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Article ; Online: High-Fidelity Assay Based on Turn-Off Fluorescence to Detect the Perturbations of Cellular Proteostasis.

Hoelzel, Conner / Bai, Yulong / Wang, Mengdie / Liu, Yu / Zhang, Xin

ACS bio & med chem Au

2024 Volume 4, Issue 2, Page(s) 111–118

Abstract: The persistence of neurodegenerative diseases has necessitated the development of new strategies to monitor protein homeostasis (proteostasis). Previous efforts in our laboratory have focused on the development of fluorogenic strategies to observe the ... ...

Abstract	The persistence of neurodegenerative diseases has necessitated the development of new strategies to monitor protein homeostasis (proteostasis). Previous efforts in our laboratory have focused on the development of fluorogenic strategies to observe the onset and progression of proteostatic stress. These works utilized solvatochromic and viscosity sensitive fluorophores to sense protein folded states, enabling stressor screening with an increase in the emission intensity upon aggregation. In this work, we present a novel, high-fidelity assay to detect perturbations of cellular proteostasis, where the fluorescence intensity decreases with the onset of proteostatic stress. Utilizing a fluorogenic, hydroxymethyl silicon-rhodamine probe to differentiate between protein folded states, we establish the validity of this technology in living cells by demonstrating a two-fold difference in fluorescence intensity between unstressed and stressed conditions.
Language	English
Publishing date	2024-01-19
Publishing country	United States
Document type	Journal Article
ISSN	2694-2437
ISSN (online)	2694-2437
DOI	10.1021/acsbiomedchemau.3c00012
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Article ; Online: Effect of the Competing Ligand on Ni-NTA/Histag Strength Revealed by Click Chemistry-Based Force Spectroscopy.

Nie, Jingyuan / Wang, Ziyi / Wang, Mengdie / Zheng, Peng

The journal of physical chemistry letters

2023 Volume 14, Issue 23, Page(s) 5426–5431

Abstract: The specific interaction between Ni-nitrilotriacetic acid and the six-histidine tag may be one of the most important coordination bonds utilized in biological research because of its wide application for recombinant protein purification. The complex ... ...

Abstract	The specific interaction between Ni-nitrilotriacetic acid and the six-histidine tag may be one of the most important coordination bonds utilized in biological research because of its wide application for recombinant protein purification. The complex stability is critical for target protein binding. Thus, measurement of the mechanical stability of the system was attempted soon after the invention of atomic force microscopy-based single-molecule force spectroscopy (AFM-SMFS) two decades ago. Moreover, the competing ligand imidazole and protons are the two critical factors for target protein elution. However, the mechanochemistry between the system and the imidazole/proton has not been determined. Here, an AFM-SMFS system using strain-promoted alkyne-azide cycloaddition and Cu-free click chemistry was used to characterize the system. Consequently, the destabilizing effect of the imidazole and proton on the interaction was revealed quantitatively, leading to a 3-fold increase in the bond dissociation rate.
Language	English
Publishing date	2023-06-07
Publishing country	United States
Document type	Journal Article
ISSN	1948-7185
ISSN (online)	1948-7185
DOI	10.1021/acs.jpclett.3c00813
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Article ; Online: Quantification of carboxylate-bridged di-zinc site stability in protein due ferri by single-molecule force spectroscopy.

Wang, Zhiyi / Wang, Mengdie / Zhao, Zhongxin / Zheng, Peng

Protein science : a publication of the Protein Society

2023 Volume 32, Issue 3, Page(s) e4583

Abstract: Carboxylate-bridged diiron proteins belong to a protein family involved in different physiological processes. These proteins share the conservative EXXH motif, which provides the carboxylate bridge and is critical for metal binding. Here, we choose de ... ...

Abstract	Carboxylate-bridged diiron proteins belong to a protein family involved in different physiological processes. These proteins share the conservative EXXH motif, which provides the carboxylate bridge and is critical for metal binding. Here, we choose de novo-designed single-chain due ferri protein (DFsc), a four-helical protein with two EXXH motifs as a model protein, to study the stability of the carboxylate-bridged di-metal binding site. The mechanical and kinetic properties of the di-Zn site in DFsc were obtained by atomic force microscopy-based single-molecule force spectroscopy. Zn-DFsc showed a considerable rupture force of ~200 pN, while the apo-protein is mechanically labile. In addition, multiple rupture pathways were observed with different probabilities, indicating the importance of the EXXH-based carboxylate-bridged metal site. These results demonstrate carboxylate-bridged di-metal site is mechanically stable and improve our understanding of this important type of metalloprotein.
MeSH term(s)	Zinc/chemistry ; Metalloproteins/chemistry ; Metals ; Spectrum Analysis
Chemical Substances	Zinc (J41CSQ7QDS) ; Metalloproteins ; Metals
Language	English
Publishing date	2023-01-30
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1106283-6
ISSN	1469-896X ; 0961-8368
ISSN (online)	1469-896X
ISSN	0961-8368
DOI	10.1002/pro.4583
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Article ; Online: Suberoylanilide hydroxamic acid attenuates cognitive impairment in offspring caused by maternal surgery during mid-pregnancy.

Feng, Yunlin / Qin, Jia / Lu, Yanfei / Wang, Mengdie / Wang, Shengqiang / Luo, Foquan

PloS one

2024 Volume 19, Issue 3, Page(s) e0295096

Abstract: Some pregnant women have to experience non-obstetric surgery during pregnancy under general anesthesia. Our previous studies showed that maternal exposure to sevoflurane, isoflurane, propofol, and ketamine causes cognitive deficits in offspring. Histone ... ...

Abstract	Some pregnant women have to experience non-obstetric surgery during pregnancy under general anesthesia. Our previous studies showed that maternal exposure to sevoflurane, isoflurane, propofol, and ketamine causes cognitive deficits in offspring. Histone acetylation has been implicated in synaptic plasticity. Propofol is commonly used in non-obstetric procedures on pregnant women. Previous studies in our laboratory showed that maternal propofol exposure in pregnancy impairs learning and memory in offspring by disturbing histone acetylation. The present study aims to investigate whether HDAC inhibitor suberoylanilide hydroxamic acid (SAHA) could attenuate learning and memory deficits in offspring caused by maternal surgery under propofol anesthesia during mid-pregnancy. Maternal rats were exposed to propofol or underwent abdominal surgery under propofol anesthesia during middle pregnancy. The learning and memory abilities of the offspring rats were assessed using the Morris water maze (MWM) test. The protein levels of histone deacetylase 2 (HDAC2), phosphorylated cAMP response-element binding (p-CREB), brain-derived neurotrophic factor (BDNF), and phosphorylated tyrosine kinase B (p-TrkB) in the hippocampus of the offspring rats were evaluated by immunofluorescence staining and western blot. Hippocampal neuroapoptosis was detected by TUNEL staining. Our results showed that maternal propofol exposure during middle pregnancy impaired the water-maze learning and memory of the offspring rats, increased the protein level of HDAC2 and reduced the protein levels of p-CREB, BDNF and p-TrkB in the hippocampus of the offspring, and such effects were exacerbated by surgery. SAHA alleviated the cognitive dysfunction and rescued the changes in the protein levels of p-CREB, BDNF and p-TrkB induced by maternal propofol exposure alone or maternal propofol exposure plus surgery. Therefore, SAHA could be a potential and promising agent for treating the learning and memory deficits in offspring caused by maternal nonobstetric surgery under propofol anesthesia.
MeSH term(s)	Humans ; Pregnancy ; Rats ; Animals ; Female ; Propofol/adverse effects ; Vorinostat/pharmacology ; Brain-Derived Neurotrophic Factor/metabolism ; Histones/metabolism ; Maze Learning ; Cognitive Dysfunction/chemically induced ; Cognitive Dysfunction/metabolism ; Hippocampus/metabolism ; Memory Disorders/chemically induced ; Memory Disorders/metabolism ; Anesthesia, General
Chemical Substances	Propofol (YI7VU623SF) ; Vorinostat (58IFB293JI) ; Brain-Derived Neurotrophic Factor ; Histones
Language	English
Publishing date	2024-03-29
Publishing country	United States
Document type	Journal Article
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0295096
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Article ; Online: Characterization of Transthyretin Mutation G47V Associated with Hereditary Cardiac Amyloidosis.

He, Xiaopeng / Wang, Mengdie / Sun, Jialu / Yu, Zhengyang / Hu, Xinyang / Liu, Yu / Lin, Xiaoping

Cardiology

2024

Abstract: Introduction: Amyloidosis caused by TTR mutations (ATTRv) is a rare inherited and autosomal dominant disease. More than 150 mutants of TTR have been reported, whereas some of them remain to be investigated.: Methods: A 52-year-old male presented with ...

Abstract	Introduction: Amyloidosis caused by TTR mutations (ATTRv) is a rare inherited and autosomal dominant disease. More than 150 mutants of TTR have been reported, whereas some of them remain to be investigated. Methods: A 52-year-old male presented with heart failure and clinically diagnosed ATTR cardiac amyloidosis (ATTR-CA) was recruited. Whole exome sequencing (WES) was performed. Biochemical and biophysical experiments characterized protein stability using urea-mediated tryptophan fluorescence. Drug response was analyzed by fibril formation assay. Finally, tetramer TTR concentration in patient' serum sample was measured by ultra-performance liquid chromatography (UPLC). Results: For the proband, whole exome sequencing revealed a mutation (c.200G>T; p.Gly67Val and referred to as G47V) in TTR gene. Biochemical and biophysical kinetics study showed that the thermodynamic stability of G47V-TTR (Cm = 2.4 M) was significantly lower than that of WT-TTR (Cm = 3.4 M) and comparable to that of L55P-TTR (Cm = 2.3 M), an early age-of-onset mutation. G47V:WT-TTR heterozygous tetramers kinetic stability (t1/2 = 1.4 h) was further compromised compared to that of the homozygous G47V-TTR (t1/2 = 3.1 h). Among three small molecule stabilizers, AG10 exhibited the best inhibition of the fibrillation of G47V-TTR homozygous protein. Using a UPLC assay, nearly 40% of TTR in this patient was calculated to be non-tetrameric. Conclusion: In this work, we reported a patient presented early onset of clinically typical ATTR-CM due to G47V-TTR mutation. Our work not only for the first time characterized the biochemical properties of G47V-TTR mutation, but also provided hints for the pathogenicity of this mutation.
Language	English
Publishing date	2024-03-04
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	80092-2
ISSN	1421-9751 ; 0008-6312
ISSN (online)	1421-9751
ISSN	0008-6312
DOI	10.1159/000538081
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Article ; Online: Transaxillary Single-Port Endoscopic Nipple-Sparing Mastectomy with Immediate Implant-based Breast Reconstruction in Breast Cancer Patients Receiving Neoadjuvant Chemotherapy or Not: A Comparative Study with Analysis of Surgical Complications and Patient-Reported Outcomes.

Zhao, Jingjing / Chen, Zujin / Wang, Mengdie / Hai, Linyue / Xiao, Chunhua

Aesthetic plastic surgery

2023 Volume 47, Issue 6, Page(s) 2304–2321

Abstract: Background: In most cases, transaxillary single-port endoscopic nipple-sparing mastectomy with immediate implant-based breast reconstruction (E-NSM-IIBR) is conducted in patients with early-stage breast cancer, ensuring surgical safety while achieving ... ...

Abstract	Background: In most cases, transaxillary single-port endoscopic nipple-sparing mastectomy with immediate implant-based breast reconstruction (E-NSM-IIBR) is conducted in patients with early-stage breast cancer, ensuring surgical safety while achieving improved breast aesthetics. However, whether E-NSM-IIBR is appropriate in patients undergoing neoadjuvant chemotherapy (NAC) is still unclear. The aim of this study was to report the surgical safety and patient-reported outcomes (PROs) of breast cancer patients who underwent E-NSM-IIBR with NAC in comparison to those who did not receive NAC. Methods: A retrospective cohort study was conducted on patients who underwent E-NSM-IIBR with or without NAC at a single center between January 2021 and July 2022. Patient demographics, postoperative complications, and PROs evaluated using the BREAST-Q version 2.0 questionnaire were compared between the two groups. Factors associated with PROs at 9 months after surgery were assessed with linear regression analysis. Results: A total of 92 patients who underwent E-NSM-IIBR were included in the study, with 27 patients receiving NAC and 65 patients not receiving NAC. There was no significant difference in the incidence of postoperative complications between the two groups. The BREAST-Q version 2.0 questionnaire was completed by 24 out of 27 patients (88.9%) in the NAC group and 59 out of 65 patients (90.8%) in the non-NAC group at 9 months after surgery. The patient-reported outcomes in various domains of the BREAST-Q did not show a significant difference between the two cohorts. The results of the multiple linear regression analysis indicated that in the both groups age (β = - 0.985, 95% CI - 1.598 to - 0.371, p = 0.003 in the NAC group; β = - 0.510, - 1.011 to - 0.009, p = 0.046 in the non-NAC group) and rippling (β = - 21.862, - 36.768 to - 6.955, p = 0.006 in the NAC group; β = - 7.787, - 15.151 to - 0.423, p = 0.039 in the non-NAC group) significantly impacted the patients' satisfaction with breasts, and PMRT was negatively associated with patients' physical well-being of chest (β = - 13.813, - 26.962 to - 0.664, p = 0.040 in the NAC group; β = - 18.574, - 30.661 to - 6.487, p = 0.003 in the non-NAC group). Our findings revealed that patients with larger implant volumes had higher scores in psychosocial well-being (β = 0.082, 0.001 to 0.162, p = 0.047), whereas implant displacement (β = - 14.937, - 28.175 to - 1.700, p=0.028) had a negative impact on patients' psychological well-being in the non-NAC group. However, our results did not demonstrate any significant influencing factors on patients' psychosocial well-being within the NAC group. Conclusion: Our preliminary experiences confirm that E-NSM-IIBR is a safe option for selected patients even after NAC, with favorable patient-reported outcomes comparable with those in the primary surgery setting. The postoperative long-term outcomes of patients who undergo radiation therapy after NAC merit further investigation in the future. Level of evidence iii: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .
MeSH term(s)	Humans ; Female ; Breast Neoplasms/drug therapy ; Breast Neoplasms/surgery ; Mastectomy/methods ; Retrospective Studies ; Neoadjuvant Therapy ; Nipples/surgery ; Mammaplasty/adverse effects ; Mammaplasty/methods ; Postoperative Complications/epidemiology ; Postoperative Complications/surgery ; Patient Reported Outcome Measures
Language	English
Publishing date	2023-09-12
Publishing country	United States
Document type	Journal Article
ZDB-ID	532791-x
ISSN	1432-5241 ; 0364-216X
ISSN (online)	1432-5241
ISSN	0364-216X
DOI	10.1007/s00266-023-03644-0
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Article: Comparative Pharmacokinetic Study of Three Major Bioactive Components in the Normal and Different Courses of Liver-Stagnation and Spleen-Deficiency Syndrome Depressive Rats after Intragastric Administration of Zhi-Zi-Hou-Po Decoction.

Luo, Kaiwen / Wang, Mengdie / Pan, Xiangxiang / Xing, Yadong

Journal of analytical methods in chemistry

2022 Volume 2022, Page(s) 8657616

Abstract: To reveal the pharmacokinetic process of narirutin, naringin, and honokiol in normal and different courses of liver-stagnation and spleen-deficiency syndrome depressive rats after intragastric administration of Zhi-Zi-Hou-Po decoction (ZZHPD), a rapid ... ...

Abstract	To reveal the pharmacokinetic process of narirutin, naringin, and honokiol in normal and different courses of liver-stagnation and spleen-deficiency syndrome depressive rats after intragastric administration of Zhi-Zi-Hou-Po decoction (ZZHPD), a rapid ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) method was proposed in this study. Chronic unpredictable mild stress (CUMS) rat was employed as the depression model. Acetonitrile solution containing 0.1% acetic acid and methyl alcohol-water (50 : 50,
Language	English
Publishing date	2022-03-24
Publishing country	Egypt
Document type	Journal Article
ZDB-ID	2654178-6
ISSN	2090-8873 ; 2090-8865
ISSN (online)	2090-8873
ISSN	2090-8865
DOI	10.1155/2022/8657616
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Article ; Online: PD-LI promotes rear retraction during persistent cell migration by altering integrin β4 dynamics.

Wang, Mengdie / Xiong, Choua / Mercurio, Arthur M

The Journal of cell biology

2022 Volume 221, Issue 5

Abstract: Although the immune checkpoint function of PD-L1 has dominated its study, we report that PD-L1 has an unanticipated intrinsic function in promoting the dynamics of persistent cell migration. PD-L1 concentrates at the rear of migrating carcinoma cells ... ...

Abstract	Although the immune checkpoint function of PD-L1 has dominated its study, we report that PD-L1 has an unanticipated intrinsic function in promoting the dynamics of persistent cell migration. PD-L1 concentrates at the rear of migrating carcinoma cells where it facilitates retraction, resulting in the formation of PD-L1-containing retraction fibers and migrasomes. PD-L1 promotes retraction by interacting with and localizing the β4 integrin to the rear enabling this integrin to stimulate contractility. This mechanism involves the ability of PD-L1 to maintain cell polarity and lower membrane tension at the cell rear compared with the leading edge that promotes the localized interaction of PD-L1 and the β4 integrin. This interaction enables the β4 integrin to engage the actin cytoskeleton and promote RhoA-mediated contractility. The implications of these findings with respect to cell-autonomous functions of PD-L1 and cancer biology are significant.
MeSH term(s)	Actin Cytoskeleton/metabolism ; B7-H1 Antigen/metabolism ; Cell Movement ; Cell Polarity ; Integrin beta4/metabolism
Chemical Substances	B7-H1 Antigen ; Integrin beta4
Language	English
Publishing date	2022-03-28
Publishing country	United States
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, N.I.H., Extramural
ZDB-ID	218154-x
ISSN	1540-8140 ; 0021-9525
ISSN (online)	1540-8140
ISSN	0021-9525
DOI	10.1083/jcb.202108083
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Article: Comparative Pharmacokinetic Study of Three Major Bioactive Components in the Normal and Different Courses of Liver-Stagnation and Spleen-Deficiency Syndrome Depressive Rats after Intragastric Administration of Zhi-Zi-Hou-Po Decoction

Luo, Kaiwen / Wang, Mengdie / Pan, Xiangxiang / Xing, Yadong

Journal of Analytical Methods in Chemistry (Online). 2022 Mar. 24, v. 2022

2022

Abstract	To reveal the pharmacokinetic process of narirutin, naringin, and honokiol in normal and different courses of liver-stagnation and spleen-deficiency syndrome depressive rats after intragastric administration of Zhi-Zi-Hou-Po decoction (ZZHPD), a rapid ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) method was proposed in this study. Chronic unpredictable mild stress (CUMS) rat was employed as the depression model. Acetonitrile solution containing 0.1% acetic acid and methyl alcohol-water (50 : 50, v/v) was chosen as the protein precipitant and redissolve solution severally; a Shim-pack GISS C₁₈ column coupled with 0.1% aqueous acetic acid-acetonitrile in gradient elution was employed to separate the mixed constituents in plasma. The WinNonLin software (version 6.1) was chosen as the analytical tool for the pharmacokinetic parameters. The results indicated that compared with rats in the control group, the sucrose preference, scores in the open-field test, and the concentration of 5-HT in plasma of rats in CUMS and CUMS + ZZHPD treatment groups were lower, while the immobility time in the forced swimming test of rats in these two groups was longer, which implied that the depression model was successful. These behavioral and biochemical indexes of rats above in the CUMS + ZZHPD treatment group were improved after oral administration of ZZHPD, which indicated that the antidepressant effect of ZZHPD was definite. The UHPLC-MS/MS method was stabilized, sensitive, and exclusive, and the extraction recovery and matrix effect of three analytes were all above 89%. The Tₘₐₓ, AUC, and Cₘₐₓ of three ingredients in CUMS-induced depression rats were significantly larger than control rats, while these pharmacokinetic parameters in CUMS + ZZHPD treatment rats were decreased significantly compared with CUMS-induced depression rats, which may relate with the changes in physiological function of the gastrointestinal tract and liver in CUMS-induced liver-stagnation and spleen-deficiency syndrome depressive rats. This study provided important information for the clinical rational use of ZZHPD in antidepressant treatment.
Keywords	acetic acid ; acetonitrile ; antidepressants ; chemical species ; computer software ; digestive tract ; honokiol ; intragastric administration ; liver ; models ; naringin ; oral administration ; pharmacokinetics ; rats ; serotonin ; sucrose ; tandem mass spectrometry ; ultra-performance liquid chromatography
Language	English
Dates of publication	2022-0324
Publishing place	Hindawi
Document type	Article
ZDB-ID	2654178-6
ISSN	2090-8873 ; 2090-8865
ISSN (online)	2090-8873
ISSN	2090-8865
DOI	10.1155/2022/8657616
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Article ; Online: Method for trace determination of N‐nitrosamines impurities in metronidazole benzoate using high‐performance liquid chromatography coupled with atmospheric‐pressure chemical ionization tandem mass spectrometry

Yin, Mingxing / Hu, Yinuo / Fan, Huajun / Wang, Qiulan / Wang, Mengdie / Wang, Wenqing / Shi, Chunyang

Journal of Separation Science. 2023 Mar., v. 46, no. 5 p.e2200225-

2023

Abstract: Genotoxic impurity control has been a great concern in the pharmaceutical industry since the recall of the large round of sartans worldwide in 2018. In these sartans, N‐nitrosamines were the main contaminants in active pharmaceutical ingredients and ... ...

Abstract	Genotoxic impurity control has been a great concern in the pharmaceutical industry since the recall of the large round of sartans worldwide in 2018. In these sartans, N‐nitrosamines were the main contaminants in active pharmaceutical ingredients and formulations. Numerous analytical methods have been developed to detect N‐nitrosamines in food, drugs, and environmental samples. In this study, a sensitive method is developed for the trace determination of N‐nitrosamine impurities in metronidazole benzoate pharmaceuticals using high‐performance liquid chromatography/atmospheric‐pressure chemical ionization tandem mass spectrometry in the multiple reaction monitoring mode. The method was validated regarding system suitability, selectivity, linearity, accuracy, precision, sensitivity, solution stability, and robustness. The method showed good linearity with R² ≥ 0.999 and FMₐₙdₑₗ < Fₜₐb₍₉₅%₎ ranging from 0.33 to 8.00 ng/ml. The low limits of detection of N‐nitrosamines were in the range of 0.22–0.80 ng/ml (0.0014–0.0050 ppm). The low limits of quantification were in the range of 0.33–1.20 ng/ml (0.0021–0.0075 ppm), which were lower than the acceptable limits in metronidazole benzoate pharmaceuticals and indicated the high sensitivity of the method. The recoveries of N‐nitrosamines ranged from 84% to 97%. Thus, this method exhibits good selectivity, sensitivity, and accuracy. Moreover, it is a simple, convenient, and scientific strategy for detecting N‐nitrosamine impurities in pharmaceuticals to support the development of the pharmaceutical industry.
Keywords	atmospheric pressure ; high performance liquid chromatography ; ionization ; liquids ; metronidazole ; mutagens ; nitrosamines ; pharmaceutical industry ; separation ; tandem mass spectrometry
Language	English
Dates of publication	2023-03
Publishing place	John Wiley & Sons, Ltd
Document type	Article ; Online
Note	JOURNAL ARTICLE
ZDB-ID	2047990-6
ISSN	1615-9314 ; 1615-9306
ISSN (online)	1615-9314
ISSN	1615-9306
DOI	10.1002/jssc.202200225
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