A decision tree model to distinguish between benign and malignant pulmonary nodules on CT scans.

OBJECTIVE: Chest computed tomography (CT) is increasingly being used to screen for lung cancer. Machine learning models could facilitate the distinction between benign and malignant pulmonary nodules. This study aimed to develop and validate a simple clinical prediction model to distinguish between benign and malignant lung nodules. PATIENTS AND METHODS: Patients who underwent a video thoracic-assisted lobectomy between January 2013 and December 2020 at a Chinese hospital were enrolled in the study. The clinical characteristics of the patients were extracted from their medical records. Univariate and multivariate analyses were used to identify the risk factors for malignancy. A decision tree model with 10-fold cross-validation was constructed to predict the malignancy of the nodules. The sensitivity, specificity, and area under the curve (AUC) of a receiver operatic characteristics curve were used to evaluate the model's prediction accuracy in relation to the pathological gold standard. RESULTS: Out of the 1,199 patients with pulmonary nodules enrolled in the study, 890 were pathologically confirmed to have malignant lesions. The multivariate analysis identified satellite lesions as an independent predictor for benign pulmonary nodules. Conversely, the lobulated sign, burr sign, density, vascular convergence sign, and pleural indentation sign were identified as independent predictors for malignant pulmonary nodules. The decision tree analysis identified the density of the lesion, the burr sign, the vascular convergence sign, and the drinking history as predictors of malignancy. The area under the curve of the decision tree model was 0.746 (95% CI 0.705-0.778), while the sensitivity and specificity were 0.762 and 0.799, respectively. CONCLUSIONS: The decision tree model accurately characterized the pulmonary nodule and could be used to guide clinical decision-making.

[Influencing factors of poor treatment adherence in uncontrolled asthma patients in China].

Objective: To evaluate the influencing factors of poor treatment adherence in patients with uncontrolled asthma in China. Methods: From April 2017 to April 2018, all asthma patients with uncontrolled asthma and poor compliance in 32 third-class hospitals in 28 provinces and cities of China mainland included in the "National Mobile Asthma Assessment and Management Project" were selected as the subjects. A total of 923 patients were enrolled in the study including 388 males and 535 females. By analyzing the baseline data of the patients at the initial visit when enrolled, the influencing factors of poor adherence of adult asthma was analyzed by inter-group comparison and χ2 test. Results: Poor compliance in asthma was related to the following factors: age from 59 to 68 years old, course of disease more than 20 years, low education level, non-local follow-up, having obstructive ventilation dysfunction and low awareness of the disease[P values were 0.026(t=1.20), 0.004(t=3.97), 0.001(t=4.92), 0.003(t=3.98), 0.032(t=1.22) and 0.001(t=4.99), respectively]. Totally, 243 patients (26.33%) answered all the questions about asthma correctly. Their medication adherence rating scale (MARS-A) scores were significantly higher than those who answered incompletely correctly (36.23±5.85 vs. 31.77±5.74, P=0.001). Conclusions: The adherence of adult asthma patients was affected by individual and external environment factors. Clinicians should choose individualized methods based on the characteristics of patients. Patient education should be strengthened to improve patients' awareness of the disease at the same time.

[Incidence and cause of abnormal cholesterol in children aged 2-18 years in a single center].

OBJECTIVE: To investigate the abnormality and distribution of plasma cholesterol levels in single-center hospitalized children. METHODS: The blood lipid levels of children aged 2-18 years who had blood lipid test results in Peking University First Hospital from June 2016 to June 2019 were etrospectively analyzed. Cholesterol oxidase method was used for total cholesterol, and high-density lipoprotein cholesterol and low-density lipoprotein cholesterol were detected by clearance method. The counting data were compared with chi-square test. RESULTS: The survey had involved 11 829 children (7 087 were boys and 4 742 were girls). 1 822 (15.4%) children were with elevated total cholesterol, 1 371 (11.6%) children with elevated low-density lipoprotein cholesterol, and 2 798 (23.7%) children with high-density lipoprotein cholesterol reduction. The total number of the children with abnormal cholesterol levels was 4 427 (37.4%). Among the 7 835 children who visited hospital due to the disease not commonly inducing dyslipidemia, 731 (9.3%) had elevated TC, 561 (7.2%) had elevated LDL-C, 1 886 (24.1%) had decreased HDL-C, and 2 576 (32.9%) had abnormal cholesterol levels. Among the children with different diseases, the difference in the incidence of abnormal cholesterol was statistically significant. The top three main groups of the children with increased total cholesterol and low-density lipoprotein cholesterol were "dyslipidemia", "urinary tract disease", and "nutritional disease"; The top three main groups of the children with reduced high-density lipoprotein cholesterol were "respiratory diseases", "dyslipidemia", "hematological diseases and malignant tumors". Among the 1 257 blood li-pid test results sent by other departments, 300 cases had abnormal cholesterol levels (23.8%). Among them, there were 70 children with hypercholesterolemia (5.6%), 44 children with increased low-density lipoprotein cholesterol (3.5%), and 224 children with reduced high-density lipoprotein cholesterol (17.8%). There were 365 (4.6%) children with low-density lipoprotein cholesterol ≥140 mg/dL (3.6 mmol/L) who needed to further exclude familiar hypercholesterolemia among the children who visited hospitals due to the disease not commonly inducing dyslipidemia. CONCLUSION: Children in hospitals have a high incidence of cholesterol abnormalities. Doctors need to pay more attention to the cholesterol diagnosis and management regardless of the discipline, which not only helps to control secondary hypercholesterolemia, but also provides the possibility of detecting familial hypercholesterolemia in time.

Advances in research on the relationship between the gut microbiome and cancer.

OBJECTIVE: The objective of this review is to provide currently available information on the relationship between the gut microbiome and cancer. MATERIALS AND METHODS: In this mini-review, we explored the PubMed, EMBASE, and Google Scholar electronic databases, with regards to the searching terms "gut microbiome, cancer, intestinal flora, immunotherapy, immune checkpoint inhibitor". By reviewing and analyzing the literature, we analyzed how the bacterial microbiome influences the immune system and cancer, as well as how changes in symbiotic flora may be applied to improve the efficacy of cancer immunotherapy. RESULTS: The microbiota is related to the development of tumors and may promote canceration. In recent years, a number of studies have confirmed the influence of intestinal flora on immune checkpoint inhibitors in cancer patients, and studies have also shown the link between the intestinal microbiome and treatment-related immune toxicity. Antibiotics, proton pump inhibitors, and hormones affect the composition of the gut microbiota. CONCLUSIONS: Intestinal flora is closely related to cancer. Intestinal flora has a certain impact on cancer occurrence, cancer treatment, cancer immunotherapy efficacy, and side effects.

[Investigation on status of lung cancer cough management and concept and practice of lung cancer cough management in Chinese medical providers].

Objective: Investigate the current situation of lung cancer cough diagnosis and treatment and the awareness of related issues among Chinese medical providers. Methods: Doctors, nurses, pharmacists from the oncology department, respiratory department, or general department were investigated using an electronic questionnaire from January 29 to March 29, 2021. There were 25 questions about lung cancer in the questionnaire. The questionnaire was including the most common accompanying symptoms in patients with lung cancer, the incidence of lung cancer-related cough, the assessment of the proportion of central antitussive drugs, and the commonly used central antitussives. Results: Questionnaires from 2 424 medical providers were collected from 402 hospitals in 21 provincial administrative units. Cough was the most common symptom in lung cancer. Most physicians believed that the incidence of lung cancer-related cough was 51%~75%, while the proportion of patients satisfied with the treatment was only 11%~20%. The evaluation of lung cancer-related cough was seriously insufficient. The leading cause of lung cancer-related cough was tumors. And the main problem was the inadequate antitussive effect of drugs in lung cancer-related cough management. The proportion of central antitussive medication usage in the secondary and tertiary hospitals was 93.9% and 92.0%, significantly higher than 75.0% in Primary hospitals (χ²=8.390, P=0.015). The proportion of the physicians who underhanded that codeine is at risk of addiction was 76.6% and 72.0% in the secondary and tertiary hospitals, which were significantly higher than 53.9% in Primary hospitals (χ²=9.240, P=0.010). In different occupations, the proportions of doctors and pharmacists who knew the risk of addicting to codeine were 73.0% and 82.6%, which were significantly higher than the 66.4% of nurses (χ²=21.200, P<0.001). The Chinese medical providers were lack of training about the basic knowledge of using central antitussive medication. Conclusions: The proportion of patients who were satisfied with the effect of cough treatment is low. The medical staff did not have enough awareness of this. There was an urgent need to develop a consensus and standardize lung cancer cough diagnosis and treatment in China.

[A prospective multi-center clinical investigation of HIV-negative pulmonary cryptococcosis in China].

Objective: To investigate the current status of the diagnosis and treatment of pulmonary cryptococcosis in respiratory medicine and improve the understanding of the clinical characteristics of HIV-negative pulmonary cryptococcosis in China. Methods: A prospective multi-center open cohort study was designed to screen for pulmonary cryptococcosis in the general wards and intensive care units of the Department of Respiratory Diseases in 22 hospitals. The HIV-negative patients with positive cryptococcal etiological diagnosis based on smear culture, antigen detection and histopathology were enrolled in the study. The clinical data of enrolled patients were collected and analyzed. Results: A total of 457 cases of pulmonary cryptococcosis were enrolled, among which 3.28% (15/457) were disseminated infections. The case fatality rate was 0.88% (4/457). The majority of the cases were diagnosed by histopathological examinations (74.40%, 340/457) and cryptococcus antigen detection (37.64%, 172/457). Patients with pulmonary cryptococcosis accounted for 2.04‰ (457/223 748) of the total hospitalized patients in the Department of Respiratory Diseases during the same period, and the ratio was the highest in south and east China. Meanwhile, 70.24% (321/457) of the patients had no underlying diseases, while 87.75% (401/457) were found to have immunocompetent status. Cough and expectoration were the most common clinical symptoms in patients with pulmonary cryptococcosis. However, 25.16% (115/457) of the patients had no clinical symptom or physical signs. In terms of imaging features on pulmonary CT, multiple pulmonary lesions were more common than isolated lesions, and there were more subpleural lesions than perihilar or medial lesions. Morphologically, most of the lesions were middle-sized nodules (1-5 cm) or small-sized nodules (3 mm to 1 cm). The sensitivity of serum cryptococcus antigen test was 71.99% (203/282). Moreover, antigen-positive patients differed from antigen-negative patients in terms of basic immune status, clinical symptoms, imaging features and infection types. Meanwhile, immunocompromised patients differed from immunocompetent patients in terms of clinical symptoms, physical signs, infection-related inflammation indicator levels, imaging features, serum cryptococcus antigen positive rate and prognosis. Conclusions: The majority of cases of HIV-negative pulmonary cryptococcosis in China had no underlying disease or immunocompromised status, and the overrall prognosis was favorable. However, early diagnosis of HIV-negative pulmonary cryptococcosis remains challenging due to the complicated manifestations of the disease.

[Efficacy and safety of Kangbingdu granules in the treatment of influenza: a randomized, double-blind, double-dummy, positive-drug parallel control multicenter clinical trial].

Objective: To observe the efficacy and safety of Kangbingdu granules (KBD) in the treatment of influenza. Methods: A multicenter, randomized, double-blind, double-dummy, and positive-drug parallel control trial was conducted in 27 Grade ⅢA hospitals in China and the subjects were randomly assigned to the KBD test group or the oseltamivir phosphate capsule control group at a ratio of 1∶1. 200 subjects were planned to be enrolled in each group. The experimental group was given KBD (18g each time, 3 times a day) and oseltamivir phosphate simulator orally, while the control group was given oseltamivir phosphate capsule (75 mg each time, twice a day) and KBD simulator orally for 5 days. The primary efficacy indicators included the remission time of major clinical symptoms and the time of complete defervescence. The secondary efficacy indicators included dosage of acetaminophen, the change of traditional Chinese medicine (TCM) syndrome score and the remission time of other important clinical symptoms. The efficacy of KBD in the test group and Oseltamivir phosphate control group were compared. Adverse events or adverse reactions were observed at the same time to evaluate the safety of KBD Granules. Results: A total of 393 subjects from 27 Grade ⅢA hospitals in China were enrolled. The experimental group included 195 subjects and 191 subjects (97.95%) completed the trial, While the control group included 198 subjects and 195 subjects (98.48%) completed the trial. There was no significant difference in the shedding rate and rejection rate between the two groups (P>0.05). In the Full Analysis Set (FAS), the mean age of the experimental group was (34.9±14.4) years old, with 83 males (42.78%). The mean age of the control group was (33.3±13.5) years old, with 78 males (39.59%). There were no statistically significant differences between the two groups in demographic data, physical examination, viral pathogen detection, total score of TCM syndromes and scores of each symptom at baseline (P>0.05). In the FAS, the remission time M (Q1, Q3) of major clinical symptoms was 3.0 (3.0, 4.0) days in the experimental group and 3.0 (3.0, 4.0) days in the control group, and the difference was not statistically significant (P>0.05). The time M (Q1, Q3) of complete defervescence was 34.0 (20.3, 49.0) hours in the experimental group and 36.5 (19.6, 48.8) hours in the control group, and the difference was not statistically significant (P>0.05). KBD granules had the same effect as Oseltamivir phosphate capsule (P>0.05) in terms of acetaminophen dosage, TCM syndrome effect and disappearance rate of most important clinical symptoms. Meanwhile, the disappearance rate of dizziness and chest distress on day 3 in the KBD granules group was better than that of oseltamivir phosphate capsule (P<0.05). Conclusion: KBD granules have the same efficacy as Oseltamivir Phosphate capsule in the treatment of influenza and the drug safety is good.

[Neuronal intranuclear inclusion disease: a clinicopathological study of two cases].

Objective: To investigate the clinical manifestations, imaging features, histopathologic, immunohistochemical (IHC) and ultrastructure features of neuronal intranuclear inclusion disease (NIID). Methods: HE, IHC staining and EM were performed in cases of NIID diagnosed at the Department of Pathology, Jinling Hospital from 2018 to 2019. Results: Two cases were identified, including one male and one female; both patients were 76 years old. They were hospitalized because of nervous system dysfunction. MRI showed abnormal high signal intensity in corticomedullary junction of bilateral frontal lobes (male patient) and bilateral cerebral hemispheres (female patient). Light eosinophilic transparent inclusion bodies were seen in the nuclei of neurons in both rectal mucosa and cutaneous sweat glands, and these were positive for p62 by IHC. By scanning EM, the inclusion bodies in the sweat gland cells nuclei were round membranous structures consisting of 8-18 nm microfilaments. Conclusions: NIID is a rare, multi-system and slowly progressive neurodegenerative disease. Its clinical manifestations are highly diverse and easily misdiagnosed or missed. Neuroimaging can make a preliminary diagnosis. In the past, NIID can only be diagnosed through autopsy, and this study demonstrates that NIID can be confirmed through skin or rectal mucosal biopsy.

Butorphanol attenuates inflammation via targeting NF-κB in septic rats with brain injury.

OBJECTIVE: To observe the therapeutic effect of butorphanol on brain tissue injury in rats with sepsis through the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway. MATERIALS AND METHODS: Sprague-Dawley rats were divided into control group (n=20), sepsis model group [cecal ligation and perforation (CLP) group, n=20], and butorphanol treatment group (n=20). After successful modeling, the blood and brain tissues were collected from rats at 24 h. The content of serum brain injury indexes was detected. Hematoxylin-eosin (HE) staining assay and enzyme-linked immunosorbent assay (ELISA) were separately carried out to observe the pathological changes and measure the levels of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and myeloperoxidase (MPO) activity. The neurological function was scored in rats. Glial fibrillary acidic protein (GFAP), S100, and NF-κB signaling pathway genes and proteins in brain tissues were detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) and Western blotting. RESULTS: CLP group showed remarkably increased levels of serum glucosuria (GLU), creatinine (CR), and Na+ but an evidently reduced level of K+ in comparison with the control group (p<0.05), while the treatment group displayed contrary trends. Histopathological observations showed that the rats in the CLP group suffered a brain injury, while those in the treatment group had mild pathological changes. The MPO in the CLP group was significantly increased compared with that in the control group (p<0.05). The levels of TNF-α and IL-6 were overtly higher in the CLP group than those in the control group, and these indexes in the treatment group were close to those in the control group. The messenger ribonucleic acid (mRNA) expression levels of S100, GFAP, Toll-like receptor 2 (TLR2), and NF-κB in CLP group were evidently higher than those in the control group and treatment group (p<0.05). The results of Western blotting revealed that the protein expression of NF-κB was significantly higher in CLP group than that in the control group, and it declined in the treatment group, which was close to that in the control group. CONCLUSIONS: Butorphanol can reduce the content of inflammatory factors TNF-α, IL-1, and IL-6 through the NF-κB signaling pathway, thereby relieving the brain injury caused by sepsis.

[Clinical analysis of Langerhans cell histiocytosis originating in the base of nasal skull].

Objective:To summarize the clinical features, diagnosis and treatment of Langerhans histiocytosis（LCH） which first appeared in the nasal skull base. Method:Ten cases of LCH with nasal and skull base symptoms were analyzed retrospectively. The clinical characteristics of LCH with nasal and skull base symptoms were summarized. The correlation of other systems involved in LCH was analyzed. Result:Among the 10 patients, the youngest was 1 year and 5 months, and the oldest was 8 years, the average age was 3 years. The main imaging manifestations were osteolytic changes and soft tissue invasion. Seven patients were monofocal and three patients were multifocal. For localized lesions, radical resection and follow-up chemotherapy were performed, and conservative treatment was performed for patients with multiple system involvement and obvious systemic symptoms. Eight patients survived, 2 died. Conclusion:LCH occurs frequently in children and has certain clinical characteristics. Single system and single lesion surgery have a better therapeutic effect, and can achieve a greater survival rate with follow-up chemotherapy.

[Adult peripheral primary neuroectodermal tumor: a case report and literature review].

Objective: To analyze the clinical features, diagnosis and differential diagnosis of peripheral primary neuroectodermal tumor(pPNET). Methods: The clinical data and diagnosis of a patient with rapid progressive pPNET in Shandong Provincial Hospital affiliated to Shandong University in January 2016 was reported and the related literatures were reviewed.The literature reviews were carried out respectively in CNKI, Wanfang and PubMed by July 2016 with "primitive neurotodermal tumour" and "PNET" being the search term from March 1994 to July 2016, including 13 articles. Results: A 41 year-old male patient was admitted to the hospital because of shortness of breath and occasional chest tightness, accompanied by general asthenia of about 15 d. Positron emission and transmission-CT of total trunk showed a mass in the right femoral osteoperiosteal mass, and multiple nodules in the left lung, lumps and nodules in the right lung, and right pleural thickening and effusion. Thoracoscopy was performed and pathology study confirmed the diagnosis of peripheral primary neuroectodermal tumor. The patient was given 2 courses of chemotherapy, but had rapid progressive worsening and died 1 month after PNET diagnosis. A total of 13 literatures of PNET were retrieved, all of which were case reports, and a total of 15 cases were reported.There were 9 male and 7 female patients, with a median age of 26 years.The symptoms had no specificity and most of them were solid masses in chest images, with or without pleural effusion. Eight cases were diagnosed by imaging guided percutaneous biopsy, 7 by operation and pathology, and 1 by medical thoracoscopy. Four cases underwent chemotherapy, and the survival time was less than 6 months. Twelve cases got surgical resection: 2 with surgical treatment, 7 with postoperative chemotherapy, 2 with postoperative radiotherapy and chemotherapy, 1 with postoperative chemotherapy and autologous bone marrow transplantation treatment, among which 11 patients completed follow-up. During follow-up, 3 cases died, and the survival time was 10 months, 3 years and 7 years, respectively. Conclusions: PNET is rare.Due to the lack of specificity of clinical manifestations, clinical diagnosis depends on biopsy. Early diagnosis and surgical resection are especially important for prognosis and quality of life.

[The role of HuR in mediating snail expression in human small airway epithelium induced by cigarette smoke extract].

Objective: To investigate the abundance of human antigen R (HuR) in small airway epithelial cells stimulated by cigarette smoke extract (CSE) as well as the role of HuR in mediating snail which is recognized as a key transcription factor in regulating epithelial-mesenchymal transition (EMT). Methods: Human small airway epithelial cells (HSAEpiC) cultured in vitro were exposed to cigarette smoke extract (CSE) to model COPD status. Real-time PCR and western blotting analysis were used in detecting HuR protein and mRNA expression in cells with CSE which were divided into 5 groups: a control group, a 1%-24 h group, a 3%-24 h group, a 1%-48 h group and a 3%-48 h group. Small interfering RNA (siRNA) transfection was used to decrease HuR abundance. HuR expression at both mRNA and protein levels was detected using Real-time PCR and Western blotting analysis respectively, and the experiment was divided into 3 groups: a control group, a transfection control group and a transfection group. Snail, E-cadherin and vimentin levels were determined using Western blotting test in cells with both CSE exposure and HuR siRNA transfection which were divided into three groups: control group, CSE group and CSE + transfection group. Results: After CSE stimulation, HuR expression was increased at both mRNA and protein levels [mRNA 1%-24 h group (1.12±0.04), 3%-24 h group (1.41±0.06), 1%-48 h group (1.26±0.05), 3%-48 h group (1.49±0.06), protein 3%-24 h group (1.35±0.08), 1%-48 h group (1.17±0.06), 3%-48h group (1.42±0.06) all P<0.05]. Compared with the control siRNA, after HuR siRNA transfection, HuR mRNA and protein levels were significantly reduced [mRNA level (0.33±0.06) vs (1.02±0.10), protein level (0.46±0.07) vs (0.97±0.06), all P<0.01]. Control siRNA transfection had no effect on HuR expression [mRNA level (1.02±0.10), protein level (0.97±0.06), all P>0.05]. After 48 h stimulation with 3% CSE, the expression of HuR (1.47±0.11), snail (1.46±0.05) and vimentin (1.56±0.05) increased and the expression of E-cadherin (0.49±0.05) decreased. After transfection and CSE stimulation, the expression of HuR (0.84±0.06), snail (1.22±0.06) and vimentin (1.11±0.09) decreased and the expression of E-cadherin (0.73±0.06) increased. (All P>0.05). Conclusions: CSE promoted the expression of HuR in human small airway epithelial cells. HuR participated in the regulation process of EMT key transcription factor snail and might regulate EMT process by this action.

Two Distinct Genotypes of Porcine Epidemic Diarrhoea Virus in Vaccinated Pig Flocks in Shandong Province of China, 2012-2015.

As one of the top pork producers in China, Shandong Province suffered frequent outbreaks of porcine epidemic diarrhoea virus (PEDV) on pig farms from January 2012 to July 2015, resulting in significant economic losses. To better understand the prevalence situation, we conducted molecular epidemiological analyses of 38 PEDV strains isolated from 13 cities in Shandong Province. The detection rate of PEDV was 71.2% (146/205) by reverse transcription polymerase chain reaction (RT-PCR). The S genes of the 38 isolated samples were 4146 to 4161 nt in length and shared high levels of sequence identity (93.3-99.6% nt, 92.1-99.4% aa) with those of the 41 reference strains. Among the 38 strains, 31 strains that occupied 12 cities were classed into G3 genotype, while the other seven that only existed in four cities were classed into G2 genotype. In addition, the strains CH-SDLY-2-2014 and CH-SDLY-3-2014 isolated from Linyi were classed into the Gd subgenotype. Notably, there were multiple insertions or deletions in the S genes and several mutations in the neutralizing epitopes of the PEDV S protein. Overall, the results revealed that G2 and G3 are the predominant PEDV genotypes circulating in Shandong Province during 2012-2015, and Gd subgenotype in G3 group had already spread towards northern China in 2014.

[Human antigen R regulates TGF-ß1 expression in airway smooth muscle cells under the stimulation of platelet-derived growth factor].

Objective: To study the role of human antigen R (HuR) regulated transforming growth factor ß1 (TGF-ß1) expression in airway smooth muscle cells under the stimulation of platelet-derived growth factor (PDGF). Methods: Airway smooth muscle (ASM) cells were cultured at 37 ℃ and 5% CO2 in dulbecco's modified eagle medium (DMEM) cell medium. Cells at passages between 4 and 11 were divided into different groups according to the different compounds added. For control group, no compounds were administrated. For PDGF group, cells were stimulated with PDGF (20 µg/L) and cultured for an additional time. Cells were harvested and real-time PCR was used to measure mRNA level and Western blotting to detect protein level of HuR and TGF-ß1 in ASM cells for different groups. Cells were divided into HuR siRNA group and control group. RNA-interference was used to determine whether lowering HuR expression could decrease PDGF-induced TGF-ß1 expression in HuR siRNA group and control group after the stimulation of PDGF for indicated times. Western blotting analysis was used to test the expression of TGF-ß1 after interrupting HuR expression. The concentration of TGF-ß1 in the cultured serum of HuR siRNA group and control group for 0, 6, 12 h under the stimulation of PDGF was measured by enzyme-link immunosorbent assay (ELISA). Cells were divided into control group, control+ PDGF 6 h group, HuR siRNA group and HuR siRNA+ PDGF 6 h group, then the half-life of TGF-ß1 mRNA in different groups was determined by treating ASM cells with the transcriptional inhibitor actinomycin D (10 mg/L) for 0, 4, 8 and 12 h. Results: PDGF treatment for 0, 6, 12 and 24 h significantly promoted HuR mRNA and protein expression and the relative levels were 1.00±0.00, 1.35±0.14, 1.73±0.17, 2.07±0.10; 0.51±0.10, 0.67±0.05, 0.83±0.07, 0.95±0.02 (all P<0.05). Similar alterations could also be demonstrated at TGF-ß1 mRNA and protein. The relative expression was 1.00±0.00, 1.27±0.06, 1.60±0.10, 1.87±0.10; 0.72±0.09, 0.87±0.07, 1.13±0.12, 1.33±0.05 (6 h versus 12 h and 12 h versus 24 h, P<0.05). HuR expression decreased 21.9% in HuR siRNA group compared with control group under the stimulation of PDGF for 12 h. HuR silencing also decreased PDGF-induced TGF-ß1 over-expression in ASM cells. In the control group, the relative protein levels of PDGF treatment for 0, 6 and 12 h were 0.70±0.05, 0.89±0.06, 1.06±0.05 and the protein levels in HuR siRNA group were 0.67±0.09, 0.77±0.03, 0.89±0.05 (all P<0.05). The concentration of TGF-ß1 in the cultured serum was measured by ELISA and the outcomes were (773.33±16.32, 877.97±16.03, 3 060.34±82.53) ng/L in control group for 0, 6 and 12 h. Under the same condition, the outcomes of HuR siRNA group were (277.33±9.93, 407.77±7.14, 828.05±11.67) ng/L (both P<0.05). Actinomycin D disturbed the process of transcription and the half-life of TGF-ß1 mRNA in HuR siRNA group showed no significant change compared with the HuR siRNA+ PDGF 6 h group (P>0.05). However, compared with control group and control + PDGF 6 h group, the half-life of TGF-ß1 mRNA in HuR siRNA group showed significant change (P<0.05). Conclusions: PDGF can elevate TGF-ß1 expression in ASM cells. HuR regulates TGF-ß1 expression by promoting its mRNA stability.

[Rapidly progressive pulmonary malignant perivascular epithelioid cell tumor: a case report and literature review].

Objective: To analyze the pathogenesis, clinical features, diagnosis and differential diagnosis of primary perivascular epithelioid cell tumor(PEComa). Methods: The clinical features, auxiliary examinations and diagnosis of a case with rapidly progressive pulmonary malignant PEComa were reported and the related literatures were reviewed.The literature review was carried out respectively in Wanfang Data, CNKI and PubMed from Jan. 1975 to Jul. 2015 with "pulmonary malignant perivascular epithelioid cell tumor" and "PEComa" being the search terms. Results: A 50 year-old female patient was admitted to the hospital on September 4, 2014 because of cough and dyspnea for 60 days, hemoptysis for 40 days and fever for 7 days.Chest CT scan showed diffuse small nodules with infiltrative border and multiple pure and mixed ground-glass opacity. Transbronchial lung biopsy (TBLB) was performed and the pathological study confirmed the diagnosis of primary pulmonary malignant PEComa. The patient declined further specific therapy, but followed by rapidly progressive respiratory failure, and died two weeks after the diagnosis. A total of 8 literatures were retrieved from Wanfang Data, CNKI and PubMed and all of them were case reports.There were 3 male and 5 female patients, aging from 50 to 79 years.Radiographically, the previously reported cases presented as round and well-circumscribed masses with or without multiple nodules in both lungs. The symptoms had no specificity. Conclusions: Pulmonary malignant PEComa is a rare disease.It is easily misdiagnosed because of non-specific clinical and imaging manifestations.The final diagnosis depends on pathological biopsy.TBLB is an effective diagnostic method.

Downregulation of SWI5 and CTC1 genes: hepatitis B virus DNA polymerase transactivated protein 1-mediated inhibition of DNA repair.

Hepatitis B virus (HBV) DNA polymerase transactivated protein 1 (HBVDNAPTP1) is a novel protein upregulated by HBV DNA polymerase, which has been screened by suppression subtractive hybridization technique (SSH) (GenBank Acc. No. AY450389). A vector pcDNA3.1 (-)/myc-His A-HBVDNAPTP1 was constructed and used to transfect acute monocytic leukemia cell line THP-1. HBVDNAPTP1 expression was detected by Western blot analysis in the cells. A cDNA library of genes downregulated by HBVDNAPTP1 in THP-1 cells was made in pGEM-T Easy using SSH. The cDNAs were sequenced and analyzed with BLAST search against the sequences in GenBank. Some sequences, such as DNA repair protein SWI5 homolog (SWI5) and CTS telomere maintenance complex component 1 (CTC1), might be involved in DNA repair. Protein expression of SWI5 and CTC1 was identified by Western blot in THP-1 cells. HBVDNAPTP1 could downregulate the expression of SWI5 and CTC1 at translation level.

Accurate LC peak boundary detection for ¹6O/¹8O labeled LC-MS data.

In liquid chromatography-mass spectrometry (LC-MS), parts of LC peaks are often corrupted by their co-eluting peptides, which results in increased quantification variance. In this paper, we propose to apply accurate LC peak boundary detection to remove the corrupted part of LC peaks. Accurate LC peak boundary detection is achieved by checking the consistency of intensity patterns within peptide elution time ranges. In addition, we remove peptides with erroneous mass assignment through model fitness check, which compares observed intensity patterns to theoretically constructed ones. The proposed algorithm can significantly improve the accuracy and precision of peptide ratio measurements.

Effectiveness of transient immunosuppression using cyclosporine for xenomyoblast transplantation for cardiac repair.

We studied the survival of human myoblast for cellular myocardial reconstruction in a porcine model of chronic myocardial ischemia with immune tolerance using transient immunosuppression. A porcine model of chronic cardiac ischemia was created in 10 pigs (DMEM medium-injected n = 4; myoblast transplanted n = 6) by clamping ameroid ring around left circumflex coronary artery. Three weeks later, 3 x 10(8) human myoblasts carrying lac-z reporter gene were transplanted in multiple sites (0.25 mL each) into the left ventricular wall. Immunosuppression was achieved with 5 mg/kg cyclosporine for 6 weeks after cell transplantation. After animals were euthanized between 6 and 30 weeks after cell transplantation; the heart was removed for histological studies. Discontinuation of immunosuppression after 6 weeks of cell transplantation did not result in donor cell rejection. The lac-z-positive donor cells were detected in porcine host cardiac tissue for up to 30 weeks posttransplantation, expressing human skeletal myosin heavy chain. The results highlight the effectiveness of transient immunosuppression for myoblast transplantation for cardiac repair.