[Analysis of the factors influencing the severity of coronavirus disease 2019 in patients with myeloproliferative neoplasms based on an online questionnaire].

Objective: To explore the variables associated with the severity of coronavirus disease 2019 (COVID-19) caused by the SARS-CoV-2 omicron variant during the epidemic in patients with myeloproliferative neoplasms (MPN). Methods: A cross-sectional study. During the SARS-CoV-2 omicron variant pandemic from December 15, 2022, to March 15, 2023, COVID-19 related data for patients with MPN who were treated at Peking University People's Hospital were collected through an online questionnaire-based survey. All questionnaires and clinical data were checked by medical assistants. Logistic multivariate analysis was used to explore the prevalence and variables associated with the severity of COVID-19 in patients with MPN. Results: A total of 239 patients with MPN, including 90 (37.7%) presenting with essential thrombocythemia (ET), 50 (20.9%) with polycythemia vera (PV), and 99 (41.4%) with myelofibrosis (MF), were enrolled in the study. The 99 patients with MF included 87 (87.9%) with primary MF, 5 (5.1%) with post-PV MF, and 7 (7.1%) with post-ET MF. Overall, 239 (100%) patients reported that they experienced COVID-19 during the pandemic. Of these, 226 (94.6%) had mild disease, 4 (1.7%) had moderate disease, 7 (2.9%) had severe disease, and 2 (0.8%) had critical disease. Two (0.8%) patients with severe COVID-19 died, one of which suffered from MT and the other from PV. Multivariate analysis showed that older age (OR=2.36, 95%CI 1.24-4.49), MF (OR=10.22, 95%CI 1.13-92.80), or comorbidity (OR=5.25, 95%CI 1.25-22.03) were associated with a significantly higher risk of developing moderate, severe, or critical COVID-19. Among patients with MF, higher risk stratification reflected an increased risk of developing moderate, severe, or critical COVID-19 (P=0.034). Conclusion: During the omicron pandemic, older age, MF (especially higher-risk categories), and comorbidity were associated with a higher risk of developing moderate, severe, or critical COVID-19.

Assessing the construction of a Healthy City in China: a conceptual framework and evaluation index system.

OBJECTIVES: COVID-19 has brought challenges to the health of all mankind. It is particularly important to promote the construction of a 'Healthy China' and build a 'healthy community'. The aims of this study were to construct a reasonable conceptual framework for the Healthy City concept and to assess Healthy City construction in China. STUDY DESIGN: This study combined qualitative and quantitative research. METHODS: This study proposes the concept model of 'nature-human body-Healthy City' and accordingly constructs an evaluation index system for the construction of a Healthy City that integrates five dimensions, namely, the medical level, economic basis, cultural development, social services, and ecological environment to explore the spatial and temporal heterogeneity of Healthy City construction in China. Finally, the influencing factors of Healthy City construction patterns are explored using GeoDetector. RESULTS: (1) The pace of Healthy City construction is generally on the rise; (2) the construction of Healthy Cities exhibits significant global spatial autocorrelation and gradually increasing agglomeration. The spatial distribution of cold hotspot areas was relatively stable; (3) medical and health progress is an important factor; the level of economic development is the leading support; the endowment of resources and environment is the basic condition; public service support provides important support; and scientific and technological innovation capabilities provide technical support for the construction of a Healthy City. CONCLUSIONS: The spatial heterogeneity of Healthy City construction in China is evident, and the state of spatial distribution is relatively stable. The spatial pattern of Healthy City construction is shaped by a combination of factors. Our research will provide a scientific basis for promoting the construction of Healthy Cities and helping to implement the Health China Strategy.

[Proteomic study on the damage of learning and memory ability of rat offspring caused by chronic stress during pregnancy].

Objective: To study the differential protein and signal pathway related to the impairment of learning and memory ability of offspring caused by chronic stress during pregnancy and explore the possible mechanism. Methods: From July to October 2019, sixteen SPF free female SD rats aged 80-90 days, weighing (200±20) g. Twelve SPF grade male SD rats aged 90-100 days, weighing (220±20) g. After a week of adaptive feeding, the female rats were randomly divided into control group and model group (8 rats in each group) , male rats were divided into control mating group (n=8) and model mating group (n=4) . Chronic unpredictable mild stress (CUMS) model was established and stimulated continuously for 21 days. One day before stress, the first, seventh, fourteenth and 21th day after stress, the blood was collected from the inner canthus vein of the female rats, and the content of corticosterone was determined. Morris water maze test was used to detect the spatial learning and memory ability of offspring rats. The morphological changes of hippocampus were observed by HE and Nissl staining. The proteomic correlation analysis of offspring rats' hippocampus was performed by isobaric tags for relative and absolute quantification (iTRAQ) technique. Results: Compared with the control group, the content of plasma corticosterone in the model group was significantly higher (F=7.717, P<0.05) , and the model was successfully established. In Morris water maze test, compared with the control offspring group, the escape latency was longer, the average swimming speed was lower, the number of crossing platform was less, and the target quadrant run was shorter in the model offspring group (P<0.05) . The pathological results showed that the morphology of cells in the hippocampal tissue of the model offspring group was irregular, the number of neurons was small, Nissl body was unevenly distributed, the volume was small and the number was small. Mass spectrometry analysis showed that a total of 5065 proteins were screened out in the two offspring groups, and 26 proteins were differentially expressed (P<0.05) , of which 19 proteins were up-regulated and 7 proteins were down regulated. The differential proteins were mainly involved in 23 biological processes, 14 cellular components and 9 molecular functions. Kyoto Encyclopedia of genes and genomes (KEGG) enrichment analysis showed that 57 pathways were enriched, of which 8 signaling pathways were significantly enriched (P<0.05) . There were 5 signaling pathways that might be involved in the impairment of learning and memory ability of offspring, including neuroactive ligand receptor interaction, cGMP-PKG signaling pathway, adhesion and connection, adhesion and connection FoxO signaling pathway and Notch signaling pathway, mainly including tyrosine protein kinase receptor, tyrosine kinase receptor and Notch signaling pathway, and α2A adrenergic receptor, cGMP dependent protein kinase and other differential proteins may be involved in the injury process. Conclusion: The damage of learning and memory ability of offspring may be caused by chronic stress during pregnancy rats. The enriched signal pathway and key differential proteins of proteomics may play an important role in the process of damage.

[Effects of chronic stress during pregnancy on composition and diversity of intestinal microbiota in female rats and offspring].

Objective: To investigate the effect of chronic stress of pregnant rats on the gut microbiota of female rats and offspring, and explore the role of intestinal microbiota in chronic stress during pregnancy. Methods: In November 2019, SPF-grade healthy adult SD rats were selected. 16 female rats were randomly divided into control group and model group, with 8 in each group; 12 male rats were randomly divided into model mating group (8) and control mating group (4) . A model of chronic unpredictable mild stress (CUMS) during pregnancy was established. Blood samples were collected from the iliac vein of the female rats 1 day before and 1, 7, and 14 days after the CUMS protocol, and measured for plasma corticosterone content by radioimmunoassay. After the stress was completed, fresh feces of the female rats were collected for testing. The offspring's fresh stool samples were collected on postnatal day 20 (PND20) , and they were divided into control offspring group and model offspring group samples. The sequence of 16S rRNAV3-V4 regions of microorganisms in the feces of offspring was determined by Illumina MiSeq technique; and the interaction between microbial community structure and diversity were analyzed. Results: The content of plasma corticosterone in the model group was higher than that in the control group on the 7th and 14th day of stress (P<0.05) . Compared with the control group, the Sobs index, Chao index, ACE index and Shannon index of the model group were decreased (P<0.05) . The number of unique species abundance (OTU) in the control group was 130, and 91 in the model group. The relative abundance of female Firmicutes in the control group (64.87%) was higher than that in the model group, and the relative abundance of Bacteroides (31.72%) was lower than that of the model group (46.35%) . The Sobs index, Chao index, ACE index, Simpson index and Shannon index of the control offspring group were higher than those of the model offspring group (P<0.05) . The number of unique OTUs in the model offspring group was 75, and 93 in the control offspring group. The relative abundance of Firmicutes (60.24%) in the control offspring group was higher than that of the model offspring group (52.95%) . Conclusion: Chronic stress during pregnancy can not only lead to the disorder of intestinal flora in female rats, but also lead to the change of intrauterine environment, thus affecting the diversity of intestinal flora in offspring.

[Research on feasibility of in vitro inflammatory wound microenvironment simulated by using inflammatory wound tissue homogenate of mice].

Objective: To investigate the feasibility of in vitro inflammatory wound microenvironment simulated by using inflammatory wound tissue homogenate of mice. Methods: (1) Ten eight-week-old C57BL/6 male mice were collected and full-thickness skin tissue with diameter of 1.0 cm on both sides of the midline of the back was taken with a perforator to make the normal skin tissue homogenate supernatant. At 48 h after the full-thickness skin defect wound was established, the wound tissue within 2 mm from the wound edge was taken to make inflammatory wound tissue homogenate supernatant. Two kinds of tissue homogenate supernatant were taken to adjust the total protein concentration to 1 mg/mL, and the tumor necrosis factor α (TNF-α) content was detected by enzyme-linked immunosorbent assay. The number of sample was 6. (2) The primary passage of human umbilical cord mesenchymal stem cells (hUCMSCs) were collected and cultured to the 3rd passage with the normal exosomes being extracted from the hUCMSCs after cultured for 48 h. Another batch of hUCMSCs in the 3rd passage was collected and stimulated with inflammatory wound tissue homogenate supernatant of 30, 50, and 100 µg/mL total protein and normal skin tissue homogenate supernatant of 30, 50, and 100 µg/mL total protein, respectively. After cultured for 48 h, the exosomes stimulated with normal protein of 30, 50, and 100 µg/mL and exosomes stimulated with inflammatory protein of 30, 50, and 100 µg/mL were extracted. Normal exosomes, exosomes stimulated with 30 µg/mL normal protein, and exosomes stimulated with 30 µg/mL inflammatory protein were collected, the morphology was observed by transmission electron microscope, the particle size was detected by nanoparticle tracking analyzer, and the expressions of CD9 and CD63 were detected by Western blotting. (3) Twenty one-day-old C57BL/6 mice were taken to isolate the primary passage of fibroblasts (Fbs) and the 3rd passage of Fbs, whose morphology was observed under the inverted phase contrast microscope. The Fbs of 3rd passage were collected to observe the expression of vimentin by cell crawling method combined with immunofluorescence method at culture hour (CH) 2. (4) The Fbs of 3rd passage were divided into control group, normal exosome group, 30, 50, 100 µg/mL normal protein stimulating exosome group, and 30, 50, 100 µg/mL inflammatory protein stimulating exosome group according to the random number table, with 4 wells in each group. Cells in control group received no treatment, and cells in the other 7 groups were respectively added with normal exosomes, exosomes stimulated with normal protein of 30, 50, and 100 µg/mL, and exosomes stimulated with inflammatory protein of 30, 50, and 100 µg/mL prepared in experiment (2). The final mass concentration of exosomes was adjusted to 10 µg/mL. The cell viability was detected by cell count kit 8 at CH 48. (5) Two batches of Fbs in the 3rd passage were divided and treated as those in experiment (4), with 4 wells in each group, and the final mass concentration of exosomes was adjusted to 1 and 10 µg/mL, respectively. The cell mobility was detected by cell scratch test at CH 6, 12, and 24. (6) Two batches of the Fbs of 3rd passage were collected, divided, and treated as those in experiment (4) except with no control group, with 3 wells in each group, and the final mass concentration of exosomes was respectively adjusted to 1 and 10 µg/mL. The mRNA expression levels of transforming growth factor ß(1) (TGF-ß(1)), TGF-ß(3), and α smooth muscle actin (α-SMA) were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction at CH 48. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and Bonferroni method. Results: (1) The content of TNF-α in inflammatory wound tissue homogenate supernatant of mice was (116±3) pg/mL, significantly higher than (97±5) pg/mL in normal skin tissue homogenate supernatant at post injury hour 48 (t=3.306, P<0.05). (2) Normal exosomes, exosomes stimulated with 30 µg/mL normal protein, and exosomes stimulated with 30 µg/mL inflammatory protein of hUCMSCs showed the typical saucer-like shape. The particle sizes of the three exosomes of hUCMSCs were 30-150 nm, which were all within the normal particle size range of exosome. Three exosomes of hUCMSCs positively expressed CD9 and CD63. (3) The primary passage of cells were clearly defined and showed protruding spindle shape, irregular polygon shape, or slender strip shape. The morphology of the 3rd and the primary passage of cells is similar. At CH 2, vimentin in cells was positively expressed, and the cells were identified as Fbs. (4) At CH 48, the cell viability was (137.4±2.8)% in 30 µg/mL inflammatory protein stimulating exosome group, obviously higher than 100%, (107.5±2.4)%, (113.3±3.2)%, (104.0±2.0)%, and (101.9±1.5)% in control group, normal exosome group, 30 µg/mL normal protein stimulating exosome group, and 50 and 100 µg/mL inflammatory protein stimulating exosome groups, respectively (P<0.01), and cell viability in 30 µg/mL normal protein stimulating exosome group was obviously higher than that in control group, normal exosome group, and 50 and 100 µg/mL normal protein stimulating exosome groups [(103.4±2.2)% and (102.5±1.4)%], respectively (P<0.01). (5) At CH 6, 12, and 24, the mobility rate of cells in 30 µg/mL inflammatory protein stimulating exosome group was significantly higher than that in control group, normal exosome group, 30 µg/mL normal protein stimulating exosome group, and 50 and 100 µg/mL inflammatory protein stimulating exosome groups, respectively, when the final mass concentrations of exosome was 1 µg/mL (P<0.05) . At CH 12, the mobility rate of cells in 30 µg/mL normal protein stimulating exosome group was obviously higher than that in control group, normal exosome group, and 50 and 100 µg/mL normal protein stimulating exosome groups, respectively, when the final mass concentration of exosome was 1 µg/mL (P<0.05). At CH 6, the mobility rate of cells in 30 µg/mL inflammatory protein stimulating exosome group was significantly higher than that in control group and normal exosome group (P<0.05), and the mobility rate of cells in 30 µg/mL normal protein stimulating exosome group was significantly higher than that in 50 and 100 µg/mL normal protein stimulating exosome groups, respectively, when the final mass concentration of exosome was 10 µg/mL (P<0.05). At CH 12 and 24, the mobility rate of cells in 30 µg/mL inflammatory protein stimulating exosome group was significantly higher than that in control group, normal exosome group, and 50 and 100 µg/mL inflammatory protein stimulating exosome groups (P<0.05), and the mobility rate of cells in 30 µg/mL normal protein stimulating exosome group was significantly higher than that in control group, normal exosome group, and 50 and 100 µg/mL normal protein stimulating exosome groups, respectively, when the final mass concentration of exosome was 10 µg/mL (P<0.05). (6) There were no statistically significant differences in mRNA expression levels of TGF-ß(1), TGF-ß(3), and α-SMA of cells among the 7 groups at CH 48 when the final mass concentration of exosome was 1 µg/mL (F=1.123, 1.537, 1.653, P>0.05). There were no statistically significant differences in mRNA expression levels of TGF-ß(1) and α-SMA of cells among the 7 groups at CH 48 when the final mass concentration of exosome was 10 µg/mL (F=1.487, 1.308, P>0.05), and mRNA expression level of TGF-ß(3) of cells in 50 µg/mL inflammatory protein stimulating exosome group at CH 48 was significantly higher than that in normal exosome group, 50 µg/mL normal protein stimulating exosome group, and 30 and 100 µg/mL inflammatory protein stimulating exosome groups when the final mass concentration of exosome was 10 µg/mL (P<0.05). Conclusions: The pretreatment with inflammatory wound tissue homogenate supernatant of mice has no significant effect on the total protein of hUCMSCs exosomes. The hUCMSCs exosomes stimulated by low concentration inflammatory wound tissue homogenate supernatant can significantly promote the proliferation and migration ability of Fbs. The content of inflammatory mediators in the wound tissue homogenate supernatant during the inflammatory phase is extremely low, which may be the reason that the anti-inflammation and tissue repair paracrine effects of mesenchymal stem cell cannot be effectively started.

MiR-204 inhibits inflammation and cell apoptosis in retinopathy rats with diabetic retinopathy by regulating Bcl-2 and SIRT1 expressions.

OBJECTIVE: To explore the influences of micro ribonucleic acid (miR)-204 on the rats with diabetic retinopathy by regulating the expressions of B-cell lymphoma 2 (Bcl-2) and sirtuin 1 (SIRT1). MATERIALS AND METHODS: A total of 36 Sprague-Dawley rats were randomly assigned into normal group (n=12), model group (n=12), and miR-204 mimics group (n=12). No treatment was performed in the normal group, the diabetic retinopathy model was established in model group, and miR-204 mimics were administered for intervention after modeling in the inhibitor group. After 7 d, materials were sampled for detection. The expressions of Bcl-2 and SIRT1 were detected via immunohistochemistry, and their relative protein expression levels were determined via Western blotting (WB). Quantitative Polymerase Chain Reaction (qPCR) was performed to detect the expression of miR-204, and the content of inflammatory factors interleukin (IL)-6, IL-18, and tumor necrosis factor-α (TNF-α) was measured using enzyme-linked immunosorbent assay (ELISA). Finally, cell apoptosis was evaluated via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). RESULTS: Immunohistochemistry results showed that the positive expression levels of Bcl-2 and SIRT1 were substantially lower in the model and miR-204 mimics groups than those in the normal group (p<0.05), and their positive expression levels in miR-204 mimics group were notably higher than those in model group (p<0.05). According to Western blot (WB) results, the relative protein expression levels of Bcl-2 and SIRT1 markedly declined in the other two groups compared with those in the normal group (p<0.05), while miR-204 mimics group exhibited remarkably higher relative protein expression levels of Bcl-2 and SIRT1 than the model group (p<0.05). The results of qPCR revealed that the relative expression level of miR-204 was markedly lowered in model and miR-204 mimics groups compared with that in the normal group (p<0.05), and its relative expression level in miR-204 mimics group was remarkably higher than that in the model group. It was found through enzyme-linked immunosorbent assay (ELISA) that compared with normal group, the other two groups had substantially increased content of IL-6, IL-18, and TNF-α (p<0.05), and the content of IL-6, IL-18, and TNF-α in miR-204 mimics group was markedly lower than that in the model group (p<0.05). According to TUNEL results, the apoptosis rate of cells rose substantially in the other two groups compared with that in the normal group (p<0.05), while was notably lower in the miR-204 mimics group than that in the model group (p<0.05). CONCLUSIONS: MiR-204 up-regulates Bcl-2 and SIRT1 expressions to inhibit the inflammation and cell apoptosis in rats with diabetic retinopathy.

CRISPR-dCas9-mediated knockdown of prtR, an essential gene in Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a widely distributed non-fermentative Gram-negative opportunistic pathogen that is often responsible for nosocomial infections. Gene interference is a potentially valuable tool for investigating essential genes in P. aeruginosa. To establish a gene interference platform in P. aeruginosa, CRISPR system was used with an inactive Cas9 protein. The CRISPR-dCas9 system was cloned into pHERD20T, a shuttle vector with arabinose inducible promoter, and was further modified to target a regulatory gene prtR that is essential for the viability of P. aeruginosa. Cells expressing the prtR-targeting CRISPR interference (CRISPRi) showed growth defect in an arabinose dose-dependent manner. A high-throughput RNA sequencing analysis of bacterial cells with or without the CRISPRi-mediated prtR inhibition indicated that prtRis a global regulator affecting multiple biological processes. In conclusion, the CRISPR-dCas9-based gene knockdown system has been successfully implemented in P. aeruginosa and demonstrated to be an effective tool in the investigation of essential or difficult-to-inactivate genes in this species.

[Clinical analysis of kidney injury in patients with COVID-19].

Objective: To investigate the relationship between novel coronavirus pneumonia (COVID-19) and kidney injury. Methods: A retrospective analysis was performed on confirmed COVID-19 patients in the Central Theater Command General Hospital of Chinese PLA on March 12, 2020. A total of 87 hospitalized confirmed COVID-19 patients were enrolled in the study, and they were hospitalized for at least one week. The recorded information included clinical data and indicators of kidney-related laboratory tests. Results: The average age of patients was (65.2±17.1) years, and 34.5% (30/87) patients were ≥ 75 years old and 31.0% (27/87) patients were 60-74 years old. Male and female patients accounted for 59.8% (52/87) and 40.2% (35/87), respectively. There were 29.9% (26/87) and 12.6% (11/87) patients who had already showed mild elevation of blood urea nitrogen (BUN) and serum creatinine (SCr) at admission. Moreover, 25.3% (22/87) and 4.6% (4/87) patients still exhibited mild elevation of BUN and SCr one week after admission. However, 28.7% (25/87) patients showed an elevation of BUN one week later after admission, though their BUN levels were normal at admission. Likewise, 16.1% (14/87) patients showed an elevation of SCr one week later after admission, while their SCr levels were normal at admission. Only two patients had an increase of SCr ≥26.5 µmol/L, and both of them were over 75 years old. Conclusions: COVID-19 patients with severe acute kidney injury are uncommon. However, attention should be paid to acute kidney injury of the elderly patients in the diagnosis and treatment of COVID-19.

[Analysis on the situation of being supported by the national natural science foundation of China in the field of occupational diseases from 2010 to 2019].

Objective: To understand the situation supported by the National Natural Science Foundation in the field of occupational diseases (H2402) in China, so as to provide a reference basis for the application and research of scientific researchers in the field of occupational diseases in China. Methods: The information system of scientific and technological achievements was used to search the financial support of the National Natural Science Foundation of China (NSFC) in the field of occupational diseases from 2010 to 2019. Results: From 2010 to 2019, a total of 55 projects were funded under the Occupational Disease code (H2402) , with a total funding of 22.33 million yuan, of which 30 were supported by the Youth Science Foundation, 20 by the Youth Science Foundation and 5 by the Regional Science Foundation. Thirty five items of the research projects focused on pneumoconiosis and other respiratory diseases which accounted for 63.64 per cent. Forty one items of scientific research projects are supported by domestic institutions of higher learning which accounted for 74.55 per cent. Conclusion: The research support of the National Natural Science Foundation of China to the field of occupational diseases (H2402) has increased steadily, but the support of different research directions and supporting units is not balanced. It is suggested that departments concerned strengthen guidance and support for the applicants in less developed areas and weak research directions of the projects in the National Natural Science Foundation.

[Effects of glycosylated hemoglobin and disease course on islet ß-cell function in patients with type 2 diabetes].

OBJECTIVE: To compare islet ß-cell function in type 2 diabetic (T2DM) patients with different glycosylated hemoglobin (HbA1c) levels and diabetes durations. METHODS: We examined body parameters, biochemical profiles and islet autoantibodies in a total of 803 T2DM patients admitted in the Department of Endocrinology of the First Affiliated Hospital of Nanjing Medical University between December, 2014 and April, 2016. The patients were stratified by HbA1c level and disease course and underwent steamed bun test to evaluate islet ß-cell function and insulin resistance. RESULTS: Linear correlation analysis showed that in T2DM patients, HbA1c level was negatively correlated with HOMA2-IR, HOMA2-%ß, DI30 and DI180 (P=0.000), and disease course was negatively correlated with HOMA2-IR, HOMA2-% ß, and DI180 (P < 0.05). The patients with different HbA1c levels showed significantly different HOMA2-IR, HOMA2-%ß, DI30 and DI180 (P=0.000); HOMA2-%ß, DI30 and DI180 were significantly higher in patients with HbA1c levels < 7.8%, and HOMA2-% ß was significantly decreased in patients with HbA1c levels above 9.8%. The patients with different disease courses also had significant differences in HOMA2-IR, HOMA2-%ß, DI30, and DI180 (P=0.000), and as the disease course extended, DI30 and DI180 tended to decrease progressively. Multivariate linear regression analysis showed that HbA1c, diabetes duration, and body mass index (BMI) were all independent factors affecting islet ß- cell function in T2DM patients. CONCLUSIONS: The secretion function of islet ß cells decreases progressively with the increase of HbA1c level or disease course in T2DM patients, but the disease course does not appear to have an effect as strong as that of HbA1c level on islet ß cell function.

[The basic characteristics and medical status of pneumoconiosis patients under different investigation methods].

Objective: To analyze the characteristics of pneumoconiosis patients and the basic status of medical treatment. Methods: Research objects were chosen by stratified sampling method and typical survey method from existing pneumoconiosis patients in China. The survey was carried out from March 2017 to January 2018 in nine provinces including provinces from east, medium and western region in China. Source of pneumoconiosis cases were inpatient cases, outpatient or physical-examined cases and household-investigation cases. The survey mainly included demographic and sociological characteristics, economic status, occupational history and dust exposure history, disease status, work-related injury insurance and social security status and related indicators of pneumoconiosis treatment. Results: Investigated 1037 pneumoconiosis cases which included 186 (19.9%) household-investigation cases, 212 (20.4%) outpatient or physical-examined cases and 639 (61.7%) inpatient cases. Demographic and sociological characteristics, individual monthly income, economic source, occupational history and work-related injury insurance were statistically significant among different source of pneumoconiosis patients (P<0.05) . Among all of the household-investigation cases, there were 74 cases (40.2%) had no income, 117 cases (62.9%) used to work in private enterprises, 36 cases (19.4%) had work-related injuries insurance, 95 cases (51.1%) were at three phase of pneumoconiosis, 108 cases (59.0%) haven't had any drugs for pneumoconiosis. 65 cases (39.4%) haven't went to the clinic, 53 cases (28.5%) hadn't seek medical advice although they needed medical treatment very much. Among all of the outpatient or physical-examined cases, there were 95 cases (46.1%) had no income, 36 cases (17.0%) had work-related injuries Insurance, 139 cases (65.6%) went to the clinic for treatment of pneumoconiosis, 81 cases (38.2%) went to the clinic for more than ten times. Among all the inpatient cases, 310 cases' (49.3%) personal monthly income was above 2000 yuan, 352 cases (55.1%) had work-related injuries Insurance, 588 cases (92.2%) were taking drugs for treatment of pneumoconiosis, 153 canses (24.2%) had hospitalization for than ten times. Conclusion: Household-investigation cases have lower economic conditions, lower rates of Insurance coverage for work-related injuries, severer pneumoconiosis and higher clinical service utilization. Clinical or physical-examined cases have lower economic conditions, lower rates of Insurance coverage for work-related injuries and higher clinical service utilization. Hospitalized cases have better economic conditions, higher rates of insurance coverage for work-related injuries and higher hospitalization service utilization.

Role of MicroRNA 146a in Regulating Regulatory T Cell Function to Ameliorate Acute Cardiac Rejection in Mice.

BACKGROUND: MiR-146a plays a critical regulatory role in the homeostasis and function of regulatory T cells (Treg cells). The purpose of this study was to investigate the role of miR-146 in regulating Treg function to ameliorate acute cardiac rejection in mice. METHODS: We downregulated the miR-146a expression of Treg cells in recipients and constructed murine heterotopic heart transplantation models. Flow cytometry was used to analyze phenotypes of T cells. Graft rejection and cytokine changes during allograft rejection were detected. The changes of miR-146a target genes in Treg cells were detected by quantitative real-time polymerase chain reaction and western blotting. RESULTS: Our results indicated the miR-146a antagomir increased the Treg proportion in splenocytes and blood cells in the normal immune and inflammatory conditions, but impaired the ability of Treg cells to inhibit Th1 in the inflammatory condition. During the rejection process of murine heterotopic heart transplantation, miR-146a was mainly involved in regulating Treg cells through the STAT1/IFN-γ signaling pathway and the target gene was STAT1. Reducing the miR-146a of Treg and simultaneously collaboratively applying IFN-γ neutralizing antibodies can inhibit the proliferation of CD4+ T cell, alleviate the rejection process, and promote the survival of the donor's heart. CONCLUSION: It is possible to enhance the function of Treg cells in vivo of recipient by regulating a single miRNA, miR-146a, to achieve the purpose of inhibiting graft rejection. Collaboratively applying immunosuppressive drugs may effectively control the immune rejection and allow for the dosage and side effects of drugs to be reduced, a process that needs further study.

Biofilm Formation Plays a Role in the Formation of Multidrug-Resistant Escherichia coli Toward Nutrients in Microcosm Experiments.

In this study, microcosms were established to determine the effect of nitrogen (N) and phosphorus (P) on the multidrug resistance and biofilm-forming abilities of Escherichia coli. The expression of biofilm-formation-related genes was detected to establish correlations between genotype and phenotype. Different concentrations of N and P were added to make one control group and four treatment groups. The glass tube method was used to determine biofilm-forming capabilities. Real-time PCR was used to detect the mRNA abundance of six biofilm-formation-related genes in E. coli. No resistant strains were isolated from the control group; meanwhile, multidrug resistance rates were high in the treatment groups. Expression of the biofilm-associated genes luxS, flhD, fliA, motA, and fimH was detected in all treatment groups; however, there was no expression of mqsR. The expression of luxS, flhD, fliA, motA, and fimH significantly correlated with the concentration of N and P, as well as with the appearance and duration of multidrug resistance in different groups. Overall, the results of this study suggest that biofilm-forming ability plays a key role in the formation of multidrug resistance in E. coli after the addition of N and P to a microcosm.

[Clinic features of laryngeal carcinosarcoma and sarcomatoid carcinoma].

Objective: To investigate the clinic feature, pathology, therapy and prognosis of the sarcomatoid caricinoma or carcinosarcoma of the larynx. Methods: We reviewed the clinical records of 7 patients with laryngeal carcinosarcoma /sarcomatoid caricinoma who were treated at our hospital between June 1996 and August 2016. All patients were men (mean age, 65.9 years; range, 52 to 94 years). Among 7 patients, 6 had a history of smoking; 2 underwent radiotherapy; and 5 patients who didn't undergo radiotherapy complained of hoarseness. The glottis was the most frequent site of involvement. Most tumors exhibited a polypold or pedunculated gross morphology. Among the 5 patients who didn't undergo a radiotherapy, 2 were in stage Ⅰ, 2 in stageⅡ, and 1 in stage Ⅲ. The other 2 cases underwent surgeries and radiotherapy were staged. Results: All 7 patients received surgeries, without lymph node metastasis. All the tumors were pathologically carcinosarcoma/sarcomatoid carcinoma. With immunohistochemistry examination, Vimetin was positive in all tumors, SMA positive in 3 tumors, S-100 positive in 1 tumors, but CD-68, HMB-45 or Myglobin was negative in all tumors. With follows-up from 3 months to 20 years, of 7 patients, 4 survived without recurrent, 1 dead, and 2 lost connection. Conclusions: Both of the carcinosarcoma and the sarcomatoid carcinoma of larynx contain pathologically carcinoma and sarcoma. Surgery is the best choice for laryngeal sarcomatoid carcinoma, and these patients without a undergoing radiotherapy before surgery or these with little sarcoma in tumors show better prognosis.

[The significance of 8-hydroxy-deoxyguanosine acid in the diagnosis of nonalcoholic steatohepatitis].

Objective: To evaluate the significance of serum 8-hydroxy-deoxyguanosine acid(8-OHdG) in the diagnosis of nonalcoholic steatohepatitis (NASH). Methods: Patients or healthy subjects were enrolled at the Second Hospital of Tianjin Medical University and the Second People's Hospital of Tianjin from May 2013 to December 2015. A total of 41 patients with nonalcoholic fatty liver disease were enrolled in the study, including 20 nonalcoholic simple fatty liver (NAFL) patients and 21 NASH patients whose diagnosis were proven by liver biopsy. The other 32 healthy subjects were studied as controls. Serum 8-OHdG, ALT, AST and GGT were tested. Nonalcoholic fatty liver disease activity score (NAS) and expression of 8-OHdG in liver was investigated between NAFL patients and NASH patients. The correlations between serum 8-OHdG and serum ALT, AST, GGT, and 8-OHdG in liver tissue in NASH group were investigated. In addition, the receiver operating characteristic (ROC) curve analyses for ALT and 8-OHdG levels were performed in NAFL patients and NASH patients, and the cut-off value was determined. Results: Serum 8-OHdG values in healthy controls, NAFL and NASH patients were (0.19±0.16) µg/L, (0.22±0.16) µg/L, (0.42±0.21) µg/L respectively. The serum 8-OHdG and serum ALT, GGT and 8-OHdG in liver tissue were all positively correlated in NASH group with respective correlation coefficient r values as 0.454 7, 0.382 9, and 0.497 6. AUC of 8-OHdG was 0.901 with cut-off value 0.39 µg/L. Its sensitivity was 88.3% and specificity was 81.5%, which were higher than those of ALT. Conclusion: The value of serum 8-OHdG would be used as a marker for the diagnosis of NASH.

Experimentally simulating quantum walks with self-collimated light.

In self-collimated photonic crystal, periodically arranged air holes of sub-wavelength scale provide flattened equi-frequency curves perpendicular to the ΓM direction, which allow light or photons propagating in a quasi-uniform medium without diffraction. Here we for the first time experimentally simulate four-step single-photon discrete time quantum walks with classical light in such a photonic crystal chip fabricated on silicon-on-insulator. Similarities between theoretical expectations and experimental results are higher than 0.98. The functional area is compact and can be extended to construct more complicated linear quantum circuits.

Expression of TRPM8 in diabetic rats and its relationship with visceral pain stimulation.

Transient receptor potential cation channel, subfamily M, member 8 (TRPM8) is a nonselective cation channel and a candidate for cold sensation signaling, but the relationship between TRPM8 and diabetes remains unclear. In the present study, we determined the expression levels of TRPM8 messenger RNA (mRNA) and the levels of the TRPM8 protein in the bladder tissue of diabetic rats. We also investigated the correlation between TRPM8 expression and the visceral pain stimulation-related factor, calcitonin gene-related peptide (CGRP) in diabetic rats. The rats were sacrificed 3, 5, 7, and 15 days after streptozotocin injection, and blood was collected from their tail veins to determine the blood glucose levels. Bladder tissue was removed to assess the expression of TRPM8 mRNA by reverse transcription-polymerase chain reaction, and the expression of the TRPM8 protein by western blotting. After administering electrical stimulation (5 V/1 Hz), the expression levels of TRPM8 and CGRP proteins were determined. Our results revealed that the blood glucose level, and TRPM8 mRNA and TRPM8 protein expression levels increased significantly in the diabetic rats. Spinal tissue protein expression levels of both TRPM8 and CGRP also increased significantly following electrical stimulation. This possibly indicates that TRPM8 is closely associated with visceral pain stimulation, and could be an independent prognostic biomarker for diabetes.

Improved Cuff Technique for Establishing a Mouse-Rat Heterotopic Cardiac Xenotransplantation Model.

BACKGROUND: The small animal model of cardiac transplantation is the most common model in organ transplantation studies. The cervical heterotopic transplantation is widely performed because this allows for direct observation of the graft heartbeat and contributes to early prediction of graft rejection. OBJECTIVE: A mouse-rat cervical heterotopic cardiac xenotransplantation model was modified with respect to the anesthesia method, cardiac graft harvesting method, and perioperative treatment. These improvements ensure the stability and reliability of xenotransplantation models for in vivo studies of immune-mediated graft rejection. METHODS: After establishing isoflurane inhalation anesthesia, the donors' hearts were harvested. The experimental method involved separate ligation of the left and right superior venae cavae; the other blood vessels were ligated in a cluster. Both the donor and recipient animals were placed on a heating pad intraoperatively to maintain a body temperature of 37-40 °C. The model establishment was divided into 3 stages: practice, stabilization, and stereotyping. The surgical success rate and operation time were recorded. Specimens were harvested at different time points for histopathological examination. RESULTS: The anesthetic effect of isoflurane was well maintained, and no animals died of adverse anesthetic events. Body temperature was maintained at 37-40 °C which effectively shortened the time to restoration. The modification of the cardiac graft harvesting method is conducive to rebeating of the donor heart. The success rates in the stabilization and stereotyping stages were significantly higher than that in the practice stage (P < .05). The operation time in the stabilization and stereotyping stages were significantly shorter than those in the practice stage (P < .05). Histopathological examination revealed thrombosis formation, interstitial hemorrhage, and inflammatory cell infiltration in the donor hearts. CONCLUSION: Our findings suggest that the mouse-rat cervical heterotopic cardiac xenotransplantation model is the ideal animal model for studying xenograft rejection.