Synthesis, Detection, and Metabolism of Pyridone Ribosides, Products of NAD Overoxidation.

Pyridone-containing adenine dinucleotides, ox-NAD, are formed by overoxidation of nicotinamide adenine dinucleotide (NAD+) and exist in three distinct isomeric forms. Like the canonical nucleosides, the corresponding pyridone-containing nucleosides (PYR) are chemically stable, biochemically versatile, and easily converted to nucleotides, di- and triphosphates, and dinucleotides. The 4-PYR isomer is often reported with its abundance increasing with the progression of metabolic diseases, age, cancer, and oxidative stress. Yet, the pyridone-derived nucleotides are largely under-represented in the literature. Here, we report the efficient synthesis of the series of ox-NAD and pyridone nucleotides and measure the abundance of ox-NAD in biological specimens using liquid chromatography coupled with mass spectrometry (LC-MS). Overall, we demonstrate that all three forms of PYR and ox-NAD are found in biospecimens at concentrations ranging from nanomolar to midmicromolar and that their presence affects the measurements of NAD(H) concentrations when standard biochemical redox-based assays are applied. Furthermore, we used liver extracts and 1H NMR spectrometry to demonstrate that each ox-NAD isomer can be metabolized to its respective PYR isomer. Together, these results suggest a need for a better understanding of ox-NAD in the context of human physiology since these species are endogenous mimics of NAD+, the key redox cofactor in metabolism and bioenergetics maintenance.

Frailty in chronic myeloid leukemia: evidence from 2016-2018 Nationwide Inpatient Sample of the US.

BACKGROUND: Frailty is a marker of poor prognosis in older adults with hematologic malignancies and contributes to the severe vulnerability of the aging population to adverse health outcomes. This study aimed to determine the association between frailty and outcomes in hospitalized patients with chronic myeloid leukemia (CML). METHODS: The International Classification of Diseases (ICD-10) identified data on hospitalized patients 20 years or older admitted with CML between 2016 and 2018 in the US National Inpatient Sample (NIS) database. The cohort was further divided into groups of patients with or without frailty. Logistic regression analysis was performed to determine associations between study variables and clinical outcomes. A stratified analysis of the association between frailty and in-hospital mortality by age group was also performed. RESULTS: A total of 13,849 hospitalized patients with CML were included, 49.6% of whom had frailty. The mean age of the patients was 65.1 years, and 7,619 (56.2%) of them were male. Frailty was associated with nearly 4 times the risk of in-hospital mortality, 3 times the risk of unfavorable discharge, 3 times the risk of prolonged LOS,, and significantly more in total hospital costs. In addition, frailty was associated with a significantly increased risk of in-hospital mortality in all age subgroups (< 40 years, 40-59 years, and > 60 years) compared with no frailty. CONCLUSIONS: Frailty strongly predicts poor clinical outcomes in US patients with CML.

Abnormalities in acute salivary biochemical characteristic responses to gustatory stimulation with citric acid in chronic non-atrophic gastritis.

BACKGROUND AND AIM: Salivary characteristics are altered in gastrointestinal diseases and related to oral taste disorder. However, specific salivary biochemical characteristics and their relationships with oral taste disturbances in chronic non-atrophy gastritis (CNAG) remain uncertain. METHODS: Seventy patients with CNAG and 70 subjects in healthy control group (HCG) were enrolled in our study. The levels of salivary flow rate (SFR), pH, salivary α-amylase (sAA) activity, total protein density (TPD), chloride concentration, and calcium concentration were determined before and after citric acid stimulation and compared between CNAG with and without oral taste disturbances. RESULTS: Average body mass index (BMI) of CNAG (17.75 ± 2.08) was lower than that of HCG (21.96 ± 1.72, P < 0.01). Compared with HCG, CNAG showed increased TPD and calcium concentration but decreased SFR both before and after acid stimulation (P < 0.01), as well as reduced sAA and salivary chloride responses to acid stimulation (P < 0.01). Compared with CNAG with normal BMI (24.29%, 17/70), sAA activity response to acid stimulation was reduced in those with low BMI (75.71%, 53/70, P < 0.05). Under resting condition, CNAG with dry mouth (55.71%, 39/70) showed increased SFR and decreased TPD (P < 0.05), as compared with CNAG without dry mouth (44.29%, 31/70). Compared with CNAG without bitter taste (57.14%, 40/70), pH was decreased in those with bitter taste (42.86%, 30/70) under both resting and stimulated conditions (P < 0.05). CONCLUSION: Decreased sAA activity may reflect malnutrition state and be one potential marker of poor digestion, decreased salivary pH may contribute to bitter taste perception, and reduced TPD might be a cause of dry mouth in CNAG.

[Effects of Methanol Extracts from Atractylodes macrocephalae Rhizoma on Small Intestinal Epi- thelial Cell Proliferation and Migration, and Expression of Phospholipase C-γl].

OBJECTIVE: To observe the effects of methanol extracts from Atractylodes macro- cephalae Rhizoma (AMR) on the proliferation and migration of IEC-6 cell (small intestinal epithelial cells) and the expression of phospholipase C-γ1 (PLC-γ1) , and to explore the mechanism of AMR (a Chinese herb capable of invigorating Pi replenishing qi) for promoting repair of gastrointestinal mucosal injury. METHODS: IEC-6 cells were divided into the blank group, the positive control (spermidine, SPD; 5 µmol/L) group, AMR extracts groups (50, 100, and 200 mg/L). The alpha-difluoromethylornithine (DFMO, polyamines synthesis inhibitor) group, the SPD +DFMO group, AMR extracts (50, 100, and 200 mg/L) +DF- MO groups were set up in stress test. IEC-6 cells were cultured by adherence for 24 h,and then treated with AMR extracts for appropriate periods of time. Effects of IEC-6 cell proliferation after action of AMR extracts were detected by Real-time Cell Analyzer (RTCA). The effect of AMR extracts on IEC-6 cell migration number was detected using scratch method. mRNA and protein expressions of PLC-γ1 levels were detected by fluorescent quantitative polymerase chain reaction ( RT-qPCR) and Western blot respectively. RESULTS: Compared with the blank group, AMR extracts showed no obvious effect on IEC-6 cell proliferation (P >0. 05). But SPD and AMR extracts (100 and 200 mg/L) not only promoted IEC-6 cell migration (P <0. 01), but also improved mRNA and protein expressions of PLC-γl in the process of cell migration (P <0. 01). Compared with the DFMO group, SPD and AMR extracts (100 and 200 mg/L) could reverse inhibitory effects of DFMO on cell migration, and mRNA and protein expressions of PLC-γl (all P <0. 01). CONCLUSION: AMR extracts played roles in repairing gastrointestinal mucosal injury possibly by promoting polyamine mediated intestinal epithelial cell migration, and its effect on intestinal epithelial cell proliferation was not main potentcy.

[Effect of Sijunzi Decoction Polysaccharide on IEC-6 Cell Migration,Potassium Channel and Membrane Potential].

Objective: To investigate the effect of Sijunzi decoction polysaccharide( SJZDP) on intestinal epithelial cells( IEC-6)cell migration and polyamine signaling pathway potassium channel during intestinal epithelial cell migration, and to explore the mechanism of SJZDP on promoting gastrointestinal mucosal restitution after wounding. Methods: Cell migration model was established by scratch damage, and then the effect of SJZDP normal cultured or with difluoromethylornithine( DFMO) and 4-aminopyridine( 4-AP) on IEC-6 cell migration was observed and calculated on this wounding model. The effect of SJZDP on expression of IEC-6 cell kv1. 1 mRNA and protein levels were detected by RT-q PCR and Western blot analysis, respectively. The Effects of SJZDP on IEC-6 cell membrane potential were detected by flow cytometry. Results: The results showed that treatment with SJZDP( 40,80,160 mg / L) caused a promotion of IEC-6 cell migration,and increased of expression of in IEC-6 cell kv1. 1 mRNA and protein significantly( P < 0. 05 or P < 0. 01) compared with normal control group. In addition, SJZDP( 40,80,160 mg / L) increased cell membrane potential which resulted in cell membrane hyperpolarization compared with normal control group( P < 0. 05 or P < 0. 01). SJZDP( 40,80,160 mg / L) reversed the inhibition of cell migration was reduced,kv1. 1 mRNA,kv1. 1 protein expression, and cell membrane potential were decreased by polyamines synthesis inhibitor DFMO compared with DFMO model group( P < 0. 05 or P < 0. 01). SJZDP( 20,40,80 mg / L) reversed the inhibition of cell migration,kv1. 1 protein and mRNA levels expression were decreased by potassium channel inhibitor 4-AP compared with 4-AP model group( P < 0. 05 or P < 0. 01). Conclusion: These results indicate that the effect of SJZDP on promoting IEC-6 cell migration may be related to its influence on polyamine signaling pathway potassium channel and cell membrane potential.

[Comparative Study on Effects of Anti-Gastritis,Gastric Mucosal Protection and Gastrointestinal Movement Promotion of Root and Stem of Zanthoxylum nitidum].

Objective: To provide the experimental evidence for expansion of medicinal parts of Zanthoxylum nitidum by comparing the effects of anti-gastritis,gastric mucosal protection and gastrointestinal movement promotion of its root and stem. Methods: The pharmacological effects between root and stem of Zanthoxylum nitidum were compared by observing the anti-gastritis effect on rats with chronic superficial gastritis induced by iodoacetamide, evaluating the gastric mucosal protective effect on rats' gastric ulcer induced by stress, indometacin and pylorus ligation test, and investigating gastrointestinal movement promotion effect on mice gastric evacuation and intestinal propelling. Results: Both root and stem of Zanthoxylum nitidum showed effects of relieving the inflammation symptoms of rats' gastric mucosa induced by iodoacetamide, gastric ulcer respectively induced by stress, and presenting a strong inhibition of free acid and pepsin activity in gastric juice. Furthermore stem parts of Zanthoxylum nitidum in promoting gastrointestinal motility even showed better efficacy than root. Conclusion: Stem of Zanthoxylum nitidum has similar effects of anti-gastritis, gastric mucosal protection and gastrointestinal movement promotion with root of Zanthoxylum nitidum.

A highly sensitive electrochemiluminescence immunoassay for detecting human embryonic human chorionic gonadotropin in spent embryo culture media during IVF-ET cycle.

PURPOSE: To investigate the stability and repeatability of electrochemiluminescence immunoassay (ECLIA) for beta-hCG detection in embryo spent culture media. To evaluate the correlation between the viability of preimplantation embryo and beta-hCG profile by the new assay. METHODS: In a retrospective study, a total of 357 spent culture media from day1 to day5 were individually collected and quantified by ECLIA. The blank controls and reliability test were performed with normal saline/pure culture media. RESULTS: 1) There was no detectable amount of beta-hCG in blank controls. A high degree of linearity (R(2) = 0.995) was found in this study; intra-assay and inter-assay coefficient of variation were 4.87 % and 6.25 %. 2) A significantly higher concentration of beta-hCG was found at day5 group than it at day3 group, both in total samples (1.47 ± 0.68mIU/ml vs 0.55 ± 0.32mIU/ml) and in homologous embryo samples (1.43 ± 0.91mIU/ml vs 0.52 ± 0.23mIU/ml). 3) There was a positive correlation between beta-hCG concentration and implantation rate (r = 0.559 at day3 and 0.535 at day5) or blastocyst morphological grading (r = 0.411). CONCLUSIONS: ECLIA may be an optimal choice for detecting beta-hCG in spent culture media to assess embryo viability, indicating secreted beta-hCG as a useful biomarker for embryo selection in IVF-ET procedure.

[Effect of sijunzi decoction on salivary amylase secretion disorder and VIP-cAMP signaling pathway in splenasthenic rats].

OBJECTIVE: To observe the effect of Sijunzi Decoction on secretion disorder of salivary amylase in splenasthenic rat and its mechanism. METHODS: The model group rats received reserpine 0.5 mg/kg through subcutaneous injection while the control group rats received the same volume of saline for 8 days. After being modeled, the model group were divided into treatment group and model control group, treatment group were given orally Sijunzi Decoction, model control group and normal group were fed the same amount of distilled water for 4 weeks. The animal were anaesthetized and the left parotid was removed, the wounds were sutured. When the animals were awake but drowsy, 20 microL 10% glacial acetic acid was applied on the apex of the tongue once a minute for 30 minutes, removed the right parotid gland of the animals. The samples were frozen and amylase activity and VIP, cyclic adenosine monophosphate (cAMP) content and VAMP-8, SNAP-23 protein expression in the parotid glands were detected. RESULTS: Change of sAA in parotid acinar was not significantly different between treatment group and normal groups, but higher in model control groups after acid stimulation. The VIP and PKA contents were not significantly different among three groups. VIP, cAMP content and PKA activity increased significantly in normal group while VIP increased slightly, cAMP and PKA activity decreased in model control groups, which returned to some degrees in treatment group after acid stimulation. Expression of VAMP-8 protein was not significantly different between treatment group and model control groups, while expression of SNAP-23 was lower in model control groups, expression of VAMP-8 and SNAP-23 was higher in treatment group than which in model control groups. CONCLUSION: Sijunzi Decoction has a certain effect on secretion disorder of salivary amylase in splenasthenic rat, which mechanism may be related to recover changes of VIP-cAMP signal pathway in the splenasthenic rat's parotid gland cells,including increase VIP content and expression of VAMP-8 and SNAP-23.

[Effect of polysaccharides from Radix Glycyrrhizae on migration and polyamines contents of IEC-6 cell].

OBJECTIVE: To study the effect of polysaccharides from Radix Glycyrrhizae on migration and polyamines (putrescine, spermidine and spermine) contents of IEC-6 cell. METHODS: Cell migration model was induced by scratch method in each well,and the polyamines in IEC-6 cell was determined by pre-column derivation high performance liquid chromatography. The polysaccharides inhibited effect on migration and polyamines contents of IEC-6 cells, and on IEC-6 cell migration by DFMO (a polyamines synthesis inhibitor) and the polyamines contents in the cells were observed. RESULTS: The polysaccharides (50 mg/L or 100 mg/L) was able to promote the cell migration, reverse the cell migration inhibition by DFMO, enhance the IEC-6 cell polyamines (putrescine, spermidine and spermine) contents in the process of cell migration and reverse the reduction of polyamines (putrescine, spermidine and spermine) induced by DFMO. CONCLUSION: The effect of Radix Glycyrrhizae on the gastrointestinal mucosal damage repairing may be related to increasing polyamine content in cells and promoting cell migration.

Association between serum vitamin D levels and Helicobacter pylori cytotoxic-associated gene A seropositivity: a cross-sectional study in US adults from NHANES III.

OBJECTIVE: To assess the association of serum vitamin D (VD) levels and Helicobacter pylori (H. pylori) cytotoxic-associated gene A (CagA) seropositivity, and further explore potential effect modifiers in this association. DESIGN: Cross-sectional study. SETTING: Data from phase I of the National Health and Nutrition Examination Survey (NHANES III, 1988-1991) led by the Center for Disease Control and Prevention. PARTICIPANTS: A total of 3512 US adults (≥20 years) with both serum VD levels and H. pylori CagA antibody data from NHANES III were included in the analysis. METHODS: VD deficiency was defined as serum 25(OH)D concentrations<20 ng/mL. Logistic regression models were used to assess the association of serum VD levels and H. pylori CagA seropositivity (VD-Hp CagA+), and stratification analyses were used to explore potential effect modifiers. RESULTS: There was no significant association of VD-Hp CagA+ in the general population. But serum 25(OH)D concentrations were associated with H. pylori CagA+ in non-Hispanic whites (adjusted OR=1.02, 95% CI: 1.00 to 1.03), other races/ethnicities (adjusted OR=1.08, 95% CI: 1.01 to 1.06), populations born in other countries (adjusted OR=1.09, 95% CI: 1.04 to 1.15) or occasional drinkers (adjusted OR=0.93, 95% CI: 0.88 to 0.99). VD deficiency was associated with H. pylori CagA+ in non-Hispanic whites (adjusted OR=0.69, 95% CI: 0.53 to 0.92), populations born in other countries (adjusted OR=0.47, 95% CI: 0.25 to 0.89), non-drinkers (adjusted OR=0.80, 95% CI: 0.65 to 0.99), occasional drinkers (adjusted OR=2.53, 95% CI: 1.06 to 6.05), population with first quartile level of serum ferritin (adjusted OR=0.70, 95% CI: 0.51 to 0.96) or fourth quartile level of serum folate (adjusted OR=0.63, 95% CI: 0.46 to 0.87). CONCLUSIONS: Racial/ethnic differences and different serum ferritin or serum folate levels may be effect modifiers for the association of VD-Hp CagA+.

Implementation of the treat-to-target approach and treatment satisfaction in patients with rheumatoid arthritis: perspectives of Chinese rheumatologists.

OBJECTIVE: To investigate the implementation of treat-to-target (T2T) and treatment satisfaction from Chinese rheumatologists' perspectives. METHODS: This retrospective analysis of a cross-sectional database collected from rheumatologists and their adult patients with RA in China using Adelphi Real World Disease Specific Programme™ methodology. Multivariate logistic regression models were used to evaluate factors associated with T2T use, achievement of T2T goals, and physician treatment satisfaction. RESULTS: Sixty physicians provided data for 600 patients, of whom 39.0% (234/600) were being treated using T2T, and 64.9% (366/564) had achieved their T2T goal. Physicians were satisfied with treatment in 74.3% (445/599) of patients. Patients with a higher pain score were more likely to be managed using T2T (odds ratio (OR) 1.25; p = 0.017), but less likely to have achieved the T2T goal (OR 0.76; p = 0.004). T2T use was more likely if patients had a longer time since diagnosis (> 2 vs ≤ 2 years; OR 1.61; p = 0.031) or received targeted synthetic or biologic disease-modifying antirheumatic drugs (tsDMARDs or bDMARDs; OR 6.90; p < 0.001). T2T goal achievement was more likely for patients with higher body mass index (≥ 24 vs < 24 kg/m2; OR 2.73; p = 0.001) or full-time employment (OR 2.11; p = 0.005). Physician treatment satisfaction was more likely if the T2T goal was achieved (OR 4.78; p < 0.001) or tsDMARDs or bDMARDs were used (OR 2.58; p = 0.017), and less likely if pain scores were higher (OR 0.79; p = 0.019). CONCLUSIONS: T2T implementation in China is suboptimal. Our findings provide insight into T2T implementation and physician treatment satisfaction, supporting T2T use in Chinese RA clinical practice. Key Points • T2T implementation in China is currently suboptimal. • Patients with greater pain were more likely to be managed using T2T but were less likely to have achieved their T2T goals. • Physician treatment satisfaction was associated with T2T goal achievement.

Shugan Jieyu capsule improve sleep and emotional disorder in coronavirus disease 2019 convalescence patients: a randomized, double-blind, placebo-controlled trial.

OBJECTIVE: To evaluate the efficacy of the Shugan Jieyu capsule on improving sleep and emotional disorder during Coronavirus disease 2019 (COVID-19) convalescence. METHODS: We conducted a randomized, double-blind, placebo-controlled trial, and recruit 200 COVID-19 convalescence patients and then divide the subjects into two groups respectively: the experimental group ( 100) and the control group ( 100). Patients in the control group were given doses as a placebo, while those in the experimental group were given Shugan Jieyu capsule. The investigators mainly observed the differences between the two groups before and after treatment in terms of the rate of reduction and the rate of efficiency in Hamilton Depression Scale (HAMD-17) total scores from baseline, and recorded the scores of Hamilton Anxiety Scale (HAMA), Patient Health Questionnaire-15 (PHQ-15), Insomnia Severity Index (ISI) and Treatment Emergent Symptom Scale at 2 week, the 4 week and the 6 week respectively after treatment, and compared the differences between the groups. And the occurrence of adverse events was recorded. RESULTS: After 6-week treatment, there were statistically significant differences in the rate of reduction as well as efficiency in HAMD-17 scores, HAMA Total Scores, PHQ-15 Score, ISI Score from baseline in the experimental group and control group (< 0.05). There were 4 adverse events in the experimental group and 1 in the control group. CONCLUSION: Shugan Jieyu capsule could significantly improve sleep and emotional disorder in patients during COVID-19 convalescence.

The Single-Nucleotide Polymorphism of miR-27a rs895819 and the Expression of miR-27a in Helicobacter pylori-Related Diseases and the Correlation with the Traditional Chinese Medicine Syndrome.

Aims: The study aims to explore the effects of the single-nucleotide polymorphism of miR-27a and its expression in Helicobacter pylori (H. pylori)-related diseases and the relationship between gastric pathology and traditional Chinese medicine (TCM). Methods: Subjects were classified into six histopathological groups and five TCM syndrome groups. All specimens underwent H. pylori detection through rapid urease test and methylene blue staining. Histopathological characteristics were observed by hematoxylin-eosin. The expression of miR-27a and its genotype were, respectively, detected by Quantitative Real-Time PCR and direct sequencing. Results: H. pylori promoted the malignant evolution of gastric mucosa and were involved in the formation of TCM syndrome. In H. pylori-positive patients, the frequency of miR-27a CT genotype at the rs895819 locus and its expression in the gastric cancer group were higher than those in other pathological groups. TCM syndrome had a close relationship with histopathological changes, and patients with spleen-qi deficiency syndrome had a higher risk of gastric cancer than other syndromes, regardless of H. pylori infection. Conclusion: The C allele at miR-27a rs895819 locus may be an oncogene in gastric cancer. High levels of miR-27a could play an important role in gastric malignant evolution, especially cancerization. There is a certain connection between TCM syndrome and pathological changes of the gastric mucosa to some extent, where patients with SQD syndrome had a higher risk of GC.

Decreased salivary α-amylase activity responding to citric acid stimulation in Myasthenia gravis with malnutrition.

OBJECTIVES: Malnutrition, defined according to Nutritional risk screening (NRS 2002), is commonly observed in patients of Myasthenia gravis (MG), a neuromuscular disorder manifested by varied degrees of skeletal muscle weakness. Because biochemical composition of saliva changes in correspondence to alterations in nutritional status, we tested our hypothesis that a certain saliva component(s) might serve as a biomarker(s) for nutrition status of MG, particularly for those MG patients with high risk of malnutrition. MATERIALS AND METHODS: 60 MG patients and 60 subjects belonging to the healthy control group (HCG) were enrolled in this case-control study. The salivary α-amylase (sAA) activity, salivary flow rate (SFR), pH, total protein density (TPD), and the concentrations of chloride and calcium ions in MG group with or without malnutrition were measured before and after citric acid stimulation. Thereafter, the relationship between sAA activity and BMI was determined in MG and HCG. RESULTS: Compared with HCG, more patients with malnutrition, increased TPD and chloride and calcium concentrations but decreased pH value and SFR both before and after acid stimulation, as well as reduced sAA activity, pH and TPD responses to acid stimulation. MG with malnutrition showed decreased sAA activity and TPD responding to acid stimulation compared with those without malnutrition. Compared with normal BMI, sAA activity response to acid stimulation was reduced in low BMI. There was a significant strong positive correlation between the ratio of sAA activity and BMI in MG. CONCLUSIONS: Salivary biochemical characteristics are abnormally altered in MG with malnutrition. Altered sAA activity responding to acid stimulation was associated with malnutrition. CLINICAL RELEVANCE: Decreased sAA activity responding to acid stimulation can reflect malnutrition state and may be one potential screening marker for MG patients with high risk of malnutrition.

[Study on ribosomal protein gene expression in patients with ulcerative colitis of Pi-asthenic syndrome and Pi-asthenic syndrome].

OBJECTIVE: To study differential expression profiles of ribosomal protein (RP) genes in healthy subjects and ulcerative colitis (UC) patients of Pi-asthenic syndrome (PAS) and of dampness-heat syndrome (DHS), thus providing experimental bases for " Pi as the source of qi and blood" theory from the view of protein synthesis. METHODS: RP genes arrays were made. The mucous membrane of colon was detected in four UC patients of PAS (UC-PAS), four UC patients of DHS (UC-DHS), and four healthy subjects (N), and data analyzed using BRB-TOOL Software Package (3.9). Bioinformatics analyses were conducted in differential genes. RESULTS: Low-density RP gene chips were successfully produced, including 77 RP genes and two RP like genes (RPL26-like1 and RPL7-like1). There were twelve differential genes between UC (PAS+DHS) and N, all of which were down-regulated genes. There were nineteen differential genes between UC-DHS and N, all of which showed down-regulating tendency. There were three differential genes between UC-PAS and N, all of which were down-regulated genes. There were six differential genes between UC-PAS and UC-DHS, all of which were up-regulated genes. Cluster analysis showed that normal and UC samples of this chip can be classified according to gene expression profiles, and UC-PAS and UC-DHS can be classified by clustering. Various differential genes had a common transcription regulatory factor. CONCLUSIONS: RP genes arrays were successfully produced. RP gene expressions were down-regulated in UC-PAS and UC-DHS. Corresponding gene expression profiles were shown in N, UC-PAS and UC-DHS.

Studies on cell migration model in intestinal epithelial restitution for pharmacological research.

OBJECTIVE: To set up a suitable IEC-6 migration model for pharmacological research and observe the effect of complex polysaccharide from Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao to IEC-6 cell migration. METHODS: The main conditions related to the establishment of the model, including the planting density of the cell, the observation time after scratching, the concentration of the auxiliary material Matrigel, the treatment of the serum-starvation, the concentration of alpha-difluoromethylornithine (DFMO), an inhibitor of the cell migration, were investigated respectively; and the effects of the tested medicines on the model were observed. RESULTS: 4 x 10(5) cell/mL was the suitable planting density of the cell in the 6-well plate; at the 24th hour after scratching was the appropriate time to count the migrating cells; and the proper concentration of Matrigel was 5%; the serum-starvation could evidently reduce the migrating cells, so the culture medium should contain the serum; 2.5 - 5 mmol/L DFMO was proper for inhibition of the cell migration. Complex polysaccharide from Astragalus membranaceus and spermidine both can promote cell migration. CONCLUSION: The established model of IEC-6 cell migration was suitable for intestinal epithelial restitution such as the researches on pathophysiological mechanisms is the effects of the medicines on the cell migration.

Pharmacological treatments of Chinese herbal medicine for irritable bowel syndrome in adults: A network meta-analysis of randomized controlled trials.

INTRODUCTION: Plenty of clinical studies have suggested the value of Chinese herbal medicine (CHM) for patients with irritable bowel syndrome (IBS), but their efficacy and safety have not been systematically concluded yet. This article aimed to compare and rank the therapeutic effect and safety of CHM with routine pharmacotherapies and placebo in the treatment of IBS. METHODS: Randomized controlled trials regarding CHM to treat IBS were searched in six databases from inception to Jan 31, 2020. A network meta-analysis was conducted to analyze the data of included publications. The quality assessment was assessed by Cochrane Handbook and GRADEpro software. The risk ratio was calculated for dichotomous outcomes while the standardized mean difference was used for continuous variables with 95% credible intervals. A Funnel plot was performed to evaluate publication bias. The surface under the cumulative ranking curve was conducted to rank the included interventions. Data were analyzed with STATA 15.0 and Review Manager 5.3. RESULT: 3194 records were searched, and 28 eligible trials involving 3323 patients ere identified. Compared with conventional therapies and placebo, Jianpi-Chushi therapy showed significant improvement in adequate relief and IBS symptom severity scale; Shugan-Jianpi therapy showed the best efficacy in relieving the abdominal pain and abdominal distension; Wenshen-Jianpi therapy had a better effect on avoiding adverse effects and improving stool character. CONCLUSION: This study confirmed that CHM could be beneficial for patients with IBS in relieving their clinical symptoms and should be recommended as alternative therapies. The quality of evidence in this study based on the GRADE system was "low".

Efficacy and safety of Qingre-Chushi therapies in active ulcerative colitis: A network meta-analysis.

BACKGROUND: Ulcerative colitis (UC) is a chronic inflammatory disease with an increasing incidence in the world. Qingre-Chushi therapies (QC) can alleviate clinical symptoms. Therefore, a network meta-analysis was conducted to systematically evaluate the efficacy and safety of QC in the treatment of active UC patients. METHODS: 7 databases were screened and relevant randomized controlled trials were selected. The tools of Cochrane Handbook and the GRADE system were conducted to assess the quality of outcomes. Pooled risk ratio or standard mean difference was calculated with 95% credible interval for outcomes measurement using the random-effects model. The surface under the cumulative ranking curve (SUCRA) was performed to rank the treatments. The larger SUCRA scores, the more effective interventions. RESULTS: A total of 3560 articles were identified and 21 studies including 1829 participants were included for further analysis. Totally, 9 therapies regimens were compared: oral mesalazine, mesalazine enema, mesalazine suppository, oral mesalazine + mesalazine enema, oral QC, oral QC + oral mesalazine, QC enema, oral QC + QC enema, and oral mesalazine + QC enema. Based on the SUCRA plot, oral QC + oral mesalazine was the best treatment in inducing clinical response; oral QC + QC enema had the best efficacy in the improvement of Mayo scores and alleviating abdominal pain; oral mesalazine + mesalazine enema was the optimal therapy in the endoscopic improvement and reducing diarrhea; QC enema + oral mesalazine was the best option in preventing bloody stool. CONCLUSION: This study confirmed the efficacy and safety of QC in treating active UC and suggested that the combination of oral medications with topical can achieve more benefits.

[Effect and mechanism of reserpine for changing salivary protein secretion in Pi-deficient rats].

OBJECTIVE: To study the effect of reserpine (RSP) for changing salivary protein secretion in Pi-deficient rats and to explore its possible mechanism. METHODS: Twenty rats allocated in the RSP group were given subcutaneous injection of RSP [0.4 mg/(kg x d)] for 9 successive days, while the other 20 rats in the control group were injected with same volume of saline instead. On the 10th day, ten rats randomly selected from each group were subjected for extracting saliva to detect salivary amylase activity (sAA) before and after an acid stimulation; and drawing blood from the orbital vein to measure the contents of vasoactive intestinal peptide (VIP) and cyclic adenosine monophosphate (cAMP). Then they were sacrificed and their parotids were taken out for pathological examination with HE staining, as well as for VIP and cAMP measuring, and zymogen granules counting under a transmission electron microscope. The remainder animals were stopped injecting and normally fed to 40 days, then subjected to be detected as above-mentioned. RESULTS: Food intake and body weight reduction were more significantly in the RSP group than in the control group. On the 10th day, the ratio of sAA before/after stimulation in the RSP group was 0.39 +/- 0.18, significantly lower than that in the control group (0.80 +/- 0.21, P < 0.01), but it was restored rapidly, reaching the normal range on the 25th day, on the 40th day, it became significantly different to the level on the 10th day (P < 0.05) and approached the level in the control group (P > 0.05). No significant pathological change of parotid was found in both groups; but the number of zymogen granules in the RSP group was remarkably more than that in the control group (41.4 +/- 4.9 vs 34.6 +/- 5.2, P < 0.01). Serum level of VIP in the RSP group was significantly less while that of cAMP was higher than that in the control group (22.5 +/- 13.1 mg/L vs 38.5 +/- 14.1 mg/L, and 125.8 +/- 15.5 micromol/L vs 105.3 +/- 16.7 micromol/L, both P < 0.05), but no inter-group difference was found in parotid tissue contents of both VIP and cAMP. All the indices detected became equivalent in the two groups on the 40th day. CONCLUSION: The reduction of salivary protein in Pi-deficient rats induced by RSP may be related to the regulatory pathway of VIP and cAMP.

[Determination of berberine chloride in mouse plasma after intragastric administration of Lianxiang microemulsion].

OBJECTIVE: To determine berberine chloride in mouse plasma for the investigation of the bioavailability of Lianxiang microemulsion in mouse. METHODS: After single dose ig administration,the plasma samples were taken at different time,respectively, for the determination of berberine chloride by HPLC; And the relative bioavailability of Lianxiang microemulsion vs Lianxiang emulsion (reference formulation) was calculated. RESULTS: Berberine chloride was separated well from endogenous foreign substances. The calibration curve of berberine chloride was liner over the range of 10.4-156 microg/L (r = 0.9991) and the method recovery was within 89.6%-94.7%. The intraday RSD or the inter-day RSD < or = 12.9%. After single dose,the AUC0-->24h of the microemulsion and the emulsion were (688.3 +/- 123.7) and (371.4 +/- 68.4) microg x h/L,respectively; And the relative bioavailability of the microemulsion vs the emulsion was (193.1 +/- 63.2)%. CONCLUSION: The validated HPLC method is suitable for the determination of berberine chloride in mouse plasma; And microemulsification can clearly promote the bioavailability in mouse of berberine chloride in Lianxiang prescription.