ObjectiveTo explore the protective effect and mechanism of Gegen Qianliantang （GQT） on intestinal mucosal barrier function in dextran sulfate sodium （DSS）-induced ulcerative colitis （UC） model mice. MethodsA UC model was established in C57BL/6 mice using a 2.5% DSS solution. Mice were randomly divided into five groups （n=8 per group）： blank group， model group， mesalazine sustained-release granule group （0.52 g·kg^-1）， high-dose GQT group （2.23 g·kg^-1）， and low-dose GQT group （1.12 g·kg^-1）. Fecal characteristics and body weight changes were observed before and after treatment. The body weight loss and disease activity index （DAI） of UC mice were calculated to evaluate symptom severity. Hematoxylin-eosin （HE） staining and Alizarin blue-periodic acid-Schiff （AB-PAS） staining were used to detect histological changes in colon tissue. Immunohistochemistry was used to detect the expression of zonula occludens-1 （ZO-1） and mucin 2 （MUC2）. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum levels of pro-inflammatory cytokines interferon-γ （IFN-γ）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， IL-17A， and anti-inflammatory cytokine IL-10. Flow cytometry was used to detect the activation of helper T lymphocyte subsets （Th1， Th17）， regulatory T cells （Treg）， and regulatory B cells （Breg） in spleen and colon tissues. Western blot was used to detect the expression levels of T-bet， forkhead box protein P3（FoxP3）， nuclear transcription factor（NF）-κB p65， phosphorylated NF-κB p65 （p-NF-κB p65）， signal transducer and activator of transcription 3（STAT3）， and phosphorylated STAT3 （p-STAT3）. ResultsCompared with the model group， both high- and low-dose GQT groups significantly improved the body weight loss and DAI scores （P<0.05）， alleviated colonic inflammation， and showed optimal efficacy in the high-dose group. AB-PAS staining showed that compared with the model group， both the high- and low-dose GQT groups significantly increased goblet cell proliferation and mucin secretion， indicating improved mucosal barrier function. GQT upregulated the expression of ZO-1 and MUC2 in colon tissue （P<0.05）， suppressed IFN-γ， IL-6， and TNF-α secretion （P<0.05）， elevated IL-10 secretion （P<0.05）， but had no significant effect on IL-17A. At the same time， high- and low-dose GQT intervention increased the activation of CD4⁺ FoxP3⁺ Treg cells （P<0.05） and suppressed activation of CD4⁺ IFN-γ⁺ Th1 cells （P<0.05）. Western blot showed that GQT downregulated T-bet， NF-κB p65， and STAT3 protein expression （P<0.05）， upregulated FoxP3 （P<0.05）， and also reduced phosphorylation levels of p-NF-κB p65 and p-STAT3 （P<0.05）. ConclusionGQT can upregulate the activation of CD4⁺ FoxP3⁺ Treg cells， reduce the activation of CD4⁺ IFN-γ⁺ Th1 cells， inhibit the secretion of IFN-γ， IL-6， and TNF-α， and increase the secretion of IL-10. It enhances the expression of MUC2 and ZO-1 in colon tissue， thereby alleviating inflammatory damage to the intestinal mucosa and restoring mucosal barrier integrity. These effects may be related to its regulation of NF-κB p65 and STAT3 signaling pathways， ultimately regulating the activation of transcription factors T-bet and FoxP3.

Bronchiectasis has the characteristics of long course,gradual aggravation,easy recurrence and difficult to treat.The characteristics are similar to those arouse by incubative pathogenic factors.Based on the theory of incubative pathogenic factors,this disease is often related to the incubative pathogenic factors in the body's areas with deficient healthy qi,which occur at regular times.The etiology can be external,congenital,or internal.Treatment should focus on different characteristics of incubative pathogenic factors.Attention should be paid to clearing and dispersing in external pathogenic factors,while attention should be paid to supporting and promoting healthy qi in congenital pathogenic factors,and do not forget to remove internal pathogenic factors.

Objective:To explore the curative efficacy of Dingke Decoction in the treatment of post infectious cough based on the the real-world clinical data.Methods:Retrospective cohort study was conducted. 245 patients with cough after infection in the hospital of Jingan Chinese Medical Hospital from July 2021 to July 2022 were set as study objects. The fact that whether the patients with Dingke Decoction or not were divided into control group (90 cases, without Dingke Decoction) and treatment group (155 cases, with Dingke Decoction). By using propensity nearest neighbor 1:1 matching to balance the confounding factors before treatment, 56 cases were successfully matched in both groups. The control group was treated symptomatically according to the actual clinical situation, while the treatment group was treated with modified Dingke Decoction on the basis of symptomatic treatment in the control group. The treatment for both groups lasted for 2 weeks. TCM symptom scores and cough severity score were evaluated using the Leicester Cough Quality of Life Questionnaire (LCQ) before and after treatment. Levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) were detected through ELISA; adverse reactions during the treatment were recorded, and the clinical efficacy was evaluated.Results:The total effective rate of the treatment group was 91.07% (51/56), while that of the control group was 76.79% (43/56), with a statistically significant difference between the two groups ( χ2=4.24, P=0.040). The daytime cough symptom score (1.03 ± 0.67 vs. 1.20 ± 0.66, t=7.40) and nighttime cough symptom score (0.74 ± 0.62 vs. 1.26 ± 0.54, t=6.27) in the treatment group were lower than those in the control group after treatment ( P<0.001). After treatment, the LCQ physiological (5.30 ± 0.79 vs. 4.78 ± 1.09, t=-2.44), psychological (5.33 ± 0.92 vs. 4.70 ± 1.12, t=-2.39), and social (5.23 ± 0.94 vs. 4.60 ± 0.81, t=-2.86) scores in the treatment group were higher than those in the control group ( P<0.05). After treatment, he treatment group serum IL-6 [(14.29 ± 3.94) ng/L vs. (19.99 ± 5.17) ng/L, t=4.80] and TNF-α [(36.23 ± 7.83) ng/L vs. (42.44 ± 7.63) ng/L, t=3.11] were lower than those in the control group ( P<0.01). The incidence of adverse reactions in the control group was 8.92% (5/56) and 5.36% (3/56), respectively, without statistical significance ( χ2=0.54, P=0.716). Conclusion:Based on the real-world research method, medicine Dingke Decoction can improve the clinical efficacy and the quality life of post infectious cough patients, and the mechanism may be related to reducing airway inflammation response.

Objective To evaluate the immunogenicity and protective effect of Mycobacterium tuberculosis(M.tb) Ag85B-Fc2a DNA vaccine in mice,so as to provide experimental basis for the development of the vaccine.Methods The recombinant plasmid pcD-Ag85B-Fc2a was identified by double digestion and sequencing,and then transfected into CHO-K1 cells.The expression of fusion protein Ag85B-Fc2a was detected by Western blot.Vector pcDNA3.1(+) and recombinant plasmid pcDAg85B-Fc2a were injected into female C57BL/6J mice through thigh muscle respectively(control group and immunization group),with 10 mice in each group,and booster immunization was carried out two weeks after the initial immunization.At14 d,28 d and 42 d after the initial immunization,serum samples was separated,and the titers of IgG antibody in the serum were detected by indirect ELISA.At 42 d after the initial immunization,the spleen of mice was taken aseptically and made into single cell suspension,and the proportions of CD4~+and CD8~+T cells were detected by flow cytometry.The remaining mice were injected with M.tb H37Ra through the tail vein 42 d after the initial immunization at a dose of 10~6 CFU/mouse.After 28 d of challenge,the lung and spleen of mice were collected aseptically.The number of bacteria in the left lung and spleen was measured by plate method,and the bacteria in the right lung was detected by auramine O fluorescence staining.Results Double digestion and sequencing results showed that the recombinant plasmid pcD-Ag85B-Fc2a was constructed correctly.After transfection into CHO-K1 cells,the fusion protein Ag85B-Fc2a with a relative molecular mass of about 70 000was detected.The Ag85B-specific IgG antibody titer in serum of mice in the immunization group was 1:3 200,1:12 160,and 1:12 800 at 14 d,28 d,and 42 d after the initial immunization,respectively,but no antibody titer was detected in the serum samples of control group.At 42 d after the initial immunization,the percentages of CD4~+ T cells in mouse spleen of control group and immunization group were(23.61±0.64)% and(26.92±0.80)%,and the percentages of CD8~+T cells were(14.12±0.87)% and(18.78±0.94)%,respectively,with significant differences(t=3.23 and 3.64,respectively,each P <0.05).After infection with M.tb H37Ra for 28 d,the numbers of bacteria were(4.73±0.13) and(3.81±0.14)CFU in the left lung,and(5.02±0.19) and(4.30±0.13) C.FU in the spleen of control group and immunization group,respectively,with significant differences(t=4.65 and 3.12,respectively,each P <0.01).The bacteria loading in the right lung was consistent with that in the left lung.Conclusion Ag85B-Fc2a DNA vaccine can induce specific humoral and cellular immune effects in mice,and can produce good protective effect against M.tb H37Ra infection.

Objective:To discuss the method for establishing the rat models of irritable bowel syndrome(IBS)by chronic water avoidance stress(WAS)method,and to evaluate its feasibility.Methods:Thirty male Wistar rats were randomly divided into control group(n=10)and model group(n=20).The rats in model group were induced by WAS method for 1 h everyday,lasting for 10 consecutive days;the rats in control group underwent no interventions.After modeling,the general conditions and body weights of the rats in two groups were observed and recorded.The elevated plus maze(EPM)test was used to detect the percentages of the number of open arm entries(OE)and the time spent in open arms(OT)of the rats in two groups;the abdominal withdrawal reflex(AWR)test was used to assess the visceral sensitivity of the rats in two groups;electrocardiography was used to detect the heart rate variability(HRV)of the rats in two groups;electromyography(EMG)of the external oblique muscle was used to detect the colorectal pain sensitivity thresholds of the rats in two groups;multi-channel physiological signal recorder was used to monitor the slow wave frequency of the colon of the rats in two groups.Results:There were no death rats in both groups during the modeling period.After modeling,the rats in model group exhibited poor mental status,reduced spontaneous activity,hypoactivity,disordered and dull fur,irritability,and unclean anal areas;whereas,the rats in control group showed no significant changes in the mental state,spontaneous activity,fur,and perianal area.Compared with control group,the body weight of the rats in model group was significantly decreased(P<0.05).The EPM test results showed that compared with control group,the OE percentage and OT percentage of the rats in model group were significantly decreased(P<0.01).The AWR test results showed that 12 rats in model group scored≥3 points,indicating that the successful rate in creating the visceral pain models was 60%.Compared with control group,the low frequency(LF)signals and the ratio of LF/high frequency(HF)of the rats in model group were significantly increased(P<0.01),and the HF was significantly decreased(P<0.05).The EMG results showed that compared with control group,the coloretal pain sensitivity threshold of the colon of the rats in model group was significantly decreased(P<0.01),and the slow wave frequency of the colon was significantly increased(P<0.01).Conclusion:The WAS method for establishing the rat model of IBS effectively demonstrates the changes in behavior and mental state,increased the visceral sensitivity,accelerated colonic slow wave frequency,and autonomic nervous system imbalance;the WAS method can serve as an effective modeling approach for observing and evaluating the related drugs and interventions on treatment of IBS.

Traditional Chinese Medicine (TCM) external therapy for sleep disorder of chronic fatigue syndrome (CFS) has good anti-fatigue effect and can improve sleep quality of patients. The treatment for sleep disorders of CFS with TCM external treatment mainly adopts acupuncture, moxibustion, massage, TCM bath, transcutaneous acupoint electrical stimulation and auricular point sticking, etc., or alone, or comprehensive application, or combined with oral Chinese materia medica. The appropriate treatment method can be selected according to the patients' condition and compliance, which reflects the unique advantages of TCM syndrome differentiation and treatment and the treatment according to people and time. The existing research still needs to further form a standardized and recognized diagnosis and treatment system, so as to better guide clinical popularization and application.

The treatment rules of point selection and treatment principles for treating chronic fatigue syndrome (CFS) can be divided into three categories: regulating and replenishing, invigorating original yang and regulating zang-fu organs. The mechanism of moxibustion includes improving gut microbiota imbalance, regulating immune cell imbalance and correcting endocrine dysfunction. The moxibustion methods include ginger-partitioned moxibustion, thunder-fire moxibustion, warm acupuncture, and governor moxibustion. Acupuncture points such as Shenque (RN8), Guanyuan (RN4), Qihai (RN6), Zusanli (ST36), Baihui (DU20), Yongquan (KI1) and back-shu points are often selected to exert anti-chronic fatigue effects.

Objective:To investigate the effect of CCN1 on the chemosensitivity of colon cancer cells to 5-FU .Methods:Colon cancer and adjacent tissues, colon cancer cells and normal colon epithelial cells, HCT-116 and HCT-116/5/FU cells were collected, and the SCD1 mRNA expression levels were detected by RT-qPCR; HCT-116 cells were cultured and transfected with pcDNA3.1 and CCN1 expression vectors, or infected with shNC and shCCN1 lentivirus, CCK-8 assay was used to detect cell sensitivity to 5-FU, Western blot and RT-qPCR were used to detect SCD1 mRNA expression, and oil red O staining was used to detect the lipid content. Western blot was used to detect the distribution of transcription factor FoxO1 in the nucleus and cytoplasm. The effect of CCN1 and FoxO1 on the transcriptional activity of SCD1 promoter was detected by luciferase assay.Results:Compared with control group, the expression of SCD1 was up-regulated in colon cancer tissues, cell lines and HCT-116/5-FU cells (all P<0.05); overexpression of CCN1 reduced the sensitivity to 5-FU, increased intracellular lipid deposition, and up-regulated the expression of SCD1 ( P<0.05); Knockdown of CCN1 increased the sensitivity to 5-FU, reduced intracellular lipid content and down-regulate the expression level of SCD1 ( P<0.05); CCN1 can promote FoxO1 nuclear distribution, activation or inhibition of FoxO1 activity can promote or up-regulate SCD1 expression level and promoter activity ( P<0.05). Conclusion:CCN1 may up-regulate the expression of SCD1 by activating FoxO1 activity and inhibit the sensitivity of colon cancer cells to 5-FU.

Objective: To investigate the effect of lncRNA GIHCG on the radiosensitivity of glioma cells and its mechanism. Methods: The expression levels of GIHCG and miR-146a-3p in human brain normal glial cells HEB and glioma cell lines U251, A172, SHG139 and U87 were quantitatively measured by qRT-PCR assay. U251 and SHG139 cells were used for subsequent experiment. After silencing the expression of GIHCG or overexpressing miR-146a-3p in U251 and SHG139 cells, cell proliferation was detected by MTT assay, cell apoptosis was detected by flow cytometry, cell radiosensitivity was detected by colony formation assay and the expression levels of CDK1, CyclinD1, Bcl-2 and Bax proteins were measured by Western blot. The bioinformatics software predicted the presence of a binding site for GIHCG and miR-146a-3p. Dual luciferase reporter gene assay and qRT-PCR assay were adopted to verify the targeting relationship between GIHCG and miR-146a-3p. Results: Compared with HEB cells, the expression of GIHCG was significantly up-regulated in glioma U87, U251, A172 and SHG139 cells (all P<0.05), whereas that of miR-146a-3p was remarkably down-regulated (P<0.05). Silencing GIHCG expression or overexpression of miR-146a-3p significantly decreased the U251 and SHG139 cell survival rate, survival fraction and the expression of CDK1, CyclinD1 and Bcl-2 proteins (all P<0.05), whereas considerably increased the apoptotic rate and expression of Bax protein (both P<0.05). GIHCG performed targeted negative regulation of miR-146a-3p expression in U251 and SHG139 cells and inhibition of miR-146a-3p expression reversed the effect of silencing GIHCG on proliferation, apoptosis and radiosensitivity of glioma cells. Conclusion Silencing GIHCG expression up-regulates the expression of miR-146a-3p, thereby enhancing the radiosensitivity of glioma cells.