Integrated analysis of single-cell and bulk RNA sequencing data identifies the characteristics of ferroptosis in lumbar disc herniation.

Lumbar disc herniation (LDH) is the most common condition associated with low back pain, and it adversely impacts individuals' health. Ferroptosis has recently emerged as a novel factor in the pathogenesis of LDH; however, the specific impacts of ferroptosis on LDH have not been fully elucidated. Ferroptosis-related differentially expressed genes (FRDEGs) were identified from the transcriptomic datasets of LDH. Gene set enrichment analysis (GSEA) was conducted to identify biological mechanism and related pathways. LASSO and SVM-RFE algorithms were applied to detect signature genes. Function of the signature gene was confirmed by RT-qPCR. The CIBERSORT algorithm was used to compare immune infiltration between LDH and normal samples. Correlation analysis between MYB and immune cells was analyzed using the Pearson method. Additionally, we used scRNA-seq to dissect cell clusters and cellular interactions. AUCell scoring was used to analyze the ferroptosis scores of different cell types. We found that MYB, a highly expressed ferroptosis-related gene, was associated with LDH By leveraging bioinformatics analysis. In immune infiltration analysis, the abundance of monocytes and macrophages varied significantly between the LDH group and disc spondylolisthesis (DS) group. MYB was correlated with most immune cells. GSEA revealed MYB was significantly enriched in immune-related pathways. Furthermore, scRNA-seq analysis revealed the presence of eight distinct cell types. AUCell analysis showed that macrophages had a high ferroptosis score. Cell trajectory analysis revealed that chondrocyte 1 was at the beginning of the trajectory, while calcification inhibiting chondrocytes and fibrochondrocytes accumulated along the middle and tail end of the trajectory, respectively. Cell-cell communication analysis identified chondrocyte 1 had an extensive communication network with other clusters and interacted with nucleus pulposus through collagen signaling pathway. Our analysis demonstrated that MYB may be a potential therapeutic target for LDH. This study provides a resource for the orthopedics community that will facilitate additional discoveries directedly toward understanding the pathogenesis process of LDH.

Estradiol is a key candidate for treating Ankylosing Spondylolisthesis with Traditional Chinese Medicine.

Some Traditional Chinese Medicine (TCM) has shown anti-inflammatory and immunosuppressive effects on Ankylosing Spondylitis (AS) treatment. Wan Bikang (WBK) and Wan Biqing (WBQ) are two traditional empirical formulas for AS. However, the mechanism of their effects on AS is largely unknown. This study deciphered the underlying common molecular mechanisms of these TCM treatments for AS. The ultra-high-performance liquid chromatography-triple/time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS) assays were employed to detect herbal ingredients. Target proteins of herbal ingredients were identified by ChEMBL Database. To infer the relationships between ingredients and AS-related proteins, network pharmacology was employed. Protein-protein interaction (PPI) network and core target analyses were carried out with tools Cytoscape and STRING. To find out the molecular basis and target of AS, molecular docking and an in vitro experiment were also conducted. It is found that estradiol may participate in the treatment of AS via the inhibition of inflammatory factors, and Estrogen Receptor 1 (ESR1) appears to be a key target. This research offers insight into the therapeutic mechanism of TCM formulas for AS and furthers our understanding of TCM pharmacology.

scRNA-seq and proteomics reveal the distinction of M2-like macrophages between primary and recurrent malignant glioma and its critical role in the recurrence.

AIMS: Tumor-associated macrophages (TAMs) in the immune microenvironment play an important role in the increased drug resistance and recurrence of malignant glioma, but the mechanism remains incompletely inventoried. The focus of this study was to investigate the distinctions of M2-like TAMs in the immune microenvironment between primary and recurrent malignant glioma and its influence in the recurrence. METHODS: We employed single-cell RNA sequencing to construct a single-cell atlas for a total of 23,010 individual cells from 6 patients with primary or recurrent malignant glioma and identified 5 cell types, including TAMs and malignant cells. Immunohistochemical techniques and proteomics analysis were performed to investigate the role of intercellular interaction between malignant cells and TAMs in the recurrence of malignant glioma. RESULTS: Six subgroups of TAMs were annotated and M2-like TAMs were found to increase in recurrent malignant glioma significantly. A pseudotime trajectory and a dynamic gene expression profiling during the recurrence of malignant glioma were reconstructed. Up-regulation of several cancer pathways and intercellular interaction-related genes are associated with the recurrence of malignant glioma. Moreover, the M2-like TAMs can activate the PI3K/Akt/HIF-1α/CA9 pathway in the malignant glioma cells via SPP1-CD44-mediated intercellular interaction. Interestingly, high expression of CA9 can trigger the immunosuppressive response in the malignant glioma, thus promoting the degree of malignancy and drug resistance. CONCLUSION: Our study uncovers the distinction of M2-like TAMs between primary and recurrent glioma, which offers unparalleled insights into the immune microenvironment of primary and recurrent malignant glioma.

Allergic disease and sensitization disparity in urban and rural China: A EuroPrevall-INCO study.

BACKGROUND: Studies in comparison with allergic diseases and sensitization between rural and urban environments in westernized countries might be biased and not adequately reflect countries undergoing rapid transition. METHODS: A total of 5542 schoolchildren from urban area and 5139 from rural area were recruited for the EuroPrevall-INCO survey. A subsequent case-control sample with 196 children from urban area and 202 from rural area was recruited for a detailed face-to-face questionnaire and assessment of sensitization. Skin prick tests and serum-specific IgE measurements were used to assess sensitizations against food and aeroallergens. Logistic regression analysis was used to determine associations between risk/protective factors, food adverse reactions (FAR), allergic diseases, and sensitizations. RESULTS: Prevalence of self-reported allergic diseases, including asthma (6.6% vs.2.5%), rhinitis (23.2% vs.5.3%), and eczema (34.1% vs.25.9%), was higher in urban than in rural children. Urban children had a significantly higher prevalence of FAR and related allergic diseases, and lower food/inhalation allergen sensitization rate, than those of rural children. In urban children, frequent changing places of residency (odds ratio 2.85, 95% confidence interval: 1.45-5.81) and antibiotic usage (3.54, 1.77-7.32) in early life were risk factors for sensitization, while sensitization and family history of allergy were risk factors for allergic diseases. In rural children, exposure to rural environments in early life was protective against both allergen sensitizations (0.46, 0.21-0.96) and allergic diseases (0.03, 0.002-0.19). CONCLUSION: We observed a disparity in rates of allergic diseases and allergen sensitization between rural and urban children. In addition to family history, the development of allergic diseases and allergen sensitization were associated with specific urban/rural environmental exposures in early life.

Rural environment reduces allergic inflammation by modulating the gut microbiota.

Rural environments and microbiota are linked to a reduction in the prevalence of allergies. However, the mechanism underlying the reduced allergies modulated by rural residency is unclear. Here, we assessed gut bacterial composition and metagenomics in urban and rural children in the EuroPrevall-INCO cohort. Airborne dusts, including mattress and rural henhouse dusts, were profiled for bacterial and fungal composition by amplicon sequencing. Mice were repeatedly exposed to intranasal dust extracts and evaluated for their effects on ovalbumin (OVA)-induced allergic airway inflammation, and gut microbiota restoration was validated by fecal microbiota transplant (FMT) from dust-exposed donor mice. We found that rural children had fewer allergies and unique gut microbiota with fewer Bacteroides and more Prevotella. Indoor dusts in rural environments harbored higher endotoxin level and diversity of bacteria and fungi, whereas indoor urban dusts were enriched with Aspergillus and contained elevated pathogenic bacteria. Intranasal administration of rural dusts before OVA sensitization reduced respiratory eosinophils and blood IgE level in mice and also led to a recovery of gut bacterial diversity and Ruminiclostridium in the mouse model. FMT restored the protective effect by reducing OVA-induced lung eosinophils in recipient mice. Together, these results support a cause-effect relationship between exposure to dust microbiota and allergy susceptibility in children and mice. Specifically, rural environmental exposure modulated the gut microbiota, which was essential in reducing allergy in children from Southern China. Our findings support the notion that the modulation of gut microbiota by exposure to rural indoor dust may improve allergy prevention.

Identification of JUN as determinant of osteoarthritis and its inhibition by the Chinese herbal formulae Zhuanggu Huoxue Tang.

Synovitis is an essential feature of Osteoarthritis (OA). Increasing evidence demonstrates that synovitis plays a critical role in OA's symptoms and structural progression. However, there is no effective drug for preventing and treating synovitis. Some Chinese herbal formulae have been found to treat clinical OA effectively, however, their mode of action is still unclear. This study investigated the Chinese herbal formulae Zhuanggu Huoxue Tang (ZHT) underlying mechanisms for treating osteoarthritis. Transcriptome data and a network pharmacology analysis were used to investigate the biochemical pathways affected by ZHT during OA treatment with in vitro verification. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were undertaken. The interaction network of the ZHT active constituent targets was determined using Cytoscape 3.7.1 software. Molecular docking of key pathogenic proteins and components of ZHT was performed in silico to confirm the compounds' pharmaceutical activities. The results establish that JUN is a target pathway in the pathogenesis of osteoarthritis. Miltirone, one of the active ingredients of ZHT, demonstrated a suitable binding activity with JUN. Miltirone alleviates the catabolic gene expression induced by IL-1ß and IL-6 in synovial fibroblasts (FLS), validating the use of Miltirone as a therapeutic drug for osteoarthritis.

Characterization of a Novel Model of Lumbar Ligamentum Flavum Hypertrophy in Bipedal Standing Mice.

OBJECTIVE: To explore the main causes of hypertrophied ligamentum flavum (HLF) and the possibility of using bipedal standing mouse model to simulate the pathological changes in human HLF. METHODS: Thirty-two 8-week-old C57BL/6 male mice were randomly assigned to the experimental group (n = 16) and control group (n = 16). In the experimental group, mice were induced to adopt a bipedal standing posture by their hydrophobia. The experimental mice were maintained bipedal standing for 8 h a day with an interval of 2 h to consume food and water. The control mice were placed in a similar environment without bipedal standing. Eight 18-month-old C57BL/6 male mice were compared to evaluate the LF degeneration due to aging factor. Three-dimensional (3D) reconstruction and finite element models were carried out to analyze the stress and strain distribution of the mouse LF in sprawling and bipedal standing postures. Hematoxylin and Eosin (HE), Verhoeff-Van Gieson (VVG), and immunohistochemistry (IHC) staining were used to evaluate the LF degeneration of mice and humans. RT-qPCR and immunofluorescence analysis were used to evaluate the expressions of fibrosis-related factors and inflammatory cytokines of COL1A1, COL3A1, α-SMA, MMP2, IL-1ß, and COX-2. RESULTS: The von Mises stress (8.85 × 10-2 MPa) and maximum principal strain (6.64 × 10-1 ) in LF were increased 4944 and 7703 times, respectively, in bipedal standing mice. HE staining showed that the mouse LF area was greater in the bipedal standing 10-week-old group ([10.01 ± 2.93] × 104 µm2 ) than that in the control group ([3.76 ± 1.87] × 104 µm2 ) and 18-month-old aged group ([6.09 ± 2.70] × 104 µm2 ). VVG staining showed that the HLF of mice (3.23 ± 0.58) and humans (2.23 ± 0.31) had a similar loss of elastic fibers and an increase in collagen fibers. The cell density was higher during the process of HLF in mice (39.63 ± 4.81) and humans (23.25 ± 2.05). IHC staining showed that the number of α-SMA positive cells were significantly increased in HLF of mice (1.63 ± 0.74) and humans (3.50 ± 1.85). The expressions of inflammatory cytokines and fibrosis-related factors of COL1A1, COL3A1, α-SMA, MMP2, IL-1ß, and COX-2 were consistently higher in bipedal standing group than the control group. CONCLUSION: Our study suggests that 3D finite element models can help analyze the abnormal stress and strain distributions of LF in modeling mice. Mechanical stress is the main cause of hypertrophied ligamentum flavum compared to aging. The bipedal standing mice model can reflect the pathological characteristics of human HLF. The bipedal standing mice model can provide a standardized condition to elucidate the molecular mechanisms of mechanical stress-induced HLF in vivo.

miR-10396b-3p inhibits mechanical stress-induced ligamentum flavum hypertrophy by targeting IL-11.

Ligamentum flavum hypertrophy (LFH) leads to lumbar spinal stenosis (LSS) caused by LF tissue inflammation and fibrosis. Emerging evidence has indicated that dysregulated microRNAs (miRNAs) have an important role in inflammation and fibrosis. Mechanical stress (MS) has been explored as an initiating step in LFH pathology progression; the inflammation-related miRNAs induced after mechanical stress have been implicated in fibrosis pathology. However, the pathophysiological mechanism of MS-miRNAs-LFH remains to be elucidated. Using miRNAs sequencing analysis and subsequent confirmation with qRT-PCR assays, we identified the decreased expression of miR-10396b-3p and increased expression of IL-11 (interleukin-11) as responses to the development of LSS in hypertrophied LF tissues. We also found that IL-11 is positively correlated with fibrosis indicators of collagen I and collagen III. The up-regulation of miR-10396b-3p significantly decreased the level of IL-11 expression, whereas miR-10396b-3p down-regulation increased IL-11 expression in vitro. Luciferase reporter assay indicates that IL-11 is a direct target of miR-10396b-3p. Furthermore, cyclic mechanical stress inhibits miR-10396b-3p and induces IL-11, collagen I, and collagen III in vitro. Our results showed that overexpression of miR-10396b-3p suppresses MS-induced LFH by inhibiting collagen I and III via the inhibition of IL-11. These data suggest that the MS-miR-10396b-3p-IL-11 axis plays a key role in the pathological progression of LFH.

Impurity-driven transitions in frustrated quantum Ising rings.

We study the quantum phase transition driven by a point impurity in a chain seamed with ring frustration. With strong coupling and light impurity, the system is in a topological extended-kink (TEK) phase, which exhibits gapless excitations in the bulk. With strong coupling and heavy impurity, the system is in a gapped kink bound state (KBS) phase. Two-point bulk and impurity correlations are defined to characterize the two phases. In the TEK phase, both the bulk and impurity correlations are long range and factorizable so that scaling functions can be parsed. The scaling functions relies on the distance scaled by the system's size. An impurity correlation length can be extracted from the impurity correlation. In the transition from TEK to KBS, the scaling function of the bulk correlation undergoes an abrupt steplike change. Meanwhile, the impurity correlation length decreases from a divergent value to a finite one. The ground state of the TEK phase retains a relatively high value of entanglement entropy due to the absence of symmetry breaking. However, spontaneous symmetry breaking occurs in the KBS phase, which induces antiferromagnetic order in the bulk and entangled spin configuration near the impurity.

CRLF1 Is a Key Regulator in the Ligamentum Flavum Hypertrophy.

Hypertrophy of the ligamentum flavum (HLF) is one of the common causes of lumbar spinal stenosis (LSS). The key molecules and mechanisms responsible for HLF remain unclear. Here, we used an integrated transcriptome and proteomics analysis of human ligamentum flavum (LF), and subsequent immunohistochemistry and real-time PCR assays, to show upregulation of CRLF1 to be the dominant response to HLF. TGF-ß1 significantly increased mRNA expression of CRLF1 through SMAD3 pathway. CRLF1 enhanced LF fibrosis via ERK signaling pathway at the post-transcriptional level and was required for the pro-fibrotic effect of TGF-ß1. Knockdown of CRLF1 was shown here to reduce fibrosis caused by inflammatory cytokines and mechanical stress. Furthermore, we found that bipedal standing posture can cause HLF and upregulation of CRLF1 expression in mice LF. Overexpression of CRLF1 was indicated to cause HLF in vivo, whereas CRLF1 knockdown impeded the formation of HLF in bipedal standing mice. These results revealed a crucial role of CRLF1 in LF hypertrophy. We propose that inhibition of CRLF1 is a potential therapeutic strategy to treat HLF.

Deep Learning Algorithm Classifies Heartbeat Events Based on Electrocardiogram Signals.

Cardiovascular diseases (CVDs) have become the number 1 threat to human health. Their numerous complications mean that many countries remain unable to prevent the rapid growth of such diseases, although significant health resources have been invested toward their prevention and management. Electrocardiogram (ECG) is the most important non-invasive physiological signal for CVD screening and diagnosis. For exploring the heartbeat event classification model using single- or multiple-lead ECG signals, we proposed a novel deep learning algorithm and conducted a systemic comparison based on the different methods and databases. This new approach aims to improve accuracy and reduce training time by combining the convolutional neural network (CNN) with the bidirectional long short-term memory (BiLSTM). To our knowledge, this approach has not been investigated to date. In this study, Database I with single-lead ECG and Database II with 12-lead ECG were used to explore a practical and viable heartbeat event classification model. An evolutionary neural system approach (Method I) and a deep learning approach (Method II) that combines CNN with BiLSTM network were compared and evaluated in processing heartbeat event classification. Overall, Method I achieved slightly better performance than Method II. However, Method I took, on average, 28.3 h to train the model, whereas Method II needed only 1 h. Method II achieved an accuracy of 80, 82.6, and 85% compared with the China Physiological Signal Challenge 2018, PhysioNet Challenge 2017, and Massachusetts Institute of Technology-Beth Israel Hospital (MIT-BIH) Arrhythmia datasets, respectively. These results are impressive compared with the performance of state-of-the-art algorithms used for the same purpose.

A specific aggregation-induced emission-conjugated polymer enables visual monitoring of osteogenic differentiation.

Osteogenic differentiation is the basis of bone growth and repair related to many diseases, in which evaluating the degree and ability of osteogenic transformation is quite important and highly desirable. However, fixing or stopping the growth of cells is required for conventional methods to monitor osteogenic differentiation, which cannot realize the full investigation of the dynamic process. Herein, a new anion conjugated polymer featuring aggregation-induced emission (AIE) characteristics is developed with excellent solubility for in-situ monitoring the process of osteogenic differentiation. This novel polymer can bind with osteogenic differentiated cells, and the intracellular fluorescence increases gradually with the enhancement of osteogenic differentiation. Moreover, it possesses good biosafety with negligible effect on cell activity and osteogenic differentiation, which cannot be realized by the typical method of Alizarin Red S staining. Further study shows that the polymer crosses the cell membrane through endocytosis and enriches in lysosomes, whereas no obvious fluorescence is detected with other cells, including non-differentiated osteoblast cells, under the same conditions, demonstrating the high selectivity. This is the first fluorescent probe with excellent specificity to realize real-time observation of the process of osteogenic differentiation. Therefore, PTB-EDTA shows great promise in the study of osteogenic differentiation and related applications.

Relationship between the location of ligamentum flavum hypertrophy and its stress in finite element analysis.

OBJECTIVE: To quantitatively describe the stress of the ligamentum flavum (LF) using the finite element method and to compare the stress at different parts of the healthy LF. METHODS: Based on the high resolution computed tomography imaging data of a healthy 22-year-old man, three-dimensional nonlinear L4-5 lumbar finite element model (FEM) representing intact condition was developed. The LF, as the object of the present research, was incorporated into the spinal model in the form of solid three-dimensional structure. The model's validity is verified by comparing its biomechanical indices, such as range of motion and axial compression pressure displacement, with published results under specific loading conditions. To authenticate the accuracy of the solid LF, the lamina attachments, the central cross-section, and other anatomy indicators were compared with figures in the published literature. After the average and maximum von Mises stress on the surface of LF under various working conditions were measured using ANSYS and AutoCAD software, the surface stress difference in the LF between the ventral and dorsal sides as well as the lateral and lamina parts were determined. RESULTS: The FEM predicted a similar tendency for biomechanical indices as shown in previous studies. The lamina attachments, the central cross-section, and the height as well as the width of the LF in the healthy FEM were in accordance with published results. In the healthy model, the average and maximum von Mises stress in the shallow layer of the LF were, respectively, 1.40, 2.28, 1.76, 1.48, 1.38 and 1.79, 2.41, 1.46, 1.42, 1.71 times that in the deep layer under a compressive preload of 500 N incorporated with flexion, extension, and lateral and rotational moments (10 Nm). The most conspicuous difference in surface stress was observed with the flexion motion, with a nearly 241% difference in the maximum stress and a 228% difference in the average stress compared to those in other states. As far as the whole dorsal side of the LF was concerned, the maximum surface stress was almost all concentrated in the dorsal neighboring facet joint portion. In addition, the maximum and average stress were, respectively, 77%, 72%, 15%, 11%, 71% and 153%, 39%, 54%, 200%, 212% higher in the lateral part than in the lamina part. CONCLUSION: Based on the predisposition of LF hypertrophy in the human spine and the stress distribution of this study, the positive correlation between LF hypertrophy and its stress was confirmed.

Casticin Attenuates Osteoarthritis-Related Cartilage Degeneration by Inhibiting the ROS-Mediated NF-κB Signaling Pathway in vitro and in vivo.

Casticin, a flavonoid isolated from Vitex trifolia, has been shown to have anti-inflammatory and antitumor effects in previous studies. Osteoarthritis (OA) is a disease based on degenerative pathological changes. The disease process is often accompanied by inflammatory pathological changes. However, there is no safe and effective drug for prevention and treatment. In the present study, we aimed to clarify the role of casticin in the murine model of destabilization of the medial meniscus (DMM). Male BALB/c mice were randomly divided into three groups: Sham, DMM-induced OA treated with vehicle, and DMM-induced OA treated with casticin. Our results indicated that the casticin treatments markedly reduced the destruction of cartilage and OARSI grades compared with those of the vehicle-treated mice. The levels of matrix metalloproteinase-13 (MMP13) in cartilage were also significantly reduced in the casticin-treated mice. Casticin also significantly regulated oxidative stress and reduced inflammation in the cartilage of mice with OA. These results suggest that casticin prevents the development of posttraumatic OA in mice. Consequently, decreased reactive oxygen species levels and suppressed proinflammatory cytokine production were confirmed in casticin-treated IL-1ß-stimulated ADTC5 cells. After casticin treatment, the NF-κB signaling pathway was significantly inhibited in the cells. It can be concluded that casticin can alleviate arthritis-related cartilage degeneration by inhibiting ROS-mediated NF-κB signaling pathway in vitro and in vivo.

Rigorous proof for the nonlocal correlation function in the transverse Ising model with ring frustration.

An unusual correlation function was conjectured by Campostrini et al. [Phys. Rev. E 91, 042123 (2015)PLEEE81539-375510.1103/PhysRevE.91.042123] for the ground state of a transverse Ising chain with geometrical frustration. Later, we provided a rigorous proof for it and demonstrated its nonlocal nature based on an evaluation of a Toeplitz determinant in the thermodynamic limit [J. Stat. Mech. (2016) 11310210.1088/1742-5468/2016/11/113102]. In this paper, we further prove that all the low excited energy states forming the gapless kink phase share the same asymptotic correlation function with the ground state. As a consequence, the thermal correlation function almost remains constant at low temperatures if one assumes a canonical ensemble.

Using positive-ion electrospray ionization mass spectrometry and H/D exchange study phosphoryl group transfer reactions involved in amino acid ester isopropyl phosphoramidates of Brefeldin A.

As mini-chemical models, amino acid ester isopropyl phosphoramidates of Brefeldin A (compounds 2a-2d) were synthesized and investigated by electrospray ionization tandem mass spectrometry in combination with H/D exchange. To further confirm the fragments's structures, off-line Fourier transform resonance tandem mass spectrometry (FT-ICR-MS/MS) was also performed. The fragmentation rules of compounds 2a-2d have been summarized and the plausible schemes for the fragmentation pathways were proposed. In this study, one dephosphorylated ion and two phosphorylated ions were observed in ESI-MS(2) spectra of [M+Na](+) ions for compounds 2a-2d. The possible mechanisms about phosphorylation and dephosphorylation were proposed and confirmed by H/D exchange. For the "dephosphorylation" rearrangement, a nitrogen atom was migrated from the phosphoryl group to the carbon atom of Brefeldin A's backbone with losing a molecule of C3H7PO3 (122 Da). For the "phosphorylation" rearrangement, an oxygen atom of one phosphoryl group attacked the sideward phosphorus atom to form a nine-member ring intermediate, then two steps of CH covalent bond cleavage with consecutive migration of hydrogen atom to lose a molecule of C16H20O2 (244 Da). The two proposed rearrangement mechanisms about phosphoryl group transfer might be valuable for the structure analysis of other analogs and provide insights into elucidating the dynamic process of the phosphorylation-dephosphorylation of proteins.

[One case of parotid eosinophilic lymphogranuloma].

The clinical manifestation was painless mass in the parotid gland. Physical examination showed regional swelling in parotid area. Bultrasound examination demonstrated the mass was an hypoechoic nodules of bilateral parotid gland, the border was vague. Absolute value and ratio of peripheral eosinophils were both significantly increased. Pathological examination: parotid eosinophilic lymphogranuloma.

[Investigation of chronic rhinosinusitis on junior middle school students in Zhengzhou area in 2009].

OBJECTIVE: To investigate the incidence of chronic rhinosinusitis and its influence on life quality for junior middle school students in Zhengzhou Municipal. METHODS: Two thousand and twenty junior middle school students in Zhengzhou Municipal were randomly selected as the object of investigation by designed survey and visual analogue scale (VAS) questionnaire of chronic rhinosinusitis the sino-nasal outcome test-20 (SNOT-20) and nasal examination. Effect on the quality of life was investigated based on the degree of troubles caused by symptoms. RESULTS: The incidence of chronic rhinosinusitis in junior middle school students in Zhengzhou Municipal was 6.73% (136/2 020), in which, 22.79% (31/136) of these students' quality of life was affected (VAS>5). CONCLUSIONS: Chronic rhinosinusitis is common in junior middle school students, and the life quality is affected. So we should pay attention to this diseases and take the positive and effective intervention measures.

Cloning, expression and characterization of antimicrobial porcine ß defensin 1 in Escherichia coli.

Porcine ß defensin 1 (pBD1) is a cationic antimicrobial peptide with three pairs of disulfide bonds. When expressed in insect cells, two polypeptides of different length (pBD1(38) and pBD1(42)) accumulated, which differed by N-terminal truncation. However, only pBD1(42) was found in pigs. pBD1(42) had stronger antimicrobial activity than pBD1(38), and thus could be a good candidate as a bactericidal agent for pigs. In this study, pBD1(42) gene, obtained by RT-PCR using the tongue total RNA as a template, was cloned into pET30a expression vector and transformed into Escherichia coli BL21 (DE3) plysS. The recombinant pBD1(42) was expressed after induction by IPTG and purified by His tag affinity column with 90% purity. The recombinant pBD1(42) exhibited antimicrobial activity against both Gram-positive Staphylococcus aureus and Gram-negative E. coli including the multi-resistant E. coli. The minimum inhibitory concentrations (MICs) of recombinant pBD1(42) against tested bacteria were 100 µg/mL for E. coli and 80 µg/mL for S. aureus. In addition, pBD1(42) showed low hemolytic activity and high thermal stability. These properties are relevant for the biotechnological applications of the peptide.

Identification of compatibility between ooplasmic factor and sperm gene in the intersubspecific crosses involving DDK and PWK mice strains.

The DDK strain (Mus musculus domesticus) of inbred mouse has a unique peculiarity known as DDK syndrome. The DDK females are mostly infertile when crossed with males of other inbred strains, while DDK males exhibit normal fertility in the reciprocal crosses, as intrastrain matings. This DDK syndrome has been demonstrated to be caused by an incompatibility system between DDK ooplasmic factor and the sperm gene of other strains owing to the ovum mutant (Om) locus on mouse Chromosome 11. Recently, it was reported that DDK females are fully fertile when crossed to males of MOM (M. m. molossinus) and CASP (M. m. castaneus) strains, indicating that no incompatibilities exist between DDK ooplasmic factor and sperm gene of MOM or CASP males. In the present study, DDK females were found to be also fully fertile when crossed to the males of PWK wild-derived inbred strain (originated from Czech Republic wild mice, M. m. musculus). The crosses of DDK females×F(1) (DDK♀×PWK♂) males also resulted in normal fertility. Furthermore, the transmission ratios of Om alleles from these F(1) males to their backcross N(2) offspring are 50%:50% as genotyped by microsatellite markers closely linked to Om locus. Moreover, it was demonstrated that PWK females are also fully fertile when crossed to DDK males. All above results indicated that no incompatibility exists between ooplasmic factor and sperm gene in the intersubspecific crosses with DDK and PWK strains. PWK strain would also be useful for further investigations on the DDK syndrome, and DDK strain can be used more widely for various studies in the mouse.