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.

Clinical studies on the electric automatic massage therapy for recovery of acute sports fatigue.

BACKGROUND: Acute sports fatigue impairs athletes' performance and causes other health issues; therefore, an effective method of relieving acute sports fatigue is being researched. OBJECTIVE: The present study was envisaged to evaluate the effect of electric auto-massage therapy and proprioceptive neuromuscular facilitated (PNF) stretching method on the recovery of acute exercise fatigue using the heart rate variability-based multi-physiological index and RPE scale, and to explore the underlying principle and mechanism. METHOD: Sixty volunteers were divided into the stretching group, massage group and control group (20 subjects each) using the complete randomization method. The massage group chose the kneading, pressing, tapping and patting techniques using the intelligent massage chair to intervene on the volunteers, the stretching group chose the PNF stretching method to intervene on the volunteers, while the control group did not adopt any of these techniques. The Rating Of Perceived Exertion (RPE) score, heart rate (HR), grip strength, skin electrical activity, heart rate variability (HRV) and blood oxygen saturation (SpO2) of the three groups were recorded before and after the intervention. RESULTS: Before the intervention, there was no statistically significant difference between the values of heart rate variability (HRV) in the three groups (P> 0.05), while after the intervention, there was a statistically significant difference between the values of heart rate variability - low frequency/high frequency (HRV (LF/HF)) and HRV (HF) in the three groups as: HRV (HF): ηH2= 0.10; P= 0.022; HRV (LF/HF): ηH2= 0.44; P= 0.001. The results indicated that the different intervention methods presented substantial effects on the values of HRV (HF) and HRV (LF/HF) in the volunteers. The HRV (HF) values of massage group, stretching group and control group were compared, and the difference between the massage group and control group was statistically significant (P= 0.019). Further, the HRV (HF) values of massage group rose more significantly than control group after the intervention, and the difference between HRV (HF) values of massage group and stretching group was not statistically significant. When comparing the HRV (LF/HF) values of massage group, stretching group and control group, the differences between the massage group and stretching group and control group were statistically significant (P= 0.001, P< 0.05), and it was observed that the HRV (LF/HF) values of massage group decreased more significantly than those of stretching group and control group after the intervention. The difference in HRV (LF) values between the three groups after the intervention was not statistically significant (P> 0.05). CONCLUSION: 1. It was observed that the electric automatic massage therapy played a vital role in the rapid relief of exercise fatigue by soothing and regulating the human phototropic system, reducing vagal tone, and accelerating the excretion of metabolites; while PNF stretching relieved the exercise fatigue by providing physical and verbal communication to transfer the perception of fatigue, and by promoting the excretion of metabolites through muscle isometric contraction. 2. The effect of electric auto-massage therapy was marginally stronger than the commonly used PNF stretching exercise method.

Clinical efficacy of vibration stimulation therapy to relieve acute exercise fatigue.

BACKGROUND: Previous studies showed that local vibration stimulation therapy was effective in relieving fatigue, and the effects of different modes of vibration stimulation therapy were further investigated. OBJECTIVE: This study aimed to examine the effects of different vibration stimulation modes on relieving acute exercise fatigue based on the multiphysiological indicators such as heart rate variability (HRV), skin conductance level (SCL), and ratings of perceived exertion (RPE) subjective scale. METHODS: Sixty participants selected from the dragon boat team of the Shanghai University of Traditional Chinese Medicine were divided into acupoint stimulation group (20 participants), muscle stimulation group (20 participants), and control group (20 participants) by complete randomization. RESULTS: (1) RPE: both stimulation groups showed a significant increase compared to the control group. (2) Heart rate values: the difference between muscle stimulation group and control group was statistically significant; (3) SCL: the two stimulation groups had significantly higher and statistically significant differences in SCL (max) and SCL (mean) values compared to the control group; the muscle stimulation group had statistically significant differences in SCL (min) compared to the control group, and the acupoint stimulation group had statistically significant differences in SCL (v) compared to the control group; (4) HRV (hf): The difference between the acupoint stimulation group and the muscle stimulation group was statistically significant. CONCLUSION: (1) Both stimulation groups are part of vibration therapy, which can relieve sympathetic tension and regulate the vegetative nervous system's relaxation effect. (2) The meridian-vessel theory may be related to the acupoint stimulation group. The low-level visceral regulation centers in the spinal nerve segment region, where the acupoints are located, trigger changes in autonomic tone and enhance parasympathetic nerve activity to relieve acute motor fatigue. (3) The muscle stimulation group may be due to the 30-Hz vibration frequency's ability to raise muscle epidermal temperature, which increases blood flow and reflexively inhibits sympathetic excitation.

Response Surface Optimized Infrared-Assisted Extraction and UHPLC Determination of Flavonoid Types from Flos Sophorae.

Single-factor experiment and Box-Behnken design were applied to optimize the infrared-assisted extraction (IRAE) of rutin, quercetin, kaempferol, and isorhamnetin from Flos sophorae. Four factors (extract solvent, solid-liquid ration, extraction time, infrared power) affecting the extraction yield of flavonoids were studied. Under optimized conditions the extraction yield was 33.199 ± 0.24 mg/g, which substantially improved, compared with heating reflux extraction (HRE) and ultrasonic-assisted extraction (UAE), while extraction time was only 9 min. The eluents were rich in 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azobis (2-methyl-propionamidine) dihydrochloride radical scavenging potential (IC50 of DPPH: 53.44 ± 0.01 µg/mL, oxygen radical absorbance capacity (ORAC): 3785.83 ± 52 µmol/g) than the extracts obtained by HRE and UAE. In addition, an ultra-high performance liquid chromatography method was optimized for the identification and quantification of the tested flavonoids, and the method was validated based on its correlation coefficient (r), reproducibility (RSD, n = 5), and recovery values, which were 0.9994-0.9998, 0.74-1.83%, and 97.78-102.94%, respectively. These results confirmed that high extraction yield of flavonoids results in stronger antioxidant values and response surface methodology optimization of IRAE is a promising alternative to traditional extraction techniques for flavonoids from medicinal plants.

Application of near-infrared spectroscopy for the rapid quality assessment of Radix Paeoniae Rubra.

Near-infrared (NIR) spectroscopy with multivariate analysis was used to quantify gallic acid, catechin, albiflorin, and paeoniflorin in Radix Paeoniae Rubra, and the feasibility to classify the samples originating from different areas was investigated. A new high-performance liquid chromatography method was developed and validated to analyze gallic acid, catechin, albiflorin, and paeoniflorin in Radix Paeoniae Rubra as the reference. Partial least squares (PLS), principal component regression (PCR), and stepwise multivariate linear regression (SMLR) were performed to calibrate the regression model. Different data pretreatments such as derivatives (1st and 2nd), multiplicative scatter correction, standard normal variate, Savitzky-Golay filter, and Norris derivative filter were applied to remove the systematic errors. The performance of the model was evaluated according to the root mean square of calibration (RMSEC), root mean square error of prediction (RMSEP), root mean square error of cross-validation (RMSECV), and correlation coefficient (r). The results show that compared to PCR and SMLR, PLS had a lower RMSEC, RMSECV, and RMSEP and higher r for all the four analytes. PLS coupled with proper pretreatments showed good performance in both the fitting and predicting results. Furthermore, the original areas of Radix Paeoniae Rubra samples were partly distinguished by principal component analysis. This study shows that NIR with PLS is a reliable, inexpensive, and rapid tool for the quality assessment of Radix Paeoniae Rubra.

Community Physician-Guided Long-Term Domiciliary Oxygen Therapy Combined With Conventional Therapy in Stage IV COPD Patients.

PURPOSE: The aim of the study was to explore clinical effect of community physician-guided long-term domiciliary oxygen therapy (LTDOT) on patients with Stage IV chronic obstructive pulmonary disease (COPD). DESIGN: A retrospective study. METHODS: Fifty-four patients with Stage IV COPD were recruited and randomly divided into two groups (the LTDOT group and the control group). Patients in LTDOT group accepted additional oxygen therapy for more than 15 hours every day with continuous low flow (1-2 L/min) for 3 years. FINDINGS: PaO2 (O2 pressure), FEV1/FVC (forced vital capacity), and FEV1% (percentage of forced expiratory volume in 1 second) in the LTDOT group increased significantly after treatment. A significant decrease was observed on the BODE index in the LTDOT group (p < .05) but not in control group (p > .05). Frequencies and costs of hospitalization therapy and emergency medical services were markedly decreased after 3 years of LTDOT. CONCLUSION: Community physician-guided LTDOT can improve prognosis and reduce the costs for stage IV COPD patients. CLINICAL RELEVANCE: Rehabilitation nurses can be instrumental in helping patients with stage IV COPD learn principles of LTDOT.

[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.

An effective solution to discover synergistic drugs for anti-cerebral ischemia from traditional Chinese medicinal formulae.

Recently, the pharmaceutical industry has shifted to pursuing combination therapies that comprise more than one active ingredient. Interestingly, combination therapies have been used for more than 2500 years in traditional Chinese medicine (TCM). Understanding optimal proportions and synergistic mechanisms of multi-component drugs are critical for developing novel strategies to combat complex diseases. A new multi-objective optimization algorithm based on least angle regression-partial least squares was proposed to construct the predictive model to evaluate the synergistic effect of the three components of a novel combination drug Yi-qi-jie-du formula (YJ), which came from clinical TCM prescription for the treatment of encephalopathy. Optimal proportion of the three components, ginsenosides (G), berberine (B) and jasminoidin (J) was determined via particle swarm optimum. Furthermore, the combination mechanisms were interpreted using PLS VIP and principal components analysis. The results showed that YJ had optimal proportion 3(G): 2(B): 0.5(J), and it yielded synergy in the treatment of rats impaired by middle cerebral artery occlusion induced focal cerebral ischemia. YJ with optimal proportion had good pharmacological effects on acute ischemic stroke. The mechanisms study demonstrated that the combination of G, B and J could exhibit the strongest synergistic effect. J might play an indispensable role in the formula, especially when combined with B for the acute stage of stroke. All these data in this study suggested that in the treatment of acute ischemic stroke, besides restoring blood supply and protecting easily damaged cells in the area of the ischemic penumbra as early as possible, we should pay more attention to the removal of the toxic metabolites at the same time. Mathematical system modeling may be an essential tool for the analysis of the complex pharmacological effects of multi-component drug. The powerful mathematical analysis method could greatly improve the efficiency in finding new combination drug from TCM.

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).

By improving regional cortical blood flow, attenuating mitochondrial dysfunction and sequential apoptosis galangin acts as a potential neuroprotective agent after acute ischemic stroke.

Ischemic stroke is a devastating disease with a complex pathophysiology. Galangin is a natural flavonoid isolated from the rhizome of Alpina officinarum Hance, which has been widely used as an antioxidant agent. However, its effects against ischemic stroke have not been reported and its related neuroprotective mechanism has not really been explored. In this study, neurological behavior, cerebral infarct volumes and the improvement of the regional cortical blood flow (rCBF) were used to evaluate the therapeutic effect of galangin in rats impaired by middle cerebral artery occlusion (MCAO)-induced focal cerebral ischemia. Furthermore, the determination of mitochondrial function and Western blot of apoptosis-related proteins were performed to interpret the neuroprotective mechanism of galangin. The results showed that galangin alleviated the neurologic impairments, reduced cerebral infarct at 24 h after MCAO and exerted a protective effect on the mitochondria with decreased production of mitochondrial reactive oxygen species (ROS). These effects were consistent with improvements in the membrane potential level (Dym), membrane fluidity, and degree of mitochondrial swelling in a dose-dependent manner. Moreover, galangin significantly improved the reduced rCBF after MCAO. Western blot analysis revealed that galangin also inhibited apoptosis in a dose-dependent manner concomitant with the up-regulation of Bcl-2 expression, down-regulation of Bax expression and the Bax/Bcl-2 ratio, a reduction in cytochrome c release from the mitochondria to the cytosol, the reduced expression of activated caspase-3 and the cleavage of poly(ADP-ribose) polymerase (PARP). All these data in this study demonstrated that galangin might have therapeutic potential for ischemic stroke and play its protective role through the improvement in rCBF, mitochondrial protection and inhibiting caspase-dependent mitochondrial cell death pathway for the first time.

[Pharmacological research of wild and cultivated Atractylodes lancea].

OBJECTIVE: To explore the differences of the main pharmacological action in wild and cultivated Atractylodes lancea. METHOD: Study the effect of the 70% ethanol extracts from the wild and the culticated A. lancea on the mice with gastriculcer induced by absolute ethyl alcohol or HCl. Stdudy the effect on intestinal propulsive function in mice by measuring the length of intestine and distance of Indian ink. Observe anti-bacteria effect in vitro of the A. lancea by determining the minimum inhibitory concentrations. RESULT: The pharmacological action, which made a little difference in the sane productive places, and made significant differences in different productive places. Compared to Jurong, it has better actions in Luotian on small intestinal propulsive function and bacteriostatic action. There was not an obvious difference in protective effect on gastric ulcer induced by absolute ethyl alcohol and 0.6 mol x L(-1) HC1 between different locations. CONCLUSION: To the main effect, cultvated A. lancea had no significant difference on the whole but having better result in some respects, which can provide the pharmacodynamics evidence of cultivated A. lancea being used instead of the wild.

[Determination of 5-hydroxy-7-methoxyflavan in Daemonorops draco by HPLC].

OBJECTIVE: To establish a method for the determination of 5-hydroxy-7-methoxyflavan in Daemrnonorops draco by HPLC. METHOD: Dikma Diamonsil C15 column(4. 6 mm x 150 mm, 5 microm) was used. The mobile phase consisted of (60:40). The temperature of column was 30 degrees C. The flow rate was 1:0 mL x min(-1) and the detection wavelength was at 210 nm. RESULT: The calibration curve was in good linearity with in the range from 0.01 to 0.10 microg (r = 0.9999). The average recovery was 100.1% (n=6). CONCLUSION: The method is simple, rapid, as well as precise and reliable, and can be used for the determination of 5-hydroxy-7-methoxyflavan in D. draco.

[Advances in chemistry, molecular biology and pharmacological of Cangzhu].

We described chemical composition in Cangzhu in recent years, volatile oil is the important chemical composition, The beta-eudesmol, hinesol are active ingredient in volatile oil and there are 38 kind of glycosides. At the same time, we overview the applying of RAPD technology in atractylodes lancea. The results is that there are correlation in chemical composition, genetic differentiation and geographical distribution, there is some truth in bounded by a territorial division of the north-south Cangzhu, and genetic differentiation has been happened in atractylodes lancea to adapting the environment. We described advances of pharmacological in dampness spleen, cardiovascular system, genitourinary system, nervous system, and the results show that there are pharmacological activity in digestive system, cardiovascular system, genitourinary system of atractylodes lancea.

[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.

[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.

[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.