Telomere-to-telomere genome assembly of Oldenlandia diffusa.

We report the complete telomere-to-telomere genome assembly of Oldenlandia diffusa which renowned in traditional Chinese medicine, comprising 16 chromosomes and spanning 499.7 Mb. The assembly showcases 28 telomeres and minimal gaps, with a total of only five. Repeat sequences constitute 46.41% of the genome, and 49,701 potential protein-coding genes have been predicted. Compared with O. corymbosa, O. diffusa exhibits chromosome duplication and fusion events, diverging 20.34 million years ago. Additionally, a total of 11 clusters of terpene synthase have been identified. The comprehensive genome sequence, gene catalog, and terpene synthase clusters of O. diffusa detailed in this study will significantly contribute to advancing research in this species' genetic, genomic, and pharmacological aspects.

First report of powdery mildew caused by Podosphaera fusca on Coreopsis tinctoria in China.

Coreopsis tinctoria is an annual herb and commonly cultivated in gardens due to its attractive flowers, its capitula also have been used as a traditional medicine in China, Asia, North America and Europe (Shen et al. 2021). In June 2023, severe powdery mildew infection was observed on C. tinctoria in a hillside near headwork of the middle route of the South to North Water Diversion Project (32°40'55''N, 111°41'59''E). Abundant irregular white spots were found on adaxial surface of the leaves and tender stems. Approximately 75% of the observed C. tinctoria plants showed these signs and symptoms. Generative hyphae were thin-walled, smooth or almost so, and 5 to 9 µm wide. Conidiophores were unbranched, straight, 80.5 to 162.5 × 9.3 to 12.9 µm (n=25), and produced one to three immature conidia. Foot-cells of conidiophores were cylindrical, 38.5 to 62.3 µm (n=20) long. Conidia were ellipsoid to ovoid, 25.1 to 31.9 × 15.2 to 19.5 µm (n=30). The morphological characteristics of asexual structures corresponded to Podosphaera sp. (Braun and Cook 2012). For further identification, genomic DNA was extracted directly from the mycelia and conidia using Chelex 100 (Sigma Aldrich, Shanghai, China). The internal transcribed spacer (ITS) regions and 28S large subunit (LSU) of ribosomal DNA from the specimen (CT2302) were amplified using the primers ITS1/ITS4 (expected amplicon size 566 bp) (White et al. 1990) and NL1/NL4 (expected amplicon size 618 bp) (Baten et al. 2014), respectively. The sequences of ITS (GenBank accession no. OR649304) and LSU (GenBank accession no. OR649305) showed 99.63% and 100% identity values to the Podosphaera fusca isolate HMNWAFU-CF2012074 in the NCBI database (KR048109 for ITS and KR048178 for LSU), respectively. Phylogenetic analyses based on the combined ITS and LSU sequences using MEGA 7.0 software indicated that CT2302 formed a monophyletic clade together with isolates of P. fusca. Therefore, this fungus was identified as P. fusca based on the morphological and molecular characteristics. Pathogenicity tests were performed by gently pressing the infected leaves onto 15 young leaves of five healthy plants and three noninoculated plants were used as controls. All plants were maintained in a greenhouse (25℃ and 70% relative humidity). Powdery mildew symptoms similar to those of originally diseased plants were observed on all inoculated leaves after 12 days, whereas no symptoms were observed on the control leaves. Powdery mildew caused by P. fusca (previously Sphaerotheca fusca) on C. tinctoria has been reported in Russia, Poland, Korea, Romania and Ukraine (Cho and Shin 2004; Rusanov and Bulgakow 2008). To our knowledge, this is the first report of P. fusca on C. tinctoria in China. The identification of P. fusca as the causal agent on C. tinctoria is critical to the prevention and control of this disease in the future.

Exploring the in vitro anti-inflammatory activity of gross saponins of Tribulus terrestris L. fruit by using liquid chromatography-mass spectrometry-based cell metabolomics approach.

Our previous studies confirmed the efficacy of gross saponins of Tribulus terrestris L. fruit in treating cerebral ischemia. This study aimed to investigate the related mechanisms in vitro. The lipopolysaccharide-induced BV2 cells model was constructed and treated with gross saponins at different concentrations to explore its anti-inflammatory activity. The cell metabolite changes were tracked by liquid chromatography-mass spectrometry (LC-MS)-based metabolomics, and the metabolic biomarkers and related metabolic pathways were analyzed. Molecular biochemistry analysis was further used to verify the relevant inflammatory pathways. The results showed that the saponins reduced nitric oxide release and the secretion of tumor necrosis factor-alpha, interleukin-1ß, and interleukin-6 from lipopolysaccharide-induced BV2 cells. Metabolic perturbations occurred in lipopolysaccharide-treated BV2 cells, which could be reversed by drug treatment via mainly regulating glycerophospholipid metabolism, tryptophan metabolism, purine metabolism pathways, etc. The western blot analysis demonstrated that saponin could suppress the activation of the inflammatory-related signaling pathway. The present study explored the in vitro anti-inflammatory mechanism of gross saponins of Tribulus terrestris L. fruit using an LC-MS-based cell metabolomics approach, which confirms the great potential of LC-MS for drug efficacy evaluation and can be applied in other herbal medicine-related analyses.

Rapid Discrimination of Panax quinquefolium and Panax ginseng Using the Proofman-Duplex-LMTIA Technique.

This study aims to establish a rapid identification method based on the Proofman-LMTIA technique for distinguishing between Panax quinquefolium and Panax ginseng. By targeting specific 18S rDNA sequences, suitable primers and Proofman probes labeled FAM or JOE were designed for LMTIA. Initially, single-species-primer Proofman-LMTIA assays were performed separately for each ginseng type to optimize reaction temperature, assess sensitivity and specificity, and determine the detection limit. Subsequently, both sets of primers and their corresponding probes were combined in the same reaction system to further optimize reaction conditions, evaluate sensitivity, and assess stability. Finally, the developed Proofman-duplex-LMTIA technique was employed to detect P. quinquefolium and P. ginseng slices available in the market. Single-plex Proofman-LMTIA assays revealed that the optimal reaction temperature for both P. quinquefolium and P. ginseng was 62 °C. The sensitivity was as low as 1 pg/µL, with a detection limit of 0.1%, and both showed excellent specificity. The optimal temperature for Proofman-duplex-LMTIA assays was 58 °C. This method could simultaneously identify P. quinquefolium and P. ginseng. Testing 6 samples of P. ginseng and 11 samples of P. quinquefolium from the market resulted in a 100% positive rate for all samples. This study successfully established a rapid, simple, sensitive, and specific Proofman-duplex-LMTIA identification method for P. quinquefolium and P. ginseng. It provides an effective means for quality control of P. quinquefolium, P. ginseng, and related products.

Investigating the in vivo effect of Tribulus terrestris extract in middle cerebral artery occlusion rats using LC-MS-based metabolomics combined with molecular docking.

Tribulus terrestris L. fruit (TT) is a traditional Chinese herbal medicine used to treat ischemic stroke (IS). This study aimed to investigate the protective effect of TT extract, named TT15, on middle cerebral artery occlusion (MCAO) rats using metabolomics and molecular docking and find the targets of action and the material basis of TT15 against IS. The results of the infarct volume and neurological defect scores confirmed the efficacy of TT15. Serum metabolomics analysis using LC-MS revealed that model group animals experienced a variety of metabolic disturbances when compared to the sham group. TT15 can restore the MCAO-induced serum metabolite changes by modulating multiple metabolic pathways. Six enzymes were highlighted by the metabolite-reaction-enzyme-gene (M-R-E-G) network analysis, which might be the possible targets for the TT15 against IS. Molecular docking analysis was applied to show the binding affinities between active compounds and these enzymes. The representative docking mode with the lowest binding energy between three compounds and phospholipase A 2 (PLA2) and peroxidase (POD) was displayed by the ribbon binding map. This study profiles the metabolic changes in MCAO-induced IS and investigates the efficacy and the corresponding mechanism of TT15 in the treatment of IS.

Insights into the health status of the general population living near an electroplating industry zone: metal elevations and renal impairment.

A cross-sectional study was conducted in 2016 in Zhejiang Province, China, to evaluate the body burdens of metals and metalloids associated with renal dysfunction in populations living near electroplating industries. We recruited 236 subjects and performed physical examinations, determined the blood and urinary levels of arsenic (As), cadmium (Cd), chromium (Cr), manganese (Mn), nickel (Ni), lead (Pb), antimony (Sb), and selenium (Se) by an inductively coupled plasma mass spectrometer (ICP-MS), and measured three renal impairment biomarkers, namely nacetyl-ß-D-glucosaminidase (NAG), retinol-binding protein (RBP), and ß2-microglobulin (BMG). The proportion of abnormal nasal symptoms in the exposure group (10.1%) was much higher than in the control group (0; p < 0.05). The blood and urinary levels of As, Cd, and Se in the exposure group were significantly higher than those in the control group (p < 0.05). The blood levels of Mn and Pb, as well as the urinary levels of Cr and Ni, were significantly higher in the exposure group than in the control group (p < 0.05). The exposure group demonstrated higher levels of NAG, RBP, and BMG than the control group (0.51 vs. 0.14 mg/g creatinine, 12.79 vs. 9.26 IU/g creatinine, and 1.39 vs. 0.78 mg/g creatinine, respectively; p < 0.05). Urinary BMG was positively correlated with urinary Cd levels (r = 0.223, p < 0.05), while urinary RBP was correlated with blood Cd levels (r = 0.151, p < 0.05) and urinary Cd, Cr, Ni, and Se levels (r = 0.220, 0.303, 0.162, and 0.306, respectively; p < 0.05). In conclusion, our study indicated that a population living in the vicinity of electroplating industries had high body burdens of certain metals and metalloids associated with non-negligible renal dysfunction.

The Effect of Azithromycin Combined with Palm Massage on Pulmonary Function of Children with Mycoplasma Pneumonia.

Objective: The study aimed to determine the effect of azithromycin combined with palm massage on the pulmonary function of children suffering from mycoplasma pneumonia, as well as associated complications. Methods: A total of 60 cases of children suffering from mycoplasma pneumonia admitted to our hospital from May 2018 to May 2020 were selected as study subjects and assigned to either the observation group or the control group. Lung function indices and changes in the levels of inflammatory markers were measured before and after treatment in both groups, and the results were compared to assess treatment efficacy. Results: There was no statistically significant difference in lung function indices between the two groups before treatment (P > 0.05), while there was a statistically significant difference between the two groups after treatment (P < 0.05), with the observation group outperforming the control group. Similarly, there was no statistically significant difference in the levels of such inflammatory markers as IL-6 and TNF-α between the two groups before treatment (P > 0.05). After treatment, there was a decrease in the levels of inflammatory markers in both groups, but a much larger decrease was seen in the observation group (P < 0.05) than in the control group. The total treatment efficacy in the observation group was 93.3%, which was significantly higher than that of 70.0% in the control group (P < 0.05). Conclusion: The application of palm massage combined with azithromycin in the treatment of children suffering from mycoplasma pneumonia produces outstanding results in terms of relieving symptoms and improving pulmonary function.

Ribosomal DNA copy number associated with blood metal levels in school-age children: A follow-up study on a municipal waste incinerator in Zhejiang, China.

To evaluate the body burdens of heavy metals and explore the impact of environmental metal exposure on ribosomal DNA (rDNA) or mitochondrial DNA (mtDNA) copy number (CN) variation in school-age children living near a municipal waste incinerator (MWI), we conducted a follow-up study in 2019. A total of 146 sixth-grade children from a primary school located 1.2 km away from the MWI were recruited for our study. Metals, including vanadium (V), chromium (Cr), manganese (Mn), cobalt (Co), nickel (Ni), copper (Cu), zinc (Zn), arsenic (As), selenium (Se), cadmium (Cd), stannum (Sn), stibium (Sb), thallium (Tl), and lead (Pb), were determined by an inductively coupled plasma mass spectrometer method. Real-time qPCR was used to measure the rDNA and mtDNA CN. The blood metal levels followed this order: Zn > Cu > Se > Pb > Mn > Sb > As > Ni > Cd > Co > Cr > Sn > V > Tl. Blood Cr level was significantly correlated with 18 S, 2.5 S, and 45 S CN (ß = -0.25, -0.22, -0.26, p < 0.05); Ni was correlated with 5 S (ß = -0.36, p < 0.01); Cu was correlated with 28 S, 18 S, and 5.8 S (ß = -0.24, -0.24, -0.23, p < 0.05); while Zn was correlated with 18 S, 5.8 S, and 45 S (ß = -0.28, -0.32, -0.26, p < 0.05). In conclusion, school-age children living near the MWI had lower blood metal levels compared to children recruited in 2013, while rDNA CN loss was found to be correlated to several heavy metals in these children.

Stachydrine is effective and selective against blast phase chronic myeloid leukaemia through inhibition of multiple receptor tyrosine kinases.

CONTEXT: Resistance to BCR-ABL tyrosine kinase inhibitor (TKI) is the cause of treatment failure in blast phase chronic myeloid leukaemia (BP-CML). Agents that act synergistically with BCR-ABL TKI are required to improve response. OBJECTIVE: This work investigated the effects of stachydrine in CML. MATERIALS AND METHODS: CML cells were treated with control or stachydrine at 20, 40 and 80 µM. Proliferation and apoptosis were examined after 72 h treatment. Combination studies were performed in four groups: control, TKI, stachydrine and the combination of stachydrine and TKI. Immunoblotting analysis was performed in CML cells after 24 h treatment. RESULTS: Stachydrine inhibited K562 (IC50 61 µM), KCL22 (IC50 141 µM), LAMA84 (IC50 86 µM), Ba/F3 T315I (IC50 26 µM), Ba/F3 WT (IC50 22 µM) and KU812 (IC50 35 µM) proliferation, and induced apoptosis in these CML cell lines. Stachydrine significantly induced apoptosis, inhibited colony formation and self-renewal in BP-CML CD34+ cells. The combination index of stachydrine and TKI combination was <1. Compared to TKI alone, the combination of stachydrine and TKI significantly induced more apoptosis and decreased colony formation in BP-CML CD34+ cells. Stachydrine decreased phosphorylation levels of multiple receptor tyrosine kinases in CML cells. DISCUSSION AND CONCLUSIONS: Our study is the first to demonstrate (1) the anticancer activity of stachydrine on primary patient cancer cells; (2) the inhibitory effects of stachydrine on cancer stem cells; (3) the synergism between stachydrine and other anticancer drugs.

Discussion on "Correct and logical causal inference for binary and time-to-event outcomes in randomized controlled trials" by Yi Liu, Bushi Wang, Miao Yang, Jianan Hui, Heng Xu, Siyoen Kil, and Jason C. Hsu.

In their paper, Liu et al. (2020) pointed out illogical discrepancies between subgroup and overall causal effects for some efficacy measures, in particular the odds and hazard ratios. As the authors show, the culprit is subgroups having prognostic effects within treatment arms. In response to their provocative findings, we found that the odds and hazard ratios are logic respecting when the subgroups are purely predictive, that is, the distribution of the potential outcome for the control treatment is homogeneous across subgroups. We also found that when we redefined the odds and hazards ratio causal estimands in terms of the joint distribution of the potential outcomes, the discrepancies are resolved under specific models in which the potential outcomes are conditionally independent. In response to other discussion points in the paper, we also provide remarks on association versus causation, confounding, statistical computing software, and dichotomania.

[Expression of Runt-related Transcription Factor 3 in Human Colon Cancer Cell Line HCT-116 Resistant to 5-Fluorouracil and the Mechanism of Drug Resistance].

Objective To establish a human colon cancer cell line HCT-116/5-FU resistant to 5-fluorouracil(5-FU)and explore the relationship between runt-related transcription factor 3(RUNX3)and drug resistance of colorectal cancer.Methods The human colon cancer cell line HCT-116/5-FU with resistance to 5-FU was established by low concentration gradient increment combined with high-dose intermittent shock.CCK-8 method was used to determine the half maximal inhibitory concentration(IC50)of 5-FU on the parent line HCT-116 and drug-resistant line HCT-116/5-FU.The cell growth curve was established for the calculation of population doubling time(TD).The mRNA levels and protein levels of RUNX3,P-glycoprotein(P-gp),multidrug resistance-associated protein 1(MRP1),and lung resistance-related protein(LRP)in HCT-116 and HCT-116/5-FU cells were determined by qRT-PCR and Western blotting,respectively.The RUNX3 expression in HCT-116 cells was knocked down by siRNA technique,and the cells were divided into RUNX3 knockdown groups(si-RUNX3-1 group and si-RUNX3-2 group)and negative control group(si-NC group).The knockdown efficiency was verified by qRT-PCR at the mRNA level and Western blotting at the protein level.The IC50 in si-RUNX3 groups and si-NC group was determined with CCK-8 method,and the expression of P-gp,MRP1,and LRP in the two groups was detected by Western blotting.Results A stable human colon cancer drug-resistant cell line HCT-116/5-FU was successfully constructed.HCT-116/5-FU showed the TD 1.38 times as long as that of HCT-116(P=0.002)and changed morphology.The mRNA level of RUNX3 in HCT-116/5-FU cells was significantly lower than that in HCT-116 cells(P=0.048),and those of P-gp(P=0.008),MRP1(P=0.001),and LRP(P=0.001)showed the opposite trend.The protein level of RUNX3 in HCT-116/5-FU cells was significantly lower than that in HCT-116(P<0.001),and those of P-gp,MRP1,and LRP presented the opposite trend(all P<0.001).The HCT-116 cell model with low expression of RUNX3 was successfully established.The mRNA level of RUNX3 had no significant difference between si-RUNX3-1 group and si-NC group(P=0.064),while the level in si-RUNX3-2 group was significantly lower than that in si-NC group(P=0.034).The protein levels of RUNX3 in si-RUNX3-1 group and si-RUNX3-2 group were lower than that in si-NC group(both P<0.001).The results demonstrated higher knocking efficiency in si-RUNX3-2 group,which was thus selected to complete the follow-up test.The IC50 of si-RUNX3 group was significantly higher than that of si-NC group(P<0.001),which indicated that the down-regulated expression of RUNX3 could reduce the sensitivity of HCT-116 cells to 5-FU.The relative protein levels of P-gp,MRP1,and LRP in si-RUNX3 group were significantly higher than those in si-NC group(all P<0.001).Conclusion The down-regulation of RUNX3 expression can reduce the sensitivity of HCT-116 cells to 5-FU,which is considered to be related to the up-regulated expression of P-gp,MRP1,and LRP.

[A preliminary report on the variation of respiratory heart rate during sleep in normal subjects and patients with chronic diseases without sleep apnea].

Objective: The new theory of holistic integrative physiology and medicine, which describes the integrative regulation of respiratory, circulatory and metabolic systems in human body, generates the hypothesis of that breath is the origin of variability of circulatory parameters. We investigated the origin of heart rate variability by analyzing relationship between the breath and heart rate variability (HRV) during sleep. Methods: This retrospective study analyzed 8 normal subjects (NS) and 10 patients of chronic diseases without sleep apnea (CDs-no-SA). After signed the informed consent form, they performed cardiopulmonary exercise testing (CPET) in Fuwai Hospital and monitored polysomnography (PSG) and electrocardiogram (ECG) during sleep since 2014. We dominantly analyzed the correlation between the respiratory cycle during sleep and the heart rate variability cycle of the ECG R-R interval. The HRV cycle included the HR increase from the lowest to the highest and decrease from the highest to the lowest point. The number of HRV (HRV-n), average HRV time and other parameters were calculated. The breath cycle included complete inhalation and subsequent exhalation. The number of breath (B-n), average breath time and other breath parameters were analyzed and calculated. We analyzed each person's relationship between breath and HRV; and the similarities and differences between the NS and CDs-no-SA groups. Independent sample t test was used for statistical analysis, with P<0.05. Results: CPET core parameter such as Peak VO2 (83.8±8.9)% in NS were significantly higher than that (70.1±14.9)% in patients of chronic diseases without sleep apnea (P<0.05), but there was no difference between their AHI (1.7±1.3) in NS and AHI (2.9±1.2) in CDs-no-SA (P>0.05). The B-n and the HRV-n (6581.63±1411.90 vs 6638.38±1459.46), the average B time and the average HRV time (4.19±0.57)s vs (4.16±0.62)s in NS were similar without significant difference (P>0.05). The comparison of the numbers in CDs-no-SA were the number (7354.50±1443.50 vs 7291.20±1399.31) and the average times ((4.20±0.69)s vs (4.23±0.68)s) of B and HRV were similar without significant difference (P>0.05). The ratios of B-n/HRV-n in NS and CDs-no-SA were (0.993±0.027 vs 1.008±0.024) and both were close to 1 and similar without significant difference (P>0.05). The average magnitude of HRV in NS ((5.74±3.21) bpm) was significantly higher than that in CDs-no-SA ((2.88±1.44) bpm) (P<0.05). Conclusion: Regardless of the functional status of NS and CDs-no-SA, there is a similar consistency between B and HRV. The origin of initiating factors of HRV is the respiration.

[Preliminary analysis of the influence of breathing on heart rate variability in chronically ill patients with sleep apnea].

Objective: Based on the hypothesis that respiration causes variability of circulatory indicators proposed by the holistic integrated physiology and medicine theory, the correlation between respiration and heart rate variability during sleep in chronically ill patients with abnormal sleep breathing is analyzed. Methods: Eleven chronically ill patients with abnormal sleep breathing and apnea-hypopnea index (AHI) ≥15 times/hr are recruited. After signing the informed consent, they completed the standardized symptomatic restrictive extreme exercise cardiopulmonary exercise testing (CPET) and sleep breathing monitoring Calculate and analyze the rules of respiratory nasal airflow and ECG RR interval heart rate variability during the oscillatory breathing (OB) phase and the normal steady breathing phase of the patient during sleep, and use the independent sample t test to compare with normal people and no sleep breathing abnormalities in the same period in this laboratory. Of patients with chronic diseases are more similar and different. Results: The peak oxygen uptake and anaerobic threshold (AT) of CPET in chronic patients with abnormal sleep apnea were (70.8±13.6)% Pred and (71.2±6.1)% Pred; 5 cases of CPET had exercise induced oscillatory breathing (EIOB), 6 An example is unstable breathing, which indicates that the overall functional status is lower than normal. In this group of patients with chronic diseases, AHI (28.8±10.0) beats/h, the ratio of the total time of abnormal sleep breathing to the total time of sleep (0.38±0.25); the length of the OB cycle (51.1±14.4)s. The ratio (Bn/HRV-B-n) of the number of breathing cycles in the normal and steady breathing period to the number of heart rate variability cycles in this group of patients with chronic diseases is 1.00±0.04, and the CV (SD of HRV-B-M/x) is (0.33 ±0.11), blood oxygen saturation (SpO2) did not decrease significantly, the average amplitude of heart rate variability (HRV-B-M) of each respiratory cycle rhythm was (2.64±1.59) bpm, although it was lower than normal people (P<0.05) , But it was similar to chronic patients without sleep apnea (P>0.05). In this group of patients with chronic diseases, the ratio of the number of respiratory cycles to the number of heart rate variability cycles (OB-Bn/OB-HRV-B-n) during OB is (1.22±0.18), and the average amplitude of heart rate variability for each respiratory cycle rhythm in OB (OB -HRV-B-M) is (3.56±1.57)bpm and its variability (OB-CV = SD of OB-HRV-B-M/x) is (0.59±0.28), the average amplitude of heart rate variability in each OB cycle rhythm (OB-HRV-OB-M) is (13.75±4.25)bpm, SpO2 decreases significantly during hypoventilation during OB, and the average decrease in SpO2 during OB (OB-SpO2-OB-M) is (4.79±1.39)%. The OB-Bn/OB-HRV-B-n ratio, OB-HRV-OB-M and OB-SpO2-OB-M in the OB period are all significantly higher than the corresponding indicators in the normal stable breathing period Large (P<0.01). Although OB-HRV-B-M has no statistically significant difference compared with HRV-B-M in normal stable breathing period (P>0.05), its variability OB-CV is significantly increased (P<0.01). Conclusion: The heart rate variability of chronic patients with abnormal sleep breathing in the OB phase is greater than that of the normal stable breathing period. When the breathing pattern changes, the heart rate variability also changes significantly. The number of breathing cycles in the stable breathing period is equal to the number of heart rate variability cycles.The ratio is the same as that of normal people and chronically ill patients without sleep apnea, confirming that heart rate variability is respiratory origin; and the reduction of heart rate variability relative to the respiratory cycle during OB is directly caused by hypopnea or apnea at this time, and heart rate variability is also breathing source.

[The new theory CPET guides the overall plan of individualized precision exercise to effectively improve the overall functional status of "frailty"].

Objective: Under the guidance of the new theory of holistic integrated physiology and medicine, the effect of individualized accurate exercise program on the overall functional state was studied according to cardiopulmonary exercise testing (CPET). Methods: Li xx, female, 31 years old, has a fast heart rate since childhood (90~100 bpm), usually feel cold, especially in autumn and winter, and general health good. CPET was performed after signing the informed consent form at Fuwai Hospital in September 2019. Peak oxygen uptake, anaerobic threshold (AT), and peak cardiac output were (69~72)% pred, respectively, and the oxygen uptake ventilation efficiency and carbon dioxide exhaust ventilation efficiency were basically normal (96~100)% pred. The resting heart rate was fast, the blood pressure was low, the blood pressure response was weak during exercise, and the heart rate was mainly increased. The holistic integrated physiology medical theory pointed out that she was in weak health and heart weakness was the main manifestation. CPET was used to guide individualized precise exercise intensity titration, combine continuous beat-by-beat blood pressure, ECG, pulse and blood glucose dynamic monitoring to formulate an holisticplan of individualized quantitative exercise .Reexamine CPET after 8 weeks' strengthening management. Results: After 8 weeks of intensive holistic management, the limbs were warm and the cold symptoms disappeared. Re-examination of CPET peak oxygen uptake, AT and peak cardiac output were (90~98)% pred, which increased by (30~36)% respectively, and the holistic weak functional status was significantly improved; basically normal oxygen uptake ventilation efficiency and carbon dioxide exhaust ventilation efficiency also increased by (10~37)% respectively; resting heart rate and blood pressure basically returned to normal, and blood pressure and heart rate response during exercise were normal. Continuous ambulatory blood glucose monitoring indicated that the average blood glucose level decreased slightly and became more stable. Repeated measurement results of continuous ECG and beat-to-beat blood pressure also indicated a decrease in heart rate and an increase in blood pressure during rest, exercise and during sleep, and radial pulse wave. The amplitude of the dicrotic wave increases and becomes more pronounced. Conclusion: The new theoretical system to guide CPET to formulate an holistic plan for individualized precision exercise can safely and effectively enhance myocardial contractility, increase stroke volume, increase blood pressure, lower heart rate, stabilize and slightly lower blood glucose, and improve holistic functional status.

Comparative Analysis of the Effect of Low-Frequency Repeated Transcranial Magnetic Stimulation and Extracorporeal Shock Wave on Improving the Spasm of Flexor after Stroke.

Poststroke spasticity (PSS) patients with muscle spasticity are effectively relieved by low-frequency repetitive transcranial magnetic stimulation (rTMS) or extracorporeal shock wave treatment (ESWT). However, there are relatively few reports about the difference in the efficacy of rTMS and ESWT for PSS. In this study, we examined and recorded the levels of UE motor section of the Fugl-Meyer Motor Assessment Scale (FMA-UE), myoelectric signal time-domain range integral values (iEMG), Modified Ashworth Scale (MAS), and Modified Barthel Index (MBI) before and after treatment to observe the differences in treatment effects between rTMS and ESWT in patients with PSS. 66 patients with PSS were enrolled in the study and signed an informed consent form, and the study was approved by the Ethics Committee of the First Hospital of Soochow University (2019008). The patients were divided into rTMS group, ESWT group, and regular group according to the random number table method, and there were 22 patients in each group. The rTMS group and ESWT group were treated with rTMS and ESWT on the basis of conventional treatment in the regular group, 5 times a week, and the total treatment time was 4 weeks. The results of the study showed that iEMG, MAS, FMA-UE, and MBI scores in the rTMS, ESWT, and regular groups were significantly ameliorated after treatment compared with those before treatment. The efficacy of the ESWT group was significantly better than in the regular group and slightly better than in the rTMS group, as shown by the iEMG, MAS, FMA-UE, and MBI scores, and the iEMG score of the ESWT group was significantly better than the rTMS group, while there were no significant differences in other indexes. The FMA-UE and MBI scores in the rTMS group were significantly better than those in the regular group after treatment in the rTMS group; however, the comparison between iEMG and MAS scores was not statistically significant. It can be seen that both rTMS and ESWT can alleviate upper limb flexor spasm, improve upper limb motor function, and improve activities of daily living in patients with PSS. Among them, ESWT has better antispasmodic effect and better short-term treatment effect.

Enzyme-Powered Liquid Metal Nanobots Endowed with Multiple Biomedical Functions.

Catalytically powered micro/nanobots (MNBs) can perform active movement by harnessing energy from in situ chemical reactions and show tremendous potential in biomedical applications. However, the development of imageable MNBs that are driven by bioavailable fuels and possess multiple therapeutic functions remains challenging. To resolve such issues, we herein propose enzyme (urease) powered liquid metal (LM) nanobots that are naturally of multiple therapeutic functions and imaging signals. The main body of the nanobot is composed of a biocompatible LM nanoparticle encapsulated by polydopamine (PDA). Urease enzyme needed for the powering and desired drug molecules, e.g., cefixime trihydrate antibiotic, are grafted on external surfaces of the PDA shell. Such a chemical composition endows the nanobots with dual-mode ultrasonic (US) and photoacoustic (PA) imaging signals and favorable photothermal effect. These LM nanobots exhibit positive chemotaxis and therefore can be collectively guided along a concentration gradient of urea for targeted transportation. When exposed to NIR light, the LM nanobots would deform and complete the function change from active drug carriers to photothermal reagents, to achieve synergetic antibacterial treatment by both photothermal and chemotherapeutic effects. The US and PA properties of the LM nanoparticle can be used to not only track and monitor the active movement of the nanobots in a microfluidic vessel model but also visualize their dynamics in the bladder of a living mouse in vivo. To conclude, the LM nanobots demonstrated herein represent a proof-of-concept therapeutic nanosystem with multiple biomedical functionalities, providing a feasible tool for preclinical studies and clinical trials of MNB-based imaging-guided therapy.

Multi-omics analysis of brain tissue metabolome and proteome reveals the protective effect of gross saponins of Tribulus terrestris L. fruit against ischemic stroke in rat.

ETHNOPHARMACOLOGICAL RELEVANCE: Gross Saponins of Tribulus terrestris L. Fruit (GSTTF) has been reported to have a protective effect against ischemic stroke, but the related mechanism is complex and still not fully investigated. AIM OF THE STUDY: The combination of metabolomics and proteomics approach was applied to reveal the mechanisms of GSTTF in treating ischemic stroke. MATERIALS AND METHODS: The metabolite and protein changes in brain tissue were analyzed by the LC-MS-based untargeted metabolomics method and tandem mass tags (TMT)-based quantitative proteomics technology. The multivariate statistical analysis and protein-protein interaction (PPI) analysis were conducted to screen out the biomarkers, and their related pathway was further investigated by the joint pathway analysis. RESULTS: A total of 110 metabolites and 359 differential proteins, which were mainly associated with complement and coagulation cascades, sphingolipid metabolism, glycerophospholipid metabolism, glutathione metabolism, and platelet activation, etc. were screened out from the rat brain tissue. The PPI network exhibited that the protein F2, Fga, Fgb, Fgg, Plg, and C3, which are greatly involved in the complement and coagulation cascades, have a relatively high connectivity degree, indicating their importance in the process of middle cerebral artery occlusion (MCAO). The GSTTF exerted a protective effect against MCAO via modulating multiple proteins on this pathway. Moreover, F2 played a key role during the protective process and worth to be further investigated due to it has been reported as one of the therapeutic targets of ischemic stroke. CONCLUSION: The present study could improve the understanding of the potential therapeutic mechanism of GSTTF against ischemic stroke.

First report of powdery mildew of crape jasmine (Tabernaemontana divaricata) caused by Erysiphe elevata in China.

Crape jasmine (Tabernaemontana divaricata) is a popular flowering shrub widely grown in southern China. Its leaves and roots are used in Chinese traditional medicine. In December, 2019, powdery mildew symptoms were observed on five crape jasmine shrubs on the campus of Shenzhen Polytechnic (22°35'N; 113°56'E), in Guangdong province. Approximately 45% of leaves were infected. Symptoms initially appeared as circular to irregular white patches on the leaf petiole, and subsequently coalesced to develop into abundant hyphal growth on both sides of the leaves, which soon wilted. Hyphae were septate, branched, with simple kidney-shaped to moderately lobed appressoria. Conidia formed singly, ellipsoid-ovoid to subcylindrical, 27-37 × 14-20 µm (mean 32±2.5 × 17±1.6 µm), with a length/width ratio varying from 1.3 to 2.4. Conidiophores were erect, unbranched, consisted of two cells, 60 to 84 µm long (mean 73±4 µm), and with straight to severely kinked cylindrical foot-cells at the base, 29-35 × 3-7 µm (mean 32±3 × 6±2 µm). Chasmothecia were not observed on sampled plants. These morphological characteristics were typical to the conidial stage of the genus Erysiphe (Braun and Cook, 2012). For molecular identification, genomic DNA was extracted from conidia washed from infected leaves and using Fungal DNA Kit (Omega Bio-tek Inc., Guangzhou, China). Semi-nested PCR amplification of the internal transcribed spacer (ITS) region of rDNA was conducted by using primer sets P3 (Kusaba et al., 1995)/ITS5 and ITS5/ITS4 (White et al., 1990) for the first and second reactions, respectively. BLASTn analysis of the obtained 719 bp sequence (GenBank Accession No. MT802112) showed 99.7% identity with those of E. elevata (KY660910, MH985631, MK253282). On the basis of morphological and molecular analyses, the fungus was identified as Erysiphe elevata. To confirm pathogenicity, infected leaves were gently pressed onto healthy leaves of three healthy plants in separate pots, and three noninoculated plants were used as controls. All plants were maintained in a greenhouse at 25 ℃, and relative humidity of 50 to 65%. After 11 days, similar disease symptoms were observed on the inoculated plants while no symptoms developed on control plants. The fungus observed on the inoculated shrubs was identical morphologically to that o the original infected leaves. E. elevata is a common powdery mildew species infecting Catalpa spp. (Cook et al., 2006), Plumeria rubra (Wu et al., 2019; Yeh et al., 2019) and Eucalyptus camaldulensis (Meeboon and Takamatsu, 2017). However, no powdery mildew were found on P. rubra nearby. To our knowledge, this is the first report of this fungus infecting T. divaricata.

Inhibition of mutant Kras and p53-driven pancreatic carcinogenesis by atorvastatin: Mainly via targeting of the farnesylated DNAJA1 in chaperoning mutant p53.

Recent studies have indicated that using statins to inhibit the mevalonate pathway induces mutant p53 degradation by impairing the interaction of mutant p53 with DnaJ subfamily A member 1 (DNAJA1). However, the role of the C-terminus of DNAJA1 with a CAAX box for farnesylation in the binding, folding, and translocation of client proteins such as mutant p53 is not known. In the present study, we used a genetically engineered mouse model of pancreatic carcinoma and showed that atorvastatin significantly increased animal survival and inhibited pancreatic carcinogenesis. There was a dramatic decrease in mutant p53 protein accumulation in the pancreatic acini, pancreas intraepithelial neoplasia lesions, and adenocarcinoma. Supplementation with farnesyl pyrophosphate, a substrate for protein farnesylation, rescued atorvastatin-induced mutant p53 degradation in pancreatic cancer cells. Tipifarnib, a farnesyltransferase inhibitor, mirrored atorvastatin's effects on mutant p53, degraded mutant p53 in a dose-dependent manner, and converted farnesylated DNAJA1 into unfarnesylated DNAJA1. Farnesyltransferase gene knockdown also significantly promoted mutant p53 degradation. Coimmunoprecipitation either by an anti-DNAJA1 or p53 antibody confirmed the direct interaction of mutant p53 and DNAJA1 and higher doses of atorvastatin treatments converted more farnesylated DNAJA1 into unfarnesylated DNAJA1 with much less mutant p53 pulled down by DNAJA1. Strikingly, C394S mutant DNAJA1, in which the cysteine of the CAAX box was mutated to serine, was no longer able to be farnesylated and lost the ability to maintain mutant p53 stabilization. Our results show that farnesylated DNAJA1 is a crucial chaperone in maintaining mutant p53 stabilization and targeting farnesylated DNAJA1 by atorvastatin will be critical for inhibiting p53 mutant cancer.

Inhibition of Pancreatic Carcinoma Growth Through Enhancing ω-3 Epoxy Polyunsaturated Fatty Acid Profile by Inhibition of Soluble Epoxide Hydrolase.

BACKGROUND/AIM: Cytochrome P450 epoxygenase is a major enzyme involved in the metabolism of ω-3 polyunsaturated fatty acids (PUFAs) to produce biologically active ω-3 epoxy fatty acids (ω-3 epoxides). In general, all epoxy PUFAs including ω-3 epoxides are quickly metabolized/inactivated by soluble epoxide hydrolase (sEH) to form diol products. The aims of this study were to determine the effect and mechanism of fat-1 transgene, and ω-3 PUFA combined with sEH gene knockout or inhibitor on inhibiting pancreatic cancer and the related mechanisms involved. MATERIALS AND METHODS: PK03-mutant KrasG12D murine pancreatic carcinoma cells were inoculated into mouse models including fat-1, sEH-/- and C57BL/6J mice. The mice were fed with AIN-76A diet with or without ω-3 PUFA supplementation or treated with sEH inhibitor. In addition to tumor growth (tumor size and weight), cell proliferation, mutant Kras-mediated signaling, inflammatory reaction and angiogenesis were analyzed immunohisto-chemically and by western blot assay. ω-3 PUFA metabolism, particularly focusing on ω-3 epoxy fatty acids (ω-3 epoxides), was measured using a liquid chromatography with tandem mass spectrometry (LC-MS/MS) approach. RESULTS: Significant decreases of weight and size of the PK03 pancreatic carcinoma were observed in the fat-1 transgenic mice treated with sEH inhibitor compared to those of C57BL/6J control mice fed with AIN-76A diet (weight: 0.28±0.04 g vs. 0.58±0.06 g; size: 187.0±17.5 mm3 vs. 519.3±60.6 mm3). In a separate experiment, sEH-/- mice fed ω-3 PUFA supplement and C57BL/6J mice treated with sEH inhibitor and fed ω-3 PUFA supplement exhibited a significant reduction in the weight and size of the pancreatic carcinoma compared to C57BL/6J control mice (weight: 0.26±.26 g and 0.39±.39 g vs. 0.69±0.11 g, respectively; size: 274.2±36.2 mm3 and 296.4±99.8 mm3 vs. 612.6±117.8 mm3, respectively). Moreover, compared to the pancreatic tumors in C57BL/6J control mice, the tumors in fat-1 transgenic mice treated with sEH inhibitor showed a significant less inflammatory cell infiltrate (62.6±9.2/HPF (high power field) vs. 8.0±1.2/HPF), tumor cell proliferation (48.5±1.7% vs. 16.5±1.6%), and angiogenesis (micro-vessel density (MVD): 35.0±1.0 vs. 11.1±0.5) immunohistochemically, as well as significantly increased caspase-3 labeled apoptosis (0.44±0.06% vs. 0.69±0.06%, respectively). Using western blot approach, significant inhibition of mutant Kras-activated signals including phosphorylated Serine/threonine kinases (cRAF), Mitogen-activated protein kinase kinase (MEK), and extracellular signal-regulated kinase (ERK) were identified in pancreatic carcinoma of fat-1 transgenic mice treated with sEH inhibitor. Eicosanoic acid metabolic profiling of the serum specimens detected a significant increase of the ratios of epoxides to dihydroxy fatty acid (DiHDPE) for docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), and epoxides/dihydroxy octadecenoic acid (DiHOME) for arachidonic acid (ARA) and linoleic acid (LA), as well as a significant increase of epoxy metabolites of DHA, EPA, ARA and LA in fat-1 transgenic mice treated with a sEH inhibitor. CONCLUSION: ω-3 epoxy products from ω-3 PUFA metabolism play a crucial role in inhibiting pancreatic cancer growth, and use of ω-3 PUFAs combined with sEH inhibition is a strategy with high potential for pancreatic cancer treatment and prevention.