Xanthomonas oryzae pv. oryzicola effector Tal10a directly activates rice OsHXK5 expression to facilitate pathogenesis.

Bacterial leaf streak (BLS), caused by Xanthomonas oryzae pv. oryzicola (Xoc), is a major bacterial disease in rice. Transcription activator-like effectors (TALEs) from Xanthomonas can induce host susceptibility (S) genes and facilitate infection. However, knowledge of the function of Xoc TALEs in promoting bacterial virulence is limited. In this study, we demonstrated the importance of Tal10a for the full virulence of Xoc. Through computational prediction and gene expression analysis, we identified the hexokinase gene OsHXK5 as a host target of Tal10a. Tal10a directly binds to the gene promoter region and activates the expression of OsHXK5. CRISPR/Cas9-mediated gene editing in the effector binding element (EBE) of OsHXK5 significantly increases rice resistance to Xoc, while OsHXK5 overexpression enhances the susceptibility of rice plants and impairs rice defense responses. Moreover, simultaneous editing of the promoters of OsSULTR3;6 and OsHXK5 confers robust resistance to Xoc in rice. Taken together, our findings highlight the role of Tal10a in targeting OsHXK5 to promote infection and suggest that OsHXK5 represents a potential target for engineering rice resistance to Xoc.

High-Frequency and High-Current Transmission Techniques for Multiple Earth Electrical Characteristic Measurement Systems Based on Adaptive Impedance Matching through Phase Comparison.

With the increase in groundwater exploration, underground mineral resource exploration, and non-destructive investigation of cultural relics, high-resolution earth electrical characteristic measurement has emerged as a mainstream technique owing to its advantageous non-destructive detection capability. To enhance the transmission power of the high-frequency transmitter in high-resolution multiple earth electrical characteristic measurement systems (MECS), this study proposes a high-frequency, high-current transmission technique based on adaptive impedance matching and implemented through the integration of resonant capacitors, a controllable reactor, high-frequency transformers, and corresponding control circuits. A high-current precisely controllable reactor with a 94% inductance variation range was designed and combined with resonant capacitors to reduce circuit impedance. Additionally, high-frequency transformers were employed to further increase the transmission voltage. A prototype was developed and tested, demonstrating an increase in transmission current at frequencies between 10 and 120 kHz with a peak active power of 200 W. Under the same transmission voltage, compared to the transmission circuit without impedance matching, the transmission current increased to a maximum of 16.7 times (average of 10.8 times), whereas compared to the transmission circuit using only traditional impedance matching, the transmission current increased by a maximum of 10.0 times (average of 4.2 times), effectively improving the exploration resolution.

A Piezoelectric-Piezoresistive Coupling Electric Field Sensor for Large Dynamic Range AC Electric Field Measurements.

We propose a piezoelectric-piezoresistive coupling electric field sensor capable of performing large dynamic range AC electric field measurements. The electric field sensor utilizes direct coupling between piezoelectric (PE) materials and piezoresistive (PR) strain gauges, in conjunction with an external signal conditioning circuit, to measure AC electric fields effectively. We verified the feasibility of the scheme using a finite element simulation, fabricated a prototype of the electric field sensor, and characterized the properties of the prototype. The testing results indicate that the sensor exhibits an ac resolution of 50 V/m and a linear measurable electric field range of 0 to over 200 kV/m, which keeps the linearity at less than 0.94% from 1 Hz to over 5 kHz. Furthermore, the sensor also has advantages, such as a small size and low power consumption. The sensor can enhance the comprehensive observability and measurability of digital power grids.

VmsR, a LuxR-Type Regulator, Contributes to Virulence, Cell Motility, Extracellular Polysaccharide Production and Biofilm Formation in Xanthomonas oryzae pv. oryzicola.

LuxR-type regulators play pivotal roles in regulating numerous bacterial processes, including bacterial motility and virulence, thereby exerting a significant influence on bacterial behavior and pathogenicity. Xanthomonas oryzae pv. oryzicola, a rice pathogen, causes bacterial leaf streak. Our research has identified VmsR, which is a response regulator of the two-component system (TCS) that belongs to the LuxR family. These findings of the experiment reveal that VmsR plays a crucial role in regulating pathogenicity, motility, biofilm formation, and the production of extracellular polysaccharides (EPSs) in Xoc GX01. Notably, our study shows that the vmsR mutant exhibits a reduced swimming motility but an enhanced swarming motility. Furthermore, this mutant displays decreased virulence while significantly increasing EPS production and biofilm formation. We have uncovered that VmsR directly interacts with the promoter regions of fliC and fliS, promoting their expression. In contrast, VmsR specifically binds to the promoter of gumB, resulting in its downregulation. These findings indicate that the knockout of vmsR has profound effects on virulence, motility, biofilm formation, and EPS production in Xoc GX01, providing insights into the intricate regulatory network of Xoc.

Therapeutic effects of Buyang Huanwu Tang combined with RT-PA intravenous thrombolysis on stroke of Qi deficiency and blood stasis type and its impact on Keap1-Nrf2/ARE pathway antioxidant stress.

This study investigated the impact of combining traditional Chinese medicine, Buyang Huanwu Tang, with intravenous thrombolysis using alteplase (rt-PA) in treating ischemic stroke patients with qi deficiency and blood stasis. A single-center clinical randomized trial involved 117 ischemic stroke patients treated with rt-PA in the neurology department from January 2019 to December 2021. Patients were randomly divided into two groups: the control group (58 patients) received rt-PA alone, while the combined group (59 patients) received rt-PA along with Buyang Huanwu Tang. Neurological deficit scores (NIHSS) were assessed before and after treatment, along with hemorheological indicators, vascular endothelial growth factor (VEGF), matrix metalloproteinase-9 (MMP-9), and Keap1-Nrf2/ARE pathway oxidative stress indicators (Keap1, Nrf2, ARE, and NQO1 proteins). Before treatment, there were no significant differences between the groups. After treatment, the combination group exhibited lower NIHSS scores at 4, 8, and 12 weeks, indicating significant improvement compared to the control group. Additionally, the combination group demonstrated reduced plasma viscosity, low and high shear viscosity, and improved red blood cell aggregation compared to the control group after 8 weeks of treatment. Furthermore, the combination group showed elevated MMP-9 levels and reduced VEGF levels, suggesting favorable outcomes. Regarding the Keap1-Nrf2/ARE pathway, Nrf2 and NQO1 protein expression levels were higher in the combination group after 8 weeks of treatment. Clinical efficacy assessment revealed that the combined treatment group had a significantly better overall treatment response. In conclusion, combining Buyang Huanwu Tang with rt-PA intravenous thrombolysis effectively mitigated oxidative stress damage in the Keap1-Nrf2/ARE pathway among ischemic stroke patients with qi deficiency and blood stasis. This approach promoted neurological function recovery and improved overall treatment outcomes.

Comparative analysis of the clinical and epidemiological characteristics of human influenza virus versus human respiratory syncytial virus versus human metapneumovirus infection in nine provinces of China during 2009-2021.

A comparative analysis of confirmed cases of human influenza virus (HIFV), human respiratory syncytial virus (HRSV), and human metapneumovirus (HMPV) was conducted to describe their clinical and epidemiological characteristics. During 2009-2021, active surveillance of acute respiratory infections (ARIs) was performed in nine provinces of China. Clinical and epidemiological information and laboratory testing results of HIFV, HRSV, and HMPV were analyzed. Among 11591 ARI patients, the single-infection rates of HIFV, HRSV, and HMPV were 15.00%, 9.59%, and 2.24%, respectively; the coinfection rate of these three viruses was 0.64%. HIFV infection was mainly in adults aged 15-59 years, accounting for 39.10%. HRSV and HMPV infections were mainly in children under 5 years old, accounting for 87.13% and 83.46%, respectively. Patients with HRSV infection were younger than HMPV. HRSV and HMPV had high similarities in clinical manifestations, presenting with lower respiratory symptoms. HIFV mainly presented with an upper respiratory infection. The epidemic peak of HRSV was earlier than that of HIFV, and that of HMPV was later than those of HRSV and HFIV. A total of 85.14% of coinfection cases were children under 5 years old. Coinfection might increase the risk of pneumonia in HIFV cases. During 2020-2021, the positive rates and seasonal patterns of these three viruses changed due to the impact of the COVID-19 pandemic. Certain clinical and epidemiological features were observed in HIFV, HRSV, and HMPV infections, which could be beneficial for guiding clinical diagnosis, treatment, and prevention of these three viruses in China.

[Protective effect of curcumin on dopamine neurons in Parkinson's disease and its mechanism].

OBJECTIVE: To investigate the effect of curcumin on dopamine neurons in Parkinson's disease (PD) and its mechanism. METHODS: SH-SY5Y human neuroblastoma cells were treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to establish the PD cell model. The model cells were treated with curcumin and/or autophagy inhibitor 3-MA. After 48 h of drug treatment, the number of surviving dopamine neurons was detected by tyrosine hydroxylase immunofluorescence method. Western blotting was used to detect protein expression of α-Synuclein (α-Syn), transcription factor EB (TFEB) and autophagy-related proteins lysosome-associated membrane protein 2A (LAMP2A) and microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ); RT-PCR was used to detect mRNA expression of α-Syn. RESULTS: Compared with MPTP model group, curcumin increased the number of surviving dopamine neurons(P<0.01), decreased both protein expression and mRNA expression of α-Syn (all P<0.01), and increased protein expression of TFEB, LAMP2A and LC3-Ⅱ (all P<0.01). When curcumin and 3-MA were given concurrently, the number of surviving dopamine neurons, protein expression of TFEB, LAMP2A and LC3-Ⅱ increased (P<0.05 or P<0.01), and both protein expression and mRNA expression of α-Syn decreased (P<0.05 or P<0.01) compared with MPTP model group; but the number of surviving dopamine neurons and protein expression of LAMP2A and LC3-Ⅱ decreased compared with curcumin group (all P<0.05). CONCLUSIONS: Curcumin exerts protective effect on dopamine neurons in PD, which may be associated with enhancing autophagy and promoting the clearance of α-Syn.

Highly sensitive detection of cancer-related genes based on complete fluorescence restoration of a molecular beacon with a functional overhang.

The accurate detection of cancer-related genes is of great significance for early diagnosis and targeted therapy of cancer. In this contribution, an automatically cycling operation of a functional overhang-containing molecular beacon (OMB)-based sensing system was proposed to perform amplification detection of the p53 gene. Contrary to the common molecular beacon (MB), a target DNA is designated to hybridize with a label-free recognition probe (RP) with a hairpin structure rather than OMB. In the presence of a target DNA of interest, the locked primer in RP opens and triggers the subsequent amplification procedures. The newly-developed OMB is not only capable of accomplishing cyclical nucleic acid strand-displacement polymerization (CNDP) with the help of polymerase and nicking endonuclease, but is also cleaved by restriction endonucleases, removing the quencher away from the fluorophore. Thus, the target DNA at an extremely low concentration is expected to generate a considerable amount of double-stranded and cleaved OMBs, and the quenched fluorescence is completely restored, leading to a dramatic increase in fluorescence intensity. Utilizing this sensing platform, the target gene can be detected down to 8.2 pM in a homogeneous way, and a linear response range of 0.01 to 150 nM could be obtained. More strikingly, the mutant genes can be easily distinguished from the wild-type ones. The proof-of-concept demonstrations reported herein are expected to promote the development of DNA biosensing systems, showing great potential in basic research and clinical diagnosis.

Double-hairpin molecular-beacon-based amplification detection for gene diagnosis linked to cancer.

A powerful double-hairpin molecular beacon (DHMB) was developed for cancer-related KRAS gene detection based on the one-to-two stoichiometry. During target DNA detection, DHMB can execute signal transduction even if no any exogenous element is involved. Unlike the conventional molecular beacon based on the one-to-one interaction, one target DNA not only hybridizes with one DHMB and opens its hairpin but also promotes the interaction between two DHMBs, causing the separation of two fluorophores from quenchers. This leads to an enhanced fluorescence signal. As a result, the target KRAS gene is able to be detected within a wide dynamic range from 0.05 to 200 nM with the detection limit of 50 pM, indicating a dramatic improvement compared with traditional molecular beacons. Moreover, the point mutations existing in target DNAs can be easily screened. The potential application for target species in real samples was indicated by the analysis of PCR amplicons of DNAs from the DNA extracted from SW620 cell. Besides becoming a promising candidate probe for molecular biology research and clinical diagnosis of genetic diseases, the DHMB is expected to provide a significant insight into the design of DNA probe-based homogenous sensing systems. Graphical Abstract A powerful double-hairpin molecular beacon (DHMB) was developed for cancer-related gene KRAS detection based on the one-to-two stoichiometry. Without the help of any exogenous probe, the point mutation is easily screened, and the target DNA can be quantified down to 50 pM, indicating a dramatic improvement compared with traditional molecular beacons.

Autophagy regulation revealed by SapM-induced block of autophagosome-lysosome fusion via binding RAB7.

The mechanism underlying autophagy alteration by mycobacterium tuberculosis remains unclear. Our previous study shows LpqH, a lipoprotein of mycobacterium tuberculosis, can cause autophagosomes accumulation in murine macrophages. It is well known that SapM, another virulence factor, plays an important role in blocking phagosome-endosome fusion. However, the mechanism that SapM interferes with autophagy remains poorly defined. In this study, we report that SapM suppresses the autophagy flux by blocking autophagosome fusion with lysosome. Exposure to SapM results in accumulations of autophagosomes and decreased co-localization of autophagosome with lysosome. Molecularly, Rab7, a small GTPase, is blocked by SapM through its CT domain and is prevented from involvement of autophagosome-lysosome fusion. In conclusion, our study reveals that SapM takes Rab7 as a previously unknown target to govern a distinct molecular mechanism underlying autophagosome-lysosome fusion, which may bring light to a new thought about developing potential drugs or vaccines against tuberculosis.

Effect of site-specific PEGylation on the fibrinolytic activity, immunogenicity, and pharmacokinetics of staphylokinase.

The bacterial plasminogen-activator staphylokinase (Sak) is a promising thrombolytic agent for treating acute myocardial infarction. To effectively reduce the immunogenicity of Sak while maintaining its fibrinolytic activity, site-specific PEGylation was performed in the present study. The chemoselective cysteine PEGylation site was selected within an immunodominant region (amino acid residues 71-87) using an in silico approach. The PEGylated Sak variants prepared in this study showed a purity of >97.0%. PEGylation at Position 80 resulted in a Sak variant Sak(E80C-PEG) which has similar fibrinolytic activity and thermostability compared with the native recombinant staphylokinase (r-Sak). The immunogenicity of Sak(E80C-PEG) in guinea pigs was greatly reduced compared with the native r-Sak. Furthermore, preliminary pharmacokinetic results suggested that the plasma clearance of Sak(E80C-PEG) from the blood stream of rabbit was significantly decreased compared with that of r-Sak, resulting in a 2.8-fold increase of initial half-life and a 3.8-fold increase of systemic availability. In summary, these results demonstrated that site-specific PEGylation yielded a novel Sak variant Sak(E80C-PEG) with remarkable advantages over the unmodified Sak.

Innovation driver and county air pollution: cost-benefit analysis perspective.

Innovation is the first power to drive the county's green and low-carbon. It is crucial to explore the impact of innovation on air pollution from the perspective of counties at the bottom of the administrative division hierarchy. The article is aimed at exploring the direct impact effects, spatial spillover effects, impact mechanism pathways, non-linear relationships, and cost-benefits of innovation drive on air pollution in counties. To this end, based on the collection of county-level data from 2007 to 2020 in mainland China, the article constructs a fixed-effects model, a dynamic panel model, and a spatial Durbin model for analysis. For every 1% increase in the quantity of innovation, the county SO2 emission concentration decreases by 0.2% on average; for every 1% increase in the quality of innovation, the county SO2 emission concentration decreases by 0.3% on average. When the county innovation quantity driver increases by one standard deviation, the county SO2 concentration decreases by an average of 0.29%; when the county innovation quality driver each standard deviation increases, the county SO2 concentration is reduced by 0.33% on average. The significant entry of high-end factors, the increased frequency of regulation by the environmental protection department, and the increasing efficiency of energy use are the important mechanism pathways for innovation-driven reduction of air pollution in counties. There is no significant "(inverted) U-shaped" relationship between innovation-driven air pollution in the county samples. There is a negative spatial spillover effect of the innovation quality drive on air pollution control in all Chinese county samples. Innovation to drive the declining size of the county's sulfur dioxide can bring about one billion yuan (about 139.81 million U.S. dollars) in comprehensive economic benefits. In the coming period, county governments should build a new pattern of "blue sky and white clouds" with neighboring regions in terms of spatial agglomeration of high-end elements, green transformation and utilization of energy, and intelligent monitoring and supervision of pollution.

Study on pyroptosis-related genes Casp8, Gsdmd and Trem2 in mice with cerebral infarction.

Objective: Cerebral infarction is the main cause of death in patients with cerebrovascular diseases. Our research aimed to screen and validate pyroptosis-related genes in cerebral infarction for the targeted therapy of cerebral infarction. Methods and results: A total of 1,517 differentially expressed genes (DEGs) were obtained by DESeq2 software analysis. Gene set enrichment analysis results indicated that genes of middle cerebral artery occlusion (MCAO) mice aged 3 months and 18 months were enriched in pyroptosis, respectively. Differentially expressed pyroptosis-related genes (including Aim2, Casp8, Gsdmd, Naip2, Naip5, Naip6 and Trem2) were obtained through intersection of DEGs and genes from pyroptosis Gene Ontology Term (GO:0070269), and they were up-regulated in the brain tissues of MCAO mice in GSE137482. In addition, Casp8, Gsdmd, and Trem2 were verified to be significantly up-regulated in MCAO mice in GSE93376. The evaluation of neurologic function and triphenyltetrazolium chloride staining showed that the MCAO mouse models were successfully constructed. Meanwhile, the expressions of TNF-α, pyroptosis-related proteins, Casp8, Gsdmd and Trem2 in MCAO mice were significantly up-regulated. We selected Trem2 for subsequent functional analysis. OGD treatment of BV2 cell in vitro significantly upregulated the expressions of Trem2. Subsequent downregulation of Trem2 expression in OGD-BV2 cells further increased the level of pyroptosis. Therefore, Trem2 is a protective factor regulating pyroptosis, thus influencing the progression of cerebral infarction. Conclusions: Casp8, Gsdmd and Trem2 can regulate pyroptosis, thus affecting cerebral infarction.

Dimeric Pillar[5]arene as a Novel Fluorescent Host for Controllable Fabrication of Supramolecular Assemblies and Their Photocatalytic Applications.

A dimeric fluorescent macrocycle m-TPE Di-EtP5 (meso-tetraphenylethylene dimeric ethoxypillar[5]arene) is synthesized based on the meso-functionalized ethoxy pillar[5]arene. Through the connectivity of two pillar[5]arenes by CC double bond, the central tetraphenylethylene (TPE) moiety is simultaneously formed. The resultant bicyclic molecule not only retains the host-guest properties of pillararenes but also introduces the interesting aggregation-induced emission properties inherent in the embedded TPE structure. Three dinitrile derivatives with various linkers are designed as guests (G1, G2, and G3) to form host-guest assemblies with m-TPE Di-EtP5. The morphological control and fluorescence properties of the assemblies are successfully realized. G1 with a shorter alkyl chain as the linker completely threads into the cavities of the host. G2, due to its longer chain length, forms a linear supramolecular polymer upon binding to m-TPE Di-EtP5. G3 differs from G2 by possessing a bulky phenyl group in the middle of the chain, which can be further assembled with m-TPE Di-EtP5 to form supramolecular layered polymer and precipitated out in solution, and can be efficiently applied to photocatalytic reactions.

p53 and p73 expression in esophageal carcinoma correlate with clinicopathology of tumors.

BACKGROUND/AIMS: Inactivation of tumor suppressors like p53 and p73 plays an important role in cancer initiation. We explored the clinical significance of p53 and p73 expression in esophageal carcinoma, along with their correlation with clinicopathological parameters of tumors for potential use in clinical evaluation of this disease. METHODOLOGY: In the present study, tumor samples and adjacent normal tissue samples were collected from 37 patients with esophageal cancer and the expression of p53 and p73 was detected by immunohistochemistry and correlated with clinicopathological parameters of tumors. RESULTS: p53 and p73 were more frequently expressed in esophageal cancer than in adjacent normal tissue. Indeed, there was a positive correlation between p53 and p73 expression and esophageal cancer (p<0.05). Analysis of clinicopathological features revealed that p53 expression was correlated with differentiation, distant metastases, and TNM stage (p<0.05) of tumors, but not with gender; age, infiltration depth, tumor size, or lymph node metastasis. In contrast, p73 expression was correlated only with distant metastases (p<0.05). CONCLUSIONS: Increased expression of p53 and p73 in esophageal carcinoma may serve as an indicator of tumor severity. Detection of these proteins in future studies may help understand the mechanisms of development, invasion and metastasis in esophageal cancer.

Can the Adjustment and Renovation Policies of Old Industrial Cities Reduce Urban Carbon Emissions?-Empirical Analysis Based on Quasi-Natural Experiments.

Based on a literature review and theoretical mechanism, this paper takes the implementation point of the adjustment and transformation policy for old industrial cities as the breakthrough point, and uses a regression model to explore the impact of the adjustment and transformation policy of these old industrial cities on urban carbon emissions. This paper also robustly tests the effective mechanisms and environmental hypotheses. Overall, the implementation of the adjustment and renovation policy has significantly reduced the carbon emissions of old industrial cities by about 0.068 units. Compared with the control group cities, the pilot cities reduced carbon emissions by an average of about 310,000 tons after the implementation of the policy. Based on a summary of the excellent Chinese case experience and an empirical analysis, it can be concluded that improvements in the green innovation capacity of old industrial cities, the agglomeration of high-end service industries, and the strengthening of ecological restoration are important mechanisms that lead to reduced carbon emissions. There is no subsequent exacerbation of the carbon intensity of neighboring cities, and there is insufficient evidence to prove pollution via neighboring transfers and use of the beggar-thy-neighbor policy. The extended analysis shows that the "inverted U-shaped" CO2 Kuznets environmental curve hypothesis is significantly present in the sample of old industrial cities, but most cities do not cross the threshold. In 2013, about 60% of the urban sample economic growth and carbon emissions showed signs of tapping into potentials and increasing efficiency (absolute decoupling) and intensive expansion (relative decoupling). In old industrial cities, the proportion of relative decoupling shows a fluctuating upward trend. In the future, the government should accurately select its own development orientation and actively seek the "best balance" between economic growth and a green and low-carbon path.

[Effect of epitope-based peptide-DNA dual vaccines against Schistosoma japonicum in mice].

A C-T-B PDDV mixture of the three constructed epitope-based peptide-DNA dual vaccines (PDDV) containing the CTL (C), Th (T) and B-cell (B) epitopes from Sj22.6 tegument (C-PDDV, T-PDDV and B-PDDV) with a 1:1:1 ratio was prepared. Thirty-six mice were randomly divided into six groups averagely named as 18K group, PBS group, C-PDDV group, T-PDDV group, B-PDDV group, and C-T-B PDDV group. All the mice received three immunizations at 2-week intervals with the same dose of antigen (10 microg DNA+28 microg peptide). One week after the last immunization, the mice were sacrificed, the spleens were removed and splenocytes were collected. Splenocyte proliferation was assayed by[3H] TdR incorporation after stimulation with soluble worm antigen (SWA). Levels of IFN-gamma and IL-4 in the splenocyte culture supernatants were determined by ELISA. The results showed that IFN-gamma content in T-PDDV group [(76.0 +/- 11.2) pg/ml] was higher than that of PBS [(13.0 +/- 2.1) pg/ml] and 18K control groups [(14.0 +/- 3.2) pg/ml] (P<0.01). IL-4 level in T-PDDV [(152.0 +/- 21.1) pg/ml] and C-T-B mixture groups [(86.0 +/- 12.2) pg/ml] was higher than others (P<0.01 and P<0.05). The splenocytes from T-PDDV group showed a significant increase in proliferation compared with PBS and 18K control groups after stimulation by SWA (P<0.01). However, there was no significant difference in splenocyte proliferation among C-T-B, PBS and 18K control groups (P>0.05). These findings indicate that T-PDDV and C-T-B PDDV mixture induces stronger immune response than that of C-PDDV or B-PDDV.

Association of Multiple Dopamine D3 Receptor Gene 3'UTR Polymorphisms with Susceptibility to Parkinson's Disease and Clinical Efficacy of Piribedil Therapy.

Objective: To investigate the correlation between the Dopamine D3 receptor (DRD3) 3'untranslated region (3'UTR) gene polymorphism and susceptibility to Parkinson's disease (PD) and the clinical effect of the DRD2 and DRD3 agonist piribedil treatment. Methods: Sanger sequencing was used to analyze the single nucleotide polymorphisms (SNPs) within the 3'UTR rs76126170, rs9868039, rs9817063, and rs3732790 loci of the DRD3 gene in 284 PD patients and 284 controls. PD patients were treated with piribedil sustained-release tablets (50 mg) combined with levodopa and benserazide hydrochloride tablets, three times daily (patients with first-diagnosed PD were only administrated with piribedil sustained-release tablets) for 3 months. The Unified Parkinson's Disease Rating Scale (UPDRS) and the Hoehn and Yahr disease stage were evaluated at baseline and after 3 months of treatment. Results: The T allele carriers of the DRD3 gene rs76126170 locus were more susceptible to PD than the C allele carriers (odds ratio [OR] = 3.44, 95% confidence interval [CI]: 2.46-4.80, p < 0.01). Carriers of the rs9868039 A allele had a decreased risk of PD compared to those with G allele (OR = 0.67, 95% CI: 0.53-0.86, p < 0.01). C allele carriers at rs9817063 were less likely to develop PD than those with T allele (OR = 0.74, 95% CI: 0.58-0.94, p = 0.02). No significant correlation was observed between the alleles or genotypes of the rs3732790 locus and PD susceptibility (p > 0.05). The various genotypes of the DRD3 gene loci rs76126170, rs9868039, and rs9817063 in PD patients were associated with significant differences with regard to reduction of UPDRS scores and Hoehn and Yahr stage after 3 months of treatment (p < 0.05). Conclusion: The alleles and genotypes of the DRD3 gene 3' UTR SNP loci rs76126170, rs9868039, and rs9817063 are associated with PD susceptibility and the clinical efficacy of piribedil treatment.

Cornuside alleviates experimental autoimmune encephalomyelitis by inhibiting Th17 cell infiltration into the central nervous system.

The present study was conducted to clarify the therapeutic effect of cornuside on experimental autoimmune encephalomyelitis (EAE) and its influence on T helper 17 (Th17) cell and regulatory T (Treg) cell infiltration into the central nervous system. Rats were randomly placed into four treatment groups: control, EAE, EAE+cornuside, and EAE+prednisolone. The neurological function scores of rats were assessed daily. On the second day after EAE rats began to show neurological deficit symptoms, the four groups were treated with normal saline, normal saline, cornuside (150 mg/kg), and prednisolone (5 mg/kg), respectively. The treatment was discontinued after two weeks, and the spinal cord was obtained for hematoxylin and eosin (H&E) and luxol fast blue staining, as well as retinoic acid receptor-related orphan receptor γ (RORγ) and forkhead box protein P3 (Foxp3) immunohistochemical staining. Blood was collected for Th17 and Treg cell flow cytometry testing, and the serum levels of interleukin (IL)-17A, IL-10, transforming growth factor-ß (TGF-ß), IL-6, IL-23, and IL-2 were measured via enzyme-linked immunosorbent assay (ELISA). Compared with rats in the EAE group, rats in the EAE+cornuside and EAE+prednisolone groups began to recover from neurological deficits earlier, and had a greater degree of improvement of symptoms. Focal inflammation, demyelination, and RORγ-positive cell infiltration were reduced by cornuside or prednisolone treatment, whereas the Foxp3-positive cell numbers were not significantly different. Meanwhile, the number of Th17 cells and the IL-17A, IL-6, and IL-23 levels were lower in the blood after cornuside or prednisolone treatment, whereas the number of Treg cells or the levels of IL-10, TGF-ß, and IL-2 were not markedly different. Cornuside can alleviate symptoms of EAE neurological deficits through its anti-inflammatory and immunosuppressive effects, and Th17 cells may be one of its therapeutic targets.

Gefitinib sensitization of cisplatin-resistant wild-type EGFR non-small cell lung cancer cells.

PURPOSE: The usual first-line strategy of wild-type EGFR (wtEGFR) non-small cell lung cancer (NSCLC) remains cisplatin-based chemotherapy. However, cisplatin often loses effectiveness because most tumors acquire drug resistance over time. As EGFR is the most important pro-survival/proliferation signal receptor in NSCLC cells, we aimed at investigating whether cisplatin resistance is related to EGFR activation and further evaluating the combined effects of cisplatin/gefitinib (EGFR-tyrosine kinase inhibitor, EGFR-TKI) on cisplatin-resistant wtEGFR NSCLC cells. MATERIALS AND METHODS: EGFR activation was analysed in parental and cisplatin-resistant wtEGFR NSCLC cell lines (H358 and H358R, A549 and A549R). Cellular proliferation and apoptosis of H358R/A549R cells treated with cisplatin or gefitinib, alone or in combination were investigated, and the related effector protein was detected by western blot analysis. Anti-tumor effect of two drugs combined was evaluated in animal models of H358R xenografts in vivo. RESULTS: EGFR was significantly phosphorylated in cisplatin-resistant wtEGFR NSCLC cells H358R and A549R than their parental cells. In H358R and A549R cells, anti-proliferative ability of gefitinib was further improved, and gefitinib combined with cisplatin enhanced inhibition of cellular survive/proliferation, and promotion of apoptosis in vitro. The combined effects were also associated with the inhibition of EGFR downstream effector proteins. Similarly, in vivo, gefitinib and cisplatin in combination significantly inhibited tumor growth of H358R xenografts. CONCLUSION: Abnormal activation of EGFR may induce wtEGFR NSCLC cell resistance to cisplatin. The combined effects of cisplatin/gefitinib suggest that gefitinib, as a combination therapy for patients with cisplatin-resistant wtEGFR NSCLC should be considered.