Background: Disrupted circadian rhythms have been associated with the development of irritable bowel syndrome (IBS). In some IBS patients, the symptoms may present with circadian fluctuations. Enterochromaffin cells (EC cells) and tryptophan hydroxylase 1 (TPH1) - 5 - hydroxytryptamine (5 - HT) signaling pathway are currently recognized as the key pathophysiological mechanism of IBS. Aims: To explore whether Bmal1, the core circadian clock gene, is involved in the occurrence of IBS by regulating TPH1-5-HT signaling pathway in EC cells. Methods: Normal Sprague-Dawley (SD) rats and IBS-model SD rats, as well as wild type (WT) and intestine-specific Bmal1 knockout (Bmal1

Background: Symptoms of irritable bowel syndrome with diarrhea (IBS-D) are known to be influenced by circadian oscillation; however, the pathophysiological mechanism is still unclear. Aims: To investigate the role and underlying mechanism of colon circadian clock gene Bmal1 involved in the occurrence of symptoms in IBS-D patients. Methods: Forty-six patients with IBS-D and 34 normal controls from Army Medical Center of PLA during September 2018 to February 2021 were recruited in this study. IBS-severity scoring system (IBS-SSS) was used to evaluate the severity of IBS-D symptoms. A colonoscopy was performed to obtain biopsy specimens from rectosigmoid colon. The concentration of 5-hydroxytryptamine (5-HT), and expressions of Bmal1 and chromogranin A (CgA), a biomarker of enterochromaffin cells (EC cells), in colonic mucosa were detected by ELISA and double-labeled immunofluorescence, respectively. Results: Both the 5-HT concentration and number of EC cells in colonic mucosa of IBS-D patients were significantly higher than those of the normal controls (all P< 0.05). Bmal1 was mainly expressed in intestinal epithelial cells and was highly expressed in EC cells. Co-expression of Bmal1 and CgA was observed. Compared with the normal control group biopsied at the same time point, expression of Bmal1 was significantly higher in specimens taken at 9 a.m., and expression of Bmal1 was significantly lower in specimens taken at 17 p.m. in IBS-D patients (all P< 0.05). Spearman correlation coefficient analysis showed that Bmal1 expression at 9 a.m. was positively correlated with the total score of IBS-SSS and subscore of abdominal pain and discomfort (r

Background: Visceral hypersensitivity is considered as a key pathophysiological mechanism involved in functional gastrointestinal disorders (FGIDs). Visceral nociception and hyperalgesia is existed extensively following exposure to post-traumatic stress disorder (PTSD), however, its molecular mechanism in intestinal tract is unclear. Aims: To explore the potential role of N-Myc downstream-regulated gene 2 (NDRG2) in intestinal tract for mediating visceral hypersensitivity following exposure to PTSD. Methods: PTSD model was established by single prolonged stress (SPS). SD rats were divided into normal control group, CTX group, PTSD group and PTSD+CTX group. Mice were divided into normal control group, PTSD group, NDRG2

Objective:To investigate the knowledge of the disease and demands of medical intervention in high-risk individuals of arteriosclerotic cardiovascular diseases (ASCVD).Methods:The 10-year ASCVD incidence risk prediction model was used to screen ASCVD high-risk individuals from Luohu district of Shenzhen city. From October 2018 to April 2019,a semi-structured in-depth interview was conducted among ASCVD high-risk individuals selected by stratified sampling method according to age, gender and educational level. The original data were analyzed with Colaizzi′s seven-step analysis method.Results:Total 37 interviewees were enrolled with an average age of (65.2±8.9) years and with an average ASCVD risk value of (14.2±3.2). Three themes were extracted from the interview, including: (1) Majority interviewees had better Knowledge about the hazards and risk factors of ASCVD; (2) Most of the interviewees had lower medical demands; (3) The interviewees were more likely to focus on symptomatic diseases or diseases disturbing them.Conclusions:The asymptomatic high-risk ASCVD individuals generally have better awareness of ASCVD and less demands for intervention. The result indicates that for health education, not only the knowledge, but also the attitude and behavior should be enhanced.

Objective: To study the effects of hedysarum polybotys saccharides (HPS) and selenizated hedysarum polybotys saccharides (SE-HPS) on the oral squamous cancer cell line SCC25. Methods: Different concentrations (0, 10, 25, 50, 100, 200, 400 μg/ml) of HPS and SE-HPS were added to SCC25 cells in the logarithmic growth stage. Cell proliferation was detected by the CCK-8 method, apoptosis was detected by flow cytometry, and apoptosis-related indexes were observed by RT-qPCR and Western blotting. Results : The concentrations of HPS and SE-HPS inhibited the proliferation of SCC25 cells. The inhibitory effect of 50 μg/mL HPS and SE-HPS on the proliferation of SCC25 cells was the strongest and was time-dependent. The inhibition effect significantly increased within 48 h, and the effect was achieved after 48 h. At the plateau stage, SE-HPS inhibited the proliferation of SCC25 cells more strongly than HPS (P < 0.05). The results of flow cytometry showed that 50 μg/mL HPS and SE-HPS acted on SCC25 cells for 48 h, and the apoptotic rates were 25.8% and 30.8% respectively. Compared with the control group (0 μg/mL HPS and SE-HPS), the difference was statistically significant (P < 0.05). RT-qPCR and Western blotting showed that 50 μg/mL HPS and SE-HPS acted on SCC25 cells for 48 h, and the mRNA and protein expression levels of the apoptosis gene Fas/FasL were upregulated. The difference was statistically significant (P < 0.05). Conclusion Both HPS and SE-HPS can inhibit the proliferation of SCC25 oral cancer cells, but SE-HPS is superior to HPS and can induce apoptosis through the Fas/Fasl pathway.

Objective: To investigate the correlation between serum lysosomal-associated membrane protein 2 (LAMP-2), LAMP-2 antibody levels and the progression of renal function in patients with chronic kidney disease (CKD). Methods: A total of 80 patients with CKD 3 to 5 stage (CKD group) and 50 healthy controls (control group) from August 2016 to August 2017 in Affiliated Dongfeng Hospital, Hubei University of Medicine were enrolled. The levels of hemoglobin, creatinine, urea, albumin, LAMP-2 and LAMP-2 antibody in fasting elbow venous blood of 2 groups were detected, and the estimate glomerular filtration rate (eGFR) was calculated. The CKD patients were followed up for 1 year, and renal function deterioration was defined as eGFR declined more than average value; the follow-up was over when the patients started dialysis or died, and those patients also defined as renal function deterioration. The patients were divided into high level group and low level group according to the serum LAMP-2 and LAMP-2 antibody levels. Results: The eGFR, hemoglobin and albumin in CKD group were significantly lower than those in control group: (24.60 ± 5.79) ml/min vs. (119.20 ± 9.52) ml/min, (111.36 ± 24.41) g/L vs. (144.60 ± 17.85) g/L and (36.83 ± 3.84) g/L vs. (45.92 ± 6.37) g/L, the creatinine, urea, LAMP-2 and LAMP-2 antibody were significantly higher than those in control group: (306.17 ± 49.24) μmol/L vs. (83.24 ± 5.55) μmol/L, (15.17 ± 3.39) mmol/L vs. (5.57 ± 1.33) mmol/L, (24.76 ± 5.47) μg/L vs. (12.93 ± 4.43) μg/L and (20.33 ± 4.89) μg/L vs. (9.98 ± 2.20) μg/L, and there were statistical differences (P<0.01). All 80 patients with CKD patients completed follow-up, and the follow-up time was 3 to 12 (8.14 ± 1.95) months. The eGFR at the end of followed-up was significantly lower than that at enrolment: (19.38 ± 7.30) ml/min vs. (24.60 ± 5.79) ml/min, the creatinine and LAMP-2 antibody at the end of followed-up were significantly higher than those at enrolment: (397.56 ± 52.32) μmol/L vs. (306.17 ± 49.24) μmol/L and (22.35 ± 4.74) μg/L vs. (20.33 ± 4.89) μg/L, and there were statistical differences (P<0.01); there were no statistical differences in hemoglobin, albumin, urea and LAMP-2 between the end of follow-up and enrollment (P>0.05). Among the 80 patients with CKD, renal function deterioration was in 42 cases, and renal function stability was in 38 cases. In renal function deterioration patients, the eGFR and urea at the end of followed up were significantly lower than those at enrolment: (18.28 ± 6.92) ml/min vs. (24.46 ± 5.76) ml/min and (13.51 ± 1.92) mmol/L vs. (14.81 ± 3.32) mmol/L, the creatinine, LAMP-2 and LAMP-2 antibody at the end of followed up were significantly higher than those at enrolment: (412.47 ± 53.21) μmol/L vs. (303.16 ± 47.87) μmol/L, (29.07 ± 5.42) μg/L vs. (25.89 ± 5.39) μg/L and (25.03 ± 3.30) μg/L vs. (20.95 ± 4.86) μg/L, and there were statistical differences (P<0.01 or <0.05); there were no statistical differences in hemoglobin and albumin between the end of follow-up and enrolment (P>0.05). In renal function stability patients, the eGFR at the end of follow-up was significantly lower than that at enrolment: (20.38 ± 7.58) ml/min vs. (24.73 ± 5.89) ml/min, the creatinine at the end of follow-up was significantly higher than that at enrolment: (381.07 ± 46.64) μmol/L vs. (309.49 ± 51.15) μmol/L, and there were statistical differences (P<0.01); there were no statistical differences in hemoglobin, albumin urea, LAMP-2 and LAMP-2 antibody between the end of follow-up and enrolment (P>0.05). In CKD patients, 38 cases were in LAMP-2 high level group (≥ 24.75 μg/L), and 42 cases were in LAMP-2 low level group (<24.75 μg/L); 38 cases were in LAMP-2 antibody high level group (≥ 20.33 μg/L), and 42 cases were in LAMP-2 antibody low level group (<20.33 μg/L). Kaplan-Meier curve analysis result showed that the renal function deterioration risk in LAMP-2 high level group was significantly higher than that in LAMP-2 low level group, LAMP-2 antibody high level group was significantly higher than that in LAMP-2 antibody low level group, and there were statistical differences (P<0.01). Conclusions Serum LAMP-2, LAMP-2 antibody levels are increased in patients with CKD, and higher serum LAMP-2 and LAMP-2 antibody levels may be associated with high risk of adverse kidney outcomes and become a promising marker to predict CKD progression.

Objective: To investigate the protective effect of intraperitoneal transplantation of human liver-derived stem cells at different times against concanavalin A (ConA)-induced acute liver injury in mice. Methods: A total of 88 male C57BL/6 mice were randomly divided into normal control group (group C), ConA model group (group M), and human liver-derived stem cells (HYX1)+ConA group (group E); according to the interval between phosphate buffer/HYX1 injection and ConA injection, Groups M and E were further divided into 3-hour groups (M1 and E1 groups), 6-hour groups (M2 and E2 groups), 12-hour groups (M3 and E3 groups), 24-hour groups (M4 and E4 groups), and 48-hour groups (M5 and E5 groups). The levels of alanine aminotransferase (ALT), aspartate transaminase (AST), and total bilirubin (TBil) in peripheral blood were measured, liver tissue sections were used to observe pathological changes, and the Ishak score for liver inflammation was determined. The independent samples t-test was used for comparison between groups, and P < 0.05 was considered statistically significant. Results: The levels of ALT, AST, and TBil in group C were (36.25±1.16) U/L, (120.20±5.77) U/L, and (2.20±0.23) μmol/L, respectively; the levels of ALT, AST, and TBil and Ishak score were (8 721.23±837.39) U/L, (8 110.31±290.10) U/L, (8.41±0.10) μmol/L, and (13.32±1.30), respectively, in group M1, (8 334.31±666.50) U/L, (7 560.20±760.34) U/L, (10.40±0.80) μmol/L, and (12.67±0.81), respectively, in group M2, (8 960.75±551.93) U/L, (8 535.62±675.14) U/L, (10.95±1.43) μmol/L, and (14.57±0.65), respectively, in group M3, (8 618.57±886.40) U/L, (11 440.54 ± 1 327.86) U/L, (13.30±1.86) μmol/L, and (13.21±1.06), respectively, in group M4, and (10 170.13±1 112.37) U/L, (11 470.56±1 108.40) U/L, (12.75±1.55) μmol/L, and (15.07±1.58), respectively, in group M5. The levels of ALT, AST, and TBil and Ishak score were (1 016.35±163.47) U/L, (952.30±103.91) U/L, (7.77±0.62) μmol/L, and (3.50±0.21), respectively, in group E1, (42.10±6.20) U/L, (126.72±13.33) U/L, (3.41±0.53) μmol/L, and (2.01±0.40), respectively, in group E2, (44.21±4.30) U/L, (216.71±35.88) U/L, (3.47±0.44) μmol/L, and (2.13±0.25), respectively, in group E3, (2 909.69±212.14) U/L, (2 988.43±333.70) U/L, (7.03±0.93) μmol/L, and (4.70±0.50), respectively, in group E4, and (7 874.26±799.60) U/L, (10 940.54±947.35) U/L, (10.53±1.09) μmol/L, and (8.60±0.83), respectively, in group E5. Groups M1-M5 had significantly higher levels of ALT, AST, and TBil than group C (all P < 0.01), and groups M1-M4 had significantly higher levels of AST and ALT than groups E1-E4 (all P < 0.01), while there were no significant differences in the levels of AST and ALT between groups M5 and E5 (both P > 0.05). The pathological sections of liver tissue showed that compared with group M, group E had significant reductions in the degree of necrosis and Ishak score (both P < 0.05). Conclusion Intraperitoneal transplantation of human liver-derived stem cells has a protective effect against ConA-induced acute liver injury in mice, and the injection at 6 and 12 hours in advance has the best protective effect.

Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Cell Line, Tumor ; drug effects ; Humans ; Leukemia ; pathology ; Oxides ; pharmacology

OBJECTIVETo study the blocking effect of CXCR4 inhibitor AMD3100 on the adhesion of leukemia cells to osteoblast niche, and the reversal of multidrug resistance in leukemia cells.

METHODSMesenchymal stem cells (MSCs) from leukemia patients were planted on the bio-derived bone scaffolds and then induced into osteoblasts to establish the bio-osteoblast niche. The levels of SDF-1were tested with ELISA. The leukemia cell line MV4-11 cells with FLT3-ITD mutation were inoculated into the bio-osteoblast niche to build a three-dimensional co- culture system. The expression level of CXCR4, adhesion and apoptosis rates of leukemia cells were observed by flow cytometry after incubation with AMD3100 and Ara-C for 24 h and 48 h.

RESULTS(1)The supernatant levels of SDF-1 in cultured osteoblast were (304 ± 18), (410 ± 28) and (396 ± 16) pg/ml on 7 th, 14 th and 21 th day, respectively. It reached the highest on 14 th day. The expression level of CXCR4 in cultured MV4-11 cells was (72 ± 16)%. (2)Adhesion rate of MV4-11 cells to osteoblast niche was (40.1 ± 8.1)% after AMD3100 treatment for 24 h, while that of control group was (65.6 ± 12.1)% (P<0.05). (3)The apoptosis rate of MV4-11 cells incubated with AMD3100 for 24 h was (5.6 ± 0.8)%, while that of control group was (2.5 ± 0.5)%. The apoptosis rates of AMD3100-induced MV4-11 cells were (10.0 ± 2.4)%, (17.8 ± 2.3)% and (25.1 ± 2.4)% after treatment with Ara-C at 0.02, 0.20, 2.00 mg/ml respectively and they were (6.7 ± 1.0)%, (10.3 ± 1.5)%, (16.2 ± 3.1)% respectively in AMD3100-noninduced control group, the difference was significant (P<0.05).

CONCLUSIONAMD3100 can block the interaction between osteoblasts niches and leukemia cells, and play an important role in the reversal of multidrug resistance in leukemia cells.

Apoptosis ; Cell Adhesion ; Cell Line, Tumor ; Chemokine CXCL12 ; Coculture Techniques ; Cytarabine ; Drug Resistance ; Flow Cytometry ; Heterocyclic Compounds ; Humans ; Leukemia ; Mesenchymal Stromal Cells ; Osteoblasts ; Receptors, CXCR4 ; Signal Transduction