Combination of mass spectrometry and GST pull-down techniques to study potential interacting protein of PCV2 ORF4 / 生物工程学报

A novel protein encoded by the open reading frame 4 (ORF4) was recently discovered in porcine circovirus type 2 (PCV2). However, little is known about the interaction proteins of ORF4 which hindered better understanding the biological functions of ORF4 in the life cycle of PCV2. In the present study, the ORF4 was inserted into the multiple cloning site of pCMV-N-Flag-GST, yielding recombinant plasmid pCMV-N-Flag-GST-ORF4. The recombinant plasmid was transfected into 293T cells and the intracellular interaction complex of ORF4 were enriched and separated by GST pull-down and SDS-PAGE, sequentially. The potential interacting proteins of PCV2 ORF4 were stained with silver and identified by mass spectrometry (MS). Finally, five candidate ORF4-interacting proteins, including Serine/threonine-protein phosphatase 6 catalytic subunit, alpha cardiac muscle 1, actin, SEC14-like protein 5 and myosin 9 were identified. These results would benefit a better understanding of the biological function of ORF4 in PCV2 infected cells.

Clinicopathological features of ovarian Brenner tumors / 中华病理学杂志

Objective@#To investigate the clinicopathological characteristics and diagnosis of ovarian Brenner tumors.@*Methods@#Forty-seven cases of ovarian Brenner tumors were enrolled from January 2012 to May 2018 at Beijing Obstetrics and Gynecology Hospital, Capital Medical University. Clinical data, imaging examination, histopathological characteristics and immunohistochemical phenotype were analyzed.@*Results@#The age of the patients ranged from 30-73 years and the mean age was 55 years. Thirty-nine patients (83.0%) were postmenopausal. Forty cases (85.1%) of the Brenner tumors were benign, five (10.6%) borderline and two (4.3%) malignant. Usual tumor markers of ovarian carcinoma, including CA199 and CA125 were normal or mild elevated in the 47 cases. Imaging before surgery was not specific to Brenner tumors. Microscopically, benign Brenner tumors were composed of nests of bland, transitional-type cells within a fibromatous stroma. In our 5 cases of borderline Brenner tumors, mildly atypical transitional-type cells were projected into the cyst lumens and lack of stromal invasion. In 2 cases of malignant Brenner tumors, different degrees of nuclear atypial transitional-type cells exhibited stromal invasion. Immunohistochemical stains for CK7, GATA3, p63 and CK5/6 were positive in all cases. Ki-67 was less than 5% in Brenner tumors, and up to 20%-30% in malignant Brenner tumors.@*Conclusion@#Brenner tumors are mostly seen in postmenopausal patients and are usually benign. Imaging examination and usual ovarian tumor markers do not provide diagnostic value. Diagnosis and classification of Brenner tumors depend on histopathological evaluation.

Expression of type 1 and type 2 cytokines from serum of coal miners and the evaluation in surveillance of coal workers' pneumoconiosis at earlier stage / 中华预防医学杂志

Objectives@#To explore the expression regulation of type 1 and type 2 (Th1 and Th2) cytokines from serum of coal miners and the evaluation in surveillance of coal workers' pneumoconiosis, 630 coal miners were studied.@*Methods@#A total of 90 male patients diagnosed as coal workers' pneumoconiosis (CWP) in a institute for occupational health and 19 male workers newly diagnosed as CWP patients was chosen as CWP group with simple random sampling method from a coal mine group from January 2013 to December in 2015. 180 male coal miners with abnormal but not diagnosed as CWP were selected as CWP suspected group with simple random sampling methods, meanwhile 180 male coal miners with normal chest X-ray photograph was as dust-exposed group by 1∶1 matched as age. And 161 healthy males accepted pre-employed examination were selected as control group, CWP suspected group, dust-exposed group and control group called as non-CWP group. According to screening test and diagnosis test, the basic information and occupational history of all subjects were collected, and cytokines including IL-1β, IL-8, IFN-γ, IL-6 and IL-10 of serum were detected. Receiver operator characteristic (ROC) curve was used to determine the optimal cutoff value of each cytokine. Area under curve (AUC), the validity and reliability were calculated and judged.@*Results@#The average age of control group, dust-exposed group, CWP suspected group and CWP group were (27.4±5.0) , (43.4±10.7) , (48.2±6.2) , (64.7±7.0) years old, respectively. The median level of IL-1β, IL-8, IFN-γ and IL-6 in cases group (1 638.30, 2 099.49, 815.18,140.32 pg/ml) were higher than that of non-cases group (1 445.57, 1 402.26, 736.38, 95.73 pg/ml) (P<0.05) . The level of IL-8 (1 503.99 pg/ml) in CWP suspected group was higher than that of control group (1 295.67 pg/ml) and dust-exposed group (1 376.94 pg/ml) , but the level of IL-10 (654.08 pg/ml) was lower than that of control group (596.64 pg/ml) . The ratio of IFN-γ/IL-6 ranged from 5 to 8, and the ratio in CWP group (5.87) was lower than that of non-CWP group (7.61) . The IL-6 and IL-8 among the subjects of dust-exposed group in terms of the age distribution of among had reached statistical significance. According to ROC, the cutoff value of IL-1β, IL-6, IL-8, IL-10 and INF-γ reached 1 582.65, 116.53, 1 791.54, 581.08 and 792.69 pg/ml, respectively. The AUC was 0.668, 0.895, 0.859, 0.716 and 0.637, respectively. It was found that IL-6 and IL-8 could be used as biomarkers in detecting CWP, the sensitivity and specificity was 82.6% and 84.6%, 78.0% and 84.8%, respectively; Youden's index was 0.674 and 0.628 and the consistency rate was 84.3% and 83.7%, while Kappa value was 0.55 and 0.52.@*Conclusion@#There was Type 1 and type 2 cytokine dysregulation in CWP patients. IL-6 and IL-8 can be used as effective biomarkers to forecast lung injury before X-ray changes.

KRAS mutations analysis in mucinous epithelial lesions of the endometrium / 中华病理学杂志

Objective@#To investigate the frequency of KRAS mutation in mucinous epithelial lesions of the endometrium, and analyze the correlation between KRAS mutation and the clinicopathologic features.@*Methods@#The cohort included forty-three cases of mucinous epithelial lesions of the endometrium selected from July 2015 to October 2017 from Beijing Obstetrics and Gynecology Hospital, and 22 control cases. Genomic DNA was extracted from formalin-fixed paraffin-embedded tissue sections. Polymerase chain reaction amplification for KRAS exons 2 and 3 was performed, followed by sequencing using capillary electrophoresis. The Fisher exact test was used to compare the prevalence of KRAS mutation among the different groups.@*Results@#The patients′age ranged from 33 to 77 years [mean (55.12±9.34) years, median 55 years]. None of the eight cases of endometrial hyperplasia with mucinous differentiation without atypia showed KRAS mutation. The frequency of KRAS mutations was 1/10 in endometrial atypical hyperplasia, 1/12 in endometrioid carcinoma, 4/11 in endometrial atypical hyperplasia with mucinous differentiation (EAHMD), 6/15 in endometrioid carcinoma with mucinous differentiation (ECMD) and 8/9 in mucinous carcinoma (MC), respectively. The differences were statistically significant between MC versus EC (P<0.01) and MC versus ECMD (P<0.05).@*Conclusion@#The high frequency of KRAS mutation in EAHMD, ECMD and MC indicates that KRAS mutational activation is implicated in the pathogenesis of endometrial mucinous carcinoma.

High aldehyde dehydrogenase 1A1 (ALDH1A1) expression correlated with risk of lymph node metastasis in papillary thyroid carcinoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between ALDH1A1 expression and lymph node metastasis (LNM) in papillary thyroid carcinoma (PTC).</p><p><b>METHODS</b>One hundred and fifty-three paraffin-embedded specimens of PTC treated in the Beijing Tongren Hospital of Capital Medical University were selected from January 2006 to December 2013. The expression of ALDH1A1 was detected in both tumor tissues and adjacent non-tumor tissues by immunohistochemistry and several clinicopathological parameters (size, bilaterality, multifocality, tumor border and extrathyroidal extensions) were assessed by HE staining. The correlation of ALDH1A1 expression with LNM was analyzed.</p><p><b>RESULTS</b>In 153 cases of PTC, there were 82 cases with LNM, 126 cases with high ALDH1A1 expression in tumor tissues, and 112 cases with high ALDH1A1 expression in adjacent non-tumor tissues. On univariate analysis, patient age < 45 years, tumor size of 10 mm or more, invasive tumor border, and high ALDH1A1 expression in tumor tissues predicted LNM in PTC (P < 0.05), whereas gender, bilaterality, multifocality, extra-thyroidal extensions and high ALDH1A1 expression in adjacent non-tumor border did not (P > 0.05). On multivariate analysis, invasive tumor border, high ALDH1A1 expression in tumor tissues were found to be independent predictive factors for LNM in PTC (P < 0.05). After a follow-up of 42 months (median time), four patients developed locoregional recurrences, but no distance recurrence or disease related death were seen in 82 patients of follow up. The estimated 5-year locoregional recurrence was 4.88%. Of these four logcoregional recurrences, three involved lymph nodes and one involved the remaining thyroid. The ALDH1A1 expression in tumor tissues was high in all of recurrence cases.</p><p><b>CONCLUSION</b>High ALDH1A1 expression in tumor tissues is correlated with lymph node metastasis in PTC and may be used as an independent predictive factor of LNM, and may improve treatment and follow-up strategies for PTC.</p>

Urinary exfoliated cytology:a clinical pathological study of 735 cases / 临床与实验病理学杂志

Purpose To explore the pathological feature of urinary exfoliated cell examination and influence factors by retrospectively comparing the coincidence of diagnosis between urinary exfoliated cell examination and histopathologic results of cystoscopic biopsy. Methods 735 patients underwent both urinary exfoliated cell examination and histopathologic biopsy of cystoscope evaluation from No-vember 2010 to July 2014 in Peking University Shougang Hospital were enrolled in this study. The urinary exfoliated cells were treated with Pap staining, while the histopathologic biopsy were dealt with HE staining. All cases were divided into three groups according to the diagnosis of urinary exfoliated cell examination:negative group ( no cancer or atypical cell detected) , suspicious group ( atypical cell detected) and positive group ( cancer cell detected) . These above diagnoses were confirmed with the histopathologic biopsy. ROC curve analysis, Cochran-Armitage trend test and logistic regression analysis were performed to evaluate the sensitivity and the specificity of urinary exfoliated cell examination as well as the relationship between diagnoses with age and sex. Results The age range of 735 patients (551 male and 184 female) was 28 ~91 years and the median age was 69 years. There were 187 patients in the positive group, including 184 malignant and 3 false-positive cases. The suspicious group, including 186 cases, consisted of 67 malignant, 119 benign reactive changes. Of all 362 cases in the negative group, malignant tumor was detected in 90 cases. For histologic diagnosis, the AUC of ROC(95%CI)was 0. 800 (0. 767~0. 834), displaying significant difference as compared to the histological pathological diagnostic results(P<0. 001). As the cyto-histologic diagnostic level elevated from negative, suspicious to positive, the results of Co-chran-Armitage trend test showed significant differences(Z=15. 83, P<0. 001). If standardized with the histopathologic biopsy re-sults, the AUC (area under curve) of urinary exfoliated cell examination was 0. 800 (0. 767~0. 834) in ROC curve analysis was sig-nificantly larger (P<0. 001). Furthermore, we also found in Logistic regression that the incidence of cancer was 1. 04 (1. 03~1. 05) times higher if aged one year older ( P<0. 001 ) , while there was no significant relationship between the incidence and the sex ( P=0. 655). Conclusions The coincidence rate of urinary exfoliated cell examination increases with the malignant degree. A positive cor-relation is detected between age and the incidence of malignant tumor. Detailed clinical material can markedly improve the sensitivity and accuracy of cyto-histologic diagnosis.

The study of autophagy in alveolar macrophages of patients with coal workers' pneumoconiosis / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To evaluate the differences in the autophagy activity of alveolar macrophages between patients with different stages of coal workers' pneumoconiosis (CWP).</p><p><b>METHODS</b>A total of 116 coal workers were investigated in the field. Their lung lavage fluid was collected and purified to obtain alveolar macrophages. The morphological characteristics of autophagy were observed by transmission electron microscopy. The expression of autophagy marker (LC3) and autophagy regulators (Beclin1, mTOR, and p-mTOR) was measured by Western blot. The autophagy activity of alveolar macrophages was compared between dust-exposed subjects and patients with stage I, II, and III CWP.</p><p><b>RESULTS</b>The autophagy activity of alveolar macrophages differed between patients with different stages of CWP, according to transmission electron microscopy. Patients with stage II CWP had significantly higher protein expression of LC3 II/I and Beclin1 in pulmonary macrophages than those with stage ICWP (P < 0.05); patients with stage III CWP had significantly lower protein expression of LC3 II/I and Beclin1 in pulmonary macrophages than those with stage II CWP (P < 0.05), but had significantly higher protein expression of LC3 II/I and Beclin1 than those with stage I CWP (P < 0.05); patients with stage II CWP had a significantly higher protein expression of Beclin1 than the dust-exposed subjects (P < 0.05). Patients with stage II CWP had significantly lower expression of mTOR and p-mTOR in pulmonary macrophages than the dust-exposed subjects and those with stage I CWP (P < 0.05), while patients with stage III CWP had significantly higher expression of mTOR and p-mTOR than those with stage II CWP (P < 0.05).</p><p><b>CONCLUSION</b>The autophagy activity of alveolar macrophages varies between patients with different stages of CWP.</p>

Expression of microRNA-10a-5p in laryngeal epithelial premalignant lesions / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the expression of microRNA-10a-5p (miR-10a) in laryngeal epithelial premalignant lesions (LEPL) and to analyze the correlations of its dysregulation with clinicopathologic parameters of LEPL.</p><p><b>METHODS</b>According to the WHO classification (2005), 94 cases of LEPL were grouped into mild dysplasia (MID), moderate dysplasia (MOD), severe dysplasia (SD) and carcinoma in situ ( CIS). The expression of miR-10a in 94 cases of LEPL was detected by quantitative real-time polymerase chain reaction (qRT-PCR), and correlated with the clinical and follow-up data of all LEPL patients.</p><p><b>RESULTS</b>miR-10a was down-regulated in LEPL with increasing grade of dysplasia. There was significantly statistical difference between low-risk ( MID + MOD) and high-risk ( SD + CIS) lesion groups (P = 0.03). The linear regression analysis showed that miR-10a was correlated with grade and gender of LEPL (P < 0.05).</p><p><b>CONCLUSIONS</b>Down regulation of miR-10a may be a useful molecular marker for grading of LEPL and diagnosis of early laryngeal cancer.</p>

Comparison of long-term and short-term mild hypothermia on severe traumatic brain injury:a systematic review / 医学研究生学报

Objective Mild hypothermia provides protection for the brain and improves prognosis in the treatment of severe traumatic brain injury, which is widely acknowledged.The article aimed to analysis the prognosis and complications of long-term and short-term mild hypothermia on patients with severe traumatic brain injury. Methods According to the cochrane systematic review methods, thorough search was carried out in databases including Cochrane, Pubmed, Embase, CBM, CNKI, Wanfang and VIP.Eval-uation was made on the quality of selected documents, and Revman5.2 software was applied for meta analysis after data extraction. Results Long-term mild hypothermia achieved improved prognosis compared to short-term mild hypothermia ( GOS score 4 ~5 ) [RR=1.37, 95%CI (1.14, 1.64), P=0.0006].No significant difference was found between these two therapies in pneumonia in-cidence (P=0.94), arrhythmia incidence ( P=0.54) and stress ulcer incidence (P=0.99). Conclusion In comparison to short-term hypothermia therapy, long-term mild hypothermia therapy improved the prognosis of patients with severe traumatic brain inju-ry without obvious increase in the incidence of pneumonia, arrhythmia and stress ulcer.

Investigation on psychological stress of mine emergency rescue personnel / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate psychological stress and influence factors on the mine emergency rescue personnel.</p><p><b>METHOD</b>564 mine emergency rescue personnel from a rescue group were select as subjects, and 60 designers from a steel design institute were as controls. Self-made questionnaire and general job stress questionnaire were used to investigate the basic information, rescue history, psychosomatic symptoms, depression symptoms, daily job stress and negative emotions of emergency rescue personnel. SPSS17.0 software was used to analysis the psychological stress on the mine rescue personnel and its influence factors.</p><p><b>RESULTS</b>The detection rate (41.94%) of depression symptoms in rescue team was higher than that of controls (24.90%). The score of daily job stress was higher than that of logistical support. The older age group with higher negative emotional and daily job stress than the younger. The highest negative emotion was in age group of more than 40 years old. The highest score of daily job stress was in ≤30 years old. The score of depression and psychosomatic symptoms were higher than those of the college and the above. The scores of depression in group of duration of rescue <10 years was higher than that of duration≥10 years. The score of daily job stress is the lowest in rescue for 1 to 2 times per year and the highest in group of simulation training once a week. The score of daily job stress and depressive symptoms were getting higher with the extension of combat duty time. Age, hours of combat duty, training times a week, education and life events were the main affecting fectors on mental health of mine rescuers.</p><p><b>CONCLUSIONS</b>Mine rescuers have more psychological stress than generic population. The psychological stress of the mine crew is related to age, education, life events, training and combat readiness duty time.</p>

Relationship between genetic polymorphisms of matrix metalloproteinase-2 and -3 and susceptibility to silicosis / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the relationship between the genetic polymorphisms of matrix metalloproteinase-2 (MMP-2) (-735) and matrix metalloproteinase-3 (MMP-3) (-1171) and the susceptibility to silicosis.</p><p><b>METHODS</b>A case-control study was conducted in 113 patients diagnosed with stage I silicosis (case group) and 115 dust-exposed workers without silicosis (control group); the two groups had the same sex, ethnic group, and type of dust and similar age and cumulative exposure time. Polymerase chain reaction-restriction fragment length polymorphism was used to determine the genotypes of MMP-2 (-735) and MMP-3 (-1171).</p><p><b>RESULTS</b>No significant difference was observed in age, cumulative exposure time, or smoking rate between cases and controls (P > 0.05). The frequencies of genotypes C/C, C/T, and T/T at MMP-2 C-735T in the case group were 57.5% (65/113), 31.0% (35/113), and 11.5% (13/113), respectively, which were significantly different from those of the control group (69.6% (80/115), 26.9% (31/115), and 3.5% (4/115)), χ² = 6.542, P < 0.05). The frequencies of T allele in cases and controls were 27.0% and 17.0%, respectively, which were significantly different from each other χ² = 6.704, P < 0.05). Carriage of T allele at MMP-2 C-735T increased the risk of silicosis (OR = 1.811, 95% CI: 1.151-2.847). The frequencies of genotypes 6A/6A, 5A/6A, and 5A/5A at MMP-3 A-1171A were 67.2% (76/113), 24.8% (28/113), and 8.0% (9/113), respectively, in the case group, versus 59.1% (68/115), 37.4% (43/115), and 3.5% (4/115) in the control group (χ² = 5.519, P > 0.05).</p><p><b>CONCLUSION</b>Genetic polymorphism at MMP-2 C-735T is significantly associated with the development of silicosis. Carriage of T allele increases the risk of silicosis among workers exposed to dust. No significant association was found between MMP-3 A-1171A polymorphism and silicosis in this study.</p>

AN EXPERIMENTAL STUDY ON CHRONIC SPORTS INJURIES IN QUADRICEPS AND CALF MUSCLES AND THEIR INSERTION AREAS / 中国运动医学杂志

The experiment was designed to simulate active training with animal model. A dynamic and static trai-ning program for adult rats was carried out 30-60'/day, lasted from 25 to 232 days. The rats were killedperiodical1y in batches and the sections were prepared for exploring histological changes in knee and ankle areas on L/M. Degenerative cbanges in quadriceps and gastrocanemius muscles appeated by degrees aftet a period of in-tensive training. The nuclei proliferated in chains or clusters arranged in the center and the edge of thefibers. The number of nuclei for each chain or cluster could be as much as 20 or 30. Meanwhile, therewere also ischemic changes in small vessels. All these were the characteristic changes induced by mictotraumain muscles under repeated active and strenous training. In calcaneal and patellar ateas, there were changes of cartilaginous degeneration, necrosis and prolifera-tion (cartilaginous exostoses and osteophytes) too. The typical changes of enthesiopatby---degeneration, necrosis and proliferation (osteophyte) could beobserved in calcaneal tendon and its insertion area and in superior insertion atea of lower patella. The occur-rence of enthesiopathy in this experiment migbt be related to tke stress of traction force within muscle andcartilage during running in upward treadmill.

THE PROTECTIVE EFFECT OF TAURINE ON ALCOHOLIC LIVER DAMAGE IN RAT / 营养学报

Objective: To observe the preventive effect of taurine on alcoholic liver damage in rat.Methods: The model was induced by filling stomach with alcohol, and the treatment groups were given two different concentrations of taurine, and the control group was given 0.9%NS, then we measured the blood triglycerides (TG), and the pathological changes of liver quantitatively.Results: Two different le-vels of taurine can significantly decrease the level of TG and ameliorate the liver damage of rats (P0.05), and the 5% taurine group can also decrease the mean arterial pressure (P