Preimplantation genetic testing as a preventive strategy for the transmission of mitochondrial DNA disorders.

Mitochondrial diseases are distinct types of metabolic and/or neurologic abnormalities that occur as a consequence of dysfunction in oxidative phosphorylation, affecting several systems in the body. There is no effective treatment modality for mitochondrial disorders so far, emphasizing the clinical significance of preventing the inheritance of these disorders. Various reproductive options are available to reduce the probability of inheriting mitochondrial disorders, including in vitro fertilization (IVF) using donated oocytes, preimplantation genetic testing (PGT), and prenatal diagnosis (PND), among which PGT not only makes it possible for families to have genetically-owned children but also PGT has the advantage that couples do not have to decide to terminate the pregnancy if a mutation is detected in the fetus. PGT for mitochondrial diseases originating from nuclear DNA includes analyzing the nuclear genome for the presence or absence of corresponding mutations. However, PGT for mitochondrial disorders arising from mutations in mitochondrial DNA (mtDNA) is more intricate, due to the specific characteristics of mtDNA such as multicopy nature, heteroplasmy phenomenon, and exclusive maternal inheritance. Therefore, the present review aims to discuss the utility and challenges of PGT as a preventive approach to inherited mitochondrial diseases caused by mtDNA mutations.

Molecular mechanism of Shengyang Yiwei Decoction in treating chronic kidney disease based on network pharmacology.

There is a serious worldwide health problem caused by chronic kidney disease (CKD), yet there are few viable therapies. Therapeutic promise in the treatment of chronic kidney disease (CKD) has been shown by the use of the traditional Chinese herbal compound Shengyang Yiwei Decoction (SYD). However, the chemical processes through which SYD exerts its effects are still unknown. The purpose of this network pharmacology research is to better understand the molecular mechanism of action of Shengyang Yiwei Decoction (SYD) in the treatment of chronic kidney disease (CKD). Traditional Chinese Medicine Systems Pharmacology (TCMSP) was first searched for information on the chemical components of Shengyang Yiwei Decoction. The molecular targets of SYD were then predicted using the Pharm Mapper service. After that, we used databases like DIG-SEE, TTD, and OMIM to zero down on the targets most closely linked to CKD. Cytoscape 3.2.1 was used to generate the component-target network representing SYD's therapy of CKD. In addition, KEGG signal pathways and GO biological processes were analyzed using the DAVID database, and the findings were displayed via OmicShare Tools. Twenty-two active components were isolated from Shengyang Yiwei Decoction, and they were linked to 36 therapeutic targets for CKD in the current investigation. According to the results of the network pharmacology study, 41 signaling pathways are involved in mediating the therapeutic effects of SYD. In addition, SYD's broad therapeutic impact in CKD therapy was shown to include 29 molecular activities, 14 cell components, and 91 biological processes. This research utilizes a multivariate analysis to provide light on the strategies and outcomes of treating CKD using Shengyang Yiwei Decoction. Clinical therapeutic methods for CKD management may benefit greatly from a thorough knowledge of the underlying processes and material foundation of this disease.

[Textual research on classical prescriptions in Mongolian medicine].

Mongolians have a long history of using prescriptions, which can be classified into four stages as follows: the germination and experience accumulation stage before the 13 th century, the theoretical formation stage from the 13 th to 16 th century, the rapid development stage from the 17 th to 20 th century, and the leaping development stage from the mid-20 th century to the present. The prescriptions from the ancient classical or representative medical books have always been used by Mongolian physicians for generations, and they are still in use due to the definite curative effects. In 2008, the Notice on Issuing the Supplementary Provisions to the Registration and Management of Traditional Chinese Medicine(TCM) described that China has attached more importance to the excavation and development of classical prescriptions. As stipulated in the Law of the People's Republic of China on Traditional Chinese Medicine, the classical prescriptions should be those available in ancient TCM classics and still in wide use, with exact curative effects, distinct features, and obvious advantages. This paper expounded the historical formation and development of classical prescriptions in Mongo-lian medicine, introduced the five most influential ancient medical books revealing the formation and development of these classic prescriptions, and traced the origin of such classical prescriptions as Wenguanmu Siwei Decoction, Shouzhangshen Bawei Decoction, Jianghuang Siwei Decoction and summarized the origin, development history and characteristics of classical prescriptions in Mongolian medicine, aiming to provide a reference for their further research and development.

The complete mitochondrial genome of Aucklandia lappa Decne. (Asteraceae, Aucklandia Falc.).

The complete mitochondrial genome of Aucklandia lappa was sequenced for the first time. The mitochondrial genome length was 320,439 bp, with 45.05% GC contents. There were 67 genes annotated, including 31 known protein-coding genes, 25 tRNAs, and six rRNAs. The maximum likelihood method was used to establish the phylogenetic tree of 37 species. Results have shown that A. lappa and Arctium lappa were sister groups. It reveals the genetic relationship between different species and provides a theoretical basis for the establishment of a classification system.

The complete mitochondrial genome of Aconitum kusnezoffii Rchb. (Ranales, Ranunculaceae).

Aconitum kusnezoffii Rchb. is a medicinal plant in the Ranunculaceae family. In this study, we report the first complete mitochondrial genome of A. kusnezoffii. The total length of the mitochondrial genome of A. kusnezoffii is 440,720 bp and the GC content of 46.85%. The mitochondrial genome contained 37 protein-coding genes, 29 tRNAs, and three rRNAs. These data will provide the basis for the systematic evolutionary analysis of Ranunculaceae.

The complete mitochondrial genome of Glycyrrhiza uralensis Fisch. (Fabales, Leguminosae).

The complete mitochondrial genome of an important medicinal plant Glycyrrhiza uralensis Fisch. is reported for the first time. The mitochondrial genome sequence of G. uralensis was 463,869 bp in length and had a GC content of 45.19%. The genome contained 40 protein-coding genes (PCGs), 30 transfer RNAs (tRNAs), and three ribosomal RNAs (rRNAs). The phylogenetic tree was built based on 25 plants, using the maximum-likelihood method. These data will provide certain help to determine the taxonomic status of G. uralensis.

[Rapid identification of constituents of Urtica hyperborea using UPLC-ESI-Q-TOF-MS/MS method].

This paper deals with the application of ultra-performance liquid chromatography tandem quadrupole time of flight mass spectrometry(UPLC-ESI-Q-TOF-MS/MS) method to rapidly determine and analyze the chemical constituents of methanol extract of Urtica hyperborea. We employed UPLC YMC-Triart C18(2. 1 mm×100 mm,1. 9 µm) column to UPLC analysis with acetonitrile-water(containing 0. 4% formic acid) in gradient as mobile phase. The flow rate was 0. 3 m L·min-1 gradient elution and column temperature was 30℃; the injection volume was 4 µL. ESI ion source was used to ensure the data collected in anegative ion mode. The chemical components of U. hyperborea were identified through retention time,exact relative molecular mass,cleavage fragments of MS/MS and reported data.The results indicated that a total of 31 compounds were identified,including 8 flavonoids,14 phenolic compounds,8 phenylpropanoids(4 coumarins and 4 lignans),and 1 steroidal compound,13 of which were confirmed by comparison. The UPLC-ESI-Q-TOF-MS/MS method could rapid identify the chemical components of U. hyperborea. The above compounds were discovered in U. hyperborea for the first time,which could provide theoretical foundation for further research on the basis of the pharmacodynamics of U. hyperborea.

Heterologous expression of Streptomyces coelicolor trehalose synthase and whole-cell biocatalyst production of trehalose in Escherichia coli / 生物工程学报

The trehalose synthase (ScTreS) gene from Streptomyces coelicolor was successfully cloned and heterologously expressed in Escherichia coli BL21(DE3). The protein purified by Ni-NTA affinity column showed an apparent molecular weight (MW) of 62.3 kDa analyzed by SDS-PAGE. The optimum temperature of the enzyme was 35 °C and the optimum pH was 7.0; the enzyme was sensitive to acidic conditions. By homologous modeling and sequence alignment, the enzyme was modified by site-directed mutagenesis. The relative activities of the mutant enzymes K246A and A165T were 1.43 and 1.39 times that of the wild type, an increased conversion rate of 14% and 10% respectively. To optimize the synthesis conditions of trehalose, the mutant strain K246A was cultivated in a 5-L fermentor and used for whole-cell transformation. The results showed that with the substrate maltose concentration of 300 g/L at 35 °C and pH 7.0, the highest conversion rate reached 71.3%, and the yield of trehalose was 213.93 g/L. However, when maltose concentration was increased to 700 g/L, the yield of trehalose can reach 465.98 g/L with a conversion rate of 66%.

Chemical constituents from Hosta plantaginea flowers / 中成药

AIM To study the chemical constituents from Hosta plantaginea (Lam.) Aschers flowers.METHODS The aqueous extract of H.plantaginea was isolated and purified by Sephadex LH-20 and HPLC column,then the structures of obtained compounds were identified by spectral data.RESULTS Twelve compounds were isolated and identified as 4-hydroxybenzaldehyde (1),4-hydroxylacetophenone (2),5,7-dimethoxy-8-rnethyl-4'-hydroxyflavan (3),5,7-dimethoxy-4'-hydroxyflavan (4),epicatechin (5),catechin (6),epigallocatechin (7),gallocatechin (8),coumaric acid (9),phenethyl-O-β-D-glucoside (10),acetophenone-4-O-β-D-glucoside (11),2-hydroxyl-6-methoxyacetophenone-4-O-β-D-glucoside (12),3,4-dihydroxycinnamyl alcohol-3-O-glucoside (13).CONCLUSION All the compounds are isolated from this plant for the first time.

Differential gene expression profile of miRNAs in mouse models with renal ischemia-reperfusion injury / 器官移植

To screen the differentially-expressed microRNAs (miRNAs) in mouse models with renal ischemia-reperfusion injury (IRI),aiming to offer foundation for unraveling the molecular mechanism of the incidence and progression of IRI.Methods The mouse models with acute IRI were established by renal artery clamping.Fifteen mice were divided into the IRI group and sham surgery group (E group).The animals in the IRI group were subdivided into the A group (45 min ischemia followed by 24 h reperfusion),B group (25 min ischemia followed by 24 h reperfusion),C group (45 min ischemia followed by 4 h reperfusion) and D group (25 min ischemia followed by 4 h reperfusion) (n=3 for each group).The severity ofIRI was evaluated by histological changes and renal function.The differentially-expressed miRNAs in the IRI mouse models at different ischemia time (25 and 45 min) and reperfusion time (4 and 24 h) were screened by using cluster analysis of miRNAs microarray data.The differential expression of miR-695 and miR-145 was validated by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR).Results Both histological changes and renal function confirmed that the IRI mouse models were successfully established.Compared with the sham surgery group,71 differentially-expressed miRNAs were detected in the IRI group including 30 down-regulated miRNAs and 40 up-regulated miRNAs.The results of qRT-PCR demonstrated that if the standardized expression level of miRNAs in the E group was 1,the relative expression levels of miR-695 and miR-145 were 11.82 and 0.31 in the IRI group (both P＜0.05),which were consistent with the chip results.Conclusions After renal IRI,different changes occur in the gene expression profile of miRNAs.These differentially-expressed miRNAs act as molecular biomarkers for renal IRI with potential clinical and scientific research values.

Effect of protein acetylation on hepatitis B virus replication / 临床肝胆病杂志

ObjectiveTo investigate the effect of protein acetylation in host cells on the replication of hepatitis B virus (HBV) in hepatocytes, since HBV infection greatly threatens human health and the acetylation of encoding proteins in infected cells plays an important role in HBV replication and infection. MethodsThe deacetylase inhibitors trichostatin A (TSA) and nicotinamide (NAM) were used to stimulate HBV replication in HepG2.2.15 and HepAD38 cells, and the HBV replication markers were measured. The pan-acetylysin protein and Ac-H3 were examined by Western Blot. ResultsThe stimulation of cells with TSA and NAM increased the overall acetylation level of proteins in cells, and the acetylation level increased in a time- and dose-dependent manner. In the HepG2.2.15 and HepAD38 cells, stimulation with TSA and NAM reduced HBsAg level in the supernatant of cell culture and increased HBV DNA level in a time- and dose-dependent manner, while HBeAg in the supernatant of cell culture and DNA in cells did not change significantly. ConclusionAcetylation of host proteins may be involved in and affect HBV replication in cells, and further analysis and determination of host proteins whose acetylation affects HBV replication in cells help to learn more about the regulation of HBV replication and provide new thoughts for the development of specific antiviral strategies.

[Cloning, expression and bioinformatics analysis of pyruvate dehydrogenase of Echinococcus granulosus].

OBJECTIVE: To clone and express Echinococcus granulosus pyruvate dehydrogenase (EgPDH) gene and analyze EgPDH protein with bioinformatics tools and online database. METHODS: The total RNAs of E. granulosus was extracted and reversely transcribed into cDNA. The EgPDH gene was cloned into pET28b to construct the recombinant vector and expressed in E. coli BL21 (DE3) system subsequently. The signal peptide, transmembrane helices and subcellular location in EgPDH sequence were analyzed by the online software SignalP4.1, TMHMM sever v.2.0 and TargetP1.1, respectively. Subsequently, the structure of EgPDH was predicted by SMART. Finally, the homologue sequence and conserved sites were aligned by using BLASTP and GeneDoc among the homologous sequences of EgPDH. Based on the alignment of PDH sequence, an evolutionary tree of E. granulosus and other species were constructed by the neighbor joining method of MEGA6 software. RESULTS: The EgPDH gene was successfully amplified from cDNA of E. granulosus and expressed in the soluble fractions. The bioinformatics analysis revealed that EgPDH was a classical secreted protein and contained transketolase domain. The homology analysis revealed that the amino acid sequence of EgPDH was highly conserved in catalytic sites Glu57, Leu72, Ile86 and Phe114. The phylogenetic tree analysis of PDH proteins showed the closest relationship between E. granulosus and E. multilocularis. CONCLUSION: An EgPDH gene is cloned and expressed successfully, and the recombinant protein is analyzed by the bioinformatics approaches and structure predication. The study provides useful information for further functional study of the EgPDH protein.

Lignans from Syringa pinnatifolia Hemsl. Var. alashanensis.

Two new lignans, syripinnalignans A and B (1 and 2), together with two known lignans, were isolated from the stem of Syringa pinnatifolia Hemsl. Var. alashanensis. The structures of 1 and 2 were elucidated by spectroscopic methods, including extensive 1D and 2D NMR techniques.

Two new flavonoid glycosides from Artemisia frigida Willd.

An investigation of the n-BuOH-soluble fraction from the aerial parts of Artemisia frigida has led to the isolation of two new flavonoid glycosides, named friginoside A and friginoside B. Their structures were characterized as 5,7-dihydroxy-3',4',5'-trimethoxy flavone 7-O-ß-d-glucuronide (1) and 5,7-dihydroxy-3',4',5'-trimethoxyflavone 7-O-ß-d-glucuronyl-(1 â†’ 2)O-ß-d-glucuronide (2) on the basis of 1D and 2D NMR spectral analysis.

Analysis of the Results of Serum Pre-S1 Antigen and HBV DNA Detection / 华中科技大学学报(医学版)

To study the relationship between serum Pre-S1 antigen and HBV DNA, serum Pre-S1 antigen and HBV DNA marker in 229 patients positive for HbsAg were detected by using ELISA method and compared. It was found that both Pre-S1 antigen and HbeAg were correlated with HBV DNA with the coefficient being 0. 9812 and could be used to reflect the existence or reproduction of HBV DNA well. Although there was statistical correlation between them, their clinical implication was not completely the same. Therefore, a conclusion was drawn that combined detection of Pre-S1 antigen, HBV and HBV DNA in the patients with different kinds of HBV infection was helpful to clinical diagnosis, therapeutic effect evaluation and prognosis judgement.