[HbA1c comparison and diagnostic efficacy analysis of multi center different glycosylated hemoglobin detection systems].

Objective: Compare and analyze the results of the domestic Lanyi AH600 glycated hemoglobin analyzer and other different detection systems to understand the comparability of the detection results of different detectors, and establish the best cut point of Lanyi AH600 determination of haemoglobin A1c (HbA1c) in the diagnosis of diabetes. Methods: Multi center cohort study was adopted. The clinical laboratory departments of 18 medical institutions independently collected test samples from their respective hospitals from March to April 2022, and independently completed comparative analysis of the evaluated instrument (Lanyi AH600) and the reference instrument HbA1c. The reference instruments include four different brands of glycosylated hemoglobin meters, including Arkray, Bio-Rad, DOSOH, and Huizhong. Scatter plot was used to calculate the correlation between the results of different detection systems, and the regression equation was calculated. The consistency analysis between the results of different detection systems was evaluated by Bland Altman method. Consistency judgment principles: (1) When the 95% limits of agreement (95% LoA) of the measurement difference was within 0.4% HbA1c and the measurement score was≥80 points, the comparison consistency was good; (2) When the measurement difference of 95% LoA exceeded 0.4% HbA1c, and the measurement score was≥80 points, the comparison consistency was relatively good; (3) The measurement score was less than 80 points, the comparison consistency was poor. The difference between the results of different detection systems was tested by paired sample T test or Wilcoxon paired sign rank sum test; The best cut-off point of diabetes was analyzed by receiver operating characteristic curve (ROC). Results: The correlation coefficient R2 of results between Lanyi AH600 and the reference instrument in 16 hospitals is≥0.99; The Bland Altman consistency analysis showed that the difference of 95% LoA in Nanjing Maternity and Child Health Care Hospital in Jiangsu Province (reference instrument: Arkray HA8180) was -0.486%-0.325%, and the measurement score was 94.6 points (473/500); The difference of 95% LoA in the Tibetan Traditional Medical Hospital of TAR (reference instrument: Bio-Rad Variant II) was -0.727%-0.612%, and the measurement score was 89.8 points; The difference of 95% LoA in the People's Hospital of Chongqing Liang Jiang New Area (reference instrument: Huizhong MQ-2000PT) was -0.231%-0.461%, and the measurement score was 96.6 points; The difference of 95% LoA in the Taihe Hospital of traditional Chinese Medicine in Anhui Province (reference instrument: Huizhong MQ-2000PT) was -0.469%-0.479%, and the measurement score was 91.9 points. The other 14 hospitals, Lanyi AH600, were compared with 4 reference instrument brands, the difference of 95% LoA was less than 0.4% HbA1c, and the scores were all greater than 95 points. The results of paired sample T test or Wilcoxon paired sign rank sum test showed that there was no statistically significant difference between Lanyi AH600 and the reference instrument Arkray HA8180 (Z=1.665,P=0.096), with no statistical difference. The mean difference between the measured values of the two instruments was 0.004%. The comparison data of Lanyi AH600 and the reference instrument of all other institutions had significant differences (all P<0.001), however, it was necessary to consider whether it was within the clinical acceptable range in combination with the results of the Bland-Altman consistency analysis. The ROC curve of HbA1c detected by Lanyi AH600 in 985 patients with diabetes and 3 423 patients with non-diabetes was analyzed, the area under curve (AUC) was 0.877, the standard error was 0.007, and the 95% confidence interval 95%CI was (0.864, 0.891), which was statistically significant (P<0.001). The maximum value of Youden index was 0.634, and the corresponding HbA1c cut point was 6.235%. The sensitivity and specificity of diabetes diagnosis were 76.2% and 87.2%, respectively. Conclusion: Among the hospitals and instruments currently included in this study, among these four hospitals included Nanjing Maternity and Child Health Care Hospital in Jiangsu Province (reference instrument: Arkray HA8180), Tibetan Traditional Medical Hospital of TAR (reference instrument: Bio-Rad Variant Ⅱ), the People's Hospital of Chongqing Liang Jiang New Area (reference instrument: Huizhong MQ-2000PT), and the Taihe Hospital of traditional Chinese Medicine in Anhui Province (reference instrument: Huizhong MQ-2000PT), the comparison between Lanyi AH600 and the reference instruments showed relatively good consistency, while the other 14 hospitals involved four different brands of reference instruments: Arkray, Bio-Rad, DOSOH, and Huizhong, Lanyi AH600 had good consistency with its comparison. The best cut point of the domestic Lanyi AH600 for detecting HbA1c in the diagnosis of diabetes is 6.235%.

Developmental changes of free amino acids in amniotic, allantoic fluids and yolk of broiler embryo.

1. A study was conducted to evaluate the developmental changes of protein and free amino acid concentrations in amniotic, allantoic fluids and yolk during the incubation period of broiler eggs.2. A total of 120 Cobb 500 fertile eggs were individually weighed and then placed in an incubator. On incubation day: 8, 11, 13, 14, 16, and/or 18, amniotic, allantoic fluids and yolk were collected from 20 eggs for analysis of protein content and free α-amino acid concentration in allantoic and amniotic fluids and yolk.3. The total protein concentration in amniotic fluid increased from d 11 of incubation, and reached a peak at d 16 (69.85 g/l; P < 0.01), then declined at d 18 (P < 0.05). The total protein concentration in allantoic fluid increased with age of the embryo (P < 0.01). Crude protein concentration in yolk decreased (P < 0.05) from d 0 to 8, then increased gradually from d 8, and reached a peak at d 16 (P < 0.05). The concentration of most free amino acids in amniotic and allantoic fluids and yolk was related to embryo weight. Amniotic fluid amino acids gradually increased from d 13 to 18, with arginine being the most abundant at d 11 and 14. Glutamate was the most predominant amniotic fluid amino acid at d 16 and 18. From d 13 to 18, the concentrations of most α-amino acids in allantoic fluid increased, and reached a peak at d 18 (aspartate, 373 µmol/l; asparagine, 519 µmol/l; glutamine, 1230 µmol/l; threonine, 537 µmol/l; citrulline, 112 µmol/l; arginine, 2747 µmol/l; alanine, 276 µmol/l; tyrosine, 330 µmol/l; tryptophan, 212 µmol/l; valine, 140 µmol/l; phenylalanine, 102 µmol/l; isoleucine, 92.39 µmol/l; lysine, 1088 µmol/l; P < 0.05). Glutamine was the second most abundant amino acid in allantoic fluid at d 13 and 18. Glutamate was the most abundant α-amino acids at d 8, and 13 in the yolk.4. These results demonstrated that the concentration of free α-amino acids in chicken embryo fluid was related to embryo weight. Arginine, glutamine and glutamate were abundant free α-amino acids in chicken embryo fluid, to support the higher rates of tissue protein synthesis and growth for the embryo.

[Comparison of clinical features between sporadic pancreatic neuroendocrine tumors and those associated with multiple endocrine neoplasia type 1].

Objective: To compare the clinical features of multiple endocrine adenoma type 1 (MEN-1) associated pancreatic neuroendocrine neoplasms (pNENs) as well as sporadic pNENs. Methods: The clinical data of 28 sporadic pNENs patients and 10 MEN-1-related pNENs patients admitted to the First Affiliated Hospital of Nanjing Medical University from January 2010 to June 2021 were collected. Meanwhile, by searching PubMed database and reviewing the clinical data of 20 foreign patients with MEN-1-related pNENs which were reported at the same time.Compare and analyze the similarities and differences between MEN1-associated pNENs and sporadic pNENs in clinical features, such as family history, blood tests, pathological diagnostic indicators, tumor grade, stage and metastasis, treatment and prognosis and so on. Results: A total of 58 pNENs patients were included, and there were 30 MEN1-related pNENs patients and 28 sporadic pNENs patients. Eighteen patients (60%) had a family history of MEN1-related pNENs, and the mean age of onset was (35.3±13.0)years. There were no patients (0) with family history of sporadic pNENs, and the mean age of onset was(55.3±13.4)years. In contrast, the differences in family history, age of onset and NSE were statistically significant(all P<0.05).Among the pathological diagnostic indicators, there were 19 patients (63.3%) with Grade G2 of MEN1-related pNENs, and 25 patients (83.3%) with somatostatin receptor 2(SSTR2) negative. In sporadic pNENs, there were 16 patients (57.1%) with Grade G2 and 9 patients (32.1%) with SSTR2 negative. The differences in pathological grade, immunohistochemistry (Chromogranin A, CD56, and somatostatin receptor 2, SSTR2) between the two groups were statistically significant(all P<0.05). In terms of tumor staging and metastasis, 21 patients with MEN-1-related pNENs had metastasis (70%) and 20 patients with stage Ⅰ and Ⅱ AJCC (71%) in all. Eight patients with sporadic pNENs had metastasis (26.7%) and 8 patients were with stage Ⅰ and Ⅱ AJCC (28.6%). By contrast, the differences in total metastasis rate, AJCC stage and distant metastasis between the two groups were statistically significant(all P<0.05). In terms of treatment and prognosis, there was no statistical significance in the differences between surgical treatment and prognosis (P>0.05), and the difference was also not statistically significant in survival rate between them (P>0.05). Conclusions: There are no significant differences between MEN1-related pNENs and sporadic pNENs in terms of treatment, prognosis, and survival rate, but there are significant differences in clinical features, pathological features and the staging and grading of tumors. The rate of tumor grade, stage and metastasis of sporadic pNENs is higher.

[Application of combined detection of multiple pathogens in diagnosis and treatment of coronavirus disease 2019].

Due to the wide variety of pathogens causing respiratory tract infection and the close symptoms, coronavirus disease 2019 (COVID-19) needs to be differentiated from other common infections. Early comprehensive detection and accurate identification of respiratory infection pathogens is of great value for early diagnosis, curative effect, as well as monitor of the diseases. Combined detection of multiple pathogens can quickly and accurately detect and distinguish the pathogens, then provide rapid and reliable laboratory diagnostic basis for further treatment. This article elaborates the application and development of multiplex detection assay in the diagnosis of COVID-19 according to the recent research.

[Cardiovascular disease risk in diabetes patients aged 40 years old and above in China].

Objective: To evaluate the incidence of cardiovascular disease (CVD), ischemic and hemorrhagic cardiovascular events among Chinese diabetic patients aged 40 years and above with different CVD risk levels. Methods: This study enrolled participants aged 40 years and above in 15 provinces from a prospective cohort study, the China-PAR project (Prediction for Atherosclerotic Cardiovascular Disease Risk in China). Participants were categorized into two groups according to the presence or absence of diabetes at baseline. Individuals were further classified into low (0-4.9%), moderate (5%-9.9%) and high risk groups (≥10%), based on predicted ten-year CVD risk using the China-PAR equations. Two followed-up surveys were conducted between 2007 and 2015 to identify CVD events, which were defined as nonfatal acute myocardial infarction, or death due to coronary heart disease, or stroke. Ischemic cardiovascular events included nonfatal acute myocardial infarction, or death due to coronary heart disease, or ischemic stroke. Hemorrhagic cardiovascular events included subarachnoid hemorrhage and intracerebral hemorrhage. The incidences of CVD, ischemic and hemorrhagic cardiovascular events were compared in diabetes and non-diabetes population with different CVD risk levels. Results: This study included 89 209 participants aged 40 years and above, the average follow-up period was 8.5 years. The age was (54.8±9.4) years, and 36 794 (41.2%) were men, and 5 730 (6.4%) were diabetic patients. In diabetes patients aged 40 years and above, 53.7% (3 075/5 730) were at high risk of CVD. Age-and sex-adjusted incidence of CVD, ischemic and hemorrhagic cardiovascular events (1 066.93/100 000 person-years, 824.23/100 000 person-years, and 211.56/100 000 person-years) were significantly lower in diabetes patients than those in non-diabetes population with high CVD risk (1 773.73/100 000 person-years, 1 228.18/100 000 person-years, and 446.49/100 000 person-years) (all P<0.001). Among high CVD risk populations, incidence of ischemic events was significantly higher in diabetic patients than in non-diabetes population (1 638.47/100 000 person-years vs. 1 228.18/100 000 person-years, P<0.001), but incidence of hemorrhagic events tended to be lower in diabetic patients than in non-diabetes population (415.70/100 000 person-years vs. 446.49/100 000 person-years, P=0.635). Incidence of ischemic and hemorrhagic events were similar between diabetes patients and non-diabetes population at low or moderate CVD risk groups (all P>0.05). Conclusions: More than half of diabetes patients aged 40 years and above in China have high CVD risk. The incidence of CVD, ischemic and hemorrhagic cardiovascular events are different in diabetic patients with different CVD risk levels.

Efficient high-power dual-wavelength lime-green Nd:YVO4 lasers.

An efficient high-power dual-wavelength lime-green Nd:YVO4 self-Raman laser is inventively developed by using two different lithium triborate (LBO) crystals. The first and second LBO crystals are employed to generate the 559 nm lime and 532 nm green lasers, respectively. The temperature of the first LBO crystal is fixed at the optimal phase matching, whereas the temperature of the second LBO crystal is tuned to flexibly control the relative strengths between the 532 and 559 nm waves. When the temperature of the second LBO crystal is set for the maximal total output power, the 532 and 559 nm output powers, respectively, are 7.1 and 2.9 W at a pump power of 31.6 W, corresponding to a conversion efficiency of 31.6%. When the temperature is controlled for the balanced output, the 532 and 559 nm powers, respectively, are 4.3 and 4.2 W at a pump power of 31.6 W, corresponding to a conversion efficiency of 26.9%.

[Consistency of ALK Ventana-D5F3 immunohistochemistry interpretation in lung adenocarcinoma among Chinese histopathologists].

Objective: To understand the consistency of ALK Ventana-D5F3 immunohistochemistry (IHC) interpretation in Chinese lung adenocarcinoma among histopathologists from different hospitals, and to recommend solution for the problems found during the interpretation of ALK IHC in real world, with the aim of the precise selection of patients who can benefit from ALK targeted therapy. Methods: This was a multicenter and retrospective study. A total of 109 lung adenocarcinoma cases with ALK Ventana-D5F3 IHC staining were collected from 31 lung cancer centers in RATICAL research group from January to June in 2018. All cases were scanned into digital imaging with Ventana iSCANcoreo Digital Slide Scanning System and scored by 31 histopathologists from different centers according to ALK binary (positive or negative) interpretation based on its manufacturer's protocol. The cases with high inconsistency rate were further analyzed using FISH/RT-PCR/NGS. Results: There were 49 ALK positive cases and 60 ALK negative cases, confirmed by re-evaluation by the specialist panel. Two cases (No. 2302 and No.2701) scored as positive by local hospitals were rescored as negative, and were confirmed to be negative by RT-PCR/FISH/NGS. The false interpretation rate of these two cases was 58.1% (18/31) and 48.4% (15/31), respectively. Six out of 31 (19.4%) pathologists got 100% accuracy. The minimum consistency between every two pathologists was 75.8%.At least one pathologist gave negative judgement (false negative) or positive judgement (false positive) in the 49 positive or 60 negative cases, accounted for 26.5% (13/49), 41.7% (25/60), respectively, with at least one uncertainty interpretation accounted for 31.2% (34/109). Conclusion: There are certain heterogeneities and misclassifications in the real world interpretation of ALK-D5F3 IHC test, which need to be guided by the oncoming expert consensus based on the real world data.

Accuracy and reliability of digital craniofacial measurements using a small-format, handheld 3D camera.

OBJECTIVES: Craniofacial assessments often involve three-dimensional facial imaging using an expensive camera with 6 SLR lenses to analyse the positions and relations of anatomic landmarks. Recently, a 3D small-format, handheld camera was developed; however, the accuracy and reliability of this system are largely unknown. The purpose of this study was to evaluate the accuracy and reliability of this system. MATERIALS & METHODS: A total of 30 sets of evaluations were completed by 2 examiners on 5 human subjects, using 3 different methods: direct callipers, 3D handheld camera and conventional tripod 3D camera images. Each evaluation included 29 anthropometric landmarks that were used as reference points for facial analysis. Two examiners marked the landmarks directly on the faces and measured linear distances using the 3 measurement methods. RESULTS: Accuracy analysis was performed for handheld vs direct calliper vs conventional camera measurements. Each of these analyses yielded a grand mean of correlation coefficients of .98. Bias measurements revealed that the handheld and conventional camera methods yielded larger measurements than direct callipers (with a mean difference of 1.74, 1.56 mm, respectively, for rater 1 and 0.94, 1.02 mm, respectively, for rater 2). When compared to one another, both the handheld camera and the conventional camera methods yielded similar values for most measurements, with the average overall difference between these modalities of 0.03 mm for rater 1 and 0.07 mm for rater 2. CONCLUSIONS: The 3D handheld camera showed high accuracy and reliability in comparison with traditional models, indicating that this system may provide a useful tool in craniofacial anthropometry.

[The association of fruit and vegetable intake with the changes of serum lipid levels in middle-aged and older Chinese population].

Objective: To investigate the association of fruit and vegetable intake with long-term changes of serum lipid levels in middle-aged Chinese and older Chinese population. Methods: The study analyzed the data collected in the 2004 and 2007-2008 cohorts of China Multicenter Collaborative Study of Cardiovascular Epidemiology. Finally, 4 495 participants from 10 groups in various regions of China, who both were followed up in the two cohorts, were included for data analysis. They were aged 41-66 years in 2004. Fruit and vegetable consumption were collected with a simple food frequency questionnaire. The percent changes (Δ%) of serum TC, TG, HDL-C and LDL-C between the two surveys were calculated. Multivariate linear regression models were used to estimate the association of fruit and vegetable intake in 2004 with percentage changes of serum lipid levels during the two surveys. Results: The proportions of individuals who consumed fruits <250, 250-499 and ≥500 g/week were 24.0%, 21.8% and 54.2%, respectively. The proportions of individuals who consumed vegetables <500 and ≥500 g/day were 76.7% and 23.3%, respectively. Compared with fruit intake <250 g/week, the regression coefficients (95% CI) of ΔTC% in 250-499 and ≥500 g/week group were -1.54 (-2.71, -0.37) and -1.77 (-2.79, -0.76). And the regression coefficients (95% CI) of ΔLDL-C% were -2.43 (-4.39, -0.48) and -2.89 (-4.59, -1.19). Compared with vegetable intake <500 g/d, the regression coefficients (95%CI) of ΔTC% and ΔLDL-C% in vegetable intake ≥500 g/d group were -1.01 (-1.95, -0.06) and -1.83 (-3.41, -0.24). However, fruit and vegetable intake had no relationship with ΔTG% and ΔHDL-C%. Conclusion: The consumption of fruit and vegetable was inversely related to long-term changes of TC and LDL-C in middle-aged and older population, but no association was found for changes of TG and HDL-C.

[The relationship between physical activity and incident hypertension in rural Chinese].

Objective: To investigate the relationship between physical activity (PA) and the risk of incident hypertension among population in rural areas of China. Methods: The Community Intervention of Metabolic Syndrome in China & Chinese Family Health Study (CIMIC) was conducted in 2007-2008. Data on PA, smoking, drinking, blood pressure and other variables were obtained at baseline. Then the follow-up study of incident hypertension was performed during 2012-2015. A total of 41 457 participants aged ≥18 years and free from hypertension at baseline were included in the final analyses. PA was calculated as metabolic equivalent (MET) for each participant. Cox proportional hazard models were used to explore the relationship of PA with incident hypertension according to the quartiles of PA. Results: A total of 6 780 participants developed hypertension during an average follow up of 5.8 years. The annual incidence of hypertension was 2.80%. Compared to participants in the first quartile of PA, HR (95%CI) of incident hypertension decreased with the level of PA of 0.92 (0.86, 0.99), 0.72 (0.67, 0.77) and 0.70 (0.65, 0.75) for the 2(nd), 3(rd) and 4(th) quartile, respectively (P(trend)<0.001). In subgroup analyses, compared to the first quartile, hazards of hypertension among normotensive participants (systolic blood pressure less than 120 mmHg (1 mmHg=0.133 kPa) and diastolic blood pressure less than 80 mm Hg) in the 2(nd), 3(rd) and 4(th) quartile were 0.82 (0.70, 0.95), 0.73 (0.63, 0.85) and 0.78 (0.67, 0.90), respectively (P(trend)=0.002). Among participants with prehypertension (systolic blood pressure from 120 to 139 mmHg and/or diastolic blood pressure from 80 to 89 mmHg), similar trend for the relationship of PA and incident hypertension was also found with HR (95%CI) of 0.94 (0.87, 1.01), 0.71 (0.65, 0.77) and 0.66 (0.61, 0.71) for the 2(nd), 3(rd) and 4(th) quartile, respectively (P(trend)<0.001). Conclusion: There was linear trend association between PA and incident hypertension. Increased PA in daily life may be a protective factor against hypertension.

[Relationship of lipid accumulation product with hypertension and diabetes among Beijing residents study].

Objective: To explore the lipid accumulation product (LAP) formula for Beijing adults and to investigate the relationship between lipid accumulation product and hypertension, as well as diabetes. Methods: A cross-sectional study with a representative sample of 19 606 residents in Beijing aged 18-79 years was conducted in 2011. The sex-specific hypothetical minimum waist circumference (WC) was calculated in order to obtain the more applicable LAP formula. Multivariate logistic regression was used to analyze the associations of LAP, combination of LAP and body mass index (BMI) with hypertension and diabetes. Results: The LAP formula for Beijing adults was established as follows: LAP (male)= (WC-61.3) × TG, and LAP (female)= (WC-55.6) × TG. There was an obvious trend of increased risk of hypertension and diabetes with an increment in the tertiles of the LAP both in men and women. The OR (95%CI) for hypertension in the group with highest tertile LAP was 3.62 (3.11-4.22) in men, and 5.79 (4.84-6.93) in women, compared with the lowest tertile group, respectively; and the corresponding OR for diabetes was 3.47 (2.73-4.41) in men, and 4.10 (2.90-5.80) in women, respectively. Compared with the lowest tertile of LAP and normal BMI group, the OR (95%CI) for hypertension in the highest tertile of LAP and obesity group was 6.79 (5.50-8.37) in men, and 9.75 (7.76-12.25) in women, respectively; while the corresponding value for diabetes was 3.97 (2.87-5.49) in men, and 4.13 (2.78-6.14) in women, respectively. Conclusion: The elevated level of LAP was associated with an increased risk of hypertension and diabetes among Beijing adults. LAP could be an important predictor for hypertension and diabetes.

[Neovascularization aggravated hepatic fibrosis in combined hypoxia NASH rat model].

Objective: To optimize the construction of combined hypoxia NASH rat model on the basis of preliminary work, and to explore the role of neovascularization in the process of hepatic fibrosis. Methods: 32 rats were divided randomly to four groups that were null control group(A group ), hypoxia group(B group), high fat diet group(C group ) and high fat diet plus hypoxia group (D group ),treated with null , Intraperitoneal injection of NaNO(2), high fat diet and high fat diet plus Intraperitoneal injection of NaNO(2) respectively. Every group was observed for 16 weeks, B and D group was treated with Intraperitoneal injection of NaNO(2) 20 mg/kg.d at the laster 8 weeks. Liver histology NASH activity score(NAS) and Fibro score(FibroS), biochemical index were detected in this combined hypoxia NASH rat model(D group), meanwhile the changes of HIF1α, inflammatory factor and neovascularization were measured by ELISA, realtime PCR and immunohistochemistry. Results: Liver tissue NAS > 4 was seen in C and D group. D group showed NASH characteristics, including significantly steatosis at liver acinar 3 area(mostly a microvesicular type fat droplets mixed with macrovesicular type), hepatocyte balloon degeneration, obvious lobular inflammation, while fibrosis score increased significantly, including visible hepatic sinusoid fibrosis, fibrosis around portal vein, and bridging fibrosis in a considerable portion of the rats. Compared with C group, biochemical indicators of aspartate aminotransferase (AST), HIF1α, neovascularization-related VEGFA, VEGFR2 mRNA level increased obviously and the expression of immunohistochemistry VEGFR2, CD34 enhanced markedly in D group(p < 0.05). Conclusion: A combined hypoxia NASH rat model can be established throught feeding 16 weeks' high-fat diet then intraperitoneal injection of NaNO(2) 20 mg / kg.d at the laster 8 weeks, meanwhile chronic hypoxia can accelerate this combined hypoxia NASH model liver fibrosis process. In this process neovascularization promoted the formation of hepatic fibrosis in this model.

Myostatin mRNA expression and its association with body weight and carcass traits in Yunnan Wuding chicken.

Myostatin (MSTN) is expressed in the myotome and developing skeletal muscles, and acts to regulate the number of muscle fibers. Wuding chicken large body, developed muscle, high disease resistance, and tender, delicious meat, and are not selected for fast growth. Broiler chickens (Avian broiler) are selected for fast growth and have a large body size and high muscle mass. Here, 240 one-day-old chickens (120 Wuding chickens and 120 broilers) were examined. Twenty chickens from each breed were sacrificed at days 1, 30, 60, 90, 120, and 150. Breast and leg muscle samples were collected within 20 min of sacrifice to investigate the effects of MSTN gene expression on growth performance and carcass traits. Body weight, carcass traits, and skeletal muscle mass in Wuding chickens were significantly (P < 0.05) lower than those in broiler chickens at all time points. Breast muscle MSTN mRNA was lower in Wuding chickens than in broilers before day 30 (P < 0.05). After day 30, breast muscle MSTN expression was higher in Wuding chicken than in broilers (P < 0.05). Leg muscle MSTN mRNA expression was higher in Wuding chicken than in broilers at all ages except for day 60 (P < 0.05). Correlation analysis revealed that breast muscle MSTN expression has a greater effect in slow growing Wuding chickens than in the fast growing broilers. In contract, leg muscle MSTN mRNA level has a greater effect in broilers than in Wuding chickens. MSTN regulates growth performance and carcass traits in chickens.

Comparison and analysis of Wuding and avian chicken skeletal muscle satellite cells.

Chicken skeletal muscle satellite cells are located between the basement membrane and the sarcolemma of mature muscle fibers. Avian broilers have been genetically selected based on their high growth velocity and large muscle mass. The Wuding chicken is a famous local chicken in Yunnan Province that undergoes non-selection breeding and is slow growing. In this study, we aimed to explore differences in the proliferation and differentiation properties of satellite cells isolated from the two chicken breeds. Using immunofluorescence, hematoxylin-eosin staining and real-time polymerase chain reaction analysis, we analyzed the in vitro characteristics of proliferating and differentiating satellite cells isolated from the two chicken breeds. The growth curve of satellite cells was S-shaped, and cells from Wuding chickens entered the logarithmic phase and plateau phase 1 day later than those from Avian chicken. The results also showed that the two skeletal muscle satellite cell lines were positive for Pax7, MyoD and IGF-1. The expression of Pax7 followed a downward trend, whereas that of MyoD and IGF-1 first increased and subsequently decreased in cells isolated from the two chickens. These data indicated that the skeletal muscle satellite cells of Avian chicken grow and differentiate faster than did those of Wuding chickens. We suggest that the methods of breeding selection applied to these breeds regulate the characteristics of skeletal muscle satellite cells to influence muscle growth.

Association between loop diuretic use and fracture risk.

SUMMARY: Loop diuretic use has been shown to be associated with an increased fracture risk, but the findings have been inconsistent. The present meta-analysis suggests that loop diuretics show a significant positive association with the overall risk of total fractures and, specifically, hip fractures. INTRODUCTION: Despite being widely used, there is limited, prospective randomized trial evidence regarding the skeletal effects of loop diuretics. Previous observational studies have reported conflicting findings regarding the association between loop diuretic use and the risk of fractures. METHODS: This meta-analysis of observational studies assessed the association between loop diuretic use and the risk of fractures. The PubMed, EMBASE, and OVID databases were searched for prospective cohort and case-control studies. Relative risks (RR) with 95% confidence intervals (CI) were derived using random-effects models throughout the analysis. RESULTS: Thirteen studies (4 cohort studies and 9 case-control studies) were included, involving 842,644 participants and 108,247 fracture cases. Compared with non-users, people who had taken loop diuretics had an approximately 15% higher risk of total fractures (95% CI, 1.04-1.26; p<0.01), with high heterogeneity between studies (I2=80.5%; p<0.01). The RR was 1.14 (95% CI, 1.08-1.19) for hip fractures and 0.99 (95% CI, 0.93-1.05) for lower arm or wrist fractures. The RR was 1.05 (95% CI, 1.00-1.11) in prospective cohort studies and 1.22 (95% CI, 1.00-1.44) in case-control studies. There was no evidence of publication bias. CONCLUSION: The results suggest that loop diuretics show a significant positive association with the overall risk of total fractures and hip fractures.

A non-PCR SPR platform using RNase H to detect MicroRNA 29a-3p from throat swabs of human subjects with influenza A virus H1N1 infection.

As in all RNA viruses, influenza viruses change and mutate constantly because their RNA polymerase has no proofreading ability. This poses a serious threat to public health nowadays. In addition, traditional pathogen-based detection methods may not be able to report an infection from an unknown type or a subtype of virus if its nucleotide sequence is not known. Because of these factors, targeting host microRNA signatures may be an alternative to classify infections and distinguish types of pathogens as microRNAs are produced in humans shortly after infection. Although this approach is in its infant stage, there is an urgent need to develop a rapid reporter assay for microRNA for disease control and prevention. As a proof of concept, we report herein for the first time a non-PCR MARS (MicroRNA-RNase-SPR) assay to detect the microRNA miR-29a-3p from human subjects infected with influenza virus H1N1 by surface plasmon resonance (SPR). In our MARS assay, RNase H is employed to specifically hydrolyze the RNA probes immobilized on the gold surface where they hybridize with its cognate target cDNAs miR-29a-3p, where it was formed from reverse transcription with mature miR-29a-3p specific stem-looped primers. After the digestion of the RNA probe by RNase H, the intact cDNA was released from the RNA-DNA hybrid and bound to a new RNA probe for another enzymatic reaction cycle to amplify signals. With assay optimization, the detection limit of our MARS assay for miR-29a-3p was found to be 1 nM, and this new assay could be completed within 1 hour without thermal cycling. This non-PCR assay with high selectivity for mature microRNA provides a new platform for rapid disease diagnosis, quarantine and disease control.

Implicating the H63D polymorphism in the HFE gene in increased incidence of solid cancers: a meta-analysis.

A number of previous studies have demonstrated that the HFE H63D polymorphism is associated with increased risk of incidence multiple types of cancer, including colorectal cancer, breast cancer, liver cancer, pancreatic cancer, and gynecological malignant tumors. However, the clinical outcomes were inconsistent. Therefore, this meta-analysis was conducted to summarize the effect of the H63D variant on the incidence of solid tumor. PubMed and EMBASE databases were searched for articles associating the HFE H63D polymorphism with cancer risk. The relationships were evaluated by calculating the pooled odds ratios (ORs) with 95% confidence intervals (CIs). A total of 28 studies, including 7728 cancer cases and 11,895 controls, were identified. Statistically significant associations were identified between the HFE H63D polymorphism and solid cancer risk (CG vs CC, OR = 1.14, 95%CI = 1.07-1.23, P < 0.001; GG vs CC, OR = 1.28, 95%CI = 1.06-1.55, P = 0.010; CG/GG vs CC, OR = 1.16, 95%CI = 1.08-1.24, P < 0.001; GG vs CC/CG, OR = 1.24, 95%CI = 1.02-1.49, P = 0.027). In the subgroup analysis, we illustrated the effect of the H63D polymorphism on hepatocellular carcinoma and pancreatic cancer risk, particularly in the Asian and African subgroups; however, this was not observed in gynecological malignant tumors. In summary, this analysis provided strong evidence that the HFE H63D polymorphism may play a critical role in the increased aggressiveness of hepatocellular carcinoma and pancreatic cancer.

Inhibitory effects of spironolactone on myocardial fibrosis in spontaneously hypertensive rats.

This study evaluated the inhibitory effects of spironolac-tone, a non-selective aldosterone receptor antagonist, on hypertension-induced myocardial fibrosis. Collagen I and III contents was detected in the myocardial tissue of spontaneously hypertensive rats (SHRs) after spironolactone administration. Twenty male SHRs were assigned to the spironolactone group or control group (N = 10 each); 7 Wistar-Kyoto rats (WKY) were also used. Spironolactone dissolved in ddH2O was administered via gavage at a dosage of 20 mg·kg(-1)·day(-1). Meanwhile, the control and WKY groups were administered equivalent volumes of ddH2O for 16 weeks. Western blotting was used to detect the contents of collagen I in myocardial tissue; observations were performed using polarizing microscopy, and the area integration and ratio of collagen I/III were subsequently calculated. Compared to the WKY group, col-lagen I synthesis was significantly higher in the control group (1.87 ± 0.2 vs 1.21 ± 0.7, P < 0.05). After 16 weeks of treatment, collagen I contents were significantly lower in the spironolactone group than in the control group (1.42 ± 0.05 vs 1.87 ± 0.2, P < 0.05). The ar-eas of collagen I and collagen I/III ratio were significantly smaller in the spironolactone group than in the control group (6400 ± 259 vs 12,019 ± 734 pixels, 15.64 ± 1.34 vs 20.8 ± 3.04 pixels, respec-tively; P < 0.05). However, there were no significant differences in the area of collagen III among the three groups. In conclusion, spi-ronolactone improves myocardial collagen deposition, preventing myocardial fibrosis in SHRs.

Clinical significance of SHMT1 rs1979277 polymorphism in Asian solid tumors: evidence from a meta-analysis.

Published data regarding the association between the cytosolic serine hydroxymethyltransferase (SHMT1) C1420T (Leu474Phe) polymorphism and solid tumor risk have shown inconclusive results. To derive a more precise estimation of the relationship, we performed a meta-analysis of 23 published studies that included 14,409 cancer cases and 16,996 controls. A comprehensive search was conducted to identify all eligible studies of the SHMT1 rs1979277 polymorphism and solid tumor risk. The pooled odds ratios (ORs) and the 95% confidence intervals (95%CIs) were calculated using a fixed- or random-effects model. Heterogeneity was represented by PH; publication bias and sensitivity analysis were also explored. Overall, no significant associations were found for any genetic models tested. However, upon stratification by cancer type, a significant decreased risk of breast cancer risk was identified in the homozygote comparison (OR = 0.79, 95%CI = 0.65-0.97 for TT versus CC). An analysis stratified by ethnicity and source of controls revealed an obvious decrease in risk among Asian groups in all genetic models, and among population-based controls only in the homozygote comparison and recessive model. Therefore, our meta-analysis suggested that the SHMT1 C1420T polymorphism was associated with decreased risk of breast cancer. Significant protective effects were found among Asian populations, but not in Caucasian groups. Due to some minor limitations, our findings should be confirmed by further studies.