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The worldwide chicken gene pool encompasses a remarkable, but shrinking, number of divergently selected breeds of diverse origin. This study was a large-scale genome-wide analysis of the landscape of the complex molecular architecture, genetic variability, and detailed structure among 49 populations. These populations represent a significant sample of the world's chicken breeds from Europe (Russia, Czech Republic, France, Spain, UK, etc.), Asia (China), North America (USA), and Oceania (Australia). Based on the results of breed genotyping using the Illumina 60K single nucleotide polymorphism (SNP) chip, a bioinformatic analysis was carried out. This included the calculation of heterozygosity/homozygosity statistics, inbreeding coefficients, and effective population size. It also included assessment of linkage disequilibrium and construction of phylogenetic trees. Using multidimensional scaling, principal component analysis, and ADMIXTURE-assisted global ancestry analysis, we explored the genetic structure of populations and subpopulations in each breed. An overall 49-population phylogeny analysis was also performed, and a refined evolutionary model of chicken breed formation was proposed, which included egg, meat, dual-purpose types, and ambiguous breeds. Such a large-scale survey of genetic resources in poultry farming using modern genomic methods is of great interest both from the viewpoint of a general understanding of the genetics of the domestic chicken and for the further development of genomic technologies and approaches in poultry breeding. In general, whole genome SNP genotyping of promising chicken breeds from the worldwide gene pool will promote the further development of modern genomic science as applied to poultry.

Endometrial cancer is the most common gynecological malignancy, affecting up to 3% of women at some point during their lifetime (Morice et al., 2016; Li and Wang, 2021). Based on the pathogenesis and biological behavioral characteristics, endometrial cancer can be divided into estrogen-dependent (I) and non-estrogen-dependent (II) types (Ulrich, 2011). Type I accounts for approximately 80% of cases, of which the majority are endometrioid carcinomas, and the remaining are mucinous adenocarcinomas (Setiawan et al., 2013). It is generally recognized that long-term stimulation by high estrogen levels with the lack of progesterone antagonism is the most important risk factor; meanwhile, there is no definite conclusion on the specific pathogenesis. The incidence of endometrial cancer has been on the rise during the past two decades (Constantine et al., 2019; Gao et al., 2022; Luo et al., 2022). Moreover, the development of assisted reproductive technology and antiprogestin therapy following breast cancer surgery has elevated the risk of developing type I endometrial cancer to a certain extent (Vassard et al., 2019). Therefore, investigating the influence of estrogen in type I endometrial cancer may provide novel concepts for risk assessment and adjuvant therapy, and at the same time, provide a basis for research on new drugs to treat endometrial cancer.

Extracellular traps released by neutrophils (neutrophil extracellular traps, NETs) are a double-edged sword, and understanding the mechanism of NET formation is of great significance for disease treatment. However, the short lifespan, the large individual differences, and the inability to perform gene editing render it difficult to decipher NET formation using neutrophils. It is necessary to find a model cell to replace neutrophils to study the mechanism of NET formation. In this study, we used different concentrations (0, 0.1, 1, and 10 μmol/L) of all-trans retinoic acid (ATRA) to differentiate HL-60 cells for different days (1, 3, 5, and 7 days). By detecting the cell viability and nuclear morphology of cells, we confirmed that HL-60 cells were differentiated to neutrophil-like cells (dHL-60) after treated with ATRA for at least 5 days. Using immunofluorescence staining to detect the formation of NETs, we demonstrated that dHL-60 cells differentiated for 5 days with 1 μmol/L ATRA could generate NETs comparable to those produced by neutrophils upon phorbol 12-myristate 13-acetate (PMA) stimulation, without histone H3 citrullination. Furthermore, the formation of NETs by dHL-60 cells were NADPH-dependent and PAD4-independent, consistent with neutrophils. Taken together, these observations suggest that dHL-60 cells differentiated with 1 μmol/L ATRA for 5 days can be used as a model cell for neutrophils to study the mechanism of NET formation.

The radial force of the degradable esophageal stent before and after degradation is one of the important indicators for effective treatment of esophageal stricture. Based on a combination of in vitro experiments and finite element analysis, this paper studies and verifies the biomechanical properties of a new type of degradable esophageal stent under different esophageal stricture conditions. Under radial extrusion conditions, the maximum stress at the port of the stent is 65.25 MPa, and the maximum strain is 1.98%; The peak values of stress and strain under local extrusion and plane extrusion conditions both appear in the extrusion area and the compression expansion area at both ends, which are respectively 48.68 MPa, 46.40 MPa, 0.49%, 1.13%. The maximum radial force of the undegraded stent was 11.22 N, and 97% and 51% of the maximum radial force were maintained after 3 months and 6 months of degradation, respectively. The research results verify the safety and effectiveness of the radial force of the new degradable esophageal stent, and provide a theoretical basis for the clinical treatment of esophageal stricture.

There are five-year transitional period for manufacturers after the issue of regulation(EU) 2017/746 on in vitro diagnostic medical devices(IVDR) in May, 2017. The article introduces the background, major change, the impact of new regulation and the coping strategies of enterprises, the main preparation work should be taken etc, to provide reference or help for the domestic manufacturers when they prepare the CE marking according to IVDR.

Endophytic fungus is an important treasure trove for discovery of structurally unusual and biologically diverse compounds. A phytochemical investigation on a fungus Clonostachys rosea inhabits inner tissue of Blumea balsamifera (L.) DC. was initiatedrecently in our lab. Six pure compounds were isolated through silica gel column chromatography, sephadex LH-20, and semi-preparative HPLC techniques, with bio-guided strategy. Their structures were characterized as verticillin A (1), (S)-(+)-fusarinolic acid (2), 8-hydroxyfusaric acid (3), cerebroside C (4), 3-Maleimide-5-oxime (5), and bionectriol A (6) by analyses of NMR and MS data. All compounds were tested in vitro antibacterial activities against four strains of bacteria, Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Pseudomonas aeruginosa, and results revealed that 1, 4 and 6 display notableinhibition againstthree bacteria, with MIC values ranging from 2 to 16 μg/mL. Our findings provide references for mining novel antibiotics from endophytes originated from Li Minority medicinal plant B. balsamifera (L.) DC.

Seamless modification is a popular genomic manipulation technique in genetic engineering. Selection stringency of the counter-selection system determines the efficiency of the seamless modification. Recently, a novel counter-selection system, kil, was constructed. It is reported that the selection selectivity of kil is higher in host bacteria harboring plasmid pSim6 than that harboring pKD46, indicating that recombinants could be selected out more efficiently by combining kil counter-selection system and plasmid pSim6. In order to confirm this speculation, four different loci (lacI, dbpa, ack, glk) in Escherichia coli strains W3110, MG1655 and DH10B were selected for testing: dsDNA fragments of different sizes (500 bp, 1 000 bp, and 2 000 bp) were used to substitute tet/kil. As expected, recombination efficiency was higher in host bacteria harboring plasmid pSim6 than that harboring pKD46, and the results were more obvious with the length of dsDNA increasing. Specifically, recombination efficiency was 1.2 to 2 fold higher in pSim6 harboring bacteria than in pKD46 harboring bacteria when dsDNA fragments were 1 000 bp in length. With the length of dsDNA increasing up to 2 000 bp, the gap increased to 2.2-5 fold. In conclusion, it is easier to perform seamless modification by combining kil counter-selection system and plasmid pSim6 than combining kil and pKD46. An alternative tool in genomic engineering is provided in this study.

Objective: To study the effect of aldosterone on cell proliferation, alkaline phosphatase (AKP) activity and osteogenic gene expression in rat osteoblasts and explore the mechanisms., Methods: Osteoblasts isolated from the skull of neonatal SD rats by enzyme digestion were cultured and treated with different concentrations of aldosterone. The cell proliferation and AKP activity were evaluated using CCK-8 assay kit and AKP assay kit, respectively. The effects of aldosterone on mRNA and protein expressions of the osteogenic genes and epithelial sodium channel (ENaC) gene were investigated using semi-quantitative PCR and Western blotting., Results: Compared with the control cells, the cells treated with 0.01-1.0 µmol/L aldosterone showed obviously enhanced proliferation while lower (1×10 -3 µmol/L) or higher (10 µmol/L) concentrations of aldosterone did not significantly affect the cell proliferation. Aldosterone within the concentration range of 1×10 -3 to 10 µmol/L did not cause significant changes in AKP activity in the osteoblasts. Treatment with 0.01 to 1.0 µmol/L aldosterone significantly upregulated the expressions of the osteogenic genes and α-ENaC gene at both the mRNA and protein levels., Conclusion: Aldosterone within the concentration range of 0.01-1.0 µmol/L stimulates the proliferation and osteogenic gene expressions and enhances α-ENaC gene expression in rat osteoblasts in vitro, suggesting the possibility that ENaC participates in aldosterone-mediated regulation of osteoblast functions.

Objective To design, construct and express a chimeric αβ TCR harboring the immunoglobulin-like (Ig) domain of γδ TCR in Jurkat T cells. Methods The fusion sites of TCR δIg were determined by bioinformatics analysis. Then the protein structures of TCR α δIg and TCR β δIg were predicted by homology modeling. Furthermore, the structures of TCR α δIg and TCR β δIg were compared with the wild type (wt) TCR α and TCR β respectively by combinatorial extension (CE). After that, the TCR α δIg and TCR β δIg were fused to fluorescent protein ECFP and EYFP respectively via the overlap PCR, and then the fusion genes (TCR α δIg-ECFP and TCR β δIg-EYFP) were cloned into pIRES2-EGFP vector and respectively located at the upstream and downstream of an internal ribosome entry site (IRES). The recombinant prokaryotic expression vector pIRES-TCR βδIg-EYFP/TCR αδIg-ECFP was transferred into Jurkat T cells. Finally, the expression of TCR δIg in Jurkat T cells was monitored by confocal laser scanning microscopy (CLSM). Results The variable region structure of the TCR δIg did not change and the antigen recognition active regions remained stable compared to the wtTCR. The recombinant expression plasmid was successfully constructed as confirmed by PCR identification and sequencing analysis. CLSM showed that TCR δIg was expressed and located at the plasma membrane of Jurkat T cells. Conclusion The design of TCR δIg was reasonable and the TCR δIg could be expressed on Jurkat T cell surface.

Aim: To prepare CD80-scFv in CHO cells and investigate its effect on the recognition and proliferation of different tumor cells., Methods: The CD80-scFv was purified from culture supernatant without fetal calf serum (FCS) by IMAC affinity chromatography, and then bound to CD80 molecule on the Daudi, U251, A375 and 8266 cells detected by flow cytometry. Different concentrations of CD80-scFv were added in the training system of different tumor cells, and we analyzed the influence on the proliferation of these cells by MTT assay. With 8266 cells which expressed CD80 highly and were treated by mitomycin firstly as the APC, and human PBMC as the effect cells, we analyzed the influence of the CD80-scFv on the proliferation of PBMC by blocking the co-stimulatory signal., Results: The concentration of CD80-scFv purified from FCS was about 6.67 mg/L and the binding rate of CD80-scFv with Daudi, U251, A375, 8266 and U266 were 96.8%, 39.6%, 20.5%, 99.9% and 3.8%, respectively. CD80-scFv suppressed the proliferation of Daudi and 8266 cells which expressed CD80 highly, and the inhibition rate of Daudi and 8266 cells cultured with CD80-scFv (the final concentration was 25 μg/mL) was 24.04% and 24.16%, respectively. Additionally, the antibody suppressed the proliferation of PBMC by blocking the co-stimulatory signal mediated by CD80-CD28., Conclusion: CD80-scFv can recognize CD80 molecule and suppress the proliferation of tumor cells which express CD80 highly.

To efficiently use both cellulose and hemicellulose for lipid production, rice straw was hydrolyzed by a two-step process including dilute acid pretreatment and then enzymatic hydrolysis, and the hydrolysate was used as carbon source for lipid fermentation by Trichosporon fermentans HWZ004. After a simple overliming, the concentrations of acetic acid, furfural and 5-hydroxymethyl-furfural were 0.4 g/L, 0.1 g/L and 0.05 g/L, respectively. The hydrolysate could be used for lipid fermentation with T. fermentans HWZ004 without adding other nutrients except for a small amount of nitrogen source and trace CuSO4.5H2O. The optimum inoculum size, initial pH and temperature were 5.0%, 7.0 and 25 degrees C, respectively. A total biomass of 26.4 g/L with a lipid content of 52.2% (corresponding to a lipid yield of 13.8 g/L) was achieved after cultivation of T. fermentans HWZ004 under the above-mentioned conditions for 7 days. The lipid coefficient (lipid yield on substrate consumed) is 17.0, which is much higher than the corresponding one (11.9) obtained on detoxified rice straw hemicullulose hydrolysate by original yeast T. fermentans CICC 1368. The fatty acid composition of the lipid was found to be similar to that of vegetable oil and its unsaturated fatty acid content was over 70%, thus the lipid is a promising material for biodiesel production.

Aim: To investigate the roles of autocrine soluble FN of MG-63 cells in cell shape maintaining., Methods: Human osteosarcoma MG-63 cells (1 × 10(4); cells/cm(2);) were routinely cultured in MEM medium+10% fetal bovine serum (FBS) for 24 h and then the medium was replaced by fresh medium(the group without autocrine soluble FN), conditional medium of collected after 24 h of MG-63 cells (the group containing autocrine soluble FN ) and fresh medium with 20 microg/mL FN( the group with plasma FN) after 24 h, and the cell morphology was observed using phase contrast microscope; The concentrations of soluble FN in fresh and conditional medium were detected; Microfilament and insoluble FN changes of MG-63 cells were investigated., Results: The concentration of soluble FN in conditional medium is much higher than that in the fresh medium (P<0.01). Fresh medium will result in the cell shape changing from spindle to round for 0.5 h. The cell began to spread for 1 h, and the cell shape recovered for 2 h; Conditioned medium has no significant effects on cell shape. Addition of FN (20 mg/L) to fresh medium could inhibit the cell shape change induced by fresh medium. Microfilament and insoluble FN were disorganized by fresh medium., Conclusion: These results indicated that autocrine soluble FN of osteosarcoma MG-63 cells may be involved in cell shape maintaining.

A survey indicated that the prevalence of Demodex infection among 512 college students in Tangshan was 36.3% (186/512), that of males and females was 39.3% (81/206) and 34.3% (105/306) respectively (P>0.05). The infection of Demodex folliculorum accounted for 82.3% (153/186), followed by D. brevis (7.5%, 14/186) and mixed infection (10.2%, 19/186). The prevalence was 47.0% (93/198) in subjects with oily skin, 26.6% (37/139) in those with dry skin, and 33.9% (56/165) in mixed-type skin (P<0.05). Subjects with facial diseases (62.0%, 75/121), such as rosacea and acne, were more likely to be infected with Demodex than those with healthy skin (27.6%, 80/290) (P<0.05). Prevalence in those lived in humid environment (67.9%, 95/140) was higher than those lived in the desiccating environment (24.5%, 91/372) (P<0.05).

Objective: To investigate the prevalence of Entamoeba gingivalis infection in college students in Tangshan, and analyze the relationship between the infection and human behaviors., Methods: 551 students of grades 1-3 from six colleges in Tangshan received questionnairing, which covered the oral health state, teeth-brushing, xylitol gum-chewing, diet fondness, and smoking. Specimens were taken from the tooth surface of the lesion or fouling materials by using disinfected toothpicks and the smears were observed microscopically to examine Entamoeba gingivalis infection., Results: The prevalence of Entamoeba gingivalis infection was 28.3% (156/551), 30.4% (55/181) and 24.6% in males and females (91/370) respectively (chi2=2.09, P>0.05). The prevalence in students with or without oral disorders was 41.2% (84/204) and 20.8% (72/347) respectively, with a significant statistical difference (chi2=26.41, P<0.01); it was 22.5% (53/236) and 32.7% (103/315) among students who cleaned their teeth regularly or irregularly (chi2=6.97, P<0.01); it was 18.3% (17/93) and 30.4% (139/458) among those usually with or without chewing xylitol gum (chi2=5.55, P<0.05)., Conclusion: Entamoeba gingivalis infection is common in the college students in Tangshan and it has a close relation to the oral hygiene habits and the presence of oral disorders.

The full length cDNA of silkworm hibadh gene was cloned by RT-PCR and RACE (Rapid amplification of cDNA ends) technique. The hibadh gene and its deduced amino acid sequences were analyzed. The tissue distribution of hibadh gene in 5th instar silkworm larvae was tested by RT-PCR. The expression patterns of hibadh gene in simulated weightless environment were analyzed by real time RT-PCR. The results showed that the full length hibadh cDNA sequence was 1074 bp in lenth, including an open read frame of 969 bp encoding the entire coding region of Hibadh (GenBank accession No. EU719652). The deduced amino acid sequence similarities of hibadh between silkworm and Burkholderia ambifaria, Drosophila melanogaster, Apis mellifera, Xenopus tropicalis, Mus musculus, Homo sapiens were 46%, 43%, 48%, 44%, 45%, 45%, respectively. Signal peptide analysis showed that Hibadh was a secretory protein. There wasn't glycosyl-phosphatidyl inositol anchor site in Hibadh amino acid sequence. Molecular weight and isoelectric point of Hibadh were 34.1 kD and 9.14 respectively. The RT-PCR tests indicated that the hibadh gene expressed in head, silk gland, midgut, cuticle, blood, fat body, tuba malpighii of the 5th instar silkworm larvae. There were different expression patterns of hibadh gene during different silkworm embryo period in simulated weightless environment. Simulated weightlessness resulted in the expression of silkworm hibadh gene up regulated 2.3-fold (P < 0.05), up regulated 4.6-fold (P<0.01), down regulated 7.6-fold (P < 0.01), down regulated 2.6-fold (P < 0.05) during apophysis formation period, inverse period, trachea formation period, and whole embryo period, respectively. There was no significant change of hibadh gene expression during other period of silkworm embryo between simulated weightless and control groups. There were different response patterns to simulated weightless environment between hibadh gene and whole body of silkworm. Gene showed much higher sensitivity compared to whole body in response to environment. This study is useful for the further research on the gravity biological mechanism of hibadh gene.

Aim: To obtain sufficient numbers of pure and activated human primary NK cells for potential clinical application., Methods: PBMC based ex vivo expansion of natural killer cells was set up. The expansion was initiated by co-culture of PBMC and stimulator cells (irradiate genetic modified K562 cells) in RPMI1640 medium with 1 000 U/mL IL-2. Genetic modified K562 cells 'K562D3' were prepared by expressing IL-15, 4-1BBL and IL-18 on K562 cell membrane., Results: An average of 500 fold expansion of CD56(+)CD3(-) cells was observed after 3 weeks of co-culture. The NK cells population could reach 93% after expansion, comparing with 7% before expansion. The expanded NK cells lysed 95% of K562 targets in a 5:1 effector to target ratio. IL-15/4-1BBL bound K562 stimulatory cells could expand same fold NK cells as K562D3, but the cytotoxicity of NK cells expanded by 'K562D3' was 10% higher., Conclusion: The described method is a simple and efficient way for the expansion of human NK cells.

Cold-induced genes of highland barley (Hordeum vulgare L. var. nudum Hk. f.) were studied using suppression subtractive hybridization (SSH) technique. The cDNA from the materials treated with 4 degrees C was used as "tester", and that from the materials growing in green house (20+/-2 degrees C) as "driver". A subtractive library of highland barley including 640 cDNA clones was constructed in this study. Enzyme digestion of 32 clones chosen randomly from the library indicated that 87.5% of them contained inserts. The cDNA inserts of 16 clones were sequenced. Blast search analyses showed that these cDNAs were homologies to genes encoding the following proteins: metallothionein, protein kinase, ethylene signal transcription factor, bZIP transcription factor, zing finger transcription factor, ribulose-1,5-bisphosphate carboxylase, ribosomal protein, sodium: hydrogen antiporter, catalase, NADPH-cytochrome reductase, ascorbate peroxidase, DNA binding protein, and sugar transporter-like protein. These results indicated that the cDNA clones in the library were related to cold-induced genes, and suggested that the cold-tolerant mechanism of highland barley might be a complicated, interactive system involving multiple approaches and genes. Construction of subtractive cDNA library provided an advantage for further studies to isolate and clone cold-induced genes in highland barley.

The actin-depolymerizing factor 4 gene (ADF4) of Arabidopsis thaliana was cloned and sequenced (Figs.1, 2). The plant expression vector with ADF4 was constructed and transformed into tobacco by Agrobacterium tumefaciens. Molecular identification showed that the ADF4 gene was integrated into the genome of tobacco and expressed in transgenic tobaccos assayed by PCR and RT-PCR (Fig.3). Expression of an Arabidopsis thaliana ADF4 gene in tobacco resulted in morphological change of plants. The effects of ADF4 on transgenic tobaccos growth were as follows: the hypocotyls of transgenic plants were wavy, especially in darkness, whereas those of the control were straight (Fig.4A); the root hairs of transgenic plants were less than the control, and they were also wavy (Fig.4B); the parenchyma cells of transgenic plants were larger than the control and the arrangement of vascular bundle was out of order (Fig.4C); the flowering time of T(2) line was at least 7 days later than the control; the pollen tubes of transgenic plants were shorter than those of the control (Fig.4D).

Aim: To investigate the response of hypothalamic corticotropin releasing factor (CRF) and arginine vasopressin (AVP) of adrenalectomized neonatal rats to hypoxia., Methods: The hypoxia was simulated in hypobaric cabin. The amount of arginine vasopressin (AVP) and corticotropin releasing factor (CRF) was assayed by RIA method., Results: When neonatal rats exposed to acute hypoxia (simulated altitude of 5 000 m and 7 000 m, 24 h), their hypothalamic CRF was not changed in 3 d and 7 d rats, lower than control in 14 d, 21 d and 28 d rats, while hypothalamic AVP had no change in 3 d rats, was lower than control in 14 d and higher in 7 d, 21 d and 28 d rats. The responsive pattern of those two neuropeptides to acute hypoxia changed with the growth of rats. Adrenalectomy reduced the amount of hypothalamic CRF and AVP in 14 d, 21 d and 28 d rats, except AVP in 28 d rats. When adrenalectomized neonates exposed to hypoxia, there was no further change in CRF and AVE, means adrenalectomy not only removed the negative regulation of corticosteroid on CRF and AVE, but also inhibited the normal development of CRF and AVP neurons and thus, their responsive ability to hypoxia.

Aim: To investigate the influence of hypoxia on postnatal developing pattern of hypothalamic corticotropin releasing factor(CRF) and arginine vasopressin (AVP) in male and female neonatal rats., Methods: The hypoxia was simulated in hypobaric cabin. The content of AVP and CRF was assayed by RIA method., Results: It was showed the female rats had the similar developing process with male rats, either at 2300 m or at 5000 m altitude. When they developed at simulated 5000 m altitude, the content of their hypothalamic CRF was lower than that of control rats on postnatal day 21, while the content of AVP was higher than control on day 21 and 28., Conclusion: Maybe the difference of the function between hypothalamic CRF and AVP or/and the incoherence of their developing stage contribute to their particular developing pattern at 5000 m altitude.

A total of 354 isolates of actinomycetes, of which 76 were detected cytotoxic activity was isolated from near-shore marine samples collected at Wenchang mangrove, DanZhou harbor and YanPu harbor. Four isolation methods were employed, which are SDS pretreatment, phenol pretreatment, heating pretreatment and potassium dichromate selection culture, and media such as'Yeast extract-Malt extract (YE), Glucose-Asprine (GA), Starch-Casin (SC), Starch-KNO3 (Gause) were used. It was showed that heating pretreatment and potassium dichromate selection culture were more considerable methods for extensive isolation of actinomycetes. Medium YE and Gause showed best results in both the total number of actinomycetes and the number of active isolates against tumor cell B16. The genotypic diversity of 30 strains of Streptomycetes possessing strong cytotoxic activity against B16 cell (ID50 > or =200) was analyzed by 16S ARDRA, which resulted in 17 RFLP types, and indicated relatively rich genotypic diversity among these Streptomycetes. 16S rDNA sequence analysis of three strains, 050642, 060386 and 060524 (ID50 > or = 1200) further confirmed that they all belong to Streptomyces genus and strain 050642 was suggested a novel Streptomyces. Spp with the highest similarity of 95% to Streptomyces cattleya.