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.

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.

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 non-pharmacological interventions for irritable bowel syndrome in adults.

BACKGROUND: Although nonpharmacological interventions (NPI) for irritable bowel syndrome (IBS) have been applied clinically, their relative efficacy and safety are poorly understood. AIM: To compare and rank different NPI in the treatment of IBS. METHODS: Five electronic databases were searched from their inception to January 12, 2020. Data of included publications were analyzed using network meta-analysis (NMA). Quality of endpoints were assessed by tools of the Cochrane Handbook and the GRADEpro software. Pooled relative risk or standardized mean difference with their corresponding 95% confidence intervals were used for statistical analysis. Surface under the cumulative ranking curve (SUCRA) probability value was conducted to rank the examined interventions. Sensitivity analysis was performed to verify the robustness of results and test the source of heterogeneity. RESULTS: Forty randomized controlled trials with 4196 participants were included in this NMA. Compared with routine pharmacotherapies and placebo, acupuncture and cognitive behavioral therapy (CBT) had better efficacy in relieving IBS symptoms. Based on the SUCRA values, acupuncture ranked first in improving overall clinical efficacy and avoiding adverse effects. CBT ranked first in lowering the scores of IBS symptom severity scale, self-rating anxiety scale and self-rating depression scale. CONCLUSION: This study confirmed the efficacy and safety of NPI for improving IBS symptoms, which to some extent recommended several interventions for clinical practice.

Different Traditional Herbal Medicines for the Treatment of Gastroesophageal Reflux Disease in Adults.

BACKGROUND/AIMS: Traditional Herbal Medicines (THM) have been being used for gastroesophageal reflux disease (GERD) for a long time, but clinical evidence is still scarce. We evaluated different THM prescriptions for GERD in adults. METHODS: Data added to nine online databases from their inception to November 30, 2019, were systematically searched. All relevant randomized controlled trials (RCTs) were included and were combined with Bayesian network analysis. The Cochrane Collaboration's risk of bias tool and GRADE profiler version 3.6 were respectively employed to evaluate the quality of evidence of outcomes. RESULTS: Seventeen publications involving 1441 participants were retrieved. The results of our analysis suggested that Jianpi therapy+proton pump inhibitors (PPIs) and Ligan Hewei therapy respectively ranked first in overall clinical efficacy and efficacy under gastroscope; Ligan Hewei therapy+PPIs was the optimum intervention in the improvement of acid regurgitation and heartburn. CONCLUSION: This research indicates that Ligan Hewei therapy and Jianpi therapy, or these therapies separately combined with PPIs, should be recommended as appropriate complementary and alternative treatments based on the specific characteristics of GERD. However, additional well-designed RCTs with high methodological quality are still needed for future research.

Interaction of Cyclooxygenase-2 with Helicobacter pylori Induces Gastric Chronic Nonresolving Inflammation and the Formation of Syndrome of Internal Block of Static Blood in Helicobacter pylori-Related Gastric Diseases.

Cyclooxygenase-2 (COX-2) is an inducible enzyme stimulated by various inflammatory factors (IFs). Chronic gastritis is a classic model of "inflammation-cancer transformation" and Helicobacter pylori-related gastric diseases (HPGD) are specific ones of this model. Traditional Chinese Medicine (TCM) syndromes could play a predictive role in gastric histopathological evolution. To search for early warning evidence about "inflammation-cancer transformation," this study is about to explore interaction of COX-2 with Helicobacter pylori (Hp) in HPGD with different TCM syndromes. All included subjects underwent endoscopy and biopsy. Hp infection was detected by rapid urease test and methylene blue staining. Histopathological characteristics and COX-2 expression in gastric mucosa (GM) were, respectively, observed by hematoxylin-eosin and Elivision™ plus. SPSS 18.0 and Stata 11.0 statistical software packages were used for statistical analysis. Results of immunohistochemical staining in this study showed COX-2 expression in Hp-positive patients was stronger than that in Hp-negative ones. Spearman' analysis indicated that degrees of both Hp infection and COX-2 expression were positively correlated with those of gastric inflammation and inflammatory activity. Compared with the relative normal group, both severe dysplasia group and gastric carcinoma group had more severe Hp infection and COX-2 expression. Compared with the nonsyndrome, syndrome of internal block of static blood (IBSB) had higher scores in semiquantitative analysis of COX-2 protein expression among TCM groups. Moreover, multivariate logistics regression analysis suggested that patients with Hp infection could increase the risk of IBSB. These results indicated that COX-2 interacting with Hp could play an important role in transforming gastric chronic nonresolving inflammation into carcinoma in subjects with HPGD, as well as inducing the formation of IBSB. HPGD together with IBSB could be an early warning evidence for GM with histopathological evolution from benign to malignant.

Efficacy of Jianpi Liqi therapy for functional dyspepsia: A meta-analysis of randomized, positive medicine-controlled trials.

BACKGROUND: We performed this meta-analysis to assess the efficacy and safety of Jianpi Liqi therapy (JLT), a traditional Chinese medicine therapy, in treating functional dyspepsia (FD). METHODS: We systematically searched 13 databases from their inception to 15th, May 2019. Eligible studies were randomized controlled trials (RCTs) that compared JLT medicine with conventional pharmacotherapy (CP) in treating patients with FD. Cochrane Collaboration tool, Review Manager 5.3 and STATA 11.0, GRADE profiler 3.6 were used for evaluating risk of bias, analyzing, and assessing quality of evidence respectively. RESULTS: After exclusions, 15 RCTs including a total of 1451 participants were included for analysis. We found evidence that JLT had better efficacy than CP (domperidone, omeprazole, esomeprazole, mosapride, lansoprazole, compound digestive enzymes, lactasin tablets) for FD (OR 0.34; 95% CI 0.26, 0.45; P < .00001). Moreover, JLT had more improvement on symptoms including abdominal pain, abdominal distention, early satiety, belching, poor appetite, and fatigue compared with CP. In addition, serious adverse events were not observed in treatment courses. CONCLUSION: This meta-analysis suggested that JLT appears to have better efficacy in treating FD compared with CP. It may be an effective and safe therapy option for patients with FD. Though, more large-sample and strictly designed RCTs are needed to confirm our findings.PROSPERO registration number: CRD42019133241.

Polysaccharide extracts of Astragalus membranaceus and Atractylodes macrocephala promote intestinal epithelial cell migration by activating the polyamine-mediated K+ channel.

Astragalus membranaceus (Radix Astragali, RA) and Atractylodes macrocephala (Rhizoma Atractylodis Macrocephalae, RAM) are often used to treat gastrointestinal diseases. In the present study, we determined the effects of polysaccharides extracts from these two herbs on IEC-6 cell migration and explored the potential underlying mechanisms. A migration model with IEC-6 cells was induced using a single-edged razor blade along the diameter of cell layers in six-well polystyrene plates. The cells were grown in control media or media containing spermidine (5 µmol·L-1, SPD), alpha-difluoromethylornithine (2.5 mmol·L-1, DFMO), 4-Aminopyridine (40 µmol·L-1, 4-AP), the polysaccharide extracts of RA or RAM (50, 100, or 200 mg·L-1), DFMO plus SPD, or DFMO plus polysaccharide extracts of RA or RAM for 12 or 24 h. Next, cytosolic free Ca2+ ([Ca2+]cyt) was measured using laser confocal microscopy, and cellular polyamine content was quantified with HPLC. Kv1.1 mRNA expression was assessed using RT-qPCR and Kv1.1 and RhoA protein expressions were measured with Western blotting analysis. A cell migration assay was carried out using Image-Pro Plus software. In addition, GC-MS was introduced to analyze the monosaccharide composition of both polysaccharide extracts. The resutls showed that treatment with polysaccharide extracts of RA or RAM significantly increased cellular polyamine content, elevated [Ca2+]cyt and accelerated migration of IEC-6 cells, compared with the controls (P < 0.01). Polysaccharide extracts not only reversed the inhibitory effects of DFMO on cellular polyamine content and [Ca2+]cyt, but also restored IEC-6 cell migration to control level (P < 0.01 or < 0.05). Kv1.1 mRNA and protein expressions were increased (P < 0.05) after polysaccharide extract treatment in polyamine-deficient IEC-6 cells and RhoA protein expression was increased. Molar ratios of D-ribose, D-arabinose, L-rhamnose, D-mannose, D-glucose, and D-galactose was 1.0 : 14.1 : 0.3 : 19.9 : 181.3 : 6.3 in RA and 1.0 : 4.3 : 0.1 : 5.7 : 2.8 : 2.2 in RAM. In conclusion, treatment with RA and RAM polysaccharide extracts stimulated migration of intestinal epithelial cells via a polyamine-Kv1.1 channel activated signaling pathway, which facilitated intestinal injury healing.

Systematic review and meta-analysis of traditional Chinese medicine in the treatment of constipation-predominant irritable bowel syndrome.

AIM: This meta-analysis analyzed the efficacy and safety of traditional Chinese medicine (TCM) for the treatment of irritable bowel syndrome with constipation (IBS-C). METHODS: We searched seven electronic databases for randomized controlled trials investigating the efficacy of TCM in the treatment of IBS-C. The search period was from inception to June 1, 2017. Eligible RCTs compared TCM with cisapride and mosapride. Article quality was evaluated with the Cochrane Risk Bias Tool in the Cochrane Handbook by two independent reviewers. Begg's test was performed to evaluate publication bias. Review Manager 5.3 and Stata 12.0 were used for analyses. RESULTS: Eleven eligible studies comprising a total of 906 participants were identified. In the primary outcome, TCM showed significant improvement in overall clinical efficacy compared with cisapride and mosapride (odds ratio [OR] = 4.00; 95% confidence interval [CI]: 2.74,5.84; P < 0.00001). In terms of secondary outcomes, TCM significantly alleviated abdominal pain (OR = 5.69; 95% CI: 2.35, 13.78; P = 0.0001), defecation frequency (OR = 4.38; 95% CI: 1.93, 9.93. P = 0.0004), and stool form (OR = 4.96; 95% CI: 2.11, 11.65; P = 0.0002) in the treatment group as compared to the control group. A lower recurrence rate was associated with TCM as compared to cisapride and mosapride (OR = 0.15; 95% CI: 0.08, 0.27; P < 0.00001). No adverse effects were observed during TCM treatment. CONCLUSIONS: TCM showed greater improvement in terms of clinical efficacy in the treatment of IBS-C than cisapride and mosapride, although it was not possible to draw a definitive conclusion due to the small sample size, high risk, and low quality of the studies. Large multi-center and long-term high-quality randomized control trials are needed.

[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.

Atractylodes macrocephala Koidz stimulates intestinal epithelial cell migration through a polyamine dependent mechanism.

ETHNOPHARMACOLOGICAL RELEVANCE: Atractylodes macrocephala Koidz (AMK), a valuable traditional Chinese herbal medicine, has been widely used in clinical practice for treating patients with disorders of the digestive system. AMK has shown noteworthy promoting effect on improving gastrointestinal function and immunity, which might represent a promising candidate for the treatment of intestinal mucosa injury. The aim of this study was to investigate the efficacy of AMK on intestinal mucosal restitution and the underlying mechanisms via intestinal epithelial (IEC-6) cell migration model. MATERIALS AND METHODS: A cell migration model of IEC-6 cells was induced by a single-edge razor blade along the diameter of the cell layers in six-well polystyrene plates. After wounding, the cells were grown in control cultures and in cultures containing spermidine (5µM, SPD, reference drug), alpha-difluoromethylornithine (2.5mM, DFMO, polyamine inhibitor), AMK (50, 100, and 200mg/L), DFMO plus SPD and DFMO plus AMK for 12h. The polyamines content was detected by high-performance liquid chromatography (HPLC) with pre-column derivatization. The Rho mRNAs expression levels were assessed by Q-RT-PCR. The Rho and non-muscle myosin II proteins expression levels were analyzed by Western blot. The formation and distribution of non-muscle myosin II stress fibers were monitored with immunostaining techniques using specific antibodies and observed by confocal microscopy. Cell migration assay was carried out using inverted microscope and the Image-Pro Plus software. All of these indexes were used to evaluate the effectiveness of AMK. RESULTS: (1) Treatment with AMK caused significant increases in cellular polyamines content and Rho mRNAs and proteins expression levels, as compared to control group. Furthermore, AMK exposure increased non-muscle myosin II protein expression levels and formation of non-muscle myosin II stress fibers, and resulted in an acceleration of cell migration in IEC-6 cells. (2) Depletion of cellular polyamines by DFMO resulted in a decrease of cellular polyamines levels, Rho mRNAs and proteins expression, non-muscle myosin II protein formation and distribution, thereby inhibiting IEC-6 cell migration. AMK not only reversed the inhibitory effects of DFMO on the polyamines content, Rho mRNAs and proteins expression, non-muscle myosin II protein formation and distribution, but also restored cell migration to control levels. CONCLUSIONS: The results obtained from this study revealed that AMK significantly stimulates the migration of IEC-6 cells through a polyamine dependent mechanism, which could accelerate the healing of intestinal injury. These findings suggest the potential value of AMK in curing intestinal diseases characterized by injury and ineffective repair of the intestinal mucosa in clinical practice.

Atractylodes macrocephala Koidz promotes intestinal epithelial restitution via the polyamine--voltage-gated K+ channel pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Atractylodes macrocephala Koidz (AMK) has been used widely as a digestive and tonic in traditional Chinese medicine. AMK has shown noteworthy promoting effect on intestinal epithelial cell migration, which might represent a promising candidate for the treatment of intestinal mucosa injury. The aim of this study was to investigate the efficacy of AMK on intestinal mucosal restitution and the underlying mechanisms via IEC-6 cell migration model. MATERIALS AND METHODS: A wounding model of IEC-6 cells was induced by a single-edge razor blade along the diameter of six-well polystyrene plates. The cells were grown in control cultures and in cultures containing spermidine (5 µmol/L, SPD, reference drug), alpha-difluoromethylornithine (2.5 mmol/L, DFMO, polyamine inhibitor), AMK (50, 100, and 200 µg/mL), DFMO plus SPD and DFMO plus AMK for 24h. The membrane potential (MP) and cytosolic free Ca(2+) concentration ([Ca(2+)]cyt) were detected by flow cytometry, and polyamines content was determined via high-performance liquid chromatography (HPLC). The expression of Kv1.1 mRNA and protein levels were assessed by RT-qPCR and Western blot analysis, respectively. Cell migration assay was carried out using the Image-Pro Plus software. All of these indexes were used to evaluate the effectiveness of AMK. RESULTS: (1) Treatment with AMK caused significant increases in cellular polyamines content, membrane hyperpolarization, an elevation of [Ca(2+)]cyt and an acceleration of cell migration in IEC-6 cells, as compared to control group. (2) AMK not only reversed the inhibitory effects of DFMO on the polyamines content, MP, and [Ca(2+)]cyt but also restored IEC-6 cell migration to control levels. (3) The Kv1.1 mRNA and protein expression were significantly increased by AMK treatment in control and polyamine-deficient IEC-6 cells. CONCLUSIONS: The results of our current studies revealed that treatment with AMK significantly stimulates the migration of intestinal epithelial cells through polyamine-Kv1.1 channel signaling pathway, which could promote the healing of intestinal injury. These results suggest the potential usefulness of AMK to cure intestinal disorders characterized by injury and ineffective repair of the intestinal mucosa.

[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.

[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.

[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.

[Effects of Atractylodes macrocephala monosaccharide composition on cytodifferentiation and villin expression of IEC-6 cells in vitro].

OBJECTIVE: To study the influence of Atractylodes macrocephala monosaccharide composition (AMMC) on IEC-6 cells by observing cytodifferentiation and expression of villin. METHOD: Observed cell morphous under light microscope, observed cell ultramicrostructure under transmission electron microscope, evaluated villin mRNA by way of RT-PCR, and detected protein of villin by immunofluorescence. Cells were divided into the groups including blank control,positive control (given with gastrin 250 microg/L) and AMMC groups (given with AMMC 62.5, 125, 250, 500, 1000 mg/L respectively). RESULTS: 1. Cells in blank control group were immature under light microscope, which were in state of differentiation in groups with gastrin and different concentration of AMMC. 2. Cells in blank control group had bigger nucleuses with higher ratio of heterochromatin and much less villin at the edge of cells, which in groups with gastrin and different concentration of AMMC had smaller nucleuses with normal ratio of heterochromatin/heterochromatin and abundance of villin at the edge of cells. 3. Villin mRNA of IEC-6 cells in groups with gastrin and different concentration of AMMC were obviously much more than that of cells in blank control group 6 h afeer treatment. 4. Little protein of villin was detected by immunofluorescence in blank control group, which was much brighter in group with gastrin, and less bright in group with different concentration of AMMC, in which specific villin assembled. CONCLUSION: AMMC can promote cytodifferentiation by up-regulating the expression and distribution of villin in IEC-6 cells.

[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.

[Screening of anti-diarrhea effective fractions from guava leaf].

OBJECTIVE: To screen the fractions from guava leaf to ascertain the effective anti-diarrhea fraction and to establish the method of quality control. METHODS: A series of samples from guava leaf were prepared through extraction and separation by solvents progressively, and their effect on the mice diarrhea models induced by senna decoction were tested individually; and the constituent correlated with the effect of the samples on the models was determined by HPLC. RESULTS: The effect of the fractions with moderate-polar glucosides from guava leaf was better than that of the other fractions; the effect of the fractions with the glucosides was correlated with the content of the quercetin glucosides in the fractions. CONCLUSION: The fractions with the moderate-polar glucosides is the effective anti-diarrhea fraction of guava leaf; determination of the quercetin glucosides can be used for quality control of guava leaf and its extracts.

[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.

[Study on self-microemulsifying drug delivery system of LianXiang prescription].

OBJECTIVE: To prepare self-microemulsifying drug delivery system (SMEDDS) of LianXiang prescription. METHODS: After the test of solubility for the initial screening, surfactants and cosurfactants with certain oil phase were screened through the study of ternary phase diagram and in accordance with the size of self-microemulsifying areas; To screen and optimize the self-microemulsifying formulation, the influence of related factors such as the ingredients of the prescription and dilution media on the self-microemulsifying areas were tested, and the particle dimension of microemulsion was determined. The tests of self-microemulsifying rate and stability were carried out to evaluate the formulation. RESULTS: The total alkaloid of Coptis chinensis--the essential oil of Eugenia caryophyllata-Cremophor RH60-1,2-propylene glycol(40: 20: 60: 20, w/w) was the optimal formulation; it had good self-microemulsifying efficiency and stability, and its microemulsion had smaller particle dimension and polydispersity index (17.4 nm, 0.176). CONCLUSION: The optimal formulation can be used as SMEDDS of LianXiang prescription for follow-up researches.