[Investigation of the chemical components of Ciwujia injection using ultra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry].

Ciwujia injection is commonly used to treat cerebrovascular and central nervous system diseases in clinical practice. It can significantly improve blood lipid levels and endothelial cell function in patients with acute cerebral infarction and promote the proliferation of neural stem cells in cerebral ischemic brain tissues. The injection has also been reported to have good curative effects on cerebrovascular diseases, such as hypertension and cerebral infarction. At present, the material basis of Ciwujia injection remains incompletely understood, and only two studies have reported dozens of components, which were determined using high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF MS). Unfortunately, the lack of research on this injection restricts the in-depth study of its therapeutic mechanism.In the present study, a qualitative method based on ultra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry (UHPLC-Q/Orbitrap HRMS) was developed to analyze the chemical components of Ciwujia injection. Separation was performed on a BEH Shield RP18 column (100 mm×2.1 mm, 1.7 µm) using 0.1% formic acid aqueous solution (A) and acetonitrile (B) as the mobile phases, and gradient elution was performed as follows: 0-2 min, 0%B; 2-4 min, 0%B-5%B; 4-15 min, 5%B-20%B; 15-15.1 min, 20%B-90%B; 15.1-17 min, 90%B. The flow rate and column temperature were set to 0.4 mL/min and 30 ℃ respectively. MS1 and MS2 data were acquired in both positive- and negative-ion modes using a mass spectrometer equipped with an HESI source. For data post-processing, a self-built library including component names, molecular formulas, and chemical structures was established by collecting information on the isolated chemical compounds of Acanthopanax senticosus. The chemical components of the injection were identified by comparison with standard compounds or MS2 data in commercial databases or literature based on precise relative molecular mass and fragment ion information. The fragmentation patterns were also considered. For example, the MS2 data of 3-caffeoylquinic acid (chlorogenic acid), 4-caffeoylquinic acid (cryptochlorogenic acid), and 5-caffeoylquinic acid (neochlorogenic acid) were first analyzed. The results indicated that these compounds possessed similar fragmentation behaviors, yielding product ions at m/z 173 and m/z 179 simultaneously. However, the abundance of the product ion at m/z 173 was much higher in 4-caffeoylquinic acid than in 5-caffeoylquinic acid or 3-caffeoylquinic acid, and the fragment signal at m/z 179 was much stronger for 5-caffeoylquinic acid than for 3-caffeoylquinic acid. Four caffeoylquinic acids were identified using a combination of abundance information and retention times. MS2 data in commercial database and literature were also used to identify unknown constituents. For example, compound 88 was successfully identified as possessing a relative molecular mass and neutral losses similar to those of sinapaldehyde using the database, and compound 80 was identified as salvadoraside because its molecular and fragmentation behaviors were consistent with those reported in the literature. A total of 102 constituents, including 62 phenylpropanoids, 23 organic acids, 7 nucleosides, 1 iridoid, and 9 other compounds, were identified. The phenylpropanoids can be further classified as phenylpropionic acids, phenylpropanols, benzenepropanals, coumarins, and lignans. Among the detected compounds, 16 compounds were confirmed using reference compounds and 65 compounds were identified in Ciwujia injection for the first time. This study is the first to report the feasibility of using the UHPLC-Q/Orbitrap HRMS method to quickly and comprehensively analyze the chemical components of Ciwujia injection. The 27 newly discovered phenylpropanoids provide further material basis for the clinical treatment of neurological diseases and new research targets for the in-depth elucidation of the pharmacodynamic mechanism of Ciwujia injection and its related preparations.

[Discovery of exogenous ligands for orphan receptor BRS-3 from Chinese herbs].

Bombesin receptor subtype-3(BRS-3) is an orphan receptor in the bombesin receptor family. Its signal transduction mechanism and biological function have attracted much attention. Seeking the ligand for BRS-3 is of great significance for exploring its function. Considering the fact that the activation of BRS-3 receptor can induce the change in intracellular Ca~(2+) concentration, the fluo-rometric imaging plate reader(FLIPR) was utilized for ligand screening at the cellular level. Among more than 400 monomeric compounds isolated from Chinese herbs, yuanhunine from Corydalis Rhizoma and sophoraisoflavanone A and licoriphenone from Glycyrrhizae Radix et Rhizoma antagonized BRS-3 to varying degrees. It was confirmed in HEK293 cells expressing BRS-3 that yuanhunine, sophoraisoflavanone A, and licoriphenone inhibited the calcium current response after the activation of BRS-3 by [D-Phe~6,ß-Ala~(11),Phe~(13),Nle~(14)]bombesin-(6-14) in a dose-dependent manner with the IC_(50) values being 8.58, 4.10, and 2.04 µmol·L~(-1), respectively. Further study indicated that yuanhunine and sophoraisoflavanone A exhibited good selectivity for BRS-3. In this study, it was found for the first time that monomers derived from Chinese herbs had antagonistic activity against orphan receptor BRS-3, which has provided a tool for further study of BRS-3 and also the potential lead compounds for new drug discovery. At the same time, it provides reference for the research and development of innovative drugs based on the active ingredients of Chinese herbs.

Isoquinoline alkaloid dimers with dopamine D1 receptor activities from Menispermum dauricum DC.

A phytochemical investigation on chemical constituents from the rhizomes of Menispermum dauricum DC. identified eight undescribed dimeric alkaloids with structurally diverse monomeric isoquinoline. Alkaloid structures were elucidated by a combination of spectroscopic data analyses and time-dependent density functional theory (TDDFT) ECD calculation. The isolates were evaluated for inhibitory effect on dopamine D1 receptor and compound 1 exhibited potent D1 receptor antagonistic activity with an IC50 value of 8.4 ± 2.0 µM.

Comprehensive profiling and characterization of the absorbed components and metabolites in mice serum and tissues following oral administration of Qing-Fei-Pai-Du decoction by UHPLC-Q-Exactive-Orbitrap HRMS.

Qing-Fei-Pai-Du decoction (QFPDD) is a Chinese medicine compound formula recommended for combating corona virus disease 2019 (COVID-19) by National Health Commission of the People's Republic of China. The latest clinical study showed that early treatment with QFPDD was associated with favorable outcomes for patient recovery, viral shedding, hospital stay, and course of the disease. However, the effective constituents of QFPDD remain unclear. In this study, an UHPLC-Q-Orbitrap HRMS based method was developed to identify the chemical constituents in QFPDD and the absorbed prototypes as well as the metabolites in mice serum and tissues following oral administration of QFPDD. A total of 405 chemicals, including 40 kinds of alkaloids, 162 kinds of flavonoids, 44 kinds of organic acids, 71 kinds of triterpene saponins and 88 kinds of other compounds in the water extract of QFPDD were tentatively identified via comparison with the retention times and MS/MS spectra of the standards or refereed by literature. With the help of the standards and in vitro metabolites, 195 chemical components (including 104 prototypes and 91 metabolites) were identified in mice serum after oral administration of QFPDD. In addition, 165, 177, 112, 120, 44, 53 constituents were identified in the lung, liver, heart, kidney, brain, and spleen of QFPDD-treated mice, respectively. These findings provided key information and guidance for further investigation on the pharmacologically active substances and clinical applications of QFPDD.

A broad-spectrum substrate for the human UDP-glucuronosyltransferases and its use for investigating glucuronidation inhibitors.

Strong inhibition of the human UDP-glucuronosyltransferase enzymes (UGTs) may lead to undesirable effects, including hyperbilirubinaemia and drug/herb-drug interactions. Currently, there is no good way to examine the inhibitory effects and specificities of compounds toward all the important human UGTs, side-by-side and under identical conditions. Herein, we report a new, broad-spectrum substrate for human UGTs and its uses in screening and characterizing of UGT inhibitors. Following screening a variety of phenolic compound(s), we have found that methylophiopogonanone A (MOA) can be readily O-glucuronidated by all tested human UGTs, including the typical N-glucuronidating enzymes UGT1A4 and UGT2B10. MOA-O-glucuronidation yielded a single mono-O-glucuronide that was biosynthesized and purified for structural characterization and for constructing an LC-UV based MOA-O-glucuronidation activity assay, which was then used for investigating MOA-O-glucuronidation kinetics in recombinant human UGTs. The derived Km values were crucial for selecting the most suitable assay conditions for assessing inhibitory potentials and specificity of test compound(s). Furthermore, the inhibitory effects and specificities of four known UGT inhibitors were reinvestigated by using MOA as the substrate for all tested UGTs. Collectively, MOA is a broad-spectrum substrate for the human UGTs, which offers a new and practical tool for assessing inhibitory effects and specificities of UGT inhibitors.

Inhibition of drug-metabolizing enzymes by Qingfei Paidu decoction: Implication of herb-drug interactions in COVID-19 pharmacotherapy.

Corona Virus Disease 2019 (COVID-19) has spread all over the world and brings significantly negative effects on human health. To fight against COVID-19 in a more efficient way, drug-drug or drug-herb combinations are frequently used in clinical settings. The concomitant use of multiple medications may trigger clinically relevant drug/herb-drug interactions. This study aims to assay the inhibitory potentials of Qingfei Paidu decoction (QPD, a Chinese medicine compound formula recommended for combating COVID-19 in China) against human drug-metabolizing enzymes and to assess the pharmacokinetic interactions in vivo. The results demonstrated that QPD dose-dependently inhibited CYPs1A, 2A6, 2C8, 2C9, 2C19, 2D6 and 2E1 but inhibited CYP3A in a time- and NADPH-dependent manner. In vivo test showed that QPD prolonged the half-life of lopinavir (a CYP3A substrate-drug) by 1.40-fold and increased the AUC of lopinavir by 2.04-fold, when QPD (6 g/kg) was co-administrated with lopinavir (160 mg/kg) to rats. Further investigation revealed that Fructus Aurantii Immaturus (Zhishi) in QPD caused significant loss of CYP3A activity in NADPH-generating system. Collectively, our findings revealed that QPD potently inactivated CYP3A and significantly modulated the pharmacokinetics of CYP3A substrate-drugs, which would be very helpful for the patients and clinicians to avoid potential drug-interaction risks in COVID-19 treatment.

[Investigation of modulating effect of Qingfei Paidu Decoction on host metabolism and gut microbiome in rats].

This study is to explore the effect of Qingfei Paidu Decoction(QPD) on the host metabolism and gut microbiome of rats with metabolomics and 16 S rDNA sequencing. Based on 16 S rDNA sequencing of gut microbiome and metabolomics(GC-MS and LC-MS/MS), we systematically studied the serum metabolites profile and gut microbiota composition of rats treated with QPD for continued 5 days by oral gavage. A total of 23 and 43 differential metabolites were identified based on QPD with GC-MS and LC-MS/MS, respectively. The involved metabolic pathways of these differential metabolites included glycerophospholipid metabolism, linoleic acid metabolism, TCA cycle and pyruvate metabolism. Meanwhile, we found that QPD significantly regulated the composition of gut microbiota in rats, such as enriched Romboutsia, Turicibacter, and Clostridium_sensu_stricto_1, and decreased norank_f_Lachnospiraceae. Our current study indicated that short-term intervention of QPD could significantly regulate the host metabolism and gut microbiota composition of rats dose-dependently, suggesting that the clinical efficacy of QPD may be related with the regulation on host metabolism and gut microbiome.

Renal abscess in children: a 10-year clinical and radiologic experience in a tertiary medical center.

Renal abscess is a rare disease in childhood and early diagnosis is essential to minimize the renal damages. Experiences gained from this 10-year study on patients with CT-proved diagnosis include the varied clinical presentations and use of sonographic-marked nephromegaly and focal mass as indications for further CT diagnosis.

An unusual presentation of large pneumoencephaly after frequent falls in an elderly patient.

Pneumoencephaly is usually a complication of head trauma or craniotomy. We present a patient with large pneumoencephaly with acute neurological deficit after frequent falls, but without recent major trauma or surgery. Skull X-ray films show intracranial air in the frontal region and in the 2nd, 3rd, and 4th ventricles, reminiscent of an pneumoencephalogram. Computed tomography (CT) of the head shows air in the right frontal region and lateral ventricles. After craniotomy to relieve the pressure of the air, the clinical condition of the patient improved significantly after six months.

[Effects of centipede extract on life span of drosophila].

OBJECTIVE: To evaluate effects of centipede on life span of drosophila and on the appearance of the filial generation. METHODS: Drosophila melanogaster (male and female) were cultured in tubes with centipede extraction at concentrations of 0, 0.2%, 1%, 2.5%, or 5% until all drosophila died. The appearance of the filial generation were observed. RESULTS: The life span, including average, longest and shortest life span was shortened in exposed groups of both sexes and significantly dose-dependent. The appearance of the filial generation showed normal. CONCLUSION: Centipede extracts accelerates the aging of drosophila and shortens their life span. The appearance of the filial generation is not affected. Clinic using centipede for a long period should be carefully.

Early administration of hyperbaric oxygen therapy in distraction osteogenesis--a quantitative study in New Zealand rabbits.

BACKGROUND: We investigated the effect of hyperbaric oxygen (HBO) therapy on the early phase of tibial lengthening in our established rabbit model. METHODS: Twenty-four male rabbits (six per group) underwent right tibial lengthening by 5 mm. Group 1 then underwent 2.5 atmospheres of absolute hyperbaric oxygenation for 2 hours daily for 6 weeks postoperatively; group 2, for early 5 weeks (weeks 1-5), group 3, for late 5 weeks (weeks 2-6), and group 4 had no HBO therapy. Bone mineral density (BMD) was measured before surgery and weekly thereafter from weeks 2 through 6. The mechanical strengths of the lengthened tibias were measured. RESULTS: Significantly higher mean %BMDs were obtained for groups 1 and 2 compared with groups 3 and 4. There was no difference in the mean %BMD between groups 1 and 2 (p > 0.05). The results were similar for mean percentage maximal torque; group 1 had the maximum torque, followed sequentially by groups 2 though 4. CONCLUSION: The study results suggest that early and full-term administration of HBO therapy on tibial lengthening may achieve better benefits.

Connatal tuberculosis in a very premature infant.

UNLABELLED: Connatal tuberculosis is increasing in incidence and the mortality and morbidity of this disease remains high. We report a 27-week-old, 896 g female premature infant who had mild respiratory distress syndrome after birth. She developed signs of infection, progressive pneumonia and atelectasis which did not respond to mechanical ventilation and antibiotics. At 41 days of age, Mycobacterium tuberculosis was isolated from the non-bronchoscopic bronchoalveolar lavage. The isolate was sensitive to isoniazid, rifampin, streptomycin, and pyrazinamide. Miliary tuberculosis was subsequently diagnosed in her mother on a chest X-ray film and sputum cultures. The infant was treated successfully with anti-tuberculosis drugs. She had normal growth and development at the chronological age of 20 months old. CONCLUSION: Connatal tuberculosis should be considered in premature infants with symptoms of sepsis refractory to antibiotics. Most premature infants with connatal tuberculosis have lung involvement, and non-bronchoscopic bronchoalveolar lavage can be a useful procedure to establish the diagnosis.

Extravasation of calcium gluconate concomitant with osteomyelitis in a neonate.

Extravasation of calcium gluconate is not an uncommon complication in neonatal units, which may result in skin and soft tissue inflammation, injury and even infection. We report an 11-day-old preterm infant who developed osteomyelitis at an infiltrated intravenous site through which calcium gluconate was infused. The patient was initially treated conservatively as a simple calcium-infiltrate. Clinical condition deteriorated until proper antibiotic treatment was given. We exclude other possible causes and assume that calcium-infiltrate had an important role in the pathogenesis. We conclude that osteomyelitis should be considered in a patient with calcium extravasation whose soft tissue inflammation does not improve as expected. Furthermore, using a diluted calcium solution and checking the insertion site frequently in order to identify extravasation earlier would prevent the complication.

Thoracic wall lesions in children.

Our objective was to review the clinical features and radiographic manifestations of chest wall lesions in a pediatric chest unit. Twenty-five patients (11 males and 14 females, aged 14 months to 15 years) were suspected of having a chest wall lesion on the basis of clinical examination and chest radiograph. A retrospective review of clinical characteristics and radiographic findings was performed in order to define the pathology and extent of lesions and outcomes of chest wall diseases in children. Thirty-six percent of pediatric patients studied for thoracic abnormalities had anatomical variations of the thoracic cage such as sternal titling, bifid ribs, and costal cartilage convexities. Tuberculous infections of the sternum and ribs were seen in 4 cases. Osteochondromas were seen in 4 patients, and were especially noted in patients with a history of familial exostosis. Other malignant thoracic wall tumors included in this study included chondrosarcoma (n=1), Ewing's sarcoma (n=1), and primitive neuroectodermal tumor (n=2). In conclusion, the protruding lesions arising from the chest wall demonstrate a wide range of normal, congenital variant, and pathologic lesions arising from the soft tissues and bony components. Those with life-threatening lesions (less likely located in the parasternal region) are more prone to have chest pains and dyspneic respirations; in most cases, plain chest radiographs tend to have a high rate of bony or pleural involvement as well as thoracic deformities. In an endemic area of tuberculosis, costal tuberculosis should be considered in undiagnosed bony lesions; a chest CT scan could be diagnostically useful.

Use of 3-dimensional computed tomography scan to evaluate upper airway patency for patients undergoing sleep-disordered breathing surgery.

OBJECTIVE: Our goal was to investigate the correlation between upper airway 3-dimensional computed tomography (3-D CT) measurements and polysomnography data of patients with sleep-disordered breathing (SDB). Study design and setting The 3-D CT scan measurements of the retropalatal space (RP), lateral and anteroposterior diameters of RP, retroglossal space, uvular thickness, and volume of upper airway were taken prospectively from 194 consecutive SDB patients. These were correlated with patients' respective Respiratory Distress Index (RDI), snoring index, and lowest arterial oxygen saturation gathered from polysomnographic data. RESULTS: RP (r = -0.27, P = 0.0001) and lateral diameter (P = 0.0073) were significantly correlated with RDI. Multivariate regression model (R(2) = 0.286) demonstrates RP (P = 0.0036) and retroglossal space (P = 0.027) to be predictive of RDI. No 3-D CT parameters correlated with snoring index and lowest arterial oxygen saturation attained statistical significant. CONCLUSION: The 3-D CT scan measurements can be valuable in evaluating upper airway patency in patients with SDB. The RP was the most relevant upper airway anatomic structure identified in these patients. SIGNIFICANCE: Modification of palatal surgery to increase the lateral dimension of the RP may be a breakthrough in increasing its success rate.

Catheter-related superior vena cava syndrome complicated by chylothorax in a premature infant.

A premature infant experienced respiratory distress due to chylothorax. Contrast-enhanced chest computed tomography showed superior vena cava (SVC) obstruction with collaterals, and surgery confirmed an obstruction of the SVC by intravascular fibrotic tissue and thrombi caused by placement of a central venous catheter. The respiratory distress improved after surgical intervention. In the differential diagnosis of acute respiratory distress, it is necessary to consider catheter-related chylothorax and SVC syndrome when a previously stable patient has an acute change in cardiopulmonary status, regardless of the duration of catheter use. Although often considered safe and easy, central venous catheter placement may result in complications. Using appropriate catheters and choosing proper insertion sites can minimize these events. Early diagnosis and treatment can be life-saving should complications occur.

Evaluation of oesophageal atresia without fistula by three-dimensional computed tomography.

UNLABELLED: During the last decade, an increasing number of paediatric surgeons have chosen to perform a delayed oesophageal anastomosis for oesophageal atresia (OA) without tracheo-oesophageal fistula (TOF). The gap between the two oesophageal pouches is an important determinant in the surgical management of these patients. We describe a new method using spiral computed tomography (CT) to evaluate the gap between the proximal and distal oesophageal pouches. In our last four cases of OA without TOF, Stamm gastrostomy was performed soon after birth. At about 4 weeks of age, these patients underwent spiral CT with air inflation via the gastrostomy feeding tube with 3 mm slice thickness, pitch 1, and reconstruction interval every 1.5 mm. A three-dimensional reconstruction was done and measurements of the gap between the two segments were recorded. The procedure was repeated at monthly intervals until the optimal conditions for surgery were observed. All the babies thrived well on gastrostomy feeding. The gap distances were 2.4, 3.8, 4.2 and 5.9 cm respectively. Delayed primary oesophageal anastomosis was accomplished in three cases and gastric transposition was performed in another case at 12, 13, 14 and 16 weeks of age. The exact anatomical position the operation correlated well with the pre-operative three-dimensional CT results. CONCLUSION: this new technique is an easy and non-invasive method to assess the long gap in babies with oesophageal atresia without tracheo-oesophageal fistula. The radiological findings are also consistent with surgical anatomical situation during surgery suggesting that this investigation is useful in the pre-operative planning of oesophageal anastomosis or replacement.

Congenital oligodendroglioma with initial manifestation of jaundice.

Neonatal brain tumor is a rare clinical entity. The initial presentation in affected newborn infants is often subtle, nonspecific, and rarely includes focal neurologic signs. We report a 2-day-old male infant with congenital oligodendroglioma with initial manifestations of hyperbilirubinemia. Phototherapy was started immediately after admission. Because of a tense anterior fontanel and irritable crying, head ultrasonography, computed tomography, and magnetic resonance imaging were performed and revealed a heterogenous brain tumor in the left temporoparietal lobe. Craniotomy and tumor evacuation was performed, and WHO grade III anaplastic oligodendroglioma was confirmed by the pathologic studies. The patient was discharged 3 weeks later, and outpatient follow-up examination revealed a normally developed infant at 6 months of age. The cause of jaundice was thought to be secondary to tumor hemorrhage, which was not infrequent in neonatal brain tumor.