Effect of Gupi Xiaoji Decoction on mitochondrial structure and function of human hepatoma cell line HepG2 / 中国药理学通报

Aim To observe the effect of Gupi Xiaoji Decoction (GPXJY) on the structure and function of mitochondria of human hepatoma cell HepG2 cells and explore its possible mechanism. Methods CCK8 was used to detect cell proliferation, Mito-Tracker Green fluorescence staining was used to observe the mitochondrial structure, flow cytometry was used to detect the membrane potential, Elisa was used to detect the ATP content, fluoroscopic electron microscopy was used to observe the microstructure changes, and high-content screening(HCS) was used to detect the related proteins. Results Fluorescence staining showed that GPXJY damaged the mitochondria of HepG2 cells and decreased the content of ATP. The results of flow cytometry showed that GPXJY could reduce the mitochondrial membrane potential of HepG2 cells. The results of electron microscope showed that GPXJY made the mitochondria of cancer cells swell and so on. HCS found that GPXJY significantly reduced the average fluorescence intensity of Bcl-2 in HepG2 cells, and significantly increased the average fluorescence intensity of apoptosis promoting proteins Bax, cytochrome-c, caspase-3 and cleaved-caspase-3, which was statistically significant. Conclusion GPXJY can regulate the structure and function of mitochondria in HepG2 cells.

Effect of Gupi Xiaoji Decoction on Pyroptosis of HepG2.2.15 Cells Based on Network Pharmacology and Molecular Docking / 中国实验方剂学杂志

ObjectiveTo screen the active antitumor components of Gupi Xiaoji decoction by network pharmacology and molecular docking based on the pyroptosis mediated by cysteinyl aspartate-specific protease 1 （Caspase-1） and explore its molecular mechanism in intervening in the pyroptosis of HepG2.2.15 cells through in vitro experiments. MethodThe compounds and targets of Gupi Xiaoji decoction were screened out by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） to obtain the corresponding gene symbols. The targets of Caspase-1 were collected from GeneCards，online mendelian inheritance in man（OMIM），PharmGKB，and TTD，and the compound-gene target regulatory network was constructed by Cytoscape. The protein-protein interaction（PPI） network was established and analyzed by STRING. The mechanism of the effective components of Gupi Xiaoji decoction on Caspase-1 was predicted by gene ontology（GO） functional enrichment and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analyses. The molecular docking was verified with AutoDock Vina. The plasma medicated with Gupi Xiaoji Decoction was prepared and HepG2.2.15 cells were cultured in vitro. HepG2.2.15 cells were divided into a blank plasma group，a VX-765 group，a VX-765+medicated plasma group， and a medicated plasma group. After 48 hours of intervention with 15% medicated plasma， the expression and distribution of gasdermin D-N （GSDMD-N） on the surface of the cell membrane were detected by immunofluorescence staining. The release of lactic dehydrogenase （LDH）， interleukin（IL）-1β，and IL-18 in the cell supernatant was measured by enzyme-linked immunosorbent assay（ELISA） kits. The expression of Caspase-1 and GSDMD-N was measured by Western blot. ResultThe mitogen-activated protein kinase 14 （MAPK14），MAPK1，protein kinase B1 （Akt1）， MAPK8， V-Jun sarcoma virus oncogene homolog （JUN）， and TP53 screened by network pharmacology were the main targets. The compounds 7-hydroxy-5，8-dimethoxy-2-phenyl-chromone，wogonin，rhamnazin，moslosooflavone，isorhamnetin，7-O-methylisomucronulatol，formononetin，calycosin，luteolin，quercetin，kaempferol，β-sitosterol，and baicalein screened by network pharmacology were the main active components of Gupi Xiaoji decoction. Go enrichment analysis showed that multiple biological processes were involved， including responses to oxidative stress and metal ions，ubiquitin-like protein ligase binding，and phosphatase binding. KEGG pathway enrichment analysis showed MAPK pathway，nuclear factor（NF）-κB pathway，p53 pathway， and hypoxia-inducible factor-1（HIF-1） pathway were involved. Molecular docking showed that the targets had good binding with the components. In vitro experiments displayed that compared with the blank plasma group，the VX-765 group showed weakened GSDMD-N fluorescence signal，reduced release of LDH，IL-1β，and IL-18，and declining expression of Caspase-1 and GSDMD-N（P<0.01）， and the medicated plasma group showed increased GSDMD-N fluorescence signal， increased release of LDH，IL-1β，and IL-18，and up-regulated expression of Caspase-1 and GSDMD-N（P<0.01）. ConclusionGupi Xiaoji Decoction can induce the pyroptosis of HepG2.2.15 cells by regulating Caspase-1 through multiple targets and multiple pathways.

Data Mining and Systems Pharmacology to Elucidate Effectiveness and Mechanisms of Chinese Medicine in Treating Primary Liver Cancer / 中国结合医学杂志

OBJECTIVE@#To identify specific Chinese medicines (CM) that may benefit patients with primary liver cancer (PLC), and to explore the mechanism of action of these medicines.@*METHODS@#In this retrospective, singlecenter study, prescription information from PLC patients was used in combination with Traditional Chinese Medicine Inheritance Supports System to identify the specific core drugs. A system pharmacology approach was employed to explore the mechanism of action of these medicines.@*RESULTS@#Taking CM more than 6 months was significantly associated with improved survival outcomes. In total, 77 putative targets and 116 bioactive ingredients of the core drugs were identified and included in the analysis (P<0.05). A total of 1,036 gene ontology terms were found to be enriched in PLC. A total of 75 pathways identified from Kyoto Encyclopedia of Genes and Genomes were also enriched in this disease, including fluid shear stress, interleukin-17 signaling, signaling between advanced glycan end products and their receptors, cellular senescence, tumor necrosis factor signaling, p53 signaling, cell cycle signaling, steroid hormone biosynthesis, T-helper 17 cell differentiation, and metabolism of xenobiotics by cytochrome. Docking studies suggested that the ingredients in the core drugs exert therapeutic effects in PLC by modulating c-Jun and interleukin-6.@*CONCLUSIONS@#Receiving CM for 6 months or more improves survival for the patients with PLC. The core drugs that really benefit for PLC patients likely regulates the tumor microenvironment and tumor itself.

Preliminary clinical application of complete digital workflow of design and manufacturing occlusal splint for sleep bruxism / 北京大学学报(医学版)

OBJECTIVE@#To establish a complete workflow of digital design and manufacturing occlusal splint for sleep bruxism, which can be preliminarily applied in clinical use, thus observe the clinical efficacy.@*METHODS@#Twenty-four patients with sleep bruxism were recruited in the study and randomly divided into two groups by using random number tables. Digital-occlusal-splint (experimental group) treatment plan and traditional-occlusal-splint (control-group) treatment plan were carried out for each group, respectively. For experimental group, digital models of patients' both dental arches and the occlusion relationship after elevation were captured using an intraoral scanner. The occlusal splint was carried out by computer aided design/computer aided manufacturing (CAD/CAM), including splint designing and milling. For control group, the traditional soft occlusal splint was fabricated by vacuum laminator. The two kinds of occlusal splints were tried in the patients from each group, and the occlusal contacts were tested respectively by T-scan analysis system, which recorded the changes of occlusal indicators in the two groups. The retention, appearance and occlusal comfort degree were evaluated by the two groups of patients. Mann-Whitney test was performed with IBM SPSS 20.0 software, and bilateral test was performed. P<0.05 was considered to be statistically significant.@*RESULTS@#The complete workflow of digital design and manufacturing occlusal splint was successfully established. During the clinical use, there was no statistical difference in the retention evaluation of two kinds of occlusal splints between the two groups of patients (Z=-0.538, P=0.590). The appearance score (Z=2.038, P=0.042) and the occlusal comfort score (Z=-2.579, P=0.010) of the experimental group were higher than those of the control group, with statistically significant differences. The T-scan analysis results showed that only the second molar on both sides of the traditional occlusal splint had occlusal contact in intercupsal position, while the digital occlusal splint had stable and bilaterally balanced contact between the maxillary and mandibular teeth. Furthermore, the occlusal force was uniformly distributed in the experimental group.@*CONCLUSION@#The complete workflow of digital occlusal splint improves the occlusal design, greatly simplifies and optimizes the traditional process of making occlusal splint. This new method is resource-saving and environmental-friendly, and it is able to serve patients more conveniently and efficiently.

Evaluation of the fabrication deviation of a kind of milling digital implant surgical guides / 北京大学学报(医学版)

OBJECTIVE@#To evaluate the deviation of digital implant surgical guides during fabrication process in the Organical Dental Implant (ODI) system.@*METHODS@#This study included two parts. The first part was the in vitro study. A resin block with a diagnostic template was used for the planning. After cone beam computed tomography (CBCT) scanning, a surgical guide with eight implants was virtually designed using the ODI system. The guide was milled by a 5-axial numerical controlled milling machine, and an optical scanning was taken to digitalize the guide to a standard tessellation language (STL) form. The STL data were then imported into an ODI software and registered with the original design. The deviation of the sleeves between the design and the STL was measured in the ODI software and set as the golden standard. Then the ODI examination table was used to measure the deviation of the guide during fabrication. Examiners A and B measured 10 times separately. The reliability and the validity of the examination table was calculated. The second part was the in vivo study: The deviation during fabrication of 12 guides designed and fabricated by the ODI system were measured using the examination table.@*RESULTS@#The standard deviation of the deviation measured using the examination table by examiners A and B were all below 0.40 mm (for the shell reference points) and 0.71 degree (for the angles). No significant difference was found between the two examiners for any implant sites. The result of the examination table was larger than that of the software for the shell reference point (t-test, P<0.05), but no significant difference was found for the angle deviation (t-test, P>0.05). The 45 implants positions in the 12 guides for the in vivo study were examined using the examination table. The deviations at the shell reference points were (1.06±0.29) mm (0.42-1.75 mm), and at the implant tip were (1.12±0.48) mm (0.41-2.44 mm). The angle deviations were (1.42±0.70) degree (0.29-2.96 degree).@*CONCLUSION@#Deviation is unavoidable during the fabrication process of the guides. The examination table of the ODI system is a reliable and valid tool to measure the deviation during fabrication of the ODI guides. More studies should be designed to research the relationship between the fabrication deviation and the implant insertion deviation.

Progress of traditional medicines for treatment of ischemic stroke / 中国药理学与毒理学杂志

Ischemic stroke has the characteristics of high morbidity and high mortality, which seriously endanger human health.According to the statistics, ischemic stroke in China accounts for about 80% of stroke cases, and its mortality rate is as high as 50%, but most of the ischemic stroke patients who survive have sequelae, such as hemiplegia, aphasia and so on.In recent years, people have continuously studied the pathological cascade of ischemic stroke, and also achieved some good results in animal model experiments, however, the clinical results are not satisfactory. Nowadays, the better clinically effective therapeutic drug is tissue plasminogen activator, but due to the limited time window,only a small number of patients can apply this drug in time to achieve the effect.Therefore,it is essential to study drugs that are definitive and can benefit a large number of patients with ischemic stroke.The development of traditional medicines is not as fast as the development of new drugs. For ischemic stroke,we can also turn from the study of the pathogenesis of ischemic stroke to the study of traditional drugs for the endogenous neuroprotection of ischemic stroke, and look for effective targets for neuroprotection based on traditional medicine.The multi-targets and multi-effects for the acute and convalescent phases of ischemic stroke is a direction explored by researchers, which also provides some evidence for more effective drugs for the treatment of ischemic stroke.

Effects of Yiqi Huayu Jiedu Prescription on MHCC97-H Migration Ability and Expressions of Chemokines CXCL12, CXCR4 and CXCR7 / 中国中医药信息杂志

Objective To study the effects of Yiqi Huayu Jiedu Prescription on the migration ability of MHCC97-H and the expressions of CXCL12, CXCR4 and CXCR7; To discuss its relevant mechanism of action. Methods Setting Sorafenib as a positive control, CCK-8 method was used for determining the effects of Yiqi Huayu Jiedu Prescription on the cell proliferation of MHCC97-H and the optimum concentration. Scratch assay was used to observe the migration ability of MHCC97-H. The protein expressions of CXCL12, CXCR4 and CXCR7 were detected by Western blot after 24 h of medicine intervention. Results Yiqi Huayu Jiedu Prescription and Sorafenib can inhibit the cell proliferation of MHCC97-H , and the inhibitory concentration was 0.095 g/mL and 10 μmol/mL at 24 hours. Yiqi Huayu Jiedu Prescription can inhibit migration ability of MHCC97-H. The protein expressions of CXCL12, CXCR4 and CXCR7 in hepatocellular carcinoma cells decreased after the action of Yiqi Huayu Jiedu Prescription. Conclusion Yiqi Huayu Jiedu Prescription can inhibit MHCC97-H cell proliferation and migration, which may be realized by down-regulating chemokine axis of CXCL12/CXCR4/CXCR7.

A Risk Prediction Model for In-hospital Mortality in Patients with Suspected Myocarditis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Myocarditis is an inflammatory disease of the myocardium that may lead to cardiac death in some patients. However, little is known about the predictors of in-hospital mortality in patients with suspected myocarditis. Thus, the aim of this study was to identify the independent risk factors for in-hospital mortality in patients with suspected myocarditis by establishing a risk prediction model.</p><p><b>METHODS</b>A retrospective study was performed to analyze the clinical medical records of 403 consecutive patients with suspected myocarditis who were admitted to Ningbo First Hospital between January 2003 and December 2013. A total of 238 males (59%) and 165 females (41%) were enrolled in this study. We divided the above patients into two subgroups (survival and nonsurvival), according to their clinical in-hospital outcomes. To maximize the effectiveness of the prediction model, we first identified the potential risk factors for in-hospital mortality among patients with suspected myocarditis, based on data pertaining to previously established risk factors and basic patient characteristics. We subsequently established a regression model for predicting in-hospital mortality using univariate and multivariate logistic regression analyses. Finally, we identified the independent risk factors for in-hospital mortality using our risk prediction model.</p><p><b>RESULTS</b>The following prediction model for in-hospital mortality in patients with suspected myocarditis, including creatinine clearance rate (Ccr), age, ventricular tachycardia (VT), New York Heart Association (NYHA) classification, gender and cardiac troponin T (cTnT), was established in the study: P = ea/(1 + ea) (where e is the exponential function, P is the probability of in-hospital death, and a = -7.34 + 2.99 × [Ccr <60 ml/min = 1, Ccr ≥60 ml/min = 0] + 2.01 × [age ≥50 years = 1, age <50 years = 0] + 1.93 × [VT = 1, no VT = 0] + 1.39 × [NYHA ≥3 = 1, NYHA <3 = 0] + 1.25 × [male = 1, female = 0] + 1.13 × [cTnT ≥50 μg/L = 1, cTnT <50 μg/L = 0]). The area under the receiver operating characteristic curve was 0.96 (standard error = 0.015, 95% confidence interval [CI]: 0.93-0.99). The model demonstrated that a Ccr <60 ml/min (odds ratio [OR] = 19.94, 95% CI: 5.66-70.26), an age ≥50 years (OR = 7.43, 95% CI: 2.18-25.34), VT (OR = 6.89, 95% CI: 1.86-25.44), a NYHA classification ≥3 (OR = 4.03, 95% CI: 1.13-14.32), male gender (OR = 3.48, 95% CI: 0.99-12.20), and a cTnT level ≥50 μg/L (OR = 3.10, 95% CI: 0.91-10.62) were the independent risk factors for in-hospital mortality.</p><p><b>CONCLUSIONS</b>A Ccr <60 ml/min, an age ≥50 years, VT, an NYHA classification ≥3, male gender, and a cTnT level ≥50 μg/L were the independent risk factors resulting from the prediction model for in-hospital mortality in patients with suspected myocarditis. In addition, sufficient life support during the early stage of the disease might improve the prognoses of patients with suspected myocarditis with multiple risk factors for in-hospital mortality.</p>

Efficacy Comparison of Different Salvage Treatment Regimens for Patients with Refractory/Relapsed Acute Myeloid Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To compare the efficacy and safety of 3 different regimens, namely MAC, FLAG and CAG, as the re-induction chemotherapy for acute myeloid leukemia(AML) patients with primary induction failure and relapse.</p><p><b>METHODS</b>The clinical data of 156 AML patients with primary induction failure and relapse, except patients with acute promyelocytic leukemia(APL), treated with any of the above 3 regimens in our center from January 2008 to April 2016 were analyzed retrospectively. According to the treatment regimens, 156 patients were divided into MAC group (n=60), FLAG group (n=45) and CAG group (n=51). The complete remission(CR), partial remissison(PR), overall survival(OS), disease-free survival(DFS) and adverse events during the treatment were analyzed, so as to compare and evaluate the efficacy and safety of the 3 different regimens.</p><p><b>RESULTS</b>After 1 course of re-induction chemotherapy, CR in MAC group was significantly higher than that in FLAG and CAG group (55.4% vs 34.1% vs 34.0%)(P<0.05). The OS was not statistically significantly different among 3 groups (P>0.05) with a median OS of 11 months, 5.46 months and 10.2 months, respectively. The myelosuppression was the main adverse event with no significant difference among the groups(P>0.05). More patients treated with MAC regimen underwent febrile neutropenia (93.3% vs 86.7% vs 64.7%)(P<0.001). However, the incidence of fatal infections was not signicantly different among 3 groups(5% vs 8.9% vs 5.9%)(P>0.05).</p><p><b>CONCLUSION</b>Compared with FLAG and CAG regimen, the MAC regimen can enable more AML patients with primary induction failure and refractory to achieve CR without increasing severe adverse events,therefore,this regimen may provide a opportunity for patients to recieve hematopoietic stem cell transplantation.</p>

Preliminary Clinical Efficiency of Autologous Peripheral Blood Mononuclear Cells for Treating Critical Limb Ischemia of Thromboangiitis Obliterans / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To evaluate the long-term clinical effect of autologous peripheral blood mononuclear cells (PB-MNC) on critical limb ischemia (CLI) in patients with thromboangiitis obliterans (TAO) patients.</p><p><b>METHODS</b>The clinical data of 22 patients with CLI caused by TAO from July 2004 to May 2013 were analyzed retrospectively, 22 patients were divided into 2 groups; out of them 12 cases in one group were treated with granulocyte colony-stimulating factor (G-CSF)-mobilized autologous peripheral blood mononuclear cells (auto-PBMNC group), 10 cases in another group received conservative treatment (CT group). The log-rank test was used to compare the long-term outcomes in auto-PBMNC group and CT group.</p><p><b>RESULTS</b>The wound healing rate (P=0.016) and CLI-free rate (P=0.013) were significantly higher in PB-MNC group compared with that in CT group. No difference was found in amputation rates between the 2 groups (major amputation: P=0.361, minor and major amputation: P=0.867). No patients died or no serious adverse events occurred during the follow-up period.</p><p><b>CONCLUSION</b>The auto-PBMNC therapy can significantly promote the wound healing, and protect against CLI in TAO patients, but the risk of amputation is not low in comparison with conservative treatment.</p>

Pharmacokinetics of amphotericin B in the cerebrospinal fluid during continuous intrathecal administration for treatment of cryptococcal neoformans meningitis / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the pharmacokinetics of amphotericin B (AMB) in the cerebrospinal fluid (CSF) during continuous intrathecal administration of AMB for treatment of cryptococcal neoformans meningitis (CNM).</p><p><b>METHODS</b>The concentration of AMB in the CSF was measured using reversed phase high performance liquid chromatography (RP-HPLC) in 3 patients receiving continuous intrathecal infusion of AMB for CNM.</p><p><b>RESULTS</b>AMB concentrations in the CSF of the 3 patients exceeded the minimal inhibitory concentration (MIC) of AMB against Cryptococcus neoformans. The concentration-time curve showed that AMB concentration in the CSF underwent obvious variations on the first day of intrathecal infusion and after additional AMB doses, but maintained a stable level (0.61-1.21 µg/ml) on the next day.</p><p><b>CONCLUSION</b>[corrected] Continuous intrathecal administration of AMB can enhance the drug concentration in the CSF and maintain a stable and effective drug level for treatment of CNM.</p>

Pharmacokinetics, tissue distribution and magnetic resonance's response characterstics of folic acid-O-carboxymethyl chitosan ultrasmall superparamagnetic iron oxide nanoparticles in mice and rats / 药学学报

Folic acid-O-carboxymethyl chitosan ultrasmall superparamagnetic iron oxide nanoparticles (FA-OCMCS-USPIO-NPs) are a novel molecular targeting MR contrast agent. This paper reperts the pharmacokinetics and magnetic resonance response characteristics of FA-OCMCS-USPIO-NPs in normal rats and mice, and discussed its distributing regularity in animals, providing basis for tumor targeting imaging. O-phenanthroline method was used to determine iron content in rats' plasma and mice's organs following high and low doses of nanoparticles injected through tail vein, and the blood concentration-time curve was drawn, the calculated t1/2 of two groups were greater than 7 h. The results of tissue distribution showed that only a small part of nanoparticles were swallowed by the liver and spleen, while none in the heart, lung and kidney. At the same times, the phagocytosis of nanoparticles did not change with the dose. The results of MRI showed that renal excretion occurred 4 hours after injection, and signal to noise ratio (SNR) of liver and kidney returned to normal levels 24 hours after injection. There were no nanoparticles in the lungs. So a part of nanoparticles escaped from phagocytosis of liver and spleen, and it owned lower toxicity and longer half-life. indicated its use for tumor-targeting imaging. All of these indicated its use for tumor-targeting imaging.