Artificial cavernosa-like tissue based on multibubble Matrigel and a human corpus cavernous fibroblast scaffold.

Ex vivo tissue culture of the human corpus cavernosum (CC) can be used to explore the tissue structural changes and complex signaling networks. At present, artificial CC-like tissues based on acellular or three-dimensional (3D)-printed scaffolds are used to solve the scarcity of primary penis tissue samples. However, inconvenience and high costs limit the wide application of such methods. Here, we describe a simple, fast, and economical method of constructing artificial CC-like tissue. Human CC fibroblasts (FBs), endothelial cells (ECs), and smooth muscle cells (SMCs) were expanded in vitro and mixed with Matrigel in specific proportions. A large number of bubbles were formed in the mixture by vortexing combined with pipette blowing, creating a porous, spongy, and spatial structure. The CC FBs produced a variety of signaling factors, showed multidirectional differentiation potential, and grew in a 3D grid in Matrigel, which is necessary for CC-like tissue to maintain a porous structure as a cell scaffold. Within the CC-like tissue, ECs covered the surface of the lumen, and SMCs were located inside the trabeculae, similar to the structure of the primary CC. Various cell components remained stable for 3 days in vitro, but the EC content decreased on the 7th day. Wingless/integrated (WNT) signaling activation led to lumen atrophy and increased tissue fibrosis in CC-like tissue, inducing the same changes in characteristics as in the primary CC. This study describes a preparation method for human artificial CC-like tissue that may provide an improved experimental platform for exploring the function and structure of the CC and conducting drug screening for erectile dysfunction therapy.

Berberine Enhances Intestinal Mucosal Barrier Function by Promoting Vitamin D Receptor Activity.

OBJECTIVE: To evaluate if berberine can act on vitamin D receptors (VDR) and thereby regulate the expression of tight junction proteins (TJPs) in irritable bowel syndrame-diarrhea-predominant (IBS-D) rats. METHODS: The newborn rats were induced into IBS-D rat model via neonatal maternal separation combined with acetic acid chemical stimulation. After modeling, the model was evaluated and rats were divided into the control group and berberine treatment groups (0.85, 1.7 and 3.4 mg/kg, once a day for 2 weeks). The distal colon was obtained and colonic epithelial cells (CECs) were isolated and cultured after IBS-D model evaluation. The vitamin D receptor response element (VDRE) reporter gene was determined in the CECs of IBS-D rats to analyze the effect of berberine on the VDRE promoter. VDR overexpression or silencing technology was used to analyze whether VDR plays a role in promoting intestinal barrier repair, and to determine which region of VDR plays a role in berberine-regulated intestinal TJPs. RESULTS: The IBS-D rat model was successfully constructed and the symptoms were improved by berberine in a dose-dependent manner (P<0.05). The activity of VDRE promoter was also effectively promoted by berberine (P<0.05). Berberine increased the expression of TJPs in IBS-D CECs (P<0.05). VDR expression was significantly increased after transfection of different domains of VDR when compared to normal control and basic plasmid groups (all P<0.05). RT-qPCR and Western blot results showed that compared with the blank group, expressions of occludin and zonula occludens-1 were significantly higher in VDR containing groups (all P<0.05). Berberine plus pCMV-Myc-VDR-N group exerted the highest expression levels of occludin and zonula occludens-1 (P<0.05). CONCLUSION: Berberine enhances intestinal mucosal barrier function of IBS-D rats by promoting VDR activity, and the main site of action is the N-terminal region of VDR.

Risk Factors and Nomogram for Postoperative Pulmonary Infection in Patients with Cervical Spinal Cord Injury.

OBJECTIVE: To identify the risk factors for developing postoperative pulmonary infection in patients with acute cervical spinal cord injury (CSCI), and to develop a nomogram prediction model. METHODS: Patients with CSCI who were admitted to 3 different medical centers between July 2011 and July 2021 were included in this study. All patients underwent cervical spine surgery. Data for patients admitted to the first 2 centers were included in a training set to establish the nomogram prediction model, and data for patients admitted to the third center were included in a validation set to externally verify the efficacy of the prediction model. For the training set, patients were divided into an infected group and a noninfected group (control group). Independent risk factors for postoperative pulmonary infection in patients with CSCI were identified by univariate and multivariate logistic regression analyses. Additionally, a nomogram prediction model was developed and validated based on the risk factors. RESULTS: A total of 689 patients were enrolled, including 574 for the training set and 115 for the validation set. Of the patients included for the training set, 144 developed pulmonary infection, with an incidence of 25.09%; 40 patients included for the validation set developed pulmonary infection (34.78%). Multivariate logistic regression analysis showed that age, American Spinal Injury Association grade, steroid pulse, high-level injury, smoking, multistage surgery, and operation duration were risk factors for the development of postoperative pulmonary infection in patients with CSCI. The area under the curve of the receiver operating characteristic curve of the model built by the training set was 0.905, and that of the receiver operating characteristic curve of the verification set was 0.917. The decision curve indicated that the model was in the range 1%-100%, and the predicted net benefit value of the model was high. CONCLUSIONS: Age, American Spinal Injury Association grade, steroid pulse, CSCI site, smoking history, number of surgical levels, and surgical duration are correlated with the development of postoperative pulmonary infection in patients with CSCI. The risk prediction model of postoperative pulmonary infection has a good prediction efficiency and accuracy.

Association Between Long-Term Visit-to-Visit Hemoglobin A1c and Cardiovascular Risk in Type 2 Diabetes: The ACCORD Trial.

Background: To explore the association between visit-to-visit variability of glycated hemoglobin (HbA1c) and cardiovascular outcomes in the patients with type 2 diabetes mellitus (T2DM) of the Action to Control Cardiovascular Risk in Diabetes (ACCORD) study. Methods: We conducted a post-hoc analysis on the ACCORD population including 9,544 participants with T2DM. Visit-to-visit variability of HbA1c was defined as the individual SD, coefficient of variation (CV), and variability independent of the mean (VIM) across HbA1c measurements. The clinical measurements included primary outcome [the first occurrence of non-fatal myocardial infarction (MI), non-fatal stroke or cardiovascular death], total mortality, cardiovascular death, non-fatal MI event, non-fatal stroke, total stroke, heart failure, macrovascular events, and major coronary events (CHD). Results: Over a median follow-up of 4.85 years, 594 and 268 participants experienced all-cause mortality and cardiovascular mortality, respectively. After adjusting for baseline HbA1c levels and confounding factors, the adjusted hazard ratio (HR) comparing patients in the highest vs. the lowest quartile CV of HbA1c variability was 1.61 (95% CI 1.29-2.00) for the primary outcome. Similar trends for secondary outcome were also observed. There was no association between HbA1c fluctuation and non-fatal stroke. Noticeably, there was 66% greater risk for the all-cause mortality among patients in the highest vs. the lowest quartile (HR 1.66, 95% CI 1.27-2.17). Conclusions: Greater variability of HbA1c is associated with higher risk for cardiovascular complications and all-cause death in T2DM. Our study stresses the significance of well-controlled glycemic levels for improving cardiovascular outcomes. Further randomized clinical trials are required to confirm these findings.

Efficacy of Tong-Xie-Yao-Fang granule and its impact on whole transcriptome profiling in diarrhea-predominant irritable bowel syndrome patients: study protocol for a randomized controlled trial.

BACKGROUND: Irritable bowel syndrome (IBS) is one kind of common functional bowel disease with obscure pathogenesis, and exploration about whole transcriptome profiling in IBS-D is still negligible. Conventional medications have limited effects, which makes focus shifted to traditional Chinese medicine (TCM). Tong-Xie-Yao-Fang, as a classic herbal formula in TCM, is pretty effective and safe for the treatment of diarrhea-predominant irritable bowel syndrome (IBS-D), but the underlying therapeutic mechanism remains unknown. We aim to verify the efficacy and safety of TXYF granule (the formula particles mixed together) in IBS-D and elucidate the gene-level mechanism of IBS-D and therapeutic targets of TXYF granule based on whole transcriptome analysis. METHODS/DESIGN: This is a randomized, double-blind, and placebo-controlled clinical trial consisting of 2 weeks of run-in period, 12 weeks of treatment period, and 8 weeks of follow-up period. We will enroll 120 participants with IBS-D, who will be randomly assigned to the TXYF granule group and the placebo group, and recruit additional 10 healthy individuals as controls for mechanistic outcome. The two groups respectively take TXYF granule or placebo orally for treatment. The primary outcome is the response rate of IBS-Symptom Severity Score (IBS-SSS). The secondary outcomes include adequate relief (AR), IBS-Quality of Life Questionnaire (IBS-QOL), and long-term efficacy. Mechanistic outcome is the whole transcriptome profiling of the intestinal mucosae from IBS participants before and after the treatment and healthy individuals. DISCUSSION: This trial will prove the effectiveness and safety of TXYF granule with high-quality evidence and provide a penetrating and comprehensive perspective on the molecular mechanism of IBS-D by whole transcriptome analysis, which makes us pinpoint specific biomarkers of IBS-D and therapeutic targets of TXYF. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR-IOR-1900021785 . Registered on 9 March 2019.

(+)-Cholesten-3-one induces osteogenic differentiation of bone marrow mesenchymal stem cells by activating vitamin D receptor.

In our previous reports, it was revealed that steroids in traditional Chinese medicine (TCM) have the therapeutic potential to treat bone disease. In the present study, an in vitro model of a vitamin D receptor response element (VDRE) reporter gene assay in mesenchymal stem cells (MSCs) was used to identify steroids that enhanced osteogenic differentiation of MSCs. (+)-cholesten-3-one (CN), which possesses a ketone group that is modified in cholesterol and cholesterol myristate, effectively promoted the activity of the VDRE promoter. Phenotypic cellular analysis indicated that CN induced differentiation of MSCs into osteogenic cells and increased expression of specific osteogenesis markers, including alkaline phosphatase, collagen II and Runt-related transcription factor 2. Furthermore, CN significantly increased the expression of osteopontin, the target of the vitamin D receptor (VDR), which indicated that CN may activate vitamin D receptor signaling. Over-expression of VDR or knockdown studies with VDR-small interfering RNA revealed that the pro-differentiation effects induced by CN required VDR. Furthermore, the present study determined that the C-terminal region of the VDR is responsible for the action of CN. Taken together, the present findings demonstrated that CN induced osteogenic differentiation of MSCs by activating VDR. The present study explored the regulation of stem cells by using a series of similar steroids and provided evidence to support a potential strategy for the screening of novel drugs to treat bone disease in the future.

[Gut microbiota and osteoporosis].

OBJECTIVE: The rapidly evolving aging society in China is associated with increased incidences of osteoporosis and fractures, which have become common health problems that threaten the quality of life of the elderly. Gut microbiota colonizing in the human intestinal tract form a mutual symbiotic relationship with the host and play an important role in the metabolism and immune regulation of the host. In recent years increasing studies have demonstrated that gut microbiota not only affect the digestive system but also contribute to the disease conditions involving the immune system, and have a close relationship with the occurrence and progression of osteoporosis. This review summarizes the progress and hotspots in recent researches of the associations among gut microbiota, the immune system, osteoporosis.

A novel 3D porous metal-organic framework based on trinuclear cadmium clusters as a promising luminescent material exhibiting tunable emissions between UV and visible wavelengths.

A novel 3D porous MOF built with trinuclear cadmium clusters exhibiting rhombohedral topology has been synthesized and characterized as a promising luminescent material that can give tunable emissions between UV and visible wavelengths by controlling the number of guest molecules.

[A case-control study on risk factors for nosocomial infection of epilepsy in hospitalized children].

OBJECTIVE: To determine the incidence and risk factors for nosocomial infection in child epilepsy. METHODS: A retrospective study was conducted among all (292 cases) hospitalized children epilepsy patients in First Affiliated Hospital of Fujian Medical University from 1996 to 2000 in Fuzhou city. With all patients with nosocomial infection as cases and all patients without nosocomial infection as controls, a case-control study on risk factors for nosocomial infection was carried out. Available data were analyzed by one-way Chi-square test and unconditional logistic multiple regression model. RESULTS: One hundred fourteen cases of nosocomial infection were identified among 292 cases with epilepsy with an incidence of 39.0% (114/292). The one-way Chi-square test showed that nosocomial infection was significantly associated with age below 3 years (OR = 2.55, P < 0.01), length of hospitalization over 14 days (OR = 4.75, P < 0.01), low intelligence (OR = 3.13, P < 0.01), receiving antibiotic unreasonably (OR = 3.51, P < 0.01), using gastrogarage (OR = 3.12, P < 0.01), other invasive operation (OR = 1.85, P < 0.05) dyskinesia or palsy (OR = 3.51, P < 0.01), and urinary nitrogen beyond normal range (OR = 5.00, P < 0.05), etc. Unconditional logistic multiple regression analysis revealed that the length of hospitalization over 14 days (OR = 4.30, OR 95% CI: 2.48 - 7.46, P < 0.01), taking antibiotic unreasonably (OR = 2.74, OR 95% CI: 1.30 - 5.77, P < 0.01), using gastrogarage (OR = 3.04, OR 95% CI: 1.28 - 7.18, P < 0.05), and low intelligence (OR = 2.32, OR 95% CI: 1.34 - 4.01, P < 0.01) were independent risk factors for nosocomial infection. The tendency chi-square test showed that the longer stay in the hospital with more kinds of antibiotic used and more gastrogarage they used, the greater the risk of nosocomial infection was. CONCLUSION: Data suggested that occurrence of the nosocomial infection of children epilepsy patients was correlated with the length of hospitalization over 14 days, unreasonable using antibiotic, using gastrogarage and low intelligence.