts certain clinical efficacy in treating patients with active TAO complicated with dry eye of liver depression and qi stagnation type,which can effectively relieve the proptosis,prolong the tear film break-up time,promote the secretion of tears and the repair of corneal epithelium,improve the visual function,and enhance the quality of life of the patients.

Combined Therapies of Traditional Chinese Medicine Treat Low-prognosis Patients Undergoing IVF-ET Again / 中国实验方剂学杂志

ObjectiveTo objectively evaluate the clinical efficacy of multiple therapies of traditional Chinese medicine （TCM） in low-prognosis patients who received antagonist protocol for in vitro fertilization and embryo transfer （IVF-ET） again. MethodA total of 128 patients with kidney Yin deficiency， liver depression， and blood stasis who planned to receive antagonist protocol for IVF-ET in the West China Second Hospital of Sichuan University were enrolled and assigned into two groups by random number table method. The observation group （64 casces） was treated by oral administration of Chinese medicine decoction + enema of kidney-tonifying and blood-activating method + auricular point sticking + oral administration of dehydroepiandrosterone （DHEA）， while the control group （64 casces） was treated by only oral administration of DHEA. After treatment for three menstrual cycles， both groups received the antagonist protocol for IVF-ET. The TCM syndrome scores， basic sex hormone levels， antral follicle count （AFC）， the usage of gonadotropin （Gn）， endometrial receptivity indicators， embryo quality indicators， and pregnancy outcomes were compared between the two groups. ResultAfter treatment， the observation group showed decreased follicle-stimulating hormone （FSH）/luteinizing hormone （LH） ratio， lowered level of estradiol （E2）， increased AFC， decreased amount and days of Gn usage， improved endometrial receptivity indicators （endometrial thickness on trigger and ET days， proportion of endometrial type A in endometrial types and the level of E2 on trigger day） and embryo quality indicators （the rates of mature follicles， fertilization， normal fertilization， and premium embryos）， and decreased TCM syndrome scores （P<0.05， P<0.01）. Moreover， the observation group had lower FSH/LH ratio， E2 level， and amount of Gn usage， higher AFC， poorer endometrial receptivity and embryo quality indicators， and lower TCM syndrome scores than the control group after treatment （P<0.05， P<0.01）. In addition， except for 3 cases of natural pregnancy， the observation group outperformed the control group in terms of improving the clinical pregnancy rates during initiation cycle and transplantation cycle and clinical pregnancy rate and decreasing biochemical pregnancy rate and early abortion rate （P<0.05）. ConclusionCombined therapies of TCM can alleviate the clinical symptoms， reduce TCM syndrome scores， reduce the Gn usage amount， improve the number and quality of embryos and endometrial receptivity， and coordinate the synchronous development of endometrium and embryo. In this way， they can increase the clinical pregnancy rate and reduce biochemical pregnancy rate and early abortion rate in the low prognosis patients with kidney yin deficiency， liver depression， and blood stasis who are undergoing IVF-ET again.

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.

Sishenwan Combined with Tongxie Yaofang Treats Ulcerative Colitis with Spleen-kidney Yang Deficiency and Liver Depression / 中国实验方剂学杂志

ObjectiveTo induce the rat model of ulcerative colitis （UC） with spleen-kidney Yang deficiency and liver depression， and explore the efficacy and mechanism of Sishenwan combined with Tongxie Yaofang （SSW&TXYF） based on the therapeutic principles of tonifying spleen， soothing liver， warming kidney， and astringing intestine. MethodSixty male SD rats were randomized into normal， model， mesalazine， and high-， medium-， and low-dose SSW&TXYF groups. The rats in other groups except the normal group were administrated with Sennae Folium decoction and hydrocortisone and received tail clamping for 14 days. On day 14， rats received enema with TNBS-ethanol solution to induce UC. The rats were administrated with corresponding drugs from day 15 of modeling， and the body weight and mental state were observed and recorded. The sucrose preference test was performed from day 25. On day 28， the rectal temperature was measured， and the rats were administrated with 3% D-xylose solution at a dose of 10 mL·kg-1 by gavage. Blood was sampled 1 h later， from which the serum was collected for measurement of the D-xylose content. The serum， hippocampus， and colorectum samples of rats were collected on day 29. The levels of gastrin （GAS）， adrenocorticotropic hormone （ACTH）， corticosterone （CORT）， cyclic adenosine monophosphate （cAMP）， cyclic guanosine monophosphate （cGMP）， interleukin （IL）-4， tumor necrosis factor （TNF）-α， and interferon （IFN）-γ in the serum and 5-hydroxytryptamine （5-HT） in the hippocampus were determined by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining was employed to reveal the colonic lesions. The mRNA and protein levels of p38 mitogen-activated protein kinase （MAPK）， extracellular signal-regulated kinase （ERK）， and c-Jun N-terminal kinase （JNK） in the colon tissue were determined by Real-time PCR and Western blot， respectively. ResultCompared with the normal group， the model group showed decreased body weight， anal temperature， and D-xylose content in the serum and increased GAS content （P<0.01）. The modeling led to cAMP/cGMP unbalance and decreased the ACTH and CORT content in the serum （P<0.01）， the preference for sucrose water， and the 5-HT content in the hippocampus （P<0.01）. Moreover， it shortened the colorectal length and caused massive infiltration of inflammatory cells and severe structural damage in the colon tissue. High， medium， and low doses of SSW&TXYF improved above indicators （P<0.05， P<0.01）， reduced inflammatory infiltration， and repaired the pathological damage of the tissue. Compared with the normal group， the model group showed lowered IL-4 level （P<0.01） and elevated TNF-α and IFN-γ levels （P<0.05， P<0.01） in the serum， as well as up-regulated expression of p38 MAPK， ERK， and JNK （P<0.05， P<0.01）. Compared with the model group， SSW&TXYF elevated the IL-4 level （P<0.01）， lowered the TNF-α and IFN-γ levels （P<0.05， P<0.01）， and down-regulated the mRNA and protein levels of p38 MAPK， ERK， and JNK （P<0.05， P<0.01）. ConclusionA rat model of UC with spleen-kidney Yang deficiency and liver depression was successfully established. SSW&TXYF can significantly mitigate this syndrome by reducing the inflammatory response in the colon and inhibiting the MAPK pathway.

Effect of processing with vinegar on efficacy of Curcuma longa in treatment of dysmenorrhea in rats with syndrome of liver depression and Qi stagnation / 中国中药杂志

This study compared the effects of Curcuma longa before and after processing with vinegar on the rat model of dysmenorrhea with the syndrome of liver depression and Qi stagnation to reveal the mechanism of vinegar processing in improving the role of C. longa in soothing liver and relieving pain. The rat model of dysmenorrhea with the syndrome of liver depression and Qi stagnation was established according to the Preparation of the Animal Model of Dysmenorrhea(Draft) and the chronic unpredictable stress me-thod. The changes in the body weight, organ indexes, writhing latency, writhing score, and serum levels of six liver function indicators, sex hormones, pain factors, and blood rheological indicators were measured to evaluate the efficacy of C. longa processed with vinegar or not in treating dysmenorrhea in the rats with syndrome of liver depression and qi stagnation. Compared with the model group, the C. longa group(processed with vinegar or not) showed slow weight loss, increase in writhing latency, and decrease in writhing response(P<0.05). The inhibition rates on writhing in raw C. longa, vinegar-processed C. longa, and positive groups were 33.780%, 64.611%, and 62.466%, respectively. The significantly higher inhibition rate of the vinegar processing group indicated that vinegar-processed C. longa demonstrated more significant therapeutic effect. The vinegar-processed C. longa group showed lower levels of alanine aminotransferase(ALT), alkaline phosphatase(ALP), aspartate aminotransferase(AST), direct bilirubin(DBIL), and total bilirubin(TBIL) and higher level of albumin(ALB)(P<0.05), which indicated that vinegar processing enhanced the therapeutic effect of C. longa on liver injury. The serum levels of estradiol(E_2) and oxytocin(OT) were lower in the vinegar-processed C. longa group(P<0.05), indicating that the vinegar-processed C. longa could regulate the sex hormone levels, reduce the activity of uterine smooth muscle and contraction of uterus, and alleviate the symptoms of dysmenorrhea in rats. Moreover, the vinegar-processed C. longa group showed lower interleukin-6(IL-6) and arginine vasopressin(AVP) levels and higher beta-endorphin(β-EP) level(P<0.05), which indicated that vinegar-processed C. longa regulated the levels of pain factors to exert the pain-relieving effect. Drug intervention decreased the whole blood viscosity low-cut, medium-cut and high-cut values, plasma viscosity, whole blood reduction viscosity low-cut and high-cut values, erythrocyte cumulative pressure, and equation K value of erythrocyte sedimentation rate(P<0.05), and the vinegar-processed C. longa group outperformed other groups. This result indicated that vinegar processing enhanced the function of C. longa in improving the local blood rheology. C. longa processed with vinegar can enter the liver to relieve the da-mage to the heart, liver, kidney, and uterus, repair the liver function, and recover the sex hormone levels and immune function by regulating the levels of sex hormones and pain factors and improving the blood rheology. It activates the pain-relieving mechanism to relieve the pain, protect the liver, and fight inflammation, which is consistent with the theory that vinegar processing facilitates C. longa entering the liver to sooth liver and relieve pain.

Correlation between Clinical Manifestations and Gut Microbiota of URSA Patients with the Syndrome of Liver Depression and Kidney Deficiency / 世界科学技术-中医药现代化

Objective Comparing the differences of gut microbiota between healthy women and URSA patients with the syndrome of liver depression and kidney deficiency,and the relationship between gut microbiota and age,abortion times,TCM syndrome score,and DASS-42 in URSA patients.Exploring the relationship between gut microbiota and clinical symptoms.Methods The observation group was 28 unpregnant URSA patients with the syndrome of liver depression and kidney deficiency,and the control group was 43 healthy unpregnant women.General data and stool samples were collected from two groups.Characterizing the composition of the fecal microbiota using bacterial 16S rRNA gene sequencing.The sequencing results were analyzed for diversity and species.TCM syndrome score and DASS-42 were collected from the URSA group,and the relationship between these scores and gut microbiota was analyzed.Results ①Beta diversity analysis shown that the gut microbiota of URSA patients with the syndrome of liver depression and kidney deficiency is different from that of healthy unpregnant women.②At the phylum level,Firmicutes were the most abundant in both groups.Compared with the normal group,the levels of Bacteroidetes and Proteobacteria in the URSA group were higher,and the levels of Actinobacteria were lower.At genus level,the dominant bacteria in URSA group are Faecalibacterium,Bacteroides,Megamonas,Bifidobacterium,Prevotella_9.③In the dominant bacteria of URSA group:Glucerabacter was positively correlated with age,Faecalibacterium was positively correlated with the number of abortions,Catenibacterium was negatively correlated with depression,anxiety,stress,DASS total score,Megasphaera was negatively correlated with depression,TCM syndrome score;In the dominant bacteria of normal group:Erysipelotrichaceae_UCG-003 was positively correlated with stress,the number of abortions,Bifidobacterium was negatively correlated with TCM syndrome score.Conclusion The gut microbiota composition was different between URSA patients with the syndrome of liver depression and kidney deficiency and healthy nonpregnant women,at the phylum and genus levels,the differences in gut microbiota between the two groups were observed,and the differential gut microbiota was associated with URSA patients′TCM syndrome score,suggesting that gut microbiota may be used as an indicator for the diagnosis and treatment of URSA with the syndrome of liver depression and kidney deficiency in the future,which is of clinical significance.

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.

Two Cases of Globus Sensation Successfully Treated with Ryokeijutsukanto / 日本東洋医学雑誌

Hangekobokuto is often used for globus sensation. We herein describe two cases in which ryokeijutsukanto was effective for globus sensation, for which hangekobokuto was ineffective. Case 1 was a 28-year-old woman. She had been suffering from postprandial belching, fullness and stuffy throat for a year. She underwent examination, but could not find the cause of her symptoms and visited our department. She was prescribed hangekobokuto, but the feeling of stuffiness in her throat did not improve. Case 2 was a 50-year-old woman. She had been afflicted with a feeling of fullness in the ears, tinnitus, and pain in the back of the eyes for two weeks. She visited the otolaryngology department, but no abnormalities were found so she visited our department. She was prescribed kamishoyosan, and although her ear fullness and tinnitus improved by about 60%, she began to complain of dizziness and throat stuffiness. This did not improve even though she used hangekobokuto with kamishoyosan. In both cases, after changing treatment to ryokeijutsukanto, the symptoms were improved. For globus sensation, it is necessary to select Kampo medicines while paying attention to qi deficiency, qi depression, qi counter flow, static blood, liver depression, lack of fluid, fluid toxicity, and gastrointestinal disorders. We thought that ryokeijutsukanto could be a candidate.

Establishment and evaluation of constipated irritable bowel syndrome model with liver depression syndrome / 中国实验动物学报

Objective To prepare rat models of liver stagnation syndrome constipation-type irritable bowel syndrome(IBS-C)using single and multi-factor modeling method with different indicators,to provide ideal experimental animal models of IBS-C.Methods Forty-two SD rats were divided randomly into blank(Normal),cold-water gavage(Cold),restraint(Restrain),tail-clamping(Tail),cold-water gavage + restraint(C + R),and cold-water gavage + tail-clamping groups(C + T).Body weight,food intake,water intake,and survival status,as well as open-field behavior,fecal Bristol score,visceral sensitivity,and small intestine propulsion were observed in each group during the modeling period.Pathological changes in the rat colon were observed by hematoxylin and eosin staining,and the serum and colon contents of 5-hydroxytryptamine(5-HT)and vasoactive intestinal peptide(VIP)were determined by enzyme-linked immunosorbent assay.Results The body weight in each group decreased after modeling(P<0.05,P<0.01),the food and water intakes decreased,and serum 5-HT levels increased.The number of fecal particles and Bristol score decreased while the colon 5-HT content increased in the Cold group(P<0.05,P<0.01);the total distance and average speed of the restraint group in the open field decreased(P<0.01);the preference for sugar water in the Tail group decreased(P<0.01);the preference for sugar water,total open-field distance,small intestine propulsion rate,defecation particles,and Bristol score all decreased,while the colon 5-HT content increased and the VIP content decreased in the C + T group(P<0.05,P<0.01);and the total distance,average speed,and VIP content in the colon decreased in the C + R group(P<0.05).Except for the Tail group,all the model groups showed visceral hypersensitivity(P<0.05,P<0.01)compared to the blank group at various pressure values on days 7 and 14 of modeling.Pathological observations showed no significant inflammatory cell infiltration or pathological changes in any of the model groups.Conclusions The combination of ice-water gastric lavage and tail clamping can be used to establish a rat model of liver depression syndrome in IBS-C.This may be the best of the five tested method,and the resulting model may lay the foundation for further systematic and in-depth research into the mechanism of traditional Chinese medicine in preventing and treating IBS-C.

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.

Clinical Study of Jiawei Xiaochaihutang Combined with Microwave Ablation in Treatment of Primary Hepatocellular Carcinoma / 中国实验方剂学杂志

ObjectiveTo observe the clinical efficacy of Jiawei Xiaochaihutang combined with microwave ablation （MWA） in the treatment of primary hepatocellular carcinoma （HCC） and its influence on tumor microenvironment. MethodA total of 128 patients were randomly divided into control group （64 cases： 2 cases of dropout，2 cases of elimination，and 60 cases of completion） and observation group （64 cases： 3 cases of dropout，2 cases of elimination，and 59 cases of completion）. Both groups were given comprehensive treatment after MWA surgery. Patients in control group took Biejiajian Wan orally （3 g/time，3 times/d）， and those in observation group took Jiawei Xiaochaihutang （1 dose/d）. The treatment lasted for 3 consecutive months. The size of solid tumor before and after treatment was evaluated to record the progression-free survival （PFS）. The alpha-fetoprotein-L13 （AFP-L3），des-γ-carboxy prothrombin （DCP），Golgi protein 73 （GP73），tumor necrosis factor-α （TNF-α），transforming growth factor-β （TGF-β），vascular endothelial growth factor （VEGF） and matrix metalloproteinase-2 （MMP-2） levels，as well as performance status （PS），liver function and syndrome of liver depression and Qi stagnation scores were also detected before and after treatment. In addition， the incidence of side effects of grade Ⅲ and above was compared. ResultThe total effective rate of solid tumor in observation group was 91.53% （54/59），higher than that （76.67%， 46/60） in control group（χ2=4.895，P<0.05）. The PFS in observation group was （7.16±0.95） months， longer than that （6.24±0.89 months） in control group （P<0.01）. The effective rate of traditional Chinese medicine （TCM） syndrome in observation and control groups were 88.14% （52/59）and 70.00% （42/60）， respectively （χ2=5.897，P<0.05）. The observation group （57.63%，34/59） had higher marked effective rate of TCM syndrome than control group （31.67%，19/60） （χ2=8.116，P<0.01）. The AFP-13，DCP，GP73，TNF-α，TGF-β，VEGF and MMP-2 levels and the PS，liver function and syndrome of liver depression and Qi stagnation scores in observation group were lower than those in control group （both P<0.01）. The cumulative incidence of side effects of grade Ⅲ and above in observation and control groups was 16.95% and 33.33%， respectively（χ2=4.261，P<0.05）. ConclusionConsolidation treatment of HCC after MWA surgery with Jiawei Xiaochaihutang relieved symptoms and side effects，improved PS and liver function，regulated tumor microenvironment，inhibited tumor markers and prolonged survival time. The clinical effect was better than that of Biejia decoction pill， and thus it was worthy of clinical use.

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.

Observation on efficacy of thumbtack needle combined with pediatric Tuina for constipation in children caused by liver depression and Qi stagnation / 针灸推拿医学（英文版）

Objective: To observe the clinical effect of thumbtack needles combined with pediatric Tuina (Chinese therapeutic massage) for constipation in children caused by liver depression and Qi stagnation.Methods: A total of 60 children with constipation caused by liver depression and Qi stagnation were randomly divided into a combined group and a Tuina group, with 30 cases in each group. The cases in the Tuina group were treated with pediatric Tuina, while those in the combined group were treated with additional thumbtack needle therapy. We observed the clinical efficacy and the change in the number of spontaneous bowel movements per week before and after treatment. Results: After treatment, the cure rate and the total effective rate in the combined group were higher than in the Tuina group, showing statistical significance (P<0.05). Compared with the same group before treatment, the number of spontaneous bowel movements per week in both groups increased after treatment (P<0.05). With the progression of treatment, the number of spontaneous bowel movements per week increased in both groups, and the number in the combined group was more than that in the Tuina group, showing statistical significance (P<0.01). Conclusion: Thumbtack needles combined with pediatric Tuina is more effective than pediatric Tuina alone in the treatment of constipation in children caused by liver depression and Qi stagnation.

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.

Modified Xiaoyao Erxian Decoction in Treatment of Mood Disorders in Perimenopausal Syndrome / 中国实验方剂学杂志

Objective:To observe the clinical efficacy of modified Xiaoyao Erxian decoction in the treatment of mood disorders among perimenopausal syndrome due to kidney deficiency and liver depression and its effects on monoamine neurotransmitters and brain-derived neurotrophic factor （BDNF）. Method:According to the random number table， 108 patients were divided into a control group （54 cases） and an observation group （54 cases）. Control group were treated with Shugan Jieyu capsule orally， two capsules per time， two times per day， while those in the observation group received the modified Xiaoyao Erxian decoction， one bag per day， for 12 consecutive weeks. Before and after treatment， the Hamilton Anxiety Scale （HAMA）， 17-item Hamilton Depression Rating Scale （HAMD-17）， modified Kupperman Index （KI）， Pittsburgh Sleep Quality Index （PSQI）， Menopause-specific Quality of Life （MENQOL） and kidney deficiency and liver depression syndrome scores were calculated. The levels of estradiol （E₂）， follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， BDNF， 5-hydroxytryptamine （5-HT）， norepinephrine （NE）， and dopamine （DA） were detected， followed by safety evaluation. Result:The HAMA， HAMD-17， KI， kidney deficiency and liver depression syndrome， PSQI， and MENQOL scores of the observation group were lower than those of the control group （<italic>P</italic><0.01）， whereas the 5-HT， E₂， DA， NE， and BDNF levels in the observation group were higher （<italic>P</italic><0.01）. The clinical efficacy of the observation group was superior to that in the control group （<italic>Z</italic>=2.184， <italic>P</italic><0.05）. No adverse reactions occurred after the oral administration of Chinese medicinal preparations. Conclusion:The modified Xiaoyao Erxian decoction effectively alleviates the mood disorders and other related symptoms of perimenopausal syndrome due to kidney deficiency and liver depression by regulating the monoamine neurotransmitters， BDNF， and E₂， without inducing obvious side effects.

Clinical Efficacy on Shugan Bushen Huoxue Decoction in Treating Perimenopausal Perio Syndrome (Kidney Deficiency and Liver Depression and Blood Stasis Syndrome) / 中国实验方剂学杂志

Objective:To observe the clinical efficacy and mechanism of Shugan Bushen Huoxue decoction in the treatment of perimenopausal perio syndrome （PPS） with kidney deficiency， liver depression and blood stasis syndrome. Method:One hundred and twelve patients were randomly divided into control group and observation group according to random number table. Both groups took Remifemin orally， 1 tablet/time， by swallowing in the morning and evening. The patients in control group additionally took Fuke Yangrong capsules， 4 capsules/time， 3 times/day. The patients in observation group additionally took Shugan Bushen Huoxue decoction， 1 dose/day. The course of treatment was 12 weeks in both groups. Before and after treatment， scores were graded for modified Kupperman index （KI）， traditional Chinese medicine （TCM） syndromes， menopausal quality of life scale （MENQOL）， self-rating depression scale （SDS） and self-rating anxiety scale （SAS）. Follicle stimulating hormone （FSH），luteinizing hormone （LH），serum estrogen （E₂）， 5-hydroxytryptamine （5-HT）， dopamine （DA）， norepinephrine （NE）， 5-hydroxyindole acetic acid （5-HIAA）， endothelin （ET）， and nitric oxide （NO） levels were detected before and after therapy. Result:In the observation group， scores of KI， TCM syndrome， SDS and SAS were lower than those in the control group （<italic>P</italic><0.01）. All dimensions of MENQOL scores in the observation group were lower than those in the control group （<italic>P</italic><0.01）. FSH and LH levels in the observation group were lower than those in the control group， and the E₂ level was higher than that of the control group （<italic>P</italic><0.01）. The levels of 5-HT， 5-HIAA， DA and NE in the observation group were higher than those in the control group （<italic>P</italic><0.01）. The ET level in the observation group was lower than that in the control group， and the NO level was higher than that of the control group （<italic>P</italic><0.01）. In observation group，the clinical efficacy was superior to that in control group （<italic>Z</italic>=2.073<italic>，P</italic><0.05）， and the efficacy of TCM syndromes was also superior to that in control group （<italic>Z</italic>=2.086<italic>，P</italic><0.05）. Conclusion:Shugan Bushen Huoxue decoction in the treatment of PPS in patients with kidney deficiency and liver depression and blood stasis can significantly reduce clinical symptoms， depression and anxiety， regulate the sex hormones， vasomotor factors and monoamine neurotransmitters levels， and improve the quality of life， with obvious clinical efficacy and high safety， so it is worthy of clinical use.