Evaluation of thermotherapy against Huanglongbing (citrus greening) in the greenhouse

Huanglongbing （HLB, or citrus greening） is the most destructive disease of citrus, which is associated with Candidatus Liberibacter asiaticus （Las）. Few management options are available, aside from preventive measures such as removing infected plants, planting disease-free seedlings, and managing the insect vector. In this study, we assessed the efficacy of thermotherapy against HLB under controlled greenhouse conditions. A total of 60 two-year-old, graft-infected Citrus reticulata Blanco plants were used. The plants were randomly divided into three groups （45℃, 48℃, and untreated control）, with five plants/replicate （rep） and four reps/treatment. The treated plants were placed in phytotrons for a 4-h treatment session, repeated once per week for three consecutive weeks. Disease remission was observed eight weeks post-treatment. Real-time PCR assays revealed that Las titers in HLB-affected seedlings were significantly reduced in both 45 and 48℃ treatments four weeks after treatment, with the exception of eight plants. In contrast, Las titers in the untreated control plants increased significantly during the same period, with a maximum increase of 28-fold. Except for seven plants, Las titers in the new flushes of treated plants decreased more than 90% eight weeks after treatment. Las titers in mature leaves of treated plants decreased 56 and 60% in average at 45 and 48℃, respectively, eight weeks after treatment. The HLB symptoms and Las titer of seedings were markedly alleviated eight weeks after treatment in both 45 and 48℃ treatments. Our results laid a good foundation for the further development of citrus free-disease seedling cultivation and Huanglongbing control in the field. The whole plants were replaced for scion or branch in previous as the research object in this study, and the expression of Huanglongbing symptoms combined with real-time polymerase chain reaction （PCR） were used to evaluate the effect of heat treatment in the greenhouse.

Effect of pre-culture on virus elimination from in vitro apple by thermotherapy coupled with shoot tip culture

We evaluated the role of pre-culture on survival rate of in vitro apple plants treated by thermotherapy. Two apple cultivars, Malusxdomestica cv. Pink Lady and Huafu, were used in the experiment and both have widely grown in China and infected with Apple chlorotic leafspot virus （ACLSV） and Apple stem grooving virus （ASGV）. Results in growth and virus titer of apple plants did not exhibit clear trends during five different periods of pre-culture. Whilst, pre-culture increased the survival rate of the two cultivars during thermotherapy. The survival rate of plants pre-cultured for 13 d （P-13d） was 14 and 51% higher than that of P-ld plants for Pink Lady and Huafu, respectively. Moreover, pre-culture positively influenced regeneration of Huafu plants. The average survival rate of plants regenerated from P-ld and P-4d was 20% lower than that regenerated from P-7d, P-10d, and P-13d. The efficiency of virus eradication was determined by reverse-transcription PCR with two primer pairs for each virus, and the detection results showed that pre-culture scarcely affected apple virus elimination. Despite the fact that the two viruses were hardly detected at 5 d of thermotherapy, no virus-free plants were found in the two cultivars of regenerated apple plantlets after 30-d treatment.

Laser-induced Interstitial Thermotherapy via a Single-needle Delivery System: Optimal Conditions of Ablation, Pathological and Ultrasonic Changes

Summary： This study aimed to examine the optimal conditions of laser-induced interstitial ther- motherapy （LITT） via a single-needle delivery system, and the ablation-related pathological and ultra- sonic changes. Ultrasound （US）-guided LITT （EchoLaser system） was performed at the output power of 2--4 Wattage （W） for 1-10 min in ex vivo bovine liver. Based on the results of the ex vivo study, the output power of 3 and 4 W with different durations was applied to in vivo rabbit livers （n=24）, and VX2 tumors implanted in the hind limbs of rabbits （n=24）. The ablation area was histologically determined by hematoxylin-eosin （HE） staining. Traditional US and contrast enhanced ultrasound （CEUS） were used to evaluate the treatment outcomes. The results showed： （1） In the bovine liver, ablation disruption was grossly seen, including a strip-like ablation crater, a carbonization zone anteriorly along the fiber tip, and a surrounding gray-white coagulation zone. The coagulation area, 1.2 cm in length and 1.0 cm in width, was formed in the bovine liver subjected to the ablation at 3 W for 5 min and 4 W for 4 rain, and it extended slightly with the ablation time. （2） In the rabbit liver, after LITT at 3 W for 3 min and more, the coagulation area with length greater than or equal to 1.2 cm, and width greater than or equal to 1.0 cm, was found. Similar coagulation area was seen in the implanted VX2 carcinoma at 3 W for 5 min. （3） Gross examination of the liver and carcinoma showed three distinct regions： ablation cra- ter/carbonization, coagulation and congestion distributed from the center outwards. （4） Microscopy re- vealed four zones after LITT, including ablation crater/carbonization, coagulation, edema and conges- tion from the center outwards. A large area with coagulative necrosis was observed around a vessel in the peripheral area with edema and hyperemia. （5） The size of coagulation was consistent well to the CEUS findings. It was concluded that EchoLaser system at low power can produce a coagulation area larger than 1.0 cm×1.0 cm during a short time period. The real-time US imaging can be used to effec- tively guide and assess the treatment.

Beneficial horticultural responses from the application of solar thermotherapy to mature Huanglongbing-affected citrus trees

The detrimental effects of Huanglongbing(HLB)on citrus are well known and so is the need for effective methods to combat this disease.Solar thermotherapy(ST),one of the management methods to help alleviate some of the negative effects of HLB,is the process of heating trees to therapeutic temperatures by encompassing the tree within a plastic structure(Visqueen)to harness the sun’s natural energy.ST was applied to mature‘Valencia’citrus trees in three locations.Tree vigor,yield,and Candidatus Liberibacter asiaticus(Las)titer in leaves were monitored for two consecutive seasons post ST treatment,while fruit and juice quality were evaluated at the end of the second season.ST promoted an increase in canopy density in most groves tested and did not have a significant effect on fruit quantity,despite the prolonged exposure of the trees to increased temperatures.Moreover,Las titer was reduced in both leaves and juice processed from affected trees post ST.With respect to fruit quality,the ratio of total soluble solids to titratable acidity was higher post ST compared to controls as was the sucrose content of the juice in most treated groves,whereas limonin was higher in juice from control trees.Principal components analysis of aroma volatiles showed significant differences between juice from treated versus control trees with volatiles imparting top-note and freshness to orange juice,such as acetaldehyde,hexanal,Z-3-hexenol and linalool,being higher in juice fromtreated trees.Taste panels confirmed that flavor differences existed,ultimately showing that ST resulted in improved juice flavor in well-managed groves.Although the effects of ST on the trees appear temporary,such outcomes emphasize its utility when used as part of an integrated management strategy for HLB-affected citrus,imparting beneficial horticultural responses with minimal to positive effects on subsequent juice flavor.

Elimination of grapevine fleck virus and grapevine rupestris stem pitting-associated virus from Vitis vinifera 87-1 by ribavirin combined with thermotherapy

Vitis vinifera 87-1 plants infected by grapevine fleck virus(GFkV) and grapevine rupestris stem pitting-associated virus(GRSPaV) were used as the plant materials for virus elimination treatment. This study evaluated the effects of ribavirin at different concentrations(15 and 25 μg mL^(–1);R15 and R25, respectively), thermotherapy(37°C;T), and the combination of ribavirin and thermotherapy(R15+T and R25+T) on eliminating viruses from grapevine plants in vitro. Both R15 and R25 had phytotoxic effects and weakened plant growth. Thermotherapy positively affected the growth of grapevine plants. Plant height was significantly greater in T, R15+T, and R25+T than in CK, R15 and R25. The proportion of dead plants after T, R15+T, and R25+T was 51.4, 11.4, and 8.6%, respectively. The survival rates of regenerated plants after all treatments were >68.0%. Ribavirin concentration and treatment time were related to the regeneration of shoot tips and elimination efficiencies of the two viruses. The survival rates of plants after R15+T for 30, 40, and 50 days were 97.3, 90.7, and 74.4%, respectively. The elimination rates of GRSPaV from plants in the three time quantum were 55.6, 84.6, and 93.8%, respectively. The elimination rate of GFkV was 23.9% higher in R25(35/44) than in R15(25/45), and that of GRSPaV was 7.0% higher in R25 than in R15. The combination of thermotherapy and chemotherapy was found to have a positive effect on the eradication of GFkV and GRSPaV, and R25+T for 50 days was able to completely eliminate the two viruses from in vitro grapevines.

Application of Ansys Program in Laser-induced Interstitial Thermotherapy (LITT)

The influence of heat source by laser irradiating on tissue temperature was simulated with the Ansys program, and the distribution of tissue temperature under the heat source was shown. Compared with others, the simulation method with aasys program is more intuitionistic and can offer very good reference for Laser-induced iaterstitial thermotherapy.

Transrectal microwave thermotherapy causing a short-time influence on sperm quality in Chinese chronic nonbacterial prostatitis patients

Chronic prostatitis can affect the sperm＇s quality. Previous studies have shown that transrectal microwave thermotherapy （TRMT） results in symptomatic relief in patients with chronic prostatitis, but the effects on sperm have not been carefully investigated. This study evaluates the impact of TRMT on the relief or decrease of symptoms and quality of sperm when used to treat patients with chronic nonbacterial prostatitis. Sixty patients were enrolled in the study. TRMT treatment was administered over 5 days, 1 h per day. Semen examination was carried out pretreatment and immediately at the conclusion of the 5-day treatment. Also, it was repeated 1 month, 3 months, and 6 months later. The treatment＇s symptom relief efficacy was evaluated using the National Institutes of Health Chronic Prostatitis Symptom Index （NIH-CPSI）. After the treatment, the overall NIH-CPSI scores were lower compared to those of pretreatment. In addition, the white blood cells and lecithin in expressed prostatic secretion were normal after the treatment. The sperm count was decreased by 23.8% 3 months after the treatment, sperm motility was reduced by 10.3% immediately after treatment, and sperm deformity was increased by 17.2%. The sperm volume and PH were not affected. However, the sperm quality recovered after treatment and the malformation rate was also lower at 6 months after treatment. TRMT is a favorable and safe treatment option for patients with nonbacterial chronic prostatitis. It could relieve the patient＇s symptoms and impact on sperm quality in the short-term.

Treatment of Advanced Non-small Cell Lung Cancer with Extracorporeal High Frequency Thermotherapy Combined with Chinese Medicine

Objective： To observe the clinical efficacy and benefit response of extracorporeal high frequency thermotherapy （EHFT） combined with Chinese medicine （CM） in the treatment of patients with advanced non-small cell lung cancer. Methods： The study adopted a prospective, small sample and randomized controlled method, and the advanced non-small cell lung cancer patients were assigned to two groups according to the table of random digits, one having the treatment of EHFT combined with CM （the treatment group）, the other only with CM （the control group）. The patients in the treatment group were treated with EHFT one hour once per day, together with CM differentiation decoction, 250 mL orally taken, twice daily for 14 days as one cycle, and 3-4 cycles was performed. The patients in the control group were treated only with CM differentiation decoction using the same dose as the treatment group. The efficacies were evaluated after three to four cycles of treatment. Primary endpoints were disease control rate （DCR） and time to progression （TTP）. Secondary endpoints were overall survival time and 1-year survival rate. Results： Sixty-six patients accomplished the study. After the patients underwent different treatments, none of the patients got a complete response or partial response in both groups. In the treatment group, DCR was 72.2%, and 10 had progression of disease （28.8%）, while the DCR of the control group was 63.3%, and 11 had progression of disease （36.7%）; there was a significant statistical difference （P0.05）, suggesting that the combined regimen had superiority on the DCR. As for long-term efficacy, the median survival time （MST） of the treatment group was 7.5 months, TTP was 5.5 months, and 1-year survival rate was 21.4 %; in the control group, the results were 6.8 months, 4.5 months and 16.6% respectively. There was significant statistical difference on TTP （P0.05）, but no difference on MST or 1-year survival rate. Conclusion： EHFT combined with CM differentiation has better tolerance and short-term efficacy in the treatment of patients with advanced NSCLC.

Clinical Study on Treatment of Large Metastatic Cervical Lymphnodes of Nasopharyngeal Carcinoma by Shenlong Liquid in Combination with Thermotherapy and Radiotherapy

Objective:To observe the clinical treatment of 105 patients of nasopharyngeal carcinoma withlarge metastatic cervical lymphnodes. Methods' The patients were divided randomly into three groups, 35 foreach group. Group A was treated by combined therapy of Shenlong Liquid (SLL) with thermo- and radio-therapy, group B with combined therapy of thermo- and radio-therapy and group C with radiotherapy alone. Theclinical conditions of the three groups were similar before treatment. Results: At the end of the treatment, theimmediate regression rates of cervical lymphnodes of A, B and C group were 68. 57 %, 60. 00 % and 42. 86 %respectively, and the effect of group A was the best. In comparison of group A and C, the difference was significant (P＜0. 05), but the comparison of group B with A, or B with C was insignificant (P＞0. 05). Conclusions: The combination therapy of SLL with thermo- and radio-therapy was preferable in treating largemetastatic lymphnodes, as SLL was beneficial in increasing the sensitivity of tumor to thermotherapy, reducingtheir side-effect and preventing the hematogenous metastasis.

Breast Cancer Hyperthermia Treatment Based on Slotted Patch Antenna at 2.45 GHz

The present work proposed a simple model for breast cancer hyperthermia treatment at 2.45 GHz. The proposed model involves nine-element antennas alongside a numerical breast comprising multiple tumors. Using a coupled EM-Thermal simulation in the CST suite, the simulated results for a single antenna showed a reflection coefficient (S11) better than -47 dB and demonstrated a bandwidth of 78 MHz. The specific absorption rate (SAR) as a function of input powers was examined inside the breast tissues, where it exhibited a promising performance higher than 3 W/kg at the tumor volume when the applied power was at a reasonable level of 1.5 W whereas it was well attained under the recommend IEEE level of 1.6 W/kg through the surrounded health tissues. Taking into consideration nine-element antennas covering the breast containing two different located tumors, the maximum temperature as a function of treatment time was presented at which a resulting temperature of 43°C was obtainable within 10 minutes, favored for hyperthermia purposes. Considering the maximum power level of 1.5 W, the potential use of applying three-element antennas, simultaneously with 0.5 W, could be achieved.

Apple stem grooving virus is associated with leaf yellow mottle mosaic disease on Citrus grandis cv. Huangjinmiyou in China

Although it is usually latent on citrus, apple, and pear, apple stem grooving virus(ASGV) poses a great risk to many sensitive cultivars. Since severe leaf yellow mottle mosaic(LYMM) symptoms have been observed on Huangjinmiyou(HJY) pummelos(Citrus grandis cv. Huangjinmiyou), a commercial variety that is widely cultivated in South China, high throughput sequencing(HTS) was used to find potential pathogens and only three divergent ASGV variants were identified. The three ASGV variants shared 81.03–82.34% genome-wide pairwise identities with each other, and were separately closest to other ASGV variants from different hosts and/or geographical regions, as indicated by viral phylogenies. However, these new variants may have developed from viral interstrain interactions, based on the results of recombination analysis. A large-scale survey using reverse transcription-PCR(RT-PCR) protocols designed for the three ASGV variants revealed a high incidence(92.7–100%) of ASGV in symptomatic HJY trees from 11 major citrusproducing regions in China. None of ASGV were detected in asymptomatic trees. Temperature treatments applied to the symptomatic HJY plants showed that ASGV is sensitive to high temperatures(30–35°C), at which not only the plants recovered, but also the viruses were not detected by RT-PCR, while at low temperatures(20–24°C), both the symptoms and viruses remained detectable. These data show that ASGV is associated with the LYMM disease prevalent on HJY in China, and this is the significant basis especially of taking appropriate measures timely to manage the disease.

Treatment review of sight threatening circumscribed choroidal haemangioma

AIMTo describe our clinical experience in treating circumscribed choroidal haemangioma (CCH) in a tertiary referral centre over a fifteen year period prior to photodynamic therapy.

Photothermal Therapy Using TiO2 Nanotubes in Combination with Near-Infrared Laser

We report the in vitro cell test and in vivo animal test results of titanium oxide nanotubes (TiO2 NTs) as a potential therapeutic agent used for cancer thermotherapy in combination with near-infrared (NIR) laser. The in vitro cell test results show that both the cells exposed to NIR laser without TiO2 NTs treatment and the cells treated with TiO2 NTs but not with NIR irradiation had cell viabilities higher than 96%. Combination of these two techniques, however, shows cell viability less than 1%. The cell death rate strongly depended on the concentration of TiO2 NTs. Also, the cell deaths were mostly due to necrosis but partly due to late apoptosis. The in vivo animal test results show that tumor cells can be completely destroyed without nearly giving damage to surrounding healthy cells by an injection of an adequate amount of TiO2 NTs/NaCl suspension and a subsequent single continuous laser treatment at a moderately low laser illumina-tion intensity for the exposure time optimized for the tumor size. These results suggest that TiO2 NTs can be effectively utilized as a therapeutic agent for cancer thermotherapy due to their excellent photothermal property and high bio-compatibility.

New developments in the treatment of cutaneous leishmaniasis

Leishmaniasis,including the cutaneous form,poses an important public health threat around the world,while no vaccine is currently available against any form of leishmaniasis.The drugs used in the first line treatment of cutaneous leishmaniasis(CL)are commonly pentavalent antimonials despite their toxicities,long-term treatment duration and increasing resistance rates.Other alternatives are amphotericin B,pentamidine,miltefosine and paromomycine.Movement of the population,especially in endemic regions,increases the spread of the parasite and affectes the distribution of causative species,which requires re-evaluation the treatment regimen.Extensive researches are carried out on the treatment of leishmaniasis.The immunotherapeutic and targeted therapeutic approaches,formulations of carrier-loaded active drugs,local thermotherapeutic applications,the combination of antileishmanial drugs/compounds,the use of new synthetic and natural products are promising therapeutic options in the future.Herein,the author reviews the potential treatment modalities of CL with a brief overview of current treatments in the light of ongoing studies around the world.

Recirculating chemohyperthermia as a treatment for non-muscle invasive bladder cancer:Current and future perspectives

About 75% of all bladder cancer diagnosed are non-muscle invasive bladder cancer(NMIBC), recurring over 50% of them after transurethral resection of the bladder tumor. In order to prevent recurrences, adjuvant intravesical chemotherapy with mitomycin C and immunotherapy with bacillus Calmette-Gu-érin(BCG) is traditionally used. Unfortunately, many patients relapse after receiving these treatments and a significant proportion of them require surgery. After a one-to-three years BCG maintenance, the risk for progression at 5 years was 19.3% for T1G3 tumors. Many new treatment approaches are being investigated to increase the effectiveness of adjuvant intravesical therapy. One of the developing treatments for intermediate and high-risk NMIBC is the combination of intravesical chemotherapy and hyperthermia, called chemohyperthermia. This article provides a review of the mechanism of action, current status and indications, results and future perspectives.

Progress in cancer therapy with functionalized Fe_(3)O_(4) nanomaterials

Malignant neoplasms represent a significant global health threat.To address the need for accurate diagnosis and effective treatment,research is underway to develop therapeutic nanoplatforms.Iron oxide nanoparticles(NPs),specifically Fe_(3)O_(4) NPs have been extensively studied as potential therapeutic agents for cancer due to their unique properties including magnetic targeting,favorable biocompatibility,high magnetic response sensitivity,prolonged in vivo circulation time,stable performance,and high self-metabolism.Their ability to be integrated with magnetic hyperthermia,photodynamic therapy,and photothermal therapy has resulted in the widespread use of Fe_(3)O_(4) NPs in cancer diagnosis and treatment,making them a popular choice for such applications.Various methods can be employed to synthesize magnetic Fe_(3)O_(4) NPs,which can then be surface-modified with biocompatible materials or active targeting molecules.Multifunctional systems can be created by combining Fe_(3)O_(4) NPs with polymers.By combining various therapeutic approaches,more effective biomedical materials can be developed.This paper discusses the synthesis of Fe_(3)O_(4) NPs and the latest research advances in Fe_(3)O_(4)-based nanotherapeutic platforms,as well as their applications in the biomedical field.

Chemical chaperone delivered nanoscale metal–organic frameworks as inhibitor of endoplasmic reticulum for enhanced sensitization of thermo-chemo therapy

Thermotherapy and chemotherapy have received extensive attention to tumor treatment. However, thermal tolerance and drug resistance severely limit clinical effect of tumor therapy owing to endoplasmic reticulum(ER) stress. Reducing thermal tolerance and drug resistance of tumors is an urgent challenge to be solved. In this work, we design a nanoplatform of PBA-Dtxl@MIL-101 as an ER inhibitor. Amino functionalized Fe-metal organic framework(MIL-101) nanoparticles are synthesized as p H and microwave(MW) dual stimuli-responsive drug delivery system. Then, the chemical chaperones of 4-phenylbutyric acid(PBA) and antineoplastic drug Docetaxel(Dtxl) were successfully loaded into MIL-101 nanoparticles to form PBA-Dtxl@MIL-101 nanoparticles. Furthermore, PBA-Dtxl@MIL-101 nanoparticles exhibit inhibitor effect of ER stress through upregulating caspase 9 proteins and reduce thermal tolerance by downregulating HSP 90. It was demonstrated that the therapy sensitized by PBA-Dtxl@MIL-101 nanoparticles obviously destroyed tumor cells, showing simultaneously enhanced thermo-chemo therapy.