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Pan-Pan Wu,1–3 Hao He,1,3 W David Hong,1,3–5 Tong-Rong Wu,1,3 Gui-Ying Huang,2 Ying-Ying Zhong,2 Bo-Rong Tu,1,3 Min Gao,1,3 Jun Zhou,1,3 Su-Qing Zhao,1–3 Dong-Li Li,1,3 Xue-Tao Xu,1,3 Zhao-Jun Sheng,1,3 Stephen A Ward,4 Paul M O’Neill,5 Kun Zhang1–3 1Faculty of Chemical and Environmental Engineering, Wuyi University, Jiangmen, China; 2Department of Pharmaceutical Engineering, Faculty of Chemical Engineering and Light Industry, Guangdong University of Technology, Guangzhou, China; 3International Healthcare Innovation Institute (Jiangmen), Jiangmen, China; 4Research Centre for Drugs and Diagnostics, Liverpool School of Tropical Medicine, Liverpool, UK; 5Department of Chemistry, University of Liverpool, UK Background: Fusidic acid (FA) (WU-FA-00) is the only commercially available antimicrobial from the fusidane family that has a narrow spectrum of activity against Gram-positive bacteria.Methods: Herein, the hydrogenation derivative (WU-FA-01) of FA was prepared and both compounds were examined against a panel of six bacterial strains. In addition, their anti-inflammatory properties were evaluated using a 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse ear edema model.Results: The results of the antimicrobial assay revealed that both WU-FA-00 and WU-FA-01 displayed a high level of antimicrobial activity against Gram-positive strains. Moreover, killing kinetic studies were performed and the results were in accordance with the minimum inhibitory concentration and minimum bactericidal concentration results. We also demonstrated that the topical application of WU-FA-00 and WU-FA-01 effectively decreased TPA-induced ear edema in a dose-dependent manner. This inhibitory effect was associated with the inhibition of TPA‑induced upregulation of proinflammatory cytokines IL-1β, TNF-α, and COX-2. WU-FA-01 significantly suppressed the expression levels of p65, IκB-α, and p-IκB-α in the TPA-induced mouse ear model. Conclusion: Overall, our results showed that WU-FA-00 and WU-FA-01 not only had effective antimicrobial activities in vitro, especially to the Gram-positive bacteria, but also possessed strong anti-inflammatory effects in vivo. These results provide a scientific basis for developing FA derivatives as antimicrobial and anti-inflammatory agents. Keywords: fusidic acid, derivative, antimicrobial, anti-inflammatory

Background: Post-viral olfactory dysfunction (PVOD) is the common symptoms of long COVID, lacking of effective treatments. Traditional Chinese medicine (TCM) is claimed to be effective in treating olfactory dysfunction, but the evidence has not yet been critically appraised. We conducted a systematic review to evaluate the effectiveness and safety of TCM for PVOD., Methods: We searched eight databases to identified clinical controlled studies about TCM for PVOD. The Cochrane risk of bias tools and GRADE were used to evaluate the quality of evidence. Risk ratio (RR), mean differences (MD), and 95 % confidence interval (CI), were used for effect estimation and RevMan 5.4.1 was used for data analysis., Results: Six randomized controlled trials (RCTs) (545 participants), two non-randomized controlled trials (non-RCTs) (112 participants), and one retrospective cohort study (30 participants) were included. The overall quality of included studies was low. Acupuncture ( n = 8) and acupoint injection ( n = 3) were the mainly used TCM therapies. Five RCTs showed a better effect in TCM group. Four trials used acupuncture, and three trials used acupoint injection. The results of two non-RCTs and one cohort study were not statistically significant. Two trials reported mild to moderate adverse events (pain and brief syncope caused by acupuncture or acupoint injection)., Conclusions: Limited evidence focus on acupuncture and acupoint injection for PVOD and suggests that acupuncture and acupoint injection may be effective in improving PVOD. More well-designed trials should focus on acupuncture to confirm the benefit., Protocol Registration: The protocol of this review was registered at PROSPERO: CRD42022366776., Competing Interests: The author JPL is senior editorial board member of the journal, but none of the authors has any conflict of interest regarding this study., (© 2024 Korea Institute of Oriental Medicine. Published by Elsevier B.V.)

In order to develop original water soluble antagonists of X-linked inhibitor of apoptosis protein (XIAP), a novel bicyclic scaffold was designed based on 8,5-fused bicyclic lactam. During its preparation, a spontaneous rearrangement from 8,5- to 7,5-fused bicyclic lactam was observed and confirmed by MS and NMR analyses, in particular the HMBC spectra. DFT calculations were performed to understand the corresponding mechanism. It was finally prevented through changing the reaction order in the synthesis route and a Smac mimetic with this core structure, ZJ-1 was successfully obtained. The structure of this new bicyclic scaffold was well confirmed by HRMS and NMR ( 1 H, 13 C, NOESY) analyses. ZJ-1 presented in addition a binding affinity to XIAP-BIR3, nearly 6 times better than that of AVPI, similar to the reported SM-128 in an in vitro fluorescence polarization (FP) assay. This preliminary result suggests that this new bicyclic scaffold could be very attractive in the development of novel anticancer agents targeting XIAP.

2-Substituted-1,4,5,6-tetrahydrocyclopenta[b]pyrrole, a key structural moiety exiting in many bioactive molecules, has been shown to have excellent selective activity on COX-2. In the present study, the anti-inflammatory activity and the underlying molecular mechanism of 2-substituted-1,4,5,6-tetrahydrocyclopenta[b]pyrrole on skin inflammation were assessed by 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced skin inflammation in mice. Most of the compounds showed anti-inflammatory activity on TPA-induced skin inflammation. The anti-inflammatory activity of compound 4 showed higher anti-inflammatory activity than celecoxib (3.2-fold). Compound 4 pretreatment resulted in markedly suppression of TPA-induced IL-1β, IL-6, TNF-α, and COX-2, respectively. Furthermore, the mechanical study indicated that the anti-inflammatory activity of compound 4 was associated with its ability to inhibit activation of factor kappa-κB (NF-κB) by blocking IκB kinase (IKK) activities. Accordingly, compound 4 could be used as a potential anti-inflammatory agent for skin inflammation., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Liver cancer is one of the most common cancers in the world. Macrothele raven venom, a complicated mixture of neurotoxic peptides, proteins and low molecular weight material, has antitumor properties, but its mechanism of action is unknown. Moderate exercise has been shown to shrink tumors and cause a remarkable reduction in the tumor growth rate. In this study, we examined the antitumor effect of Macrothele raven venom in combination with exercise on tumor-bearing mice. Our results demonstrate that aerobic exercise in combination with venom administered at different doses was much more effective in a mouse H22 hepatoma model compared to separate administration of the 2 treatments. The underlying mechanism of this effect may be related to the expression of various tumor suppressor factors., (© Georg Thieme Verlag KG Stuttgart · New York.)

Background: It is well-documented that both chemokine (C-C motif) ligand 19 (CCL19) and 21 (CCL21) mediate cell migration and angiogenesis in many diseases. However, these ligands' precise pathological role in ankylosing spondylitis (AS) has not been elucidated. The objective of this study was to examine the expression of CCL19 and CCL21 (CCL19/CCL21) in AS hip ligament tissue (LT) and determine their pathological functions., Methods: The expression levels of CCL19, CCL21 and their receptor CCR7 in AS (n = 31) and osteoarthritis (OA, n = 21) LT were analyzed via real-time polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC). The expression of CCL19, CCL21 and CCR7 in AS ligament fibroblasts was also detected. The proliferation of ligament fibroblasts was measured via a cell counting kit-8 (CCK8) assay after exogenous CCL19/CCL21 treatment. Additionally, the role of CCL19/CCL21 in osteogenesis was evaluated via RT-PCR and enzyme-linked immunosorbent assay (ELISA) in individual AS fibroblast cultures. Furthermore, the expression of the bone markers alkaline phosphatase (ALP), osteocalcin (OCN), collagenase I (COL1), integrin-binding sialoprotein (IBSP) and the key regulators runt-related transcription factor-2 (Runx-2) and osterix were investigated. Moreover, the CCL19/CCL21 levels in serum and LT were measured via ELISA., Results: The mRNA levels of CCL19/CCL21 in AS hip LT were significantly higher than that in OA LT, and IHC analysis revealed a similar result. Exogenous CCL19/CCL21 treatment did not affect the proliferation of ligament fibroblasts but significantly up-regulated the expression of bone markers, including ALP and OCN, and the key regulators Runx-2 and osterix. In addition, the serum levels of CCL19/CCL21 were apparently elevated in AS patients compared to healthy controls (HC), and the expression of the two chemokines correlated significantly in AS patients., Conclusions: CCL19 and CCL21, two chemokines displaying significantly associated expression in serum, indicating a synergistic effect on AS pathogenesis, may function as promoters of ligament ossification in AS patients.

Zinc finger protein 191, ZNF24 and Zfp191 in both humans and mice belong to the SCAN domain subfamily of Krüppel-like zinc finger transcription factors. Previous studies have suggested that Zfp191 is a pleiotropic factor involved in embryonic development, hematopoiesis and tumorigenesis. However, little is known about its target genes or its role in other physiological and pathological processes. We have identified the putative target genes of Zfp191, using an in silico genome-wide scan. Three hundred and fifty-five putative target genes were identified, which were enriched into the pathways of immune response according to the pathway analysis. These targets indicated that Zfp191 may function as a mediator of the immune response. This was verified in mice heterozygous for Zfp191 (Zfp191(+/-)) using a lipopolysaccharide (LPS)-induced endotoxic shock model. After LPS injection, Zfp191(+/-) mice produced significantly less IL-1β and IL-6 compared to wild-type mice and were resistant to LPS-induced endotoxic shock. The loss of Zfp191 may suppress systemic inflammation by reducing these cytokine levels during LPS-induced endotoxic shock.

gamma-Aminobutyric acid (GABA) transporters play a key role in the regulation of GABA neurotransmission. We reported previously that overexpression of the GABA transporter subtype 1 (GAT1), the major form of the GABA transporter in the CNS, led to hyperalgesia in mice. In the present study, nociceptive responses of GAT1-knockout mice (GAT1(-/-)) were compared with those of heterozygous (GAT(+/-)) and wild-type (GAT(+/+)) mice by four conventional pain models (tail-immersion test, hot-plate test, acetic acid-induced abdominal constriction test, and formalin test). In addition, the analgesic effects of two GAT1-selective inhibitors, NO-711 and tiagabine, were examined in all three genotypes using the same four models. Our data demonstrated that GAT1 deficiency because of genetic knockout or acute blockade by selective inhibitors leads to hypoalgesia in mice. These results confirmed the crucial role of GAT1 in the regulation of nociceptive threshold and suggested that GAT1 inhibitors have the potential for clinical use in pain therapy., ((c) 2007 Wiley-Liss, Inc.)

Gamma-aminobutyric acid (GABA) transporter subtype 1 (GAT1), which transports extracellular GABA into presynaptic neurons, plays an important regulatory role in the function of GABAergic systems. However, the contributions of the GAT1 in regulating mental status are not fully understood. In this paper, we observed the behavioral alterations of GAT1 knockout (GAT1(-/-)) mice using several depression- and anxiety-related models (eg, the forced-swimming test and the tail-suspension test for testing depression-related behaviors; the open-field test, the dark-light exploration test, the emergence test, and the elevated plus maze (EPM) test for anxiety-related behaviors). Here we found that GAT1(-/-) mice showed a lower level of depression- and anxiety-like behaviors in comparison to wild-type mice. Furthermore, GAT1(-/-) mice exhibited measurable insensitivity to selected antidepressants and anxiolytics such as fluoxetine, amitriptyline, buspirone, diazepam, and tiagabine in the tail-suspension test and/or the EPM test. Moreover, the basal level of corticosterone was found to be significantly lower in GAT1(-/-) mice. These results showed that the absence of GAT1 affects mental status through enhancing the GABAergic system, as well as modifying the serotonergic system and the hypothalamic-pituitary-adrenal (HPA) activity in mice.

Dysregulation of the brain GABAergic system has been implicated in the pathophysiology of violence and aggression. As a key regulator of central GABAergic activity, dysfunction of the GABA transporter subtype 1 (GAT1) represents a potential mechanism mediating pathologic aggression. We provide evidence that GAT1-/- mice and GAT1+/- mice exhibit lower aggressive behavior both in home cage resident-intruder test and neutral arena resident-intruder test, compared to wild-type mice (GAT1+/+). The pharmacologic effects of the GAT1 inhibitor, tiagabine and the GABA(A) receptor antagonist, bicuculline have been assessed in GAT1+/+ mice: tiagabine inhibits attacks but bicuculline induces attacks. Compared to GAT1+/- and +/+ mice, the GAT1-/- mice displayed a normal circadian pattern of home cage activity, but more activity overall. Meanwhile, reduced testosterone concentration was found in GAT1-/- mice compared to GAT1+/+ mice but not in GAT1+/+ mice treated with tiagabine, suggesting that testosterone is not directly involved in GAT1 mediated aggressive behavior regulation. These results showed that GAT1 is an important target involved in the regulation of aggressive behavior in mice, and long-term dysfunction of GAT1 may also result in the alteration of testosterone secretion.

It is widely accepted that the GABAergic system plays an important role in the action of ethanol in vivo. GABA transporter subtype 1 (GAT1) constructs high affinity reuptake sites in the CNS and regulates GABAergic transmissions. In this study, mice lacking the GAT1 were developed by homologous recombination. Both hetero- and homozygous GAT1 mutant mice were tested for ethanol, saccharin or quinine consumption, ethanol-conditioned place preference, ethanol-conditioned taste aversion, ethanol-simulated motor activity, and ethanol-induced sedation/hypnosis. The GAT1(-/-) mice showed decreased ethanol aversion and ethanol reward, and insensitivity to both the sedative/hypnotic and the motor stimulant effects of ethanol, along with increased avoidance of quinine preference and consumption. GAT1(+/-) mice showed significantly increased consumption of ethanol and saccharin, however, enhanced the rewarding and preference effect of ethanol, increased avoidance of quinine, and higher sensitivity to the motor stimulant effect of ethanol. These results demonstrate that GAT1, perhaps in a bi-directional way, modulates some behavioral effects of ethanol. The GAT1 mutant mice provided us a very useful model to investigate the mechanisms of ethanol action in vivo.

Objective: To investigate the effects of chitosan membrane on tendon adhesion and healing and obtain experimental data for clinical use in preventing postoperative tendon adhesion., Methods: The long flexor tendon of 55 adult legborn hens were cut and sutured encapsulated by chitosan membrane. Movement and anti-tension capability of tendon were assessed at 2, 4, 6, 8 and 10 weeks after operation by SWD-10 type tendon stretcher. Tendon healing and adhesion were observed with light microscope., Results: Tendon healing could be effected by chitosan membrane within 4 weeks, and tendon anti-tension strength was increased after 4 weeks., Conclusion: Chitiosan membrane possesses the following characteristics: no side effects, good permeability, resolvable, absorbable and selective inhibition the growth of fibroblast. It is a desirable biological material to prevent tendon adhesion.

We describe a case of human T-lymphotropic virus type I associated myelopathy in a 50-year old woman in Nigeria. The patient presented with progressive loss of tone to the two lower limbs and later inability to walk. The HTLV-I antibody presence in the plasma collected from the patient was repeatedly detected by enzyme immunoassays (Abbott HTLV-I EIA and Coulter SELECT-HTLV I/II) and confirmed by Western blot technique. In addition, HTLV-I DNA was amplified from the genomic DNA isolated from the peripheral blood mononuclear cells of the patient by the polymerase chain reaction technique. This finding is significant being the first report of association of HTLV-I with myelopathy in Nigeria.

Two transposable elements, IS5376 and IS5377, were identified in the thermophile Bacillus stearothermophilus CU21 based upon the following criteria: (1) both were found to appear on different plasmids introduced into the same host CU21; (2) signals of homology were found between the genomic DNA of CU21 and each of them; (3) different numbers of Southern hybridization bands were found for the genomic DNA of different strains of B. stearothermophilus; and (4) characteristic inverted repeats at both ends and direct repeats of the target DNA adjacent to them were found for both IS5376 and IS5377. Two open reading frames (ORFs) were detected for IS5376 and one for IS5377. The putative coding products of the ORFs are homologous to those of known ISs from mesophiles and are considered to be transposases. The results of analyses of nucleotide sequence and the deduced amino acid sequence suggest that IS5376 is a member of the IS21 family and that IS5377 is a member of the IS4 family.

Two genes of the meta pathway of phenol degradation were cloned from a phenol-utilizing strain of Bacillus stearothermophilus and were mapped by subcloning and by use of a Tn5 insertion mutation. They code for phenol hydroxylase and catechol 2,3-dioxygenase, respectively. The gene encoding catechol 2,3-dioxygenase, which is more thermostable than catechol 2,3-dioxygenase encoded by the other gene, shares rather limited homology with that from Pseudomonas putida.

Plasmid pUC13 was used to clone DNA fragments of known sites from the chromosome of Escherichia coli. Each chimeric plasmid was introduced individually into the same dnaA46 mutant strain LC381 and suppressive integration (Sin) strains were selected. By means of cotransduction the null mutation recA56 was then introduced into each Sin strain and growth of each recA56 derivative at 42 degrees C was scored. Strains that failed to grow at 42 degrees C depended upon the recA gene for replication. Three factors were shown to limit the viability of LC381 harboring different chimeric plasmids and affect the degree of recA gene dependence of chromosome replication in the Sin strains at 42 degrees C. It is suggested that these three constraints are the consequence of the organization of the E. coli chromosome, particularly the unique ability of terC to retard the progression of replication forks. Two classes of hypotheses concerning the function of the recA gene are considered.

We have reported in our previous paper that replication of E. coli chromosome initiated by the integrated F' plasmid depends on the recA gene. Here we report on work dealing with the role played by recA, recB, recC, lexA alleles in chromosome replication. Our results show that the recA gene takes part in chromosome replication through homologous recombination rather than SOS pathway and that Chi hot spot is not concerned with. They also show that the ATP-dependent dsDNA exonuclease activity of the RecBC enzyme has nothing to do with the recA gene in chromosome replication.

mini-F plasmids carrying particular chromosomal fragments were constructed. Through homologous recombination they were expected to integrate onto predetermined sites of the chromosome of dnaA46 strain LC381. Chromosome replication of suppressive integration (Sin) strains were recA dependent or independent according to the site of integration of the plasmid: chromosome replication of those with the plasmid integrated close to oriC were found to be recA dependent at 40 degrees C; those with the plasmid integrated between oriC and terC were recA dependent on rich medium but independent on minimal medium; those with the plasmid integrated close to terC were recA dependent even on minimal medium.

E. coli dnaA46 fails to grow and form colonies at 40 degrees C; integrated with plasmid enables it to grow at 40 degrees C or 42 degrees C. In a previous paper we reported that the replication of chromosome initiated by the integrated F' plasmid was recA gene dependent. In this paper we report further the behavior of 5 plasmids and 2 phages as to their dependence of recA gene in the integrated state. It was demonstrated that the dependence or independence of recA gene is not related to the direction of replication of the plasmids or phages in the free state. The hypothesis that the function of recA gene is to convert the unidirectionally replicating machinery in the free state to the bidirectionally replicating one in the integrated state is refuted accordingly.

pFDX1 is a recombinant plasmid which carries a foreign gene xylE. By selecting for kanamycin-resistant mutants of Bacillus stearothermophilus CU21(pFDX1) at higher temperature, a variant strain CU21-163 was obtained. This strain harbors a mutant plasmid pFDX163, which was formed by insertion of a 2.0kb H-fragment from the CU21 genome onto the plasmid pFDX1. pFDX163 was supposed to be integrated into the CU21 chromosome via homologous recombination of H-fragments. The CU21-163 strain consists of two cell types, i.e. y-cell and w-cell. The expression level of xylE gene in the former is higher than that in the latter. The progeny of a y-cell always contains some w-cells, while that of a w-cell contains y-cells. This is supposed to be due to a phase variation of CU21-163. Analysis on the amount of free and integrated plasmid DNA in different DNA samples of CU21-163 cells allows us to draw the conclusion that there are both free and integrated plasmids in the y-cells, whereas only integrated ones in the w-cells.

We have reported the dependence of recA gene for the replication of chromosome of E. coli initiated by the F' plasmid but not the F plasmid. Mini-F plasmids with ISI sequence and origin of replication from F and F' plasmid have been constructed. 20% of the integrative suppression strains of these mini-F plasmids were found to be recA dependent, irrespective of the origin of replication (F or F' plasmid) and the direction of replication (uni- or bidirectional). The reported experimental results tend to suggest that the site of integration is of primary importance in the dependence vs independence of recA gene for the replication of the chromosome initiated by the integrated plasmid.

Using the gene expression vector pFDC11 for Bacillus stearothermophilus CU21, a recombinant plasmid pFDX1 was constructed, which carries the Pseudomonas-derived xylE gene coding for Catechol 2,3-dioxygenase (CatO2ase). CatO2ase activity can be detected from CU21 (pFDX1) cells grown at 48 degrees C. This result indicates that the promoterless xylE gene can be expressed in a thermophilic host under the direction of a promoter from a thermophilic bacterium. By selecting Kmr mutants at elevated growth temperature, a segregant CU21-161 was obtained, the xylE gene expression of which at 55 degrees C and 60 degrees C was much higher than that of the parental strain. A method for CatO2ase assay with suspension of intact cells was also reported in this paper.

A low copy number mutant of ColE1-like plasmid of Escherichia coli was reported in this paper. The recombinant plasmid pPGVT3 derived from the vector pUC4 was unstable in the E. coli host strain DF2145. No transformant could be obtained at 40 degrees C when DF2145 was transformed with pPGVT3. By in vitro mutagenesis of pPGVT3 plasmid DNA with hydroxylamine which induces point mutations, a mutant plasmid pPGVT3HA was obtained, which was stable in DE2145. The mutation site was localized within the pUC moiety of pPGVT3HA. The copy number of the mutated pUC vector and its derivatives were found to be reduced. The effect of low copy number mutation on the stability of the recombinant plasmids was discussed.

Mutant strain dnaA46 of Escherichia coli can be integratively suppressed by the F' plasmid. Upon introducing the recA56 mutation through transduction the suppressive integration strain (Sin) becomes unable to grow at 40 degrees C. By means of experiments of marker transfer, acridine orange sensitivity test, F' curing and mini-chromosome transformation it is concluded that the F' plasmid is always in an integrated state in the Sin strains and that the initiation of the replication of the bacterial chromosome is carried on by the integrated F' plasmid. The biosynthesis of DNA and protein of the Sin recA+ and Sin recA- strains at different temperatures were compared. It is concluded from the experimental results that the recA gene functions at the level of DNA replication. The recA gene is known to be the key gene in DNA recombination and SOS repair of DNA damage. The works reported here throw some light on the understanding of the function of the recA gene.