A quadratic isothermal amplification fluorescent biosensor without intermediate purification for ultrasensitive detection of circulating tumor DNA.

Circulating tumor DNA (ctDNA) is an auspicious tumor biomarker released into the bloodstream by tumor cells, offering abundant information concerning cancer genes. It plays a crucial role in the early diagnosis of cancer. However, due to extremely low levels in body fluids, achieving a simple, sensitive, and highly specific detection of ctDNA remains challenging. Here, we constructed a purification-free fluorescence biosensor based on quadratic amplification of ctDNA by combining nicking enzyme mediated amplification (NEMA) and catalytic hairpin assembly (CHA) reactions. After double isothermal amplification, this biosensor achieved an impressive signal amplification of nearly 107-fold, enabling it to detect ctDNA with ultra-sensitivity. And the detection limit of this biosensor is as low as 2 aM. In addition, we explored the influence of human serum on the performance of the biosensor and found that it showed favorable sensitivity in the presence of serum. This biosensor eliminates the need for an intermediate purification step, resulting in enhanced sensitivity and convenience. Thus, our purification-free fluorescent biosensor exhibits ultra-high sensitivity when compared to other biosensors and has the potential to serve as an effective diagnostic tool for early detection of cancer.

The Monitoring and Cell Imaging of Fe3+ Using a Chromone-Based Fluorescence Probe.

A new structurally simple fluorescent CP probe based on chromone was designed and synthesized, and its structure was fully characterized using various analytical techniques. The CP probe displays a high selectivity and sensitivity for sensing Fe3+ with a "turn-off" fluorescence response over other metal ions in a DMSO/H2O (4:1, v/v) solution. The experiment results show that the CP probe is stable over a wide pH range of 2.0-12.0. The detection limit for Fe3+ was calculated to be 0.044 µmol•L-1. The molar ratio method indicated that the binding mode between the CP probe and Fe3+ is a 1:1 complex formation. HR-MS and density functional theory (DFT) calculations were also performed to further confirm the recognition mechanism. Both fluorescence imaging experiments and the MTT assay demonstrated that the CP probe was suitable for detecting intracellular Fe3+ and no significant cytotoxicity in living cells.

Chinese Medicine Formula Huashibaidu Granule Early Treatment for Mild COVID-19 Patients: An Unblinded, Cluster-Randomized Clinical Trial.

Background: Previous research suggested that Chinese Medicine (CM) Formula Huashibaidu granule might shorten the disease course in coronavirus disease 2019 (COVID-19) patients. This research aimed to investigate the early treatment effect of Huashibaidu granule in well-managed patients with mild COVID-19. Methods: An unblinded cluster-randomized clinical trial was conducted at the Dongxihu FangCang hospital. Two cabins were randomly allocated to a CM or control group, with 204 mild COVID-19 participants in each cabin. All participants received conventional treatment over a 7 day period, while the ones in CM group were additionally given Huashibaidu granule 10 g twice daily. Participants were followed up to their clinical endpoint. The primary outcome was worsening symptoms before the clinical endpoint. The secondary outcomes were cure and discharge before the clinical endpoint and alleviation of composite symptoms after the 7 days of treatment. Results: All 408 participants were followed up to their clinical endpoint and included in statistical analysis. Baseline characteristics were comparable between the two groups (P > 0.05). The number of worsening patients in the CM group was 5 (2.5%), and that in the control group was 16 (7.8%) with a significant difference between groups (P = 0.014). Eight foreseeable mild adverse events occurred without statistical difference between groups (P = 0.151). Conclusion: Seven days of early treatment with Huashibaidu granule reduced the likelihood of worsening symptoms in patients with mild COVID-19. Our study supports Huashibaidu granule as an active option for early treatment of mild COVID-19 in similar well-managed medical environments. Clinical Trial Registration:www.chictr.org.cn/showproj.aspx?proj=49408, identifier: ChiCTR2000029763.

Combination of Hua Shi Bai Du granule (Q-14) and standard care in the treatment of patients with coronavirus disease 2019 (COVID-19): A single-center, open-label, randomized controlled trial.

OBJECTIVE: To evaluate the efficacy and safety of Hua Shi Bai Du Granule (Q-14) plus standard care compared with standard care alone in adults with coronavirus disease (COVID-19). STUDY DESIGN: A single-center, open-label, randomized controlled trial. SETTING: Wuhan Jinyintan Hospital, Wuhan, China, February 27 to March 27, 2020. PARTICIPANTS: A total of 204 patients with laboratory-confirmed COVID-19 were randomized into the treatment group and control group, consisting of 102 patients in each group. INTERVENTIONS: In the treatment group, Q-14 was administered at 10 g (granules) twice daily for 14 days, plus standard care. In the control group, patients were provided standard care alone for 14 days. MAIN OUTCOME MEASURE: The primary outcome was the conversion time for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral assay. Adverse events were analyzed in the safety population. RESULTS: Among the 204 patients, 195 were analyzed according to the intention-to-treat principle. A total of 149 patients (71 vs. 78 in the treatment and control groups, respectively) tested negative via the SARS-CoV-2 viral assay. There was no statistical significance in the conversion time between the treatment group and control group (Full analysis set: Median [interquartile range]: 10.00 [9.00-11.00] vs. 10.00 [9.00-11.00]; Mean rank: 67.92 vs. 81.44; P = 0.051). The recovery time for fever was shorter in the treatment group than in the control group. The disappearance rate of symptoms like cough, fatigue, and chest discomfort was significantly higher in the treatment group. In chest computed tomography (CT) examinations, the overall evaluation of chest CT examination after treatment compared with baseline showed that more patients improved in the treatment group. There were no significant differences in the other outcomes. CONCLUSION: The combination of Q-14 and standard care for COVID-19 was useful for the improvement of symptoms (such as fever, cough, fatigue, and chest discomfort), but did not result in a significantly higher probability of negative conversion in the SARS-CoV-2 viral assay. No serious adverse events were observed. TRIAL REGISTRATION: ChiCTR2000030288.

Efficacy and safety assessment of severe COVID-19 patients with Chinese medicine: A retrospective case series study at early stage of the COVID-19 epidemic in Wuhan, China.

ETHNOPHARMACOLOGICAL RELEVANCE: The coronavirus disease 2019 (COVID-19) has formed a global pandemic since late 2019. Benefitting from the application experience of Chinese Medicine (CM) for influenza and SARS, CM has been used to save patients at the early stage of COVID-19 outbreak in China. AIM OF THE STUDY: In order to evaluate the efficacy and safety of CM, and compare with Western Medicine (WM) for COVID-19, we conducted a retrospective case series study based on the patients in Wuhan Jinyintan Hospital, Wuhan, China. METHODS: The inclusion and exclusion criteria of data extraction were set for this retrospective study. All patients who were admitted by the Wuhan Jinyintan Hospital between January 17th and February 25th 2020 were considered. In addition, patients enrolled met the severe defined by the guidelines released by the National Health Commission of the People's Republic of China. In these cases included in the study, CM or WM treatment was selected according to the wishes of the patients at the beginning of hospitalization. The patients in CM group were treated with Huashi Baidu granule (137 g po, bid) combined with the injections of Xiyanping (100 mg iv, bid), Xuebijing (100 ml iv, bid) and Shenmai (60 ml iv, qd) according to the syndrome of epidemic toxin blocking the lung in the theory of Traditional Chinese Medicine. The WM group received antiviral therapy (including abidor capsule 0.2 g po, tid; Lopinavir-Ritonavir tablets, 500 mg po, bid), antibiotics (such as cefoperazone 2 g iv, bid; moxifloxacin hydrochloride tablets, 0.4 g po, qd) or corticosteroid therapy (such as methylprednisolone succinate sodium 40 mg iv, qd; prednisone, 30 mg po, qd). In addition, patients in both groups received routine supportive treatment, including oxygen inhalation, symptomatic therapy, and/or human intravenous immunoglobulin, and/or serum albumin, and treatment for underlying diseases. The clinical outcomes were evaluated based on changes related with clinical manifestations, computer tomography (CT) scan images, and laboratory examinations before and after the treatment. RESULTS: 55 severe COVID-19 patients, with 23 in CM group and 32 in WM group, were included for analyzed. There was no case of death, being transferred to ICU, or receiving invasive mechanical ventilation in two groups during hospitalization. The median time of SARS-CoV-2 RNA clearance in CM and WM group were 12 days and 15.5 days respectively, the ratio of nucleic acid negative conversion of CM group at different follow-up time points was significantly higher than that of WM group (HR: 2.281, P = 0.018). Further, the chest CT imaging showed more widely lung lesion opacity absorbed in the CM group. The high sensitivity C-reactive protein and serum ferritin decreased significantly in the CM group (P<0.05). There was no significant difference in adverse events in terms of liver function and renal function between the two groups. CONCLUSION: Based on this retrospective analysis from Wuhan Jinyintan Hospital, CM has better effects in SARS-CoV-2 RNA clearance, promoting lung lesion opacity absorbed and reducing inflammation in severe COVID-19 patients, which is effective and safe therapy for treating severe COVID-19 and reducing mortality.

The Impact of Depression and Anxiety on Chronic Obstructive Pulmonary Disease Acute Exacerbations: A prospective cohort study.

BACKGROUND: To explore the impact of anxiety and depression in chronic obstructive pulmonary disease(COPD) patients on the risk of acute exacerbation. METHODS: 600 COPD patients were recruited and followed in Beijing, China. The demographic data, medical history, smoking history, therapy, assessments for anxiety and depression were completed by our physicians by face-to-face interview. Then the acute exacerbation events during the past one year was derived from their case record. RESULTS: 504 people (295 men and 209 women) were thus included in the final analyses.The patients with anxiety scored more than 50 showed higher risk of acute exacerbation(54.1%) than those who scoring less than 50(39.8%). Similarly, the patients with depression scored more than 53 showed higher risk of acute exacerbation(52.1%) than those who scoring less than 53(40.4%). Eventually, Anxiety and/or depression will increase the risk of acute exacerbation in chronic obstructive pulmonary disease patients(AECOPD) (adjusted OR = 1.60, 95%CI: 1.10-2.31), after adjusting the influence of family history, duration of disease, BMI index and other factors. Besides, the prevalence of acute exacerbation in patients with anxiety and/or depression was higher than those without acute exacerbation (P<0.05). LIMITATIONS: The frequency of AECOPD was limited to self-reported data, recall bias should be reconsidered, and it also may underestimate the association between anxiety and/or depression and AECOPD. Anxiety and depression were assessed by questionnaires rather than by a clinical diagnosis. CONCLUSION: Anxiety and/or depression in stable COPD patients were significantly associated with a higher risk of acute exacerbation. Mental health care should be paid more attention, to decrease the risk of acute exacerbation in COPD patients.

Efficacy and safety of Chinese herbal medicine versus Lopinavir-Ritonavir in adult patients with coronavirus disease 2019: A non-randomized controlled trial.

BACKGROUND: Treatments for coronavirus disease 2019 (COVID-19) are limited by suboptimal efficacy. METHODS: From January 30, 2020 to March 23, 2020, we conducted a non-randomised controlled trial, in which all adult patients with laboratory-confirmed COVID-19 were assigned to three groups non-randomly and given supportive treatments: Group A, Lopinavir-Ritonavir; Group B, Huashi Baidu Formula (a Chinese medicineformula made by the China Academy of Chinese Medical Sciences to treat COVID-19, which is now in the clinical trial period) and Lopinavir-Ritonavir; and Group C, Huashi Baidu Formula. The use of antibiotics, antiviruses, and corticosteroids was permitted in Group A and B. Traditional Chinese medicine injections were permitted in Group C. The primary outcomes were clinical remission time (interval from admission to the first time the patient tested negatively for novel coronavirus or an obvious improvement was observed from chest CT) and clinical remission rate (number of patients whose clinical time was within 16 days/total number of patients). RESULTS: A total of 60 adult patients with COVID-19 were enrolled at sites in Wuhan, China, and the sample size of each group was 20. In Groups A, B and C, the clinical remission rates were 95.0%%(19/20), 100.0%%(20/20) and 100.0%%(20/20), respectively. Compared with Groups A and B, the clinical remission time of Group C was significantly shorter (5.9 days vs. 10.8 days, p < 0.05; 5.9 days vs. 9.7 days, p < 0.05). There was no significant difference among Groups A, B, and C in terms of the time taken to be released from quarantine. The clinical biochemical indicators and safety indexes showed no significant differences among the three groups. CONCLUSIONS: Our findings suggest that Lopinavir-Ritonavir has some efficacy in the treatment of COVID-19, and the Huashi Baidu Formula might enhance this effect to an extent. In addition, superiority was displayed in the treatment of COVID-19 through a combination of the Huashi Baidu Formula and traditional Chinese medicine injection. In future, well-designed prospective double-blinded randomised control trials are required to confirm our findings.

A sensitive BODIPY-based fluorescent probe for detecting endogenous hydroxyl radicals in living cells.

The hydroxyl radical (˙OH) has been suggested to play very vital roles in many physiological and pathological processes. However, selective detection of ˙OH is highly challenging owing to its extremely high reactivity and short lifetime. Herein, we designed and synthesized a sensitive "turn on" fluorescent probe for detecting endogenous ˙OH based on a BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene) platform. The probe had shown good properties including high sensitively, ideal selectively and low cytotoxicity, and was successfully employed to image endogenous hydroxyl radical in living cells.

CdII -Organic Frameworks Fabricated with a N-Rich Ligand and Flexible Dicarboxylates: Structural Diversity and Multi-Responsive Luminescent Sensing for Toxic Anions and Ethylenediamine.

Three metal-organic frameworks {[Cd(L)(glu)]⋅3 H2 O}∞ (1), {[Cd2 (L)2 (adi)2 ]⋅5 H2 O}∞ (2) and {[Cd(L)(sub)]⋅3 H2 O⋅DMA }∞ (3) (L=pyridine-3,5-bis(5-azabenzimidazole), H2 glu=glutaric acid, H2 adi=adipic acid and H2 sub=suberic acid) were obtained under solvothermal conditions. Complex 1 shows a 2D (4,4) network constructing of Cd2 -glu and Cd-L chains. Complex 2 presents a 2-fold interpenetrating 3D framework with pcu topology. Complex 3 is a 3D framework with cds topology. Three complexes with versatile structures were obtained by changing aliphatic dicarboxylate ligands with different lengths based on a N-rich ligand. Moreover, the fluorescence measurements indicate that complex 1 is a good multifunctional chemosensor for the detection of Cr2 O7 2- and MnO4 - anions by fluorescence quenching effect, and ethylenediamine by fluorescence enhancement effect, with detection limits of 1.196 ppm, 0.551 ppm and 64.572 ppm, respectively. Both complexes 2 and 3 can selectively sense Cr2 O7 2- anion with detection limits of 1.126 ppm for 2 and 0.831 ppm for 3 by a fluorescence quenching effect.

A red fluorescent BODIPY probe for iridium (III) ion and its application in living cells.

A new red fluorescent probe 1 based on BODIPY skeleton has been successfully synthesized through introduction of 2-(thiophen-2-yl) quinoline moiety at meso- and 3-position, which exhibits excellent optical performance, including high fluorescence quantum yield, large pseudo Stokes' shift as well as high selectivity and sensitivity towards iridium (III) ion in aqueous solution and in living cells.

All-Organic Conductive Biomaterial as an Electroactive Cell Interface.

Various attractive materials are being used in bioelectronics recently. In this paper, hydroxymethyl-3,4-ethylenedioxythiophene (EDOT-OH) has been in situ integrated and polymerized on the surface of the regenerated silk fibroin (RSF) film to construct a biocompatible electrode. In order to improve the efficiency of in situ polymerization, sodium dodecyl sulfate (SDS) was adopted as surfactant to construct a well-organized and stable poly(hydroxymethyl-3,4-ethylenedioxythiophene) (PEDOT-OH) coating, whereas ammonium persulfate was used as oxidant. The effects of dosages of surfactant and oxidant, initial pH value, and monomer concentration on the polymerization were studied. Under the optimal conditions, the RSF/PEDOT-OH film exhibited a square resistance of 3.28 × 105 Ω corresponding to a conductance of 6.1 × 10-3 S/cm. Scanning electron microscope images indicated that PEDOT-OH was deposited uniformly on the surface of the RSF film with SDS. Furthermore, Fourier transform infrared spectroscopy confirmed that interactions existed between the peptide linkages of silk fibroin (SF) macromolecules and PEDOT-OH. The RSF/PEDOT-OH film displayed favorable electrochemical stability, biocompatibility, and fastness. This study provides a feasible method to endow conductivity to RSF materials in various forms. In addition, the conductive layer and biocompatible silk substrate make the RSF/PEDOT-OH biomaterial highly suitable for potential applications in bioelectric devices, sensors, and tissue engineering.

K2S2O8-Promoted Aryl Thioamides Synthesis from Aryl Aldehydes Using Thiourea as the Sulfur Source.

Thiourea as a sulfur atom transfer reagent was applied for the synthesis of aryl thioamides through a three-component coupling reaction with aryl aldehydes and N,N-dimethylformamide (DMF) or N,N-dimethylacetamide (DMAC). The reaction could tolerate various functional groups and gave moderate to good yields of desired products under the transition-metal-free condition.

[Clinical application evaluation and revision suggestions of clinical practice guideline on traditional Chinese medicine therapy alone or combined with antibiotics for acute pharyngitis].

The purpose of this study is to analyze the obstructive factors of clinical practice guideline on traditional Chinese medicine therapy alone or combined with antibiotics for acute pharyngitis, and summarize the revision suggestions for follow-up revision and promotion. Clinical physicians from 181 hospitals in 27 regions of China were selected to complete the online questionnaire survey for statistical analysis of obstructive factors. We collected 501 copies of the applicability evaluation questionnaire and 503 copies of the application evaluation questionnaire. The obstructive factors mainly focused on limitation of the Guideline, inconvenience of access, particularity of primary medical structure and uneven distribution of surveyed subjects. As for amendments, it was suggested to improve the syndrome differentiation, indications, prescriptions, and add characteristic TCM therapies in Chinese medicine; it was suggested to clarify the time to use antibiotics in Western medicine. According to the results of this study, the relevant contents of the Guideline should be further improved so as to be better applied in clinical practice.

The Association between Short-term Exposure to Fine Particulate Matter and Outpatient Visit in Beijing, China.

BACKGROUND: We tried to investigate the effect of PM2.5 on daily counts of outpatient visits in the Guang'anmen Hospital to determine if short-term PM2.5 exposure with extremely high concentration affects cardiopulmonary function of Beijing residents. METHODS: Outpatient visits and PM2.5 data from 01/11/2011 to 03/31/2013 were extracted from the Guang'anmen Hospital and the American Embassy in Beijing, respectively. Followed by using a semi-parametric generalized additive model (GAM) with time dependent covariates, we analyzed the association between PM2.5 concentrations and daily count of outpatient visits on Day 0, 1, 2, 3, 4 and 5 of PM2.5 exposure. RESULTS: Overall, 284354 subjects were collected. There were significant associations of short-term PM2.5 exposures with outpatient visits for cardiopulmonary diseases (P<0.05). Specifically, a 10 µg/m3 increase in PM2.5 was positively associated with a 0.74% of increase in angina visit on the first day and 0.50% increased visit on the second day (P<0.05). With an increase in PM2.5, the cough and respiratory visits significantly decreased by 0.17% and 0.30% on the first day, respectively (P<0.05). However, there were significant positive associations of PM2.5 with increased cough and respiratory visits (increased by 0.17% and 0.10%, respectively) on the fifth day (P<0.05). CONCLUSION: Our association studies showed an instant effect of PM2.5 level on cardiovascular outpatient visit in the Guang'anmen Hospital in Beijing while a lag effect on respiratory outpatient visits.

K2CO3-promoted formation of aryl esters from primary aryl amides by the acyl-acyl exchange process.

A new acyl-acyl exchange reaction has been developed for the formation of aryl esters from primary aryl amides. The reaction could occur under mild reaction conditions with catalytic quantities of K2CO3, and could afford moderate to good yields of the desired products.

Synthesis of imides by palladium-catalyzed C-H functionalization of aldehydes with secondary amides.

An efficient palladium-catalyzed C-H functionalization of aldehydes with various N-substituted N-heteroarene-2-carboxamides has been developed for the synthesis of secondary imides. The reaction tolerates various functionalities, such as methoxy, fluoro, chloro, and bromo groups. A tentative radical mechanism for a Pd(II)/Pd(IV) catalytic cycle is proposed.

[Treating influenza patients of wind-heat affecting Fei syndrome by jinhua qinggan granule: a double-blinded randomized control trial].

OBJECTIVE: To assess the effect and safety of Jinhua Qinggan Granule (JHG) in treating influenza patients of wind-heat affecting Fei syndrome (WHAFS). METHODS: Totally 136 influenza patients of WHAFS were randomized by stratification into 3 groups, the high dose JHG group (44 cases, 10 g each time), the low dose JHG group (45 cases, 5 g JHG + 5 g placebo each time), and the placebo control group (47 cases, 10 g placebo each time). All medication was administered three times daily for 5 days. The fever disappearance time, the fever disappearance rate, efficacy of TCM syndrome, the disappearance rate of main symptoms and physical signs of flu, the negative rate of virus nucleic acid in the pharyngeal secretion, and safety indicators were assessed. RESULTS: The median fever disappearance time was 32.8 h (95% CI: 22.5-41.0 h) in the high dose JHG group, 26.0 h (95% CI: 14.5-36.5 h) in the low dose JHG group, 39.5 h (95% CI: 29.0-46.0 h) in the placebo control group. There was statistical difference in the median fever disappearance time between the low dose JHG group and the placebo control group (P = 0.011). Three days after treatment, the markedly effective rate of TCM symptoms in the low dose JHG group was 66.7%, higher than that of the placebo control group (38.3%), and its effective rate was superior to that of the high dose JHG group (P = 0.043). Five days after treatment, the recovery rate of the low dose JHG group (42.2%) was higher than that of the high dose JHG group (25.0%, P = 0.026) and that of the placebo control group (14.9%, P = 0.002). The markedly effective rate of the low dose JHG group (86.7%) was higher than that of the placebo control group (55.3%, P = 0.001). Similar effects were obtained in the low dose JHG group and the high dose JHG group, but slightly poor in partial indicators of the high dose JHG group. There was no statistical difference in adverse reaction among these three groups (P > 0.05). CONCLUSIONS: JHG was effective and safe in treating influenza patients of WHAFS. Routinely low dose was the optimal dosage of JHG.

Coupling reactions of heteroarenes with phosphites under silver catalysis.

A silver-catalyzed dehydrogenative cross-coupling reaction of substituted furans, thiophene, thioazole, and pyrrole 1a-e with dialkyl phosphites 2 was first developed to afford corresponding phosphonated products 3a-h with up to 89% yield and good regioselectivities. Moreover, an unprecedented coupling of various substituted pyridines 1f-k with dialkyl phosphites 2 using AgNO(3) as a catalyst and K(2)S(2)O(8) as an oxidant, followed by reduction with Na(2)S(2)O(3), was also realized to furnish desired pyridine phosphonates 3i-q in satisfactory yields with good regioselectivities.

[Effects of Chinese medicine internal-external combined therapy in treating chronic obstructive pulmonary disease in the stable period].

OBJECTIVE: To observe the clinical effects of Chinese Medicine internal-external combined therapy (consisting of orally taking Chinese drugs and external acupoint sticking) for treatment of chronic obstructive pulmonary disease (COPD) in the stable period. METHODS: One hundred and seventy-eight COPD patients were randomly assigned to two groups, the control group was treated with Atrovent Metered Dose Inhalation, oral taking Mucosolvan and Bailing Capsule, and acupoint sticking with dummy plaster; the treatment group was treated with ipratropium bromide aerosol, ambroxol hydrochloride, Chinese recipe prescribed according to syndrome differentiation, and combined with acupoint sticking with Xiaochuan Plaster applied in winters and summers (3 times in a season). All were treated for three months. Changes of the Chinese medicine syndrome scores, quality of life (QOL), and the pulmonary function in patients before and after treatment were observed. RESULTS: Scores of Chinese medicine syndrome and QOL in the treatment group were significantly improved after treatment with the effect better than those in the control group (P < 0.05), but the change of the pulmonary function was insignificant (P > 0.05). CONCLUSION: Chinese medicine internal-external combined therapy shows definite effect in treating COPD patients in the stable stage, it could distinctively alleviate clinical symptoms and improve the QOL of patients.