Background/Aims: The pathophysiology of refractory gastroesophageal reflux disease (RGERD), which differs from proton pump inhibitor dependent gastroesophageal reflux disease (DGERD), remains incompletely elucidated. This study aims to compare esophageal motility patterns, transdiaphragmatic pressure gradients (TPG), and reflux profiles between RGERD and DGERD patients, and to delineate dynamic pressure gradient-esophagogastric junction (EGJ) interactions in these patients. Methods: In this retrospective study, 274 patients who underwent 24-hour impedance-pH monitoring and high-resolution manometry, along with an assessment of proton pump inhibitor responsiveness, were classified as RGERD (32.5%), DGERD (54.4%), or non-GERD (13.1%). Clinical characteristics, TPG, esophageal motility, and reflux metrics were compared between RGERD and DGERD patients. Subgroup analysis excluding hiatal hernia (HH) was conducted to investigate the pathophysiology of RGERD. Results: The RGERD group exhibited a significantly higher proportion of chest pain compared to the DGERD group. Regarding reflux profiles, RGERD patients without HH (RGERDHH- group) experienced increased weakly acidic reflux (P < 0.001) and prolonged bolus exposure (P = 0.006) compared to their counterparts (DGERDHH- group). Mechanistically, the RGERDHH- group showed reduced lower esophageal sphincter basal pressure (P = 0.010) and EGJ contractile integral (P = 0.005). Notably, following a wet-swallow, the RGERDHH- group experienced the significant elevation in gastric pressure and TPG. Correlation analyses revealed weakly acidic reflux and bolus exposure were positively correlated with gastric pressure variation, and inversely correlated with lower esophageal sphincter basal pressure. Conclusions Transient gastric pressure elevation and compromised EGJ barrier function drive weakly acidic reflux and esophageal bolus exposure. This pressure gradient-barrier mismatch underpins the refractoriness of RGERD.

Objective:To explore the role of methyltransferase-like 3 (METTL3) on endometrial decidualization.Methods:Bioinformatics was used to analyze the expression of 26 N6-methyladenosine (m6A) regulators in recurrent implantation failure (RIF), recurrent spontaneous abortion (RSA) and the decidualization of endometrial stromal cells. Using Medroxyprogesterone and Dibutyryl-cAMP to induce human endometrial stromal cell s (HESCs)decidualization, we examined changes in the expression of total m6A, METTL3, Forkhead framing protein O1 (FOXO1), and decidualization markers insulin-like growth factor binding protein 1 (IGFBP1) and prolactin in the control and decidualization group. Constructing HESC models with METTL3 interference using lentivirus, we used CCK-8 and Edu to detect proliferation in the negative control and knockdown METTL3 group. Lentiviral transfection was successfully followed by induction of decidualization, and we examined changes in the expression of FOXO1 and decidualization markers.Results:The results of bioinformatics analysis showed that the expression of METTL3 was significantly downregulated in the endometrium of patients with RIF and RSA ( P=0.014, P=0.016), while it was significantly upregulated in the endometrium with decidualization ( P=0.029). In vitro, induction of HESCs decidualization increased in the expression levels of total m6A, METTL3, and FOXO1 proteins in the decidualization group ( P=0.015, P=0.016, P=0.004). Knocking down METTL3 resulted in a decrease in FOXO1 protein expression in HESCs ( P=0.009). The CCK-8 and Edu results showed that the proliferation of HESCs was inhibited after knocking down METTL3. After inducing the knockdown of METTL3 in vitro, there was no statistically significant difference in the expression levels of IGFBP1 mRNA, PRL mRNA, and FOXO1 protein in HESCs compared with the negative control group (all P>0.05). Conclusion:In endometrial stromal cells, METTL3 can regulate the expression of FOXO1, promote the expression of IGFBP1 and PRL, and affect the decidualization of endometrial cells.

Objective:To investigate the changes in follicular helper T cells(Tfh)and related cytokines in pe-ripheral blood and endometrial tissue of patients with unexplained recurrent implantation failure(URIF),and evalu-ate their predictive value for URIF.Methods:Sixty-four patients with URIF who visited the Reproductive Medicine Centre of the First Affiliated Hospital of Xinjiang Medical University from December 2020 to June 2022 were select-ed as the study group(URIF group),and 61 healthy women of childbearing age who visited our centre for male in-fertility in the same period were selected as the healthy control group(HC group).GSE microarrays of patients with recurrent implant failure(RIF)were collected in the Gene Expression Omnibus(GEO)database,and the pro-portion of different immune cells was identified using the CIBERSORT algorithm,and immune cell infiltration analy-sis and visualisation processing were performed using the Cibersort.R package.Peripheral blood samples and endometrial tissue samples were collected from the two groups of patients,flow cytometry was used to detect the proportion of Tfh cells in peripheral blood,enzyme-linked immunosorbent assay(ELISA)was used to detect serum levels of IL-4,IL-6,and IL-21,and immunohistochemical staining was used to detect the expression of CXCR5,Bcl-6,and IL-21 in endometrial tissue.Factors associated with URIF were analysed using multifactorial Logistic re-gression analysis.The predictive value of individual and combined tests for each indicator for URIF was analysed using the operating characteristic curve(ROC).The Drug Therapy Target Database(TTD)was used to predict potential therapeutic agents for URIF targeting IL-6 and IL-21,respectively.Results:The results of immune cell in-filtration in the GSE dataset in the GEO database showed that compared with normal controls,the numbers of ac-tivated memory CD4+T cells,Tfh,regulatory T cells(Treg),inactive macrophages,and resting dendritic cells were increased in the endometrial tissues of the patients with RIF(P＜0.05).The number of initial B cells,yδT cells,M2-type macrophages and activated dendritic cells were all decreased(P＜0.05).The levels of peripheral blood Tfh,IL-6 and IL-21 in the URIF group were all significantly higher than those in the HC group,and the difference was statistically significant(P＜0.05).Immunohistochemical staining results suggested that the expression of CX-CR5,Bcl-6 and IL-21 in the endometrium of patients in the URIF group was increased compared with that in the HC group(P＜0.05).Multivariate Logistic regression analysis showed that IL-6,IL-2,and Tfh were independent risk factors for the occurrence of URIF(OR＞1,P＜0.05).The area under the curve(AUC=0.974)of peripheral blood Tfh combined with IL-6 and IL-21 for diagnosis of URIF was higher than that predicted by each index alone,according to ROC analysis(P＜0.05).Five drugs targeting IL-6 or IL-21 with potential therapeutic effects on URIF were detected and screened through the TTD database.Conclusions:The levels of Tfh,IL-6 and IL-21 in the pe-ripheral blood of patients with URIF were abnormally elevated,and CXCR5,Bcl-6 and IL-21 were abnormally ex-pressed in the endometrium of patients with URIF,suggesting that Tfh cells and their cytokines are closely related to the occurrence and development of URIF.The combined prediction of these indicators has good diagnostic val-ue and may serve as a therapeutic target.

ObjectiveTo observe the apoptosis induced by paeoniflorin （PF） in non-small cell lung cancer （NSCLC） cells and explore its mechanism. MethodCell counting kit-8 （CCK-8） was used to detect the inhibition rates of H1299， H292 and A549 cells with different concentrations of PF （2.5， 5， 10， 20， 25 µmol·L^-1）， and to screen suitable concentrations of PF and experimental cells. The inhibitory effect of PF on lung cancer cells was detected by clone formation assay. The effect of PF on cell apoptosis was detected by flow cytometry with annexin V-FITC/propidium iodide （PI） double staining. With the right concentration of drugs， levels of apoptosis-associated protein B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， cleaved Caspase-3 and Caspase-3 were detected by Western blot. At the same time， the molecular expressions of hypoxia inducible factor -1α （HIF-1α） and Hippo signaling pathway were determined. ResultCompared with the blank group， PF significantly inhibited the growth of H1299， H292 and A549 cells of human lung cancer （P<0.01）. PF significantly induced apoptosis in A549 cells （P<0.01）， decreased the Bcl-2/Bax ratio （P<0.01）， and significantly increased the cleaved Caspase-3 expression （P<0.01）. Compared with those in the blank group， the expression levels of HIF-1α， transcriptional coactivator with PDZ-binding motif （TAZ）， large tumor suppressor 1 （LATS1）， Mps one binding 1 （MOB1） and Yes-associated protein （YAP） in A549 cells of the PF treatment group were significantly decreased （P<0.01）， while the expressions of p-LATS1， p-MOB1 and p-YAP were significantly increased （P<0.01）. At the same time， there was no significant effect on the expression levels of phosphorylated mammalian Ste20-like kinase 1 （p-MST1） and MST1， which did not reach a statistical difference. ConclusionAll data demonstrated that PF showed an anti-tumor effect by improving hypoxic conditions and inhibiting the abnormally activated Hippo signaling pathway， thereby inducing and promoting apoptosis in non-small cell lung cancer.

Objective To investigate the effect and mechanism of long chain non-coding RNA(ln-cRNA)SNHG16 regulating PAR1 on the proliferation,migration and invasion of lung cancer.Methods From March 2020 to August 2021,lung cancer tissues and adjacent tissues of 35 patients with lung cancer were col-lected,and lung cancer cell lines(HCC827,A549,SK-LU-1,A427)and normal lung cell lines(MRC5)were simultaneously cultured.The overexpression model of PAR1(pcDNA-PAR1)was constructed by using the expression vector pcDNA3.1.A549 cells were divided into four groups after transfection:si-SNHG16,si-PAR1,si-SNHG16+pcDNA-PAR1 and control(transfection with si-NC).The expression levels of lncRNA SNHG16 and PAR1 in lung cancer cell lines(HCC827,A549,SK-LU-1,A427),lung cancer tissues and adja-cent tissues were detected by real-time fluorescence quantitative reverse transcription-polymerase chain reac-tion(qRT-PCR),and the transfection efficiency of each group was verified.MTT assay and clonal formation were used to determine the proliferation of cells in each group,flow cytometry was used to detect the apopto-sis of cells in each group,cell scratch and Transwell test were used to detect the migration and invasion ability of cells in each group,and Western blot was used to detect the protein expression of PAR1.Results The ex-pression level of lncRNA SNHG16 in lung cancer tissue was higher than that in adjacent tissues(P＜0.05).The expression level of lncRNA SNHG16 in lung cancer cell lines(HCC827,A549,SK-LU-1,A427)was higher than that in normal lung cell lines(MRC5),with statistical significance(P＜0.05).The results of qRT-PCR showed that the expression level of lncRNA SNHG16 gene was(21.02±0.04)％of the control af-ter transfection of si-SNHG16,the expression level of PAR1 gene was(19.06±0.02)％of the control after transfection of si-PAR1,and the expression level of PAR1 gene was 2.70±0.00 folds of the control after transfection of pcDNA-PAR 1,the difference was statistically significant(P＜0.05).The results of Western blot showed that the expression level of transfected in each PAR1 protein was different from that of the con-trol(P＜0.05).Compared with the control,the activity of cells transfected with si-SNHG16 was lower,the a-bility of clone formation,cell migration and invasion was obviously inhibited,and the apoptosis rate was higher(P＜0.05),while pcDNA-PAR1 could weaken the influence of transfected si-SNHG16 on cell proliferation,apoptosis,migration and invasion(P＜0.05).lncRNA SNHG16 was positively correlated with the expression level of PAR1(r=0.61).Conclusion lncRNA SNHG16 can regulate the occurrence and development of lung cancer by targeting PAR1.

Objective:To explore the role of methyltransferase-like 3 (METTL3) on endometrial decidualization.Methods:Bioinformatics was used to analyze the expression of 26 N6-methyladenosine (m6A) regulators in recurrent implantation failure (RIF), recurrent spontaneous abortion (RSA) and the decidualization of endometrial stromal cells. Using Medroxyprogesterone and Dibutyryl-cAMP to induce human endometrial stromal cell s (HESCs)decidualization, we examined changes in the expression of total m6A, METTL3, Forkhead framing protein O1 (FOXO1), and decidualization markers insulin-like growth factor binding protein 1 (IGFBP1) and prolactin in the control and decidualization group. Constructing HESC models with METTL3 interference using lentivirus, we used CCK-8 and Edu to detect proliferation in the negative control and knockdown METTL3 group. Lentiviral transfection was successfully followed by induction of decidualization, and we examined changes in the expression of FOXO1 and decidualization markers.Results:The results of bioinformatics analysis showed that the expression of METTL3 was significantly downregulated in the endometrium of patients with RIF and RSA ( P=0.014, P=0.016), while it was significantly upregulated in the endometrium with decidualization ( P=0.029). In vitro, induction of HESCs decidualization increased in the expression levels of total m6A, METTL3, and FOXO1 proteins in the decidualization group ( P=0.015, P=0.016, P=0.004). Knocking down METTL3 resulted in a decrease in FOXO1 protein expression in HESCs ( P=0.009). The CCK-8 and Edu results showed that the proliferation of HESCs was inhibited after knocking down METTL3. After inducing the knockdown of METTL3 in vitro, there was no statistically significant difference in the expression levels of IGFBP1 mRNA, PRL mRNA, and FOXO1 protein in HESCs compared with the negative control group (all P>0.05). Conclusion:In endometrial stromal cells, METTL3 can regulate the expression of FOXO1, promote the expression of IGFBP1 and PRL, and affect the decidualization of endometrial cells.

Objective:To investigate the changes in follicular helper T cells(Tfh)and related cytokines in pe-ripheral blood and endometrial tissue of patients with unexplained recurrent implantation failure(URIF),and evalu-ate their predictive value for URIF.Methods:Sixty-four patients with URIF who visited the Reproductive Medicine Centre of the First Affiliated Hospital of Xinjiang Medical University from December 2020 to June 2022 were select-ed as the study group(URIF group),and 61 healthy women of childbearing age who visited our centre for male in-fertility in the same period were selected as the healthy control group(HC group).GSE microarrays of patients with recurrent implant failure(RIF)were collected in the Gene Expression Omnibus(GEO)database,and the pro-portion of different immune cells was identified using the CIBERSORT algorithm,and immune cell infiltration analy-sis and visualisation processing were performed using the Cibersort.R package.Peripheral blood samples and endometrial tissue samples were collected from the two groups of patients,flow cytometry was used to detect the proportion of Tfh cells in peripheral blood,enzyme-linked immunosorbent assay(ELISA)was used to detect serum levels of IL-4,IL-6,and IL-21,and immunohistochemical staining was used to detect the expression of CXCR5,Bcl-6,and IL-21 in endometrial tissue.Factors associated with URIF were analysed using multifactorial Logistic re-gression analysis.The predictive value of individual and combined tests for each indicator for URIF was analysed using the operating characteristic curve(ROC).The Drug Therapy Target Database(TTD)was used to predict potential therapeutic agents for URIF targeting IL-6 and IL-21,respectively.Results:The results of immune cell in-filtration in the GSE dataset in the GEO database showed that compared with normal controls,the numbers of ac-tivated memory CD4+T cells,Tfh,regulatory T cells(Treg),inactive macrophages,and resting dendritic cells were increased in the endometrial tissues of the patients with RIF(P＜0.05).The number of initial B cells,yδT cells,M2-type macrophages and activated dendritic cells were all decreased(P＜0.05).The levels of peripheral blood Tfh,IL-6 and IL-21 in the URIF group were all significantly higher than those in the HC group,and the difference was statistically significant(P＜0.05).Immunohistochemical staining results suggested that the expression of CX-CR5,Bcl-6 and IL-21 in the endometrium of patients in the URIF group was increased compared with that in the HC group(P＜0.05).Multivariate Logistic regression analysis showed that IL-6,IL-2,and Tfh were independent risk factors for the occurrence of URIF(OR＞1,P＜0.05).The area under the curve(AUC=0.974)of peripheral blood Tfh combined with IL-6 and IL-21 for diagnosis of URIF was higher than that predicted by each index alone,according to ROC analysis(P＜0.05).Five drugs targeting IL-6 or IL-21 with potential therapeutic effects on URIF were detected and screened through the TTD database.Conclusions:The levels of Tfh,IL-6 and IL-21 in the pe-ripheral blood of patients with URIF were abnormally elevated,and CXCR5,Bcl-6 and IL-21 were abnormally ex-pressed in the endometrium of patients with URIF,suggesting that Tfh cells and their cytokines are closely related to the occurrence and development of URIF.The combined prediction of these indicators has good diagnostic val-ue and may serve as a therapeutic target.

Objective:To investigate the preventive effect of Xiyanping injection on radioactive esophagitis in patients with radical radiotherapy and chemotherapy for esophageal cancer.Methods:A total of 70 patients with esophageal cancer undergoing radical radiotherapy and chemotherapy were selected from the Department of Radiation Oncology of Jiangsu Provincial People′s Hospital from January to September 2020. They were divided into experimental group ( n=35) and control group ( n=35) according to random number table method. The control group received standard concurrent radiotherapy and chemotherapy, and the experimental group received concurrent radiotherapy and chemotherapy combined with Xiyanping. The white blood cell count, neutrophil count, procalcitonin (PCT) and interleukin-6 (IL-6) levels before and after treatment were compared between the two groups, as well as the occurrences of radioactive esophagitis and radioactive pneumonia during radiotherapy. Results:Before treatment, there were no significant differences in white blood cell count [4.57 (2.52)×10 9/L vs. 5.59 (2.23)×10 9/L] and neutrophil count [2.95 (1.66)×10 9/L vs. 3.69 (1.56)×10 9/L] between the control group and experimental group ( Z=1.44, P=0.151; Z=1.52, P=0.130). After treatment, there were no statistically significant differences in white blood cell count [4.28 (2.50)×10 9/L vs. 4.25 (1.88)×10 9/L] and neutrophil count [2.99 (2.50)×10 9/L vs. 2.94 (1.61)×10 9/L] between the two groups ( Z=0.67, P=0.503; Z=0.69, P=0.489). There were no statistically significant differences in white blood cell count and neutrophil count between the patients after treatment and before treatment in the two groups ( Z=0.77, P=0.443; Z=1.08, P=0.279; Z =1.06, P=0.289; Z=0.68, P=0.495). Before treatment, there were no statistically significant differences in serum inflammation indexes PCT [0.02 (0.03) μg/L vs. 0.02 (0.05) μg/L] and IL-6 [0.04 (0.21) μg/L vs. 0.04 (0.12) μg/L] between the two groups ( Z=0.70, P=0.482; Z=0.77, P=0.439). After treatment, there were statistically significant differences in serum inflammation indexes PCT [0.06 (0.17) μg/L vs. 0.03 (0.08) μg/L] and IL-6 [0.10 (0.25) μg/L vs. 0.01 (0.08) μg/L] between the two groups ( Z=2.08, P=0.038; Z=2.92, P=0.003). There were no statistically significant differences in serum inflammation indexes PCT and IL-6 in the experimental groups after treatment compared with before treatment ( Z=1.20, P=0.230; Z=1.19, P=0.235). In the control group, the serum inflammation index PCT level increased after treatment compared with before treatment, with a statistically significant difference ( Z=2.82, P=0.005), and the serum inflammation index IL-6 level increased compared with before treatment, but with no statistically significant difference ( Z=1.41, P=0.158). During the treatment, the incidence of radioactive esophagitis in the two groups was mainly grade Ⅰ-Ⅱ, with 24 cases in the control group and 28 cases in the experimental group. There were 8 patients with grade Ⅲ-Ⅳ radioactive esophagitis in the control group and 1 in the experimental group. There was a statistically significant difference in the occurrence of radioactive esophagitis between the two groups ( χ2=10.34, P=0.035). During the treatment period, most patients with radioactive pneumonia were rated as grade 0. There were 10 cases of mild radioactive pneumonia (grade Ⅰ-Ⅱ) in the control group had and 6 cases in the experimental group. There were 2 cases of grade Ⅲ radioactive pneumonia in the control group and experimental group respectively. There was no grade Ⅳ radioactive pneumonia in either group. There was no significant difference in radioactive pneumonia between the two groups ( χ2=1.34, P=0.720). Conclusion:Xiyanping injection can prevent the rise of serum inflammatory index PCT and reduce the severity of radioactive esophagitis in patients with esophageal cancer after treatment.

Objective:To observe any effect of intermittent theta burst stimulation (iTBS) on the spatially-delayed responses of working memory using cynomolgus macaques.Methods:The working memory of six male cynomolgus macaques (8-9 years old) was trained using a spatially-delayed response task. They were then randomly divided into an iTBS group and a control group, each of 3. The iTBS group was given iTBS at an intensity of 35% of the maximum output, with 2 seconds of stimulation followed by 8 seconds of rest with trains of 50Hz bursts repeated at a frequency of 5Hz over a period of 192 seconds once daily for 5 days, while the control group was given sham iTBS. Before and after the 5 days, the body weight and working memory of each animal were evaluated. The total number of effective feeding episodes, and of effective feeding episodes with short and long delay periods were recorded.Results:There was no significant change in the average body weight of either group before and after the modeling and iTBS intervention. After the intervention the number of total effective feeding cases and those with a short delay period were both significantly higher in the iTBS group than in the control group. However, no significant inter-group differences in the effective feeding cases with a long delay period were observed.Conclusions:iTBS is effective in improving the spatially-delayed responses of working memory, at least in cynomolgus macaques.

Objective:To understand the current situation and influencing factors of psychological stress reaction in patients with multi-drug resistance tuberculosis (MDR-TB) and to analyze the moderating effect of social support level between medical coping style and psychological stress reaction.Methods:This study was a cross-sectional study. Using the convenient sampling method, a total of 273 patients with MDR-TB who were hospitalized in the Tuberculosis Department of West China Hospital, Sichuan University from July 2019 to December 2020 were selected as the research objects. The Impact of Event Scale-Revised (IES-R) , Social Support Rating Scale (SSRS) and Medical Coping Modes Questionnaire (MCMQ) were used to investigate them. The χ 2 test and t test were used for univariate analysis, binomial logistic regression analysis was used for multivariate analysis and PROCESS V 3.3 software was used for moderating effect analysis. Results:A total of 37.36% (102/273) of MDR-TB patients had severe psychological stress reaction. Logistic regression analysis showed that age ( OR=1.039, 95% CI: 1.003-1.076) , marital status ( OR=0.136, 95% CI: 0.024-0.773) , occupation ( OR=0.057, 95% CI: 0.011-0.312) , daily alcohol consumption ( OR=28.968, 95% CI: 2.387-351.609) , social support level ( OR=0.927, 95% CI: 0.866-0.993) and patients' yielding psychology in the face of disease ( OR=2.216, 95% CI: 1.761-2.788) were independent factors affecting the psychological stress response of MDR-TB patients. The level of social support had significant moderating effects on avoidance (β=-0.132, P=0.002) , confrontation (β=-0.068, P=0.038) and psychological stress in MDR-TB patients. The more the patients with high social support level adopted the coping style of facing or avoiding, the lower the score of the event impact scale, that was, the weaker the psychological stress reaction. Conclusions:Medical staff should pay attention to the psychological stress reaction of MDR-TB patients and conduct targeted psychological counseling to enhance social support level of patients and improve treatment compliance, thereby reducing the recurrence rate of tuberculosis.