[Integrated strategy for biomarkers of stable coronary heart disease with phlegm and blood stasis syndrome based on RNA-seq and network pharmacology].

This study aimed to analyze the biological foundation and biomarkers of stable coronary heart disease(CHD) with phlegm and blood stasis(PBS) syndrome based on RNA-seq and network pharmacology. Peripheral blood nucleated cells from five CHD patients with PBS syndrome, five CHD patients with non-PBS syndrome, and five healthy adults were collected for RNA-seq. The specific targets of CHD with PBS syndrome were determined by differential gene expression analysis and Venn diagram analysis. The active ingredients of Danlou Tablets were screened out from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform, and the "component-target" prediction was completed through PubChem and SwissTargetPrediction. The "drug-ingredient-target-signaling pathway" network of Danlou Tablets against CHD with PBS syndrome was optimized by Cytoscape software. After the target biomarkers were identified, 90 participants were enrolled for diagnostic tests, and 30 CHD patients with PBS syndrome were included in before-and-after experiment to determine the therapeutic effect of Danlou Tablets on those targets. As revealed by RNA-seq and Venn diagram analysis, 200 specific genes were identified for CHD with PBS syndrome. A total of 1 118 potential therapeutic targets of Danlou Tablets were predicted through network pharmacology. Through integrated analysis of the two gene sets, 13 key targets of Danlou Tablets in the treatment of CHD with PBS syndrome were screened out, including CSF1, AKR1C2, PDGFRB, ARG1, CNR2, ALOX15B, ALDH1A1, CTSL, PLA2G7, LAP3, AKR1C3, IGFBP3, and CA1. They were presumably the biomarkers of CHD with PBS syndrome. The ELISA test further showed that CSF1 was significantly up-regulated in the peripheral blood of CHD patients with PBS syndrome, and was significantly down-regulated after Danlou Tablets intervention. CSF1 may be a biomarker for CHD with PBS syndrome, and it is positively correlated with the severity of the disease. The diagnostic cut-off of CSF1 for CHD with PBS syndrome was 286 pg·mL~(-1).

Metallothionein 1M suppresses tumorigenesis in hepatocellular carcinoma.

Members of the metallothionein (MT) family are involved in metal detoxifcation and in the protection of cells against certain electrophilic carcinogens. In present study, it was found that MT1M was downregulated in more than 77.1% (91/118) of hepatocellular carcinoma (HCC) tissues compared with adjacent non-tumor tissues. Furthermore, overexpression of MT1M inhibited cell viability, colony formation, cell migration and invasion in HCC cell lines and tumor cell growth in xenograft nude mice, and activated cell apoptosis in HCC cell lines. In addition, immunohistochemistry analysis showed MT1M was negative or weak staining in tumor tissues but moderate or strong staining in adjacent non-tumor tissues. The sensitivity and specificity of MT1M for HCC diagnosis were 76.27% and 89.83%, respectively. In conclusion, MT1M was identified as a potential tumor marker for HCC and may serve as a useful therapeutic agent for HCC gene therapy.

[Study on protective and haemodynamic effects of Danshen Tongluo capsule on rats with myocardial infarction].

To study the protective effect of Danshen Tongluo capsule on rat hearts in the myocardial infarction (MI) model. After being fed with high fat diets for one month, the SD rats were randomly divided into the sham group and the model group according to the left ventricular ejection fraction (EF). The MI model group was duplicated by ligating coronary artery and then divided into five groups: the model group, the positive control group (model + captopril), the small dose group (model + Danshen Tongluo), the medium dose group (model + Danshen Tongluo) and the high dose group (model + Danshen Tongluo). Four weeks later, changes in myocardium ultra-structure were observed by hemodynamics, cardiac ultrasound and electron microscope. The results showed: (1) All doses of Danshen Tongluo capsule could significantly reduce the left ventricular end diastolic pressure (LVEDP) (P <0.01), increase the maximum internal pressure of the left ventricle (+ dp/dtmax), and lower the drop rate of left ventricular pressure (- dp/dtmax), with statistical significances in medium and high dose groups; (2) The B ultrasound results showed increase in EF and left ventricular shortening fraction (FS) in all dose groups of Danshen Tongluo capsule; (3) The medium dose group showed significant decrease in myocardial infarction index (P <0.01) and injured and fractured myofilament and sarcomere of ischemic myocardium in myocardial ultra-structure; All of Danshen Tongluo capsule-treated groups revealed reduction in myocardial injury and myocardial infraction area. The study preliminarily proves that Danshen Tongluo capsule can improve hemodynamic function, and has a protective effect on myocardial ischemia.

Effects of kaixin jieyu decoction on behavior and glial fibrillary acidic protein expression in cerebral hippocampus of a rat vascular depression model.

OBJECTIVE: To explore the effects and anti-depression mechanisms of Kaixin Jieyu Decoction (, KJD). METHODS: The rat vascular depression (VD) model was established by ligation of bilateral common carotid arteries (LBCCA) combined with chronic unpredictable mild stress (CUMS). Forty Wistar rats were randomly divided into sham, VD model, VD + high-dose KJD [15.4 g/(kg·d) of crude drug], VD + medium-dose KJD [7.7 g/(kg·d) of crude drug], and VD + fluoxetine [2.4 mg/(kg·d)] groups (n=8 in each group), and the treatments lasted for 21 days. Changes of behavior and hippocampus pathology were observed. The level of glial fibrillary acidic protein (GFAP) protein and mRNA in hippocampus was detected respectively by immunohistochemistry and real-time polymerase chain reaction. RESULTS: Compared with the sham group, rats in model group showed a variety of behavioral obstacles, including a significant reduction in sucrose consumption percentage, horizontal and vertical activity scores in open-field tests (P<0.05 or P<0.01), pathological damage like neuronal degeneration, necrosis, and a significant decrease of GFAP protein and mRNA in hippocampus (P<0.01); compared with the model group, rats in the high-dose KJD group, medium-dose KJD group and fluoxetine group obtained notable higher behavioral scores, and pathological injury lessened in hippocampus with a increased expression of GFAP protein and mRNA P<0.05 or P<0.01); compared with the medium-dose KJD group and fluoxetine group, GFAP mRNA in high-dose KJD group expressed higer (P<0.05). CONCLUSION: LBCCA combined with CUMS may cause depression-like behavioral changes resulting in the VD model of rats whose depression state can be ameliorated by KJD, and the mechanism of cerebral protection is related possibly with promoting expression of GFAP in hippocampus.

Effects of Kaixin Jieyu Decoction () on behavior, monoamine neurotransmitter levels, and serotonin receptor subtype expression in the brain of a rat depression model.

OBJECTIVE: To determine the mechanisms underlying the anti-depressant effects of Kaixin Jieyu Decoction (, KJD) by investigating the effects of KJD on behavior, monoamine neurotransmitter levels, and serotonin (5-HT) receptor subtype expression in the brain in a rat model of depression. METHODS: The rat depression model was established using chronic unpredictable mild stress (CUMS). Forty-eight Sprague Dawley rats were randomly divided into control, depression model (CUMS), CUMS+KJD (7.7 g/kg(-1)·d(-1) of crude drug), and CUMS+fluoxetine (2.4 mg/kg(-1)·d(-1)) groups (n=12 in each group), and the treatments lasted for 21 days. We regularly evaluated body weight, sucrose consumption, and horizontal and vertical activity scores in open-field tests. The content of the monoamine neurotransmitters 5-HT, norepinephrine (NE), and dopamine (DA) and the DA metabolite homovanillic acid in the cerebral cortex, and 5-HT1A and 5-HT2A receptor mRNA in the cerebral cortex and the hippocampus, were determined respectively by high-performance liquid chromatography-coularray electrochemical detector and real-time polymerase chain reaction. RESULTS: Compared with the control group, CUMS rats showed a variety of depression-like behavioral changes, including a significant reduction in body weight, sucrose consumption, and horizontal and vertical activity scores in open-field tests (P<0.05 or P<0.01), and a significant decrease in 5-HT and NE levels and 5-HT2A receptor mRNA expression. In contrast, they showed a significant increase in 5-HT1A receptor mRNA expression in the cerebral cortex. In the hippocampus, 5-HT1A receptor mRNA expression was lower whereas 5-HT2A receptor mRNA expression was higher than in the control group (P<0.05 or P<0.01). Treatment with KJD or fluoxetine partially attenuated these changes (P<0.05 or P<0.01). CONCLUSION: KJD could normalize the levels of 5-HT and NE and adjust the balance of 5-HT1A and 5-HT2A receptor expression in rat cerebrum, and this may be one of mechanisms of antidepressant effects of KJD.

[Establish proposal of diagnosis and treatment of traditional Chinese medicine in AIDS patients with recurrent oral ulcerations].

The pathogenesis of acquired immune deficiency syndrome-associated recurrent oral ulcerations (AIDS-ROU) remained obscure and these was no specific treatment for it. Syndrome differentiation in traditional Chinese medicine (TCM) focus on integral regulation and has an advantage of the disease that etiology and pathogenesis remain obscure. A draft of Chinese medicine diagnosis and treatment standard procedure for AIDS-ROU was established by literature retrieval and peer review. Two questionnaires were carried out to investigate the confirmation and advice of in-group specialist to key points of the draft including diagnosis, treatment and nursing. Then the procedures were revised accordingly. The preliminary results showed the recovery rate of complete questionnaires in the 1st survey was 96%. Specialists confirmed more on outline, case history and physical examinations, syndrome differentiation of hyperactivity of fire due to Yin deficiency syndrome (HFYDS), treatment of heat accumulated in heart and spleen syndrome (HAHSS) and HFYDS, treatment of western medicine and nursing. They held different opinions on incidence, treatment of deficiency of spleen-QI and stomach-QI syndrome (DSSS) and criterion of therapeutical evaluation. Cronbach coefficient alpha (CCA) was 0.998 and split-half reliability R was 0.91. Recovery rate of complete questionnaires in 2nd survey was 100%. Specialists confirmed more on outline, etiology and pathogenesis, case history and physical examination, auxiliary examination, diagnostic criteria, syndrome differentiation and treatment of HAHSS and HFYDS. They held different opinions on syndrome differentiation and treatment of intermingled cold and heat syndrome and DSSS, nursing and the other therapies. CCA was 0.428 and split-half reliability R was 0.96. Coefficient of variations of the 2nd survey were less than those of the 1st survey, which mean coordination was improved. Each single item in two surveys contributed less difference in overall results according to weight coefficients. A new revision of the procedure was preliminarily established according to results of two surveys. Experts'activeness, concentration and coordination were good in two surveys. They had reached consensus in key points of the draft including diagnosis, treatment and nursing on the whole.

[Construction and analysis of questionnaires on AIDS cough in traditional Chinese medicine diagnosis and treatment procedures].

OBJECTIVE: To norm the behavior of AIDS cough in traditional Chinese medicine diagnosis and treatment and improve the clinical level of cough treatment for HIV/AIDS, and build AIDS cough diagnosis and treatment procedures in traditional Chinese medicine. METHOD: Combined with clinical practice,to formulate questionnaire on AIDS cough in traditional Chinese medicine diagnosis and treatment by both English and Chinese literature research to expertise consultation and verify the results of the questionnaires on the statistics using the Delphi method. RESULT: Questionnaire contents consist of overview, pathogeny, diagnosis standard, dialectical medication (phlegm heat resistance pulmonary lung and kidney Yin deficiency lung spleen-deficiency), treating spleen-deficiency (lung), moxibustion treatment and aftercare care and diet and mental, average (2.93-3.00), full mark rate (93.10%-100%) ranks average (9.91-10.67) and (287.50-309.50) of which are the most high value, and the variation coefficient is 0.00, the Kendall coefficient (Kendalls W) is 0.049 which is statistical significance, the questionnaire reliability value of alpha was 0.788. CONCLUSION: Preliminary standarded concept, etiology and pathogenesis, diagnosis and syndrome differentiation treatment of AIDS cough, basically recognised by the experts in this field, and laid the foundation of traditional Chinese medicine diagnosis and treatment on develop the AIDS cough specifications.

[A research of Chinese medicine diagnosis and treatment standard procedure for patients with HIV associated pruritus based on experts' questionnaires].

OBJECTIVE: To establish Chinese medicine diagnosis and treatment standard procedure for patients with HIV associated pruritus. METHODS: A Chinese medicine diagnosis and treatment standard procedure for patients with HIV associated pruritus was established by literature retrieval and peer review. Two questionnaires were carried out to investigate the confirmation and advice of in-group specialists to key points of the draft including diagnosis, treatment and nursing. Then the procedures were revised accordingly. RESULTS: The recovery rate of complete questionnaires in the 1st survey was 96%. Specialists confirmed more on case history and physical examinations, syndrome differentiation of three syndrome types, treatment of blood deficiency wind dryness syndrome (BDWDS) and nursing. They held different opinions on the outlines, auxiliary examinations, treatment of blood heat induced wind evil syndrome (BHWES) and wind cold and dampness accumulation syndrome (WCDAS), of which the coefficient of variations (CVs) was within 0.1603 -0.2473. The procedures were revised and the 2nd survey was launched. The recovery rate of complete questionnaires in the 2nd survey was 100%. Specialists confirmed more on case history and physical examinations, diagnostic criteria, syndrome differentiation of BDWDS and WCDAS, and treatment of BDWDS, of which CVs was 0. All indicated high agreement and good compliance. The CVs of other items were within 0.0638-0.1439, less than those of the 1st survey. The consistency of experts' opinions were somewhat improved. The contribution by one single item showed less difference in assessing the overall results in the two surveys. A new revision of the procedure was preliminarily established according to results of two surveys. CONCLUSIONS: Experts' activeness, concentration, and coordination were good in the two surveys. They had reached consensus in key points of the draft including diagnosis, treatment, and nursing.

[Diagnosis and treatment standard operating procedure of traditional Chinese medicine for HIV/AIDS diarrhea by questionnaire building].

OBJECTIVE: To establish the diagnosis and treatment standard operating procedure of Chinese medicine for HIV/AIDS diarrhea and to carry out preliminary assessment. METHODS: Under the guidance of Chinese medical theory, domestic and foreign literatures correlated to the diagnosis and treatment of HIV/AIDS diarrhea by Chinese medicine were summarized. The specialist questionnaire of clinical diagnosis and treatment standard operating procedures of Chinese medicine for HIV/AIDS diarrhea was designed by focus group discussions, and the national specialist questionnaire survey was carried out twice. RESULTS: The standard operating procedure of clinical diagnosis, treatment, nursing,and therapeutic efficacy assessment of Chinese medicine for HIV/AIDS diarrhea was preliminarily established. CONCLUSIONS: In the regulations, the concepts of the disease and symptoms and their pathogenesis were defined, and the standardization of Chinese medicine diagnosis and treatment was highlighted. Meanwhile, emphasized were the features of propaganda and education, follow-ups, consecutive diagnosis and treatment outside or inside hospitals.

[Experiment of effective components from traditional Chinese herbs on ischemic left ventricular remodeling].

OBJECTIVE: To investigate the effects of effective components extracted from medicines compatibility on ischemic myocardial remodeling. METHOD: The animal models of ischemic myocardial remodeling was copied by ligating the left anterior descending branch of coronary artery in rat. And then decoction and effective components extracted from the same medicines were administrated to rats. The heart function was detected by ultrasound diagnostic apparatus and the heart morphology was examined by Hematoxylin and Eosin, picrosirius staining respectively on the 3rd and 56rd day. RESULT: Both concentrated decoction and effective components compatibility showed good restrained effects on ventricle malignant remodeling, such as improved heart function, reduced the area of myocardial infarction, lowered heart index, mitigated the fibrosis level and decreased apoptosis cell numbers in infarction and non-infarction areas. The difference were statistically significant when compared with the control group (P < 0.01, P < 0.05), and effective components treated group had better effects than concentrated decoction group (P < 0.05). Concentrated decoction group showed little effects on the 3rd day, while effective components group showed significant effect (P < 0.05). CONCLUSION: Effective components compatibility has better effects on inhibiting the process of ischemic myocardial remodeling than concentrated decoction. Effective components compatibility has better effects on ventricle malignant remodeling by eliminating the interfrence non-pharmacodynamic elements of the medicine.