Novel indocyanine green-loaded photothermal nanoparticles targeting TRPV1 for thermal ablation treatment of severe murine asthma induced by ovalbumin and lipopolysaccharide.

To identify a replacement strategy for bronchial thermoplasty (BT) with non-invasive and free-of-severe side effect is urgently needed in the clinic for severe asthma treatment. In this study, PLGA-PEG@ICG@TRPV1 pAb (PIT) photothermal nanoparticles targeting bronchial TRPV1 were designed for photothermal therapy (PTT) against severe murine asthma induced by ovalbumin and lipopolysaccharide. PIT was formulated with a polyethylene glycol (PEG)-grafted poly (lactic-co-glycolic) acid (PLGA) coating as a skeleton structure to encapsulate indocyanine green (ICG) and was conjugated to the polyclonal antibody against transient receptor potential vanilloid 1 (TRPV1 pAb). The results revealed that PIT held good druggability due to its electronegativity and small diameter. PIT demonstrated great photothermal effects both in vivo and in vitro and exhibited good ability to target TRPV1 in vitro because of its selective cell uptake and specific cell toxicity toward TRPV1-overexpressing cells. The PIT treatment effectively reduced asthma symptoms in mice. This is evident from improvements in expiratory airflow limitation, significant decreases in inflammatory cell infiltration in the airways, and increases in goblet cell and columnar epithelial cell proliferation. In conclusion, PIT alleviates severe murine asthma symptoms through a combination of TRPV1 targeting and photothermal effects.

Transcriptome and metabolome profiling of the medicinal plant Veratrum mengtzeanum reveal key components of the alkaloid biosynthesis.

Veratrum mengtzeanum is the main ingredient for Chinese folk medicine known as "Pimacao" due to its unique alkaloids. A diverse class of plant-specific metabolites having key pharmacological activities. There are limited studies on alkaloid synthesis and its metabolic pathways in plants. To elucidate the alkaloid pathway and identify novel biosynthetic enzymes and compounds in V. mengtzeanum, transcriptome and metabolome profiling has been conducted in leaves and roots. The transcriptome of V. mengtzeanum leaves and roots yielded 190,161 unigenes, of which 33,942 genes expressed differentially (DEGs) in both tissues. Three enriched regulatory pathways (isoquinoline alkaloid biosynthesis, indole alkaloid biosynthesis and tropane, piperidine and pyridine alkaloid biosynthesis) and a considerable number of genes such as AED3-like, A4U43, 21 kDa protein-like, 3-O-glycotransferase 2-like, AtDIR19, MST4, CASP-like protein 1D1 were discovered in association with the biosynthesis of alkaloids in leaves and roots. Some transcription factor families, i.e., AP2/ERF, GRAS, NAC, bHLH, MYB-related, C3H, FARI, WRKY, HB-HD-ZIP, C2H2, and bZIP were also found to have a prominent role in regulating the synthesis of alkaloids and steroidal alkaloids in the leaves and roots of V. mengtzeanum. The metabolome analysis revealed 74 significantly accumulated metabolites, with 55 differentially accumulated in leaves compared to root tissues. Out of 74 metabolites, 18 alkaloids were highly accumulated in the roots. A novel alkaloid compound viz; 3-Vanilloylygadenine was discovered in root samples. Conjoint analysis of transcriptome and metabolome studies has also highlighted potential genes involved in regulation and transport of alkaloid compounds. Here, we have presented a comprehensive metabolic and transcriptome profiling of V. mengtzeanum tissues. In earlier reports, only the roots were reported as a rich source of alkaloid biosynthesis, but the current findings revealed both leaves and roots as significant manufacturing factories for alkaloid biosynthesis.

De Novo Biosynthesis of the Oleanane-Type Triterpenoids of Tunicosaponins in Yeast.

Tunicosaponins are natural products extracted from Psammosilene tunicoides, which is an important ingredient of Yunnan Baiyao Powder, an ancient and famous Asian herbal medicine. The representative aglycones of tunicosaponins are the oleanane-type triterpenoids of gypsogenin and quillaic acid, which were found to manipulate a broad range of virus-host fusion via wrapping the heptad repeat-2 (HR2) domain prevalent in viral envelopes. However, the unknown biosynthetic pathway and difficulty in chemical synthesis hinder the therapeutic use of tunicosaponins. Here, two novel cytochrome P450-dependent monooxygenases that take part in the biosynthesis of tunicosaponins, CYP716A262 (CYP091) and CYP72A567 (CYP099), were identified from P. tunicoides. In addition, the whole biosynthesis pathway of the tunicosaponin aglycones was reconstituted in yeast by transforming the platform strain BY-bAS with the CYP716A262 and CYP716A567 genes, the resulting strain could produce 146.84 and 314.01 mg/L of gypsogenin and quillaic acid, respectively. This synthetic biology platform for complicated metabolic pathways elucidation and microbial cell factories construction can provide alternative sources of important natural products, helping conserve natural plant resources.

The complete plastid genome sequence of Neopicrorhiza scrophulariiflora (Plantaginaceae): an endangered species endemic to The Himalayas regions.

Neopicrorhiza scrophulariiflora (Pennell) Hong, an endangered perennial species, is endemic to the Eastern Himalayas and Hengduan Mountains. In this study, we have sequenced the complete chloroplast genome of N. scrophulariiflora, which is 152,643 bp in length, including two inverted repeat (IR, 25,829 bp) regions, one large single copy region (LSC) and one small single copy region (SSC) of 83,191 bp and 17,794 bp, respectively. The cp genome has 131 annotated genes, including 86 protein-coding genes, 37 tRNA genes, and eight rRNA genes. The overall GC content of it is 38.1%. Phylogenetic analysis using total chloroplast genome DNA sequence of 14 species revealed that N. scrophulariiflora was closely relates to two species of Veronica with 100% bootstrap value.

[Frequency-Selectable RF Ablation Treatment System].

In order to support study of new radiofrequency ablation treatments and finally realize of precise conformal ablation of targeted tissue, this paper proposed a system which is able to generate radio frequency currents at different frequencies. The designed system bases on the basic principle of Class E power amplifier and uses MSP430 microcontroller as a controller and uses a touch screen as human-machine interface to design a frequency-selectable radiofrequency ablation treatment system,. The RF signals at frequencies of 230 kHz, 460 kHz, 920 kHz, and 2 000 kHz were tested to verify the feasibility of the system, and by using a tissue phantom, tested the heating ability of the RF signals at frequencies of 460 kHz, 920 kHz, and 2 000 kHz. The results show the device could well heat the tissue at the three frequencies. The study has also found that both the highest temperature and the ablation area decreases with RF frequency, when using the constant power control mode.

Cryo-thermal therapy elicits potent anti-tumor immunity by inducing extracellular Hsp70-dependent MDSC differentiation.

Achieving control of metastatic disease is a long-sought goal in cancer therapy. Treatments that encourage a patient's own immune system are bringing new hopes in reaching such a goal. In clinic, local hyperthermia and cryoablation have been explored to induce anti-tumor immune responses against tumors. We have also developed a novel therapeutic modality of cryo-thermal treatment by alternating liquid nitrogen (LN2) cooling and radio frequency (RF) heating, and better therapeutic effect was achieved in treating metastatic cancer in animal model. In this study, we investigated the mechanism of systemic immune response elicited by cryo-thermal therapy. In the 4T1 murine mammary carcinoma model, we found that local cryo-thermal therapy resulted in a considerable reduction of distant lung metastases, and improved long-term survival. Moreover, results of tumor re-challenge experiments indicated generation of a strong tumor-specific immune memory after the local treatment of primary tumors. Our further study indicated that cryo-thermal therapy caused an elevated extracellular release of Hsp70. Subsequently, Hsp70 induced differentiation of MDSCs into mature DCs, contributing to the relief of MDSCs-mediated immunosuppression and ultimately the activation of strong anti-tumor immune response. Our findings reveal new insight into the mechanism of robust therapeutic effects of cryo-thermal therapy against metastatic cancers.

[Isolation and diversity analyses of endophytic fungi from Paris polyphylla var. yunnanensis].

The paper is aimed at studying the diversity of endophytic fungi community from Paris polyphylla var. yunnanensis, and to provide a scientific basis for the utilization value of the endophytic fungi as bioactive material resources. In the present study, endophytic fungi were isolated from roots, rhizomes and leaves of wild P. polyphylla var. yunnanensis collected from Baoshan, Heqing county and Songming city of Yunnan province, and identified and classified by morphological methods together with its ITS sequence analysis. Seven and forty-nine strains of endophytic fungi were isolated from P. polyphylla var. yunnanensis. They were identified belonging to 41 genus. In these 41 genus, 3 genus exist in root only, 12 genus only exist in rhizome and 8 genus only exist in leaf. There was difference in endophytic fungi isolated from different sample sites. Endophytic fungi diversity from rhizomes of Heqing site was the highest. Endophytic fungi similarity coefficient was low among different sites and tissues. Based on these results, it is reasonable to propose that endophytic fungi of P. polyphylla var. yannanensis from different tissue and different sample sites has a certain difference which is possibly relate to their different habitats, different structure and composition of each tissue.

Study of tumor growth under hyperthermia condition.

The new concept of keeping primary tumor under control in situ to suppress distant foci sheds light on the treatment of metastatic tumor. Hyperthermia is considered as one of the means for controlling tumor growth. To simulate the tumor growth, a continuum mathematical model has been introduced. The newest understanding of the Warburg effect on the cellular metabolism and diffusion of the nutrients in the tissue has been taken into consideration. The numerical results are compared with the in vivo experimental data by fitting the tumor cell doubling time/tumor cell growth rate under different thermal conditions. Both the tumor growth curve and corresponding average glucose concentration have been predicted. The numerical results have quantitatively illustrated the controlling effect on tumor growth under hyperthermia condition in the initial stage.

Design of microprobe for accurate thermal treatment of tumor.

Thermal treatment has become an alternative modality for cancer treatment. Low temperature freezing and high temperature heating kill tumor cells effectively through direct and indirect injuries by biochemical and physical stresses. Hyperthermia at a mildly elevated temperature has also been reported to induce biochemical alternations to kill tumor cells and to stimulate immunological response to prevent metastasis. The comprehensive multi-scale biological responses to different thermal history experienced demand an accurate temperature control of the thermal system used for such a treatment. A thermal system was built in our lab utilizing RF heating and liquid nitrogen cooling through a needle probe. In practice, difficulties involved in temperature measurement for in vivo monitoring and control of thermal input through two-phase LN2 flow inside the probe compromise the treatment outcome. To ensure an accurate temperature control, a new model was developed to study the dynamic freezing capacity of the cryo-probe by accounting for the probe shape and dimensions. The model was validated by experiments and used to predict the freezing processes under different conditions. Numerical simulation results showed that combined with RF heating, the system could be used to perform different treatment protocols with an accurate temperature control.

Design of a new probe for tumor treatment in the alternate thermal system based on numerical simulation.

A new probe for tumor treatment is designed and simulated in this study. This probe combines the cryosurgery and hyperthermia which is suitable for the treatment of subcutaneous tumors. Simulations of the cooling and heating processes demonstrate that the probes are capable of treating the tumor effectively. And the numerical results indicate that the lengths of the probe, the diameters of the inner tube and the pressures of liquid nitrogen influence the probes' cooling ability. The temperature responses at the tumor base induced by different probes are similar, though the great differences appear on the treatment interface of the probes, thus the temperature gradient within the tumor. Based on the simulation results, the heating effect of the probe is shown to be effective in damaging the tumor while protecting normal tissue in the surrounding. Animal experiments will be carried out using this type of probe to treat tumor in the near future.

Women's cancers in developing countries: from research to an integrated health systems approach.

The article focuses on two women's cancers, breast and cervical cancer, that are much more deadly in developing countries than in developed countries. Early detection can make a significant difference for the treatment outcome of these two cancers and there are now cost-effective tools for prevention and screening. The authors propose a new public health approach to these two cancers in developing countries where resources for effective cancer control are very limited and offer a framework for putting women's cancers in developing countries on the global public health agenda. The key areas are: 1. Proposals for a new, integrated public health approach to women's cancers (breast and cervical) in resource poor settings; 2. Reviews of the evidence for cost-effective screening and early detection of breast and cervical cancer, and discussion of some of the lessons learned from HIV/AIDS on an integrated health systems approach; 3. Outlines of ways to make a priority of women?s cancers in developing countries on the political agenda of international agencies.

Study on tumor microvasculature damage induced by alternate cooling and heating.

Tumor vasculature damage induced by various thermal treatments has been studied in vivo via laser confocal microscopy. Murine mammary carcinoma 4T1 was implanted in the nude mice dorsal skin fold window chamber. The implanted tumor was treated by alternate cooling and heating. Results showed that the treatment was much more effective as compared with that of cooling or heating alone, especially in damaging the tumor vasculature. In general, tumor vascular response to thermal stimuli was heterogeneous. All the treatments of hyperthermia at 42 degrees C (for 1 h), alternate cooling at 1 degrees C and heating at 42 degrees C (for 1/2 h each) and that of -10 degrees C/42 degrees C (for 1/2 h each) enhanced liposome extravasation. Pre-cooling tumor at 1 degrees C preserved most of the vascular integrity but partially inhibited the effect of post-hyperthermia at 42 degrees C. On the other hand, cooling at -10 degrees C for 1/2 h before heating at 42 degrees C caused severe vessel damage. Histo-pathological analyses further confirmed the effect as rare tumor vessel recurrence and large necrotic tumor tissue areas shown on the 7th day after the treatment.

Immunological response induced by alternated cooling and heating of breast tumor.

A new in-situ thermal physical method combining both cryosurgery and local hyperthermia was used to treat mice bearing 4T1 murine mammary carcinoma. The induced anti-tumor immune response was investigated. The cryo/heat treatment resulted in stimulation of CTL response and attraction of immunocytes into the tumor debris, which correlated well to the tumor rejection in re-implantation. The results suggested that alternated cooling and heating had synergistic effect and might be developed into an alternative modality for tumor therapy.

Enhanced efficacy of anti-tumor liposomal doxorubicin by hyperthermia.

The efficiency of novel tumor chemotherapeutics could be increased using targeted drug delivery by hyperthermia. In this paper, the 3D liposomal doxorubicin distribution in the tumor tissue enhanced by local hyperthermia was quantitatively studied in real time using laser confocal microscopy. Results showed that the thermally induced liposomal doxorubicin extravasation was non-uniform and more excessive in the peripheral region than that in the tumor center. The effect of the thermally targeted drug delivery was also investigated. On the 1st, 3rd day after the thermally targeted drug treatment, histological examination showed that many nucleolus were condensed and collapsed in the peripheral region. But, in the tumor center, there were no such changes found until the 3rd day. While on the 6th day, tumor cells in both the peripheral and center region were found necrotic. The enhancement of the nanoparticle anti-tumor drug effect was significant. A theoretical analysis of liposomal doxorubicin diffusion to the tumor cells in vivo was performed. Results showed that it took more than 40 hrs for the doxorubicin to get into the tumor cells in the center region from the periphery region. The theoretical results well explained the experimental observations.