ObjectiveTo explore the possible mechanism of Yigan Fupi Prescription （抑肝扶脾方， YFP） in treating diarrhea-type irritable bowel syndrome （IBS-D） by investigating the AKT/mTOR signaling pathway. MethodsSixty SD rats were randomly divided into control group， model group， YFP low-， medium-， and high-dose group， and pinaverium bromide group， with 10 rats in each group. All groups but the control group， were subjected to 21 days of tail-clamping stimulation and 14 days of senna leaf gavage to establish a liver stagnation and spleen deficiency-type IBS-D rat model. After successful modeling， the YFP low-， medium-， and high-dose group were administered 0.96， 1.93， and 3.87 g/（kg·d） of the prescription， respectively. The pinaverium bromide group was given 13.5 mg/（kg·d）， while the control and model groups were given 10 ml/（kg·d） distilled water. All groups were administered once daily for 14 consecutive days. General conditions of the rats were recorded during the experiment， and after modeling and drug administration， body weight， Bristol stool score， abdominal withdrawal reflex （AWR） score， and histo pathology of colon tissue were observed under HE staining. ELISA was used to detect serum levels of tumor necrosis factor α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6）. Immunofluorescence was employed to detect the levels of AKT/mTOR pathway-related proteins including phosphorylated AKT （p-AKT）/AKT and phosphorylated mTOR （p-mTOR）/mTOR in the colon tissue. Western Blotting was used to detect the levels of autophagy-related proteins， including UNC-51-like kinase 1 （ULK1）， Beclin1 and LC3， and tight junction proteins including Occludin and ZO-1 in the colon tissue. ResultsAfter modeling， compared to the control group， the body weight of rats in the other groups decreased， and Bristol stool scores， as well as AWR scores under 20， 40， 60， and 80 mmHg increased （P＜0.05 or P＜0.01）. After drug administration， compared to the control group， the model group showed reduced body weight， decreased ULK1， Beclin1， LC3Ⅱ/LC3Ⅰ， Occludin， and ZO-1 protein levels in the colon tissue （P＜0.05 or P＜0.01）， and increased Bristol stool scores， AWR scores， serum TNF-α， IL-1β， and IL-6 levels， as well as p-AKT/AKT and p-mTOR/mTOR protein relative expression levels （P＜0.05 or P＜0.01）. Pathological results showed a significant reduction in goblet cells in the upper part of the glandular layer of the colon， with mild inflammatory cell infiltration. The submucosal collagen fibers were dissolved， with unclear boundaries， pale staining， and microvascular congestion and dilation. Compared with the model group， the YFP low-， medium-， and high-dose group and the pinaverium bromide group showed increased body weight， Beclin1， Occludin， and LC3Ⅱ/LC3Ⅰ protein levels （P＜0.05 or P＜0.01）， and decreased Bristol stool scores， AWR scores under 40， 60， and 80 mmHg， serum IL-1β， IL-6， TNF-α levels， and p-AKT/AKT， p-mTOR/mTOR protein relative expression levels （P＜0.05 or P＜0.01）. The pathological morphology of the rats in the YFP groups and pinaverium bromide group showed varying degrees of improvement. Compared with the pinaverium bromide group， the YFP low- and medium-dose group showed increased AWR scores under 20， 40， and 60 mmHg （P＜0.05）. The YFP low-dose group had reduced TNF-α， IL-1β， and IL-6 levels， and increased p-mTOR/mTOR protein relative expression levels occured in all YFP groups （P＜0.05）. Compared with the YFP low-dose group， the YFP high-dose group and pinaverium bromide group showed decreased AWR scores under different pressure levels and reduced p-AKT/AKT protein relative expression levels， while the YFP medium- and high-dose group had elevated serum TNF-α， IL-1β levels and reduced p-mTOR/mTOR protein relative expression levels （P＜0.05）. ConclusionYFP can effectively improve the pathological injury of colon tissue in IBS-D model rats with liver stagnation and spleen deficiency， reduce Bristol stool and AWR scores， and its mechanism may be related to reducing level of inflammatory factors and inhibiting AKT/mTOR pathway-related proteins in colon tissue， thereby enhancing the expression of autophagy-related proteins in the colon tissue.

BACKGROUND: Epoxyeicosatrienoic acids (EETs), which are metabolites of arachidonic acid catalyzed by cytochrome P450 epoxygenase, are degraded into inactive dihydroxyeicosatrienoic acids by soluble epoxide hydrolase (sEH). Many studies have revealed that sEH gene deletion exerts protective effects against diabetes. Vascular calcification is a common complication of diabetes, but the potential effects of sEH on diabetic vascular calcification are still unknown. METHODS: The level of aortic calcification in wild-type and Ephx2-/- C57BL/6 diabetic mice induced with streptozotocin was evaluated by measuring the aortic calcium content through alizarin red staining, immunohistochemistry staining, and immunofluorescence staining. Mouse vascular smooth muscle cell lines (MOVAS cells) treated with β-glycerol phosphate (0.01 mol/L) plus advanced glycation end products (50 mg/L) were used to investigate the effects of sEH inhibitors or sEH knockdown and EETs on the calcification of vascular smooth muscle cells, which was detected by Western blotting, alizarin red staining, and Von Kossa staining. RESULTS: sEH gene deletion significantly inhibited diabetic vascular calcification by increasing levels of EETs in the aortas of mice. EETs (especially 11,12-EET and 14,15-EET) efficiently prevented the osteogenic transdifferentiation of MOVAS cells by decreasing nidogen-2 (NID2) expression. Interestingly, suppressing sEH activity by small interfering ribonucleic acid or specific inhibitors did not block osteogenic transdifferentiation of MOVAS cells induced by β-glycerol phosphate and advanced glycation end products. NID2 overexpression significantly abolished the inhibitory effect of sEH gene deletion on diabetic vascular calcification. Moreover, NID2 overexpression mediated by adeno-associated virus 9 vectors markedly increased insulin-like growth factor 2 (IGF2) and phospho-ERK1/2 expression in MOVAS cells. Overall, sEH gene knockout inhibited diabetic vascular calcification by decreasing aortic NID2 expression and, then, inactivating the downstream IGF2-ERK1/2 signaling pathway. CONCLUSIONS sEH gene deletion markedly inhibited diabetic vascular calcification through repressed osteogenic transdifferentiation of vascular smooth muscle cells mediated by increased aortic EET levels, which was associated with decreased NID2 expression and inactivation of the downstream IGF2-ERK1/2 signaling pathway.
Animals ; Mice ; Vascular Calcification/metabolism* ; Mice, Inbred C57BL ; Epoxide Hydrolases/metabolism* ; Diabetes Mellitus, Experimental/genetics* ; Male ; Gene Deletion ; MAP Kinase Signaling System/genetics* ; Cell Line ; Immunohistochemistry ; Muscle, Smooth, Vascular/metabolism* ; Signal Transduction/genetics* ; Mice, Knockout

Objective: Progressive supranuclear palsy (PSP) involves a variety of visual symptoms that are thought to be partially caused by structural abnormalities of the retina. However, the relationship between retinal structural changes, disease severity, and intracranial alterations remains unknown. We investigated distinct retinal thinning patterns and their relationship with clinical severity and intracranial alterations in a PSP cohort. Methods: We enrolled 19 patients with PSP (38 eyes) and 20 age-matched healthy controls (40 eyes). All of the participants underwent peripapillary and macular optical coherence tomography. Brain 11C-2β-carbomethoxy-3β-(4-fluorophenyl) tropane (11C-CFT) and 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography imaging were also performed in patients with PSP. We investigated the association between retinal thickness changes and clinical features, striatal dopamine transporter availability, and cerebral glucose metabolism. Results: The peripapillary retinal nerve fiber layer (pRNFL) and macula were significantly thinner in patients with PSP than in controls. The thickness of the superior sector of the pRNFL demonstrated a significant negative relationship with the Movement Disorder Society-Unified Parkinson’s Disease Rating Scale part III and Hoehn and Yahr staging scale scores. A significant negative correlation was found between outer inferior macular thickness and disease duration. Outer temporal macular thickness was positively correlated with Montreal Cognitive Assessment scores. In PSP, lower outer temporal macular thickness was also positively correlated with decreased dopamine transporter binding in the caudate. Conclusion The pRNFL and macular thinning may be candidate markers for monitoring disease severity. Additionally, macular thinning may be an in vivo indicator of nigrostriatal dopaminergic cell degeneration in PSP patients.

Objective: To examine the eating behaviors and associated factors among preschool children from multiple child families, so as to provide a basis for promoting healthy eating habits in this population. Methods: From April to June 2022, a stratified random cluster sampling method was employed to selected 2 647 guardians of preschool children in 11 kindergartens in Tongling City. Parenting style questionnaire, Identification and Management of Feeding Difficulty Questionnaire (IMFeD), Caregiver s Feeding Style Questionnaire (CFSQ) and a self designed questionnaire were administered. Multiple linear regression was performed to analyze the factors influencing eating behaviors among preschool children. Results: In the study, the detection of eating behavior problems among preschool children was 71.4%, and the detection of reduced appetite was the highest (55.6%). The detection rates of anorexia, picky eating, poor eating and improper eating behavior in only child families were higher (60.6%, 41.0%, 32.0%, 19.5%), compared with those in multiple child families (51.7%, 36.4%, 27.2%, 16.6%) ( χ 2=20.05, 5.95, 7.16, 3.92, P < 0.05 ). Among multiple child families, the detection rates of fear of eating and underlying disease were the lowest in 5-<10-year-old group (2.3%, 0.6%) ( χ 2=11.54, 8.51, P <0.05). In multiple child families, the detection rates of picky eating and poor eating habits for the first-born child were higher (42.0%, 32.5%), compared with second born and third born child (34.6%, 25.6%)( χ 2= 6.15, 6.38, P <0.05). The results of multiple linear regression analysis showed that multiplechild families ( β =-1.40), democratic parenting style ( β =-0.07), higher feeding response scores ( β =-0.33) were negatively associated with eating behavior problems among preschool children ( P <0.05). Eating behavior problems among preschool children were positively correlated with less educated mothers [high and junior high school education ( β =0.87), primary school education and below ( β =3.69)], longer average daily screen time ( β =0.10), higher feeding requirements scores ( β =0.64), doting parenting styles ( β =0.21), permissive parenting styles ( β =0.27) and inconsistent parenting styles ( β =0.14)( P <0.05). Conclusion The detection rate of the eating behaviors problems among preschool children is high in schools. Greater attention should be paid to only child and first born child of multiple child families, and appropriate feeding and parenting styles, so as to promote healthy eating habits among this population.

Objective: To explore the emotional and behavioral problems of different only child and multiple child families, and to provide a reference for developing effective parenting styles. Methods: A stratified random cluster sampling method was adopted to investigate 2 647 guardians of preschool children in Tongling City from April to June, 2022. Children s emotional and behavioral problems were evaluated by using the self designed parenting questionnaire and the Children s Strengths and Difficulties Questionnaire (SDQ) parent edition. Results: The abnormal total score detection rate of emotional and behavioral problems of preschool children was 15.5%, and the rate of abnormal peer interaction was the highest (19.5%). In multiple child families, the first born child (17.5%, 20.4%), compared with the second and third child (11.5%, 9.5%), was more likely to exhibit conduct and hyperactivity problems ( χ 2=8.44, 29.75, P <0.01). There were differences in parenting attitudes between only child and multiple child families( χ 2=9.38, P <0.05). The results of the Logistic regression analysis showed that more frequent parent child discussion, the persuasive education and consistent family discipline strategy were negatively related to the emotional and behavioral problems of only children and children in multiple child families ( OR =0.15, 0.49, 0.38; 0.34, 0.40, 0.42, P <0.05). However, harmonious family relationships were only negatively related to emotional and behavioral problems in only children ( OR =0.08, P <0.01), and a higher education level among mothers was negatively related to emotional and behavioral problems in children from multiple child family environments( OR=0.30, 0.45, P <0.05). Conclusion The emotional and behavioral problems are serious of preschool children in Tongling City, the psychological development of the oldest children from multiple child and only child families should be actively followed, as this would help to promote a better understanding of the development of preschool children s physical and mental health.

Objective:To investigate the efficacy of venetoclax-based combined regimen in treatment of adult patients with acute myeloid leukemia (AML).Methods:The data of 50 adult AML (non-acute promyelocytic leukemia) who received venetoclax-based combined regimen in the First Affiliated Hospital of Xiamen University, Dongguan People's Hospital, the First Hospital of Longyan City, Jieyang People's Hospital from December 2018 to May 2021 were retrospectively analyzed. Different doses venetoclax combined with demethylation drugs or low-dose chemotherapy regimen were used to analyze the therapeutic efficacy. The related factors influencing efficacy were analyzed by using logistic regression.Results:The composite complete remission (CR) rate of 50 AML patients was 62.0% (31/50), the overall response rate (ORR) was 76.0% (38/50); 28 patients achieved effectiveness [CR and partial remission (PR)] after the first cycle and could achieve effectiveness by 3 courses of treatment at the latest. Among 50 patients, 28 cases were newly diagnosed AML, the composite CR rate was 60.8% (17/28), ORR was 78.6% (22/28); 22 cases were recurrent and relapsed, the composite CR rate was 63.6% (14/22), ORR was 72.7% (16/22); and there was no statistically significant difference of ORR between the both groups ( χ2 = 0.23, P = 0.743). Logistic regression multivariate analysis showed age was the only independent influencing factor for the treatment effectiveness ( OR = 8.451, 95% CI 1.306-54.697, P = 0.025). The median duration time of patients receiving venetoclax treatment regimen was 4.5 months (1.1-15.0 months); 16 cases who had treatment effectiveness finally relapsed, the median time of maintaining effectiveness was 5 months (1.1-11.0 months). Additionally, the common treatment-related adverse reactions included bone marrow suppression after treatment, followed by some gastrointestinal reactions like nausea, vomiting and stomachache. In addition, no patient stopped medication for more than 1 week due to bone marrow suppression related complications. Conclusion:Venetoclax-based combined regimen shows a good short-term efficacy in treatment of AML. It is also effective and tolerable for elderly patients receiving reduced dose therapy.

Objective:To investigate the risk factors for recurrence of Graves′ disease after withdrawal of antithyroid drugs (ATD).Methods:This prospective study recruited 285 patients with newly onset Graves′ disease taking ATD from 2012 to 2018 at Department of Endocrinology, Affiliated Hospital of Integrated Traditional Chinese and Western Medicine. A total of 121 patients who completed follow-up were enrolled and were divided into relapse and remission group according to whether hyperthyroidism reoccurred within 2 years after ATD with drawal. Demographics, clinical manifestations, thyroid serological characteristics, and thyroid color doppler ultrasound at baseline and withdrawal were compared between the two groups. Cox regression analysis was used to analyze the correlation between above factors and recurrence of Graves′ disease.Results:Sixty-five patients relapsed within 2 years after drug withdrawal. Patients with early recurrence were characterized by Graves′ disease genetic history and high baseline thyrotrophin receptor antibody (TRAb) levels. Family history, higher serum FT 3(≥18.1 pmol/L), FT 4(≥49.8 pmol/L), and TRAb(≥16.1 mIU/mL) levels, larger goiter(Ⅱ-Ⅲ) and thyroid volume(≥28.6 cm 3), higher peak velocity of superior thyroid artery (STA-PV; ≥0.6 m/s) before treatment, and higher TRAb(≥0.8 mIU/mL) level after ATD withdraw were risk factors for Graves′ disease recurrence. Higher 25-hydroxy vitamin D(≥14.7 ng/mL) level at baseline, as well as high level of TSH(1.4 μIU/mL) at withdrawal may reduce the risk of relapse. Conclusions:Family history of Graves′ disease, clinical manifestations, thyroid serological indicators and imaging characteristics of severe Graves′ disease before treatment all increased the risk of Graves′ disease recurrence. Patients with aforementioned factors should be actively evaluated in order to choose treatment modalities reasonably. We recommended to maintain lower TRAb titer within normal reference range and TSH level between 1.4 μIU/mL and upper limits of normal reference range at ATD withdrawal to reduce the recurrence rate of Graves′ disease.

Objective The effect of serum exosome isolation method on obtained exosomal RNA is not yet clear .The aim of this study was to provide evidences to selected exosome isolation methods .Methods This was a method comparison study to assess three methods .Vein blood samples were collected from 3 healthy donors from Nanfang hospital , 4 ℃ 12 h was taken to make blood natural coagulated and the supernatant was taken as the serum . Exosomes extracted by Ultracentrifugation , ExoQuick and Total Exosome Isolation Reagent ( TEI ) kits from serum were assessed by transmission electron microscopy , nanoparticle tracking analysis and protein analysis to identify morphology , protein expression , concentration and size distribution of particles .ExoRNA extracted by Trizol-LS, SeraMir, Total Exosome RNA Isolation ( TER) and exoRNeasy were assessed by Bioanalyzer 2100 and qPCR to ascertain RNA distribution and miRNA expression.Results For exosomes isolation , two commercial kits ( ExoQuick and TEI ) showed higher exosomes recovery and protein concentration than ultracentrifugation , but exosome isolated by ultracentrifugation expressed abundant protein makers mostly .For exo-RNA isolation, the distribution of RNA and expression of miRNAinfluenced by three methods , but there was no effect on the relative expression trend of miRNA.ExoRNeasy resulted in the highest yield and most narrow size distribution pattern of small RNA with higher level of miRNA expression .Results of TEI with TER kits showed no obvious bands of small RNA and moderate miRNA expression among methods .Ultra with Trizol-LS or ExoQuick with SeraMir showed low concentration measurable bands around 100 nt, with changeable miRNA profiling irregularly . Conclusions Extraction of exosomes using traditional ultracentrifugation method is applicable for proteomics research work .Extraction using ExoQuick and TEI kits is suitable for preparing exosomes from scarce samples such as serum.

The modern statistical and data mining technology is used to analyze the law of the acupoint selection in the treatment of insomnia with acupuncture and moxibustion. The clinical literature on the treatment of insomnia with acupuncture and moxibustion in recent 10 years is collected from China National Knowledge Infrastructure (CNKI). Excel software is applied to establish the database of acupuncture prescriptions for insomnia so as to conduct the descriptive analysis, association rule analysis and clustering analysis on the data. In the treatment of insomnia with acupuncture and moxibustion, Shenmen (HT 7) is of the highest frequency and most acupoints were selected from the Governor Vessel. The commonly used acupoints are located on the head, face, neck and nape region. The combination of the local acupoints with the distal ones is predominated. The crossing points among the specific points present the advantage in the treatment. The association: rule analysis indicates the highest, correlation among Taixi (KI 3), Sishencong (EX-HN 1) and Shenmen (HT 7). The clustering analysis results in 6 effective clusters and 10 pairs of key points and summarizes the common law of the acupoint combination for insomnia. All of these provide the reference to the treatment of insomnia with acupuncture and moxibustion.
Acupuncture Points ; Acupuncture Therapy ; China ; Data Mining ; Databases, Factual ; Humans ; Moxibustion ; Sleep Initiation and Maintenance Disorders ; therapy ; Software

This study was aimed to analyze acupoint selection principle for acute gouty arthritis treatment. Literatures on acupuncture treatment of acute gouty arthritis were collected from CNKI in recent 20 years. Excel software was used in the establishment of database on main acupoints for acute gouty arthritis treatment. On Windows 8 platform, SPSS 17.0 and Clementine 12.0 software were used in the descriptive analysis, association rules analysis and cluster analysis. The results showed that the top three acupionts with the highest frequency of application were Sanyinjiao (SP 6), Zusanli (ST 36), and Y inlingquan (SP 9). The meridian used with the highest frequency was the spleen meridian of foot Taiyin. The result of association rules analysis showed that Sanyinjiao (SP 6)-Xuehai (SP 10)-Quchi (LI 11) was the most relational one. The cluster analysis indicated the top four core cluster groups including Xiangu (ST 43)-Neiting (ST 44)-Xingjian (LR 2), Xuehai (SP 10)-Hegu (LI 4)-Quchi (LI 11), Taibai (SP 3)-Y inbai (SP 1)-Dadu (SP 2)-Fenglong (ST 40), and Zhubin (KI 9)-Zhigou (TE 6)-Gongsun (SP 4)-Bafeng (EX-LE 10). It was con-cluded that the clinical acupoint selection and combination principle in acute gouty arthritis treatment were analyzed through large amount of data. It can provide evidences and references for acupuncture clinical practice.