Ginsenoside Rh2 augmented anti-PD-L1 immunotherapy by reinvigorating CD8+ T cells via increasing intratumoral CXCL10.

Profiting from the sustained clinical improvement and prolonged patient survival, immune checkpoint blockade of programmed cell death protein 1 (PD-1)/programmed death-ligand 1 (PD-L1) axis has emerged as a revolutionary cancer therapy approach. However, the anti-PD-1/PD-L1 antibodies only achieve a clinical response rate of approximately 20%. Herein, we identified a novel combination strategy that Chinese medicine ginseng-derived ginsenoside Rh2 (Rh2) markedly improved the anti-cancer efficacy of anti-PD-L1 antibody in mice bearing MC38 tumor. Rh2 combined with anti-PD-L1 antibody (combo treatment) further triggered the infiltration, proliferation and activation of CD8+ T cells in the tumor microenvironment (TME). Depletion of CD8+ T cells by mouse CD8 blocking antibody abolished the anti-cancer effect of combo treatment totally. Mechanistically, combo treatment further increased the expression of CXCL10 through activating TBK1-IRF3 signaling pathway, explaining the increased infiltration of T cells. Employing anti- CXC chemokine receptor 3 (CXCR3) blocking antibody prevented the T cells infiltration and abolished the anti-cancer effect of combo treatment. Meanwhile, combo treatment increased the percentage of M1-like macrophages and raised the ratio of M1/M2 macrophages in TME. By comparing the anti-cancer effect of combo treatment among MC38, CT26 and 4T1 tumors, resident T cells were considered as a prerequisite for the effectiveness of combo treatment. These findings demonstrated that Rh2 potentiated the anti-cancer effect of PD-L1 blockade via promoting the T cells infiltration and activation, which shed a new light on the combination strategy to enhance anti-PD-L1 immunotherapy by using natural product Rh2.

Energy Criticality Avoidance-Based Delay Minimization Ant Colony Algorithm for Task Assignment in Mobile-Server-Assisted Mobile Edge Computing.

Mobile edge computing has been an important computing paradigm for providing delay-sensitive and computation-intensive services to mobile users. In this paper, we study the problem of the joint optimization of task assignment and energy management in a mobile-server-assisted edge computing network, where mobile servers can provide assisted task offloading services on behalf of the fixed servers at the network edge. The design objective is to minimize the system delay. As far as we know, our paper presents the first work that improves the quality of service of the whole system from a long-term aspect by prolonging the operational time of assisted mobile servers. We formulate the system delay minimization problem as a mixed-integer programming (MIP) problem. Due to the NP-hardness of this problem, we propose a dynamic energy criticality avoidance-based delay minimization ant colony algorithm (EACO), which strives for a balance between delay minimization for offloaded tasks and operational time maximization for mobile servers. We present a detailed algorithm design and deduce its computational complexity. We conduct extensive simulations, and the results demonstrate the high performance of the proposed algorithm compared to the benchmark algorithms.

A Robust Transmission Scheduling Approach for Internet of Things Sensing Service with Energy Harvesting.

Maximizing the utility under energy constraint is critical in an Internet of Things (IoT) sensing service, in which each sensor harvests energy from the ambient environment and uses it for sensing and transmitting the measurements to an application server. Such a sensor is required to maximize its utility under the harvested energy constraint, i.e., perform sensing and transmission at the highest rate allowed by the harvested energy constraint. Most existing works assumed a sophisticated model for harvested energy, but neglected the fact that the harvested energy is random in reality. Considering the randomness of the harvested energy, we focus on the transmission scheduling issue and present a robust transmission scheduling optimization approach that is able to provide robustness against randomness. We firstly formulate the transmission scheduling optimization problem subject to energy constraints with random harvested energy. We then introduce a flexible model to profile the harvested energy so that the constraints with random harvested energy are transformed into linear constraints. Finally, the transmission scheduling optimization problem can be solved traditionally. The experimental results demonstrate that the proposed approach is capable of providing a good trade-off between service flexibility and robustness.

Comparison of plasma pharmacokinetics of Tanreqing solution between intratracheal aerosolization and intravenous injection in rats.

A rapid ultra high performance liquid chromatography tandem mass spectrometry method was developed for the simultaneous analysis of baicalin, oroxylin A-7-O-ß-d-glucoronide and chlorogenic acid in rats plasma, and applied to comparison of pharmacokinetics of Tanreqing solution between intratracheal aerosolization and intravenous injection. Results of the analytical method validation assay showed high sensitivity, accuracy and suitable recovery. Results of pharmacokinetics showed similar decline phases for baicalin, oroxylin A-7-O-ß-d-glucoronide and chlorogenic acid in two different delivery routes. The half-lives of intratracheal aerosolization and intravenous injection were 0.90 and 1.22 h for baicalin, 0.47 and 0.17 h for oroxylin A-7-O-ß-d-glucoronide and 0.22 and 0.13 h for chlorogenic acid, and this implies that compounds were retained in the lung for a relatively short time. This study was the first to provide important pharmacokinetics information for traditional Chinese medicine delivery to the lung.

The Effect of Ginger Juice Processing on the Chemical Profiles of Rhizoma coptidis.

Rhizoma coptidis (RC) has been used as an herbal medicine in China for over one thousand years, and it was subjected to specific processing before use as materia medica. Processing is a pharmaceutical technique that aims to enhance the efficacy and/or reduce the toxicity of crude drugs according to traditional Chinese medicine theory. In this study, the chemical profiles of RC, ginger juice processed RC (GRC), and water processed RC (WRC) was determined to reveal the mechanism of processing of RC. UPLC-QTOF-MS analysis of methanol extract of RC, GRC, and WRC has been conducted to investigate the effect of processing on the composition of RC. HPLC-PDA was used to determine the variance of total alkaloids and seven alkaloids of RC during the processing. The volatiles of RC, GRC and ginger juice were separated by distillation, the change of volatiles content was recorded and analyzed, and the qualitative analysis of the volatiles was carried out using GC-MS. The microstructures of RC, GRC and WRC were observed using a light microscope. Results showed that ginger juice/water processing had limited influence on the composition of RC's methanol extract, but significant influence on the content of some alkaloids in RC. Ginger juice processing significantly increased (p < 0.05) the volatiles content of RC and changed the volatiles composition obviously. Processing also had an influence on the microstructure of RC. This research comprehensively revealed the mechanism of ginger juice processing of RC.

[Freeze drying process optimization of ginger juice-adjuvant for Chinese materia medica processing and stability of freeze-dried ginger juice powder].

Ginger juice， a commonly used adjuvant for Chinese materia medica， is applied in processing of multiple Chinese herbal decoction pieces. Because of the raw materials and preparation process of ginger juice， it is difficult to be preserved for a long time， and the dosage of ginger juice in the processing can not be determined base on its content of main compositions. Ginger juice from different sources is hard to achieve consistent effect during the processing of traditional Chinese herbal decoction pieces. Based on the previous studies， the freeze drying of ginger juice under different shelf temperatures and vacuum degrees were studied， and the optimized freeze drying condition of ginger juice was determined. The content determination method for 6-gingerol， 8-gingerol， 10-gingerol and 6-shagaol in ginger juice and redissolved ginger juice was established. The content changes of 6-gingerol， 8-gingerol， 10-gingerol， 6-gingerol， 6-shagaol， volatile oil and total phenol were studied through the drying process and 30 days preservation period. The results showed that the freeze drying time of ginger juice was shortened after process optimization; the compositions basically remained unchanged after freeze drying， and there was no significant changes in the total phenol content and gingerol content， but the volatile oil content was significantly decreased(P<0.05). Within 30 days， the contents of gingerol， total phenol， and volatile oil were on the decline as a whole. This study has preliminarily proved the feasibility of freeze-drying process of ginger juice as an adjuvant for Chinese medicine processing.

[Status and suggestions for adjuvant standard for Chinese materia medica processing in China].

In this paper, the status of adjuvant standard for Chinese materia medica processing in the Chinese Pharmacopoeia 2015 edition, the National Specification of Chinese Materia Medica Processing, and the 29 provincial specification of Chinese materia medica was summarized, and the the status including general requirements, specific requirements, and quality standard in the three grade official specifications was collected and analyzed according to the "medicine-adjuvant homology" and "food-adjuvant homology" features of adjuvants. This paper also introduced the research situation of adjuvant standard for Chinese materia medica processing in China; In addition, analyzed and discussed the problems existing in the standard system of adjuvant for Chinese materia medica processing, such as lack of general requirements, low level of standard, inconsistent standard references, and lack of research on the standard, and provided suggestions for the further establishment of the national standards system of adjuvant for Chinese materia medica processing.

[Preparation and evaluation of Shedan in situ forming gel based on ocular characteristics].

To develop an ophthalmic preparation of Shedan, an in situ forming gel was prepared with the formulation containing 18% of poloxamer 407 and 5% of poloxamer 188 by response surface designs plus central composite designs. The rheology results showed that LVE range gamma should limited within 0.5%, Shedan high-frequency region, and the thixotropy recovery time is less than 5 seconds. The phase transition temperature was 33.25 °C according to curve of storage modulus and loss modulus determined by temperature scanning. Surface tension and osmometer of it determined by surface tension meter and dew point osmometer were 36.43 mN · m(-1), and 320.6 mOsm · kg(-1), respectively. Fluorescein sodium was selected as the marker to monitor the corneal residence time, and the results showed that Shedan gel could prolong drug residence for 180 min. In line with zero-order kinetics, releases of muscone and salvianolic acid B in vitro depends on gels erosion. The results of rabbit ocular irritation experiments suggested that Shedan in situ forming gel was biocompatible and nonirritant. In conclusion, a novel Shedan in situ forming gel was developed and characterized for potential drug treatment of retinal vein occlusion.

[Study on preparation of Dange Mingmu in-situ forming eye gel].

OBJECTIVE: To prepare Dange Mingmu in-situ forming eye gel. METHOD: The viscosity of Dange Mingmu in-situ forming eye gel was tested by adopting poloxamer 407 and 188 as thermosensitive materials, and optimizing by uniform design. Drug release in vitro was studied using a novel membraneless model. Eye irritation experiments were performed with rabbits. The duration of residence time in rabbit eyes was observed using fluorescence tracer method. RESULT: The gelation temperature of in-situ thermosensitive gel was lowered as the P407 concentration increased. In a certain range, the gelation temperature slowly increased with the increase of P188's concentration, and the effect of P407 was greater than that of P188. The optimized concentration of P407/P188 was 19%/1%. Based the adjusted concentration, Dange Mingmu in-situ forming eye gel. was converted into freely flowing liquid below 26.9 degrees C and became gel at 34.5 degrees C after being diluted with STF. In line with zero-order kinetics, drug release in vitro depends on gel erosion. The residence time on the surface of eyes was proved to be relatively long was and nonirritant. CONCLUSION: Uniform design is available for optimizing the formulation of thermosensitive gel for eye. The gel satisfies the requirement for ophthalmic application, and is expected to be applied in clinical practice in the future.

A tRNA-derived fragment from Chinese yew suppresses ovarian cancer growth via targeting TRPA1.

Drug discovery from plants usually focuses on small molecules rather than such biological macromolecules as RNAs. Although plant transfer RNA (tRNA)-derived fragment (tRF) has been associated with the developmental and defense mechanisms in plants, its regulatory role in mammals remains unclear. By employing a novel reverse small interfering RNA (siRNA) screening strategy, we show that a tRF mimic (antisense derived from the 5' end of tRNAHis(GUG) of Chinese yew) exhibits comparable anti-cancer activity with that of taxol on ovarian cancer A2780 cells, with a 16-fold lower dosage than that of taxol. A dual-luciferase reporter assay revealed that tRF-T11 directly targets the 3' UTR of oncogene TRPA1 mRNA. Furthermore, an Argonaute-RNA immunoprecipitation (AGO-RIP) assay demonstrated that tRF-T11 can interact with AGO2 to suppress TRPA1 via an RNAi pathway. This study uncovers a new role of plant-derived tRFs in regulating endogenous genes. This holds great promise for exploiting novel RNA drugs derived from nature and sheds light on the discovery of unknown molecular targets of therapeutics.

[Preparation and characterization of niosomes of Semen Strychni alkaloids extract].

OBJECTIVE: To prepare a noisome formulation of Semen Strychni alkaloids extract with high encapsulation efficiency. METHOD: S. Strychni alkaloids were encapsulated into niosomes by pH gradient loading method. The factors influencing on the encapsulation efficiency were investigated, and formulation and preparation process of niosomes were optimised and validated. RESULT: When the drug to lipid weight ratio was under 1: 10, pH gradient in buffer solution of citric acid at 50 degrees C was more than 4.0, the niosomes (mean diameter 179.2 nm and Zeta potential -25.41 mV) were formed with encapsulation efficiency of over 86.9%. CONCLUSION: The pH gradient loading method was reliable for preparing niosomes of Semen Strychni alkaloids extract.

Targeting Strategies for Enhancing Paclitaxel Specificity in Chemotherapy.

Paclitaxel (PTX) has been used for cancer treatment for decades and has become one of the most successful chemotherapeutics in the clinic and financially. However, serious problems with its use still exist, owing to its poor solubility and non-selective toxicity. With respect to these issues, recent advances have addressed the water solubility and tumor specificity related to PTX application. Many measures have been proposed to remedy these limitations by enhancing tumor recognition via ligand-receptor-mediated targeting as well as other associated strategies. In this review, we investigated various kinds of ligands that have emerged as PTX tumor-targeting tools. In particular, this article highlights small molecule-, protein-, and aptamer-functionalized conjugates and nanoparticles (NPs), providing a promising approach for PTX-based individualized treatment prospects.

[Preparation and characterization of non-ionic surfactant vesicle of cantharidin].

OBJECTIVE: To study the preparation of cantharidin entrapped non-ionic surfactant vesicle (noisome)and evaluate its quality. METHOD: The niosome loaded with cantharidin was prepared using injection method by non-ionic surfactants as the carrier. An centrifugation separation method and HPLC analysis method of the cantharidin were established to detect the entrapment efficiency. The optimum preparation technology was established by a orthogonal experiment. The morphology, and particle size were studied to evaluate the preparation. RESULT: The average size of niosomes were (209. 8 +/- 0.5) nm. The entrapment efficiency of the CTD-NS was (27.5% +/- 2.0%) and Zeta potential was (41.5 +/- 0.65) mV. CONCLUSION: The preparation of cantharidin noisome by TweenA and SpanB is practicable and successful. These experiments can be the basement of developing targeting drug delivery system.

[Development of Silymarin nanocrystals lyophilized power applying nanosuspension technology].

OBJECTIVE: To prepare silymarin nanosuspension and lyophilized power for enhancing the dissilution of poorly soluble drugs. METHOD: The precipitation technique was adapted to produce the silymarin nanosuspensions respectivly applying Tween 80, SDS and Poloxamer188 as stabilizers. The lyophilized formula contained 5% mannitol as cryoprotectant. Particle size, Polydispersity index and Zeta potential were detected by Mastersizer nano ZS (Malvern England). Morphological character was observed with Transmission Electron Microscopy. The product's structure was performed with X-ray diffractometer. RESULT: The silymarin nanosuspension was successfully prepared, in which the drug particle size was about 100-300 nm,and the particles had ball-like shape and good dispersive properties. CONCLUSION: This study provided potential for the neotype dosage form development of the Chinese Traditional Medicine.

Simultaneous determination of seven flavonoids, two phenolic acids and two cholesterines in Tanreqing injection by UHPLC-MS/MS.

A new ultra-high performance liquid chromatography combined with triple quadrupole mass spectrometry was developed to evaluate the quality of Tanreqing injection. Seven flavonoids (Rutin, Baicalin, Scutellarin, Chrysin-7-O-Beta-d-glucoronide, Oroxylin A-7-O-ß-d-glucoronide, Wogonin, Luteolin-7-O-glucoside), two phenolic acids (Chlorogenic acid, Caffeic acid) and two cholesterines (Ursodeoxycholic acid, Chenodeoxycholic acid) in Tanreqing injection could be measured simultaneously. For the determination of the eleven compounds, the conditions were set as follows: The mobile phase was a gradient of 0.1% aqueous formic acid solution (A) and acetonitrile (B); the flow rate was 0.2 mL min-1, the column was Acquity UPLC HSS T3 column (2.1 mm × 100 mm, 1.8 µm); and the multiple-reaction monitoring (MRM) with a negative electro spray ionization interface (ESI-) was selected. Within the test ranges, all the standard regression curves showed excellent linear regression (r > 0.99). In terms of (relative standard deviation) RSDs, the precision, repeatability and stability of the eleven compounds were all lower than 3%. The recovery rates of Tanreqing injection and the RSD were 97.8-103.7% and 0.4%-2.0%, respectively. The RSD value was in accordance with the requirements of less than 3.0%. This method has been successfully used in the analysis of Tanreqing injection. In summary, a fast, accurate and reliable UPLC-ESI--MS/MS method was successfully developed for the simultaneous detection of the eleven major active ingredients with different chemical structures in Tanreqing injection, and can be used for the quality control of Tanreqing injection as well.

[Optimal extraction of nuciferine and flavone from lotus leaf based on central composite design and response surface methodology].

OBJECTIVE: To optimize the process of extracting effective constituents from lotus leaf. METHOD: Independent variables were ethanol concentration reflux time and solvent fold, dependent variables were extraction rates of nuciferine and flavone in lotus leaf, central composite design and response surface methodology were used for optimization of extraction of lotus leaf. RESULT: The optimum conditions of extraction process were 75% -80% ethanol, 2-3 hours for reflux, 20-25 fold solvent and 2 times for extraction. Bias between observed and predicted of rates of nuciferine and flavone values were 5.53%, -6.02%, respectively. CONCLUSION: The values observed and predicted were close to each other, which proved that the optimization of of extraction of lotus leaf by central composite design and response surface methodology was reasonable and successful.

[Comparative study on in vitro drug-release between Tuizhang ophthalmic gel and Tuizhang oculentum].

OBJECTIVE: To prepare tuizhang gel to cure cataract, the characteristics of Tuizhang gel on the drug-releasing in vitro were evaluated by compared with Tuizhang oculentum. METHOD: The emodin and chrysophanol in the releasing mediator were determined by HPLC, and the drug releasing characteristics of Tuizhang gel and Tuizhang oculentum were studied by bag filter method. RESULT: The emodin and chrysophanol in Tuizhang gel released (98.3, 1.1)%, (95.8, 1.8)% within 24 hours, respectively, while those in tuizhang oculentum released (10.62, 0.7)%, (10.46, 0.4)%, respectively. The emodin and chrysophanol in Tuizhang gel released more quickly and completely than in Tuizhang oculentum, so Tuizhang gel has enhanced the bioavailability. CONCLUSION: The Tuizhang gel is characterized by slow-release to some degree, and it shows a promising future on ophthalmic drug delivery.

Self-Assembled Thermoresponsive Nanogels Prepared by Reverse Micelle → Positive Micelle Method for Ophthalmic Delivery of Muscone, a Poorly Water-Soluble Drug.

This study aimed to design a nanocarrier ophthalmic delivery system of muscone, a poorly water-soluble drug. The muscone thermoresponsive nanogels were self-assembled by reverse micelle → positive micelle method. Muscone was demonstrated to have uniform narrow particle size distribution in nanogel by the dynamic light scattering test. The developed nanocomposite hydrogel had a high muscone loading, and the rheology results showed that the phase transition temperature was 34.05°C. Thixotropy test indicated that the nanogel was able to resist the blinking of eyes because of the thixotropy recovery time, which is <5 s. Compared with muscone eye drops, muscone nanogels showed longer retention time on the corneal surface using fluorescent labeling technology and produced a 3.4-fold increase in apparent permeability coefficients (Papp). Draize testing showed that the developed nanogel caused no eye irritation. In vivo pharmacokinetic study indicated that the nanogel could significantly increase the bioavailability of muscone after administration compared with eye drops. These results indicate that self-assembled thermoresponsive nanogel prepared by reverse micelle → positive micelle method has potential for the ophthalmic delivery of poorly water-soluble drugs.

Chemical profiling and cytotoxicity assay of bufadienolides in toad venom and toad skin.

ETHNOPHARMACOLOGICAL RELEVANCE: Toad venom and toad skin have been widely used for treating various cancers in China. Bufadienolides are regarded as the main anticancer components of toad venom, but the difference on composition and anticancer activities of bufadienolides between toad venom and toad skin remains unclear. METHODS: Fractions enriched with free and conjugated bufadienolides were prepared from toad venom and toad skin. Bufadienolides in each fraction were comprehensively profiled by using a versatile UHPLC-TOF-MS method. Relative contents of major bufadienolides were determined by using three bufogenins and one bufotoxin as marker compounds with validated UHPLC-TOF-MS method. Furthermore, cytotoxicity of the fractions was examined by MTT assay. RESULTS: Two fractions, i.e., bufogenin and bufotoxin fractions (TV-F and TV-C) were isolated from toad venom, and one bufotoxin fraction (TS-C) was isolated from toad skin. Totally 56 bufadienolides in these three fractions were identified, and 29 were quantified or semi-quantified. Bufotoxins were identified in both toad venom and toad skin, whereas bufogenins exist only in toad venom. Bufalin-3-conjugated bufotoxins are major components in toad venom, whereas cinobufotalin and cinobufagin-3-conjugated bufotoxins are main bufotoxins in toad skin. MTT assay revealed potent cytotoxicity of all the fractions in an order of TV-F>TV-C>TS-C. CONCLUSIONS: Our study represents the most comprehensive investigation on the chemical profiles of toad venom and toad skin from both qualitative and quantitative aspects. Eight bufotoxins were identified in toad skin responsible for the cytotoxicity for the first time. Our research provides valuable chemical evidence for the appropriate processing method, quality control and rational exploration of toad skin and toad venom for the development of anticancer medicines.

[Studies on preparation technology of Flos Magnoliae essential oil-beta-cyclodextrin inclusion complex].

OBJECTIVE: To find the best condition of the preparation technology of Flos Magnoliae essential oil-beta-cyclodextrin inclusion complex. METHOD: L9(3(4)) table was used to examine the effects of 4 factors, and the inclusion rate of each test was determined of orthogonal test. RESULT: The best condition was:oil:beta-cyclodextrin:water = 1:8:60 (mL:g:mL), stirring for 1 hour at 80 degrees C. CONCLUSION: The complex prepared on the condition aforementioned is stable and stirring has a highest inclusion rate.