Objective To establish an HPLC method to determine the concentration of inositol nicotinate（IN） in rat skin, and study the pharmacokinetic characteristics of IN after transdermal administration of heparin sodium inositol nicotinate cream in rats. Methods HPLC method was used to establish a simple and rapid analytical method for the determination of IN concentration in the skin of rats at different time points after administration. The established method was used to study the pharmacokinetics of IN after transdermal administration of heparin sodium inositol nicotinate cream in rats, and the pharmacokinetic parameters were fitted with DAS software. Results The linearity of the analytical method was good in the concentration range of 0.25-20 μg/ml, the quantitative limit was 0.25 μg/ml, and the average recovery rate was 96.18%. The pharmacokinetic parameters of IN after transdermal administration of heparin sodium inositol nicotinate cream in rats were as follows: t_1/2 was （4.555±2.054） h, T_max was （6±0）h, C_max was （16.929±2.153）mg/L, AUC_0−t was （150.665±16.568） mg·h /L ,AUC_0−∞ was （161.074±23.917） mg·h /L, MRT_（0−t） was （9.044±0.618）h, MRT_{（0−∞）} was （10.444±1.91） h, CLz/F was （0.19±0.03） L/（h·kg）, and Vz/F was （1.19±0.437） L/（h·kg）. Conclusion IN could quickly penetrate the skin and accumulate in the skin for a long time, which was beneficial to the pharmacological action of drugs on the lesion site for a long time. The method is simple, rapid, specific and reproducible, which could be successfully applied to the pharmacokinetic study of IN after transdermal administration in rats.

Object To study the efficacy and potential mechanism of Tongbianling capsule in constipation. Methods The effects of Tongbianling capsule on intestinal motility in normal mice and carbon powder propulsion rate in small intestine of constipation model mice after were observed administration. The potential targets and key pathways of Tongbianling capsule in treating constipation were identified through network pharmacology. To verify the mechanism, the expression of p-PI3K/PI3K, p-AKT/AKT and CASP3 proteins in mouse colon tissue was detected by the western blot. Results The time for mice to excrete the first black stool was shortened and the number of fecal particles was increased in Tongbianling capsule administration group, and the carbon powder propulsion rate of mice in each Tongbianling capsule administration group was increased. The results of network pharmacology showed that treatment of constipation by Tongbianling capsule may be related to signaling pathways such as PI3K-Akt signaling pathway and 5-HT. The protein expression of p-PI3K/PI3K, p-AKT/AKT, and CASP3 in mouse colon tissue could be significantly downregulated in administration group. Conclusion Tongbianling capsule could effectively promote intestinal peristalsis in mice, increase the frequency of defecation, and effectively treat constipation. The mechanism of its action may be related to the direct or indirect regulation of intestinal motility by the PI3K-Akt signaling pathway.

ObjectiveThis study aims to investigate the therapeutic effects of Wenweishu granule （WWSG） on functional dyspepsia （FD） with spleen-stomach deficiency cold syndrome in rats by integrating network pharmacology， molecular docking， and animal experiments. MethodsActive components and corresponding targets of WWSG were collected from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and the Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine （BATMAN-TCM）. Disease-related targets for FD with spleen-stomach deficiency cold syndrome were screened using GeneCards and the Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine （TCMIP）. Core therapeutic targets were identified via Cytoscape and validated by molecular docking. A rat model of FD with spleen-stomach deficiency cold syndrome was established using vinegar gavage combined with tail-clamping. The rats were randomly divided into a model group， low-， medium-， and high-dose WWSG groups （2.0， 4.0， 8.0 g·kg^-1）， a domperidone group （3.0 mg·kg^-1）， a Fuzi Lizhong pillwan （0.8 g·kg^-1）， and a normal control group （n=10 per group）. Drugs were administered once daily by gavage for 14 consecutive days. After treatment， body weight， symptom scores， and gastrointestinal motility indices were recorded. Gastric and duodenal pathologies changes were observed via hematoxylin-eosin （HE） staining. Brain-gut peptides were measured in serum and tissue using enzyme-linked immunosorbent assay （ELISA）. Immunohistochemistry and Western blot were performed to assess stem cell factor （SCF） and receptor tyrosine kinase （c-Kit） protein expression in gastric tissues. ResultsA total of 305 drug targets， 1 140 disease targets， and 116 overlapping targets were identified. Cytoscape analysis revealed 104 core targets. Enrichment analysis indicated that the SCF/c-Kit signaling pathway was the key mechanism. Molecular docking confirmed a strong binding affinity between active components of WWSG and SCF/c-Kit proteins （binding energy<-5.1 kcal·mol^-1）. Compared with the normal group， model rats exhibited slower weight gain （P<0.05）， reduced gastric emptying and intestinal propulsion （P<0.01）， mild gastric mucosal shedding， duodenal inflammatory cell infiltration， decreased levels of gastrin （GAS）， 5-hydroxytryptamine （5-HT）， and vasoactive intestinal peptide （VIP） （P<0.05， P<0.01）， and elevated somatostatin （SS） expression （P<0.05， P<0.01）. WWSG treatment ameliorated weight gain， symptom scores， and low-grade inflammation in gastric/duodenal tissues. High-dose WWSG significantly improved gastric emptying and intestinal propulsion， upregulated GAS， 5-HT， and VIP， and downregulated SS expression in serum and tissues （P<0.05， P<0.01）. Immunohistochemistry and Western blot demonstrated that SCF and c-Kit protein expression was decreased in the model group （P<0.05， P<0.01）， which was reversed by WWSG intervention （P<0.05）. ConclusionWWSG exerts therapeutic effects on FD with spleen-stomach deficiency cold syndrome in rats， potentially by regulating the SCF/c-Kit signaling pathway to enhance gastrointestinal motility.

ObjectiveTo investigate the effects of Huashi Runzao prescription （HRP） on the histopathological injury and function of submandibular gland in naive non-obese diabetic （NOD/Ltj） mouse model of Sjögren's syndrome （SS） and its regulatory effect on aquaporin 5 （AQP5） expression in submandibular gland cells. MethodThe SS model was induced in NOD/Ltj mice. The NOD/Ltj female mice aged nine weeks were selected and randomly assigned into model group，HRP group （7.15 g·kg^-1·d^-1），and hydroxychloroquine （HCQ） group （1.30 g·kg^-1·d^-1）， and female BALB/c mice in the same age were selected and assigned into the normal group， with six mice in each group. Drug intervention lasted eight weeks. The water consumption and salivary flow rate （SFR） of each group were recorded. The pathological staining results of the submandibular gland of mice in each group were observed and scored. AQP5 expression was determined by immunohistochemistry （IHC） and Western blot. ResultCompared with the normal group， the model group showed increased water consumption （P<0.05） and reduced SFR （P<0.05）. Compared with the model group， the HRP group showed decreased water consumption （P<0.05） and increased SFR （P<0.05）， and the HCQ group showed increased SFR （P<0.05）. In terms of histopathological results of the submandibular gland，compared with the normal group，the model group showed increased pathological score， number of lymphocyte infiltration foci，and percentage of lymphatic infiltration area （P<0.05）. Compared with the model group， the HRP group showed reduced pathological scores and number of lymphocyte infiltration foci （P<0.05）， and the HRP group and the HCQ group showed reduced percentage of lymphatic infiltration area（P<0.05）. The results of IHC and Western blot showed that compared with the normal group，the model group showed down-regulated expression level of AQP5 protein （P<0.05）， and compared with the model group and the HCQ group，the HRP group showed up-regulated expression level of AQP5 protein （P<0.05）. ConclusionHRP can improve the secretion function of submandibular gland acinous cells and glandular structure injury in SS model mice， and its mechanism may be related to the up-regulation of AQP5 protein expression level in submandibular gland cells.

ObjectiveTo observe the efficacy and safety of Huashi Runzao prescription for patients with primary Sjögren's syndrome （pSS） of combined dryness and dampness pattern. MethodA total of 105 eligible patients were randomized into the experimental group （65 cases） and control group （40 cases）， and they were respectively treated with Huashi Runzao prescription and hydroxychloroquine for 12 weeks. Visual Analogue Scale （VAS） was employed to assess the symptoms. The symptoms of dryness， fatigue， and pain， European League Against Rheumatism （EULAR） Sjögren's Syndrome Patient Reported Index （ESSPRI）， EULAR Sjögren's syndrome disease activity index （ESSDAI）， and immune inflammatory indicators before and after treatment were compared between the two groups， and adverse reactions were observed. ResultAfter treatment， the ESSPRI score was lower than that before treatment in the experimental groups （P<0.01） and was lower in the experimental group than in the control group （P<0.05）. The VAS scores of dry mouth， dry eyes， overall dryness， fatigue， and pain in the experimental group decreased compared with those before treatment （P<0.01）， and the experimental group had lower VAS scores of dry mouth and overall dryness than the control group （P<0.01）. After treatment， the ESSDAI score of both groups decreased compared with that before treatment （P<0.05， P<0.01）， but there was no significant difference between the groups. After treatment， the level of immunoglobulin M （IgM） decreased （P<0.01） and the level of complement C3 increased （P<0.01） in the experimental group， while the level of complement C3 decreased in the control group （P<0.05）. There was no significant difference in the laboratory indexes between groups. During the treatment， stomachache occurred to one case in the experimental group， which was alleviated after the treatment， and no adverse reaction was observed in the control group. According to the chi-square test， the occurrence of adverse reactions was insignificantly different between the two groups. ConclusionHuashi Runzao prescription can alleviate the symptoms of dryness， fatigue， and pain， and reduce disease activity without associated side effects in pSS patients with combined dampness and dryness pattern.

ObjectiveTo explore the characteristics of tongue manifestations and traditional Chinese medicine （TCM） syndromes in patients with primary Sjogren's syndrome （pSS） of different time. MethodpSS patients who visited TCM Department of Rheumatism in China-Japan Friendship Hospital from December 2018 to October 2021 were included and grouped according to the disease courses （short<5 years， medium 5-10 years， long > 10 years）. Chi-square test was used for comparison between three groups and Bonferroni method for multiple comparisons. In the pairwise comparison， P<0.017 indicated significant difference. Chi-square test was performed on the syndrome and specific tongue manifestations with inter-group differences to analyze the trend of tongue manifestations and TCM syndromes over time （interval： two years）. ResultA total of 193 pSS patients were enrolled， with 85 （44.0%） of short disease course， 69 （35.8%） of medium disease course， and 39 （20.2%） of long disease course. The common tongue manifestations were crimson tongue， fissured tongue， thin tongue， lack of fluid， and dry coating， which showed no significant difference among the three groups. Higher proportion of patients with light red tongue was observed in the group with short disease course than in group with medium disease course （χ²=6.407， P<0.017）. Higher proportions of patients with thick coating （χ²=6.784， P<0.017） and phlegm-dampness syndrome （χ²=11.545， P<0.017） and lower proportion of patients with Qi deficiency syndrome （χ²=12.706， P<0.017） were found in the group with short disease course than in the group with long disease course. Patients with medium （χ²=6.358， P<0.017） and long （χ²=8.279， P<0.017） disease course tended to have exfoliated coating compared with those with short disease course， and the proportion of patients with exfoliated coating rose and the proportion of patients with thick greasy coating decreased over time （P_trend<0.05）. In addition， the proportion of patients with phlegm-dampness syndrome decreased and that with Qi deficiency syndrome increased over time （P_trend<0.05）. ConclusionIn the early stage， patients with pSS often show both dryness and dampness， as manifested by the thick greasy coating and phlegm-dampness syndrome. In the medium and late stage， patients often have Qi-Yin deficiency， as evidenced by exfoliated coating and Qi deficiency. In the clinical practice， medicines should be prescribed based on tongue manifestations and TCM syndrome of patients.

ObjectiveTo analyze the clinical characteristics of geographic tongue in patients with primary Sjögren's syndrome （pSS）. MethodPatients with pSS treated in the China-Japan Friendship Hospital from December 2018 to October 2021 were enrolled and divided into different groups according to the presence of geographic tongue or the severity. Phi correlation analysis，Chi-square test， and Mann-Whitney test were used to analyze the clinical characteristics of patients with geographic tongue. ResultA total of 182 pSS patients were included in this study，including 75 （41.2%） patients with geographic tongue and 107 （58.8%） without geographic tongue. Partial exfoliation was more common than total exfoliation， and the anterior part of the tongue was the most common exfoliation site. The number of patients with severe geographic tongue was more than those with mild-to-moderate geographic tongue. Compared with pSS patients without geographic tongue，those with geographic tongue were more common in fissured tongue （92.0%/41.1%，χ²=48.491，P<0.05），red or crimson tongue （48.0%/23.3%，χ²=12.009，P<0.05），Yin deficiency syndrome （100.0%/43.9%，χ²=62.739，P<0.05），and Qi deficiency syndrome （94.7%∶50.5%，χ²=40.046，P<0.05），less common in phlegm-dampness syndrome （33.3%/72.0%，χ²=26.709，P<0.05），and showed higher proportions in hyperglobulinemia （89.3%/65.4%，χ²=13.547，P<0.01），ANA ≥1∶160 （78.1%/57.3%，χ²=8.227，P<0.01），and positive RF （51.4%/36.5%，χ²=3.877，P<0.05）. Compared with pSS patients with mild geographic tongue，pSS patients with moderate-to-severe geographic tongue had higher proportions in hyperglobulinemia （98.1%/68.2%，χ²=14.617，P<0.01），positive anti-CENP-B （26.0%/4.8%，χ²=4.214，P<0.05），and reduced complement 3 （26.4%/4.5%，χ²=4.647，P<0.05）. The geographic tongue was positively associated with fissured tongue （φ=0.531），Yin deficiency syndrome （φ=0.587），and Qi deficiency syndrome （φ=0.469），negatively associated with phlegm-dampness syndrome （φ=-0.447），and weakly associated with tongue color （φ<0.4）. There was no statistical difference in the disease activity index between patients with or without geographic tongue and severity. ConclusionMore than 40% of pSS patients had geographic tongue accompanied by fissured tongue. Geographic tongue is positively associated with Yin deficiency syndrome and Qi deficiency syndrome，and negatively associated with phlegm-dampness syndrome， indicating that treatment should be based on tonifying Qi and nourishing Yin. Compared with pSS patients without geographic tongue， those with geographic tongue may have higher positive rate of some immune indicators，which deserves further exploration.

ObjectiveTo summarize the characteristics of traditional Chinese medicine （TCM） syndrome in primary Sjögren's syndrome （pSS） patients with interstitial lung disease （ILD） and to explore associated factors. MethodA survey was conducted and pSS patients who were treated in TCM department of rheumatism at China-Japan Friendship Hospital from December 2018 to April 2022 were included. Tongue manifestations and syndromes of patients were recorded. pSS patients with ILD were classified into the pSS-ILD group and those without the ILD were included in the pSS-non-ILD group. The tongue manifestations， syndromes， and laboratory indexes were compared between the two groups， and logistic regression was used to explore the factors associated with pSS-ILD. ResultA total of 200 pSS patients were included， with 186 （93.0%） females， median age of 57 years， and median disease course of 60 months， of which 44 （22%） had pSS-ILD. In terms of tongue manifestations， pSS-ILD patients generally had dark/purple/stasis tongue， fissured tongue， and tongue with little fluid， thick coating， yellow coating， and greasy coating. The proportion patients with yellow coating was higher in pSS-ILD group than in the pSS-non-ILD group （χ²=4.799，P<0.05）. In terms of syndrome， more than 40% of pSS-ILD patients had Qi deficiency， Yin deficiency， phlegm-dampness， Qi stagnation， and/or blood stasis syndrome. As for Yin deficiency， liver-kidney Yin deficiency syndrome ranked the first. For Qi deficiency， lung Qi deficiency syndrome was most commonly seen. The proportion of patients with lung Qi deficiency was higher in the pSS-ILD group than in the pSS-non-ILD group （χ²=18.667，P<0.01）. As to laboratory indexes， compared with the pSS-non-ILD group， pSS-ILD group had high proportion of anti-SSA-positive patients （P<0.05） and high levels of C-reactive protein （CRP） （P<0.01）， complement C3 （χ²=4.332，P<0.05）， and complement C4 （P<0.05）. Logistic regression analysis showed that pSS with ILD was positively associated with lung Qi deficiency ［odds ratio （OR）=6.079， 95% confidence interval （CI） 2.585-14.298， P<0.01）］ and yellow coating （OR=5.260， 95% CI 1.337-20.692， P<0.05） and negatively associated with low C4 （OR=0.199， 95% CI 0.070-0.564， P<0.01）. ConclusionAbout 22% of pSS patients had ILD， and patients with pSS-ILD generally have Qi deficiency， Yin deficiency， phlegm-dampness， Qi stagnation， and/or blood stasis syndrome. Yellow coating， lung Qi deficiency and C4 level are factors associated with pSS combined with ILD.

Objective:To explore the effectiveness of the artificial intelligence-endoscopic ultrasound (AI-EUS) biliary and pancreatic recognition system in assisting the recognition of EUS images.Methods:Subjects who received EUS due to suspicious biliary and pancreatic diseases from December 2019 to August 2020 were prospectively collected from the database of Department of Gastroenterology, Renmin Hospital of Wuhan University. Pancreatic EUS images of 28 subjects were included for recognition of pancreas standard station. EUS images of bile duct of 29 subjects were included for recognition of bile duct standard station. Eight new endoscopists from the Gastroenterology Department of Renmin Hospital of Wuhan University read the 57 EUS videos with and without the assistance of AI-EUS biliary and pancreatic recognition system. Accuracy of endoscopists' identification of biliary and pancreatic standard sites with and without the assistance of AI-EUS was compared.Results:The accuracy of pancreas standard station identification of the new endoscopists increased from 67.2% (903/1 344) to 78.4% (1 054/1 344) with the assistance of AI-EUS. The accuracy of bile duct standard station identification increased from 56.4% (523/928) to 73.8% (685/928).Conclusion:AI-EUS biliary and pancreatic recognition system can improve the accuracy of EUS images recognition of biliary and pancreatic system, which can assist diagnosis in clinical work.

Objective:To construct a deep learning-based diagnostic system for gastrointestinal submucosal tumor (SMT) under endoscopic ultrasonography (EUS), so as to help endoscopists diagnose SMT.Methods:From January 1, 2019 to December 15, 2021, at the Digestive Endoscopy Center of Renmin Hospital of Wuhan University, 245 patients with SMT confirmed by pathological diagnosis who underwent EUS and endoscopic submucosal dissection were enrolled. A total of 3 400 EUS images were collected. Among the images, 2 722 EUS images were used for training of lesion segmentation model, while 2 209 EUS images were used for training of stromal tumor and leiomyoma classification model; 283 and 191 images were selected as independent test sets to evaluate lesion segmentation model and classification model, respectively. Thirty images were selected as an independent data set for human-machine competition to compare the lesion classification accuracy between lesion classification models and 6 endoscopists. The performance of the segmentation model was evaluated by indexes such as Intersection-over-Union and Dice coefficient. The performance of the classification model was evaluated by accuracy. Chi-square test was used for statistical analysis.Results:The average Intersection-over-Union and Dice coefficient of lesion segmentation model were 0.754 and 0.835, respectively, and the accuracy, recall and F1 score were 95.2%, 98.9% and 97.0%, respectively. Based on the lesion segmentation, the accuracy of classification model increased from 70.2% to 92.1%. The results of human-machine competition showed that the accuracy of classification model in differential diagnosis of stromal tumor and leiomyoma was 86.7% (26/30), which was superior to that of 4 out of the 6 endoscopists(56.7%, 17/30; 56.7%, 17/30; 53.3%, 16/30; 60.0%, 18/30; respectively), and the differences were statistically significant( χ2=7.11, 7.36, 8.10, 6.13; all P<0.05). There was no significant difference between the accuracy of the other 2 endoscopists(76.7%, 23/30; 73.3%, 22/30; respectively) and model(both P<0.05). Conclusion:This system could be used for the auxiliary diagnosis of SMT under ultrasonic endoscope in the future, and to provide a powerful evidence for the selection of subsequent treatment decisions.