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In recent years, it has been seen that artificial intelligence (AI) starts to bring revolutionary changes to chemical synthesis. However, the lack of suitable ways of representing chemical reactions and the scarceness of reaction data has limited the wider application of AI to reaction prediction. Here, we introduce a novel reaction representation, GraphRXN, for reaction prediction. It utilizes a universal graph-based neural network framework to encode chemical reactions by directly taking two-dimension reaction structures as inputs. The GraphRXN model was evaluated by three publically available chemical reaction datasets and gave on-par or superior results compared with other baseline models. To further evaluate the effectiveness of GraphRXN, wet-lab experiments were carried out for the purpose of generating reaction data. GraphRXN model was then built on high-throughput experimentation data and a decent accuracy (R 2 of 0.712) was obtained on our in-house data. This highlights that the GraphRXN model can be deployed in an integrated workflow which combines robotics and AI technologies for forward reaction prediction., (© 2023. Springer Nature Switzerland AG.)

Anaplastic lymphoma kinase ( ALK ) rearrangements are one of the most common mutations in nonsmall cell lung cancer. Majority of the ALK rearrangements were sensitive to crizotinib, yet some rare fusion types may less benefit. The patient with LINC01923-ALK fusion was treated with crizotinib for 1 week and developed an adverse rash reaction. Replaced with second-line treatment with esatinib, the patient had a partial response in the primary site and achieved a complete response in the brain metastases. The patient was treated successfully with ensartinib leading to a progression-free survival of 6 months (and counting). This is the first report on one lung adenocarcinoma patient with a novel LINC01923-ALK fusion beneficial from ensartinib, which provides more knowledge for ALK fusion spectrum., (Copyright © 2022 Wolters Kluwer Health, Inc. All rights reserved.)

Shrimp are especially susceptible to the White Spot Syndrome Virus (WSSV). Oral administration of the WSSV envelop protein VP28 is a promising approach to protect shrimp against WSSV. In this study, Macrobrachium nipponense (M. nipponense) were fed for 7 days with food supplemented with Anabaena sp. PCC 7120 (Ana7120) expressing VP28 and then challenged with WSSV. The survival rates of M. nipponense in three groups, including control, WSSV-challenged, and VP28-vaccinated, were subsequently determined. We also determined the WSSV content of different tissues and the tissue morphology in the absence of and after viral challenge. The survival rate of the positive control group (no vaccination and challenge, 10%) and empty vector group (fed with Ana7120 pRL-489 algae and challenged, 13.3%) was much lower than the survival rate of M. nipponense in wild type group (fed with Ana7120 and challenged, 18.9%), immunity group 1 (fed with 3.33% Ana7120 pRL-489-vp28 and challenged, 45.6%) or immunity group 2 (fed with 6.66% Ana7120 pRL-489-vp28 and challenged, 62.2%). RT-qPCR showed that WSSV content of the gill, hepatopancreas and muscle of immunity groups 1 and 2 were substantially lower than the positive control. Microscopic examination revealed that WSSV-challenged positive control exhibited large number of cell rupture, necrosis, nuclear exfoliation in gills and hepatopancreatic tissues. The gill and hepatopancreas of immunity group 1 showed partial symptoms of infection, yet the tissue was visibly healthier than that of the positive control group. No symptoms were visible in the gills and hepatopancreatic tissue of immunity group 2. The results demonstrate that the probability of M. nipponense infected by WSSV can be diminished by oral administration of cyanobacteria-expressed VP28. Such an approach could improve the disease resistance and delay the death of M. nipponense in the commercial production of this shrimp., Competing Interests: Declaration of competing interest The authors have no competing interests to be declared., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Atopic dermatitis (AD) is the most prevalent chronic inflammatory skin condition and significantly reduces quality of life. Tight junction (TJ), which is located directly beneath the stratum corneum, maintains skin barrier function and aids in the identification of the cell's "territory". We evaluated seventeen TJ related genes to explore AD related alterations of TJ. Remarkably, we found that the expression of ZO-3, a gene that had not been linked to the development of TJ in AD, was significantly down-regulated in the skin of AD mice and patients. siRNA mediated knock-down of ZO-3 significantly decreased transepithelial electrical resistance in HaCaT cells, demonstrating that ZO-3 is essential to epidermal barrier function. In addition to ZO-3 downregulation, protein kinase B (Akt) phosphorylation was increased in the skin of AD mice. We further confirmed an inverse relationship between Akt phosphorylation and ZO-3 expression in AD using HaCaT cells and mouse model. Finally, we tested the efficacy of osthole as a treatment for AD in mice and HaCaT cells. Osthole inhibits Akt phosphorylation, and thereby enhances ZO-3 expression in mouse models of AD, resulting in greatly lessened AD associated skin damage and chronic itch, and osthole also increased the expression of ZO-3 in HaCaT cells by inhibiting the phosphorylation of Akt. Together, we established that ZO-3 is essential for the development of TJ in AD skin and HaCaT cells, and our findings provide fresh support for osthole's ability to protect ZO-3 expression and the epidermal barrier in AD., Competing Interests: Declaration of competing interest None., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Background: Curcumin is a diketone compound extracted from the rhizomes of some plants in the Zingiberaceae and Araceae family. It possesses a variety of biological activities, including antioxidant, anti-inflammatory and anti-cancer properties. However, the cellular and molecular antipruritic mechanisms of curcumin remain to be explored., Objective: Our objective was to study the role of curcumin in pruritus and determine whether its antipruritic effect is related to MrgprB2 receptor., Methods: The effect of curcumin on pruritus in mice was examined by scratching behavior test. The antipruritic mechanism of curcumin was explored by using transgenic mice (MrgprB2 -/- mice, MrgprB2Cre Td/tomato mice), histological analysis, western blot and immunofluorescence. In addition, the relationship between curcumin and MrgprB2/X2 receptor was studied in vitro by using calcium imaging, plasmid transfection and molecular docking RESULTS: In the current study, we found that curcumin had obvious antipruritic effect. Its antipruritic effect was related to the regulation of MrgprB2 receptor activation and mast cells tryptase release. In vitro, mouse peritoneal mast cells activated by compound 48/80 could be inhibited by curcumin. In addition, curcumin was also found to suppress the calcium flux in MrgprX2 or MrgprB2-overexpression HEK cells induced by compound 48/80, substance P, and PAMP 9-20, displaying the specific relation with the MrgprB2/X2 receptor. Moreover, molecular docking results showed that curcumin had affinity to MrgprX2 protein., Conclusions: Overall, these results indicated that curcumin has the potential to treat pruritus induced by mast cell MrgprB2 receptor., (© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Rutin is a natural anti-inflammatory ingredient widely found in medicinal plants. Studies have shown that rutin inhibits mast cell degranulation and the release of inflammatory mediators. Mast cell P2X7 receptor mediates mast cell degranulation and serves as a therapeutic target for inflammatory pain. Herein, the aim of this study was to investigate whether the anti-inflammatory mechanism of rutin is related to the mast cell P2X7 receptor. Our results showed that rutin could inhibit [Ca 2+ ] i elevation induced by 5 mM ATP or 30 μM BZATP in a concentration-dependent manner in mouse peritoneal mast cells. Rutin also suppressed the inward current mediated by P2X7 receptor. In vivo, rutin could significantly inhibit the mechanical hypersensitivity induced by 100 mM ATP that is associated with P2X7 receptor in mast cells. Moreover, molecular docking revealed the high affinity between rutin and the P2X7 receptor crystal structure. Collectively, this study demonstrated that rutin attenuated inflammatory pain by inhibiting the activity of P2X7 receptor in mast cells., (© 2022. The Author(s) under exclusive licence to University of Navarra.)

Purpose: Keloid is a type of benign fibrous proliferative tumor characterized by excessive scarring. C1q/TNF-related protein 3 (CTRP3) has been proven to possess antifibrotic effect. Here, we explored the role of CTRP3 in keloid. In the current research, we examined the influence of CTRP3 on keloid fibroblasts (KFs) and investigated the potential molecular mechanism., Methods: KF tissue specimens and adjacent normal fibroblast (NF) tissues were collected cultured from 10 keloid participants. For the TGF- β 1 stimulation group, KFs were processed with human recombinant TGF- β 1. Cell transfection of pcDNA3.1-CTRP3 or pcDNA3.1 was performed. The siRNA of CTRP3 (si-CTRP3) or negative control siRNA (si-scramble) was transfected into KFs., Results: CTRP3 was downregulated in keloid tissues and KFs. CTRP3 overexpression significantly controlled TGF- β 1-induced propagation and migration in KFs. Col I, α -SMA, and fibronectin mRNA and protein levels were enhanced by TGF- β 1 stimulation, whereas they were inhibited by CTRP3 overexpression. In contrast, CTRP3 knockdown exhibited the opposite effect. In addition, CTRP3 attenuated TGF- β receptors TRI and TRII in TGF- β 1-induced KFs. Furthermore, CTRP3 prevented TGF- β 1-stimulated nuclear translocation of smad2 and smad3 and suppressed the expression levels of p-smad2 and p-smad3 in KFs., Conclusion: CTRP3 exerted an antifibrotic role through inhibiting proliferation, migration, and ECM accumulation of KFs via regulating TGF- β 1/Smad signal path., Competing Interests: The authors declare that they have no conflict of interest., (Copyright © 2023 Lin He et al.)

Background & Aims: Capsaicin receptor, also known as transient receptor potential vanilloid 1 (TRPV1), is involved in pain physiology and neurogenic inflammation. Herein, we discovered the presence of TRPV1 in hepatic stellate cells (HSCs) and aimed to delineate its function in this cell type and liver fibrosis., Methods: TRPV1 expression was examined in liver biopsies from patients with liver fibrosis using quantitative real-time PCR and immunostaining. Its contribution to liver fibrosis was examined in Trpv1 -/- mice, upon lentiviral delivery of the TRPV1 gene, and in human and mouse primary HSCs, using patch clamp, intracellular Ca 2+ mobilization determination, FACS analyses and gain/loss of function experiments. Binding of sterile alpha and Toll/interleukin-1 receptor motif-containing protein 1 (SARM1) to TRPV1 was determined using mass spectrometry, co-immunoprecipitation, surface plasmon resonance, bioluminescence resonance energy transfer, and NanoBiT., Results: TRPV1 mRNA levels are significantly downregulated in patients with liver fibrosis and mouse models, showing a negative correlation with F stage and α-smooth muscle actin expression, a marker of HSC activation. TRPV1 expression and function decrease during HSC activation in fibrotic livers in vivo or during culture. Genetic and pharmacological inhibition of TRPV1 in quiescent HSCs leads to NF-κB activation and pro-inflammatory cytokine production. TRPV1 requires binding of its N-terminal ankyrin repeat domain to the TIR-His583 (Toll/interleukin-1 receptor) domain of SARM1 to prevent HSCs from pro-inflammatory activation. Trpv1 -/- mice display increased HSC activation and more severe liver fibrosis, whereas TRPV1 overexpression is antifibrotic in various disease models., Conclusion: The antifibrotic properties of TRPV1 are attributed to the prevention of HSC activation via the recruitment of SARM1, which could be an attractive therapeutic strategy against liver fibrosis., Impact and Implications: We identified the neuronal channel protein TRPV1 as a gatekeeper of quiescence in hepatic stellate cells, a key driver of liver fibrogenesis and chronic liver disease. Physiologically expressed in healthy liver and consistently downregulated during liver fibrosis development, its therapeutic re-expression is expected to have few side effects, making it an attractive target diagnostic tool and drug candidate for industry and clinicians., Competing Interests: Conflict of interest All authors declare that they have no conflict of interest. Please refer to the accompanying ICMJE disclosure forms for further details., (Copyright © 2023 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

Diabetic nephropathy is one of the major causes of end-stage renal disease, and the pathogenesis of the disease has not been elucidated. While the immunoinflammatory response plays an essential role in the progression of diabetic nephropathy. Glomerular expression dataset in diabetic nephropathy was obtained from the GEO database. Differentially expressed genes were identified and functional enrichment analysis was performed to find genes associated with immunity and inflammation from them. The hub genes of immunoinflammatory were identified using MCODE after establishing the PPI network and gene expression was verified with diabetic nephropathy model rats. Xcell was used to assign immune cells to diabetic nephropathy glomerular samples to detect significant changes in immune cells and to analyze correlations with the hub gene. We found 120 DEGs associated with immunity and inflammation, Ribonuclease 6 was the Hub gene with the highest MCODE score. Xcell analysis revealed significant changes of immune cells in DN glomeruli, including upregulated Activated DCs, Conventional DCs, CD4+ Tem, Epithelial cells, Macrophages, Macrophages M1, and Memory B-cells. RNase6 expression showed the highest positive correlation with Macrophages M1, Activated DCs, and Conventional DCs. We verified through the Nephroseq v5 database that RNase6 expression was elevated in DN glomeruli and negatively correlated with glomerular filtration rate. Animal studies revealed that the kidney of DN model rats showed increased RNase6 expression together with inflammatory factor TNF-alpha and chemokine MCP-1. Our study identified RNase6 as a diagnostic and prognostic biomarker for diabetic nephropathy and found that it may play an essential role in the immunoinflammatory damage to the glomerulus., (© 2022. The Author(s).)

This study aims to investigate the spatial and temporal characteristics, pollution degrees, and potential ecological risks of mercury (Hg) in urban lake waters and sediments in Guangzhou, where is a typical area of Hg emission and population-economic-industrial concentration in South China. In different districts of this city, the water from 15 lakes were collected continuously from June 2020 to May 2021, and the sediments from 9 lakes were collected in 2015 and 2021. The seasonal changes of Hg concentration (Hg -C ) in the water were found to be high in winter and low in summer. The spatial distribution of Hg -C in sediments showed that it was high in urban central areas and low in suburbs. The Nemero index and geological accumulation index showed that there were uncontaminated of Hg in the collected lake water, and above moderately contaminated in the lake sediments in urban center, respectively. The Hg pollution potential ecological risk index showed that there was low risk in the collected water, high and extremely high risk in the lake sediments in urban center, respectively. The principal component analysis (PCA) and correlation analysis (CA) of Hg and meteorological factors showed that precipitation, temperature, and vapor pressure had negative effects on the seasonal changes of Hg -C in water, and air pressure and wind direction had positive effects. The PCA and CA of Hg and other geochemical elements showed that anthropogenic emissions may be the main sources of Hg in sediments, which was also supported by the data of population density, road density, and motor vehicles per 1000 people. This study provided a reference for urban lake pollution treatment, resident health, and ecological environment protection., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

A novel mast cell-specific G-protein-coupled receptor (GPCR), known as Mas-related G protein-coupled receptor-B2 (MRGPRB2), plays important roles in immune response. However, the opening of ion channels mediated by MRGPRB2 activation remains unclear. In this study, we found that [Ca 2+ ]i elevation and voltage-dependent current generated by MRGPRB2 activation were correlated with extracellular calcium concentration. The increases in [Ca 2+ ]i and voltage-dependent current caused by MRGPRB2 activation were blocked by U73122 (PLC blocker) or 2-APB (IP3 blocker) or synta66 (ORAI blocker). The voltage-dependent current induced by MRGPRB2 was inhibited by calcium-activated chlorine channel (CACCS) blockers, DIDS, or NPPB. Our results indicated the involvement of the PLC-IP3-ORAI signaling pathway and CACCS in MRGPRB2-mediated mast cell activation., (Copyright © 2022. Published by Elsevier B.V.)

Neurofibromatosis type 1 (NF1) is a kind of common neurogenetic disorder associated with various developmental deficits. Circular RNAs (circRNAs) have been frequently verified to be crucial modulators in human diseases. However, the functions of circRNAs on the occurrence of NF1 remain largely obscure. In our study, RT-qPCR was applied to analyze circ_0061,587 expression and we noticed that circ_0061,587 expression was overtly elevated in human NF1-associated malignant peripheral nerve sheath tumor (MPNST) cell lines. Meanwhile, the results of loss-of-function assays revealed that silencing of circ_0061,587 hampered the proliferation, migration, and invasion but stimulated the apoptosis of human NF1-associated MPNST cells. In addition, mechanism assays were implemented to unveil the possible regulatory mechanism behind circ_0061,587. As a result, circ_0061,587 sequestered microRNA-485-5p (miR-485-5p) to modulate the expression of RUNX family transcription factor 1 (RUNX1) and annexin A11 (ANXA11). Finally, rescue experiments confirmed that circ_0061,587 boosted the malignant behaviors of human NF1-associated MPNST cells through up-regulating RUNX1 and ANXA11. In conclusion, circ_0061,587 functioned as an oncogene in NF1-associated MPNST cells and this study might provide novel insights for the diagnosis and treatment of NF1., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Background: Breast milk is recognized as the best source of nutrition for babies. The World Health Organization (WHO) recommends exclusive breastfeeding in the first six months after birth and continued breastfeeding for up to two years. According to the reported literature, breastfeeding and exclusive breastfeeding rates worldwide are relatively low, especially after a cesarean section (C-section). Therefore, this review aims to summarize existing data on C-section and breastfeeding performance worldwide to interpret their relationship further., Methods: Research articles related to C-section and breastfeeding were retrieved from electronic databases, including CINAHL Complete, Health Source: Nursing/Academic Edition, Academic Search Complete, MEDLINE, PubMed, and Google Scholar. Only full-text English articles reported from 2015 to 2020 are summarized in this review., Results: Among a total of 389 articles identified, 18 papers met our inclusion criteria, which reported that the C-section was associated with the initiation of breastfeeding and the duration of exclusive breastfeeding. Furthermore, these studies also discussed factors and experiences related to breastfeeding difficulties in mothers who have a C-section. Besides, several studies investigated effective initiatives that support breastfeeding in mothers who have a C-section., Conclusion: C-section is thought to be related to the initiation and duration of breastfeeding. In comparison with natural childbirth, C-section can delay the start of breastfeeding and shorten the duration of exclusive breastfeeding. Moreover, the planned C-section is considered the most critical factor affecting breastfeeding. Also, breastfeeding initiatives are highly recommended to support mothers who have a C-section. According to the literature, different regions and populations may have distinct experiences of breastfeeding. Therefore, future research is required to identify breastfeeding support for diverse populations with higher quality., Competing Interests: Declaration of Competing Interest None Declared., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Psoriasis is an immune-mediated chronic inflammatory skin disease characterized by erythema, scales, and infiltration of the skin, which causes deleterious effects on patient quality of life. TRP channel played important roles in the generation and conductance of itch signal . According to our results, psoriasis induced itch was TRPV4 dependent, and TRPV4 expression in both epidermis and DRG were up-regulated in psoriasis. Thus, TRPV4 is an attractive candidate for treating psoriasis induced itch. Cimifugin is a common compound in antipruritic Chinese medicine. In our study, GSK1016790A, a TRPV4 channel specific agonist, induced acute itch was inhibited by cimifugin in a dose-dependent manner. Furthermore, cimifugin treatment reduced the scratching behavior and reversed the TRPV4 up-regulation induced by psoriasis. In particular, cimifugin decreased GSK1016790A induced calcium response both in HaCaT cells and DRG neurons. Importantly, in TRPV4 transfected HEK293 cells, GSK101 induced calcium response was also significantly inhibited by cimifugin pretreatment. Consistent with our calcium imaging result, cimifugin pretreatment also inhibited GSK101 induced inward currents. Our study delineated a new role of TRPV4 in psoriasis and emphasized the antipruritic effect of cimifugin, which opened a new avenue to itch management in psoriasis., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Asthma is an inflammatory disorder of the airways dominated by a Th2-type pattern. Recently, an emerging interest arises whether transient receptor potential ankyrin 1 (TRPA1) plays a potential role in the adaptive immune response. In this study, the role of TRPA1 in the development and exacerbation of asthma was explored. The classic OVA-induced asthma and OVA plus PM2.5-induced exacerbated asthma model were used. The CD4+ T cells were sorted from spleen in asthmatic and exacerbated asthmatic mice. In the BALB/c mice treated with OVA, the increased phenotype of asthma was obtained, accompanied by the high expression of TRPA1 in lung tissue and levels of IL-4, IL-13, NGF, PGD 2 in BAL. In contrast, genetic deletion or pharmacological inhibition of TRPA1 alleviated the phenotype of asthma. Similarly, in wild type (WT) C57BL/6 mice treated with OVA, the high expression of TRPA1 in lung tissues was obtained, and the levels of IL-4, IL-13, NGF, PGD 2 in BAL remarkably increased when compared with those in the TRPA1 deleted mice. Furthermore, high expression of TRPA1 was detected in CD4+ T cells of OVA-treated WT C57BL/6 mice. Additional detection in the asthmatic mice exacerbated by OVA plus PM2.5 also showed high TRPA1 expression in lung tissue and CD4+ T cells. All evidence confirmed that TRPA1 is essential for the development and exacerbation of asthma. More importantly, the expression of TRPA1 in CD4+ T cells of different asthmatic mice suggested that it might be involved in neuro-immune interactions in airway inflammation of asthmatic mice., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

This study aimed to assess the effect of oral administration of Synechococcus sp. PCC 7942 harboring vp19, vp28, and vp(19 + 28)against infection by white spot syndrome virus (WSSV) on juveniles of Litopenaeus vannamei. L. vannamei was orally administrated by feeding with different mutants of Synechococcus for 10 days, and then challenged with WSSV. The cumulative mortality of vp19, vp28, vp (19 + 28) groups was lower than that of the positive control group (57.8%, 62.2%, 71.1%, respectively); vp (19 + 28) group had a better protection rate than vp19 and vp28 groups. The analysis of shrimp immunological parameters showed that, after WSSV injection, the activity of superoxide dismutase, phenol oxidase, catalase, and lysozyme in the hepatopancreas of vp19, vp28, and vp (19 + 28) groups was higher than in the positive group; at the same time, growth performances of L. vannamei of experimental groups were better than control groups. Results showed that the Synechococcus mutants harboring vp19, vp28, and vp (19 + 28) could be used both as drug and feed to also enhance the defensive ability of juvenile shrimp against WSSV infection by increasing the activity of immune related enzymes., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Microcystin-LR (MC-LR), a widespread environmental contaminant, has been shown to have potent acute testicular toxicity. However, magnitudes of toxic effects, induced by MCs, depend on route and magnitude of exposure to the toxin. In the present study, male mice were orally exposed 1, 10 or 100 μg/L MC-LR for 90 or 180 days, and pathological approach and the isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomics were employed with testes. Proteomics revealed that a number of differentially altered proteins may be involved in MC-LR-induced chronic testicular toxicity. The biological process analysis indicated the altered proteins played an important role in biological adhesion, cellular process, response to stimulus or rhythmic process. The cellular component analysis revealed that most of the proteins with altered expression associated with cell part, extracellular region, extracellular region part, membrane, membrane part, organelle or organelle part. The molecular function showed that these proteins were critical in molecular transducer activity. Integrity analyses provide first compelling evidence that MC-LR significantly cause dysfunction of blood-testis barrier (BTB) through affecting tight junctions and gap junctions. Moreover, phosphatidylinositol 3-kinase (PI3K)/AKT eventually contributed to injury result from chronic low-level MC-LR treatment. Identification of proteins in testis responsive to MC-LR provides insights into molecular mechanisms of chronic toxicity of MCs., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Neuropathic pain remains a therapeutic challenge because of its complicated mechanisms. Mas-related GPCR D (MrgprD) is specifically expressed in small-diameter, nociceptive neurons of dorsal root ganglia (DRGs) and is implicated in pain modulation. However, the underlying mechanism of MrgprD involved in neuropathic pain remains elusive. In this study, we used behavioral experiments and physiologic examination methods to investigate the role of MrgprD in chronic constriction injury (CCI)-induced neuropathic pain. We found that MrgprD is necessary for the initiation of mechanical hypersensitivity and cold allodynia, but not for heat allodynia. Moreover, we demonstrated that transient receptor potential cation channel (TRP)-A1 was the ion channel downstream of MrgprD, and the β-alanine-induced calcium signal was attributed mostly to TRP-A1 function. We further showed that PKA serves as a downstream mediator of β-alanine-activated MrgprD signaling to activate TRP-A1 in DRG neurons and in human embryonic kidney 293 cells, to coexpress MrgprD and TRP-A1 plasmids. Finally, we found that the β-alanine-induced pain behavior was increased, whereas the itching behavior was unchanged in CCI models compared with sham-injured animals. Knockout of TRPA1 also attenuated the β-alanine-induced pain behavior in CCI models. In conclusion, MrgprD is essential in cold allodynia in CCI-induced neuropathic pain through the PKA-TRP-A1 pathway. TRP-A1 facilitates MrgprD to development of neuropathic pain. Our findings reveal a novel mechanism of neuropathic pain formation and highlight MrgprD as a promising drug target for the treatment of neuropathic pain.-Wang, C., Gu, L., Ruan, Y., Geng, X., Xu, M., Yang, N., Yu, L., Jiang, Y., Zhu, C., Yang, Y., Zhou, Y., Guan, X., Luo, W., Liu, Q., Dong, X., Yu, G., Lan, L., Tang, Z. Facilitation of MrgprD by TRP-A1 promotes neuropathic pain.

Aim: To investigate the association of PNPLA3 polymorphisms with concurrent chronic hepatitis B (CHB) and nonalcoholic fatty liver disease (NAFLD)., Methods: A cohort of Han patients with biopsy-proven CHB, with or without NAFLD (CHB group, n = 51; CHB + NAFLD group, n = 57), and normal controls (normal group, n = 47) were recruited from Northern (Tianjin), Central (Shanghai), and Southern (Zhangzhou) China. Their PNPLA3 polymorphisms were genotyped by gene sequencing. The association between PNPLA3 polymorphisms and susceptibility to NAFLD, and clinical characteristics of NAFLD were evaluated on the basis of physical indices, liver function tests, glycolipid metabolism, and histopathologic scoring. The association of PNPLA3 polymorphisms and hepatitis B virus (HBV) load was determined by the serum level of HBV DNA., Results: After adjusting for age, sex, and body mass index, we found that four linked single nucleotide polymorphisms (SNPs) of PNPLA3, including the rs738409 G allele (CHB + NAFLD group vs CHB group: odds ratio [OR] = 2.77, 95% confidence interval [CI]: 1.18-6.54; P = 0.02), rs3747206 T allele (CHB + NAFLD group vs CHB group: OR = 2.77, 95%CI: 1.18-6.54; P = 0.02), rs4823173 A allele (CHB + NAFLD group vs CHB group: OR = 2.73, 95%CI: 1.16-6.44; P = 0.02), and rs2072906 G allele (CHB + NAFLD group vs CHB group: OR = 3.05, 95%CI: 1.28-7.26; P = 0.01), conferred high risk to NAFLD in CHB patients. In patients with both CHB and NAFLD, these genotypes of PNPLA3 polymorphisms were associated with increased susceptibility to nonalcoholic steatohepatitis (NASH) (NAFLD activity score ≥ 3; P = 0.01-0.03) and liver fibrosis (> 1 Metavir grading; P = 0.01-0.04). As compared to those with C/C and C/G at rs738409, C/C and C/T at rs3747206, G/G and G/A at rs4823173, and A/A and A/G at rs2072906, patients in the CHB + NAFLD group with G/G at rs738409, T/T at rs3747206, A/A at rs4823173, and G/G at rs2072906 showed significantly lower serum levels of HBV DNA (P < 0.01-0.05)., Conclusion: Four linked SNPs of PNPLA3 (rs738409, rs3747206, rs4823173, and rs2072906) are correlated with susceptibility to NAFLD, NASH, liver fibrosis, and HBV dynamics in CHB patients.

Objective: To study the effects of ultrashort wave combined with sequential pressure treatment on the functional recovery of deeply burned hands in the early stage of healed wounds in hands., Methods: Sixty-five patients with burn of unilateral hand were hospitalized from July 2012 to June 2013 in our center. Injured hands of 35 patients were treated with active movement, ultrashort wave, sequential pressure therapy, and pressure gloves, and the other 30 patients were treated with active movement and pressure gloves 10-31 days after the wounds were healed according to the will of patients. The former 35 patients were regarded as comprehensive treatment (CT) group, and the latter 30 patients were regarded as routine treatment (RT) group. Before treatment and 4 weeks after treatment, the appearance of injured hands was observed; the circumference of the proximal segment of thumb, index, and middle fingers and that of the palmar crease and wrist crease were measured to evaluate swelling of injured hand; score and grade of function of injured hands were evaluated with a Carroll Upper Extremity Functional Test. Data were processed with t test and rank sum test., Results: (1) Four weeks after treatment, appearance of 30 injured hands in group CT was improved, which was close to that of the normal hand of each patient, while contracture deformity of metacarpophalangeal joints and interphalangeal joints was observed in the other 5 injured hands. Four weeks after treatment, no obvious change in the appearance of 17 injured hands in group RT was observed compared with that before treatment, while hyperextension of metacarpophalangeal joints, flexion of interphalangeal joints, and adduction deformity of thumb were observed in the other 13 hands. (2) Four weeks after treatment, the circumferential values of the proximal segment of thumb, index, and middle fingers and the palmar crease and wrist crease of injured hands in group CT were respectively lower than those before treatment (with t values 3.26-4.24, P values below 0.01), and the circumferential values of the proximal segment of thumb and middle fingers and the wrist crease of injured hands in group RT were respectively lower than those before treatment (with t values 2.02-2.44, P values below 0.05). The difference values of circumference values of the proximal segment of thumb, index, and middle fingers and the palmar crease and wrist crease of injured hands between before treatment and 4 weeks after treatment were respectively (0.491 ± 0.022), (0.583 ± 0.089), (0.486 ± 0.021), (1.100 ± 0.076), (0.751 ± 0.053) cm in group CT, which were significantly higher than those in group RT [(0.306 ± 0.021), (0.277 ± 0.022), (0.320 ± 0.027), (0.700 ± 0.052), (0.483 ± 0.048) cm, with t values respectively 5.94, 3.11, 5.02, 4.22, 3.68, P values below 0.01]. (3) Four weeks after treatment, scores of function of injured hands in groups CT and RT were respectively higher than those before treatment (with t values respectively 14.40 and 4.00, P values below 0.001), and the grades of function of injured hands were respectively improved (with u values respectively 6.93 and 3.29, P values below 0.01). The difference value of scores of function of injured hands between before treatment and 4 weeks after treatment was (51.1 ± 2.2) points in group CT, which was significantly higher than that of group RT [(32.5 ± 4.8) points, t = 3.52, P < 0.001]., Conclusions: Ultrashort wave combined with sequential pressure and routine rehabilitation treatment of deeply burned hands in the early stage after wounds in hands are healed can obviously reduce the swelling of injured hands, which provides a favorable condition for active movements and systematic rehabilitation treatment later.

Objective: To explore the effects of group psychological counseling on the self-confidence and social adaptation of burn patients during the course of rehabilitation., Methods: Sixty-four burn patients conforming to the inclusion criteria and hospitalized from January 2012 to January 2014 in Xijing Hospital were divided into trial group and control group according to the method of rehabilitation, with 32 cases in each group. Patients in the two groups were given ordinary rehabilitation training for 8 weeks, and the patients in trial group were given a course of group psychological counseling in addition. The Rosenberg's Self-Esteem Scale was used to evaluate the changes in self-confidence levels, and the number of patients with inferiority complex, normal feeling, self-confidence, and over self-confidence were counted before and after treatment. The Abbreviated Burn-Specific Health Scale was used to evaluate physical function, psychological function, social relationship, health condition, and general condition before and after treatment to evaluate the social adaptation of patients. Data were processed with t test, chi-square test, Mann-Whitney U test, and Wilcoxon test., Results: (1) After treatment, the self-confidence levels of patients in trial group were significantly higher than those in control group (Z = -2.573, P < 0.05). Among trial group, the number of patients with inferiority complex was 17 (53.1%) before treatment, which was decreased to 6 (18.8%) after treatment; the number of patients with normal feeling and that of self-confidence were 8 (25.0%) and 4 (12.5%) before treatment, which were respectively increased to 13 (40.6%) and 10 (31.3%) after treatment. The overall difference in trial group was obvious between before and after treatment (Z = -4.123, P < 0.01) . There was no obvious difference in self-confidence level of patients in control group between before and after treatment (Z = -1.000, P > 0.05). (2) After treatment, the scores of psychological function, social relationship, health condition, and general condition were (87 ± 3), (47.8 ± 3.6), (49 ± 3), and (239 ± 10) points in trial group, which were significantly higher than those in control group [(79 ± 4), (38.3 ± 5.6), (46 ± 4), and (231 ± 9) points, with t values respectively -8.635, -8.125, -3.352, -3.609, P values below 0.01]. After treatment, the scores of physical function, psychological function, social relationship, health condition, and general condition in trial group were significantly higher than those before treatment (with t values from -33.282 to -19.515, P values below 0.05). The scores of physical function, psychological function, health condition, and general condition in control group after treatment were significantly higher than those before treatment (with t values from -27.137 to -17.790, P values below 0.05)., Conclusions: Group psychological counseling combined with ordinary rehabilitation training give rise to significant effects on self-confidence level and social adaptation for burn patients.

Objective: To explore the feasibility of IGF2 imprinting system in target gene therapy for tumors., Methods: The mouse H19 enhancer, DMD and promoter H19 were amplified by PCR from mouse genomic DNA and then cloned into the plasmid pDC312. The EGFP and DT-A fragments were amplified by PCR and cloned into the recombinant plasmid, and then the shuttle plasmid were transfected into HEK293 cells together with the adenoviral vector Ad5, namely, Ad-EGFP and Ad-DT-A. Adenovirus hexon gene expression was applied to confirm the presence of adenovirus infections. The effect of the IGF2 imprinting system was tested by fluorescence microscopy. RT-PCR and Western blotting after transfection of the recombinant adenoviral vectors into cancer cells were used to show loss of IGF2 imprinting (LOI) and maintenance of IGF2 imprinting (MOI), respectively. The anti-tumor effect was assessed by MTT and flow cytometry after the HCT-8 (LOI). Human breast cancer cell line MCF-7 (MOI) and human normal gastric epithelial GES-1 (MOI) cell line were transfected with Ad-DT-A in vitro. The anti-tumor effect was detected by injecting the Ad-DT-A in nude mice carrying HCT-8 tumors., Results: The expression of EGFP protein, DT-A mRNA and DT-A protein were seen to be positive only in the HCT-8 tumor cell line. Infection with Ad-DT-A resulted in obviously growth inhibition in HCT-8 cells (75.4 ± 6.4)% compared with that in the control group, and increased the percentage of apoptosis in the HCT-8 cells (20.8 ± 5.9)%. The anti-tumor effect was further confirmed by injecting the recombinant adenoviruses in HCT-8 tumor-bearing nude mice, and the results showed that the Ad-DT-A inhibited the tumor growth, with on inhibition rate of 36.4%., Conclusions: The recombinant adenoviruses carrying IGF2 imprinting system and DT-A gene have been successfully constructed, while Ad-DT-A can effectively kill the tumor cells showing loss of IGF2 imprinting. It might play an important role in future target gene therapy against malignant tumors based on loss of IGF2 imprinting.

CD147, also named extracelluar matrix metalloproteinase inducer (EMMPRIN), is a member of the immunoglobulin family and a glycoprotein enriched on the surface of tumor cells, which promotes invasion, metastasis, growth and survival of malignant cells, and is known to confer resistance to some chemotherapeutic drugs. To determine the possible role of CD147 in the invasive properties of laryngeal carcinoma, we used an RNA interference approach to silence CD147 expression in the Hep2 cell line at high levels of CD147 expression. Our results showed that CD147 expression was significantly impeded at both mRNA and protein levels, which resulted in a decrease of the Hep2 invasion activity in vitro and tumorigenicity in nude mice. The suppression of CD147 expression also sensitized cells to cisplatin. Our current results indicated that CD147 was a laryngeal carcinoma-related gene and CD147 might be a potential target for therapeutic anti-cancer drugs.