OBJECTIVE: To entrap carvacrol (CAR) in bovine serum albumin nanoparticles (BSANPs) to form CAR-loaded BSANPs (CAR@BSANPs) and to explore the anti-cancer effects in breast adenocarcinoma cells (MCF-7 cells) treated with CAR and CAR@BSANPs. METHODS: A desolvation method was used to synthesize BSANPs and CAR@BSANPs. The BSANPs and CAR@BSANPs were characterized by several physicochemical methods, including visual observation, high-resolution field emission scanning electron microscopy, Fourier transform infrared spectroscopy, and high-performance liquid chromatography. MCF-7 cells were used and analyzed after 24 h of exposure to CAR and CAR@BSANPs at half-maximal inhibitory concentration. The anti-proliferative, apoptotic, reactive oxygen species (ROS), and nitric oxide (NO) scavenging activity as well as gene expression analysis were investigated by the cell viability assay, phase-contrast microscopy, 2',7'-dichlorofluorescein-diacetate assay, Griess-Illosvoy colorimetric assay, and quantitative real-time polymerase chain reaction, respectively. RESULTS: CAR and CAR@BSANPs showed anti-proliferative, apoptotic, ROS generation, and NO scavenging effects on MCF-7 cells. Expression profile of B-cell lymphoma 2-like 11 (BCL2L11), vascular endothelial growth factor A (VEGFA), hypoxia inducible factor factor-1α (HIF1A), BCL2L11/apoptosis regulator (BAX), and BCL2L11/Bcl2 homologous antagonist/killer 1 (BAK1) ratios revealed downregulated genes; and BAX, BAK1, and CASP8 were upregulated by CAR and CAR@BSANPs treatment. In vitro anticancer assays of the CAR and CAR@BSANPs showed that CAR@BSANPs demonstrated higher therapeutic efficacy in the MCF-7 cells than CAR. CONCLUSIONS CAR and CAR@BSANPs affect gene expression and may subsequently reduce the growth and proliferation of the MCF-7 cells. Molecular targeting of regulatory genes of the MCF-7 cells with CAR and CAR@BSANPs may be an effective therapeutic strategy against breast cancer.
Humans ; Cymenes ; Nanoparticles/ultrastructure* ; MCF-7 Cells ; Breast Neoplasms/genetics* ; Apoptosis/drug effects* ; Serum Albumin, Bovine/chemistry* ; Monoterpenes/therapeutic use* ; Adenocarcinoma/genetics* ; Cell Proliferation/drug effects* ; Reactive Oxygen Species/metabolism* ; Female ; Cell Survival/drug effects* ; Animals ; Gene Expression Regulation, Neoplastic/drug effects* ; Nitric Oxide/metabolism* ; Cattle

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*

Huangqin Decoction(HQD) is a classic prescription for treating dysentery in the Treatise on Cold Damage and now is mainly used for the treatment of ulcerative colitis(UC). Since there are no requirements on specific Paeonia species, both Paeoniae Radix Alba(white peony root, WPR) and Paeoniae Radix Rubra(red peony root, RPR) are clinically used in HQD now. Although the two types of peony roots are close in origin and similar in primary components, the medicinal properties and efficacies are different. Furthermore, the systematic comparative analysis on the efficacy differences in treating UC of HQD with the roots of multi-originated peony has been seldom reported. This study compared and evaluated the pharmacological effects of HQD prepared from the roots of multi-originated peony, including WPR, RPR-l(derived from P. lactiflora), and RPR-v(derived from P. veitchii) based on the mouse model of UC induced by dextran sodium sulfate(DSS) by animal behaviors, pathological section(colon), and cytokine expression(IL-1β and IL-6), aiming to provide evidence for the identification of the original resource of peony root in HQD. The results indicated that all HQD samples prepared from WPR, RPR-l, and RPR-v could improve the symptoms of UC. Compared with the HQD-WPR, HQD-RPR-l and HQD-RPR-v were significantly different in weight loss, colon length, and disease activity index(DAI) score, but there was no significant difference between HQD-RPR-l and HQD-RPR-v. Moreover, HQD-RPR-v exhibited the most significant improvement in the pathological morphology of colonic tissue and mucosal defects. According to the previous comparative analysis of chemical profiling and content distribution of HQD prepared from the roots of multi-originated peony, RPR-v in HQD was potent in protecting against UC, which was presumedly attributed to a large number of monoterpene glycosides and galloyl glucoses. This study provided a scientific basis for the determination of peony root in HQD and its clinical medication.
Animals ; Colitis, Ulcerative/drug therapy* ; Dextran Sulfate ; Disease Models, Animal ; Drugs, Chinese Herbal/therapeutic use* ; Mice ; Monoterpenes ; Paeonia/chemistry* ; Plant Roots/chemistry*

OBJECTIVETo investigate the pro-angiogenic effects of paeoniflorin (PF) in a vascular insufficiency model of zebrafish and in human umbilical vein endothelial cells (HUVECs).

METHODSIn vivo, the pro-angiogenic effects of PF were tested in a vascular insufficiency model in the Tg(fli-1:EGFP)y1 transgenic zebrafish. The 24 h post fertilization (hpf) embryos were pretreated with vascular endothelial growth factor (VEGF) receptor tyrosine kinase inhibitor II (VRI) for 3 h to establish the vascular insufficiency model and then post-treated with PF for 24 h. The formation of intersegmental vessels (ISVs) was observed with a fluorescence microscope. The mRNA expression of fms-like tyrosine kinase-1 (flt-1), kinase insert domain receptor (kdr), kinase insert domain receptor like (kdrl) and von Willebrand factor (vWF) were analyzed by real-time polymerase chain reaction (PCR). In vitro, the pro-angiogenic effects of PF were observed in HUVECs in which cell proliferation, migration and tube formation were assessed.

RESULTSPF (6.25-100 μmol/L) could rescue VRI-induced blood vessel loss in zebrafish and PF (25-100 μmol/L), thereby restoring the mRNA expressions of flt-1, kdr, kdrl and vWF, which were down-regulated by VRI treatment. In addition, PF (0.001-0.03 μmol/L) could promote the proliferation of HUVECs while PF stimulated HUVECs migration at 1.0-10 μmol/L and tube formation at 0.3 μmol/L.

CONCLUSIONPF could promote angiogenesis in a vascular insufficiency model of zebrafish in vivo and in HUVECs in vitro.

Angiogenesis Inducing Agents ; pharmacology ; therapeutic use ; Animals ; Animals, Genetically Modified ; Cells, Cultured ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Embryo, Nonmammalian ; Glucosides ; pharmacology ; therapeutic use ; Human Umbilical Vein Endothelial Cells ; drug effects ; physiology ; Humans ; Monoterpenes ; pharmacology ; therapeutic use ; Neovascularization, Physiologic ; drug effects ; Phytotherapy ; Vascular Diseases ; drug therapy ; pathology ; Zebrafish

OBJECTIVE: To observe the efficacy of allergen blocking nasal spray (ABNS) , combined with seawater spray (SS) and myrtol standardized capsules (MSS), in patients with rhinitis sicca. METHOD: Patients diagnosed as rhinitis sicca were randomly divided into 2 groups: ABNS+SS group and SS group. In ABNS+SS group, patients used ABNS and SS four times per day and MSS twice per day. In SS group, patients used the same medicine except ABNS. Both group were observed for 14 days. The visual analog scale (VAS) of dryness, the endoscopy evaluation score (EES) and the tolerability of the spray were compared between the 2 groups. RESULT: There were 30 patients included in each group. The VAS and EES were improved in both groups after 2 weeks' treatment. The improvement of VAS and EES in ABNS+SS group was significantly superior to SS group (VAS: P < 0.01, EES: P < 0.05). There were only 2 patients complaint of slight discomfort when using ABNS, which did not disturb the treatment. CONCLUSION There is good efficacy of allergen blocking nasal spray (ABNS), combined with seawater spray (SS) and myrtol standardized capsules (MSS), in patients with rhinitis sicca.
Allergens ; Chronic Disease ; Drug Combinations ; Endoscopy ; Humans ; Monoterpenes ; therapeutic use ; Nasal Sprays ; Rhinitis ; drug therapy ; Sodium Chloride

OBJECTIVETo investigate the clinical effect of decoction of invigorating Qi and clearing lung combined standardized myrtol on acute exacerbation of chronic obstructive pulmonary disease (AECOPD).

METHODNinety and eight patients with AECOPD patients were randomly divided into the treatment group and the control group, with 50 cases and 48 cases respectively. All the patients were given the conventional treatment. The control group was treated by standardized myrtol with 3 times a day, 300 mg each time taken orally. The treatment group was given decoction of invigorating Qi and clearing lung with 2 times a day, one dose per day taken orally, combined standardized myrtol (usage as above). After Two weeks, the scores of clinical symptom, blood gas analysis and pulmonary function were observed.

RESULTBoth FEV1 and FEV1% were raised in the two groups after treating. And the treatment group was significantly higher than control group (P < 0.01). PaO2 and PaO2/FiO2 rose, with PaCO2 decreased in the two groups (P < 0.01). PaO2 and PaO2/FiO2 were significantly improved, and PaCO2 was significantly decreased in the treatment group compared to the control group (P < 0.01). In the clinical curative effect comparison aspects, clinical control rates were 42.0% in treatment group and 20.83% in control group respectively, with significant difference between the two groups (P < 0.05). Significant efficiency is 86.0% in treatment group and 52.08% in control group respectively, with significant difference between the two groups (P < 0.01).

CONCLUSIONDecoction of invigorating Qi and clearing lung combined with standardized myrtol can obviously improve clinical symptom, blood gas an analysis and pulmonary function in patients with AECOPD.

Aged ; Blood Gas Analysis ; Drug Combinations ; Drug Therapy, Combination ; Female ; Humans ; Lung ; physiopathology ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Monoterpenes ; standards ; therapeutic use ; Pulmonary Disease, Chronic Obstructive ; blood ; physiopathology ; therapy ; Reference Standards ; Respiratory Function Tests ; Treatment Outcome

OBJECTIVE: To observed the prevention efficacy of secretory otitis media after radiation therapy by the Myrtol Standardized Enteric Coated Soft Capsules. METHOD: Sixty patients with nasopharyngeal carcinoma who Diagnosis without secretory otitis media before radiation therapy were divided into experimental group and control group, 30 cases in each group. After the start of radiation therapy ,the experimental group patients oral the Myrtol Standardized Enteric Coated Soft Capsules, each 0.3 g, 3 times a day, 7 days a course of treatment, oral the medication three months, the patients in the control group received no treatment. 3 months and 6 months after the end of radiation therapy, whether there is a difference comparison of experimental group and the control group in symptoms, signs, pure tone audiometry and tympanogram change. RESULT: Seventeen patients (18 ears) (56.67%, 17/30) in the control group were suffering from secretory otitis media, 7 patients (7 ears) (23.33%, 7/30) in the experimental group were suffering from secretory otitis media. The difference between the two groups was statistically significant (P < 0.01). 17 patients (17 ears) in the control group and 7 patients (7 ears) in the experimental group were suffering from tinnitus. 20 patients(20 ears) in the control group and 9 patients (10 ears) in the experimental group have ear choking feeling. The difference between the two groups was statistically significant (P < 0.01). The air conduction hearing threshold of the experimental group before radiation therapy is (7.5 +/- 2.0) dB HL and the air conduction hearing threshold of the control group patients is (8.3 +/- 4.0) dB HL. The difference between the two groups was not statistically significant (P > 0.05). 3 months after radiation therapy,the gas conductive hearing threshold of the experimental group is (25.6 +/- 3.0) dB HL, but the data in the control group is (40.7 +/- 5.0) dB HL. The difference between the two groups was statistically significant (P < 0.01). CONCLUSION Patients with nasopharyngeal carcinoma oral the the Myrtol Standardized Enteric Coated Soft Capsules before radiation therapy can effectively reduce the incidence of secretory otitis media after radiotherapy, it can prevent the occurrence of secretory otitis media.
Adult ; Aged ; Aged, 80 and over ; Carcinoma ; Drug Combinations ; Female ; Humans ; Male ; Middle Aged ; Monoterpenes ; therapeutic use ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; radiotherapy ; Otitis Media with Effusion ; etiology ; prevention & control ; Radiotherapy ; adverse effects

OBJECTIVE: To explore the effect of mucoregulatory agents during endoscopic sinus surgery. METHOD: Ninety-seven cases with chronic rhinosinusitis were randomly divided into three groups, with 31 cases in B group treated by ambroxol, 33 cases in C group treated by eucalyptol-limonene-pinene enteric soft capsule and 33 cases in control group (A group). The follow-up visit lasted for 6 months for three groups. Then, the therapeutic effects were evaluated and compared among these three groups. RESULT: By the end of 6 months after treatment,the effective rate was 90.3% and 97.0% for cases in the B and C groups respectively, and only 75.8% in A group. The difference between A and C was statistically significant (P < 0.05). CONCLUSION Eucalyptol-limonene-pinene enteric soft capsule,as a multicomponent mucoregulatory agent, can obviously improve the secretion of mucosa and epithelial recovery, thus accelerate healing of the disease. It can also improve the success rate of functional endoscope sinus surgery, and may play a promising role in clinical application.
Adolescent ; Adult ; Aged ; Ambroxol ; therapeutic use ; Chronic Disease ; Cyclohexanols ; therapeutic use ; Cyclohexenes ; therapeutic use ; Eucalyptol ; Expectorants ; therapeutic use ; Female ; Humans ; Intraoperative Period ; Limonene ; Male ; Middle Aged ; Monoterpenes ; therapeutic use ; Sinusitis ; drug therapy ; Terpenes ; therapeutic use ; Treatment Outcome ; Young Adult

OBJECTIVETo study the influence of Chinese herbs for inner-warming on the bioavailability of paeoniflorin (PF) and its mechanism.

METHODSChinese herbs (pepper fruit, evodia fruit, cassia bark, fennel fruit and prickly-ash peel) were separately used in combination with PF for gastrogavage to mice. Reversed phase high-performance liquid chromatography was used to determine the plasma concentration of PF in mice after medication. The bioavailability of PF was calculated and compared, taking single use of red peony root for control.

RESULTSThe pharmacokinetics of PF in mice was conformed to the one-compartment model, as combined use with Chinese herbs for inner-warming, the relative bioavailability of PF was 137.22% for pepper fruit, 123.62% for evodia fruit, 108.39% for cassia bark, 226.02% for fennel fruit and 116.73% for prickly-ash peel, there were difference of Cmax and AUC(0-infinity) in comparison of these data with the control group (P < 0.05), but with no difference of tmax (P > 0.05).

CONCLUSIONThe Chinese herbs used in this experiment in combination with red peony root could enhance the bioavailability of PF, which illustrated the scientific meaning of the recipe combination of Chinese herbs for activating blood circulation and inner-warming viewing from pharmacodynamics.

Animals ; Benzoates ; administration & dosage ; isolation & purification ; pharmacokinetics ; Biological Availability ; Bridged-Ring Compounds ; administration & dosage ; isolation & purification ; pharmacokinetics ; Cassia ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacokinetics ; pharmacology ; therapeutic use ; Evodia ; chemistry ; Female ; Glucosides ; administration & dosage ; isolation & purification ; pharmacokinetics ; Male ; Mice ; Monoterpenes ; Paeonia ; chemistry ; Phytotherapy ; Random Allocation