ABSTRACT
Two methods including gas chromatography tandem mass spectrometry (GC-MS/MS) and high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) were established to detect common alkyl sulfonates and aryl sulfonates genotoxic impurities. Four alkyl sulfonates and methyl benzenesulfonate were determined by GC-MS/MS using butyl methanesulfonate as the internal standard, the chromatographic column was HP-5MS UI (30 mm × 0.25 mm, 0.25 µm), the carrier gas was helium, the flow rate was 1.0 mL·min-1 in a constant flow mode, the sample inlet temperature was set to 250 ℃, the split ratio was 10∶1, and the initial temperature of the heating program was 80 ℃, maintained for 1 minute, and then increased to 240 ℃ at a heating rate of 30 ℃·min-1 for 2 minutes. The mass spectrometry detector was an electron bombardment ion source (EI source), the data collection condition was multi reaction monitoring mode (MRM), and method validation using the raw material of clinical drug citalopram hydrobromide as a sample. The results showed that the linear range of four alkyl sulfonates and methyl benzenesulfonate were good at 3-50 ng·mL-1 and 9-150 ng·mL-1, with a correlation coefficient of r > 0.999, The spiked recovery was 80%-120%. The detection limits were 1 and 3 ng·mL-1; Ten aryl sulfonates determined by LC-MS/MS, the chromatographic column was CSH Fluoro phenyl (100 mm × 2.1 mm, 1.7 µm), the mobile phase was methanol (B)-5 mmol·L-1 ammonium formate (D), with a flow rate of 0.2 mL·min-1, and gradient elution was performed. The gradient program (T/% B) was set as 0/20, 25/90, 35/90, 42/20. The mass spectrometer detector was electro spray ionization with positive ionization mode (ESI+), the data collection was in dynamic multi reaction monitoring mode (dMRM), and the method was validated using the raw material of the clinical drug citalopram hydrobromide as a sample. The results showed that the linear range of aryl sulfonates were good at 9-2 000 ng·mL-1, 3-100 ng·mL-1 and 0.9-30 ng·mL-1, respectively. The correlation coefficient r > 0.999, the spiked recovery was 80%-120%. The detection limits were 30, 1 and 0.3 ng·mL-1. Two detection methods did not detect potential sulfonate genotoxicity impurities in the above APIs. The established analytical methods are reliable and effective, which can provide reference for drug quality control and detection.
ABSTRACT
Objective: To analyse the clinical history, laboratory tests and pathological data of a patient who suffered from novel coronavirus pneumonia(COVID-19) and provide reference for the clinical treatment of similar cases. Methods: Data of clinical manifestation, laboratory examination, bronchoscopy, echocardiography and cardiopulmonary pathological results were retrospectively reviewed in a case of COVID-19 with rapid exacerbation from mild to critical condition. Results: This patient hospitalized at day 9 post 2019 novel coronavirus(2019-nCoV) infection, experienced progressive deterioration from mild to severe at day 12, severe to critical at day 18 and underwent extracorporeal membrane oxygenation(ECMO) and continuous renal replacement therapy(CRRT) as well as heart lung transplantation during day 28-45 post infection, and died at the second day post heart and lung transplantation. The patient had suffered from hypertension for 8 years. At the early stage of the disease, his symptoms were mild and the inflammatory indices increased and the lymphocyte count decreased continuously. The patient's condition exacerbated rapidly with multi-organ infections, and eventually developed pulmonary hemorrhage and consolidation, pulmonary hypertension, right heart failure, malignant ventricular arrhythmias, liver dysfunction, etc. His clinical manifestations could not be improved despite viral RNAs test results became negative. The patient underwent lung and heart transplantation and finally died of multi organ failure at the second day post lung and heart transplantation. Pathological examination indicated massive mucus, dark red secretions and blood clots in bronchus. The pathological changes were mainly diffused pulmonary hemorrhagic injuries and necrosis, fibrosis, small vessel disease with cardiac edema and lymphocyte infiltration. Conclusions: The clinical course of severe COVID-19 can exacerbate rapidly from mild to critical with lung, liver and heart injuries.
Subject(s)
Humans , Betacoronavirus , COVID-19 , Coronavirus Infections/pathology , Fatal Outcome , Hemorrhage/virology , Lung/pathology , Myocardium/pathology , Pandemics , Pneumonia, Viral/pathology , Retrospective Studies , SARS-CoV-2ABSTRACT
A new napthalenone, rumexone A (1), was isolated from the roots of Rumex nepalensis. The structure of 1 was elucidated by extensive spectroscopic analyses, including 1D and 2D NMR spectra and MS data. Its cytotoxic effect was evaluated using four clinically relevant human cancer cell lines, gastric carcinoma SGC7901, breast carcinoma MDA-MB-231, lung carcinoma A549, and hepatocellular carcinoma HepG2.
ABSTRACT
BACKGROUND: In the existing thoracic pedicle screw fixation technique, superior facet or combined with transverse process and the imaginary transverse and sagittal section are used as the reference mark of pedicle screw entry point and angle, respectively, which still remain controversial. Because of the effect of position deviation and subjective judgment during surgery, screw misplacement and severe complications often occur, which limit the clinical application and promotion of the technique. At present, it is very important to select a single bony mark, which is not affected by the patient's position, and is easy to judge, as the reference mark of screw entry point and angle. OBJECTIVE: To explore the feasibility of taking superior facet as a single bony mark to determine thoracic pedicle screw parameters in normal adults by measuring 3D reconstruction CT image and it's corresponding relationships. METHODS: 3D reconstruction CT images of the thoracic spine in 30 normal adults were selected and the basic parameters, entry angle parameters and entry depth parameters were directly determined on specific reconstructed CT images of T1-T12.The basic parameters included the vertical distance from the entry point to the midline, the vertical distance from the outer edge of superior facet to the midline and superior facet base width. Entry point parameters were indirectly calculated by the basic parameters, including transversal point-facet distance, entry point ratio and sagittal point-facet distance, which was the horizontal distance from entry point to outer edge of superior facet, the ratio of transversal point-facet distance to superior facet base width, and the longitudinal distance from entry point to the base of superior facet. Entry angle parameters included transversal axis-facet angle and sagittal axis-facet angle, which is the transversal and sagittal angle between the axis of pedicle and the surface of facet. Entry depth parameter included safety screw length, which is the distance from entry point to anterior vertebral cortex along the axis of pedicle. Mean and standard deviation were counted and statistics difference was compared. RESULTS AND CONCLUSION: (1) Using CT technology of 3D reconstruction, T1-T12thoracic transversal section through bilateral pedicle axis, transversal section through the base of the superior facet, which parallel to the bilateral pedicle axis, and oblique sagittal section through homolateral pedicle axis were successfully obtained. (2) The statistical results of the basic parameters were as follows: the left and right vertical distances between T1-T12from the entry point to the midline were (14.6±2.6) and (14.5±2.5) mm, the vertical distance from the outer edge of superior facet to the midline were (15.7±1.9) and (15.7±2.0) mm, and superior facet base width were (8.8±1.3) mm and (8.8±1.1) mm, respectively. There was no significant difference in the measured values between two sides of the above parameters (P=0.343, 0.214, 0.467). (3) The statistical results of pedicle screw parameters were as follows: the left and right transversal point-facet distance between T1-T12were (1.2±1.3) and (1.3±1.4) mm, entry point ratio were (14.3±17.0)% and (13.6±16.1)%, sagittal point-facet distance were (3.4±0.8) and (3.3±0.9) mm, transversal axis-facet angle were (92.4±4.7)° and (92.6±5.0)°, sagittal axis-facet angle were (91.8±4.1)° and (91.7±3.6)°, and safe screw length were (40.7±4.8) and (40.4±4.6) mm, respectively. There was no significant difference in the measured values between two sides of the above parameters (P=0.073, 0.084, 0.310, 0.265, 0.241, 0.175). (4) These results indicate that taking thoracic superior facet as a single anatomic landmark to determine pedicle screw parameters is simple and feasible in normal adults, and they correlate with each other, which can be used as a new method for choosing screw parameters.
ABSTRACT
The chemical constituents of Herpetospermum caudigerum were investigated using chromatographic methods, including silica gel column chromatography, Sephadex LH-20 and semi-preparative HPLC. Four compounds were isolated and their structures were elucidated by spectral data and physicochemical properties, which were identified as 2,11-dimethoxy-3,9-dihydroxy-7H-dibenzo[c,e]oxepin-5-one (1), 7,8'-didehydroherpetotriol (2), herpetotriol (3) and kaempferitrin (4). Among those, compound 1 is one new 7H-dibenzo[c,e] oxepin-5-one, named as herpetolide C.
ABSTRACT
OBJECTIVE: To prepare and evaluate the novel core-shell structural phospholipid-functionalized mesoporous silica nanoparticles (MSN-LP) modified with angiopep-2 (ANG-MSN-LP). METHODS: Mesoporous silica nanoparticles (MSN) was synthesized by the modified Stober method. MSN-PTX was prepared by saturated solution adsorption method. ANG-MSN-LP was developed by selfassembly and film hydration method. By using dialysis bag method to investigate the in vitro drug release characteristics and MTT method to investigate the cytotoxicity on HBMEC and C6 cells. The transport ability and effects on cell cycle of the carrier was investigated by the BBB monolayer model. RESULTS: MSN was synthesized with high specific surface area (SBET, 425 m · g-1), cumulative pore volume (Vp, 0.37 cm · g-1) and pore size(3.5 nm). PTX was highly encapsulated (drug loading efficiency up to 11.1%) into MSN. Results of in vitro release showed that about 75.5% of PTX released from ANG-MSN-LP-PTX after 48 h and burst release was effectively reduced compared with MSN-PTX or PTX solution, indicating pronounced sustained-release characteristics. The good biocompatibility and low toxicity of ANG-MSN-LP were evaluated by HBMEC and C6 cells. The transport ratio was 2.49% for PTX, 2.72% for MSN-PTX, 4.45% for MSN-LP-PTX and 10.74% for ANG-MSN-LP-PTX respectively. In addition, ANG-MSN-LP-PTX showed a higher cell number of G2-M phase of 40.92 ± 6.20%. CONCLUSION: ANG-MSN-LP is a prospective targeting drug delivery system for therapy of brain glioma. Meanwhile, saturated solution adsorption method can increase the drug loading efficiency highly.
ABSTRACT
The purpose of this study was to investigate the absorption kinetics of aconitine, mesaconitine and hypaconitine in rats after oral administration of Sini Tang powder. With cannulate portal and jugular veins cannulated (double-cannulate), conscious moving rats were orally administered Sini Tang. Then samples of portal and systemic blood were collected at the designated periods of time and analyzed for aconitine, mesaconitine and hypaconitine by HPLC. Apparent absorption coefficient of aconitine, mesaconitine and hypaconitine was caculated respectively. The results indicated that the apparent absorption coefficient of aconitine, mesaconitine and hypaconitine come from Sini Tang were 0. 336, 0. 090, 0. 176, respectively, which had some differences among them. It was also suggested that double-cannulated rat was useful for estimating the absorption kinetics of aconitine, mesaconitine and hypaconitine after orally administered Sini Tang by determining the AUC values for drugs in portal and systemic blood samples. The three alkaloids could all be detected in blood, but the absorption differences were existed among the three alkaloids.
Subject(s)
Animals , Female , Male , Rats , Absorption , Administration, Oral , Alkaloids , Blood , Pharmacokinetics , Area Under Curve , Diterpenes , Blood , Pharmacokinetics , Drugs, Chinese Herbal , Pharmacokinetics , Kinetics , Powders , Pharmacokinetics , Rats, Sprague-DawleyABSTRACT
Objective To clone, express and purify human PIF1 protein and analyze its ATPase activity. Methods The PIF1 cDNA was amplified by PCR from HeLa cell cDNA library and inserted to pET24b with histidine tag at its terminus to form pET24b-PIF1 plasmid. The recombinant pET24b-PIF1 plasmid was transformed to RosettaTM 2 (DE3) and the expression of PIF1 protein was monitored by SDS-PAGE analysis. By using fast protein liquid chromatograph (FPLC) system, the PIF1 protein was purified by affinity chromatograph and gel filtration. The ATPase activity of PIF1 was checked by thin layer chromatograph(TLC). Results The PIF1 protein was successfully cloned and expressed in E.coli. Conclusions The purification procedure of PIF1 protein was established using FPLC. The overexpressed and the purified PIF1 helicase has DNA and Mg2+ dependent ATPase activity.
ABSTRACT
<p><b>OBJECTIVE</b>To study on the relationship of serum vitamin E and liver pathological features in the patients with chronic hepatitis B.</p><p><b>METHODS</b>Sixty-six patients with chronic hepatitis B and ten healthy controls were enrolled in this present study. The serum vitamin E level was measured spectrophotometrically. Comparisons of liver function test, HBeAg and HBV DNA level were conducted among different liver pathological features including inflammatory grading and fibrosis staging.</p><p><b>RESULTS</b>Compared with healthy controls, the serum level of vitamin E was significantly decreased in the patients with chronic hepatitis B, especially in those with elevated ALT activity. In comparison between HBeAg positive group and HBeAg negative group, the serum level of vitamin E of the former group did not significantly changed (P > 0.05). Furthermore, the serum level of vitamin E has been demonstrated to be negatively associated with the inflammation grading in the patients with chronic hepatitis B. However, there was no significant association between the serum vitamin E and liver fibrosis staging.</p><p><b>CONCLUSION</b>Vitamin E, as one of the important anti-oxidants, was demonstrated to be implicated in the progression of liver inflammation in the patients with chronic hepatitis B. Furthermore, the supplement of vitamin E would be a potential therapy for attenuate the inflammatory response.</p>