Publications by authors named "Yuan Fang"

1,730 Publications

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Bioequivalence and Pharmacokinetic Study of 2 Edoxaban Tablets in Healthy Chinese Subjects.

Clin Pharmacol Drug Dev 2022 Aug 11. Epub 2022 Aug 11.

Nanjing Clinical Tech Laboratories Inc., Nanjing, Jiangsu, China.

The aim was to investigate the bioequivalence of 2 orally administered edoxaban 60-mg tablets and the food effects on the pharmacokinetics of edoxaban. Sixty-four healthy Chinese subjects participated in this open-label, randomized, 2-sequence, 4-period, crossover study. All subjects randomly received 60-mg generic (test) or branded (reference) edoxaban tablet in each period. Plasma edoxaban concentrations were determined using liquid chromatography-tandem mass spectrometry. The pharmacokinetic parameters maximum concentration (C ) and area under the concentration-time curve (AUC) were compared to assess bioequivalence. The geometric least-squares mean ratios for C , AUC from time 0 to the last measurable time point (AUC ), and AUC from time 0 extrapolated to infinity (AUC ) were 97.0%, 95.4%, and 96.1%, respectively, in the fasting test, and 98.6%, 100.0%, and 99.8%, respectively, in the fed test. Food increased exposure and prolonged the time to maximum concentration of edoxaban. Both formulations displayed comparable safety profiles, with no serious adverse events reported. The 2 products of edoxaban tablets are bioequivalent and safe in healthy Chinese volunteers. Food may have a modest effect on the pharmacokinetic properties of edoxaban.
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http://dx.doi.org/10.1002/cpdd.1156DOI Listing
August 2022

Savolitinib monotherapy exerted significant benefit in a non-small cell lung cancer patient with osimertinib resistance harboring primary EGFR L858R mutation and MET amplification: a case report.

Anticancer Drugs 2022 Aug 10. Epub 2022 Aug 10.

Department of Oncology, The First Affiliated Hospital of Nanjing Medical University and.

Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) have significantly improved response in all stages of patients with EGFR positive mutations in nonsmall cell lung cancer. However, the primary resistance mechanism of EGFR-TKIs has not been thoroughly revealed. Here, we described a case of a 64-year-old male with lung adenocarcinoma presented primary resistance on osimertinib combined with bevacizumab and platinum-based chemotherapy, next-generation sequencing revealed EGFR exon 21 L858R mutation and MET gene amplification. Afterward, savolitinib monotherapy was started until now, and the treatment was temporarily successful, the last follow-up clinical evaluation was near complete response, the progression-free survival has over 7 months. Our case highlights that EGFR-TKIs may be not the optimal choice for lung adenocarcinoma with primary EGFR-sensitive mutation with MET amplification simultaneously, whereas MET inhibitor alone may be an effective treatment option. In clinical practice, we should fully consider the possibility of primary resistance in EGFR-TKIs administration.
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http://dx.doi.org/10.1097/CAD.0000000000001370DOI Listing
August 2022

Anticancer Effect of Puerarin on Ovarian Cancer Progression Contributes to the Tumor Suppressor Gene Expression and Gut Microbiota Modulation.

J Immunol Res 2022 28;2022:4472509. Epub 2022 Jul 28.

Zhejiang Chinese Medicine Museum, Zhejiang Chinese Medical University, Hangzhou City, 310053 Zhejiang Province, China.

Ovarian cancer (OC) causes more deaths than any other cancer of the female reproductive system due to its late presentation and malignant nature. Although significant progress has been made in the diagnosis and treatment of OC over the last decade, chemotherapeutic drug resistance and cancer recurrence remain serious challenges in OC management. In the field of cancer therapy, traditional Chinese herbal medicines and their active compounds have been widely reported to have favorable therapeutic effects on cancer. Recent studies have also revealed the protective effect of puerarin in cancer, but the exact role and underlying mechanism of puerarin in OC remain unclear. Here, we established and OC models to evaluate the anticancer effect of puerarin. It was found that puerarin significantly inhibited OC cell viability and proliferation and induced cell apoptosis. In OC model mice, puerarin treatment suppressed tumor formation and modulated the gut microbiome. In addition, the expression of tumor suppressor genes was activated by puerarin and . These findings add to the existing knowledge on the usefulness of herbal active ingredients for the prevention and treatment of OC and provide a new perspective regarding the therapeutic potential of puerarin in cancer.
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http://dx.doi.org/10.1155/2022/4472509DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9352477PMC
August 2022

Comprehensive analysis of complement-associated molecular features in hepatocellular carcinoma.

Acta Biochim Biophys Sin (Shanghai) 2022 Jul 25. Epub 2022 Jul 25.

Department of Liver Surgery and Transplantation, Liver Cancer Institute, Zhongshan Hospital, Fudan University, and Key Laboratory of Carcinogenesis and Cancer Invasion of Ministry of Education, Shanghai 200032, China.

The complement cascade plays a "complementing" role in human immunity. However, the potential roles of complement system in impacting molecular and clinical features of hepatocellular carcinoma (HCC) remain unclear. In this study, eleven public datasets are analyzed to compare the complement status between normal and cancerous samples based on 18 classical complement-associated genes. The complement scores are constructed to quantify complement signatures of individual tumors. HCC patients in the The Cancer Genome Atlas (TCGA) cohort are focused to perform systematical analyses between complement status and immune infiltration, miRNA expression, DNA methylation, clinicopathological features, and drug response. The results show that the complement scores in normal tissues are dramatically higher than those of tumor tissues. Tumor samples in the TCGA cohort are classified into complement score-low and score-high groups. Pathway analysis reveals that tumor-promoting pathways are typically inhibited in complement score-high group. This study also shows that tumor-killing immune cells, such as CD8 T cells and natural killer cells are abundant and tumor-suppressing miRNAs are upregulated in complement score-high samples. In addition, we identify that complement scores are negatively correlated with certain clinical features, including pathological grade, clinical-stage, and portal vein invasion. Moreover, various molecular features together with complement scores are found to be correlated with response to anti-cancer drugs. This study provides a comprehensive and multidimensional analysis conducive to understanding the role of complement in cancer.
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http://dx.doi.org/10.3724/abbs.2022097DOI Listing
July 2022

Formation mechanism of binary complex based on β-lactoglobulin and propylene glycol alginate with different molecular weights: Structural characterization and delivery of curcumin.

Front Nutr 2022 19;9:965600. Epub 2022 Jul 19.

Particle and Interfacial Technology Group, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.

The complexation of protein and polysaccharide has shown considerable potential for the encapsulation of functional food components. In this work, propylene glycol alginate (PGA) molecules with different molecular weights (100, 500, and 2,000 kDa) were prepared through HO oxidation, which were further combined with β-lactoglobulin nanoparticles (β-lgNPs) to form PGA-β-lgNPs complexes for the delivery of curcumin (Cur). Results showed that the depolymerization of PGA molecule was resulted from the breakage of glycosidic bonds in the main chain, and the depolymerization rate of PGA molecule depended on the reaction time, temperature, solution pH and HO concentration. As the increasing molecular weight of PGA, the particle size, zeta-potential and turbidity of the complexes were obviously increased. The formation of PGA/β-lgNPs complexes was mainly driven by non-covalent interaction, including electrostatic gravitational interaction, hydrogen bonding and hydrophobic effect. Interestingly, the difference in the molecular weight of PGA also led to significantly differences in the micro-morphology of the complexes, as PGA with a high molecular weight (2,000 kDa) generated the formation of a "fruit-tree" shaped structure, whereas PGA with relatively low molecular weight (100 and 500 kDa) led to spherical particles with a "core-shell" structure. In addition, the incorporation of PGA molecules into β-lgNPs dispersion also contributed to the improvement in the encapsulation efficiency of Cur as well as physicochemical stability of β-lgNPs, and PGA with a higher molecular weight was confirmed with a better effect. Findings in the current work may help to further understand the effect of molecular weight of polysaccharide on the physical and structural properties as well as effectiveness as delivery systems of polysaccharide-protein complexes, providing for the possibility for the design and development of more efficient carriers for bioactive compounds in food system.
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http://dx.doi.org/10.3389/fnut.2022.965600DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9344013PMC
July 2022

Childhood trauma, stressful life events, and depression: Exploring the mediating effect of cognitive flexibility.

Psychol Trauma 2022 Aug 4. Epub 2022 Aug 4.

Department of Psychology.

Objective: Childhood trauma and stressful life events have been identified as risk factors for the development of depression. However, little is known about the potential cognitive mechanisms underlying these associations. This study aims to investigate the associations among childhood trauma, stressful life events, and depression and to test a mediating model to examine the roles of cognitive flexibility in these associations.

Method: First-year university students ( = 2627; = 18.29, = .86) were recruited from a medical university in Guangzhou, China. A total of 1,600 (60.9%) females and 1,027 (39.1%) males participated in the survey. Participants were administered self-report measures assessing childhood trauma, stressful life events, cognitive flexibility, and depression.

Results: Childhood trauma and stressful life events were both negative predictors of cognitive flexibility (β = -.32, β = -.19, respectively; s < .01), which in turn was a negative predictor of depression (β = -.50, < .01). Cognitive flexibility partially mediated the relationship between childhood trauma and depression (with the indirect effect = .161, 95% confidence interval, CI [.139, .185]) as well as the relationship between stressful life events and depression (with the indirect effect = .097, 95% CI [.078, .118]).

Conclusions: This study reveals that childhood trauma and stressful life events not only have a direct effect on depression but also increase individuals' risk of depression by decreasing their cognitive flexibility. These findings shed light on the possible potential mechanism underlying the suicide process and highlight the important role of cognitive flexibility in suicide prevention and intervention. (PsycInfo Database Record (c) 2022 APA, all rights reserved).
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http://dx.doi.org/10.1037/tra0001337DOI Listing
August 2022

Human microbiota colonization and pancreatic ductal carcinoma.

Crit Rev Microbiol 2022 Aug 4:1-14. Epub 2022 Aug 4.

Department of General Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease with a high mortality rate and a poor prognosis. The human microbiota has been confirmed to participate in oncogenesis and may influence the treatment response to both chemotherapy and immunotherapy. Evidence for the association of the microbiota with PDAC risk, tumorigenesis, treatment response, and survival period is rapidly emerging. The oral microbiota and gut microbiota have the potential to be used in early diagnosis and risk stratification. Intratumor microbiota-targeted intervention strategies may be used as adjuvants to current treatments to improve therapeutic efficacy and overall survival. Here, we summarize the effect and association of the oral, gut and intratumor microbiota on the oncogenesis, progression and treatment of PDAC, as well as the potential of the microbiota to serve as a biomarker for the diagnosis and prognosis of PDAC, as well as a therapeutic target.
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http://dx.doi.org/10.1080/1040841X.2022.2080526DOI Listing
August 2022

Clinical Investigations of CAR-T Cell Therapy for Solid Tumors.

Front Immunol 2022 18;13:896685. Epub 2022 Jul 18.

Clinical Cancer Center, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.

Cell therapy is a distinguished targeted immunotherapy with great potential to treat solid tumors in the new era of cancer treatment. Cell therapy products include genetically engineered cell products and non-genetically engineered cell products. Several recent cell therapies, especially chimeric antigen receptor (CAR)-T cell therapies, have been approved as novel treatment strategies for cancer. Many clinical trials on cell therapies, in the form of cell therapy alone or in combination with other treatments, in solid tumors, have been conducted or ongoing. However, there are still challenges since adverse events and the limited efficacy of cell therapies have also been observed. Here, we concisely summarize the clinical milestones of the conducted and ongoing clinical trials of cell therapy, introduce the evolution of CARs, discuss the challenges and limitations of these therapeutic modalities taking CAR-T as the main focus, and analyze the disparities in the regulatory policies in different countries.
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http://dx.doi.org/10.3389/fimmu.2022.896685DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9339623PMC
August 2022

Seroprevalence of Kaposi's Sarcoma-Associated Herpesvirus and Risk Factors in Jiuquan area, China.

J Med Virol 2022 Aug 3. Epub 2022 Aug 3.

Department of Microbiology and Parasitology, the Key Laboratory of Microbiology and Parasitology of Anhui Province, the Key Laboratory of Zoonoses of High Institutions in Anhui, School of Basic Medical Sciences, Anhui Medical University, 230032, Hefei, Anhui, China.

The seroprevalence of Kaposi's sarcoma-associated herpesvirus (KSHV) is high in Xinjiang, China. But the seroprevalence of KSHV and risk factors are still unknown in Gansu which is adjacent to Xinjiang. 678 serum samples of general population and 87 serum samples of syphilis patients from Jiuquan, Gansu were tested for antibodies against KSHV including one latent protein (ORF73) and two lytic proteins (ORF65 and K8.1) using the ELISA. The total KSHV-seropositive rate was 15.9% in 678 serum samples of Jiuquan area, the KSHV-seropositive rate of males was higher than females (18.0% VS 14.6%, P > 0.05). The Uygur, Kazakh, Hui, Manchu and Mongolian population had a higher seroprevalence of KSHV than the Han population (43.8%, 40.0%, 34.5%, 30.3%, 35.0% VS 11.0%, respectively) among the ethnic groups in Jiuquan. Compared to the Han, Uygur, Kazak, Hui, Manchu and Mongolian people had an increase in the risk of KSHV of 528.9%, 439.1%, 325.6%, 251.6%, 335.4% (P < 0.001, P < 0.001, P < 0.001, P = 0.002, P = 0.003, respectively). The serum prevalence of KSHV in subjects aged < 20 years, 20-50 years and > 50 years was 13.8%, 14.7% and 20.1%, respectively. Compared to the subjects aged < 20 years, 20-50 years and > 50 years had an increase in the risk of KSHV of 7.4% and 56.9% (P = 0.829 and P = 0.204, respectively). Compared to the positive rate of KSHV in the general population of Anhui, the positive rate of KSHV was significantly higher in the general population of Jiuquan area (15.9% VS 9%, P < 0.01). There was no significant difference in the positive rate of KSHV between the Han population of Jiuquan and the Han population of Anhui (P > 0.05). In the population of syphilis patients of Jiuquan area, the positive rate of KSHV was 30.7%, which was higher than that of the general population in Gansu area (P < 0.05). This study indicates that Gansu has a high seroprevalence of KSHV. Ethnicity and syphilis are risk factors for KSHV infection. This article is protected by copyright. All rights reserved.
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http://dx.doi.org/10.1002/jmv.28045DOI Listing
August 2022

Non-coding RNAs in lung cancer: emerging regulators of angiogenesis.

J Transl Med 2022 08 2;20(1):349. Epub 2022 Aug 2.

Institute of Clinical Medicine, The First Affiliated Hospital, Hengyang Medical School, University of South China, Hengyang, 421001, Hunan, People's Republic of China.

Lung cancer is the second cancer and the leading cause of tumor-related mortality worldwide. Angiogenesis is a crucial hallmark of cancer development and a promising target in lung cancer. However, the anti-angiogenic drugs currently used in the clinic do not achieve long-term efficacy and are accompanied by severe adverse reactions. Therefore, the development of novel anti-angiogenic therapeutic approaches for lung cancer is urgently needed. Non-coding RNAs (ncRNAs) participate in multiple biological processes in cancers, including tumor angiogenesis. Many studies have demonstrated that ncRNAs play crucial roles in tumor angiogenesis. This review discusses the regulatory functions of different ncRNAs in lung cancer angiogenesis, focusing on the downstream targets and signaling pathways regulated by these ncRNAs. Additionally, given the recent trend towards utilizing ncRNAs as cancer therapeutics, we also discuss the tremendous potential applications of ncRNAs as biomarkers or novel anti-angiogenic tools in lung cancer.
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http://dx.doi.org/10.1186/s12967-022-03553-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9344752PMC
August 2022

ALDH2C4 regulates cuticle thickness and reduces water loss to promote drought tolerance.

Plant Sci 2022 Jul 30;323:111405. Epub 2022 Jul 30.

Key Laboratory of Plant Stress Research, College of Life Science, Shandong Normal University, Jinan 250014, Shandong, People's Republic of China. Electronic address:

In Arabidopsis thaliana, ALDH2C4 encodes coniferaldehyde dehydrogenase, which oxidizes coniferaldehyde to ferulic acid. Drought stress is one of the important abiotic stresses affecting plant growth. However, the role of ferulic acid in drought resistance is unknown. To investigate the contribution of ferulic acid to cuticle composition and drought resistance, we used two Arabidopsis aldh2c4 mutant lines. Compared with wild-type (WT) leaves, ferulic acid contents were significantly lower (by more than 50 %) in mutants. The mutants also had lower amounts of cutin and wax, primarily due to reductions in C dioic acid and alkanes, respectively. Furthermore, the leaves of the mutant plants exhibited greater rates of water loss and released chlorophyll faster than WT leaves when immersed in 80 % ethanol, indicating a defective cuticle barrier. The growth of aldh2c4 mutants was severely inhibited, and their leaves showed a higher degree of wilting relative to the WT plants under drought conditions. In aldh2c4 complementation lines, the growth inhibition of the mutant plants under drought stress was alleviated. Taken together, our results demonstrate that ferulic acid plays an important role in the composition and structural properties of the cuticle and that a ferulic acid deficiency in the cutin leads to reduced drought tolerance.
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http://dx.doi.org/10.1016/j.plantsci.2022.111405DOI Listing
July 2022

Three-dimensional printing bilayer membranous nanofiber scaffold for inhibiting scar hyperplasia of skin.

Biomater Adv 2022 Jul 26;138:212951. Epub 2022 May 26.

Medical 3D Printing Innovation Research Center, Department of Orthopaedics, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China; College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620, China. Electronic address:

Hypertrophic scar (HS) is the manifestation of pathological wound healing, which affects the beauty of patients, and even affects the normal physical functions of patients. We aimed to develop a 3D printing layer membranous nanofiber scaffold similar to skin structure. Among them, poly (lactic-co-glycolic acid) copolymer (PLGA) nanofibers were used as the "epidermis" layer above, and a decellular dermis matrix (dECM) nanofiber scaffold was used as the "dermis" layer below. In vitro, experimental results showed that PLGA and dECM nanofiber scaffolds had good biocompatibility. In vivo experiments showed that BLM nanofiber scaffolds could inhibit collagen fiber deposition and angiogenesis, to inhibit the formation of hypertrophic scars. This study shows a simple and effective method for preventing and inhibiting the formation of hypertrophic scars.
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http://dx.doi.org/10.1016/j.bioadv.2022.212951DOI Listing
July 2022

Burden of lung cancer along with attributable risk factors in China from 1990 to 2019, and projections until 2030.

J Cancer Res Clin Oncol 2022 Jul 29. Epub 2022 Jul 29.

Department of Epidemiology and Biostatistics, School of Public Health, Cheeloo College of Medicine, Shandong University, Jinan, China.

Objective: This study aimed to identify and project the epidemiological trends and the burden of lung cancer in China.

Methods: We extracted incidence, mortality, disability-adjusted life-years (DALYs) and age-standardized rates of lung cancer in China, between 1990 and 2019, from the Global Burden of Disease Study (2019). The estimated annual percentage change (EAPC) was applied to quantify the trends of lung cancer burden. Furthermore, we used the Bayesian age-period-cohort model to project the incidence and mortality in the next decade.

Results: From 1990 to 2019, the estimated national number of lung cancer incident cases increased by 224.0% to 832,920, deaths increased by 195.4% to 757,170 and DALYs increased by 146.1% to 17,128,580, respectively. Meanwhile, the ASIR, ASMR and ASDR showed an upward trend (EAPC of 1.33, 0.94 and 0.42, respectively). The ASIR and ASMR among males were about 2 times more than females, but the increase in ASIR in females (EAPC = 2.24) was more obvious than those in males (EAPC = 0.10) from 2020 to 2030. In China, smoking remained responsible for the highest burden of lung cancer, but the contribution of ambient particulate matter pollution to DALYs increased from 10.6% in 1990 to 22.5% in 2019 in total population. Moreover, we predicted that the number of deaths from lung cancer will increase by 42.7% in China by 2030.

Conclusion: In China, the burden of lung cancer has been increasing over the past three decades, which highlights more targeted intervention measures are needed to reduce the burden of lung cancer.
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http://dx.doi.org/10.1007/s00432-022-04217-5DOI Listing
July 2022

Case Report: Duodenal Carcinoma in a 40-Year-Old Asian Man With Cowden Syndrome.

Front Surg 2022 12;9:935048. Epub 2022 Jul 12.

Department of General Surgery, South Hospital of the Zhongshan Hospital/Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.

Introduction: Cowden syndrome is a rare autosomal dominant genetic disease associated with PTEN mutation and is mainly shown as systemic multisystem lesions. The incidence of adenocarcinoma of the duodenum with Cowden syndrome in Asian males is rare. We hereby describe the diagnosis, treatment, and prognosis of a patient with duodenal carcinoma and Cowden syndrome.

Case Description: A 40-year-old Chinese man was hospitalized because of gastrointestinal hemorrhage and anemia due to infiltrating adenocarcinoma of the descending part of the duodenum. He also had typical signs of Cowden syndrome, such as multiple polyps of the gastrointestinal tract, macrocephaly, papilloma of the tongue, soles hyperkeratosis, and melanosis spots. After the pancreaticoduodenectomy (classic Whipple), the lesions revealed the presence of hamartomatoid polyps, and some of them mutated into non-mucinous adenocarcinoma (80%) and mucinous adenocarcinoma (20%). Further investigation showed a lack of PTEN protein expression in the duodenal neoplasm, and genetic analysis showed the mutation of p.E242fs in PTEN. The patient was followed up for 1 year. There was no appearance of recurrence or distant metastasis.

Conclusion: It is suggested that we should pay more attention to the differential diagnosis of duodenal carcinoma combined with gastrointestinal polyps. If multiple gastrointestinal polyps with gastrointestinal bleeding are encountered, Cowden syndrome should be considered, and timely diagnosis and treatment should be implemented.
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http://dx.doi.org/10.3389/fsurg.2022.935048DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9320325PMC
July 2022

The performance of detecting Mycobacterium tuberculosis complex in lung biopsy tissue by metagenomic next-generation sequencing.

BMC Pulm Med 2022 Jul 28;22(1):288. Epub 2022 Jul 28.

Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of USTC, Division of Life Science and Medicine, University of Science and Technology of China, No. 17 Lujiang Road, Hefei, 230001, Anhui, China.

Background: Tuberculosis (TB) is a chronic infectious disease caused by the Mycobacterium tuberculosis complex (MTBC), which is the leading cause of death from infectious diseases. The rapid and accurate microbiological detection of the MTBC is crucial for the diagnosis and treatment of TB. Metagenomic next-generation sequencing (mNGS) has been shown to be a promising and satisfying application of detection in infectious diseases. However, relevant research about the difference in MTBC detection by mNGS between bronchoalveolar lavage fluid (BALF) and lung biopsy tissue specimens remains scarce.

Methods: We used mNGS to detect pathogens in BALF and lung biopsy tissue obtained by CT-guide percutaneous lung puncture (CPLP) or radial endobronchial ultrasound transbronchial lung biopsy (R-EBUS-TBLB) from 443 hospitalized patients in mainland China suspected of pulmonary infections between May 1, 2019 and October 31, 2021. Aim to evaluate the diagnostic performance of mNGS for detecting MTBC and explore differences in the microbial composition in the 2 specimen types.

Results: Among the 443 patients, 46 patients finally were diagnosed with TB, of which 36 patients were detected as MTBC positive by mNGS (8.93%). Striking differences were noticed in the higher detection efficiency of lung biopsy tissue compared with BALF (P = 0.004). There were no significant differences between the 2 specimen types in the relative abundance among the 27 pathogens detected by mNGS from the 36 patients.

Conclusions: This study demonstrates that mNGS could offer an effective detection method of MTBC in BALF or lung tissue biopsy samples in patients suspected of TB infections. When it comes to the situations that BALF samples have limited value to catch pathogens for special lesion sites or the patients have contraindications to bronchoalveolar lavage (BAL) procedures, lung biopsy tissue is an optional specimen for MTBC detection by mNGS. However, whether lung tissue-mNGS is superior to BALF-mNGS in patients with MTBC infection requires further prospective multicenter randomized controlled studies with more cases.
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http://dx.doi.org/10.1186/s12890-022-02079-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9330940PMC
July 2022

Knockout of alleviates traumatic brain injury in mice.

Neural Regen Res 2023 Feb;18(2):350-356

Department of Neurosurgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Jiao Tong University, Shanghai, China.

Sirtuin 2 (SIRT2) inhibition or Sirt2 knockout in animal models protects against the development of neurodegenerative diseases and cerebral ischemia. However, the role of SIRT2 in traumatic brain injury (TBI) remains unclear. In this study, we found that knockout of Sirt2 in a mouse model of TBI reduced brain edema, attenuated disruption of the blood-brain barrier, decreased expression of the nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome, reduced the activity of the effector caspase-1, reduced neuroinflammation and neuronal pyroptosis, and improved neurological function. Knockout of Sirt2 in a mechanical stretch injury cell model in vitro also decreased expression of the NLRP3 inflammasome and pyroptosis. Our findings suggest that knockout of Sirt2 is neuroprotective against TBI; therefore, Sirt2 could be a novel target for TBI treatment.
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http://dx.doi.org/10.4103/1673-5374.346457DOI Listing
February 2023

Identification of Tumor Antigens and Immune Subtypes of Malignant Mesothelioma for mRNA Vaccine Development.

Vaccines (Basel) 2022 Jul 22;10(8). Epub 2022 Jul 22.

Clinical Cancer Center, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China.

Background: mRNA-based cancer vaccines have been considered a promising anticancer therapeutic approach against various cancers, yet their efficacy for malignant mesothelioma (MESO) is still not clear. The present study is designed to identify MESO antigens that have the potential for mRNA vaccine development, and to determine the immune subtypes for the selection of suitable patients.

Methods: A total of 87 MESO datasets were used for the retrieval of RNA sequencing and clinical data from The Cancer Genome Atlas (TCGA) databases. The possible antigens were identified by a survival and a genome analysis. The samples were divided into two immune subtypes by the application of a consensus clustering algorithm. The functional annotation was also carried out by using the DAVID program. Furthermore, the characterization of each immune subtype related to the immune microenvironment was integrated by an immunogenomic analysis. A protein-protein interaction network was established to categorize the hub genes.

Results: The five tumor antigens were identified in MESO. FAM134B, ALDH3A2, SAV1, and RORC were correlated with superior prognoses and the infiltration of antigen-presenting cells (APCs), while FN1 was associated with poor survival and the infiltration of APCs. Two immune subtypes were identified; TM2 exhibited significantly improved survival and was more likely to benefit from vaccination compared with TM1. TM1 was associated with a relatively quiet microenvironment, high tumor mutation burden, and enriched DNA damage repair pathways. The immune checkpoints and immunogenic cell death modulators were also differentially expressed between two subtypes. Finally, FN1 was identified to be the hub gene.

Conclusions: FAM134B, ALDH3A2, SAV1, RORC, and FN1 are considered as possible and effective mRNA anti-MESO antigens for the development of an mRNA vaccine, and TM2 patients are the most suitable for vaccination.
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http://dx.doi.org/10.3390/vaccines10081168DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9331978PMC
July 2022

Epidemiology of Mosquito-Borne Viruses in Egypt: A Systematic Review.

Viruses 2022 07 20;14(7). Epub 2022 Jul 20.

NHC Key Laboratory of Parasite and Vector Biology, National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention (Chinese Center for Tropical Diseases Research), WHO Collaborating Center for Tropical Diseases, National Center for International Research on Tropical Diseases, Shanghai 200025, China.

There are at least five common mosquito-borne viruses (MBVs) recorded in Egypt, including dengue virus (DENV), Rift Valley fever virus (RVFV), West Nile virus (WNV), Chikungunya virus, and Sindbis virus. Unexpected outbreaks caused by MBVs reflect the deficiencies of the MBV surveillance system in Egypt. This systematic review characterized the epidemiology of MBV prevalence in Egypt. Human, animal, and vector prevalence studies on MBVs in Egypt were retrieved from Web of Science, PubMed, and Bing Scholar, and 33 eligible studies were included for further analyses. The monophyletic characterization of the RVFV and WNV strains found in Egypt, which spans about half a century, suggests that both RVFV and WNV are widely transmitted in this nation. Moreover, the seropositive rates of DENV and WNV in hosts were on the rise in recent years, and spillover events of DENV and WNV to other countries from Egypt have been recorded. The common drawback for surveillance of MBVs in Egypt is the lack of seroprevalence studies on MBVs, especially in this century. It is necessary to evaluate endemic transmission risk, establish an early warning system for MBVs, and develop a sound joint system for medical care and public health for managing MBVs in Egypt.
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http://dx.doi.org/10.3390/v14071577DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9322113PMC
July 2022

Effects of Environmental Concentrations of Total Phosphorus on the Plankton Community Structure and Function in a Microcosm Study.

Int J Environ Res Public Health 2022 Jul 9;19(14). Epub 2022 Jul 9.

Key Laboratory of Pollution Process and Environmental Criteria of Ministry of Education and Tianjin Key Laboratory of Environmental Remediation and Pollution Control, College of Environmental Science and Engineering, Nankai University, Tianjin 300071, China.

The excessive nutrients in freshwater have been proven to promote eutrophication and harmful algae blooms, which have become great threats to water quality and human health. To elucidate the responses of the plankton community structure and function to total phosphorus (TP) at environmental concentrations in the freshwater ecosystem, a microcosm study was implemented. The results showed that plankton communities were significantly affected by the TP concentration ≥ 0.1 mg/L treatments. In terms of community structure, TP promoted the growth of Cyanophyta. This effect was transmitted to the zooplankton community, resulting in the promotion of Cladocera growth from day 42. The community diversities of phytoplankton and zooplankton had been continuously inhibited by TP. The principal response curve (PRC) analysis results demonstrated that the species composition of phytoplankton and zooplankton community in TP enrichment treatments significantly ( ≤ 0.05) deviated from the control. For community function, TP resulted in the decline in phytoplankton photosynthesis. The chlorophyll fluorescence parameters were significantly inhibited when TP concentration reached 0.4 mg/L. In TP ≥ 0.1 mg/L treatments, the reductions in total phytoplankton abundances led to a continuous decrease in pH. This study can directly prove that the plankton community changes significantly when TP concentrations are greater than 0.1 mg/L and can help managers to establish specific nutrient management strategies for surface water.
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http://dx.doi.org/10.3390/ijerph19148412DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9324525PMC
July 2022

Parent, child, and situational factors associated with parenting stress: a systematic review.

Eur Child Adolesc Psychiatry 2022 Jul 25. Epub 2022 Jul 25.

Department of Public Health, Erasmus University Medical Centre, Rotterdam, PO Box 2040, 3000 CA, Rotterdam, The Netherlands.

Parenting stress has been related to adverse health outcomes in parents, children, and their families. This systematic review aimed to provide an overview of parental, child, and situational factors related to parenting stress in mothers and fathers. We searched Embase, Medline Epub (Ovid), PsychInfo (Ovid), Web of Science, and Google scholar for studies published between January 1980 and May 2021 evaluating the association between at least one factor and parenting stress. Studies were included only if they reported the association in a general population sample of mothers and fathers with children aged 0-12 years. The parent-child relationship model by Abidin guided the data synthesis. Quality of the evidence was assessed using the Standard Quality Assessment Criteria for Evaluating Primary Research Papers from a variety of fields. In total, 29 studies were included with excellent quality (55%), good (31%), and adequate (14%) methodological quality. There was evidence of an association between maternal depression, child overall problems, child externalizing and internalizing problems, social support, maternal educational level and maternal parenting stress. Evidence was inconsistent for an association between maternal anxiety, family income and maternal parenting stress. There was no evidence of an association for maternal age, child sex and maternal parenting stress. Several modifiable factors (i.e., parental depression and social support) were identified that might guide the development of preventive interventions. Future research should employ longitudinal study designs evaluating protective and risk factors and the pathways that lead to parenting stress, among both fathers and mothers.
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http://dx.doi.org/10.1007/s00787-022-02027-1DOI Listing
July 2022

Ratiometrically pH-Insensitive Upconversion Nanoprobe: Toward Simultaneously Quantifying Organellar Calcium and Chloride and Understanding the Interaction of the Two Ions in Lysosome Function.

Anal Chem 2022 08 24;94(30):10813-10823. Epub 2022 Jul 24.

Shanghai Engineering Research Center of Molecular Therapeutics and New Drug Development, Department of Chemistry, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai 200241, China.

Calcium and chloride levels are closely related to lysosome dysfunction. However, the simultaneous measurement of calcium (Ca) and chloride (Cl) in acidic subcellular organelles, which is conducive to a deep understanding of lysosome-related biological events, remains a challenge. In this study, we developed a pH-insensitive, ratiometric NIR nanoprobe for the simultaneous detection of Ca and Cl in acidic lysosomes and determined the roles of the two ions in lysosome function. The upconversion nanoprobe with blue, green, and red emissions was modified with a Ca-sensitive dye (Rhod-5N) and Cl-responsive fluorophore (10,10'-bis[3-carboxypropyl]-9,9'-biacridinium dinitrate, BAC). As a result of a dual-luminescence resonance energy transfer between upconversion nanoparticles (UCNPs) and Rhod-5N/BAC, the blue and green upconversion luminescence (UCL) of UCNPs were quenched and the red UCL was used as the reference signal. The ratiometric upconversion nanoprobe possesses a specific ability for the concurrent recognition of Ca and Cl ions independent of the influence of the environmental pH. To locate the probe in the lysosome, dextran was further modified with upconversion nanoparticles. Then, the nanoprobe with a high spatial resolution was constructed for the simultaneous monitoring of Ca and Cl in acidic lysosomes. Moreover, it was found that the reduction of lysosomal Cl affects the release of lysosomal Ca, which further blocks the activities of specific lysosomal enzymes. The ratiometric NIR nanoprobe has great potential for decoding and evaluating lysosomal diseases.
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http://dx.doi.org/10.1021/acs.analchem.2c01714DOI Listing
August 2022

The Study on the Clinical Phenotype and Function of HPRT1 Gene.

Child Neurol Open 2022 Jan-Dec;9:2329048X221108821. Epub 2022 Jul 19.

Department of Neurology, Shanghai Children's Hospital, School of medicine, Shanghai Jiao Tong University, Shanghai, China.

Lesch-Nyhan disease (LND) is a rare x-linked purine metabolic neurogenetic disease caused by enzyme hypoxanthine-guanine phosphoriribosyltransferase(HGprt) deficiency, also known as self-destructive appearance syndrome. A series of manifestations are caused by abnormal purine metabolism. The typical clinical manifestations are hyperuricemia, growth retardation, mental retardation, short stature, dance-like athetosis, aggressive behavior, and compulsive self-harm. We identified a point mutation c.151C > T (p. Arg51*) in a pedigree. We analyzed the clinical characteristics of children in a family, and obtained the blood of their parents and siblings for second-generation sequencing. At the same time, we also analyzed and compared the expression of HPRT1 gene and predicted the three-dimensional structure of the protein. And we analyzed the clinical manifestations caused by the defect of the HPRT1 gene. The mutation led to the termination of transcription at the 51st arginine, resulting in the production of truncated protein, and the relative expression of HPRT1 gene in patients was significantly lower than other family members and 10 normal individuals. This mutation leads to the early termination of protein translation and the formation of a truncated HPRT protein, which affects the function of the protein and generates corresponding clinical manifestations.
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http://dx.doi.org/10.1177/2329048X221108821DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9305801PMC
July 2022

[Advances in Intraoperative Neurophysiological Monitoring Techniques in Transnasal Endoscopic Skull Base Surgery].

Sichuan Da Xue Xue Bao Yi Xue Ban 2022 Jul;53(4):579-582

Department of Neurosurgery, West China Hospital, Sichuan University, Chengdu 610041, China.

Transnasal endoscopic skull base surgery has been increasing in volume in recent years and its indications are constantly expanding. The potential occurrence of intraoperative and postoperative neurovascular complications deserves special attention from neurosurgeons. Multimodal intraoperative neurophysiological monitoring technology allows neurosurgeons to monitor cerebral perfusion and the functional status of the associated cranial nerves in real time, thereby enabling surgeons to make prompt adjustments in surgical procedures and strategies and reduce the risks of postoperative neurological complications in patients. Based on available literature, we reviewed how appropriate monitoring strategies were optimized for different key components of transnasal endoscopic skull base procedures, intending to provide reference for clinicians.
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http://dx.doi.org/10.12182/20220760501DOI Listing
July 2022

Pathogenic variants identified using whole-exome sequencing in Chinese patients with primary ciliary dyskinesia.

Am J Med Genet A 2022 Jul 23. Epub 2022 Jul 23.

Shenzhen Institute of Respiratory Disease, The First Affiliated Hospital of Southern University of Science and Technology, The Second Clinical Medical College of Jinan University, Shenzhen People's Hospital, Shenzhen, Guangdong, China.

The genetic factors contributing to primary ciliary dyskinesia (PCD), a rare autosomal recessive disorder, remain elusive for ~20%-35% of patients with complex and abnormal clinical phenotypes. Our study aimed to identify causative variants of PCD-associated pathogenic candidate genes using whole-exome sequencing (WES). All patients were diagnosed with PCD based on clinical phenotype or transmission electron microscopy images of cilia. WES and bioinformatic analysis were then conducted on patients with PCD. Identified candidate variants were validated by Sanger sequencing. Pathogenicity of candidate variants was then evaluated using in silico software and the American College of Medical Genetics and Genomics (ACMG) database. In total, 13 rare variants were identified in patients with PCD, among which were three homozygous causative variants (including one splicing variant) in the PCD-associated genes CCDC40 and DNAI1. Moreover, two stop-gain heterozygous variants of DNAAF3 and DNAH1 were classified as pathogenic variants based on the ACMG criteria. This study identified novel potential pathogenic genetic factors associated with PCD. Noteworthy, the patients with PCD carried multiple rare causative gene variants, thereby suggesting that known causative genes along with other functional genes should be considered for such heterogeneous genetic disorders.
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http://dx.doi.org/10.1002/ajmg.a.62912DOI Listing
July 2022

Return-to-Work Predictions for Chinese Patients With Occupational Upper Extremity Injury: A Prospective Cohort Study.

Front Med (Lausanne) 2022 5;9:805230. Epub 2022 Jul 5.

Shanghai YangZhi Rehabilitation Hospital (Shanghai Sunshine Rehabilitation Centre), School of Medicine, Tongji University, Shanghai, China.

Objective: We created predictive models using machine learning algorithms for return-to-work (RTW) in patients with traumatic upper extremity injuries.

Methods: Data were obtained immediately before patient discharge and patients were followed up for 1 year. K-nearest neighbor, logistic regression, support vector machine, and decision tree algorithms were used to create our predictive models for RTW.

Results: In total, 163 patients with traumatic upper extremity injury were enrolled, and 107/163 (65.6%) had successfully returned to work at 1-year of follow-up. The decision tree model had a lower F1-score than any of the other models (t values: 7.93-8.67, < 0.001), while the others had comparable F1-scores. Furthermore, the logistic regression and support vector machine models were significantly superior to the k-nearest neighbors and decision tree models in the area under the receiver operating characteristic curve (t values: 6.64-13.71, < 0.001). Compared with the support vector machine, logistical regression selected only two essential factors, namely, the patient's expectation of RTW and carrying strength at the waist, suggesting its superior efficiency in the prediction of RTW.

Conclusion: Our study demonstrated that high predictability for RTW can be achieved through use of machine learning models, which is helpful development of individualized vocational rehabilitation strategies and relevant policymaking.
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http://dx.doi.org/10.3389/fmed.2022.805230DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9294147PMC
July 2022

Deep Learning Models for Segmenting Non-perfusion Area of Color Fundus Photographs in Patients With Branch Retinal Vein Occlusion.

Front Med (Lausanne) 2022 30;9:794045. Epub 2022 Jun 30.

Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.

Purpose: To develop artificial intelligence (AI)-based deep learning (DL) models for automatically detecting the ischemia type and the non-perfusion area (NPA) from color fundus photographs (CFPs) of patients with branch retinal vein occlusion (BRVO).

Methods: This was a retrospective analysis of 274 CFPs from patients diagnosed with BRVO. All DL models were trained using a deep convolutional neural network (CNN) based on 45 degree CFPs covering the fovea and the optic disk. We first trained a DL algorithm to identify BRVO patients with or without the necessity of retinal photocoagulation from 219 CFPs and validated the algorithm on 55 CFPs. Next, we trained another DL algorithm to segment NPA from 104 CFPs and validated it on 29 CFPs, in which the NPA was manually delineated by 3 experienced ophthalmologists according to fundus fluorescein angiography. Both DL models have been cross-validated 5-fold. The recall, precision, accuracy, and area under the curve (AUC) were used to evaluate the DL models in comparison with three types of independent ophthalmologists of different seniority.

Results: In the first DL model, the recall, precision, accuracy, and area under the curve (AUC) were 0.75 ± 0.08, 0.80 ± 0.07, 0.79 ± 0.02, and 0.82 ± 0.03, respectively, for predicting the necessity of laser photocoagulation for BRVO CFPs. The second DL model was able to segment NPA in CFPs of BRVO with an AUC of 0.96 ± 0.02. The recall, precision, and accuracy for segmenting NPA was 0.74 ± 0.05, 0.87 ± 0.02, and 0.89 ± 0.02, respectively. The performance of the second DL model was nearly comparable with the senior doctors and significantly better than the residents.

Conclusion: These results indicate that the DL models can directly identify and segment retinal NPA from the CFPs of patients with BRVO, which can further guide laser photocoagulation. Further research is needed to identify NPA of the peripheral retina in BRVO, or other diseases, such as diabetic retinopathy.
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http://dx.doi.org/10.3389/fmed.2022.794045DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9279621PMC
June 2022

Circ_0000181 regulates miR-667-5p/NLRC4 axis to promote pyroptosis progression in diabetic nephropathy.

Sci Rep 2022 Jul 14;12(1):11994. Epub 2022 Jul 14.

Department of Nephrology, The Second Xiangya Hospital of Central South University, No 139 Renmin Road, Changsha, 410011, China.

Our previous research demonstrated that NOD-like receptor family CARD domain-containing protein 4 (NLRC4) inflammasome was overexpressed in renal tissues of patients with diabetic nephropathy (DN). This study further investigated the effect of circRNAs-miRNAs interaction on NLRC4 and their potential mechanisms. DN mice models were first established using STZ. Then, pyroptosis related marker expression was detected using qPCR, western blot (WB), and immunohistochemistry analysis. After that, differentially expressed circRNAs, miRNAs, and mRNAs were investigated using next-generation sequencing. Additionally, the function and potential mechanism of circ_0000181 and miR-667-5p on pyroptosis were measured in vitro DN cell model using MTS, WB, and Enzyme-linked immunosorbent assay. There was an apparent elevation of NLRC4, Caspase1, IL-1β, and IL-18 levels in DN mice. The next-generation sequencing results revealed that there were 947 circRNAs and 390 miRNAs significantly different between the DN and sham kidney tissue, of which circ_0000181 and miR-667-5p had potential targeting effects with NLRC4. Dual-luciferase and functional rescue experiments demonstrated that circ_0000181 promoted NLRC4 inflammasome activation via competitive sponge of miR-667-5p, promoted the release of IL-1β and IL-18, and caused pyroptosis. Altogether, circ_0000181 regulates miR-667-5p/NLRC4 axis to promote pyroptosis progression in DN.
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http://dx.doi.org/10.1038/s41598-022-15607-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9283475PMC
July 2022

Cancer-Associated Fibroblast-Derived Exosomal miRNA-320a Promotes Macrophage M2 Polarization by Regulating PTEN/PI3K Signaling in Pancreatic Cancer.

J Oncol 2022 1;2022:9514697. Epub 2022 Jul 1.

Department of General Surgery, Shanghai Public Health Clinical Center, Zhongshan Hospital (South), Fudan University, Shanghai 200032, China.

Our previous study has indicated that cancer-associated fibroblasts (CAFs) play a crucial role in regulating gemcitabine resistance through transferring exosomal miRNA-106b to cancer cells. Tumor-associated macrophages (TAMs) are recently verified to facilitate gemcitabine resistance. However, the effect of CAFs in regulating TAMs function in pancreatic cancer (PCa) remains unclear. Here, primary CAFs were extracted from tumor tissues of PCa patients, and CAFs-derived exosomes (CAFs-Exo) were acquired and authenticated by transmission electron microscopy, qNano, and western blot analysis. The role of exosomal miRNA-320a in facilitating macrophage M2 polarization was investigated . We found that CAFs-derived conditioned medium (CM) possessed a higher potential to promote macrophage M2 polarization compared with normal fibroblasts (NFs) or PCa cell-derived CM. Furthermore, CAFs-Exo treatment polarized macrophage to M2 phenotype. miRNA-320a levels were remarkably increased in CAFs-Exo versus NFs-Exo. More important, miRNA-320a could be transferred from CAFs to macrophages through exosomes, and miRNA-320a overexpression in macrophages facilitated its M2 polarization. Functionally, miRNA-320a-overexpressed macrophages facilitated PCa cell proliferation and invasion. CAFs pretreated with miRNA-320a inhibitor reduced miRNA-320a expression in CAFs-Exo and led to decreased M2 macrophage polarization. Finally, we verified that miRNA-320a polarized macrophage to M2 phenotype by regulating PTEN/PI3K signaling. Taken together, the current data demonstrated that CAFs-derived exosomal miRNA-320a facilitated macrophage M2 polarization to accelerate malignant behavior of PCa cells.
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http://dx.doi.org/10.1155/2022/9514697DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9270150PMC
July 2022

Progress and Applications of Plant Growth-Promoting Bacteria in Salt Tolerance of Crops.

Int J Mol Sci 2022 Jun 24;23(13). Epub 2022 Jun 24.

Shandong Provincial Key Laboratory of Plant Stress, College of Life Sciences, Shandong Normal University, Ji'nan 250014, China.

Saline soils are a major challenge in agriculture, and salinization is increasing worldwide due to climate change and destructive agricultural practices. Excessive amounts of salt in soils cause imbalances in ion distribution, physiological dehydration, and oxidative stress in plants. Breeding and genetic engineering methods to improve plant salt tolerance and the better use of saline soils are being explored; however, these approaches can take decades to accomplish. A shorter-term approach to improve plant salt tolerance is to be inoculated with bacteria with high salt tolerance or adjusting the balance of bacteria in the rhizosphere, including endosymbiotic bacteria (living in roots or forming a symbiont) and exosymbiotic bacteria (living on roots). Rhizosphere bacteria promote plant growth and alleviate salt stress by providing minerals (such as nitrogen, phosphate, and potassium) and hormones (including auxin, cytokinin, and abscisic acid) or by reducing ethylene production. Plant growth-promoting rhizosphere bacteria are a promising tool to restore agricultural lands and improve plant growth in saline soils. In this review, we summarize the mechanisms of plant growth-promoting bacteria under salt stress and their applications for improving plant salt tolerance to provide a theoretical basis for further use in agricultural systems.
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http://dx.doi.org/10.3390/ijms23137036DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9266936PMC
June 2022
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