Bazi Bushen ameliorates age-related energy metabolism dysregulation by targeting the IL-17/TNF inflammatory pathway associated with SASP.

BACKGROUND: Chronic inflammation and metabolic dysfunction are key features of systemic aging, closely associated with the development and progression of age-related metabolic diseases. Bazi Bushen (BZBS), a traditional Chinese medicine used to alleviate frailty, delays biological aging by modulating DNA methylation levels. However, the precise mechanism of its anti-aging effect remains unclear. In this study, we developed the Energy Expenditure Aging Index (EEAI) to estimate biological age. By integrating the EEAI with transcriptome analysis, we aimed to explore the impact of BZBS on age-related metabolic dysregulation and inflammation in naturally aging mice. METHODS: We conducted indirect calorimetry analysis on five groups of mice with different ages and utilized the data to construct EEAI. 12 -month-old C57BL/6 J mice were treated with BZBS or ß-Nicotinamide Mononucleotide (NMN) for 8 months. Micro-CT, Oil Red O staining, indirect calorimetry, RNA sequencing, bioinformatics analysis, and qRT-PCR were performed to investigate the regulatory effects of BZBS on energy metabolism, glycolipid metabolism, and inflammaging. RESULTS: The results revealed that BZBS treatment effectively reversed the age-related decline in energy expenditure and enhanced overall metabolism, as indicated by the aging index of energy expenditure derived from energy metabolism parameters across various ages. Subsequent investigations showed that BZBS reduced age-induced visceral fat accumulation and hepatic lipid droplet aggregation. Transcriptomic analysis of perirenal fat and liver indicated that BZBS effectively enhanced lipid metabolism pathways, such as the PPAR signaling pathway, fatty acid oxidation, and cholesterol metabolism, and improved glycolysis and mitochondrial respiration. Additionally, there was a significant improvement in inhibiting the inflammation-related arachidonic acid-linoleic acid metabolism pathway and restraining the IL-17 and TNF inflammatory pathways activated via senescence associated secretory phenotype (SASP). CONCLUSIONS: BZBS has the potential to alleviate inflammation in metabolic organs of naturally aged mice and maintain metabolic homeostasis. This study presents novel clinical therapeutic approaches for the prevention and treatment of age-related metabolic diseases.

Tripterygium wilfordii Hook.f. ameliorates paraquat-induced lung injury by reducing oxidative stress and ferroptosis via Nrf2/HO-1 pathway.

Paraquat (PQ) poisoning can induce acute lung injury and fibrosis and has an extremely high mortality rate. However, no effective treatments for PQ poisoning have been established. In this study, the potential efficacy of Tripterygium wilfordii Hook.f. (TwHF) in alleviating PQ-induced lung injury and fibrosis was investigated in a mouse model. Mice were randomly assigned to the control, PQ, PQ + TwHF1 (pretreatment before inducing poisoning), and PQ + TwHF2 (treatment after poisoning) groups. The mice in the PQ + TwHF1 group were pretreated with TwHF for 5 days before receiving one dose of PQ (120 mg/kg) and then received a daily oral gavage of the indicated dosages of TwHF until sacrifice. The mice in the PQ + TwHF2 group were treated with TwHF 2 h after PQ exposure until sacrifice. The pathological analysis and Fapi PET/CT showed that treatment with TwHF attenuated lung injury. And TwHF reduced pulmonary oxidative stress, as indicated by the reduction in, malondialdehyde (MDA), glutathione (GSH), and reactive oxygen species (ROS) levels, as well as by the increase in superoxide dismutase (SOD) levels. Accordingly, the Perls DAB staining showed increased iron concentrations and western blotting revealed a decreased GPX4 expression after PQ exposure, as well as the mitigation of the overexpression of Nrf2 and HO-1 induced by PQ. In conclusion, our study demonstrated the potential of TwHF as a treatment for PQ-induced lung injury and fibrosis. The protective mechanism of this medicinal herb may involve the regulation of ferroptosis.

Polyketides with anti-inflammatory activity isolated from Rhodiola tibetica endophytic fungus Penicillium sp. HJT-A-10.

Seven undescribed polyketide compounds (1-4, 9-11) and six known polyketide compounds (5-8,12, 13) were isolated from Rhodiola tibetica endophytic Penicillium sp. HJT-A-10. The structural of seven undescribed polyketides metabolites were established on the basis of spectroscopic methods. The results of anti-inflammatory activity showed that compounds 1-8，10-13 had significant inhibitory effects on LPS-induced NO production in RAW 264.7 cells.

Cordycepin enhances hyperthermia-induced apoptosis and cell cycle arrest by modulating the MAPK pathway in human lymphoma U937 cells.

BACKGROUND: Hyperthermia induces cancer cell death. However, the cytotoxic effect of hyperthermia is not sufficient. Cordycepin can also induce apoptosis in cancer cells and enhance the antitumoral activity of irradiation. To examine cordycepin-mediated enhancement of hyperthermia-induced apoptosis, this study investigated the combined effects and apoptotic mechanisms of hyperthermia and cordycepin on human leukemia U937 cells. METHODS: Cell viability and apoptosis were measured using MTT assays, Hoechst 33342 staining and Annexin V/PI double staining. The distribution of the cell cycle and sub-G1 phase, reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) were examined by flow cytometry. The expression of related proteins was analyzed by western blotting. RESULTS: Combined treatment with hyperthermia and cordycepin markedly augmented apoptosis by upregulating Bax and suppressing Bcl-2, Bid and activated caspase 3 and 8 expression, and apoptosis was decreased by Z-VAD-fmk (a pan caspase inhibitor). We also found that the MMP was significantly decreased and excessive ROS generation occurred. The combination treatment also induced arrest in the G2/M phase by downregulating cyclin dependent kinase 1 (CDK1) and cyclin B1 protein expression. Furthermore, it was observed that mitogen-activated protein kinase (MAPK) pathway including ERK, JNK and p38 signals was involved in the induction of apoptosis. The phosphorylated p38 and JNK were increased and ERK phosphorylation was decreased by the combined treatment. In addition, N-acetyl-L-cysteine (NAC) significantly protected the cells by restoring ROS levels and the activity of caspase-3, inactivating the MAPK pathway. CONCLUSION: Cordycepin significantly enhanced hyperthermia-induced apoptosis and G2/M phase arrest in U937 cells. The combined treatment enhanced apoptosis through the MAPK pathway and mitochondrial dysfunction, and these effects could be rescued by NAC. We report for the first time that cordycepin can be used as a hyperthermia sensitizer to treat leukemia.

Phytochemical, Chemotaxonomic and Bioinformatics Study on Laportea bulbifera (Urticaceae).

Phytochemical investigation of the aerial part of Laportea bulbifera (Siebold & Zucc.) Wedd. (L. bulbifera) showed the isolation of seventeen compounds, including five flavonoids (1-4 and 6), one terpenoid (5), five phenolic acids (7-11), one coumarin (12), two steroids (13-14), and three alkaloids (15-17). Structure elucidations of these compounds were performed on the basis of extensive NMR experiments and compared with the published data in the references. It is remarkable that compounds (3-5) were firstly isolated from the Urticaceae family, compounds (3-8, 11 and 15-17) were firstly obtained from genus Laportea. Furthermore, the result of the chemotaxonomic significance discussion showed that compounds (2-4) may can be served as compound fingerprints to distinguish between species of L. bulbifera and genus Urtica, and what' more, we proposed a bold conjecture that isoflavones can distinguish between species of L. bulbifera and genus Urtica. At the same time, the molecular docking method was used to evaluate the inhibitory effect of these compounds on human steroid 5α-reductase 2 (SRD5α2). The results showed that compounds (1-4 and 6) had better expected effects than the positive drug finasteride can by effectively binding to the active sites of SRD5α2. This study assisted in the future phytochemical and chemotaxonomic research on genus Laportea. Simultaneously, this research provided the theoretical evidence for the application of L. bulbifera in treating benign prostatic hyperplasia (BPH).

Supplementary research on K150del variant of activated protein C.

Activated protein C (APC) is an anticoagulant with potent cytoprotective and anti-inflammatory effects. K150del, a natural variant of APC, is associated with reduced anticoagulant activity. We performed a comprehensive study to analyze the functional alterations of the K150del mutant. Transcriptome analysis of HEK 293T cells treated with wild and mutant APC revealed differentially expressed genes enriched in inflammatory, apoptotic, and virus defense-related signaling pathways. Both wild and mutant APC displayed concentration-dependent cytoprotective effects. Low concentrations of K150del mutant resulted in decreased anti-inflammatory and anti-apoptotic activities, whereas its higher concentrations restored these effects. Expression of virus defense-related genes improved in mouse lung tissues after repeated administration of the APC variant. These results suggest that the APC K150del mutant could help clinicians to accurately predict disease risks and serve as a potential auxiliary therapeutic in viral infections, including 2019 coronavirus disease (COVID-19).

A practical and fast isolation of 12 cyclolinopeptides (linusorbs) from flaxseed oil via preparative HPLC with phenyl-hexyl column.

Linusorbs, known as cyclolinopeptides, are a group of cyclic hydrophobic peptides derived from flaxseed oil with various health benefits. However, the current research efforts on both the biological activities and antioxidant capacities of linusorbs are limited because of existing issues with their purification and characterization. A practical method based on preparative HPLC for isolating 12 linusorbs simultaneously was developed and factors such as the solvent selection, gradient elution program, flow rate, loaded mass, and loading concentration, were optimized. The optimum conditions were an initial acetonitrile (ACN) to water ratio of 40%, final ACN ratio of 80%, eluting time of 21 min, a flow rate of 16 mL/min, sample load of 12.5 mg, and concentration of 80 mg/mL (in methanol). The 12 linusorbs obtained were verified using off-line MS/MS, recording purities of above 95.5%. The method could serve as a practical and fast isolation method enabling further investigation of minor linusorbs.

[Clinical trials of acupuncture treatment of post-stroke shoulder pain].

OBJECTIVE: To observe the clinical therapeutic effect of "Tiaoshen Zhitong" (mental regulating and pain relieving) needling and its influence on serological indicators in the treatment of post-stroke shoulder pain, so as to provide new therapeutic thoughts and approach for post-stroke shoulder pain. METHODS: A total of 80 inpatients with post-stroke shoulder pain were randomly divided into a control group (routine needling, 39 cases) and an observation group ("Tiaoshen Zhitong" needling, 41 cases) according to the random number table. Patients of the two groups accepted basic medication treatment including anticoagulants, hypotensive drugs, hypoglycermic drugs, lipid-lowering drugs, etc. In addition, patients of the control group were also treated by routine acupuncture stimulation (uniform reinforcing-reducing stimulation) of Jianyu (LI15), Jianqian (EX-UE12), Jianhou (Extra), Jianliao (TE14), Waiguan (TE5) and Hegu (LI4) on the affected side, and those of the observation group also treated by "Tiaoshen Zhitong" needling of Ear-Shenmen (MA-TF1), bilateral Neiguan (PC6, lifting-thrusting-reducing method), Shuigou (GV26, lifting-thrusting-reducing method), and Jianyu (LI15), Jianliao(TE14), Jianzhen (SI9) and Yanglingquan (GB34, the latter 4 points were stimulated with uniform reinforcing-reducing method) on the affected side. The treatment was given once every day, 6 days a week for 4 weeks. The pain severity was assessed by using visual analogue scale (VAS), the upper limb function evaluated by using Fugl-Meyer assessment (FMA) scale, the shoulder-joint function evaluated by using Constant-Murley score (CMS) questionnaire, and the daily living ability assessed by using Barthel index (BI) scale. The enzyme linked immunosorbent assay (ELISA) was used to determine the contents of serum beta-endorphin (ß-EP), enkephalin (ENK) and dynorphin (Dyn). The clinical therapeutic effect was evaluated by using Nimodipine scale method. RESULTS: Of the 39 and 41 cases in the control and observation groups, 7(17.95%) and 12(29.27%) were basically cured, 12(30.77%) and 13(31.71%) experienced marked improvement, 8(20.51%) and 11(26.83%) were effective, 12(30.77%) and 5 (12.19%) failed, with the total effective rate being 69.23% and 87.80%, respectively. The effective rate of the observation group was significantly higher than that of the control group (P<0.05). After the treatment, the VAS score was obviously reduced (P<0.01), and the scores of FMA scale, CMS questionnaire and BI scale, and contents of serum ß-EP, ENK and Dyn were all increased obviously in the two groups compared with their own pre-treatment (P<0.01). The therapeutic effect of "Tiaoshen Zhitong" needling was significantly superior to that of the routine needling in lowering VAS, and in raising scores of FMA scale, CMS questionnaire and BI scale and in up-regulating serum ß-EP, ENK and Dyn levels (P<0.01). CONCLUSION: "Tiaoshen Zhitong" needling is effective in reducing post-stroke shoulder pain and improving the motor function of the upper limb and shoulder-joint as well as the quality of daily life in stroke patients with shoulder pain. Its analgesic effect is probably related to the increase of the levels of serum ß-EP, ENK and Dyn.

The ameliorative effect of bloodletting puncture at hand twelve Jing-well points on cerebral edema induced by permanent middle cerebral ischemia via protecting the tight junctions of the blood-brain barrier.

BACKGROUND: Cerebral edema, erupting simultaneously with severe ischemic stroke, might lead to increased intracranial pressure, cerebral herniation, and ultimately death. Studies conducted previously by our team have demonstrated the fact that bloodletting puncture at hand twelve Jing-well points (HTWP) could alleviate cerebral edema, which mainly results from the disruption of blood-brain barrier (BBB). The study, therefore, was first designed to demonstrate whether BBB-protection serves an important role in the edema-relief effect of HTWP bloodletting, based on which to research the molecular mechanism underlying. METHODS: The rats were made into model suffering from permanent middle cerebral artery occlusion (pMCAO) and then bloodletting puncture were treated at HTWP once a day. Wet and dry weight method was adopted to evaluate the degree of brain edema, evans blue extravasation and electron microscopy were used to evaluate the integrity of the BBB, and RT-qPCR was carried out to analyze the expression level of occludin, claudin-5, ICAM-1, and VEGF. RESULTS: Results revealed that bloodletting puncture treatment could reduce water content of brain and the permeability of BBB caused by ischemic stroke. In bloodletting puncture group, ameliorated tight junctions could be observed under electron microscopy. It was demonstrated in further study that, in bloodletting group, compared with pMCAO one, the expression levels of occludin and claudin-5 were up-regulated, while ICAM-1 and VEGF were down-regulated. CONCLUSIONS: In conclusion, bloodletting puncture at HTWP might play a significant role in protecting the tight junctions of BBB, thus alleviating cerebral edema induced by ischemic stroke. Therefore, the therapy of bloodletting puncture at HTWP may be a promising strategy for acute ischemic stroke in the future.

Ethyl acetate extract of Peperomia tetraphylla induces cytotoxicity, cell cycle arrest, and apoptosis in lymphoma U937 cells.

The current study evaluated the cytotoxicity and the mechanism of apoptotic induction by Peperomia tetraphylla in U937 lymphoma cells. The results showed that P. tetraphylla ethyl acetate extract (EAEPT) inhibited the cell growth in U937 cells by MTT assay. After the U937 cells were treated with EAEPT, the cells exhibited marked morphological features of apoptosis (Hoechst 33342 staining) and the number of apoptotic cell (Annexin V-FITC/PI staining) increased. The treatment of EAEPT could induce loss of mitochondrial membrane potential (MMP) and increase the ROS level. Moreover, EAEPT treatment resulted in the accumulation of cells at S phase. We found that EAEPT could induce the cleavage of the caspase 3, caspase 8, caspase 9 and Bid. And the treatment of EAEPT could increase expression of Bax and down-regulate the expression of CCNB1, CCND1 and CDK1. The sub-fraction of EAEPT, namely EASub1 demonstrated the highest cytotoxicity activity on U937 cells. It was confirmed that EAEPT could inhibit the growth of U937 cells by blocking the cell cycle and prompted apoptosis via the ROS-medicated mitochondria pathway in vitro.

New steroidal glycosides from Tribulus terrestris L.

Two new steroidal glycosides were isolated from Tribulus terrestris L. Their structures were elucidated as 26-O-ß-D-glucopyranosyl-5α-furostan-12-one-20(22)-ene-3ß,23,26-triol-3-O-ß-D-xylopyranosyl-(1 → 2)-[ß-D-xylopyranosyl-(1 → 3)]-ß-D-glucopyranosyl-(1 → 4)-[α-L-rhamnopyranosyl-(1 → 2)]-ß-D-galactopyranoside (1) and 26-O-ß-D-glucopyranosyl-5α-furostan-20(22)-ene-3ß,23,26-triol-3-O-ß-D-xylopyranosyl-(1 → 2)-[ß-D-xylopyranosyl-(1 → 3)]-ß-D-glucopyranosyl-(1 → 4)-[α-L-rhamnopyranosyl-(1 → 2)]-ß-D-galactopyranoside (2) by spectroscopic methods including 1D and 2D NMR experiments.

Chemical constituents from the fruits of Trichosanthes kirilowii.

Two new compounds, 1-deoxy-1-[2'-oxo-1'-pyrrolidinyl]-2-n-butyl-α-fructofuranoside and isoarvenin III, were isolated from the fruits of Trichosanthes kirilowii Maxim, and their structures were established on the basis of spectroscopic methods.

Two new steroidal glycosides from Cynanchum amplexicaule.

Two new C21 steroidal glycosides were isolated from the EtOAc fraction of Cynanchum amplexicaule. Their structures were elucidated as amplexicogenin D-3-O-ß-D-cymaropyranoside and stauntogenin -3-O-α-L-oleandropyranosyl-(1 → 4)-O-ß-D-digitoxopyranosyl-(1 → 4)-O-ß-D-oleandropyranoside by spectroscopic analysis including 1D and 2D NMR experiments.

Aromatic compounds from endophytic fungus Colletotrichum sp. L10 of Cephalotaxus hainanensis Li.

Six aromatic compounds were isolated from the endophytic fungus Colletotrichum sp. of Cephalotaxus hainanensis Li [Cephalotaxus mannii Hook. f.]. Their structures were determined on the basis of chemical and spectroscopic methods. The compound 2 was the enantiomer of compound 1. The compound 4 was the epimeride of the compound 3. The compounds 1, 2, 4 and 5 were evaluated for cytostatic activity against HL-60 and PC-3 cells.

Two new compounds from the fruits of Trichosanthes kirilowii Maxim.

Two new compounds 1,3-O-[5-(hydroxymethyl)-furan-2-yl]methenyl-2-n-butyl-α-fructofuranoside and n-butyl-3,4-dihydroxyl-5-hydroxymethyl-4-O-[5-(hydroxymethyl)-furan-2-yl]-tetrahydrofuran-2-carboxylate were isolated from the fruits of Trichosanthes kirilowii Maxim, and their structures were established on the basis of spectroscopic methods.

Three new compounds from Dictamnus dasycarpus.

Phytochemical investigation of the ethyl acetate extracts from root barks of Dictamnus dasycarpus led to the isolation of three new compounds, named as dasycarpusenester A (1), dasycarpusester B (2), dasycarpusacid (3). Their structures were elucidated as (2S)-4-(2,2-dimethyl-5-oxotetrahydrofuran-3-yl)-2-hydroxypent-3-enoic acid methyl ester (1), (2R)-4-(2,2-dimethyl-5-oxotetrahydrofuran-3-yl)-2-hydroxypent-3-anoic acid methyl ester (2), and (2S)-4-(2,2-dimethyl-5-oxotetrahydrofuran-3-yl)-2-hydroxypent-3-anoic acid (3), respectively, on the basis of modern spectroscopic methods and chemical analysis.

[Study on the mechanisms of cupping therapy].

Computerized literature searches are performed for articles of the mechanism of cupping therapy in the past 10 years. The mechanism of action of local negative pressure applied to human body is studied in terms of specific changes in local tissue structure, stretch to the nerve and muscle, increasing blood circulation and causing autohemolysis. This paper aims to explain the mechanism of cupping therapy according to modern science.

Two new compounds from an endophytic fungus Aspergillus sp. HS-05.

Two new compounds, N-[4'-hydroxy-(E)-cinnamoyl]-l-tyrosine methyl ester (1) and methyl 4-methoxy-3-(3'-hydroxy-2'-methyl)propionyloxy-benzoate (2), were isolated from EtOAc extract of the fermentation broth of the endophytic fungus Aspergillus sp. HS-05. Their structures were elucidated by NMR, IR, UV, MS, and CD methods. Compounds 1 and 2 showed no anticancer activities against HL-60 cell lines (both of IC(50)>100 µM) in vitro.

[Medicinal cupping: clinical application].

The commonly used tools and key points of manipulations of medicinal cupping therapy were introduced systematically in this article in order to study the concerning apparatuses, therapeutic methods, detailed manipulations, indications, application locations and medicine selection. Thus, an overall comprehension of the clinical application of medicinal cupping can be acquired, which may provide train of thought and methodology for its clinical application.

Two new steroidal saponins from Tribulus terrestris L.

Two new steroidal saponins were isolated from the fruits of Tribulus terrestris L. Their structures were elucidated by spectroscopic and chemical analysis as (23S,24R,25R)-5alpha-spirostane-3beta,23,24-triol-3-O-{alpha-L-rhamnopyranosyl-(1 --> 2)-[beta-D-glucopyranosyl-(1 --> 4)]-beta-D-galactopyranoside} (1) and (23S,24R,25S)-5alpha-spirostane-3beta,23,24-triol-3-O-{alpha-L-rhamnopyranosyl-(1 --> 2)-[beta-D-glucopyranosyl-(1 --> 4)]-beta-D-galactopyranoside} (2).