Shuanglongjiegu pill promoted bone marrow mesenchymal stem cell osteogenic differentiation by regulating the miR-217/RUNX2 axis to activate Wnt/ß-catenin pathway.

This study aimed to investigate the effects of Shuanglongjiegu pill (SLJGP) on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and explore its mechanism based on miR-217/RUNX2 axis. Results found that drug-containing serum of SLJGP promoted BMSCs viability with a dose-dependent effect. Under osteogenic differentiation conditions, SLJGP promoted the expression of ALP, OPN, BMP2, RUNX2, and the osteogenic differentiation ability of BMSCs. In addition, SLJGP significantly reduced miR-217 expression, and miR-217 directly targeted RUNX2. After treatment with miR-217 mimic, the promoting effects of SLJGP on proliferation and osteogenic differentiation of BMSCs were significantly inhibited. MiR-217 mimic co-treated with pcDNA-RUNX2 further confirmed that the miR-217/RUNX2 axis was involved in SLJGP to promote osteogenic differentiation of BMSCs. In addition, analysis of Wnt/ß-catenin pathway protein expression showed that SLJGP activated the Wnt/ß-catenin pathway through miR-217/RUNX2. In conclusion, SLJGP promoted osteogenic differentiation of BMSCs by regulating miR-217/RUNX2 axis and activating Wnt/ß-catenin pathway.

miR5298b regulated taxol biosynthesis by acting on TcNPR3, resulting in an alleviation of the strong inhibition of the TcNPR3-TcTGA6 complex in Taxus chinensis.

Taxol, a valuable but rare secondary metabolite of the genus Taxus, is an effective anticancer drug. Understanding the regulation of taxol biosynthesis may provide a means to increase taxol content. The microRNA miR5298b was found to promote the accumulation of taxol and upregulate several taxol biosynthesis genes, including DBAT, TASY, and T5H, as demonstrated by experiments using the overexpression and mimicry of transient leaves. Moreover, miR5298b cleaves the mRNA sequence of TcNPR3, a homolog of the salicylic acid receptor AtNPR3/4. Overexpression and knockdown by RNA interference of TcNPR3 confirmed that it repressed taxol biosynthesis. These results indicate that miR5298b enhances taxol biosynthesis via the cleavage of TcNPR3. Yeast two-hybrid bimolecular fluorescence complementation and pull-down assays revealed that TcTGA6, a TGA transcription factor, physically interacted with TcNPR3. Functional experiments showed that TcTGA6 negatively regulates taxol biosynthesis by directly combining with the TGACG motif in the promoters of TASY, T5H, and T10H. TcNPR3 enhances TcTGA6 inhibition Luciferase assays showed that miR5298b alleviated the repression of the TcNPR3-TcTGA6 complex. In summary, miR5298b can cleave TcNPR3, thereby alleviating the inhibition of the TcNPR3-TcTGA6 complex to upregulate taxol biosynthesis genes.

Significance and Potential Role of Eosinophils in Non-Cystic Fibrosis Bronchiectasis.

Bronchiectasis is a complex and heterogeneous disease with a myriad of pulmonary and extrapulmonary etiologies. Bronchiectasis has a predominantly neutrophilic inflammatory profile. However, eosinophilic inflammation has also been documented in both the airways and the systemic circulation. Various diseases (eg, asthma, allergic bronchopulmonary aspergillosis, chronic rhinosinusitis with nasal polyps) characterized by heightened type 2 airway inflammatory responses, including blood or sputum eosinophilia, may coexist with bronchiectasis. Apart from those eosinophilic etiologies or comorbidities related to bronchiectasis, around 20% of patients with bronchiectasis have peripheral eosinophilia (at least 3% or 300 eosinophils/µL) with no identified concomitant disease (also termed "eosinophilic bronchiectasis"), whose roles have not been fully understood. The two key points regarding these observations are that eosinophils confer both bactericidal and antiviral properties against common pathogenic microorganisms that are usually detected in bronchiectasis, and that eosinophilic bronchiectasis has been associated with better therapeutic response to inhaled corticosteroids and other anti-TH2 profile treatments. In this review, we summarize the most significant evidence regarding the role of eosinophils in patients with bronchiectasis, including the association of bronchiectasis with eosinophilic diseases (as etiologies or comorbidities), and existing data on eosinophilic bronchiectasis not related to eosinophilic disorders.

Effects of 5-aminosalicylates or thiopurines on the progression of low-grade dysplasia in patients with inflammatory bowel disease: a systematic review and meta-analysis.

PURPOSE: Although 5-aminosalicylates and thiopurines may have an antineoplastic effect on colorectal neoplasia in patients with inflammatory bowel disease (IBD), their impact on the progression of low-grade dysplasia (LGD) in IBD is uncertain. Therefore, we performed a systematic review and meta-analysis to evaluate whether 5-aminosalicylates or thiopurines can protect against the progression of LGD in patients with IBD. METHODS: Systematic searches of PubMed, EMBASE, Cochrane Library databases, and major conference proceedings were conducted to identify all eligible studies through March 2020. Data were pooled using a random effects model. Study quality was assessed using the Newcastle-Ottawa Scale. RESULTS: Five studies comprising 776 IBD patients with LGD were included. Overall, 5-aminosalicylates (Hazard ratio (HR) = 0.91, 95% confidence interval (CI) 0.55-1.51) and thiopurines (HR = 0.64, 95% CI 0.23-1.79) did not significantly reduce the risk of advanced colorectal neoplasia (high-grade dysplasia/cancer) in IBD patients with LGD. Moreover, the effects of 5-aminosalicylates or thiopurines on risk of advanced colorectal neoplasia in IBD patients with LGD were not significant by different primary sclerosing cholangitis status, study quality, sample size, and IBD type. CONCLUSIONS: In this study, we did not find a significant protective effect of 5-aminosalicylates or thiopurines on the progression of LGD in patients with IBD.

Opportunistic Infection in Hospitalised Patients with Inflammatory Bowel Disease.

OBJECTIVE: To investigate the incidence, clinical features, and risk factors of opportunistic infections in elderly patients with inflammatory bowel disease (IBD). STUDY DESIGN: Observational study. PLACE AND DURATION OF STUDY: Department of Digestive and Geriatrics Center, Sichuan University West China Hospital, China between January 2012 and January 2019. METHODOLOGY: Patients (≥18 years) with IBD were enrolled in this study. Clinical data from the infected elderly group (age ≥60 years), non-infected elderly group (age ≥60 years) and infected adult group (age: 18-59 years) were compared. Logistic regression analysis was used for risk factors associated with opportunistic infection. RESULTS:  A total of 8.9% (307/3,456) of patients with IBD had opportunistic infection. The opportunistic infection rate of elderly group was 16.5% (80/485), which was significantly higher than that of adult group (7.6%, 227/2,971, p <0.05). Compared with infected adult group or non-infected elderly group, infected elderly group had less fever and leukocytosis, but more hypoproteinemia and several activities (p <0.05). Cytomegalovirus and Epstein-Barr virus were the most common agents in elderly group and adult group, respectively. Multiple episodes (three or more) were more common in infected elderly group; the time of opportunistic infections was associated with systemic inflammatory reaction syndrome (SIRS, p <0.05). Logistic regression analysis showed that age ≥60 years, corticosteroids, immunosuppressive and biological agents were risk factors for opportunistic infections in patients with IBD. CONCLUSION: Hospitalised elderly IBD patients, receiving corticosteroids, immunosuppressive, and biological agents, are at higher risk for infection. The symptoms of opportunistic infections in elderly patients are atypical, but they are prone to multiple infections with poor prognosis. Key Words: Elderly patients, Inflammatory bowel disease, Opportunistic infection, Systemic inflammation reaction syndrome.

Glial-derived neurotrophic factor regulates enteric mast cells and ameliorates dextran sulfate sodium-induced experimental colitis.

BACKGROUND & AIMS: Although interactions between enteric glial cells (EGCs) and enteric mast cells have been demonstrated to play an important role in the pathogenesis of inflammatory bowel disease (IBD), the exact mechanisms by which EGCs regulate enteric mast cells are still unknown. The aims of this study were to investigate whether glial-derived neurotrophic factor (GDNF), which has been confirmed to be produced mostly by EGCs, might regulate enteric mast cells and ameliorate dextran sulfate sodium (DSS)-induced experimental colitis. METHODS: Recombinant adenoviral vectors encoding GDNF (Ad-GDNF) were administered intracolonically in experimental colitis induced by DSS. The disease activity index and histological score were measured. The expression of tumour necrosis factor-α (TNF-α), interleukin-6 and myeloperoxidase (MPO) activity were measured by ELISA assay. The expression of trypsin and ß-hexosaminidase were evaluated. GDNF specific receptor (GFR-α1/RET) was detected. The calcium reflux was tested by microplate reader. The expression p-JNK was analyzed by western blot assay. RESULTS: GDNF resulted in a significant inhibition of the activation of enteric mast cells by down-regulating JNK signal pathway, lessening intracellular calcium influx, and then reducing the degranulation as well as the expression of pro-inflammatory cytokines via combing with its receptor (GFR-α1/RET) in mast cells, and these inhibitory effects were abrogated by treatment with neutralizing antibody against GDNF. Moreover, the administration of GDNF led to an amelioration of experimental colitis. CONCLUSIONS: GDNF are able to regulate enteric mast cells and ameliorate experimental colitis. GDNF might be an important mediator of the cross-talk between EGCs and enteric mast cells, and GDNF might be a useful therapeutic drug for IBD.

Gelsolin-like actin-capping protein has prognostic value and promotes tumorigenesis and epithelial-mesenchymal transition via the Hippo signaling pathway in human bladder cancer.

BACKGROUND: Transitional cell carcinoma (TCC) of the bladder, the major histologic subtype of bladder cancer, is increasing in incidence and mortality, which requires the identification of effective biomarkers. Actin-regulating proteins have recently been proposed as important antitumor druggable targets. As a gelsolin-family actin-modulating protein, CAPG (gelsolin-like actin-capping protein) generated great interest due to its crucial effects in various biological and physiological processes; however, the role and mechanism of CAPG in TCCs remain unknown. MATERIALS AND METHODS: Bioinformatic analysis and immunohistochemistry of clinical specimens were performed to detect the expression level of CAPG. Both in vitro and in vivo assays were used to determine the oncogenic effect of CAPG in TCCs. Male 4-5-week-old BALB/c nude mice were used for in vivo tumorigenesis assays, while SCID mice were used for in vivo metastatic assays. Affymetrix microarray was used to identify the underlying molecular mechanism. Western blot and immunofluorescence were used to validate the expression and localization of proteins. RESULTS: CAPG was frequently upregulated in TCCs and associated with clinical aggressiveness and worse prognosis. Functional assays demonstrated that CAPG could contribute to the tumorigenesis, metastasis and epithelial-mesenchymal transition (EMT) of TCCs both in vitro and in vivo. A novel mechanism that CAPG promoted TCC development via inactivating the Hippo pathway, leading to a nucleus translocation of Yes-associated protein was suggested. CONCLUSIONS: The current study identified CAPG as a novel and critical oncogene in TCCs, supporting the pursuit of CAPG as a potential target for TCC intervention.

MicroRNA-146a promote cell migration and invasion in human colorectal cancer via carboxypeptidase M/src-FAK pathway.

Colorectal cancer (CRC) is one of the most common cancers worldwide, and microRNAs play important roles in CRC progression. This study aimed to investigate the roles of miR-146a-5p in human CRC and their molecular mechanisms. First, we found that miR-146a-5p was significantly upregulated in CRC tissues and promoted the migration of CRC cells. Then, we identified carboxypeptidase M (CPM) as a direct target of miR-146a-5p, and found that it inhibited the migration and invasion of CRC cells. Our results also showed that CPM expression was positively correlated with overall survival and negatively correlated with recurrence, lymph node invasion, and N stage. Furthermore, we demonstrated that both miR-146a-5p and CPM regulated Src and FAK expression, while the Src-FAK signaling pathway is widely known to be associated with the migration and invasion of multiple tumor cells. This study is the first to demonstrate the functional and mechanistic relationship of the miR-146a-5p/CPM/Src-FAK axis and its effect on the migration and invasion of CRC cells. Thus, miR-146a-5p represents potential targets for CRC diagnosis and therapy.

[Identification of ubiquitously expressed transcript as the potential interactor of hepatitis B virus polymerase].

OBJECTIVE: To investigate the function of hepatitis B virus polymerase (HBV Pol) in the viral life cycle by screening the proteins interacting with HBV polymerase. METHODS: The HBV Pol gene was constructed into the pGBKT7 vector. GAL4 yeast two-hybrid system was used to screen the human liver cDNA library to obtain proteins which interacted with HBV Pol. GST-pull down assay was applied to confirm the protein interactions. RESULTS: Ubiquitously expressed transcript (UXT) was selected by the yeast two-hybrid system. GST-pull down assay confirmed the in vitro interaction between HBV Pol and UXT. CONCLUSIONS: UXT is a potential interactor of HBV Pol, and this protein interaction may provide clues of the function of HBV Pol in HBV life cycle.

[The relationship between PTEN expression and PI3K/AKT signal pathway in colon carcinoma].

OBJECTIVE: To investigate the expression of PTEN in carcinogenesis and development of colon carcinoma and to reveal their correlation with the PI3K/AKT pathway. METHODS: The expressions of PTEN and p-AKT were detected by immunohistochemistry and western blotting in 32 specimens from patients with colon carcinoma. RESULTS: The positive rate of p-PTEN was significantly lower in colon carcinoma (31.25%) than that in normal control tissues (96.87%, P<0.01), and the positive rate of p-AKT was significantly higher in colon carcinoma (87.5%) than that in normal control tissues (15.62%, P<0.01), and expression level of PTEN and p-AKT were related to histological differentiation, distant invasion, lymph node metastasis and Duke's stage (P<0.05). A negative correlation was observed between the expression of PTEN and p-AKT proteins. (r=-0.872, P<0.01). CONCLUSION: The over-expression of AKT accompanied by the loss of PTEN involves carcinogenesis and development of colon carcinoma.

[Effects of aging on the expressions of insulin receptor substrate-1, phosphate protein kinase B in rat liver and muscle].

OBJECTIVES: To investigate the effects of aging on the levels of free fatty acid (FFA), lipid profile, the expression of IRS-1 and its downstream signal Akt phosphorylation, so to explore the mechanisms of aging-associated glucose tolerance impairment. METHODS: Three different age groups (6, 12, 20-24 months) of SD rats were used. After being fasted for 12-14 h, the rats were anesthetized with sodium pentobarbital (30 mg/kg BW, i.p.), then the samples of blood, liver and muscle were collected. Blood samples were used for the detection of fasting blood glucose, lipid profile and FFA. The expressions of IRS-1 and p-Akt in liver and muscle were measured by immunohistochemical staining method. RESULTS: 1) The level of FFA in 20-24 months groups was significantly higher [(419.94 +/- 93.93) vs (256.22 +/- 73.93) mmol/L, P<0.05] than that of younger group (6 months); 2) IRS-1 was expressed in liver and muscle cell of all groups. The level of IRS-1 showed no significant change in both liver and muscle. 3) p-Akt expression in liver significantly decreased in old rat when comparing to younger counterparts [2911.06 +/- 268.13 vs 4683.72 +/- 582.29 (12 months) & 4903.06 +/- 688.44 (6 months), P<0.05], while no difference was found in muscle among three age groups. CONCLUSION: 1) FFA was the initial metabolic change in natural aging SD rat, which might play an important role in age-related insulin resistance. 2) There was no significant difference of IRS-1 expression among three rat age groups. 3) PI3K/Akt pathway in liver of old rat is a critical signal defect of insulin dysfunction with aging. Liver might be main component organ involved to age-related insulin resistance.

[Long-term follow-up results of synovectomy for rheumatoid knee].

OBJECTIVE: To continuously observe the long-term effects of synovectomy for improving joint damage and quality-of-life in patients with the rheumatoid knee. METHODS: Twenty-one consecutive patients with rheumatoid arthritis (RA) involving 24 knees underwent open synovectomy from November 1988 to January 1997 between November 1988 and January 1997. The changes in radiographic damage were assessed with Larsen score on plain films before and 6 months after surgery with subsequent annual assessment for 8 years, and the functional recovery of the patients was also evaluated with Health Assessment Questionnaire (HAQ) at the same time. RESULTS: The radiographic joint damage and juxta-articular osteoporosis or bone erosion was ameliorated after surgery in all the patients. Larsen score began to decrease 6 months after the operation, and the best effects were achieved at one year and maintained for at least 5 years after the operation, but then followed by recurrence of joint lesions. HAQ scores were improved after the surgery with the best effects observed 6 months after the operation lasting for over 2 years. HAQ score gradually decreased 4 years after the operation till reaching the preoperative scores. CONCLUSION: Synovectomy in the patients with rheumatoid knee not only reverses progressive joint damage, but also improves juxta-articular bone erosions and the patients' quality of life. However, radiographic joint damage and functional deterioration may recur due to hyperplasia of the inflammatory synovium in the long term after operation, suggesting that the inflammatory synovium participates in local joint damage with bone erosions and systemic pathologic process of RA.