Site-Specific Covalent Immobilization of SMA-Stabilized ACE2 for SARS-CoV-2 Recognition and Drug Screening.

Membrane protein (MP)-based biomaterials have a wide range of applications in drug screening, antigen detection, and ligand-receptor interaction analysis. Traditional MP immobilization methods have the disadvantage of disordered protein immobilization orientation, leading to the shielded binding domain and unreliable binding pattern. Herein, we describe a site-specific covalent immobilization of MPs, which utilizes the styrene maleic acid (SMA) detergent-free extraction method of MPs as well as the covalent reaction between His-tag and divinyl sulfone (DVS). As an example, we covalently immobilized angiotensin-converting enzyme 2 (ACE2) on a cell membrane chromatography system (ACE2-His-SMALPs/CMC) in a site-specific manner and verified the specificity and stability of this system. This technique significantly improves the service life compared to the physisorption CMC column. The improved protein immobilization strategies of the ACE2-His-SMALPs/CMC system enable it to effectively recognize SARS-CoV-2 pseudoviral particles as well as detect viral particles in ambient air once combined with an aerosol collector; as a powerful ligand biosensor, the ACE2-His-SMALPs/CMC system was used to screen for compounds with anti-SARS-CoV-2 pseudovirus activity. In conclusion, the optimized MP immobilization strategy has been successfully applied to CMC technology, showing enhanced stability and sensitivity, which can provide an efficient and convenient membrane protein immobilization method for biomaterials.

Longitudinal Changes of Sensorimotor Resting-State Functional Connectivity Differentiate between Patients with Thalamic Infarction and Pontine Infarction.

Purpose. We investigated the disparate influence of lesion location on functional damage and reorganization of the sensorimotor brain network in patients with thalamic infarction and pontine infarction. Methods. Fourteen patients with unilateral infarction of the thalamus and 14 patients with unilateral infarction of the pons underwent longitudinal fMRI measurements and motor functional assessment five times during a 6-month period (<7 days, at 2 weeks, 1 month, 3 months, and 6 months after stroke onset). Twenty-five age- and sex-matched controls underwent MRI examination across five consecutive time points in 6 months. Functional images from patients with left hemisphere lesions were first flipped from the left to the right side. The voxel-wise connectivity analyses between the reference time course of each ROI (the contralateral dorsal lateral putamen (dl-putamen), pons, ventral anterior (VA), and ventral lateral (VL) nuclei of the thalamus) and the time course of each voxel in the sensorimotor area were performed for all five measurements. One-way ANOVA was used to identify between-group differences in functional connectivity (FC) at baseline stage (<7 days after stroke onset), with infarction volume included as a nuisance variable. The family-wise error (FWE) method was used to account for multiple comparison issues using SPM software. Post hoc repeated-measure ANOVA was applied to examine longitudinal FC reorganization. Results. At baseline stage, significant differences were detected between the contralateral VA and ipsilateral postcentral gyrus (cl_VA-ip_postcentral), contralateral VL and ipsilateral precentral gyrus (cl_VL-ip_precentral). Repeated measures ANOVA revealed that the FC change of cl_VA-ip_postcentral differ significantly among the three groups over time. The significant changes of FC between cl_VA and ip_postcentral at different time points in the thalamic infarction group showed that compared with 7 days after stroke onset, there was significantly increased FC of cl_VA-ip_postcentral at 1 month, 3 months, and 6 months after stroke onset. Conclusions. The different patterns of sensorimotor functional damage and reorganization in patients with pontine infarction and thalamic infarction may provide insights into the neural mechanisms underlying functional recovery after stroke.

Effect of Near-Infrared Pulsed Light on the Human Brain Using Electroencephalography.

In our previous study, the low-level laser (LLL) stimulation at the palm with a stimulation frequency of 10 Hz was able to induce significant brain activation in normal subjects. The electroencephalography (EEG) changes caused by the stimulation of light-emitting diode (LED) in normal subjects have not been investigated. This study aimed at identifying the effects of LED stimulation on the human brain using EEG analysis. Moreover, the dosage has been raised 4 times than that in the previous LLL study. The LED array stimulator (6 pcs LEDs, central wavelength 850 nm, output power 30 mW, and operating frequency 10 Hz) was used as the stimulation source. The LED stimulation was found to induce significant variation in alpha activity in the occipital, parietal, and temporal regions of the brain. Compared to the previous low-level laser study, LED has similar effects on EEG in alpha (8-12 Hz) activity. Theta (4-7 Hz) power significantly increased in the posterior head region of the brain. The effect lasted for at least 15 minutes after stimulation ceased. Conversely, beta (13-35 Hz) intensity in the right parietal area increased significantly, and a biphasic dose response has been observed in this study.

Rescue fecal microbiota transplantation for antibiotic-associated diarrhea in critically ill patients.

BACKGROUND: Antibiotic-associated diarrhea (AAD) is a risk factor for exacerbating the outcome of critically ill patients. Dysbiosis induced by the exposure to antibiotics reveals the potential therapeutic role of fecal microbiota transplantation (FMT) in these patients. Herein, we aimed to evaluate the safety and potential benefit of rescue FMT for AAD in critically ill patients. METHODS: A series of critically ill patients with AAD received rescue FMT from Chinese fmtBank, from September 2015 to February 2019. Adverse events (AEs) and rescue FMT success which focused on the improvement of abdominal symptoms and post-ICU survival rate during a minimum of 12 weeks follow-up were assessed. RESULTS: Twenty critically ill patients with AAD underwent rescue FMT, and 18 of them were included for analysis. The mean of Acute Physiology and Chronic Health Evaluation (APACHE) II scores at intensive care unit (ICU) admission was 21.7 ± 8.3 (range 11-37). Thirteen patients received FMT through nasojejunal tube, four through gastroscopy, and one through enema. Patients were treated with four (4.2 ± 2.1, range 2-9) types of antibiotics before and during the onset of AAD. 38.9% (7/18) of patients had FMT-related AEs during follow-up, including increased diarrhea frequency, abdominal pain, increased serum amylase, and fever. Eight deaths unrelated to FMT occurred during follow-up. One hundred percent (2/2) of abdominal pain, 86.7% (13/15) of diarrhea, 69.2% (9/13) of abdominal distention, and 50% (1/2) of hematochezia were improved after FMT. 44.4% (8/18) of patients recovered from abdominal symptoms without recurrence and survived for a minimum of 12 weeks after being discharged from ICU. CONCLUSION: In this case series studying the use of FMT in critically ill patients with AAD, good clinical outcomes without infectious complications were observed. These findings could potentially encourage researchers to set up new clinical trials that will provide more insight into the potential benefit and safety of the procedure in the ICU. TRIAL REGISTRATION: ClinicalTrials.gov, Number NCT03895593 . Registered 29 March 2019 (retrospectively registered).

Neuronal Specificity of Acupuncture in Alzheimer's Disease and Mild Cognitive Impairment Patients: A Functional MRI Study.

Although acupuncture is considered to be effective and safe for Alzheimer's disease (AD) and mild cognitive impairment (MCI), the mechanism underlying its therapeutic effect is still unknown. Most studies clarifying the neuronal pathway produced by acupuncture were still applied to healthy subjects with limited single acupuncture point stimulation, which was inconsistency with clinical practice. Thus, in our present study, we investigate the differences between brain activity changes in AD and MCI patients caused by multi-acupuncture point Siguan (four gates), in order to provide visualized evidence for neuronal specificity of clinical acupuncture. Forty-nine subjects were recruited, including 21 AD patients, 14 MCI patients, and 14 healthy controls (HC). AD and MCI patients were randomly divided into two groups, respectively: real acupuncture point group (14 AD and 8 MCI) and sham acupuncture point group (7 AD and 6 MCI). We adopted a 16-minute, single-block, experimental design for acquiring functional MRI images. We found, in AD and MCI patients, Siguan (four gates) elicited extensive activations and deactivations in cognitive-related areas, visual-related areas, the sensorimotor-related area, basal ganglia, and cerebellum. Compared with HC, AD and MCI patients showed similar activations in cognitive-related brain areas (inferior frontal gyrus, supramarginal gyrus, and rolandic operculum) as well as deactivations in cognitive-related areas, visual-related areas, basal ganglia, and cerebellum, which were not found in HC. Compared with sham acupuncture points, real acupuncture points produced more specific brain changes with both activated and deactivated brain activities in AD and MCI. The preliminary results in our study verified the objective evidence for neuronal specificity of acupuncture in AD and MCI patients.

Baicalein improves behavioral dysfunction induced by Alzheimer's disease in rats.

BACKGROUND: Alzheimer's disease (AD) is considered to be a neurodegenerative disorder that is characterized by increased oxidative stress. Medicinal plants, with their antioxidant properties, have been used to cure several human diseases. The aim of the current study was to explore the protective and therapeutic effect of baicalein on AD-induced rats. MATERIALS AND METHODS: Swiss Wistar rats were used in the study. The rats were divided into five groups. Group I: normal control group treated with water; Group II: disease control treated with AlCl3 to induce the mimicking AD for 4 successive weeks (SW); Group III: normal control group treated with baicalein (5 mg/kg) for 2 SW followed by combination of baicalein and AlCl3 for 4 SW; Group IV: normal control group treated with baicalein (10 mg/kg) for 2 SW followed by combination of baicalein and AlCl3 for 4 SW; Group V: normal control group treated with rivastigmine (0.3 mg/kg) for 2 SW followed by combination of rivastigmine and AlCl3 for 4 SW. Moreover, the therapeutic groups are as follows: Group VI: AD disease control treated with AlCl3 for 4 SW and serving as the therapeutic positive group; Group VII: AD disease control + baicalein (5 mg/kg) for 12 SW; Group VIII: AD disease control + baicalein (10 mg/kg) for 12 SW; Group IX: AD disease control + rivastigmine (0.3 mg/kg) for 12 SW. Behavioral test, T-maze, and rotarod test were also performed before and after the treatment. At the end of the experimental study, all the rats were sacrificed and their brains were removed and divided into two portions. The first portion was homogenated for estimating the level of acetylcholinesterase (AchE) and acetylcholine (Ach). Another portion was used for histopathological evaluation. RESULTS: The current investigation showed that baicalein significantly reduced the duration of revolving on the rotarod, cage activity, and T-maze activity in a dose-dependent manner compared with the AD control group rats. It also altered the AchE and Ach levels in the brain homogenates. The histopathology study also provides strength to the protective effect of baicalein. CONCLUSION: The current study showed that baicalein significantly (P<0.05) improved the biochemical and histopathological condition of AD in rats.

Inflammatory morphea mimicking an acquired port-wine stain initially treated with pulsed-dye laser.

The early inflammatory lesions of morphea may present with erythema or violaceous patches and plaques before evolving into areas of sclerosis. They have been misdiagnosed as acquired port-wine stains (PWSs). We report a previously well 7-year-old Chinese girl presenting with early facial morphea mimicking an acquired PWS with unusual histologic features of perineural inflammation. The presence of cutaneous perineural inflammation may be seen in a small percentage of cases of morphea and appears to be a feature of early inflammatory morphea. We report this case to highlight the importance in recognizing this entity and summarize the reported cases of inflammatory morphea mimicking acquired PWSs.

An FMRI study of neuronal specificity in acupuncture: the multiacupoint siguan and its sham point.

Clarifying the intrinsic mechanisms of acupuncture's clinical effects has recently been gaining popularity. Here, we choose the Siguan acupoint (a combination of bilateral LI4 and Liv3) and its sham point to evaluate multiacupoint specificity. Thirty-one healthy volunteers were randomly divided into real acupoint (21 subjects) and sham acupoint (10 subjects) groups. Our study used a single block experimental design to avoid the influence of posteffects. Functional magnetic resonance imaging data were acquired during acupuncture stimulation. Results showed extensive increase in neuronal activities with Siguan acupuncture and significant differences between stimulation at real and sham points. Brain regions that were activated more by real acupuncture stimulation than by sham point acupuncture included somatosensory cortex (the superior parietal lobule and postcentral gyrus), limbic-paralimbic system (the calcarine gyrus, precuneus, cingulate cortex, and parahippocampal gyrus), visual-related cortex (the fusiform and occipital gyri), basal ganglia, and the cerebellum. In this way, our study suggests Siguan may elicit specific activities in human brain.

[HPLC fingerprint of glycyrrhizea radix et rhizoma praeparata cum melle].

The chromatographic fingerprint was established by eluting with the mobile phase consisted of acetonitrile and 0.2% formic acid water on an Agilent TC-C18 (2) column (4.6 mm x 250 mm, 5 microm). Six chromatographic peaks were identified by HPLC-MS/MS method. Ten batches of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle were determined, and the similarity was arranged from 0.72 to 0.99. Good precision, stability and repeatability were obtained, and this study provides a reference for the quality control of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle.

Danggui-Shaoyao-San Improves Learning and Memory in Female SAMP8 via Modulation of Estradiol.

Previous studies showed that Danggui-Shaoyao-San (DSS), a traditional Chinese medicinal prescription, could alleviate cognitive dysfunction of Alzheimer's disease (AD) patients. However, the mechanisms remain unclear; we have now examined the effect of DSS on SAMP8 and elucidated the possible mechanism. Animals were treated with DSS for 2 months, and step-down test and Morris water maze (MWM) test were used to evaluated cognitive abilities. The estradiol (E2), NO, and glycine in blood plasma or in hippocampus were detected to explore the possible mechanisms. The latency of SAMP8 in step-down test was shorter than that of age-matched SAMR1, and DSS increased the latency especially in female animals. In MWM test, we got similar results; SAMP8 spent more time to find the platform, and DSS decreased the time before finding the platform, with little effect on swim velocity, during the training sessions. During test session, DSS increased the time spent in target quadrant especially in female SAMP8. In female SAMP8, plasma E2, NO, and glycine were elevated in plasma or hippocampus tissue. In conclusion, DSS could ameliorate deterioration of cognition in SAMP8, especially in female animals. Increasing E2, NO, and glycine might contribute to the cognitive improvement effect of DSS in female SAMP8.

HPLC fingerprint of glycyrrhizea radix et rhizoma praeparata cum melle / 中国中药杂志

The chromatographic fingerprint was established by eluting with the mobile phase consisted of acetonitrile and 0.2% formic acid water on an Agilent TC-C18 (2) column (4.6 mm x 250 mm, 5 microm). Six chromatographic peaks were identified by HPLC-MS/MS method. Ten batches of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle were determined, and the similarity was arranged from 0.72 to 0.99. Good precision, stability and repeatability were obtained, and this study provides a reference for the quality control of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle.

Anti-apoptotic effect of Astragalus Polysaccharide on myeloid cells / 中国实验血液学杂志

This study was aimed to assess the effect of Astragalus Polysaccharide (ASPS) on in-vitro hematopoiesis. CFU-GM assays were used to determine the effect of ASPS and thrombopoietin (TPO) on granulocytic-monocyte progenitor cells. The CFU assays were also used to investigate the effect of ASPS on the proliferation of HL-60 cells.HL-60 cells were cultured with serum-free RPMI 1640 medium and treated with or without of different concentrations of ASPS. After 72 h incubation, the number of cells were counted.In addition, the caspase-3 and JC-1 expression was determined by flow cytometry with Annexin V/PI double staining. The results showed that ASPS (100, 200 µg/ml) and TPO (100 ng/ml) significantly promoted CFU-GM formation in vitro. Various concentrations of ASPS and TPO also promoted the colony formation of HL-60 cells, the largest effect of ASPS was observed at a concentration of 100 µg/ml. There were no synergistic effects between TPO and ASPS on cellular proliferation. The results also showed that ASPS significantly protected HL-60 cells from apoptosis in condition of serum-free medium culture, suppressed caspase 3 activation, and reduced the cell apoptosis. It is concluded that ASPS can significantly promote the formation of bone marrow CFU-GM and the proliferation of HL-60 cells, the optimal concentration of ASPS is at 100 µg/ml. In the absence of serum inducing apoptosis, ASPS also significantly reduced the apoptosis of HL-60 cells via suppressing the activation of caspase-3.

[Chemical constituents of glucoside fraction from Liuwei Dihuang Gantang].

OBJECTIVE: To study chemical constituents of glucoside fraction from Liuwei Dihuang Gantang and clarify its substance foundation of active constituents. METHOD: Glucoside fraction was prepared by macroporous resin chromatography. Constituents were separated by silica gel and reverse phase silica gel column chromatography, and their structures were identified by MS and NMR. RESULT: Eleven compounds were separated and identified as 7-dehydrologanin (1), 7alpha-O-methylmorroniside (2), 7beta-O-methylmorroniside (3), 7alpha-O-ethylmorroniside (4), 7beta-O-ethylmorroniside (5), morroniside (6), sweroside (7), loganin (8), paeoniflorin (9), paeonolide (10) and loganic acid (11). CONCLUSION: All of those compounds were separated from Liuwei Dihuang Gantang for the first time.

Effect of low-level laser stimulation on EEG.

Conventional laser stimulation at the acupoint can induce significant brain activation, and the activation is theoretically conveyed by the sensory afferents. Whether the insensible low-level Laser stimulation outside the acupoint could also evoke electroencephalographic (EEG) changes is not known. We designed a low-level laser array stimulator (6 pcs laser diode, wavelength 830 nm, output power 7 mW, and operation frequency 10 Hz) to deliver insensible laser stimulations to the palm. EEG activities before, during, and after the laser stimulation were collected. The amplitude powers of each EEG frequency band were analyzed. We found that the low-level laser stimulation was able to increase the power of alpha rhythms and theta waves, mainly in the posterior head regions. These effects lasted at least 15 minutes after cessation of the laser stimulation. The amplitude power of beta activities in the anterior head regions decreased after laser stimulation. We thought these EEG changes comparable to those in meditation.

[HPLC fingerprint of Liuwei Dihuang condensed pills].

OBJECTIVE: To establish HPLC fingerprints of Liuwei Dihuang condensed pills. METHOD: Dikma Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) was adopted, with acetonitrile (containing 0.05% phosphoric) -water (containing 0.05% phosphoric) as the mobile phase. The column temperature was set at 40 degrees C, and the flow rate was 1.0 mL x min(-1). The detection wavelength was 276 nm (0-10 min), 236 nm (10-40 min) and 276 nm (40-60 min). The sample size was 20 microL. Chromatographic peaks were identified by Q-TOF-MS-IDA-MS/MS method. RESULT: Good precision, stability and repeatability were proved. Q-TOF-MS-IDA-MS/ MS method was adopted for qualitative determination of eighteen chromatographic peaks. Ten batches of Liuwei Dihuang condensed pills were determined with the method, and their similarities were above 0. 96. CONCLUSION: The study lays a foundation for the overall quality evaluation of Liuwei Dihuang condensed pills.

JD-30, an active fraction extracted from Danggui-Shaoyao-San, decreases ß-amyloid content and deposition, improves LTP reduction and prevents spatial cognition impairment in SAMP8 mice.

JD-30 is an active fraction extracted from Danggui-Shaoyao-San (DSS), a traditional Chinese medicinal prescription. We previously showed that JD-30 could alleviate cognitive dysfunction of the mice induced by intracerebroventricular injection of ß-amyloid (Aß). However, data remain scarce on the effect of JD-30 on an Alzheimer's disease (AD) model and the underlying mechanisms are unknown. Further detailed studies on the effects of JD-30 on spatial cognition of senescence-accelerated mouse prone 8 (SAMP8), a suitable rodent model for cognitive impairment of aged subjects were investigated to elucidate the possible mechanisms. Long-term treatment with JD-30 significantly decreased the prolonged latency of SAMP8 in the Morris water-maze test. It also ameliorated the reduction of long-term potentiation (LTP) and reduced the damage of neurons in the hippocampus of SAMP8. Finally, JD-30 decreased the content and deposition of Aß in the brain of SAMP8. The results show that JD-30 improves deterioration of spatial learning and memory in the SAMP8 mouse model, and by decreasing the content and deposition of Aß, neuronal activity and synaptic plasticity improve, suggesting one of the mechanisms involved.

Chemical constituents of glucoside fraction from Liuwei Dihuang Gantang / 中国中药杂志

<p><b>OBJECTIVE</b>To study chemical constituents of glucoside fraction from Liuwei Dihuang Gantang and clarify its substance foundation of active constituents.</p><p><b>METHOD</b>Glucoside fraction was prepared by macroporous resin chromatography. Constituents were separated by silica gel and reverse phase silica gel column chromatography, and their structures were identified by MS and NMR.</p><p><b>RESULT</b>Eleven compounds were separated and identified as 7-dehydrologanin (1), 7alpha-O-methylmorroniside (2), 7beta-O-methylmorroniside (3), 7alpha-O-ethylmorroniside (4), 7beta-O-ethylmorroniside (5), morroniside (6), sweroside (7), loganin (8), paeoniflorin (9), paeonolide (10) and loganic acid (11).</p><p><b>CONCLUSION</b>All of those compounds were separated from Liuwei Dihuang Gantang for the first time.</p>

HPLC fingerprint of Liuwei Dihuang condensed pills / 中国中药杂志

<p><b>OBJECTIVE</b>To establish HPLC fingerprints of Liuwei Dihuang condensed pills.</p><p><b>METHOD</b>Dikma Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) was adopted, with acetonitrile (containing 0.05% phosphoric) -water (containing 0.05% phosphoric) as the mobile phase. The column temperature was set at 40 degrees C, and the flow rate was 1.0 mL x min(-1). The detection wavelength was 276 nm (0-10 min), 236 nm (10-40 min) and 276 nm (40-60 min). The sample size was 20 microL. Chromatographic peaks were identified by Q-TOF-MS-IDA-MS/MS method.</p><p><b>RESULT</b>Good precision, stability and repeatability were proved. Q-TOF-MS-IDA-MS/ MS method was adopted for qualitative determination of eighteen chromatographic peaks. Ten batches of Liuwei Dihuang condensed pills were determined with the method, and their similarities were above 0. 96.</p><p><b>CONCLUSION</b>The study lays a foundation for the overall quality evaluation of Liuwei Dihuang condensed pills.</p>

Danggui-Shaoyao-San and its active fraction JD-30 improve Abeta-induced spatial recognition deficits in mice.

AIMS OF THE STUDY: Previous studies showed that Danggui-Shaoyao-San (DSS), a traditional Chinese medicinal prescription, could alleviate cognitive dysfunction of Alzheimer's disease (AD) patients. However, the mechanism and substance basis remain unknown. JD-30 is a fraction extracted from DSS, whose activity we previously was evaluated. beta-Amyloid (Abeta) is believed to be a critical etiological factor of AD. We have now examined the effect of DSS and JD-30 on AD model mice induced by Abeta, and elucidated the possible mechanism. MATERIALS AND METHODS: Mice were intracerebroventricular injected with the aggregated Abeta(25-35) to mimic AD. Groups of mice were treated with DSS or JD-30 by intragastric infusion over 2 weeks, and their spatial learning and memory capacities were measured by the Morris water maze procedure. The mechanisms were investigated by extracellular microelectrode recordings, and also electron microscopy. RESULTS: Our results show that Abeta(25-35) induced impairment of spatial learning and memory in mice, as well as inhibition of long-term potentiation (LTP) in the hippocampus. The impairments were ameliorated by DSS or JD-30 administration. Additionally, JD-30 not only prevented the aggregation of Abeta(25-35), but disrupted aggregated Abeta(25-35) fibrils. CONCLUSION: These results suggest that JD-30 is one of the chief active fractions extracted from DSS by its ability to ameliorate deterioration of cognition, and by blocking and disrupting the aggregation of Abeta so that synaptic plasticity was improved, which may be one of the mechanisms involved.

An original non-traumatic maneuver for repositioning migrated peritoneal dialysis catheters.

OBJECTIVE: To describe an original non-traumatic maneuver for repositioning a migrated peritoneal dialysis (PD) catheter. METHODS: First, tissues wrapping the PD catheter are dissociated, then a correct route is selected according to the position of the catheter, and finally the catheter is repositioned manually in 8 steps: pressing, palpating, vibrating, wave vibrating, rotating, back-pushing and vibrating, swaying, and compressing. RESULTS: Of 30 cases of PD catheter migration, repositioning was successful on the first attempt in 9 cases, on the third attempt in 10 cases, on the seventh attempt in 7 cases, and failed in 4 cases. The overall success rate was 86.7%. CONCLUSION: Manual repositioning of a migrated PD catheter is safe, painless, economical, and feasible. Repositioning of the migrated dialysis catheter may be attempted before referral for more invasive interventions.