[Effect of Taohong Siwu Decoction() early intervention on mesenchymal stem cells homing in fracture healing in rats].

OBJECTIVE: To observe the effects of Taohong Siwu Decoction(, THSWD) on the mesenchymal stem cells(MSCs) migration, homing number and cytokine expression in callus during the early process of fracture healing, and to explore the mechanism of THSWD on accelerationg fracture healing by regulating the homing of MSCs in rats. METHODS: A rat model of right femoral shaft open fracture was established. Thirty-two 5-week-old male Sprague-Dawley rats, weighting 110 to 130 g, were divided into control group, low-dose group, medium-dose group and high-dose group by using random number table. Distilled water was given to the control group, and the other groups were given Taohong Siwu Decoction. The rats were gavaged twice a day for 5 consecutive days after surgery. Bone volume/tissue volume(BV/TV) and bone mineral density(BMD) were observed using micro-computed tomography (micro-CT) at 21 days after surgery. At 5 days post-fracture, peripheral blood MSCs from THSWD treated and untreated rats were cultured in vitro. Subsequently, the migration ability of MSCs was observed by cell migration assay. The number of MSCs homing to the callus at the early stage of fracture (5 d) was detected by Immunohistochemistry (IHC). Protein chip was used to detect the expression of cytokines in callus. RESULTS: Micro-CT results showed that BV/TV was higher in the high-dose group than in the medium-dose group (P=0.032), and higher in the medium-dose group than in the low-dose group(P=0.041), with no difference between the control and low-dose group (P=0.651). In addition, there was no difference in BMD between low-dose group and the model group (P=0.671), and lower in the low-dose group than in the medium-dose group(P=0.018), and the medium-dose group was lower than the high-dose group(P=0.008). Cell migration assay showed that THSWD promotes enhanced the migration ability of peripheral blood MSCs. IHC assay revealed that CD45-, CD90+, CD29+ MSCs significantly increased in bone callus after THSWD intervention compared with the control group. Protein chip showed that THSWD promoted the upregulation of CINC-1(×2.91), CINC-3(×1.59), LIX(×1.5), Thymus Chemokine (×2.55), VEGF (×1.22) and the down-regulation of TIMP-1 (×2.98). CONCLUSION: THSWD, a representative formula of "promoting blood circulation and removing blood stasis", can significantly accelerate fracture healing, and its mechanism may be related to enhancing the migration ability of peripheral blood MSCs and up-regulating CINC-1, CINC-3, LIX, Thymus Chemokine, VEGF and down-regulating TIMP-1 in bone callus, which promotes the peripheral blood MSCs homing in the early stage of fracture.

Huanglian Jiedutang Alleviates Liver Injury in Septic Mice by Inducing Autophagy / 中国实验方剂学杂志

ObjectiveTo explore effect of Huanglian Jiedutang （HLJDT） on autophagy-related protein expression in septic mice with liver injury induced by cecal ligation and puncture （CLP）. MethodSixty eight-week-old C57BL/6 mice were randomly divided into four groups， namely， the sham operation group， model group， and low- （1.44 g∙kg-1） and high-dose （2.88 g∙kg-1） HLJDT groups， with 15 in each group. The septic model was established by CLP after the last administration of HLJDT for three successive days. The survival rate of mice with 24 h was observed. The mice were sacrificed 12 h after operation for collecting the serum and liver tissue. The levels of serum interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， and interleukin-1β （IL-1β） were detected by enzyme-linked immunosorbent assay （ELISA）， and the levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum by biochemical method. The pathological changes in liver tissue were observed by hematoxylin-eosin （HE） staining， and the apoptosis index （AI） of hepatocytes by TdT-mediated dUTP-biotin nick end-labeling （TUNEL）. The expression levels of protein high-mobility group box 1 protein （HMGB1）， Beclin1， and microtubule-associated protein 1 light chain 3 （LC3）-Ⅱ/Ⅰ in the liver tissue were assayed by Western blot. ResultCompared with the sham operation group， the model group showed reduced survival rate at 12 and 24 h， elevated IL-6， TNF-α， and IL-1β levels， enhanced AST and ALT activities （P<0.05）， hepatocyte swelling， inflammatory cell infiltration， and apoptosis， and up-regulated HMGB1 （P<0.05）， Beclin1， and LC3-Ⅱ/Ⅰ （P<0.05）. Compared with the model group， each medication group exhibited increased survival rate at 12 and 24 h， lowered IL-6 and TNF-α levels， weakened AST and ALT activities （P<0.05）， alleviated liver injury and apoptosis （P<0.05）， down-regulated HMGB1 expression （ P<0.05）， and up-regulated Beclin1 and LC3-Ⅱ/Ⅰ （P<0.05）. ConclusionHLJDT alleviates the liver injury of septic mice possibly by inducing autophagy and inhibiting apoptosis.

Tanshinone IIA Ameliorates Inflammation Response in Osteoarthritis via Inhibition of miR-155/FOXO3 Axis.

BACKGROUND: Osteoarthritis (OA) is the most common joint disorder characterized by degeneration of the articular cartilage and joint destruction with an associated risk of mobility disability in elderly people. Although a lot of achievements have been made, OA is still regarded as an incurable disease. Therefore, the pathological mechanisms and novel therapeutic strategies of OA need more investigation. METHODS: MTT assay was conducted to measure the viability of chondrocytes after LPS treatment. Cell apoptosis was analyzed by annexin V/propidium iodide labeling. ELISA was used to determine the concentrations of interleukin (IL)-1ß, IL-6, and tumor necrosis factor (TNF)-α in the culture supernatant of chondrocytes. The expression level of miR-155, IL-1ß, FOXO3, TNF-α, IL-6, caspase-3, and caspase-9 in chondrocytes was analyzed by RT-qPCR or Western blot. RESULTS: We found that LPS led to inflammatory responses, cell apoptosis, and increased miR-155 expression in human articular chondrocytes. Tanshinone IIA could inhibit LPS-induced inflammation and cell apoptosis of chondrocytes via regulating the expression of miR-155 and FOXO3. miR-155 directly targeted the 3'-UTR of FOXO3 to regulate its expression. CONCLUSIONS: Taken together, our data suggest tanshinone IIA ameliorates inflammation response in OA via inhibition of the miR-155/FOXO3 axis, and provide some evidences that tanshinone IIA could be designed and developed as a new promising clinical therapeutic drug for OA patients.

[Study on MSCs homing and its research on osteodiseases].

Mesenchymal stem cell (MSCs) has recently emerged as an appealing and potential therapeutic strategy to cure a diverse range of diseases in the orthopaedic field. Owing to its capacity of osteogenic differentiation, most of researches just focused on promoting MSC differentiation. With the in-depth study, MSCs homing is also a key issue for bone formation and bone diseases treatment, which have been described that MSCs mobilize from in situ environment (bone marrow) and migrate into injured tissues during the healing process through peripheral circulation. MSC homing is the incipient step of bone formation. MSCs need to firstly migrate to the bone surface and then differentiate into osteogenic cells to enhance bone repair. Promoting MSCs homing have been shown to improve recovery of several orthopedic diseases, such as osteoporosis, fracture, bone defect and wear-particle-related osteolysis. Therefore, further research on MSCs homing may provide a new thinking for treatment of osteoporosis.

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.

Study of wide-spectrum and high-resolution diffraction optical elements by stacks of binary phase gratings.

This work theoretically investigates wide-spectrum and high-resolution diffraction optical elements that are made of stacks of low-resolution binary phase gratings, whereby the two-dimensional grids in different grating layers are arranged with specified displacements. We remodel the common kinoform algorithm for this multi-scale architecture. Numerical computations show that, by increasing the number of stacking layers, the resolution of the far-field image can be improved and also that the optical elements are more insensitive to variations of incident wavelengths at the cost of part accuracy of the image reconstructions. Practical concern focuses on largely increasing the number of grating layers and efficiency of the optical designs in theory and on the manufacture of stacks of ultra-thin grating films.

(+)-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.

Non-invasive biopsy of doped-ions inside optical substrate by modified two-photon microscopy.

Doped-ion based optical elements play key roles in optical signal processes, including amplification, absorption, wavelength-filtering, lighting, and polarizing plate. Non-invasively mapping the spatial distribution of the ion concentrations in these optical elements is highly desirable either during the fabrication process or to determine their optical qualities. In this work, we applied modified two-photon fluorescence (m-TPF) microscopy to trace the ion-distributions deep inside the optical elements. For demonstration purposes, polyvinyl alcohol (PVA) polymer films inside polarizing plates are taken as an example, where the spatial distributions of Iodine-dyed ions were measured by the m-TPF microscope in a fast and non-invasive way. The durability of the polarizer films can be distinguished from the axial distribution of the Iodine-dyed ions, without the need to perform a biopsy. This proposed method and demonstrated results show great potential for monitoring the spatial distributions of doped-ions in the optical elements quickly and non-destructively, which would be of great benefit in both scientific research and industrial applications.

Approximate scheme by the coupled-wave theory to efficiently analyze the influences of moiré phenomena in liquid-crystal devices.

This work studies an approximate scheme by coupled-wave theory to analyze quickly the large-scale moiré phenomena as seen in common liquid-crystal devices. The moiré phenomena are considered to be caused by two periodic structures (with lattice vectors γ[combininb arrow](1) and γ[combininb arrow](2) and show an interference pattern spanning over a length γ(m)=|γ[combininb arrow](1)|·|γ[combininb arrow](2)|/|γ[combininb arrow](1)-γ[combininb arrow](2)| (with γ[combininb arrow](1)=/~γ[combininb arrow](2)). With the coupled-wave theory, the complete analysis of the moiré optics includes at least 2γ(m)/λ (λ: wavelength in vacuum) Fourier components and presents an ineffective computation. This work applies a cos(τ) type approximation for the openings of unpatterned liquid-crystal pixels, and considers the first-order coupling between the Fourier components of pixels and other (periodic) optical structures. We hence arrive at an effective evaluation, including 4τ|γ[combininb arrow](1)|/λ (or 4τ|γ[combininb arrow](2)|/λ) Fourier components, and are able to go back to a complete analysis when considering higher-order couplings at an appropriate τ integer value.

Microstructural and microspectral characterization of a vertically aligned liquid crystal display panel.

The microstructural and microspectral characteristics of a vertically aligned liquid crystal display (VA-LCD) panel were obtained noninvasively for the first time. With 1 µm axial and 2 µm transversal resolutions, the cell gap profile beneath the patterned thin-film transistor of the VA-LCD panel can clearly be resolved. The thicknesses of the multiple thin-film layers and the embedded defects can also be unveiled. As far as spectral response is concerned, the light transmittance at the layer boundaries can be estimated from the measured reflectance, which is crucial information for the design of a highly transmissive panel. The color shift of the VA-LCD panel due to fabrication error was evaluated.

Extraordinarily wide-view circular polarizers for liquid crystal displays.

A new broadband wide-view circular polarizer is proposed for high transmittance multi-domain vertical-alignment liquid crystal displays (MVA LCDs). This configuration only requires one biaxial plate in the conventional circular polarizer. The optimal film parameters are obtained analytically through spherical trigonometric method on the Poincaré sphere and through computer-aided parameter search method. According to this design, the high transmittance MVA LCD exhibits a contrast ratio CR 200:1 over approximately 80 degrees viewing cone.