Publications by authors named "Long Guo"

134 Publications

Evaluation and Comparison of Bioactive Constituents of Artemisiae argyi Folium Collected at Different Developmental Stages.

J AOAC Int 2020 Aug 10. Epub 2020 Aug 10.

Traditional Chinese Medicine Processing Technology Innovation Center of Hebei Province, Hebei University of Chinese Medicine, Shijiazhuang 050200, China.

Background: Artemisiae argyi Folium (AF) has been used as herbal medicine and a food supplement in China and other Asian countries. There is no report about whether the different developmental stages can influence the bioactive compositions of AF.

Objective: To investigate the dynamic changes of bioactive constituents of AF collected at different developmental stages.

Method: The volatile and nonvolatile bioactive components in AF collected at six different developmental stages were analyzed by gas chromatography-mass spectrometry and (GC-MS) high-performance liquid chromatography (HPLC). Chemometrics analysis, including principal component analysis (PCA) and partial least squares discrimination analysis (PLS-DA), were further performed to compare and discriminate the AF samples based on the analysis results.

Results: The results indicated the bioactive compositions in AF underwent obvious changes during the period of growth. The volatile compositions of AF collected at different stages were the same, while their relative contents were different. Six volatile compounds could be regarded as chemical markers, which were responsible for the intergroup differences. The phenolic profiles of AF at different stages indicated a similar composition when the content levels of the main phenolic compounds were variated.

Conclusions: The comparative results will facilitate better understanding of dynamic changes of bioactive constituents of AF samples collected at different stages and will provide useful information for cultivation and utilization of this herbal medicine.

Highlights: The volatile and nonvolatile bioactive components in AF collected at different developmental stages were evaluated and compared for the first time.
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http://dx.doi.org/10.1093/jaoacint/qsaa105DOI Listing
August 2020

Gypenosides Inhibit Inflammatory Response and Apoptosis of Endothelial and Epithelial Cells in LPS-Induced ALI: A Study Based on Bioinformatic Analysis and in vivo/vitro Experiments.

Drug Des Devel Ther 2021 25;15:289-303. Epub 2021 Jan 25.

Department of Anesthesiology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, People's Republic of China.

Introduction: Severe inflammatory response leads to poor prognosis of acute lung injury (ALI), the role of gypenosides (GPs) on ALI is not fully clear. The study aimed at investigating the effects of GPs on ALI.

Methods: We firstly established LPS-induced ALI mice model. Then, we tested whether GPs contributed to alleviate inflammatory response and lung injury of ALI in vivo. In order to identify specific mechanisms of the phenomenon, we conducted a bioinformatic analysis of LPS-induced ALI mice based on GEO database to identify hub differentially expressed genes (DEGs). PPI network of the DEGs was used to find hub-genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were conducted based on the DAVID database to identify which pathways the genes enriched. Then, we tested whether GPs inhibited lung injury and inflammatory response via the enriched pathways. We also tested whether GPs inhibited the apoptosis of endothelial and epithelial cells secondary to severe inflammation.

Results: We found GPs significantly alleviated lung injury and improved the survival rate of LPS-induced ALI mice in vivo. Bioinformatic analysis identified 20 hub-genes from DEGs, they were mainly enriched in NF-κB and TNF-α pathways. GPs could reduce the lung injury and inflammatory response via inhibiting NF-κB and TNF-α pathways in vivo. Our results indicated that GPs also inhibited inflammatory response of epithelial and endothelial cells via NF-κB and TNF-α pathways in vitro. Severe inflammatory response could also lead to apoptosis of endothelial and epithelial cells. Our results indicated that GPs effectively inhibited the apoptosis of endothelial and epithelial cells.

Conclusion: Our study suggested GPs contributed to alleviated lung injury in vivo and inhibited inflammation and apoptosis of endothelial and epithelial cells in vitro, providing novel strategies for the prevention and therapy for ALI.
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http://dx.doi.org/10.2147/DDDT.S286297DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7846875PMC
January 2021

Expanding the phenotypic spectrum of TNFRSF11A-associated dysosteosclerosis: a case with intracranial extramedullary hematopoiesis.

J Hum Genet 2021 Jan 6. Epub 2021 Jan 6.

Laboratory for Bone and Joint Diseases, RIKEN Center for Integrative Medical Sciences, Tokyo, Japan.

Dysosteosclerosis (DOS) is a rare sclerosing bone dysplasia characterized by osteosclerosis and platyspondyly. DOS is genetically heterogeneous and causally associated with mutations in three genes, SLC29A3, CSF1R, and TNFRSF11A. TNFRSF11A has been known as the causal gene for osteopetrosis, autosomal recessive 7, and is recently reported to cause DOS in three cases, which show a complex genotype-phenotype relationship. The phenotypic spectrum of TNFRSF11A-associated sclerosing bone dysplasia remains unclear and needs to be characterized further in more cases with molecular genetic diagnosis. Here, we report another TNFRSF11A-associated DOS case with a homozygous missense mutation (p.R129C). The mutation effect is different from the previous three cases, in which truncated or elongated RANK proteins were generated in isoform specific manner, thus enriching our understanding of the genotype-phenotype association in TNFRSF11A-associated sclerosing bone dysplasia. Besides DOS, our case presented with intracranial extramedullary hematopoiesis, which is an extremely rare condition and has not been identified in any other sclerosing bone dysplasias with molecular genetic diagnosis. Our findings provide the fourth case of TNFRSF11A-associated DOS and further expand its phenotypic spectrum.
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http://dx.doi.org/10.1038/s10038-020-00891-wDOI Listing
January 2021

[Quality control of Dioscoreae Nipponicae Rhizoma based on fingerprint and quantitative analysis].

Zhongguo Zhong Yao Za Zhi 2020 Oct;45(20):4949-4956

Traditional Chinese Medicine Processing Technology Innovation Center of Hebei Province, College of Pharmacy,Hebei University of Chinese Medicine Shijiazhuang 050200, China.

Dioscoreae Nipponicae Rhizoma, the dried rhizoma of Dioscorea nipponica, has been widely used in traditional Chinese medicines. According to the different of the growth and cultivation patterns, Dioscoreae Nipponicae Rhizoma can be divided into two species, the wild Dioscoreae Nipponicae Rhizoma and the cultivated Dioscoreae Nipponicae Rhizoma. In this paper, an accurate and reliable fingerprint of Dioscoreae Nipponicae Rhizoma was established based on HPLC coupled with evaporative light scattering detector(ELSD). A total of 6 common peaks were marked, and the similarity of the Dioscoreae Nipponicae Rhizoma samples was above 0.950. The results indicated that the established fingerprint could be used for quality evaluation of Dioscoreae Nipponicae Rhizoma. Moreover, an HPLC coupled with ELSD method was developed for simultaneous quantitative analysis of six steroidal saponins, including protodioscin, protogracillin, methyl protodioscin, pseudoprotodioscin, dioscin and gracillin in wild Dioscoreae Nipponicae Rhizoma and cultivated Dioscoreae Nipponicae Rhizoma samples. Furthermore, chemometrics analysis such as principal component analysis and partial least squares discriminant analysis were performed to compare and discriminate wild Dioscoreae Nipponicae Rhizoma and cultivated Dioscoreae Nipponicae Rhizoma samples based on the quantitative data. The results indicated that the contents of steroidal saponins were notably different between the wild and cultivated Dioscoreae Nipponicae Rhizoma, and protodioscin and protogracillin were significant to effectively discriminate the wild and cultivated Dioscoreae Nipponicae Rhizoma samples, and these two compounds could be recognized as chemical markers. In conclusion, this present study might provide useful data and acceptable analysis method for identification and quality evaluation of Dioscoreae Nipponicae Rhizoma.
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http://dx.doi.org/10.19540/j.cnki.cjcmm.20200615.201DOI Listing
October 2020

Mapping lead concentrations in urban topsoil using proximal and remote sensing data and hybrid statistical approaches.

Environ Pollut 2021 Mar 24;272:116041. Epub 2020 Nov 24.

State Key Laboratory of Resources and Environmental Information System, Institute of Geographic Sciences and Natural Resources Research, Chinese Academy of Sciences, Beijing, 100101, China.

Due to rapid urbanization in China, lead (Pb) continues to accumulate in urban topsoil, resulting in soil degradation and increased public exposure. Mapping Pb concentrations in urban topsoil is therefore vital for the evaluation and control of this exposure risk. This study developed spatial models to map Pb concentrations in urban topsoil using proximal and remote sensing data. Proximal sensing reflectance spectra (350-2500 nm) of soils were pre-processed and used to calculate the principal components as landscape factors to represent the soil properties. Other landscape factors, including vegetation and land-use factors, were extracted from time-sequential Landsat images. Two hybrid statistical approaches, regression kriging (RK) and geographically weighted regression (GWR), were adopted to establish prediction models using the landscape factors. The results indicated that the use of landscape factors derived from combined remote and proximal sensing data improved the prediction of Pb concentrations compared with useing these data individually. GWR obtained better results than RK for predicting soil Pb concentration. Thus, joint proximal and remote sensing provides timely, easily accessible, and suitable data for extracting landscape factors.
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http://dx.doi.org/10.1016/j.envpol.2020.116041DOI Listing
March 2021

[Comparative study on changes of ginsenosides and activities of American ginseng before and after steaming].

Zhongguo Zhong Yao Za Zhi 2020 Sep;45(18):4404-4410

College of Pharmacy, Hebei University of Chinese Medicine Shijiazhuang 050200, China Traditional Chinese Medicine Processing Technology Innovation Center of Hebei Province, Hebei University of Chinese Medicine Shijiazhuang 050200, China.

The chemical fingerprints of American ginseng were established by high performance liquid chromatography(HPLC) coupled with similarity evaluation system for chromatographic fingerprint of traditionanl Chinese medicine. The results were analyzed with use of stoichiometry methods(cluster analysis, principal component analysis, and orthogonal partial least squares discriminant analysis), and meanwhile, a preliminary study on the antioxidant and anti-proliferation activity of colorectal cancer cells was conducted. By comparing the fingerprints of American ginseng before and after processing, the contents of five components in the eight ginseno-sides quantified in this paper increased, including ginsenoside Rc, Rg_2, Rb_2, Rb_3 and Rd, respectively, and a new component was produced after steaming. The activity study showed that steamed American ginseng had better antioxidant activity and anti-proliferation activity of colorectal cancer cells than raw American ginseng. The research results show that the steaming method of American ginseng used in this experiment has good stability and reproducibility, and the steaming of American ginseng produces similar changes as artificial red ginseng, which provides a certain reference for expanding the application range of American ginseng.
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http://dx.doi.org/10.19540/j.cnki.cjcmm.20200622.306DOI Listing
September 2020

Efficient detection of copy-number variations using exome data: Batch- and sex-based analyses.

Hum Mutat 2021 Jan 11;42(1):50-65. Epub 2020 Nov 11.

Department of Human Genetics, Yokohama City University Graduate School of Medicine, Yokohama, Japan.

Many algorithms to detect copy number variations (CNVs) using exome sequencing (ES) data have been reported and evaluated on their sensitivity and specificity, reproducibility, and precision. However, operational optimization of such algorithms for a better performance has not been fully addressed. ES of 1199 samples including 763 patients with different disease profiles was performed. ES data were analyzed to detect CNVs by both the eXome Hidden Markov Model (XHMM) and modified Nord's method. To efficiently detect rare CNVs, we aimed to decrease sequencing biases by analyzing, at the same time, the data of all unrelated samples sequenced in the same flow cell as a batch, and to eliminate sex effects of X-linked CNVs by analyzing female and male sequences separately. We also applied several filtering steps for more efficient CNV selection. The average number of CNVs detected in one sample was <5. This optimization together with targeted CNV analysis by Nord's method identified pathogenic/likely pathogenic CNVs in 34 patients (4.5%, 34/763). In particular, among 142 patients with epilepsy, the current protocol detected clinically relevant CNVs in 19 (13.4%) patients, whereas the previous protocol identified them in only 14 (9.9%) patients. Thus, this batch-based XHMM analysis efficiently selected rare pathogenic CNVs in genetic diseases.
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http://dx.doi.org/10.1002/humu.24129DOI Listing
January 2021

The prognostic value of postoperative circulating cell-free DNA in operable hepatocellular carcinoma.

Scand J Gastroenterol 2020 Dec 29;55(12):1441-1446. Epub 2020 Oct 29.

Department of General Surgery, Xiangya Hospital, Central South University, Changsha, China.

Background: Tumor biomarkers are eagerly needed in monitoring the recurrence of operable hepatocellular carcinoma (HCC). Circulating cell-free DNA (cfDNA) is a promising noninvasive molecular biomarker for HCC. The current study aimed to evaluate the clinical significance of the postoperative cfDNA in operable HCC.

Methods: This study enrolled 82 HCC patients from January 2018 to June 2019. All patients underwent liver surgery and were pathologically diagnosed with HCC. Postoperative blood samples were collected from each patient. A fluorometric dsDNA assay was used to measure the concentration of cfDNA. We explore the correlation between cfDNA and recurrence. Kaplan-Meier's curves were used to evaluate the recurrence-free survival (RFS). Univariate and multivariate Cox regression analyses were used for assessing the relative clinical variables in predicting recurrence.

Results: Of the 82 HCC patients, 72 (87%) patients are male and the average age was 52.7 ± 12.8 years. The cfDNA-low and cfDNA-high groups had median recurrence times of 19.5 months and 14.0 months, respectively ( = .023). Multivariate analysis showed that postoperative cfDNA, tumour number and microvascular invasion ( < .050) were independent risk factors for recurrence in operable HCC.

Conclusions: Postoperative cfDNA is still a promising marker to predict prognosis in postoperative HCC patients although prospective and large multicenter clinical study is needed to further validate the relationship between cfDNA and HCC recurrence.
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http://dx.doi.org/10.1080/00365521.2020.1839127DOI Listing
December 2020

The third case of TNFRSF11A-associated dysosteosclerosis with a mutation producing elongating proteins.

J Hum Genet 2020 Oct 9. Epub 2020 Oct 9.

Laboratory for Bone and Joint Diseases, RIKEN Center for Integrative Medical Sciences, Tokyo, Japan.

Dysosteosclerosis (DOS) is a distinct form of sclerosing bone disease characterized by platyspondyly and progressive osteosclerosis. DOS is genetically heterogeneous. Three causal genes, SLC29A3, CSF1R, and TNFRSF11A are reported. TNFRSF11A-associated DOS has been identified in two patients; however, TNFRSF11A is also a causal gene for osteopetrosis, autosomal recessive 7 (OP-AR7). Whole-exome sequencing in a patient with sclerosing bone disease identified novel compound heterozygous variants (c.414_427 + 7del, c.1664del) in TNFRSF11A. We examined the impact of the two variants on five splicing isoforms of TNFRSF11A by RT-PCR. We found that c.1664del resulted in elongated proteins (p.S555Cfs*121, etc.), while c.414_427 + 7del generated two aberrant splicing products (p.A139Wfs*19 and p.E132Dfs*19) that lead to nonsense mediated mRNA decay (NMD). In the previous two cases of TNFRSF11A-associated DOS, their mutations produced truncated TNFRSF11A protein isoforms. The mutations in all three cases thus contrast with TNFRSF11A mutations reported in OP-AR7, which does not generated truncated or elongated TNFRSF11A proteins. Thus, we identified the third case of TNFRSF11A-associated DOS and reinforced the genotype-phenotype correlation that aberrant protein-producing TNFRSF11A mutations cause DOS.
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http://dx.doi.org/10.1038/s10038-020-00831-8DOI Listing
October 2020

An Activatable and Switchable Nanoaggregate Probe for Detecting H S and Its Application in Mice Brains.

Chem Asian J 2020 Nov 6;15(21):3551-3557. Epub 2020 Oct 6.

State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, 210023, P. R. China.

Employing a sequentially activated probe design method, an activatable, switchable and dual-mode probe was designed. This nanoprobe, HSDPP, could be effectively activated by H S to form H-type aggregates with green emission; subsequently, the aggregates could bind to mtDNA to form monomers and the emIssion color switched from green to deep-red. We exploited HSDPP to image exogenous and endogenous H S in living cells. Of note, for the first time, this novel nanoprobe with an optimal partition coefficient value (LogP=1.269) was successfully applied for tracking the endogenous H S upregulation stimulated by cystathionase activator S-adenosyl-L-methionine (SAM) in mice brains. Finally, our work provides an invaluable chemical tool for probing endogenous H S upregulation in vitro/vivo and, importantly, affords a prospective design strategy for developing switchable chemosensors to unveil the relationship between biomolecules and DNA in mitochondria in many promising areas.
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http://dx.doi.org/10.1002/asia.202000964DOI Listing
November 2020

Genetic disorders associated with the RANKL/OPG/RANK pathway.

J Bone Miner Metab 2021 Jan 17;39(1):45-53. Epub 2020 Sep 17.

Laboratory for Bone and Joint Diseases, RIKEN Center for Integrative Medical Sciences, 4-6-1 Minato-ku, Tokyo, 108-8639, Japan.

The RANKL/OPG/RANK signalling pathway is a major regulatory system for osteoclast formation and activity. Mutations in TNFSF11, TNFRSF11B and TNFRSF11A cause defects in bone metabolism and development, thereby leading to skeletal disorders with changes in bone density and/or morphology. To date, nine kinds of monogenic skeletal diseases have been found to be causally associated with TNFSF11, TNFRSF11B and TNFRSF11A mutations. These diseases can be divided into two types according to the mutation effects and the resultant pathogenesis. One is caused by the mutations inducing constitutional RANK activation or OPG deficiency, which increase osteoclastogenesis and accelerate bone turnover, resulting in juvenile Paget's disease 2, Paget disease of bone 2, familial expansile osteolysis, expansile skeletal hyperphosphatasia, panostotic expansile bone disease, and Paget disease of bone 5. The other is caused by the de-activating mutations in TNFRSF11A or TNFSF11, which decrease osteoclastogenesis and elevate bone density, resulting in osteopetrosis, autosomal recessive 2 and 7, and dysosteosclerosis. Here we reviewed the current knowledge about these genetic disorders with paying particular attention to the updating genotype-phenotype association in the TNFRSF11A-caused diseases.
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http://dx.doi.org/10.1007/s00774-020-01148-4DOI Listing
January 2021

A-G Score Associated With Outcomes in Solitary Hepatocellular Carcinoma Patients After Hepatectomy.

Front Oncol 2020 7;10:1286. Epub 2020 Aug 7.

Department of Liver Surgery, Xiangya Hospital, Central South University, Changsha, China.

The study aimed to investigate the clinical significance of preoperative alpha-fetoprotein (AFP) and gamma-glutamyl transferase (GGT) (A-G score) on hepatocellular carcinoma (HCC) patients. A total of 474 solitary HCC patients were included. Survival analysis was evaluated by Kaplan-Meier method. Prognostic factors were analyzed in a multivariate model. The comparison of the predictive value of AFP, GGT, and A-G score was performed by receiver operating characteristic curve (ROC) analysis and decision curve analysis (DCA). Of the 474 patients, 137(28.9%), 241(50.8%), and 96(20.3%) patients were assigned to A-G score 0, 1, and 2, respectively. In multivariate analysis, A-G score, tumor size, microvascular invasion, tumor differentiation, satellite lesion, and state of HBV infection were independently predictive factors for RFS of solitary HCC patients. The A-G score could significantly stratify solitary HCC patients with a distinguished prognosis. The 1-, 3-, and 5-year RFS and OS among patients with A-G score 1 was better than that of patients with A-G score 2 and worse than that of patients with A-G score 0(all < 0.05). Based on the result from the ROC analysis and DCA analysis, the A-G score appeared to be superior to either AFP or GGT alone in the prediction of prognosis of solitary HCC patients. In the subgroup analysis, the A-G score could accurately predict the prognosis of solitary HCC patients without MVI or with liver cirrhosis. Preoperative A-G score could effectively and simply predict prognosis of solitary HCC patients after hepatectomy, especially for those with non-MVI solitary HCC or those with liver cirrhosis.
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http://dx.doi.org/10.3389/fonc.2020.01286DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7427538PMC
August 2020

Comparison of Bioactive Phenolic Compounds and Antioxidant Activities of Different Parts of .

Molecules 2020 Jul 17;25(14). Epub 2020 Jul 17.

Traditional Chinese Medicine Processing Technology Innovation Center of Hebei Province, Hebei University of Chinese Medicine, Shijiazhuang 050200, China.

Herbs derived from genus have been used as traditional medicines and food supplements in China for hundreds of years. is a famous traditional Chinese medicine derived from genus for the treatment of inflammatory disorders and viral infectious diseases. In the present study, the bioactive phenolic chemical profiles and antioxidant activities of flowers, leaves, and roots of were investigated. Firstly, a high performance liquid chromatography method combined with segmental monitoring strategy was employed to simultaneously determine six bioactive phenolic compounds in samples. Moreover, multivariate statistical analysis, including hierarchical clustering analysis, principal component analysis, and partial least squares discriminant analysis were performed to compare and discriminate different parts of based on the quantitative data. The results showed that three phenolic compounds, caftaric acid, caffeic acid, and luteolin, could be regarded as chemical markers for the differences of flowers, leaves, and roots of . In parallel, total phenolic contents, total flavonoid contents and antioxidant activities of different parts of were also evaluated and compared. It is clear that had antioxidant properties, and the antioxidant capacities of different parts of in three antioxidant assays showed a similar tendency: Flowers ≈ leaves > roots, which revealed a positive relationship with their total phenolic and flavonoid contents. Furthermore, to find the potential antioxidant components of , the latent relationships of the six bioactive phenolic compounds and antioxidant activities of were investigated by Pearson correlation analysis. The results indicated caftaric acid and caffeic acid could be the potential antioxidant ingredients of . The present work may facilitate better understanding of differences of bioactive phenolic constituents and antioxidant activities of different parts of and provide useful information for utilization of this herbal medicine.
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http://dx.doi.org/10.3390/molecules25143260DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7397316PMC
July 2020

Ruminal cellulolytic bacteria abundance leads to the variation in fatty acids in the rumen digesta and meat of fattening lambs.

J Anim Sci 2020 Jul;98(7)

State Key Laboratory of Grassland Agro-Ecosystems, Key Laboratory of Grassland Livestock Industry Innovation, Ministry of Agriculture and Rural Affairs, College of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou, P.R. China.

Ruminal cellulolytic bacteria could be a diagnostic tool for determining the subacute rumen acidosis (SARA) risk in individual ruminants; however, a limited number of studies have investigated the effects of the abundance of ruminal cellulolytic bacteria on the fatty acid (FA) composition of the rumen digesta and the muscle of sheep. Thus, the objective of this study was to evaluate the effect of the variation of rumen cellulolytic bacteria on the rumen fermentation, rumen digesta, and muscle FA composition of fattening lambs fed an identical diet. Forty-eight lambs were reared in individual units and fed a high-concentrate diet consisting of 20% forage and 80% concentrate. All lambs were adapted to diets and facilities for 14 d, and sampling was for 63 d. At the end of the experiment, the rumen fluid, rumen digesta, and longissimus dorsi were collected after slaughter for the measurement of volatile fatty acids, ruminal bacterial DNA, rumen digesta, and muscle FAs. The lambs were classified into the lower cellulolytic bacteria (LCB, n = 10) group and the higher cellulolytic bacteria (HCB, n = 10) group according to the abundance of pH-sensitive cellulolytic bacteria (Ruminococcus albus, Ruminococcus flavefaciens, Fibrobacter succinogenes, and Butyrivibrio fibrisolvens) in the rumen. Ruminal acetate concentration was positively correlated with the number of R. flavefaciens, F. Succinogenes, and B. fibrisolvens (P < 0.05, r > 0.296), whereas propionate and valerate concentrations were negatively correlated with the amount of F. succinogenes and B. fibrisolvens (P < 0.05, r > 0.348). Compared with the LCB group, the acetate (P = 0.018) as well as acetate to propionate ratio (P = 0.012) in the HCB group was higher, but the valerate ratio was lower (P = 0.002). The proportions of even-chain FAs and odd- and branched-chain fatty acid in the rumen digesta of lambs with the HCB were higher (P < 0.05), while the polyunsaturated fatty acids decreased than those in the LCB lambs (P < 0.05), but those FA proportions in the meat were similar between the two groups. The proportion of C17:0 in the meat of lambs in the HCB group was lower than that of lambs in the LCB group (P = 0.033). The proportions of conjugated linoleic acid in rumen digesta and meat were both higher in the HCB group than that in the LCB group (P = 0.046). These results indicated that the ruminal cellulolytic bacteria can alter the FA compositions in rumen digesta and further influenced the FA compositions in the meat of sheep.
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http://dx.doi.org/10.1093/jas/skaa228DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7448102PMC
July 2020

[Dynamic changes of volatile components of Qiai from different harvest time based on GC-MS and chemometrics analysis].

Zhongguo Zhong Yao Za Zhi 2020 May;45(10):2417-2424

Traditional Chinese Medicine Processing Technology Innovation Center of Hebei Province, Hebei University of Chinese Medicine Shijiazhuang 050200, China.

Artemisiae Argyi Folium, the dried leaves of Artemisia argyi, has been widely used in traditional Chinese and folk medicines for a long time. Qiai is one of the top-geoherb of Artemisiae Argyi Folium. Qiai contains various bioactive constituents, such as volatile oils, phenolic acids, flavonoids and terpenoids. Phytochemical studies demonstrated that volatile compounds are the main bioactive constituents in Qiai. Try to investigate dynamic changes of volatile components of Qiai from different harvest time and explore the optimum harvest time of Qiai, in this study, the contents of total volatile oils in Qiai collected from five different harvest time were analyzed by steam distillation method. The results showed that the contents of volatile oils of Qiai were higher in the third harvest time(around the Dragon Boat Festival), which is basically consistent with the traditional harvest time. Furthermore, a sensitive method based on gas chromatography-mass spectrometry(GC-MS) was established for qualitative analysis of volatile compounds in Qiai, and a total of thirty volatile compounds were identified. Chemometrics methods including principal component analysis(PCA) and orthogonal partial least-squares discriminate analysis(OPLS-DA) were applied to explore chemical markers and dynamic changes of volatile components in Qiai from different harvest time, and the results indicated that there were obvious differences in the relative contents of volatile compounds of Qiai samples from different harvest time. Eight volatile compounds, including α-terpinene, γ-terpinene, D-camphor, trans-carveol, α-copaene, isobornylisobutyrate, humulene, and caryophyllene oxide were selected as potential chemical markers. Among the eight chemical markers, the relative contents of α-terpinene, γ-terpinene, α-copaene and caryophyllene oxide were higher in the third harvest period(around the Dragon Boat Festival), which is consistent with the contents of total volatile oils. The present study could provide the basis for investigating the optimum harvest time of Qiai, and might be useful for the quality control of this herbal medicine.
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http://dx.doi.org/10.19540/j.cnki.cjcmm.20200219.203DOI Listing
May 2020

Characterization of an aerobic denitrifier Enterobacter cloacae strain HNR and its nitrate reductase gene.

Arch Microbiol 2020 Sep 3;202(7):1775-1784. Epub 2020 May 3.

Key Laboratory of the Three Gorges Reservoir Region's Eco-Environment, Ministry of Education, Chongqing University, Chongqing, 400045, People's Republic of China.

Enterobacter cloacae strain HNR was found to grow well and denitrify aerobically at high NO-N concentrations. When the concentrations of NO-N were 200, 300 and 500 mg/L, the removal efficiencies of NO-N were 83%, 74.5% and 75%, respectively. More importantly, the intermediates accumulation of NO-N and NH-N was not obvious during the aerobic denitrification processes, resulting in a high TN removal of 82%, 74% and 70%, respectively. Meanwhile, strain HNR also presented the ability of heterotrophic nitrification. With initial NH-N concentrations of 20 and 80 mg/L, the NH-N removal efficiency reached 78% and 76%, respectively. The key nitrate reductase enzyme gene relating to denitrification was successfully amplified by polymerase chain reaction (PCR) from strain HNR, and identified it as napA, which encodings the large catalytic subunit A of periplasmic nitrate reductase (NAPA). The sequence analysis of napA indicates that NAPA is a hydrophilic, non-transmembrane protein. The existence of napA might be crucial for strain HNR to denitrify nitrate under aerobic conditions. This study showed prospect to develop novel technology for nitrogen removal by application of E. cloacae strain HNR.
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http://dx.doi.org/10.1007/s00203-020-01887-yDOI Listing
September 2020

[Bridging system for severe comminuted femoral fracture].

Zhongguo Gu Shang 2020 Apr;33(4):332-6

Department of Orthopaedics, Huzhou Gospel Hospital, the 72nd Army Hospital of the PLA, Huzhou 313000, Zhejiang, China.

Objective: To explore the clinical effect of bridging system in the treatment of severe comminuted femoral fracture.

Methods: From March 2016 to October 2018, 50 patients with severe comminuted femoral fracture including 35 males and 15 females, aged 48 to 72(54.6±8.7) years, were admitted. All cases were comminuted fractures of the femoral shaft, 16 with proximal femur fractures and 7 with distal femur fractures. All cases were all unilateral fractures, 23 on the left and 27 on the right. The time from injury to operation was 5 to 60 (26.7±13.3) hours. The cause of injury was traffic accident, 12 cases with high fall, 35 cases fell and 3 cases fell accidentally. The patients were treated with bridge combined internal fixation system, and the operative effect and fracture healing were analyzed.

Results: The operation was successful in all patients. There was no change to other fixed operation. The operation time was (75.8±12.3) min, the amount of bleeding was(356.4±64.8) ml, and there was no serious postoperative complications such as infection, internal fixation displacement, re fracture and nonunion. After 6 to 36 months follow-up, the fracture healing was evaluated by Warden's score. With the extension of observation time, Warden's score gradually increased, and the time of bone healing was(5.5±0.9) months. Harris score and HSS score were used to evaluate the function of hip and knee joint respectively. With the extension of time, Harris score and HSS score increased gradually. Six months after operation, Harris score was 83.5±11.2, HSS score was 79.7±10.5. During the follow-up period, there were no serious complications such as internal fixation displacement, re-fracture, nonunion of fracture and deep vein thrombosis of lower extremity.

Conclusion: The bridge combined internalfixation system has better safety and effectiveness in the treatment of severe comminuted femoral fracture. As long as the requirements of local anatomy and biomechanics are strictly mastered and the operation risks are fully evaluated in combination with imaging, the better fixation effect can be achieved. The operation has less trauma, fewer complications and simple operation, which is believed to have a wider application potential. Due to the limited sample size and follow-up time, no clinical control was set up, the results of the study still need to be further verified by prospective trials.
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http://dx.doi.org/10.12200/j.issn.1003-0034.2020.04.009DOI Listing
April 2020

Leucine improves α-amylase secretion through the general secretory signaling pathway in pancreatic acinar cells of dairy calves.

Am J Physiol Cell Physiol 2020 06 22;318(6):C1284-C1293. Epub 2020 Apr 22.

College of Animal Science and Technology, Northwest A&F University, Yangling, China.

The present study aimed to elucidate the mechanisms by which leucine impacts the secretion of pancreatic enzymes, especially amylase, by studying the proteomics profiles of pancreatic acinar (PA) cells from dairy cows. PA cells, the experimental model, were treated with four concentrations of leucine (0, 0.23, 0.45, and 0.90 mM). The abundance of different proteins in the four leucine treatment groups was detected. Label-free proteomic analysis enabled the identification of 1,906 proteins in all four treatment groups, and 1,350 of these proteins showed common expression across the groups. The primary effects of leucine supplementation were increased ( < 0.05) citrate synthase and ATPase activity, which enlarged the cytosolic ATP pool, and the upregulation of secretory protein 61 (Sec61) expression, which promoted protein secretion. In summary, these results suggest that leucine increases citrate synthase in the TCA cycle and ATPase activity and promotes the Sec signaling pathway to increase the exocrine function of PA cells.
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http://dx.doi.org/10.1152/ajpcell.00396.2019DOI Listing
June 2020

Preoperative evaluation of microvascular invasion with circulating tumour DNA in operable hepatocellular carcinoma.

Liver Int 2020 08 23;40(8):1997-2007. Epub 2020 May 23.

Department of General Surgery, Xiangya Hospital, Central South University, Changsha, Hunan, China.

Background & Aims: Microvascular invasion (MVI) is a critical prognostic factor for operable hepatocellular carcinoma (HCC). This study aimed to explore the performance of circulating tumour DNA (ctDNA) in evaluating MVI status preoperatively.

Methods: Seventy-three HCC patients were enrolled and randomly divided into a training cohort and a validation cohort in a 2:1 ratio, and preoperative blood and surgical tissue samples were obtained. Genomic alterations were analysed using targeted deep sequencing with a 1021-gene panel.

Results: In training cohort, 260 somatic mutations were identified in 40 blood samples (81.6%). CtDNA mutation was verified in paired tissue sample in 39 patients (97.5%). In univariate analysis, ctDNA allele frequency (AF) and largest tumour diameter were associated with the presence of MVI, but ctDNA AF was the only independent risk factor in multivariate analysis. With the cut-off value of 0.83%, ctDNA AF determined the presence of MVI with the sensitivity of 89.7% and specificity of 80.0% in the training cohort, and the sensitivity of 78.6% and the specificity of 81.8% in the validation cohort. In preoperative evaluation, ctDNA AF, AFP level and BCLC staging were associated with recurrence-free survival in both univariate and multivariate analysis.

Conclusions: CtDNA can serve as an independent risk factor of MVI for operable HCC and help determining precise treatment strategies. The integration of ctDNA in the management of operable HCC may achieve better clinical outcomes.
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http://dx.doi.org/10.1111/liv.14463DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7496978PMC
August 2020

CANT1 deficiency in a mouse model of Desbuquois dysplasia impairs glycosaminoglycan synthesis and chondrocyte differentiation in growth plate cartilage.

FEBS Open Bio 2020 06 23;10(6):1096-1103. Epub 2020 Apr 23.

Laboratory of Laboratory Animal Science and Medicine, Co-Department of Veterinary Medicine, Faculty of Agriculture, Iwate University, Morioka, Japan.

Desbuquois dysplasia (DD) type 1 is a rare skeletal dysplasia characterized by a short stature, round face, progressive scoliosis, and joint laxity. The causative gene has been identified as calcium-activated nucleotidase 1 (CANT1), which encodes a nucleotidase that preferentially hydrolyzes UDP to UMP and phosphate. In this study, we generated Cant1 KO mice using CRISPR/Cas9-mediated genome editing. All F0 mice possessing frameshift deletions at both Cant1 alleles exhibited a dwarf phenotype. Germline transmission of the edited allele was confirmed in an F0 heterozygous mouse, and KO mice were generated by crossing of the heterozygous breeding pairs. Cant1 KO mice exhibited skeletal defects, including short stature, thoracic kyphosis, and delta phalanx, all of which are observed in DD type 1 patients. The glycosaminoglycan (GAG) content and extracellular matrix space were reduced in the growth plate cartilage of mutants, and proliferating chondrocytes lost their typical flat shape and became round. Chondrocyte differentiation, especially terminal differentiation to hypertrophic chondrocytes, was impaired in Cant1 KO mice. These findings indicate that CANT1 is involved in the synthesis of GAG and regulation of chondrocyte differentiation in the cartilage and contribute to a better understanding of the pathogenesis of DD type 1.
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http://dx.doi.org/10.1002/2211-5463.12859DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7262921PMC
June 2020

Recapitulating the human segmentation clock with pluripotent stem cells.

Nature 2020 04 1;580(7801):124-129. Epub 2020 Apr 1.

Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.

Pluripotent stem cells are increasingly used to model different aspects of embryogenesis and organ formation. Despite recent advances in in vitro induction of major mesodermal lineages and cell types, experimental model systems that can recapitulate more complex features of human mesoderm development and patterning are largely missing. Here we used induced pluripotent stem cells for the stepwise in vitro induction of presomitic mesoderm and its derivatives to model distinct aspects of human somitogenesis. We focused initially on modelling the human segmentation clock, a major biological concept believed to underlie the rhythmic and controlled emergence of somites, which give rise to the segmental pattern of the vertebrate axial skeleton. We observed oscillatory expression of core segmentation clock genes, including HES7 and DKK1, determined the period of the human segmentation clock to be around five hours, and demonstrated the presence of dynamic travelling-wave-like gene expression in in vitro-induced human presomitic mesoderm. Furthermore, we identified and compared oscillatory genes in human and mouse presomitic mesoderm derived from pluripotent stem cells, which revealed species-specific and shared molecular components and pathways associated with the putative mouse and human segmentation clocks. Using CRISPR-Cas9-based genome editing technology, we then targeted genes for which mutations in patients with segmentation defects of the vertebrae, such as spondylocostal dysostosis, have been reported (HES7, LFNG, DLL3 and MESP2). Subsequent analysis of patient-like and patient-derived induced pluripotent stem cells revealed gene-specific alterations in oscillation, synchronization or differentiation properties. Our findings provide insights into the human segmentation clock as well as diseases associated with human axial skeletogenesis.
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http://dx.doi.org/10.1038/s41586-020-2144-9DOI Listing
April 2020

[Study on difference of chemical constituents of Qiai in different harvest periods].

Zhongguo Zhong Yao Za Zhi 2019 Dec;44(24):5433-5440

College of Pharmacy,Hebei University of Chinese Medicine Shijiazhuang 050200,China.

Artemisiae Argyi Folium,the dried leaves of Artemisia argyi,has been widely used in traditional Chinese and folk medicines for a long time. Qiai is one of the top-geoherb of Artemisiae Argyi Folium. Trying to investigate dynamic changes of chemical components of Qiai in different harvest periods and explore the optimum harvest time of Qiai,in this study,the contents of total flavonoids and total phenolic acids of 36 batches of Qiai collected in 6 different harvest periods were analyzed by ultraviolet-visible spectrophotometry. Furthermore,an HPLC method was applied for simultaneous determination of eight bioactive compounds including six phenolic acids( 5-caffeoylquinic acid,3-caffeoylquinic acid,4-caffeoylquinic acid,3,4-di-O-caffeoylquinic acid,3,5-di-O-caffeoylquinic acid and 4,5-di-O-caffeoylquinic acid) and two flavonoids( jaceosidin and eupatilin) in Qiai samples. The quantitative results indicated that there were some differences in the contents of total flavonoids,total phenolic acids and bioactive compounds of Qiai samples in different harvest periods. The dynamic changes of total flavonoids and total phenolic acids of Qiai in different harvest periods were consistent. The contents of total flavonoids and total phenolic acids of Qiai samples were higher in the third harvest period( around the Dragon Boat Festival),which is basically consistent with the traditional harvest periods. This present study can provide the basis for determining the suitable harvest time of Qiai,and might be useful for the quality evaluation of this herbal medicine.
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http://dx.doi.org/10.19540/j.cnki.cjcmm.20190830.202DOI Listing
December 2019

Methane suppressed dendritic cells activation in trinitrobenzene sulfonic acid induced colitis.

Int Immunopharmacol 2020 Mar 3;82:106360. Epub 2020 Mar 3.

Department of Anesthesiology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 100 Haining Road, Shanghai 200080, China. Electronic address:

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http://dx.doi.org/10.1016/j.intimp.2020.106360DOI Listing
March 2020

Pharmacodynamic study of prasugrel or clopidogrel in non-ST-elevation acute coronary syndrome with CYP2C19 genetic variants undergoing percutaneous coronary intervention (PRAISE-GENE trial).

Int J Cardiol 2020 04 25;305:11-17. Epub 2020 Jan 25.

Division of Cardiology, UCLA Medical Center, Los Angeles, CA, USA.

Background: The CYP2C19*2 or *3 loss-of-function (LOF) allele is associated with high platelet reactivity (HPR) on clopidogrel treatment. East Asians may benefit from a lower dose of prasugrel due to their more potent platelet inhibitory response. The impact of LOF alleles on the pharmacodynamic response to half-dose prasugrel in patients with non-ST-elevation acute coronary syndrome (NSTE-ACS) undergoing percutaneous coronary intervention (PCI) is unknown.

Methods: Seventy patients with the LOF alleles were assigned to half-dose prasugrel (n = 35, 30-mg load followed by 5 mg daily) or clopidogrel (n = 35, 600-mg load followed by 75 mg daily). The primary endpoint was the rate of HPR, defined as VerifyNow-P2Y12 reaction unit (PRU) >235, at 24 h post loading.

Results: Prasugrel achieved a lower PRU compared to clopidogrel after loading (119 [56-175] vs. 245 [189-299]), at 24 h (34 [8-58] vs. 196 [122-244]), and at 30 days (134 [98-189] vs. 203 [144-248]). Prasugrel had a lower rate of HPR after loading (5.7% vs. 57.1%, p <0.001), at 24 h (2.9% vs. 28.6%, p=0.006), and at 30 days (11.4% vs. 34.3%, p=0.004). Prasugrel had a similar rate of optimal platelet reactivity at 30 days (71.4% vs. 60.0%, p=0.450). There was no significant difference in the occurrence of periprocedural myonecrosis within 48 h after PCI with clopidogrel and prasugrel (22.9% vs. 17.1%, p>0.960).

Conclusions: Half-dose prasugrel provided potent platelet inhibition in NSTE-ACS patients that were carriers of the CYP2C19*2 or *3 allele, with a lower rate of HPR. Periprocedural myonecrosis was not significantly different in the 2 groups.
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http://dx.doi.org/10.1016/j.ijcard.2020.01.058DOI Listing
April 2020

Prognostic Nutritional Index and Systemic Immune-Inflammation Index Predict the Prognosis of Patients with HCC.

J Gastrointest Surg 2021 Feb 5;25(2):421-427. Epub 2020 Feb 5.

Department of General Surgery, Xiangya Hospital, Central South University, 87 Xiangya Road, Changsha, 410008, Hunan, China.

Introduction: Systemic nutrition and inflammation are the critical factors in cancer initiation, evolution, and progression. This study aimed to evaluate the prognostic value of the prognostic nutritional index (PNI) and systemic immune-inflammation index (SII) in hepatocellular carcinoma (HCC) patients who underwent liver resection.

Methods: A total of 202 HCC patients met the criteria and were included in the study. The receiver operating characteristic (ROC) curve was used to calculate the optimal PNI and SII cutoff values. The relationship between PNI/SII and clinicopathologic parameters was analyzed. The effect of PNI and SII on recurrence-free survival (RFS) and overall survival (OS) was investigated by Kaplan-Meier curves and Cox proportional hazards models.

Results: The areas under the ROC curve for PNI and SII were 0.64 and 0.58. The ideal preoperative PNI and SII cutoff values were 50.25 and 461.5, respectively. Multivariate Cox regression analysis identified that the PNI (P = 0.001) and tumor diameter (P = 0.018) were significant prognostic markers for RFS, and that the PNI (P = 0.049), SII (P = 0.039) and tumor diameter (P = 0.001) were significant prognostic markers for OS. The median RFS in the PNI-low and PNI-high groups was 13.5 months and 23 months (P = 0.001), and that in the SII-low and SII-high groups was 18 months and 15 months (P = 0.03), respectively. The median OS in the PNI-low and PNI-high groups was 24 months and 39 months (P = 0.001), and that in the SII-low and SII-high groups was 36 months and 22 months (P = 0.002), respectively.

Conclusion: Interestingly, we found that PNI and SII could be important prognostic parameters for HCC patients who under hepatectomy.
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http://dx.doi.org/10.1007/s11605-019-04492-7DOI Listing
February 2021

The clinical value of total plasma cell-free DNA in hepatitis B virus-related hepatocellular carcinoma.

Ann Transl Med 2019 Nov;7(22):650

Department of General Surgery, Xiangya Hospital, Central South University, Changsha 410008, China.

Background: Circulating cell-free DNA (cfDNA), which is present in the blood, is related to the apoptosis and necrosis of cancer cells; inflammation also influences the total plasma level of cfDNA. However, the total plasma cfDNA level has not been investigated in patients with hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) who experience cancer and HBV infection at the same time. The aim of the study was to investigate total plasma cfDNA in patients with HBV-related HCC.

Methods: HBV-related HCC patients were included from January 2018 to May 2019. All patients underwent hepatectomy and were diagnosed with HCC by histopathology. Peripheral blood samples were obtained preoperatively, and the levels of total plasma cfDNA were quantitated by a fluorometric double-stranded DNA (dsDNA) assay. We examined the correlation between cfDNA and clinical parameters, and recurrence-free survival was evaluated using Kaplan-Meier curves.

Results: Forty-eight HBV-related HCC patients were included. The average age in years was 50.90±13.15, and the mean albumin level was 41.63±5.38 g/L. HBV-DNA, Child-Turcotte-Pugh (CTP) class, TNM stage, tumor number and vascular invasion showed a relationship with total plasma cfDNA (P<0.05), and albumin, prothrombin time (PT) and tumor diameter had linear correlation with plasma cfDNA. Based on multivariate analysis, tumor diameter, vascular invasion, and CTP class (P<0.05) were independent risk factors of total plasma cfDNA. Median recurrence times for low-cfDNA and high-cfDNA groups were 14.729±0.712 and 9.264±1.22 months (P=0.026).

Conclusions: In addition to tumor diameter and vascular invasion, CTP class can influence total plasma cfDNA in HBV-related HCC patients, and the total plasma cfDNA level can be used as a biomarker to predict early recurrence in HBV-related HCC patients.
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http://dx.doi.org/10.21037/atm.2019.10.78DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6944585PMC
November 2019

Genome sequencing in persistently unsolved white matter disorders.

Ann Clin Transl Neurol 2020 01 7;7(1):144-152. Epub 2020 Jan 7.

Division of Neurology, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania.

Genetic white matter disorders have heterogeneous etiologies and overlapping clinical presentations. We performed a study of the diagnostic efficacy of genome sequencing in 41 unsolved cases with prior exome sequencing, resolving an additional 14 from an historical cohort (n = 191). Reanalysis in the context of novel disease-associated genes and improved variant curation and annotation resolved 64% of cases. The remaining diagnoses were directly attributable to genome sequencing, including cases with small and large copy number variants (CNVs) and variants in deep intronic and technically difficult regions. Genome sequencing, in combination with other methodologies, achieved a diagnostic yield of 85% in this retrospective cohort.
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http://dx.doi.org/10.1002/acn3.50957DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6952322PMC
January 2020

[Discussion on medicinal value of Saposhnikoviae Radix based on difference of ketone content in cortex and wood of Saposhnikoviae Radix].

Zhongguo Zhong Yao Za Zhi 2019 Sep;44(18):3948-3953

College of Pharmacy,Hebei University of Chinese Medicine Shijiazhuang 050200,China.

In order to confirm the tradition that bolting Saposhnikoviae Radix could not be used as medicine,the content of four chromone components in the cortex and wood of Saposhnikoviae Radix was analyzed by high performance liquid chromatography( HPLC),and the chemical fingerprints were established,12 common peaks were calibrated. The similarity analysis found that the similarity between batches was 0. 115-0. 995,it indicates that the cortex and wood of Saposhnikoviae Radix have certain differences. On this basis,systematic clustering analysis,principal component analysis and orthogonal partial least squares discriminant analysis were carried out with the content of four chromone components and whether they met the pharmacopoeia criteria as the original variables. The results showed that the content of the four components in the cortex of Saposhnikoviae Radix was much higher than that in the wood,and the four components detected were able to distinguish the cortex and the wood of Saposhnikoviae Radix. The results of the study reveal the tradition that bolting Saposhnikoviae Radix should not be used as medicine dut to decreased quality.
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http://dx.doi.org/10.19540/j.cnki.cjcmm.20190701.109DOI Listing
September 2019

[Determination of 9 isoflavonoids in Puerariae Lobatae Radix with quantitative analysis of multi-components by single marker].

Zhongguo Zhong Yao Za Zhi 2019 Nov;44(22):4888-4895

College of Pharmacy,Hebei University of Chinese Medicine Shijiazhuang 050200,China.

A method for determination of 9 isoflavones in Puerariae Lobatae Radix was established and the accuracy and feasibility of the method were verified. The relative correction factors of eight isoflavonoids,3'-hydroxy puerarin,puerarinapioside,3'-methoxy puerarin,puerarin 6″-O-xyloside,daidzin,genistin,formononetin and daidzein were determined by HPLC method with puerarin as the internal standard. The contents of 9 isoflavonoids in 11 batches of samples were determined by external standard method and QAMS.The accuracy and feasibility of the methods were evaluated by comparison of the quantitative results between external standard method and QAMS. The reproducibility of the relative correction factors was good under different experimental conditions,and there was no significant difference between the external standard method of the 9 compounds and the content of QAMS method. The results showed that using puerarin as an internal standard to simultaneously determine the 8 isoflavonoids mentioned above is accurate and feasible. Thus,it can be used as quality control of Puerariae Lobatae Radix.
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http://dx.doi.org/10.19540/j.cnki.cjcmm.20190812.202DOI Listing
November 2019

Crim1 mutant mice reveal the importance of cysteine 140 in the internal region 1 of CRIM1 for its physiological functions.

Mamm Genome 2019 12 27;30(11-12):329-338. Epub 2019 Nov 27.

Artificial Intelligence Research Center (AIRC), National Institute of Advanced Industrial Science and Technology (AIST), 2-4-7 Aomi, Koto-ku, Tokyo, 135-0064, Japan.

Cysteine-rich transmembrane bone morphogenetic protein regulator 1 (CRIM1) is a type I transmembrane protein involved in the organogenesis of many tissues via its interactions with growth factors including BMP, TGF-β, and VEGF. In this study, we used whole-exome sequencing and linkage analysis to identify a novel Crim1 mutant allele generated by ENU mutagenesis in mice. This allele is a missense mutation that causes a cysteine-to-serine substitution at position 140, and is referred to as Crim1. In addition to the previously reported phenotypes in Crim1 mutants, Crim1 homozygous mice exhibited several novel phenotypes, including dwarfism, enlarged seminal vesicles, and rectal prolapse. In vitro analyses showed that Crim1 mutation affected the formation of CRIM1 complexes and decreased the amount of the overexpressed CRIM1 proteins in the cell culture supernatants. Cys140 is located in the internal region 1 (IR1) of the N-terminal extracellular region of CRIM1 and resides outside any identified functional domains. Inference of the domain architecture suggested that the Crim1 mutation disturbs an intramolecular disulfide bond in IR1, leading to the protein instability and the functional defects of CRIM1. Crim1 highlights the functional importance of the IR1, and Crim1 mice should serve as a valuable model for investigating the functions of CRIM1 that are unidentified as yet.
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http://dx.doi.org/10.1007/s00335-019-09822-3DOI Listing
December 2019