Alternative Splicing Events in Tumor Immune Infiltration in Colorectal Cancer.

Despite extensive research, the exact mechanisms involved in colorectal cancer (CRC) etiology and pathogenesis remain unclear. This study aimed to examine the correlation between tumor-associated alternative splicing (AS) events and tumor immune infiltration (TII) in CRC. We analyzed transcriptome profiling and clinical CRC data from The Cancer Genome Atlas (TCGA) database and lists of AS-related and immune-related signatures from the SpliceSeq and Innate databases, respectively to develop and validate a risk model of differential AS events and subsequently a TII risk model. We then conducted a two-factor survival analysis to study the association between TII and AS risk and evaluated the associations between immune signatures and six types of immune cells based on the TIMER database. Subsequently, we studied the distribution of six types of TII cells in high- and low-risk groups for seven AS events and in total. We obtained the profiles of AS events/genes for 484 patients, which included 473 CRC tumor samples and 41 corresponding normal samples, and detected 22581 AS events in 8122 genes. Exon Skip (ES) (8446) and Mutually Exclusive Exons (ME) (74) exhibited the most and fewest AS events, respectively. We then classified the 433 patients with CRC into low-risk (n = 217) and high-risk (n = 216) groups based on the median risk score in different AS events. Compared with patients with low-risk scores (mortality = 11.8%), patients with high-risk scores were associated with poor overall survival (mortality = 27.6%). The risk score, cancer stage, and pathological stage (T, M, and N) were closely correlated with prognosis in patients with CRC (P < 0.001). We identified 6479 differentially expressed genes from the transcriptome profiles of CRC and intersected 468 differential immune-related signatures. High-AS-risk and high-TII-risk predicted a poor prognosis in CRC. Different AS types were associated with different TII risk characteristics. Alternate Acceptor site (AA) and Alternate Promoter (AP) events directly affected the concentration of CD4T cells, and the level of CD8T cells was closely correlated with Alternate Terminator (AT) and Exon Skip (ES) events. Thus, the concentration of CD4T and CD8T cells in the CRC immune microenvironment was not specifically modulated by AS. However, B cell, dendritic cell, macrophage, and neutrophilic cell levels were strongly correlated with AS events. These results indicate adverse associations between AS event risk levels and immune cell infiltration density. Taken together, our findings show a clear association between tumor-associated alternative splicing and immune cell infiltration events and patient outcome and could form a basis for the identification of novel markers and therapeutic targets for CRC and other cancers in the future.

Electroacupuncture Versus Pelvic Floor Muscle Training Plus Solifenacin for Women With Mixed Urinary Incontinence: A Randomized Noninferiority Trial.

OBJECTIVE: To evaluate the efficacy and safety of electroacupuncture vs pelvic floor muscle training (PFMT) plus solifenacin for women with mixed urinary incontinence (MUI). PATIENTS AND METHODS: This randomized controlled noninferiority trial was conducted at 10 hospitals in China between March 1, 2014, and October 10, 2016. Participants were randomized 1:1 to receive electroacupuncture (36 sessions) over 12 weeks with 24 weeks of follow-up or PFMT-solifenacin (5 mg/d) over 36 weeks. The primary outcome was percentage change from baseline to week 12 in mean 72-hour incontinence episode frequency (IEF) measured by the 72-hour bladder diary. It was analyzed in the per-protocol set with a prespecified noninferiority margin of 15%. RESULTS: Of 500 women with MUI who were randomized, 467 (239 in the electroacupuncture group and 228 in the PFMT-solifenacin group) completed treatment per protocol and were included in the primary outcome analysis. At weeks 1 through 12, the percentage of reduction from baseline in mean 72-hour IEF was 37.83% in the electroacupuncture group and 36.49% in the PFMT-solifenacin group (between-group difference, -1.34% [95% CI, -9.78% to 7.10%]; P<.001 for noninferiority), which demonstrates noninferiority; the treatment effect persisted throughout follow-up. Statistically significant improvements were found for secondary outcomes in both groups, with no meaningful difference between treatments. CONCLUSION: In women with moderate to severe MUI, electroacupuncture was not inferior to PFMT-solifenacin in decreasing the 72-hour IEF and shows promise as an effective alternative for the treatment of MUI. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT02047032.

Microbiome and metabonomics study of quercetin for the treatment of atherosclerosis.

BACKGROUND: The molecular mechanism of quercetin in the prevention and treatment of AS has been widely reported. However, the microbial and metabolic characteristics of quercetin in AS treatment are still poorly understood. In this study, we aimed to explore the gut microbial and metabolic signatures of quercetin in AS treatment and conduct an integrative analysis on its biomechanism. METHODS: An atherosclerosis mouse model was induced by a high cholesterol diet (HCD). The duration of the quercetin treatment was 12 weeks. We measured TC, TG, HDL and LDL for plasma biochemical analysis and TNF-α and IL-6 for plasma inflammatory analysis. Haematoxylin-eosin (HE) staining was conducted to evaluate the aortic structure and atherosclerosis. Bacterial DNA, which was extracted from mouse faeces, was identified by the V3-V4 regions of the 16S rRNA for microbiological analysis. The HeatMap package of BTtools was applied to visualize the data of the microbial difference matrix according to the OTU results. Fecal metabolites were assessed through LC-MS. Multivariate data analysis was conducted on the normalized data with SIMCA-P+. Significantly different metabolites were extracted based on the Pearson correlation coefficients at the level of P<0.05. Key significantly changed metabolites were screened from the intersection between metabolic signatures of the normal-model and model-quercetin groups. To investigate the biological function of quercetin on AS, we identified the differential metabolic signatures of the model vs. quercetin groups and performed KEGG analyses via MBROLE, MetaboAnalyst database. RESULTS: Quercetin treatment for 12 weeks significantly reduced the levels of TC (P<0.001), TG (P<0.05), HDL (P<0.001), LDL (P<0.001), TNF-α (P<0.001) and IL-6 (P<0.001) compared with the model group. HE staining indicated that quercetin could protect damaged vessels caused by HFD. Bacteroidetes, Firmicutes and Proteobacteria were dominant microbial groups in the samples. There was no significant difference between the three groups (P>0.05) at the phylum level, and the genera Phascolarctobacterium and Anaerovibrio can be regarded as the key microbiota signatures of quercetin treatment. PLS-DA results further showed that these 18 faecal metabolites (clustered in 3 groups) had significant differences between the control, model and quercetin groups throughout the 12-day treatment. According to the quantitative analysis results, 32 key metabolic signatures were screened for quercetin treatment. The main pathway in quercetin treatment is primary bile acid biosynthesis, as 3α,7α,12α,26-tetrahydroxy-5ß-cholestane (C27H48O4) was defined as the most important key metabolic signature. CONCLUSIONS: We explored the gut microbial and metabolic involvement of quercetin in AS treatment and suggest the association between AS and gut metabolic regulation.

Trends in the incidence of childhood type 1 diabetes mellitus in Beijing based on hospitalization data from 1995 to 2010.

OBJECTIVES: To study the incidence trend of type 1 diabetes mellitus (T1DM) in children in Beijing from 1995 to 2010, to compare it with incidences reported worldwide and to predict the requirement of medical resources in the future. METHODS: This study involved newly diagnosed T1DM cases younger than 15 years of age in the Beijing Children's Hospital from January 1995 to December 2010. We calculated the incidence of T1DM children in Beijing according to hospitalization data and Beijing's population. We defined it as the underestimated incidence rate (UE-IR). RESULTS: The UE-IRs of T1DM ranged from around 0.88/100,000 to 2.37/100,000 for children in Beijing younger than 15 years of age from 1995 to 2010. The UE-IR increased faster in boys (1.47 times) and in the age group of 0-4 years (1.89 times) after 2003. The UE-IR was greatest in children aged 5-9 years (1.81/100,000) followed by the age of puberty (10-14 years, 1.76/100,000). The predicted number of new T1DM cases in Beijing will increase 1.97 times over the next 10 years. CONCLUSIONS: The incidence trend of T1DM was increasing gradually in those younger than 15 years of age in Beijing. The incidence of younger children and boys grew faster. The 5- to 14-year-old children represented a high-risk population of T1DM. The number of predicted new T1DM cases will grow rapidly. This means that we should train more health care providers for pediatric diabetes patients, in order to achieve high-quality medical care and to be able to prevent or postpone chronic complications.

Invasive group B streptococcal disease in two pediatric patients with systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is an autoimmune disease associated with high morbidity and mortality, often caused by infection. We report two patients with SLE who were treated with steroids and immunosuppressive medication and then developed invasive Group B Streptococcus (GBS) infections. While GBS infection is rare in the nonneonatal pediatric age group, GBS should be considered when treating SLE patients presenting with signs of infection.

[Quality assessment of randomized controlled trials on wenxin granule for treatment of atrial fibrillation].

OBJECTIVE: To investigate the statement on randomized controlled trials on Wenxin granule for treatment of atrial fibrillation and to judge whether those trials could offer high quality evidence or not, thus improve design level and quality. METHOD: RCTs were searched from home and abroad about atrial fibrillation treated with Wenxin granule, which reported before October, 2010. Jadad scale and CONSORT statement were used. RESULT: There were 66 RCTs retrieved that met inclusion criteria. Using Jadad rating scale, only 2 literatures gain score 4 and 1 literature gains score 3, 54 literatures gain score 2, 7 literatures gain score 1, 2 literatures gain score 0. Only 2 literatures described random number table as the method of grouping. None of the RCTs was reported the allocation concealment. Only 1 literature was used blinding. Fifty-nine literatures were mentioned the lost to follow-up conditions. According to the CONSORT standards, only six literatures (9.1%) mentioned the method of generating the random sequence. Four literatures (6.1%) were quasi-random. Nineteen literatures (28.8%) had inclusion criteria. Six literatures (9.1%) had the follow-up record. Fifty one literatures (77.3%) described the adverse events. None had the estimation of the sample size, intention-to-treat analysis and stratified analysis. None had the ethical approval or informed consent. CONCLUSION: The quality of clinical trials of Wenxin granule in treating atrial fibrillation needs to be improved.

Transforming growth factor-beta1 inhibits steroidogenesis in human trophoblast cells.

Transforming growth factor-beta (TGF-beta) is an important regulator of placental development and function. In this study, we have investigated the effect of TGF-beta1 on steroidogenesis, as well as its sites of action in the steroidogenic pathway by using a choriocarcinoma cell line, JEG-3, and a normal trophoblast cell line (NPC). The effect of TGF-beta1 on progesterone and estradiol production was evaluated in the absence or presence of a membrane-permeable analogue of cholesterol and some intermediate substrates of steroidogenic enzymes. The effect of TGF-beta1 on P450 aromatase (P450arom) mRNA levels was determined by Northern blot analysis. TGF-beta1 significantly decreased progesterone production in both NPC and JEG-3 cells. The inhibitory effect of TGF-beta1 on progesterone production was reversed by addition of 22R-hydroxycholesterol, a membrane-permeable analogue of cholesterol, or pregnenolone. In JEG-3 cells, TGF-beta1 also inhibited estradiol production when androstenedione, but not estrone, was added to the culture. Estradiol production was too low to be detected in NPC cells. Treatment with TGF-beta1 also suppressed aromatase mRNA levels. This study has demonstrated that TGF-beta1 inhibits progesterone and estradiol production by trophoblast cells, and that the sites of TGF-beta1 action on progesterone and estradiol production are likely to be cholesterol transport and P450arom respectively.