Objective:To explore the relationship between spindle and kinetochore-associated protein 3(SKA3), dual-specificity phosphatase 26(DUSP26), and prognosis in elderly patients with non-small cell lung cancer(NSCLC).Methods:A retrospective analysis was conducted on case samples from elderly patients with NSCLC at Xinxiang Central Hospital between January 2020 and May 2023.During surgery, specimens of cancerous and adjacent non-cancerous tissues were collected.The expressions of SKA3 and DUSP26 in these tissues were assessed using immunohistochemistry, and their correlations with clinicopathological characteristics were analyzed.The relationship between SKA3 and DUSP26 in cancer tissues was examined using the Spearman correlation coefficient.After one year of follow-up, the association between SKA3 and DUSP26 expressions in cancer tissues and patient prognosis was evaluated using Kaplan-Meier curves, and prognostic factors were analyzed using the Cox proportional hazards model.Results:In this cohort of 145 elderly patients aged 65 to 85 years(mean age: 72.61±3.87), including 91 males, we observed that the positive expression rates of SKA3 and DUSP26 in cancer tissues were 66.21%(96/145)and 71.03%(103/145), respectively.These rates were significantly higher than those found in para-carcinoma tissues, which were 16.55%(24/145)and 13.79%(20/145), with a P-value of <0.05.Spearman correlation analysis revealed a positive correlation between SKA3 and DUSP26 expression in cancer tissues( r=0.571, P<0.001).Moreover, the proportions of low differentiation, clinical staging at stages Ⅰ-Ⅱ, and lymph node metastasis were significantly higher in the SKA3-positive group compared to the SKA3-negative group( P<0.05), and similarly, these proportions were higher in the DUSP26-positive group than in the DUSP26-negative group( P<0.05).Kaplan-Meier survival analysis indicated that after one year of follow-up, the cumulative survival rates for patients with positive expressions of SKA3 and DUSP26 were 61.46%(59/96)and 58.25%(60/103), respectively, which were significantly lower than those with negative expressions[87.76%(43/49)and 92.86%(39/42), P<0.05].Cox regression analysis identified low differentiation( HR=1.817, 95% CI: 1.294-2.550), clinical staging at stage Ⅲ( HR=1.939, 95% CI: 1.315-2.858), lymph node metastasis( HR=1.898, 95% CI: 1.350-2.670), as well as positive expressions of SKA3( HR=2.071, 95% CI: 1.317-3.257)and DUSP26( HR=2.136, 95% CI: 1.402-3.256)as significant risk factors for prognosis( P<0.05). Conclusions:The expression rates of SKA3 and DUSP26 in cancer tissues are significantly elevated in elderly patients with NSCLC.Furthermore, these two biomarkers are correlated with the degree of differentiation, clinical staging, and lymph node metastasis, indicating their potential utility in evaluating the prognosis of elderly NSCLC patients.

Objective To compare the hematological parameters,biochemical profiles,histopathological characteristics,and cytokines associated with lipid metabolism in 8-weeks-old specific pathogen-free(SPF)grade and germ-free(GF)golden hamsters.Methods Twenty 8-weeks-old SPF grade and GF golden hamsters,with equal numbers of males and females,were utilized in this study.Serum cytokines associated with routine blood parameters,blood biochemistry,and lipid metabolism were quantified using automated hematology and biochemical analyzers,and enzyme-linked immunosorbent assay kits.The cecum,small intestine,lung,spleen,forestomach,and glandular stomach were examined by histopathology.Results We compared the result between SPF grade and GF golden hamsters at 8 weeks of age.Regarding hematological parameters,females showed significant differences(P＜0.05)in white blood cell count,hemoglobin concentration,mean corpuscular hemoglobin(MCH),and mean platelet volume between the two groups,and highly significant differences(P＜0.001)in MCH concentration(MCHC),platelet distribution width(PDW),and lymphocyte count(LYM).Males showed significant differences(P＜0.05)in mean corpuscular volume,MCH,LYM,neutrophil count,basophil count,and basophil percentage,and highly significant differences(P＜0.001)in MCHC,PDW,lymphocyte percentage,and neutrophil percentage.In terms of biochemical parameters,females showed significant differences(P＜0.05)in low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol,fasting insulin(FINS),and glycosylated serum protein(GSP),and highly significant differences(P＜0.001)in triglycerides(TG).Males showed significant differences(P＜0.05)in GSP levels and highly significant differences(P＜0.001)in total cholesterol,TG,LDL-C,FINS,and C-peptide levels.For serum cytokines related to lipid metabolism,females showed significant differences(P＜0.05)in interleukin(IL)-10,adiponectin(ADP),and IL-6 levels,and highly significant differences(P＜0.001)in high-sensitivity C-reactive protein(hs-CRP).Males showed significant differences(P＜0.05)in hs-CRP levels and highly significant differences(P＜0.001)in ADP and tumor necrosis factor-alpha levels.Hematoxylin/eosin staining showed that the cecal muscle layer was thinner and the number of crypts attached to the mucous membrane was reduced in GF compared with SPF grade golden hamsters.In addition,the ileocele was enlarged and the number of goblet cells was increased,the alveolar septum was widened,immune cells in the white pulp of the spleen were increased,and the blood content in the blood sinuses was increased.There was also thinning of the anterior gastric mucosa and the basophilic strength of the glandular gastric tube gland was weakened.Conclusions This study established the differences in routine blood parameters,blood biochemistry indicators,histopathological features,and cytokines associated with lipid metabolism between 8-weeks-old SPF grade and GF golden hamsters,to establish preliminary reference ranges.

In recent years,germ-free rats and mice have received extensive attention and application in the field of biomedical research,particularly in gut microbiota studies.The demand for germ-free rodents has been increasing,requiring many research institutions to establish quality germ-free animal populations.The process of breeding first-generation germ-free animals involves highly specialized and technically challenging work,such as using artificial rearing techniques to feed rodent pups.This review article provides an overview of key aspects of artificial rearing techniques,including nursing method,preparation of artificial milk,and sterilization method.It retrospectively summarizes the key technical points and,based on this foundation,offers prospects for further applications of artificial rearing techniques.

In recent years,germ-free rats and mice have received extensive attention and application in the field of biomedical research,particularly in gut microbiota studies.The demand for germ-free rodents has been increasing,requiring many research institutions to establish quality germ-free animal populations.The process of breeding first-generation germ-free animals involves highly specialized and technically challenging work,such as using artificial rearing techniques to feed rodent pups.This review article provides an overview of key aspects of artificial rearing techniques,including nursing method,preparation of artificial milk,and sterilization method.It retrospectively summarizes the key technical points and,based on this foundation,offers prospects for further applications of artificial rearing techniques.

Objective To compare the hematological parameters,biochemical profiles,histopathological characteristics,and cytokines associated with lipid metabolism in 8-weeks-old specific pathogen-free(SPF)grade and germ-free(GF)golden hamsters.Methods Twenty 8-weeks-old SPF grade and GF golden hamsters,with equal numbers of males and females,were utilized in this study.Serum cytokines associated with routine blood parameters,blood biochemistry,and lipid metabolism were quantified using automated hematology and biochemical analyzers,and enzyme-linked immunosorbent assay kits.The cecum,small intestine,lung,spleen,forestomach,and glandular stomach were examined by histopathology.Results We compared the result between SPF grade and GF golden hamsters at 8 weeks of age.Regarding hematological parameters,females showed significant differences(P＜0.05)in white blood cell count,hemoglobin concentration,mean corpuscular hemoglobin(MCH),and mean platelet volume between the two groups,and highly significant differences(P＜0.001)in MCH concentration(MCHC),platelet distribution width(PDW),and lymphocyte count(LYM).Males showed significant differences(P＜0.05)in mean corpuscular volume,MCH,LYM,neutrophil count,basophil count,and basophil percentage,and highly significant differences(P＜0.001)in MCHC,PDW,lymphocyte percentage,and neutrophil percentage.In terms of biochemical parameters,females showed significant differences(P＜0.05)in low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol,fasting insulin(FINS),and glycosylated serum protein(GSP),and highly significant differences(P＜0.001)in triglycerides(TG).Males showed significant differences(P＜0.05)in GSP levels and highly significant differences(P＜0.001)in total cholesterol,TG,LDL-C,FINS,and C-peptide levels.For serum cytokines related to lipid metabolism,females showed significant differences(P＜0.05)in interleukin(IL)-10,adiponectin(ADP),and IL-6 levels,and highly significant differences(P＜0.001)in high-sensitivity C-reactive protein(hs-CRP).Males showed significant differences(P＜0.05)in hs-CRP levels and highly significant differences(P＜0.001)in ADP and tumor necrosis factor-alpha levels.Hematoxylin/eosin staining showed that the cecal muscle layer was thinner and the number of crypts attached to the mucous membrane was reduced in GF compared with SPF grade golden hamsters.In addition,the ileocele was enlarged and the number of goblet cells was increased,the alveolar septum was widened,immune cells in the white pulp of the spleen were increased,and the blood content in the blood sinuses was increased.There was also thinning of the anterior gastric mucosa and the basophilic strength of the glandular gastric tube gland was weakened.Conclusions This study established the differences in routine blood parameters,blood biochemistry indicators,histopathological features,and cytokines associated with lipid metabolism between 8-weeks-old SPF grade and GF golden hamsters,to establish preliminary reference ranges.

Objective:To explore the relationship between spindle and kinetochore-associated protein 3(SKA3), dual-specificity phosphatase 26(DUSP26), and prognosis in elderly patients with non-small cell lung cancer(NSCLC).Methods:A retrospective analysis was conducted on case samples from elderly patients with NSCLC at Xinxiang Central Hospital between January 2020 and May 2023.During surgery, specimens of cancerous and adjacent non-cancerous tissues were collected.The expressions of SKA3 and DUSP26 in these tissues were assessed using immunohistochemistry, and their correlations with clinicopathological characteristics were analyzed.The relationship between SKA3 and DUSP26 in cancer tissues was examined using the Spearman correlation coefficient.After one year of follow-up, the association between SKA3 and DUSP26 expressions in cancer tissues and patient prognosis was evaluated using Kaplan-Meier curves, and prognostic factors were analyzed using the Cox proportional hazards model.Results:In this cohort of 145 elderly patients aged 65 to 85 years(mean age: 72.61±3.87), including 91 males, we observed that the positive expression rates of SKA3 and DUSP26 in cancer tissues were 66.21%(96/145)and 71.03%(103/145), respectively.These rates were significantly higher than those found in para-carcinoma tissues, which were 16.55%(24/145)and 13.79%(20/145), with a P-value of <0.05.Spearman correlation analysis revealed a positive correlation between SKA3 and DUSP26 expression in cancer tissues( r=0.571, P<0.001).Moreover, the proportions of low differentiation, clinical staging at stages Ⅰ-Ⅱ, and lymph node metastasis were significantly higher in the SKA3-positive group compared to the SKA3-negative group( P<0.05), and similarly, these proportions were higher in the DUSP26-positive group than in the DUSP26-negative group( P<0.05).Kaplan-Meier survival analysis indicated that after one year of follow-up, the cumulative survival rates for patients with positive expressions of SKA3 and DUSP26 were 61.46%(59/96)and 58.25%(60/103), respectively, which were significantly lower than those with negative expressions[87.76%(43/49)and 92.86%(39/42), P<0.05].Cox regression analysis identified low differentiation( HR=1.817, 95% CI: 1.294-2.550), clinical staging at stage Ⅲ( HR=1.939, 95% CI: 1.315-2.858), lymph node metastasis( HR=1.898, 95% CI: 1.350-2.670), as well as positive expressions of SKA3( HR=2.071, 95% CI: 1.317-3.257)and DUSP26( HR=2.136, 95% CI: 1.402-3.256)as significant risk factors for prognosis( P<0.05). Conclusions:The expression rates of SKA3 and DUSP26 in cancer tissues are significantly elevated in elderly patients with NSCLC.Furthermore, these two biomarkers are correlated with the degree of differentiation, clinical staging, and lymph node metastasis, indicating their potential utility in evaluating the prognosis of elderly NSCLC patients.

Chaihu Shugansan composed of Bupleuri Radix， Paeoniae Radix Alba， Chuanxiong Rhizoma， Aurantii Fructus， Citri Reticulatae Pericarpium， Cyperi Rhizoma， and Glycyrrhizae Radix et Rhizoma has the effects of soothing liver， relieving depression， regulating Qi movement， and relieving pain. It is a classic formula for treating gastric distension recommended by doctors of later ages. This article systematically reviews the clinical application and basic experimental progress of Chaihu Shugansan in the treatment of functional dyspepsia. In modern clinical practice， Chaihu Shugansan and its modified formulas are used to treat functional dyspepsia， and they can be applied in combination with other formulas （Si Junzitang， Jinlingzisan， Zhizhuwan， etc.）， western medicine （domperidone tablets， deanxit， Saccharomyces boulardii， etc.）， traditional Chinese medicine （TCM） acupuncture and other therapies. The results of clinical studies have shown that Chaihu Shugansan and its modified formulas can significantly reduce the Hamilton depression scale （HAMD） score， Hamilton anxiety scale （HAMA） score， and TCM syndrome score， ameliorate the symptoms， improve the quality of life， and decrease the recurrence rate. The experimental pharmacological studies have demonstrated that Chaihu Shugansan can inhibit the autophagy of Cajal interstitial cells， regulate the endoplasmic reticulum stress signaling pathway， and modulate the brain-gut peptide level to improve the gastrointestinal motility. Chaihu Shugansan can inhibit the expression of 5-hydroxytryptamine in the colon tissue and reduce the abdominal withdrawal reflex （AWR） score to improve visceral hypersensitivity. Furthermore， Chaihu Shugansan can lower the levels of pro-inflammatory cytokines such as interleukin-6 and tumor necrosis factor-α to repair duodenal mucosal inflammation. In addition， it can regulate intestinal flora to maintain intestinal flora balance. The main active ingredients such as saikosaponin， paeoniflorin， hesperidin， and naringin in Chaihu Shugansan can exert anti-inflammatory， antioxidant， and antimicrobial effects.

Objective A sterile golden hamster model was established by cesarean section purification.Methods SPF-grade donor female golden hamsters were selected,and males and females were mated 1∶1 and separated after mating.The cage time of the surrogate mothers was 1 week earlier than that of the donor mothers.Parturient golden hamsters underwent hysterectomy on a sterilized workbench,and the uteruses were transferred into isolation kits and stripped.To obtain sterile milk for milk replacement,sterile ICR mice and sterile SD rats were used.After successful separation,the hamsters were transferred to isolation kits to prepare for feeding.The sterility status of the feeding isolation kits was tested monthly.Results Three caesarean sections were performed,but the first and second lactations failed.The third milk replacement was successful,and 18 young hamsters were obtained with survival rates of 88%and 66%after weaning.All hamsters were quality tested by GB/T 14926.41-2001.Conclusions Using a cesarean section purification technique and sterile ICR mice and SD rats for microbial-free milk replacement,a sterile golden hamster model was obtained.

Objective To evaluate the effects of fecal status and transplantation method on the intestinal flora structure after fecal microbiota transplantation in germ-free mice.Methods Thirty-six C57BL/6 mice were divided randomly into three groups:fresh fecal gavage transplantation(group A),frozen fecal gavage transplantation(group B),and frozen fecal rectal transplantation groups(group C).Feces were collected at 2,4 and 6 weeks after transplantation.All mice were sacrificed at 6 weeks to obtain the contents of the small and large intestines.The structure and function of the gut microbiota and dynamic trends in microbial changes were analyzed by 16 S rRNA sequencing.Results The diversity of the small intestine microbes in both the group A and group C were similar to those in the group B,according to α-diversity analysis(P＞0.05),but the diversity of large intestine microbes was significantly increased(P＜0.001).According toβ-diversity analysis,small intestine samples from the group A and group B clustered in the same area,indicating that the microbial community compositions were similar(P＞0.05),but samples from the large intestine were distributed in different areas,showing significant differences(P＜0.001).Small and large intestine samples from the group B and group C were distributed in different areas,with significant differences(P＜0.001).Linear discriminant analysis effect size showed that Bacteroidota were relatively dominant in the group A,while Verrucomicrobiota and Proteobacteria were relatively dominant in the group B and Firmicutes were relatively dominant in the group C.Functional prediction using PICRUSt2 software showed that neither fecal status nor the method of transplantation affected the functions of the microbial community.Conclusions Both fresh fecal gavage and frozen fecal rectal transplantation can enhance the microbial diversity of the large intestine,compared with frozen fecal gavage transplantation.Fecal status does not affect the gut function and colonization trends of the microbiota,whereas the method of transplantation affects the colonization trends but not the functions of the microbiota.

Objective To evaluate the effects of fecal status and transplantation method on the intestinal flora structure after fecal microbiota transplantation in germ-free mice.Methods Thirty-six C57BL/6 mice were divided randomly into three groups:fresh fecal gavage transplantation(group A),frozen fecal gavage transplantation(group B),and frozen fecal rectal transplantation groups(group C).Feces were collected at 2,4 and 6 weeks after transplantation.All mice were sacrificed at 6 weeks to obtain the contents of the small and large intestines.The structure and function of the gut microbiota and dynamic trends in microbial changes were analyzed by 16 S rRNA sequencing.Results The diversity of the small intestine microbes in both the group A and group C were similar to those in the group B,according to α-diversity analysis(P＞0.05),but the diversity of large intestine microbes was significantly increased(P＜0.001).According toβ-diversity analysis,small intestine samples from the group A and group B clustered in the same area,indicating that the microbial community compositions were similar(P＞0.05),but samples from the large intestine were distributed in different areas,showing significant differences(P＜0.001).Small and large intestine samples from the group B and group C were distributed in different areas,with significant differences(P＜0.001).Linear discriminant analysis effect size showed that Bacteroidota were relatively dominant in the group A,while Verrucomicrobiota and Proteobacteria were relatively dominant in the group B and Firmicutes were relatively dominant in the group C.Functional prediction using PICRUSt2 software showed that neither fecal status nor the method of transplantation affected the functions of the microbial community.Conclusions Both fresh fecal gavage and frozen fecal rectal transplantation can enhance the microbial diversity of the large intestine,compared with frozen fecal gavage transplantation.Fecal status does not affect the gut function and colonization trends of the microbiota,whereas the method of transplantation affects the colonization trends but not the functions of the microbiota.