A high resolution reaction microscope with universal two-region time-focusing method.

This paper presents a novel reaction microscope designed for ion-atom collision investigations, established at the Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou, China. Its time-of-flight (TOF) spectrometer employs an innovative flight-time focusing method consisting of two acceleration regions, providing optimal time focusing conditions for charged fragments with diverse initial velocities. The TOF spectrometer's axis intentionally tilts by 12° relative to the ion beam direction, preventing potential obstructions from the TOF grid electrodes. The introduced focusing method allows for a flexible time-focusing TOF spectrometer design without restricting the length ratio of the two regions. In addition, this configuration in our case significantly suppresses noise on the recoil ion detector produced by residual gas in the ion beam trajectory, which is a considerable challenge in longitudinal spectrometers. In a test experiment on the single electron capture reaction involving 62.5 keV/u He2+ ions and a helium atomic beam, the recoil longitudinal momentum resolution achieved 0.068 atomic units. This novel configuration and successful test run show excellent precision for ion-atom collision studies.

[Clinical research of multisystem inflammatory syndrome in children].

Objective: To analyze the clinical characteristics of children with multisystem inflammatory syndrome (MIS-C) associated with SARS-CoV-2 in China, and to improve the understanding of MIS-C among pediatricians. Methods: Case series study.Collect the clinical characteristics, auxiliary examinations, treatment decisions, and prognosis of 64 patients with MIS-C from 9 hospitals in China from December 2022 to June 2023. Results: Among the 64 MIS-C patients, 36 were boys and 28 were girls, with an onset age being 2.8 (0.3, 14.0) years. All patients suffered from fever, elevated inflammatory indicators, and multiple system involvement. Forty-three patients (67%) were involved in more than 3 systems simultaneously, including skin mucosa 60 cases (94%), blood system 52 cases (89%), circulatory system 54 cases (84%), digestive system 48 cases (75%), and nervous system 24 cases (37%). Common mucocutaneous lesions included rash 54 cases (84%) and conjunctival congestion and (or) lip flushing 45 cases (70%). Hematological abnormalities consisted of coagulation dysfunction 48 cases (75%), thrombocytopenia 9 cases (14%), and lymphopenia 8 cases (13%). Cardiovascular lesions mainly affected cardiac function, of which 11 patients (17%) were accompanied by hypotension or shock, and 7 patients (12%) had coronary artery dilatation.Thirty-six patients (56%) had gastrointestinal symptoms, 23 patients (36%) had neurological symptoms. Forty-five patients (70%) received the initial treatment of intravenous immunoglobulin in combination with glucocorticoids, 5 patients (8%) received the methylprednisolone pulse therapy and 2 patients (3%) treated with biological agents, 7 patients with coronary artery dilation all returned to normal within 6 months. Conclusions: MIS-C patients are mainly characterized by fever, high inflammatory response, and multiple organ damage. The preferred initial treatment is intravenous immunoglobulin combined with glucocorticoids. All patients have a good prognosis.

Two- and three-body dissociations of C3H6 isomer dications investigated by 4 keV/u Ar8+ impact.

The fragmentation dynamics of two isomers of C3H6, cyclopropane and propene, induced by 4 keV/u Ar8+ are investigated employing a reaction microscope. Four two-body and two three-body dissociation channels of C3H6 2+ dications are identified for each isomer, among which the channels involving CC bond breaking are found to be much more favored than H3 + and H2 + formation channels. The observation of the CH3 + or H3 + formation channels from cyclopropane are direct evidence of the proton migration within the carbon skeleton before dissociation. Obvious isomer effects are revealed by comparing the relative branching ratios of different channels of the two isomers. Moreover, it was shown that a sequential dissociation mechanism with H elimination prior to CC bond cleavage may be dominant for the two three-body dissociation channels for both isomers.

[Studies of the norm of Karitane Parenting Confidence Scale(KPCS)among parents of infants in urban areas of China].

Objective: To establish the norm of the Chinese version of Karitane Parenting Confidence Scale (KPCS) in urban areas of China. Methods: From August to December 2017, the parents of 2 216 children (<36 months old) were selected from 15 cities (Beijing, Lianyungang, Hangzhou, Chengdu, Xi'an, Guangzhou, Changsha, Jinan, Guiyang, Ningbo, Dalian, Qinhuangdao, Maanshan, Chongqing and Wuhan) in 14 provinces by stratified random sampling. The general demographic characteristics and parents' parenting confidence were collected by a self-made questionnaire and KPCS Chinese version. The percentile norm was established. P3, P10 and P25 were used as the criteria to define the degree of lack of parenting confidence. Results: The age of mothers was (30.67±4.29). The age of the father was (32.50±4.99) years old. There were 726 (32.76%), 759 (34.25%) and 731 (32.99%) infants in 6-12, 12-23 and 24-35 months old groups. The total scores of P50, P25, P10 and P3 of KPCS (Chinese version) of infant parents in urban areas in China were 41, 38, 33, and 29 respectively. When the scores of parents were 34-37, 30-33, and ≤ 29, they were judged as mild, moderate, and severe lack of parenting confidence. There was no significant difference in the Chinese version of KPCS between parents of different age groups and parents of different gender (χ²=3.53, P=0.171; χ²=1.41, P=0.236). Each factor score≤P3 is defined as the boundary score, and the corresponding boundary scores of "parenting" "support" and "competence" were 13, 9, and 5 respectively. Conclusion: The Chinese version of KPCS can be used to assess the parenting confidence of infants in urban areas of China. It can used as one of the bases for scientific and objective evaluation of the parenting status of families.

[Clinical effect evaluation of high risk human papilloma virus E6/E7 mRNA in triaging women with atypical squamous cells of undetermined significance].

Objective: To evaluate the performance of high risk human papilloma virus (HR-HPV) E6/E7 mRNA in triaging women with atypical squamous cells of undetermined signification (ASCUS). Methods: The ASCUS cohort determined by liquid-based cytology test in the cervical cancer screening queue in Luoshan County, Xinyang City, Henan Province 2017 were selected. The population underwent colposcopy biopsy and pathological tests, combined with HPV16 or 18 (HPV16/18), HR-HPV DNA and HPV E6/E7 mRNA test. By using the pathological result as the gold standard, the sensitivity, specificity, positive predictor (PPV), negative prediction (NPV), referral rate and 95% confident interval (CI) of HPV E6/E7 mRNA, HR-HPV DNA, HPV16/18 were calculated, respectively. Results: The average age of 312 ASCUS women was 52.6±7.3 years old. Among the 290 women diagnosed as normal by pathology, 64 (22.1%) were HPV E6/E7 mRNA positive, 86 (29.7%) were HR-HPV DNA positive, and 19 (6.6%) were HPV16/18 positive. All of the cervical intraepithelial neoplasia (CIN) cases determined by pathology were both HPV E6/E7 mRNA and HR-HPV DNA positive. The sensitivity, specificity, PPV, NPV and referral rate of HPV E6/E7 mRNA for predicting CIN2+ lesion in women with ASCUS were 100.0% (95%CI: 72.3, 100.0), 77.8% (95%CI: 72.8, 82.1), 13.0% (95%CI: 7.2, 22.3), 100.0% (95%CI: 98.4, 100.0) and 24.7%. Compared with HPV E6/E7 mRNA, the sensitivity of HR-HPV DNA was similar with HPV E6/E7 mRNA, but with a lower specificity [70.2% (95%CI: 64.8, 75.1), P<0.05], a higher referral rate (32.1%, P<0.05); while HPV 16/18 had a high specificity (93.4%, 95%CI: 90.0, 95.7) and a low sensitivity (30.0%, 95%CI: 10.8, 60.3). Based on the age stratification by age 45, the sensitivity of HPV E6/E7 mRNA in both age groups was 100.0%. The specificity of HPV E6/E7 mRNA in group of ≥45 years was a little higher than that in group of <45 years [79.0% (95%CI: 73.7, 83.5) versus 68.3% (95% CI: 53.0, 80.4)], but the difference was not statistically significant (P>0.05). Conclusions: The ASCUS woman triaging effect of HPV E6/E7 mRNA detection is better than those of HR-HPV and HPV16/18 under certain conditions. The application of HPV E6/E7 mRNA detection to triage ASCUS population can avoid unnecessary colposcopy referral and reduce the missed diagnosis of cervical lesions.

[Performance of high-risk HPV typing test in early diagnosis of cervical cancer].

Objective: To evaluate the performance of High-risk HPV typing detection in cervical cancer screening. Methods: A total of 3 231 women were recruited as the subjects of cervical cancer screening from Jiyuan city of Henan provinces from June to July 2017. All women underwent HPV DNA test. The women with cytological examination ≥ASCUS or cytological examination negative and HPV 16/18 positive underwent colposcopy biopsy and pathological examination. Pathological diagnosis was used as the gold standard. Sensitivity, specificity, positive predictive value (PPV), negative predictive value(NPV) as well as corresponding 95% confidence interval (CI) of HR-HPV and HPV 16/18 were calculated. Results: The mean age of 3 231 subjects selected in this study was 46.84±10.00 (21-64) years old. 524 subjects had the positive results of HR-HPV, including 91 of HPV16 and 15 of HPV18. Pathological test result CIN2+ was the gold standard. The sensitivity and specificity of HR-HPV for cervical precancer lesions screening were 93.75 (95%CI: 79.85-98.27) and 84.56% (95%CI: 83.26-85.77), respectively. To compared with HR-HPV, HPV16/18 had low sensitivity (65.63%, 95%CI: 48.31-79.59)and higher specificity (97.44%, 95%CI: 96.83-97.93). After age stratification by age 30 and 45, the sensitivity of HPV 16/18 was same with HR-HPV (100%, 95%CI: 34.24-100.00), the specificity of HPV 16/18 was higher than HR-HPV (98.71%, 95%CI: 96.27-99.56 vs 84.48%, 95%CI: 79.27-88.58) in<30 age group.The sensitivity of HR-HPVin 30-45 and ≥45 age group were higher than HPV16/18 (85.71%, 95%CI: 48.69-97.43 vs 71.43%, 95%CI: 35.89-91.78, 95.65%, 95%CI: 79.01-99.23 vs 60.87%, 95%CI: 40.79-77.84), but the specificity werelower than HPV16/18 (86.89%, 95%CI: 84.58-88.90 vs 98.51%, 95%CI: 97.51-99.11、83.49%, 95%CI: 81.81-85.04 vs 96.80%, 95%CI: 95.94-97.48). Conclusions: HR-HPV detection has relatively high sensitivity and specificity in cervical cancer screening. For >30 years old women, HR-HPV is more recommended in cervical cancer screening. Therefore, HR-HPV detection is an effective method for cervical cancer screening.

[Evaluation the triage performance of p16/Ki-67 dual staining for HR-HPV positive women].

Objective: This study aimed to evaluate the clinical performance of p16/Ki-67 dual staining for triage high risk HPV (HR-HPV) infected women. Method: Target objects were women who infected HR-HPV and received colposcopy examination between April and December of 2016 at the Second Affiliated Hospital of Zhengzhou University. Gynecologists collected the cervical exfoliated cells from eligible women for p16/Ki-67 dual staining, LBC testing and HPV DNA testing. Histology diagnosis were used as gold standard. Sensitivities, specificities, positive predictive values (PPVs), negative predictive values (NPVs) of p16/Ki-67 dual staining, LBC testing and HPV16/18 testing for triage of HR-HPV positive population were calculated and compared. Results: A total of 295 HR-HPV infected women were selected, and the mean age was (44.29±11.48) years old. Positive rates of p16/Ki-67 dual staining, HPV16/18 testing and LBC testing were 70.17% (207), 56.95% (168) and 85.76% (253), respectively. When CIN2+as the endpoint, among the three triage methods, sensitivity of p16/Ki-67 dual staining was 90.00% (95%CI: 85.06%-93.43%), higher than the value of HPV 16/18 testing, but lower than the value of LBC testing. Specificity, PPV and NPV of p16/Ki-67 dual staining were the highest [71.58% (95%CI: 61.81%-79.67%), 86.96% (95%CI:81.69%-90.88%) and 77.27% (95%CI: 67.49%-84.78%)]. When detection for CIN3+, sensitivity of p16/Ki-67 dual staining was 92.90% (95%CI: 87.74%-95.99%), lower than the value of LBC testing, but higher than the value of HPV16/18 testing. Specificity of p16/Ki-67 dual staining was 55.00% (95%CI: 46.74%-63.00%), lower than the value of HPV16/18 testing, but higher than the value of LBC testing. PPV of p16/Ki-67 dual staining was 69.57% (95%CI: 62.99%-75.43%), lower than the value of HPV 16/18 testing, but higher than the value of LBC testing. NPV of p16/Ki-67 dual staining was 87.50% (95%CI: 78.99%-92.87%), higher than value of HPV 16/18 testing, but lower than the value of LBC testing. Conclusion: p16/Ki-67 dual staining has better clinical effects than HPV 16/18 testing and LBC testing for triage women with HR-HPV infection.

Correlation between endometrial receptivity with expressions of IL-1 and VEGF in rats with polycystic ovary syndrome.

OBJECTIVE: To investigate the correlation between endometrial receptivity with expressions of interleukin-1 (IL-1) and vascular endothelial growth factor (VEGF) in rats with polycystic ovary syndrome (PCOS). MATERIALS AND METHODS: A total of 24 female Sprague-Dawley (SD) 21 days old were randomly divided into control group (n=12) and PCOS group (n=12). Rats in the control group were normally raised, and PCOS model was established in rats of the PCOS group. All the rats were sacrificed when they grew to 80 days old. Immunohistochemistry was applied to detect the expressions of IL-1 and VEGF. Western blotting was performed to measure the relative expressions of IL-1 and VEGF proteins. Quantitative Polymerase Chain Reaction (qPCR) was utilized to determine the relative messenger ribonucleic acid (mRNA) expressions of IL-1 and VEGF. Data related to endometrial receptivity were detected. RESULTS: The expression levels of IL-1 and VEGF in the PCOS group declined markedly compared with those in the control group, and the differences were statistically significant (p<0.05). PCOS group had notably lower protein expressions of IL-1 and VEGF than the control group, with statistically significant differences (p<0.05). The mRNA expressions of IL-1 and VEGF in PCOS group were significantly lower than those in the control group, displaying statistically significant differences (p<0.05). Compared with that in the control group, the endometrial receptivity of rats in the PCOS group was reduced evidently, and the difference was statistically significant (p<0.05). Both IL-1 and VEGF had positive correlations with the endometrial receptivity. CONCLUSIONS: Both IL-1 and VEGF are positively correlated with the endometrial receptivity in the case of PCOS, which can serve as therapeutic targets for PCOS and improve endometrial receptivity in the future.

[Practice of parenting and related factors on children aged 0-5 in the urban areas of China].

Objective: To characterize the relations between the practice of parenting and associated factors on children (0-5 years old) in urban areas of China, in order to provide evidence for promoting the early development of children and to provide positive guidance and service programs on parenting. Methods: A total of 4 515 parents from 15 cities (14 provinces) were surveyed with a self-administered questionnaire. Parenting and Family Adjustment Scales (PAFAS) was used, including parameters as: consistency and coercive parenting, positive encouragement, parent-child relationship and parental emotion adjustment, family relationship and parental teamwork aspects, etc. Both single factor analysis and multiple linear regression were used to examine the associations between parenting practice, individual, parental and family factors. Results: The mean score of PAFAS was 21.00 (15.00-28.00), associated with factors as children's age, only-child family, premature delivery, father's education level, confidence on parenting, problems regarding the parental mood, annual family income, family structure and behavior on seeking professional help, etc. Results showed that there were big differences on the practice of parenting in China and influenced by variety of factors. Conclusions: The general situation of parenting was well, in the urban areas of China. The practice of parenting was associated with a series of individual, parental and family factors. Programs on improving the parenting skills and promoting the early development of children, should be highlighted.

Downregulation of hsa_circ_0000285 serves as a prognostic biomarker for bladder cancer and is involved in cisplatin resistance.

Bladder cancer remains a very challenging disease to treat with the high rates of recurrence and progression associated with current therapies. Although the association between bladder cancer pathology and circRNAs remains undetermined, circRNAs signatures may be useful as prognostic and predictive factors and clinical tools for assessing disease state, treatment response and outcome. This study investigates if these circRNAs can be used as biomarkers for bladder cancer diagnosis and predicting treatment response. Herein, qPCR measured the expression of hsa_circRNA_100783, hsa_circ_0000285 and hsa_circRNA_100782 in bladder cancer tissues. It was established that sa_circ_0000285, but not hsa_circRNA_100782 and hsa_circRNA_10078, are significantly reduced in bladder cancer tissues and serum compared to adjacent tissues and healthy controls. Moreover, hsa_circ_0000285 expression was lower in cisplatin-resistant bladder cancer patients than in those who were cisplatin-sensitive. Here, hsa_circ_0000285 was associated with tumor size (p<0.001), differentiation (p<0.001), lymph node metastasis (p=0.038), distant metastasis (p=0.004) and TNM stage (p=0.013). Further analysis showed that hsa_circ_0000285 would be an independent prognostic factor for bladder cancer patient outcome. In conclusion, our study indicates hsa_circ_0000285 may be a novel biomarker for bladder cancer because of its involvement in bladder cancer chemo-sensitivity.

[The clinical performance of high risk HPV DNA test among triage women with cytology showing atypical squamous cells of undetermined significance].

Objective: To evaluate the performance of high risk human papillomavirus (HR-HPV) DNA test for triage ASC-US in Chinese population. Methods: The study population was from Jiyuan city in Henan Province where a cervical cancer screening cohort (4 026 women) was set up from April to July in 2017. Women with ASC-US and complete recall information of colposcope were selected as the study objects. Self-designed questionnaire was used to survey the general information and medical history of cervical cancer, thencervical cytological speciments were collected for cytodiagnosis and HPV DNA test, followed by colposcopy, lesion biopsy and histology diagnose. The diagnosis of histology was used as the gold standard. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) as well as corresponding 95% confidential interval (CI) were calculated. Results: The average age of the included 393 women with ASC-US were (50.81±9.22) years old. The positivity of high risk human papillomavirus (HR-HPV), HPV16 and HPV18 were 35.6%, 10.69% and 2.80%, respectively. The sensitivity, specificity, PPV and NPV for HR-HPV to detect CIN2+ were 84.38% (68.25%-93.14%), 68.70% (63.74%-73.26%), 19.29% (13.61%-26.61%) and 98.02% (95.46%-99.15%). When compared with HR-HPV, HPV16/18 had a lower sensitivity (59.38% (42.26%-74.48%)) and a higher specificity(91.14% (87.75%-93.65%)), the PPV and NPV were 59.38% (25.32%-50.98%) and 91.14% (93.61%-97.77%). Conclusion: It was more effective for HR-HPV to triage ASC-US than HPV16/18. In order to improve the effect of HPV16/18, it should include more HR-HPV types which were more prevalent in Chinese population.

[Association between epicardial fat volume and coronary artery calcification in patients with chronic kidney disease].

Objective: To evaluate whether epicardial fat volume (EFV) is related to coronary artery calcification in patients with chronic kidney disease(CKD). Method: Multi-slice computed tomography was performed in 30 healthy subjects and 120 patients with CKD. Cross-sectional tomographic cardiac slices from base to apex were traced semi-automatically using a Volume Viewer of AW4.3 off-line workstation, and EFV was measured by assigning Hounsfield units ranging from -250 to -30 HU to fat.The coronary artery calcification score was assessed by CaScoring software. High density lipoprotein cholesterol(HDL-C), low density lipoprotein cholesterol(LDL-C) and collecting the body mass index (BMI), dialysis route, history of diabetes and coronary artery disease were used to analyze the relationship between EFV and other risk factors in patients with CKD. Results: There were 60.8%(73/120) male (mean age 62.8 years) and 39.2%(47/120) female (mean age 66.6 years) in the patients cohort, 73.3%(88/120) patients had coronary artery disease, 55.8%(67/120) had diabetes, 21 patients were on peritoneal dialysis and 9 on hemodialysis. EFV was apparently higher in stage 4-5 D CKD group compared with the control group((140.03±54.71), (145.01±64.56)and (141.45±62.04) cm(3) vs.(92.42±39.56)cm(3), P=0.007, 0.015 and 0.001), was similar between CKD3 and control group, and EFV was significantly higher in peritoneal dialysis group than in hemodialysis group and in coronary artery disease group compared with no coronary artery disease group((140.67±70.31) cm(3) vs.(105.22±61.49) cm(3), P=0.002). EFV was obviously higher in diabetes group than no diabetes group((148.41±65.78) cm(3) vs.(110.53±62.37) cm(3), P=0.007). CACS was apparently increased in stage 3-5 CKD group compared with the control group(140.0 vs.4.3, P<0.001). (3)When the patients were divided into four groups according to the eGFR, EFV was positively associated with CACS(r(s)=0.539, P=0.004) in control group, and the association become more robust in patients with CKD5(r(s)=0.841, P<0.000 1). EFV was related to age(r=0.662, P=0.005), BMI(r=0.648, P=0.009)and HDL-C(r=-0.433, P=0.024), but not related to eGFR and LDL-C. EFV was related to CACS(r=0.427, R(2)=0.182 3, P<0.001). CACS was positively correlated to age and BMI (all P<0.05)and negatively correlated with eGFR(P<0.05). Conclusions: Measurement of EFV may provide another useful noninvasive indicator of coronary artery calcification in CKD patients.

[A retrospective clinical study of sixty-three cases with persistent inflammation immunosuppression and catabolism syndrome].

Objective: To study the clinical characteristics and prognosis of patients with persistent inflammation immunosuppression and catabolism syndrome (PICS) in ICU. Methods: A total of 126 patients admitted to ICU (ICU stay of more than 10 days, age≥18 years) between January 2014 to December 2014 were retrospectively studied.Data were collected from electronic medical records including demographics, underlying disease, Acute Physiology and Chronic Health Evaluation Ⅱ (APACHEⅡ) score, Sequential Organ Failure Assessment (SOFA) score, laboratory parameters, ICU acquired infections and clinical outcome. Results: The overall incidence of PICS in ICU patients (ICU stay of more than 10 days) was 50.0%(63/126). There were no significant differences in baseline data such as gender, age, APACHEⅡscore, SOFA score and underlying diseases between the two groups(all P>0.05). Compared with the non-PICS group, there were more patients with gastrointestinal perforation in the PICS group (P=0.042), however, the medical or surgical admission did not differ between the two groups(P>0.05). During the stay in ICU, the PICS group showed a higher risk of developing acquired infections compared with the non-PICS group[PICS 63.5%(40/63) vs non-PICS 23.8%(15/63); P<0.001]. The infections were more caused by Candida in the PICS group than the non-PICS group [PICS 22.4%(11/49) vs non-PICS 2/17; P=0.003]. Moreover, the PICS group experienced longer stay in ICU[PICS(31.6±28.8) days vs non-PICS (20.4±11.3) days; P=0.0046] and higher ICU mortality [PICS 28.6%(18/63) vs non-PICS 6.3%(4/63), P=0.001]. Conclusion: PICS is a common manifestation of patients who stay in ICU more than 10 days, which is associated with high risk of ICU acquired infections, prolonged length of stay and high mortality.

Effect of miRNA-203 on cervical cancer cells and its underlying mechanism.

miRNA-203 is involved in the development and progression of various types of cancer. However, its role in cervical cancer remains unclear. The aim of this study was to investigate the effect of miRNA-203 on the proliferation and migration of HeLa cervical cancer cells, as well as survivin expression in these cells. A miRNA-203 primer probe was designed according to a sequence obtained from NCBI. The expression of miRNA-203 in cervical epithelial cells and cervical cancer cells was detected by quantitative reverse transcriptase-polymerase chain reaction. The miRNA-203 expression pattern was compared between these two cell lines. The cervical cancer cells were transfected with miRNA-203 mimic or inhibitor to determine their effects on proliferation and migration. The expression of the miRNA-203 target protein (survivin) was analyzed by western blot. Cervical cancer cells showed reduced miRNA-203 expression compared to cervical epithelial cells. Transfection of miRNA-203 mimic upregulated the expression of miRNA-203, suppressed cell proliferation and migration, and downregulated survivin expression (P < 0.05). However, downregulation of miRNA-203 expression did not affect proliferation, migration, and survivin expression in cervical cancer cells (P > 0.05). In conclusion, upregulation of miRNA-203 in cervical cancer cells inhibits the proliferative and migratory capacities of these cells by downregulating the expression of survivin.

Sensitive and rapid detection of Pectobacterium atrosepticum by targeting the gyrB gene using a real-time loop-mediated isothermal amplification assay.

UNLABELLED: This study reports the development of a real-time, loop-mediated isothermal amplification (RealAmp) assay for the detection of Pectobacterium atrosepticum (P. atrosepticum). A phylogenetic tree was constructed based on the gyrB gene of P. atrosepticum and related species. Pectobacterium atrosepticum from different sources can be clustered in the same branch with 100% support rate. The RealAmp primers targeting the gyrB gene of P. atrosepticum worked most efficiently at 61·0°C. Compared with 55 related bacterial strains, the eight P. atrosepticum strains displayed positive reaction in the RealAmp assay. The melting temperature (Tm) of P. atrosepticum amplified products was about 85·0°C. The detection limit of the RealAmp assay for the detection of P. atrosepticum in pure culture was approx. 3 CFU reaction(-1) . The detection limit of the RealAmp assay for the detection of P. atrosepticum in artificially contaminated samples was 22 CFU reaction(-1) . The detection rate of the RealAmp assay for the detection of potato tubers was 28·5-32·0% higher than that of the conventional PCR. In summary, a specific, sensitive and rapid RealAmp assay based on the gyrB gene of P. atrosepticum, which can be easily performed and real-time monitored, was established. SIGNIFICANCE AND IMPACT OF THE STUDY: Potato blackleg caused by Pectobacterium atrosepticum (P. atrosepticum) which is mainly transmitted through the seed potato leads to the decline in potato production. To reduce yield loss, rapid detection of P. atrosepticum in seed potato remains essential. Based on the gyrB gene of P. atrosepticum, species-specific primers were designed. A real-time, loop-mediated isothermal amplification (RealAmp) assay was established for the detection of P. atrosepticum. The RealAmp assay is a specific, rapid and sensitive method for P. atrosepticum detection. Therefore, it provides an effective diagnosis of potato blackleg in both the growing and stored potato.

Temperature dependence of photoluminescence spectra for green light emission from InGaN/GaN multiple wells.

Three green light emitting InGaN/GaN multiple quantum well (MQW) structures with different In composition grown by metal-organic chemical vapor deposition are investigated by the X-ray diffraction and the temperature-dependent photoluminescence (PL) measurements. It is found that when the In composition increases in the InGaN/GaN MQWs, the PL spectral bandwidth may anomalously decrease with increasing temperature. The reduction of PL spectral bandwidth may be ascribed to the enhanced non-radiative recombination process which may lower the light emission efficiency of the localized luminescent centers with shallow localization energy in the high-In-content InGaN quantum wells and also cause a reduction of integrated PL intensity.

Antimicrobial activity of recombinant mature bovine neutrophil ß-defensin 4 on mycobacterial infection.

BACKGROUND: Previous studies have shown that human cathelicidin and defensins have effective antimicrobial activity against Mycobacterium spp. OBJECTIVE: To investigate the antimycobacterial effect of mature bovine neutrophil ß-defensin (mBNBD) 4 against Mycobacterium spp. infection for the first time. DESIGN: mBNBD4 protein was expressed in Pichia pastoris GS115. We used immunofluorescent assay to detect whether the recombinant mBNBD4 had entered the macrophages. The antimycobacterial activity of mBNBD4 was tested through colony-forming unit (cfu) assay. Morphological changes in the cell wall of M. bovis treated with mBNBD4 were observed by scanning electron microscope. RESULTS: mBNBD4 was expressed and successfully purified from P. pastoris with intact antimicrobial activity. The recombinant protein was able to enter Raw 264.7 macrophages and exhibited potent in vitro bactericidal activity against M. smegmatis and M. bovis. The cell wall of M. bovis was disrupted after interaction with mBNBD4. Exogenous addition of mBNBD4 to both Raw 264.7 and THP-1 derived macrophages reduced the intracellular survival of M. bovis and M. tuberculosis relative to control cells. CONCLUSION: Our data show that mBNBD4 plays an important role in inhibiting mycobacterial growth and in controlling intracellular survival of mycobacteria. mBNBD4 could therefore an effective antimycobacterial molecule in combination with other measures.

MTHFR C677T variant reduces risk of sporadic Parkinson's disease in ethnic Chinese.

BACKGROUND: Genetic variability of methylenetetrahydrofolate reductase (MTHFR) may be associated with Parkinson's disease (PD). Its role in ethnic Chinese population is still unclear. Our study aimed to investigate whether MTHFR C677T variation was linked to PD risk in a Han Chinese population from mainland China. METHODS: To investigate the association with the risk of PD, we analyzed the single-nucleotide polymorphism C677T in MTHFR gene using a case-control methodology. A total of 1482 subjects included 765 patients with idiopathic PD, and 717 age- and sex-matched controls were recruited in this study. RESULTS: The T allele of MTHFR C677T was associated with a decreased risk of PD (OR = 0.80, 95% CI: 0.688-0.926, P = 0.003). Patients with CT + TT genotypes have a decreased risk of PD compared with those with CC genotypes (OR = 0.66, 95%CI: 0.532-0.813, P = 0.000). CT + TT subjects cannot be differentiated from CC subjects based on their clinical features. CONCLUSION: We showed that the C677T polymorphism in MTHFR gene was associated with decreased PD susceptibility in a Han Chinese population from mainland China. Efforts to fully elucidate the pathophysiologic role of the variant in PD should be necessary.

Validation of a simple universal IELISA for the diagnosis of human brucellosis.

The definitive diagnosis of brucellosis requires isolation of the agent, although negative isolation does not rule out the infection. In contrast, serological testing is more sensitive and, therefore, preferred in clinical practice. The majority of reported cases around the world were caused by Brucella melitensis, B. abortus, B. suis and B. canis. The first three species contain O-polysaccharide (OPS) on the cell surface, but B. canis contains no measurable OPS on the rough lipopolysaccharide (R-LPS). A universal indirect enzyme immunoassay for the detection of serum antibody to smooth and rough Brucella spp. in both normal (u-IELISA®) and rapid forms (R-u-IELISA®) has been developed, and, therefore, the potential use of this method was assessed in comparison to cELISA, conventional tests, IELISA and RSAT on a total of 478 sera. The 77 sera from blood donors with no clinical or epidemiological evidence of brucellosis and negative serological tests showed a specificity of 100 % for both u-IELISA® and R-u-IELISA®, with a cut-off value of %P 24 and %P 18, respectively. Sera from 49 culture-positive cases (16 B. suis, 15 B. abortus, 12 B. melitensis and 6 B. canis) yielded a sensitivity of 98 % for u-IELISA® and 95.9 % for R-u-IELISA®. In general, u-IELISA® showed good correlation with cELISA and IELISA for the detection of antibodies to smooth and rough Brucella strains, as well as for monitoring patients during treatment, but R-u-IELISA® seems to need additional optimisation. u-IELISA® is simple to perform and could be a suitable test for field laboratories and hospitals lacking skilled personnel.