Agrin-Lrp4 pathway in hippocampal astrocytes restrains development of temporal lobe epilepsy through adenosine signaling.

BACKGROUND: Human patients often experience an episode of serious seizure activity, such as status epilepticus (SE), prior to the onset of temporal lobe epilepsy (TLE), suggesting that SE can trigger the development of epilepsy. Yet, the underlying mechanisms are not fully understood. The low-density lipoprotein receptor related protein (Lrp4), a receptor for proteoglycan-agrin, has been indicated to modulate seizure susceptibility. However, whether agrin-Lrp4 pathway also plays a role in the development of SE-induced TLE is not clear. METHODS: Lrp4f/f mice were crossed with hGFAP-Cre and Nex-Cre mice to generate brain conditional Lrp4 knockout mice (hGFAP-Lrp4-/-) and pyramidal neuron specific knockout mice (Nex-Lrp4-/-). Lrp4 was specifically knocked down in hippocampal astrocytes by injecting AAV virus carrying hGFAP-Cre into the hippocampus. The effects of agrin-Lrp4 pathway on the development of SE-induced TLE were evaluated on the chronic seizure model generated by injecting kainic acid (KA) into the amygdala. The spontaneous recurrent seizures (SRS) in mice were video monitored. RESULTS: We found that Lrp4 deletion from the brain but not from the pyramidal neurons elevated the seizure threshold and reduced SRS numbers, with no change in the stage or duration of SRS. More importantly, knockdown of Lrp4 in the hippocampal astrocytes after SE induction decreased SRS numbers. In accord, direct injection of agrin into the lateral ventricle of control mice but not mice with Lrp4 deletion in hippocampal astrocytes also increased the SRS numbers. These results indicate a promoting effect of agrin-Lrp4 signaling in hippocampal astrocytes on the development of SE-induced TLE. Last, we observed that knockdown of Lrp4 in hippocampal astrocytes increased the extracellular adenosine levels in the hippocampus 2 weeks after SE induction. Blockade of adenosine A1 receptor in the hippocampus by DPCPX after SE induction diminished the effects of Lrp4 on the development of SE-induced TLE. CONCLUSION: These results demonstrate a promoting role of agrin-Lrp4 signaling in hippocampal astrocytes in the development of SE-induced development of epilepsy through elevating adenosine levels. Targeting agrin-Lrp4 signaling may serve as a potential therapeutic intervention strategy to treat TLE.

PEG-rhG-CSF for prophylaxis of neutropenia after chemotherapy in patients with non-small cell lung cancer: A multicenter, prospective, randomized study.

BACKGROUND: To evaluate the efficacy and safety of pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) in preventing neutropenia during multiple cycles of chemotherapy in patients with non-small cell lung cancer (NSCLC). METHOD: In a multicenter, prospective, randomized trial, patients with NSCLC were randomly assigned in a 2:1 ratio to treatment group (PEG-rhG-CSF as primary prophylactic therapy) or control group. Patients in the control group were administered rhG-CSF when white blood cell count was <2.0 × 109 /L or absolute neutrophil count <1.0 × 109 /L. The primary endpoint was the incidence of grade 3/4 neutropenia. Secondary endpoints included the incidence and duration of grade 3/4 neutropenia in each cycle, the incidence of febrile neutropenia (FN), delay rate of chemotherapy, prolonged time of chemotherapy, and safety. RESULTS: Between January 2019 and July 2021, 130 patients were enrolled (treatment group: n = 87, control group: n = 43). The incidence of grade 3/4 neutropenia in the treatment group was significantly lower than that in the control group (1.15% vs. 11.63%, p < 0.05). The mean duration of grade 3/4 neutropenia for the treatment and control group was 2.00 and 3.75 days, respectively. There were no statistical differences in the incidence of FN, delay rate of chemotherapy, prolonged time of chemotherapy, and antibiotic use between the two groups (all p > 0.05). Adverse events were reported in 47.13% of patients in the treatment group and 48.84% patients in the control group. CONCLUSIONS: Primary prophylactic treatment with PEG-rhG-CSF could reduce the incidence of neutropenia in patients with NSCLC during multiple cycles of chemotherapy, with acceptable safety and tolerability.

Curcumin increases crizotinib sensitivity through the inactivation of autophagy via epigenetic modulation of the miR-142-5p/Ulk1 axis in non-small cell lung cancer.

Drug resistance is a critical factor responsible for the recurrence of non-small cell lung cancer (NSCLC). Previous studies suggest that curcumin acts as a chemosensitizer and radiosensitizer in human malignancies, but the underlying mechanism remains elusive. In the present study, we explored how curcumin regulates the expression of miR-142-5p and sensitizes NSCLC cells to crizotinib. We found that miR-142-5p is significantly downregulated in NSCLC tissue samples and cell lines. Curcumin could increase crizotinib cytotoxicity by epigenetically restoring the expression of miR-142-5p. Furthermore, curcumin treatment suppressed the expression of DNA methylation-related enzymes, including DNMT1, DNMT3A, and DNMT3B, in NSCLC cells. In addition, the upregulation of miR-142-5p expression increased crizotinib cytotoxicity and induced apoptosis in tumor cells in a similar manner to that of curcumin. Strikingly, miR-142-5p overexpression suppressed crizotinib-induced autophagy in A549 and H460 cells. Mechanistically, miR-142-5p inhibited autophagy in lung cancer cells by targeting Ulk1. Overexpression of Ulk1 abrogated the miR-142-5p-induced elevation of crizotinib cytotoxicity in A549 and H460 cells. Collectively, our findings demonstrate that curcumin sensitizes NSCLC cells to crizotinib by inactivating autophagy through the regulation of miR-142-5p and its target Ulk1.

Multilayer network structure enhances the coexistence of competitive species.

The concept of a multiplex network can be used to characterize the dispersal paths and states of different species in a patch habitat system. The multiplex network is one of three types of multilayer networks. In this study, the effect of a multiplex network on the long-term stable coexistence of species is investigated using the concept of metapopulation. Based on the mean field approximation, the stability analysis of a two-species system shows that, compared to the single layer network, the multiplex network is more conducive to the stable coexistence of species when one species has a stronger colonization ability. That is, in such a patch habitat system, if the dispersal paths of the stronger species are different than those of the weaker species, then the larger the heterogeneity of the dispersal network of the stronger species is, the more likely the long-term stable coexistence of species. This result provides a different perspective for understanding the biodiversity in heterogeneous habitats.

Gene methylation as a powerful biomarker for detection and screening of non-small cell lung cancer in blood.

DNA methylation has been reported to become a potential powerful tool for cancer detection and diagnosis. However, the possibilities for the application of blood-based gene methylation as a biomarker for non-small cell lung cancer (NSCLC) detection and screening remain unclear. Hence, we performed this meta-analysis to evaluate the value of gene methylation detected in blood samples as a noninvasive biomarker in NSCLC. A total of 28 genes were analyzed from 37 case-control studies. In the genes with more than three studies, we found that the methylation of P16, RASSF1A, APC, RARß, DAPK, CDH13, and MGMT was significantly associated with risks of NSCLC. The methylation statuses of P16, RASSF1A, APC, RARß, DAPK, CDH13, and MGMT were not linked to age, gender, smoking behavior, and tumor stage and histology in NSCLC. Therefore, the use of the methylation status of P16, RASSF1A, APC, RARß, DAPK, CDH13, and MGMT could become a promising and powerful biomarker for the detection and screening of NSCLC in blood in clinical settings. Further large-scale studies with large sample sizes are necessary to confirm our findings in the future.

Complete Genome Sequence of Vibrio alginolyticus ATCC 17749T.

Vibrio alginolyticus is a Gram-negative halophilic bacterium and has been recognized as an opportunistic pathogen in both humans and marine animals. It is the causative agent of food-borne diseases, such as gastroenteritis, and it invades through wounds in predisposed individuals. In this study, we present the completed genome of V. alginolyticus ATCC 17749(T) through high-throughput sequencing.

Draft Genome Sequence of the Human-Pathogenic Bacterium Vibrio alginolyticus E0666.

Vibrio alginolyticus is a Gram-negative halophilic bacterium with worldwide distribution. In this work, we report the draft genome sequence of a V. alginolyticus strain (E0666) isolated from Epinephelus coioides ascites in the Shantou city of Guangdong Province, China.

Two classifiers based on serum peptide pattern for prediction of HBV-induced liver cirrhosis using MALDI-TOF MS.

Chronic infection with hepatitis B virus (HBV) is associated with the majority of cases of liver cirrhosis (LC) in China. Although liver biopsy is the reference method for evaluation of cirrhosis, it is an invasive procedure with inherent risk. The aim of this study is to discover novel noninvasive specific serum biomarkers for the diagnosis of HBV-induced LC. We performed bead fractionation/MALDI-TOF MS analysis on sera from patients with LC. Thirteen feature peaks which had optimal discriminatory performance were obtained by using support-vector-machine-(SVM-) based strategy. Based on the previous results, five supervised machine learning methods were employed to construct classifiers that discriminated proteomic spectra of patients with HBV-induced LC from those of controls. Here, we describe two novel methods for prediction of HBV-induced LC, termed LC-NB and LC-MLP, respectively. We obtained a sensitivity of 90.9%, a specificity of 94.9%, and overall accuracy of 93.8% on an independent test set. Comparisons with the existing methods showed that LC-NB and LC-MLP held better accuracy. Our study suggests that potential serum biomarkers can be determined for discriminating LC and non-LC cohorts by using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. These two classifiers could be used for clinical practice in HBV-induced LC assessment.

The N-classification for esophageal cancer staging: should it be based on number, distance, or extent of the lymph node metastasis?

BACKGROUND: The recently published AJCC-TNM staging system for esophageal carcinoma made an obvious modification on N-classification based on the number of metastatic regional lymph nodes (LN). However, this classification might ignore the site at which these LNs occur, a factor that might be even more important in reflecting patients' prognosis. METHODS: A retrospective study of 236 patients with carcinoma of thoracic esophagus who underwent esophagectomy between 1984 and 1989 with each at least six LNs removed was conducted, with a 10-year follow-up rate of 92.4%. The proposed scheme for N-classification according to the number (0, 1-2, 3-6, ≥7; N0-3), distance (0, 1, 2, 3 stations; S0-3), or extent (0, 1, and 2 fields; F0-2) of LN involvement was evaluated by univariate and multivariate survival analysis. RESULTS: The LN metastasis was identified in 112 patients, revealing a poorer 5-year survival in this patient group when compared to patients without node involvement. Cox regression analysis revealed that the number and distance of LN metastases and the number of metastasis fields were factors significantly influencing survival. When these factors were further analyzed by univariate log-rank test, no significant difference in survival existed between N2 and N3 patients, or among S1, S2, and S3 patients. When patients were grouped according to the extent of LN metastasis, significant differences in survival were observed overall and between each subgroup. CONCLUSIONS: Refining the current N-classification for esophageal cancer according to the extent of LN metastasis, rather than by number alone, might be a better means of staging that could subgroup patients more effectively and result in different rates of survival.

[Analysis of the survival in patients after surgical resection of thoracic esophageal cancer].

OBJECTIVE: To investigate the prognostic factors and influence of the number of lymph node metastases on survival and UICC-TNM classification in patients with thoracic esophageal cancer after curative resection. METHODS: From 1985 to 1990, 1224 patients were surgically treated for thoracic esophageal cancer. The patients who died within 30 days after operation were not included in this study. Fifteen factors possibly influencing survival of these patients were selected and analyzed. A multivariate analysis of these individual variables was performed by Cox proportional hazard model. According to the number of lymph node metastases (0, 1 and > or = 2), a new modification of the TNM classification was suggested: stage IIa (T2N0M0 and T3N0M0), stage IIb [T1N1M0 and T2N1(1)M0], stage IIIa [T2N1 (2)M0 and T3N1 (1) M0] and stage IIIb [T3N1 (2) M0 and T4N any M0]. RESULTS: According to multivariate analysis, lymph node metastases, depth of invasion, location of tumor, histological classification and length of the tumor were of prognostic significance (P < 0.01). There was obvious correlation between the rate of lymph node metastasis and the depth of invasion, length of tumor and grade of differentiation. The 5-year survival rate of the patients with 0, 1 and > or = 2 positive metastatic lymph nodes was 59.1%, 32.0% and 8. 9%, respectively. The 5-year survival rate of the patients with stage T2N1 M0 and stage T3N1 M0 was significantly higher in those with only one lymph node involved than in those with two or more lymph nodes involved (43.1% vs. 18.0% and 28.0% vs. 9.6%, P < 0.01). The 5-year survival rate of the modified stage IIa, IIb, IIIa and IIIb was 56.5%, 43.9%, 25.6% and 11.1%, respectively, with a statistically significant difference among different stages (P < 0.01). CONCLUSION: The lymph node metastasis is the most important prognostic factor for thoracic esophageal cancer after resection. The major influencing factors of lymph node metastasis are the depth of invasion, length of tumor and grade of differentiation. Therefore, the lymphadenectomy along with esophagectomy and subsequently combined modality therapy against lymph node metastasis is necessary to improve the 5-year survival rate. Our proposed new classification based on number of lymph node metastases (0, 1, > or = 2 positive nodes) is more applicable because it can well reflect the correlation between lymph node metastasis and the survival, and provides evidence for the modification of the currently used UICC TNM staging system for surgically treated thoracic esophageal cancer.

[Influence of number of removed lymph nodes on the TNM staging and survival in advanced esophageal carcinoma].

OBJECTIVE: To evaluate the impact of total number of removed regional lymph nodes (LN) during esophagectomy on TNM staging and long-term survival. METHODS: Between 1984 and 1989, a total of 1098 patients with advanced squamous cell carcinoma of the thoracic esophagus were surgically treated, and the data were retrospectively analyzed. The survival was analyzed by Kaplan-Meier method. Multivariate and univariate analysis were performed using Cox proportional hazard model and Log-rank test respectively in order to compare the survival difference and the impact of <6 or > or =6 removed LN on TNM staging and survival. RESULTS: The shortest follow-up time was 10 years with a follow-up rate of 91.4%. Of these 1098 patients, 378 patients were found to have regional LN metastases with a LN metastasis rate of 34.4%. Totally, 4157 LN were removed and tumor spreading into the lymph node was documented in 800 with a lymph node metastasis degree of 19.2%. The mean number of removed LN in each esophgectomy was 3.8. According to the number of total removed LN, patients were divided into group A (removed LN <6, N=825) and group B (removed LN > or =6, N=273). The survival of group A was worse than that of group B. The depth of tumor invasion, degree of lymph node metastasis, tumor location, and tumor residual status were the most important prognostic factors. Even though the lymphatic metastasis rate in group A was significantly lower than that in group B (30.3% vs. 46.9% P < 0.001), the LN metastasis degree was higher in group A than that in group B (21.2% vs. 17.5% P = 0.002) due to less number of removed LN in group A than in group B. With the stratification analysis according to the number of involved lymph nodes, for patients without LN spreading, the survival in group A was significantly worse than that in group B (P = 0.003), but in the patients with only one or > or =2 positive nodes, the survival was similar (P = 0.919 and 0.182, respectively). When stratified patients according to TNM stage, for stage IIa patients (T2N0M0, T3N0M0), the survival in group A was significantly worse than that in group B (P = 0.006), while such difference did not exist in patients with stage IIb or stage lIII(P = 0.302 and 0.108, respectively). CONCLUSION: A large series of retrospective study on advanced squamous cell carcinoma of the thoracic esophagus demonstrates that the number of metastatic LN is an important prognostic factor, therefore, it should be considered when refining UICC-TNM classification for esophageal cancer. If the total number of LN removed during each esophagectomy is less than 6, the occult positive regional LN might be missed, resulting in an inaccurate N classification and incorrect staging. Removal of > or = LN for each esophageal cancer patient during esophagectomy recommended by UICC is rational and should be complied with.