AIM: To characterize the peripheral T-cell subpopulation profiles and their correlation with hepatitis B virus (HBV) replication in different dinical stages of chronic HBV infection. METHODS: A total of 422 patien...AIM: To characterize the peripheral T-cell subpopulation profiles and their correlation with hepatitis B virus (HBV) replication in different dinical stages of chronic HBV infection. METHODS: A total of 422 patients with chronic HBV infection were enrolled in this study. The patients were divided into three stages: immune-tolerant stage, immune active stage, and immune-inactive carrier stage. Composition of peripheral T-cell subpopulations was determined by flow cytometry. HBV markers were detected by enzyme-linked immunosorbent assay. Serum HBV DNA load was assessed by quantitative real-time poiymerase chain reaction.RESULTS: CD8^+ T-cells were significantly higher in patients at the immune-tolerant stage than in patients at the immune-active and -inactive carrier stages (36.87 ± 7.58 vs 34.37 ± 9.07, 36.87 ± 7.58 vs 28.09 ± 5.64, P 〈 0.001). The peripheral blood in patients at the immune-tolerant and immune active stages contained more CD8^+ T-cells than CD4^+ T-cells (36.87 ± 7.58 vs 30.23 ± 6.35, 34.37 ± 9.07 vs 30.92 ± 7.40, P 〈 0.01), whereas the peripheral blood in patients at the immune- inactive carrier stage and in normal controls contained less CD8^+ T-cells than CD4^+ T-cells (28.09 ± 5.64 vs 36.85 ±6.06, 24.02 ± 4.35 vs 38.94 ± 3.39, P 〈 0.01). ANOVA linear trend test showed that CD8^+ T-cells were significantly increased in patients with a high viral load (39.41 ± 7.36, 33.83 ± 7.50, 31.81 ± 5.95 and 26.89 ± 5.71, P 〈 0.001), while CD4^+ T-cells were significantly increased in patients with a low HBV DNA load (37.45 ± 6.24, 33.33 ± 5.61, 31.58 ± 6.99 and 27.56 ± 5.49, P 〈 0.001). Nultiple regression analysis displayed that log copies of HBV DNA still maintained its highly significant coefficients for T-cell subpopulations, and was the strongest predictors for variations in CD3^+, CD4^+ and CD8^+ cells and CD4^+/CD8^+ ratio after adjustment for age at HBV-infection, maternal HBV-infection status, presence of hepatitis B e antigen and HBV mutation.CONCLUSION: Differences in peripheral T-cell subpopulation profiles can be found in different clinical stages of chronic HBV infection. T-cell impairment is significantly associated with HBV load.展开更多
AIM:To investigate the peripheral T-lymphocyte subpopulation profile,and its correlations with hepatitis B virus(HBV) replication level in chronic HBV-infected(CHI) individuals with normal liver function tests(LFTs) ....AIM:To investigate the peripheral T-lymphocyte subpopulation profile,and its correlations with hepatitis B virus(HBV) replication level in chronic HBV-infected(CHI) individuals with normal liver function tests(LFTs) . METHODS:Frequencies of T-lymphocyte subpopu-lations in peripheral blood were measured by flow cytometry in 216 CHI individuals. HBV markers were detected with ELISA. Serum HBV DNA load was assessed with quantitative real-time PCR. Information of age at HBV infection,and maternal HBV infection status was collected. ANOVA linear trend test and linear regression were used in statistical analysis. RESULTS:CHI individuals had significantly decreased relative frequencies of CD3+,CD4+ subpopulationsand CD4+/CD8+ ratio,and increased CD8+ subset percentage compared with uninfected individuals(all P < 0.001) . There was a significant linear relationship between the load of HBV DNA and the parameters of T-lymphocyte subpopulations(ANOVA linear trend test P < 0.01) . The parameters were also significantly worse among individuals whose mothers were known to be HBV carriers,and those having gained infection before the age of 8 years. In multiple regressions,after adjustment for age at HBV infection and status of maternal HBV infection,log copies of HBV DNA maintained its highly significant predictive coefficient on T-lymphocyte subpopulations,whereas the effect of HBeAg was not significant. CONCLUSION:HBV DNA correlates with modification in the relative T-lymphocyte subpopulation frequencies. High viral load is more powerful than HBeAg in predicting the impaired balance of T-cell subsets.展开更多
Objective: To investigate the potential and early effect of hypertonic saline resuscitation on T-lymphocyte sub- populations in rats with hemorrhagic shock. Methods: A model of rat with severe hemorrhagic shock was es...Objective: To investigate the potential and early effect of hypertonic saline resuscitation on T-lymphocyte sub- populations in rats with hemorrhagic shock. Methods: A model of rat with severe hemorrhagic shock was established in 18 Sprague-Dawley (SD) rats. The rats were randomly divided into Sham group, HTS group (hypertonic saline resuscitation group) and NS group (normal saline resuscitation group). Each group contained 6 rats. The CD4+ and CD8+ subpopulations of T-lymphocytes in peripheral blood were detected respectively before shock and after resuscitation by double antibody labelling and flow cytometry. Results: In the early stage after hemorrhagic shock, fluid resuscitation and emergency treatment, the CD4+ lymphocytes of peripheral blood in HTS and NS groups markedly increased. Small volume resuscitation with HTS also induced peripheral CD8+ lymphocytes to a certain extent, whereas NS resuscitation showed no effect in this respect. Consequently, compared with Sham and HTS groups, CD4+/CD8+ ratio of peripheral blood in NS group was obviously increased, and showed statistically differences. Conclusion: In this model of rat with severe hemorrhagic shock, small volume resuscitation with HTS is more effective than NS in reducing immunologic disorders and promoting a more balanced profile of T-lymphocyte subpopula- tions regulating network.展开更多
Pig-tailed macaques(Macaca nemistrina group) have been extensively used as non-human primate animal models for various human diseases in recent years, notably for AIDS research due to their sensitivity to HIV-1. North...Pig-tailed macaques(Macaca nemistrina group) have been extensively used as non-human primate animal models for various human diseases in recent years, notably for AIDS research due to their sensitivity to HIV-1. Northern pig-tailed macaques(M. leonina) are distributed in China and other surrounding Southeast Asia countries. Although northern pig-tailed macaques have been bred on a large scale as experimental animals since 2012, the reference value of normal levels of leukocytes is not available. To obtain such information, 62 blood samples from male and female healthy northern pig-tailed macaques at different ages were collected. The normal range of major leukocyte subpopulations, such as T lymphocytes, B lymphocytes, natural killer(NK) cells, monocytes, and the expression levels of activation or differentiation related molecules(CD38, HLA-DR, CCR5, CD21, IgD, CD80 and CD86) on lymphocytes were analyzed by flow cytometry. The counts of B cells decreased with age, but those of CD8+ T cells and NK cells and the frequency of CD38+HLA-DR+CD4+ T cells were positively correlated with age. The counts of leukocyte subpopulations were higher in males than those in females except for CD4+ T cells. Males also showed higher expression levels of Ig D and CD21 within B cells. This study provides basic data about the leukocyte subpopulations of northern pig-tailed macaques and compares this species with commonly used Chinese rhesus macaques(M. mulatta), which is meaningful for the biomedical application of northern pig-tailed macaques.展开更多
Hand osteoarthritis is a common heterogeneous joint disorder with unclear molecular mechanisms and no disease-modifying drugs.In this study,we performed single-cell RNA sequencing analysis to compare the cellular comp...Hand osteoarthritis is a common heterogeneous joint disorder with unclear molecular mechanisms and no disease-modifying drugs.In this study,we performed single-cell RNA sequencing analysis to compare the cellular composition and subpopulationspecific gene expression between cartilage with macroscopically confirmed osteoarthritis(n=5)and cartilage without osteoarthritis(n=5)from the interphalangeal joints of five donors.Of 105142 cells,we identified 13 subpopulations,including a novel subpopulation with inflammation-modulating potential annotated as inflammatory chondrocytes.Fibrocartilage chondrocytes exhibited extensive alteration of gene expression patterns in osteoarthritic cartilage compared with nonosteoarthritic cartilage.Both inflammatory chondrocytes and fibrocartilage chondrocytes showed a trend toward increased numbers in osteoarthritic cartilage.In these two subpopulations from osteoarthritic cartilage,the ferroptosis pathway was enriched,and expression of iron overload-related genes,e.g.,FTH1,was elevated.To verify these findings,we conducted a Mendelian randomization study using UK Biobank and a population-based cross-sectional study using data collected from Xiangya Osteoarthritis Study.Genetic predisposition toward higher expression of FTH1 mRNA significantly increased the risk of hand osteoarthritis(odds ratio=1.07,95%confidence interval:1.02–1.11)among participants(n=332668)in UK Biobank.High levels of serum ferritin(encoded by FTH1),a biomarker of body iron overload,were significantly associated with a high prevalence of hand osteoarthritis among participants(n=1241)of Xiangya Osteoarthritis Study(P-for-trend=0.037).In conclusion,our findings indicate that inflammatory and fibrocartilage chondrocytes are key subpopulations and that ferroptosis may be a key pathway in hand osteoarthritis,providing new insights into the pathophysiology and potential therapeutic targets of hand osteoarthritis.展开更多
2,4-dinitrophenol (DNP), an organic compound which frequently used in industry, is considered to have high toxicity. This study aimed to investigate the early changes of lymphocyte subpopulations in patients with oc...2,4-dinitrophenol (DNP), an organic compound which frequently used in industry, is considered to have high toxicity. This study aimed to investigate the early changes of lymphocyte subpopulations in patients with occupational 2,4-DNP poisoning. Totally 9 patients with acute occupational 2,4-DNP poisoning and 30 healthy volunteers as control were enrolled. The patients received immediately comprehensive supportive treatments, including large-dose glucocorticoid and repeated hemoperfusion (HP). The ratio of CD4+/CD8+ T cells were significantly higher in patients upon admission compared to healthy controls (P 〈 0.01); however, counts of total lymphocytes, CD3+, CD3+CD4+, CD3+CD8+, B (CD19+), and natural killer (NK) cells (CD16+CD56+) were significantly reduced (all P 〈 0.001). The NK cell count was negatively correlated with initial plasma 2,4-DNP concentration (r = -0.750, P = 0.026). Thus, acute occupational 2,4-DNP poisoning was accompanied by immediate complex immune cell reactions, especially NK cells might play important role in severe 2,4-DNP poisoning.展开更多
This study was designed to analyze the sperm kinematic and morphometric subpopulations in the different fractions of the ejaculate in normozoospermic men. Ejaculates from eight normozoospermic men were collected by ma...This study was designed to analyze the sperm kinematic and morphometric subpopulations in the different fractions of the ejaculate in normozoospermic men. Ejaculates from eight normozoospermic men were collected by masturbation in three fractions after 3-5 days of sexual abstinence. Analyses of sperm motility by computer-assisted sperm analysis (CASA-Mot), and of sperm morphometry by computer-assisted sperm morphometry analysis (CASA-Morph) using fluorescence were performed. Clustering and discriminant procedures were performed to identify sperm subpopulations in the kinematic and morphometric data obtained. Clustering procedures resulted in the classification of spermatozoa into three kinematic subpopulations (slow with low ALH [35.6% of all motile spermatozoa], with circular trajectories [32.0%], and rapid with high ALH [32.4%]), and three morphometric subpopulations (large-round [33.9% of all spermatozoa], elongated [32.0%], and small [34.10%]). The distribution of kinematic sperm subpopulations was different among ejaculate fractions (P 〈 0.001), with higher percentages of spermatozoa exhibiting slow movements with low ALH in the second and third portions, and with a more homogeneous distribution of kinematic sperm subpopulations in the first portion. The distribution of morphometric sperm subpopulations was also different among ejaculate fractions (P〈 0.001), with more elongated spermatozoa in the first, and of small spermatozoa in the third, portion. It is concluded that important variations in the distribution of kinematic and morphometric sperm subpopulations exist between ejaculate fractions, with possible functional implications.展开更多
OECs for spinal cord repair: Is repairing the iniured spinal cord by olfactory ensheathing cell (OEC) transplantation pos- sible? A recent human trial in which a paralysed man regained some function after transpla...OECs for spinal cord repair: Is repairing the iniured spinal cord by olfactory ensheathing cell (OEC) transplantation pos- sible? A recent human trial in which a paralysed man regained some function after transplantation of partially purified OECs suggests that this therapy may be a successful approach (Ta- bakow et al., 2014). In another human trial in which olfactory mucosa lamina propria was transplanted, patients recovered some motor and sensory function (Wang et al., 2015). While these results show promise, it is clear that improvements are needed to provide patients with increased functional output. Strategies to improve the therapeutic use of OECs may include improving the purification of the OECs used for transplantation, using them in combination with growth factors to combat the inhibitory environment and improve anon growth, the use of nerve bridges, advanced physiotherapy and the use of exo- skeleton robotics to reinforce functional connections. Of all these approaches, it is probably is primarily addressed to ensure crucial that the purity of OECs consistency in outcomes.展开更多
Teratozoospermia (〈40% morphologically normal spermatozoa/ejaculate) is a frequent phenomenon in feline species. This research was carried out to study the possible differences in testicular volume, differential sp...Teratozoospermia (〈40% morphologically normal spermatozoa/ejaculate) is a frequent phenomenon in feline species. This research was carried out to study the possible differences in testicular volume, differential sperm morphometric traits, and potential differences regarding the sperm subpopulational structure during epididymal sperm maturation in teratozoospermic feline donors. Epididymal sperm samples were collected from the caput (R1), corpus (R2), and cauda (R3) epididymidis in two donor groups (N: normozoospermic; T: teratozoospermic). Aliquots were assessed for concentration, viability, motility, and acrosomal integrity. Sperm morphometric descriptors from CASA-Morph analysis were analyzed by the Principal Component Analysis (PCA) and clustering analyses. Irrespective of the group analyzed, PCA revealed two Principal Components (PCs) for each epididymal region explaining more than the 93% of the variance. Surprisingly, the number of subpopulations remained constant in regions R1-R2-R3 irrespective of the donor group analyzed. However, the distribution of these subpopulations was found to be structurally different and strongly influenced by the epididymal region and the donor group. In conclusion, testicular morphometry and the sperm subpopulation structure were different in N and T donors. The alterations in subpopulations during epididymal maturation could be used as a potential clinical indicator of teratozoospermic individuals since an important influence of teratozoospermia on sperm subpopulation structure has been demonstrated.展开更多
AIMTo identify and characterize functionally distinct subpopulation of adipose-derived stem cells (ADSCs). METHODSADSCs cultured from mouse subcutaneous adipose tissue were sorted fluorescence-activated cell sorter ba...AIMTo identify and characterize functionally distinct subpopulation of adipose-derived stem cells (ADSCs). METHODSADSCs cultured from mouse subcutaneous adipose tissue were sorted fluorescence-activated cell sorter based on aldehyde dehydrogenase (ALDH) activity, a widely used stem cell marker. Differentiation potentials were analyzed by utilizing immunocytofluorescece and its quantitative analysis. RESULTSApproximately 15% of bulk ADSCs showed high ALDH activity in flow cytometric analysis. Although significant difference was not seen in proliferation capacity, the adipogenic and osteogenic differentiation capacity was higher in ALDH<sup>Hi</sup> subpopulations than in ALDH<sup>Lo</sup>. Gene set enrichment analysis revealed that ribosome-related gene sets were enriched in the ALDH<sup>Hi</sup> subpopulation. CONCLUSIONHigh ALDH activity is a useful marker for identifying functionally different subpopulations in murine ADSCs. Additionally, we suggested the importance of ribosome for differentiation of ADSCs by gene set enrichment analysis.展开更多
Sperm quality is evaluated for the calculation of sperm dosage in artificial reproductive programs. The most common parameter used is motility, but morphology has a higher potential as a predictor of genetic quality. ...Sperm quality is evaluated for the calculation of sperm dosage in artificial reproductive programs. The most common parameter used is motility, but morphology has a higher potential as a predictor of genetic quality. Morphometry calculations from CASA-Morph technology improve morphological evaluation and allow mathematical approaches to the problem. Semen from 28 Holstein bulls was collected by artificial vagina, and several ejaculates were studied. After general evaluation, samples were diluted, packaged in 0.25 ml straws, and stored in liquid nitrogen. Two straws per sample were thawed, and slides were processed and stained with Diff-Quik. Samples were analyzed by a CASA-Morph system for eight morphometric parameters. In addition to the "classical" statistical approach, based on variance analysis (revealing differences between animals, ejaculates, and straws), principal component (PC) analysis showed that the variables were grouped into PC1, related to size, and PC2 to shape. Subpopulation structure analysis showed four groups, namely, big, small, short, and narrow from their dominant characteristics, representing 31.0%, 27.3%, 24.1%, and 17.7% of the total population, respectively. The distributions varied between animals and ejaculates, but between straws, there were no differences in only four animals. This modern approach of considering an ejaculate sperm population as divided into subpopulations reflecting quantifiable parameters generated by CASA-Morph systems technology opens a new view on sperm function. This is the first study applying this approach to evaluate different ejaculates and straws from the same individual. More work must be done to improve seminal dose calculations in assisted reproductive programs.展开更多
This study was designed to characterize morphometric sperm subpopulations in normozoospermic men by using different statistical methods and examining their suitability to classify correctly different sperm nuclear mor...This study was designed to characterize morphometric sperm subpopulations in normozoospermic men by using different statistical methods and examining their suitability to classify correctly different sperm nuclear morphologies present in human ejaculates. Ejaculates from 21 normozoospermic men were collected for the study. After semen collection and analysis, samples were prepared for morphometric determination. At least 200 spermatozoa per sample were assessed for sperm morphometry by computer-assisted sperm morphometry analysis (CASA-Morph) using fluorescence. Clustering and discriminant procedures were performed to identify sperm subpopulations from the morphometric data obtained. Clustering procedures resulted in the classification of spermatozoa into three morphometric subpopulations (large-round 30.4%, small-round 46.6%, and large-elongated 22.9%). In the second analysis, using discriminant methods, the classification was made independently of size and shape. Three morphological categories according to nuclear size (small 〈10.90 μm^2, intermediate 10.91-13.07 μm^2, and large 〉13.07 μm^2) and four categories were defined on 400 canonical cells (100 × 4) from 10 men according to sperm nuclear shape (oval, pyriform, round, and elongated). Thereafter, the resulting classification functions were used to categorize 4200 spermatozoa from 21 men. Differences in the class distribution were observed among men from both clustering and discriminant procedures. It was concluded that the combination of CASA-Morph fluorescence-based technology with multivariate cluster or discriminant analyses provides new information on the description of different morphometric sperm subpopulations in normal individuals, and that important variations in the distribution of morphometric sperm subpopulations may exist between men, with possible functional implications.展开更多
A study of the genetic diversity within four subpopulations of Chimonobambusa rivularis Yi and adaptations to altitude, habitat and related factors in Qionglai City of Sichuan province in China, analyzed by random amp...A study of the genetic diversity within four subpopulations of Chimonobambusa rivularis Yi and adaptations to altitude, habitat and related factors in Qionglai City of Sichuan province in China, analyzed by random amplified polymorphic DNA technique. 22 random primers were selected in the amplification and 375 repetitive loci with 350 polymorphic loci were produced. The total average percentage of amplification loci was 93.3%. The genetic diversity of every subpopulation was medium on (the percentage of amplification loci was 49.33%-66.67%). The genetic diversity of high altitude samples of Zhengtiantai was lower than that of low altitude Wutonggang's. There was no obvious differentiation between two subpopulations of Zhengtiantais. The correlation is very low between altitude and the amplification loci and the genetic distance, Pearson correlation coefficient was 0.431 and 0.488 (P 〈 0.01). Through investigation, the primary cause of heredity multiplicity drops was that the habitat had been destroyed by tourism development and other human disturbance.展开更多
This work provides information on the blue fox ejaculated sperm quality needed for seminal dose calculations. Twenty semen samples, obtained by masturbation, were analyzed for kinematic and morphometric parameters by ...This work provides information on the blue fox ejaculated sperm quality needed for seminal dose calculations. Twenty semen samples, obtained by masturbation, were analyzed for kinematic and morphometric parameters by using CASA-Mot and CASA-Morph system and principal component (PC) analysis. For motility, eight kinematic parameters were evaluated, which were reduced to PC1, related to linear variables, and PC2, related to oscillatory movement. The whole population was divided into three independent subpopulations: SP1, fast cells with linear movement; SP2, slow cells and nonoscillatory motility; and SP3, medium speed cells and oscillatory movement. In almost all cases, the subpopulation distribution by animal was significantly different. Head morphology analysis generated four size and four shape parameters, which were reduced to PC 1, related to size, and PC2, related to shape of the cells. Three morphometric subpopulations existed. SPI: large oval cells; SP2: medium size elongated cells; and SP3. small and short cells. The subpopulation distribution differed between animals. Combining the kinematic and morphometric datasets produced PC1, related to morphometric parameters, and PC2, related to kinematics, which generated four sperm subpopulations - SPI.. high oscillatory motility, large and short heads; SP2; medium velocity with small and short heads; SP3. slow motion small and elongated cells; and SP4. high linear speed and large elongated cells. Subpopulation distribution was different in all animals. The establishment of sperm subpopulations from kinematic, morphometric, and combined variables not only improves the well-defined fox semen characteristics and offers a good conceptual basis for fertility and sperm preservation techniques in this species, but also opens the door to use this approach in other species, included humans.展开更多
Attribute reduction is an important process in rough set theory.Finding minimum attribute reduction has been proven to help the user-oriented make better knowledge discovery in some cases.In this paper,an efficient mi...Attribute reduction is an important process in rough set theory.Finding minimum attribute reduction has been proven to help the user-oriented make better knowledge discovery in some cases.In this paper,an efficient minimum attribute reduction algorithm is proposed based on the multilevel evolutionary tree with self-adaptive subpopulations.A model of multilevel evolutionary tree with self-adaptive subpopulations is constructed,and interacting attribute sets are better decomposed into subsets by the self-adaptive mechanism of elitist populations.Moreover it can self-adapt the subpopulation sizes according to the historical performance record so that interacting attribute decision variables are captured into the same grouped subpopulation,which will be extended to better performance in both quality of solution and competitive computation complexity for minimum attribute reduction.The conducted experiments show the proposed algorithm is better on both efficiency and accuracy of minimum attribute reduction than some representative algorithms.Finally the proposed algorithm is applied to magnetic resonance image(MRI)segmentation,and its stronger applicability is further demonstrated by the effective and robust segmentation results.展开更多
The main aims of this research were to study possible differences in objective morphometric sperm characteristics, establish normative sperm morphometry standards, and evaluate the presumed different subpopulation dis...The main aims of this research were to study possible differences in objective morphometric sperm characteristics, establish normative sperm morphometry standards, and evaluate the presumed different subpopulation distribution of avian spermatozoa from the rooster (Gallus domesticus) and Guinea fowl (Numida meleagris) as model avian species. Seventy-two ejaculates (36 per species studied) were obtained manually, following a training period involving gently combined dorso-abdominal and lumbo.sacral massage of the birds. Ejaculates were processed for volume, sperm concentration, viability, motility, and morphology. Moreover, samples were submitted for sperm morphometric assessment using objective Computer-Assisted Semen Analysis for Morphometry (CASA-Morph) methods, with sperm morphometric descriptors evaluated by Principal Component Analysis (PCA) and multivariate clustering analyses. There were several differences observed between the avian species in values obtained for ejaculate volume and sperm concentration (P 〈 0.001). Irrespective of species, PCA revealed two Principal Components (PCs) explaining more than 80% of the variance. In addition, the number of subpopulations differed with species (three and five subpopulations for rooster and Guinea fowl, respectively). Moreover, the distribution of the sperm subpopulations was found to be structurally different between species. In conclusion, our findings from using CASA-Morph methods indicate pronounced sperm morphometric variation between these two avian species. Because of the strong differences observed in morphometric parameter values and their subpopulation distribution, these results suggest that application of objective analytical methods such as CASA-Morph could substantially improve the reliability of comparative studies and help establish valid normative sperm morphological values for avian species.展开更多
This study aimed to identify and characterize the sperm subpopulations existing in water buffalo semen using a computer assisted sperm analyzer (CASA), as well as assess the effects of cryopreservation on the sperm ...This study aimed to identify and characterize the sperm subpopulations existing in water buffalo semen using a computer assisted sperm analyzer (CASA), as well as assess the effects of cryopreservation on the sperm subpopulation structure and evaluate bull variability. The semen of eight Bulgarian Murrah bulls was collected by four times in an interval of one week each. The semen was cryopreserved following a standard protocol and sperm kinematics was assessed. Clustering methods were applied to individual sperms, forming two significantly different (P 〈 0.05) subpopulations (P1 and P2). Subpopulation P1 represents those spermatozoa that moved most rapidly and progressively (46.29%), and subpopulation P2 includes spermatozoa with relatively low velocity or poorly motile but with high progressiveness (53.41%). There was a decline on the population of P1 sperms from fresh (52.52%), pre-freeze (45.73%) to post-thaw (35.17%) stages and significant difference on the sperm kinematics between P1 and P2. A significant decline in the values of distance, velocity and amplitude of lateral head (ALH) parameters were observed at post-thaw stage, while an increase was observed on trajectory and beat cross frequency (BCF) kinematics. Values of sperm kinematics were also significantly different (P 〈 0.05) among all bulls. The frequency distribution of spermatozoa on both subpopulations P1 and P2 was quite similar for all bulls in pre-freeze and post-thaw stages, but with significant (P 〈 0.05) variability on fresh stage. Bulls with the highest maintained frequency of P1 sperms are denoted as good freezer bulls. In sum, kinematic characterization of water buffalo sperm and clustering into subpopulation enable to identify bulls that are more resistant to cryopreservation and production of quality semen for genetic propagation.展开更多
Objective: To study the cellular immunity function of patientswith early syphilis and the effects on immune modifiersEsberitox N or IFN. Methods: T-lymphocyte subpopulations of the peripheralblood in 44 patients with ...Objective: To study the cellular immunity function of patientswith early syphilis and the effects on immune modifiersEsberitox N or IFN. Methods: T-lymphocyte subpopulations of the peripheralblood in 44 patients with syphilis and 40 healthy controls wereexamined by flow cytometry. Results: The number of CD_4+ cells and the CD_4+/CD_8+ ratioin patients with syphilis were found to be significantly lowerthan those in the control (P<0.0l), while the number of CD_8+cells was higher than that in the control (P<0.01). TheCD_4+/CD_8+ ratio in those with active disease was lower thanthat in those who had been cured (P<0.05). The CD_4+ countand the CD_4+/CD_8+ ratio in those treated with antibiotics alone(Penicillin G or Cephalosporins) were lower than those treatedwith both antibiotics and immunomodulators (P<0.05). Conclusions: Cellular immunity in the patients with earlysyphilis was prominently suppressed, and treatment withimmunomodulators may be helpful for the recovery of cellularimmunity of these patients.展开更多
<span style="font-family:Verdana;"><strong>Rationale</strong></span><b><span style="font-family:Verdana;">:</span></b><span style="font-famil...<span style="font-family:Verdana;"><strong>Rationale</strong></span><b><span style="font-family:Verdana;">:</span></b><span style="font-family:Verdana;"> The known prognosis factors for rhabdomyosarcoma (RMS) in</span><span style="font-family:;" "=""> <span style="font-family:Verdana;">children do not always explain the unsatisfactory outcome of treatment.</span><span style="font-family:Verdana;"> Changes in the subpopulation composition of Bone Marrow (BM) effector cells during </span><span style="font-family:Verdana;">the development of RMS may indicate new directions for the search for </span><span style="font-family:Verdana;">prog</span><span><span style="font-family:Verdana;">nostic factors and points for the impact of targeted therapy. </span><b><span style="font-family:Verdana;">Purpose</span></b></span></span><b><span style="font-family:Verdana;">:</span></b><span style="font-family:Verdana;"> To</span><span style="font-family:;" "=""><span style="font-family:Verdana;"> identify correlations between quantitative changes in the levels of subpopulations of T, B and NK-lymphocytes of BM and known risk factors for RMS in child</span><span><span style="font-family:Verdana;">ren. </span><b><span style="font-family:Verdana;">Objects</span></b></span></span><b><span style="font-family:Verdana;">:</span></b><span style="font-family:Verdana;"> The study included 31 patients. The main group included 16</span><span style="font-family:;" "=""><span style="font-family:Verdana;"> patients with RMS, average age—6.8 ± 1.0 years, while children 1 - 10 years old—13 (81.3%), over 10 years old—3 (18.8%) people, girls and boys were 8 </span><span style="font-family:Verdana;">people each. The embryonic variant of RMS was established in 10 (62.5%) </span><span style="font-family:Verdana;">cases, the alveolar variant—in 4 (25%) cases, in two patients (12</span></span><span style="font-family:Verdana;">.</span><span style="font-family:;" "=""><span style="font-family:Verdana;">5%), the histological variant was not established. In 12 (75%) patients, an unfavorable loca</span><span style="font-family:Verdana;">lization of the RMS (parameningeal, extremities, prostate, bladder) was re</span><span style="font-family:Verdana;">vealed, in 4 patients (25%), the localization of </span></span><span style="font-family:Verdana;">the </span><span style="font-family:;" "=""><span style="font-family:Verdana;">tumor was regarded as favorable. Patients with T2b—13 (81.2%) and T2a—2 (12.5%) stages prevailed. Regional and distant metastases were detected in 10 (52.6%) patients. The comparison group included 15 children in whom the presence of malignant neoplasia was excluded, the average age was 8.4 ± 1.5 years, 11 boys (73.7%) and 4 girls </span><span><span style="font-family:Verdana;">(26.3%). </span><b><span style="font-family:Verdana;">Methods</span></b></span></span><b><span style="font-family:Verdana;">:</span></b><span style="font-family:Verdana;"> All patients underwent morphological (myelogram</span><span style="font-family:;" "=""><span style="font-family:Verdana;"> count</span><span style="font-family:Verdana;">ing) and immunological (quantitative analysis of lymphocytic subpopula</span><span style="font-family:Verdana;">tions) bone marrow studies. Immunophenotyping in all patients was carried out by direct immunofluorescence using a triple fluorescent label. </span><b><span style="font-family:Verdana;">Results</span></b></span><b><span style="font-family:Verdana;">:</span></b><b><span style="font-family:;" "=""> </span></b><span style="font-family:;" "=""><span style="font-family:Verdana;">Significant differences in the levels of subpopulations of BM T-lymphocytes were found when comparing the values of the main group, distributed by localization and histological variant, with the data obtained in the control group of patients. For example, the percentage of </span><span style="font-family:Verdana;">CD3+ T cells with the co-stimulatory molecule CD28+ was significantly higher in patients with parameningeal </span><span style="font-family:Verdana;">RMS </span><span><span style="font-family:Verdana;">(p = 0.010). </span><b><span style="font-family:Verdana;">Conclusion</span></b></span></span><b><span style="font-family:Verdana;">:</span></b><span style="font-family:Verdana;"> Each clinical group of patients has its own individual immunological characteristics. The results obtained by us can be considered indicative and regarded as starting points for further study of the peculiarities of the subpopulation composition of BM in patients with RMS.</span>展开更多
文摘AIM: To characterize the peripheral T-cell subpopulation profiles and their correlation with hepatitis B virus (HBV) replication in different dinical stages of chronic HBV infection. METHODS: A total of 422 patients with chronic HBV infection were enrolled in this study. The patients were divided into three stages: immune-tolerant stage, immune active stage, and immune-inactive carrier stage. Composition of peripheral T-cell subpopulations was determined by flow cytometry. HBV markers were detected by enzyme-linked immunosorbent assay. Serum HBV DNA load was assessed by quantitative real-time poiymerase chain reaction.RESULTS: CD8^+ T-cells were significantly higher in patients at the immune-tolerant stage than in patients at the immune-active and -inactive carrier stages (36.87 ± 7.58 vs 34.37 ± 9.07, 36.87 ± 7.58 vs 28.09 ± 5.64, P 〈 0.001). The peripheral blood in patients at the immune-tolerant and immune active stages contained more CD8^+ T-cells than CD4^+ T-cells (36.87 ± 7.58 vs 30.23 ± 6.35, 34.37 ± 9.07 vs 30.92 ± 7.40, P 〈 0.01), whereas the peripheral blood in patients at the immune- inactive carrier stage and in normal controls contained less CD8^+ T-cells than CD4^+ T-cells (28.09 ± 5.64 vs 36.85 ±6.06, 24.02 ± 4.35 vs 38.94 ± 3.39, P 〈 0.01). ANOVA linear trend test showed that CD8^+ T-cells were significantly increased in patients with a high viral load (39.41 ± 7.36, 33.83 ± 7.50, 31.81 ± 5.95 and 26.89 ± 5.71, P 〈 0.001), while CD4^+ T-cells were significantly increased in patients with a low HBV DNA load (37.45 ± 6.24, 33.33 ± 5.61, 31.58 ± 6.99 and 27.56 ± 5.49, P 〈 0.001). Nultiple regression analysis displayed that log copies of HBV DNA still maintained its highly significant coefficients for T-cell subpopulations, and was the strongest predictors for variations in CD3^+, CD4^+ and CD8^+ cells and CD4^+/CD8^+ ratio after adjustment for age at HBV-infection, maternal HBV-infection status, presence of hepatitis B e antigen and HBV mutation.CONCLUSION: Differences in peripheral T-cell subpopulation profiles can be found in different clinical stages of chronic HBV infection. T-cell impairment is significantly associated with HBV load.
文摘AIM:To investigate the peripheral T-lymphocyte subpopulation profile,and its correlations with hepatitis B virus(HBV) replication level in chronic HBV-infected(CHI) individuals with normal liver function tests(LFTs) . METHODS:Frequencies of T-lymphocyte subpopu-lations in peripheral blood were measured by flow cytometry in 216 CHI individuals. HBV markers were detected with ELISA. Serum HBV DNA load was assessed with quantitative real-time PCR. Information of age at HBV infection,and maternal HBV infection status was collected. ANOVA linear trend test and linear regression were used in statistical analysis. RESULTS:CHI individuals had significantly decreased relative frequencies of CD3+,CD4+ subpopulationsand CD4+/CD8+ ratio,and increased CD8+ subset percentage compared with uninfected individuals(all P < 0.001) . There was a significant linear relationship between the load of HBV DNA and the parameters of T-lymphocyte subpopulations(ANOVA linear trend test P < 0.01) . The parameters were also significantly worse among individuals whose mothers were known to be HBV carriers,and those having gained infection before the age of 8 years. In multiple regressions,after adjustment for age at HBV infection and status of maternal HBV infection,log copies of HBV DNA maintained its highly significant predictive coefficient on T-lymphocyte subpopulations,whereas the effect of HBeAg was not significant. CONCLUSION:HBV DNA correlates with modification in the relative T-lymphocyte subpopulation frequencies. High viral load is more powerful than HBeAg in predicting the impaired balance of T-cell subsets.
文摘Objective: To investigate the potential and early effect of hypertonic saline resuscitation on T-lymphocyte sub- populations in rats with hemorrhagic shock. Methods: A model of rat with severe hemorrhagic shock was established in 18 Sprague-Dawley (SD) rats. The rats were randomly divided into Sham group, HTS group (hypertonic saline resuscitation group) and NS group (normal saline resuscitation group). Each group contained 6 rats. The CD4+ and CD8+ subpopulations of T-lymphocytes in peripheral blood were detected respectively before shock and after resuscitation by double antibody labelling and flow cytometry. Results: In the early stage after hemorrhagic shock, fluid resuscitation and emergency treatment, the CD4+ lymphocytes of peripheral blood in HTS and NS groups markedly increased. Small volume resuscitation with HTS also induced peripheral CD8+ lymphocytes to a certain extent, whereas NS resuscitation showed no effect in this respect. Consequently, compared with Sham and HTS groups, CD4+/CD8+ ratio of peripheral blood in NS group was obviously increased, and showed statistically differences. Conclusion: In this model of rat with severe hemorrhagic shock, small volume resuscitation with HTS is more effective than NS in reducing immunologic disorders and promoting a more balanced profile of T-lymphocyte subpopula- tions regulating network.
基金This work was supported by the National Special Science Research Program of China (2012CBA01305) the National Natural Science Foundation of China (81172876, U0832601, 81273251, U1202228) the Knowledge Innovation Program of CAS (KSCX2-EW-R-13) and the National Science and Technology Major Project (2013ZX10001-002, 2012ZX10001-007)We thank Mr. Zhen-Fei Hu,Gui Li and Dong-Ti Huang of Kunming Primate Research Center for their assistance with the experiments.
文摘Pig-tailed macaques(Macaca nemistrina group) have been extensively used as non-human primate animal models for various human diseases in recent years, notably for AIDS research due to their sensitivity to HIV-1. Northern pig-tailed macaques(M. leonina) are distributed in China and other surrounding Southeast Asia countries. Although northern pig-tailed macaques have been bred on a large scale as experimental animals since 2012, the reference value of normal levels of leukocytes is not available. To obtain such information, 62 blood samples from male and female healthy northern pig-tailed macaques at different ages were collected. The normal range of major leukocyte subpopulations, such as T lymphocytes, B lymphocytes, natural killer(NK) cells, monocytes, and the expression levels of activation or differentiation related molecules(CD38, HLA-DR, CCR5, CD21, IgD, CD80 and CD86) on lymphocytes were analyzed by flow cytometry. The counts of B cells decreased with age, but those of CD8+ T cells and NK cells and the frequency of CD38+HLA-DR+CD4+ T cells were positively correlated with age. The counts of leukocyte subpopulations were higher in males than those in females except for CD4+ T cells. Males also showed higher expression levels of Ig D and CD21 within B cells. This study provides basic data about the leukocyte subpopulations of northern pig-tailed macaques and compares this species with commonly used Chinese rhesus macaques(M. mulatta), which is meaningful for the biomedical application of northern pig-tailed macaques.
基金supported by the National Natural Science Foundation of China(81930071,82072502)the National Natural Science Foundation Regional Innovation and Development Joint Fund(U21A20352)+5 种基金the National Key Research and Development Project(2022YFC3601900,2022YFC2505500)the Project Program of National Clinical Research Center for Geriatric Disorders(Xiangya Hospital,2021LNJJ06,2022LNJJ07)the Natural Science Foundation of Hunan Province(2022JJ20100)the Key Research and Development Program of Hunan Province(2021SK2017)the Science and Technology Innovation Program of Hunan Province(2022RC3075)the Central South University Innovation-Driven Research Program(2023CXQD031)。
文摘Hand osteoarthritis is a common heterogeneous joint disorder with unclear molecular mechanisms and no disease-modifying drugs.In this study,we performed single-cell RNA sequencing analysis to compare the cellular composition and subpopulationspecific gene expression between cartilage with macroscopically confirmed osteoarthritis(n=5)and cartilage without osteoarthritis(n=5)from the interphalangeal joints of five donors.Of 105142 cells,we identified 13 subpopulations,including a novel subpopulation with inflammation-modulating potential annotated as inflammatory chondrocytes.Fibrocartilage chondrocytes exhibited extensive alteration of gene expression patterns in osteoarthritic cartilage compared with nonosteoarthritic cartilage.Both inflammatory chondrocytes and fibrocartilage chondrocytes showed a trend toward increased numbers in osteoarthritic cartilage.In these two subpopulations from osteoarthritic cartilage,the ferroptosis pathway was enriched,and expression of iron overload-related genes,e.g.,FTH1,was elevated.To verify these findings,we conducted a Mendelian randomization study using UK Biobank and a population-based cross-sectional study using data collected from Xiangya Osteoarthritis Study.Genetic predisposition toward higher expression of FTH1 mRNA significantly increased the risk of hand osteoarthritis(odds ratio=1.07,95%confidence interval:1.02–1.11)among participants(n=332668)in UK Biobank.High levels of serum ferritin(encoded by FTH1),a biomarker of body iron overload,were significantly associated with a high prevalence of hand osteoarthritis among participants(n=1241)of Xiangya Osteoarthritis Study(P-for-trend=0.037).In conclusion,our findings indicate that inflammatory and fibrocartilage chondrocytes are key subpopulations and that ferroptosis may be a key pathway in hand osteoarthritis,providing new insights into the pathophysiology and potential therapeutic targets of hand osteoarthritis.
基金supported by the grants from the Foundation of Science and Technology Department of Zhejiang Province for Beneficial Technology Research of Social Development(2011C23013)
文摘2,4-dinitrophenol (DNP), an organic compound which frequently used in industry, is considered to have high toxicity. This study aimed to investigate the early changes of lymphocyte subpopulations in patients with occupational 2,4-DNP poisoning. Totally 9 patients with acute occupational 2,4-DNP poisoning and 30 healthy volunteers as control were enrolled. The patients received immediately comprehensive supportive treatments, including large-dose glucocorticoid and repeated hemoperfusion (HP). The ratio of CD4+/CD8+ T cells were significantly higher in patients upon admission compared to healthy controls (P 〈 0.01); however, counts of total lymphocytes, CD3+, CD3+CD4+, CD3+CD8+, B (CD19+), and natural killer (NK) cells (CD16+CD56+) were significantly reduced (all P 〈 0.001). The NK cell count was negatively correlated with initial plasma 2,4-DNP concentration (r = -0.750, P = 0.026). Thus, acute occupational 2,4-DNP poisoning was accompanied by immediate complex immune cell reactions, especially NK cells might play important role in severe 2,4-DNP poisoning.
文摘This study was designed to analyze the sperm kinematic and morphometric subpopulations in the different fractions of the ejaculate in normozoospermic men. Ejaculates from eight normozoospermic men were collected by masturbation in three fractions after 3-5 days of sexual abstinence. Analyses of sperm motility by computer-assisted sperm analysis (CASA-Mot), and of sperm morphometry by computer-assisted sperm morphometry analysis (CASA-Morph) using fluorescence were performed. Clustering and discriminant procedures were performed to identify sperm subpopulations in the kinematic and morphometric data obtained. Clustering procedures resulted in the classification of spermatozoa into three kinematic subpopulations (slow with low ALH [35.6% of all motile spermatozoa], with circular trajectories [32.0%], and rapid with high ALH [32.4%]), and three morphometric subpopulations (large-round [33.9% of all spermatozoa], elongated [32.0%], and small [34.10%]). The distribution of kinematic sperm subpopulations was different among ejaculate fractions (P 〈 0.001), with higher percentages of spermatozoa exhibiting slow movements with low ALH in the second and third portions, and with a more homogeneous distribution of kinematic sperm subpopulations in the first portion. The distribution of morphometric sperm subpopulations was also different among ejaculate fractions (P〈 0.001), with more elongated spermatozoa in the first, and of small spermatozoa in the third, portion. It is concluded that important variations in the distribution of kinematic and morphometric sperm subpopulations exist between ejaculate fractions, with possible functional implications.
基金supported by a Perry Cross Spinal Research Foundation grant to JSJan Australian Research Council Discovery Grant DP150104495 to JE and JSJ
文摘OECs for spinal cord repair: Is repairing the iniured spinal cord by olfactory ensheathing cell (OEC) transplantation pos- sible? A recent human trial in which a paralysed man regained some function after transplantation of partially purified OECs suggests that this therapy may be a successful approach (Ta- bakow et al., 2014). In another human trial in which olfactory mucosa lamina propria was transplanted, patients recovered some motor and sensory function (Wang et al., 2015). While these results show promise, it is clear that improvements are needed to provide patients with increased functional output. Strategies to improve the therapeutic use of OECs may include improving the purification of the OECs used for transplantation, using them in combination with growth factors to combat the inhibitory environment and improve anon growth, the use of nerve bridges, advanced physiotherapy and the use of exo- skeleton robotics to reinforce functional connections. Of all these approaches, it is probably is primarily addressed to ensure crucial that the purity of OECs consistency in outcomes.
文摘Teratozoospermia (〈40% morphologically normal spermatozoa/ejaculate) is a frequent phenomenon in feline species. This research was carried out to study the possible differences in testicular volume, differential sperm morphometric traits, and potential differences regarding the sperm subpopulational structure during epididymal sperm maturation in teratozoospermic feline donors. Epididymal sperm samples were collected from the caput (R1), corpus (R2), and cauda (R3) epididymidis in two donor groups (N: normozoospermic; T: teratozoospermic). Aliquots were assessed for concentration, viability, motility, and acrosomal integrity. Sperm morphometric descriptors from CASA-Morph analysis were analyzed by the Principal Component Analysis (PCA) and clustering analyses. Irrespective of the group analyzed, PCA revealed two Principal Components (PCs) for each epididymal region explaining more than the 93% of the variance. Surprisingly, the number of subpopulations remained constant in regions R1-R2-R3 irrespective of the donor group analyzed. However, the distribution of these subpopulations was found to be structurally different and strongly influenced by the epididymal region and the donor group. In conclusion, testicular morphometry and the sperm subpopulation structure were different in N and T donors. The alterations in subpopulations during epididymal maturation could be used as a potential clinical indicator of teratozoospermic individuals since an important influence of teratozoospermia on sperm subpopulation structure has been demonstrated.
基金Supported by JSPS KAKENHI(Grants-in-Aid for Scientific Research) for Nishikawa S,No.26893172
文摘AIMTo identify and characterize functionally distinct subpopulation of adipose-derived stem cells (ADSCs). METHODSADSCs cultured from mouse subcutaneous adipose tissue were sorted fluorescence-activated cell sorter based on aldehyde dehydrogenase (ALDH) activity, a widely used stem cell marker. Differentiation potentials were analyzed by utilizing immunocytofluorescece and its quantitative analysis. RESULTSApproximately 15% of bulk ADSCs showed high ALDH activity in flow cytometric analysis. Although significant difference was not seen in proliferation capacity, the adipogenic and osteogenic differentiation capacity was higher in ALDH<sup>Hi</sup> subpopulations than in ALDH<sup>Lo</sup>. Gene set enrichment analysis revealed that ribosome-related gene sets were enriched in the ALDH<sup>Hi</sup> subpopulation. CONCLUSIONHigh ALDH activity is a useful marker for identifying functionally different subpopulations in murine ADSCs. Additionally, we suggested the importance of ribosome for differentiation of ADSCs by gene set enrichment analysis.
文摘Sperm quality is evaluated for the calculation of sperm dosage in artificial reproductive programs. The most common parameter used is motility, but morphology has a higher potential as a predictor of genetic quality. Morphometry calculations from CASA-Morph technology improve morphological evaluation and allow mathematical approaches to the problem. Semen from 28 Holstein bulls was collected by artificial vagina, and several ejaculates were studied. After general evaluation, samples were diluted, packaged in 0.25 ml straws, and stored in liquid nitrogen. Two straws per sample were thawed, and slides were processed and stained with Diff-Quik. Samples were analyzed by a CASA-Morph system for eight morphometric parameters. In addition to the "classical" statistical approach, based on variance analysis (revealing differences between animals, ejaculates, and straws), principal component (PC) analysis showed that the variables were grouped into PC1, related to size, and PC2 to shape. Subpopulation structure analysis showed four groups, namely, big, small, short, and narrow from their dominant characteristics, representing 31.0%, 27.3%, 24.1%, and 17.7% of the total population, respectively. The distributions varied between animals and ejaculates, but between straws, there were no differences in only four animals. This modern approach of considering an ejaculate sperm population as divided into subpopulations reflecting quantifiable parameters generated by CASA-Morph systems technology opens a new view on sperm function. This is the first study applying this approach to evaluate different ejaculates and straws from the same individual. More work must be done to improve seminal dose calculations in assisted reproductive programs.
文摘This study was designed to characterize morphometric sperm subpopulations in normozoospermic men by using different statistical methods and examining their suitability to classify correctly different sperm nuclear morphologies present in human ejaculates. Ejaculates from 21 normozoospermic men were collected for the study. After semen collection and analysis, samples were prepared for morphometric determination. At least 200 spermatozoa per sample were assessed for sperm morphometry by computer-assisted sperm morphometry analysis (CASA-Morph) using fluorescence. Clustering and discriminant procedures were performed to identify sperm subpopulations from the morphometric data obtained. Clustering procedures resulted in the classification of spermatozoa into three morphometric subpopulations (large-round 30.4%, small-round 46.6%, and large-elongated 22.9%). In the second analysis, using discriminant methods, the classification was made independently of size and shape. Three morphological categories according to nuclear size (small 〈10.90 μm^2, intermediate 10.91-13.07 μm^2, and large 〉13.07 μm^2) and four categories were defined on 400 canonical cells (100 × 4) from 10 men according to sperm nuclear shape (oval, pyriform, round, and elongated). Thereafter, the resulting classification functions were used to categorize 4200 spermatozoa from 21 men. Differences in the class distribution were observed among men from both clustering and discriminant procedures. It was concluded that the combination of CASA-Morph fluorescence-based technology with multivariate cluster or discriminant analyses provides new information on the description of different morphometric sperm subpopulations in normal individuals, and that important variations in the distribution of morphometric sperm subpopulations may exist between men, with possible functional implications.
文摘A study of the genetic diversity within four subpopulations of Chimonobambusa rivularis Yi and adaptations to altitude, habitat and related factors in Qionglai City of Sichuan province in China, analyzed by random amplified polymorphic DNA technique. 22 random primers were selected in the amplification and 375 repetitive loci with 350 polymorphic loci were produced. The total average percentage of amplification loci was 93.3%. The genetic diversity of every subpopulation was medium on (the percentage of amplification loci was 49.33%-66.67%). The genetic diversity of high altitude samples of Zhengtiantai was lower than that of low altitude Wutonggang's. There was no obvious differentiation between two subpopulations of Zhengtiantais. The correlation is very low between altitude and the amplification loci and the genetic distance, Pearson correlation coefficient was 0.431 and 0.488 (P 〈 0.01). Through investigation, the primary cause of heredity multiplicity drops was that the habitat had been destroyed by tourism development and other human disturbance.
文摘This work provides information on the blue fox ejaculated sperm quality needed for seminal dose calculations. Twenty semen samples, obtained by masturbation, were analyzed for kinematic and morphometric parameters by using CASA-Mot and CASA-Morph system and principal component (PC) analysis. For motility, eight kinematic parameters were evaluated, which were reduced to PC1, related to linear variables, and PC2, related to oscillatory movement. The whole population was divided into three independent subpopulations: SP1, fast cells with linear movement; SP2, slow cells and nonoscillatory motility; and SP3, medium speed cells and oscillatory movement. In almost all cases, the subpopulation distribution by animal was significantly different. Head morphology analysis generated four size and four shape parameters, which were reduced to PC 1, related to size, and PC2, related to shape of the cells. Three morphometric subpopulations existed. SPI: large oval cells; SP2: medium size elongated cells; and SP3. small and short cells. The subpopulation distribution differed between animals. Combining the kinematic and morphometric datasets produced PC1, related to morphometric parameters, and PC2, related to kinematics, which generated four sperm subpopulations - SPI.. high oscillatory motility, large and short heads; SP2; medium velocity with small and short heads; SP3. slow motion small and elongated cells; and SP4. high linear speed and large elongated cells. Subpopulation distribution was different in all animals. The establishment of sperm subpopulations from kinematic, morphometric, and combined variables not only improves the well-defined fox semen characteristics and offers a good conceptual basis for fertility and sperm preservation techniques in this species, but also opens the door to use this approach in other species, included humans.
基金Supported by the National Natural Science Foundation of China(61139002,61171132)the Natural Science Foundation of Jiangsu Education Department(12KJB520013)+2 种基金the Fundamental Research Funds for the Central Universitiesthe Funding of Jiangsu Innovation Program for Graduate Education(CXZZ110219)the Open Project Program of State Key Lab for Novel Software Technology in Nanjing University(KFKT2012B28)
文摘Attribute reduction is an important process in rough set theory.Finding minimum attribute reduction has been proven to help the user-oriented make better knowledge discovery in some cases.In this paper,an efficient minimum attribute reduction algorithm is proposed based on the multilevel evolutionary tree with self-adaptive subpopulations.A model of multilevel evolutionary tree with self-adaptive subpopulations is constructed,and interacting attribute sets are better decomposed into subsets by the self-adaptive mechanism of elitist populations.Moreover it can self-adapt the subpopulation sizes according to the historical performance record so that interacting attribute decision variables are captured into the same grouped subpopulation,which will be extended to better performance in both quality of solution and competitive computation complexity for minimum attribute reduction.The conducted experiments show the proposed algorithm is better on both efficiency and accuracy of minimum attribute reduction than some representative algorithms.Finally the proposed algorithm is applied to magnetic resonance image(MRI)segmentation,and its stronger applicability is further demonstrated by the effective and robust segmentation results.
文摘The main aims of this research were to study possible differences in objective morphometric sperm characteristics, establish normative sperm morphometry standards, and evaluate the presumed different subpopulation distribution of avian spermatozoa from the rooster (Gallus domesticus) and Guinea fowl (Numida meleagris) as model avian species. Seventy-two ejaculates (36 per species studied) were obtained manually, following a training period involving gently combined dorso-abdominal and lumbo.sacral massage of the birds. Ejaculates were processed for volume, sperm concentration, viability, motility, and morphology. Moreover, samples were submitted for sperm morphometric assessment using objective Computer-Assisted Semen Analysis for Morphometry (CASA-Morph) methods, with sperm morphometric descriptors evaluated by Principal Component Analysis (PCA) and multivariate clustering analyses. There were several differences observed between the avian species in values obtained for ejaculate volume and sperm concentration (P 〈 0.001). Irrespective of species, PCA revealed two Principal Components (PCs) explaining more than 80% of the variance. In addition, the number of subpopulations differed with species (three and five subpopulations for rooster and Guinea fowl, respectively). Moreover, the distribution of the sperm subpopulations was found to be structurally different between species. In conclusion, our findings from using CASA-Morph methods indicate pronounced sperm morphometric variation between these two avian species. Because of the strong differences observed in morphometric parameter values and their subpopulation distribution, these results suggest that application of objective analytical methods such as CASA-Morph could substantially improve the reliability of comparative studies and help establish valid normative sperm morphological values for avian species.
文摘This study aimed to identify and characterize the sperm subpopulations existing in water buffalo semen using a computer assisted sperm analyzer (CASA), as well as assess the effects of cryopreservation on the sperm subpopulation structure and evaluate bull variability. The semen of eight Bulgarian Murrah bulls was collected by four times in an interval of one week each. The semen was cryopreserved following a standard protocol and sperm kinematics was assessed. Clustering methods were applied to individual sperms, forming two significantly different (P 〈 0.05) subpopulations (P1 and P2). Subpopulation P1 represents those spermatozoa that moved most rapidly and progressively (46.29%), and subpopulation P2 includes spermatozoa with relatively low velocity or poorly motile but with high progressiveness (53.41%). There was a decline on the population of P1 sperms from fresh (52.52%), pre-freeze (45.73%) to post-thaw (35.17%) stages and significant difference on the sperm kinematics between P1 and P2. A significant decline in the values of distance, velocity and amplitude of lateral head (ALH) parameters were observed at post-thaw stage, while an increase was observed on trajectory and beat cross frequency (BCF) kinematics. Values of sperm kinematics were also significantly different (P 〈 0.05) among all bulls. The frequency distribution of spermatozoa on both subpopulations P1 and P2 was quite similar for all bulls in pre-freeze and post-thaw stages, but with significant (P 〈 0.05) variability on fresh stage. Bulls with the highest maintained frequency of P1 sperms are denoted as good freezer bulls. In sum, kinematic characterization of water buffalo sperm and clustering into subpopulation enable to identify bulls that are more resistant to cryopreservation and production of quality semen for genetic propagation.
文摘Objective: To study the cellular immunity function of patientswith early syphilis and the effects on immune modifiersEsberitox N or IFN. Methods: T-lymphocyte subpopulations of the peripheralblood in 44 patients with syphilis and 40 healthy controls wereexamined by flow cytometry. Results: The number of CD_4+ cells and the CD_4+/CD_8+ ratioin patients with syphilis were found to be significantly lowerthan those in the control (P<0.0l), while the number of CD_8+cells was higher than that in the control (P<0.01). TheCD_4+/CD_8+ ratio in those with active disease was lower thanthat in those who had been cured (P<0.05). The CD_4+ countand the CD_4+/CD_8+ ratio in those treated with antibiotics alone(Penicillin G or Cephalosporins) were lower than those treatedwith both antibiotics and immunomodulators (P<0.05). Conclusions: Cellular immunity in the patients with earlysyphilis was prominently suppressed, and treatment withimmunomodulators may be helpful for the recovery of cellularimmunity of these patients.
文摘<span style="font-family:Verdana;"><strong>Rationale</strong></span><b><span style="font-family:Verdana;">:</span></b><span style="font-family:Verdana;"> The known prognosis factors for rhabdomyosarcoma (RMS) in</span><span style="font-family:;" "=""> <span style="font-family:Verdana;">children do not always explain the unsatisfactory outcome of treatment.</span><span style="font-family:Verdana;"> Changes in the subpopulation composition of Bone Marrow (BM) effector cells during </span><span style="font-family:Verdana;">the development of RMS may indicate new directions for the search for </span><span style="font-family:Verdana;">prog</span><span><span style="font-family:Verdana;">nostic factors and points for the impact of targeted therapy. </span><b><span style="font-family:Verdana;">Purpose</span></b></span></span><b><span style="font-family:Verdana;">:</span></b><span style="font-family:Verdana;"> To</span><span style="font-family:;" "=""><span style="font-family:Verdana;"> identify correlations between quantitative changes in the levels of subpopulations of T, B and NK-lymphocytes of BM and known risk factors for RMS in child</span><span><span style="font-family:Verdana;">ren. </span><b><span style="font-family:Verdana;">Objects</span></b></span></span><b><span style="font-family:Verdana;">:</span></b><span style="font-family:Verdana;"> The study included 31 patients. The main group included 16</span><span style="font-family:;" "=""><span style="font-family:Verdana;"> patients with RMS, average age—6.8 ± 1.0 years, while children 1 - 10 years old—13 (81.3%), over 10 years old—3 (18.8%) people, girls and boys were 8 </span><span style="font-family:Verdana;">people each. The embryonic variant of RMS was established in 10 (62.5%) </span><span style="font-family:Verdana;">cases, the alveolar variant—in 4 (25%) cases, in two patients (12</span></span><span style="font-family:Verdana;">.</span><span style="font-family:;" "=""><span style="font-family:Verdana;">5%), the histological variant was not established. In 12 (75%) patients, an unfavorable loca</span><span style="font-family:Verdana;">lization of the RMS (parameningeal, extremities, prostate, bladder) was re</span><span style="font-family:Verdana;">vealed, in 4 patients (25%), the localization of </span></span><span style="font-family:Verdana;">the </span><span style="font-family:;" "=""><span style="font-family:Verdana;">tumor was regarded as favorable. Patients with T2b—13 (81.2%) and T2a—2 (12.5%) stages prevailed. Regional and distant metastases were detected in 10 (52.6%) patients. The comparison group included 15 children in whom the presence of malignant neoplasia was excluded, the average age was 8.4 ± 1.5 years, 11 boys (73.7%) and 4 girls </span><span><span style="font-family:Verdana;">(26.3%). </span><b><span style="font-family:Verdana;">Methods</span></b></span></span><b><span style="font-family:Verdana;">:</span></b><span style="font-family:Verdana;"> All patients underwent morphological (myelogram</span><span style="font-family:;" "=""><span style="font-family:Verdana;"> count</span><span style="font-family:Verdana;">ing) and immunological (quantitative analysis of lymphocytic subpopula</span><span style="font-family:Verdana;">tions) bone marrow studies. Immunophenotyping in all patients was carried out by direct immunofluorescence using a triple fluorescent label. </span><b><span style="font-family:Verdana;">Results</span></b></span><b><span style="font-family:Verdana;">:</span></b><b><span style="font-family:;" "=""> </span></b><span style="font-family:;" "=""><span style="font-family:Verdana;">Significant differences in the levels of subpopulations of BM T-lymphocytes were found when comparing the values of the main group, distributed by localization and histological variant, with the data obtained in the control group of patients. For example, the percentage of </span><span style="font-family:Verdana;">CD3+ T cells with the co-stimulatory molecule CD28+ was significantly higher in patients with parameningeal </span><span style="font-family:Verdana;">RMS </span><span><span style="font-family:Verdana;">(p = 0.010). </span><b><span style="font-family:Verdana;">Conclusion</span></b></span></span><b><span style="font-family:Verdana;">:</span></b><span style="font-family:Verdana;"> Each clinical group of patients has its own individual immunological characteristics. The results obtained by us can be considered indicative and regarded as starting points for further study of the peculiarities of the subpopulation composition of BM in patients with RMS.</span>