Discovery of Gambogic acid as an antibacterial adjuvant against vancomycin-resistant enterococci in vitro and in vivo.

BACKGROUND: The emergence and spread of vancomycin-resistant enterococci (VRE) have posed a significant challenge to clinical treatment, underscoring the need to develop novel strategies. As therapeutic options for VRE are limited, discovering vancomycin enhancer is a feasible way of combating VRE. Gambogic acid (GA) is a natural product derived from the resin of Garcinia hanburyi Hook.f. (Clusiaceae), which possesses antibacterial activity. PURPOSE: This study aimed to investigate the potential of GA as an adjuvant to restore the susceptibility of VRE to vancomycin. METHODS: In vitro antibacterial and synergistic activities were evaluated against vancomycin-susceptible and resistant strains by the broth microdilution method for the Minimal Inhibitory Concentrations (MICs) determination, and checkerboard assay and time-kill curve analysis for synergy evaluation. In vivo study was conducted on a mouse multi-organ infection model. The underlying antibacterial mechanism of GA was also explored. RESULTS: GA showed a potent in vitro activity against all tested strains, with MICs ranging from 2 to 4 µg/ml. The combination of GA and vancomycin exhibited a synergistic effect against 18 out of 23 tested VRE strains, with a median fractional inhibitory concentration index (FICI) of 0.254, and demonstrated a synergistic effect in the time-kill assay. The combination therapy exhibited a significant reduction in tissue bacterial load compared with either compound used alone. GA strongly binds to the ParE subunit of topoisomerase IV, a bacterial type II DNA topoisomerase, and suppresses its activity. CONCLUSIONS: The study suggests that GA has a significant antibacterial activity against enterococci, and sub-MIC concentrations of GA can restore the activity of vancomycin against VRE in vitro and in vivo. These findings indicate that GA has the potential to be a new antibacterial adjuvant to vancomycin in the treatment of infections caused by VRE.

Acupuncture and Doxylamine-Pyridoxine for Nausea and Vomiting in Pregnancy : A Randomized, Controlled, 2 × 2 Factorial Trial.

BACKGROUND: An effective and safe treatment for nausea and vomiting of pregnancy (NVP) is lacking. OBJECTIVE: To assess the efficacy and safety of acupuncture, doxylamine-pyridoxine, and a combination of both in women with moderate to severe NVP. DESIGN: Multicenter, randomized, double-blind, placebo-controlled, 2 × 2 factorial trial. (ClinicalTrials.gov: NCT04401384). SETTING: 13 tertiary hospitals in mainland China from 21 June 2020 to 2 February 2022. PARTICIPANTS: 352 women in early pregnancy with moderate to severe NVP. INTERVENTION: Participants received daily active or sham acupuncture for 30 minutes and doxylamine-pyridoxine or placebo for 14 days. MEASUREMENTS: The primary outcome was the reduction in Pregnancy-Unique Quantification of Emesis (PUQE) score at the end of the intervention at day 15 relative to baseline. Secondary outcomes included quality of life, adverse events, and maternal and perinatal complications. RESULTS: No significant interaction was detected between the interventions (P = 0.69). Participants receiving acupuncture (mean difference [MD], -0.7 [95% CI, -1.3 to -0.1]), doxylamine-pyridoxine (MD, -1.0 [CI, -1.6 to -0.4]), and the combination of both (MD, -1.6 [CI, -2.2 to -0.9]) had a larger reduction in PUQE score over the treatment course than their respective control groups (sham acupuncture, placebo, and sham acupuncture plus placebo). Compared with placebo, a higher risk for births with children who were small for gestational age was observed with doxylamine-pyridoxine (odds ratio, 3.8 [CI, 1.0 to 14.1]). LIMITATION: The placebo effects of the interventions and natural regression of the disease were not evaluated. CONCLUSION: Both acupuncture and doxylamine-pyridoxine alone are efficacious for moderate and severe NVP. However, the clinical importance of this effect is uncertain because of its modest magnitude. The combination of acupuncture and doxylamine-pyridoxine may yield a potentially larger benefit than each treatment alone. PRIMARY FUNDING SOURCE: The National Key R&D Program of China and the Project of Heilongjiang Province "TouYan" Innovation Team.

In Situ Synthesized Homochiral Spiroborate Ester Metal-Organic Framework with Mono-, Di-, and Trivalent Cations.

We report here a one-step method for synthesizing multi-component and in-situ-formed homochiral spiroborate-ester-based metal-organic framework CPM-B1. This unique material successfully integrates COF fragment spiroborate ester within the MOF and simultaneously incorporate homochirality and helicity. In addition, CPM-B1 is a rare example of framework materials that results from the cooperative assembly of three charge-complementary cations: +1 (lithium), +2 (cobalt), and +3 (boron). The sophistication of the co-assembly is further highlighted by the three structural roles of lithium ions. This unique structure contributes to its multi-functional properties such as ionic conductivity and catalytic activity for oxygen reduction reaction (by CPM-B1 carbonized material) and provides a new path to develop MOF materials with complex secondary building units and multi-functional applications.

Berberine Directly Targets the NEK7 Protein to Block the NEK7-NLRP3 Interaction and Exert Anti-inflammatory Activity.

Berberine (BBR), a traditional Chinese medicine, has therapeutic effects on a variety of inflammation-related diseases, but its direct proteomic targets remain unknown. Using activity-based protein profiling, we first demonstrated that BBR directly targets the NEK7 protein via the hydrogen bond between the 2,3-methylenedioxy and 121-arginine (R121) residues. The fact that R121 is located precisely within the key domain involved in the NEK7-NLRP3 interaction allows BBR to specifically block the NEK7-NLRP3 interaction and successively inhibit IL-1ß release, independent of the NF-κB and TLR4 signaling pathways. Moreover, BBR displays in vivo anti-inflammatory efficacy in a NEK7-dependent manner. Therefore, we consider NEK7 to be a key target of BBR in the treatment of NLRP3-related inflammatory diseases, and the development of novel NEK7-NLRP3 interaction inhibitors might be easily achieved using NEK7 as a target.

[Effect of ligustilide on oxygen and glucose deprivation/reperfusion-induced mitochondria fission in PC12 cells].

This study aimed to investigate the effect and mechanism of ligustilide, the main active ingredient in Ligusticum wallichii, on mitochondria fission after PC12 cell injury induced by oxygen and glucose deprivation/reperfusion(OGD/R). In the experiment, an OGD/R model was established in vitro, and PC12 cells were pre-treated with ligustilide for 3 h, and then the cell viability was detected by CCK-8 method. The effect of different concentrations of ligustilide on the morphology of PC12 cells after OGD/R injury was observed under an inverted microscope. Transmission electron microscopy was used to observe the mitochondrial fission of PC12 cells after OGD/R injury. DCFH-DA immunofluorescence staining method was used to detect intracellular reactive oxygen species(ROS) changes. Changes in mitochondria membrane potential(MMP) were detected by flow cytometry. Hochest 33258 was used to observe the apoptosis of PC12 cells. Western blot was used to detect changes in cytochrome C(Cyt C) content in mitochondria and cytoplasm, and mitochondrial fission-related proteins Drp 1 and Fis 1. All results showed that compared with the model group, ligustilide significantly increased the survival rate of PC12 cells and the number of cells. Further experiments showed that ligustilide inhibited the release of ROS and decline of mitochondrial membrane potential in PC12 cells after OGD/R injury. Moreover, ligustilide reduced the release of Cyt C and promoted the expressions of Drp1 and Fis1 in mitochondrial fission proteins. Verification experiments showed that mitochondrial fission inhibitor mdivi-1 decreased cell survival rate and inhibited fission. The results indicated that ligustilide exerted neuro-protective effects by promoting mitochondrial fission and reducing cell damage. It preliminary proves that the mechanism of ligustilide on ischemic brain injury may be related to the promotion of mitochondrial fission and the maintenance of cell homeostasis.

Lock-and-Key and Shape-Memory Effects in an Unconventional Synthetic Path to Magnesium Metal-Organic Frameworks.

We report a new magnesium metal-organic framework (MOF) (CPM-107) with a special interaction with CO2 . CPM-107 contains Mg2 -acetate chains crosslinked into a 3D net by terephthalate. It has an anionic framework encapsulating ordered extra-framework cations and solvent molecules. The desolvated form is closed and unresponsive to common gasses, such as N2 , H2 , and CH4 . Yet, with CO2 at 195 K, it abruptly opens and turns into a rigid porous form that is irreversible via desorption. Once opened by CO2 , CPM-107 remains in the stable porous state accessible to additional gas types over multiple cycles or CO2 itself at different temperatures. The porous phase can be re-locked to return to the initial closed phase via re-solvation and desolvation. Such peculiar properties of CPM-107 are apparently linked to a convergence of factors related to both framework and extra-framework features. The unusual CO2 effect is currently the only available path to porous CPM-107 which shows efficient C2 H2 /CO2 separation.

Evolution of matrinic ethanol derivatives as anti-HCV agents from matrine skeleton.

Twenty-two novel 12N-substituted matrinic ethanol derivatives were synthesized and evaluated for their antiviral activities against HCV taking compound 1 as the lead. The SAR study indicated that the shortening of the 11-butyl chain to ethyl chain did not affect the activity significantly. Out of the target compounds, matrinic ethanol 6a demonstrated a potential anti-HCV effect with an EC50 value of 3.2µM and a SI value of 96.6. The free hydroxyl arm in 6a made it possible as a parent structure to prepare pro-drug for the potential application in HCV treatment. This study provided powerful information on further strategic optimization and development of this kind of compounds into a novel family of anti-HCV agents.

Tetrandrine potentiates the hypoglycemic efficacy of berberine by inhibiting P-glycoprotein function.

This study was designed to improve the absorption and hypoglycemic efficacy of berberine (BBR), which is a substrate of P-glycoprotein (P-gp), by combination with a P-gp inhibitor tetrandrine (Tet). Flow cytometry and LC-MS/MS were used to determine the cellular efflux or retention of chemicals. Pharmacokinetic study was performed in ICR mice following oral administration of the study compounds. The hypoglycemic efficacies of the compounds were evaluated in diabetic KK-Ay mice. In the in vitro experiments, Tet significantly inhibited the efflux and increased the uptake of P-gp substrates rhodamine-123 as well as BBR in MCF7/DOX cells and Caco-2 intestinal cells. Meanwhile, Tet greatly reduced the expression of P-gp in Caco-2 cells. The inhibition of BBR efflux by Tet was translated into improved pharmacokinetics in vivo. When co-administered, Tet dose-dependently increased the average maximum concentration (C(max)) and area under concentration-time curve (AUC0₋24) of BBR in mice. Tet itself had no impact on glucose metabolism. However, it greatly potentiated the hypoglycemic efficacy of BBR in diabetic KK-Ay mice. In addition, we found that Tet had moderate inhibitory effect on the catalytic activity of CYP3A4, which played a role in the bio-transformation of BBR, and this may also take part in the improvement of the pharmacokinetics of BBR. In summary, combination with P-gp inhibitors such as Tet can improve the pharmacokinetics and hypoglycemic efficacy of BBR greatly; this implicates a feasible strategy for exploring the therapeutic effects of BBR and other pharmaceuticals which are substrates of P-gp.

[Comparison of therapeutic effects between plum-blossom needle tapping plus cupping and laser irradiation in the treatment of acute facial palsy patients with concomitant peri-auricular pain].

OBJECTIVE: To compare the therapeutic effects of plum-blossom needle therapy with cupping and laser irradiation in the treatment of acute facial paralysis with ipsilateral peri-auricular pain. METHODS: Sixty outpatients with acute facial paralysis and ipsilateral peri-auricular pain were divided into plum-blossom needle (treatment) group (n = 28) and laser-irradiation (control) group (n = 32). In the acute stage, patients of the treatment group were treated with plum-blossom needle tapping in combination with cupping at Yifeng (TE 17) and Wangu (GB 12), while those of the control group treated with He-Ne laser irradiation over the same two acupoints until the peri-auricular pain disappeared. The treatment was given once daily. Following disappearance of the pain, routine acupuncture treatment of Yifeng (TE 17), Wangu (GB 12), Dicang (ST 4), etc. was given continuously to patients of the two groups, once daily for 20 times. The treatment times for healing ipsilateral peri-auricular pain, and the electromyogram (EMG, compound muscle action potential) of the ipsilateral musculus orbicularis oris were recorded. Scores of the facial nerve function were given to House-Brackmann Facial Nerve Grading System. RESULTS: The treatment times for healing peri-auricular pain in the treatment group and laser irradiation group were 2.9 +/- 1. 0 and 6.0 +/- 2.2 respectively (t = 6.816, P = 0.000). The scores of House-Brackmann scale of the paralyzed side in the treatment group and control group were (3.3 +/- 1.5) points and (3.3 +/- 1.4) points (P > 0.05) before the treatment; (1.8 +/- 1.1) points and (2.5 +/- 1.2) points (P < 0.05) one month after the treatment, and (1.5 +/- 0.9) points and (2.2 +/- 1.5) points (P < 0.05) three months after the treatment, respectively. The amplitude ratios of compound muscle action potential between the affected side and the healthy side in the treatment group and control group were (52.1 +/- 20.5)% and (51.2 +/- 22.6)% (P > 0.05) during the first two weeks after onset, and (79.2 +/- 11.3)% and (69.8 +/- 17.9)% (P < 0.05) three months after the treatment, respectively. CONCLUSION: Plum-blossom needle tapping plus cupping is significantly superior to that of He-Ne laser irradiation in reducing the treatment sessions for relieving peri-auricular pain during acute stage, and improving facial and muscular functions in the treatment of acute facial paralysis patients.

[Observation on therapeutic effects of acupuncture combined with TDP irradiation and chinese herbal steaming and washing therapy for treatment of carpal tunnel syndrome in early stage].

OBJECTIVE: To search for an effective therapy for carpal tunnel syndrome in early stage. METHODS: Sixty cases were randomly divided into an observation group 1 (21 cases), an observation group 2 (22 cases) and a basic treatment group (17 cases). The patients in three groups were treated with oral administration of Vitamin B1, Vitamin B12, compound Vitamin B and small splint for wrist protection as basic treatment, and the observation group 1 was also treated with acupuncture on local points as Laogong (PC 8), Yuji (LU 10), Hegu (LI 4) and Waiguan (TE 5) etc. combined with TDP irradiation, and the observation group 2 was treated with herbal steaming and washing on the affected part using Safflower and Lopseed etc. Electromyography, visual analogue scale (VAS) and clinical effect of three groups were observed before and after treatment. RESULTS: The changes of electromyogra phy had no significant difference before and after treatment in all groups (all P > 0.05), the VAS scores in two ob servation groups were superior to that in basic treatment group (both P < 0.05). The excellent and good rates were 90.5% and 86.4% in the observation group 1 and the observation group 2 respectively, both higher than that of 64.7% in the basic treatment group. CONCLUSION: Acupunture combined with TDP irradiation and Chinese herbal steaming and washing therapy both have significant therapeutic effects for treatment of carpal tunnel syndrome in early stage.

[Epidemic survey on TCM syndrome typing and preliminary discussion on TCM etiology and pathogenesis of fatty liver].

OBJECTIVE: To investigate the common TCM syndrome types of fatty liver by way of epidemic questionnaire, their occurrence ratio, and the correlation between various syndrome types and objective indexes. METHODS: A total of national wide 503 subjects with fatty liver were enrolled, the TCM syndromes, body mass index (BMI), abdominal perimeter/hip circumference, liver function, blood lipids, B ultrasonic examination and CT in them were observed and recorded. RESULTS: In the 46 symptoms investigated in total, the first ten symptoms in order of appearing rate were lassitude, obese, oral dryness, vertigo, hypochondriac distending pain, soreness and pain in loin, spiritlessness, oral bitterness, aching and weakness in knee and abdominal distention. The mostly appeared tongue figures were pale and corpulent or pale dim tongue proper, white greasy or yellow greasy tongue coating, and the mostly appeared pulse figures were taut, taut-thin and taut slippery. Statistical cluster analysis showed that syndromes of fatty liver could be typed into 4 TCM types, the asthenia Pi-Shen with Gan-stagnation type, the asthenia Pi-Shen type, the asthenia Pi with phlegm-heat type and the unclassified type. Among them the asthenia Pi-Shen with Gan-stagnation type was the commonest one, which accounted to 62.32%. CONCLUSION: The mostly appeared syndrome type of fatty liver was asthenia Pi-Shen with Gan-stagnation type. The TCM pathogenesis of fatty liver was deficiency of origin, mainly deficiency of Shen, involving Pi, with excess superficiality, the turbid-phlegm and blood stasis.