OBJECTIVE: "Multi-targeting" drugs can prove fruitful to combat drug-resistance of multifactorial disease-cervical cancer. This study envisioned to reveal if Thuja homeopathic mother tincture (MT) and its bioactive component could combat human papillomavirus (HPV)-16-infected SiHa cervical cancer cells since it is globally acclaimed for HPV-mediated warts. METHODS: Thuja MT was studied for its antiproliferative and antimigratory properties in SiHa cells followed by microscopic determination of reactive oxygen species (ROS) generation by 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) staining and loss in mitochondrial membrane potential (MtMP) by rhodamine 123 (Rh123) staining. Apoptosis and autophagy inductions were studied by acridine orange/ethidium bromide (AO/EB) staining and immunoblot analyses of marker proteins. The bioactive component of Thuja MT detected by gas chromatography-mass spectrometry was studied for antiproliferative and antimigratory properties along with in silico prediction of its cellular targets by molecular docking and oral drug forming competency. RESULTS: Thuja MT showed significant antiproliferative and antimigratory potential in SiHa cells at a 50% inhibitory concentration (IC₅₀) of 17.3 µL/mL. An increase in DCFDA fluorescence and loss in Rh123 fluorescence prove that Thuja MT acted through the burst of ROS and loss in MtMP respectively. AO/EB-stained cells under the microscope and immunoblot analyses supported Thuja-induced cellular demise via dual pathways-apoptosis and autophagy. Immunoblots showed cleavage of caspase-3 and poly(adenosine diphosphate-ribose) polymerase-1 (PARP-1) along with upregulation of Beclin-1, microtubule-associated protein 1 light chain 3B (LC3B)-II, and p62 proteins. Hence, the apoptotic cascade followed a caspase-3-dependent pathway supported by PARP-1 cleavage, while autophagic death was Beclin-1-dependent and mediated by accumulation of LC3BII and p62 proteins. Thujone, detected as the bioactive principle of Thuja MT, showed greater anti-proliferative and anti-migratory potential at an IC₅₀ of 77 µg/mL, along with excellent oral drug competency with the ability for gastrointestinal absorption and blood-brain-barrier permeation with nil toxicity. Molecular docking depicted thujone with the strongest affinity for mammalian target of rapamycin, phosphoinositide 3-kinase, and protein kinase B followed by B-cell lymphoma 2, murine double minute 2 and adenosine monophosphate-activated protein kinase, which might act as upstream triggers of apoptotic-autophagic crosstalk. CONCLUSION Robust "multi-targeting" anticancer potential of Thuja drug and thujone for HPV-infected cervical cancer ascertained its therapeutic efficacy for HPV infections.
Animals ; Apoptosis ; Autophagy ; Beclin-1/pharmacology* ; Bicyclic Monoterpenes ; Caspase 3 ; Cell Line, Tumor ; Female ; Humans ; Mammals/metabolism* ; Mice ; Molecular Docking Simulation ; Papillomavirus Infections/drug therapy* ; Phosphatidylinositol 3-Kinases ; Poly(ADP-ribose) Polymerase Inhibitors/therapeutic use* ; Reactive Oxygen Species/metabolism* ; Thuja/metabolism* ; Uterine Cervical Neoplasms/pathology*

Objective: To investigate the sensitization characteristics of Juniperus chinensis pollen in patients with allergic rhinitis and/or allergic asthma in Beijing area, and to explore the characteristics of Juniper chinensis pollen sensitized population. Methods: Patients with suspected allergic rhinitis and/or asthma from January 2017 to December 2019 in the outpatient department of Allergy Department of Beijing Shijitan Hospital were selected in this study. Skin prick test (SPT) was performed with Juniper chinensis pollen allergen reagent to compare different age and disease allergen distribution, and to observe the sensitization characteristics of its population. All of the analyses were performed using SAS software version 9.4. Results: A total of 8 380 patients were enrolled in the end. The total positive rate of Juniper chinensis pollen SPT reached 49.92% (4 183/8 380). The positive rate of Juniper chinensis pollen SPT was highest in the 10-14 age group, reaching 60.99% (283/464). Compared with other age groups, there was a statistical difference (χ²=266.77, P<0.01). The SPT positive rate of patients aged less than 10 years increased with the increase of age, while the SPT positive rate of patients aged over 40 years decreased with the increase of age. Single Juniper chinensis pollen was less allergenic, accounting for about 25.05% (1 048/4 183), and the patients' age was (35.21±12.39) years. Regardless of single Juniper chinensis pollen or other pollen allergies, allergic rhinitis was the main disease. Among the patients with SPT positive Juniper chinensis pollen combined with other inhaled pollen allergens, willow pollen accounted for the first (74.99%). The positive rate of Juniper chinensis pollen was the highest in patients with single allergic rhinitis, accounting for 52.05% (3 797/7 295), and the rate in patients with single allergic asthma was the lowest, accounting for 17.49% (53/303), with statistically difference (χ²=138.99, P<0.01). Conclusions: Juniper chinensis pollen is highly sensitized in patients with allergic rhinitis and/or allergic asthma in Beijing . The positive rate of SPT is highest among 10-14 age group, most of which showed strong positive reaction, and allergic rhinitis is more common in Juniper chinensis pollen sensitization diseases.
Adolescent ; Adult ; Allergens ; Asthma ; Child ; Humans ; Juniperus ; Pollen ; Rhinitis, Allergic ; Skin Tests

A pollen/food-associated syndrome (PFAS) has been described between peach and cypress pollen. Cross-reactive allergens were characterized which belong to the Gibberellin-regulated protein (GRP) family, BP14 in cypress pollen and Pru p 7 in peach. GRP are small cationic protein with anti-microbial properties. A patient suffering from a peach/cypress syndrome was explored clinically and biologically using 2 types of immunoglobulin E (IgE) multiarray microchip, immunoblots and a basophil activation test to assess the clinical relevance of various extracts and purified allergens from fruits or cypress pollen. In addition to PR10 sensitization, the patient showed specific IgE to Pru p 7, BP14 and allergen from pomegranate. These last 3 allergens and allergenic sources are able to induce ex vivo basophil activation characterized by the monitoring of the expression of CD63 and CD203c, both cell surface markers correlated with a basophil mediator release. Up to 100% of cells expressed CD203c at 50 ng/mL of BP14 protein. In contrast, snakin-1, a GRP from potato sharing 82% sequence identity with Pru p 7 did not activate patient's basophils. These results strongly suggest that, like Pru p 7, BP14 is a clinically relevant allergenic GRP from pollen. Allergen members of this newly described protein family are good candidates for PFAS where no cross-reactive allergens have been characterized.
Allergens ; Basophils ; Cupressus* ; Fruit ; Humans ; Immunoglobulin E ; Immunoglobulins ; Pollen* ; Prunus persica ; Punicaceae ; Solanum tuberosum

OBJECTIVES: Tree pollen causes allergic rhinitis and asthma. We investigated children who diagnosed as rhinitis or asthma, living in Busan, for tree allergen sensitization, component allergen, oral allergy syndrome, and the relationship between pollen counts and allergen sensitization.METHODS: Pollen were collected in Busan, from January 1 to December 31, 2017, using a Rotorod sampler and enumerated using a microscope. We conducted a study of children with rhinitis or asthma at Busan St. Mary's Hospital in 2017, administered an ISAAC questionnaire, and an oral allergy syndrome survey. Serum specific Ig E tests were performed.RESULTS: Among the 57 patients, the mean age was 9.3 years. The pollen counts in decreasing order were as follows: pine, alder, oak, juniper, beech, ginkgo, and birch. For sensitization, birch and alder 35.1%, Japanese cedar 19.3%, juniper 17.5%, pine 10.5%, and Japanese cypress 8.2%. The component Ig E was tested in 27 patients. Bet v 1 had a high correlation with birch, alder, and peach. Bet v 2 showed a statistically significant correlation with all tree pollen except cypress. Bet v 4 did not have any apparent correlation. Bet v 6 had the same pattern as Bet v 2, but correlation coefficient was higher than that of Bet v 2. Oral allergy syndrome was noted in 7 patients, including peach, peanut, apple, tomato, kiwi, and sesame.CONCLUSIONS: Alder and juniper are clinically important tree pollens in Busan. These pollens cause sensitization to birch and Japanese cedar by cross-reaction.
Alnus ; Arachis ; Asian Continental Ancestry Group ; Asthma ; Betula ; Busan ; Child ; Cryptomeria ; Cupressus ; Fagus ; Ginkgo biloba ; Humans ; Hypersensitivity ; Juniperus ; Lycopersicon esculentum ; Pollen ; Prunus persica ; Rhinitis ; Rhinitis, Allergic ; Sesamum ; Trees

Berries and branches essential oil of Juniperus phoenicea were obtained by electromagnetic induction heating assisted extraction and by hydrodistillation with a yield varied from (1.2 ± 0.3 to 2.4 ± 0.7%) and from (0.6 ± 0.1% to 1.1 ± 0.1%), respectively. forty eight compounds were identified representing (97.2 – 99.7%) of the oil. α-Pinene (40.3 – 67.8%) and δ-3-carene (13.5 – 26.8%) were the main compounds in berries and branches essential oils. Antioxidant activity was evaluated by three means: inhibition of 2, 2-diphenyl-1-picryl hydrazyl (DPPH) free radical, reducing power and β-Carotene/linoleic acid bleaching. The antioxidant activity of essential oils showed IC₅₀ ranging from 67.6 ± 1.02 µg/mL to 131.5 ± 0.8 µg/mL for berries and from 98 ± 1.25 µg/mL to 166.8 ± 0.29 µg/mL for the branches. Berries oil show more potent antioxidant activity compared to branches. This result is supported by the three methods investigated in this work.
Fruit ; Heating ; Hot Temperature ; Juniperus ; Magnets ; Oils, Volatile

OBJECTIVEThis study examined the antimicrobial activity of Cannabis sativa, Thuja orientalis and Psidium guajava against methicillin-resistant Staphylococcus aureus (MRSA) and used a standardized purification protocol to determine the presence and abundance of bioactive compounds in the leaf extracts.

METHODSIn vitro antimicrobial activities of the ethanolic extracts of C. sativa, T. orientalis and P. guajava were tested against MRSA. The presence of bioactive molecules in these three leaves was evaluated using biochemical assays and high-performance thin-layer chromatography (HPTLC).

RESULTSResistance to methicillin, penicillin, oxacillin and cefoxitin was observed in each of the clinical and nonclinical MRSA isolates. However, they were still vulnerable to vancomycin. Used individually, the 50% extract of each plant leaf inhibited MRSA growth. A profound synergism was observed when C. sativa was used in combination with T. orientalis (1:1) and when P. guajava was used in combination with T. orientalis (1:1). This was shown by larger zones of inhibition. This synergism was probably due to the combined inhibitory effect of phenolics present in the leaf extracts (i.e., quercetin and gallic acid) and catechin, as detected by HPTLC.

CONCLUSIONThe leaf extracts of C. sativa, T. orientalis and P. guajava had potential for the control of both hospital- and community-acquired MRSA. Moreover, the inhibitory effect was enhanced when extracts were used in combination.

Anti-Bacterial Agents ; pharmacology ; Cannabis ; Drug Resistance ; drug effects ; Humans ; Methicillin ; pharmacology ; Methicillin-Resistant Staphylococcus aureus ; drug effects ; growth & development ; Microbial Sensitivity Tests ; Phytotherapy ; Plant Extracts ; pharmacology ; Plant Leaves ; Psidium ; Staphylococcal Infections ; drug therapy ; microbiology ; Thuja

A new sesquiterpenoid, 11-hydroxy-valenc-1(10),3(4)-dien-2-one (3), two chemically synthesized but first isolate from nature, 3-oxocedran-8β-ol (1) and valenc-1(10),3(4),11(12)-trien-2-one (2) along with four known compounds, sugiol (4), (+)-nootkatone (5), 11-hydroxy-valenc-1(10)-en-2-one (6), and clovandiol (7), were isolated from the heartwood of Juniperus chinensis. All chemical structures were elucidated using extensive spectroscopic analysis including 1D and 2D NMR spectroscopy. Valenc-1(10),3(4),11(12)-trien-2-one (2) exhibited significant inhibitory activity against butyrylcholinesterase with an IC₅₀ value of 68.45 µM.
Acetylcholinesterase ; Butyrylcholinesterase ; Juniperus ; Magnetic Resonance Spectroscopy

Omalizumab is frequently used as add-on treatment to inhaled corticosteroids (ICS) and long-acting β2-agonists in patients with suboptimal control of severe asthma. Patients with severe asthma will typically require chronic treatment, although due to the limited amount of data available there are still some concerns about the safety and efficacy of long-term therapy with omalizumab. Herein, in an extension of a previous 4-year study, we report disease-related outcomes of 8 patients with severe persistent allergic asthma who have been followed for a total of 9 years in a real-life setting. Both quality of life (QoL) (evaluated using the Juniper Asthma-Related QoL Questionnaire [AQLQ]) and forced expiratory volume in 1 second (FEV1) showed sustained improvement at 9 years. The median values of AQLQ and FEV1 at 4 years were 5.5 and 82.0% compared to 5.9 and 85.5%, respectively, at 9 years, which were all significantly increased from baseline. After 9 years, the mean annual number of severe exacerbations was 0.63 compared to 5 at baseline. There also appeared to be a trend toward use of a lower dose of ICS at longer follow-up times. After 9 years, there were no safety concerns for continued use of omalizumab, and no asthma-related hospitalizations or emergency department visits were documented over the last 5 years. The present analysis is the longest reported clinical follow-up of omalizumab. Long-term maintenance treatment with omalizumab for up to 9 years is associated with continued benefits in reducing symptoms, exacerbations, and medication burden without any safety concerns.
Adrenal Cortex Hormones ; Asthma ; Emergency Service, Hospital ; Follow-Up Studies* ; Forced Expiratory Volume ; Hospitalization ; Humans ; Juniperus ; Omalizumab* ; Quality of Life

BACKGROUND: Specific IgE antibodies against the low-molecular-weight carbohydrate antigen that does not bridge IgE molecules on mast cells are not associated with clinical symptoms. Cross reactivity can be determined in allergen-specific IgE detection assays when the carbohydrate structures between pollen allergens and plant derived food allergens are similar; in such cases, false positive results for grain or legume allergens can be reported for pollen allergic patients who are not sensitized to those allergens. This phenomenon arises owing to the presence of cross-reactive carbohydrate determinants (CCDs). OBJECTIVE: This study aimed to assess the impact of CCD interference on the results for pollen allergen-specific IgE antibodies in the general adult population and to perform CCD inhibition tests evaluating the involvement of CCD on samples positive to pollen allergens. METHODS: Serum samples from 322 subjects were tested for IgE antibodies to pollens and CCD. The research subjects were given questionnaires about pollen allergic symptoms to help assess the presence of allergies. Allergen IgE antibodies for Japanese cedar, Japanese cypress, orchard grass, ragweed, MUXF, bromelain, horseradish peroxidase (HRP), and ascorbate oxidase (ASOD) were analyzed. RESULTS: It was observed that among individuals who tested positive to any of the pollen allergens, the positive ratio of CCD-specific IgE antibody was the highest for HRP (13.5%–50.0%). The results from the inhibition tests revealed that CCD was marginally present. Although IgE antibodies for cedar pollen did not react with CCD, IgE antibodies for Japanese cypress, orchard grass, and ragweed might be detected by the presence of CCD. CONCLUSION: The results of the inhibition tests revealed the obvious presence of CCD suggesting its involvement. Considering these findings, careful evaluation of patient IgE results should be performed for Japanese cypress, orchard grass, and ragweed.
Adult ; Allergens ; Ambrosia ; Antibodies ; Ascorbate Oxidase ; Asian Continental Ancestry Group ; Bromelains ; Cryptomeria ; Cupressus ; Dactylis ; Fabaceae ; False Positive Reactions ; Horseradish Peroxidase ; Humans ; Hypersensitivity ; Immunoglobulin E ; Mast Cells ; Plants ; Pollen ; Research Subjects ; Rhinitis, Allergic ; Rhinitis, Allergic, Seasonal

BACKGROUND AND OBJECTIVES: The biological activity in antibacterial and antioxidative action of essential oils (EOs) have been investigated. In this study, we tried to evaluate the effects of Chamaecyparis obtusa EOs on producing chemical mediators by peripheral blood mononuclear cells (PBMCs). SUBJECTS AND METHOD: PBMCs from healthy volunteers were stimulated with lipopolysaccharide (LPS) and phytohaemagglutinin (PHA) in the presence of varying concentrations of EOs. Cytotoxic effects of EOs were measured using an aqueous cell proliferation assay kit and supernatants were analyzed using enzyme-linked immunosorbent assay. Tumor necrosis factor-α (TNF-α), interleukin-5, and interferon-γ (INF-γ) protein levels were measured to determine the anti-inflammatory effects of essential oil. RESULTS: EOs were found to have cytotoxic effects on PBMCs at levels of over 1%. EOs not only could induce PBMCs to produce chemical mediators, but it also significantly inhibited the LPS induced TNF-α and INF-γ productions as well as the PHA induced INF-γ production. CONCLUSION: EOs had cytotoxic effects at high concentrations and modulated chemical mediator productions from PBMC. These data suggest that EOs could be used to treat immunologic or inflammatory diseases.
Cell Proliferation ; Chamaecyparis* ; Enzyme-Linked Immunosorbent Assay ; Healthy Volunteers ; Interleukin-5 ; Methods ; Necrosis ; Oils, Volatile