Characterization of CsUGT73AC15 as a Multifunctional Glycosyltransferase Impacting Flavonol Triglycoside Biosynthesis in Tea Plants.

Flavonol glycosides, contributing to the health benefits and distinctive flavors of tea (Camellia sinensis), accumulate predominantly as diglycosides and triglycosides in tea leaves. However, the UDP-glycosyltransferases (UGTs) mediating flavonol multiglycosylation remain largely uncharacterized. In this study, we employed an integrated proteomic and metabolomic strategy to identify and characterize key UGTs involved in flavonol triglycoside biosynthesis. The recombinant rCsUGT75AJ1 exhibited flavonoid 4'-O-glucosyltransferase activity, while rCsUGT75L72 preferentially catalyzed 3-OH glucosylation. Notably, rCsUGT73AC15 displayed substrate promiscuity and regioselectivity, enabling glucosylation of rutin at multiple sites and kaempferol 3-O-rutinoside (K3R) at the 7-OH position. Kinetic analysis revealed rCsUGT73AC15's high affinity for rutin (Km = 9.64 µM). Across cultivars, CsUGT73AC15 expression inversely correlated with rutin levels. Moreover, transient CsUGT73AC15 silencing increased rutin and K3R accumulation while decreasing their respective triglycosides in tea plants. This study offers new mechanistic insights into the key roles of UGTs in regulating flavonol triglycosylation in tea plants.

Genome-Wide Investigation of Oxidosqualene Cyclase Genes Deciphers the Genetic Basis of Triterpene Biosynthesis in Tea Plants.

Triterpenoids from Camellia species comprise a diverse class of bioactive compounds with great therapeutic potential. However, triterpene biosynthesis in tea plants (Camellia sinensis) remains elusive. Here, we identified eight putative 2,3-oxidosqualene cyclase (OSC) genes (CsOSC1-8) from the tea genome and characterized the functions of five through heterologous expression in yeast and tobacco and transient overexpression in tea plants. CsOSC1 was found to be a ß-amyrin synthase, whereas CsOSC4, 5, and 6 exhibited multifunctional α-amyrin synthase activity. Molecular docking and site-directed mutagenesis showed that the CsOSC6M259T/W260L double mutant yielded >40% lupeol, while the CsOSC1 W259L single mutant alone was sufficient for lupeol production. The V732F mutation in CsOSC5 altered product formation from friedelin to taraxasterol and ψ-taraxasterol. The L254 M mutation in the cycloartenol synthase CsOSC8 enhanced the catalytic activity. Our findings shed light on the molecular basis governing triterpene diversity in tea plants and offer potential avenues for OSC engineering.

Metabolomics integrated with machine learning to discriminate the geographic origin of Rougui Wuyi rock tea.

The geographic origin of agri-food products contributes greatly to their quality and market value. Here, we developed a robust method combining metabolomics and machine learning (ML) to authenticate the geographic origin of Wuyi rock tea, a premium oolong tea. The volatiles of 333 tea samples (174 from the core region and 159 from the non-core region) were profiled using gas chromatography time-of-flight mass spectrometry and a series of ML algorithms were tested. Wuyi rock tea from the two regions featured distinct aroma profiles. Multilayer Perceptron achieved the best performance with an average accuracy of 92.7% on the training data using 176 volatile features. The model was benchmarked with two independent test sets, showing over 90% accuracy. Gradient Boosting algorithm yielded the best accuracy (89.6%) when using only 30 volatile features. The proposed methodology holds great promise for its broader applications in identifying the geographic origins of other valuable agri-food products.

Clinical characteristics of re-hospitalized COVID-19 patients with recurrent positive SARS-CoV-2 RNA: a retrospective study.

At present, it has been noticed that some patients recovered from COVID-19 present a recurrent positive RNA test of SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) after being discharged from hospitals. The purpose of the current study was to characterize the clinical features of re-hospitalized patients with recurrent SARS-CoV-2 positive results. From January 12 to April 1 of 2020, our retrospective study was conducted in China. The exposure history, baseline data, laboratory findings, therapeutic schedule, and clinical endpoints of the patients were collected. All the patients were followed until April 10, 2020. Among all COVID-19 patients included in the current study, there were 14 re-hospitalized patients due to recurrent positive tests of SARS-CoV-2 RNA. Fever (11 [78.6%]), cough (10 [71.4%]), and fatigue (7 [50.0%]) were the most common symptoms on the patient's first admission, and less symptoms were found on their second admission. The average duration from the onset of symptoms to admission to hospital was found to be 8.4 days for the first admission and 2.6 days for the second admission (P = 0.002). The average time from the detection of RNA (+) to hospitalization was 1.9 days for the first admission and 2.6 days for the second admission (P = 0.479), and the average time from RNA (+) to RNA (-) was 11.1 days for the first admission and 6.3 days for the second admission (P = 0.030). Moreover, the total time in hospital was 18.6 days for the first admission and 8.0 days for the second admission (P = 0.000). It may be necessary to increase the isolation observation time and RT-PCR tests should be timely performed on multiple samples as soon as possible.

Case report of a 28-year-old man with aortic dissection and pulmonary shadow due to granulomatosis with polyangiitis.

BACKGROUND: Granulomatosis with polyangiitis (GPA) is characterised by the main violation of the upper and lower respiratory tract and kidney. GPA is considered a systemic vasculitis of medium-sized and small blood vessels where aortic involvement is extremely rare. CASE PRESENTATION: A 28-year-old male was admitted to the hospital due to 4 h of chest pain. Computed tomography scan of the aorta showed a thickened aortic wall, pulmonary lesions, bilateral pleural effusion and pericardial effusion. The aortic dissection should be considered. An emergency operation was performed on the patient. Surgical biopsies obtained from the aortic wall showed destructive changes, visible necrosis, granulation tissue hyperplasia and a large number of acute and chronic inflammatory cells. Nearly a year later, the patient was re-examined for significant pulmonary lesions. His laboratory studies were significantly positive for anti-neutrophilic antibody directed against proteinase 3. Finally, the diagnosis of GPA was obviously established. CONCLUSIONS: Although GPA rarely involves the aorta, we did not ignore the fact that GPA may involve large blood vessels. In addition, GPA should be included in the systemic vasculitis that can give rise to aortitis and even aortic dissection.

[Analysis of pressure-time and flow-time curve in ventilator].

OBJECTIVE: To better understand the significance of the pressure-time curve and flow-time curve from the perspective of PB840 ventilator working principle. METHODS: (1) Mechanical principle: flow supply valves (air valve and oxygen valve) and exhalation valve in PB840 ventilator were controlled to achieve the ventilation target (volume or pressure) by the central processing unit according to the monitoring data from pressure sensors (P1 at the supply side, P2 at the exhalation side) and flow sensors (Q1 at the air side, Q2 at the oxygen side, Q3 at the exhalation side). (2) The essence of curve: each point means a value of pressure or flow at a certain time measured by the sensors or calculated by the system. (3) The respiratory process could be divided into inspiratory part, expiratory part, and the connection part from expiratory to inspiratory. The air running state and the respiratory mechanics relationship at the three parts could be inferred according to the form of curves. RESULTS: (1) Inspiratory process: at volume-controlled and constant flow ventilation: there should be a relationship "Pc-Pa = XR" between alveolar pressure (Pa) and circuit pressure (Pc) according to Ohm law. So, the Pc curve (pressure-time curve) could indirectly reflect the Pa curve with the flow (X) and resistance (R) being constant. At pressure-set ventilation: it is the goal of ventilator to maintain the Pc at the target level. So, the stability of the target pressure line in pressure-time curve reflects the matching ability of the flow supply valves and the exhalation valve. (2) Expiratory process: it could be divided into pre-expiratory [without basic flow (Ba) or bias flow (Bi)] and post-expiratory (with Ba or Bi), where Ba or Bi is equal to "Q1+Q2". So, the mathematical function are "X(t) = Q3t" in pre-part, and "X(t) = Q3t-(Q1t+Q2t)" in post-part. The relationship between pressure and flow at peak expiratory flow point: it could be found that there is an obvious time span and area formation under the curve from 0 to peak point (Fpeak) after stretching the abscissa axis of flow-time curve. It means that some gas have been discharged from the lung when it arrives at the peak point. So, the alveolar pressure should be lower than the platform pressure at the point (Pplat). The circuit pressure is significantly higher than positive end expiratory pressure (PEEP) at the point in the stretching axis diagram. So, it means that the formula "RE = (Pplat-PEEP)/Fpeak" to calculate the expiratory resistance (E) is unreasonable in the angle of Ohm law. (3) The process from exhalation to inspiratory: according to the difference of the starting point of the conversion, it could be divided into two cases: one is that the inspiratory started from the ending of exhalation. Here, the inhaling starting point is lying in the abscissa axis. The other is that the inspiratory started before the ending of exhalation (with endogenous positive end expiratory pressure). Here, the starting point is lying below the abscissa axis, and the slope of the following curve is obviously larger than the slope of natural expiratory curve. According to the difference of results from the starting point to the end of the inhalation triggering effort, it could be divided into two cases: one is that it reach the trigger point. Here, the expiratory curve extends upward from or below the horizontal axis until an effective air supply is triggered. The other is that it could not reach the trigger point. Here, the expiratory curve extends upward from or below the horizontal axis, but then runs downward (meaning exhaling). CONCLUSIONS: It is helpful to analyze the ventilation state, ventilation failure, and the causes of man-machine confrontation with understanding the ventilation principle and the air route map of the ventilator.

A negative regulation loop of long noncoding RNA HOTAIR and p53 in non-small-cell lung cancer.

Non-small-cell lung cancer (NSCLC) is one of the leading causes of cancer-related death worldwide, and the 5-year survival rate is still low despite advances in diagnosis and therapeutics. A long noncoding RNA (lncRNA) HOX antisense intergenic RNA (HOTAIR) has been revealed to play important roles in NSCLC carcinogenesis but the detailed mechanisms are still unclear. In the current study, we aimed to investigate the regulation between the lncRNA HOTAIR and p53 in the NSCLC patient samples and cell lines. Our results showed that HOTAIR expression was significantly higher in the cancer tissues than that in the adjacent normal tissue, and was negatively correlated with p53 functionality rather than expression. When p53 was overexpressed in A549 cells, the lncRNA HOTAIR expression was downregulated, and the cell proliferation rate and cell invasion capacity decreased as a consequence. We identified two binding sites of p53 on the promoter region of HOTAIR, where the p53 protein would bind to and suppress the HOTAIR mRNA transcription. Inversely, overexpression of lncRNA HOTAIR inhibited the expression of p53 in A549 cells. Mechanistic studies revealed that HOTAIR modified the promoter of p53 and enhanced histone H3 lysine 27 trimethylation (H3K27me3). These studies identified a specific negative regulation loop of lncRNA HOTAIR and p53 in NSCLC cells, which revealed a new understanding of tumorigenesis in p53 dysfunction NSCLC cells.

[Variation and clinical value of endothelial glycocalyx in the patients with septic shock].

OBJECTIVE: To explore the variation and clinical value of the degradation of endothelial glycocalyx in the patients with septic shock. METHODS: A prospective case control study was conducted. Patients of 18 years or older diagnosed with septic shock and admitted to Department of Critical Care Medicine of Affiliated Hospital of Binzhou Medical University from June 2014 to May 2015 were enrolled. The levels of degradation products, including hyaluronic acid (HA) and heparin sulfate (HS), at 0, 6, 12, 24, 48 hours were determined, while 20 healthy people were enrolled and served as controls. The changes of HA and HS were analyzed in the patients with septic shock. The differences of HA and HS between survival group and death group after 28 days were also analyzed. The relationships between HA, HS and tumor necrosis factor-α (TNF-α), sequential organ failure assessment (SOFA) score, arterial blood lactate (Lac), platelet, albumin were analyzed by Pearson correlation analysis. The receiver-operating characteristic (ROC) curve was plotted to assess the prognostic value of HA and HS for patients with septic shock. RESULTS: Thirty-one patients diagnosed as septic shock were enrolled, among whom 17 patients died after 28 days, with a mortality of 54.8%. The levels of HA and HS in patients with septic shock were increased significantly as compared with those of health control group, peaked at 48 hours, and the levels of HA and HS at 48 hours were significantly higher than those at 0 hour [HA (µg/L): 119.47±32.44 vs. 94.84±23.63, HS (µg/L): 72.83±19.03 vs. 58.83±16.63, both P < 0.05]. The levels of HA and HS at 0 hour and 48 hours in death group were significantly higher than those of the survival group [HA (µg/L): 130.42±27.67 vs. 93.29±29.80, 105.14±19.18 vs. 70.82±13.24; HS (µg/L): 67.23±25.01 vs. 39.23±14.58, 79.74±19.84 vs. 56.17±14.53, all P < 0.05]. The levels of HA and HS in patients with septic shock were remarkably positively correlated with the levels of TNF-α, SOFA score, Lac, and platelet, but were remarkably negatively correlated with albumin levels (r value of HA was 0.595, 0.462, 0.545, 0.466, -0.534, respectively; r value of HS was 0.607, 0.468, 0.563, 0.547, -0.455, respectively; all P < 0.05). It was demonstrated by ROC curves that the areas under ROC curve (AUC) of HA and HS at 0 hour and 48 hours for predicting the prognosis of patients with septic shock were 0.881, 0.940 and 0.833, 0.821, respectively, the sensitivities of HA and HS were 87.5%, 100.0% and 83.3%, 81.3%, respectively, and the specificities of HA and HS were 82.6%, 78.3% and 91.3%, 78.3%, respectively. CONCLUSIONS: The concentrations of degradation products generated by endothelial glycocalyx in the blood of the patients with septic shock are remarkably increased. The elevated levels of the degradation products are closely associated with the severity of septic shock, microcirculation disturbance, and the levels of inflammatory factors.

Effects of ultrasound-guided radial artery catheterization: an updated meta-analysis.

BACKGROUND: Previous meta-analyses have shown that ultrasound guidance is an effective technique for radial artery catheterization. However, these reports neglected to include several non-English language studies. Therefore, an updated meta-analysis including more eligible studies was performed to assess the effectiveness of ultrasound-guided radial artery catheterization. METHODS: Eligible studies were identified by systematically searching PubMed, EMBASE, Wanfang, and China National Knowledge Infrastructure literature databases. The outcome measure was the rate of first-attempt success. Two investigators identified the randomized controlled trials (RCTs) for inclusion and independently extracted data from these RCTs. The quality of the included studies was evaluated using the Jadad score. The relative risk (RR) for dichotomous outcomes and the 95% confidence intervals (CIs) were calculated and pooled using a random-effects model. RESULTS: Eleven RCTs involving 803 patients met the inclusion criteria. Ultrasound-guided radial artery catheterization was generally associated with a 47% improvement, as compared with the palpation technique, in terms of the rate of first-attempt success (RR, 1.47; 95% CI, 1.22-1.76; P < .0001). Specifically, the ultrasound-guided technique significantly improved the rate of first-attempt success for adult (RR, 1.39; 95% CI, 1.13-1.72; P = .002) and pediatric (RR, 1.68; 95% CI, 1.15-2.47; P = .008) patients. CONCLUSIONS: Adult and pediatric patients benefited from ultrasound-guided radial artery catheterization in terms of the rate of first-attempt success. Given the potential bias and significant heterogeneity of the available data in the present study, further investigation is required to confirm the present findings and to identify other effects of the ultrasound-guided technique.

[The significance of lactic acid in early diagnosis and goal-directed therapy of septic shock patients].

OBJECTIVE: To investigate the application of lactic acid in early diagnosis and goal-directed therapy of septic shock, and to provide reference for the early clinical diagnosis and treatment of septic shock. METHODS: A prospective observational study was conducted, in which patients satisfied with the criteria of septic shock diagnosis were enrolled. The patients were randomly divided into two groups. The lactic group was defined using blood lactic acid concentration < 2 mmol/L as treatment guide target. Control group was defined according to the traditional diagnostic criteria of shock which systolic blood pressure was less than 90 mmHg (1 mmHg= 0.133 kPa) or systolic blood pressure value fell > 40 mmHg baseline or oliguria ( < 0.5 ml. kg-1.h-1) et al traditional septic shock diagnosis criteria and bundle treatment was performed. Organ dysfunction index, the sequential organ failure score (SOFA), acute physiology and chronic health evaluation score II ( APACHE II) score, the time of mechanical ventilation, the time of stay in the intensive care unit ( ICU), and the 7-and 28-day mortality were recorded. RESULTS: There were 26 and 31 septic shock patients in lactic group and control group respectively. Organ dysfunction index had been improved in different degrees after treatment compared with that before treatment. Creatinine ( Cr) at 48 hours after treatment in lactic group was significantly lower than that in control group (µmol/L: 94.48 ± 6.68 vs. 107.44 ± 10.35, P < 0.05), and there was no statistical difference in other indexes. The SOFA score of lactic group at 24 hours and 48 hours after treatment was lower than that of control group (9.27 ± 4.62 vs. 9.79 ± 3.80, t=2.103, P=0.040; 8.54 ± 5.53 vs. 9.70 ± 4.30, t=2.302, P=0.023). APACHE II score of two group after treatment were lower than that before treatment, and lactic group decreased more obviously compared with control group ( 14.25 ± 5.29 vs. 20.00 ± 9.74, t=2.298, P=0.026; 13.60 ± 6.18 vs 18.15 ± 6.62, t=2.653, P=0.011). The time of stay in the ICU and the time of mechanical ventilation of lactic group were shorter that those of control group, but there was no statistical difference [ICU time (days): 8.95 ± 5.19 vs. 9.45 ± 6.18, t=0.605, P=0.652; mechanical ventilation time (hours): 101.15 ± 11.50 vs. 110.63 ±13.26, t= 0.631, P=0.564]. There was no statistical difference regarding 7-day mortality of lactic group was lower than that of control group [15.38% (4/26) vs. 16.13% ( 5/31), Χ2=0.000, P=1.000]. The 28-day mortality of lactic group was lower than that of control group [26.92 % (7/26) vs. 54.84% (17/31). Χ2=4.520, P=0.033]. CONCLUSION: By blood lactic acid monitoring, inadequacy of organization perfusion i.e. septic shock can be found in the early stage, thus early intervention can be performed and improve resuscitation result and reduce the mortality of septic shock patients. Blood lactic acid ≥ 4 mmol/L can be used as one of the criteria for the diagnosis of septic shock, and 6-hour blood lactic acid < 2 mmol/L as a goal to guide shock treatment with obvious prognosis improvement.

miR-137 impairs the proliferative and migratory capacity of human non-small cell lung cancer cells by targeting paxillin.

Human lung cancer is the leading cause of cancer motility worldwide, with nearly 1.4 million deaths each year, among which non-small cell lung cancer (NSCLC) accounts for almost 85% of this disease. The discovery of microRNAs (miRNAs) provides a new avenue for NSCLC diagnostic and treatment regiments. Currently, a large number of miRNAs have been reported to be associated with the progression of NSCLC, among which serum miR-137 has been examined to be down-regulated in NSCLC patients. However, the function of miR-137 on NSCLC cells migration and invasion and the relative mechanisms were less known. Here, we found that ectopic expression of miR-137 could inhibit cell proliferation, induce cell apoptosis, and suppress cell migration and invasion in NSCLC cell line A549. Moreover, we found that paxillin (PXN) was a target gene of miR-137 in NSCLC cells and restored expression of PXN abolished the miR-137-mediated suppression of cell migration and invasion. Taken together, our results showed that miR-137 acted as a tumor suppressor in NSCLC by targeting PXN, and it may provide novel diagnostic and therapeutic options for human NSCLC clinical operation in future.

[Comparison of the effects of BiPAP ventilation combined with lung recruitment maneuvers and low tidal volume A/C ventilation in patients with acute respiratory distress syndrome].

OBJECTIVE: To compare the effects of BiPAP ventilation combined with lung recruitment maneuvers (LRM) with low tidal volume A/C ventilation in patients with acute respiratory distress syndrome (ARDS). METHODS: A prospective, randomized comparison of BiPAP mechanical ventilation combined with lung recruitment maneuvers (test group) with low tidal volume A/C ventilation (control group) was conducted in 28 patients with ARDS. FiO2/PaO2 ratio, respiratory system compliance (Cs), central venous pressure (CVP), duration of ventilation support were recorded at 0 h, 48 h and 72 h separately. The ventilation associated lung injury and mortality at 28 d were also recorded. RESULTS: The FiO2/PaO2 ratio were (298+/-16) and (309+/-16) cm H2O, Cs were (38.4+/-2.2) and (42.0+/-1.3) ml/cm H2O, CVP were (13.8+/-0.8) and (11.6+/-0.7) cm H2O in the test group at 48 h and 72 h separately. In the control group, FiO2/PaO2 ratio were (212+/-12) and (246+/-17) cm H2O, Cs were (29.5+/-1.3) and (29.0+/-1.0) ml/cm H2O, CVP were 18.6+/-1.1 and (16.8+/-1.0) cm H2O. The results were better in the test group as compared with the control group (t=10.03-29.68, all P<0.01). The duration of ventilation support in the test group was shorter than the control group [(14+/-3) d vs (19+/-3) d, t=4.80, P<0.01]. The mortality in 28 d and ventilation associated lung injury were similar in the two groups. CONCLUSION: The results show that combination of LRM with BiPAP mode ventilation, as compared with the control group, contributes to improved FiO2/PaO2 ratio, pulmonary compliance, stable hemodynamic and shorter duration of ventilation support in patients with ARDS.

[Efficacy of goal-directed therapy in the treatment of septic shock].

OBJECTIVE: To investigate the efficacy and effect on outcome of goal-directed therapy in patients with septic shock compared with conventional therapy. METHODS: Sixteen patients with septic shock were randomly assigned to receive goal-directed therapy, with central venous pressure (CVP) 8-12 mm Hg (1 mm Hg=0.133 kPa), mean arterial pressure (MAP) >or=65 mm Hg, venous oxygen saturation (SvO(2))>0.70 (superior vena cava saturation), and urine output >or=0.5 ml/min as therapeutic goals. Another 17 patients received conventional therapy as controls. The arterial oxygen saturation (SaO(2)), SvO(2), MAP, CVP, heart stroke volume cardiac index (CI), serum lactate, volume of fluid, amount of vasopressors, the numbers of organ injured and patients who needed continuity blood purification (CBP) and/or ventilation were recorded serially for 6-48 hours, and they were compared between the two groups. The mortality of the patients in two groups on 7 days and 14 days were also recorded. RESULTS: There were no significant differences between the groups with respect to base-line characteristics. During the interval from 24 to 48 hours, the patients assigned to goal-directed therapy had a significantly higher in SaO(2), SvO(2), MAP, CVP, CI (P<0.05 or P<0.01), a lower lactate concentration (P<0.01), significantly more fluid during 6-24 hours and less vasopressors (both P<0.01). Seven and 14 days in-hospital mortality were lower in goal-directed therapy group as compared with the control group(P<0.05). CONCLUSION: The efficacy of goal-directed therapy in patients with septic shock is significantly better than conventional therapy in ameliorating outcome of shock and can be easily used in intensive care unit (ICU).