A preliminary study on the normal value of the thoracic haller index in children.

BACKGROUND AND OBJECTIVE: The Haller index (HI) is widely utilized as a quantitative indicator to assess the extent of pectus excavatum (PE) deformity, which is the most common chest wall abnormality in children. Both preoperative correction planning and postoperative follow-up need to be based on the standard of normal thoracic growth and development. However, there is currently no established reference range for the HI in children. Consequently, the aim of this study was to conduct a preliminary investigation of normal HI values among children to understand thoracic developmental characteristics. METHODS: Chest CT images obtained from January 2012 to March 2022 were randomly selected from the imaging system of the Children's Hospital of Chongqing Medical University. We divided the images of children into a total of 19 groups: aged 0-3 months (1 group), 4-12 months (1 group), and 1 year to 17 years (17 groups), with 50 males and 50 females, totaling 100 children in each group. HI was measured in the plane where the lowest point of the anterior thoracic wall was located and statistically analyzed using SPSS 26.0 software. RESULTS: A total of 1900 patients were included in the study. Our results showed that HI, transverse diameter, and anterior-posterior diameter were positively correlated with age (P < 0.05). Using age as the independent variable and HI as the dependent variable, the best-fit regression equations were HI-male = 2.047 * Age0.054(R2 = 0.276, P＜0.0001), HI-female = 2.045 * Age0.067(R2 = 0.398, P＜0.0001). Males had significantly larger thoracic diameters than females, and there was little difference in the HI between the two sexes. CONCLUSIONS: The HI rapidly increases during the neonatal period, slowly increases during infancy, and stops increasing during puberty, with no significant differences between sexes.

Aspirin inhibits rotavirus replication and alters rat gut microbial composition.

BACKGROUND: Aspirin is widely used to treat various clinical symptoms. Evidence suggests that aspirin has antiviral properties, but little is known about its specific effect against rotavirus. METHODS: MA104, Caco-2, and CV-1 cells were infected with rotavirus, and aspirin was added after 12 h. Viral mRNA and titer levels were measured by qRT-PCR and immunofluorescence assays. For in vivo validation, forty specific-pathogen-free SD rats were randomly divided into oral aspirin (ASP) groups and control (NC) groups. 16 S rRNA gene sequencing was performed to identify gut microbiota. After 6 months of continuous ASP/NC administration, the rats were infected with rotavirus. Fecal samples were collected over a 30-day time course, and viral levels were quantified. Proinflammatory cytokines/chemokine levels were measured by ELISA. RESULTS: Aspirin inhibited rotavirus infection in cell lines and in rats. The effects of aspirin on viral replication were associated with the alteration of gut microbiota composition by aspirin, including increased abundance of Firmicutes and decreased abundance of Bacteroidetes after aspirin treatment. Mechanistically, aspirin reduced IL-2 and IL-10 levels, and increased IRF-1 and COX-2 levels. Aspirin blocked rotavirus replication in vitro and in vivo, which might be related to effects on IRF-1, COX-2, chemokines, and gut microbial composition. CONCLUSION: These results indicate that long-term oral aspirin administration reduces rotavirus infection. Intestinal virus infection may be suppressed in elderly patients who take aspirin for a long time. The change of their Gut microbiota may lead to functional disorder of the intestinal tract, which may provide some reference for clinical adjuvant probiotics treatment.

Successful treatment with doxycycline monotherapy for human infection with Babesia venatorum (Babesiidae, Sporozoa) in China: a case report and proposal for a clinical regimen.

BACKGROUND: Human babesiosis is a worldwide disease caused by intraerythrocytic protozoa of the genus Babesia. It is transmitted by bites from ixodid ticks, and mechanically transmitted by blood transfusion. It is primarily treated with quinine and/or atovaquone, which are not readily available in China. In this study, we developed a novel treatment regimen involving doxycycline monotherapy in a patient with severe Babesia venatorum infection as an alternative therapeutic medication. The aim of our study is to provide a guidance for clinical practice treatment of human babesiosis. CASE PRESENTATION: A 73-year-old man who had undergone splenectomy and blood transfusion 8 years prior, presented with an unexplained fever, headache, and thrombocytopenia, and was admitted to the Fifth Medical Center of the PLA General Hospital. He was diagnosed with B. venatorum infection by morphological review of thin peripheral blood smears, which was confirmed by multi-gene polymerase chain reaction (PCR), and sequencing of the entire 18s rRNA and partial ß-tubulin encoding genes, as well as isolation by animal inoculation. The doxycycline monotherapy regimen (peros, 0.1 g bisindie) was administered following pharmacological guidance and an effective outcome was observed. The patient recovered rapidly following the doxycycline monotherapy. The protozoan load in peripheral blood samples decreased by 88% in hematocrit counts after 8 days, and negative PCR results were obtained after 90 days of follow-up at the hospital. The treatment lasted for 3 months without any side effects or sequelae. The nine-month follow-up survey of the patient did not reveal any signs of recrudescence or anti-babesial tolerance. CONCLUSIONS: We have reported a clinical case of successful doxycycline monotherapy for human babesiosis caused by B. venatorum, which provides an optional medical intervention for human babesiosis.

Preoperative computed tomography-guided transscapular sens-cure needle localization for pulmonary nodule located behind the scapula.

BACKGROUND: Video-assisted thoracoscopic surgery (VATS) is an approach that is commonly used to resect pulmonary nodules (PNs). However, when these PNs are located behind the scapula, a transscapular access approach is generally required. In this study, the safety, efficacy, and feasibility of preoperative computed tomography (CT)-guided Sens-cure needle (SCN) localization was assessed for PNs located behind the scapula. METHODS: From January 2020 - June 2022, a total of 122 PN patients in our hospital underwent preoperative CT-guided SCN localization and subsequent VATS resection, of whom 12 (9.8%) exhibited PNs behind the scapula necessitating a transscapular approach for this localization procedure. RESULTS: This study included 12 patients, each of whom had one PN located behind the scapula. The CT-guided transscapular SCN localization approach was successful in all patients, and no complications near the operative site were observed. The median localization time was 12 min, and 2 (16.7%) and 1 (8.3%) patients respectively developed pneumothorax and pulmonary hemorrhage after the localization procedure was complete. Wedge resection procedures for these PNs achieved technical success in all cases. Four patients were diagnosed with invasive adenocarcinomas and subsequently accepted lobectomy and systematic lymph node dissection. The median VATS duration and the median blood loss was 80 min and 10 mL, respectively. In total, 3, 5, and 4 PNs were respectively diagnosed as benign, mini-invasive adenocarcinomas, and invasive adenocarcinomas. CONCLUSION: Preoperative CT-guided transscapular SCN localization represents a safe, straightforward, and effective means of localizing PNs present behind the scapula.

Differences and allometric relationships among assimilative branch traits of four shrubs in Central Asia.

Shrubs play a major role in maintaining ecosystem stability in the arid deserts of Central Asia. During the long-term adaptation to extreme arid environments, shrubs have developed special assimilative branches that replace leaves for photosynthesis. In this study, four dominant shrubs with assimilative branches, namely Haloxylon ammodendron, Haloxylon persicum, Calligonum mongolicum, and Ephedra przewalskii, were selected as the research objects, and the dry mass, total length, node number, and basal diameter of their assimilative branches and the average length of the first three nodes were carefully measured, and the allometric relationships among five traits of four species were systematically compared. The results indicated that: (1) Four desert shrubs have different assimilative branches traits. Compared with H. persicum and H. ammodendron, C. mongolicum and E. przewalskii have longer internodes and fewer nodes. The dry mass of H. ammodendron and the basal diameter of H. persicum were the smallest; (2) Significant allometric scaling relationships were found between dry mass, total length, basal diameter, and each trait of assimilative branches, all of which were significantly less than 1; (3) The scaling exponents of the allometric relationship between four traits and the dry mass of assimilative branches of H. persicum were greater or significantly greater than those of H. ammodendron. The scaling exponents of the relationships between the basal diameter, dry mass, and total length of E. przewalskii were higher than those of the other three shrubs. Therefore, although different species have adapted to drought and high temperatures by convergence, there was great variability in morphological characteristics of assimilative branches, as well as in the scaling exponents of relationships among traits. The results of this study will provide valuable insights into the ecological functions of assimilative branches and survival strategies of these shrubs to cope with aridity and drought in desert environments.

Clinical Analysis of Primary Tracheobronchial Tumors in Children and Evaluation of the Predicting Models for Mucoepidermoid Carcinoma.

OBJECTIVE: To determine the clinical characteristics and prognosis of primary tracheobronchial tumors (PTTs) in children, and to explore the most common tumor identification methods. METHODS: The medical records of children with PTTs who were hospitalized at the Children's Hospital of Chongqing Medical University from January 1995 to January 2020 were reviewed retrospectively. The clinical features, imaging, treatments, and outcomes of these patients were statistically analyzed. Machine learning techniques such as Gaussian naïve Bayes, support vector machine (SVM) and decision tree models were used to identify mucoepidermoid carcinoma (ME). RESULTS: A total of 16 children were hospitalized with PTTs during the study period. This included 5 (31.3%) children with ME, 3 (18.8%) children with inflammatory myofibroblastic tumors (IMT), 2 children (12.5%) with sarcomas, 2 (12.5%) children with papillomatosis and 1 child (6.3%) each with carcinoid carcinoma, adenoid cystic carcinoma (ACC), hemangioma, and schwannoma, respectively. ME was the most common tumor type and amongst the 3 ME recognition methods, the SVM model showed the best performance. The main clinical symptoms of PPTs were cough (81.3%), breathlessness (50%), wheezing (43.8%), progressive dyspnea (37.5%), hemoptysis (37.5%), and fever (25%). Of the 16 patients, 7 were treated with surgery, 8 underwent bronchoscopic tumor resection, and 1 child died. Of the 11 other children, 3 experienced recurrence, and the last 8 remained disease-free. No deaths were observed during the follow-up period. CONCLUSION: PTT are very rare in children and the highest percentage of cases is due to ME. The SVM model was highly accurate in identifying ME. Chest CT and bronchoscopy can effectively diagnose PTTs. Surgery and bronchoscopic intervention can both achieve good clinical results and the prognosis of the 11 children that were followed up was good.

A correlative study of iron metabolism based on q-Dixon MRI in benign prostatic hyperplasia and prostate cancer.

Clinical staging, Gleason score, and prostate-specific antigen (PSA) have been accepted as factors for evaluating the prognosis of prostate cancer (PCa). With the in-depth study of iron metabolism and the development of multiparametric magnetic resonance imaging technology, we used q-Dixon magnetic resonance imaging (MRI) to measure the iron content of the PCa patients' lesions, and used enzyme-linked immunosorbent assay (ELISA) to measure the iron metabolism indicators in the patients' serum samples, combined with the patients' postoperative clinical data for analysis. We found that the serum indexes were correlated with the T2 star values, International Society of Urological Pathology (ISUP) grade, and pathological classification in PCa patients (all P < 0.001) but not in benign prostatic hyperplasia (BPH) patients (all P > 0.05). The utilization of q-Dixon-based MRI and serum indexes allows the noninvasive measurement of iron content in prostate lesions and the assessment of differential iron metabolism between PCa and BPH, which may be helpful for evaluating the prognosis of PCa.

Assessment of peripheral neuropathy in type 2 diabetes by diffusion tensor imaging.

BACKGROUND: To evaluate the diagnostic accuracy of diffusion tensor imaging (DTI) in diabetic peripheral neuropathy (DPN) for patients with type 2 diabetes and detect the correlations with electrophysiology. METHODS: A total of 27 patients with type 2 diabetes with DPN, 24 patients with type 2 diabetes without peripheral neuropathy (NDPN), as well as 32 healthy controls (HC) were enrolled in this study. Clinical examinations and neurophysiologic tests were used to determine the presence of DPN. Fractional anisotropy (FA) and apparent diffusion coefficient (ADC) of peripheral nerves, including the tibial nerve (TN) and common peroneal nerve (CPN), were calculated. Receiver operating characteristic (ROC) analysis was performed for FA and ADC values. Pearson's correlation coefficient was used to assess the correlation between DTI and electrophysiology parameters in the patient group. RESULTS: The tibial and common peroneal nerve FAs were lowest (P=0.003, 0.001, respectively) and ADC was highest (P=0.004, 0.005, respectively) in the DPN group. The FA value of the axonal injury group was lower than that in the demyelination group (P=0.035, 0.01, respectively), while the ADC value was higher (P=0.02, 0.01, respectively). In the DPN group, FA value was positively correlated with motor conduction velocity (MCV) (tibial nerve: r=0.420, P=0.007; common peroneal nerve: r=0.581, P<0.001) and motor amplitude (MA) (tibial nerve: r=0.623, P<0.001; common peroneal nerve: r=0.513; P=0.001), while ADC values was negatively correlated with MCV (tibial nerve: r=-0.320, P=0.044; common peroneal nerve: r=-0.569; P<0.001), and MA (tibial nerve: r=-0.491, P=0.001; common peroneal nerve: r=-0.524; P=0.001). CONCLUSIONS: With a lower FA value and higher ADC value, DTI accurately discriminated DPN. The DTI multi-parameter quantitative analysis of peripheral nerves differentiated DPN axonal injury from the demyelinating lesion, and hence, could be applied in the diagnosis of DPN.

Analysis of the intestinal microbial community altered during rotavirus infection in suckling mice.

BACKGROUND: Rotavirus (RV) is a principal cause of diarrhea. However, there is a limited understanding regarding alteration of the gut microbial community structure and abundance during RV infection. This study was to characterize any potential associations between RV infection and the intestinal microbiota. METHODS: Suckling mice were divided into normal group (NC) and infected group (RV) randomly. All of the suckling mice were euthanized four days post-RV infection. The virus titer was counted as fluorescent focus assay, and viral load was quantified by QPCR. Five sucking mice were randomly selected from each RV group and NC group for sample collection and pathological analysis. Mixed intestinal contents of the colon and rectum were collected from all of the suckling mice. To investigate the detailed relationship between RV infection and intestinal microbiota, the composition and distribution of intestinal microbiota from suckling mice were first analyzed using 16S rRNA sequencing technology. RESULTS: The results of the pathological characteristics showed that vacuolar degeneration, vasodilation, hyperemia, and destruction of the intestinal epithelium were apparent in the RV group. Representative genera from Lactobacillus and Fusobacterium were enriched in the NC group, while the Enterococcus and Escherichia/Shigella genera were enriched in the RV group. Helicobacter, Alloprevotrlla, Brevundimonas, Paenibacillus, and Parabacteroides were completely undetectable in the RV group. The predicted intestinal flora metabolic function results showed that "carbohydrate metabolism" and "lipid metabolism" pathways were significantly enriched within the NC group. A significant difference has been observed in the gut microbiota composition between the two groups. CONCLUSIONS: Our results demonstrated a significant difference in the gut microbiota composition in RV-infected suckling mice as compared to the RV un-infected suckling mice group. This work may provide meaningful information regarding the bacterial genera changed during RV infection. Moreover, the changes in these bacteria may be related with the replication and pathogenesis of RV infection.

Iliac ecchymosis, a valuable sign for hollow viscus injuries in blunt pelvic trauma patients.

PURPOSE: Pelvic fractures are characterized by high energy injuries and often accompanied with abdominal and pelvic organ injury. CT has been applied for several decades to evaluate blunt pelvic trauma patients. However, it has a certain rate of inaccurate diagnosis of abdominal hollow viscus injury (HVI), especially in the early stage after injury. The delayed diagnosis of HVI could result in a high morbidity and mortality. The bowel injury prediction score (BIPS) applied 3 clinical variables to determine whether an early surgical intervention for blunt HVI was necessary. We recently found another clinical variable (iliac ecchymosis, IE) which appeared at the early stage of injury, could be predicted for HVI. The main objective of this study was to explore the novel combination of IE and BIPS to enhance the early diagnosis rate of HVI, and thus reduce complications and mortalities. METHODS: We conducted a retrospective analysis from January 2008 to December 2018 and recorded blunt pelvic trauma patients in our hospital. The inclusion criteria were patients who were verified with pelvic fractures using abdomen and pelvis CT scan in the emergency department before any surgical intervention. The exclusion criteria were abdominal CT insufficiency before operation, abdominal surgery before CT scan, and CT mesenteric injury grade being 5. The MBIPS was defined as BIPS plus IE, which was calculated according to 4 variables: white blood cell counts of 17.0 or greater, abdominal tenderness, CT scan grade for mesenteric injury of 4 or higher, and the location of IE. Each clinical variable counted 1 score, totally 4 scores. The location and severity of IE was also noted. RESULTS: In total, 635 cases were hospitalized and 62 patients were enrolled in this study. Of these included patients, 77.4% (40 males and 8 females) were operated by exploratory laparotomy and 22.6% (8 males and 6 females) were treated conservatively. In the 48 patients underwent surgical intervention, 46 were confirmed with HVI (45 with IE and 1 without IE). In 46 patients confirmed without HVI, only 3 patients had IE and the rest had no IE. The sensitivity and specificity of IE in predicting HVI was calculated as 97.8% (45/46) and 81.3% (13/16), respectively. The median MBIPS score for surgery group was 2, while 0 for the conservative treatment group. The incidence of HVI in patients with MBIPS score ≥ 2 was significantly higher than that in patients with MBIPS score less than ≤ 2 (OR = 17.3, p < 0.001). CONCLUSION: IE can be recognized as an indirect sign of HVI because of the high sensitivity and specificity, which is a valuable sign for HVI in blunt pelvic trauma patients. MBIPS can be used to predict HVI in blunt pelvic trauma patients. When the MBIPS score is ≥ 2, HVI is strongly suggested.

Gait analysis in the elderly patients with lumbar spinal stenosis.

PURPOSE: This study aims to analyze the gait characteristics of the elderly patients with lumbar spinal stenosis by an intelligent device for energy expenditure and activity (IDEEA) to assist clinical work. METHODS: A total of 98 subjects were included in this study from January 2017 to December 2018. A total of 49 elderly outpatients with symptomatic lumbar spinal stenosis in unilateral lower extremity were included as the experimental group, and another 49 healthy subjects matched with gender, age, and body mass index (BMI) were analyzed as the control group. The gait data of the subjects (including single support, double support, SLS/DLS, swing duration, step duration, cycle duration, pulling accel, swing power, ground impact, foot fall, foot off, push off, speed, cadence, step length, and stride length) were collected to compare between the experience group and control group, the affected leg and the healthy leg in experimental group. RESULTS: The results of this study presented that small intermittent claudication occurred in all patients. The time of single support was significantly increased (p < 0.05). Double support, step duration, and pulling accel were increased (p < 0.05), and the Push off, speed, step length, and Stride length were decreased (p < 0.05) in the experimental group compared with the control group. CONCLUSION: Small intermittent claudication was the basic gait composition of the elderly patients with lumbar spinal stenosis that can reflect the abnormal gait characteristics by IDEEA.

Characterization of a G9 group A rotavirus reassortant strain detected in Jinzhou, China, in 2018-2019.

A novel rotavirus A (RVA) strain (JZ) was detected in RVA-positive stool specimens from three pediatric patients in Jinzhou, Liaoning Province, in 2018-2019. The electrophoresis pattern of the JZ strain showed a long electropherotype. The genomic constellation G9-P[8]-I1-R1-C1-M1-A1-N1-T1-E2-H1 was detected, suggesting that a new inter-genogroup reassortment had occurred. Whole-genome sequencing showed that the JZ isolates were identical to each other. Phylogenetic analysis revealed that VP7 and VP4 clustered in lineages G9-VI and P[8]-3, respectively. JZ strain-specific amino acid substitutions were detected in VP7, VP4 and NSP4. This study provides information on the epidemiology and characteristics of G9 strains circulating in China.

The effects of autophagy on the replication of Nelson Bay orthoreovirus.

BACKGROUND: Nelson Bay orthoreovirus (NBV) was first isolated over 40 years ago from a fruit bat in Australia. Normally, NBV does not cause human diseases, but recently several NBV strains have been associated with human respiratory tract infections, thus attracting clinical attention. Autophagy, an evolutionarily conserved process in eukaryotic cells, degrades intracellular substrates, participates in multiple physiological processes, and maintains cellular homeostasis. In addition, autophagy is intimately involved in viral infection. METHODS: A new strain of NBV, isolated from a patient with a respiratory tract infection who returned to Japan from Bali, Indonesia, in 2007, was used in this study. NBV was rescued using a reverse genetics system involving cotransfection of BHK cells with 11 plasmids (pT7-L1 MB, pT7-L2 MB, pT7-L3 MB, pT7-M1 MB, pT7-M2 MB, pT7-M3 MB, pT7-S1 MB, pT7-S2 MB, pT7-S3 MB, pT7-S4 MB, and pcDNA3.1-T7), yielding NBV-MB. Recovered viruses were confirmed by immunofluorescence. The effect of NBV-MB on autophagy was evaluated by measuring the LC3-I/II proteins by immunoblot analysis after infection of BHK cells. Furthermore, after treatment with rapamycin (RAPA), 3-methyladenine (3-MA), chloroquine (CQ), or plasmid (GFP-LC3) transfection, the changes in expression of the LC3 gene and the amount of LC3-I/II protein were examined. In addition, variations in viral titer were assayed after treatment of BHK cells with drugs or after transfection with plasmids pCAGM3 and pCAGS3, which encode virus nonstructural proteins µNS and σNS, respectively. RESULTS: NBV-MB infection induced autophagy in host cells; however, the level of induction was dependent on viral replication. Induction of autophagy increased viral replication. By contrast, inhibiting autophagy suppressed NBV replication, albeit not significantly. The NBV-MB nonstructural protein µNS was involved in the induction of autophagy with viral infection. CONCLUSIONS: NBV-MB infection triggered autophagy. Also, the NBV nonstructural protein µNS may contribute to augmentation of autophagy upon viral infection.

Coronary artery anatomy in peri-crux cordis area on computed coronary tomography angiography.

BACKGROUND: The peri-crux area is an anatomical structure of the heart. Unfortunately, important information on this area mainly derives from autopsy heart with a small, under-representative sample size, resulting in limited clinical applications. Furthermore, little has been done to standardize the definition of the peri-crux area on coronary computed tomography angiography (CCTA) images or to investigate coronary artery anatomy wherein potential values are attracting experienced inventional cardiologists in terms of the revascularization strategies. The current study aimed to identify the peri-crux cordis area and to observe coronary artery anatomical distributions in this area on CCTA. METHODS: A total of 1,006 consecutive patients undergoing CCTA exams were enrolled. We delineated the peri-crux cordis area based on the posterior interatrial sulcus, posterior interventricular sulcus (PIS), left and right posterior atrioventricular groove on the diaphragmatic surface of the heart. Then we observed the coronary artery distributions in the peri-crux cordis area in different sexes. RESULTS: We have defined the peri-crux cordis area according to the anatomical landmarks on the diaphragmatic surface of the heart on CCTA images. We have observed 8 coronary artery distributions in the peri-crux cordis area. Right dominance has 4 types (types 1-4); left, 1 type (type 0) and balanced, 3 types (types 5-7). Out of the 1,006 cases, the type 1 is commonest with 834 cases (82.9%). There are no statistically significant differences in terms of coronary dominances and coronary artery distributions in the peri-crux cordis area between sexes (P>0.05). CONCLUSIONS: We have defined the peri-crux cordis area utilizing the anatomical landmarks of the heart on CCTA images, where 8 types of coronary artery distributions have been identified. The current study may provide interventional cardiologists with useful information on recognition of coronary artery dominance, use of collateral channels for revascularization of chronic total occluded lesions, and evaluation of prognosis in patients with coronary artery disease (CAD).

Host protein CD63 promotes viral RNA replication by interacting with human astrovirus non-structural protein nsP1a/4.

Human astrovirus non-structural protein nsP1a/4, located at the C-terminal end of nsP1a, is thought to be involved in regulating RNA replication. Here, we show that host protein CD63 interacts with the nsP1a protein. Further research showed that the large loop (LEL) domain of CD63 also interacts with nsP1a/4. Confocal microscopy showed that nsP1a/4 protein and CD63 co-localized in the cytoplasm of co-transfected cells. Co-localization of nsP1a/4 and CD63 was also observed in HAstV-1-infected cells. Overexpression of CD63 promoted replication of HAstV-1, whereas knockdown of CD63 reduced production of HAstV-1 viral progeny. These results suggest that CD63 plays a critical role in HAstV-1 replication, and provide an avenue to further understanding the interactions between host and virus proteins during replication and pathogenesis of HAstV.

Weighted Gene Correlation Network Analysis (WGCNA) Detected Loss of MAGI2 Promotes Chronic Kidney Disease (CKD) by Podocyte Damage.

BACKGROUND/AIMS: Podocyte damage is associated with proteinuria, glomerulosclerosis and decline of renal function. This study aimed to screen critical genes associated with podocyte injury in chronic kidney disease (CKD) by weighted gene correlation network analysis (WGCNA), and explore related functions. METHODS: GSE66107, GSE93798, GSE30528, GSE32591 gene expression data including podocyte injury models or glomeruli in CKD patients were downloaded from the GEO database. R was used for data analysis. Differentially expressed genes (DEGs) (FDR< 0.05 or |Fold Change|≥1.5) in GSE993395 were assessed by WGCNA. According to Gene Ontology (GO) and known podocyte standard genes (PSGs), podocyte injury-associated modules were defined, with hub genes selected based on average intramodular connectivity. The Cytoscape software was used for network visualization. Nephroseq was used to assess the clinical significance of hub genes. Small interfering RNA (siRNA) was used to evaluate the roles of hub genes in podocyte injury Results: Totally 7957 DEGs were screened, with 15 (co.DEGs) altered in all 4 datasets; 4031 DEGs were used for WGCNA, encompassing 12 modules. Green modules (most PSGs and co.DEGs) were significantly enriched in glomerular development, and considered podocyte injury-associated modules. Furthermore, MAGI2 (a hub gene) was also a co.DEG and PSG. Glomerular MAGI2 levels were reduced in various kidney diseases, and positively and negatively associated with glomerular filtration rate and urinary protein levels in CKD patients. Moreover, MAIG2 knockdown reduced NPHS2, CD2AP and SYNPO levels, and induced podocyte rearrangement and apoptosis. CONCLUSION: MAGI2 identified by WGCNA regulates cytoskeletal rearrangement in podocytes, with its loss predisposing to proteinuria and CKD.

In vitro effect of Porphyromonas gingivalis combined with influenza A virus on respiratory epithelial cells.

OBJECTIVE: Respiratory epithelial cells are the first natural barrier against bacteria and viruses; hence, the interactions among epithelial cells, bacteria, and viruses are associated with disease occurrence and development. The effect of co-infection by P. gingivalis and influenza A virus (IAV) on respiratory epithelial cells remains unknown. The aim of this study was to analyze in vitro cell viability and apoptosis rates in respiratory epithelial A549 cells infected with P. gingivalis or IAV alone, or a combination of both pathogens. DESIGN: A549 cells were first divided into a control group, a P. gingivalis group, an IAV group, and a P. gingivalis + IAV group, to examine cell viability and apoptosis rates, the levels of microtubule associated protein 1 light chain 3 B (LC3-II), microtubule associated protein 1 light chain 3A (LC3-I), and sequestosome 1 (P62), and the formation of autophagosomes. The autophagy inhibitor, 3-methyladenine (3MA), was used to assess autophagy and apoptosis in A549 cells infected with P. gingivalis or IAV. RESULTS: An MTT assay revealed that cell viability was significantly lower in the IAV group than in the P. gingivalis + IAV group (P < 0.05). Flow cytometry indicated that the apoptosis rate was significantly higher in the IAV group than in the P. gingivalis + IAV group (P < 0.05). The fluorescence levels of GFP-LC3 increased significantly, the LC3-II/LC3-I ratio was significantly higher, and the P62 protein levels were statistically lower in the P. gingivalis + IAV group compared with the IAV group (all P < 0.05). Western blotting revealed that the LC3- II/LC3-I ratio was significantly lower, and caspase-3 levels were significantly higher in the 3MA + P. gingivalis + IAV group compared to the P. gingivalis + IAV group (all P < 0.05). CONCLUSION: In vitro studies showed that infection by P. gingivalis combined with IAV temporarily inhibited apoptosis in respiratory epithelial cells, and this may be related to the initiation of autophagy.

Effect of Period 2 on the proliferation, apoptosis and migration of osteosarcoma cells, and the corresponding mechanisms.

Period 2 (per2) is a core circadian clock gene. Dysregulation of the per2 gene has been identified in a number of types of human cancer and may be associated with a poor prognosis. To confirm the influence of per2 gene on MNNG/HOS human osteosarcoma cells, small interfering (si)RNA against per2 or plasmids containing per2 were transfected into MNNG/HOS cells, and the proliferation, apoptosis and migration were observed. The present study demonstrated that per2 knockdown significantly enhanced MNNG/HOS cell proliferation and migration and protected MNNG/HOS cells from apoptosis. Per2 overexpression inhibited MNNG/HOS cell proliferation and migration and promoted apoptosis. Furthermore, the protein expression of phosphorylated (p)-protein kinase B (Akt) and Bcl-2 were inhibited in per2-overexpressing cells, while the expression of p27, p21 and cleaved caspase-3 was promoted. In contrast, the expression of p-Akt and Bcl-2 was promoted in per2-knockdown cells, and p27, p21 and cleaved caspase-3 were decreased. This initial study may provide an alternative therapeutic strategy for the treatment of osteosarcoma.