Corydecusines A-H, new phthalideisoquinoline hemicetal alkaloids from the bulbs of Corydalis decumbens inhibit Tau pathology by activating autophagy mediated by AMPK-ULK1 pathway.

Twelve phthalideisoquinoline hemiacetal alkaloids including eight new ones (1-8) and one natural alkaloid characterized by an aziridine moiety with unassigned NMR data (9), were isolated and identified from the bulbs of Corydalis decumbens. Their structures were established by comprehensive analyses of HRESIMS, NMR, X-ray crystallography, and ECD analyses. The unambiguously established structures of the phthalideisoquinoline hemiacetal alkaloids indicated that the absolute configurations of C-1, C-9, and C-7' were confusable only relied on coupling constants. A summary of their ECD spectra was concluded and provided an insight for C-1, C-9, and C-7' absolute configuration assignment. These new compounds were evaluated to induce autophagy flux through flow cytometry analysis. Moreover, compounds 2 and 6 could significantly induce autophagy and inhibit Tau pathology by AMPK-ULK1 pathway activation, which provided an avenue for anti-AD lead compounds discovery.

New Monoterpenoid Indole Alkaloids from Tabernaemontana crassa Inhibit ß-Amyloid42 Production and Phospho-Tau (Thr217).

Eleven monoterpenoid indole alkaloids, including three new ones, tabercrassines A-C (1-3), were isolated from the seeds of Tabernaemontana crassa. Tabercrassine A (1) is an ibogan-ibogan-type bisindole alkaloid which is formed by the polymerization of two classic ibogan-type monomers through a C3 unit aliphatic chain. Their structures were established by extensive analysis of HRESIMS, NMR, and ECD spectra. Cellular assays showed that alkaloids 1-3 all reduce Aß42 production and inhibit phospho-tau (Thr217), a new biomarker of Alzheimer's disease [AD] associated with BACE1-, NCSTN-, GSK3ß-, and CDK5-mediated pathways, suggesting these alkaloids' potential against AD.

Alkaloids from Tabernaemontana divaricata combined with fluconazole to overcome fluconazole resistance in Candida albicans.

Nineteen indole alkaloids including eleven new ones, taberdines A-K (1-11), were isolated from Tabernaemontana divaricata. Their structures were assigned by MS, NMR, single crystal X-ray diffractions, and ECD analyses. Alkaloid 1 is an aspidosperma-type monoterpenoid indole alkaloid and possesses a rearranged pyrrolidine moiety due to C-3 degradation, and 4 has a rare 1,3-oxazolidine moiety within iboga-type alkaloids. Alkaloids 2, 4, 6, and 11-19 combined with 5 µg/mL fluconazole exhibited significant activity to reverse fluconazole resistance in Candida albicans strains while no one used alone showed any activities against the resistant strain.

Cytotoxic Monoterpenoid Indole Alkaloids from Tabernaemontana corymbosa as Potent Autophagy Inhibitors by the Attenuation of Lysosomal Acidification.

Twenty-six alkaloids, including the new taberines A-I (1-9), were obtained from Tabernaemontana corymbosa. The structures and absolute configurations were elucidated via MS, NMR, and ECD spectroscopic data analyses. Alkaloids 1-4 are new vobasinyl-ibogan alkaloids, and 1 is characterized by an unusual 1,3-oxazinane moiety. Alkaloids 4 and 16 exhibited moderate cytotoxic potency against various human cancer cell lines, while 4, 10, 11, 13, 14, and 16 showed attenuation of lysosomal acidification activity (EC50: 12.9-29.8 µM), thereby inhibiting autophagic flux.

Antiproliferative Aspidosperma-Type Monoterpenoid Indole Alkaloids from Bousigonia mekongensis Inhibit Tubulin Polymerization.

Monoterpenoid indole alkaloids are structurally diverse natural products found in plants of the family Apocynaceae. Among them, vincristine and its derivatives are well known for their anticancer activity. Bousigonia mekongensis, a species in this family, contains various monoterpenoid indole alkaloids. In the current study, fourteen known aspidosperma-type monoterpenoid indole alkaloids (1⁻14) were isolated and identified from a methanol extract of the twigs and leaves of B. mekongensis for the first time. Among them, compounds 3, 6, 9, and 13 exhibited similar antiproliferative activity spectra against A549, KB, and multidrug-resistant (MDR) KB subline KB-VIN cells with IC50 values ranging from 0.5⁻0.9 µM. The above alkaloids efficiently induced cell cycle arrest at the G2/M phase by inhibiting tubulin polymerization as well as mitotic bipolar spindle formation. Computer modeling studies indicated that compound 7 likely forms a hydrogen bond (H-bond) with α- or ß-tubulin at the colchicine site. Evaluation of the antiproliferative effects and SAR analysis suggested that a 14,15-double bond or 3α-acetonyl group is critical for enhanced antiproliferative activity. Mechanism of action studies demonstrated for the first time that compounds 3, 4, 6, 7, and 13 efficiently induce cell cycle arrest at G2/M by inhibiting tubulin polymerization by binding to the colchicine site.

Taburnaemines A-I, Cytotoxic Vobasinyl-Iboga-Type Bisindole Alkaloids from Tabernaemontana corymbosa.

Nineteen vobasinyl-ibogan-type bisindole alkaloids, including nine new compounds, taburnaemines A-I (1-9), were isolated from the twigs and leaves of Tabernaemontana corymbosa. The structures and absolute configurations of the new alkaloids were determined by a combination of MS, NMR, and ECD analyses. Alkaloids 1-5 contain a rare 1,3-oxazinane moiety in the vobasinyl unit, while 6 has an uncommon 1,3-oxazolidine moiety in the iboga unit. The absolute configurations of alkaloid 1 and the known alkaloid tabernaecorymbosine A (10) were confirmed by single-crystal X-ray diffraction analysis. All of the bisindole alkaloids, except 2 and 16'-decarbomethoxytabernaecorymbosine A (14), showed antiproliferative activity (IC50 2.6-9.8 µM) against several human cancer cell lines, including A-549, MDA-MB-231, MCF-7, KB, and P-glycoprotein-overexpressing multidrug-resistant KB cells. The preliminary structure-activity relationship correlations are also discussed.

Hydrogel Microneedle Patches Loaded with Stem Cell Mitochondria-Enriched Microvesicles Boost the Chronic Wound Healing.

Rescuing or compensating mitochondrial function represents a promising therapeutic avenue for radiation-induced chronic wounds. Adult stem cell efficacies are primarily dependent on the paracrine secretion of mitochondria-containing extracellular vesicles (EVs). However, effective therapeutic strategies addressing the quantity of mitochondria and mitochondria-delivery system are lacking. Thus, in this study, we aimed to design an effective hydrogel microneedle patch (MNP) loaded with stem cell-derived mitochondria-rich EVs to gradually release and deliver mitochondria into the wound tissues and boost wound healing. We, first, used metformin to enhance mitochondrial biogenesis and thereby increasing the secretion of mitochondria-containing EVs (termed "Met-EVs") in adipose-derived stem cells. To verify the therapeutic effects of Met-EVs, we established an in vitro and an in vivo model of X-ray-induced mitochondrial dysfunction. The Met-EVs ameliorated the mitochondrial dysfunction by rescuing mitochondrial membrane potential, increasing adenosine 5'-triphosphate levels, and decreasing reactive oxygen species production by transferring active mitochondria. To sustain the release of EVs into damaged tissues, we constructed a Met-EVs@Decellularized Adipose Matrix (DAM)/Hyaluronic Acid Methacrylic Acid (HAMA)-MNP. Met-EVs@DAM/HAMA-MNP can load and gradually release Met-EVs and their contained mitochondria into wound tissues to alleviate mitochondrial dysfunction. Moreover, we found Met-EVs@DAM/HAMA-MNP can markedly promote macrophage polarization toward the M2 subtype with anti-inflammatory and regenerative functions, which can, in turn, enhance the healing process in mice with skin wounds combined radiation injuries. Collectively, we successfully fabricated a delivery system for EVs, Met-EVs@DAM/HAMA-MNP, to effectively deliver stem cell-derived mitochondria-rich EVs. The effectiveness of this system has been demonstrated, holding great potential for chronic wound treatments in clinic.

Ox-LDL-induced TGF-ß1 production in human alveolar epithelial cells: involvement of the Ras/ERK/PLTP pathway.

Oxidized-low density lipoprotein (Ox-LDL) has been shown to play an important role in impaired surfactant metabolism and transforming growth factor-ß1 (TGF-ß1) is a critical mediator in the pathogenesis of acute lung injury (ALI)/acute respiratory distress syndrome (ARDS). In this study, we investigated whether Ox-LDL can induce TGF-ß1 protein production, and if so, how it achieves this induction in human alveolar epithelial cells (A549). We show here that Ox-LDL not only caused a dose- and time-dependent up-regulation of TGF-ß1 production, but also increased Smad3 phosphorylation, Ras/extracellular signal-regulated kinase (ERK) activity and phospholipid transfer protein (PLTP) expression in A549 cells. The inhibition of Ras/ERK activity with specific inhibitors significantly suppressed Ox-LDL-induced TGF-ß1 production, Smad3 phosphorylation and PLTP expression. Furthermore, treatment of cells with PLTP siRNA suppressed both TGF-ß1 release and Smad3 activation induced by Ox-LDL, but not the activation of Ras/ERK cascade. Taken together, we provide evidences that induction of TGF-ß1 production and Smad3 phosphorylation by Ox-LDL is mediated by Ras/ERK/PLTP pathway in human alveolar epithelial cells.

Monoterpenoid indole alkaloid dimers from Kopsia arborea inhibit cyclin-dependent kinase 5 and tau phosphorylation.

Three undescribed monoterpenoid indole alkaloid dimers (kopoffines A-C, which are connected via a methylene unit) and with nine known alkaloids were isolated and identified from the fruits of Kopsia arborea Blume. Their structures, including their absolute configurations, were established by HRESIMS, NMR, single-crystal X-ray diffraction, and ECD analyses. Kopoffines A-C showed significant inhibition against cyclin-dependent kinase 5 (IC50: 0.34-2.18 µM). Western blotting analyses showed that kopoffines A-C significantly decreased the protein levels of CDK5 and phospho-CDK5 (Tyr15) (pCDK5) at concentrations of 2.5 and 10 µM. The levels of phospho-Tau (Thr217) (pTau217, a new biomarker of AD), and phospho-Tau (Ser396) (pTau396), which play major roles in the formation of neurofibrillary tangles , were decreased by the kopoffines A-C treatment. Molecular docking studies indicated that kopoffines A-C could form stable interactions with CDK5.

Combined flaps for reconstructing wide-range facial defects.

BACKGROUND: This report aims to describe the technique of using combined flaps for reconstruction of wide-range facial defects. METHODS: Two or more flaps were combined to repair wide-range facial wounds, complying with the principle of facial aesthetic subunits to maintain the inherent structural characteristics of the face, to design incision lines conforming to the natural facial contour lines (e.g., nasolabial fold, sulcus alaris, nasomaxillary suture, palpebral margin, vermilion border, hairline) or wrinkles, and to leave hidden incision lines if possible. There were 18 cases with excessively large facial defects requiring the use of a graft with local flaps. RESULTS: From April 1996 to May 2008, 124 patients had their wide-range facial defects repaired by combined flaps. The size of the defects ranged in size from 2.5 × 4 to 9 × 10 cm. The healing for all the patients was uneventful, and the flap survived completely without complications. After an average of 6 months (range, 1-48 months), 87 of the patients were available for follow-up evaluation. There was no recurrence of tumor. The repaired tissues were well matched with surrounding tissue in color, texture, and contour. Incision lines were sheltered, and there were no anatomic deformities of the eyelid, upper lip, or nasal ala. CONCLUSIONS: Based on the facial aesthetic subregion principle, combined flaps were used to reconstruct wide-range facial defects effectively, obtaining good color, contour, and texture match with the surrounding skin. All in all, the cosmetic results were satisfactory.

Simultaneous deposition of tannic acid and poly(ethylene glycol) to construct the antifouling polymeric coating on Titanium surface.

Titanium (Ti) and its alloys are primarily explored to produce biomedical implants owing to their improved mechanical stability, corrosion resistance, low density, and good biocompatibility. Despite, Ti substrate surfaces are easily contaminated by plasma proteins and bacteria. Herein, a simple one-step process for the simultaneous deposition of a polyphenol tannic acid (TA) and four-armed poly(ethylene glycol) (PEG10k-4-OH) on the Ti substrate (Ti-TA/PEG) surface was described. Additionally, a two-step process has been employed to fabricate the Ti-TA-PEG surface via successive deposition of TA and PEG10k-4-OH for comparison. The resultant Ti-TA/PEG surface prepared by simultaneous deposition of TA and PEG10k-4-OH exhibits higher coating thickness and better surface coverage than the Ti-TA-PEG surface. The Ti-TA/PEG and Ti-TA-PEG surfaces could actively inhibit the non-specific adsorption of proteins, suppress the bacterial and platelet adhesion, and prevents biofilm formation. Moreover, the Ti-TA/PEG surface displays a better antifouling performance than the Ti-TA-PEG surface. Thus, the present study demonstrates a simple and convenient approach for constructing polymeric coating with good anti-adhesive properties on the Ti substrate surface.

[Effect of shenqi fuzheng injection combined with chemotherapy in treating colorectal cancer].

OBJECTIVE: To explore the effect and mechanism of Shenqi Fuzheng Injection (SFI) for assisting of chemotherapy in treating colorectal cancer (CRC). METHODS: Forty CRC patients were randomly and equally assigned to two groups, the control group received chemotherapy of FOLFOX protocol and the test group treated with the same chemotherapy combining with SFI. The CD4+ CD25+ regulatory T (Treg) cells, tumor necrosis factor-alpha (TNF-alpha) and interleukin-12 (IL-12) in peripheral blood were determined before and after treatment, and the toxicity of chemotherapy assessed according to the WHO criteria for acute and subacute toxic reaction of anticancer drugs. RESULTS: Two cases in the control group were lost during the observing period. The amount of CD4+ CD25+ Treg cells in peripheral blood in CRC patients was significantly higher than the normal range (P<0.05), which was lowered significantly after treatment in both groups (P<0.05). Levels of TNF-alpha and IL-12 significantly elevated in the test group after treatment but lowered in the control group, showing significant difference between groups (both P<0.05). As compared with the control group, the adverse reaction to the chemotherapy was significantly lessened in the test group (P<0.05). CONCLUSION: Using SFI for assisting chemotherapy could not only improve the immune function of organism to enhance the effect of chemotherapy, but also reduce the adverse reaction of the chemotherapy.

The bilobed flap for reconstruction of distal nasal defect in Asians.

BACKGROUND: The aim of this study was to overcome nasal alar retraction that occurs with the use of the standard Zitelli's bilobed flap for reconstruction of a distal nasal defect. Zitelli's bilobed flap offers the ability to aesthetically reconstruct difficult defects in the distal nasal area. However, the standard Zitelli bilobed flap may sometimes result in nasal alar retraction which is likely to the result of excessive tension upon closure of the primary lobe and primary defect. METHODS: We modified the bilobed flap based on the design of the standard Zitelli bilobed flap. The modification of the bilobed flap was designed to have a primary lobe that was 10% longer than the length of the distal defect edge from the flap's pivot point, and the width of the primary lobe was equal to the width of the defect. The length of the secondary lobe was 130% of the length of the distal defect edge to the flap's pivot point, and the width of the secondary lobe was two-thirds the width of the primary lobe. To make a slight downward displacement of the ipsilateral alar margin, the primary lobe was transposed to fill the primary defects. RESULTS: The results of the reconstruction were satisfactory: there was good nasal contour and appropriate symmetry of the nasal tip and no alar retraction. CONCLUSION: The modification of Zitelli's bilobed flap to have a longer primary lobe, which slightly displaced downward the ipsilateral alar margin intraoperatively, can prevent nasal alar retraction in the reconstruction of distal nasal defects in Asians.

Melocochines A and B, Two Alkaloids from the Fruits of Melodinus cochinchinensis.

Melocochines A (1) and B (2), one pair of epimers with an unprecedented skeleton, were isolated from Melodinus cochinchinensis. Their structures and absolute configurations were established by a combination of MS, NMR, and single-crystal X-ray diffraction analyses. Compounds 1 and 2 represent a class of novel alkaloids, characterized by a rare 1H-benzo[b]azepane ring system within monoterpenoid indole alkaloid categories. Compounds 1 and 2 enhanced lysosomal biogenesis with LysoTracker staining intensities of 139.7% and 119.0%, respectively.

Tabercorymines A and B, Two Vobasinyl-Ibogan-Type Bisindole Alkaloids from Tabernaemontana corymbosa.

Tabercorymines A (1) and B (2), two new vobasinyl-ibogan-type bisindole alkaloids with an unprecedented skeleton, were isolated from Tabernaemontana corymbosa. Their structures were established by a combination of spectroscopic data, chemical transformation, single-crystal X-ray diffraction, and ECD calculation. Compound 1 represents a novel bisindole alkaloid, characterized by a caged heteropentacyclic ring system incorporating an unprecedented C-7/C-20 bond in the vobasinyl unit. Alkaloids 1 and 2 showed potent antiproliferative activity against several human cancer cell lines, including vincristine-resistant KB.

Functional analysis of HBO1 in tumor development and inhibitor screening.

The aim of the present study was to explore the functions of histone acetyltransferase binding to origin recog-nition complex (ORC) 1 (HBO1) during tumor development and to screen for HBO1 inhibitors. The chromatin immuno-precipitation sequencing (ChIP-seq) data of HBO1 in the RKO human colon cancer cell line (GSE33007) were downloaded from the Gene Expression Omnibus (GEO) database. The reads were then mapped back to a reference genome hg19. The PCR duplicate reads were removed by using SAMtools software and the shift was calculated using SPP and MaSC software. The peak calling was carried out using MACS 1.4.0 software. Furthermore, the inhibitors of HBO1 were screened out from the Specs database using Dock 6.6 software. The binding sites of HBO1 were mainly distributed in the intergenic, intronic and 3'-end regions. Further analysis revealed that a total of 9,467 target genes was identified around HBO1 binding sites in the RKO cell lines and those genes mainly participated in the cell cycle, biosynthetic process, as well as other processes. Finally, 5 inhibitors with best binding affinity in the positively charged cavity of HBO1 were screened out: i) 5-[(2-hydroxybenzylidene)amino] -2-(2­{4­[(2­hydroxy-benzylidene)amino]-2-sulfonatophenyl}vinyl)benzenesulfonate, ii) 3-[4-(3-bromo-4-{2-[4-(ethoxycarbonyl)anilino]-2-oxo-ethoxy}-5-methoxybenzylidene)­3­methyl­5­oxo -4,5-dihydro-1H-pyrazol-1-yl]benzoic acid, iii) 4-(4-{3-iodo­5­ methoxy­4-[2-(2-methoxyanilino)-2-oxoethoxy]benzylidene}-3-methyl-5-oxo-4,5-dihydro-1H-pyrazol-1-yl)benzoic acid, iv) 5-chloro-1,3-bis{[3,5,6-trihydroxy-4-(octyloxy)tetrahydro-2H-pyran-2-yl]methyl}-1,3-dihydro-2H-benzimidazol-2-one and v) 4-{[4-(tetradecylamino)-1-naphthyl]diazenyl}benzoic acid. As a whole, in this study, we identified the possible binding sites and biological functions of HBO1. The potential inhibitors of HBO1 were also screened, which prove to be helpful for the inhibition of HBO1 during tumor development.

[Association of surfactant protein D gene polymorphisms at rs3088308 and rs721917 with susceptibility to silicosis].

OBJECTIVE: To investigate the relationship between polymorphisms of surfactant protein D (rs3088308 and rs721917) and the susceptibility to silicosis. METHODS: This case-control study included 125 silicosis patients and 125 individuals exposed to industrial dust but without silicosis (control group), who were strictly matched with the case group for age, gender, work type and cumulative length of dust exposure. The rs3088308 and rs721917 polymorphisms of surfactant protein-D were detected in all the participants using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: The frequencies of T/T, T/A and A/A genotypes of surfactant protein-D rs3088308 locus were 22.2%, 71.2% and 5.6% in the case group, significantly different from the frequencies of 17.6%, 58.4% and 24.0% in the control group, respectively (P<0.05). The frequencies of C/C, C/T and T/T genotypes of rs721917 locus were 17.6%, 56.8% and 25.6% in the case group, similar to the frequencies of 15.2%, 60.0% and 24.8% in the control group, respectively (P>0.05). CONCLUSION: Surfactant protein-D rs3088308 polymorphism is significantly associated with silicosis, and the T allele may be a risk factor for silicosis in individuals exposed to industrial dust.

Chemical Constituents from the Stems of Manihot esculenta.

Two new compounds, maniesculentins A (1) and B (6), together with four known ones were isolated from the stems of Manihot esculenta Crantz. The structures of the new compounds were elucidated by extensive spectroscopic methods including NMR spectroscopy and mass spectrometry. The two new compounds (1, 6) were assayed for antibacterial activity against four tested bacteria lines.

Advances in mechanisms of microglial function on demyelination and remyelination in EAE model / 中国免疫学杂志

Inflammatory cells infiltration and demyelination in the central nervous system (CNS) are the main pathological characteristics of multiple sclerosis(MS),an autoimmune disease in the CNS.Most of the related pathological studies are carried out in the animal model experimental autoimmune encephalomyelitis(EAE).Microglia(MG) are the primary immune effector cells of the CNS and their activation can play complex roles in demyelination and remyelination during EAE.In detail,M1 phenotype is an important cause of demyelination and detrimental to remyelination while M2 phenotype can promote remyelination and inhibit demyelination.In this review,we not only focus on advances in the direct mechanisms of microglial function on demyelination and remyelination in EAE model,also the indirect mechanisms by astrocytes.

[The application of pedicle orbital fat flap for the correction of lacrimal groove and palpebromalar groove deformity in the middle-aged and old people].

OBJECTIVE: To explore the effective techniques for correction of lacrimal groove and palpebromalar groove deformity in the middle-aged and old people. METHOD: The lacrimal groove and palpebromalar groove deformity was corrected by the techniques of transcutaneous orbital fat releasing and pedicle orbital fat flap filling. From 1996 to 2011, 426 patients, aged from 35 to 72 (average, 48), were treated by the techniques. Among them, 54 patients had underwent the surgical treatment before this operation. 362 patients were followed up for 3-24 months. RESULTS: Completely correction was achieved in 283 patients, obvious improvement in 79 patients. The result was not satisfied in 2 patients with severe deformity who had surgical treatment before. CONCLUSION: The lacrimal groove and palpebromalar groove deformity can be effectively corrected by transcutaneous orbital fat releasing and pedicle orbital fat flap in the middle-aged and old people.