Distinct roles of the Hox genes Ultrabithorax and abdominal-A in scorpionfly embryonic proleg development.

The abdominal appendages of larval insects have a complex evolutionary history of gain and loss, but the regulatory mechanisms underlying the abdominal appendage development remain largely unclear. Here, we investigated the embryogenesis of abdominal prolegs in the scorpionfly Panorpa liui Hua (Mecoptera: Panorpidae) using in situ hybridization and parental RNA interference. The results show that RNAi-mediated knockdown of Ultrabithorax (Ubx) led to a homeotic transformation of the first abdominal segment (A1) into the third thoracic segment (T3) and changed the distributions of the downstream target Distal-less (Dll) expression but did not affect the expression levels of Dll. Knockdown of abdominal-A (abd-A) resulted in malformed segments, abnormal prolegs and disrupted Dll expression. The results demonstrate that the gene Ubx maintains an ancestral role of modulating A1 appendage fate without preventing Dll initiation, and a secondary adaptation of abd-A evolves the ability to specify abdominal segments and proleg identity. We conclude that changes in abdominal Hox gene expression and their target genes regulate abdominal appendage morphology during the evolutionary course of holometabolous larvae.

Copepods as Indicators of Different Water Masses during the Northeast Monsoon Prevailing Period in the Northeast Taiwan.

During this research, the average surface temperature, salinity, dissolved oxygen, and pH were 24.65 ± 1.53 (°C), 34.21 ± 0.07 (PSU), 6.85 ± 0.18 (mg/L), and 8.36 ± 0.03, respectively. Based on these environmental parameters, stations were arranged into three groups. Group A represents stations located around Keelung Island with the relative highest average dissolved oxygen, lowest average temperature, and pH values. Instead, the lowest average dissolved oxygen and highest average temperature, salinity, and pH values were recorded at the offshore stations. Keelung Island area was charged by cold water masses, which were driven by the Northeast monsoon, and stations in group C were affected by the Kuroshio Current. Kueishan Island area was mainly affected by mixed water masses resulting from the Kuroshio intrusion and monsoon-derived cold water. In this study, a total of 108 copepod species were identified, with an average abundance of 774.24 ± 289.42 (inds. m-3). Most species belong to the orders Calanoida and Poecilostomatoida, with an average relative abundance (RA) of 62.96% and 30.56%, respectively. Calanoid copepodites were the most dominant group, with a RA of 28.06%. This was followed by Paracalanus aculeatus, with a RA of 18.44%. The RA of Clausocalanus furcatus and Canthocalanus pauper was 4.80% and 3.59%, respectively. The dominant species P. aculeatus, C. pauper, Paracalanus parvus, and Temora turbinata were positively correlated with dissolved oxygen and negatively correlated with temperature in the surface waters. pH showed a negative correlation with P. parvus and T. turbinata, while the temperature was negatively correlated with these two dominant species. Indicator species were selected by an indicator value higher than 50%. Temora turbinata, Calanopia elliptica, C. pauper, Euchaeta concinna, Temora discaudata, Acartia pacifica, Macrosetella gracilis, Corycaeus speciosus, and P. parvus were considered as monsoonal cold water indicator species in Group A. Indicator copepod species for the Kuroshio Current were Farranula concinna, Copilia mirabilis, Candacia aethiopica, Corycaeus agilis, Farranula gibbula and Acrocalanus monachus in the study area. Paracandacia truncata, Oncaea clevei, P. aculeatus, and Centropages furcatus were considered suitable indicators for mixed water masses.

The Hox gene Abdominal-B regulates the appendage development during the embryogenesis of scorpionflies.

The Homeotic Complex (Hox) genes encode conserved homeodomain transcription factors that specify segment identity and appendage morphology along the antero-posterior axis in bilaterian animals. The Hox gene Abdominal-B (Abd-B) is mainly expressed in the posterior segments of the abdomen and plays an important role in insect organogenesis. In Mecoptera, the potential function of this gene remains unclear yet. Here, we performed a de novo transcriptome assembly and identified an Abd-B ortholog in the scorpionfly Panorpa liui. Quantitative real-time reverse transcription PCR showed that Abd-B expression increased gradually in embryos 76 h post oviposition, and was mainly present in the more posterior abdominal segments. Embryonic RNA interference of Abd-B resulted in a set of abnormalities, including developmental arrest, malformed suckers and misspecification of posterior segment identity. These results suggest that Abd-B is required for the proper development of the posterior abdomen. Furthermore, in Abd-B RNAi embryos, the expression of the appendage marker Distal-less (Dll) was up-regulated and was additionally present on abdominal segments IX and X compared with wild embryos, suggesting that scorpionfly Abd-B may act to suppress proleg development and has gained the ability to repress Dll expression on the more posterior abdominal segments. This study provides additional information on both the functional and evolutionary roles of Abd-B across different insects.

A Matrix Prediction Model for the 6-Month Mortality Risk in Patients With Anti-Melanoma Differentiation-Associated Protein-5-Positive Dermatomyositis.

Objectives: The purpose of this study was to investigate the baseline independent risk factors for predicting 6-month mortality of patients with anti-melanoma differentiation-associated gene 5 (anti-MDA5)-positive dermatomyositis (DM) and develop a matrix prediction model formed by these risk factors. Methods: The hospitalized patients with DM who completed at least 6-month follow-up were recruited as a derivation cohort. The primary exposure was defined as positive anti-MDA5 at the baseline. The primary outcome was all-cause 6-month mortality after enrollment. A matrix prediction model was developed in the derivation cohort, and another published cohort was used for external validation. Results: In derivation cohort, 82 patients with DM were enrolled (mean age of onset 50 ± 11 years and 63% women), with 40 (49%) showing positive anti-MDA5. Gottron sign/papules (OR: 5.135, 95%CI: 1.489-17.708), arthritis (OR: 5.184, 95%CI: 1.455-18.467), interstitial lung disease (OR: 7.034, 95%CI: 1.157-42.785), and higher level of C4 (OR: 1.010, 95%CI: 1.002-1.017) were the independent associators with positive anti-MDA5 in patients with DM. Patients with anti-MDA5-positive DM had significant higher 6-month all-cause mortality than those with anti-MDA5-negative (30 vs. 0%). Among the patients with anti-MDA5-positive DM, compared to the survivors, non-survivors had significantly advanced age of onset (59 ± 6 years vs. 46 ± 9 years), higher rates of fever (75 vs. 18%), positive carcinoma embryonic antigen (CEA, 75 vs. 14%), higher level of ferritin (median 2,858 ug/L vs. 619 ug/L, all p < 0.05). A stepwise multivariate Cox regression showed that ferritin ≥1,250 µg/L (HR: 10.4, 95%CI: 1.8-59.9), fever (HR: 11.2, 95%CI: 2.5-49.9), and positive CEA (HR: 5.2, 95%CI: 1.0-25.7) were the independent risk factors of 6-month mortality. A matrix prediction model was built to stratify patients with anti-MDA5-positive DM into different subgroups with various probabilities of 6-month mortality risk. In an external validation cohort, the observed 6-month all-cause mortality was 78% in high-risk group, 43% in moderate-risk group, and 25% in low-risk group, which shows good accuracy of the model. Conclusion: Baseline characteristics such as fever, ferritin ≥1,250 µg/L, and positive CEA are the independent risk factors for 6-month all-cause mortality in patients with anti-MDA5-positive DM. A novel matrix prediction model composed of these three clinical indicators is first proposed to provide a chance for the exploration of individual treatment strategies in anti-MDA5-positive DM subgroups with various probabilities of mortality risk.

Interaction between Schwann cells and other cells during repair of peripheral nerve injury.

Peripheral nerve injury (PNI) is common and, unlike damage to the central nervous system injured nerves can effectively regenerate depending on the location and severity of injury. Peripheral myelinating glia, Schwann cells (SCs), interact with various cells in and around the injury site and are important for debris elimination, repair, and nerve regeneration. Following PNI, Wallerian degeneration of the distal stump is rapidly initiated by degeneration of damaged axons followed by morphologic changes in SCs and the recruitment of circulating macrophages. Interaction with fibroblasts from the injured nerve microenvironment also plays a role in nerve repair. The replication and migration of injury-induced dedifferentiated SCs are also important in repairing the nerve. In particular, SC migration stimulates axonal regeneration and subsequent myelination of regenerated nerve fibers. This mobility increases SC interactions with other cells in the nerve and the exogenous environment, which influence SC behavior post-injury. Following PNI, SCs directly and indirectly interact with other SCs, fibroblasts, and macrophages. In addition, the inter- and intracellular mechanisms that underlie morphological and functional changes in SCs following PNI still require further research to explain known phenomena and less understood cell-specific roles in the repair of the injured peripheral nerve. This review provides a basic assessment of SC function post-PNI, as well as a more comprehensive evaluation of the literature concerning the SC interactions with macrophages and fibroblasts that can influence SC behavior and, ultimately, repair of the injured nerve.

Assessment of lymphocytic function in vitro stimulated by specific tumor polypeptide combined with dendritic cells / 北京大学学报(医学版)

OBJECTIVE@#To assess the activation function of specific tumor polypeptide for dendritic cell vaccine on lymphocytes proliferation, production of cytokines and killing activity in vitro by using dendritic cells as antigen presenting vector.@*METHODS@#Peripheral blood dendritic cells (DC) and cytokine-induced killer (CIK) were isolated and cultured by adherent culture method; CCK-8 method was used to assess the proliferation function of lymphocytes and the killing function of lymphocytes to tumor cells; enzyme-linked immunospot assay method was used to evaluate the secretion function of cytokines. The experiment was divided into tumor polypeptide group (peptide with DC-CIK), DC-CIK group and CIK group.@*RESULTS@#With presence of interleukin-2 (IL-2) in the culture system, the lymphocyte proliferation of the three groups was obvious. The absorbance at 450 nm of tumor polypeptide group was significantly higher than that of CIK group at the time points day 4 and day 6 (day 4: Z=-3.79, P < 0.001; day 6: Z =-2.95, P < 0.01). The absorbance at 450 nm of group tumor polypeptide was significantly higher than that of DC-CIK group on day 4 (Z=-2.02, P < 0.05). Without IL-2 in the culture system, lymphocytes proliferated slowly in all the three groups, and there was no significant difference in 450 nm absorbance at each time point. The levels of IL-4 (Z=-2.61, P < 0.01), granulocyte-macrophage colony-stimulation factor (GM-CSF, Z=-3.85, P < 0.001), interferon- γ (IFN- γ, Z=-3.56, P < 0.001) and tumor necrosis factor-α (TNF-ɑ, Z=-3.40, P < 0.001) of tumor polypeptide group were higher than those of CIK group. There was no significant difference in the production of cytokines except IL-4 (Z=-2.15, P < 0.05) when tumor polypeptide group was compared with DC-CIK group. The production of IFN-γ (Z=-2.44, P < 0.05), TNF-ɑ (Z=-2.26, P < 0.05) and GM-CSF (Z=-3.73, P < 0.001) in DC-CIK group were higher than those of CIK group. Although there was no significant difference in killing activity between tumor polypeptide group, DC-CIK group and CIK group at hour 18 and hour 24, and the killing activity of tumor polypeptide group was higher than that of the other two groups.@*CONCLUSION@#Tumor peptide combined with dendritic cells can improve the proliferation activity of CIK cells in vitro, and increase the secretion of several cytokines.

Research on the characteristics of calls from psychological assistance hotline and the coping strategies of operators before and after the outbreak of COVID-19 in Changchun City / 四川精神卫生

ObjectiveTo discuss the characteristics of psychological assistance hotline calls and operators' coping strategies of before and after the COVID-19 outbreak， in order to further improve the assistance ability of the psychological crisis hotline. MethodsA retrospective study was conducted on the demographics characteristics， call problems， coping strategies， and call time trends recorded by Changchun psychological assistance hotline information registration platform before the epidemic in Changchun City （January 20， 2019-April 20， 2019） and during the epidemic period （January 20， 2020-April 20， 2020）. ResultsThe differences between gender， age， marital status， location， and occupation type before and during the epidemic were statistically significant （χ2=11.205， 234.240， 152.083， 265.458， 353.385， P<0.01）. The number of different help calls had a statistically significant difference before and during the epidemic （χ2=185.088，P<0.01）. The difference in the number of operators’ different coping strategies before and during the epidemic was statistically significant （χ2=226.810， P<0.01）. Before the epidemic， the main peak of incoming calls was concentrated at 16∶00 to 17∶00， and the secondary peak was concentrated at 22∶00 to 23∶00. During the epidemic， the main peak of incoming calls was also concentrated at 16∶00 to 17∶00， while the secondary peak was concentrated at 10∶00 to 11∶00. ConclusionDuring the COVID-19 epidemic， the number of calls to the psychological assistance hotline was higher than that before the outbreak. The main peak time for calls was the same， and the secondary peak was adjusted from 22∶00 to 23∶00 to 10∶00 to 11∶00. During the epidemic， the number of calls from male， 30 to 39 years old， married， local and staff in Changchun was the most， psychological problems counseling and operator referral strategy were the most before and after the epidemic.

A new phloroglucinol compound from Dryopteris fragrans / 中国中药杂志

Two phloroglucinol compounds(1-2) were isolated and purified from 95% ethanol extract of Dryopteris fragrans through various column chromatographies on silica gel, Sephadex LH-20, medium pressure column chromatography, and preparative HPLC. Their structures were elucidated as 2',4',6'-trihydroxy-5'-methyl acetate-3'-methyl-1'-butyrophenone(1) and aspidinol B(2) based on their chemical and physicochemical methods and spectroscopic data. Compound 1 is a new phloroglucinol compound named "dryofraginol".

Liver with Liver Stagnation and Spleen Deficiency Syndrome and Intestinal Microflora / 中国实验方剂学杂志

Objective:To study the clinical efficacy of Chaihu Shugansan on non-alcoholic fatty liver（NAFLD） patients with liver stagnation and spleen deficiency syndrome and its effect on intestinal flora. Method:The study was a single-center， randomized，single-blind， placebo-controlled clinical study involving 80 patients with NAFLD treated from January 2019 to January 2020 at our hospital. They were divided into two groups （Chaihu Shugansan group，<italic>n</italic>=40） and control group （placebo group，<italic>n</italic>=40）. The two groups of patients were given lifestyle intervention as the basic protocol. The treatment group was orally given Chaihu Shugansan，and the control group was orally given placebo. The drugs were given twice in the morning and evening， 1 dose/time. Patients were followed up for 12 weeks. Before and after treatment，the efficady on liver steatosis was observed by abdominal ultrasound and transient elastography （Fibroscan）， levels of alanine aminotransferase（ALT），aspartate aminotransferase（AST），glutamyl transpeptidase（<italic>γ</italic>-GT），high density lipoprotein cholesterol（HDL-C），low density lipoprotein cholesterol（LDL-C），total cholesterol（TC），triglyceride（TG），interleukin（IL）-6，IL-1<italic>β</italic>，Toll-like receptor-4（TLR-4） in peripheral blood mononuclear cells（PBMCs） and intestinal flora were also detected. Result:There were 37 patients in the treatment group and 35 patients in the control group who finally completed the study protocol. The total effective rate of NAFLD in the treatment group（81.08%，30/37） was higher than that in the control group （68.57%，24/35）（<italic>Z</italic>=2.67，<italic>P</italic><0.05）. The treatment group was superior to the control group in reducing the levels of BMI，ALT，AST，TC，LDL-C，TG，<italic>γ</italic>-GT and increasing the level of HDL-C（<italic>P</italic><0.05）. The levels of pro-inflammatory cytokines（TNF-<italic>α</italic>，IL-1<italic>β</italic> and IL-6），the values of Controlled Attenuation Parameter（CAP），Liver Stiffness Measurement（LSM） and expression of TLR4 were down-regulated in the treatment group （<italic>P</italic><0.01）. In addition，the treatment group showed increase in the abundance of beneficial bacteria （<italic>Bifidobacterium</italic> and <italic>Lactobacillus</italic>） and inhibited the abundance of pathogenic bacteria （<italic>Enterobacter </italic>and<italic> Enterococcus</italic>） in the gut（<italic>P</italic><0.01）. Conclusion:In addition to the lifestyle intervention，Chaihu Shugansan can improve lipid metabolism and liver function，regulate intestinal flora and inhibit the level of inflammatory factors in patients with NAFLD.

CPEB3 regulates neuron-specific alternative splicing and involves neurogenesis gene expression.

In the mammalian brain, alternative pre-mRNA splicing is a fundamental mechanism that modifies neuronal function dynamically where secretion of different splice variants regulates neurogenesis, development, pathfinding, maintenance, migration, and synaptogenesis. Sequence-specific RNA-Binding Protein CPEB3 has distinctive isoform-distinct biochemical interactions and neuronal development assembly roles. Nonetheless, the mechanisms moderating splice isoform options remain unclear. To establish the modulatory trend of CPEB3, we cloned and excessively expressed CPEB3 in HT22 cells. We used RNA-seq to analyze CPEB3-regulated alternative splicing on control and CPEB3-overexpressing cells. Consequently, we used iRIP-seq to identify CPEB-binding targets. We additionally validated CPEB3-modulated genes using RT-qPCR. CPEB3 overexpression had insignificant effects on gene expression in HT22 cells. Notably, CPEB3 partially modulated differential gene splicing enhanced in the modulation of neural development, neuron cycle, neurotrophin, synapse, and specific development pathway, implying an alternative splicing regulatory mechanism associated with neurogenesis. Moreover, qRT-PCR verified the CPEB3-modulated transcription of neurogenesis genes LCN2 and NAV2, synaptogenesis gene CYLD, as well as neural development gene JADE1. Herein, we established that CPEB3 is a critical modulator of alternative splicing in neurogenesis, which remarkably enhances the current understanding of the CPEB3 mediated alternative pre-mRNA splicing.

Ultra-low dose rituximab as add-on therapy in anti-MDA5-positive patients with polymyositis /dermatomyositis associated ILD.

OBJECTIVES: To evaluate the efficacy and safety of ultra-low dose (100 mg) rituximab (RTX) administration in anti-melanoma differentiation-associated gene 5 (MDA5) positive patients with polymyositis/dermatomyositis (PM/DM) associated interstitial lung disease. METHODS: This retrospective study included anti-MDA5 antibody positive ILD subjects in the First Affiliated Hospital of Guangzhou Medical University from November 2017 to March 2019. Independent predictors for 180-day mortality were measured by Cox regression analysis. Patients were divided into 3 groups: Group 1 (non-cyclophosphamide (CTX)/RTX) (n = 10), Group 2 (CTX only) (n = 19) and Group 3 (RTX with/without CTX) (n = 11). The 180-day mortality was compared among 3 groups with Kaplan-Meier analysis. Post-RTX serological parameters as well as adverse events were evaluated. RESULTS: Forty patients were included with the mean age of 51.3 years. Elevated IL-10 level and CD4+/8+ ratio were considered as risk factors of 180-day mortality. Kaplan-Meier analysis showed a trend toward decrease, albeit non-significant, in 180-day mortality in Group 3 (P = 0.26). The administration of 100 mg RTX brought down B cell within 7 days that lasted for 180 days. There were 7 and 6 infection events observed within 2 months of CTX/RTX treatment in Group 2 and 3, with 5 and 2 fatal cases respectively. Cytomegalovirus infection accounted for half infection events in Group 3. CONCLUSION: We found a pronounced and prolonged B cell depletion following 100 mg RTX infusion and RTX add-on may be effective in anti-MDA5 positive ILD patients. However, infection, especially opportunistic infection, should be concerned during the treatment.

Clinical application of autologous freezing fat granule injection grafting in facial rejuvenation / 中华医学美学美容杂志

Objective:To explore the clinical application of autologous freezing fat granule injection grafting in facial rejuvenation.Methods:A total of 64 cases of facial skin soft tissues ageing atrophy were treated by transplantation of autologous purification freezing lipochondria. Autologous fat was obtained from patient＇s abdomen or thighs, centrifugated at low velocity and low pressure to remove the oil and fluid, then stored the lipochondria in -20℃. Rewarming the fat under 37 ℃ for 1 hour, we observed the integrity of the adipocyte and detected the vitality of the fat. Then the purified autologous fat was injected into the recipient site of the face.Results:The fat cell membrane and cell nucleus were clear and integrity after stored in -20℃ for 24 weeks, and the vitality of the fat was (88.89±1.23)%. 21 cases gained satisfactory clinical results by injecting once and 35 cases with 2 times injections, 8 cases with 3 times injections, the effects were satisfactory and there was no complication by follow-up from 6 to 24 months. 82.81% patients and doctors were satisfactory with the curative effect, and 1.56% patients and doctors were unsatisfactory.Conclusions:The effects are satisfactory of autologous purified freezing microparticle fat injected transplant. It has low absorptivity, can duplicate injection, and accept easily by people. It is a good method for facial rejuvenation and worth to spread in the clinical practice.

Advances in studies on chemical constituents, pharmacological effects and clinical application of Aspongopus chinensis / 中国中药杂志

The Aspongopus chinensis is an insect of the genus Hemiptera, which can be used both as a medicinal and as a gourmet in the folk. It has a long history as a drug, which has the effect of regulating Qi and relieving pain, and warming the Yang. It is mainly used to treat stomach cold and pain, liver and stomach pain, kidney deficiency and impotence, and waist and knee pain. Modern pharmacological studies have shown that A. chinensis has a variety of pharmacological activities. For example, it can be used to fight tumors, improve reproductive damage, and antibacterial, anti-oxidation, anti-coagulation, anti-ulcer, anti-fatigue and so on. The chemical constituents of A. chinensis currently reported mainly include odorous components, vitamins, fatty acids and proteins, amino acids, and other nutrients, as well as nucleosides and dopamines. This study summarizes and analyzes the related research literatures of A. chinensis in China and abroad, and provides a reference for its further development and research from the aspects of chemical composition, pharmacological action and clinical application.

Spermiogenesis of the hangingfly Terrobittacus implicatus (Huang and Hua) (Mecoptera: Bittacidae).

The structure of spermatozoa is able to provide valuable characters in resolving phylogenic relationships in Metazoa, especially in insects. Such data, however, are greatly deficient in Mecoptera. Here, we studied the spermiogenesis and ultrastructure of sperm in the hangingfly Terrobittacus implicatus (Huang and Hua) using transmission electron microscopy. The results show that the spermatogenesis of T. implicatus occurs within sperm cysts, following a pattern commonly found in insects. The microtubular doublets of spermatid axoneme exhibit a hooklike projection from the B-subtubule in the early period, but the projection disappears in the mature stage. The mature spermatozoon of T. implicatus is a filiform cell that is pronouncedly elongated and has a bi-layered acrosome, a nucleus with two lateral longitudinal grooves, a neck region with the centriole adjunct, a flagellum with a simple 9 + 2 axoneme, two extra-axonemal accessory structures, two accessory bodies, and two mitochondrial derivatives of unequal size, and a prominent glycocalyx. The basic structure of spermatozoa of T. implicatus is similar to that of other Mecoptera studied. However, this species shows characteristics unique in Bittacidae, such as the reniform appearance of the centriole adjunct, two triangular accessory bodies with granular materials, and two asymmetric mitochondrial derivatives with a circular profile in cross-section. The potential utilization of the sperm ultrastructure for understanding the phylogeny of Bittacidae is briefly discussed.

SAMD9 is a (epi-) genetically regulated anti-inflammatory factor activated in RA patients.

To identify PBMC-expressed genes significant for RA, and to ascertain their upstream regulatory factors, as well as downstream functional effects relevant to RA pathogenesis. We performed peripheral blood mononuclear cells (PBMCs) transcriptome-wide mRNA expression profiling in a case-control discovery sample. Differentially expressed genes (DEGs) were identified and validated in PBMCs in independent samples. We also generated genome-wide SNP genotyping data, and collected miRNA expression data and DNA methylation data from PBMCs of the discovery sample. Pearson correlation analyses were conducted to identify miRNAs/DNA methylations influencing DEG expression. Association analyses were conducted to identify expression-regulating SNPs. The key DEG, SAMD9, which was reported to function as a tumor suppressor gene, was assessed for its effects on T cell proliferation, apoptosis, and inflammatory cytokine expression. A total of 181 DEGs (Fold Change ≥ 2.0, Bonferroni adjusted p ≤ 0.05) were discovered in PBMCs. Four DEGs (SAMD9, CKLF, PARP9, and GUSB), upregulated with RA, were validated independently in PBMCs. Specifically, SAMD9 mRNA expression level was significantly upregulated in PHA-activated Jurkat T cells in vitro, and correlated with 8 miRNAs and associated with 22 SNPs in PBMCs in vivo. Knockdown of SAMD9 could transiently promote Jurkat T cell proliferation within 48 h and significantly induce TNF-α and IL-8 expression in T cells. SAMD9 expression is (epi-) genetically regulated, and significantly upregulated in PBMCs in RA patients and in activated T cells in vitro. SAMD9 might serve as a T cell activation marker but act as an anti-inflammatory factor.

Detection of lncRNA-mRNA interaction modules by integrating eQTL with weighted gene co-expression network analysis.

One major function of lncRNA is to regulate the expression of mRNA, but the patterns of their interactions were largely unknown. We attempted to construct lncRNA-mRNA interaction modules at a genome-wide scale. We performed a genome-wide lncRNA-mRNA eQTL analysis in peripheral blood mononuclear cells of 43 individuals, followed by weighted gene co-expression network analysis and functional enrichment analysis which sought to detect functional modules. There were 4627 significant cis lnc-eQTL pairs (P < 1.4 × 10-6) and 1,587,128 significant trans lnc-eQTL pairs (P < 3.46 × 10-9). We detected 11 eQTL modules for the lnc-eQTL networks. Among them, five modules showed significant enrichments in GO terms, and three modules showed significant enrichments in specific KEGG pathways (e.g., Toll-like receptor, PI3K-Akt, NF-kappa B, and TNF signaling pathways). lncRNA-protein interaction analysis showed that some well-known functional lncRNAs (HOTAIR, CCDC26, RHPN1-AS1, WT1-AS, and TCL6) in the eQTL module interacted with genes in focal adhesion and PI3K-Akt signaling pathway. We identified biologically functional lncRNA-mRNA interaction modules by integrating eQTL and weighted gene co-expression network analysis. Integrative analysis of lncRNA and mRNA data by applying eQTL analysis and weighted gene co-expression network analysis methods could be helpful for functional annotation of lncRNAs.

Analysis of Effective Substance of Qingre Chushi Decoction by UPLC-Q-TOF-MS / 中国实验方剂学杂志

Objective:To characterize and compare the chemical information of four extracts of Qingre Chushi (QRCS) decoction by liquid chromatography-mass spectrometry (LC-MS), and combine the chemical information of the four extracts with their results of anti-inflammatory effect for a multivariate statistical analysis, in order to identify the compounds directly relating to the anti-inflammatory effects of QRCS decoction. Method:Four extracts of QRCS decoction were characterized by UPLC-Q-TOF-MS:①ethanol extract+water extract,② ethanol extract+supernatant after water extraction and alcohol precipitation, ③ ethanol extract+precipitation after water extraction and alcohol precipitation,and ④ standard decoction. On the basis of the results of inhibition of the four above extracts on xylene-induced ear swelling in mice,multivariate statistical analysis[principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA)] were carried out to lock the chromatographic peaks with significant differences between group ① (the best pharmacological action group) and group ④ (standard decoction group). According to the accuracy of quasi-molecular ion and fragment ion data,and the reference materials and literature data,those chromatographic peaks were identified. Result:PCA could cluster the four extracts of QRCS decoction,and the differences between groups was reflected in the distance between groups. Group ④ (standard decoction) had the most significant differences with the other three groups, especially in the first principal component; group ① (ethanol extract+water extract),group ② (ethanol extract+supernatant after water extraction and ethanol precipitation) and group ③ (ethanol extract+precipitation after water extraction and ethanol precipitation) had certain differences in the second principal component. OPLS-DA was used to compare group ① (the best pharmacological action group) and group ④ (standard decoction group). Eleven chromatographic peaks with great contribution and high reliability to group differences,were identified as gentiopicrin,skimmin,baicalin,baicalin isomer,wogonoside,5,6,7-trihydroxy-8-methoxyflavone-7-O-glucurodonaldehyde,5,6-dihydroxy-6,8,2',3'-tetramethoxyflavone,salicin-6-C-arabinose-8-C-glucoside,plantamajoside and glycyrrhizic acid. Conclusion:In the mode of pectrum-effect combination, this study explores and identifies compounds relating to the anti-inflammatory effect of QRCS decoction,so as to provide the basis for screening the extraction and purification process and optimizing the formulation of preparation of Qingre Chushi decoction.

Correlation analyses revealed global microRNA-mRNA expression associations in human peripheral blood mononuclear cells.

MicroRNAs (miRNAs) can regulate gene expression through binding to complementary sites in the 3'-untranslated regions of target mRNAs, which will lead to existence of correlation in expression between miRNA and mRNA. However, the miRNA-mRNA correlation patterns are complex and remain largely unclear yet. To establish the global correlation patterns in human peripheral blood mononuclear cells (PBMCs), multiple miRNA-mRNA correlation analyses and expression quantitative trait locus (eQTL) analysis were conducted in this study. We predicted and achieved 861 miRNA-mRNA pairs (65 miRNAs, 412 mRNAs) using multiple bioinformatics programs, and found global negative miRNA-mRNA correlations in PBMC from all 46 study subjects. Among the 861 pairs of correlations, 19.5% were significant (P < 0.05) and ~70% were negative. The correlation network was complex and highlighted key miRNAs/genes in PBMC. Some miRNAs, such as hsa-miR-29a, hsa-miR-148a, regulate a cluster of target genes. Some genes, e.g., TNRC6A, are regulated by multiple miRNAs. The identified genes tend to be enriched in molecular functions of DNA and RNA binding, and biological processes such as protein transport, regulation of translation and chromatin modification. The results provided a global view of the miRNA-mRNA expression correlation profile in human PBMCs, which would facilitate in-depth investigation of biological functions of key miRNAs/mRNAs and better understanding of the pathogenesis underlying PBMC-related diseases.

Identification of expression quantitative trait loci (eQTLs) in human peripheral blood mononuclear cells (PBMCs) and shared with liver and brain.

PBMCs are essential for immunity and involved in various diseases. To identify genetic variations contributing to PBMCs transcriptome-wide gene expression, we performed a genome-wide eQTL analysis by using genome-wide SNPs data and transcriptome-wide mRNA expression data. To assess whether there are common regulation patterns shared among different tissues/organs, public datasets were utilized to identify common eQTLs shared with PBMCs in lymphoblastoid, monocytes, liver, and brain. Allelic expression imbalance (AEI) assay was employed to validate representative eQTLs identified. We identified 443 cis- and 2386 trans-eSNPs (FDR <0.05), which regulated 128 and 635 target genes, respectively. A transcriptome-wide expression regulation network was constructed, highlighting the importance of 28 pleiotropic eSNPs and 18 dually (cis- and trans-) regulated genes. Three genes, that is, TIPRL, HSPB8, and EGLN3, were commonly regulated by hundreds of eSNPs and constituted a very complex interaction network. Strikingly, the missense SNP rs371513 trans- regulated 25 target genes, which were functionally related to poly(A) RNA binding. Among 8904 eQTLs (P < 0.001) identified herein in PBMCs, a minority (163) was overlapped with lymphoblastoid, monocytes, liver, and/or brain. Besides, two cis-eSNPs in PBMC were confirmed by AEI. The present results demonstrated a comprehensive expression regulation network for human PBMCs and may provide novel insights into the pathogenesis of immunological diseases related to PBMCs.

Multiple correlation analyses revealed complex relationship between DNA methylation and mRNA expression in human peripheral blood mononuclear cells.

DNA methylation is an important regulator on the mRNA expression. However, a genome-wide correlation pattern between DNA methylation and mRNA expression in human peripheral blood mononuclear cells (PBMCs) is largely unknown. The comprehensive relationship between mRNA and DNA methylation was explored by using four types of correlation analyses and a genome-wide methylation-mRNA expression quantitative trait locus (eQTL) analysis in PBMCs in 46 unrelated female subjects. An enrichment analysis was performed to detect biological function for the detected genes. Single pair correlation coefficient (r T1) between methylation level and mRNA is moderate (-0.63-0.62) in intensity, and the negative and positive correlations are nearly equal in quantity. Correlation analysis on each gene (T4) found 60.1% genes showed correlations between mRNA and gene-based methylation at P < 0.05 and more than 5.96% genes presented very strong correlation (R T4 > 0.8). Methylation sites have regulation effects on mRNA expression in eQTL analysis, with more often observations in region of transcription start site (TSS). The genes under significant methylation regulation both in correlation analysis and eQTL analysis tend to cluster to the categories (e.g., transcription, translation, regulation of transcription) that are essential for maintaining the basic life activities of cells. Our findings indicated that DNA methylation has predictive regulation effect on mRNA with a very complex pattern in PBMCs. The results increased our understanding on correlation of methylation and mRNA and also provided useful clues for future epigenetic studies in exploring biological and disease-related regulatory mechanisms in PBMC.