The metabolic slowdown caused by the deletion of pspA accelerates protein aggregation during stationary phase facilitating antibiotic persistence.

Entering a dormant state is a prevailing mechanism used by bacterial cells to transiently evade antibiotic attacks and become persisters. The dynamic progression of bacterial dormancy depths driven by protein aggregation has been found to be critical for antibiotic persistence in recent years. However, our current understanding of the endogenous genes that affects dormancy depth remains limited. Here, we discovered a novel role of phage shock protein A (pspA) gene in modulating bacterial dormancy depth. Deletion of pspA of Escherichia coli resulted in increased bacterial dormancy depths and prolonged lag times for resuscitation during the stationary phase. ∆pspA exhibited a higher persister ratio compared to the wild type when challenged with various antibiotics. Microscopic images revealed that ∆pspA showed accelerated formation of protein aggresomes, which were collections of endogenous protein aggregates. Time-lapse imaging established the positive correlation between protein aggregation and antibiotic persistence of ∆pspA at the single-cell level. To investigate the molecular mechanism underlying accelerated protein aggregation, we performed transcriptome profiling and found the increased abundance of chaperons and a general metabolic slowdown in the absence of pspA. Consistent with the transcriptomic results, the ∆pspA strain showed a decreased cellular ATP level, which could be rescued by glucose supplementation. Then, we verified that replenishment of cellular ATP levels by adding glucose could inhibit protein aggregation and reduce persister formation in ∆pspA. This study highlights the novel role of pspA in maintaining proteostasis, regulating dormancy depth, and affecting antibiotic persistence during stationary phase.

Spinal cord injury-induced gut dysbiosis influences neurological recovery partly through short-chain fatty acids.

Spinal cord injury (SCI) can reshape gut microbial composition, significantly affecting clinical outcomes in SCI patients. However, mechanisms regarding gut-brain interactions and their clinical implications have not been elucidated. We hypothesized that short-chain fatty acids (SCFAs), intestinal microbial bioactive metabolites, may significantly affect the gut-brain axis and enhance functional recovery in a mouse model of SCI. We enrolled 59 SCI patients and 27 healthy control subjects and collected samples. Thereafter, gut microbiota and SCFAs were analyzed using 16 S rDNA sequencing and gas chromatography-mass spectrometry, respectively. We observed an increase in Actinobacteriota abundance and a decrease in Firmicutes abundance. Particularly, the SCFA-producing genera, such as Faecalibacterium, Megamonas, and Agathobacter were significantly downregulated among SCI patients compared to healthy controls. Moreover, SCI induced downregulation of acetic acid (AA), propionic acid (PA), and butyric acid (BA) in the SCI group. Fecal SCFA contents were altered in SCI patients with different injury course and injury segments. Main SCFAs (AA, BA, and PA) were administered in combination to treat SCI mice. SCFA supplementation significantly improved locomotor recovery in SCI mice, enhanced neuronal survival, promoted axonal formation, reduced astrogliosis, and suppressed microglial activation. Furthermore, SCFA supplementation downregulated NF-κB signaling while upregulating neurotrophin-3 expression following SCI. Microbial sequencing and metabolomics analysis showed that SCI patients exhibited a lower level of certain SCFAs and related bacterial strains than healthy controls. SCFA supplementation can reduce inflammation and enhance nourishing elements, facilitating the restoration of neurological tissues and the improvement of functional recuperation. Trial registration: This study was registered in the China Clinical Trial Registry ( www.chictr.org.cn ) on February 13, 2017 (ChiCTR-RPC-17010621).

Total Glucosides of Paeony Regulate Immune Imbalance Mediated by Dermal Mesenchymal Stem Cells in Psoriasis Mice.

OBJECTIVE: To investigate the therapeutic effects of total glucosides of paeony (TGP) on psoriasis based on the immunomodulatory effect of dermal mesenchymal stem cells (DMSCs). METHODS: A total of 30 male BALB/c mice were divided into 6 groups (n=5 in each) by a random number table method, including control, psoriasis model (model, 5% imiquimod cream 42 mg/d), low-, medium- and high-dose TGP (50, 100, and 200 mg/kg, L, M-, and H-TGP, respectively), and positive control group (2.5 mg/kg acitretin). After 14 days of continuous administration, the skin's histopathological changes, apoptosis, secretion of inflammatory cytokines, and proportion of regulatory T cells (Treg) and T helper cell 17 (Th17) were evaluated using hematoxylin-eosin (HE) staining, TdT-mediated dUTP nick end labeling staining, enzyme-linked immunosorbent assay, and flow cytometry, respectively. DMSCs were further isolated from the skin tissues of normal and psoriatic mice, and the cell morphology, phenotype, and cycle were observed. Furthermore, TGP was used to treat psoriatic DMSCs to analyze the effects on the DMSCs immune regulation. RESULTS: TGP alleviated skin pathological injury, reduced epidermis layer thickness, inhibited apoptosis, and regulated the secretion of inflammatory cytokines and the proportion of Treg and Th17 in the skin tissues of psoriatic mice (P<0.05 or P<0.01). There was no significant difference in cell morphology and phenotype between control and psoriatic DMSCs (P>0.05), however, more psoriatic DMSCs remained in G0/G1 phase compared with the normal DMSCs (P<0.01). TGP treatment of psoriatic DMSCs significantly increased cell viability, decreased apoptosis, relieved inflammatory response, and inhibited the expression of toll-like receptor 4 and P65 (P<0.05 or P<0.01). CONCLUSION: TGP may exert a good therapeutic effect on psoriasis by regulating the immune imbalance of DMSCs.

Enhanced Effect of Qing-Ying Decoction Concurrent with Secukinumab in the Treatment of Plaque Psoriasis.

Both Qing-Ying decoction (QYD), a Traditional Chinese Medicine (TCM), and secukinumab, a fully humanized anti-interleukin-17A monoclonal antibody, have been used to treat patients with plaque psoriasis. The combined application of TCM and biologics in the treatment of psoriasis, however, has not been investigated. We enrolled a total of 68 patients with plaque psoriasis in our prospective study, and randomly assigned them to either the study group (treated with secukinumab plus QYD), or the control group (treated with secukinumab alone). After 12- and 16-week treatment, the Psoriasis Area and Severity Index (PASI) score and the TCM score were significantly reduced in both the study and the control groups. However, the reduction in PASI and TCM scores was more significant in the study group than in the control group (12-week: PASI: 5.29 ± 0.27 vs 8.87 ± 0.38, respectively; P < .01; TCM: 5.83 ± 0.21 vs 12.39 ± 1.23, respectively; P < .01; 16-week: PASI score: 4.76 ± 0.18 vs 8.36 ± 0.31, respectively; TCM score: 4.98 ± 0.19 vs 11.27 ± 1.13, respectively; P < .01). The total treatment efficacy rate was significantly higher in the study group (97.1%) than the control group (76.5%; P = .012). The number of CD3+ and CD4+ T cells was increased, while the number of CD8+ T cells was decreased after treatment in both groups, with more significant changes in the study group (P < .01). QYD may enhance the therapeutic outcome of secukinumab in the treatment of plaque psoriasis by further suppressing chronic skin inflammation, as well as reducing adverse events and patients' psychological stress.

Fecal Microbiota Transplantation Exerts Neuroprotective Effects in a Mouse Spinal Cord Injury Model by Modulating the Microenvironment at the Lesion Site.

The primary traumatic event that causes spinal cord injury (SCI) is followed by a progressive secondary injury featured by vascular disruption and ischemia, inflammatory responses and the release of cytotoxic debris, which collectively add to the hostile microenvironment of the lesioned cord and inhibit tissue regeneration and functional recovery. In a previous study, we reported that fecal microbiota transplantation (FMT) promotes functional recovery in a contusion SCI mouse model; yet whether and how FMT treatment may impact the microenvironment at the injury site are not well known. In the current study, we examined individual niche components and investigated the effects of FMT on microcirculation, inflammation and trophic factor secretion in the spinal cord of SCI mice. FMT treatment significantly improved spinal cord tissue sparing, vascular perfusion and pericyte coverage and blood-spinal cord-barrier (BSCB) integrity, suppressed the activation of microglia and astrocytes, and enhanced the secretion of neurotrophic factors. Suppression of inflammation and upregulation of trophic factors, jointly, may rebalance the niche homeostasis at the injury site and render it favorable for reparative and regenerative processes, eventually leading to functional recovery. Furthermore, microbiota metabolic profiling revealed that amino acids including ß-alanine constituted a major part of the differentially detected metabolites between the groups. Supplementation of ß-alanine in SCI mice reduced BSCB permeability and increased the number of surviving neurons, suggesting that ß-alanine may be one of the mediators of FMT that participates in the modulation and rebalancing of the microenvironment at the injured spinal cord. IMPORTANCE FMT treatment shows a profound impact on the microenvironment that involves microcirculation, blood-spinal cord-barrier, activation of immune cells, and secretion of neurotrophic factors. Analysis of metabolic profiles reveals around 22 differentially detected metabolites between the groups, and ß-alanine was further chosen for functional validation experiments. Supplementation of SCI mice with ß-alanine significantly improves neuronal survival, and the integrity of blood-spinal cord-barrier at the lesion site, suggesting that ß-alanine might be one of the mediators following FMT that has contributed to the recovery.

Single Cell RNA Sequencing Identifies a Unique Inflammatory Macrophage Subset as a Druggable Target for Alleviating Acute Kidney Injury.

Acute kidney injury (AKI) is a complex clinical disorder associated with poor outcomes. Targeted regulation of the degree of inflammation has been a potential strategy for AKI management. Macrophages are the main effector cells of kidney inflammation. However, macrophage heterogeneity in ischemia reperfusion injury induced AKI (IRI-AKI) remains unclear. Using single-cell RNA sequencing of the mononuclear phagocytic system in the murine IRI model, the authors demonstrate the complementary roles of kidney resident macrophages (KRMs) and monocyte-derived infiltrated macrophages (IMs) in modulating tissue inflammation and promoting tissue repair. A unique population of S100a9hi Ly6chi IMs is identified as an early responder to AKI, mediating the initiation and amplification of kidney inflammation. Kidney infiltration of S100A8/A9+ macrophages and the relevance of renal S100A8/A9 to tissue injury is confirmed in human AKI. Targeting the S100a8/a9 signaling with small-molecule inhibitors exhibits renal protective effects represented by improved renal function and reduced mortality in bilateral IRI model, and decreased inflammatory response, ameliorated kidney injury, and improved long-term outcome with decreased renal fibrosis in the unilateral IRI model. The findings support S100A8/A9 blockade as a feasible and clinically relevant therapy potentially waiting for translation in human AKI.

A Meta-Analysis of the Association between Diabetes Mellitus and Traditional Chinese Medicine Constitution.

OBJECTIVE: To explore the distribution of constitution types of diabetes mellitus (DM) in traditional Chinese medicine (TCM) and to provide evidence-based medicine basis for the prevention and treatment of diabetes. METHODS: PubMed, Embase, Web of Science, and three Chinese databases were searched to include research literature on the relationship between diabetes and TCM constitution. The single rate study of cross-sectional literature was conducted with RStudio software, and the control meta-analysis of the diabetic and nondiabetic population was performed with Review Manager 5.3 software. Two independent reviewers assessed the methodological quality of the studies' data. The main outcomes included the distribution of constitutional types in the diabetic population and the odds ratio (OR) between the two. Effect sizes are expressed as proportions or ORs with 95% confidence intervals (CI). RESULTS: A total of 28,781 diabetic cases were included in 87 articles. Yin-deficiency, phlegm-dampness, and qi-deficiency accounted for 18% (95% CI (15%, 20%), P < 0.01), 17% (95% CI (15%, 19%), P < 0.01), and 13% (95% CI (11%, 15%), P < 0.01) of the total diabetic cases. The risk of diabetes in people with yin-deficiency and phlegm-dampness was 3.06 (95% CI (1.38-6.78), P=0.006) and 1.89 (95%CI (1.05-3.42), P=0.03) times higher than that in those with other constitutions, respectively. The distribution of TCM constitution of DM patients varied significantly in different regions and ages. CONCLUSION: Yin-deficiency and phlegm-dampness are the common constitution types of diabetic people, and they may also be the risk factors of diabetes. Balanced constitution may be a protective factor of diabetes. More high-quality cohort and case-control studies need to be designed to provide more valuable evidence-based basis for assessing the correlation between DM and TCM constitution.

The oleaginous astaxanthin-producing alga Chromochloris zofingiensis: potential from production to an emerging model for studying lipid metabolism and carotenogenesis.

The algal lipids-based biodiesel, albeit having advantages over plant oils, still remains high in the production cost. Co-production of value-added products with lipids has the potential to add benefits and is thus believed to be a promising strategy to improve the production economics of algal biodiesel. Chromochloris zofingiensis, a unicellular green alga, has been considered as a promising feedstock for biodiesel production because of its robust growth and ability of accumulating high levels of triacylglycerol under multiple trophic conditions. This alga is also able to synthesize high-value keto-carotenoids and has been cited as a candidate producer of astaxanthin, the strongest antioxidant found in nature. The concurrent accumulation of triacylglycerol and astaxanthin enables C. zofingiensis an ideal cell factory for integrated production of the two compounds and has potential to improve algae-based production economics. Furthermore, with the advent of chromosome-level whole genome sequence and genetic tools, C. zofingiensis becomes an emerging model for studying lipid metabolism and carotenogenesis. In this review, we summarize recent progress on the production of triacylglycerol and astaxanthin by C. zofingiensis. We also update our understanding in the distinctive molecular mechanisms underlying lipid metabolism and carotenogenesis, with an emphasis on triacylglycerol and astaxanthin biosynthesis and crosstalk between the two pathways. Furthermore, strategies for trait improvements are discussed regarding triacylglycerol and astaxanthin synthesis in C. zofingiensis.

Paleolithic genetic link between Southern China and Mainland Southeast Asia revealed by ancient mitochondrial genomes.

The genetic history of Southern East Asians is not well-known, especially prior to the Neolithic period. To address this, we successfully sequenced two complete mitochondrial genomes of 11,000-year-old human individuals from Southern China, thus generating the oldest ancient DNA sequences from this area. Integrating published mitochondrial genomes, we characterized M71d, a new subhaplogroup of haplogroup M71. Our results suggest a possible early migration between Southern China and mainland Southeast Asia by at least 22,000 BP.

Balneotherapy with Chinese herbal medicine prolongs the remission period in patients with psoriasis vulgaris.

This study aimed to evaluate whether the supplementary balneotherapy with Chinese herbal medicine (CHM) could facilitate the treatment of psoriasis vulgaris and thus be beneficial for long-term remission from the symptoms. Two hundred psoriasis vulgaris patients with moderate-to-severe plaque psoriasis from January 2013 to June 2014 were evenly divided into two groups: the consolidated therapy group (CTG) and unconsolidated therapy group (UTG); the remission period of the two groups was compared. There was no significant difference in Psoriasis Area Severity Index (PASI) score between the two groups at the beginning and the end of the treatment. However, the average remission time in CTG was 10.99 months, which was significantly longer than that of 7.94 months in UTG (P = .001). After a correction of age, course of disease, skin type as well as PASI baseline value using a COX model, we found that the risk of recurrence of psoriasis vulgaris in UTG was higher than that in the CTG (P < .001). No adverse reactions were discovered when combing the two treatments together. The combined treatment of CHM balneotherapy and narrowband ultraviolet B could significantly prolong the remission time in patients with psoriasis vulgaris.

Anti-proliferative and apoptotic effects of S1, a tetrandrine derivative, in human gastric cancer BGC-823 cells.

The aim of the study was to investigate the anti-proliferation and apoptosis-inducing effects of S1, a novel tetrandrine derivative, in human gastric cancer BGC-823 cells and explore the possible mechanism of action. The anti-proliferative activity was determined by MTT assay; the induction of cell cycle arrest and apoptosis were detected by flow cytometry. Quantitative real time RT-PCR and Western blotting were used to evaluate the mRNA and protein expression levels in mitochondrial pathway. S1 significantly reduced cell viability and induced a G2/M phase arrest and apoptosis in dose- and time-dependent manner. Further studies showed that S1 increased mRNA and protein expression of Bax and the Bax/Bcl-2 ratio. Moreover, S1 decreased the protein expression of procaspase-9 and procaspase-3, suggesting that the induction of apoptosis may be related to the alteration of the ratio of Bax/Bcl-2 and the activation of caspases. These findings suggested that S1 merits further investigation as a novel therapeutic agent for the treatment of human gastric cancer.

Comparative pharmacokinetics and bioavailability of four alkaloids in different formulations from Corydalis decumbens.

ETHNOPHARMACOLOGICAL RELEVANCE: Corydalis decumbens, a Traditional Chinese Medicine listed in Chinese Pharmacopoeia, is clinically used for the treatment of paralytic stroke, headache, rheumatic arthritis and sciatica in China. AIM OF THE STUDY: This study was aimed to compare the pharmacokinetics and bioavailability of protopine, tetrahydropalmatine, bicuculline, and egenine in three formulations prepared from the rhizomes of Corydalis decumbens. MATERIALS AND METHODS: Alkaloid extract (CDAs-SFE) was prepared from the rhizomes of Corydalis decumbens by supercritical CO2 fluid extraction; CDAs-SFE/HPßCD (hydroxypropyl-ß-cyclodextrin inclusion complex), and CDAs-SFE/HCl (hydrochloride freeze-dried powder) were resulted from CDAs-SFE through complexation with HPßCD and hydrochloride, respectively. An UFLC-MS/MS method was developed for quantitative analysis of protopine, tetrahydropalmatine, bicuculline and egenine simultaneously in rat plasma after oral administration. The differences of pharmacokinetics and bioavailability of the four alkaloids in three formulations were determined by pharmacokinetics analyses. RESULTS AND CONCLUSIONS: The Cmax, AUC and bioavailability of protopine and tetrahydropalamatine (bioactive components) in CDAs-SFE/HCl were significantly higher than in CDAs-SFE and in CDAs-SFE/HPßCD. In contrast, in CDAs-SFE/HPßCD, AUC and bioavailability of tetrahydropalamatine were significantly lower, while those of bicuculline (toxic compound) appeared to be higher than both in CDAs-SFE and in CDAs-SFE/HCl. The results indicated that CDAs-SFE/HCl was the best beneficial formulation among the three formulations for the alkaloid extract prepared from the rhizomes of Corydalis decumbens, in which protopine and tetrahydropalamatine displayed higher bioavailability, but lower for bicuculline.

Calycosin and formononetin from astragalus root enhance dimethylarginine dimethylaminohydrolase 2 and nitric oxide synthase expressions in Madin Darby Canine Kidney II cells.

Nitric oxide (NO) is a crucial vasodilator produced by nitric oxide synthase (NOS). Asymmetric dimethylarginine (ADMA) is an endogenous NOS inhibitor and mainly catabolized by dimethylarginine dimethylaminohydrolase (DDAH). As we reported, the antihypertensive effect of shichimotsukokato (SKT), a formula of Japanese traditional kampo medicine consisting of 7 crude drugs, in 5/6 nephrectomized rats, is mediated by the DDAH-ADMA-NO pathway. Our present study aimed to explore the effective compounds of SKT using Madin Darby Canine Kidney (MDCK) II cells. We isolated two isoflavones, calycosin and formononetin from astragalus root, one of the components of SKT, which can promote DDAH2 protein and mRNA expressions in MDCK II cells. The neuronal NOS levels were also upregulated by the treatment of calycosin and formononetin. These results suggest that calycosin and formononetin could be the active ingredients of astragalus root and SKT that cause antihypertensive effects. The increased levels of DDAH2 and NOS may enhance NO production, decrease ADMA level and improve endothelial and cardiovascular dysfunction.

Anti-hypertensive effects of shichimotsukokato in 5/6 nephrectomized Wistar rats mediated by the DDAH-ADMA-NO pathway.

Shichimotsukokato (SKT) is a Kampo formula, comprising astragalus root, phellodendron bark, rehmannia root, peony root, cnidium rhizome, Japanese angelica root, and uncaria hook. It is prescribed to hypertensive patients who complain of a sensation of a rush of blood to the head, shoulder stiffness, tinnitus, and dull headache. We investigated the effects of SKT on renal hypertension in Wistar rats subjected to a 5/6 nephrectomy (Nx). Systolic blood pressure (SBP) increased markedly after surgery and remained high in the Nx rats. Oral treatment of SKT extract at dosages of 0.75 and 1.5 g/kg/day (corresponding to 5- and 10-fold human dosages, respectively) caused a significant suppression of the increase in SBP in Nx rats. Plasma concentrations of nitric oxide (NO) were marginally lower and asymmetric dimethylarginine (ADMA) significantly higher in the Nx rats than in sham-operated rats. SKT administration caused a significant counteraction of these changes. Finally, we evaluated the levels of protein methyltransferase (PRMT), an enzyme that catalyzes the production of ADMA, and the levels of dimethylarginine dimethylaminohydrolase (DDAH), an enzyme involved in the degradation of ADMA, in the remnant kidney. Neither Nx nor SKT treatment affected PRMT-1 or DDAH-1 levels. DDAH-2 levels were reduced significantly in the Nx rats compared with the sham-operated rats. SKT treatment significantly ameliorated this decrease in the DDAH-2 levels. It is considered that SKT reduced blood pressure in the renal hypertension rat model, mediated, at least partially, by the DDAH-ADMA-NO pathway.

Secolignans with antiangiogenic activities from Peperomia dindygulensis.

Two new secolignans, peperomins G and H (1 and 2, resp.), were isolated from the whole plant of Peperomia dindygulensis, together with five known secolignans, peperomin A (3), peperomin E (4), peperomin B (5), 2,3-trans-2-methyl-3-{(3-hydroxy-4,5-dimethoxyphenyl)[5-methoxy-3,4-(methylenedioxy)phenyl]methyl}butyrolactone (6), 2,3-cis-2-(hydroxymethyl)-3-{bis[5-methoxy-3,4-(methylenedioxy)phenyl]methyl}butyrolactone (7). Their structures and configurations were elucidated by spectroscopic methods including 2D-NMR techniques. Antiangiogenic effects of all compounds were evaluated using human umbilical vein endothelial cells (HUVEC) proliferation and tube-formation tests, with compounds 4 and 5 being active in the bioassay. Compounds 4 and 5 induced obvious cell toxicity to HUVEC with IC(50) values of 1.64±0.19 and 8.44±0.4 µM, respectively. Compounds 4 and 5 also exhibited significant HUVEC tube formation-inhibiting activity with IC(50) values of 3.13±0.09 and 6.24±0.12 µM, respectively.