[Changes and clinical significance of erythrocyte lifespan in megaloblastic anemia].

Objective: To investigate the lifespan of erythrocytes in megaloblastic anemia (MA) patients. Methods: A prospective cohort study analysis. Clinical data from 42 MA patients who were newly diagnosed at the Department of Hematology, Lanzhou University Second Hospital from January 2021 to August 2021 were analyzed, as were control data from 24 healthy volunteers acquired during the same period. The carbon monoxide breath test was used to measure erythrocyte lifespan, and correlations between erythrocyte lifespan and laboratory test indexes before and after treatment were calculated. Statistical analysis included the t-test and Pearson correlation. Results: The mean erythrocyte lifespan in the 42 newly diagnosed MA patients was (49.05±41.60) d, which was significantly shorter than that in the healthy control group [(104.13±42.62) d; t=5.13,P=0.001]. In a vitamin B12-deficient subset of MA patients the mean erythrocyte lifespan was (30.09±15.14) d, and in a folic acid-deficient subgroup it was (72.00±51.44) d, and the difference between these two MA subsets was significant (t=3.73, P=0.001). The mean erythrocyte lifespan after MA treatment was (101.28±33.02) d, which differed significantly from that before MA treatment (t=4.72, P=0.001). In MA patients erythrocyte lifespan was positively correlated with hemoglobin concentration (r=0.373), and negatively correlated with total bilirubin level (r=-0.425), indirect bilirubin level (r=-0.431), and lactate dehydrogenase level (r=-0.504) (all P<0.05). Conclusions: Erythrocyte lifespan was shortened in MA patients, and there was a significant difference between a vitamin B12-deficient group and a folic acid-deficient group. After treatment the erythrocyte lifespan can return to normal. Erythrocyte lifespan is expected to become an informative index for the diagnosis and treatment of MA.

Automatic three-dimensional segmentation of mouse embryonic stem cell nuclei by utilising multiple channels of confocal fluorescence images.

Time-lapse confocal fluorescence microscopy images from mouse embryonic stem cells (ESCs) carrying reporter genes, histone H2B-mCherry and Mvh-Venus, have been used to monitor dynamic changes in cellular/differentiation characteristics of live ESCs. Accurate cell nucleus segmentation is required to analyse the ESC dynamics and differentiation at a single cell resolution. Several methods used concavities on nucleus contours to segment overlapping cell nuclei. Our proposed method evaluates not only the concavities but also the size and shape of every 2D nucleus region to determine if any of the strait, extrusion, convexity and large diameter criteria is satisfied to segment overlapping nuclei inside the region. We then use a 3D segmentation method to reconstruct simple, convex, and reasonably sized 3D nuclei along the image stacking direction using the radius and centre of every segmented region in respective microscopy images. To avoid false concavities on nucleus boundaries, fluorescence images of the H2B-mCherry reporter are used for localisation of cell nuclei and Venus fluorescence images are used for determining the cell colony ranges. We use a series of image preprocessing procedures to remove noise outside and inside cell colonies, and in respective nuclei, and to smooth nucleus boundaries based on the colony ranges.  We propose dynamic data structures to record every segmented nucleus region and solid in sets (volumes) of 3D confocal images. The experimental results show that the proposed image preprocessing method preserves the areas of mouse ESC nuclei on microscopy images and that the segmentation method effectively segment out every nucleus with a reasonable size and shape. All 3D nuclei in a set (volume) of confocal microscopy images can be accessed by the dynamic data structures for 3D reconstruction. The 3D nuclei in time-lapse confocal microscopy images can be tracked to calculate cell movement and proliferation in consecutive volumes for understanding the dynamics of the differentiation characteristics about ESCs. LAY DESCRIPTION: Embryonic stem cells (ESCs) are considered as an ideal source for basic cell biology study and producing medically useful cells in vitro. This study uses time-lapse confocal fluorescence microscopy images from mouse ESCs carrying reporter gene to monitor dynamic changes in cellular/differentiation characteristics of live ESCs. To automate analyses of ESC differentiation behaviours, accurate cell nucleus segmentation to distinguish respective cells are required. A series of image preprocessing procedures are implemented to remove noise in live-cell fluorescence images but yield overlapping cell nuclei. A segmentation method that evaluates boundary concavities and the size and shape of every nucleus is then used to determine if any of the strait, extrusion, convexity, large and local minimum diameter criteria satisfied to segment overlapping nuclei. We propose a dynamic data structure to record every newly segmented nucleus. The experimental results show that the proposed image preprocessing method preserves the areas of mouse ESC nuclei and that the segmentation method effectively detects overlapping nuclei. All segmented nuclei in confocal images can be accessed using the dynamic data structures to be visualised and manipulated for quantitative analyses of the ESC differentiation behaviours. The manipulation can be tracking of segmented 3D cell nuclei in time-lapse images to calculate their dynamics of differentiation characteristics.

[Level and trend of respiratory diseases mortality in China from 2002 to 2016].

Objective: To analyze the level and trend of respiratory disease mortality in China from 2002 to 2016. Methods: The standardized mortality rates were calculated based on the China health statistics yearbook (2003-2012) and China statistical yearbook of health and family planning (2013-2017) data released by the statistical information center of National health Commission of the People's Republic of China. Joinpoint model was used to calculate the standardized mortality rates (SMR), Annual percentage change (APC) and the average annual percentage change (AAPC) for standardized mortality rates. Results: The SMR of respiratory diseases and chronic lower respiratory diseases were decreased significantly in 2002 to 2016 (AAPC=-3.6%, AAPC=-6.4%, P<0.001, respectively). The SMR of lung cancer showed a significant increase trend (AAPC=1.6%, P=0.001). There were no significant differences in the SMR of pneumonia and pneumonoconiosis (APCC=1.0%, P=0.242; APCC=-0.2%, P=0.905). Both urban and rural SMR of respiratory diseases were declining significantly (AAPC=-2.9%, P=0.001; AAPC=-4.2%, P<0.001). Both urban and rural SMR of lung cancer showed an increasing trend (AAPC=0.6%, P=0.022; AAPC=2.1%, P=0.003, respectively). The SMR of pneumonia in urban areas showed an upward trend (AAPC=2.7%, P=0.017). The SMR of respiratory disease of all age groups (<35 years old, 35-65 years old and ≥65 years old) showed a downward trend (AAPC=-3.8%, P=0.001; AAPC=-2.6%, P<0.001; AAPC=-3.9%, P<0.001). The SMR of pneumonia between 35 and 65 years old and SMR of lung cancer over 65 years old showed an increasing trend (AAPC=2.8%, P=0.001; AAPC=2.4%, P<0.001). The SMR of respiratory diseases among males and females showed a downtrend (AAPC=-3.1%, P<0.001; AAPC=-4.3%, P<0.001). However, the SMR of lung cancer in males and females increased significantly (AAPC=1.2%, P<0.001; AAPC=2.5%, P<0.001, respectively). There were no significant trends in the SMR of pneumonia and pneumoconiosis in males (AAPC=1.5%, P=0.096; AAPC=-1.6%, P=0.218). There was no obvious trend in the SMR of pneumonia in females (AAPC=-0.1%, P=0.872). Conclusions: The SMR of respiratory diseases in China generally shows a downward trend. The overall SMR and SMR of major respiratory diseases varies among different regions, genders and age groups.

[Prospective cohort study on the risks of pre-pregnancy overweight, excessive gestational weight gain on macrosomia].

Objective: To investigate the risks of pre-pregnancy overweight, excessive gestational weight gain on macrosomia. Methods: We conducted one hospital-based cohort study, focusing on pregnant women from January 2015. All pregnant women attending to this hospital for maternal check-ups, were included in our cohort and followed to the time of delivery. Data related to general demographic characteristics, pregnancy and health status of those pregnant women, was collected and maternal pre-pregnant BMI and maternal weight gain were calculated. Logistic regression was used to explore the risk difference of pre-pregnancy BMI, excessive gestational weight gain on macrosomia. Results: The overall incidence of macrosomia in our cohort appeared as 6.6% (149/2 243). After adjusting the confounding factors including age and histories on pregnancy, pre-pregnancy overweight/obesity was associated with higher risks of macrosomia (OR=3.12, 95%CI: 1.35-7.22, P=0.008; OR=2.99, 95%CI: 1.17-7.63, P=0.022) when comparing to those with normal pre-pregnancy weight. Cesarean delivery and sex of the offspring were associated with higher risk of macrosomia, while excessive gestational weight gain showed no significant difference (OR=1.41, 95%CI: 0.96-2.09, P=0.084). Our data showed that Macrosomia was statistically associated with gestational weight gain (P=0.002). After controlling parameters as age, history of pregnancy and related complications of the pregnant women, results from the logistic regression showed that women with gestational inadequate weight gain having reduced risks to deliver macrosomia, when compared to those pregnant women with adequate weight gain (OR=0.52, 95%CI: 0.30-0.90, P=0.019). Conclusion: Pre-pregnancy overweight and obesity were on higher risks to macrosomia.

[Downregulated PI3K-Akt-eNOS expression is related to increased atrial fibrillation inducibility in diabetic rats].

Objective: To explore the impact of PI3K-Akt-eNOS signaling on atrial fibrillation inducibility in diabetic rats. Method: Eight-week-old male diabetic rats were randomized assigned into GK group, IGF group and L-NAME group (n=8 each) which respectively received normal saline (NS), insulin like growth factor (IGF-1) or L-NAME+IGF-1 through tail vein daily for 4 weeks. Eight 8-week-old male homologous Wister-Kyoto(WKY) rats treated with intravenous NS served as control group (WKY group). Blood glucose was measured once per week. The left atrial diameter (LAD) was measured by echocardiography, the atrial electrical parameters, including the P-wave duration, the atrial effective refractory period (AERP) and its dispersion (AERP-d), the incidence and the duration of atrial fibrillation induced by atrial burst pacing, were evaluated by electro-physiological instrument at 4 weeks post various treatments. Rats were then sacrificed, left atrial (LA) cell morphology was determined on HE stained sections, LA interstitial collagen was determined on Masson stained sections. The protein expression of phosphatidylinositol 3-kinase (PI3K) and phosphate endothelial nitric oxide synthase (p-eNOS) were detected by Western blot. Results: (1) At the beginning of the study, the random blood glucose (GLU) level was significantly higher and LAD was large in GK, IGF and L-NAME groups than in WKY group;after 4 weeks, GLU level and LAD dimension of IGF group were lower than GK and L-NAME groups (P<0.01 or 0.05). (2) One rat in L-NAME group died during operation. Four weeks later, the incidence of atrial fibrillation in GK group, IGF group, L-NAME group and WKY group was 7/8, 2/8, 6/7 and 3/8. The median duration of atrial fibrillation in GK group, IGF group, L-NAME group and WKY group was 11.9(9.3, 13.1), 0(0, 1.8), 11.5(4.4, 12.0), and 0(0, 3.0) s. Compare with WKY group, the P-wave duration and PR interval were significantly longer, AERP-d, incidence, and duration of atrial fibrillation were significantly higher in GK group (P<0.01), these changed were significantly reversed in IGF group compared to GK and L-NAME groups (all P<0.01). Heart rate and AERP were similar among the 4 groups on (P>0.05). (3) Four weeks later, the CSA and CVF of LA were significantly larger in GK group than in WKY group (P<0.01), which were significantly reversed in IGF group (P<0.01 vs. GK group), and the beneficial effects of IGF disappeared by co-treatment with L-NAME (P<0.01 vs. IGF group). (4) Four weeks later, compare with WKY group, the protein expression of PI3K (P<0.01) and p-eNOS (P<0.05) of LA were significantly downregulated in GK group, which could be significantly upregulated by IGF (P<0.01 and 0.05 vs. GK group), these effects diminished by co-treatment with L-NAME (P<0.01 or 0.05 vs. IGF group). Conclusion: Increased atrial fibrillation susceptibility in diabetic rat is linked with structural and electrical remodeling in LA, possibly mediated through downregulated PI3K-Akt-eNOS signaling.

[Impact of the implementation of Beijing Tobacco Control Regulation on outpatient visits for smoking cessation in Beijing Chaoyang Hospital].

Objective: To investigate the impact of the implementation of Beijing Tobacco Control Regulation on outpatient visits for smoking cessation in Beijing Chaoyang Hospital. Methods: The outpatient visits and the proportion of returned cases to the total outpatient visits for smoking cessation in Beijing Chaoyang Hospital in past five years (2012-2016) that was before and after the implementation of Beijing Tobacco Control Regulation (June 1(st,) 2015) were analyzed. Time series seasonal index method was used to analyze seasonal variations in the outpatient visits. Results: From the implementation of Beijing Tobacco Control Regulation to December 31(th,) 2016, the average monthly outpatient visits for smoking cessation was significantly increased compared to that before the implementation [(101±37) vs (48±17), t=-7.486, P<0.001]. Meanwhile, the proportion of returned cases to the total outpatient visits was also significantly increased compared to that before the implementation [30.4% (582/1 912) vs 8.2% (161/1 966); χ(2)=309.8, P<0.001]. In addition, there were seasonal fluctuations in the number of outpatients from 2012 to 2016 , during which the outpatient visits started to increase in the second and third quarter (summer and fall) and peaked in June, but became less in the first and fourth quarter (spring and winter), and reached its nadir in January. Conclusions: With the implementation of Beijing Tobacco Control Regulation, the outpatient visits for smoking cessation have increased. Moreover, there is seasonal fluctuation in the outpatient visits for smoking cessation.

Reproductive biology of female bigeye tuna Thunnus obesus in the western Pacific Ocean.

The reproductive biology of female bigeye tuna Thunnus obesus was assessed by examining 888 fish (ranging from 84·9 to 174·4 cm fork length, LF ) caught by Taiwanese offshore longliners in the western Pacific Ocean from November 1997 to November 1998 and November to December 1999 and 258 gonad samples from these fish. The overall sex ratio of the catch during the sampling differed significantly from 0·5, but males were predominant in sizes >140 cm LF . Reproductive activity (assessed by histology), a gonado-somatic index, and the size-frequency distributions of whole oocytes indicated that spawning occurred throughout the year and the major spawning season appeared to be from February to September. The estimated sizes at 50% maturity (LF50 ) of females was 102·85 cm (95% c.i.: 90·79-110·21 cm) and the smallest mature female was 99·7 cm LF . They are multiple spawners and oocytes develop asynchronously. The proportion of mature (0·63) and reproductively active (0·70) females with ovaries containing postovulatory follicles indicated that they spawn almost daily. Batch fecundity for 15 females with the most advanced oocytes (>730 µm) ranged from 0·84 to 8·56 million eggs (mean ± s.d. = 3·06 ± 2·09). The relationships between batch fecundity (FB , in millions of eggs) and LF (cm) and round mass (MR , kg) were FB=9·91×10-14LF6·38 (r(2) = 0·84) and FB=8·89×10-4MR2·05 (r(2) = 0·80), respectively. The parameters estimated in this study are key information for stock assessments of T. obesus in the western Pacific Ocean and will contribute to the conservation and sustainable yield of this species.

Haemolytic disease of the newborn due to maternal irregular antibodies in the Chinese population in Taiwan.

From 1991 to 2000, amongst 23,886 full-term healthy Chinese babies delivered at our hospital, 2615 babies developed neonatal hyperbilirubinaemia. After excluding other causes of hyperbilirubinaemia and identifying the irregular antibodies, 15 cases of haemolytic disease of the newborn (HDN) due to maternal irregular antibodies were diagnosed; three cases were born in our hospital and 12 cases were referred. Amongst these 15 babies, six cases had HDN due to anti-E, three cases due to anti-E + c, three cases due to anti-D, one case due to anti-c and two cases due to 'Mi' antibodies reacting with MiIII phenotype cells (anti-Hil and anti-Mur). Although there were four cases of hydrops fetalis, only one of the patients expired. The prevalence of HDN caused by maternal irregular antibodies has been estimated to be 0.01%. Therefore, routine prenatal screening for irregular antibodies was not rational in the Chinese population in Taiwan. Anti-E and anti-E + c were the important irregular antibodies resulting in HDN. Although few cases of HDN due to anti-'Mi' have been reported, Anti-'Mi' is significant in regions with a high prevalence of the MiIII phenotype.

alpha-Adducin gene and essential hypertension in China.

Adducin is a membrane skeletal protein that is involved in the regulation of membrane ion transport and cellular signal transduction. Essential hypertension has been linked to alpha-adducin gene locus, and association of a polymorphism of the gene has been found in some studies, but results of linkage or association studies on alpha-adducin gene are controversial among different populations. This study was designed to examine the linkage between alpha-adducin gene locus and essential hypertension and to reveal the relationship between an alpha-adducin gene polymorphism (Gly460Trp) and essential hypertension in a Chinese population. For the linkage study, one hundred and six Chinese nuclear families were recruited, including 417 hypertensive patients in all 474 individuals. Those samples were genotyped at D4S412 and D4S3038. The distances between the two microsatellite markers and the alpha-adducin gene locus are less than 3cM. Parametric, non-parametric linkage (NPL) analyses using the GENEHUNTER software were carried out. Sib transmission-dise- quilibrium test (S-TDT), as well as transmission-disequilibrium test (TDT). was also implemented with TDT/S-TDT Program 1.1. Serum levels of uric acid, creatinine, blood urea nitrogen (BUN), fasting glucose and lipids were determined as phenotypes. In an association study, 138 hypertensive and 121 normotensive subjects were genotyped at Gly460Trp of the alpha-adducin gene to examine a possible association between this polymorphism and blood pressure or other phenotypes. We fail to find the linkage between the two markers and essential hypertension by parametric, NPL analysis or TDT/S-TDT study. With the use of the simple association and the multivariate logistic regression analyses, we also fail to reveal a significant association between the Gly460Trp polymorphism in alpha-adducin gene and the blood pressure variation, or blood biochemical indices studied. The frequency of the 460Trp allele in Chinese (46-48%) is similar to that found in Japanese (54-60%) while the allele frequency is less common in Caucasian (13%-23%). These findings suggest that in our Chinese population, alpha-adducin 460Trp variant may not play an important role in the etiology of EH. And the negative results of linkage and TDT/ S-TDT further supports this conclusion.

Linkage of hypertension to chromosome 2q14-q23 in Chinese families.

OBJECTIVES: To identify chromosome regions containing hypertension susceptibility genes in Chinese. SUBJECTS AND METHODS: A three-stage study was carried out in Chinese siblings ascertained through outpatient clinics. In the first stage, 283 affected sib-pairs from 79 nuclear families were subjected to a genome-wide scan with 240 microsatellite marker loci. The second stage focused on chromosome 2 with additional markers resulting in an average distance of 5 cM and used an independent sample of 637 affected sib-pairs from 161 families. In the third stage, a fine-scale mapping study on the suggestive region was performed in an independent set of 777 affected sib-pairs from 106 families. Fourteen markers were used with an average distance less than 2 cM. Non-parametric linkage analyses (NPL), parametric linkage analyses and transmission-disequilibrium tests were used to assess evidence for linkage and association. RESULTS: Three markers (D2S168 at 27.06 cM, D2S151 at 152.04 cM and D2S142 at 161.26 cM) on chromosome 2 with suggestive linkage to hypertension susceptibility genes were identified in the genome-wide scan. In stage II, the suggestive region around D2S151 and D2S142 was replicated, while the linkage around D2S168 was not. In the stage III fine-scale mapping study, multipoint linkage analyses showed LOD scores greater than 2.0 throughout a region between 157.16 cM and 162.46 cM (all P < 0.001) with a maximum peak of 2.24 (P= 0.00067) at 160.52 cM. We also observed a NPL Z-score peak of 3.27 at 157.55 cM (P= 0.00086). CONCLUSIONS: The results of a suggestive region on chromosome 2q14-q23 (D2S112-D2S2370) were consistent between each of the three studies. Interestingly, this region overlaps a syntenic region that contains blood pressure quantitative trait loci identified in rat models of hypertension. These data suggest that the region near D2S142 and D2S151 deserves to be further screened for hypertension susceptibility genes.