In the era of evidence-based medicine, it is necessary to explore the unique advantages of traditional Chinese medicine (TCM) based on standardized technical methods and operating procedures in order to achieve the modernization and internationalization of TCM and benefit all humanity. The proposal of a three-pronged evidence system combining TCM theory, human experience and experimental evidence marks an important progress in the thinking method of the TCM evaluation system. The multi-evidence body integrated through appropriate methods provides a strong support for the clinical guideline recommendations and evidence-based health decision-making in TCM. Based on the current methodological progress of international evidence synthesis and grading, this paper proposes a novel approach for integrating multi-evidence in TCM: the MERGE framework. The aim is to establish a solid foundation for the development of this methodology and provide guidance for the advancement of evidence-based medicine framework in TCM.

Objective To analyze Professor Zhang Binghou's medication experience in the treatment of diabetic kidney disease using data mining methods;To screen the core medicinal pairs and medicinal groups.Methods Prescriptions of diabetic kidney disease of Professor Zhang Binghou from the outpatient department of Beijing Hospital of Traditional Chinese Medicine Affiliated to Capital Medical University were selected from January 2014 to December 2021.TCM Inheritance Platform System 2.5,SPSS Modeler 18 and SPSS Statistics 21 software were used for association rules,clustering analysis and factor analysis to summarize the medication frequency,properties and tastes and meridians,and medicinal pairs and combinations.Results A total of 161 prescriptions were included,involving 188 kinds of Chinese materia medica,with a total frequency of 2 220 time.The kinds of Chinese materia medica with higher frequency were Rehmannize Radix et Praeparata,Testudinis Carapax et Plastrum,Astragali Radix,Rehmannine Radix,etc.The main properties were cold and warm,the main tastes were sweet and bitter,and the main meridians were kidney,liver and spleen meridians.A total of 14 drug pair association rules were obtained,with 27 commonly used drug combinations.Clustering analysis extracted 10 combinations based on the spectrum,and factor analysis extracted 14 common factors.Conclusion Professor Zhang Binghou's treatment for diabetic kidney disease takes nourishing the true yin and clearing away damp-heat as the main treatment method,and at the same time,it pays attention to tonifying kidney,consolidating essence,nourishing yin and containing yang,promoting blood circulation and removing blood stasis,etc.,which embodies Professor Zhang Binghou's unique academic thought of treating diabetic kidney disease.

ObjectiveTo investigate the effects of Yishen Daluo prescription （YSDL） on Ras homolog（Rho）/Rho-associated coiled-coil containing protein kinase（ROCK）signaling pathway in mice with experimental autoimmune encephalomyelitis （EAE） based on the silencing of β-arrestin1 gene. MethodSixty C57BL/6 female mice were randomly divided into a blank group， a model group， a virus group， a YSDL group， a virus + YSDL group， and a prednisone acetate group （hormone group）. The EAE model was induced in mice except for those in the normal group. Adeno-associated virus（AAV）solution （150 μL， 1×10¹¹ vg·mL^-1） was injected into the tail vein of each mouse in the virus group and the virus + YSDL group on the 4^th day of immunization. Drugs were administered on the 8^th day of modeling. Specifically， normal saline was given to the mice in the normal group，the model group，and the virus group at 10 mL∙kg^-1， prednisone acetate suspension to those in the hormone group at 3.9 g∙kg^-1，and YSDL to those in other groups at 20 g∙kg^-1 for 14 consecutive days. The mice were weighed and scored every day. The neurological function scores of mice in each group were recorded every day after immunization. Hematoxylin-eosin （HE） staining was used to determine the inflammatory response and lesion location in the brain tissues and spinal cord tissues of mice. The protein expression of β-arrestin1，Ras homolog gene family member A（RhoA）， and Rho-associated coiled-coil forming protein kinase Ⅰ（ROCK Ⅰ） in spinal cord and brain tissues of EAE mice was determined by Western blot. ResultCompared with the model group， the virus group and the virus + YSDL group showed decreased neurological function scores （P<0.01），and the YSDL group also showed decreased neurological function scores（P<0.05）. HE results showed that there was obvious inflammatory reaction in the central nervous system （CNS） of the model group， which was alleviated to varying degrees in other groups compared with the model group. Western blot results showed that compared with the blank group， the model group showed increased protein expression levels of β-arrestin1， RhoA， and ROCK Ⅰ in the spinal cord tissues （P<0.01）. Compared with the model group， the virus group， the YSDL group， the virus + YSDL group， and the hormone group showed decreased protein expression levels of β-arrestin1， RhoA， and ROCKⅠ in the spinal cord tissues （P<0.01）. Compared with the blank group， the model group showed increased protein expression levels of β-arrestin1， RhoA， and ROCK Ⅰ in the brain tissues （P<0.01）. Compared with the model group， the virus group， the YSDL group， the virus + YSDL group， and the hormone group showed decreased protein expression level of β-arrestin1 in the brain tissues （P<0.01）， and the virus group and the YSDL group showed decreased protein expression levels of RhoA， and ROCKⅠ in the brain tissues （P<0.05）. Additionally， the virus + YSDL group and the hormone group showed decreased protein expression levels of RhoA and ROCKⅠ in the brain tissues （P<0.01）. ConclusionYSDL can improve the clinical symptoms of EAE mice and improve the inflammatory response of CNS. The mechanism is presumably attributed to the fact that YSDL inhibits the expression of β-arrestin1 in CNS，thereby reducing the expression of Rho/ROCK signaling pathway. Furthermore， YSDL may have a synergistic effect with the inhibition of β-arrestin1 gene expression.

The automation of traditional Chinese medicine(TCM)pharmaceuticals has driven the development of process analysis from offline to online.Most of common online process analytical technologies are based on spectroscopy,making the identification and quantification of specific ingredients still a challenge.Herein,we developed a quality control(QC)system for monitoring TCM pharmaceuticals based on paper spray ionization miniature mass spectrometry(mini-MS).It enabled real-time online qualitative and quantitative detection of target ingredients in herbal extracts using mini-MS without chromatographic separation for the first time.Dynamic changes of alkaloids in Aconiti Lateralis Radix Praeparata(Fuzi)during decoction were used as examples,and the scientific principle of Fuzi compatibility was also investigated.Finally,the system was verified to work stably at the hourly level for pilot-scale extraction.This mini-MS based online analytical system is expected to be further developed for QC applications in a wider range of pharmaceutical processes.

OBJECTIVE To study the effects of Yishen daluo decoction on inflammatory factors and cyclic adenosine monophosphate（cAMP）/protein kinase A （PKA）/cAMP response element binding protein （CREB） signal pathway in experimental autoimmune encephalomyelitis （EAE） model mice by inhibiting the expressions of β-arrestin1， and to explore the mechanism of Yishen daluo decoction in the treatment of EAE. METHODS Sixty mice were randomly divided into normal group， model group， TCM group （Yishen daluo decoction 20 g/kg）， positive control group （prednisone acetate 3.9 mg/kg）， β-arrestin1 siRNA adeno- associated virus （AAV-β） group， AAV-β+TCM group， with 10 mice in each group. Except for normal group， EAE model was made in other groups. AAV-β group and AAV-β+TCM group were injected with AAV-β via tail vein to interfere with the expression of β -arrestin1 protein. Starting from the 8th day of modeling， they were given corresponding drug solution/normal saline intragastrically， once a day， for consecutive 14 days. The neurological function score of mice was detected； the pathological and morphological changes were observed in the brain and spinal cord tissues of mice； the serum levels of inflammatory factors [interleukin-2 （IL-2）， IL-23， interferon-γ （IFN-γ）] in mice were determined； the expressions of β-arrestin1， cAMP， PKA and CREB in brain and spinal cord were detected. RESULTS Compared with normal group， neurological function scores， serum levels of inflammatory factors， and protein expressions of β-arrestin1 in brain and spinal cord were significantly increased （P＜0.05 or P＜ 0.01）； protein expressions of PKA， CREB and cAMP in brain and spinal cord were decreased significantly（P＜0.05 or P＜0.01）. The deep staining of cellular shrinkage and aggregation of inflammatory cells were observed in most neurons of the brain and spinal cord， with varying degrees of demyelinating. Compared with model group， the neurological function scores， pathological changes in brain and spinal cord tissues， and most indicators （except for CREB and cAMP proteins in the brain tissue of AAV-β group） were significantly reversed （P＜0.05 or P＜0.01）.Compared with AAV- β group， the neurological function scores， the levels of IFN-γ in serum and β-arrestin1 in spinal cord were significantly decreased （P＜0.05 or P＜0.01）， PKA and cAMP in brain and spinal cord tissues were significantly increased in AAV- β +TCM group （P＜0.05 or P＜0.01）. CONCLUSIONS Yishen daluo decoction can inhibit the expression of β-arrestin1 in the central nervous system thus activating the cAMP/PKA/CREB signaling pathway， relieving nervous system inflammation， and ultimately alleviates the symptoms of EAE.

Objective:To explore the expression level of exosomal miR-218-5p in the serum of patients with colorectal cancer (CRC) and its correlation with the clinical pathological characteristics, and evaluate its diagnostic efficacy in CRC.Methods:A group of 78 patients with colorectal cancer diagnosed in Tongji Hospital affiliated to Tongji University from October 2016 to October 2018 were selected. Blood was collected before operation and serum was preserved. Forty cases of healthy people in the same period were selected as the control group. ExoQuick kit was used to extract serum exosomes. Transmission electron microscope, NTA and western blot were used to identify the morphology and molecular phenotype of exosomes. MiRNeasy kit was used to extract total RNA in serum exosomes. Real-time fluorescent quantitative PCR (RT-qPCR) was used to detect the expression level of serum exosomal miR-218-5p in each group. CRC patients were divided into high expression and low expression groups using the median relative expression level of miR-218-5p as the cut off value, and the four-square chi-square test (χ 2 test) was used to judge the relationship between miR-218-5p and clinicopathological features. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic efficacy of miR-218-5p in colorectal cancer. Results:The exosomes in serum were successfully extracted by kit method. The expression level of serum exosomal miR-218-5p in patients with colorectal cancer was significantly lower than that in normal healthy people [0.566(0.364, 0.850) vs 1.054(0.781, 1.709), P<0.001]. The low expression level was significantly better then the correlated with tumor size, TNM stage, lymph node metastasis and depth of invasion (all P<0.05). Receiver operating characteristic (ROC) analysis showed that the area under the curve (AUC) of serum exosomal miR-218-5p for the diagnosis of CRC was 0.827 (95% CI 0.754-0.900), which was significantly better than the conventional tumor marker carcinoembryonic antigen CEA (AUC =0.718, 95% CI 0.626-0.811) and carbohydrate antigen CA199 (AUC = 0.661, 95% CI 0.564-0.758). Conclusions:The down-regulation of miR-218-5p expression in serum exosomes of colorectal cancer patients is associated with a variety of adverse clinicopathological factors, which has the potential to become a diagnostic biomarker for colorectal cancer.

Objective:To investigate the effect and mechanism of lysine acetyltransferase 5(KAT5) on the radio-sensitivity of anaplastic thyroid carcinoma (ATC).Methods:The expression levels of endogenous KAT5 in ATC and normal thyroid cells were detected by Western blot and qRT-PCR. The effect of KAT5 specific inhibitor NU9056 on the radio-sensitivity of human ATC cells and normal thyroid cells was evaluated by colony formation assay. TCGA database, JASPAR database, along with Western blot, microRNA sequencing, qRT-PCR and dual-luciferase reporter assay were conducted to unravel the underlying mechanism.Results:The expression of endogenous KAT5 at the protein and mRNA levels in human ATC cells was significantly higher than that in normal thyroid cells. NU9056 could significantly enhance the radiosensitivity of human ATC cells to 8505C and CAL-62, whereas showed no sensitization effect on normal thyroid cell Nthy-ori 3-1. Knockdown of KAT5 and NU9056 both down-regulated the expression level of miR-210 in the TC cells, while NU9056 decreased the expression level of transcription factor c-Myc. The putative binding sites of c-Myc in the miR-210 promoter region were predicted, and transfection of c-Myc plasmid significantly enhanced the luciferase activity of miR-210 promoter. Elevated miR-210 level was associated with worse survival of patients with thyroid carcinoma. Down-regulated expression of miR-210 decreased the TET2 mRNA level, while inhibition of miR-210 increased the TET2 mRNA level.Conclusion:The aberrantly-activated KAT5/miR-210/TET2 pathway probably causes the radioresistance of ATC, becoming a novel sensitizing target for ATC radiotherapy in clinical practice.

OBJECTIVE: To explore the relationship between occupational aluminum exposure and fasting blood glucose level in workers. METHODS: A cluster sampling method was used to select 178 occupational aluminum-exposed workers as the exposure group, and 178 workers without occupational aluminum exposure as the control group in a large aluminum factory in Shanxi Province. Glucose oxidase method was used to measure the fasting blood glucose level, and inductively coupled plasma mass spectrometry was used to measure the plasma aluminum level in these workers. A generalized linear model was used to analyze the correlation between plasma aluminum exposure level and fasting blood glucose in these workers. RESULTS: The blood aluminum level of workers in the exposure group was higher than that of the control group [median: 39.58 vs 16.67 μg/L, P<0.01]. The fasting blood glucose level of workers in the exposure group was higher than that of the control group [(5.33±0.79) vs(5.15±0.41) mmol/L, P<0.01]. The results of the generalized linear model analysis showed that the blood aluminum level of workers was positively correlated with their fasting blood glucose level after adjusting for age, body mass index, smoking, drinking, exercise, family history of diabetes, and incidence of diabetes(P<0.05). There was a dose-response relationship between the blood aluminum level and fasting blood glucose level of workers in the groups of junior high school and below and high school(all P_(trend)<0.01). There was no correlation found between blood aluminum level and fasting blood glucose level in the group of college and above(P_(trend)>0.05). There was a dose-response relationship between the blood aluminum level and the fasting blood glucose level in the workers in the non-exercise group(P_(trend)<0.01). There was no correlation found between the blood aluminum level and the fasting blood glucose level in the exercise group(P_(trend)>0.05). CONCLUSION: The blood aluminum level of workers exposed to occupational aluminum is positively correlated with their fasting blood glucose level. Higher education level or exercise can moderately reduce the effect of blood aluminum level on fasting blood glucose.

Background and purpose:miR-196a2 functions as an oncogene during tumor initiation and pro-gression. The up-regulation promotes tumor cell proliferation, invasion and metastasis. Therefore, it is promising to be an important tumor biomarker. The aim of this study was to investigate whether rs11614913, a gene polymorphic site ofmiR-196a2, is associated with the risk of leukemia.Methods:A case-control analysis was employed. Bone marrow or periph-eral blood was collected from 210 leukemia patients diagnosed from Jan. 2009 to Jul. 2015 in Yantaishan Hospital (case group) as well as 250 healthy people who were physically examined during the same period (control group). Polymerase chain reaction-restriction fragment length polymorphism (PCR-PFLP) was used to detect the genotype of rs11614913. Application test was used to compare the difference in the frequency of each genotype between case group and control group. The odds ratio (OR) of SNP allelic genes was calculated using logistic regression analysis and 95%CI represented the risk of leukemia for each genotype.Results:The distribution differences in the frequency of T/T, C/C, C/T genotype of miR-196a2 rs11614913 between case group and control group were statistically significant (P<0.05). The risk of leukemia for individuals who carried mutant homozygous C/C was 2.661-fold higher than those carried wild-type homozygous T/T, and the difference was statistically significant (P<0.05).Conclusion:ThemiR-196a2 gene polymorphic site rs11614913 was associated with the risk of leukemia. Mutant homozygous C/C or C allelic gene carrying was probably a risk factor for leukemia.

Objective To investigate the clinical value of multidisciplinary team (MDT) assessment for chronic constipation.Methods The clinical characteristics of 346 patients with chronic constipation who were admitted to the Third Peolep's Hospital of Hangzhou from January 2010 to December 2013 were multidisciplinarily assessed.The muhidisciplinary team was involved surgeons from the department of colorectal surgery,urology,gynecology,psychiatry and psychology,and tools including questionnaires,defecography,anorectal manometry,colon transit study,urodynamic tests,gynecological examination were applied in the study.The measurement data with normal distribution were presented as (x) ± s,the comparison between groups was analyzed using the ANOVA,and the count data were analyzed using the chi-square test.Results Three hundred and forty-six patients who met criteria of this research were selected,including 86 males and 260 females with the ratio of 1 ∶ 3;the mean age was (55 ± 11)years.Of the 346 patients,slow transit constipation accounted for 7.52％ (26/346),defecatory disorder for 60.98％ (211/346),and mixed constipation for 31.50％ (109/346).A total of 93.85％ female patients (244/260) had anterior rectocele,75.43％ (261/346) patients had internal rectal mucosal prolapse,66.76％ (231/346) patients had perineum descending,23.99％ (83/346) had achalasia or inappropriate contraction of internal anal sphincter,18.79％ (65/346) had puborectalis rectocele muscle thickening,5.49％ (19/346) had rectal prolapse.A total of 82.37％ (285/346) patients were involved in other subjects than colorectal surgery.A total of 28.61％ (99/346) patients presented with urinary symptoms,including 65 cases with stress urinary incontinence,23 cases with unstable bladder and 19 cases with bladder neck obstruction (some patients had multiple urological systoms).The incidence of reproductive organ prolapse in female patients was 31.92％ (83/260),the incidence of uterine prolapse and anterior vaginal prolapse were 26.15％ (68/260) and 29.23％ (76/260),respectively.Patients with anxiety and/or depression accounted for 36.13％ (125/346).The male and female patients of slow transit constipation,defecatory constipation and mixed constipation were 10 vs 16,30 vs 79,46 vs 165,respectively,the age was 60 ± 12,56 ± 11,52 ± 10,showing no significant differences (x2=4.046,F =2.877,P ＞ 0.05).In the three kinds of constipation,patients with urinary diseases accounted for 26.92％ (7/26),26.61％ (29/109) and 29.86％ (63/211),patients with gynecological diseases accounted for 11.54％ (3/26),20.18％ (22/109),27.49％ (58/211),patients with psychological diseases accounted for 38.46％ (10/26),39.45％ (43/109),34.12％ (72/211),respectively,showing no significant difference (x2=4.090,P ＞ 0.05).Conclusion MDT assessment for patients with chronic constipation can reflect comprehensively clinical characteristics of chronic constipation,therefore multidisciplinary team should be emphasized in clinical diagnosis and treatment of chronic constipation.