Anticariogenic Activity of Celastrol and Its Enhancement of Streptococcal Antagonism in Multispecies Biofilm.

Dental caries is a chronic disease resulting from dysbiosis in the oral microbiome. Antagonism of commensal Streptococcus sanguinis and Streptococcus gordonii against cariogenic Streptococcus mutans is pivotal to keep the microecological balance. However, concerns are growing on antimicrobial agents in anticaries therapy, for broad spectrum antimicrobials may have a profound impact on the oral microbial community, especially on commensals. Here, we report celastrol, extracted from Traditional Chinese Medicine's Tripterygium wilfordii (TW) plant, as a promising anticaries candidate. Our results revealed that celastrol showed antibacterial and antibiofilm activity against cariogenic bacteria S. mutans while exhibiting low cytotoxicity. By using a multispecies biofilm formed by S. mutans UA159, S. sanguinis SK36, and S. gordonii DL1, we observed that even at relatively low concentrations, celastrol reduced S. mutans proportion and thereby inhibited lactic acid production as well as water-insoluble glucan formation. We found that celastrol thwarted S. mutans outgrowth through the activation of pyruvate oxidase (SpxB) and H2O2-dependent antagonism between commensal oral streptococci and S. mutans. Our data reveal new anticaries properties of celastrol that enhance oral streptococcal antagonism, which thwarts S. mutans outgrowth, indicating its potential to maintain oral microbial balance for prospective anticaries therapy.

Differences in medical costs between TCM users and TCM nonusers in inpatients with thalassemia.

BACKGROUND: Thalassemia has brought serious health threats and economic burdens to patients worldwide. There is no sovereign remedy for thalassemia, both conventional and Traditional Medicine (TM) methods have certain effects on this disease. As typical of TM, Traditional Chinese Medicine (TCM) has been widely used in the treatment of thalassemia. Previous studies mainly focused on conventional treatments for thalassemia and patients' medical burden, but no research has examined the effects of TCM use on the economic burdens for thalassemia inpatients in mainland China. The main objective of this study is to compare the medical cost differences between TCM users and TCM nonusers, furtherly, we will discuss the role of TCM use in the treatment of thalassemia. METHODS: We employed the 2010-2016 Medicare claims database provided by the China Health Insurance Research Association (CHIRA). Chi-square and Mann-Whitney tests were used to analyze the differences between TCM users and TCM nonusers. Multiple regression analysis was performed using the ordinary least squares method to compare the TCM users' inpatient medical cost with TCM nonusers', and to further examine the correlation between TCM cost, conventional medication cost and nonpharmacy cost for TCM users. RESULTS: A total of 588 urban thalassemia inpatients were identified, including 222 TCM users and 366 TCM nonusers. The inpatient medical cost of TCM users was RMB10,048 (USD1,513), which was significantly higher than TCM nonusers (RMB1,816 (USD273)). Total inpatient cost for TCM users was 67.4% higher than those of TCM nonusers (P < 0.001). With confounding factors fixed, we found that the conventional medication cost and nonpharmacy cost were positively correlated with TCM cost. CONCLUSION: Total hospitalization expenses for TCM users were higher than TCM nonusers. Both the conventional medication cost and nonpharmacy cost of TCM users were all higher than TCM nonusers. We infer TCM plays a complementary role, rather than an alternative, in the treatment of thalassemia due to the lack of cooperative treatment guidelines. It is recommended that a cooperative diagnosis and treatment guidelines should be generated to balance the use of TCM and conventional medicine for treating thalassemia, so as to reduce the economic burdens on patients.

Costs of traditional Chinese medicine treatment for inpatients with lung cancer in China: a national study.

BACKGROUND: Traditional Chinese Medicine (TCM) has long been a widely recognized medical approach and has been covered by China's basic medical insurance schemes to treat lung cancer. But there was a lack of nationwide research to illustrate the impact of the use of TCM on lung cancer patients' economic burden in mainland China. Therefore, we conduct a nationwide study to reveal whether the use of TCM could increase or decrease the medical expenditure of lung cancer inpatients in mainland China. METHODS: This is a 7-year cross-sectional study from 2010 to 2016. The data is a random sample of 5% from lung cancer claims data records of Chinese Urban Employee Basic Medical Insurance (UEBMI) and Urban Resident Basic Medical Insurance (URBMI). Mann-Whitney test was used to compare inpatient cost data with positive skewness. Ordinary least squares regression analysis was performed to compare the total TCM users' hospitalization cost with TCM nonusers', to examine whether TCM use is the key factor inducing relatively high medical expenditure. RESULT: A total of 47,393 lung cancer inpatients were included in this study, with 38,697 (81.7%) of them at least using one kind of TCM approach. The per inpatient medical cost of TCM users was RMB18,798 (USD2,830), which was 65.2% significantly higher than that of TCM nonusers (P < 0.001). The medication cost, conventional medication cost, and nonpharmacy cost of TCM users were all higher than TCM nonusers, illustrating the higher medical cost of TCM users was not induced by TCM only. With confounding factors fixed, there was a positive correlation between TCM cost and conventional medication cost, nonpharmacy cost (Coef. = 0.283 and 0.211, all P < 0.001), indicting synchronous increase of TCM costs and conventional medication cost for TCM users. CONCLUSION: The use of TCM could not offset the utilization of conventional medicine, demonstrating TCM mainly played a complementary role but not an alternative role in the inpatient treatment of lung cancer. A joint Clinical Guideline that could balance the use of TCM and Conventional medicine should be developed for the purpose of reducing economic burden for lung cancer inpatients.

Baicalein exerts anxiolytic and antinociceptive effects in a mouse model of posttraumatic stress disorder: Involvement of the serotonergic system and spinal delta-opioid receptors.

Post-traumatic stress disorder (PTSD) is a serious mental disease featured by a stress dysfunction that occurs after an individual has faced intense mental stress, often accompanied by anxiety and chronic pain. Currently, the mainstream drug for PTSD is serotonin reuptake inhibitors (SSRIs), however, their pain management for patients is limited. Baicalein, a Chinese traditional herbal medicine, has shown promising results in treating anxiety, depression, and pain. In this study, we found that baicalein may alleviate single prolonged stress (SPS)-induced PTSD-like behaviors in mice without altering baseline nociceptive sensitivity or activity. Meanwhile, baicalein increased the noradrenaline (NE) and serotonin (5-HT) content and decreased the ratio of 5-hydroxyindoleacetic acid (5-HIAA)/5-HT by inhibiting the activity of monoamine oxidase A (MAO-A) in SPS-induce mice. The anxiolytic and antinociceptive effects induced by baicalein were totally abolished by 5-HT depleting agents. Moreover, the anxiolytic effects of baicalein could be abolished by the 5-HT1A receptor antagonist WAY-100635, and the analgesic effects could be abolished by delta-opioid receptor antagonists in the spinal. Taken together, our study provides compelling evidence that baicalein reversed anxiety-like behaviors and neuropathic pain in PTSD through serotonergic system and spinal delta-opioid receptors.

Use of Traditional Chinese Medicine and Its Impact on Medical Cost among Urban Ischemic Stroke Inpatients in China: A National Cross-Sectional Study.

BACKGROUND: Traditional Chinese medicine (TCM) has long been widely adopted by the Chinese people and has been covered by China's basic medical insurance schemes to treat ischemic stroke. Previous research has mainly highlighted the therapy effect of TCM on ischemic stroke patients. Some studies have demonstrated that employing TCM can reduce the medical burden on other diseases. But no research has explored whether using TCM could reduce inpatient medical cost for ischemic stroke in mainland China. The purpose of this study is to investigate the impact of the use of TCM on the total inpatient cost of ischemic stroke and to explore whether TCM has played the role of being complementary to, or an alternative for, conventional medicine to treat ischemic stroke. METHODS: We conducted a national cross-sectional analysis based on a 5% random sample from claims data of China Urban Employee Basic Medical Insurance (UEBMI) and Urban Resident Basic Medical Insurance (URBMI) schemes in 2015. Mann-Whitney test was used to compare unadjusted total inpatient cost, conventional medication cost, and nonpharmacy cost estimates. Ordinary least square regression analysis was performed to compare demographics-adjusted total inpatient cost and to examine the association between TCM cost and conventional medication cost. RESULTS: A total of 47321 urban inpatients diagnosed with ischemic stroke were identified in our study, with 92.6% (43843) of the patients using TCM in their inpatient treatment. Total inpatient cost for TCM users was significantly higher than TCM nonusers (USD 1217 versus USD 1036, P < 0.001). Conventional medication cost was significantly lower for TCM users (USD 335 versus USD 436, P < 0.001). The average cost of TCM per patient among TCM users was USD 289. Among TCM users, conventional medication costs were found to be positively associated with TCM cost after adjusting for confounding factors (Coef. = 0.144, P < 0.001). CONCLUSION: Although the use of TCM reduced the cost of conventional medicine compared with TCM nonusers, TCM imposed an extra financial component on the total inpatient cost on TCM users. Our study suggests that TCM mainly played a complementary role to conventional medicine in ischemic stroke treatment in mainland China.

Novel Inhalable Ciprofloxacin Dry Powders for Bronchiectasis Therapy: Mannitol-Silk Fibroin Binary Microparticles with High-Payload and Improved Aerosolized Properties.

Non-cystic fibrosis bronchiectasis (NCFB) is a chronic respiratory disease associated with the high morbidity and mortality. Long-term intermittent therapy by inhalable antibiotics has recently emerged as an effective approach for NCFB treatment. However, the effective delivery of antibiotics to the lung requires administering a high dose to the site of infection. Herein, we investigated the novel inhalable silk-based microparticles as a promising approach to deliver high-payload ciprofloxacin (CIP) for NCFB therapy. Silk fibroin (SF) was applied to improve drug-payload and deposit efficiency of the dry powder particles. Mannitol was added as a mucokinetic agent. The dry powder inhaler (DPI) formulations of CIP microparticles were evaluated in vitro in terms of the aerodynamic performance, particle size distribution, drug loading, morphology, and their solid state. The optimal formulation (highest drug loading, 80%) exhibited superior aerosolization performance in terms of fine particle fraction (45.04 ± 0.84%), emitted dose (98.10 ± 1.27%), mass median aerodynamic diameter (3.75 ± 0.03 µm), and geometric standard deviation (1.66 ± 0.10). The improved drug loading was due to the electrostatic interactions between the SF and CIP by adsorption, and the superior aerosolization efficiency would be largely attributed to the fluffy and porous cotton-like property and low-density structure of SF. The presented results indicated the novel inhalable silk-based DPI microparticles of CIP could provide a promising strategy for the treatment of NCFB.

Thermo-sensitive gel in glaucoma therapy for enhanced bioavailability: In vitro characterization, in vivo pharmacokinetics and pharmacodynamics study.

AIMS: Glaucoma is a chronic ophthalmic disease, which has become one of the leading causes to progressive and irreversible blindness. Current ophthalmic drug delivery to treat glaucoma is mostly eyedrop, whose rapid elimination on corneal surface can lead to poor bioavailability. The present study was aimed to develop a timolol maleate loaded thermo-sensitive gel (TM-TSG) with improved bioavailability to treat glaucoma. MAIN METHODS: TM-TSG was prepared by homogeneously dispersing 0.3% (w/v) timolol maleate, 24.25% (w/v) poloxamer 407 (P407) and 1.56% (w/v) poloxamer 188 (P188) into phosphate buffer solution (pH = 7.4) and the formulated TM-TSG was characterized. KEY FINDINGS: TM-TSG was stored in liquid form at room temperature (25 °C) and transited to semisolid gel at physiological temperature (32 °C). The rheological property of TM-TSG was in favor of uniform distribution of drug. TM-TSG showed good stability at different conditions including centrifugation, autoclaving and different temperature. In vivo pharmacokinetic studies indicated that TM-TSG could enhance absorption of TM in aqueous humor and improve the ocular bioavailability in comparison of commercial TM eyedrops. In vivo experiment result showed that TM-TSG had greater effect in treating glaucoma than TM eyedrops by sustainably lowering intraocular pressure (IOP) for a week. Moreover, slit lamp test and histopathological analysis demonstrated that TM-TSG had excellent biocompatibility. SIGNIFICANCE: TM-TSG could be a promising ophthalmic delivery system for glaucoma therapy.

Anhydrous reverse micelle nanoparticles: new strategy to overcome sedimentation instability of peptide-containing pressurized metered-dose inhalers.

The objective of this study was to develop a novel anhydrous reverse micelle nanoparticles (ARM-NPs) system to overcome the sedimentation instability of peptide-containing pressurized metered-dose inhalers (pMDIs). A bottom-up method was utilized to fabricate ARM-NPs. Tertiary butyl alcohol (TBA)/water system, freeze-drying and lipid inversion method were successively used to produce the ARM-NPs for pMDI. Various characteristics of ARM-NPs were investigated including particle size, morphology, secondary structure of the peptide drug, aerosolization properties and storage stability. As revealed by the results, ARM-NPs with spherical shape possessed 147.7 ± 2.0 nm of particle size with 0.152 ± 0.021 PdI. The ARM-NPs for pMDI had satisfactory fine particle fraction (FPF) value of 46.99 ± 1.33%, while the secondary structure of the peptide drug was unchanged. Stability tests showed no pronounced sedimentation instability for over 12 weeks at 4-6 °C. Furthermore, a hypothesis was raised to explain the formation mechanism of ARM-NPs, which was verified by the differential scanning calorimetry analysis. The lecithin employed in the reverse micelle vesicles could serve as a steric barrier between peptide drugs and bulk propellant, which prevented the instability of peptide drugs in hydrophobic environment. Homogenous particle size could avoid Ostwald ripening phenomenon of particles in pMDIs. It was concluded that the ARM-NPs for pMDI could successfully overcome sedimentation instability by the steric barrier effect and homogeneous particle size.

[In vitro study of the effect of 11 kinds of natural drugs on the growth and acid production of Lactobacillus].

PURPOSE: To study the effect of 11 different natural drugs on the growth and acid production of Lactobacillus, as a preparation for screening an effective agent to mediate the balance of oral microflora. METHODS: Lactobacillus AC413 was chosen as the experimental bacterium. Eleven kinds of drugs, such as Rhizoma Chuanxiong, Surgentodoxa cuneata and Galla Chinensis were extracted by means of maceration, percolation and reflux extraction. The values of MIC of various extracts were measured. Then, different experimental media containing various extracts were prepared. The concentration of the extracts was lower than the MIC of the drug and the initial pH of the medium was 7.4. Lactobacillus was cultured in the medium for 48 hours, and finally the rest pH was measured. One-way ANOVA was used for statistical analysis. RESULTS: When the concentration of the drugs was lower than 8.000mg/ml, Tea polyphenols, Catechu, Galla Chinensis, Radix et Rhizoma Rhei and Nidus Vespae can inhibit the growth of Lactobacillus effectively. Tea polyphenols, Nidus Vespae, Radix Scuteilariae, Galla Chinensis and Surgentodoxa cuneata can inhibit the acid production of Lactobacillus effectively, Rhizoma Chuanxiong, Radix et Rhizoma Rhei, Semen Arecae and Catechu have no preliminary effect on it, but Surgentodoxa Cuneata and Radix Angelicae Pubescentis can increase it. CONCLUSION: Tea polyphenols, Catechu, Galla Chinensis, Radix et Rhizoma Rhei and Nidus Vespae can inhibit the growth of Lactobacillus effectively, and Tea polyphenols, Radix et Rhizoma Rhei, Nidus Vespae, Radix Scuteilariae, Galla Chinensis and Surgentodoxa Cuneata can inhibit the acid production of Lactobacillus effectively.

[The effects of natural medicine on adherence of Streptococcus mutans to salivary acquired pellicle].

OBJECTIVE: To investigate the effects of 5 kinds of natural medicine such as Radix et Rhizoma Rhei on the adherence of Streptococcus mutans to salivary acquired pellicle, and to screen effective natural medicines for the prevention of caries. METHODS: The in vitro model of experimental pellicle was saliva-coated hydroxyapatites (S-HA). Streptococcus mutans and S-HA were treated with natural medicines respectively, and then the adherence of bacteria to S-HA was tested. RESULTS: Radix et Rhizoma Rhei, Semen Arecae and Rhizoma Liguistici Chuanxiong could decrease the attachment of Streptococcus mutans to S-HA, and Semen Arecae was the most effective one. Radix Angelicae Dahuricae had no remarkable effect, and Catechu inhibited the adherence only after the bacteria being under treatment. CONCLUSION: Radix et Rhizoma Rhei, Semen Arecae, Rhizoma Liquistici Chuanxiong and Catechu could effectively inhibit the adherence of Streptococcus mutans to salivary acquired pellicle, but the effect of Radix Angelicae Dahuricae on the adherence is un-noticeable.

[Evaluation of in vitro cario-static effect of Galla chinensis with biofilm model].

OBJECTIVE: To evaluate the in vitro cario-static effect of Galla chinesis with biofilm model. METHODS: A four-organism bacterial consortium was cultured in a biofilm model on hydroxyapatite (HA) discs in a continuous culture system and exposed to repeated solution pulsing. There were three groups with different solution pulsed in the model: negative control group was pulsed with distilled water, positive control group was pulsed with 100 mmol/L sucrose solution and experimental group was pulsed with 100 mmol/L sucrose solution containing 4.0 g/L Galla chinensis. During the experiment, the dynamic changes of pH were recorded. After 6 pulses, surface structure of the biofilm was observed with a scanning electron microscope and the population on the biofilm was enumerated. RESULTS: Galla chinesis significantly inhibited the adherence of Actinomyces naelundii to HA disc compared with the control group and facilitated the removal of acid products. It was also found that the extra-cellular polysaccharide was reduced with the pulsing of Galla chinesis. CONCLUSION: Galla chinesis in the biofilm model can partially reduce the cario-genic response of sucrose solution.

[The effects of traditional Chinese medicines on the adherence of Streptococcus mutans to salivary acquired pellicle in vitro].

OBJECTIVES: To investigate the effects of traditional Chinese medicines on the preliminary adherence of Streptococcus mutans to salivary acquired pellicles. METHODS: The hydroxyapatites beads were coated by saliva to form S-HA, an in vitro model of experimental pellicle. Streptococcus mutans was chosen as the experiment strain, and radiolabeled with 3H-thymidine. Two parts of adherence inhibit study were carried on with medicines pretreated S-HA and medicines pretreated bacteria. The numbers of 3H-thymidine labeled bacteria attached to S-HA were counted by scintillation counting to indicate the amount of adherence. RESULTS: It was found that most of the traditional Chinese medicines tested could inhibit the adherence to S-HA to different extent, especially the Galla Chinensis, which has significant inhibitory action. CONCLUSION: Galla Chinensis may be a prospective medicine to inhibit the cariogenic bacteria's preliminary adherence to salivary acquired pellicles.

[The in vitro study of the effects of 11 kinds of traditional Chinese medicine on the growth and acid production of Actinomyces viscosus].

OBJECTIVE: To assess the effects of different natural medicines on the growth and acid production of Actinomyces viscosus, thus making preparations for screening an effective agent to mediate the balance of oral microflora. METHODS: Actinomyces viscosus ATCC 19246 was chosen as the experimental bacteria. 11 kinds of traditional Chinese medicine, such as Rhizoma Ligustici Chuanxiong, Sargentodoxa Cuneata and Galla Chinensis were extracted by means of maceration, percolation and reflux extraction. First, the values of MIC of various extracts were measured. Second, the experimental medium containing various extracts was prepared. The concentration of the extracts was lower than the MIC of the medicine, and the initial pH of the medium was 7.4. Then Actinomyces viscosus was cultured in the medium for 48 h, and finally the rest pH was measured. RESULTS: When the concentration of the medicines was lower than or equal to 8.000 mg/ml, it was found that all kinds of medicine except Radix Notoginseng can inhibit the growth of Actinomyces viscosus effectively, especially Polistes mandarinus and Semen Arecae. Tea polyphenols, Radix Notoginseng, Radix et Rhizoma Rhei, Polistes mandarinus and Sargentodoxa cuneata can inhibit the acid production of Actinomyces viscosus effectively, but Radix Scutellariae, Rhizoma Ligustici Chuanxiong, Semen Arecae, Radix Angelicae Dahuricae, Galla Chinensis and Catechu have no preliminary effect on it. CONCLUSION: Tea polyphenols, Radix et Rhizoma Rhei, Polistes mandarinus and Sargentodoxa cuneata can inhibit the growth and the acid production of Actinomyces viscosus effectively.