Potential Anti-Inflammatory Components of Rhododendron molle G. Don Leaf Extracts in LPS-Induced RAW 264.7.

As a traditional Chinese medicine, Rhododendron molle G. Don has a long history of treating rheumatoid arthritis. In this study, RAW 264.7 cells induced by lipopolysaccharide (LPS) were established as cell inflammatory model to evaluate the anti-inflammatory activity of chloroform extract from R. molle leaves (CERL), ethyl acetate extract from R. molle leaves (EERL) and butanol extract from R. molle leaves (BERL) and analyze the potential anti-inflammatory components of R. molle. Potential anti-inflammatory components analysis of CERL were performed by HPLC and UHPLC-Q-TOF-MS. Prediction of potential anti-inflammatory components by molecular docking experiments. Compared with negative control group, 25 µg/mL CERL could reduce the release level of NO by 62 %, and the mRNA expression levels of COX-2, IL-6, IL-1ß and TNF-α were reduced by 69.74 %, 86.25 %, 77.94 % and 56.80 %, respectively. Western-Blot showed similar results. CERL, EERL and BERL exerted their inhibitory activity in dose-dependent manner. All results showed that the higher the concentration, the better the anti-inflammatory activity. CERL showed the best inhibitory activity, the second was EERL, and then was BERL. 21 terpenoids and 4 flavonoids were identified in CERL by UHPLC-Q-TOF-MS. Molecular docking results showed that triterpenoids in CERL had better interaction with target proteins (TNF-α, IL-1ß). It indicated that triterpenoids may be potential anti-inflammatory components of R. molle leaves. This study explored the anti-inflammatory activities of CERL, EERL, BERL, which laid a foundation for further promoting the clinical application of R. molle.

Anti-rheumatoid arthritis potential of Rhododendron molle G. Don leaf extract in adjuvant induced arthritis rats.

AIM OF THE STUDY: The aim of this study was to test the anti-rheumatic arthritis effects of Rhododendron molle G. Don leaf extract in arthritis rats and inflammatory RAW 264.7 cells. Preliminary analysis and comparison of potential medicinal components of three polar extracts by HPLC and UHPLC-Q-TOF-MS. MATERIALS AND METHODS: SD rats were subcutaneously injected with complete Freund's adjuvant (CFA) to induce inflammation on the right hind paw. RAW 264.7 cells were induced by lipopolysaccharide (LPS) to established cell inflammatory model. The volume of rat hind paw was measured with a volume meter to detect swelling, and the weight of rats was measured with an electronic balance. The severity of arthritis in rats was evaluated by arthritis score. The pathological sections of rat hind paw joints were observed by hematoxylin-eosin staining, and the contents of IL-6 and IL-1ß in serum were detected. qRT-PCR was used to detect the expression of IL-1ß, IL-6, TNF-α and COX-2 genes in RAW 264.7 cells. The release of nitric oxide was measured by Griess reaction. The expression levels of IL-6 and IL-1ß were detected by Western-Blot. RESULTS: and discussion: The chloroform extract from R. molle leaves (CERL), Ethyl acetate extract from R. molle leaves (EERL), n-butanol extract from R. molle leaves (BERL) could significantly inhibit hind paws swelling and reduce arthritis index in arthritis rats. And it showed dose dependence. Compared with tripterygium glycosides (TG) tablets, an effective drug of RA treatment, CERL have better anti-RA effect after administration. In addition, the three kinds of the polar extracts of Rhododendron molle leaves (PERL) had lower toxicity, with the LD50 279.87, 239.65, 500.08 (mg/kg) respectively, while TG group's LD50 was 96.00 (mg/kg). In vitro experiments showed that the three PERLs can significantly inhibit the level of pro-inflammatory factors and inflammatory mediator, such as TNF-α, IL-1ß, IL-6, COX-2 and NO, which were consistent with their anti-RA ability. Among the three kinds of PERLs, CERL showed the best inhibitory activity. CONCLUSION: The R. molle leaf is a potential medicinal part for the treatment of RA. This study explored the anti-RA and anti-inflammatory activities of CERL, EERL, BERL, which laid a foundation for further promoting the clinical application of R. molle.

State Estimation for Discrete Time-Delayed Impulsive Neural Networks Under Communication Constraints: A Delay-Range-Dependent Approach.

In this article, a delay-range-dependent approach is put forward to tackle the state estimation problem for delayed impulsive neural networks. A new type of nonlinear function, which is more general than the normal sigmoid function and functions constrained by the Lipschitz condition, is adopted as the neuron activation function. To effectively alleviate data collisions and save energy, the round-robin protocol is utilized to mitigate the occurrence of unnecessary network congestion in communication channels from sensors to the estimator. With the aid of the Lyapunov stability theory, a state observer is constructed such that the estimation error dynamics are asymptotically stable. The observer existence is ensured by resorting to a set of delay-range-dependent criteria which is dependent on both the impulsive time instant and a coefficient matrix. In addition, the synthesis of the observer is discussed by using linear matrix inequalities. Simulations are provided to illustrate the reasonability of our delay-range-dependent estimation approach.

H∞ State Estimation for BAM Neural Networks With Binary Mode Switching and Distributed Leakage Delays Under Periodic Scheduling Protocol.

This article is concerned with the H∞ state estimation problem for a class of bidirectional associative memory (BAM) neural networks with binary mode switching, where the distributed delays are included in the leakage terms. A couple of stochastic variables taking values of 1 or 0 are introduced to characterize the switching behavior between the redundant models of the BAM neural network, and a general type of neuron activation function (i.e., the sector-bounded nonlinearity) is considered. In order to prevent the data transmissions from collisions, a periodic scheduling protocol (i.e., round-robin protocol) is adopted to orchestrate the transmission order of sensors. The purpose of this work is to develop a full-order estimator such that the error dynamics of the state estimation is exponentially mean-square stable and the H∞ performance requirement of the output estimation error is also achieved. Sufficient conditions are established to ensure the existence of the required estimator by constructing a mode-dependent Lyapunov-Krasovskii functional. Then, the desired estimator parameters are obtained by solving a set of matrix inequalities. Finally, a numerical example is provided to show the effectiveness of the proposed estimator design method.

Rhododendron molle G. Don Extract Induces Apoptosis and Inhibits Migration in Human Colorectal Cancer Cells and Potential Anticancer Components Analysis.

Rhododendron molle G. Don is one example of traditional Chinese medicine with important medicinal value. In this study, the effects of methanol extract of R. molle leaves (RLE) on colorectal cancer HT-29 cells and its potential molecular mechanism were investigated. MTT analysis showed that RLE could significantly inhibit the cell viability and migration of HT-29 cells in a concentration-dependent manner. Cell cycle analyses via flow cytometer suggested that RLE induced DNA fragmentation, indicative of apoptosis, and arrest at the S phase in HT-29 cells. Quantitative real-time PCR (qRT-PCR) analysis showed that RLE could upregulate the mRNA expression of p53 and p21 in HT-29 cells, which would result in HT-29 cells being blocked in S phase. Meanwhile, RLE could upregulate the expression of Bax, and downregulate the expression of Bcl-2, which would induce cell apoptosis. Further western blot analysis showed that the protein expression changes of Bax and P53 were basically consistent with the results of qRT-PCR. In addition, GC-MS analysis detected 17 potential anticancer components in R. molle. These results indicate that R. molle has significant anticancer activity, which provides some useful information for further study and clinical application for R. molle.

Injection route affects intra-articular hyaluronic acid distribution and clinical outcome in viscosupplementation treatment for knee osteoarthritis: a combined cadaver study and randomized clinical trial.

The coverage of hyaluronic acid (HA) on the impaired cartilage should be the precondition to exert its beneficial effect on knee osteoarthritis (KOA) according to the pharmacological mechanism. However, the intra-articular distribution of HA might be correlated with the route of drug delivery. Forty-two cadaver knees with radiographic evidence of osteoarthritis were given anteromedial (AM) or medial midpatellar (MMP) injection of HA (molecular weight 600-1500 kD) followed by gait stimulation. Although 2.5 ml HA delivered through both routes failed to cover the entire cartilage, HA covered 96.12% cartilage of patellofemoral joint (PFJ) and 71.44% of medial femorotibial joint (FTJ) through MMP route, whereas mainly distributed into FTJ and posterior condyles through AM route. HA in the MMP group distributed more in PFJ than that in the AM group (P < 0.001), but no significant difference presented in medial FTJ (P = 0.084). The clinical efficacy was also associated with the route of drug delivery. One hundred patients with unilateral mild-to-moderate KOA were recruited and randomly assigned to receive five weekly HA injections with AM route (n = 50) or MMP route (n = 50). Patients in the MMP group obtained better improvement in WOMAC index total score, pain score, stiffness score, and Lequesne index total score over the entire follow-up period, as compared to patients in the AM group (all P < 0.01). More patients in the MMP group claimed pain relief (71.7%, P = 0.024) and felt satisfying (63.1%, P = 0.007) than in the AM group at the end of follow-up. Therefore, intra-articular HA injection through MMP route is recommended in treating mild-to-moderate KOA. Graphical Abstract .

[Hyaluronate acid for treatment of chondromalacia patellae: a 52-week follow-up study].

OBJECTIVE: To assess the therapeutic effect of hyaluronate acid (HA) injection through the subpatellar route for treatment of chondromalacia patellae (CP). METHODS: Eighty-eight patients with the diagnosis of CP were enrolled in this prospective study, including 38 with early CP (CP group) and 50 with advanced CP (patellofemoral arthritis group) diagnosed based on image presentations. All the patients received intra-articular HA injections through a subpatellar route once a week for 5 consecutive weeks. The primary outcome measures included WOMAC index scores and Lequesne scores before and at 4, 12, 26 and 52 weeks after the injections. The secondary outcome measures included the 30-m walking time and stair ascending and descending time (one floor) before and at 1, 2, 3, and 4 weeks after the injections. RESULTS: In both groups the patients showed significantly decreased WOMAC scores and Lequesne scores at 4, 12, 26 and 52 weeks after HA injections as compared with the baseline scores (all P < 0.01). No significant difference was found between the two groups in WOMAC scores and Lequesne scores at 4 or 12 weeks after the injections (both P>0.05). The WOMAC scores and Lequesne scores at 26 and 52 weeks after the injections were significantly higher in patellofemoral arthritis group than in CP group (both P < 0.05). In both groups, the 30-m walking time and the stair ascending and descending time decreased significantly at 1, 2, 3, and 4 weeks after HA injections (all P < 0.05) without significant differences between the two groups (all P>0.05). CONCLUSIONS: HA injection through the subpatellar route is effective for treatment of CP. HA injection produces better long-term efficacy for treatment of early CP than for advanced CP where patellofemoral arthritis occurs.

$H_\infty $ Control for 2-D Fuzzy Systems With Interval Time-Varying Delays and Missing Measurements.

In this paper, we consider the H∞ control problem for a class of 2-D Takagi-Sugeno fuzzy described by the second Fornasini-Machesini local state-space model with time-delays and missing measurements. The state delays are allowed to be time-varying within a known interval. The measurement output is subject to randomly intermittent packet dropouts governed by a random sequence satisfying the Bernoulli distribution. The purpose of the addressed problem is to design an output-feedback controller such that the closed-loop system is globally asymptotically stable in the mean square and the prescribed H∞ performance index is satisfied. By employing a combination of the intensive stochastic analysis and the free weighting matrix method, several delay-range-dependent sufficient conditions are presented that guarantee the existence of the desired controllers for all possible time-delays and missing measurements. The explicit expressions of such controllers are derived by means of the solution to a class of convex optimization problems that can be solved via standard software packages. Finally, a numerical simulation example is given to demonstrate the applicability of the proposed control scheme.

State estimation for a class of artificial neural networks with stochastically corrupted measurements under Round-Robin protocol.

This paper is concerned with the state estimation problem for a class of artificial neural networks (ANNs) without the assumptions of monotonicity or differentiability of the activation functions. The measured outputs are corrupted by stochastic noise signal whose intensity is quantified by a nonlinear function. In order to accommodate the bandwidth limit of the communication channel between the ANN and the state estimator, an equal allocation scheme (i.e. Round-Robin protocol) of the communication resource is employed to effectively mitigate data congestions and save energies. A set of zero-order holders (ZOHs) is utilized to store the received measurements, such that the utilization of the received measurements can be maximized. An update matrix approach is developed to handle the time-varying yet periodic time-delays resulting from the adoption of the Round-Robin protocol. The aim of the proposed problem is to design a state estimator such that the error dynamics is exponentially ultimately bounded. A combination of the Lyapunov stability theory and the stochastic analysis technique is used to derive some easy-to-test conditions for the existence of the desired state estimator. The estimator gains are characterized by the solution to a convex optimization problem that is solved via the semi-definite programme method. Simulation results are given to demonstrate the effectiveness of the proposed estimation approach.

[Iohexol tracing of hyaluronic acid distribution in the knee joint cavity of rabbits].

OBJECTIVE: To investigate the distribution of hyaluronic acid (HA) with iohexol tracing in the knee joint cavity of rabbits using CT plain scan, three-dimensional reconstruction and Χ-ray and observe how different injection sites affect HA distribution. METHODS: Mixtures of HA and iohexol (tracer) were prepared that contained final iohexol concentrations of 2.5%, 5%, 10%, 20%, or 40%. The HA-iohexol mixtures (0.5 ml) were injected into rabbit knee joints, and the optimal iohexol concentration that allowed clear differentiation of the injected agents from the surrounding tissues was determined using dual-source CT plain scan and three-dimensional reconstruction technique. The HA-iohexol mixture (0.5 ml) containing the optimal concentration of iohexol was then injected into the knees of the rabbits either through the patella medial approach or the medial joint line approach, and HA distribution in the knee joint cavity was observed using CT scan and Χ-ray. RESULTS: The CT value of HA-iohexol mixture increased progressively with the tracer concentration. After injection of the mixture containing 2.5%, 5%, 10%, 20%, and 40% iohexol, the CT value ratios of the soft tissue, HA-iohexol mixture and bone cortex were 2:7:46, 2:14:44, 2:28:44, 2:60:46, and 2:98:45, respectively, and a iohexol concentration of 5% was determined as optimal for differntiating the injected agents from the surrounding tissues. The HA-iohexol mixutre containing 5% iohexol injected through the medial-patellar approach was distributed mainly over the patello-femoral joint, and that injected through the joint line approach was found mainly over the tibio-femoral joint. CONCLUSION: HA-iohexol mixture containing 5% iohexol allows clear differentiation of bone cortex and soft tissues in rabit knee joint from the injected agents on CT scan and Χ-ray, and the injection approach can influence HA distribution in the knee joint cavity.