The impact of age and number of mutations on the size of clonal hematopoiesis.

Clonal hematopoiesis (CH) represents the clonal expansion of hematopoietic stem cells and their progeny driven by somatic mutations. Accurate risk assessment of CH is critical for disease prevention and clinical decision-making. The size of CH has been showed to associate with higher disease risk, yet, factors influencing the size of CH are unknown. In addition, the characteristics of CH in long-lived individuals are not well documented. Here, we report an in-depth analysis of CH in longevous (≥90 y old) and common (60~89 y old) elderly groups. Utilizing targeted deep sequencing, we found that the development of CH is closely related to age and the expression of aging biomarkers. The longevous elderly group exhibited a significantly higher incidence of CH and significantly higher frequency of TET2 and ASXL1 mutations, suggesting that certain CH could be beneficial to prolong life. Intriguingly, the size of CH neither correlates significantly to age, in the range of 60 to 110 y old, nor to the expression of aging biomarkers. Instead, we identified a strong correlation between large CH size and the number of mutations per individual. These findings provide a risk assessment biomarker for CH and also suggest that the evolution of the CH is influenced by factor(s) in addition to age.

[Analysis of dissolution characteristics and content of metal elements in Scutellaria baicalensis].

ICP-OES analytical technique was used to determine the contents of Ca, Mg, Cu, Zn, Fe and Mn in S. baicalensis from different origins, and the dissolution characteristics and the contents of 6 elements were analyzed. There were significant differences in the contents among the prepared medical materials (p < 0.05) and their water extraction also differed significantly. The results showed that there was no relationship on the contents of the 6 elements between in the prepared medical materials and in their water extraction In the prepared medical materials, the contents of the elements were characterized as Mg>Ca>Fe>Mn>Zn>Cu, and the contents in the water extraction was rated from high to low as Mg, Ca, Mn, Zn, Fe and Cu. Most of the measured samples demonstrated lower Ca/Mg, higher Fe/Mn and lower Cu/Zn values, and these ratios were in relation to some efficacy of S. baicalensis, such as anti-hypertensive, anti-tumor activities. The results will provide a scientific basis for further enriching and developing the multi-index system for the quality evaluation.

Two new C-methyl flavanones from the rhizomes and frond bases of Matteuccia struthiopteris.

Two new C-methyl flavanones, (2S)-5,7-dihydroxy-6,8-dimethyl-4'-methoxydihydroflavone-7-O-(6″-O-acetyl)-ß-d-glucopyranoside (1) and (2S)-5,7-dihydroxy-6,8-dimethyldihydroflavone-7-O-(6″-O-acetyl)-ß-d-glucopyranoside (2), together with five known compounds, demethoxymatteucinol-7-O-ß-d-glucopyranoside (3), matteucinol-7-O-ß-d-glucopyranoside (4), 5,7-dihydroxy-6-methyl-4'-methoxydihydroflavone (5), methoxymatteucin (6), and thunberginol C (7), were first isolated from the EtOH extract of the rhizomes and frond bases of Matteuccia struthiopteris. The structures were established by spectral analyses, mainly HR-ESI-MS and 1D and 2D NMR experiments (COSY, HSQC, and HMBC).

[Quality evaluation of Flos Carthami].

OBJECTIVE: To develop methods for qualitative and quantitative analyses of Flos Cartnami from three aspects, pigments, flavonoids and adenosine. METHOD: A method using HPLC coupled with electrochemical detector was developed to determine the content of hydroxysafflor yellow A and fingerprint of Flos Carthami. The chromatographic separation was performed on a Zorbax SB C18 column (4.6 mm x 250 mm, 5 microm) by gradient elution with phosphate buffer and acetonitrile at a flow-rate of 1.0 mL x min(-1), the column temperature was 35 degrees C, the reference electrode was ISAAC (in-situ silver/silver chloride), the working electrode was glassy carbon, the counter electrode was Pt, and the applied potential was + 800 mV. Concentration of adenosine was determined by HPLC-UV on an Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) with water-acetonitrile (95:5) as mobile phase, the flow rate was 1.0 mL x min(-1), the column temperature was 40 degrees C and the detection wavelength was 260 nm. The content of cartharmin was detected using a spectrophotometric method. RESULT: Twenty-one common chromatographic peaks were selected as characteristic peaks in the chromatogram of sample solution of Flos Cartnami. Seven peaks were identified as hydroxysafflor yellow A, 6-hydroxykaempferol-3-O-glucoside, rutin, quercetin-3-O-glucoside, kaempferol-3-O-rutinoside, quercetin, kaempferol. The contents of hydroxysafflor yellow A and adenosine were from 0.35% to 3.58% and from 0.03% per hundred to 0.49% per hundred, respectively. CONCLUSION: The methods can be used to evaluate the quality of Flos Carthami.

[Studies on chemical constituents of rhizome of Matteuccia struthiopteris].

OBJECTIVE: To study the chemical constituents of the rhizome of Matteuccia struthiopteris. METHOD: The constituents were separated and purified by column chromatography with silica gel and Sephadex LH-20. Their structures were identified on the basis of physical and spectral data. RESULT: Six compounds were isolated and identified as demethoxymatteucinol (1), matteucinol (2), pinosylvin (3), matteuorien (4), pinosylvin 3-O-beta-D-glucopyranoside (5), matteuorienate A (6). CONCLUSION: All Compounds were isolated from this plant for the first time.

[Determination of five primary chemical constituents in Cassia angustifolia by HPLC].

OBJECTIVE: To establish a method for determining the content of primary chemical constituents in the leaves of Cassia angustifolia. METHOD: The HPLC with Diamonsil C18 (4.6 mm x 250 mm, 5 microm) column was used , acetonitrile-1% acetic acid (10:90-15: 85-18: 82-20: 80-25: 75) in a gradient manner was used as a mobile phase, with flow rate of 1 mL x min(-1), column temperature at 40 degrees C and detection wavelength at 270 nm. RESULT: The results showed that 5 effective components all separated well and showed good linearity. CONCLUSION: The method was proved to be rapid, sensitive, accurate, credible and repeatable. It can be applied to quality control of Folium Sennae.

[Chemical constituents from the leaves of Cassia angustifolia].

OBJECTIVE: To study the chemical constituents from the leaves of Cassia angustifolia. METHODS: Compounds were isolated and repeatedly purified by chromatographic techniques on silica gel column. Their structures were elucidated by chemical and spectral methods. RESULTS: eight compounds were isolated from the leaves of Cassia angustifolia, and identified as tinnevellin glycoside(I), isorhamnetin-3-O-beta-gentiobioside(II), apigenin-6,8-di-C-glycoside(III), emodin-8-O-beta-D-glucopyranoside(IV), kaempferol(V), aloe emodin(VI), D-3-O-methylinositol(VII), sucrose(VIII). CONCLUSION: Compounds III, VII and VIII are isolated from the plant for the first time.

[Experimental studies on hepatotoxicity of rats induced by Fructus Gardeniae].

OBJECTIVE: To study the hepatotoxicity effects in rats with different extract of Fructus Gardeniae. METHOD: Observe the change of appearance, behavior and weight of rats through oral gavage daily for 3 d. Weigh the liver and calculate the liver index. Detect the ALT, AST and TBIL. Observe the liver tissue by optical microscope. RESULT: The weight and index of liver were increased by 3.08 g x kg(-1) aqueous extract, 1.62 g x kg(-1) alcoholic extract and 0.28 g x kg(-1) geniposide, compared to those of the blank group (P < 0.005, P < 0.001) and the activities of ALT, AST and the content of TBIL were also increased, compared to those of the blank group (P < 0.05, P < 0.001). The liver cells were obviously swell, necrotic and changed with inflammatory infiltrate. CONCLUSION: Aqueous extract, alcoholic extract and geniposide displayed hepatotoxicity, and the geniposide which was the main substance of the Fructus Gardeniae might be mainly responsible for the hepatotoxicity.

[Determining the quantity of hypericin in medicinal materials and asepsis seedings of Hypericum perforatum by HPLC].

OBJECTIVE: To establish the method of determining the quantity of hypericin in Hypericum perforatum and determine the quantity of the hypericin in defferent medicinal materials and asepsis seedings which grow in defferent environment. METHOD: The specimen is extracted with methanol--Pyridine (9:1) ultrasound extraction. Chromatographic assay is performed on a hypersily ODS2 (4.6 mm x 150 mm, 5 microm) column. The mobile phase is composed of methanol -1.56% dihydric natrium phosphate hydrogen natrium solution (shift solution's acidity to 2.1 with phosphoric acid)--ethyl acetate (4:1.9:1), velocity of flow is 1 mL x min(-1); column temperature is 35 degrees C; the detection wavelength is 590 nm. RESULT: A satisfactory seperaration between hypericin and impurity. The calibration curve is linear over the range of 0.0524-0.2620 microg for hypericin (r = 0.9998). The average recovery of hypericin is 97.50%. CONCLUSION: The quantity of hypericin in Hypericum perforatum has something to do with the genetic factor, environment factor, growing period and dry means. The method of determining the quantity of hypericin can be regarded as the method of controling the quantity of medicinal materials.