Single-cell analysis of graft-infiltrating host cells identifies caspase-1 as a potential therapeutic target for heart transplant rejection.

Aims: Understanding the cellular mechanisms underlying early allograft rejection is crucial for the development of effective immunosuppressant strategies. This study aims to investigate the cellular composition of graft-infiltrating cells during the early rejection stage at a single-cell level and identify potential therapeutic targets. Methods: A heterotopic heart transplant model was established using enhanced green fluorescent protein (eGFP)-expressing mice as recipients of allogeneic or syngeneic grafts. At 3 days post-transplant, eGFP-positive cells infiltrating the grafts were sorted and subjected to single-cell RNA-seq analysis. Potential molecular targets were evaluated by assessing graft survival and functions following administration of various pharmacological inhibitors. Results: A total of 27,053 cells recovered from syngrafts and allografts were classified into 20 clusters based on expression profiles and annotated with a reference dataset. Innate immune cells, including monocytes, macrophages, neutrophils, and dendritic cells, constituted the major infiltrating cell types (>90%) in the grafts. Lymphocytes, fibroblasts, and endothelial cells represented a smaller population. Allografts exhibited significantly increased proportions of monocyte-derived cells involved in antigen processing and presentation, as well as activated lymphocytes, as compared to syngrafts. Differential expression analysis revealed upregulation of interferon activation-related genes in the innate immune cells infiltrating allografts. Pro-inflammatory polarization gene signatures were also enriched in these infiltrating cells of allografts. Gene profiling and intercellular communication analysis identified natural killer cells as the primary source of interferon-γ signaling, activating inflammatory monocytes that displayed strong signals of major histocompatibility complexes and co-stimulatory molecules. The inflammatory response was also associated with promoted T cell proliferation and activation in allografts during the early transplant stages. Notably, caspase-1 exhibited specific upregulation in inflammatory monocytes in response to interferon signaling. The regulon analysis also revealed a significant enrichment of interferon-related motifs within the transcriptional regulatory network of downstream inflammatory genes including caspase-1. Remarkably, pharmacological inhibition of caspase-1 was shown to reduce immune infiltration, prevent acute graft rejection, and improve cardiac contractile function. Conclusion: The single-cell transcriptional profile highlighted the crucial role of caspase-1 in interferon-mediated inflammatory monocytes infiltrating heart transplants, suggesting its potential as a therapeutic target for attenuating rejection.

Epidemiology and Association Rules Analysis for Pulmonary Tuberculosis Cases with Extrapulmonary Tuberculosis from Age and Gender Perspective: A Large-Scale Retrospective Multicenter Observational Study in China.

Background: Tuberculosis (TB), a multisystemic disease with protean presentation, remains a major global health problem. Although concurrent pulmonary tuberculosis (PTB) and extrapulmonary tuberculosis (EPTB) cases are commonly observed clinically, knowledge regarding concurrent PTB-EPTB is limited. Here, a large-scale multicenter observational study conducted in China aimed to study the epidemiology of concurrent PTB-EPTB cases by diagnostically defining TB types and then implementing association rules analysis. Methods: The retrospective study was conducted at 21 hospitals in 15 provinces in China and included all inpatients with confirmed TB diagnoses admitted from Jan 2011 to Dec 2017. Association rules analysis was conducted for cases with concurrent PTB and various types of EPTB using the Apriori algorithm. Results: Evaluation of 438,979TB inpatients indicated PTB was the most commonly diagnosed (82.05%) followed by tuberculous pleurisy (23.62%). Concurrent PTB-EPTB was found in 129,422 cases (29.48%) of which tuberculous pleurisy was the most common concurrent EPTB type observed. The multivariable logistic regression models demonstrated that odds ratios of concurrent PTB-EPTB cases varied by gender and age group. For PTB cases with concurrent EPTB, the strongest association was found between PTB and concurrent bronchial tuberculosis (lift = 1.09). For EPTB cases with concurrent PTB, the strongest association was found between pharyngeal/laryngeal tuberculosis and concurrent PTB (lift = 1.11). Confidence and lift values of concurrent PTB-EPTB cases varied with gender and age. Conclusions: Numerous concurrent PTB-EPTB case types were observed, with confidence and lift values varying with gender and age. Clinicians should screen for concurrent PTB-EPTB in order to improve treatment outcomes.

p18INK4C and BRCA1 inhibit follicular cell proliferation and dedifferentiation in thyroid cancer.

Only 3% of thyroid cancers are medullary thyroid carcinomas (MTCs), the rest are follicular epithelial cell derived non-MTCs (NMTCs). A dysfunctional INK4-CDK4-RB pathway is detected in most of NMTCs. DNA repair defects and genome instability are associated with NMTC dedifferentiation and aggressiveness. Whether inactivation of the INK4-CDK4-RB pathway induces NMTCs and how differentiation of NMTC cells is controlled remain elusive. In this study, we generated p18Ink4c and Brca1 singly and doubly deficient mice as well as p16Ink4a and Brca1 singly and doubly deficient mice. By using these mice and human thyroid carcinoma cell lines, we discovered that loss of p18Ink4c, not p16Ink4a, in mice stimulated follicular cell proliferation and induced NMTCs. Depletion of Brca1 alone or both p16Ink4a and Brca1 did not induce thyroid tumor. Depletion of Brca1 in p18Ink4c null mice results in poorly differentiated and aggressive NMTCs with epithelial-mesenchymal transition (EMT) features and enhanced DNA damage. Knockdown of BRCA1 in thyroid carcinoma cells activated EMT and promoted tumorigenesis whereas overexpression of BRCA1 inhibited EMT. BRCA1 and EMT marker expression were inversely related in human thyroid cancers. Our finding, for the first time, demonstrates that inactivation of INK4-CDK4-RB pathway induces NMTCs and that Brca1 deficiency promotes dedifferentiation of NMTC cells. These results suggest that BRCA1 and p18INK4C collaboratively suppress thyroid tumorigenesis and progression and CDK4 inhibitors will be effective for treatment of INK4-inactivated or cyclin D-overexpressed thyroid carcinomas.

Loss of function of GATA3 induces basal-like mammary tumors.

Purpose: GATA3 is a transcription factor essential for mammary luminal epithelial cell differentiation. Expression of GATA3 is absent or significantly reduced in basal-like breast cancers. Gata3 loss-of-function impairs cell proliferation, making it difficult to investigate the role of GATA3 deficiency in vivo. We previously demonstrated that CDK inhibitor p18INK4c (p18) is a downstream target of GATA3 and restrains mammary epithelial cell proliferation and tumorigenesis. Whether and how loss-of-function of GATA3 results in basal-like breast cancers remains elusive. Methods: We generated mutant mouse strains with heterozygous germline deletion of Gata3 in p18 deficient backgrounds and developed a Gata3 depleted mammary tumor model system to determine the role of Gata3 loss in controlling cell proliferation and aberrant differentiation in mammary tumor development and progression. Results: Haploid loss of Gata3 reduced mammary epithelial cell proliferation with induction of p18, impaired luminal differentiation, and promoted basal differentiation in mammary glands. p18 deficiency induced luminal type mammary tumors and rescued the proliferative defect caused by haploid loss of Gata3. Haploid loss of Gata3 accelerated p18 deficient mammary tumor development and changed the properties of these tumors, resulting in their malignant and luminal-to-basal transformation. Expression of Gata3 negatively correlated with basal differentiation markers in MMTV-PyMT mammary tumor cells. Depletion of Gata3 in luminal tumor cells also reduced cell proliferation with induction of p18 and promoted basal differentiation. We confirmed that expression of GATA3 and basal markers are inversely correlated in human basal-like breast cancers. Conclusions: This study provides the first genetic evidence demonstrating that loss-of-function of GATA3 directly induces basal-like breast cancer. Our finding suggests that basal-like breast cancer may also originate from luminal type cancer.

Epidemiology of concurrent extrapulmonary tuberculosis in inpatients with extrapulmonary tuberculosis lesions in China: a large-scale observational multi-centre investigation.

AIMS: A high proportion of all patients with tuberculosis (TB) present with extrapulmonary TB (EPTB), including concurrent EPTB involving more than one extrapulmonary lesion site. However, previous reports only characterized lesions of single-site EPTB cases. This study aimed to investigate epidemiological characteristics and association rules of concurrent EPTB cases in China. METHODS: An observational multi-centre study of 208,214 patients with EPTB lesions was undertaken in China from January 2011 to December 2017. Multi-variable logistic regression analysis was used to identify associations between gender and concurrent EPTB, and age and concurrent EPTB. Association rules were analysed for significance using the Apriori algorithm. RESULTS: The most common EPTB lesion was tuberculous pleurisy (49.8%), followed by bronchial TB (14.8%) and tuberculous meningitis (7.6%). The most common type of concurrent EPTB was tuberculous pleurisy concurrent with tuberculous peritonitis (1.80%). In total, 22 association rules, including 20 strong association rules, were identified; among these, the highest confidence rates were found for tuberculous myelitis concurrent with tuberculous meningitis, and sacral TB concurrent with lumbar vertebral TB. The association rules of EPTB concurrent with other EPTB types were found to vary with gender and age. The confidence rate of tuberculous myelitis concurrent with tuberculous meningitis was higher in females (83.67%) than males, and was highest in patients aged 25-34 years (87.50%). CONCLUSIONS: Many types of concurrent EPTB were found. Greater awareness of concurrent EPTB disease characteristics is needed to ensure timely clinical diagnosis and treatment of this disease.

The prevalence and risks of major comorbidities among inpatients with pulmonary tuberculosis in China from a gender and age perspective: a large-scale multicenter observational study.

In clinical practice, PTB patients have concurrent many types of comorbidities such as pneumonia, liver disorder, diabetes mellitus, hematological disorder, and malnutrition. Detecting and treating specific comorbidities and preventing their development are important for PTB patients. However, the prevalence of most comorbid conditions in patients with PTB is not well described. We conducted a large-scale, multicenter, observational study to elucidate and illustrate the prevalence rates of major comorbidities in inpatients at 21 hospitals in China. The 19 specific comorbidities were selected for analysis in this patient cohort, and stratified the inpatient cohort according to age and gender. A total of 355,929 PTB inpatients were included, with a male:female ratio of 1.98 and the proportion of ≥ 65 years PTB inpatients was the most. Approximately 70% of PTB inpatients had at least one defined type of comorbidity. The prevalence of 19 specific comorbidities in inpatients with PTB was analyzed, with pneumonia being the most common comorbidity. The prevalence of most comorbidities was higher in males with PTB except thyroid disorders, mental health disorders, etc. The prevalence of defined most comorbidities in patients with PTB tended to increase with increasing age, although some specific comorbidities tended to increase initially then decrease with increasing age. Our study describes multiple clinically important comorbidities among PTB inpatients, and their prevalence between different gender and age groups. The results will enhance the clinical aptitude of physicians who treat patients with PTB to recognize, diagnose, and treat PTB comorbidities early.

The epidemiology of extrapulmonary tuberculosis in China: A large-scale multi-center observational study.

Tuberculosis (TB) remains a serious global public health problem in the present. TB also affects other sites (extrapulmonary tuberculosis, EPTB), and accounts for a significant proportion of tuberculosis cases worldwide. In order to comprehensively understand epidemiology of EBTB in China, and improve early diagnosis and treatment, we conducted a large-scale multi-center observational study to assess the demographic data and the prevalence of common EPTB inpatients, and further evaluate the prevalence of EPTB concurrent with Pulmonary tuberculosis (PTB) and the associations between multiple EPTB types and gender-age group in China. All consecutive age≥15yr inpatients with a confirmed diagnosis of EPTB during the period from January 2011 to December 2017 were included in the study. The descriptive statistical analysis included median and quartile measurements for continuous variables, and frequencies and proportions with 95% confidence intervals (CIs) for categorical variables. Multinomial logistic regression analysis was used to compare the association of multiple EPTB types between age group and gender. The results showed that the proportion of 15-24 years and 25-34 years in EPTB inpatients were the most and the ratio of male: female was 1.51. Approximately 70% of EPTB inpatients were concurrent with PTB or other types of EPTB. The most common of EPTB was tuberculous pleurisy (50.15%), followed by bronchial tuberculosis (14.96%), tuberculous lymphadenitis of the neck (7.24%), tuberculous meningitis (7.23%), etc. It was found that many EPTB inpatients concurrent with PTB. The highest prevalence of EPTB concurrent with PTB was pharyngeal/laryngeal tuberculosis (91.31%), followed by bronchial tuberculosis (89.52%), tuberculosis of hilar lymph nodes (79.52%), tuberculosis of mediastinal lymph nodes (79.13%), intestinal tuberculosis (72.04%), tuberculous pleurisy (65.31%) and tuberculous meningitis (62.64%), etc. The results from EPTB concurrent with PTB suggested that females EPTB inpatients were less likely to be at higher risk of concurrent PTB (aOR = 0.819, 95%CI:0.803-0.835) after adjusted by age. As age increasing, the trend risk of concurrent PTB decreased (aOR = 0.994, 95%CI: 0.989-0.999) after adjusted by gender. Our study demonstrated that the common EPTB were tuberculous pleurisy, bronchial tuberculosis, tuberculous lymphadenitis of the neck, tuberculous meningitis, etc. A majority of patients with pharyngeal/laryngeal tuberculosis, bronchial tuberculosis, tuberculosis of hilar/mediastinal lymph nodes, intestinal tuberculosis, tuberculous pleurisy, tuberculous meningitis, etc. were concurrent with PTB. Female EPTB inpatients were less likely to be at higher risk of concurrent PTB, and as age increasing, the trend risk of concurrent PTB decreased. The clinicians should be alert to the presence of concurrent tuberculosis in EPTB, and all suspected cases of EPTB should be assessed for concomitant PTB to determine whether the case is infectious and to help for early diagnosis and treatment.

Constraints of coal mining safety management efficiency.

BACKGROUND: Investigation of the safety management efficiency in a coal mine aims to improve its safety management level thus ensuring coal mining safety. However, the safety management efficiency is affected by many factors especially for those coal mines operated underground. Furthermore, the constraint factors that are difficult to be identified and eliminated may impede safety management efficiency. OBJECTIVE: This work aims to explore the constraints affecting safety management efficiency through a mathematical model accompanied by some effective measures guided by the theory of constraint (TOC). METHODS: An index system for coal mining safety management efficiency (CMSME) is first established. Then a mathematical model roughly identifying the constraint factors is constructed. The principle of the proposed model is a comparison with the changes of the ratio of integrated CMSME and the ratio of each impact factor over a certain period. Thus, a constraint factor may be one whose ratio changes at a slower rate than that of the integrated CMSME. Following this, some measures are adopted to identify one, or more, real constraints. Finally, the constraints may be broken by internal, or external, means. RESULTS: A case study from Quandian coal mine verified the proposed method: the constraints affecting CMSME could be identified and broken through during the production. This research currently is applied to coal mining activities in a few coal mines, and it will be widely used in the future. CONCLUSIONS: This paper provides a novel method investigating the constraints affecting CMSME and breaking through them. The case study shows that breaking through constraints during the production is beneficial to CMSME. Furthermore, a coal mine with a high CMSME index may still, at some time, have one, or more, bottleneck constraints.

Research status of surgical diagnosis and treatment of chest wall tuberculosis / 中国胸心血管外科临床杂志

@#Chest wall tuberculosis is a common extra-pulmonary tuberculosis, which often occurs in lung or pleura tuberculosis, or coexists with tuberculosis in other parts. The final manifestation of the disease is cold abscess on the chest wall, chronic sinuses with repeated exudation after the abscess is broken. At present, There were a series of problems in the diagnosis, treatment, and extremely prognosis of tuberculosis of the chest wall. Therefore, we reviewed the diagnosis of chest wall tuberculosis, including B-ultrasound, chest CT and positron emission tomography/computed tomography (PET/CT), tuberculous infectin of T cells spot test (T-SPOT TB), pathological examination. The related treatments including medical treatment, surgical treatment and other local treatment in order to better understand the chest wall tuberculosis.

Generalized Grey Target Decision Method for Mixed Attributes Based on Kullback-Leibler Distance.

A novel generalized grey target decision method for mixed attributes based on Kullback-Leibler (K-L) distance is proposed. The proposed approach involves the following steps: first, all indices are converted into index binary connection number vectors; second, the two-tuple (determinacy, uncertainty) numbers originated from index binary connection number vectors are obtained; third, the positive and negative target centers of two-tuple (determinacy, uncertainty) numbers are calculated; then the K-L distances of all alternatives to their positive and negative target centers are integrated by the Technique for Order Preference by Similarity to an Ideal Solution (TOPSIS) method; the final decision is based on the integrated value on a bigger the better basis. A case study exemplifies the proposed approach.

p16INK4a suppresses BRCA1-deficient mammary tumorigenesis.

Senescence prevents the proliferation of genomically damaged, but otherwise replication competent cells at risk of neoplastic transformation. p16INK4A (p16), an inhibitor of CDK4 and CDK6, plays a critical role in controlling cellular senescence in multiple organs. Functional inactivation of p16 by gene mutation and promoter methylation is frequently detected in human breast cancers. However, deleting p16 in mice or targeting DNA methylation within the murine p16 promoter does not result in mammary tumorigenesis. How loss of p16 contributes to mammary tumorigenesis in vivo is not fully understood.In this article, we reported that disruption of Brca1 in the mammary epithelium resulted in premature senescence that was rescued by p16 loss. We found that p16 loss transformed Brca1-deficient mammary epithelial cells and induced mammary tumors, though p16 loss alone was not sufficient to induce mammary tumorigenesis. We demonstrated that loss of both p16 and Brca1 led to metastatic, basal-like, mammary tumors with the induction of EMT and an enrichment of tumor initiating cells. We discovered that promoter methylation silenced p16 expression in most of the tumors developed in mice heterozygous for p16 and lacking Brca1. These data not only identified the function of p16 in suppressing BRCA1-deficient mammary tumorigenesis, but also revealed a collaborative effect of genetic mutation of p16 and epigenetic silencing of its transcription in promoting tumorigenesis. To the best of our knowledge, this is the first genetic evidence directly showing that p16 which is frequently deleted and inactivated in human breast cancers, collaborates with Brca1 controlling mammary tumorigenesis.

Gata3 restrains B cell proliferation and cooperates with p18INK4c to repress B cell lymphomagenesis.

GATA3, a lineage specifier, controls lymphoid cell differentiation and its function in T cell commitment and development has been extensively studied. GATA3 promotes T cell specification by repressing B cell potential in pro T cells and decreased GATA3 expression is essential for early B cell commitment. Inherited genetic variation in GATA3 has been associated with lymphoma susceptibility. However, it remains elusive how the loss of function of GATA3 promotes B cell development and induces B cell lymphomas. In this study, we found that haploid loss of Gata3 by heterozygous germline deletion increased B cell populations in the bone marrow (BM) and spleen, and decreased CD4 T cell populations in the thymus, confirming that Gata3 promotes T and suppresses B cell development. We discovered that haploid loss of Gata3 reduced thymocyte proliferation with induction of p18Ink4c (p18), an inhibitor of CDK4 and CDK6, but enhanced B cell proliferation in the BM and spleen independent of p18. Loss of p18 partially restored Gata3 deficient thymocyte proliferation, but further stimulated Gata3 deficient B cell proliferation in the BM and spleen. Furthermore, we discovered that haploid loss of Gata3 in p18 deficient mice led to the development of B cell lymphomas that were capable of rapidly regenerating tumors when transplanted into immunocompromised mice. These results indicate that Gata3 deficiency promotes B cell differentiation and proliferation, and cooperates with p18 loss to induce B cell lymphomas. This study, for the first time, reveals that Gata3 is a tumor suppressor specifically in B cell lymphomagenesis.

Therapeutic evaluation of video-assisted thoracoscopic surgery versus open thoracotomy for pediatric pulmonary hydatid disease.

BACKGROUND: Hydatid disease is a severe and widespread human cestode infection, and in children, the lung is the most commonly infected organ. In current practice, the standard surgical procedure for the removal of pulmonary hydatid cysts is thoracotomy; therefore, we evaluated the efficacy and safety of video-assisted thoracoscopic surgery (VATS) to treat pediatric pulmonary hydatid disease. To our knowledge, this is the first and large sample comparative study of VATS and thoracotomy for pediatric pulmonary hydatid disease. METHODS: In this study, we retrospectively reviewed 44 (61.1 %) pediatric patients who underwent VATS, and 28 (38.9 %) pediatric patients who underwent conventional thoracotomy from January 2005 to June 2012. Perioperative data, including basic characteristics of patients, the length of hospital stay, intraoperative blood loss, thoracic intubation indwelling time, and complications were compared between VATS and thoracotomy in 72 children with pulmonary hydatid disease. RESULTS: VATS was found to be a safe technique for the treatment of pediatric pulmonary hydatid disease, with zero intraoperative deaths. In the VATS and thoracotomy groups, the hospital stay durations were 10.50 ± 1.20 days and 17.30 ± 2.75 days, respectively, and occurrence rates of complications were 9.1 % (4/44) and 17.9 % (5/28), respectively. The hospital stays were shorter and the hospitalization costs was reduced for the patients who underwent VATS compared with conventional thoracotomy (P = 0.001). Although no statistically significant difference in the recurrence rates (P = 0.958) and complication incidence (P = 0.273) between the two surgical groups was observed, less intraoperative bleeding, shorter thoracic intubation indwelling time and reduced postoperative pain were observed in the patients who underwent VATS (P = 0.001). CONCLUSION: Our study demonstrates the feasibility and safety of VATS for pediatric pulmonary hydatid disease treatment, providing a practice-changing concept for the treatment of this disease in the community. VATS can be a promising therapeutic tool, by overcoming many of the drawbacks of thoracotomy, and can be used as an alternative to thoracotomy for selected pediatric patients.

Application of a bioengineered composite neotrachea in a dog model.

BACKGROUND: Surgical treatment of extensive tracheal lesions remains a major challenge because of lack of an ideal airway substitute that is well vascularized, rigid, and autologous. We describe a novel surgical technique of tracheal reconstruction using a combination of a polypropylene mesh material and anterior cervical myocutaneous flap in a dog model. MATERIALS AND METHODS: A 3.5-4 cm length of cervical trachea was resected in 16 dogs and replaced with a myocutaneous cervical neck flap wrapped around the plain polypropylene tube (group 1, n = 7) or wrapped around a composite of polypropylene tube with an implanted Z-type metallic-covered stent (group 2, n = 9). The cervical tracheal defect was repaired with the previously mentioned substitute that was directly sutured to the remaining tracheal ends. Dogs were followed up using bronchoscopy and x-rays and euthanized at predetermined times for histologic examination. RESULTS: In group 1, four dogs died within 2 wk from respiratory failure with varying degrees of airway collapse and difficulties in expectoration. In group 2, eight dogs survived, whereas one died of anastomotic dehiscence 1 wk after surgery. Necropsy and histologic examination of the anastomotic sites revealed good healing tissue. Pathologic examination also revealed excellent healing of the squamous epithelium of the neotrachea and the columnar epithelium of the native tracheal mucosa. CONCLUSIONS: The tissue compatibility of the polypropylene mesh material and anterior cervical myocutaneous skin flap makes this a promising therapeutic substitute for treatment of patients with extensive tracheal lesions.

[Treatment of 42 child cases of pulmonary echinococcosis granulosus by excision of internal cyst through video-assisted thoracoscopic surgery].

Among 42 child cases of pulmonary echinococcosis granulosus treated by excision of internal cyst through video-assisted thoracoscopic surgery (VATS), the cysts were completely removed in 28 cases (66.7%), cysts in 14 cases (33.3%) were removed by puncture, and cyst ruptured accidentally during operation in 1 cases (2.4%). The mean operation time was (96.70 +/- 10.90) min, mean volume of blood loss during operation was (8.60 +/- 1.31) ml, and the average admission in hospital was (10.20 +/- 1.10) d, with post-operation complications in 2 cases (4.8%). Three years' follow-up revealed no recurrence. The VATS is a way for treating pulmonary echinococcosis granulosus in children with light trauma, less bleeding and fewer complications.