Skin infectome of patients with a tick bite history.

Introduction: Ticks are the most important obligate blood-feeding vectors of human pathogens. With the advance of high-throughput sequencing, more and more bacterial community and virome in tick has been reported, which seems to pose a great threat to people. Methods: A total of 14 skin specimens collected from tick-bite patients with mild to severe symptoms were analyzed through meta-transcriptomic sequencings. Results: Four bacteria genera were both detected in the skins and ticks, including Pseudomonas, Acinetobacter, Corynebacterium and Propionibacterium, and three tick-associated viruses, Jingmen tick virus (JMTV), Bole tick virus 4 (BLTV4) and Deer tick mononegavirales-like virus (DTMV) were identified in the skin samples. Except of known pathogens such as pathogenic rickettsia, Coxiella burnetii and JMTV, we suggest Roseomonas cervicalis and BLTV4 as potential new agents amplified in the skins and then disseminated into the blood. As early as 1 day after a tick-bite, these pathogens can transmit to skins and at most four ones can co-infect in skins. Discussion: Advances in sequencing technologies have revealed that the diversity of tick microbiome and virome goes far beyond our previous understanding. This report not only identifies three new potential pathogens in humans but also shows that the skin barrier is vital in preventing horizontal transmissions of tick-associated bacteria or virus communities to the host. It is the first research on patients' skin infectome after a tick bite and demonstrates that more attention should be paid to the cutaneous response to prevent tick-borne illness.

Metavirome of 31 tick species provides a compendium of 1,801 RNA virus genomes.

The increasing prevalence and expanding distribution of tick-borne viruses globally have raised health concerns, but the full repertoire of the tick virome has not been assessed. We sequenced the meta-transcriptomes of 31 different tick species in the Ixodidae and Argasidae families from across mainland China, and identified 724 RNA viruses with distinctive virome compositions among genera. A total of 1,801 assembled and complete or nearly complete viral genomes revealed an extensive diversity of genome architectures of tick-associated viruses, highlighting ticks as a reservoir of RNA viruses. We examined the phylogenies of different virus families to investigate virome evolution and found that the most diverse tick-associated viruses are positive-strand RNA virus families that demonstrate more ancient divergence than other arboviruses. Tick-specific viruses are often associated with only a few tick species, whereas virus clades that can infect vertebrates are found in a wider range of tick species. We hypothesize that tick viruses can exhibit both 'specialist' and 'generalist' evolutionary trends. We hope that our virome dataset will enable much-needed research on vertebrate-pathogenic tick-associated viruses.

Dermacentor silvarum, a Medically Important Tick, May Not Be a Competent Vector to Transmit Jingmen Tick Virus.

Background: Jingmen tick virus (JMTV) has attracted great attention due to its potential pathogenicity in humans and its transmission by ticks. Dermacentor silvarum (D. silvarum) is one of the dominant tick species in northeastern China, and can transmit many pathogens to humans and animals. However, there have been no report of transmission of JMTV by D. silvarum. Materials and Methods: Ticks were collected from vegetation at the Aershan Port in Inner Mongolia in April 2019. And we do attempt to infect D. silvarum with JMTV by the immersion technique in laboratory conditions. The transmission of JMTV was examined by reverse transcriptase PCR, fluorescence in situ hybridization, and indirect immunofluorescence assay. Statistical analysis was performed using SPSS 24.0. Results: We found that JMTV may only be maintained in the tick without replication, and could not be transmitted to a host following transstadial transmission. Moreover, no virus colonization was found in the midgut or salivary glands of unfed D. silvarum; therefore, D. silvarum may not be susceptible to JMTV infection and therefore unlikely to carry and transmit JMTV. Conclusion: Our study has to some extent filled the knowledge gap regarding the possibility of JMTV transmission by a medically important tick vector, D. silvarum.

Chronic Progression of Lung Cancer Recurrence After Surgery: Warning Role of Postoperative Pneumonia.

PURPOSE: The association between the process of postoperative pneumonia and lung cancer recurrence remains elusive in lung cancer surgery. Herein, the association between postoperative pneumonia and lung cancer recurrence was investigated, emphasizing the warning role of postoperative specific pneumonia in primary lung cancer resection patients. METHODS: The occurrence of postoperative pneumonia was assessed in 4-6 months (PPFS), 7-12 months (PPST), and lung cancer recurrence within 1 year (LRO) in 332 patients. The primary outcome was the development of PPST and LRO according to PPFS occurrence. The relevant risk factors of PPFS, PPST, and LRO were identified through multivariable regression analysis. RESULTS: During follow-up, 151 (45.48%) participants experienced PPFS. Irrespective of the existing postoperative pneumonia in 1-3 months (PPOT), PPFS significantly increased the risk of PPST (P < 0.01) and LRO (P < 0.01), and persistent PPST further increased the risk of LRO (P < 0.001). The generalized estimating equation identified chemotherapy as an independent risk factor for PPFS and PPST. CONCLUSION: PPFS was associated with the increased risk of PPST and LRO. Postoperative pulmonary inflammation assessed 4 months post-surgery also significantly influenced LRO development, indicating a need for close follow-up of lung inflammatory conditions to improve patient outcomes.

Large-Scale Comparative Analyses of Tick Genomes Elucidate Their Genetic Diversity and Vector Capacities.

Among arthropod vectors, ticks transmit the most diverse human and animal pathogens, leading to an increasing number of new challenges worldwide. Here we sequenced and assembled high-quality genomes of six ixodid tick species and further resequenced 678 tick specimens to understand three key aspects of ticks: genetic diversity, population structure, and pathogen distribution. We explored the genetic basis common to ticks, including heme and hemoglobin digestion, iron metabolism, and reactive oxygen species, and unveiled for the first time that genetic structure and pathogen composition in different tick species are mainly shaped by ecological and geographic factors. We further identified species-specific determinants associated with different host ranges, life cycles, and distributions. The findings of this study are an invaluable resource for research and control of ticks and tick-borne diseases.

Acute liver failure associated with Fructus Psoraleae: a case report and literature review.

BACKGROUND: Fructus Psoraleae is the seed of Psoralea corylifolia Linn. Fructus Psoraleae has been shown to be effective in treating some skin diseases, such as vitiligo. As a main ingredient in five types of herbs in the Qubaibabuqi tablet formula, Fructus Psoraleae plays an important role in the treatment of vitiligo. Fructus Psoraleae has potential hepatotoxicity, thus Qubaibabuqi tablets also have potential liver toxicity. CASE PRESENTATION: A 53-year-old woman who was diagnosed with vitiligo in September 2017 was treated with Qubaibabuqi tablets. After approximately 7 months of treatment, the patient developed a severe, diffuse yellow staining of the skin and sclera in March 2018. On admission, she was diagnosed with acute cholestatic hepatitis associated with Fructus Psoraleae. Despite receiving active treatment, her condition rapidly deteriorated and she died 5 days later due to acute liver failure and multiple organ dysfunction. To the best of our knowledge, there are only six reported cases of liver injury associated with Fructus Psoraleae described in the English language literature; however, cases of acute liver failure associated with the use of Fructus Psoraleae have not been described. CONCLUSION: As a main ingredient in the Qubaibabuqi tablet formula, Fructus Psoraleae has potential hepatotoxicity. This potentially fatal adverse effect should be considered when physicians prescribe Qubaibabuqi tablets.

mTORC1 underlies age-related muscle fiber damage and loss by inducing oxidative stress and catabolism.

Aging leads to skeletal muscle atrophy (i.e., sarcopenia), and muscle fiber loss is a critical component of this process. The mechanisms underlying these age-related changes, however, remain unclear. We show here that mTORC1 signaling is activated in a subset of skeletal muscle fibers in aging mouse and human, colocalized with fiber damage. Activation of mTORC1 in TSC1 knockout mouse muscle fibers increases the content of morphologically abnormal mitochondria and causes progressive oxidative stress, fiber damage, and fiber loss over the lifespan. Transcriptomic profiling reveals that mTORC1's activation increases the expression of growth differentiation factors (GDF3, 5, and 15), and of genes involved in mitochondrial oxidative stress and catabolism. We show that increased GDF15 is sufficient to induce oxidative stress and catabolic changes, and that mTORC1 increases the expression of GDF15 via phosphorylation of STAT3. Inhibition of mTORC1 in aging mouse decreases the expression of GDFs and STAT3's phosphorylation in skeletal muscle, reducing oxidative stress and muscle fiber damage and loss. Thus, chronically increased mTORC1 activity contributes to age-related muscle atrophy, and GDF signaling is a proposed mechanism.

The complete mitochondrial genome of Amblyomma geoemydae (Ixodida: Ixodidae).

The complete mitochondrial genome of Amblyomma geoemydae is reported for the first time in this study. Its entire mitogenome is 14,780 bp in length, contained 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and two non-coding regions. The phylogenetic analysis by Maximum-likelihood method show that A. geoemydae and the others of genus Amblyomma are in the same clade, indicating that A. geoemydae belongs to the genus Amblyomma.

Rapid Detection of Ustilaginoidea virens from Rice using Loop-Mediated Isothermal Amplification Assay.

Ustilaginoidea virens is an important fungus that causes rice false smut disease. This disease significantly reduces both grain yield and quality. Various methods have been developed for the detection of U. virens but most of these methods need sophisticated equipment such as a thermal cycler. Here, we present a loop-mediated isothermal amplification (LAMP) assay for the specific detection of U. virens. This assay used a specific region of the UvG-ß1 gene (212-bp region) to design six LAMP primers. The LAMP assay was optimized by the combination of rapidity, simplicity, and high sensitivity for the detection of about 1 pg of target genomic DNA in the reaction whereas, with polymerase chain reaction (PCR), there was no amplification of DNA with concentrations less than 1 ng. Among the genomic DNA of 22 fungus species and two strains of U. virens, only the tube containing the DNA of U. virens changed to yellowish green with SYBR Green I. The color change was indicative of DNA amplification. No DNA was amplified from either the other 22 fungus species or the negative control. Moreover, 20 spikelets and 22 rice seed samples were used for the detection of rice false smut via LAMP. The results were comparable with conventional PCR. We conclude that gene UvG-ß1 coupled with LAMP assay, can be used for the detection and identification of U. virens gene via LAMP.

Tuning the K+ Concentration in the Tunnels of α-MnO2 To Increase the Content of Oxygen Vacancy for Ozone Elimination.

α-MnO2 is a promising material for ozone catalytic decomposition and the oxygen vacancy is often regarded as the active site for ozone adsorption and decomposition. Here, α-MnO2 nanowire with tunable K+ concentration was prepared through a hydrothermal process in KOH solution. High concentration K+ in the tunnel can expand crystal cell and break the charge balance, leading to a lower average oxidation state (AOS) of Mn, which means abundant oxygen vacancy. DFT calculation has also proven that the samples with higher K+ concentration exhibit lower formation energy for oxygen vacancy. Due to the enormous active oxygen vacancies existing in the α-MnO2 nanowire, the lifetime of the catalyst (corresponding to 100% ozone removal rate, 25 °C) is increased from 3 to 15 h. The FT-IR results confirmed that the accumulation of intermediate oxygen species on the catalyst surface is the main reason why it is deactivated after long time reaction. In this work, the performance of the catalyst has been improved because the abundant active oxygen vacancies are fabricated by the electrostatic interaction between oxygen atoms inside the tunnels and the introduced K+, which offers us a new perspective to design a high efficiency catalyst and may promote manganese oxide for practical ozone elimination.

[Application of small intestine double stoma and succus entericus reinfusion in the patients with severe intra-abdominal infection].

OBJECTIVE: To evaluate the application of small intestine double stoma and succus entericus reinfusion in the patients with severe intra-abdominal infection. METHODS: Ten patients with high intestinal perforation from February 2005 to November 2014 were enrolled in the study. All the cases received emergency operation. Small bowel with intestinal perforation was resected, and double stoma was applied in the proximal and distal small intestine. When abdominal infection under control, total enteral nutrition was successfully administered from nasogastric tube. The succus entericus from the proximal intestine was collected and transfused back to the distal intestine. Stool was collected and fecal nitrogen, fat and carbohydrate contents were determined. Related serum protein levels were measured. RESULTS: As compared to pre-reinfusion, the absorption rate of carbohydrate [(90.9±7.8)% vs. (82.7±15.2)%], fat [(87.6±6.4)% vs. (59.1±10.8)%], and nitrogen [(82.4±9.8)% vs. (67.2±15.4)%] increased after succus entericus reinfusion (P<0.05). The serum protein levels increased significantly as well[fibronectin: (285.6±3.6) vs. (157.0±22.6) mg/L, P<0.01; transferrin: (4.86±0.21) vs. (3.60±0.25) g/L, P<0.05; pre-albumin: (291.3±112.5) vs. (199.1±53.3) mg/L, P<0.05]. CONCLUSION: Small intestine double stoma and succus entericus reinfusion are effective in improving the absorption of carbohydrate, fat and nitrogen in the patients with severe intra-abdominal infection.

Simvastatin combined with aspirin increases the survival time of heart allograft by activating CD4(+)CD25(+) Treg cells and enhancing vascular endothelial cell protection.

OBJECTIVE: The objective was to investigate whether the combination of simvastatin and aspirin treatment prolongs the survival time of the heart allograft in rat and its underlying mechanism. METHODS: Heterotopic heart transplantation was performed using Wistar rats as donors and Sprague-Dawley (SD) rats as recipients. The SD rats were randomly divided into five groups (n=20/group): sham, HT (heart transplantation), HT+simvastatin (HT+S), HT+aspirin (HT+A), and HT+aspirin+simvastatin (HT+A+S). After transplantation, at 3, 7, 10, 15, 20, 30, and 40 days, the endothelial nitric oxide synthase (eNOS) expression was assessed by immunohistological staining; nitric oxide (NO) levels were analyzed by Griess assay; the activation of CD4(+)CD25(+) T regulatory lymphocytes (Tregs) was analyzed by flow cytometry; and pathological changes in the graft heart were determined by histology. RESULTS: Combined treatment of hearts with simvastatin and aspirin significantly prolonged the mean survival time of heart allografts [8 ± 1.2 days (n=18), 20 ± 3.4 days (n=19), 21 ± 2.8 days (n=19), and 39 ± 5.3 days (n=19) for HT, HT+S, HT+A, and HT+A+S group, respectively; HT vs. HT+A+S, P<.001; HT vs. HT+S or HT+A, P<.05]. In addition, CONCLUSIONS: Simvastatin given in combination with aspirin delayed the development of pathological changes in the myocardium, reduced vascular damage and prolonged the survival time of cardiac allograft. The underlying mechanism is linked with CD4(+)CD25(+)-Treg-cell-induced immune tolerance and enhanced vascular endothelial cell protection.

Pentraxin 3, long expression in mononuclear cells of patients with acute coronary syndrome: Correlation with C-reactive protein and matrix metalloproteinase-9 levels.

OBJECTIVES: To investigate expression of pentraxin 3, long (PTX3) in patients with acute coronary syndrome (ACS) and its correlation with matrix metalloproteinase-9 (MMP-9) and C-reactive protein (CRP) levels. METHODS: Patients with ACS were randomly assigned to the ACS group (subdivided into unstable angina pectoris [UAP] and acute myocardial infarction [AMI]). Healthy participants and patients with stable angina pectoris (SAP) were enrolled as controls. Mononuclear cell PTX3 expression, and serum MMP-9 and CRP levels, were measured by enzyme-linked immunosorbent assay. RESULTS: The ACS group comprised 200 patients (80 in the UAP subgroup; 120 in the AMI subgroup). The control group comprised 130 participants (80 healthy volunteers and 50 patients with SAP). PTX3 expression was significantly higher in the ACS group compared with controls (3.64 ± 0.49 versus 1.85 ± 0.65 ng/ml), and significantly higher in the AMI compared with the UAP subgroup (5.44 ± 0.47 versus 3.39 ± 0.59 ng/ml). Serum MMP-9 and CRP levels were significantly higher in the ACS group compared with controls (48.55 ± 14.22 versus 23.14 ± 0.62 ng/ml; 4.88 ± 1.76 versus 1.26 ± 0.19 ng/ml, respectively), and significantly higher in the AMI compared with the UAP subgroup (58.13 ± 7.24 versus 31.77 ± 3.61 ng/ml; 5.80 ± 1.46 versus 3.27 ± 0.83 ng/ml, respectively). PTX3 expression, and MMP-9 and CRP levels in the SAP subgroup, were not significantly different from the healthy participants. PTX3 expression positively correlated with MMP-9 and CRP levels. CONCLUSIONS: In patients with ACS, peripheral blood mononuclear cell PTX3 expression, and serum MMP-9 and CRP levels, were significantly enhanced compared with controls; in addition, PTX3 expression positively correlated with MMP-9 and CRP levels. PTX3 may be involved in ACS pathogenesis.

Splenectomy increases the survival time of heart allograft via developing immune tolerance.

BACKGROUND: The spleen is an active lymphoid organ. The effect of splenectomy on the immune response remains unclear. This study investigated whether splenectomy can induce immune tolerance and has a beneficial role in cardiac allograft. METHODS: Wistar rats were used for heart donors. The Sprague-Dawley (SD) rats designated as the recipients of heart transplantation (HT) were randomly assigned into four groups: sham, splenectomy, HT, splenectomy + HT. The survival of transplanted hearts was assessed by daily checking of abdominal palpation. At various time points after transplantation, the transplanted hearts were collected and histologically examined; the level of CD4+CD25+ T regulatory lymphocytes (Tregs) and rate of lymphocyte apoptosis (annexin-v+ PI+ cells) in the blood were analyzed by using flow cytometric method. RESULTS: 1) Splenectomy significantly prolonged the mean survival time of heart allografts (7 ± 1.1 days and 27 ± 1.5 days for HT and splenectomy + HT, respectively; n = 12-14/group, HT vs. splenectomy + HT, p < 0.001); 2) Splenectomy delayed pathological changes (inflammatory cell infiltration, myocardial damage) of the transplanted hearts in splenectomy + HT rats; 3) The level of CD4+CD25+ Tregs in the blood of splenectomized rats was significantly increased within 7 days (2.4 ± 0.5%, 4.9 ± 1.3% and 5.3 ± 1.0% for sham, splenectomy and splenectomy + HT, respectively; n = 15/group, sham vs. splenectomy or splenectomy + HT, p < 0.05) after splenectomy surgery and gradually decreased to baseline level; 4) Splenectomy increased the rate of lymphocyte apoptosis (day 7: 0.3 ± 0.05%, 3.9 ± 0.9% and 4.1 ± 0.9% for sham, splenectomy and splenectomy + HT, respectively; n = 15/group, sham vs. splenectomy or splenectomy + HT, p < 0.05) in a pattern similar to the change of the CD4+CD25+ Tregs in the blood. CONCLUSIONS: Splenectomy inhibits the development of pathology and prolongs the survival time of cardiac allograft. The responsible mechanism is associated with induction of immune tolerance via elevating CD4+CD25+ Tregs and increasing lymphocyte apoptosis.

[Primary cultured neonate rat sinus node cells: morphology and expression of kir2.1 protein].

OBJECTIVE: To investigate the expression of kir2.1 protein in primary cultured sinus node cells and establish a reliable technique to locate, culture and characterize neonatal rat sinus node cells. METHODS: In paraffin sections, the location and morphology of the neonatal rat sinus node cells were observed by HE staining, silver nitrate staining, myelin staining and phosphotungstic acid-hematoxylin (PTAH) staining. Primary cell culture from the neonatal rat sinus node was conducted to observe the spontaneous contraction frequency, cell morphology and kir2.1 protein expression. RESULTS: Combination of the 3 staining methods allowed accurate localization of the sino-atrial nodal (SAN) tissue, and among the cultured cells in the SAN, at least 3 distinct types of cells with spontaneous contraction were observed. The majority of the contracting cells were spindle cells and their construction and impulse frequency indicated their identity as pacemaker cells, while the triangular and irregular cells resembled the atrial muscle cells. A lower expression level of kir2.1 protein was detected in SAN cells than in the atrial and ventricular myocytes of the neonatal rats. CONCLUSION: Combination of silver nitrate staining, myelin staining and PTAH staining identifies the exact location of the sinus node tissue, and cultured sinus node cells have lower expression of kir2.1 protein than the atrial and ventricular myocytes of neonatal rats.

[FHIT protein expression during malignant transformation of human bronchial epithelial cells induced by NNK].

BACKGROUND: Most studies about FHIT protein expression were performed in normal tracheal epithelium, precancerous lesions and lung cancer tissues respectively, but not in the course of malignant transformation of lung cancer. The aim of this study is to detect the changes of FHIT protein expression during malignant transformation of immortalized human bronchial epithelial cells (BEAS-2B) induced by tobacco-specific nitrosamine (NNK), and to explore its significance. METHODS: BEAS-2B cells were induced to malignantly transform (BEAS-2B NNK) by 500mg/L NNK, and FHIT protein expression was detected in the different passages of BEAS-2B NNK and BEAS-2B cells by SP immunocytochemistry. RESULTS: Part 1: Model of malignant transformation of BEAS-2B cells induced by NNK was established. (1) The serum resistance was significantly increased in the 5th passage of BEAS-2B NNK cells. (2) The anchorage independent growth (soft agar colony formation) appeared in the 15th passage of BEAS-2B NNK cells. (3) The ultrastructure of the 20th passage of BEAS-2B NNK cells showed obvious heteromorphy characterization. (4) The 25th passage of BEAS-2B NNK cells developed into tumors in nude mice, which were well differentiated squamous cell carcinoma. Part 2: FHIT protein was steadily expressed in the different passages of BEAS-2B cells (P > 0.05). FHIT protein expression was obviously decreased from 5th to 15th passage of BEAS-2B NNK cells, but it was unexpectedly overexpressed in the 25th passage. CONCLUSIONS: (1) The model of malignant transformation of BEAS-2B cells induced by NNK (500mg/L) can be established successfully and may be used for investigation of molecular biological mechanisms of lung cancer, especially for smoking-related cases. (2) Decrease of FHIT protein expression might be an early event, however, its overexpression in the late passages should be further studied.