Bombyx mori serpin 3 is involved in innate immunity by interacting with serine protease 7 to regulate prophenoloxidase activation.

A subfamily of conserved proteins called serpins plays crucial roles in various physiological functions, particularly in the activation pathway of the serine protease cascade, an essential component of insect innate immunity. Here, we found Bombyx mori serpin 3 (BmSerpin3) was most highly expressed in the fat body, and was up-regulated after exposure to bacteria, fungus and virus. Further, the expression of BmSerpin3 in the hemocytes, fat body, midgut of silkworm larvae, and BmN cells was up-regulated upon Bombyx mori nucleopolyhedrovirus (BmNPV) infection. Through Bac-to-Bac expression system, we obtained the active protein of BmSerpin3, and the enzyme activity assay showed that BmSerpin3 significantly inhibited the activity of both subtilisin and trypsin. In addition, BmSerpin3 could inhibit the activation of prophenoloxidase (PPO) in larvae. The knockdown of BmSerpin3 showed increased phenoloxidase (PO) activity compared to control after BmNPV infection. Ultimately, we confirmed that BmSerpin3 interacts with B. mori Serine Protease 7 (BmSP7). Hence, we hypothesize that BmSerpin3 is involved in innate immunity by interacting with BmSP7 to regulate the PPO activation cascade. Taken together, these results showed that BmSerpin3 play a role in silkworm innate immunity and lay a foundation for studying its functions.

Optimizing the dosing regimen of roxadustat in kidney transplant recipients with early post-transplant anemia.

INTRODUCTION: Roxadustat, an oral inhibitor of hypoxia-inducible factor prolyl hydroxylase domain enzymes, has been approved for the treatment of renal anemia. However, there is a lack of study on its pharmacokinetics in kidney transplant recipients (KTRs) with early posttransplant anemia (PTA). Therefore, the aim of this study is to elucidate the pharmacokinetic characteristics of roxadustat in KTRs with early PTA and optimize the dosing regimen. METHODS: A population pharmacokinetic (PopPK) analysis was performed based on 72-hour full concentration-time profiles collected from 52 Chinese KTRs. Covariates influencing exposure were assessed using stepwise covariate modelling. Monte Carlo simulations were conducted to recommend the dosing regimen for patients with different levels of covariates. RESULTS: PopPK analysis showed that the concentration-time data can be fully described by a two-compartment model. Body weight (BW) and direct bilirubin (DBIL) levels significant affected the apparent clearance of roxadustat. Based on the established model and the estimated exposures of roxadustat by Monte Carlo simulations, a recommended dosing regimen for KTRs with early PTA at varying BW and DBIL levels were developed. Roxadustat at 100 mg three times weekly were suitable for the majority of KTRs with a DBIL level around 3 µmol/L and BW between 50 and 75 kg. The required dose may need to be increased with higher BW and lower DBIL levels, while decreased with lower BW and higher DBIL levels. CONCLUSIONS: It was the first PopPK analysis of roxadustat in KTRs with early PTA, which provide a research basis for optimizing the dosing regimen.

Bmo-miR-6498-5p suppresses Bombyx mori nucleopolyhedrovirus infection by down-regulating BmPLPP2 to modulate pyridoxal phosphate content in B. mori.

The RNA interference pathway mediated by microRNAs (miRNAs) is one of the methods to defend against viruses in insects. Recent studies showed that miRNAs participate in viral infection by binding to target genes to regulate their expression. Here, we found that the Bombyx mori miRNA, miR-6498-5p was down-regulated, whereas its predicted target gene pyridoxal phosphate phosphatase PHOSPHO2 (BmPLPP2) was up-regulated upon Bombyx mori nucleopolyhedrovirus (BmNPV) infection. Both in vivo and in vitro experiments showed that miR-6498-5p targets BmPLPP2 and suppresses its expression. Furthermore, we found miR-6498-5p inhibits BmNPV genomic DNA (gDNA) replication, whereas BmPLPP2 promotes BmNPV gDNA replication. As a pyridoxal phosphate (PLP) phosphatase (PLPP), the overexpression of BmPLPP2 results in a reduction of PLP content, whereas the knockdown of BmPLPP2 leads to an increase in PLP content. In addition, exogenous PLP suppresses the replication of BmNPV gDNA; in contrast, the PLP inhibitor 4-deoxypyridoxine facilitates BmNPV gDNA replication. Taken together, we concluded that miR-6498-5p has a potential anti-BmNPV role by down-regulating BmPLPP2 to modulate PLP content, but BmNPV induces miR-6498-5p down-regulation to promote its proliferation. Our findings provide valuable insights into the role of host miRNA in B. mori-BmNPV interaction. Furthermore, the identification of the antiviral molecule PLP offers a novel perspective on strategies for preventing and managing viral infection in sericulture.

Bmo-miR-3351 modulates glutathione content and inhibits BmNPV proliferation by targeting BmGSTe6 in Bombyx mori.

MicroRNAs (miRNAs) are small non-coding RNAs that play pivotal roles in the host response to invading pathogens. Among these pathogens, Bombyx mori nucleopolyhedrovirus (BmNPV) is one of the main causes of substantial economic losses in sericulture, and there are relatively few studies on the specific functions of miRNAs in the B. mori-BmNPV interaction. Therefore, we conducted transcriptome sequencing to identify differentially expressed (DE) messenger RNAs (mRNAs) and miRNAs in the midgut of 2 B. mori strains (BmNPV-susceptible strain P50 and BmNPV-resistant strain A35) after BmNPV infection. Through correlation analysis of the miRNA and mRNA data, we identified a comprehensive set of 21 miRNAs and 37 predicted target mRNAs. Notably, miR-3351, which has high expression in A35, exhibited remarkable efficacy in suppressing BmNPV proliferation. Additionally, we confirmed that miR-3351 binds to the 3' untranslated region (3' UTR) of B. mori glutathione S-transferase epsilon 6 (BmGSTe6), resulting in its downregulation. Conversely, BmGSTe6 displayed an opposite expression pattern to miR-3351, effectively promoting BmNPV proliferation. Notably, BmGSTe6 levels were positively correlated with glutathione S-transferase activity, consequently influencing intracellular glutathione content in the infected samples. Furthermore, our investigation revealed the protective role of glutathione against BmNPV infection in BmN cells. In summary, miR-3351 modulates glutathione content by downregulating BmGSTe6 to inhibit BmNPV proliferation in B. mori. Our findings enriched the research on the role of B. mori miRNAs in the defense against BmNPV infection, and suggests that the antiviral molecule, glutathione, offers a novel perspective on preventing viral infection in sericulture.

R&D Investments, Debt Capital, and Ownership Concentration: A Three-Way Interaction and Lag Effects on Firm Performance in China's Pharmaceutical Industry.

The existing literature has yet to provide consistent evidence on the relationship between R&D investments and firm performance. The current study attempted to fill this gap in the literature by examining the effect of lag structure and the moderating role of financial governance, in terms of debt capital and ownership concentration, on the returns of R&D. Analyzing a sample of China's pharmaceutical firms from 2009 to 2018, we found that the effect of R&D upon growth begins in the second year after R&D spending and increases thereafter. There exists a vigorous debate about the choice between debt and ownership structure. To fill this gap, we proposed a three-way interactive effect. The results suggest that firms that invest heavily in R&D may achieve their highest performance when the use of debt capital and the extent of ownership concentration are both low. This study contributes to the R&D investments and financial governance literature by reconciling previous mixed evidence about the returns of R&D and the debt-equity choices on R&D investment decisions.

Low-Voltage Actuator with Bilayer Structure for Various Biomimetic Locomotions.

Composites based on a shape-memory polymer doped with conductive particles are considered as soft actuators for artificial muscles and robots. Low-voltage actuating is expected to reduce equipment requirement and safety hazards, which requires a highly conductive particle content but weakens the reversible deformation. The spatial distribution of the conductive particle is key to decreasing the actuating voltage and maintaining the reversible deformation. Herein, an approach of fabricating a low-voltage actuator that can perform various biomimetic locomotions by spraying and hot pressing is reported. Carbon nanotubes (CNTs) are enriched inside the surface layer of poly(ethylene-co-vinyl acetate) (EVA) to form a high-density conductive network without degradation of the reversible deformation. The bilayer CNT/EVA actuator exhibits a reversible transformation of more than 10% even with 100 cycles, which requires an applied voltage of just 15 V. Taking advantage of the reprogrammability of the CNT/EVA actuator and reversible shift between the different shapes, different biomimetic locomotions (sample actuator, gripper, and walking robot) are demonstrated without any additional mechanical components. A scheme combining the electrical properties and the shape-memory effect provides a versatile strategy to fabricate low-voltage-actuated polymeric actuators, providing inspiration in the development of electrical soft actuators and biomimetic devices.

A Healable and Mechanically Enhanced Composite with Segregated Conductive Network Structure for High-Efficient Electromagnetic Interference Shielding.

HIGHLIGHTS: The cationic waterborne polyurethanes microspheres with Diels-Alder bonds were synthesized for the first time. The electrostatic attraction not only endows the composite with segregated structure to gain high electromagnetic-interference shielding effectiveness, but also greatly enhances mechanical properties. Efficient healing property was realized under heating environment. It is still challenging for conductive polymer composite-based electromagnetic interference (EMI) shielding materials to achieve long-term stability while maintaining high EMI shielding effectiveness (EMI SE), especially undergoing external mechanical stimuli, such as scratches or large deformations. Herein, an electrostatic assembly strategy is adopted to design a healable and segregated carbon nanotube (CNT)/graphene oxide (GO)/polyurethane (PU) composite with excellent and reliable EMI SE, even bearing complex mechanical condition. The negatively charged CNT/GO hybrid is facilely adsorbed on the surface of positively charged PU microsphere to motivate formation of segregated conductive networks in CNT/GO/PU composite, establishing a high EMI SE of 52.7 dB at only 10 wt% CNT/GO loading. The Diels-Alder bonds in PU microsphere endow the CNT/GO/PU composite suffering three cutting/healing cycles with EMI SE retention up to 90%. Additionally, the electrostatic attraction between CNT/GO hybrid and PU microsphere helps to strong interfacial bonding in the composite, resulting in high tensile strength of 43.1 MPa and elongation at break of 626%. The healing efficiency of elongation at break achieves 95% when the composite endured three cutting/healing cycles. This work demonstrates a novel strategy for developing segregated EMI shielding composite with healable features and excellent mechanical performance and shows great potential in the durable and high precision electrical instruments.

Phase I clinical study of personalized peptide vaccination combined with radiotherapy for advanced hepatocellular carcinoma.

AIM: To assess the efficacy and safety of a new treatment modality, cellular immune therapy based on personalized peptide vaccination (PPV-DC-CTL) combined with radiotherapy, for treating advanced hepatocellular carcinoma (HCC). METHODS: A total of nine patients with advanced HCC were enrolled. Multidisciplinary consultation confirmed that all the patients definitely had no opportunity of surgery, because four patients had multiple liver metastases (the number of liver lesions > 3), one patient had liver metastases and portal vein tumor thrombosis, one patient had lung and bone metastases, two patients had liver and lung metastases and one patient had liver metastasis and peritoneal metastasis. Patients with metastasis were treated with precise radiotherapy combined with PPV-DC-CTL. RESULTS: Following radiotherapy and one to three cycles of PPV-DC-CTL treatment, AFP levels were significantly decreased in six patients and imaging assessment of the lesions showed a partial response (PR) in three patients and stable disease in the other three patients. The response rate was 33% and disease control rate was 66%. This regimen was found to be safe and well tolerated. None of the patients developed liver or kidney side effects. Only one patient developed grade II bone marrow suppression and the remaining patients had no significant hematological side effects. CONCLUSION: Radiotherapy combined with PPV-DC-CTL provides a new therapeutic strategy for patients with advanced HCC, which is well tolerated, safe, feasible and effective.

Upregulation of Ih expressed in IB4-negative Aδ nociceptive DRG neurons contributes to mechanical hypersensitivity associated with cervical radiculopathic pain.

Cervical radiculopathy represents aberrant mechanical hypersensitivity. Primary sensory neuron's ability to sense mechanical force forms mechanotransduction. However, whether this property undergoes activity-dependent plastic changes and underlies mechanical hypersensitivity associated with cervical radiculopathic pain (CRP) is not clear. Here we show a new CRP model producing stable mechanical compression of dorsal root ganglion (DRG), which induces dramatic behavioral mechanical hypersensitivity. Amongst nociceptive DRG neurons, a mechanically sensitive neuron, isolectin B4 negative Aδ-type (IB4(-) Aδ) DRG neuron displays spontaneous activity with hyperexcitability after chronic compression of cervical DRGs. Focal mechanical stimulation on somata of IB4(-) Aδ neuron induces abnormal hypersensitivity. Upregulated HCN1 and HCN3 channels and increased Ih current on this subset of primary nociceptors underlies the spontaneous activity together with neuronal mechanical hypersensitivity, which further contributes to the behavioral mechanical hypersensitivity associated with CRP. This study sheds new light on the functional plasticity of a specific subset of nociceptive DRG neurons to mechanical stimulation and reveals a novel mechanism that could underlie the mechanical hypersensitivity associated with cervical radiculopathy.

Expression of the NLRP3 Inflammasome in Carotid Atherosclerosis.

BACKGROUND: The aim of the present study was to investigate NLRP3 inflammasome expression in human carotid atherosclerotic plaques and its relationship to plaque vulnerability. METHODS: Carotid atherosclerotic plaques collected from 30 patients scheduled for carotid endarterectomy (CEA) were subjected to immunohistochemical, mRNA, and protein expression studies. Ten mesenteric arteries from intestinal cancer patients served as controls for the immunohistochemical studies. Twenty individuals who had no carotid stenosis or coronary artery stenosis served as controls for analyzing atherosclerotic risk factors and for enzyme-linked immunosorbent assay (ELISA) studies. Serum samples were collected from all patients to determine interleukin-1ß (IL-1ß) and IL-18 levels. RESULTS: The NLRP3 inflammasome signaling pathway components NLRP3, ASC, caspase-1, IL-1ß, and IL-18 were strongly expressed in carotid atherosclerotic plaques, but not in healthy mesenteric arteries. Immunohistochemical, mRNA, and protein expression studies revealed higher expression levels of NLRP3, ASC, caspase-1, IL-1ß, and IL-18 in unstable compared to stable plaques. The NLRP3 inflammasome was localized in the cytoplasm of macrophages and foam cells and was associated with cholesterol crystal clefts inside and outside of cells. ELISA showed that the serum levels of the cytokines IL-1ß and IL-18 were higher in the CEA group compared to controls. CONCLUSIONS: These results demonstrated for the first time the close relationship between the expression of NLRP3 signaling pathway and human carotid atherosclerotic plaques. NLRP3, ASC, caspase-1, IL-1ß, and IL-18 were associated with plaque vulnerability and atherogenesis. The serum levels of IL-1ß and IL-18 may be useful predictors of atherosclerosis.

A multiple cavity malignancy involving the renal capsule, pleura and meninges: A case report and review of the literature.

Malignant renal subcapsular effusions commonly arise from primary or metastatic renal neoplasms. The current case report presents a rare case of malignancy with a massive renal subcapsular effusion accompanied by a malignant pleural effusion of an unknown primary site, which underwent progression to carcinomatous meningitis during chemotherapy. The type of adenocarcinoma present was determined by effusion cytology. Intravenous chemotherapy (docetaxel plus oxaliplatin and gemcitabine plus cisplatin) were administered; however, the disease still progressed. Time to progression was 9 months during treatment of gefitinib. Comprehensive therapies, including intracavity chemotherapy, immunotherapy and gefitinib, were shown to be effective and prolonged the patient's survival time.

Nitric oxide is involved in the regulation of trehalose accumulation under heat stress in Pleurotus eryngii var. tuoliensis.

Little is known about the mechanism of how trehalose responds to various abiotic stresses although trehalose is considered as an important protectant in fungi. We investigated the role of nitric oxide (NO) in regulating trehalose accumulation during heat stress in Pleurotus eryngii var. tuoliensis. The addition of 100 or 200 g trehalose/l significantly inhibited the production of thiobarbituric acid-reactive substance under heat stress in mycelial cells. High temperature induced endogenous trehalose accumulation and sodium nitroprusside, a NO donor, further enhanced trehalose accumulation. Finally, heat-induced trehalose accumulation could be arrested by the NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1-1-oxyl-3-oxide, at 250 µM by inhibiting the transcription of trehalose phosphate synthase gene. Thus NO plays an important role in the regulation of trehalose accumulation during abiotic stresses in P. eryngii var. tuoliensis.

Bilateral mechanical and thermal hyperalgesia and tactile allodynia after chronic compression of dorsal root ganglion in mice.

OBJECTIVE Low back pain is one of the most inextricable problems encountered in clinics. Animal models that imitate symptoms in humans are valuable tools for investigating low back pain mechanisms and the possible therapeutic applications. With the development of genetic technology in pain field, the possibility of mutating specific genes in mice has provided a potent tool for investigating the specific mechanisms of pain. The aim of the present study was to develop a mouse model of chronic compression of dorsal root ganglion (CCD), in which gene mutation can be applied to facilitate the studies of chronic pain. METHODS Chronic compression of L4 and L5 dorsal root ganglia was conducted in mice by inserting fine stainless steel rods into the intervertebral foramina, one at L4 and the other at L5. Mechanical allodynia and thermal hyperalgesia were examined with von Frey filaments and radiating heat stimulator, respectively. RESULTS The CCD mice displayed dramatic mechanical and thermal hyperalgesia as well as tactile allodynia in the hindpaw ipsilateral to CCD. In addition, this mechanical and thermal hyperalgesia as well as tactile allodynia was also found to spread to the contralateral hindpaw. CONCLUSION This model, combined with the possible genetic modification, will strengthen our knowledge of the underlying mechanisms of low back pain. It also favors the development of new treatment strategies for pain and hyperalgesia after spinal injury and other disorders which affect the dorsal root ganglion in humans.

Carbon monoxide improves adaptation of Arabidopsis to iron deficiency.

Carbon monoxide (CO) is an endogenous gaseous molecule and regulates a variety of biological processes in animals. However, whether CO regulates nutrient stress responses in plants is largely unknown. In this paper, we described an observation that CO can regulate iron-homeostasis in iron-starved Arabidopsis. Exogenous CO at 50 microm was able to prevent the iron deficient-induced chlorosis and improve chlorophyll accumulation. Expression of AtIRT1, AtFRO2, AtFIT1 and AtFER1 was up-regulated by CO exposure in iron-deficient seedlings. CO-regulated iron homeostasis could also be found in monocot maize and green alga Chlamydomonas reinhardtii. Treatment with external CO increased iron accumulation in iron-deficient Arabidopsis and C. reinhardtii, and restored leaf greening in Maize ys1 and ys3 mutants (defective in Fe uptake). Moreover, endogenous CO level was increased in Arabidopsis under iron-deficiency. Finally, CO exposure induced NO accumulation in root tips. However, such an action could be blocked by NO scavenger cPTIO. These results indicate that CO may play an important role in improving plant adaptation to iron deficiency or cross-talking with NO under the iron deficiency.

Identification of iron-deficiency responsive microRNA genes and cis-elements in Arabidopsis.

MicroRNAs (miRNAs) are a class of endogenous non-coding small RNAs that bind to their target mRNAs to repress their translation or induce their degradation. Recent studies have shown that several miRNAs regulate plant adaptation to sulfate and phosphate deficiency. However, whether miRNAs are involved in regulation of stress response to iron (Fe) deficiency is unknown. In this study, we carried out a survey of Arabidopsis miRNA genes in response to Fe deficiency and identified IDE1/IDE2 (Iron-deficiency responsive cis-Element 1 and 2) in their promoter regions. We constructed a small RNA library from Arabidopsis seedlings under Fe deficiency. Sequence analysis revealed 8 conserved miRNA genes in 5 families, all of which were up-regulated during Fe deficiency. Further, we analyzed cis-regulatory elements upstream of all miRNA genes in Arabidopsis and found 24 miRNA genes containing IDE1/IDE2 motifs in their promoter regions. Transcriptional analysis using RT-PCR showed that 70.8% (17/24) of the IDE-containing miRNA genes were expressed in response to Fe deficiency. We presented a putative interaction model between protein-coding genes and miRNA genes under Fe deficiency. Our analytic approach is useful and efficient because it is applicable to cis-element finding for miRNAs responding to other abiotic stresses. Also, the data obtained in this study may aid our understanding of the role of Fe deficiency responsive specific sequences upstream of miRNA genes and the functional implications of miRNA genes in response to Fe stress in plants.

Study on the efficacy and safety of Xueyou Mixture in treating hemophilia.

OBJECTIVE: To observe the effect of Xueyou Mixture (, XYM) on blood coagulation factors and its safety in treating hemophilia. METHODS: To the randomly selected 65 inpatients of hemophilia, XYM was administered accompanied with intravenous dripping of liver cell growth factor 60-100 mg once a day to protect the liver, with no blood products like concentrated VIII and FIX factors or blood plasma given. The treatment lasted for 3 weeks. The short-term efficacy and adverse reactions were observed. The long-term efficacy in patients was observed in a follow-up study of 6-12 months after they were discharged from the hospital but continuously took XYM orally. RESULTS: The short-term markedly effective rate in the patients was 95.38% (62/65). After they were treated for 3 weeks, the level of FVIII factor activity increased in 56 patients of type A from (3.32+/-2.21) % to (4.18+/-2.23) %, and in 9 of type B from (4.92+/-1.81) % to (5.64+/-1.96) %. Compared with that before treatment, the difference was significant in both of them (P<0.01). No obvious adverse reaction was found in the treatment period. The follow-up study showed that in 22 patients of type A, the FVIII factor activity ratio increased from (3.25+/-2.11) % to (6.31+/-2.16) %, (8.36+/-1.05) %, and (16.38+/-2.71) % in the 2nd, 3rd and 6th month after discharge respectively, all showing significant difference to that before treatment (P<0.01); and in 4 patients of type B, it increased from (4.15+/-2.26) % to 7.8% and 11.6% (mean value) in the 2nd and 6th month respectively. CONCLUSION: XYM could raise the activity of factors VIII and IX in patients with hemophilia, and the degree of the rise is related with the duration of the therapy, with no obvious adverse reaction, which strikes out a new path and new train of thinking for the treatment of the disease by nonblood preparation.

Study on graded therapy of hemophilic arthritis by integrative traditional Chinese and Western medicine.

OBJECTIVE: To study the effect and safety of graded therapy featuring integrative traditional Chinese and Western medicine for the treatment of hemophilic arthritis. METHODS: Forty patients with hemophilic arthritis were hospitalized randomly, with their blood coagulation factor activity determined by one-stage method and their arthritis classified into 4 stages. The treatment was applied according to the stage of arthritis and finding of intra-articular cavity puncture. For stage I, based on the principle of RICE (rest, ice, compression and elevation), 1.8 g of Xuefuda was medicated orally once per day, intravenous dripping of 250 mL of hemostasis mixture twice a day and 1.2 g of clindamycin per day were also given for hemostasis and anti-inflammation. For stage II-III, Kangyanling was additionally administered via intra-articular cavity injection twice a week, 2 mL every time, for 5-6 times in total. For stage IV, the drug for intra-articular cavity injection was replaced with 25 mg of sodium hyaluronate and the frequency of injection reduced to every two weeks, for 5-6 times in total. Coagulation factors III and IV as well as blood plasma were not given in the whole treatment course. Short-term therapeutic effects and adverse reaction in patients were evaluated, and the long-term effects were followed-up after patients left the hospital with 6-month consolidation therapy by Xuefuda. RESULTS: After a 3-week treatment, 33 patients (82.5%) were completely remitted; 5 (12.5%) were partially remitted and 2 (5.0%) un-remitted, setting the short-term effective rate at 95.0% (38 cases). The 6-month follow-up showed that except for a relapse in 2 and 4 patients of stage III and IV respectively, long-term remission displayed in all the other 34 patients, with the remission sustaining rate being 85.0%. No complication such as an infection, bleeding or aggravating pain occurred in the 215 times intra-articular puncturing conducted in the 40 patients. Normal figures were shown in liver and kidney function, electrolytes, ECG, blood glucose and routine test of blood and urine throughout the course. CONCLUSION: The graded treatment of integrative medicine for hemophilia with non-blood preparation has a favorable effect and is safe or without any adverse reaction, which opens a high efficacy and new safe path and thinking for the treatment of and deformity prevention in the hemophilic patients.