A rapid and sensitive UPLC-MS/MS method for the determination of zanubrutinib in beagle plasma and its application in pharmacokinetics.

A reliable and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the determination of zanubrutinib in the plasma of beagle dogs. The column used was an Acquity BEH C18 column (2.1 mm × 50 mm, 1.7 µm), maintained at 40°C with an injection volume of 2 µl. The gradient elution program was as follows: 0-1 min, 10-10% A; 1-1.1 min, 10-90% A; 1.1-2.1 min, 90-90% A; 2.1-2.2 min, 90-10% A; 2.2-3.0 min, 10-10% A. Mobile phase A was 0.1% formic acid, B was acetonitrile, and the total analysis time was 3 min. The mass spectrometry was performed in positive ion mode, and the scanning mode was multi-reaction monitoring mode with electrospray ionization as the ion source; m/z 472.2 → 455.01 for zanubrutinib and m/z 441.03 → 137.99 for ibrutinib (internal standard). The plasma samples were processed by protein precipitation. The standard curve showed good linearity (r2 = 0.999 8) in the range of 1.0-1,000 ng/ml (zanubrutinib) with a low limit of quantification of 1 ng/ml. Also, the intra-day and inter-day precision (RSD) was <5.88% and the accuracy (RE) ranged from -1.56 to 1.08%; the recoveries of zanubrutinib in beagle plasma ranged from 90.12 to 93.53% (RSD 1.67-6.42%) and the ME values of zanubrutinib were 98.70-101.06% (RSD 5.37-8.49%, n = 6). All values meet US Food and Drug Administration requirements. A rapid, highly selective and sensitive method for the determination of zanubrutinib concentration in plasma by UPLC-MS/MS was successfully developed. This method is suitable for pharmacokinetic studies in beagle dogs by following oral administration of zanubrutinib.

Enhanced mass transfer of pulsed vacuum pressure pickling and changes in quality of sour bamboo shoots.

Sour bamboo shoot is a traditional Chinese fermented vegetable food. The traditional pickling method of sour bamboo shoots has the disadvantages of being time-consuming, inhomogeneous, and difficult to control. Pulsed vacuum pressure pickling (PVPP) technology uses pulsed vacuum pressure to enhance the pickling efficiency significantly. To demonstrate the effects of salt content and PVPP technical parameters on the fermentation of bamboo shoots, the sample salinity, pH value, color, crunchiness and chewiness, nitrite content, and lactic acid bacteria content during the pickling process were investigated. The salt content inside the bamboo shoots gradually increased to the equilibrium point during the pickling process. The pickling efficiency of bamboo shoots under PVPP technology increased by 34.1% compared to the traditional control groups. Meanwhile, the uniform salt distribution under PVPP technology also obtained better performance in comparison with the traditional groups. The pH value declined slowly from 5.96 to 3.70 with the extension of pickling time and sour flavor accumulated progressively. No significant differences were found in the color values (L *, a *, and b *) and the crunchiness of the bamboo shoot under different salt solution concentrations, vacuum pressure, and pulsation frequency ratio conditions. Colony-forming unit of lactic acid bacteria (CFU of LAB) decreased, to begin with, and then increased until the 6th day, followed by a declining trend in volatility. The nitrate content of bamboo shoots samples under PVPP treatments did not exceed the safety standard (<20 mg/kg) during the whole fermentation process, which proves the safety of PVPP technology. In conclusion, PVPP technology can safely replace the traditional method with better quality performance. The optimal PVPP processing conditions (vacuum pressure 60 kPa, 10 min vacuum pressure time vs. 4 min atmospheric pressure time, salt solution concentration 6%) have been recommended for pickling bamboo shoots with high product quality.

Twelve-year profile of screening results of retinopathy of prematurity at a tertiary care institute in Northwest China.

AIM: To report the changes in detection rate and characteristics of retinopathy of prematurity (ROP) in infants, during a 12-year period in Northwest China. METHODS: The medical records of infants were retrospectively collected and reviewed using an established clinical database. The detection rate and severity of ROP were compared between two consecutive periods (P1: 2008-2013, P2: 2014-2019). Gender, gestational age (GA), birth weight (BW), multiple births, delivery pattern, and postmenstrual age of the first fundus screen were analyzed in all visiting infants. RESULTS: During the 12-year study period, 7832 infants were initially included; among them, 1266 (16.16%) were diagnosed with ROP, 441 of whom (5.63%) developed severe ROP. Throughout the study period, the total number of infants being screened showed a trend of slight fluctuation after a rapid increase; however, an annual increase was observed in the number of infants diagnosed with ROP and severe ROP. The proportion of each stage at the first screening of infants with ROP was stable. The detection rate of ROP increased from 2.33% in 2008 to 16.18% in 2010, decreased to 10.73% in 2014, and then increased to 27.47% in 2019. For severe ROP, the detection rate gradually increased from 0 in 2008 to 12.49% in 2019. Among the infants with ROP, 96 (7.58%) did not meet the screening criteria set by the Chinese Medical Association in 2014 (GA<32wk, or BW<2000 g); among them, 14 (1.11%) needed treatment because of severe ROP. CONCLUSION: From 2008 to 2019, the detection rates of ROP and severe ROP in infants screened in Northwest China were 16.16% and 5.63%, respectively. The characteristics of the ROP infants were similar to those in other middle-income regions. The "tertiary prevention network of ROP" is a potentially effective screening approach.

The Pharmacokinetic Effect of Itraconazole and Voriconazole on Ripretinib in Beagle Dogs by UPLC-MS/MS Technique.

BACKGROUND: A new UPLC-MS/MS technique for the determination of ripretinib in beagle dog plasma was developed, and the pharmacokinetic effects of voriconazole and itraconazole on ripretinib in beagle dogs were studied. METHODS: After extraction with ethyl acetate under alkaline conditions, ripretinib was detected using avapritinib as the internal standard (IS). The mobile phases were 0.1% formic acid-acetonitrile. The scanning method was multi-reaction monitoring using ESI+ source, and the ion pairs for ripretinib and IS were m/z 509.93→416.85 and 499.1→482.09, respectively. This animal experiment adopted a three period self-control experimental design. In the first period, ripretinib was orally administered to six beagle dogs at a dose of 5 mg/kg. In the second period, the same six beagle dogs were orally given itraconazole at a dose of 7 mg/kg, after 30 min, ripretinib was orally given. In the third period, voriconazole at a dose of 7 mg/kg was given orally, and then ripretinib was orally given. At different time points, the blood samples were collected. The concentration of ripretinib was detected, and the pharmacokinetic parameters of ripretinib were calculated. RESULTS: Ripretinib had a good linear relationship in the range of 1-1000 ng/mL. The precision, accuracy, recovery, matrix effect and stability met the requirements of the guiding principles. After erdafitinib combined with itraconazole, the Cmax and AUC0→t of ripretinib increased by 38.35% and 36.36%, respectively, and the t1/2 was prolonged to 7.53 h. After ripretinib combined with voriconazole, the Cmax and AUC0→t of ripretinib increased by 37.44% and 25.52%, respectively, and the t1/2 was prolonged to 7.33 h. CONCLUSION: A new and reliable UPLC-MS/MS technique was fully optimized and developed to detect the concentration of ripretinib in beagle dog plasma. Itraconazole and voriconazole could inhibit the metabolism of ripretinib in beagle dogs and increase the plasma exposure of ripretinib.

Traditional uses, phytochemistry, and pharmacology of Persicaria orientalis (L.) Spach - A review.

ETHNOPHARMACOLOGICAL RELEVANCE: Persicaria orientalis (L.) Spach (internationally accepted and only valid name; synonym: Polygonum orientale L.; family: Polygonaceae), which is named Hongcao in China, is a Chinese herbal medicine that has a wide range of pharmacological effects including treatment to rheumatoid arthritis, coronary heart disease, hernia, carbuncle sore, enhance immunity, antimicrobial, osteogenic and dilated bronchiectasis. AIM OF THIS REVIEW: This review aims to provide systematically organized information on traditional uses of Persicaria orientalis (L.) Spach (P. orientalis) and to critically analyze evidences in phytotherapeutic, botanical, and pharmacological literatures that support its therapeutic potential in treatment to human diseases. Isolation of additional compounds and detailed pharmacological investigations are key areas to investigate. MATERIALS AND METHODS: Relevant information on P. orientalis was collected through published scientific materials (including PubMed, ScienceDirect, Wiley, ACS, CNKI, Scifinder, Springer, Taylor & Francis, Web of Science, Google Scholar, and Baidu Scholar) and other literature sources (e.g., Chinese Pharmacopoeia, 2015 edition, Chinese herbal classic books and PhD and MSc thesis, etc.). RESULTS: Traditional uses were compiled in this review, including classic prescriptions and historical applications. Approximately 70 compounds, mainly including flavonoids, phenolics, lignans, limonoids and steroids, have been isolated and identified from P. orientalis. Among them, flavonoids were main components. Crude extracts and pure compounds isolated from P. orientalis exhibited various pharmacological activities, such as protection against ischemia and hypoxia-induced myocardial cells and hypoxia/reoxygenation cardiomyocyte, increase the blood flow in myocardium, expanding bronchus, anti-inflammatory and analgesic, and antithrombotic effects and so on. CONCLUSIONS: P. orientalis is a valuable source with therapeutic potential on a wide range of diseases especially cardiovascular-system disorders. Though most traditional uses of P. orientalis are supported by in vitro/vivo pharmacological studies, however, there is still a lack of researches on active pharmacodynamic ingredients as well as in-depth and in-vivo mechanistic studies. Therefore, isolation and identification of more active compounds (especially flavonoids), their structure-activity relationship and studies on pharmacodynamic mechanisms by more elaborative in-vivo studies on P. orientalis may be focused on in order to confirm efficacy of reported therapeutic effects of P. orientalis and help explore it's therapeutic potentials. Furthermore, research designs of pharmacological studies based on traditional uses of anti-rheumatoid arthritis through cell lines and animal models should also be considered as key research topics.

Anti-rheumatoid arthritis effects in adjuvant-induced arthritis in rats and molecular docking studies of Polygonum orientale L. extracts.

BACKGROUND AND PURPOSE: Polygonum orientale L. (family: Polygonaceae), named Hongcao in China, has effects of dispelling wind and dampness, promoting blood circulation, and relieving pain. Our group has already studied and confirmed that POEa and POEe (ethyl acetate and ethyl ether extract of P. orientale, respectively) had anti-inflammatory and analgesic effects in early research, which was mainly relevant to the existence of flavonoids. According to the clinical application of P. orientale in traditional Chinese medicine, it has long been used for rheumatic arthralgia and rheumatoid arthritis. Therefore, our group further explored whether flavonoids of P. orientale have anti-rheumatoid arthritis effect and how does they play this role. METHODS: Dried small pieces of the stems and leaves of P. orientale were decocted with water and partitioned successively to obtain POEa and POEe, respectively. The anti-rheumatoid arthritis effect of P. orientale was studied by using a Freund's complete adjuvant (FCA)-induced arthritis (AIA) in a rat model. The levels of PGE2, TNF-α, and IL-1ß in serum of AIA rats were detected by enzyme linked immunosorbent assay (ELISA) to explore its mechanisms. In addition, we computationally studied the relationships between the 15 chemical components of POEa and POEe, and the currently focused 9 target proteins of rheumatoid arthritis by molecular docking. RESULTS: Pharmacological experiments showed that POEa and POEe significantly ameliorate symptoms of rheumatoid arthritis via reducing paw swelling volume, arthritis score, and thymus and spleen indices, as well as increasing body weight in AIA rats. Simultaneously, the concentrations of PGE2, TNF-α, and IL-1ß were significantly decreased by POEa and POEe. Histopathology revealed noticeable reduction in bone and cartilage, synovial hyperplasia, inflammatory cell infiltration, cartilage surface erosion, and joint degeneration by POEa and POEe treatment. In addition, the molecular docking studies showed that docking scores of 14 chemical compositions (including 12 flavonoids and 2 phenolic acids) of POEa and POEe with anti-rheumatoid arthritis protein targets were better than the complexed ligands of the anti-rheumatoid arthritis protein targets. Among them, six flavonoids in POEa and POEe had more docking protein targets (n ≥ 3). Five anti-rheumatoid arthritis targets including high-temperature requirement A1 protease (HtrA1), janus kinase 1 (JAK1), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (i-NOS), and prostaglandin E2 (PGE2) had better docking score compared with the complexed ligands. Moreover, most of the chemical components in POEa and POEe showed strong interaction with HtrA1. CONCLUSIONS: The flavonoids of P. orientale have anti-rheumatoid arthritis effect. In addition, the molecular docking results indicate that quercetin, catechol, orientin, and other six flavonoids may be closely related to HtrA1, JAK1, COX-2, i-NOS, and PGE2 protein target receptors. It suggests that these chemical compositions form strong protein-ligand complexes with these protein targets, especially HtrA1 to exert anti-rheumatoid arthritis. Further experimental studies show that mechanisms of anti-rheumatoid arthritis effects may also be relevant to inhibit the levels of PGE2, TNF-α, and IL-1ß in serum. Therefore, our group can further explore the possible active ingredients and mechanisms of the anti-rheumatoid arthritis effects of flavonoids, and focus on the inhibition of the expression of inflammatory factors and the TGF-ß1/Smad signaling pathway associated with HtrA1 protein target receptors, which can provide a direction and powerful reference for the action mechanism and drug research of anti-rheumatoid arthritis of flavonoids in P. orientale.

Anti-inflammatory and Analgesic Effects of Polygonum orientale L. Extracts.

Background and Purpose:Polygonum orientale L. (family: Polygonaceae), named Hongcao in China, is a Traditional Chinese Medicinal and has long been used for rheumatic arthralgia and rheumatoid arthritis. However, no pharmacological and mechanism study to confirm these clinic effects have been published. In this investigation, the anti-inflammatory, analgesic effects and representative active ingredient compounds of P. orientale have been studied. Methods: Dried small pieces of the stems and leaves of P. orientale were decocted with water and partitioned successively to obtain ethyl acetate and ethyl ether extract of P. orientale (POEa and POEe). Chemical compositions of them were analyzed by UPLC-Q-Exactive HRMS. Anti-inflammatory and analgesic effects of POEa and POEe were evaluated using xylene induced ear edema, carrageenan induced paw edema, Freunds' complete adjuvant induced arthritis, and formaldehyde induced pain in rat. Their mechanisms of anti-inflammatory and analgesic effects were also studied via assays of TNF-α, IL-1ß, IL-6, and PGE2 in serum. Results: UPLC-Q-Exactive HRMS analysis showed that POEa and POEe mainly contained flavonoids including orientin, isoorientin, vitexin, luteolin, and quercetin. Furthermore, anti-inflammatory effects of POEa and POEe were evident in xylene induced ear edema. The paw edema in Freund's complete adjuvant and carrageenan were significantly (P < 0.05, 0.01) inhibited by POEa (5, 7.5 g/kg). POEe (7.5 g/kg) was significantly (P < 0.05, 0.01) inhibited Freunds' complete adjuvant induced paw edema and cotton pellet induced granuloma formation. Similarly, POEe significantly (P < 0.05, 0.01) inhibited the pain sensation in acetic acid induced writhing test. POEa (5, 7.5 g/kg) significantly (P < 0.05, 0.01) inhibited formaldehyde induced pain in both phases. POEa (7.5 g/kg) markedly (P < 0.05) prolonged the latency period of hot plate test after 30 and 60 min. The concentrations of TNF-α, IL-1ß, IL-6, and PGE2 were significantly (P < 0.01) decreased by POEa (3.75, 5 g/kg). Conclusion: POEa and POEe have anti-inflammatory and analgesic effects, which was mainly relevant to the presence of flavonoids, including orientin, isoorientin, vitexin, luteolin, and quercetin. The mechanism of anti-inflammatory and analgesic effects of POEa may be to decrease the concentrations of TNF-α, IL-1ß, IL-6, and PGE2 in serum.

Feasibility and efficacy of a remote real-time wireless ECG monitoring and stimulation system for management of ventricular arrhythmia in rabbits with myocardial infarction.

The purpose of this study was to explore the feasibility of continuous remote monitoring, and the induction and termination of malignant ventricular arrhythmias (VAs) by a novel implantable electronic cardiovascular device (IECD) system in rabbits with myocardial infarction (MI). The IECD was implanted and MI was induced by ligation of the left anterior descending coronary artery in 20 adult rabbits. Internet-based remote electrocardiogram (ECG) monitoring and ventricular stimulation were conducted in remote locations with internet access. The voltage amplitudes of the stimulation signals were recorded synchronously by remote and surface ECG. Programmed stimulation with regular stimuli and regular stimuli with an added extra stimulus were performed prior to and following the MI surgery to induce and terminate VAs. IECD implantation and MI surgery, as well as qualified remote and bidirectional signal communications between the implanted IECD and extracorporeal system, were successfully achieved in 18 rabbits. The voltage of the stimulation signals recorded by the remote and surface ECGs showed a good correlation with the stimulation current (remote ECG, r=0.972 and surface ECG, r=0.988; P<0.001). Sustained ventricular tachycardia (VT) was induced in five rabbits (5/20, 25%) prior to MI induction and in 12 rabbits (12/16, 75%) following MI induction. Of the 17 induced VTs, 16 were successfully terminated by remote ventricular stimulation. The novel IECD system provides qualified remote wireless ECG monitoring and possesses the potential to induce and terminate VAs by remote ventricular pacing in this rabbit model of MI. Thus, this model of MI may be used to test the efficacy of novel drugs and devices for the management of VAs.