Evolution and development of potent monobactam sulfonate candidate IMBZ18g as a dual inhibitor against MDR Gram-negative bacteria producing ESBLs

A series of new monobactam sulfonates is continuously synthesized and evaluated for their antimicrobial efficacies against Gram-negative bacteria. Compound 33a (IMBZ18G) is highly effective in vitro and in vivo against clinically intractable multi-drug-resistant (MDR) Gram-negative strains, with a highly druglike nature. The checkerboard assay reveals its significant synergistic effect with β-lactamase inhibitor avibactam, and the MIC values against MDR enterobacteria were reduced up to 4-512 folds. X-ray co-crystal and chemoproteomic assays indicate that the anti-MDR bacteria effect of 33a results from the dual inhibition of the common PBP3 and some class A and C β-lactamases. Accordingly, preclinical studies of 33a alone and 33a‒avibactam combination as potential innovative candidates are actively going on, in the treatment of β-lactamase-producing MDR Gram-negative bacterial infections.

Discovery and identification of EIF2AK2 as a direct key target of berberine for anti-inflammatory effects

Using chemoproteomic techniques, we first identified EIF2AK2, eEF1A1, PRDX3 and VPS4B as direct targets of berberine (BBR) for its synergistically anti-inflammatory effects. Of them, BBR has the strongest affinity with EIF2AK2 via two ionic bonds, and regulates several key inflammatory pathways through EIF2AK2, indicating the dominant role of EIF2AK2. Also, BBR could subtly inhibit the dimerization of EIF2AK2, rather than its enzyme activity, to selectively modulate its downstream pathways including JNK, NF-κB, AKT and NLRP3, with an advantage of good safety profile. In EIF2AK2 gene knockdown mice, the inhibitory IL-1β, IL-6, IL-18 and TNF-α secretion of BBR was obviously attenuated, confirming an EIF2AK2-dependent anti-inflammatory efficacy. The results highlight the BBR's network mechanism on anti-inflammatory effects in which EIF2AK2 is a key target, and inhibition of EIF2AK2 dimerization has a potential to be a therapeutic strategy against inflammation-related disorders.

Synthesis and biological evaluation of novel tricyclic matrinic derivatives as potential anti-filovirus agents

Twenty-six novel tricyclic sophoridinic and matrinic derivatives containing a common chlorinated benzene fragment were designed, synthesized and evaluated for their anti-ebolavirus (EBOV) activities. Structure-activity relationship analysis indicated: (i) 12-dichlorobenzyl motif was beneficial for the activity; (ii) the chiral configuration at C5 atom might not affect the activity much. Among the target compounds, compound exhibited the most potent potency against EBOV with an IC value of 5.29 μmol/L and an SI value of over 37.8. Further anti-EBOV assay of identified its high effectiveness, and anti-MARV assay of suggested its inspiring broad-spectrum anti-filovirus activity. The results provided powerful information on further strategic optimization and development of this kind of compounds against filoviruses.

AEG-1 promotes metastasis of breast cancer MCF-7 cells / 实用肿瘤学杂志

Objective The objective of this study was to observe the effect of AEG-1 gene on the metastasis in breast cancer MCF-7 cells.Methods AEG-1 siRNAs were transfected into MCF-7 cells to silence AEG-1 expression,and negative siRNA was used as a control.Transwell chamber was used to detect the ability of cell migration and invasion of MCF-7 cells.CCK8 assay was used to detect the cell proliferation of MCF-7 cells.At the same time,the effect of VEGF on the angiogenesis was investigated by detecting the changes of lumen formation in HUVEC cells.Results The migration,invasive and proliferative abilities were significantly inhibited in MCF-7 cells transfected with AEG-1 siRNA.Knockdown AEG-1 was significantly decreased the level of VEGF in the supernatant of MCF-7 cells.Knockdown AEG-1 was also significantly inhibited the angiogenesis activity in HUVEC cells.Conclusion Knockdown AEG-1 can significantly inhibit the migration of MCF-7 cells,including cell migration,invasion,proliferation and angiogenesis.These results suggest that AEG-1 plays an important role in the metastasis process of breast cancer and opens up new ideas for future treatment breast cancer.

Synthesis and biological evaluation of 12-N-p-chlorobenzyl sophoridinol derivatives as a novel family of anticancer agents

Taking 12-N-p-chlorobenzyl sophoridinol 2 as a lead, a series of novel sophoridinic derivatives with various 3'-substituents at the 11-side chain were synthesized and evaluated for their anticancer activity from sophoridine (1), a natural antitumor medicine. Among them, the sophoridinic ketones 5a-b, alkenes 7a-b and sophoridinic amines 14a-b displayed reasonable antiproliferative activity with IC50 values ranging from 3.8 to 5.4 μmol/L. Especially, compounds 5a and 7b exhibited an equipotency in both adriamycin (AMD)-susceptible and resistant MCF-7 breast carcinoma cells, indicating a different mechanism from AMD. The primary mechanism of action of 5a was to arrest the cell cycle at the G0/G1 phase, consistent with that of parent compound 1. Thus, we consider 12-chlorobenzyl sophoridinic derivatives with a tricyclic scaffold to be a new class of promising antitumor agents with an advantage of inhibiting drug-resistant cancer cells.

Synthesis and biological evaluation of sophocarpinic acid derivatives as anti-HCV agents

Chronic hepatitis C virus (HCV) infection has become a major public health burden worldwide. Twenty-two sophocarpinic acid or matrine derivatives were synthesized and their anti-HCV activities were evaluated in vitro. The structure-activity analysis revealed that (i) sophocarpinic acids with a D-seco 3-ring structure scaffold were more favorable than matrines with a 4-ring scaffold; (ii) the introduction of an electron-withdrawing group on the phenyl ring in 12-N-benzenesulfonyl Δ (βγ) sophocarpinic acids was beneficial for the antiviral activity against HCV. Among them, compounds 9h and 9j exhibited the most potent inhibitory activities on HCV replication with selectivity indies of 70.3 and 30.9, respectively. Therefore, both were selected as antiviral candidates for further investigation.

Effect of Broussonetia papyrifera ( L. ) Vent aqueous extract on the ability of space learning and memory in the rats complex model of Alzheimer's disease / 中华行为医学与脑科学杂志

Objective To observe the effect of aqueous extract of Broussonetia papyrifera ( L. ) Vent on the ability of space learning and memory in the rats with Alzheimer' s disease (AD) induced by Aβ 25-35 and Dgalactose and to explore the mechanisms underlying those improvements. Methods The animal model of AD was established by Aβ 25-35 stereotactic injection into the hippocampus of rats in 5 minutes,while long-term intraperitoneal injection with D-gal. After the injection of Aβ25-35,rats were treated with aqueous extract of Broussonetia papyrifera ( L. ) Vent for the next 30 days. Morris water maze with computer system and the spatial exploration experiments were used to assess the behavior performances of the rats. Immunohistochemical technique was used to detect the expression of BiP, PERK and CHOP. Results The ability of space learning and memory of rats complex model of Alzheimer's disease induced by Aβ25-35 and D-gal was damaged,while escape latency was (20.90± 9.16 ) s,and the proportion of original platform quadrant was ( 11.05 ± 4.43 ) %. The expression level of Bip was reduced ,while the mean gray was ( 139.71 ± 3.47 ). The expression level of PERK and CHOP was increased,while the mean gray were (97.96 ± 5.97 ), ( 110.93±4.91 )separately. The escape latency of rats in the aqueous extract of Broussonetia papyrifera ( L. ) Vent treated groups was ( 5.41 ± 3.47 ) s and shorter than the model group,while the proportion of original platform quadrant was (48.28 ± 7.03 )% and higher than the model group.The expression level of Bip in the treated group was higher than the model group, while the mean gray were ( 121.17 ±4.76). The expression level of PERK and CHOP in the treated group was lower than the model group,while the mean gray were ( 122.11 ± 4.73 ), ( 123.34 ± 7.73 ) separately. Significant differences were observed between model group and aqueous extract of Broussonetia papyrifera ( L. ) Vent treated groups (P＜ 0. 05 ～ 0. 01 ).Conclusion Aqueous extract of Broussonetia papyrifera ( L. ) Vent can improve learning and memory disorders of the model rats induced by Aβ25-35 and D-galactose. ER (endoplasmic reticulum) stress and correlated apoptosis pathway might be involved in the underlying mechanisms.

Synthesis and structure-activity relationship of 13-hexylberberine analogues as CD36 antagonists / 药学学报

Scavenger receptor CD36 could bind and endocytose oxLDL into macrophages which were then differentiated into foam cells that constitute the atherosclerotic lesion core, and was considered to be a potential target to treat atherosclerosis. In the establishment of the compound library of berberine (BBR, 1) analogues, we discovered that 13-hexylberberine (2) showed an antagonistic activity against CD36. Taking 2 as the lead compound, 21 derivatives were synthesized and their antagonistic activities were evaluated via an ELISA-like high-throughput screening (HTS) model. The primary structure-activity relationships were studied. It was indicated that the introduction of suitable groups at the 2- and 3-position of the aromatic ring A or at the 9-position of the aromatic ring D could enhance the activity. Among the 21 studied compounds, 7g bearing a benzyloxyl group at the 9-position provided a highest CD36 antagonistic activity with the IC50 value of 7.7 micromol L(-1). Besides, its antagonistic activity was further verified with Sf9 insect cell HTS model. So berberine analogues are a new family of CD36 receptor antagonists and worthy to be studied further.

Study on the Feasibility for Judging the Lifting-thrusting Manipulation of Acupuncture by Using "Quality Control Figure" Measurement / 针刺研究

0.05).But a positive correlation was found between them(r =0.6943,P

In vitro isolation of intestinal side population cells from newborn mice / 中国组织工程研究

BACKGROUND: Current methods of stem cell separation are mainly based on their cell markers.A method for stem cells separation which is not based on cell markers developed in recent years,that is fluorescence activated cell sorting method,has been applied for stem cells and mature cells separation.OBJECTIVE: To isolate side population cells from newborn mice small intestinal mucosa,and to investigate the feasibility of constructing the murine intestinal stem cell population by fluorescence activated cell sorting.METHODS: Small intestine mucosa organoids of mice were isolated and dissociated into single cells.The side population cells were stained with Hoechst 33342 and propidium iodide,then sorted using fluorescence activated cell sorting.Total RNA and protein were purified from sorted fractions to detect Musashi-1 expressions by RT-PCR and Western-blotting.RESULTS AND CONCLUSION: Single cell suspension from mouse small intestine mucosa contained a viable population of cells,which showed the side population phenotype and were sensitive to verapamil.These cells were enriched for Musashi-1 mRNA and MSI-1 protein expression.Results demonstrated that the side population fraction separated from mice intestinal mucosa is enriched for intestinal stem cells,the murine intestinal stem cell population can be successfully constructed with fluorescence activated cell sorting.

Function and Significance of Very Low Density Lipoprotein Receptor Subtype Ⅱ / 华中科技大学学报（医学）（英德文版）

Summary: To explore the functions of very low density lipoprotein receptor (VLDL-R) subtype II in lipoprotein metabolism and foam cells formation, the recombinant plasmid with the two subtypes cDNA was constructed respectively, the ldl-A7 cell lines were transfected and two cell lines expressing VLDL-R were obtained: one stably expressing the VLDLR with the O-linked sugar region (type I VLDLR) and the other without the O-linked sugar region (type II VLDLR). In the study on binding of VLDLR to their nuclein labeled natural ligands (VLDL and β-VLDL), it was found that surface binding of 125I-VLDL or 125I-β-VLDL of ldl-A7 cells transfected with type I VLDLR recombinant (ldl-A7-VRI) was more higher than that of ldl-A7 cells transfected with type II VLDLR recombinant (ldl-A7-VRII). After being incubated with VLDL for different time, the contents of triglyceride and total cholesterol in cells were mensurated, and the formation of foam cells and accumulation of lipid in cells was observed by oil-red O staining. The results showed that the contents of triglyceride and total cholesterol in ldl-A7-VR I were much higher than those in ldl-A7-VR II, and ldl-A7-VR I could transform into foam cells notably. It was suggested that type I VLDLR binds with relative higher affinity to VLDL and β-VLDL, and internalizes much more lipoprotein into cells. As a result, we can conclude that type I VLDLR plays a more important role in lipoprotein metabolism and foam cells formation than type II VLDLR.

Effects of Oxidized Low Density Lipoprotein on the Expression and Function of ABCA1 in Macrophages / 华中科技大学学报（医学）（英德文版）

In the present study, we examined the regulation of the expression and function of AB CA1 by modified LDL (ox-LDL) in vitro. After incubation with apoA-Ⅰ for 24 h, RAW264.7 cells effluxed 37.65 % cholesterol loaded by acetyl LDL (ac-LDL), and 9.78 % cholesterol in ox-LDL group. The level of ABCA1 Mrna increased about three times either when cells were incubated with 100 μg/Ml ac-LDL or with 100μg/Ml ox-LDL. However, the level of ABCA1 protein rose by 1.57 times in ac-LDL group and 1.26 times in ox-LDL group. These results demonstrated that ox-LDL had different effect on the expression and function of ABCA1, ox-LDL might decrease the cholesterol efflux mediated by ABCA1 through other unknown mechanisms.

Role of VLDL receptor in the process of foam cell formation / 华中科技大学学报（医学）（英德文版）

The role of very low density lipoprotein receptor (LVLDR) in the process of foam cell formation was investigated. After the primary cultured mouse peritoneal macrophages were incubated with VLDL, beta-VLDL or low density lipoprotein (LDL), respectively for 24 h and 48 h, foam cells formation was identified by oil red O staining and cellular contents of triglyceride (TG) and total cholesterol (TC) were determined. The mRNA levels of LDLR, LDLR related protein (ILRP) and VLDLR were detected by semi-quantitative RT-PCR. The results demonstrated that VLDL, beta-VLDL and LDL could increase the contents of TG and TC in macrophages. Cells treated with VLDL or beta-VLDL showed markedly increased expression of VLDLR and decreased expression of LDLR, whereas LRP was up-regulated slightly. For identifying the effect of VLDL receptor on cellular lipid accumulation, ldl-A7-VR cells, which expresses VLDLR and trace amount of LRP without functional LDLR, was used to incubate with lipoproteins for further examination. The results elucidated that the uptake of triglyceride-rich lipoprotein mediated by VLDLR plays an important role in accumulation of lipid and the formation of foam cells.

Interference effects of tetramethylpyrazine on the proliferation of vascular smooth muscle cells(VSMCs) induced by angiotensin Ⅱ / 解放军医学杂志

Objective To study the interference effects of tetramethylpyrazine (TMP) on calcinuerin (CaN), c-fos and the nuclear antigen of proliferating cells in the proliferation of vascular smooth muscle cells (VSMCs) treated by angiotensinⅡ(Ang Ⅱ). Methods A cell proliferating model of VSMCs induced by Ang Ⅱ was established; the effects of TMP on CaN was detected by enzyme reaction phosphorus measurement; the effects of TMP on c-fos gene and PCNA expression were observed by immunocytochemical staining and image analysis technique (A value). Results The rats’ aortic smooth muscle cells were cultured in vitro successfully. CaN activities, cell proliferation activity and the expression levels of c-fos and PCNA increased significantly in VSMCs proliferation induced by Ang Ⅱ (P

Effect of cyclosporin A on neuropeptide Y-induced proliferation of cultured vascular smooth muscle cells / 中国病理生理杂志

AIM: To study the role of calcineurin (CaN)-dependent signaling pathway in proliferation of vascular smooth muscle cells (VSMCs) by observing the effect of cyclosporin A (CsA) on proliferation of neuropeptide Y (NPY)-induced rat VSMCs. METHODS: Upon the model of cultured rat VSMCs, the study consisted of three groups: NPYgroup,CsA+NPY group and control group. CaN activity was determinated by enzyme reaction phosphorus measurement. The methods of biochemistry (MTT) and quantitative immunocytochemistry were applied to investigate the proliferation of VSMC and the expression of proliferation cell nuclear antigen (PCNA) in cultured rat VSMCs. (RESULTS:) (Compared) with the control group, the VSMC's CaN activity, proliferation activity and expression of PCNA (by photo densitometry A_(PCNA)) were obviously increased in NPY group (P