MicroRNA-21: An Emerging Player in Bone Diseases.

MicroRNAs (MiRNAs) are small endogenous non-coding RNAs that bind to the 3'-untranslated region of target genes and promote their degradation or inhibit translation, thereby regulating gene expression. MiRNAs are ubiquitous in biology and are involved in many biological processes, playing an important role in a variety of physiological and pathological processes. MiRNA-21 (miR-21) is one of them. In recent years, miR-21 has received a lot of attention from researchers as an emerging player in orthopedic diseases. MiR-21 is closely associated with the occurrence, development, treatment, and prevention of orthopedic diseases through a variety of mechanisms. This review summarizes its effects on osteoblasts, osteoclasts and their relationship with osteoporosis, fracture, osteoarthritis (OA), osteonecrosis, providing a new way of thinking for the diagnosis, treatment and prevention of these bone diseases.

Photothermal Effect-based Cytotoxic Ability of Melanin from Mytilus edulis Shells to Heal Wounds Infected with Drug-resistant Bacteria in vivo.

OBJECTIVE: Owing to antibiotic abuse and the subsequent development of antibiotic resistance, bacterial infection has become one of the most persistent unresolved problems. New antibacterial agents, especially those that are environmental-friendly, are urgently needed. METHODS: Melanin extracted by filtration centrifugation and acid and proteolytic hydrolysis was characterized using UV, FTIR, TEM, and XPS. Photothermal conversion was calculated, and the bacteriostatic effects, in vitro and in vivo, were assessed by plate counting and ratios (%) of wound areas. RESULTS: Natural melanin hydrolyzed by trypsin had good photothermal conversion effects, which resulted in superior bacteriostatic activities. The extracted melanin along with laser NIR irradiation at 808 nm promoted the healing of wounds infected by drug-resistant bacteria in vivo and was biocompatible according to toxicity tests in vivo and in vitro. CONCLUSION: The present findings indicated a safe and efficient method of developing natural antibacterial agents.

Catharmus tinctorius volatile oil promote the migration of mesenchymal stem cells via ROCK2/Myosin light chain signaling.

MSC transplantation has been explored as a new clinical approach to stem cell-based therapies for bone diseases in regenerative medicine due to their osteogenic capability. However, only a small population of implanted MSC could successfully reach the injured areas. Therefore, enhancing MSC migration could be a beneficial strategy to improve the therapeutic potential of cell transplantation. Catharmus tinctorius volatile oil (CTVO) was found to facilitate MSC migration. Further exploration of the underlying molecular mechanism participating in the pro-migratory ability may provide a novel strategy to improve MSC transplantation efficacy. This study indicated that CTVO promotes MSC migration through enhancing ROCK2 mRNA and protein expressions. MSC migration induced by CTVO was blunted by ROCK2 inhibitor, which also decreased myosin light chain (MLC) phosphorylation. Meanwhile, the siRNA for ROCK2 inhibited the effect of CTVO on MSC migration ability and attenuated MLC phosphorylation, suggesting that CTVO may promote BMSC migration via the ROCK2/MLC signaling. Taken together, this study indicates that C. tinctorius volatile oil could enhance MSC migration via ROCK2/MLC signaling in vitro. C. tinctorius volatile oil-targeted therapy could be a beneficial strategy to improve the therapeutic potential of cell transplantation for bone diseases in regenerative medicine.

Quantitative detection method of Enterocytozoon hepatopenaei using TaqMan probe real-time PCR.

A TaqMan probe and a pair of specific primers were selected from the small subunit ribosomal DNA (SSU rDNA) sequence of Enterocytozoon hepatopenaei (EHP); this real-time PCR assay was developed and optimized. It showed a good linearity in detecting standards of EHP SSU rDNA fragments from 4 × 102 to 4 × 108 copies/reaction using the established method. The detection limit of the qPCR method was as low as 4 × 101 copies per reaction, which was higher than the conventional PCR and SYBR Green I-based EHP qPCR reported. Using the qPCR assay, EHP was detected in four batches of slow-growing Penaeus vannamei specimens collected from Tianjin and Zhejiang Province in China was detected using qPCR. The results showed that all the hepatopancreas from the slow-growing P. vannamei specimens were detected as EHP-positive. EHP copies of hepatopancreas in some batches had a negative correlation with the body mass index (BMI) of shrimps; however, not all batches of specimens had this negative correlation between EHP copies of hepatopancreas and BMI. This qPCR technique is sensitive, specific and easy to perform (96 tests in <3 h), which provides technical support for the detection and prevention of EHP.

[Summarize drug dosage forms in treatment of age-related macular degeneration disease].

In this review, the authors summarized the drugs in treatment of the age-related macular degeneration (AMD or ARMD), including the pathogenesis of the age-related macular degeneration at home and abroad, dosage forms used in the treatment, and the drugs research and development directions in the future. AMD disease is the third largest blinding diseases all over the world, with an incidence of 6.62%. The dosage form of the traditional medicine is mostly oral formulations, playing a role in body, while the newly dosage form is topical drug delivery formulation. Traditional Chinese medicine (TCM) has certain advantages in the treatment of AMD disease and the development of topical drug delivery preparations with newly preparation technologies would have a very bright prospect in the future.

(+)-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.

Anti-inflammatory effect of the blueberry anthocyanins malvidin-3-glucoside and malvidin-3-galactoside in endothelial cells.

Blueberry fruits have a wide range of health benefits because of their abundant anthocyanins, which are natural antioxidants. The purpose of this study was to investigate the inhibitory effect of blueberry's two main anthocyanins (malvidin-3-glucoside and malvidin-3-galactoside) on inflammatory response in endothelial cells. These two malvidin glycosides could inhibit tumor necrosis factor-alpha (TNF-α) induced increases of monocyte chemotactic protein-1 (MCP-1), intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) production both in the protein and mRNA levels in a concentration-dependent manner. Mv-3-glc at the concentration of 1 µM could inhibit 35.9% increased MCP-1, 54.4% ICAM-1, and 44.7% VCAM-1 protein in supernatant, as well as 9.88% MCP-1 and 48.6% ICAM-1 mRNA expression (p<0.05). In addition, they could decrease IκBα degradation (Mv-3-glc, Mv-3-gal, and their mixture at the concentration of 50 µM had the inhibition rate of 84.8%, 75.3%, and 43.2%, respectively, p<0.01) and block the nuclear translocation of p65, which suggested their anti-inflammation mechanism was mediated by the nuclear factor-kappa B (NF-κB) pathway. In general malvidin-3-glucoside had better anti-inflammatory effect than malvidin-3-galactoside. These results indicated that blueberry is good resource of anti-inflammatory anthocyanins, which can be promising molecules for the development of nutraceuticals to prevent chronic inflammation in many diseases.

Inhibitory effect of Malvidin on TNF-α-induced inflammatory response in endothelial cells.

Vascular inflammatory responses are key mediators of endothelial dysfunction that leads to various pathologies in many diseases including atherosclerosis and cancer. The purpose of the study was to investigate the effects and molecular mechanisms of Malvidin, a natural pigment with strong antioxidant activity, on regulating inflammatory response in endothelial cells. Our results showed that tumor necrosis factor-alpha (TNF-α) significantly increased the protein or mRNA levels of monocyte chemotactic protein-1 (MCP-1), intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1), whereas pretreatment with Malvidin inhibited TNF-α-induced increases of MCP-1, ICAM-1, and VCAM-1 production in a concentration-dependent manner. In addition, Malvidin could inhibit degradation of IκBα and the nuclear translocation of p65, which suggesting the anti-inflammation mechanism of Malvidin by the nuclear factor kappa B (NF-κB) pathway. These results indicate the potential role of Malvidin in preventing chronic inflammation in many diseases.

Bis(3,5-di-tert-butyl-4H-1,2,4-triazol-4-amine-κN(1))(nitrato-κO)silver(I) ethanol monosolvate monohydrate.

The Ag(I) atom in the title compound, [Ag(NO(3))(C(10)H(20)N(4))(2)]·C(2)H(5)OH·H(2)O, is coordinated by the N atoms of two N-heterocycles [N-Ag-N = 151.5 (1)°]; the approximately linear coordination geometry is distorted into a T-shaped geometry owing to a long Ag⋯O(nitrate) bond [2.717 (4) Å]. The N atoms of the N-heterocycles that are not involved in coordination point towards the lattice water mol-ecule, which functions as a hydrogen-bond donor. The water mol-ecule itself is a hydrogen-bond acceptor towards the ethanol solvent mol-ecule. Hydrogen bonds of the type N-H⋯O give rise to a layer motif parallel to (001).

[Effect of shugan jianpi bushen recipe on splenic T lymphocytes and virus load in hepatitis B virus transgenic mice].

OBJECTIVE: To observe the effect of Shugan Jianpi Bushen Recipe (SJBR) on the splenic T lymphocytes and virus load in the hepatitis B virus (HBV) transgenic (Tg) mice, and to study its antiviral efficacy and mechanisms of action. METHODS: Sixty male BALB/C mice of SPF grade were included. Ten non-HBV Tg male mice were included as the normal control group. Fifty HBV Tg mice were randomly divided into five groups, i. e., the model group, the adefovir (ADV) group, the low dose SJBR group, the middle dose SJBR group, and the high dose SJBR group, ten in each. 10, 20, and 40 g/kg SJBR crude drug was respectively given by gastro-gavage to mice in the low dose SJBR group, the middle dose SJBR group, and the high dose SJBR group. ADV 50 mg/kg body weight was given by gastrogavage to mice in the ADV group. Equal volume of sterilized iso-osmia was given to mice in the normal group and the model group. The medication was performed once daily, totally for 21 successive days. The serum HBV DNA titers of HBV Tg mice were detected using Real-time fluorescent PCR one day before administration (T0), ten days after administration (T1), 21 days after administration (T2), and three days after withdrawal (T3), respectively; the serum hepatitis B surface antigen (HBsAg) of HBV Tg mice on T3 was detected by ELISA. The splenic T lymphocyte percent of all mice was detected by flow cytometry. RESULTS: Serum HBsAg at TO was positive in the high-, middle-, low-dose SJBR, and ADV groups. The HBsAg negative rate at T3 was lower in the high dose SJBR group than in the ADV group, showing statistical difference (P<0.01). Compared with TO of the same group, the serum HBV DNA titers could be continually decreased by high dose SJBR, showing statistical difference (P<0.01). The serum HBV DNA titers also gradually decreased in the ADV group (P<0.01), but it somewhat increased at T3. The CD3+ cell percent could be elevated by high-, middle-, low-dose SJBR, and ADV groups (P<0.05, P<0.01). The CD8+ T cell percent could also be obviously lowered by high-, middle-, and low-dose SJBR (P<0.01). Compared with the middle-, low-dose SJBR, and ADV groups, the CD4+ T cell percent and CD4+/CD8+ increased as well as CD8+ decreased in the high dose SJBR group, showing statistical difference (P<0.01). The CD3+ T cell percent was significantly positively correlated to the decrement of HBV DNA titers between the pre-treatment and post-treatment in the middle dose SJBR group (r=0.654, P<0.05). The percents of CD4+, CD8+ T cells and CD4+/CD8+ were significantly positively correlated to the decrement of HBV DNA titers between the pre-treatment and post-treatment in the high dose SJBR group (r=0.53, r=0.79, r =0.80, P<0.01). CONCLUSIONS: SJBR were capable of inhibiting the HBV DNA duplication of HBV Tg mice. One of its anti-HBV mechanisms possibly be improving the the abnormality of T lymphocyte subsets and the immune function.