ObjectiveCandidate genes related to the regulation of litter size traits in English Springer Spaniel breeding bitches are explored, and the genetic mechanism underlying fertility in this breed is investigated, in order to provide reference molecular markers for genomic selection of high fecundity. MethodsWhole genome resequencing was performed on English Springer Spaniel breeding bitches that had given birth to at least 3 litters, and the bitches were divided into a high-litter-size group and a low-litter-size group according to the average litter size. Selection signal analysis was used to obtain the intersection of fixation index (F_st) and nucleotide diversity (Pi) signals as highly selected regions, and candidate genes were screened based on gene annotation and functional enrichment analysis. ResultsThe average litter size in the high-litter-size group (7.41±1.27) was significantly higher than that in the low-litter-size group (3.82±1.20) (P＜0.05), and the total number of live offspring in the high-litter-size group (7.06±1.10) was extremely significantly higher than that in the low-litter-size group (3.67±1.11) (P＜0.01). A total of 3 155 706 SNPs were detected in the two groups, 63.09% of which were located in intergenic regions, 33.96% in intronic regions, and 0.38％, 0.57％, and 0.09％ in exonic regions, 3' untranslated regions (UTRs), and 5'UTRs, respectively. Among the SNPs in exonic regions, 5 256 were nonsynonymous variants, accounting for 43.55%. A total of 1 752 differential genes were identified after annotation screening. Gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG) enrichment analyses identified 13 candidate genes that may affect reproductive performance and litter size traits, including WDR35, SMAD7, RPGR, RERGL, PGRMC2, LOC482182, GIMD1, COX7B2, COX16, BMPR2, BMP6, BICD1, and SLC9C1. Their functions mainly involve reproductive hormone regulation, embryonic development, GTPase activation, and oocyte apoptosis. ConclusionEnglish Springer Spaniel breeding bitches have undergone significant artificial selection for litter size traits. These 13 candidate genes play key roles in oocyte maturation and regulation during early pregnancy, providing a new molecular basis for elucidating the genetic mechanism of canine reproductive traits.

AIM:To explore the role of miR-195-5p in mediating trastuzumab resistance in gastric cancer and to validate its potential as a therapeutic target along with its target gene GPRC5A.METH-ODS:Trastuzumab-resistant gastric cancer cell lines(NCI-N87 and MKN45)were established.Cell viabili-ty under trastuzumab treatment was assessed us-ing CCK-8 assays.Expression levels of miR-195-5p were determined by RT-qPCR.Transfection with miR-195-5p mimics was performed to evaluate changes in trastuzumab sensitivity and prolifera-tion.GPRC5A expression was also measured by RT-qPCR,and the targeting relationship between miR-195-5p and GPRC5A was confirmed using a dual-lu-ciferase reporter assay.RESULTS:Parental cells showed higher sensitivity to trastuzumab than re-sistant cells,with miR-195-5p expression signifi-cantly lower in the latter.Overexpression of miR-195-5p in resistant cells enhanced trastuzumab sen-sitivity and reduced proliferation.GPRC5A was found to be upregulated in resistant cells,and miR-195-5p directly targeted GPRC5A,affecting cell pro-liferation under trastuzumab treatment.CONCLU-SION:miR-195-5p may regulate trastuzumab sensi-tivity in gastric cancer by targeting GPRC5A,sug-gesting potential as a molecular marker for trastu-zumab therapy guidance.

AIM:To explore the role of miR-195-5p in mediating trastuzumab resistance in gastric cancer and to validate its potential as a therapeutic target along with its target gene GPRC5A.METH-ODS:Trastuzumab-resistant gastric cancer cell lines(NCI-N87 and MKN45)were established.Cell viabili-ty under trastuzumab treatment was assessed us-ing CCK-8 assays.Expression levels of miR-195-5p were determined by RT-qPCR.Transfection with miR-195-5p mimics was performed to evaluate changes in trastuzumab sensitivity and prolifera-tion.GPRC5A expression was also measured by RT-qPCR,and the targeting relationship between miR-195-5p and GPRC5A was confirmed using a dual-lu-ciferase reporter assay.RESULTS:Parental cells showed higher sensitivity to trastuzumab than re-sistant cells,with miR-195-5p expression signifi-cantly lower in the latter.Overexpression of miR-195-5p in resistant cells enhanced trastuzumab sen-sitivity and reduced proliferation.GPRC5A was found to be upregulated in resistant cells,and miR-195-5p directly targeted GPRC5A,affecting cell pro-liferation under trastuzumab treatment.CONCLU-SION:miR-195-5p may regulate trastuzumab sensi-tivity in gastric cancer by targeting GPRC5A,sug-gesting potential as a molecular marker for trastu-zumab therapy guidance.

Heart failure is the leading cause of death worldwide. Compound Danshen Dripping Pill (CDDP) or CDDP combined with simvastatin has been widely used to treat patients with myocardial infarction and other cardiovascular diseases in China. However, the effect of CDDP on hypercholesterolemia/atherosclerosis-induced heart failure is unknown. We constructed a new model of heart failure induced by hypercholesterolemia/atherosclerosis in apolipoprotein E (ApoE) and LDL receptor (LDLR) dual deficient (ApoE^-/-LDLR^-/-) mice and investigated the effect of CDDP or CDDP plus a low dose of simvastatin on the heart failure. CDDP or CDDP plus a low dose of simvastatin inhibited heart injury by multiple actions including anti-myocardial dysfunction and anti-fibrosis. Mechanistically, both Wnt and lysine-specific demethylase 4A (KDM4A) pathways were significantly activated in mice with heart injury. Conversely, CDDP or CDDP plus a low dose of simvastatin inhibited Wnt pathway by markedly up-regulating expression of Wnt inhibitors. While the anti-inflammation and anti-oxidative stress by CDDP were achieved by inhibiting KDM4A expression and activity. In addition, CDDP attenuated simvastatin-induced myolysis in skeletal muscle. Taken together, our study suggests that CDDP or CDDP plus a low dose of simvastatin can be an effective therapy to reduce hypercholesterolemia/atherosclerosis-induced heart failure.

Adaptation, Physiological ; Adenosine Monophosphate ; administration & dosage ; analogs & derivatives ; pharmacology ; therapeutic use ; Administration, Intranasal ; Alanine ; administration & dosage ; analogs & derivatives ; pharmacology ; therapeutic use ; Animals ; Betacoronavirus ; genetics ; physiology ; Chlorocebus aethiops ; Coronavirus Infections ; drug therapy ; virology ; Disease Models, Animal ; Female ; Host Specificity ; genetics ; Lung ; pathology ; virology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mutation, Missense ; Nasal Mucosa ; virology ; Pandemics ; Pneumonia, Viral ; drug therapy ; virology ; RNA, Viral ; administration & dosage ; genetics ; Turbinates ; virology ; Vero Cells ; Viral Load ; Virus Replication

This paper aimed at evaluating the therapeutic effects of acupuncture and moxibustion in treatment of heart-spleen deficiency and observing the improvement effect for insomnia patients′ sleep quality. A total of 70 cases of heart-spleen deficiency were selected as the research data, to the all cases were divided into acupuncture group and controlgroup. Acupuncture group were treated with acupuncture and moxibustion in many acupoints, the control group weretreated with estazolampo. After 3 courses of treatment, the PSQI and TESS of the two groups were observed. The totaleffective rate of the acupuncture group was 94.3％, the total effective rate of the control group was 77.1％ (P ＜ 0.05), andthe acupuncture group was more ideal than the control group (P ＜ 0.05) . After treatment the PSQI score and TESS scorein acupuncture group were decreased compared with control group (P ＜ 0.05) . Acupuncture and moxibustion in thetreatment of heart-spleen deficiency insomnia curative effect is ideal and reliable, can effectively decrease the PSQI andTESS score, and has lower auxiliary work compared with taken estazolam.

The patients with mechanical prosthetic valve replacement need anticoagulant therapy for all their life. The incidence of thromboembolism and anticoagulation-related bleeding events still account for major postoperative complications after mechanical heart valve replacement. Most of the complications happen in the first half year after operation. Therefore, early anticoagulation therapy is very important. Of course, so far most guidelines focus stating their opinions on long-term anticoagulant therapy. However, there is no consensus about anticoagulant therapy in the early period of postoperation. In this review, we summarize early anticoagulant therapy of the patients with mechanical heart valve replacement through consulting domestic and abroad relevant research in recent years and give an overview of the present situations of early anticoagulant therapy.

Objective To illustrate that tumor necrosis factor-α (TNF-αt) / nuclear factor of activated T cells pathway is the main function way that mesenchymal stem cells (MSC) inhibit proliferation of pulmonary vascular smooth muscle cells during treating pulmonary hypertension (PH).Methods MSC from human umbilical cord was used to treat PH rat models induced by monocrotaline.Rats were divided into the control group,the PH model group and the MSC group.The general conditions of the rats were observed.Haemodynamics was detected.Pathological sections and immunohistochemistry method were used to detect the lung structure and tissue changes.Changing conditions of TNF-αt/NFAT were detected.Results Compared with rats in the PH model group,the general conditions of the MSC group tended to be normal evidently:the right ventricular systolic pressure (RVSP) dropped [(30.37 ±3.13) mmHg vs.(47.90 ± 3.45) mmHg,1 mmHg =0.133 kPa],the aortic pressure (MAoP) increased [(115.03 ± 16.01) mmHg vs.(92.78 ± 16.28 mmHg)],the thickening condition of arterial intima-media was evidently relieved [(17.22 ±1.21)％ vs.(31.68 ±2.26)％],the plasma TNF-α level decreased obviously [(842 ±76) ng/L vs.(245 ±24)ng/L],and the lung tissue TNF-o level decreased (0.172 ±0.024 vs.0.248 ± 0.051),and all the differences were statistically significant (all P ＜ 0.05).The activation of pulmonary artery NFATc2 in the MSC treatment group was apparently inhibited.Conclusions MSC therapy may perform the treating effect in PH by inhibiting the over-proliferation of inflammation related pulmonary vascular smooth muscle cells via TNF-oα/NFAT pathway.

Objective Lamins are the major components of nuclear lamina underneath the inner nuclear membrane (INM).Lamins express in most cells and are involved in the whole process of growth, also play a major role in cell stability and embryonic development.Mutant in human LMNA gene may lead to a series of disorders, which are similar to progeria or other aging-associate syndrome.In this study, we report a new lmna knockdown animal model generated in our laboratory in order to provide a useful tool for studying laminopathies.Methods Two plasmids tagged to zebrafish lmna gene were designed based on morpholino oligonucleotides technology.Co-microinjected the plasmids into zebrafish embryos to knockdown lmna gene.Imagining and western blot detection were used to identify the mutants.Results Two different proteins, Lamin A/C, were expressed in the zebrafish embryos.Two plasmids lmna-MO and lmna-EGFP-pCS 2 + were generated and co-microinjected into embryos.The results of imagining and western blot showed that the expression of lmna gene was downregulated in the zebrafish embryos.Conclusions Lamin A/C are expressed in zebrafish.lmna gene can be knocked down by the injection of lmna-MO and lmna-EGFP-pCS 2 +.This new animal model may be a powerful tool for study on laminopathies.

Objective:To observe the clinical effect of acupuncture therapy to support yang and consolidate the constitution for cervical spondylosis of vertebral artery type (CSA). Methods:A total of 72 patients with CSA were randomly divided upon the random digital table into an acupuncture- moxibustion group and an acupuncture group, 36 cases in each group. The acupuncture-moxibustion group was treated by electroacupuncture (EA) at bilateral Fengchi (GB 20) and C4-6 Jiaji (EX-B 2), in combination of moxibustion at Baihui (GV 20), Shenshu (BL 23) and Mingmen (GV 4) to support yang and consolidate constitution. The acupuncture group was given by routine EA at bilateral Fengchi (GB 20), cervical Jiaji (EX-B 2), Tianzhu (BL 10), Jianjing (GB 21), Houxi (SI 3), Hegu (LI 4) and Waiguan (TE 5). The treatment was given once every day and continuous six sessions made one course. After 2-course treatment, the improved situation of clinical symptoms and clinical effect were observed in the two groups. Results:The total effective rate was 91.7% in the acupuncture-moxibustion group and 61.1% in the acupuncture group. The difference of the total effective rate between the two groups was statistically significant (P<0.05). After treatment, the scores of the clinical symptoms in the two groups were obviously decreased than those of the same group before treatment (P<0.01). The differences in the scores between the two groups were statistically significant (P<0.05). Conclusion:Acupuncture-moxibustion treatment to support yang and consolidate constitution is better than the routine acupuncture therapy in the therapeutic effect for CSA.