Structural insights into substrate recognition and translocation of human peroxisomal ABC transporter ALDP.

Dysfunctions of ATP-binding cassette, subfamily D, member 1 (ABCD1) cause X-linked adrenoleukodystrophy, a rare neurodegenerative disease that affects all human tissues. Residing in the peroxisome membrane, ABCD1 plays a role in the translocation of very long-chain fatty acids for their ß-oxidation. Here, the six cryo-electron microscopy structures of ABCD1 in four distinct conformational states were presented. In the transporter dimer, two transmembrane domains form the substrate translocation pathway, and two nucleotide-binding domains form the ATP-binding site that binds and hydrolyzes ATP. The ABCD1 structures provide a starting point for elucidating the substrate recognition and translocation mechanism of ABCD1. Each of the four inward-facing structures of ABCD1 has a vestibule that opens to the cytosol with variable sizes. Hexacosanoic acid (C26:0)-CoA substrate binds to the transmembrane domains (TMDs) and stimulates the ATPase activity of the nucleotide-binding domains (NBDs). W339 from the transmembrane helix 5 (TM5) is essential for binding substrate and stimulating ATP hydrolysis by substrate. ABCD1 has a unique C-terminal coiled-coil domain that negatively modulates the ATPase activity of the NBDs. Furthermore, the structure of ABCD1 in the outward-facing state indicates that ATP molecules pull the two NBDs together and open the TMDs to the peroxisomal lumen for substrate release. The five structures provide a view of the substrate transport cycle and mechanistic implication for disease-causing mutations.

Activities of daily living and its influencing factors for older people with type 2 diabetes mellitus in urban communities of Fuzhou, China.

Background: Type 2 diabetes mellitus (T2DM) is an independent risk factor for functional limitations among the older population. The predicted increase in T2DM cases combined with the ongoing rapidly aging population may further burden the already overloaded healthcare system and aggravate the loss of economic self-sufficiency. This study aimed to investigate the activities of daily living (ADL) and its influencing factors on older people with T2DM, and to provide implications for the development and improvement of community nursing services in the context rapidly aging population in China. Methods: From March 2019 to June 2020, we conducted a cross-sectional questionnaire survey among older T2DM patients in Fuzhou, using a multi-stage cluster sampling approach. Functional status was measured by the Lawton ADL scale. Stata "nptrend" test was used to examine the trend of ordinal variables on ADL. Non-conditional logistic regression was used to identify factors affecting ADL limitations. Results: A total of 2016 questionnaires were received, with a response rate of 96%. 12.4% of participants suffered from varying degrees of functional impairment. ADL limitations increased with age. More comorbidities were associated with a greater risk of developing functional limitations in ADLs. the following sub-groups were more likely to suffer from ADL impairment: those aged 70 and over years (OR = 1.99, 95%CI 1.77-2.56), living in an aged care house or with spouse/children (OR = 2.31, 95%CI 1.25-4.26), low monthly income (OR = 1.49, 95%CI 1.28-1.64), without health insurance (OR = 1.82, 95%CI 1.40-2.40), tight family expenses (OR = 1.95, 95%CI 1.42-2.69), having stroke (OR = 6.70, 95%CI 2.22-20.23) or malignant tumor (OR = 4.45, 95%CI 1.27-15.53), irregular eating habit (OR = 2.55, 95%CI 2.23-2.92), smoking (OR = 1.40, 95%CI 1.22-1.60), sedentary lifestyle (OR = 2.04, 95%CI 1.46-2.85), lack of physical exercise (OR = 1.35, 95%CI 1.19-1.53), sleeping difficulty (OR = 1.25, 95%CI 1.10-1.42), and lack of family support (OR = 1.19, 95%CI 1.10-1.29). Conclusion: Older adults (≥70 years) with T2DM had a high prevalence of functional limitations across a range of daily living tasks, which not only affect individual life of quality but also present a huge burden on the family, health services system, and the whole society. Identified factors associated with ADL limitations may provide useful information for targeted nursing practice and health promotion.

Seed oil of Brucea javanica induces apoptosis through the PI3K/Akt signaling pathway in acute lymphocytic leukemia Jurkat cells.

Brucea javanica oil emulsion (BJOE) has been used to treat tumor in China for more than 40 years. However, its components and effectiveness in the treatment of acute lymphocytic leukemia (ALL) and its mechanism of anti-cancer activity remain unknown. In the current study, high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) was used to analyze the components of BJOE. Then, the anti-leukemia effects of BJOE were examined both in vitro and in vivo using ALL Jurkat cells and the p388 mouse leukemia transplant model, respectively. The primary ALL leukemia cells were also used to confirm the anti-leukemia effects of BJOE. The apoptotic-related results indicated that BJOE induced apoptosis in Jurkat cells and were suggestive of intrinsic apoptotic induction. Moreover, BJOE inhibited Akt (protein kinase B) activation and upregulated its downstream targets p53 and FoxO1 (forkhead box gene, group O-1) to initiate apoptosis. The activation of GSK3ß was also involved. Our findings demonstrate that BJOE has anti-leukemia effects on ALL cells and can induce apoptosis in Jurkat cells through the phosphoinositide3-kinase (PI3K) /Akt signaling pathway.

pNNS-Conjugated Chitosan Mediated IGF-1 and miR-140 Overexpression in Articular Chondrocytes Improves Cartilage Repair.

The aim of the present study was to investigate the effects of phosphorylatable nucleus localization signal linked nucleic kinase substrate short peptide (pNNS)-conjugated chitosan (pNNS-CS) mediated miR-140 and IGF-1 in both rabbit chondrocytes and cartilage defects model. pNNS-CS was combined with pBudCE4.1-IGF-1, pBudCE4.1-miR-140, and negative control pBudCE4.1 to form pDNA/pNNS-CS complexes. Then these complexes were transfected into chondrocytes or injected intra-articularly into the knee joints. High levels of IGF-1 and miR-140 expression were detected both in vitro and in vivo. Compared with pBudCE4.1 group, in vitro, the transgenic groups significantly promoted chondrocyte proliferation, increased glycosaminoglycan (GAG) synthesis, and ACAN, COL2A1, and TIMP-1 levels, and reduced the levels of nitric oxide (NO), MMP-13, and ADAMTS-5. In vivo, the exogenous genes enhanced COL2A1, ACAN, and TIMP-1 expression in cartilage and reduced cartilage Mankin score and the contents of NO, IL-1ß, TNF-α, and GAG contents in synovial fluid of rabbits, MMP-13, ADAMTS-5, COL1A2, and COL10A1 levels in cartilage. Double gene combination showed better results than single gene. This study indicate that pNNS-CS is a better gene delivery vehicle in gene therapy for cartilage defects and that miR-140 combination IGF-1 transfection has better biologic effects on cartilage defects.

Targeting ALDH2 with disulfiram/copper reverses the resistance of cancer cells to microtubule inhibitors.

Disulfiram (DSF) in combination with copper (Cu) has been reported to override drug resistance in cancer cells, and DSF combined with chemotherapy based on the microtubule inhibitor vinorelbine appears to prolong survival in non-small cell lung cancer patients. Here, we investigated the mechanisms underlying these findings. DSF/Cu reversed the microtubule inhibitor resistance in A549/Taxol and KB/VCR cells in vitro, and had anti-tumor effects in A549/Taxol and KB/VCR xenograft mice. DSF/Cu and DSF reduced the cancer stem cell (CSC) characteristics of drug-resistant A549/Taxol and KB/VCR cells, including sphere formation, colony generation and migration, and DSF/Cu was more effective than DSF alone. DSF/Cu also decreased the aldehyde dehydrogenase (ALDH) activity and the expression of P-gp and stem cell transcription factors in A549/Taxol and KB/VCR cells. Knockdown of ALDH2 attenuated the CSC characteristics of resistant cancer cells and enhanced their sensitivity to Taxol or VCR. Importantly, DSF/Cu treatment inhibited the expression of ALDH2 in vitro and in vivo. Our findings suggest that DSF/Cu reverses microtubule inhibitor resistance in cancer cells by suppressing ALDH2 expression, and Cu improves the activity of DSF.

[Clinical observation on treatment of 43 women with polycystic ovary syndrome based on syndrome differentiation].

OBJECTIVE: To investigate the therapeutic effect of treatment based on syndrome differentiation and its influence on the serum levels of testosterone and insulin in women with polycystic ovary syndrome (PCOS). METHODS: Forty-three women, diagnosed with PCOS as well as syndrome of kidney yin deficiency or syndrome of spleen qi deficiency, were enrolled in the study. Twenty-five PCOS women with syndrome of kidney yin deficiency were treated with traditional Chinese herbs for nourishing yin to reduce fire (nourishing yin group), and 18 PCOS women with syndrome of spleen qi deficiency were treated with herbs for invigorating spleen and replenishing qi (replenishing qi group). Body mass index (BMI), waist-to-hip ratio (WHR), waist circumference (WC), and the serum levels of insulin and testosterone before and after treatment were detected. RESULTS: Among the 43 cases, 20 cases had high serum testosterone level, in which 13 cases with syndrome of kidney yin deficiency, 7 cases with syndrome of spleen qi deficiency, but the difference had no statistic significance; 17 cases had high serum insulin level, in which 11 cases with syndrome of spleen qi deficiency, 6 cases with syndrome of kidney yin deficiency, the difference had statistic significance (P<0.05). In nourishing yin group, the serum levels of insulin and testosterone declined after treatment (P<0.05), and BMI, WHR and WC showed no significant changes. In replenishing qi group, the serum level of insulin declined after treatment (P<0.01), and BMI, WHR, WC and the serum level of testosterone showed no significant changes. CONCLUSION: Traditional Chinese herbs for nourishing yin to reduce fire can significantly reduce the serum levels of testosterone and insulin in PCOS women with syndrome of kidney yin deficiency, and herbs for invigorating spleen and replenishing qi can significantly reduce the serum level of insulin in PCOS women with syndrome of spleen qi deficiency.