Exploring the Mechanism of Indigo Naturalis in the Treatment of Ulcerative Colitis Based on TLR4/MyD88/NF-κB Signaling Pathway and Gut Microbiota

Background: Clinical trials have proven that indigo naturalis is a candidate drug for treating ulcerative colitis (UC), but its therapeutic mechanism is still unclear.Purpose: This study aimed to evaluate the protective effect and mechanism of indigo naturalis to treat mice with dextran sulfate sodium (DSS)-induced UC.Methods: DSS-induced UC mice were treated with indigo naturalis (200 mg/kg), indigo (4.76 mg/kg), and indirubin (0.78 mg/kg) for 1 week. The anti-UC mechanism of indigo naturalis was studied by pathological section, inflammatory factor, western blot, and 16S rRNA sequencing.Results: According to body weight change, disease activity index, and colon length, indigo naturalis had the strongest anti DSS-induced UC effect, followed by indirubin and indigo. Pathological section showed that indigo naturalis, indigo, and indirubin could reduce the infiltration of inflammatory cells, increase the secretion of intestinal mucus, and repair the intestinal mucosa. Indigo naturalis, indigo, and indirubin could reduce IL-1β,IL-6, and TNF-α by inhibiting TLR4/MyD88/NF-κB signal transduction. Indigo naturalis and indigo could also reduce IgA and IgG both in serum and colon tissue. In addition, indigo naturalis, indigo, and indirubin could adjust the gut microbiota structure of DSS-induced UC mice, reducing the ratio of Firmicutes/Bacteroidetes and increasing the abundance of probiotics.Conclusion: Indigo and indirubin are one of the main anti-UC components of indigo naturalis. INN could regulate intestinal flora, reduce inflammation, repair intestinal mucosa, and improve the physiological status of DSS-induced UC mice and its anti-UC mechanism may be involved in inhibiting TLR4/MyD88/NF-κB signal transduction.

Teratogenic Toxicity Evaluation of Bladder Cancer Specific Oncolytic Adenovirus on Mice

Background: In our previous studies, we had demonstrated the efficiency and specificity of constructed bladder tissue specific adenovirus Ad-PSCAE-UPII-E1A-AR (APU-EIA-AR) on bladder cancer, we also investigated the virus biodistribution and body toxicity in nude mice. However, the safety of the bladder cancer specific oncolytic adenovirus on fetal mice and F1 mice should be under intense investigation. Objectives: In order to evaluate the teratogenic toxicity of bladder cancer specific oncolytic adenovirus APU-EIA-AR on mice, in this study, we investigated the fetal mice weight, fetal body length and tail length, fetal skeleton development, as well as the F1 mice weight, growth curve, and major organ pathology. These teratogenic toxicity data of bladder tissue specific adenovirus AdPSCAE-UPII-E1A-AR (AD) would provide safe information prior to embarking on clinical trials. Methods: At sixth day of being fertilized, the pregnant mice began to be intramuscular administrated with AD (1×107VP, 1×108VP, 1×109VP) every other day for ten days. Then ,the pregnants were devided into two groups. One group was euthanized at seventeenth day, took out the fetal mice ,observing the bone structure of the infants. The oth er group was observed until natural childbirth. The Filial Generation (F1) are feeded and growth for 30 days, the variation in the growth progress and developmen t were assessed. Then the mice were euthenazied, the organ tissue were performed pathological section and HE staining, observing the changes under microscope. Results: In the process of teratogenic toxicity test , comparing with control group ,the Placenta weight ,fetal mice weight , body len gth and tail length of mice fetal in adenovirus treated group did not reveal any alteration. Comparing with PBS group, there is no obvious change in skeleton of fetal mice treated with adenovirus. During the development process of F1 mice treated with adenovirus, the changes of mice weight show statistical significance. Howerer, in the progress of growth curve, this difference is not very obvious. Furthermore, the pathological section showed no obvious alteration in major organs. Conclusion: Our study demonstrated that bladder cancer specific adenovirus Ad-PSCAE-UPII-E1A-AR appear safe in the pregnant mice without any discernable effects on fetal mice and F1 development. Hence, It is a relatively safe for tumor gene therapy.

Study on the mechanism of Indigo naturalis and its two main compounds (indigo and indirubin) in the treatment of ulcerative colitis based on TLR4/MyD88/NF-κB pathway and intestinal microorganisms

Background: Indigo naturalis is a natural dye extracted from plants and has a good anti-inflammatory effect. Clinical studies have shown that it can improve ulcerative colitis (UC), but the active constituents and the mechanism are unclear. Methods: The anti-UC activity of Indigo naturalis and its two main compounds (indigo and indirubin) were investigated in dextran sulfate sodium (DSS)-induced UC mice. Indigo naturalis, indigo and indirubin were administrated to DSS-induced UC rats by oral gavage for 1 weeks. The anti-UC effect was evaluated by pathological section, inflammatory cytokine production, western blotting, and gut microbiota analysis via 16S rRNA sequencing. Results: Indigo naturalis, indigo and indirubin can improve the UC induced by DSS. Their effect intensity is Indigo naturalis > indirubin > indigo based on disease activity index, body weight, colon length and pathological section. Indigo naturalis, indigo and indirubin also decrease the expression of NF-κB，TLR4 and MYD88 proteins, thus reducing the level of related inflammation cytokines (IL-1β, IL-6 and TNF-α) both in serum and tissue. In addition, Indigo naturalis and indigo improved symptoms of gut microbial disturbance, and decreased Firmicutes/Bacteroidetes ratio and the significantly increased probiotics such as Lactobacillus. Indirubin has little effect on the regulation of gut microbial. Conclusions: Indigo naturalis could attenuate the DSS-induced UC in mice, by means of ameliorating intestinal inflammation, improving intestinal mucosa, and regulating the disturbed gut microbiota. Indigo and indirubin could also attenuate the DSS-induced UC in mice, but their comprehensive effect is not as good as Indigo naturalis.

P.033 Novel presentation of NMDA receptor encephalitis

Background: NMDA receptor encephalitis (NMDARE) is associated with pre-existing psychiatric symptoms and seizure disorders. It is not typically associated with elevated ICP. Diagnostically, EEG findings in NMDARE are characteristic as are the pathological features of ovarian teratomas associated with this disease. We report a patient who tested positive for NMDARE however presented with features not known to be associated with the disease including elevated ICP, atypical EEG findings and grossly atypical features on pathological section. Results: A 26 year old woman presented with psychiatric symptoms and status epilepticus. On examination, she was found to have papilledema and eleveated ICP on measurement. Her imaging and EEG demonstrated atypical findings, not consistent with NMDARE. CT scan of the abdomen demonstrated an adnexal mass. CSF studies eventually tested positive for NMDARE and following removal of her ovarian teratoma, the pathology demonstrated atypical findings for lesions associated with NMDARE classically. Conclusions: NMDARE is a new entity, which has historically shown a typical clinical course. Our case demonstrates a previously undescribed presentation of NMDARE with elevated ICP, atypical EEG findings and unique pathology of the associated ovarian teratoma.

Pathology Changes of Kidney in Mouse Following DBP&DEHP Exposure

The toxic effects of di-2-ethylhexyl phthalate（DEHP）and di-n-butyl phthalate（DBP）on the kidney of mouse was studied. Forty mice were randomly divided into 4 groups , 10 mice in each group. There were three experimental groups:DBP+DEHP（2.5g/kgDEHP+0.5g/kgDBP）,DEHP(2.5g/kg）,DBP（0.5g/kg）and a control group which was given coin oil. The experiment was conducted through gavage every other day for 35 days. The organ coefficients were counted and pathological changes in kidney were observed. The weights of mice of each group increased with the time,but there were no difference between each group in statistic. The organ coefficients of kidney of DEHP and DBP&DEHP group were significantly higher than that of DBP group and control group. Infected groups appeared injury to some extend by observing the pathological section. It reveals DBP and DEHP have toxicity on kidney.

The Using of Combination Algorithm to the Gastric Carcinoma Nucleus Segmentation

The nucleus segmentation of gastric carcinoma pathological section microscopic image is a challenge. Watershed and GVF model can be applied to ordinary nucleus segmentation, but each has its advantages and disadvantages. Combine two algorithms and pathology expertise. The initial contours of nucleus are achieved by watershed transformation first, and then the final contours are approaching by GVF model. Experiments show that the combined algorithm is valid.