[Development and application of SSR markers of Saposhnikovia divaricata based on transcriptome].

Transcriptome sequencing was employed to mine the simple sequence repeat(SSR) locus information of Saposhnikovia divaricata and design specific primers, which aimed to provide a basis for the research on the genetic diversity of S. divaricata germplasm resources. The seed purity, 1 000-seed weight, germination rate, and seed vigor were determined. MISA was used to obtain the SSR locus information from 12 606 unigene longer than 1 kb in the transcriptome database. Forty-three pairs of SSR primers designed in Primer 3 were used to analyze the polymorphism of 28 S. divaricata samples of different sources. The results showed that there were differences in the seed purity, 1 000-seed weight, germination rate, vigor, and seed length and width among S. divaricata samples of different sources. Particularly, the germination rate and seed vigor had significant differences, and HB-ZJK1, NMG-CF4, NMG-BT, NMG-HLE1, and NMG-CF2 had significantly higher 1 000-seed weight, germination rate, and seed vigor than the samples of other sources. Among the 86 233 unigene, 12 606(14.62%) unigene contained 15 958 SSR loci, with one SSR locus every 5 009 bp on average. The SSR loci were mainly single nucleotide and dinucleotide repeats, which were dominated by G/C and TC/AG, respectively. All the primers were screened by using 28 S. divaricata sample from different habitats, and the primers corresponding to the amplification products with clear bands and stable polymorphism were obtained. The clustering results of the biological characteristics and genetic diversity of the 28 S. divaricata samples were basically consistent, and the samples of the same origin(HB-AG1, HB-AG2, HB-ZJK1, and HB-ZJK2) generally gathered together and had close genetic relationship. The SSRs in S. divaricata transcriptome has high frequency, rich types, and high polymorphism, which provides candidate molecular markers for the germplasm identification, genetic map construction, and molecular-assisted breeding.

[Programmed cell death induced by drought stress in sprout tumble of Pinellia ternata].

To further study the mechanism of sprout tumble caused by drought,drought stress was simulating with 30% PEG 6000,physiological,and then the morphological changes of Pinellia ternata cells at different treatment time were detected. The results indicated that,along with the period of drought stress continued,the contents of chlorophyll and water potential were decreased,relative electrical conductivity,contents of soluble sugar and MDA increased. Sprout tumble of P. ternata first occurred on the fourth day during drought stress,large scale of sprout tumble appeared on the eighth day with about 73% of tumble rate. The nuclei exposed to drought stress for 2 days were flattened,lobed,invalidated or irregular in shape and significant showed the apoptotic morphological characteristics. Adenylate transferase( ANT) gene expressions were inhibited by drought,with the rapid increase of Caspase-3 enzyme activity,the cell death rate increased. All this proves that the essence of sprout tumble caused by drought is programmed cell death,which may be a self dormancy protection mechanism of P. ternata against adverse environment.

[Effects of shading on key enzyme genesexpression and accumulation of saponins in Panax japonicus var. major].

To explore the effects of shading and the expression of key enzyme genes on the synthesis and accumulation of Panax japonicus var. major saponins, different shading treatments (0%, 30%,50%) of potted P. japonicus var. major were used as test materials, the expression of three key enzyme genes(CAS,DS,ß-AS) of leaves and rhizomes in different growth periods of P. japonicus var. major was determined by real-time quantitative PCR, the content of total saponins was determined by ultraviolet spectrophotometry. The results indicated that, in flowering stage, CAS,DS,ß-AS were highly expressed in the aerial parts of P. japonicus var. major, 30% shading treatment significantly inhibited the expression of CAS in leaves and promoted the expression of DS and ß-AS in stems, leaves and flowers, it was speculated that the main part of saponin synthesis was leaf in this stage. Both the expression levels of DS and ß-AS and changes in the content of total saponins in leaves showed a tendency of low-high-low throughout the growth cycle, correlation coefficient analysis showed that there was a positive correlation between them. Compared with control, the expression levels of DS and ß-AS and the content of total saponins were greatly enhanced under shading treatment, 30% shading treatment significantly promoted the accumulation of total saponins. Therefore, it is suggested that 30% shading treatment should be applied to the artificial cultivation of P. japonicus var. major, which is beneficial to the accumulation and quality improvement of saponins.

[Flavor and meridian tropism classification analysis of Callianthemum taipaicum].

To explore the flavor and meridian tropism classification of Callianthemum taipaicum by principal components analysis(PCA) and partial least square analysis(PLS). Meanwhile,to establish a high performance liquid chromatography-tandem mass spectrometry(HPLC-ESI-MS) method for the simultaneous determination of 55 active components from 13 kinds of Ranunculaceae of Chinese traditional herbs. Samples were separated on HPLC system by Agilent 5 TC-C18（2）（4.6 mm×250 mm，5 µm）column and eluted with acetonitrile and 0.1% formic acid at the flow rate of 0.6 mL·min⁻¹. The data were performed by HPLC-ESI-MS with multiple reaction monitoring（MRM）scanning mode under positive and negative ion modes and quantified by external standards. The data from 13 Ranunculaceae herbs were analyzed by the PLS-tree and cooman's prediction of PCA and PLS to evaluate the similarities and differences of C. taipaicum in flavor and meridian tropism. The results showed that calibration curves of 55 components all showed good linearity, r>0.99,with good precision, repeatability and stability. After compared to other 12 herbs,PCA and PLS results revealed that the C. taipaicum belonged to lung and bladder meridians while its flavor attributive to pungent,warm in nature. In conclusion,the analysis approach of chemometric calculation combined with multi-components quantification is suitable for the classification of meridian tropism and flavor of Chinese traditional medicine,which can be used for alternative research of rare herbs.

Cardiovascular toxicities following the use of tyrosine kinase inhibitors in hepatocellular cancer patients: a retrospective, pharmacovigilance study.

BACKGROUND: Cardiac adverse events (AEs) are common in tyrosine kinase inhibitors(TKIs). This study explored the cardiac AEs of TKIs through the Food and Drug Administration's Adverse Event Reporting System (FAERS). METHODS: Disproportionality analysis and Bayesian analysis were utilized for data mining of the suspected cardiac AEs of TKIs, based on FAERS data from January 2004 to December 2021. RESULTS: A total of 4708 cardiac AEs reports of sorafenib, regorafenib, lenvatinib, and cabozantinib were identified. Hypertension accounts for the most reported cardiac AE. Lenvatinib appears to induce cardiac failure with the highest signals strength [ROR = 7.7 (3.46,17.17)]. Acute myocardial infarction was detected in lenvatinib [ROR = 7.91 (5.64,11.09)] and sorafenib [ROR = 2.22 (1.74, 2.84)]. Acute coronary syndrome was detected in lenvatinib [ROR = 11.57 (6.84, 19.58)] and sorafenib [ROR = 2.81 (1.87,4.24)]. Atrial fibrillation was detected in sorafenib [ROR = 1.82 (1.55,2.14)] and regorafenib [ROR = 1.36 (1.03,1.81)]. Meanwhile, aortic dissections were detected in sorafenib [ROR = 5.08 (3.31,7.8)] and regorafenib [ROR = 3.39 (1.52,7.56)]. Most patients developed hypertension and cardiac failure within 30 days of initiating TKI treatments. Patients taking lenvatinib had an increased incidence of developing acute coronary syndrome after 180 days of treatment. CONCLUSION: Analysis of FAERS data provides a precise profile on the characteristics of cardiac AEs associated with different TKI regimens. Distinct monitoring and appropriate management are needed in the care of TKI recipients.

[Molecular Genetic Characteristics of Acute Myeloid Leukemia Patients with CBFß-MYH11 Positive].

OBJECTIVE: To explore mutational characteristics of acute myeloid leukemia (AML) patients with CBFß-MYH11+ and analyze the correlation between the mutations and partial clinical characteristics. METHODS: A total of 62 AML patients with CBFß-MYH11+ were included and 51 candidate genes were screened for their mutations using targeted next-generation sequencing (NGS). The exon 12 of NPM1 , FLT3-ITD , and TAD, bZIP domains of CEBPA were detected by genomic DNA-PCR combined with sanger sequencing. RESULTS: Compared with RUNX1-RUNX1T1 + group, the patients with CBFß-MYH11+ showed higher age, peripheral WBC level, initial induced complete remission (CR) rate, more commonly carried chromosomal abnormalities such as +22, and lower deletion ratio of sex chromosome (-X or -Y) (P<0.05). In AML patients with CBFß-MYH11+, the most common mutation was NRAS , followed by KIT, KRAS , and FLT3-TKD . Compared with RUNX1-RUNX1T1+ group, NRAS and FLT3-TKD were more frequently mutated in patients with CBFß-MYH11+ (51.6% vs 18.7%, 17.7% vs 3.8%) (P<0.05). CONCLUSION: The genomic landscape and clinical characteristics of AML patients with CBFß-MYH11+ are different from patients with RUNX1-RUNX1T1 +.

[Effects of puerarin on proliferation and differentiation of umbilical cord mesenchymal stem cells into osteoblasts in vitro].

This study is to investigate the effects of puerarin on the proliferation and differentiation of umbilical cord mesenchymal stem cells (MSCs) into osteoblasts. Umbilical cord MSCs were cultured by tissue adherence and the third passage of cells was used in the experiment. The effect of puerarin on proliferation of umbilical cord MSCs was measured with MTT. The effects of puerarin on umbilical cord MSCs were evaluated by ALP immunohistochemisty and von kossa staining. The OD value decreased with the increase of puerarin concentration. On 7th day, ALP expression of puerarin group was higher than that of control group. On 14th day, ALP staining showed that the positive rate of puerarin group was higher than that of control group. Von kossa staining showed the quantity of calcium nodules was higher in puerarin group than that of control group. Puerarin can promote the umbilical cord MSCs to differentiate into osteoblasts and has an effect on the proliferation of umbilical cord MSCs.

SIRT3 protects bovine mammary epithelial cells from heat stress damage by activating the AMPK signaling pathway.

With global warming, heat stress has become an important challenge for the global dairy industry. Sirtuin 3 (SIRT3), an important mitochondrial NAD+dependent decarboxylase and a major regulator of cellular energy metabolism and antioxidant defense, is integral to maintaining normal mitochondrial function. The aim of this study was to assess the protective effect of SIRT3 on damage to bovine mammary epithelial cells (BMECs) induced by heat stress and to explore its potential mechanism. Our results indicate that SIRT3 is significantly downregulated in heat-stressed mammary tissue and high-temperature-treated BMECs. SIRT3 knockdown significantly increased the expression of HSP70, Bax, and cleaved-caspase 3 and inhibited the production of antioxidases, thus promoting ROS production and cell apoptosis in BMECs. In addition, SIRT3 knockdown can aggravate mitochondrial damage by mediating the expression of genes related to mitochondrial fission and fusion, including dynamin-related protein 1, mitochondrial fission 1 protein, and mitochondrial fusion proteins 1and 2. In addition, SIRT3 knockdown substantially decreased AMPK phosphorylation in BMECs. In contrast, SIRT3 overexpression in high-temperature treatment had the opposite effect to SIRT3 knockdown in BMECs. SIRT3 overexpression reduced mitochondrial damage and weakened the oxidative stress response of BMECs induced by heat stress and promoted the phosphorylation of AMPK. Taken together, our results indicate that SIRT3 can protect BMECs from heat stress damage through the AMPK signaling pathway. Therefore, the reduction of oxidative stress by SIRT3 may be the primary molecular mechanism underlying resistance to heat stress in summer cows.

Quantitative trait loci for resistance to Pyrenophora tritici-repentis race 1 in a Chinese wheat.

Tan spot, caused by Pyrenophora tritici-repentis, is an economically important foliar disease of wheat worldwide. Eight races of the pathogen have been characterized on the basis of their ability to cause necrosis or chlorosis in a set of differential wheat lines. Race 1 produces two host-selective toxins, Ptr ToxA and Ptr ToxC, that induce necrosis and chlorosis, respectively, on leaves of sensitive wheat genotypes. A population of recombinant inbred lines was developed from a cross between Chinese landrace Wangshuibai (resistant) and Chinese breeding line Ning7840 (highly susceptible) to identify chromosome regions harboring quantitative trait loci (QTL) or genes for tan spot resistance. Plants were inoculated at the four-leaf stage in a greenhouse and percent leaf area diseased was scored 7 days after inoculation. Two QTL for resistance to race 1 were mapped to the short arms of chromosomes 1A and 2B in the population. The QTL on 1AS, designated as QTs.ksu-1AS, showed a major effect and accounted for 39% of the phenotypic variation; the QTL on 2BS, designated as QTs.ksu-2BS, explained 4% of the phenotypic variation for resistance. A toxin infiltration experiment demonstrated that both parents were insensitive to Ptr ToxA, suggesting that the population was most likely segregating for reaction to chlorosis, not necrosis. The markers closely linked to the QTL should be useful for marker-assisted selection in wheat-breeding programs.

catena-Poly[[(2,2'-bipyridine)-nickel(II)]-µ-2,4'-oxydibenzoato].

In the title compound, [Ni(C(14)H(8)O(5))(C(10)H(8)N(2))](n), the Ni(II) atom is six-coordinated in a slightly distorted octa-hedral geometry by four O atoms from two chelating carboxyl-ate groups of symmetry-related 2,4'-oxydibenzoate anions and by two N atoms from a 2,2'-bipyridine ligand. The Ni(II) atoms are bridged by the 2,4'-oxydibenzoate anions, resulting in the formation of helical chains parallel to [010] with a repeating unit of 15.039 (2) Å.

[Expression and clinical significance of platelet-derived growth factor A in placenta of pre-eclampsia.].

OBJECTIVE: To investigate the expression and clinical significance of platelet-derived growth factor A (PDGF-A) on placenta tissue from pre-eclampsia. METHODS: The expression of PDGF-A in the placenta of 38 pre-eclampsia patients and 22 normal pregnant women at third trimester was detected by immunohistochemistry method. RESULTS: (1) PDGF-A was mainly expressed in the cytomembrane and cytoplasm of cytotrophoblasts and the endothelial cell of capillary in placenta. (2) The rates of PDGF-A expression of cytotrophoblasts were 63% (24/38) in pre-eclampsia group and 32% (7/22) in normal pregnancy group, which exhibited significant difference (P < 0.05). (3) The rates of PDGF-A expression of endothelial cell were 68% (26/38) in pre-eclampsia group and 27% (6/22) in normal pregnancy group, which also showed significant difference (P < 0.01). (4) The rates of PDGF-A expression of cytotrophoblasts were 39% (7/18) in mild pre-eclampsia patients and 85% (17/20) in severe pre-eclampsia, which reached statistical difference (P < 0.01). CONCLUSION: The increasing expression of PDGF-A in cytotrophoblast and endothelial cell in placenta might confer the occurrence and progression of pre-eclampsia.

[Inhibitory effect of chloroquine on airway hyperresponsiveness in asthmatic mice].

OBJECTIVE: To investigate the effect of chloroquine on airway hyperresponsiveness in asthmatic mice and explore the possible mechanism. METHODS: Balb/c mouse models of asthma established using OVA received intraperitoneal injections of chloroquine, dexamethasone, or both prior to OVA challenge. Within 24 h after the final challenge, airway hyper- responsiveness (AHR) of the mice was assessed, and the total cell count and the counts of different cell populations in the bronchoalveolar lavage fluid (BALF) were determined under light microscopy. The severity of lung inflammation was evaluated using HE staining, and the concentrations of IL-6 and PGF2α in the BALF were detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: Chloroquine pretreatment significantly decreased AHR (P<0.001) in the asthmatic mice and reduced the total cell count (P<0.01), eosinophils (P<0.001), neutrophils (P<0.01), and PGF2α levels in the BALF. Chloroquine combined with low-dose dexamethasone significantly lessened inflammations around the bronchioles (P<0.05) and blood vessels (P<0.01) in the lung tissue, and obviously lowered IL-6 (P<0.05) and PGF2α (P<0.001) in the BALF in the asthmatic mice. CONCLUSION: Chloroquine can inhibit AHR in asthmatic mice and produce better anti-inflammatory effect when combined with dexamethasone for treatment of neutrophilic asthma.

Mysterious abrupt carbon-14 increase in coral contributed by a comet.

A large and sudden increase in radiocarbon ((14)C) around AD 773 are documented in coral skeletons from the South China Sea. The (14)C increased by ~ 15‰ during winter, and remain elevated for more than 4 months, then increased and dropped down within two months, forming a spike of 45‰ high in late spring, followed by two smaller spikes. The (14)C anomalies coincide with an historic comet collision with the Earth's atmosphere on 17 January AD 773. Comas are known to have percent-levels of nitrogen by weight, and are exposed to cosmic radiation in space. Hence they may be expected to contain highly elevated (14)C/(12)C ratios, as compared to the Earth's atmosphere. The significant input of (14)C by comets may have contributed to the fluctuation of (14)C in the atmosphere throughout the Earth's history, which should be considered carefully to better constrain the cosmic ray fluctuation.