Pro-angiogenic effect of exosomal microRNA-103a in mice with rheumatoid arthritis via the downregulation of hepatocyte nuclear factor 4 alpha and activation of the JAK/STAT3 signaling pathway.

Rheumatoid arthritis (RA) is characterized by inflammation of the synovial membrane, accompanied by hyperplasia and neo-angiogenesis, which promote local inflammation. Macrophage-derived exosomes have been reported to enhance inflammation and the immune response. In the present study, we identified a novel exosomal microRNA (miR)-103a, which aids in the regulation of inflammation and angiogenesis in mice with RA, and attempted to identify the underlying mechanism. Initially, a mouse model of RA was established. Thereafter, exosomes were isolated from macrophage RAW264.7 cells and evaluated through transmission electron microscopy and nanoparticle tracking analysis. After prediction and verification of the target genes of miR-103a, RT-qPCR was used to assess miR-103a and HNF4A expression in mice with RA. High expression of miR-103a and low expression of HNF4A were observed in mice with RA, thus, miR-103a was found to target and downregulate HNF4A. Exosomal miR-103a promoted inflammation and angiogenesis in mice with RA which was accompanied by an increase in the levels of factors associated with inflammation and angiogenesis. However, an opposite trend was observed upon HNF4A elevation. Exosomal miR-103a was also found to activate the JAK/STAT3 signaling pathway. In conclusion, exosomal miR-103a inhibited the expression of HNF4A to activate the JAK/STAT3 signaling pathway, thereby exacerbating RA in mice.

[Application value of metagenomic next-generation sequencing (mNGS) in the diagnosis of different types of tuberculosis].

Objective: To investigate the diagnostic value of metagenomic next generation sequencing(mNGS)for different types of tuberculosis(TB). Methods: A retrospective analysis was performed on suspected TB patients admitted to Shanghai Pulmonary Hospital from January 1, 2019 to December 31, 2019. The patients underwent mNGS test, acid-fast staining smear, Mycobacterium tuberculosis (MTB) culture and Xpert MTB/RIF(Xpert). A total of 205 TB suspects were finally included, including 120 males and 85 females, with a median age of 34 (16, 70) years. There were 165 cases of TB, including 92 males and 73 females, with a median age of 30 (12, 67) years. There were 40 non-TB patients, 28 males and 12 females, with a median age of 44.6 (24, 78) years, including 28 cases of infectious diseases and 12 cases of malignant tumors. With the final diagnosis as the reference standard, the diagnostic efficiency of mNGS in different types of tuberculosis (including pulmonary tuberculosis and extrapulmonary tuberculosis) and differences in diagnostic sensitivities between mNGS and other detection methods were calculated, respectively. P<0.05 was considered to be statistically different. Results: With clinical diagnosis as the reference standard, the sensitivity of mNGS to diagnose TB in this study was 60% (99/165, 95%CI: 0.53-0.67), and the specificity was 100% (40/40, 95%CI 0.90-1.00). The positive predictive value was 100% (99/99, 95%CI: 0.95-1.00), and the negative predictive value was 37.74% (40/106, 95%CI: 0.29-0.48). The sensitivity of mNGS in TB was 60% (99/165), higher than that of smear (10.9%, 18/165), Xpert(43.64%, 72/165) and culture (23.03%, 38/165). All the differences were statistically significant (P=0.00). The sensitivity of mNGS in pulmonary TB was 59%(59/100), which was significantly higher than that of smear (15%, 15/100) and culture (26%, 26/100), all P<0.001, but the difference was not statistically significant compared with Xpert (52%,52/100, P=0.09). In the extrapulmonary tuberculosis (EPTB), the sensitivity of mNGS was 61.54% (40/65), higher than that of acid fast staining(4.62%,3/65), Xpert (30.77%, 20/65)and culture (18.45%, 12/65). All the differences were statistically significant (all P<0.001). Conclusion: The mNGS was rapid and efficient in detection of MTB, and was superior to other traditional diagnostic methods for TB, especially for EPTB.

[Impact of the COVID-19 epidemic on anxiety among the elderly in community].

Objective: To compare the prevalence of anxiety among old people before and during the COVID-19 epidemic in China, and to provide scientific evidence for psychological intervention of the elderly during public health emergencies. Methods: In 2019, the National Psychological Care Project for the Elderly was launched, covering 818 communities across the country, and 188 407 subjects received psychological assessment. In April and May 2020, a convenient sample of 6 467 aged 65 and above subjects were followed up on the anxiety status and its influencing factors during the epidemic period by using structurized questionnaire. Data collection and management were carried out using the national elderly psychological care project data collection platform. McNemar test was used to compare the difference of the prevalence of anxiety among elderly before (October 2019 to January 23, 2020) and during the epidemic (April-May 2020). The difference of the prevalence of anxiety among elderly with different characteristics was compared by chi square test. The influencing factors of anxiety before and during the epidemic situation were analyzed by multivariate logistic regression model. Results: The prevalence of anxiety symptoms in the elderly population was 4.95% (95%CI: 4.42%-5.48%) before the outbreak of COVID-19, and 10.10% (95%CI: 9.36%-10.83%) during the epidemic which was twice as high as before the outbreak. The difference was statistically significant (P<0.05). Multivariate logistic regression analysis showed that the risk factors of anxiety symptoms before the outbreak were with one underlying disease (OR=1.57, 95%CI: 1.05-2.37), with two or more underlying diseases (OR=3.10, 95%CI: 2.13-4.51), and the protective factors were with hobbies, good relationship between children, good relationship with spouse, positive aging attitude and good psychological resilience (all P<0.05). The risk factors of developing anxiety symptoms during the epidemic were living in rural areas (OR=1.77, 95%CI: 1.42-2.20), participating in social activities regularly (OR=1.23, 95%CI: 1.02-1.48), having a good relationship with friends (OR=1.42, 95%CI: 1.11-1.82) and were quarantined or people around were quarantined for medical observation (OR=2.80, 95% CI: 1.90-4.13). Conclusion: The COVID-19 epidemic leads to a double increase in anxiety among the elderly. We should pay more attention to the psychological state of the elderly in rural area and who is being quarantined or people around being quarantined for medical observation.

[Correlation between elevation of brachial artery pulse pressure increased and coronary heart disease in different genders].

Objective: To explore the relationship between elevation of brachial pulse pressure and coronary heart disease in different genders. Methods: A total of 5 116 inpatients with suspected stable coronary heart disease were consecutively enrolled from December 2011 to June 2017 in the Affiliated Hospital of Chengde Medical College, and divided into coronary heart disease group (n=3 694) and non-coronary heart disease group (n=1 422). The clinical data of all inpatients were collected. A binary logistic regression model of coronary heart disease in different genders were separately established. Results: The morbidity of hypertension, dyslipidemia, type 2 diabetes, ischemic stroke and elevated pulse pressure were all higher in the coronary heart disease group than those in the non-coronary heart disease group (all P<0.05). The area under curve (AUC) of pulse pressure in the male group was 0.540, with an optimal diagnostic threshold of 50 mmHg. The AUC of pulse pressure in the female group was 0.612, with an optimal diagnostic threshold of 60 mmHg. Besides hypertension, type 2 diabetes, increase of low-density lipoprotein cholesterol, ischemic stroke, smoking, and aging, elevated pulse pressure was also an independent risk factor for coronary heart disease in both male and female groups (all P<0.05). Additionally, the risk of elevated pulse pressure for coronary heart disease was higher in female group than that of male group (odds ratio (OR): 1.741 vs 1.284, P<0.05). Conclusion: Elevated pulse pressure may be a new risk factor for coronary heart disease, and its risk for coronary heart disease is higher in women than in men.

[Gastric adenocarcinoma of the fundic gland type: clinicopathological analysis of six cases].

Objective: To investigate the clinicopathological characteristics of gastric adenocarcinoma of the fundic gland(GA-FG). Methods: A total of 6 patients with GA-FG admitted from January 2014 to May 2019 were identified at Shanghai Songjiang District Central Hospital.Analyzed endoscopic findings, pathological characteristics and immunohistochemical staining of the lesions and reviewed relevant literatures. Results: Among the 6 patients, 3 were male, 3 were female, and the mean age was 66 years. The most common clinical presentation was epigastric discomfort. Gastroscopic examination revealed an elevated or flat lesion in the body of the stomach, with a diameter of 0.4 to 1.0 cm. Histopathologically, they mainly composed of main cells, but there were also scattered parietal cells, arranged in an irregular tubular or cord-like structure, and formed anastomosis branches. The tumor cells were immunohistochemically positive for MUC6 and pepsinogen 1. Conclusions: GA-FG is a rare variant of well-differentiated adenocarcinoma. It is important to understand its clinical features and histological morphology for more accurate diagnosis.

[Epidemiological characteristics of COVID-19 family clustering in Zhejiang Province].

Objective: To investigate the epidemiological characteristics for family clusters of COVID-19 in Zhejiang Province. Methods: The data including cases information of asymptomatic infected cases of family clusters of COVID-19 in Zhejiang Province were collected from Public Health Emergencies Reporting System of China Center for Disease Control and Prevention. Calculate the case number of subsequent cases, index cases, asymptomatic infected cases, exposure cases, and then, compute family secondary attack rate (SIR) and serial interval. Results: A total of 389 cases comprised 149 family index cases and 240 subsequent cases. The clinical symptoms between family index cases and subsequent cases (exclude asymptomatic infected cases ) were similar, fever was the most common symptoms in the two groups 115 (77.18%) and 110（48.67%）respectively, the cases with diarrhea symptoms accounted for the least proportion, which were 7 (4.70%) and 6 (2.65%) respectively. The serial interval between the family index cases and the subsequent cases [M (P25, P75)] was 4.00 (2.00, 6.00) days. Family secondary attack rate for subsequent cases was 34.43%, subsequent cases aged between 14 and 60 have the highest SIR (43.42%) compared with other two age groups, the difference was statistically (P<0.001); the family SIR of the spouses of the family index cases is 68.57%, and are higher than that of parents (29.03%), children (25.00%) and other family members (24.21%), the difference was also statistically (P<0.001). Conclusion: 2019 novel coronavirus has shorter serial interval and higher family SIR, the SIR of spouses is higher than other family members.

Effects of sodium lactate Ringer's injection on transfection of human protein kinase C-α antisense oligonucleotide in A549 lung cancer cells.

In the present study, we evaluated the effects of four solutions [Dulbecco's modified Eagle's medium (DMEM), sodium lactate Ringer's injection (SLRI), phosphate-buffered saline (PBS), and NaCl] on the transfection of the human protein kinase C-a antisense oligonucleotide (PKC-a ASO) aprinocarsen in human lung carcinoma A549 cells. Specifically, SLRI, DMEM, PBS, or NaCl were used as the growth solutions for A549 cells, and OPTI-MEM was used as the PKC-a ASO diluent for transfection. Additionally, SLRI, DMEM, PBS, or NaCl were used as both the growth solutions and diluents for transfection. The cell viability and transfection efficiency were determined. The results demonstrated that when SLRI was used as either the growth solution or both the growth solution and diluent for aprinocarsen transfection in A549 cells, the effects were close to the best effects observed with DMEM as the growth solution and OPTI-MEM as the diluent, which supported the transfection of aprinocarsen into the cells. Moreover, SLRI resulted in higher transfection efficiency than those of PBS and NaCl. In in vitro experiments, aprinocarsen effectively induced apoptosis in A549 cells. In conclusion, SLRI may replace PBS or NaCl in clinical trials as a transfection solution readily accepted by the human body. To our knowledge, this is the first report demonstrating the use of SLRI as a transfection solution in lung-cancer cell lines.

[Impacts of airborne particulate matter and its components on respiratory system health].

Nowadays, particulate air pollution has been a global environmental problem. Numerous studies has shown that long-term exposure to high level of airborne particulate matter (PM) can damage human health. Respiratory system, as a direct portal to contact with particulate matter, can be more susceptible to airborne particulates. Summarizing latest five-year epidemiological research, the present review is focused on the effects of PM on respiratory system health in different age groups. In detail, we investigated the harmful effect of PM, or its components on three common respiratory diseases, including lung function decline, chronic obstructive pulmonary disease (COPD) and asthma. The result showed that, to a certain degree, PM could induce the decline of lung function, the development and the exacerbation of COPD and asthma by oxidative stress and inflammatory reaction. And it may prompt that exposure to PM can be an improtant risk factor for the respiratory system health.

Electrophysiological Differences between the Same Pore Region Mutation in SCN1A and SCN3A.

Mutations in the sodium channel gene, SCN1A (NaV1.1), have been linked to a spectrum of epilepsy syndromes, and many of these mutations occur in the pore region of the channel. Electrophysiological characterization has revealed that most SCN1A mutations in the pore region result in complete loss of function. SCN3A mutations have also been identified in patients with epilepsy; however, mutations in this pore region maintain some degree of electrophysiological function. It is thus speculated that compared to SCN3A disruptions, SCN1A mutations have a more pronounced effect on electrophysiological function. In this study, we identified a novel mutation, N302S, in the SCN3A pore region of a child with epilepsy. To investigate if mutations at the pore regions of SCN3A and SCN1A have different impacts on channel function, we studied the electrophysiological properties of N302S in NaV1.3 and its homologous mutation (with the same amino acid substitution) in NaV1.1 (N301S). Functional analysis demonstrated that SCN1A-N301S had no measurable sodium current, indicating a complete loss of function, while SCN3A-N302S slightly reduced channel activity. This observation indicates that the same pore region mutation affects SCN1A more than SCN3A. Our study further revealed a huge difference in electrophysiological function between SCN1A and SCN3A mutations in the pore region; this might explain the more common SCN1A mutations detected in patients with epilepsy and the more severe phenotypes associated with these mutations.

miR-506 acts as a tumor suppressor by directly targeting the hedgehog pathway transcription factor Gli3 in human cervical cancer.

Although significant advances have recently been made in the diagnosis and treatment of cervical carcinoma, the long-term survival rate for advanced cervical cancer remains low. Therefore, an urgent need exists to both uncover the molecular mechanisms and identify potential therapeutic targets for the treatment of cervical cancer. MicroRNAs (miRNAs) have important roles in cancer progression and could be used as either potential therapeutic agents or targets. miR-506 is a component of an X chromosome-linked miRNA cluster. The biological functions of miR-506 have not been well established. In this study, we found that miR-506 expression was downregulated in approximately 80% of the cervical cancer samples examined and inversely correlated with the expression of Ki-67, a marker of cell proliferation. Gain-of-function and loss-of-function studies in human cervical cancer, Caski and SiHa cells, demonstrated that miR-506 acts as a tumor suppressor by inhibiting cervical cancer growth in vitro and in vivo. Further studies showed that miR-506 induced cell cycle arrest at the G1/S transition, and enhanced apoptosis and chemosensitivity of cervical cancer cell. We subsequently identified Gli3, a hedgehog pathway transcription factor, as a direct target of miR-506 in cervical cancer. Furthermore, Gli3 silencing recapitulated the effects of miR-506, and reintroduction of Gli3 abrogated miR-506-induced cell growth arrest and apoptosis. Taken together, we conclude that miR-506 exerts its anti-proliferative function by directly targeting Gli3. This newly identified miR-506/Gli3 axis provides further insight into the pathogenesis of cervical cancer and indicates a potential novel therapeutic agent for the treatment of cervical cancer.

Hydrogen peroxide enhances the uptake of polyethylenimine/oligonucleotide complexes in A549 cells by activating CaMKII independent of [Ca(2+)]c elevation.

Aerosol oligonucleotide therapy has vast potential in pulmonary system diseases. Reactive oxygen species (ROS) play an important role in complex physiological processes such as cell signaling, apoptosis, etc. Therefore, to determine the mechanism of ROS involvement in polyethylenimine/oligonucleotide (PEI/ON) endocytosis in cells, we measured the fluorescence intensities of fluorescein isothiocyanate-labeled ON complexes with PEI and the changes in cytosolic Ca(2+) concentration ([Ca(2+)]c) in A549 cells after hydrogen peroxide (H2O2) stimulation. Results showed that improved uptake of PEI/ON complexes was independent of the rise of [Ca(2+)]c in A549 cells, including the Ca(2+) inflow and the release of Ca(2+) from intracellular stores induced by 500 µM H2O2. However, the enhanced uptake efficiency was almost completely abolished by the calcium/calmodulin-dependent protein kinase II (CaMKII) inhibitor and the microtube depolymerized drug. CaMKII-dependent microtube polymerization may be responsible for the enhanced uptake of PEI/ON complexes in A549 cells under oxidative stress conditions. This study is useful for research aimed at improving aerosol oligonucleotide therapy in pulmonary system diseases.

AtMYB44 regulates resistance to the green peach aphid and diamondback moth by activating EIN2-affected defences in Arabidopsis.

Recently we showed that the transcription activator AtMYB44 regulates expression of EIN2, a gene essential for ethylene signalling and insect resistance, in Arabidopsis thaliana (Arabidopsis). To link the transactivation with insect resistance, we investigated the wild-type and atmyb44 mutant plants, genetically Complemented atmyb44 (Catmyb44) and AtMYB44-Overexpression Transgenic Arabidopsis (MYB44OTA). We found that AtMYB44 played a critical role in Arabidopsis resistance to the phloem-feeding generalist green peach aphid (Myzus persicae Sulzer) and leaf-chewing specialist caterpillar diamondback moth (Plutella xylostella L.). AtMYB44 was required not only for the development of constitutive resistance but also for the induction of resistance by both herbivorous insects. Levels of constitutive and herbivore-induced resistance were consistent with corresponding amounts of the AtMYB44 protein constitutively produced in MYB44OTA and induced by herbivory in Catmyb44. In both cases, AtMYB44 promoted EIN2 expression to a greater extent in MYB44OTA than in Catmyb44. However, AtMYB44-promoted EIN2 expression was arrested with reduced resistance levels in the EIN2-deficient Arabidopsis mutant ein2-1 and the MYB44OTA ein2-1 hybrid. In the different plant genotypes, only MYB44OTA constitutively displayed phloem-based defences, which are specific to phloem-feeding insects, and robust expression of genes involved in the biosynthesis of glucosinolates, which are the secondary plant metabolites known as deterrents to generalist herbivores. Phloem-based defences and glucosinolate-related gene expression were not detected in ein2-1 and MYB44OTA ein2-1. These results establish a genetic connection between the regulatory role of AtMYB44 in EIN2 expression and the development of Arabidopsis resistance to insects.

Secreted factor R-Spondin 2 is involved in refinement of patterning of the mammalian cochlea.

BACKGROUND: In the cochlea, patterning of the organ of Corti is tightly regulated to produce a single row of sound-detecting inner hair cells and three rows of outer hair cells, which amplify and refine the signal. The recently identified R-Spondin family of signaling molecules usually act as co-activators of Wnt signaling; it is thought that they regulate turnover of Wnt receptors at the membrane. We sought to test whether R-Spondins function in the developing cochlea. RESULTS: Expression analysis of all four members of the R-Spondin family showed that only R-Spondin2 (Rspo2) is expressed in the cochlea during development of the sensory epithelium. Examination of an Rspo2(-/-) mouse showed that loss of Rspo2 results in an additional single row of outer hair cells and disruption of peripheral innervation pattern. Addition of Rspo2 recombinant protein to organotypic cochlear cultures resulted in a small but significant decrease in the number of outer hair cells. CONCLUSIONS: Rspo2 is required to limit the number of outer hair cells to three rows and for optimal arrangement of peripheral nerve fibers. The Rspo2 gain- and loss-of-function studies show that in the ear, Rspo2 function is not consistent with its assigned role as a Wnt potentiator.

Altered expression of voltage-gated potassium channel 4.2 and voltage-gated potassium channel 4-interacting protein, and changes in intracellular calcium levels following lithium-pilocarpine-induced status epilepticus.

The A-type voltage-gated potassium channels (Kv4) have been proved to play a major role as modulators of somatodendritic excitability. Recent studies indicate that neuronal hyperactivity in epilepsy is associated with changes in Kv4. However, the precise regulation of Kv4 in the development of epilepsy and its underlying mechanism remain unclear. In this study, we investigated whether the expression of the Kv4.2 channel and of its major modulator, voltage-dependent potassium channel-interacting protein (KChIP1), is altered following lithium-pilocarpine induced status epilepticus (SE) and the chronic-epilepsy phase in the rat model. We found that Kv4.2 and KChIP1 expression was transiently up-regulated following SE, whereas it was down-regulated during the chronic phase: this was most prominent in the CA1 and CA3 regions. The time-course analysis of the protein expression level showed that the peak Kv4.2 up-regulation was between 6 and 24 h after SE, whereas KChIP1 expression was increased earlier and for a shorter period. The temporospatial changes in Kv4.2 were very similar to those of its major modulator KChIP1. We compared the difference in 4-aminopyridine (4-AP)-induced intracellular calcium ([Ca(2+)]i) elevation between model and control brain slices. The results showed that the [Ca(2+)]i elevation induced by the Kv4 channel blocker 4-AP was aggravated and prolonged in the model slice after SE. The functional relevance of these changes in Ca(2+) homeostasis and Kv4.2 and KChIP1 expression may be associated with intrinsic neuronal excitability regulation and epileptogenesis.

A family-based and case-control association study of the NOTCH4 gene and schizophrenia.

Recently a strong positive association between schizophrenia and Notch4 has been reported. Both individual markers and haplotypes showed association with the disease, with five markers (three microsatellites and two SNPs) being tested. In order to test this finding we genotyped these markers in the Han Chinese population using a sample of 544 cases and 621 controls as well as >300 trios. Analysis of allele, genotype and haplotype frequencies in both samples showed no association between the markers and the disease. Our results would indicate that a significant role for the Notch4 gene in schizophrenia can be ruled out in the Han Chinese. However, similar studies are necessary in the Caucasian population as linkage disequilibrium arrangements and founder effects may differ between these two populations.

Binding characteristics of M and L isoantibodies to high and low potassium sheep red cells.

Binding of highly purified 125I labeled M and L antibodies, both belonging to the immunoglobulin G class, was studied in high potassium (HK) and low potassium (LK) sheep red cells. Anti-M and anti-L bound specifically to M and L antigen positive HK and LK red cells, respectively. Nonspecific binding was higher for anti-L to HK cells than for anti-M to LK cells. Once bound, the M and L antibodies were capable of inducing complement dependent immune hemolysis. Only 75-100 and 500-750 molecules of anti-M and anti-L immunoglobulins were required to hemolyze 50% of HK (MM) and LK (LL) red cells, respectively, suggesting that the M and L antigens may be clustered on the surfaces of these cells. Equilibrium binding studies revealed that the maximum number of M sites is 3-6 x 10(3) in HK (MM) and 1.5-4 x 10(3) in LK (LM) cells, respectively. In comparison, the number of L antigens is slightly lower in LK cells, about 1.2-1.8 x 10(3) in LL and less in LM(LK) red cells. The number of M and L antigens, therefore, is more than an order of magnitude larger than that of the Na+K+ pumps measured previously in these cells by 3H-ouabain binding, thus precluding a quantitative correlation between M and L antigens and the Na+K+ pumps different in the three genetic types of sheep red cells. The binding affinities of both anti-M and anti-L could not be described by a single equilibrium dissociation constant indicating heterogeneous antibody populations and /or variability in the antigenic sets of individual HK or LK cells. The pronounced heterogeneity of antigens and/or antibodies in both the M and L systems was reflected in the antibody association kinetics, which also exhibited a remarkable temperature dependence. The data suggest that the correlation between the M and L antigens and the Na+K+ pump molecules is more complex than that in goat red cells previously reported by others.