The effects of Wumeiwan (WMW) on TNF-alpha, IL-6, IL-8, IL-10 and NF-kappaBp65 in rats with ulcerative colitis (UC) were investigated, the curative effectiveness of WMW vs salicylazosulfapyridine (SASP) was compared, and the action mechanism was analyzed. Fifty-Six Sprague-Dawley (SD) rats were randomly divided into four groups (n=14 in each group, with equal ratio of male and female): normal control group, model group, SASP group, and WMW group. Except normal control group, the rat UC models in the remaining three groups were established using the method of 2.4-dinitrochlorobenzene (DNCB) immunization and acetic acid local enema. The rats in model group, SASP group, and WMW group were treated with distilled water, SASP, and WMW respectively. The changes in the symptoms and signs were observed, and levels of IL-6, IL-8, TNF-alpha, IL-10 and the expression of NF-kappaBp65 in the colonic tissues were statistically analyzed. The results showed that the levels of IL-6, IL-8, and TNF-alpha were significantly increased (P<0.01), while those of IL-10 significantly reduced (P<0.01) after establishment of rat UC models as compared with normal control group. The levels of IL-6, IL-8, and TNF-alpha were obviously lower, but the level of IL-10 was obviously higher in WMW and SASP groups than those in model group (P<0.05). The levels of IL-6, IL-8, and TNF-alpha were lower, while the level of IL-10 was higher in WMW group than in SASP group. NF-kappaBp65 was expressed negatively or weakly in normal colonic tissues. The positive expression rate of NF-kappaBp65 in WMW group and SASP group was obviously lower than in model group (P<0.01), and there was significant difference between WMW group and SASP group (P<0.05). It was concluded that rat UC model was established successfully. WMW could up-regulate the expression of IL-10, down-regulate the expression of TNF-alpha, IL-6, IL-8, and inhibit the NF-kappaBp65 activity to adjust immune function, indicating WMW had better curative effects on UC in rats.

OBJECTIVETo investigate the pathogenetic mechanism of beta-arrestin1 in the rat's experimental colitis, whether the delta opioid receptor-beta-arrestin1 -Bcl-2 signal transduction pathway involves the pathological process of experimental colitis in rats, and whether oxymatrine could attenuate colitis through this pathway.

METHODSTwenty-six SD rats were randomly divided into four groups, the normal control group, the model group, the mesalazine treated group and the oxymatrine treated group (8 rats in the last group and 6 each in the others). The colitis model was established with trinitrobenzene sulfonic acid (TNBS), and rats in the latter two groups were treated by oxymatrine (intramuscular injection) and mesalazine (3 mL solution gavaged) for 15 days, respectively, while rats in the former two groups were fed with equal volume of distilled water. Symptoms of diarrhea and bloody stool as well as colonic patho-histologic changes were observed, and changes in expressions of delta opioid receptor, beta-arrestin1 and Bcl-2 in rat's colon tissue and spleen T lymphocytes were detected with immuno-histochemistry and Western immune-blotting techniques, respectively.

RESULTSIn contrast to the normal control group, expressions of delta opioid receptor, beta-arrestin1 and Bcl-2 were significantly higher in the model group (P < 0.01); compared with the model group, they were significantly lower in the two treated groups (P < 0.01).

CONCLUSIONSDelta opioid receptor-beta-arrestin1 -Bcl-2 signal transduction pathway participates in the pathogenesis of TNBS-induced experimental colitis in rats. Oxymatrine can intervene the signal transduction, which may be one of the mechanisms of oxymatrine in attenuating colitis in rats.

Alkaloids ; pharmacology ; Animals ; Arrestins ; metabolism ; Colitis ; chemically induced ; etiology ; physiopathology ; Colon ; drug effects ; Inflammatory Bowel Diseases ; etiology ; physiopathology ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Quinolizines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, delta ; metabolism ; Signal Transduction ; Trinitrobenzenesulfonic Acid ; beta-Arrestins

Objective To study the clinical characteristics of autism in febrile seizures plus (FS+) and the relationship between autism and SCN1A mutation. Methods Clinical data of 103 patients with FS+ treated in epilepsy centre of the Second Affiliated Hospital of Guangzhou Medical University were collected and analyzed. According to the international criteria, generalized epilepsy with febrile seizures plus (GEFS+), partial seizures with febrile seizures plus (PEFS+), Dravet syndrome (DS) and autism were diagnosed. Genomic DNA was obtained from blood samples. SCN1A were PCR amplified and mutations were detected by DHPLC and sequencing. Result Mental retardation was found in 53.8%of patients with GEFS+, 69.2%of patients with PEFS+, and all patients with DS, respectively. One in GEFS+, one in PEFS+and nine in DS patients were accompanied with autism (P<0.01). Among FS+patients with autism, one SCN1A mutation was found in PEFS+patients, while six SCN1A mutations were found in DS patients. Conclusions Majority of GEFS+and PEFS+patients showed mental retardation, while all the DS patients were accompanied with retardation. The occurrence of autism with DS is higher than GEFS+and PEFS+. No definite relationship between autism and SCN1A mutation was indicated.

RHO-related GTPases of plants (ROPs) are a class of signal transduction G proteins (alsoknown as GTP binding proteins) widely existing in plants. ROP proteins act as " molecular switches" toregulate the signal transduction process during cellular activities such as plant cell polarity regulation, plant morphological development, hormone level regulation, stress responses and many other life activitiesby shifting between inactive GDP-binding and active GTP-binding forms in the cells. In this review, thedomain structure, classification, the mechanism of activity regulation and biological functions of ROPproteins were summarize. Furthermore, ROP proteins from Arabidopsis, maize, rice and barley werephylogenetically analyzed. The results show that ROP proteins were classified into two types based on thedomain structure of the proteins. However, these ROP proteins were divided into 4 clades based on thesimilarity of protein sequences. Furthermore, the mechanism of ROP proteins as a molecular switchregulating various signaling pathways in cells, and the specific functions and mechanisms of ROPs in thepolarized growth of pollen tubes, root hairs and plant pavement cells and other stress responses werecharacterized. In addition, the research progress of the function of ROPs in plant hormones such as ABA, IAA and BR mediated signal transduction were described as well. At last, the unanswered questions suchas why different ROP proteins play distinct roles in the same signaling pathway and how ROPs coordinatedifferent signal pathways to jointly regulate a plant’ s development or physiological process werediscussed, which may shed light on future research.

OBJECTIVETo explore the effect and its mechanism of acupoint catgut-embedding for experimental colitis in rats.

METHODSEighteen SD rats were randomly divided into a normal control (NC) group, a model (MO) group and a acupoint catgut-embedding (AC) group, 6 rats in each group. Animals in MO group and AC group were treated by trinitro-benzene-sulfonic acid (TNBS) to establish model with colitis. No other treatment was given to the rats in MO group, but acupoint catgut-embedding was implanted at "Shangjuxu" (ST 37), "Tianshu" (ST 25) and "Dachangshu" (BL 25) in the rats in AC group. The symptoms of diarrhea and bloody stool, and histopathology changes in colon were observed 15 days after the treatment. Expression of IL-17, beta2AR, NFkappaBp65 were observed by immunohistochemistry. Expressions of NF-kappaBp65 and beta2AR in splenic lymphocyte were detected by the Western blot method.

RESULTSDiarrhea and mucus bloody purulent stool were soon controlled, and colon mucosa injures were obviously improved in AC group. The NF-kappaBp65 value in splenic lymphocytes of 249.70 +/- 13.66 in MO) group was higher than 86.22 +/- 8.09 in NC group (P < 0.01), and 219.02 +/- 7.42 in AC group was less than that in MO group (P < 0.01). The expression of beta2AR in splenic lymphocytes of 594.97 +/- 173.22 in MO group was less than 957.45 +/- 171.56 in NC group (P < 0.01), and 1335.93 +/- 244.34 in AC group was higher than that in MO group (P < 0.01). The expression of IL-17 in colon mucosa in MO group was increased, while the expression of IL-17 in colon mucosa in AC group was decreased.

CONCLUSIONAcupoint catgut-embedding at Shangjuxu (ST 37), Tianshu (ST 25) and Dachangshu (BL 25) has obviously effect in treating experimental colitis and the mechanism may be related to regulate the expression of IL-17, beta2AR and NF-kappaBp65.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Catgut ; Colitis ; genetics ; metabolism ; therapy ; Disease Models, Animal ; Female ; Gene Expression Regulation ; Humans ; Interleukin-17 ; Male ; NF-kappa B ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Androgen ; genetics ; metabolism

Background: The initiation of sodium-glucose cotransporter-2 inhibitors (SGLT2i) typically leads to a reversible initial dip in estimated glomerular filtration rate (eGFR). The implications of this phenomenon on clinical outcomes are not well-defined. Methods: We searched MEDLINE, Embase, and Cochrane Library from inception to March 23, 2023 to identify randomized controlled trials and cohort studies comparing kidney and cardiovascular outcomes in patients with and without initial eGFR dip after initiating SGLT2i. Pooled estimates were calculated using random-effect meta-analysis. Results: We included seven studies in our analysis, which revealed that an initial eGFR dip following the initiation of SGLT2i was associated with less annual eGFR decline (mean difference, 0.64; 95% confidence interval [CI], 0.437 to 0.843) regardless of baseline eGFR. The risk of major adverse kidney events was similar between the non-dipping and dipping groups but reduced in patients with a ≤10% eGFR dip (hazard ratio [HR], 0.915; 95% CI, 0.865 to 0.967). No significant differences were observed in the composite of hospitalized heart failure and cardiovascular death (HR, 0.824; 95% CI, 0.633 to 1.074), hospitalized heart failure (HR, 1.059; 95% CI, 0.574 to 1.952), or all-cause mortality (HR, 0.83; 95% CI, 0.589 to 1.170). The risk of serious adverse events (AEs), discontinuation of SGLT2i due to AEs, kidney-related AEs, and volume depletion were similar between the two groups. Patients with >10% eGFR dip had increased risk of hyperkalemia compared to the non-dipping group. Conclusion Initial eGFR dip after initiating SGLT2i might be associated with less annual eGFR decline. There were no significant disparities in the risks of adverse cardiovascular outcomes between the dipping and non-dipping groups.

OBJECTIVETo investigate the β2-adrenoceptor (β2AR)-β-arrestin2-nuclear factor-κB (NF-κB) signal transduction pathway and the intervention effects of oxymatrine in a rat model of ulcerative colitis.

METHODSForty SD rats were randomly divided into four groups, which included the normal control group, the model group, the mesalazine group and the oxymatrine treatment group, with 10 rats per group. Experimental colitis induced with trinitrobenzene sulfonic acid (TNBS) was established in each group except the normal control group. The rats in the oxymatrine treatment group were treated with intramuscular injection of oxymatrine 63 mg/(kg·d) for 15 days and the rats in the mesalazine group were treated with mesalazine solution 0.5 g/(kg·d) by gastric lavage for 15 days. The rats in the normal control group and model group were treated with 3 mL water by gastric lavage for 15 days. Diarrhea and bloody stool were carefully observed. Histological changes in colonic tissue were examined on day 7 in 2 rats per group that were randomly selected. The expression of β2AR, β-arrestin2 and NF-κB p65 in colon tissue and spleen lymphocytes were detected with immunohistochemistry and Western immunoblotting techniques on day 16 after fasting for 24 h. Six rats died of lavage with 2 each in the normal control, the model group and the mesalazine group; and were not included in the analysis.

RESULTSThe rats in the model group suffered from looser stool and bloody purulent stool after modeling. But in the oxymatrine and mesalazine groups, looser stool and bloody purulent stool reduced after treatment. And the colonic wall in the model group was thickened and the colon length shortened. The colon mucosa was congested in multiple areas with edema, erosion, superficial or linear ulcer and scar formation, while the intestinal mucosa injury reduced in the mesalazine and oxymatrine groups (P<0.01). In colonic mucosa and in spleen lymphocytes, compared with the normal control group, the expression of NF-κBp65 were significantly increased (P<0.01) in the model group while the expressions of β 2AR and β-arrestin2 were significantly decreased (P<0.01). Compared with the model group, the expression of NF-κ Bp65 was significantly decreased in the mesalazine group (P<0.01) and oxymatrine treatment group (P<0.01) while the expressions of β2AR and β-arrestin2 were significantly increased (P<0.01). There were no statistically significant differences in the expression of β2AR, β-arrestin2 and NF-κBp65 between the mesalazine group and oxymatrine group (P>0.05).

CONCLUSIONSThe β2AR-β-arrestin2-NF-κB signal transduction pathway participated in the pathologic course of ulcerative colitis. Oxymatrine attenuated ulcerative colitis through regulating the β2AR-β-arrestin2-NF-κB signal transduction pathway.

Alkaloids ; pharmacology ; Animals ; Arrestins ; metabolism ; Colitis, Ulcerative ; drug therapy ; metabolism ; Colon ; drug effects ; pathology ; Intestinal Mucosa ; drug effects ; metabolism ; pathology ; Lymphocytes ; metabolism ; pathology ; Male ; NF-kappa B ; metabolism ; Quinolizines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Adrenergic, beta-2 ; metabolism ; Signal Transduction ; drug effects ; Spleen ; pathology ; beta-Arrestins

The effects of Wumeiwan (WMW) on TNF-α, IL-6, IL-8, IL-10 and NF-κBp65 in rats with ulcerative colitis (UC) were investigated, the curative effectiveness of WMW vs salicylazosul-fapyridine (SASP) was compared, and the action mechanism was analyzed. Fifty-Six Spra-gue-Dawley (SD) rats were randomly divided into four groups (n=14 in each group, with equal ratio of male and female): normal control group, model group, SASP group, and WMW group. Except normal control group, the rat UC models in the remaining three groups were established using the method of 2.4-dinitrochlorobenzene (DNCB) immunization and acetic acid local enema. The rats in model group, SASP group, and WMW group were treated with distilled water, SASP, and WMW re-spectively. The changes in the symptoms and signs were observed, and levels of IL-6, IL-8, TNF-α,IL-10 and the expression of NF-κBp65 in the colonic tissues were statistically analyzed. The results showed that the levels of IL-6, IL-8, and TNF-α were significantly increased (P<0.01), while those of IL-10 significantly reduced (P<0.01) after establishment of rat UC models as compared with normal control group. The levels of IL-6, IL-8, and TNF-α were obviously lower, bat the level of IL-10 was obviously higher in WMW and SASP groups than those in model group (P<0.05). The levels of IL-6,IL-8, and TNF-α were lower, while the level of IL-10 was higher in WMW group than in SASP group.NF-κBp65 was expressed negatively or weakly in normal colonic tissues. The positive expression rate of NF-κBp65 in WMW group and SASP group was obviously lower than in model group (P<0.01), and there was significant difference between WMW group and SASP group (P<0.05). It was concluded that rat UC model was established successfully. WMW could up-regulate the expres-sion oflL-10, down-regulate the expression ofTNF-α, IL-6, IL-8, and inhibit the NF-κBp65 activity to adjust immune function, indicating WMW had better curative effects on UC in rats.

Objective To study the influence of SCNIA intronic mutations in mRNA splicing in febrile seizures related epilepsy,and investigate the association between splicing changes and genotype-phenotype-inheritance pattern.Methods Molecular cloning of 5 SCN1A intronic mutations was performed in patients with partial epilepsy with antecedent febrile seizures plus (PEFS+) and Dravet syndrome (DS) through constructing mutant and wild-type plasmids of pTragetE2-3-4-5 and E24-25-26 by using Minigene splicing assay,and the in vitro expressions in HENK293 cells were detected.The mRNA splicing changes were analyzed qualitatively and quantitatively by reverse transcription (RT)-PCR and real time quantitative (q)-PCR.Results (1) Using RT-PCR,DS mutants presented a whole exon skipping without significant remain of normal mRNA transcripts,while PEFS+ mutants showed partial exon skipping or intronic insertion with coexistence of normal and aberrant mRNA transcripts.(2) Statistical differences were found between relative quantity (RQ) of aberrant and normal mRNA in PEFS+ mutant (c.473+5G＞A:4.92％±1.05％ and 6.10％±0.21％;c.473+5G＞C:7.97％±1.12％ and 3.94％ ±1.25％) and that in DS mutant (c.602+1G＞A:60.51％±1.81％ and 0.060％±0.022％,P＜0.05);similarly,there were statistical differences between relative RQ of normal and aberrant mRNA in PEFS+ mutant c.4853-25T＞A (71.22％±11.92％ and 7.38％±1.61％) and that in DS mutant c.4853-1G＞C (0.08％±0.01％ and 22.11％±2.83％,P＜0.05).Conclusion The position and difference of splicing patterns of SCNIA intronic mutations are potential molecular pathogenesis for phenotypic difference of febrile seizures related epilepsy.

OBJECTIVETo improve Luo-Ye pump-based stress-forming system and optimize the stimulating effect on smooth muscle cells during cultivation of tissue-engineered blood vessels (TEBV).

METHODSA new Luo-Ye pump-based TEBV 3D culture system was developed by adding an air pump to the output of the bioreactor. A pressure guide wire was used to measure the stress at different points of the silicone tube inside the TEBV bio-reactor, and fitting curves of the stress changes over time was created using Origin 8.0 software. The TEBVs were constructed by seeding vascular smooth muscle cells (VSMCs) isolated from human umbilical artery on polyglycolic acid (PGA) and cultured under dynamic conditions with 40 mmHg resistance (improved group), dynamic conditions without resistance (control group) or static condition (static group) for 4 weeks. The harvested TEBVs were then examined with HE staining, masson staining, α-SMA immunohistochemical staining, and scanning and transmission electron microscopy with semi-quantitative analysis of collagen content and α-SMA expression.

RESULTSThe measured stress values and the fitting curves showed that the stress stimuli from the Luo-Ye pump were enhanced by adding an air pump to the output of the bioreactor. Histological analysis revealed improved VSMC density, collagen content and α-SMA expression in the TEBVs constructed with the improved method as compared with those in the control and static groups.

CONCLUSIONAdding an air pump to the Luo-Ye pump significantly enhances the stress stimulation in the TEBV 3-D culture system to promote the secretion function of VSMCs.

Bioreactors ; Blood Vessel Prosthesis ; Cells, Cultured ; Collagen ; metabolism ; Humans ; Myocytes, Smooth Muscle ; cytology ; Polyglycolic Acid ; Tissue Engineering ; methods