Identification of new single nucleotide polymorphisms affecting total number born and candidate genes related to ovulation rate in Chinese Erhualian pigs.

The Chinese Erhualian pig has the highest record for litter size in the world. However, the genetic mechanism of its high prolificacy remains poorly understood. In our study, large phenotypic variations in litter size were found among Erhualian sows. Significant differences in total number born (TNB) and corpora lutea numbers were observed between sows with high and low estimated breeding values (EBVs) for TNB. To identify single nucleotide polymorphisms (SNPs) associated with TNB, a selective genomic scan was conducted on 18 sows representing the top 10% and 18 sows representing the bottom 10% of EBVs of 177 sows using Illumina Porcine SNP60 genotype data. Genome-wide fixation coefficient (FST ) values were calculated for each SNP between the high- and low-EBV groups. A total of 154 SNPs were significantly differentiated loci between the two groups. Of the top 10 highest FST SNPs, rs81399474, rs81400131 and rs81405013 on SSC8 and rs81434499 and rs81434489 on SSC 12 corresponded to previously reported QTL for litter size. The other five SNPs, rs81367039 on SSC2, rs80891106 on SSC7, rs81477883 on SSC12 and rs80938898 and rs80971725 on SSC14, appeared to be novel QTL for TNB. Significant associations between rs81399474 on SSC8 and TNB were confirmed in 313 Erhualian sows. Forty genes were identified around the top 10 highest FST SNPs, of which UCHL1, adjacent to rs81399474, and RPS6KB1 and CLTC, adjacent to rs81434499, have been reported to affect the ovulation rate in pig. The findings can advance understanding of the genetic variations in litter size of pigs.

Cell proliferation and apoptosis in the fetal and neonatal ovary of guinea pigs.

The guinea pig is an excellent animal model for studying reproductive biology of adult humans and most domestic animals. Yet, whether this animal might serve as a good model for embryonic stage investigations and determinations of signals affecting or directing ovary development remains unknown. These questions were addressed by examining morphological evolution and the expression of biomarkers of cell proliferation and apoptosis in the ovaries of fetal and neonatal guinea pigs in the present study. Embryonic and neonatal guinea pigs at 30, 40, 50, 60, and 68 days postcoitum (dpc) and at 1 day postpartum (dpp) were evaluated, and the dynamic changes in follicles between 30 dpc and 1 dpp were observed. Results also showed that a critical period of follicular development in guinea pig embryos occurred at 40 to 50 dpc. Moreover, the proliferating-cell nuclear antigen, a cell proliferation marker, immunohistochemically stained healthy follicles, while caspase-3, an apoptosis marker, was mainly observed in atretic follicles. Together, these results demonstrate that cell proliferation and apoptosis contribute to follicular formation, development, and atresia in fetal and neonatal guinea pig ovaries. Furthermore, this study confirmed that the guinea pig is also an excellent animal model for studying reproductive biology in human and domestic animal embryos.

[Exploration and practice of novel models of cellular therapy and hematopoietic stem cell transplantation].

Hematopoietic stem cell transplantation provides an effective cure for various hematological diseases, especially malignant hematological diseases, its treatment system has been continuously optimized, the source of donors has been expanding, the indications have been expanding, and the therapeutic effect has also made breakthroughs to a certain extent. At present, the status of hematopoietic stem cell transplantation technology in most hematological diseases is still unshakable, but the recurrence of the primary disease and complications related to hematopoietic stem cell transplantation are still two major clinical challenges that affect the long-term survival and quality of life of patients. Cell therapy represented by chimeric antigen receptor T (CAR-T) has made breakthrough progress in the treatment of refractory/recurrent B-cell malignancies. Compared with traditional drugs, cell therapy has unique in vivo metabolic characteristics, relying on immune specific recognition and the repair ability of stem cells. It is currently emerging in the treatment of blood tumors and the management of transplant complications. Multiple clinical studies have preliminarily demonstrated a new diagnostic and therapeutic model combining cell therapy with hematopoietic stem cell transplantation.

[Evaluation of differences in quality of life in patients with chronic graft-versus-host disease].

Objective: To evaluate the status of, differences in, and factors influencing quality of life (QoL) in patients with chronic graft-versus-host disease (GVHD). Methods: From September 2021 to February 2023, a cross-sectional study of 140 patients with chronic GVHD was conducted at our center. Symptom burden was assessed by the Lee Symptomatology Scale (LSS), and QoL was assessed by the Medical Outcome Study 36-Item Short-Form Health Survey (SF-36) (version 1) and five-level EuroQoL five-dimensional questionnaire (EQ-5D-5L). Results: Data from 140 respondents, including 32 (22.9%) with mild chronic GVHD, 87 (62.1%) with moderate chronic GVHD, and 21 (15.0%) with severe chronic GVHD, were analyzed. Of the respondents, 61.4% were male, and the median transplantation age was 34 (15-68) years. The primary diagnoses were acute myeloid leukemia (50.0%), acute lymphoblastic leukemia (20.0%), and myelodysplastic syndrome (15.0%). The common chronic GVHD-affected organs included the skin in 74 patients (52.9%), the eyes in 57 patients (40.7%), and the liver in 50 patients (35.7%). Among the whole cohort, the eye (20.48±23.75), psychological (16.13±17.00), and oral (13.66±20.55) scores were highest in the LSS group. The physiological function (36.07±11.13), social function (36.10±10.68), and role-emotional functioning (38.36±11.88) scores were lowest in the SF-36 group. The EQ-5D index was 0.764. The total LSS scores for mild, moderate, and severe chronic GVHD were 6.51±6.15, 10.07±5.61, and 20.90±10.09, respectively. The SF-36 physical component scores (PCSs) were 43.12±6.38, 40.73±7.14, and 36.97±6.97, respectively, and the mental component scores (MCSs) were 43.00±8.47, 38.90±9.52, and 28.96±9.63, respectively. The EQ-5D values were 0.810±0.124, 0.762±0.179, and 0.702±0.198, respectively. The multivariate analysis showed that the overall symptom burden (ß=-0.517), oral symptom burden (ß=-0.456), National Institute of Health (NIH) criteria for the eyes (ß=-0.376), and nutrition-related symptom burden (ß=-0.211) were significantly negatively correlated with the PCS. The NIH score (ß=-0.260) was negatively correlated with the MCS score. Oral symptom burden (ß=-0.400), joint/fascia NIH criteria (ß=-0.332), number of involved systems (ß=-0.253), overall NIH criteria (ß=-0.205), and number of immunosuppressants taken (ß=-0.171) were significantly negatively correlated with the EQ-5D score (all P<0.05). Medium to strong correlations were found between the EQ-5D score and the SF-36 score (|r|=0.384-0.571, P<0.001). Conclusions: The QoL of patients with chronic GVHD is impaired, and the more severe the disease, the poorer the QoL. Overall symptom burden, severity of eyes, and oral symptom burden were the most important factors affecting QoL.

Effects of Dietary Chromium Methionine on Growth Performance, Carcass Composition, Meat Colour and Expression of the Colour-related Gene Myoglobin of Growing-finishing Pigs.

To investigate the effect of dietary chromium (Cr) as Cr methionine (CrMet) on growth performance, carcass traits, pork quality, meat colour and expression of meat colour-related genes in growing-finishing pigs, 189 crossbred Duroc×(Landrace×Yorkshire) growing-finishing pigs (male, castrated, average initial BW 74.58±1.52 kg) were selected and randomly allocated into four groups. Dietary treatments per kg of feed were as follows: 0 (CT), 0.3 mg/kg (T1), 0.6 mg/kg (T2) and 0.9 mg/kg (T3) Cr (in the form of CrMet; as-fed basis), and each treatment was replicated five times with 8 to 10 pigs per replicate pen. During the 28 d of the experiment, both the ADG and the ADFI increased linearly (p<0.05) as the level of dietary Cr increased. The F/G ratio decreased linearly (p<0.05). As dietary Cr increased, loin muscle areas (linear, p = 0.013) and average backfat thickness (linear, p = 0.072) decreased. Shear force (linear, p = 0.070) and Commission Internationale de I'Éclairage (CIE) redness (quadratic, p = 0.028) were increased. In addition, CIE Lightness (quadratic, p = 0.053) were decreased as dietary Cr increased. As dietary Cr increased, total myglobin (Mb) content (quadratic, p = 0.015) and the mb mRNA levels (quadratic, p = 0.046) in longissimus muscles of pigs were up-regulated. In conclusion, supplementation of dietary Cr improved growth and meat colour, but increased shear force and decreased IMF reduced palatability of longissimus muscles. Moreover, the increasing total Mb content and mb mRNA levels indicated that CrMet dietary supplementation may improve meat colour via up-regulating expression of the mb gene.

Genome-wide association study reveals a quantitative trait locus and two candidate genes on Sus scrofa chromosome 5 affecting intramuscular fat content in Suhuai pigs.

Intramuscular fat content (IFC) is an essential quantitative trait of meat, affecting multiple meat quality indicators. A certain amount of IFC could not only improve the sensory score of pork but also increase the flavour, tenderness, juiciness and shelf-life. To dissect the genetic determinants of IFC, two methods, including genome-wide efficient mixed-model analysis (GEMMA) and linkage disequilibrium adjusted kinships (LDAKs), were used to carry out genome-wide association studies for IFC in Suhuai pig population. A total of 14 and 18 significant single nucleotide polymorphisms (SNPs) were identified by GEMMA and LDAK, respectively. The results of these two methods were highly consistent and all 14 significant SNPs in GEMMA were detected by LDAK. Seven of the 18 SNPs reached the genome-wide significance level (P < 9.85E-07) while 11 cases reached the suggestive significance level (P < 1.77E-05). These significant SNPs were mainly distributed on Sus scrofa chromosome (SSC) 5, 3, and 7. Moreover, one locus resides in a 2.27 Mb (71.37-73.64 Mb) region on SSC5 harbouring 13 significant SNPs associated with IFC, and the lead SNP (rs81302978) also locates in this region. Linkage disequilibrium (LD) analysis showed that there were four pairs of complete LD (r2 = 1) among these 13 SNPs, and the remaining 9 SNPs with incomplete LD (r2 ≠ 1) were selected for subsequent analyses of IFC. Association analyses showed that 7 out of 9 SNPs were significantly associated with IFC (P < 0.05) in 330 Suhuai pigs, and the other 2 SNPs tended to reach a significant association level with IFC (P < 0.1). The phenotypic variance explained (PVE) range of these 9 SNPs was 0.92-3.55%. Meanwhile, the lead SNP was also significantly associated (rs81302978) with IFC (P < 0.05) in 378 commercial hybrid pigs (Pietrain × Duroc) × (Landrace × Yorkshire) (PDLY), and the PVE was 1.38%. Besides, two lipid metabolism-relevant candidate genes, the leucine rich repeat kinase 2 (LRRK2) and PDZ domain containing ring finger 4 (PDZRN4) were identified in the 2.27 Mb region on SSC5. In conclusion, our results may provide a set of markers useful for genetic improvement of IFC in pigs and will advance the genome selection process of IFC on pig breeding programmes.

Expression of genes involved in the somatotropic, thyrotropic, and corticotropic axes during development of Langshan and Arbor Acres chickens.

We investigated changes in mRNA expression of the somatotropic, thyrotropic, and corticotropic axes of Langshan (LS) and Arbor Acres (AA) broiler chickens during embryonic and postnatal development. We found an inverse expression profile between pituitary growth hormone (GH) and hepatic GH receptor mRNA [postnatal d (P)28 to P42], insulin-like growth factor (IGF)-I, and IGF-IR (P0 to P42), respectively. Hepatic IGF-I was a major point of control in the GH-IGF axis from P0 to P28. Pituitary GH-releasing hormone receptor may serve an autocrine-paracrine function from P0 to P28, and hypothalamic ghrelin may affect growth by stimulating the release of hepatic IGF-I from embryonic d (E)8 to P28. Hypothalamic ghrelin might interact with corticotropin-releasing hormone (CRH) from P0 to P28. Hepatic IGF-binding protein-2 regulated growth by regulating hepatic IGF-II bioavailability from P0 to P42. Hepatic IGF-binding protein-5 was an important IGF mediator. A coexpression profile was found between hypothalamic GH-releasing hormone (E10 to E16 and P0 to P42), somatostatin (SS; P0 to P28), thyrotropin-releasing hormone (E10 to E16 and P0 to P28), ghrelin (P0 to P42), and pituitary GH mRNA, hypothalamic SS (P0 to P28), corticotropin-releasing hormone (P0 to P42), thyrotropin-releasing hormone (E10 to E18 and P0-P42), and thyroid-stimulating hormone-beta mRNA, respectively. Moreover, AA chickens were fed a nutrient-rich AA diet (as a control group) and LS chickens were fed either a less nutritious LS diet or the AA diet. Langshan and AA chickens fed the same AA diet showed no differences in pituitary GH, hypothalamic SS, ghrelin, hepatic IGF-I, or GH receptor mRNA. Our data indicate that select genes may show parallel expression during certain periods of development, and that differences in BW and gene expression respond differently to nutrient intake in LS and AA chickens. Our findings may help improve the molecular breeding of chickens.

Identification of an (AC)n microsatellite in the Six1 gene promoter and its effect on production traits in Pietrain × Duroc × Landrace × Yorkshire pigs.

The Sine oculis homeobox 1 (Six1) gene is important for skeletal muscle growth and fiber specification; therefore, it is considered as a promising candidate gene that may influence porcine growth and meat quality traits. Nevertheless, the association of Six1 with these processes and the mechanisms regulating its expression remain unclear. The objectives of this study were to identify variant sites of Six1 in different pig breeds, conduct association analysis to evaluate the relationship between polymorphisms of these variants and porcine production traits in Pietrain × Duroc × Landrace × Yorkshire commercial pigs, and explore the potential regulatory mechanisms of Six1 affecting production traits. A total of 12 variants were identified, including 10 single- nucleotide variations (SNVs), 1 insertion- deletion (Indel), and 1 (AC)n microsatellite. Association analysis demonstrated that the SNV, g.1595A>G, was significantly associated with meat color (redness, a*); individuals with the G allele had greater a* values (P < 0.05). Moreover, our results demonstrated that the (AC)n polymorphism in the Six1 promoter was significantly associated with weaning weight (P < 0.05), carcass weight (P < 0.05), and thoracic and lumbar back fat (P < 0.01).In addition, we found that the (AC)n variant was closely related with Six1 expression levels and demonstrated this polymorphism on promoter activity by in vitro experiments. Overall, this study provides novel evidence for elucidating the effects of Six1 on porcine production traits as promising candidate and describes two variants with these traits, which are potential reference markers for pig molecular breeding. In addition, our data on the relationship between porcine Six1 expression and the polymorphic (AC)n microsatellite in its promoter may facilitate similar studies in other species.

Genome-wide association analysis reveals genomic regions on Chromosome 13 affecting litter size and candidate genes for uterine horn length in Erhualian pigs.

Litter size has a great impact on the profit of swine producers. Uterine development is an important determinant of reproduction efficiency and could hence affect litter size. Chinese Erhualian pig is one of the most prolific breeds in the world, even though large phenotypic variation in litter size was observed within Erhualian sows. To dissect the genetic basis of the phenotypic variation, we herein conducted genome-wide association studies for total number born and number born alive (NBA) of Erhualian sows. In total, one significant single nucleotide polymorphism (SNP) (P<1.78e-06) and 11 suggestive SNPs (P<3.57e-05) were identified on 10 chromosomes, confirming seven previously reported quantitative trait loci (QTL) and uncovering six QTL for litter size or uterus length. One locus on Sus scrofa chromosome (SSC) 13 (79.28 to 90.43 Mb) harbored a cluster of suggestive SNPs associated with multiparous NBA. The SNP (rs81447100) within this region was confirmed to be significantly (P<0.05) associated with litter size in Erhualian (n=313), Sutai (n=173) and Yorkshire (n=488) populations. Retinol binding protein 2 and retinol binding protein 1 functionally related to the development of uterus were located in a region of 2 Mb around rs81447100. Moreover, four genes related to embryo implantation and development were also detected around other significant SNPs. Taken together, our findings provide a potential marker (rs81447100) for the genetic improvement of litter size not only in Chinese Erhualian pigs but also in European commercial pig breeds like Yorkshire, and would facilitate the final identification of causative variant(s) underlying the effect of SSC13 QTL on litter size.

Measuring parallel fiber length in the rat cerebellum.

A new approach was devised to obtain the mean length of parallel fibers in Golgi sections of the rat cerebellum. This method was based on the principle that within a given region of the cerebellum the mean length of parallel fibers should be inversely proportional to the likelihood of spotting the ends of parallel fibers. An experimental protocol was designed based on the statistical relationship between the mean fiber length and the number of the ends of parallel fibers for a given total length of parallel fibers examined. Our methodology was simple and could have advantages over other existing methods.

Age-related changes in the cerebellum: parallel fibers.

Between the ages of 3 and 23 months, the cerebella of NIH Fischer 344 rats lose 30% of the thickness of the molecular layer, 60% of the length of parallel fibers, and 80% of the synaptic varicosities along parallel fibers. Nearly 60% of these synaptic varicosities disappear between 3 and 9 months. Thus, the loss of cerebellar synapses is unusual, may have begun early in life, and continue into old age, causing profound synaptic losses in older rats. In addition to serious implications to functional losses, we speculate that these synaptic losses may trigger other age-related cellular losses in the cerebellum.

Electrophysiology of nitrous oxide on cerebellar granule cells: a single-cell study.

We recorded 18 single cells in the granule cell layer of the cat. Each single cell was screened and identified as a granule cell based on a set of criteria derived from known electrophysiological properties of granule cells. We then monitored the effects of nitrous oxide on the spontaneous activities and the auditory responses of these cells. Nitrous oxide consistently caused a severe inhibition of spontaneous activities as well as responses to sound in these cells. Furthermore, the amplitudes of their action potentials decreased during the inhibition. Three of the 18 cells were subsequently injected intracellularly with HRP. All three were verified to be granule cells. In those granule cells we recorded intracellularly, nitrous oxide did not change the resting membrane potentials. The gradual decrease in the amplitudes of action potentials suggested that some of the mechanisms leading to the genesis of action potentials were being altered by nitrous oxide. It is also possible that nitrous oxide may act on synaptic transmission at a site located postsynaptically on the granule cells.

Target areas of presumed auditory projections from lateral and dorsolateral pontine nuclei to posterior cerebellar vermis in rat.

Small injections of the retrograde tracer horseradish peroxidase were made into lobules V through X in the posterior cerebellar vermis of the rat. Labeled cells were counted in the auditory subdivisions of the pontine nuclei, the lateral and dorsolateral pontine nuclei. The results suggest that, within the posterior vermis, lobules VI, VII, and VIII together receive over 90% of projections from the lateral and dorsolateral pontine nuclei.

Nitrous oxide inhibits auditory and visual responses of granule cells in the cerebellum.

Much of the laboratory investigation on the auditory and visual areas of the posterior vermis has been carried out under barbiturate anesthesia. It is now known that barbiturates potentiate GABA inhibition by binding directly to the GABA receptor protein. Since GABAergic receptors are present in many cell types of the cerebellar cortex, barbiturate anesthesia is likely to interfere severely with cerebellar physiology. We have examined auditory and visual responses in granule cells in the cerebellum of the cat under nitrous oxide anesthesia. To our surprise, nitrous oxide abolished auditory as well as visual responses in the granule cell layer in the posterior vermis. However, both auditory and visual responses recovered after the cessation of nitrous oxide.

Thresholdless, hysteresis-free, V-shaped, electro-optical switching for a ferroelectric liquid crystal cell.

A ferroelectric liquid crystal (FLC) cell can be modeled as a combination of capacitors and resistors. In accordance with the properties of the FLC cell, external electric elements, such as capacitors and resistors, are usually connected to achieve a V-shaped performance at a driving inversion frequency fi. However, the inversion frequency is strongly dependent on the external electric elements and the applied voltage. In this paper, the relation between the inversion frequency and the applied voltage is discussed. Additionally, the inversion frequency is found to be approximately proportional to (ReqCeq)(-0.52), where Req and Ceq are equivalent resistance and capacitance, respectively. Based on the above properties, a useful driving scheme is proposed to achieve thresholdless, hysteresis-free, V-shaped characteristics for FLC cells at a driving frequency of 100 Hz. The driving scheme can be applied to fast-response FLC display.

Branchiogenic squamous cell carcinoma: a case report.

Branchiogenic carcinoma is extremely rare. A 41-year-old man presented with a well-defined, fluctuant, painless mass on his right neck and received complete tumour excision. The pathological diagnosis was branchial cleft squamous cell carcinoma, which was characterized microscopically by a branchial cleft cyst with a tumour and a non-tumour transitional zone. In the past, several hundred cases were reported erroneously because of the rather complicated diagnostic criteria. The modified criteria of Khafif introduced in 1989 are more practical, and all were met in this case.

[Phosphorimetric analysis of carbaindoline].

Carbaindoline possesses strong phosphorescence with excitation at 304 mm and emission at 434 nm in water at 77 K. The phosphorescence is stronger in alkaline media than in acid media. Kl enhances the phosphorescence of carbaindoline but shortens the life time. The heavy cations Cu2+, Fe3+, Pb2+, Tl3+, Zn2+, and molybdate do not enhance the phosphorescence, but some of them, Cu2+ and Fe3+, have quenching effect on carbaindoline phosphorescence. The phosphorescence of nine carbaindoline analogues was studied. After oxidation of > CH2 at position 2 to > C = O, the phosphorescence decreased significantly. The phosphorimetric method for analysis of carbaindoline was established with a detection limit of 5 ng/ml. Ten minutes after ip administration of carbaindoline 0.1 mg/kg to mice, the blood drug concentration was found to be 149 +/- 27 ng/ml.

[Studies on biotransformation of jing song ling, a new anesthetic].

Jing song ling (2,4-xylidinothiazoline, XT) is a new anesthetic agent for veterinary uses in China. From urine of rats given the drug three metabolites (MA1, MA2 and MB3) were isolated with structure shown in page 98 in addition to the parent compound (MB1). Preliminary observations showed that the toxicity and pharmacological activity of MB3, MA2 and MA1 decreased significantly compared with the parent drug XT. Species difference of biotransformation of XT between rat and mouse was also observed.

[The biotransformation of cui xing ning in the isolated perfused rat liver].

Similar to eserine, Cui Xing Ning [CXN),5-(1,3,3-trimethylindoline) N, N-dimethyl carbamate] is a choline esterase (ChE) inhibitor. The biotransformation of CXN, which had been studied in vivo and in vitro unsuccessfully, was studied by using isolated perfused rat liver (IPL) method. Six metabolites of CXN from the perfusate were separated by HPLC, and identified with MS, 1HNMR or compared with the authentic compounds. Metabolite I is unchanged CXN, the other are oxidized metabolites. Most of the metabolites lost their activities of ChE inhibition strikingly except metabolite III which retained the 2-position unchanged. The toxicities of some metabolites (II,V) decreased obviously.