Objective To explore the role of neogambogic acid in the characteristics of colorectal cancer stem cells (CRC-CSCs) through the Wnt/β-catenin signaling pathway. Methods The colorectal cells SW480 and HCT166 were divided into control group and neogambogic acid groups (1.5, 3, 6, and 12 μmol/L). The viability of CRC-CSCs was determined by MTT method, and spheroid and clone formation assays were used to assess the capacity of spheroid formation and self-renewal ability of the cells. The effects of neogambogic acid on the apoptosis and cell cycle of CRC-CSCs were evaluated by flow cytometry assays. Real-time quantitative PCR was used to detect the mRNA expression levels of relative markers (CD133, CD44, ALDH1, Oct4, and Nanog) of CRC-CSCs, and the protein expression levels of the self-renewal marker (PCNA), apoptosis markers (cleaved caspase-3 and cleaved caspase-9), and Wnt/β-catenin signaling pathway markers (p-GSK3β, GSK3β, β-catenin, and Wnt) were analyzed using Western blot. Results Compared with the control group, after neogambogic acid treatment, the viability of SW480 and HCT116 cells decreased (P<0.05), the spheroid forming ability and the clone numbers of CRC-CSCs decreased (P<0.001, P<0.01) but the cell apoptosis rate increased (P<0.01), and cell cycle was arrested in G₀/G₁ phase. Moreover, neogambogic acid downregulated the mRNA and protein expression of relative markers of CRC-CSCs (CD133, CD44, ALDH1, Oct4, and Nanog), PCNA, p-GSK3β, β-catenin, and Wnt (P<0.05) and upregulated the expression of cleaved caspase-3, cleaved caspase-9, and GSK3β (P<0.01). Conclusion Neogambogic can inhibit the stem cell properties of colorectal cells via inhibition of Wnt/β-catenin signaling pathway. As a result, neogambogic acid may be an attractive agent against colorectal cancer.

Objective To analyze and discuss the medication rule of professor Ruan Yan in the treatment of children with allergic rhinitis by using data mining method,and to provide reference for the clinical research and patented drugs development for the treatment of children with allergic rhinitis.Methods The outpatient medical records of professor Ruan Yan for the treatment of children with allergic rhinitis were collected.Microsoft Excel 2010 software was used for frequency statistics.SPSS Clementine 12.0 software was used for association rule analysis,cluster analysis and factor analysis to obtain the data.The frequency of use of various drugs and the association rules between drugs were obtained.Then the medication rules in professor Ruan Yan's prescription were analyzed.Results A total of 308 Chinese medicine compounds were included,involving 80 kinds of Chinese medicines,among which relieving drugs and qi-invigorating herbs were high-frequently used.The distribution of traditional Chinese medicine syndrome types was mainly characterized by lung-qi deficiency-cold syndrome and lung-spleen qi deficiency syndrome.The medicinal properties were mainly spicy,warm and sweet,and most of them belonged to the lung,spleen and stomach meridians.Five core prescriptions were extracted by factor analysis.Four drug combinations were obtained by systematic cluster analysis.Conclusion Ventilating lung and opening the orifices,expelling wind and removing cold,strengthening the spleen and replenishing qi are basic therapeutic principles for professor Ruan Yan in the treatment of children with allergic rhinitis.The treatment mainly focused on dispelling evil,ventilating lung and opening the orifices,expelling wind and removing cold during the acute stage of allergic rhinitis.In the remission period,according to the principle of"treating disease must be based on its origin",the treatment should enhance children's physical fitness,tonify lung and strengthen spleen,thereby reducing recurrence.

Methods:Cultured human alveolar epithelial A549 cells were assigned into LPS group and blank control group. LPS group was stimulated with LPS and adenosine triphosphate to induce pyroptosis and inflammation. A549 cells were divided into 4 groups: miR-20a mimics group, mimics-negative control (NC) group, inhibitor group and inhibitor-NC group. MiRNA-20a mimics, mimics-NC, inhibitor, and inhibitor-NC were transfected respectively into A549 cells, and after 24 h, the cells were collected to verify transfection efficiency by qPCR. MiRNA-20a mimics and the constructed TLR4-3'UTR double luciferase reporter plasmid were co-transfected into A549 cells, and luciferase activity was analyzed. MiRNA-20a mimics/inhibitors were transfected into A549 cells, and then the cells were stimulated by LPS for 8 h followed by adenosine triphosphate for 30 min. QPCR, Western Blot and ELISA were used to detect the expression of GSDMD, inflammatory factors (ASC, NLRP3, Caspase-1, IL-1β) and Signaling molecules (TLR4、NF-κB) in A549 cells at mRNA level and protein level. Immunofluorescence was used to detect the expression of TLR4 in the A549 cells and NF-κB in the nucleus of A549 cells after transfecting with miRNA-20a mimics/inhibitor.Results:The mRNA and protein expression of pyroptosis marker molecule (GSDMD) and inflammatory factors (ASC, NLRP3, Caspase-1, IL-1β) in A549 cells stimulated with LPS were significantly higher than those in the blank control group, and the differences were statistically significant ( P<0.05). The expression of miRNA-20 in the mimics group was significantly higher than that in the mimic-NC group ( P<0.05), while the expression of miRNA-20a in the inhibitor group was lower than that in the inhibitor-NC group ( P<0.01). The double luciferase reporter gene experiment showed that the relative fluorescence value of the co-transfection group for TLR4-3'UTR-WT and miRNA-20a mimics was significantly lower than the co-transfection group for TLR4-3'UTR-WT and miRNA-20a mimics-NC ( P<0.05). The mRNA and protein levels of pyroptosis marker molecule (GSDMD) , inflammatory factors (ASC, NLRP3, Caspase-1, IL-1β) and signaling molecules (TLR4, NF-κB) were decreased in the mimics group compared to the mimics-NC group, and increased in inhibitor group compared to inhibitor-NC group. Conclusions:miRNA-20a may inhibit LPS-induced pyroptosis and inflammation of A549 cells via TLR4/NF-κB signal pathway.Objetive:To explore the potential role of miRNA-20a in lipopolysaccharide (LPS) induced pyroptosis and inflamation of human alveolar epithelial A549 cells and its regulation mechanisim.

Objective To explore the effect of intermittent fasting on blood glucose control in patients with type 2 diabetes mellitus(T2DM)complicated by overweight and obesity.Methods 46 T2DM patients with overweight and obesity were divided into control group(Con,n=22)and intermittent fasting group(IF,n=24)according to their wishes.Con group was given routine diet and drug treatment by specialists in endocrinology.IF group completed the intermittent fasting by using a combination of meal replacement and natural diet,and hypoglycemic drugs were reduced on fasting days.Results Compared with Con group,IF group showed a significant decrease in body weight,BMI,hypoglycemic drug efficacy score,WC,fat index,hip circumference and visceral fat(VF)(P＜0.05).After intervention,the proportion of diabetes in remission or near remission in IF group was higher than that in Con group(P＜0.05).Conclusion Intermittent fasting can reduce the application of hypoglycemic drugs in T2DM patients with overweight and obesity,and can also reduce weight and VF.

Objective:To search for and summarize the best evidence on non-pharmacological interventions for orthostatic hypotension in patients with Parkinson disease.Methods:Based on the 6S model, the relevant guidelines, clinical decisions, evidence summaries, systematic reviews and expert consensus on non-pharmacological interventions for orthostatic hypotension in Parkinson disease patients were systematically retrieved from domestic and foreign databases. The search time was from the establishment of the database to December 2022. Two researchers independently screened the literature, evaluated the quality of the included literature, and extracted and summarized evidence that met the quality standards.Results:A total of 15 articles were included, including 4 guidelines, 2 clinical decision-making articles, 1 evidence summary article, 3 systematic evaluations and 5 expert consensus articles. A total of 24 pieces of best evidence were summarized from 7 aspects, including purpose, evaluation, capacity intervention, exercise intervention, posture intervention, physical intervention, health education and support.Conclusions:The best evidence on non-pharmacological intervention for orthostatic hypotension in patients with Parkinson disease can provide a reference for the practice of clinical medical staffs. It is suggested to apply the best evidence in combination with the patient's condition, preference and clinical environment, so as to reduce the incidence of orthostatic hypotension in patients with Parkinson disease and to ensure the safety of patients.

Objective:To investigate the value of high-risk HPV E6/E7 mRNA in situ hybridization in the grading of cervical squamous intraepithelial neoplasia (CIN).Methods:A total of 261 cases with diagnosis of CIN and cervical squamous cell carcinoma (SCC) at west China University Second Hospital, Sichuan University from July 2019 to June 2020 were collected, including 60 cases of CIN1, 41 cases of CIN2, 51 cases of CIN3, 72 cases of SCC, and 37 cases of normal cervical control tissue (10 HPV negative and 27 HPV positive). All pathologic tissues were made into tissue microarrays, and HE staining, HPV E6/E7 mRNA in situ hybridization (ISH) and p16 immunohistochemical (IHC) staining were performed respectively. The staining was assessed by light microscopy, and the positive rate and positive pattern were analyzed statistically.Results:HPV mRNA ISH in CIN1 mainly showed spot staining in predominantly basal to mid-epithelial layers (≤BME) with a diffuse nuclear signals in the superficial layer (supD), that is, the pattern of ≤BME+supD; in CIN2, it mainly showed spot staining in predominantly basal to above mid-epithelial but not the full layer (>BME) and some cases with supD staining, that is, the pattern of>BME+supD; In CIN3, the mainly pattern was >BME, and the spot staining was distributed throughout the epithelium. In CIN1, CIN2 and CIN3, there were significantly statistical differences among the above three staining patterns ( P<0.05). Conclusions:HPV mRNA ISH contributes to the accurate diagnosis and grading of CIN, and has better specificity than p16 IHC staining.

Objective:To explore the effect of the specialized nursing team model in children with severe asthma.Methods:A total of 80 children with severe asthma admitted to the Respiratory Intensive Care Unit, Tangshan Maternal and Child Health Hospital of Hebei Province from October 2018 to October 2019 were selected and divided into control group and observation group according to the random number table, with 40 cases in each group. Children in control group were given routine nursing care, and children in intervention group were treated with the specialized nursing team model, and they were all treated until discharge. We compared the efficacy, disease perception with the Brief Illness Perception Questionnaire (BIPQ) , medical phobia with the Children's Medical phobia Questionnaire, and quality of life one month after discharge with the Pediatric Asthma Quality of Life Questionnaire (PAQLQ) .Results:On the day of discharge, the total effective rate of clinical treatment of children in observation group was significantly higher than that of control group; and the scores of BIPQ and Children's Medical phobia Questionnaire of observation group were lower than those of control group, the differences were statistically significant ( P<0.05) . One month after discharge, the scores of symptom dimension and the total score of the PAQLQ of observation group were higher than those of control group, and the differences were statistically significant ( P<0.05) . Conclusions:The specialized nursing team model has a significant effect in the nursing care of severe asthma, which is conducive to improving the clinical efficacy of children, reducing children's disease perception and medical phobia, and improving children's quality of life.

Severe liver disease during pregnancy is uncommon in clinical practice. The most common cause of liver disease during pregnancy is liver dysfunction, with an overall prevalence rate of approximately 3%. Liver disease during pregnancy is classified into the liver diseases directly caused by pregnancy and those co-existing with pregnancy, i.e., pre-existing liver disease or occasional liver disease during pregnancy. A differential diagnosis of pre-existing and co-existing liver diseases may help to improve maternal and fetal outcome. During clinical diagnosis and selection of treatment and intervention measures, priority should be given to the potential impact on mother and fetus. This article introduces the latest research advances in the general information, pathogenesis, treatment, and pregnancy outcome of major liver diseases during pregnancy and elaborates on the risk of pregnancy and related coping measures for patients with pre-existing liver disease, so as to guide clinical diagnosis and treatment and patient management.

Objective: To study the predictive value of general movements(GMs) quality assessment technique(writhing movements)on the motor development outcome of high-risk infants, so as to provide a reference basis for clinical diagnosis and treatment. Methods: A retrospective analysis was made on the high-risk infants who were hospitalized in the Neonatal Department of the Affiliated Hospital of Inner Mongolia Medical University from January 1, 2017 to December 31, 2018, and the GMs quality assessment was finished and followed up to 12-month-old among high-risk infants.The clinical diagnostic criteria for patients with cerebral palsy and Peabody Development Motor Scales-2(PDMS-2)were used to evaluate the motor development outcome of 12-month-old high-risk infants.Furthermore, the predictive value of GMs writhing movements on the motor development outcome of high-risk infants were evaluated. Results: The predictive validity of writhing movements phase[cramped synchronized(CS)+ poor repertoire(PR)]for motor retardation and cerebral palsy in high-risk infants who met the inclusion criteria were as follows: the sensitivity, specificity, positive predictive value, negative predictive value were 94.44%, 23.03%, 11.04%, 97.62% and 100%, 21.88%, 2.60%, 100%, respectively.The predictive sensitivity and negative predictive value of writhing movements PR for motor retardation and cerebral palsy were 92.31%, 100%; 98.18%, 100% respectively.The predictive sensitivity, specificity and negative predictive value of writhing movements CS for motor retardation and cerebral palsy were 100%, 95.81%, 100% and 100%, 95.31% and 100%, respectively. Conclusion GMs quality assessment(writhing movements)has high reliability in predicting the motor development outcome of high-risk infants, especially cramped-synchronized has significant value in early screening of children with motor retardation and cerebral palsy.

Neonatal asphyxia is a clinical disease in which newborns are unable to establish regular and effective respiration after birth due to various factors, resulting in hypoxia, acidosis and hypercapnia.Severe cases can cause damage to the nervous system of children, mental retardation, cerebral palsy, and even death.The quality assessment of systemic exercise(general movements, GMs)is a simple, non-invasive and easy to implement technique for early diagnosis of neurodevelopmental disorders in children.Although GMs quality assessment has been widely used in the assessment of nervous system injury after asphyxia resuscitation at home and abroad, there has not been a systematic introduction to its pathogenesis, specific manifestations and the correlation between them.Therefore, this paper systematically reviewed the above problems combined with the latest research at home and abroad, in order to provide help for clinical work.