Identification of FTY720 and COH29 as novel topoisomerase I catalytic inhibitors by experimental and computational studies.

The development of novel topoisomerase I (TOP1) inhibitors is crucial for overcoming the drawbacks and limitations of current TOP1 poisons. Here, we identified two potential TOP1 inhibitors, namely, FTY720 (a sphingosine 1-phosphate antagonist) and COH29 (a ribonucleotide reductase inhibitor), through experimental screening of known active compounds. Biological experiments verified that FTY720 and COH29 were nonintercalative TOP1 catalytic inhibitors that did not induce the formation of DNA-TOP1 covalent complexes. Molecular docking revealed that FTY720 and COH29 interacted favorably with TOP1. Molecular dynamics simulations revealed that FTY720 and COH29 could affect the catalytic domain of TOP1, thus resulting in altered DNA-binding cavity size. The alanine scanning and interaction entropy identified Arg536 as a hotspot residue. In addition, the bioinformatics analysis predicted that FTY720 and COH29 could be effective in treating malignant breast tumors. Biological experiments verified their antitumor activities using MCF-7 breast cancer cells. Their combinatory effects with TOP1 poisons were also investigated. Further, FTY720 and COH29 were found to cause less DNA damage compared with TOP1 poisons. The findings provide reliable lead compounds for the development of novel TOP1 catalytic inhibitors and offer new insights into the potential clinical applications of FTY720 and COH29 in targeting TOP1.

Assembling High-Quality Lymph Node Clinical Target Volumes for Cervical Cancer Radiotherapy Using a Deep Learning-Based Approach.

AIM: The study aimed to explore an approach for accurately assembling high-quality lymph node clinical target volumes (CTV) on CT images in cervical cancer radiotherapy with the encoder-decoder 3D network. METHODS: 216 cases of CT images treated at our center between 2017 and 2020 were included as a sample, which were divided into two cohorts, including 152 cases and 64 controls, respectively. Para-aortic lymph node, common iliac, external iliac, internal iliac, obturator, presacral, and groin nodal regions were delineated as sub-CTV manually in the cohort including 152 cases. Then, the 152 cases were randomly divided into training (96 cases), validation (36 cases), and test (20 cases) groups for the training process. Each structure was individually trained and optimized through a deep learning model. An additional 64 cases with 6 different clinical conditions were taken as examples to verify the feasibility of CTV generation based on our model. Dice similarity coefficient (DSC) and Hausdorff distance (HD) metrics were both used for quantitative evaluation. RESULTS: Comparing auto-segmentation results to ground truth, the mean DSC value/HD was 0.838/7.7mm, 0.853/4.7mm, 0.855/4.7mm, 0.844/4.7mm, 0.784/5.2mm, 0.826/4.8mm and 0.874/4.8mm for CTV_PAN, CTV_common iliac, CTV_internal iliac, CTV_external iliac, CTV_obturator, CTV_presacral, and CTV_groin, respectively. The similarity comparison results of six different clinical situations were 0.877/4.4mm, 0.879/4.6mm, 0.881/4.2mm, 0.882/4.3mm, 0.872/6.0mm, and 0.875/4.9mm for DSC value/HD, respectively. CONCLUSION: We have developed a deep learning-based approach to segmenting lymph node sub-regions automatically and assembling high-quality CTVs according to clinical needs in cervical cancer radiotherapy. This work can increase the efficiency of the process of cervical cancer detection and treatment.

Identification of N-(3-(methyl(3-(orotic amido)propyl)amino)propyl) oleanolamide as a novel topoisomerase I catalytic inhibitor by rational design, molecular dynamics simulation, and biological evaluation.

DNA topoisomerase I (TOP1) catalytic inhibitors are a promising class of antitumor agents. Oleanolic acid derivatives are potential TOP1 catalytic inhibitors. However, their inhibitory activity still needs to be enhanced, and the stability and hotspot residue sites of their interaction with TOP1 remain to be elucidated. Herein, a novel oleanolic acid derivative, OA4 (N-(3-(methyl(3-(orotic amido)propyl)amino)propyl)oleanolamide), was identified by rational design. Subsequently, molecular dynamics simulations were performed to explore the stability and conformational dynamics of the TOP1-OA4 complex. The molecular mechanics/generalized Born surface area method calculated the binding free energy and predicted Arg488, Ile535, and His632 to be hotspot residues. Biological experiments verified that OA4 is a nonintercalative TOP1 catalytic inhibitor. OA4 exhibits better proliferation inhibitory activity against tumor cells than normal cells. Furthermore, OA4 can induce apoptosis and effectively suppress the proliferation and migration of cancer cells. This work provides new insights for the development of novel TOP1 catalytic inhibitors.

Soluble CD4 effectively prevents excessive TLR activation of resident macrophages in the onset of sepsis.

T lymphopenia, occurring in the early phase of sepsis in response to systemic inflammation, is commonly associated with morbidity and mortality of septic infections. We have previously shown that a sufficient number of T cells is required to constrain Toll-like receptors (TLRs) mediated hyperinflammation. However, the underlying mechanisms remains unsolved. Herein, we unveil that CD4+ T cells engage with MHC II of macrophages to downregulate TLR pro-inflammatory signaling. We show further that the direct contact between CD4 molecule of CD4+ T cells or the ectodomain of CD4 (soluble CD4, sCD4), and MHC II of resident macrophages is necessary and sufficient to prevent TLR4 overactivation in LPS and cecal ligation puncture (CLP) sepsis. sCD4 serum concentrations increase after the onset of LPS sepsis, suggesting its compensatory inhibitive effects on hyperinflammation. sCD4 engagement enables the cytoplasmic domain of MHC II to recruit and activate STING and SHP2, which inhibits IRAK1/Erk and TRAF6/NF-κB activation required for TLR4 inflammation. Furthermore, sCD4 subverts pro-inflammatory plasma membrane anchorage of TLR4 by disruption of MHC II-TLR4 raft domains that promotes MHC II endocytosis. Finally, sCD4/MHCII reversal signaling specifically interferes with TLR4 but not TNFR hyperinflammation, and independent of the inhibitive signaling of CD40 ligand of CD4+ cells on macrophages. Therefore, a sufficient amount of soluble CD4 protein can prevent excessive inflammatory activation of macrophages via alternation of MHC II-TLR signaling complex, that might benefit for a new paradigm of preventive treatment of sepsis.

[Study on the sensitivity of a volumetric modulated arc therapy plan verification equipment on multi-leaf collimator opening and closing errors and its gamma pass rate limit].

To investigate the γ pass rate limit of plan verification equipment for volumetric modulated arc therapy (VMAT) plan verification and its sensitivity on the opening and closing errors of multi-leaf collimator (MLC), 50 cases of nasopharyngeal carcinoma VMAT plan with clockwise and counterclockwise full arcs were randomly selected. Eight kinds of MLC opening and closing errors were introduced in 10 cases of them, and 80 plans with errors were generated. Firstly, the plan verification was conducted in the form of field-by-field measurement and true composite measurement. The γ analysis with the criteria of 3% dose difference, distance to agreement of 2 mm, 10% dose threshold, and absolute dose global normalized conditions were performed for these fields. Then gradient analysis was used to investigate the sensitivity of field-by-field measurement and true composite measurement on MLC opening and closing errors, and the receiver operating characteristic curve (ROC) was used to investigate the optimal threshold of γ pass rate for identifying errors. Tolerance limits and action limits for γ pass rates were calculated using statistical process control (SPC) method for another 40 cases. The error identification ability using the tolerance limit calculated by SPC method and the universal tolerance limit (95%) were compared with using the optimal threshold of ROC. The results show that for the true composite measurement, the clockwise arc and the counterclockwise arc, the descent gradients of the γ passing rate with per millimeter MLC opening error are 10.61%, 7.62% and 6.66%, respectively, and the descent gradients with per millimeter MLC closing error are 9.75%, 7.36% and 6.37%, respectively. The optimal thresholds obtained by the ROC method are 99.35%, 97.95% and 98.25%, respectively, and the tolerance limits obtained by the SPC method are 98.98%, 97.74% and 98.62%, respectively. The tolerance limit calculated by SPC method is close to the optimal threshold of ROC, both of which could identify all errors of ±2 mm, while the universal tolerance limit can only partially identify them, indicating that the universal tolerance limit is not sensitive on some large errors. Therefore, considering the factors such as ease of use and accuracy, it is suggested to use the true composite measurement in clinical practice, and to formulate tolerance limits and action limits suitable for the actual process of the institution based on the SPC method. In conclusion, it is expected that the results of this study can provide some references for institutions to optimize the radiotherapy plan verification process, set appropriate pass rate limit, and promote the standardization of plan verification.

Activating Blood Circulation, Anti-Inflammatory and Diuretic Effects of Leonurus japonicus Extract on a Rat Model of Trauma Blood Stasis and Its Phytochemical Profiling.

Leonurus japonicus Houtt. has been traditionally used to treat many ailments. This study evaluated the activating blood circulation, anti-inflammatory, and diuretic effects of L. japonicus extract (LJ) and identified its phytochemicals. In this work, the phytochemicals in LJ were identified using liquid chromatography mass spectrometry. Rats were randomly assigned to three groups (n=8): Control group was treated with saline, while the Model group (saline) and LJ group (426 mg/kg) had induced traumatic injury. All rats were treated with once by daily oral gavage for one week. The biochemical indices and protein expression were measured. Herein, 79 constituents were identified in LJ, which were effective in elevating body weight, food consumption, water intake, and urinary excretion volume, as well as in ameliorating traumatic muscle tissues in model rats. In addition, LJ prominently decreased the contents of plasma viscosity, platelet aggregation rate, thrombin time, prothrombin time, activated partial thromboplastin time, fibrinogen, thromboxane B2 (TXB2), TXB2/6-keto-prostaglandin F1α (6-keto-PGF1α), urokinase-type plasminogen activator (u-PA), plasminogen activator inhibitor 1 (PAI-1), PAI-1/tissue-type PA (t-PA), and PAI-1/u-PA, while significantly increasing antithrombin III, 6-keto-PGF1α, and t-PA contents. Furthermore, LJ notably inhibited tumor necrosis factor alpha, interleukin 6 (IL-6), IL-8, angiotensin II, antidiuretic hormone, aldosterone, aquaporin 1 (AQP1), AQP2, and AQP3 levels, and markedly elevating IL-10 and natriuretic peptide levels. Finally, LJ markedly reduced the protein expression of AQP1, AQP2, and AQP3 compared to the model group. Collectively, LJ possessed prominent activating blood circulation, anti-inflammatory, and diuretic effects, thus supporting the clinical application of L. japonicus.

Revealing the Active Constituents and Mechanisms of Semiliquidambar cathayensis Chang Roots against Rheumatoid Arthritis through Network Pharmacology, Molecular Docking, and in Vivo Experiment.

Semiliquidambar cathayensis Chang roots (SC) are traditional Chinese medicine for treating rheumatoid arthritis (RA). However, the effect and potential mechanism of SC remain unclear. This study aims to reveal the anti-RA constituents and mechanisms of SC based on network pharmacology, molecular docking, and adjuvant-induced arthritis (AIA) model rat experiment. In this work, 9 potential active constituents, including kaempferol, quercetin, naringenin, paeoniflorin, catechin, fraxin, gentianin, hesperetin, and ellagic acid 3,3',4-trimethyl ether, in SC crossed 65 target genes of RA. In addition, 28 core targets were enriched in inflammation and others, among which interleukin-17 (IL-17) and tumor necrosis factor (TNF) were the major targets. The binding of bio-constituents with IL-17 and TNF were performed using molecular docking. Rat experiment demonstrated that the extract of SC restored body weight loss, reduced arthritis score and the indices of thymus and spleen, alleviated ankle joint histopathology, decreased the levels of rheumatoid factor (RF), C-reactive protein (CRP), IL-17, TNF-α, IL-1ß, IL-6, cyclooxygenase-2 (COX-2), 5-lipoxygenase (5-LOX), and matrix metalloproteinase-2 (MMP-2), whereas elevated the levels of IL-4 and IL-10. Collectively, it was the first time to comprehensively reveal the anti-RA efficacy and mechanism of SC via suppressing the inflammatory pathway based on network pharmacology, molecular docking, and experimental verification, which provide chemical and pharmacological evidences for the clinical application of SC.

Sensitivity of Three Patient-Specific Quality Assurance Systems to MLC Aperture Errors With Volumetric Modulated Arc Therapy.

Purpose: To compare the sensitivity of ArcCHECK (AC), portal dosimetry (PD), and an in-house logfile-based system (LF) to multileaf collimators (MLC) aperture errors and the ability to identify these errors. Methods and Materials: For 12 retrospective original head and neck volumetric modulated arc therapy (VMAT) plans, MLC aperture errors of ± 0.4mm, ± 1.2mm, ± 2mm, and ± 3mm were introduced for each plan, resulting in 96 plans with errors. AC, PD, and LF were used for the gamma evaluation at 3%/3mm, 3%/2mm, and 2%/2mm criteria. Gradient analysis was used to evaluate the sensitivity to MLC aperture errors. The area under the curve (AUC) obtained from the receiver operating characteristic (ROC) curve was used to evaluate the ability to identify MLC aperture errors and dose errors, and the optimal cut-off value to identify the error was obtained. Results: The gamma pass rate (%GP) of LF had the smallest descent gradient as the MLC error increases in any case. The descent gradient of PD was larger than AC, except for the case at the 2%/2mm criteria. For the 3%/3mm criteria, the MLC aperture errors that can be perfectly identified by AC, PD, and LF were ± 3mm, ± 2mm, and ± 1.2mm, respectively, and the average percent dose error (%DEs) of dose metrics in targets that can be perfectly identified were 4% to 5%, 3% to 4%, and 2% to 3%, respectively. For the 3%/2mm criteria, the errors that AC, PD, and LF can perfectly identify were the same as the 3%/3mm criteria. For the 2%/2mm criteria, AC can perfectly identify the MLC error of ± 2mm and the %DE of 3% to 4%. PD and LF can identify the MLC error of ± 1.2mm and the %DE of 2% to 3%. Conclusion: Different patient-specific quality assurance (PSQA) systems have different sensitivity and recognition abilities to MLC aperture errors. Institutions should formulate their own customized %GP limits based on their PSQA process through ROC or other methods.

Case Report: Successful ABO-Incompatible Deceased Donor Kidney Transplantation in an Infant Without Pre-transplant Immunological Treatment.

ABO blood group antibodies have not been generated or are at low titer during early infancy. Therefore, in theory, ABO-incompatible kidney transplantation (ABOi KT) may be successfully achieved in small infants without any pre-transplant treatment. We report here the first ABO-incompatible deceased donor kidney transplantation (ABOi DDKT) in an infant. The recipient infant was ABO blood group O, and the donor group A. The recipient was diagnosed with a Wilms tumor gene 1 (WT1) mutation and had received peritoneal dialysis for 4 months prior to transplant. At 7 months and 27 days of age, the infant underwent bilateral native nephrectomy and single-kidney transplantation from a 3-year-old brain-dead donor. No pre- or post-transplantation antibody removal treatment was performed, since the recipient's anti-iso-hemagglutinin-A Ig-M/G antibody titers were both low (1:2) before transplantation and have remained at low levels or undetectable to date. At 11 months post-transplant, the recipient is at home, thriving, with normal development and graft function. This outcome suggests that ABOi DDKT without antibody removal preparatory treatment is feasible in small infants, providing a new option for kidney transplantation in this age range.

Improved curved-boundary scheme for lattice Boltzmann simulation of microscale gas flow with second-order slip condition.

An improved curved-boundary scheme with second-order velocity slip condition for multiple-relaxation-time-lattice Boltzmann (MRT-LB) simulation of microgas flow is proposed. The proposed interpolation bounce-back (IBB)-explicit counter-extrapolation (ECE) scheme adopts the IBB method to describe the curved boundary, while the ECE method is employed to predict the slip velocity on gas-solid interface. To incorporate the effect of second-order velocity slip term and the influence of boundary curvature, a slip velocity model is also derived, from which the gas slip velocity is captured by the ECE discretization method. The influence of fictitious slip velocity can be eliminated by adopting the present ECE method, and the influence of actual offset between the lattice node and the physical boundary can be well considered by the IBB method. The proposed IBB-ECE boundary scheme is then implemented with the MRT-LB model and tested by simulations of force-driven gas flow in horizontal (inclined) microchannel, gas flow around a micro-cylinder, and Couette flow between two micro-cylinders. Numerical results show that the proposed IBB-ECE scheme improves the computational accuracy of gas slip flow (0.001

SARS-CoV-2 helicase NSP13 hijacks the host protein EWSR1 to promote viral replication by enhancing RNA unwinding activity.

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in December 2019 and has led to a global coronavirus disease 2019 (COVID-19) pandemic. Currently, incomplete understanding of how SARS-CoV-2 arrogates the host cell to establish its life cycle has led to slow progress in the development of effective drugs. Results: In this study, we found that SARS-CoV-2 hijacks the host protein EWSR1 (Ewing Sarcoma breakpoint region 1/EWS RNA binding protein 1) to promote the activity of its helicase NSP13 to facilitate viral propagation. NSP13 is highly conserved among coronaviruses and is crucial for virus replication, providing chemical energy to unwind viral RNA replication intermediates. Treatment with different SARS-CoV-2 NSP13 inhibitors in multiple cell lines infected with SARS-CoV-2 effectively suppressed SARS-CoV-2 infection. Using affinity-purification mass spectrometry, the RNA binding protein EWSR1 was then identified as a potent host factor that physically associated with NSP13. Furthermore, silencing EWSR1 dramatically reduced virus replication at both viral RNA and protein levels. Mechanistically, EWSR1 was found to bind to the NTPase domain of NSP13 and potentially enhance its dsRNA unwinding ability. Conclusions: Our results pinpoint EWSR1 as a novel host factor for NSP13 that could potentially be used for drug repurposing as a therapeutic target for COVID-19.

Quantitative Comparisons of Deep-learning-based and Atlas-based Auto- segmentation of the Intermediate Risk Clinical Target Volume for Nasopharyngeal Carcinoma.

BACKGROUND: Manual segment target volumes were time-consuming and inter-observer variability couldn't be avoided. With the development of computer science, auto-segmentation had the potential to solve this problem. OBJECTIVE: To evaluate the accuracy and stability of Atlas-based and deep-learning-based auto-segmentation of the intermediate risk clinical target volume, composed of CTV2 and CTVnd, for nasopharyngeal carcinoma quantitatively. METHODS AND MATERIALS: A cascade-deep-residual neural network was constructed to automatically segment CTV2 and CTVnd by deep learning method. Meanwhile, a commercially available software was used to automatically segment the same regions by Atlas-based method. The datasets included contrast computed tomography scans from 102 patients. For each patient, the two regions were manually delineated by one experienced physician. The similarity between the two auto-segmentation methods was quantitatively evaluated by Dice similarity coefficient, the 95th Hausdorff distance, volume overlap error and relative volume difference, respectively. Statistical analyses were performed using the ranked Wilcoxon test. RESULTS: The average Dice similarity coefficient (±standard deviation) given by the deep-learning- based and Atlas-based auto-segmentation were 0.84 (±0.03) and 0.74 (±0.04) for CTV2, 0.79 (±0.02) and 0.68 (±0.03) for CTVnd, respectively. For the 95th Hausdorff distance, the corresponding values were 6.30±3.55 mm and 9.34±3.39 mm for CTV2, 7.09±2.27 mm and 14.33±3.98 mm for CTVnd. Besides, volume overlap error and relative volume difference could also predict the same situations. Statistical analyses showed significant difference between the two auto-segmentation methods (p<0.01). CONCLUSION: Compared with the Atlas-based segmentation approach, the deep-learning-based segmentation method performed better both in accuracy and stability for meaningful anatomical areas other than organs at risk.

[Feasibility of Evaluating Result of Auto-segmentation of Target Volumes in Radiotherapy with Medical Consideration Index].

OBJECTIVE: To explore the feasibility of using the bidirectional local distance based medical similarity index (MSI) to evaluate automatic segmentation on medical images. METHODS: Taking the intermediate risk clinical target volume for nasopharyngeal carcinoma manually segmented by an experience radiation oncologist as region of interest, using Atlas-based and deep-learning-based methods to obtain automatic segmentation respectively, and calculated multiple MSI and Dice similarity coefficient (DSC) between manual segmentation and automatic segmentation. Then the difference between MSI and DSC was comparatively analyzed. RESULTS: DSC values for Atlas-based and deep-learning-based automatic segmentation were 0.73 and 0.84 respectively. MSI values for them varied between 0.29~0.78 and 0.44~0.91 under different inside-outside-level. CONCLUSIONS: It is feasible to use MSI to evaluate the results of automatic segmentation. By setting the penalty coefficient, it can reflect phenomena such as under-delineation and over-delineation, and improve the sensitivity of medical image contour similarity evaluation.

SARS-CoV-2 promotes RIPK1 activation to facilitate viral propagation.

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is the ongoing global pandemic that poses substantial challenges to public health worldwide. A subset of COVID-19 patients experience systemic inflammatory response, known as cytokine storm, which may lead to death. Receptor-interacting serine/threonine-protein kinase 1 (RIPK1) is an important mediator of inflammation and cell death. Here, we examined the interaction of RIPK1-mediated innate immunity with SARS-CoV-2 infection. We found evidence of RIPK1 activation in human COVID-19 lung pathological samples, and cultured human lung organoids and ACE2 transgenic mice infected by SARS-CoV-2. Inhibition of RIPK1 using multiple small-molecule inhibitors reduced the viral load of SARS-CoV-2 in human lung organoids. Furthermore, therapeutic dosing of the RIPK1 inhibitor Nec-1s reduced mortality and lung viral load, and blocked the CNS manifestation of SARS-CoV-2 in ACE2 transgenic mice. Mechanistically, we found that the RNA-dependent RNA polymerase of SARS-CoV-2, NSP12, a highly conserved central component of coronaviral replication and transcription machinery, promoted the activation of RIPK1. Furthermore, NSP12 323L variant, encoded by the SARS-CoV-2 C14408T variant first detected in Lombardy, Italy, that carries a Pro323Leu amino acid substitution in NSP12, showed increased ability to activate RIPK1. Inhibition of RIPK1 downregulated the transcriptional induction of proinflammatory cytokines and host factors including ACE2 and EGFR that promote viral entry into cells. Our results suggest that SARS-CoV-2 may have an unexpected and unusual ability to hijack the RIPK1-mediated host defense response to promote its own propagation and that inhibition of RIPK1 may provide a therapeutic option for the treatment of COVID-19.

Dysregulated hematopoiesis in bone marrow marks severe COVID-19.

Severe coronavirus disease 2019 (COVID-19) is often indicated by lymphopenia and increased myelopoiesis; however, the underlying mechanism is still unclear, especially the alteration of hematopoiesis. It is important to explore to what extent and how hematopoietic stem cells contribute to the impairment of peripheral lymphoid and myeloid compartments in COVID-19 patients. In this study, we used single-cell RNA sequencing to assess bone marrow mononuclear cells from COVID-19 patients with peripheral blood mononuclear cells as control. The results showed that the hematopoietic stem cells in these patients were mainly in the G1 phase and prone to apoptosis, with immune activation and anti-viral responses. Importantly, a significant accumulation of immature myeloid progenitors and a dramatic reduction of lymphoid progenitors in severe cases were identified, along with the up-regulation of transcription factors (such as SPI1, LMO4, ETS2, FLI1, and GATA2) that are important for the hematopoietic stem cell or multipotent progenitor to differentiate into downstream progenitors. Our results indicate a dysregulated hematopoiesis in patients with severe COVID-19.

Stilbene dimer xylosides and flavanols from the roots of Lysidice rhodostegia and their antioxidant activities.

Eight new stilbene dimer xylosides (1-8) and one new flavanol (9), along with seven known ones (10-16) were isolated from the roots of Lysidice rhodostegia. Their structures were elucidated by extensive analysis of spectroscopic data (IR, UV, HR-ESI-MS, 1D and 2D NMR), ECD calculations and acid hydrolysis. Compounds 1-16 were evaluated for their antioxidant activities using DPPH radical-scavenging assay. Especially, compounds 9 and 10 exhibited stronger antioxidant effects than the positive control (vitamin E), with IC50 values of 9.57 ± 1.30 and 13.60 ± 1.47 µM, respectively.

A virus-like particle of the hepatitis B virus preS antigen elicits robust neutralizing antibodies and T cell responses in mice.

The preS antigen of hepatitis B virus (HBV) corresponds to the N-terminal polypeptide in the large (L) antigen in addition to the small (S) antigen. The virus-like particle (VLP) of the S antigen is widely used as a vaccine to protect the population from HBV infection. The presence of the S antigen and its antibodies in patient blood has been used as markers to monitor hepatitis B. However, there is very limited knowledge about the preS antigen. We generated a preS VLP that is formed by a chimeric protein between preS and hemagglutinin (HA), and the matrix protein M1 of influenza virus. The HBV preS antigen is displayed on the surface of preS VLP. Asn112 and Ser98 of preS in VLP were found to be glycosylated and O-glycosylation of Ser98 has not been reported previously. The preS VLP shows a significantly higher immunogenicity than recombinant preS, eliciting robust anti-preS neutralizing antibodies. In addition, preS VLP is also capable of stimulating preS-specific CD8+ and CD4+ T cell responses in Balb/c mice and HBV transgenic mice. Furthermore, preS VLP immunization provided protection against hydrodynamic transfection of HBV DNA in mice. The data clearly suggest that this novel preS VLP could elicit robust immune responses to the HBV antigen, and can be potentially developed into prophylactic and therapeutic vaccines.

Five new koumine-type alkaloids from the roots of Gelsemium elegans.

Five new koumine-type alkaloids (1-5) along with six known ones were isolated from the roots of Gelsemium elegans. Their structures with absolute configurations were elucidated on the basis of NMR spectroscopy and electronic circular dichroism spectral analyses. The inhibitory effects of compounds 1-11 on the viability of three tumor cell lines (A-649, HepG2, and HuH7) were evaluated by the MTT assay.

[Nursing care of prostate cancer patients against radiative proctitisinduced by CyberKnife treatment].

OBJECTIVE: To investigate the nursing care of prostate cancer (PCa) patients againstradioactive proctitisinduced byCyberKnifetreatment. METHODS: Sixty-eightPCapatients undergoingCyberKnife treatment in the observation group receivedspecialnursing care againstradioactive proctitis. The nursing measures includedthoserelevant toCyberKnife treatment, prevention ofradioactive proctitis, skin care, and discharge guidance. Meanwhile, another 54 prostate cancer patients received traditional nursing care as controls. We compared the incidence rate and severity of radioactive proctitis between the two groups of patients. RESULTS: The incidence rate of radioactive proctitiswas markedly lower in the observation group than in the control (2.9% vs 13.0%, P<0.05), but no statistically significant difference was observed in the severity of radioactive proctitis between the two groups of patients. CONCLUSIONS: The special nursing care againstCyberKnife-induced radioactiveproctitiscan significantlyreduce the incidence of radioactive proctitis andimprove the effect of CyberKnife treatment of prostate cancer, which therefore deserves wide clinical application.

Geleganidines A-C, Unusual Monoterpenoid Indole Alkaloids from Gelsemium elegans.

The first rotameric monoterpenoid indole alkaloids (MIAs), 1a and 1b, and two unusual dimeric MIAs, 2 and 3, with new dimerization patterns, together with their putative biosynthetic intermediates 4-7, were isolated from the roots of Gelsemium elegans. Compounds 2 and 3 represent the first natural aromatic azo- and the first urea-linked dimeric MIAs, respectively. Their structures and absolute configurations were elucidated by means of NMR spectroscopy, single-crystal X-ray diffraction, and electronic circular dichroism data analyses. The interconverting mechanism of rotamers 1a and 1b was studied by density functional theory computation. Compounds 2 and 3 showed moderate cytotoxic activity against MCF-7 and PC-12 cells, respectively. In addition, a plausible biosynthesis pathway for the new alkaloids was proposed on the basis of the coexistence of their biosynthetic precursors.