Humans ; Consensus ; Pancreatic Neoplasms/therapy* ; Neuroendocrine Tumors/therapy*

Aim To investigate the effect of lactate dehydrogenase inhibitor on LPS/D-Gal-induced acute liver injury in mice. Methods BALB/ C mice were divided into four groups:solvent control group, lactate dehydrogenase inhibitor NHI-2 group, lipopolysaccharide(LPS)/ D-galactosamine(D-Gal)group and LPS/D-Gal+NHI-2 group. To induce acute liver injury, mice were injected intraperitoneally with LPS(10 μg·kg-1)and D-Gal(700 mg·kg-1), NHI-2 was intraperitoneally injected 30 min before LPS/D-Gal exposure. Liver tissue and serum were harvested 1.5 or 6 h after LPS/D-Gal exposure, serum lactate, serum aspartate aminotransferase(ALT), serum alanine aminotransferase(AST), serum tumor necrosis factor alpha(TNF-α)liver malondialdehyde(MDA)and liver caspase-3/8/9 levels were determined. HE staining was used to evaluate the degree of liver injury. TUNEL staining was used to evaluate hepatocyte apoptosis. Survival curve was used to record survival situation of tested mice. Results Serum lactate level of model mice was significantly reduced after treatment with NHI-2. Compared with LPS/D-Gal group, level of serum TNF-α showed no significant difference, but serum ALT and AST level of LPS/D-Gal+NHI-2 group significantly decreased, injury of liver structure was remarkably attenuated, level of MDA and activity of caspase-3/8/9 in liver were significantly down-regulated, and the number of TUNEL-positive cells was significantly reduced. Treatment with NHI-2 also significantly improved the survival rate of LPS/D-Gal-insulted mice. Conclusion Lactate dehydrogenase inhibitor alleviates LPS/D-Gal-induced acute liver injury in mice.

Scaffold hopping refers to computer-aided screening for active compounds with different structures against the same receptor to enrich privileged scaffolds, which is a topic of high interest in organic and medicinal chemistry. However, most approaches cannot efficiently predict the potency level of candidates after scaffold hopping. Herein, we identified potent PDE5 inhibitors with a novel scaffold via a free energy perturbation (FEP)-guided scaffold-hopping strategy, and FEP shows great advantages to precisely predict the theoretical binding potencies ΔG _FEP between ligands and their target, which were more consistent with the experimental binding potencies ΔG _EXP (the mean absolute deviations | Δ G FEP - Δ G EXP |  < 2 kcal/mol) than those ΔG _MM-PBSA or ΔG _MM-GBSA predicted by the MM-PBSA or MM-GBSA method. Lead L12 had an IC₅₀ of 8.7 nmol/L and exhibited a different binding pattern in its crystal structure with PDE5 from the famous starting drug tadalafil. Our work provides the first report via the FEP-guided scaffold hopping strategy for potent inhibitor discovery with a novel scaffold, implying that it will have a variety of future applications in rational molecular design and drug discovery.

Eight sesquiterpenoids were isolated from petroleum ether extract of Aquilariae Lignum Resinatum by various column chromatography techniques including silica gel, ODS, and semi-preparative HPLC. Their structures were identified on the basis of physicochemical properties, UV, IR, MS, and NMR spectroscopic data as(4S,5S,7R,10S)-5,7-dihydroxy-11-en-eudesmane(1),(7R,10S)-eudesma-4-en-11,15-diol(2),(2R,4S,5R,7R)-2-hydroxyeremophila-9,11-dien-8-one(3), 7α-H-9(10)-ene-11,12-epoxy-8-oxoeremophilane(4),(+)-9β,10β-epoxyeremophila-11(13)-en(5), 4(14)-eudesmene-8α,11-diol(6), 12,15-dioxo-selina-4,11-dien(7), and 2β,8 aα-dihydroxy-11-en-eremophilane(8). Compounds 1 and 2 are new compounds, and their absolute configurations were determined by calculating ECD. Compounds 1, 4, and 6-8 could significantly improve taurocholic acid(TCA)-induced gastric mucosal GES-1 cell injury at a concentration of 20 μmol·L~(-1), and the cell protection rates were 23.51%±2.79%, 16.10%±1.25%, 24.45%±4.89%, 17.48%±2.93%, and 21.44%±2.39%, respectively.
Chromatography, High Pressure Liquid ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Sesquiterpenes/chemistry*

Due to the limited resource of bear bile powder, the major raw material of Tanreqing Capsules(TRQ), cultured bear bile powder is used as a replacement to develop the Tanreqing Capsules Substitute(TRQS). An LC-MS/MS method was established in this study for simultaneous quantitation of 8 compounds from TRQS in rat plasma: tauroursodeoxycholic acid(TUDCA), taurocheno-deoxycholic acid(TCDCA), ursodeoxycholic acid(UDCA), chenodeoxycholic acid(CDCA), ferulic acid, wogonoside, baicalin, and forsythoside A. Thereby, the pharmacokinetic behaviors of TRQ and TRQS were evaluated. Concentration of endogenous compounds TUDCA, TCDCA, UDCA, and CDCA was determined with the stable isotope surrogate analytes: D4-TUDCA, D4-TCDCA, D4-UDCA, and D4-CDCA. Plasma samples were extracted by acetonitrile-induced protein precipitation. The LC conditions are as follows: Waters BEH C_(18) column(2.1 mm×100 mm, 1.7 μm), mobile phase of 10 mmol·L~(-1) ammonium formate aqueous solution(containing 0.01% formic acid) and acetonitrile-methanol mixture(1∶5). MS conditions are as below: multiple reaction monitoring(MRM), ESI~(+/-). Concentration of UDCA, CDCA, TUDCA, and TCDCA was corrected with a response factor, which is the ratio between the responses recorded for the surrogate and the authentic analyte at the equal concentration. Each of the plasma components showed good linearity(r > 0.995 1). Accuracy and precision met the criteria(inter-day RSD<7.0%, RE 89.98%-112.0%; intra-day RSD<12%, RE 90.41%-111.2%). The recovery was 64.83%-119.9% and matrix effect was 87.15%-113.8%. The validated method was applied for pharmacokinetic study of TRQS and TRQ(po, 0.94 g·kg~(-1)). There was no significant difference in C_(max) and AUC_(0-24 h) of baicalin, UDCA, TUDCA, and TCDCA between the two groups, indicating similar pharmacokinetic behaviors between TRQS and TRQ in rats.
Animals ; Capsules ; Chromatography, Liquid ; Drugs, Chinese Herbal/pharmacokinetics* ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Tandem Mass Spectrometry

OBJECTIVE: To investigate the clinical significance of the targeted next-generation sequencing assay for patients with suspected myeloid malignancies. METHODS: A total of 39 hematopenia patients with suspected myeloid malignamies in Department of Hematology of The Affiliated Huai'an No.1 People's Hospital of Nanjing Medical University from January 2018 to April 2019 were treated, 20 hot spot genes of myelodysplastic syndrome (MDS) were detected. RESULTS: Regarding the diagnostic type, there were 7 cases of idiopathic cytopenia of undetermined significance (ICUS), 8 cases of clonal cytopenias of undetermined significance (CCUS) and 24 cases of myeloid myeloid malignancies which included 18 cases of MDS, 4 cases of myelodysplastic/myeloproliferative neoplasms (MDS/MPN) and 2 cases of acute myeloid leukemia. Positive mutation was detected in 70.8% (17/24) of myeloid malignancy patients , and 72.7% (16/22) in MDS and MDS/MPN patients. The main mutation types were ASXL1, TET2 and RUNX1. Compared with gene negative group, there were no significant differences in sex, age (＜60 years old or ≥60 years old), proportion of bone marrow blast cells (＜5% or≥5%) and cytogenetics (good, medium and poor) (P＞0.05). Furthermore, all 8 CCUS patients showed positive mutation, and the incidence of double or multiple mutation in CCUS group was significantly lower than that of the MDS and MDS/MPN group (37.5% vs 54.5%) (P=0.002). The mutation types between the two groups were similar, and there was no significant difference in variant allele frequency (P＞0.05). CONCLUSION Our results suggest that there are high rates of double or multiple mutations in myeloid malignancies, especially in patients with MDS and MDS/MPN. Targeted sequencing assay can improve the diagnosis of myeloid malignancies, and guide clinical treatment.
Humans ; Leukemia, Myeloid, Acute/genetics* ; Middle Aged ; Mutation ; Myelodysplastic Syndromes/genetics* ; Myelodysplastic-Myeloproliferative Diseases ; Patients

Objective:To investigate the topological alterations in brain functional networks following comprehensive treatment including brain-computer interface (BCI) training in subacute stroke subjects. Methods:From January, 2018 to June, 2019, 14 subacute stroke patients with moderate to severe upper limbs paralysis accepted routine physical therapy, occupational therapy and BCI training based on motor imagery, for four weeks. They were assessed with Fugl-Meyer Assessment-Upper Extremities (FMA-UE), Action Research Arm Test (ARAT) and Wolf Motor Function Test (WMFT) before and after treatment, while the functional connectivity (FC) was investigated with resting state functional magnetic resonance imaging. Results:The scores of FMA-UE, ARAT and WMFT increased after treatment (|t| > 5.298, Z = -3.297, P < 0.01). The FC also increased across the whole brain, including temporal, parietal, occipital lobes and subcortical regions. The FC between left piriform cortex of parietal lobule (BA5L) and right medial surface of temporal lobe (BA48R), as well as those between left precentral gyrus (BA4L) and right anterior transverse temporal gyrus (BA41R) (r > 0.416, P < 0.05). Conclusion:Comprehensive rehabilitation including BCI training may promote recovery of motor function and activities of FC in brain in subacute stroke patients.

Objective: To investigate the effect of diabetes on the risk of stroke recurrence within 1 year after onset of ischemic stroke. Methods: Patients with ischemic stroke admitted to the Department of Neurology, the Second Hospital of Hebei Medical University from October 2016 to August 2017 were enrolled prospectively. Their baseline clinical data were collected and they were followed up for one year. The risk factors for ischemic stroke in recurrence group and non-recurrence group were compared. Cox proportional hazard regression model was used to determine the independent risk factors for ischemic stroke recurrence. Kaplan-Meier survival analysis was used to explore the impact of risk factors on the risk of stroke recurrence. Results: A total of 1 436 patients with ischemic stroke were included. During the follow-up of 1 year, a total of 183 patients had recurrence (12.74%). There were significant differences in the proportion of patients with diabetes, atrial fibrillation, hyperhomocysteinemia, oral antiplatelet drugs, and statins after discharge, and baseline fasting blood glucose level between the recurrence and the non-recurrence group (all P<0.05). Multivariate Cox proportional hazards analysis showed that diabetes mellitus (hazard ratio [HR] 1.574, 95% confidence interval [CI] 1.161-2.134; P=0.003) was an independent risk factor for stroke recurrence. Taking statins (HR 0.686, 95% CI 0.481-0.979; P=0.038) and antiplatelet agents (HR 0.678, 95% CI 0.467-0.983; P=0.041) after discharge were the independent protective factors for ischemic stroke recurrence. Kaplan-Meier survival analysis showed that the recurrence rate of ischemic stroke in patients with diabetes was significantly higher than that in those without diabetes (log-rank test, P=0.003). The recurrence rate of stroke in patients taking statins and antiplatelet drugs was significantly lower than that in patients who did not take the corresponding drugs (log-rank test, all P<0.001). Conclusions Diabetes is an independent risk factor for ischemic stroke recurrence, and taking statins and antiplatelet drugs are the independent protective factors for ischemic stroke recurrence.

The present study is to develop an HPLC-ELSD method for simultaneous determination of three pairs of triterpenoid isomers, Ilexsaponin A₁, Ilexhainanoside D, Ilexgenin A, 3β, 19α-dihydroxyolean-12-ene-24, 28-dioic acid (ilexhainanin E) ursolic acid and oleanic acid in the leaf of Ilex hainanensis, which could provide evidence to the quality control of this herb. The six constituents were measured on a Waters XBridge C₁₈ column (4.6 mm×250 mm, 5 μm), with a mobile phase consisting of methanol (A)- 0.5% formic acid in water (B) at a flow rate of 1.0 mL·min⁻¹ (0-18 min，70%-85% A，18-20 min，85%-95% A；20-35 min，95% A). The carrier gas was N₂, and the pressure was 2.8 L·min⁻¹. The drift tube in this experiment were set at 70 °C. The injection volume was 10 μL. The contents of the six triterpenoids in 6 samples were 3.7-8.5, 10.3 -22.1, 2.8-5.9, 7.8-14.1, 2.6-3.8 and 8.8-11.9 mg·g⁻¹, respectively. The established method is proved to be accurate and sensitive for the determination of triterpenoids in Ilicis Hainanensis Folium, and may be used for the quality improvement of this herb.

AIM:To investigate the effects of baicalein on pulmonary arterial hypertension(PAH)induced by monocrotaline(MCT)in rats,and its molecular mechanism was further explored.METHODS: Male SD rats(n=28) were randomly divided into 4 groups: control group, MCT group, MCT+baicalein 50 mg/kg group and MCT +baicalein 100 mg/kg group.The PAH model was established by subcutaneous injection of MCT.After 2 weeks of modeling,the rats in baicalein treatment groups were gavaged baicalein 50 and 100 mg· kg -1· d-1for 14 d,the rats in control group were administered with saline.After 4 weeks of modeling,right ventricular systolic pressure(RVSP),right ventricular hypertro-phy index(RVHI)and right ventricular mass index(RVMI)were detected.Masson staining was used to detect the degree of lung fibrosis.The pathomorphological changes of the pulmonary vessels were observed by HE staining.Western blot was used to detect the expression of α-smooth muscle actin(α-SMA)in the lung tissue and the phosphorylation p 38,ERK and JNK in the artery.RESULTS:Compared with the control group,RVSP, RVHI and RVMI increased significantly in the MCT group(P<0.01).Pulmonary fibrosis and the thickening of pulmonary artery wall were observed.α-SMA was up-regulated and p38,ERK and JNK was activated significantly(P<0.01).Compared with the MCT group,baicalein(50 and 100 mg/kg)significantly decreased the RVSP,RVHI and RVMI(P<0.01).Lung fibrosis was reduced and the vas-cular wall thickening was decreased in baicalein-treated groups.Baicalein(50 and 100 mg/kg)inhibited the phosphoryla-tion of p38,ERK and JNK compared with the MCT group(P<0.01).CONCLUSION:Baicalein ameliorates MCT-in-duced PAH by the inhibition of pulmonary artery wall thickening at least partially via MAPK signaling pathway.