The composition of the intestinal microbiota after allogeneic haematopoietic stem cell transplantation and its association with graft versus host disease as assessed by 16Sribosomal ribonucleic acid.

To observe the evolution of the intestinal microbiota in patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and discuss the relationship between the intestinal microbiota and graft-versus-host disease (GVHD). In this study, 11 patients who underwent allo-HSCT in the Aerospace Central Hospital from January 2021 to October 2021 were selected, along with 11 donors. Fecal specimens were collected 7 times: at admission, after pre-treatment, and every 3 weeks after transplantation from patients and once from donors. The composition of the intestinal microbiota and its association with GVHD after allogeneic hematopoietic stem cell transplantation were analyzed by 16S rRNA sequencing. Of the 11 patients, 5 developed GVHD, and 6 did not. The diversity of the intestinal microbiota among GVHD patients first increased and then decreased after transplantation, while that among non-GVHD patients first increased and then tended to be stable. The diversity of the intestinal microbiota among GVHD patients was lower than that among non-GVHD patients before pre-treatment and after transplantation. The taxa diversity of the intestinal microbiota in the non-GVHD group was better than that in the GVHD group before allo-HSCT, and the difference was statistically significant (P<0.05 for OTUs and CHAO1 index). The taxa abundance of Enterococcaceae 2.16% (2.13%, 2.22%) before allo-HSCT was significantly higher than that in the non-GVHD group 1.33% (0.27%, 1.52%), and the difference was statistically significant (P=0.004). There was no significant difference between the GVHD group and the non-GVHD group in the diversity of the intestinal microbiota of donors (P<0.05). The characteristics of the intestinal microbiota in the final sample of patients in the GVHD group were similar to the preoperative structure of the intestinal microbiota. In conclusion: The decrease in the diversity of the intestinal microbiota after HSCT may be a risk factor for the occurrence of GVHD. The presence of Enterococcaceae in the intestinal microbiota may be associated with an increased risk of developing GVHD. The intestinal microbiota reconstitute to be close to the intestinal microbiota composition of the donors in the non-GVHD group.

Alprostadil alleviates liver injury in septic rats via TLR4/NF-κB pathway.

OBJECTIVE: The aim of this study was to explore the role of alprostadil (Alp) in cecal ligation and puncture (CLP)-induced septic injury in rats and its possible mechanism of action. MATERIALS AND METHODS: Wistar rats were randomly assigned into three groups, including: Sham group (no CLP was performed), CLP group (CLP was conducted) and Alp group (Alp was injected after CLP). Serum liver function markers, pathological changes in liver tissues, alterations in the level of oxidative stress, activity of the Toll-like receptor 4 (TLR4)/nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway, and release of inflammatory factor tumor necrosis factor alpha (TNF-α) in the liver tissue homogenate were detected in each group. RESULTS: Compared with Sham group, the rats in CLP group had substantially elevated content of serum liver function markers, increased apoptotic liver cells, upregulated levels of oxidative stress, enhanced activity of the TLR4/NF-κB pathway, and increased release of TNF-α (p<0.05). Meanwhile, there were evident pathological changes under microscopic examination in CLP group compared with Sham group (p<0.05). In comparison with CLP group, Alp group exhibited significantly decreased concentrations of liver function markers, microscopic findings, such as decreased inflammatory cell infiltration in the interstitum, notably lowered proportion of apoptotic cells, decreased level of oxidative stress, weakened activity of the TLR4/NF-κB pathway and restrained release of TNF-α (p<0.05). Furthermore, normal morphology of liver cells was observed in Alp group compared with CLP group (p<0.05). CONCLUSIONS: Alp alleviates liver injury in septic rats by inhibiting the TLR4/NF-κB pathway.

Profiling of miRNAs in neonatal cloned bovines with collapsed lungs and respiratory distress.

Neonatal respiratory distress is a major mortality factor in cloned animals; however, the pathogenesis of this disease has rarely been investigated. Previous studies have shown that miRNAs regulate critical genes related to lung development, cell differentiation, surfactant synthesis, secretion and lung disease. This study aimed to examine differentially expressed miRNAs in collapsed lungs of cloned bovine neonates and normal lungs in order to identify key pathways and functions that might be related to the pathogenesis of neonatal respiratory distress. In this study, miRNA transcriptomes of collapsed lungs of neonatal cloned bovines and normal lungs were analysed by next-generation sequencing and the results were validated using quantitative real-time PCR (qRT-PCR). A total of 177 differentially expressed miRNAs were identified in the two groups (fold change > 2, RPM ≥ 5), some of which were associated with type II cell differentiation, for example, mmu-miR-29a-5p_L-2R+1, hsa-miR-200c-5p_L-1R+1 and mmu-miR-18a-3p_R+1. The differentially expressed miRNAs were predicted to 6,031 target genes. By Gene Ontology (GO) and Kyoto Encyclopeida of Genes and Genomes (KEGG) DATA base, 133 significant GO terms (p < .05) and 13 significant KEGG pathways (p < .05) were obtained. Many of them were associated with lung development and surfactant homoeostasis, such as lipid biosynthetic processes, protein transport, endocytosis, lysosome, endosome, Golgi apparatus and membrane. Our results of miRNAs express profiles may partially explain the respiratory distress and lung collapse in neonatal bovine clones and could provide novel insights into roles of miRNAs in regulation of lung collapse and neonatal respiratory distress in cloned farm animals.

Effect of vitrification on the mRNA transcriptome of bovine oocytes.

Vitrification has been shown to decrease the developmental capacity of mammalian oocytes, and this is closely associated with the abnormal mRNA expressions of vitrified oocytes. However, the effect of vitrification on transcriptional machinery of oocytes examined by RNA sequencing (RNA-seq) has yet to be defined. In the present study, the mRNA transcriptomes of fresh and vitrified bovine oocytes were analysed by Smart-seq2 with the differently expressed genes determined by DEseq2 (an adjusted p-value of .05 and a minimum fold change of 2). The differentially expressed mRNAs were then searched against the Gene Ontology (GO) and Genomes (KEGG) database. Finally, the mRNA expressions of 10 candidate genes were validated using quantitative real-time PCR (qRT-PCR). Approximately 12,000 genes were detected in each sample of fresh or vitrified oocytes. Of these, the expression levels of 102 genes differed significantly in vitrified groups: 12 genes mainly involved in cell cycle, fertilization and glucose metabolism were upregulated, and 90 genes mainly involved in mitochondria, ribosomal protein, cytoskeleton, transmembrane protein, cell cycle and calcium ions were downregulated. GO analysis showed that these genes were mainly enriched in terms of membrane-bounded organelles, macromolecular complex, and intracellular part. The mRNA expression levels of 10 candidate genes selected randomly were in agreement with the results of the RNA-seq. In conclusion, our results showed that vitrification affected the mRNA transcriptome of bovine oocytes by downregulating genes, which contributed to the decreased developmental capacity of vitrified oocytes. Our findings will be useful in determining approaches to improve the efficiency of vitrified oocytes.

Association between -1082G/A, -819C/T, and -592C/A genetic polymorphisms in IL-10 and risk of type 2 diabetes mellitus in a Chinese population.

Type 2 diabetes mellitus is the most common form of endocrine disease in humans; genetic factors are known to contribute to the development of this disease. In this case-control study, we investigated the relationship between the -1082G/A, -819C/T, and -592C/A polymorphisms in interleukin 10 (IL-10) and the pathogenesis of type 2 diabetes mellitus in a Chinese population. Patients with type 2 diabetes mellitus (N = 228) and control subjects (N = 240) were recruited from the Department of Endocrinology at the People's Hospital of Linyi City, between September 2013 and April 2015. The IL-10 -1082G/A, -819C/T, and -592C/A polymorphisms were genotyped by polymerase chain reaction-restriction fragment length polymorphism. Multivariate logistic regression analyses revealed that patients carrying the AA genotype of IL-10 -592C/A were at a higher risk of developing type 2 diabetes mellitus compared to those carrying the CC genotype [adjusted odds ratio (OR) = 1.74; 95% confidence interval (CI) = 1.03-2.95]. In addition, individuals carrying the A allele of IL-10 -592C/A showed a 1.34-fold higher risk of developing type 2 diabetes mellitus compared to those carrying the C allele (adjusted OR = 1.34; 95%CI = 1.03- 1.75). There was no significant correlation between the IL-10 -1082G/ A and -819C/T polymorphisms and risk of type 2 diabetes mellitus. In conclusion, this study shows that the -1082G/A polymorphism of IL-10 contributes to the onset of type 2 diabetes mellitus, and may be considered a biomarker for early screening of type 2 diabetes mellitus in the Chinese population studied here.

Transcriptome analyses of inner cell mass and trophectoderm cells isolated by magnetic-activated cell sorting from bovine blastocysts using single cell RNA-seq.

Research on bovine embryonic stem cells (bESCs) has been hampered because bESCs are cultured in conditions that are based on information obtained from culturing mouse and human inner cell mass (ICM) cells. The aim of this study was to compare gene expression in ICM and trophectoderm (TE) cell lineages of bovine embryos and to discuss the findings relative to information available for mice and humans. We separated a high-purity (>90%) ICM and TE from bovine blastocysts by magnetic-activated cell sorting and analysed their transcriptomes by single cell RNA-seq. Differentially expressed genes (DEGs) were assessed using Gene Ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) databases. Finally, qRT-PCR was performed to validate the RNA-seq results. From 207 DEGs identified (adjusted p ≤ .05; fold change ≥2), 159 and 48 had greater expression in the ICM and TE cells respectively. We validated 27 genes using qRT-PCR and found their expression patterns were mostly similar to those of RNA-seq, including 12 novel ICM-dominant (HNF4A, CCL24, FGFR4, IFITM3, PTCHD2, GJB5, FN1, KLK7, PRDM14, GRP, FGF19 and GCM1) and two novel TE-dominant (SLC10A1 and WNT4) genes. Bioinformatics analysis showed that these DEGs are involved in many important pathways, such as MAPK and cancer cell pathways, and these pathways have been shown to play essential roles in mouse and human ESCs in the self-renewal and pluripotent maintenance. As a conclusion, there were sufficient differences to allow us to conclude that the control of pluripotency in bovine ICM cells is species-specific.

2D speckle tracking imaging to assess sepsis induced early systolic myocardial dysfunction and its underlying mechanisms.

OBJECTIVE: The aim of this study was to apply speckle tracking imaging (STI) to evaluate rabbit sepsis induced myocardial injury. MATERIALS AND METHODS: Rabbits were injected with the endotoxin lipopolysaccharide (LPS). before and 2h, 4h, 6h, 8h, 12h after the injection of LPS, conventional echocardiography and STI were performed to measure left ventricular end diastolic diameter (LVDd), end-systolic diameter (LVDs), cardiac output (CO), left ventricular ejection fraction (EF), fractional shortening (FS), left ventricular global longitudinal strain (GLS)/GLS rate (GLSr), global circumferential strain/strain rate (GCS/GCSr), and the global radial strain/strain rate (GRS/GRSr). We also observed the injury with optical microscopy. Serum troponin cTnT and TUNEL assays determining myocardial apoptosis were also carried out to determine the induced injury levels. RESULTS: None of the indicators measured are statistically different in the two groups before the LPS treatment (p > 0.05). 2h after LPS injection, in LPS treated group, only GLS/GLSr, GCS/GCSr significantly reduced compared to the control group (p < 0.05). 6h, 8h, 12h after LPS injection, EF and FS were lower (p < 0.05), LVDd and LVDs were higher (p < 0.05), and GLS/GLSr, GCS/GCSr, GRS/GRSr were significantly lower (p < 0.05). LPS significantly elevated serum cTnT level in the experimental group (p < 0.05). In the LPS treated group, we found pathological changes of the myocardium. CONCLUSIONS: STI is more sensitive than the conventional echocardiography for the early detection of abnormal myocardial contractility. It is a noninvasive, accurate and timely detection of myocardial dysfunction in sepsis.

Apoptosis-like events and in vitro fertilization capacity of sex-sorted bovine sperm.

This study utilized three staining assays (Annexin V, mitochondrial membrane potential (JC-1) and TUNEL) for flow cytometric analysis of apoptosis in sex-sorted sperm from four different bulls (A, B, C and D). Correlations between sperm quality and IVF efficiency were then assessed to determine which assay provided the best prediction of IVF efficiency. The results of the Annexin V assays, as well as measures of viable sperm, early apoptosis, necrotic sperm and mitochondrial membrane potential (∆ψm) showed that the sex-sorted sperm collected from bull A significantly differed from those of the other three bulls (p < 0.05). In addition, the levels of DNA fragmentation in sex-sorted sperm from bull A were significantly lower than those from bulls B and C (p < 0.05). The percentage of cells reaching the cleavage and blastocyst stages in sex-sorted sperm from bull A were significantly greater than those from the other bulls (p < 0.05). A significant positive correlation was observed between viable sperm and the percentage of cells at the cleavage or blastocyst stages (p < 0.05). In contrast, a negative correlation was found between early apoptotic sperm and the percentage of cells at the cleavage or blastocyst stages (p < 0.05). In conclusion, these results indicated that the Annexin V assay was the most reliable technique for the prediction of the IVF success of sex-sorted bovine sperm.

Immuno-inflammatory regulation effect of mesenchymal stem cell transplantation in a rat model of myocardial infarction.

BACKGROUND: Mesenchymal stem cells (MSC) have recently been shown to possess immunomodulatory properties in vitro and in vivo. The present study aimed to investigate the regulatory effect of MSC transplantation on the immuno-inflammatory response in myocardial infarction (MI). METHODS: MI was induced in Sprague-Dawley rats by left anterior descending coronary artery ligation, and the animals were randomly assigned into the following three groups: sham ( n=8); phosphate-buffered saline (PBS) injected (MI+PBS, n=8); and MSC transplantation (MI+MSC, n=8). BrdU-labeled MSC or PBS was transplanted into peri-infarct myocardium by direct myocardial injection. At 1 and 28 days post-transplantation, cardiac function was evaluated by echocardiography. Transplanted cells were investigated through immunohistochemistry. Lymphocyte cytotoxic activity was evaluated with the crystal violet method. The activity of NF-kappaB and protein expression of tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-6 and IL-10 in myocardium were assessed by immunohistochemistry and Western blot. RESULTS: Echocardiographic examination revealed that the MSC transplantation prevented left ventricular dilation and dysfunction at 28 days after the operation. BrdU-stained cells were found living in host heart 4 weeks after transplantation. MSC transplantation attenuated the cytotoxic activity of spleen lymphocytes. Transplantation of MSC inhibited the activity of NF-kappaB, attenuated the protein production of TNF-alpha and IL-6, and increased the expression of IL-10 in peri-infarct myocardium. DISCUSSION: MSC transplantation modulated the immuno-inflammatory response in MI. The immuno-inflammatory regulatory effect of MSC transplantation might partly account for the cardiac protection in myocardial infarction.

Follow-up analysis of 180 Chinese Han families: identification of a novel locus for psoriasis at 2p22.3-11.2.

BACKGROUND: Psoriasis is a common inflammatory and hyperproliferative skin disease. The pathogenesis of psoriasis remains obscure. Family and twin studies have suggested a strong genetic susceptibility to psoriasis. Eight linkage loci (PSORS1-7, PSORS9) were identified and accepted by the OMIM and an additional 16 susceptibility loci have been suggested so far. OBJECTIVES: To investigate further three suggested psoriasis susceptibility loci at 2p22.3-11.2, 13q21-32 and 17q22-25.3 in a Chinese population. Using an expanded sample of 180 Chinese families with psoriasis and improved marker coverage, we verified whether they were Chinese Han psoriasis susceptibility loci. METHODS: In total, 180 Chinese Han families with psoriasis vulgaris (including the 61 families used in the original genome-wide scan and 119 new families) were recruited from the Dermatology Department at the First Hospital Affiliated to Anhui Medical University. Two-point and multipoint parametric and nonparametric linkage (NPL) analyses were performed at 2p, 13q and 17q in the total 180 families as well as the 61 original and 119 new families separately. RESULTS: At the region 2p, a maximum multipoint NPL score of 4.11 was identified at locus D2S337 (P=0.000003), and a maximum multipoint heterogeneity LOD (HLOD) score of 4.93 (alpha=54%) was identified at the same locus in the analysis of the 180 families. However, the analysis of the 180 families did not identify any significant linkage evidence at the region 13q21-32 [a maximum multipoint HLOD score of 0.10 (alpha=7%) and NPL score of 0.95 (P=0.14)] or the region 17q22-25.3 [a maximum multipoint HLOD score of 0.08 (alpha=6%) and NPL value of 0.94 (P=0.14)]. For these two regions, the LOD scores from the 180 families as well as the 119 new families were much smaller than the ones obtained from the original 61 families. CONCLUSIONS: Our study indicates that 2p22.3-11.2 is a novel psoriasis susceptibility locus in the Chinese Han population and confirms that psoriasis is a genetically heterogeneous disease.

Childhood psoriasis: a study of 277 patients from China.

OBJECTIVES: Psoriasis is common in childhood. The aim of this study was to present the clinical and epidemiological profile of childhood psoriasis in China. METHODS: A total of 277 childhood psoriasis patients younger than 16 years old were enrolled. Statistical analysis and heritability were performed using EPI INFO 6.0, spss 10.0 and Falconer's method. RESULTS: The median age was 11 years. The male : female ratio was 1:1.13. The median age of onset was 10 years. Of the patients, 48.7% had previous episodes of psoriasis. Of the 277 children with psoriasis, 68.6% had plaque-type psoriasis, 28.9% had guttate psoriasis, 1.1% presented pustular forms of psoriasis and 1.4% had erythroderma. The extensor surface of the extremities was the most frequently affected site in our patients, followed by the appearance of lesions on the scalp. A positive family history of psoriasis was found in 34.3% patients. The prevalence of psoriasis in first- and second-degree relatives was 7.0% and 1.0%, respectively. The heritability of psoriasis in first- and second-degree relatives was 72.67% and 55.18%, respectively. CONCLUSION: Our epidemiologic studies offer the information about Han Chinese distribution, which provide clues to describe psoriasis in children.

The familial risk of acne vulgaris in Chinese Hans - a case-control study.

BACKGROUND: Acne is a chronic inflammatory disease of the pilosebaceous follicles. Recent studies bring us increasing evidences that hereditary factors play an important but indirect role in acne. OBJECTIVE: To investigate the possible role of genetic factors in the pathogenesis of acne vulgaris in Chinese Han ethnic group. PATIENTS AND METHODS: Volunteers of 975 acne cases and 580 controls were included, contributing 3009 and 1825 first-degree relatives, respectively. One thousand and eighty-five first-degree relatives of acne cases were affected with facial acne. This compared with 223 first-degree relatives of non-acne controls. The odds ratio was used to estimate the relative risk for acne vulgaris associated with having an affected first-degree relative. RESULTS: The risk of acne vulgaris occurring in a relative of a patient with acne vulgaris was significantly greater than for the relative of an unaffected individual (odds ratio 4.05, 95% confidence interval (CI): 3.45-4.76, P<0.001). CONCLUSION: Our study suggests that familial factors are important in determining individual susceptibility to acne vulgaris.

A novel missense mutation L468Q of keratin 6a in pachyonychia congenita type 1.

BACKGROUND: Pachyonychia congenita is an autosomal dominant disorder that usually develops in early infancy. The major features of the syndrome are hypertrophic nail dystrophy, palmoplantar keratoderma and oral leucokeratosis, accompanied by other ectodermal defects, according to subtype. OBJECTIVE: To analyse the K6a gene mutation in a sporadic Chinese patient with pachyonychia congenita type 1 (PC-1) and to explore the relationship between the genotype and phenotype of PC-1. METHODS: Genomic DNA was extracted from peripheral blood of the patient with PC-1 and 100 unrelated controls. The whole coding region of K6a gene was amplified using long-range polymerase chain reaction (PCR); nested PCR was then used to amplify the mutation 'hot-spot' of the K6a gene. The PCR products were directly sequenced to detect the mutation. RESULTS: A novel missense mutation L468Q in the helix 2B domain of the K6a polypeptide was identified in the patient but not in the healthy individuals from the family and 100 unrelated control individuals. CONCLUSIONS: We describe this mutation for the first time, and provide further evidence that the helix boundary motif sequences of K6a are a mutation 'hot-spot'.

Clinical profiles of vitiligo in China: an analysis of 3742 patients.

Very few articles have aimed to illuminate the clinical profiles of vitiligo in China. We conducted this retrospective survey involving 4118 outpatients with vitiligo in order to identify the differences among various clinical types of vitiligo and their associated disorders. Completed questionnaires (3742) were validated and analysed. Of this large cohort, 1565 (41.8%) individuals presented vitiligo vulgaris, followed by focal, segmental, acrofacial, and universal, in that order. The mean age of vitiligo onset was 18.88 years. More than 60% of the patients were affected before 20 years of age. Patients with segmental vitiligo were affected earlier than those with other types of vitiligo (15.55 years; (P < 0.001). More than 74% of the patients presented with focal vitiligo at onset. After 3-5 years, 99% of active vitiligo was worse and shifted from one clinical type to another. However, there was no transformation between acrofacial vitiligo and segmental vitiligo. Compared with the general population, the patients with vitiligo were more likely to be affected by rheumatoid arthritis (P < 0.01), ichthyosis (P < 0.01), chronic urticaria (P < 0.01), or alopecia areata (P < 0.01).

Refinement of a locus for Marie Unna hereditary hypotrichosis to a 1.1-cM interval at 8p21.3.

BACKGROUND: Marie Unna hereditary hypotrichosis (MUHH) is a rare autosomal congenital alopecia with progressive hair loss starting in early childhood and accelerating at puberty. A locus for MUHH has been mapped on chromosome 8p21 but no genes for MUHH have been identified to date. OBJECTIVES: To refine the MUHH locus to a narrow chromosome region to facilitate cloning of the gene. METHODS: We performed genotyping and linkage analysis in a multigeneration Chinese family with MUHH, using 18 high-density microsatellite markers spanning the previously mapped interval at 8p21. RESULTS: Significant evidence for linkage was observed in this region, with a maximum two-point LOD score of 3.01 (theta = 0). Haplotype analysis localized the MUHH locus within the region defined by D8S282 and D8S1839. This region overlaps by 1.1-cM with the previously reported MUHH region and represents a physical distance of about 380 kb. CONCLUSIONS: This study provides a refined map location (1.1 cM) for isolation of the gene causing MUHH. These data also indicate the existence of a common MUHH locus at 8p21.3 between affected caucasian and Chinese families.

[Clinical study of various growth factors on the improvement of impaired healing ulcers in patients with diabetic disease].

OBJECTIVE: To investigate clinical effects and possible mechanisms of various growth factors on impaired healing ulcers of patients with diabetic disease. METHODS: Seventy-eight patients were divided into three groups; saline control, epidermal growth factor(EGF) experimental group, and platelet-derived wound healing factor (PDWHF) experimental group. General healing conditions, wound closing index, healing rates and histological changes of the patient's ulcer wound were observed during 1-8 weeks after treatment. RESULTS: The wound closing index and healing rate of ulcers were significantly increased in the EGF and PDWHF experimental groups compared with the control group, while the angiogenesis, fibroblast hyperplasia, and collagen deposit were more obvious in EGF and PDWHF experimental groups than that of control group. The promoting effects on wound healing in PDWHF experimental group were better than in EGF group. CONCLUSION: It suggests that local application of certain growth factor alone or various growth factors together is an effective method to improve the condition of impaired healing of diabetic ulcers.