Mechanisms of Panax ginseng in preventing rat pheochromocytoma cells from apoptosis

Aims of the study

Although ginseng root possesses dominant central therapeutic effects and has recently undergone investigations for treating different neuronal diseases, most of its mechanisms are still unknown. Therefore, the neuroprotective mechanisms of ginseng were studied.

Materials and methods

The protection afforded by different methanol extracts of Panax ginseng (PG) was tested in a serum deprivation-induced apoptotic model using neuronal-like pheochromocytoma (PC12) cells. An MTT assay, annexin V-FITC staining, and Western blots were, respectively, applied to identify the viability of cells, the apoptotic form of cell death, and the activity of antiapoptotic signaling.

Results

The known antiapoptotic PI3-K/Akt and MEK/ERK pathways in this system were ruled out due to failure of LY 294002 and PD 98059 to block the protection by PG. A protein kinase A (PKA) inhibitor was found to block the protection by PG and PG-induced CREB phosphorylation, suggesting that the PKA/CREB pathway mediates the protective effect of PG. Downregulation of classical and novel PKCs failed to block the protection by PG, while an atypical PKC inhibitor blocked protection by PG.

Conclusions

PKA and atypical PKC are important for the protection afforded by PG in preventing serum deprivation-induced PC12 cell apoptosis.     

Ligusticum chuanxiong as a potential neuroprotectant for preventing serum deprivation-induced apoptosis in rat pheochromocytoma cells: Functional roles of mitogen-activated protein kinases

Ethnopharmacological relevance

Ligusticum chuanxiong (LC) as a common component in many traditional Chinese medicinal formulas and decoctions has been used to treat different central nervous diseases, suggesting a neuroprotective function.

Aim of the study

To investigate the functional roles of mitogen-activated protein kinases (MAPKs) in mediating the neuroprotection of LC.

Materials and methods

Different extractions of LC were applied with or without MAPK inhibitor to test their protection against serum deprivation-induced apoptosis in rat neuronal-like pheochromocytoma (PC12) cells as revealed by an MTT assay or Hoechst staining. Western blot was used to identify the activations of MAPKs.

Results

The most effective butanol extraction (LC-BuOH) was used in the following experiments. LC-BuOH reversed serum deprivation-induced decreased phosphorylation of extracellular signal-regulated kinase (ERK) and increased phosphorylation of c-Jun NH₂-terminal kinase (JNK) and p38, the family of MAPKs. A PKA inhibitor, blocked the protection of LC-BuOH and partially blocked LC-BuOH-induced alterations in MAPKs, suggesting that the LC-BuOH regulates MAPKs through both PKA-dependent and -independent pathways. Although PD 98059, an inhibitor of MEK which activates ERK, blocked LC-BuOH-induced ERK phosphorylation, it did not block the protection of LC-BuOH.

Conclusions

LC-BuOH mediates protection by suppressing JNK/p38 instead of activating ERK activity.     

丁樱教授治疗IgA肾病经验

丁樱老师为河南中医学院博士研究生导师、教授、主任医师,从事中西医结合儿科教学、临床、科研近40年,治学严谨,造诣深厚,对IgA肾病(IgAN)、紫癜性肾炎、乙肝相关性肾炎、狼疮性肾炎等多种肾脏疾病的治疗具有独到见解.现将导师治疗IgAN的临床经验介绍如下.     

Electro-acupuncture for post-stroke cognitive impairment: A protocol for systematic review and meta-analysis

Background:A considerable number of stroke survivors suffered from cognitive impairment, and more than one third of stroke survivors are affected at 3 and 12 months after the stroke. Although the published systematic reviews suggest that acupuncture can help improve post-stroke cognitive dysfunction, the power of the results is low due to study limitations. Therefore, this review is necessary to analyze the effect of acupuncture on cognitive impairment after stroke and to provide evidence for cognitive impairment in stroke.Methods:This study will be carried out in strict accordance with the Cochrane Handbook for Systematic Reviews of Interventions. According to the pre-established search strategy (PICOS framework), all the literature will be obtained from online databases including Cochrane Central Register of Controlled Trials in the Cochrane Library, Medline (via PubMed), EMBASE (via embase.com), CINAHL (via EBSCOhost), China National Knowledge Infrastructure database, WanFang Database, Chinese Science and Technology Periodical Database, and Sino-Med Database from inception until December 31, 2021 with no language limitations. Two reviewers will screen the records and include quality studies according to inclusion criteria independently. The data needed will be extracted independently by 2 authors according to a table of data extraction. Any inconsistencies in literature screening and data collection will be resolved to reach a consensus via discussion with a third author. Risk of bias for each study will be assessed using risk of bias tool. RevMan5.3 will be used to analyze the data. Heterogeneity will be identified and measured by Chi². Subgroup analyses and sensitivity analysis will be carried out. Grading of Recommendations Assessment, Development and Evaluation will be used to evaluate the evidence for each outcome.Conclusion:The results of this study will provide a theoretical basis for the clinical use of electro-acupuncture to treat cognitive dysfunction after stroke.Unique INPLASY number:INPLASY202210038     

Structure Based Affinity Maturation and Characterizing of SARS-CoV Antibody CR3022 against SARS-CoV-2 by Computational and Experimental Approaches

The COVID-19 epidemic is raging around the world. Neutralizing antibodies are powerful tools for the prevention and treatment of SARS-CoV-2 infection. Antibody CR3022, a SARS-CoV neutralizing antibody, was found to cross-react with SARS-CoV-2, but its affinity was lower than that of its binding with SARS-CoV, which greatly limited the further development of CR3022 against SARS-CoV-2. Therefore, it is necessary to improve its affinity to SARS-CoV-2 in vitro. In this study, the structure-based molecular simulations were utilized to virtually mutate the possible key residues in the complementarity-determining regions (CDRs) of the CR3022 antibody. According to the criteria of mutation energy, the mutation sites that have the potential to impact the antibody affinity were then selected. Then optimized CR3022 mutants with the enhanced affinity were further identified and verified by enzyme-linked immunosorbent assay (ELISA), surface plasma resonance (SPR) and autoimmune reactivity experiments. Finally, molecular dynamics (MD) simulation and binding free energy calculation (MM/PBSA) were performed on the wild-type CR3022 and its two double-site mutants to understand in more detail the contribution of these sites to the higher affinity. It was found that the binding affinity of the CR3022 antibody could be significantly enhanced more than ten times after the introduction of the S103F/Y mutation in HCDR–3 and the S33R mutation in LCDR–1. The additional hydrogen-bonding, hydrophobic interactions, as well as salt-bridges formed between the modified double-site mutated antibody and SARS-CoV-2 RBD were identified. The computational and experimental results clearly demonstrated that the affinity of the modified antibody has been greatly enhanced. This study indicates that CR3022 as a neutralizing antibody recognizing the conserved region of RBD against SARS-CoV with cross-reactivity with SARS-CoV-2, a different member in a large family of coronaviruses, could be improved by the computational and experimental approaches which provided insights for developing antibody drugs against SARS-CoV-2.     

Exposure marker discovery of di-2(propylheptyl) phthalate using ultra-performance liquid chromatography-mass spectrometry and a rat model

Di-(2-propylheptyl) phthalate (DPHP) is a plasticizer and has been suggested to be a subchronic toxicant in rats. DPHP has been approved to be used in food containers and handling by the U.S. Food and Drug Administration. The use of DPHP is still increasing, and the risk of human exposure to DPHP via food may be high. Exposure markers measured in human samples are commonly used to monitor human exposure levels. Ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) and a rat model were used to discover tentative DPHP exposure markers. DPHP and mono-(2-propylheptyl) phthalate (MPHP) were used as the precursors for calculating metabolite candidates using biotransformation mass changes of known enzymatic reactions. A rat model was designed to validate these metabolite candidates as tentative exposure markers. A total of 28 signals show dose–response relationships and these signals contain a few isomers. The chemical structures of 15 tentative exposure marker signals were speculated based on the product ion mass spectra from MS/MS analysis. These 15 signals included 7 chemical structures and some of them may be isomers. The different arrangement of the atoms in space of these isomers should be validated by standard compounds in the future studies. Among the 7 speculated chemical structures, 2 structures were novel tentative DPHP metabolites, and 5 structures have been previously reported in the literature. The results indicate that using UPLC-MS and a rat model can be used to identify tentative toxicant exposure markers.     

Facial intramuscular lipoma occurrence following topical cosmetic injection with a mixture of basic fibroblast growth factor: A report of two cases

Growth factors and cytokines control cell growth, proliferation and differentiation via a network of inter- and intracellular signalling pathways, and are involved in skin self-renewing and wound healing. In recent years, topical and injectable growth factors and cytokines have emerged as an intriguing therapeutic modality that can be harnessed for aesthetic purposes. However, very little data are available on their long-term safety and tolerability. In this report, we describe two cases of patients, who developed intramuscular lipoma of the chin following topical injection with a mixture of basic fibroblast growth factor as the main ingredients for chin augmentation. Biopsies in the two cases were performed at our department, and revealed intramuscular lipoma. Our report indicates that the topical injection of growth factors can lead to tumorigenesis, so health care providers need to be aware of its potential consequences.     

唑来膦酸治疗绝经后骨质疏松患者2年结果分析

目的 分析唑来膦酸（商品名：密固达）治疗绝经后骨质疏松患者2年后的疗效和不良反应.方法 统计我院2009~2012年中连续使用密固达治疗2年的绝经后骨质疏松患者47例的临床资料,分析治疗前后患者骨密度变化情况及不良反应发生率.结果 首次使用密固达患者中,出现发热3例（6.4%）,肌痛3例（6.4%）,发热合并肌痛9例（19.1%）.其中1例出现高热（最高体温40.5 ℃）;1例出现双侧大腿内侧出血点,1 w后消退;其余不良反应少见.第二次输注后出现发热5例（10.6%）,但无高热发生.左侧股骨颈骨密度、左侧全髋骨密度、L2~4椎体骨密度,治疗后与治疗前以及治疗第2年与第1年比较均明显升高（P＜0.05）.结论 密固达治疗绝经后骨质疏松对骨密度提升效果明显,且总体安全性好,主要不良反应为发热和肌痛.