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Supplemental material, sj-pdf-3-jcb-10.1177_0271678X231165653 for H3K9 trimethylation dictates neuronal ferroptosis through repressing Tfr1 by Ting Lan, Liye Hu, Tingting Sun, Xuechun Wang, Zhongnan Xiao, Danmin Shen, Weihua Wu, Zhaoli Luo, Chao Wei, Xiaotong Wang, Meng Liu, Yi Guo, Liyong Wang, Yamei Wang, Yabin Lu, Yan Yu, Fei Yang, Chenguang Zhang and Qian Li in Journal of Cerebral Blood Flow & Metabolism

Supplemental material, sj-pdf-4-jcb-10.1177_0271678X231165653 for H3K9 trimethylation dictates neuronal ferroptosis through repressing Tfr1 by Ting Lan, Liye Hu, Tingting Sun, Xuechun Wang, Zhongnan Xiao, Danmin Shen, Weihua Wu, Zhaoli Luo, Chao Wei, Xiaotong Wang, Meng Liu, Yi Guo, Liyong Wang, Yamei Wang, Yabin Lu, Yan Yu, Fei Yang, Chenguang Zhang and Qian Li in Journal of Cerebral Blood Flow & Metabolism

Background: Cuscutae Semen (CS) is reported to show a hepatoprotective effect. Chlorogenic acid, hyperoside and astragalin are three major biologically active components from CS. Objective: A sensitive method based on ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed and validated to quantify the three components in rat plasma and was successfully used to study pharmacokinetics in liver injury rats. Methods: Plasma samples were prepared with protein precipitation by acetonitrile. Chromatographic separation was achieved on ACQUITY-XBridge BEH C18 column with gradient elution using the mobile phase containing 0.05% formic acid in water (A) and acetonitrile (B). The three components were quantified using Electrospray Ionization (ESI) source in the negative multiple Reaction Monitoring (MRM) mode. Results: Calibration curves of each analyte showed good linearity with correlation coefficients over 0.99. Accuracies (RE%) and precisions (RSD%) were within 15%. The method was stable. Recovery of the target compounds in plasma samples ranged from 87.00% to 102.29%. No matrix effect was found to influence the quantitative method. Conclusion: The UPLC-MS/MS method met the acceptance criteria and was successfully applied to the simultaneous determination of chlorogenic acid, hyperoside and astragalin in rat plasma for the first time. It is suitable for pharmacokinetic application in liver injury rats. It provides the foundation for further development and utilization of the hepatoprotective effect of cuscutae semen.

A novel wearable human machine interface based on mechanomyogram (MMG) signals was presented in this study. A three-axis accelerometer was fixed to a customized watch strap to measure the MMG signals that were generated by the end of the extensor digitorum muscle. Eight gaming gestures, including clapping, index figure flicking, finger snapping, coin flipping, shooting, wrist extension, wrist flexion and fist-making, were identified in real time. This study extracted the features from both the time signals and the coefficients of the wavelet packet decomposition (WPD), and sequential forward selection (SFS) was used to identify the significant features to improve the classification accuracy and reduce the processing time. The performances of the classifiers such as the k-nearest neighbors (KNN), the support vector machine (SVM), linear discriminant analysis (LDA), and deep neural network (DNN) were compared. After testing the system on 35 subjects aged from 16 to 55 years old, the proposed system has advantages with respect to its convenient portability, stable signal acquisition, low power consumption, and high classification accuracy.

Background Primary lymphoma of the breast is rare, and primary diffuse large B cell lymphoma (DLBCL) of the breast is very rare. This study aimed to identify the clinicopathological characteristics and treatment associated with prognosis in patients with primary DLBCL of the breast. Material/Methods A retrospective study included the clinical and treatment data from 46 women with a histological diagnosis of primary DLBCL. Patients were staged using Ann Arbor staging criteria, overall survival (OS), progression-free survival (PFS), and the international prognostic index (IPI) scores were obtained. Laboratory finding included serum lactate dehydrogenase (LDH), and the immunohistochemistry findings were recorded. Results Patients (n=46), included stage I (n=18), stage II (n=18), stage III (n=3), and stage IV DLBCL (n=9). Treatment included chemotherapy with rituximab (n=16), and radiotherapy (n=12). The median follow-up time was 40.5 months, the 5-year OS rate was 36.2%, and the 5-year PFS rate was 29.1%. Univariate analysis showed that clinical stage, serum LDH, the IPI score, chemotherapy cycles >3, and Bcl-2 and Bcl-6 expression were correlated with the 5-year OS and PFS. Multivariate risk regression analysis showed that the number of chemotherapy cycles (>3) and Bcl-6 expression were independent prognostic factors in primary DLBCL of the breast (P3 cycles of chemotherapy and expression of Bcl-6 resulted in improved OS and PFS. Radiotherapy controlled local tumor recurrence but did not improve the OS and PFS. Rituximab did not improve patient survival.

Pneumatic artificial muscles (PAMs), as a type of actuators in soft robots, have been attracted in the past few decades. Because of their higher power density ratio and flexibility, PAMs are now widely used as a flexible actuator. When PAMs are applied to different robot systems, their dynamic output characteristics are influenced by the amplitude and frequency of input signals and external loads. The experimental results show that with the increase of input signals frequency, the width of PAMs dynamic hysteresis loop increases and the height decreases. Meanwhile, in order to verify that the fatigue life of the Mckibben PAMs made in the laboratory can meet the use requirements, this paper has completed the fatigue tests of them.

Zika virus (ZIKV) is a typical mosquito-borne flavivirus known to cause severe fetal microcephaly and adult Guillain-Barre syndrome. Currently, there are no specific drugs or licensed vaccines available for ZIKV infection, and further research is required to identify host cell proteins involved in the virus's life cycle. Viruses are known to use host cell membrane skeletal proteins, such as actin and spectrin, to complete cell entry, transportation, and release. Here, based on immunoprecipitation, the Axl and ZIKV envelope (E) protein were shown to interact with the cell membrane skeleton protein 4.1R. Furthermore, deletion of 4.1R significantly reduced virus titer and viral protein synthesis. Our study showed that 4.1R is an important host cell protein during ZIKV infection and may be involved in the process of viral entry into host cells.

Plants glycerol-3-phosphate dehydrogenase (GPDH) catalyzes the formation of glycerol-3-phosphate, and plays an essential role in glycerolipid metabolism and stress responses. In the present study, the knock-out mutants of cytosolic GPDH (AtGPDHc2) and wild-type Arabidopsis plants were treated with 0, 50, 100, and 150 mmol L−1 NaCl to reveal the effects of AtGPDHc2 deficiency on salinity stress responses. The fluctuation in redox status, reactive oxygen species (ROS) and antioxidant enzymes as well as the transcripts of genes involved in the relevant processes were measured. In the presence of 100 and 150 mmol L−1 NaCl treatments, AtGPDHc2-deficient plants exhibited a pronounced reduction in germination rate, fresh weight, root length, and overall biomass. Furthermore, loss of AtGPDHc2 resulted in a significant perturbation in cellular redox state (NADH/NAD+ and AsA/DHA) and consequent elevation of ROS and thiobarbituric acid-reactive substances (TBARS) content. The elevated ROS level triggered substantial increases in ROS-scavenging enzymes activities, and the up-regulated transcripts of the genes (CSD1, sAPX and PER33) encoding the antioxidant enzymes were also observed. In addition, the transcript levels of COX15, AOX1A and GLDH in gpdhc2 mutants decreased in comparison to wild-type plants, which demonstrated that the deficiency of AtGPDHc2 might also has impact on mitochondrial respiration under salt stress. Together, this work provides some new evidences on illustrating the roles of AtGPDHc2 playing in response to salinity stress by regulating cellular redox homeostasis, ROS metabolism and mitochondrial respiration.

Tangeretin is a polymethoxylated favone isolated from the peel of citrus fruits that possesses a broad spectrum of pharmacological functions. However, there has been limited information regarding tangeretin’s potential protective impact on the myocardium. As the myocardium is widely recognized as a potential target of oxidative stress induced by physical fatigue, this study aimed to assess tangeretin’s potential protective effects related to myocardial oxidative stress in fatigued mice. Using an intervention/control protocol on 5 groups of mice (N = 50; 1 control; 4 intervention; 4 week duration) with exhaustive swimming as the measured outcome, we found that tangeretin enhanced the activity of antioxidant enzymes, reduced the level of oxidative stress and injury, and protected myocardial morphology and ultrastructure. It was also found that tangeretin activated the Keap1/Nrf2 signaling axis to up-regulate the expression of phase II antioxidant and detoxifying enzymes. These findings suggest that tangeretin may impart beneficial effects by reducing physical fatigue-associated myocardial injury.

Both HIV and DENV are serious threats to human life, health and social economy today. So far, no vaccine for either HIV or DENV has been developed successfully. The research on anti-HIV or DENV drugs is still of great significance. In this study we developed a series of novel 2-Aryl-1H-pyrazole-S-DABOs with C6-strucutral optimizations as potent NNRTIs, among which, 8 compounds had low cytotoxicity and EC50 values in the range of 0.0508 ∼ 0.0966 μM, and their selectivity index was SI > 1415 ∼ 3940. In particular, two compounds 4a and 4b were identified to have good inhibitory effects on DENV of four serotypes. The EC50 of compound 4a and 4b against DENV-II (13.2 μM and 9.23 μM, respectively) were better than that of the positive control ribavirin (EC50 = 40.78 μM). In addition, the effect of C-6 substituents on the anti-HIV or anti-DENV activity of these compounds was also discussed.

BackgroundH7 subtype avian influenza has caused great concern in the global poultry industry and public health. The conventional serological subtype-specific diagnostics is implemented by hemagglutination inhibition (HI) assay despite lengthy operation time. In this study, an efficient, rapid and high-throughput competitive enzyme-linked immunosorbent assay (cELISA) was developed for detection of antibodies against H7 avian influenza virus (AIV) based on a novel monoclonal antibody specific to the hemagglutinin (HA) protein of H7 AIV.ResultsThe reaction parameters including antigen coating concentration, monoclonal antibody concentration and serum dilution ratio were optimized for H7 antibody detection. The specificity of the cELISA was tested using antisera against H1 ~ H9, H11 ~ H14 AIVs and other avian viruses. The selected cut-off values of inhibition rates for chicken, duck and peacock sera were 30.11, 26.85 and 45.66% by receiver-operating characteristic (ROC) curve analysis, respectively. With HI test as the reference method, the minimum detection limits for chicken, duck and peacock positive serum reached 20, 21and 2− 1HI titer, respectively. Compared to HI test, the diagnostic accuracy reached 100, 98.6, and 99.3% for chicken, duck and peacock by testing a total of 400 clinical serum samples, respectively.ConclusionsIn summary, the cELISA assay developed in this study provided a reliable, specific, sensitive and species-independent serological technique for rapid detection of H7 antibody, which was applicable for large-scale serological surveillance and vaccination efficacy evaluation programs.

Eight phenolic compounds including five glucuronides were isolated from the leaves of Cyclocarya paliurus (Batal.) Ijinskaja, which is used as a source of tea. Their structures were characterized by spectroscopic methods. The results of biological assay showed that compounds 1 and 5 had good activities against HIV-1 induced cytopathic effects in C8166 lymphocyte at non-cytotoxic concentrations, with EC50 values of 31.74 and 10.76 μM, and therapeutic index (TI) values of ＞13.02 and 25.46, respectively. Relationship between molecular structures and their bioactivities was discussed.

In order to discover and develop the new HIV-1 NNRTIs, a series of 5-alkyl-6-(benzo[d][1,3]dioxol-5-ylalkyl)-2-mercaptopyrimidin-4(3H)-ones was synthesized and screened for their in vitro cytotoxicity against HIV-1. Most of the compounds we synthetized showed high activity against wild-type HIV-1 strain (IIIB) while IC50 values are in the range of 0.06–12.95 μM. Among them, the most active HIV-1 inhibitor was compound 6-(benzo[d][1,3]dioxol-5-ylmethyl)-5-ethyl-2-((2-(4-hydroxyphenyl)-2-oxoethyl)thio)pyrimidin-4(3H)-one (5b), which exhibited similar HIV-1 inhibitory potency (IC50 = 0.06 μM, CC50 = 96.23 μM) compared with nevirapine (IC50 = 0.04 μM, CC50 >200 μM) and most of compounds exhibited submicromolar IC50 values indicating they were specific RT inhibitors. The compounds 5b, 6-(benzo[d] [1,3]dioxol-5-yl)-5-ethyl-2-((2-(4-hydroxyphenyl)-2-oxoethyl)thio)pyrimidin-4(3H)-one (5c) and 4-(2-((4-(benzo[d][1,3]dioxol-5-ylmethyl)-5-ethyl-6-oxo-1,6-dihydropyrimidin-2-yl)thio)acetyl)phenylbenzo[d][1,3]dioxole-5-carboxylate (5r) were selected for further study. It was found that all of them had little toxicity to peripheral blood mononuclear cell (PBMC), and had a good inhibitory effect on the replication of HIV-1 protease inhibitor resistant strains, fusion inhibitor resistant strains and nucleosides reverse transcriptase inhibitor resistant strains, as well as on clinical isolates. Besides, compound 5b and 5c showed inhibition of HIV-1 RT RNA-dependent DNA polymerization activity and DNA-dependent DNA polymerization activity, while compound 5r only showed inhibition of HIV DNA-dependent DNA polymerization activity, which was different from classical reverse transcriptase inhibitors. Our study which offered the preliminary structure-activity relationships and modeling studies of these new compounds has provided the valuable avenues for future molecular optimization.

Protein kinase is regarded as a potential target for anticancer therapeutics due to its relation to many diseases, in which more than one kinase participates in the cell signaling cascades. We herein demonstrate a multiplexed quantitative matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) approach to simultaneously assess the activity and inhibition of multiple protein kinases. In this design, substrate peptides phosphorylated by kinases of interest are specifically harvested by metal-organic framework (MOF)-templated porous CeO2 and consequently transformed to the dephosphorylated forms due to the phosphatase-like activity of CeO2, resulting in a unique quantitative MS signal with an enhanced intensity. Based on the peak area ratios of dephosphorylated variants of the phosphorylated product to respective deuterated internal standard, the activity and inhibition of each kinase can be independently profiled. In addition to the accurate characterization of protein kinase A (PKA) activity and inhibition induced by H-89, the multiplexing capability of the MS-based method allowed quantitative evaluation of the activity of Abl and Src, the two tightly associated kinases in the occurrence of chronic myeloid leukemia (CML) in a multiplexed format, exhibiting excellent orthogonality for the dual signal readout channels. Moreover, the inactivation of both Abl and Src by the inhibitor imatinib, dasatinib, and ponatinib was simultaneously traced, giving a whole picture of the competition behavior between the kinases for binding therapeutic molecules. This approach holds great promise in global investigation of kinase signal pathways and high-throughput screening of effective protein kinase inhibitors.

Background The human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) has always been a global health threat and leading cause of deaths. However, due to the emergence of drug-resistant HIV, an inevitable consequence of increasing use of antiretroviral drugs posed a major threat to antiretroviral therapy success. Objective The discovery of anti-HIV-1 agents will be used for the effective treatment of HIV/AIDS. Method In continuation of our program aimed to discover anti-HIV-1 agents, twelve matrine derivatives, such as 14-formyl-15-aryloxy/methoxymatrines (3a-j) and 14-aryloxymethylidenylmatrines (3k,l), were semi-prepared from matrine, and evaluated against HIV-1 IIIB replication in acutely infected C8166 cells in vitro. Results Among them, compound 3j showed the most potent anti-HIV-1 activity with EC50 and therapeutic index (TI) values of 1.79 µg/mL, and 98.2, respectively. Conclusion It has been demonstrated that the positions of methyl on the phenyl ring and 14- formylmatrine-15-oxy on the naphthyl ring were very important for the activity. It will lay the foundation for further structural modification and application of matrines as HIV-1 inhibitors.

Many human proteins have the potential to be developed as therapeutic agents. However, side effects caused by direct administration of natural proteins have significantly slowed expansion of protein therapeutics into the clinic. Post-translational modifications (PTMs) can improve protein properties, but because of significant knowledge gaps, we are considerably limited in our ability to apply PTMs to generate better protein therapeutics. Here, we seek to fill the gaps by studying the PTMs of a small representative chemotactic cytokine, RANTES. RANTES can inhibit HIV-1 infection by competing with it for binding to receptor CCR5 and stimulating CCR5 endocytosis. Unfortunately, RANTES can induce strong signaling, leading to severe inflammatory side effects. We apply a chemical biology approach to explore the potential of post-translationally modified RANTES as safe inhibitors of HIV-1 infection. We synthesized and systematically tested a library of RANTES isoforms for their ability to inhibit inflammatory signaling and prevent HIV-1 infection of primary human cells. Through this research, we revealed that most of the glycosylated variants have decreased inflammation-associated properties and identified one particular glyco variant, a truncated RANTES containing a Galβ1-3GalNAc disaccharide α-linked to Ser4, which stands out as having the best overall properties: relatively high HIV-1 inhibition potency but also weak inflammatory properties. Moreover, our results provided a structural basis for the observed changes in the properties of RANTES. Taken together, this work highlights the potential importance of glycosylation as an alternative strategy for developing CCR5 inhibitors to treat HIV-1 infection and, more generally, for reducing or eliminating unwanted properties of therapeutic proteins.

Pathogenesis-related (PR) proteins are generally involved in the defense of plants and are important contributors in the disease resistance of plants. Among the 17 PRs that are currently recognized, the PR4 family of proteins is divided into two classes and features a conserved barwin domain. In this study, we isolated two Class II PR4s from the oriental hybrid lily cultivar Sorbonne using the rapid amplification of the cDNA ends (RACE) method, and designated these two PR4s LhSorPR4a and LhSorPR4b. LhSorPR4a and LhSorPR4b were 627 and 617 bp in length, respectively, and encoded two corresponding PR4s of 141 and 143 amino acids. These deciphered LhSorPR4a and LhSorPR4b protein sequences shared a sequence similarity of 90.7%, but their theoretical isoelectric points were distinctively different (7.74 and 4.08, respectively). The three-dimensional structures of LhSorPR4a and LhSorPR4b predicted by homology modeling showed high similarity to their corresponding papaya barwin-like protein template. Analysis of expression by qPCR revealed that both LhSorPR4a and LhSorPR4b were responsive to methyl jasmonate and ethephon treatments. The LhSorPR4b expression was also significantly induced by sodium salicylate (SS); however, LhSorPR4a was unresponsive to the SS treatment. Both LhSorPR4a and LhSorPR4b were expressed in Escherichia coli (E. coli) and successfully purified. The PR4s characterized in this study (LhSorPR4a and LhSorPR4b) are the first two PR4 family genes isolated from the Lilium genus, and they could therefore play an important role in lily disease resistance.

Long noncoding RNAs (lncRNAs) mediate important epigenetic regulation in a wide range of biological processes and diseases. We applied comprehensive analyses of RNA-seq and CAGE-seq (cap analysis of gene expression and sequencing) to characterize the dynamic changes in lncRNA expression in rhesus macaque (Macaca mulatta) brain in four representative age groups. We identified 18 anatomically diverse lncRNA modules and 14 mRNA modules representing spatial, age, and sex specificities. Spatiotemporal- and sex-biased changes in lncRNA expression were generally higher than those observed in mRNA expression. A negative correlation between lncRNA and mRNA expression in cerebral cortex was observed and functionally validated. Our findings offer a fresh insight into spatial-, age-, and sex-biased changes in lncRNA expression in macaque brain and suggest that the changes represent a previously unappreciated regulatory system that potentially contributes to brain development and aging.

Background and objective Probiotic bacteria can induce immune regulation or immune tolerance in patients with allergic diseases, but the underlying mechanisms are still unclear. There has been a growing interest in the use of beneficial bacteria for allergic diseases recently. This study aimed at exploring whether Clostridium butyricum CGMCC0313-1 (C. butyricum) can reduce ovalbumin (OVA)-induced allergic airway inflammation in a mouse model. Methods Mouse model of allergic airway inflammation induced via OVA was used in this study. C. butyricum was administered daily by the oral route during or after the sensitization. Airway function, pulmonary airway inflammation, mast cell degranulation, T helper (Th)-specific and anti-inflammatory cytokines, OVA-specific Ig, matrix metalloproteinase 9 (MMP-9) and histopathological alterations were examined. Results C. butyricum significantly reduced lung resistance in the asthmatic mice. Pulmonary airway inflammation, mast cell degranulation, airway remodelling and the expression of OVA-specific IgE/G1 were suppressed by oral C. butyricum. It also reversed the imbalance of Th1/Th2 and increased the anti-inflammatory cytokine IL-10. Conclusion C. butyricum reduces OVA-induced allergic airway inflammation in mice and might be an additional or supplementary therapy for allergic asthma.

The present study investigated the efficiency of the use of chitosan nanoparticles containing plasmid‑bone morphogenetic protein 2 (pBMP2) sequences (CNPBs) to induce the differentiation of bone marrow stem cells (BMSCs) into osteoblast‑like cells that may be able to promote ectopic bone formation. pBMP2s were constructed, and chitosan nanoparticles were incubated with 50, 100 or 200 µg/ml pBMP2. BMSCs were collected from the tibiae and femurs of 6‑week old rats, cultured and treated with the CNPBs or 200 µg/ml pBMP2 as a positive control. Transfection efficiency was confirmed using the green fluorescent protein assay. Histological staining methods, including alkaline phosphatase, Wright's and von Kasso staining, were used to identify features of osteoblast‑like cells differentiated from BMSCs. Expression levels of the markers of osteoblasts, such as alkaline phosphatase, osteoprotegerin, osteocalcin and osteopontin, were determined to verify the differentiation of BMSCs into osteoblast‑like cells. Ectopic bone formation was observed following the integration of polyglycolic acid (PGA) scaffolds with CNPBs and BMSCs, which were implanted into the dorsal muscles of Sprague‑Dawley rats. Exposure to CNPBs led to the transfection of BMSCs with BMP2. The transfected BMSCs possessed the characteristic phenotypes of osteoblasts. The expression levels of alkaline phosphatase, osteoprotegerin, osteocalcin and osteopontin were significantly higher in the transfected cells compared with the control group, particularly the CBP200 group. PGA scaffolds integrated with BMSCs and CNPBs induced ectopic bone formation, as changes in the morphology of cells were observed using histological staining. Therefore, CNPBs may be a promising method of promoting the formation of novel bone tissue.

Antiretroviral therapy (ART) can effectively suppress human immunodeficiency virus type 1 (HIV-1) replication and limit the disease progression. However, ART is unable to eradicate the virus, and the requirement for lifelong treatment may have side effects and poses a risk of the development of resistance. New approaches to prevent and treat HIV-1 infection should therefore be developed. HIV-1 capsid (CA) protein is an unexploited but attractive target for antiviral drug development. The hydrophobic cavity of C-terminal domain of CA (CA CTD) has been validated as a potential target for antiviral drugs. Binding of compounds to this conserved nonpolar groove in CA CTD allosterically disrupts the CA assembly. In the study described here, we screened 2,080 natural products to identify potential antiviral agents for further development to combat HIV-1 infection. From the primary screen at a fixed concentration of 50 µM, 16 compounds were found to be effective against this target. Then, 6 compounds observed in the primary screen were confirmed in dose-response experiments. Finally, the 6 selected compounds were tested against HIV-1-induced cytopathic effects. Two compounds showed inhibitory effect on HIV-1 replication, and the most active one, rubranol, inhibited viral replication at a moderate micromolar concentration (EC50 = 15.85 μM). The binding mode of rubranol and hirsutanonol to CA CTD were analyzed by molecular docking respectively, providing insight for design of drug targeting HIV-1 capsid. The research reports, for the first time, identification of natural products that showed potential as anti-HIV-1 agents by targeting conserved hydrophobic cavity of HIV-1 CA CTD.

Background Currently, over 20 genes have been defined that can confer susceptibility for high-risk breast cancer. Although research has proved the utility of multiple-gene sequencing in the assessment of breast cancer risk, there is little data from China patients. Here, we use a multiple-gene sequencing panel to identify the variant spectrum in Chinese high-risk breast cancer subjects. Findings A total of 829 Chinese high-risk breast cancer patients participated in the research. The coding regions of 115 hereditary cancer susceptibility genes were captured and sequenced using a next generation sequencing platform from Complete Genomics and Illumina. Single Nucleotide Polymorphism and small Insertion/deletion variants were defined and analyzed. We developed a semi-automatic interpretation pipeline which enables accurate variants classification. In total, 193 pathogenic variants were identified in 45 genes from 177 patients. The pathogenic variant carrier rate is 21.4%: with 10.5% patients carrying a BRCA1 or BRCA2 mutation only, 10.0% of patients carried non-BRCA gene mutations only, while 1.0% of patients carried both a BRCA1/2 and a non-BRCA gene mutation. Variants of uncertain significance (VUS) totaling 2632 were identified in 115 genes from 787 of 829 patients: 82.5% patients carried more than one VUS, and only 5.1% patients did not carry any VUS. Families carrying pathogenic variants were tracked and adenoma was founded in three of them. Conclusions Our data provide a comprehensive analysis of potential susceptibility variations of high-risk for breast cancer in a Chinese population. This data will be useful for the comparison of the susceptibility variation spectrum between different populations and to discover potential pathogenic variants to improve the prevention and treatment of high-risk breast cancer.

OBJECTIVE To evaluate the safety and efficacy of Tongkuaixiao ointment (TKXO) in treating moderate-to-severe cancer induced somatalgia. METHODS Totally 130 patients with moderate-to-severe cancer induced somatalgia were randomly divided into a TKXO group and a control group. The patients were treated with either TKXO applied externally or placebo, with opioid analgesics orally at the same time. Observation parameters were included numerical rating scale (NRS) scores, analgesic efficacy, initiation effective time, persistent analgesic time, equivalent morphine dose, National Comprehensive Cancer Network (NCCN) grade in Impact of Pain Measurement Scores, and safety and satisfaction extent investigation. RESULTS NRS scores and NCCN grade in Impact of Pain Measurement Scores decreased significantly after 5-days’ treatment in the two groups (P < 0.0001). Compared to the control group, initiation effective time was significantly shorter (P < 0.05) and persistent analgesic time was significantly longer (P < 0.01), equivalent oral morphine doses of the first day and the whole treatment course were significantly decreased in the TKXO treatment group (P < 0.01 or P < 0.05). No obvious adverse effects were found in the TKXO group. CONCLUSION TKXO combined with opioid analgesics possesses the advantages of high efficacy, fast action, long persistent action, safety and convenience in use, and it can reduce the dose of opioid.

In a previous study the small molecule SJP-L-5 that inhibits HIV replication, has been shown to block uncoating of the viral capsid. Continued study showed that SJP-L-5 might hinder HIV capsid uncoating by blocking the completion of reverse transcription. However, to date, the mechanism has not been fully elucidated. Here, the effects of SJP-L-5 for reverse transcription were explored via quantitative PCR, DIG-labelled ELISA, fluorescent resonance energy transfer, and Southern blot assays. We also analyzed the resistance profile of this compound against reverse transcriptase. Our results show that SJP-L-5 preferentially inhibits PPT primed plus-strand DNA synthesis (EC50 = 13.4 ± 3.0 μM) over RNA primed minus-strand DNA synthesis (EC50 > 3,646 μM), resulting in formation of five segmented plus-strand DNA and loss of HIV DNA flap, suggesting failure of both nuclear import and integration. Moreover, resistance study evidenced that SJP-L-5 requires the amino acid residues Val108 and Tyr181 to exert an inhibitory effect. These results indicate SJP-L-5 as a new non-nucleoside reverse transcriptase inhibitor that inhibits HIV-1 polypurine tract primed plus-strand DNA synthesis, initiating HIV-1 down-stream plus-strand DNA synthesis at multiple sites under drug pressure.

Background. Malignant pleural effusion (MPE) is a common complication in most malignancies. Despite its frequent occurrence, current knowledge of MPE remains limited and the effect of the management is still unsatisfying. Traditional Chinese medicine (TCM) external treatment has unique advantages, such as quicker efficacy and fewer side effects. Objective. To observe the effects and safety of Kang’ai Xiaoshui ointment (TCM herbal ointment) in MPE. Design. This was a placebo-controlled double-blinded randomized study. A total of 80 patients were enrolled, of which 72 were randomized to receive Kang’ai Xiaoshui ointment or placebo at an allocation ratio of 1:1. Kang’ai Xiaoshui ointment or placebo was applied on the thorax wall for 8 hours daily. The intervention lasted 2 weeks. Kang’ai Xiaoshui ointment consisted of Astragalus membranaces （黄芪）, Semen pharbitidis （牵牛子）, Cassia twig （桂枝）, Pericarpium arecae （大腹皮）, Curcuma zedoary （莪术）, Borneol （冰片), and other substances. In both groups, diuresis and drainages were used as needed. Outcomes covered the quantity of pleural effusion evaluation, TCM Symptom Scale, Karnofsky Performance Scale, and safety indicators such as routine blood test, blood biochemistry test, and response table of skin irritation. Results. Of 72 patients randomized to receive Kang’ai Xiaoshui ointment or placebo along with symptomatic treatment, the response rate was documented as 42.4% for the treatment group and 25.0% for the placebo group ( P = .138). As for the TCM symptom scale, the treatment group showed improvement in chest distress ( P = .003), fullness and distention ( P = .042), shortness of breath ( P < .001), no statistical significance in palpitation ( P = .237), and pain ( P = .063), whereas the placebo group did not show statistical significance in any of the 5 symptoms. Major adverse events related to the treatment, mainly skin irritation, were distributed equally. Conclusions. Kang’ai Xiaoshui ointment showed a potential of reducing MPE, and it could alleviate symptoms of dyspnea. Thus, it may be appropriate as a supplementary intervention for MPE. There were some flaws in the study design. A larger scale and better designed trial is advocated.

Searching for more safe and effective agents for HIV treatments is still an urgent topic worldwide. Based on our continuous modifications on the benzophenone derivatives as HIV-1 reverse transcriptase (RT) inhibitors, a new template bearing N-phenylbenzenesulfonamide (PBSA) structure was designed to enhance the interactions with HIV-1 RT. In this manuscript, a series of PBSA derivatives were synthesized and evaluated for their anti-HIV-1 activity. The preliminary test showed that these compounds were potent to inhibit wild-type HIV-1 with EC50 values ranging of 0.105–14.531 μmol/L. In particular, compound 13f not only has high anti-HIV-1 activity (0.108 μmol/L), but also possesses low toxicity with a TI value of 1816.6. Furthermore, the major interactions of the inhibitor 13f with HIV-1 RT were also investigated using the molecular modelling. Our discovered structure-activity relationships (SARs) of these analogues may serve as an important clue for further optimizations.

A series of small-molecule Plk1 inhibitors targeting the substrate-binding pocket were designed through rational drug design for the first time. The designed compounds were synthesized and their activities were evaluated in vitro. Some of the targeted compounds showed potent Plk1 inhibitory activities and anti-proliferative characters. Particularly, 5i showed Plk1 inhibitory activity with an IC50 value of 0.68 µM. Compound 5i also showed cell growth inhibitory activity on HeLa cells with an IC50 value of 0.51 µM, which is about four times more potent compared to thymoquinone. The mechanism of action suggested that 5i was an ATP-independent and substrate-dependent Plk1 inhibitor. Compound 5i demonstrated excellent Plk1 inhibitory selectivity against Plk2, Plk3, and five serine/threonine and tyrosine kinases. Our discovery and structure–activity relationship study may provide useful lead compounds for further optimization of non-ATP-competitive Plk1 inhibitors.