Objective To clone and express the heat shock cognate protein 20 (SjHsc20) of Schistosoma japonicum, and to preliminarily investigate its biological characteristics. Methods The target fragment of the SjHsc20 gene was amplified using PCR assay and cloned into the pET-28a(+) expression plasmid to generate the recombinant expression vector pET-28a(+)-SjH-sc20, which was then transformed into Escherichia coli BL21 (DE3) competent cells. The recombinant SjHsc20 (rSjHsc20) protein was induced with isopropyl β-D-thiogalactopyranoside (IPTG) and purified, and the expression of the rSjHsc20 protein was checked with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The immunogenicity of the rSjHsc20 protein was detected using Western blotting, and the transcriptional levels of SjHsc20 were quantified in S. japonicum worms at different developmental stages and in male and female adult worms using real-time quantitative PCR (RT-qPCR) assay. Thirty female BALB/c mice at ages 6 to 8 weeks were divided into three groups, including the rSjHsc20 immunization group, the PBS control group, and the ISA 206 adjuvant group, of 10 mice in each group. Mice in the rSjHsc20 immunization group were subcutaneously immunized with 20 μg rSjHsc20 on days 1, 15 and 31, and animals in the PBS control group were subcutaneously injected with the same volume of PBS on days 1, 15 and 31, while mice in the ISA 206 adjuvant group were subcutaneously immunized with the same volume of ISA 206 adjuvant on days 1, 15 and 31, respectively. All mice in each group were infected with (40 ± 2) S. japonicum cercariae via the abdomen 14 day following the last immunization. Levels of serum specific IgG and its subtypes IgG1 and IgG2 antibodies against rSjHsc20, and the serum titers of anti-rSjHsc20 antibody were detected in mice using indirect enzyme-linked immunosorbent assay (ELISA). All mice were sacrifice 42 days post-infection, and S. japonicum worms were collected from the hepatic portal vein and counted. The eggs per gram (EPG), worm burden reductions and egg burden reductions were estimated to evaluate the protective efficacy of the rSjHsc20 protein. Results The SjHsc20 gene had an open reading frame (ORF) with 756 bp in length and encoded 252 amino acids, and the rSjHsc20 protein had a relative molecular mass of approximately 29 kDa. The rSjHsc20 protein was recognized by the serum of mice infected with S. japonicum and the serum of mice immunized with the rSjHsc20 protein, indicating that rSjHsc20 had a good immunogenicity. There was a significant difference in the transcriptional levels of the SjHsc20 gene among the 7-day (1.001 4 ± 0.065 7), 12-day (2.268 3 ± 0.129 2), 21-day (1.378 5 ± 0.160 4), 28-day (1.196 4 ± 0.244 0), 35-day (1.646 3 ± 0.226 1), 42-day worms of S. japonicum (1.758 0 ± 0.611 1) (F = 38.45, P < 0.000 1), and the transcriptional level of the SjHsc20 gene was higher in the 12-day worms than in worms at other developmental stages (all P values < 0.000 1). The serum levels of anti-rSjHsc20 IgG antibody were 0.106 6 ± 0.010 7, 0.108 3 ± 0.010 4, and 0.553 2 ± 0.069 1 in the PBS control group, ISA 206 adjuvant group, and rSjHsc20 immunization group following the last immunization, respectively, and the serum levels of IgG1 antibody were 0.137 3 ± 0.054 0, 0.181 1 ± 0.096 8, and 1.765 8 ± 0.221 1, while the levels of IgG2a antibody were 0.280 3 ± 0.197 6, 0.274 0 ± 0.146 3, and 1.560 4 ± 0.106 0, respectively. There were significant differences in the serum levels of anti-rSjHsc20 IgG (F = 397.70, P < 0.000 1), IgG1 (F = 401.00, P < 0.000 1) and IgG2a antibodies (F = 229.70, P < 0.000 1) among the three groups, and the serum levels of anti-rSjHsc20 IgG, IgG1 and IgG2a antibodies were higher in the rSjHsc20 immunization group than in the PBS control group and the ISA 206 adjuvant group (all P values < 0.000 1). There was a significant difference in the IgG1/IgG2a ratio among the rSjHsc20 immunization group (1.177 2 ± 0.143 6), the PBS control group (0.428 4 ± 0.199 8) and the ISA 206 adjuvant group (0.559 9 ± 0.181 1) (F = 43.97, P < 0.000 1), and the IgG1/IgG2a ratio was > 1 in the rSjHsc20 immunization group, which was higher than in the PBS control group and the ISA 206 adjuvant group (both P values < 0.000 1). The titers of serum anti-rSjHsc20 antibody were all above 1∶16 384 in the rSjHsc20 immunization group following immunizations on days 1, 15 and 31, indicating that the rSjHsc20 protein had a strong immunogenicity. The mean worm burdens were (16.60±5.75), (15.80±5.58) worms per mouse and (14.40±5.75) worms per mouse in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group 42 days post-infection with S. japonicum cercariae (F = 0.50, P > 0.05), and the EPG were 68 370 ± 22 690, 67 972 ± 19 502, and 41 075 ± 13 251 in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group (F = 4.55, P < 0.05), with lower EPG in the PBS control group and the ISA 206 adjuvant group than in the rSjHsc20 immunization group (both P values < 0.05). Immunization with the rSjHsc20 protein resulted in a worm burden reduction of 13.25% and an egg burden reduction of 39.92% relative to the PBS control group. Conclusions SjHsc20 is successfully cloned and expressed, and the rSjHsc20 protein induces partial immunoprotective effects in mice, which provides a basis for deciphering the biological functions of SjHsc20 and assessing the potential of SjH-sc20 as a vaccine candidate.

BACKGROUND:Electroacupuncture can treat oligoasthenospermia by stimulating hormone synthesis and release in the hypothalamus,pituitary gland and testis,but its mechanism is complex,and the efficacy of electroacupuncture is closely related to the idea of acupoint compatibility.OBJECTIVE:To explore whether electroacupuncture therapy can improve the function status of hypothalamic-pituitary-testicular axis by adjusting the level of serum related sex hormones in rats,so as to treat adenine-induced oligoasthenospermia in rats.METHODS:Fifty adult male Sprague-Dawley rats were randomly divided into blank,model,electroacupuncture,L-carnitine,and non-meridian and non-acupuncture point groups,with 10 rats in each group.Except for the blank group,the rats in the other four groups were given 200 mg/(kg·d)adenine via intragastric administration for 28 days to establish oligoasthenospermia models.After modeling,the rats in the electroacupuncture group were given electroacupuncture treatment at Zhongji,Guanyuan,Zusanli and Sanyinjiao acupoints,the non-acupoint group was selected for electroacupuncture treatment at three non-acupoint points,and the rats in the L-carnitine group were given oral infusion of L-carnitine,once daily,for 28 continuous days.After treatment,the kidney and testicular organ coefficients were calculated.Sperm count,survival rate and motility rate were determined.Pathological morphological changes of kidney and testicular tissues were observed.Serum sex hormones testosterone,gonadotropin releasing hormone,follicle stimulating hormone,luteinizing hormone,estradiol,statin B and prolactin levels were detected in rats of each group.RESULTS AND CONCLUSION:(1)The renal organ coefficient in the model group was higher than that in the blank,electroacupuncture and L-carnitine groups(P＜0.05),the renal organ coefficient in the non-acupoint group was higher than that in the electroacupuncture group(P＜0.05),and there was no significant difference in the testicular organ coefficient among all groups(P＞0.05).(2)Compared with the blank group,sperm count,survival rate and motility rate were decreased in the model group(P＜0.05);compared with the model group,sperm count,survival rate and motility rate were increased in the electroacupuncture group and L-carnitine group(P＜0.05);compared with the non-acupoint group,sperm count,survival rate and motility rate were higher in the electroacupuncture group(P＜0.05).(3)Hematoxylin-eosin staining results indicated that the kidney and testicular tissues of rats in the model group were severely damaged,while damage to kidney and testicular tissues was less in the electroacupuncture group and L-carnitine group compared with the model group,but severer in the non-acupoint group than the electroacupuncture group.(4)Compared with the blank group,the levels of testosterone,gonadotropin releasing hormone,and inhibin B in the serum of rats in the model group were decreased(P＜0.05),and the levels of follicle-stimulating hormone,luteinizing hormone,estradiol,and prolactin were increased(P＜0.05).Compared with the model group,the levels of testosterone,gonadotropin-releasing hormone,and inhibin B in the serum of rats in the electroacupuncture group and L-carnitine group were increased(P＜0.05),and the levels of follicle-stimulating hormone,luteinizing hormone,estradiol,and prolactin were decreased(P＜0.05).Compared with the non-acupoint group,the levels of testosterone,gonadotropin-releasing hormone,and inhibin Bin the serum of the rats in electroacupuncture group were increased(P＜0.05),and the levels of follicle-stimulating hormone,luteinizing hormone,estradiol,and prolactin were decreased(P＜0.05).To conclude,electroacupuncture at Zhongji,Guanyuan,Zusanli,and Sanyinjiao acupoints can effectively treat oligoasthenospermia by regulating the functional state of the hypothalamic-pituitary-testicular axis in rats.

BACKGROUND:Electroacupuncture can treat oligoasthenospermia by stimulating hormone synthesis and release in the hypothalamus,pituitary gland and testis,but its mechanism is complex,and the efficacy of electroacupuncture is closely related to the idea of acupoint compatibility.OBJECTIVE:To explore whether electroacupuncture therapy can improve the function status of hypothalamic-pituitary-testicular axis by adjusting the level of serum related sex hormones in rats,so as to treat adenine-induced oligoasthenospermia in rats.METHODS:Fifty adult male Sprague-Dawley rats were randomly divided into blank,model,electroacupuncture,L-carnitine,and non-meridian and non-acupuncture point groups,with 10 rats in each group.Except for the blank group,the rats in the other four groups were given 200 mg/(kg·d)adenine via intragastric administration for 28 days to establish oligoasthenospermia models.After modeling,the rats in the electroacupuncture group were given electroacupuncture treatment at Zhongji,Guanyuan,Zusanli and Sanyinjiao acupoints,the non-acupoint group was selected for electroacupuncture treatment at three non-acupoint points,and the rats in the L-carnitine group were given oral infusion of L-carnitine,once daily,for 28 continuous days.After treatment,the kidney and testicular organ coefficients were calculated.Sperm count,survival rate and motility rate were determined.Pathological morphological changes of kidney and testicular tissues were observed.Serum sex hormones testosterone,gonadotropin releasing hormone,follicle stimulating hormone,luteinizing hormone,estradiol,statin B and prolactin levels were detected in rats of each group.RESULTS AND CONCLUSION:(1)The renal organ coefficient in the model group was higher than that in the blank,electroacupuncture and L-carnitine groups(P＜0.05),the renal organ coefficient in the non-acupoint group was higher than that in the electroacupuncture group(P＜0.05),and there was no significant difference in the testicular organ coefficient among all groups(P＞0.05).(2)Compared with the blank group,sperm count,survival rate and motility rate were decreased in the model group(P＜0.05);compared with the model group,sperm count,survival rate and motility rate were increased in the electroacupuncture group and L-carnitine group(P＜0.05);compared with the non-acupoint group,sperm count,survival rate and motility rate were higher in the electroacupuncture group(P＜0.05).(3)Hematoxylin-eosin staining results indicated that the kidney and testicular tissues of rats in the model group were severely damaged,while damage to kidney and testicular tissues was less in the electroacupuncture group and L-carnitine group compared with the model group,but severer in the non-acupoint group than the electroacupuncture group.(4)Compared with the blank group,the levels of testosterone,gonadotropin releasing hormone,and inhibin B in the serum of rats in the model group were decreased(P＜0.05),and the levels of follicle-stimulating hormone,luteinizing hormone,estradiol,and prolactin were increased(P＜0.05).Compared with the model group,the levels of testosterone,gonadotropin-releasing hormone,and inhibin B in the serum of rats in the electroacupuncture group and L-carnitine group were increased(P＜0.05),and the levels of follicle-stimulating hormone,luteinizing hormone,estradiol,and prolactin were decreased(P＜0.05).Compared with the non-acupoint group,the levels of testosterone,gonadotropin-releasing hormone,and inhibin Bin the serum of the rats in electroacupuncture group were increased(P＜0.05),and the levels of follicle-stimulating hormone,luteinizing hormone,estradiol,and prolactin were decreased(P＜0.05).To conclude,electroacupuncture at Zhongji,Guanyuan,Zusanli,and Sanyinjiao acupoints can effectively treat oligoasthenospermia by regulating the functional state of the hypothalamic-pituitary-testicular axis in rats.

【Objective】 To investigate the association between genetic variations in the glucagon-like peptide-1 receptor (GLP-1R) gene and BP responses to sodium and potassium intake. 【Methods】 A total of 514 subjects from 124 families were recruited in Meixian County, Shaanxi Province, in 2004, resulting in the establishment of a "salt-sensitive hypertension study cohort" . The subjects followed a dietary regimen which involved a normal diet for 3 days, a low-salt diet for 7 days, a high-salt diet for 7 days, and a high-salt potassium-supplemented diet for 7 days. BP measurement was conducted at different intervention periods, and peripheral blood samples were collected. Additionally, eight single nucleotide polymorphisms (SNPs) of the GLP-1R gene were genotyped using the MassARRAY detection platform. 【Results】 The GLP-1R gene SNP rs9462472 exhibited a significant association with systolic BP, diastolic BP, and mean arterial pressure response to high-salt intervention. Similarly, SNP rs2268637 showed a significant association with systolic BP response to high-salt intervention. Furthermore, SNP rs2268637 was significantly associated with systolic BP and mean arterial pressure responses to high-salt plus potassium supplementation intervention. 【Conclusion】 Our findings indicate a significant association of genetic variations in the GLP-1R gene with BP responses to sodium and potassium intake. This suggests that the GLP-1R gene plays a role in the regulation of BP salt sensitivity and potassium sensitivity.

[摘 要] 目的：研究芳香烃受体（AHR）在乳腺癌中的表达及其对乳腺癌细胞增殖、凋亡和药物敏感性的调控机制。方法：通过GEPIA数据库数据分析乳腺癌组织及癌旁组织中AHR的表达水平，探讨其与患者生存期的关联。利用基因敲低和过表达技术构建AHR表达变化的乳腺癌细胞，采用CCK-8实验、细胞计数和流式细胞分析等方法评估AHR对细胞增殖、凋亡和药物敏感性的影响，通过免疫印迹法验证相关分子机制。此外，利用AHR激动剂6-甲酰基吲哚并[3,2-B]咔唑（FICZ）研究外源性激活AHR对乳腺癌细胞多柔比星（DOX）敏感性的影响。结果：GEPIA数据库数据分析结果显示，乳腺癌组织中AHR呈明显低表达（P < 0.05）；对155例乳腺癌患者的生存期进行统计分析也显示AHR低表达与不良预后呈正相关（P < 0.05）。敲低AHR促进细胞增殖（P < 0.05），过表达则能抑制其增殖（P < 0.05）并促进其凋亡（P < 0.05）。外源激活AHR能增强乳腺癌细胞对DOX的敏感性（P < 0.05）。AHR可与MYC基因启动子结合，抑制MYC表达（P < 0.05），从而影响乳腺癌的进展。结论：AHR在乳腺癌中通过调控MYC表达影响细胞增殖和凋亡，外源激活AHR可能成为提高乳腺癌细胞对DOX敏感性的治疗策略。

Objective Experimental animal facilities account for a significant proportion of the energy consumption by scientific research institutions;however,the energy consumption characteristics of these facilities differ from those of ordinary buildings,and thus require specialized monitoring and management.Methods A set of energy consumption monitoring systems was designed for experimental animal facilities and deployed in the specific pathogen-free-level experimental animal facility of Huazhong University of Science and Technology.Results The system achieved real-time collection and recording of the facility's electricity consumption data,and proposed energy-saving measures for three application scenarios.Conclusions This energy consumption monitoring system designed for experimental animal facilities is reliable,efficient,and user-friendly,and has the potential to guide and promote energy management programs at experimental animal facilities.

Objective:To explore the relationship between glycosylated hemoglobin A 1c/high-density lipoprotein cholesterol ratio (HbA 1c/HDL-C) and urinary albumin-creatinine ratio (UACR) in Chinese adults. Methods:In this cross-sectional study, the clinical data of 43 820 community residents (age>40 years) from the Risk Evaluation of Cancers in Chinese Diabetic Individuals (REACTION study; March-December 2012) across eight centers (Liaoning, Guangdong, Shanghai, Gansu, Guangxi, Henan, Hubei, and Sichuan) in China were collected and analyzed. Participants were divided into three groups based on UACR levels:<10 mg/g, 10-30 mg/g, and >30 mg/g. The HbA 1c/HDL-C ratio was divided into four groups according to quartile division of the subjects: 1st quartile (Q1<3.79), 2nd quartile (3.79≤Q2<4.59), 3rd quartile (4.59≤Q3≤5.66), and 4th quartile (Q4>5.66). Multivariate ordinal logistic regression model was used to analyze the relationship between HbA 1c/HDL-C and UACR. Receiver operating characteristic (ROC) analysis was used to explore the predictive value of HbA 1c/HDL-C to UACR. Results:The 43 820 subjects included 13 452 (30.70%) male and 30 378 (69.30%) female patients, with an average age of (58.00±0.05) years. According to results of one-way analysis of variance analysis, the HbA 1c/HDL-C ratio was significantly associated with the risk of increased UACR ( F=495.73, P<0.001). After adjusting for clinically relevant confounding variables in logistic regression model, compared with participants with the lowest HbA 1c/HDL-C ratio (Q1), women with the highest HbA 1c/HDL-C ratio (Q4) had a 1.483-fold (95% CI 1.376-1.598, P<0.001) and men had a 1.161-fold (95% CI 1.019-1.323, P<0.001) increased risk of UACR. The ROC curve analysis showed that the area under the curve of HbA 1c/HDL-C for predicting increased UACR was 0.623 (95% CI 0.597-0.606), with a sensitivity of 60.18% and a specificity of 54.91%. The HbA 1c/HDL-C ratio showed the highest predictive value of all glycemic and lipidemic parameters. In individuals with well-controlled blood glucose (HbA 1c<6.5%) or lipid levels (HDL-C≥1.0 mmol/L), the HbA 1c/HDL-C ratio was still independently associated with the risk of increased UACR after adjusting for confounding variables [ OR(95% CI) of quartile 4: 1.563 (1.210-2.019, P=0.001) in participants with HbA 1c<6.5% and 1.822 (1.687-1.968, P<0.001) in participants with HDL-C≥1.0 mmol/L]. Conclusion:As a novel compound indicator for evaluating glucose homeostasis and dyslipidemia, the HbA 1c/HDL-C ratio was independently associated with increased UACR in the general population aged>40 years in China, which was superior to both glycemic and lipid parameters alone.

Objective:To explore the application value of qualitative characteristics and quantitative parameters of contrast-enhanced ultrasound (CEUS) in the differential diagnosis of pancreatic ductal adenocarcinoma (PDAC) and non-PDAC presenting as pancreatic solid focal lesions.Methods:A retrospective analysis was conducted on 64 cases of PDAC(the PDAC group) and 52 cases of non-PDAC(the non-PDAC group) who underwent CEUS examination at Tianjin Medical University Cancer Institute and Hospital from July 2022 to June 2023. Clinical characteristics, two-dimensional ultrasound features, CEUS qualitative characteristic, and quantitative parameters were compared between the two groups. ROC curves were plotted, and the Delong test was used to evaluate the diagnostic performance of qualitative and quantitative analyses in distinguishing PDAC from non-PDAC. Binary logistic regression analysis was employed to assess the independent predictors of PDAC.Results:①There were significant differences in serum CA19-9, lesion size, boundary, the main pancreatic duct (MPD) diameter, degree of enhancement and enhancement pattern between the PDAC group and the non-PDAC group (all P<0.05). ②The relative peak intensity (rPE), and relative wash-in and wash-out area under the curve (rWiWoAUC) were lower in the PDAC group than the non-PDAC group, with statistically significant differences(all P<0.001). ③The areas under the curve (AUC) for diagnosing PDAC using enhancement pattern, venous phase(VP) enhancement degree, rPE, and rWiWoAUC were 0.698, 0.707, 0.863, and 0.867, respectively. The AUCs of quantitative parameters were superior to those of qualitative characteristics, with statistically significant differences ( P<0.05). Using CEUS mode B, low VP enhancement, rPE<72.44, and rWiWoAUC<86.59 as cut-off values, the accuracies for diagnosing PDAC were 0.698, 0.741, 0.828, and 0.802, respectively. ④Serum CA19-9, lesion size, MPD diameter, rPE, and rWiWoAUC were independent predictors of PDAC (all P<0.05). Conclusions:CEUS qualitative and quantitative analyses are helpful in the differential diagnosis of PDAC and non-PDAC, with rPE and rWiWoAUC being useful indicators for diagnosing PDAC.

【Objective】 Dyslipidemia has shown to be associated with cardiovascular, metabolic and renal diseases. This study aimed to investigate the association between residual cholesterol and the risk of subclinical renal damage (SRD). 【Methods】 A total of 2 342 participants were recruited from the previously established Hanzhong Adolescent Hypertension Study cohort. According to estimated glomerular filtration rate(eGFR) and urinary albumin-to-creatine ratio(uACR), the subjects were divided into SRD group and non-SRD group. The associations of residual cholesterol with eGFR, uACR, and the risk of SRD were analyzed by multiple linear and Logistic regression analyses. 【Results】 Residual cholesterol was positively correlated with uACR(r=0.081, P<0.001) but negatively correlated with eGFR (r=-0.091, P<0.001). Multiple linear regression analysis revealed that residual cholesterol was an influencing factor of uACR (β=0.075, P<0.001) and eGFR (β=-0.027, P<0.001) after adjustment for gender, age, smoke, alcohol, exercise, BMI, hypertension, diabetes and serum uric acid. In addition, Logistic regression analysis revealed that residual cholesterol was significantly associated with the risk of SRD independently of potential confounders [OR(95% CI)=1.387 (1.113-1.728), P<0.001]. Further subgroup analysis showed that residual cholesterol was significantly associated with the risk of SRD in women but not in men. 【Conclusion】 Residual cholesterol is a contributing factor in the risk of subclinical renal damage with gender-specific association.

【Objective】 4-like protein with down-regulated expression and development in neural precursor cells (NEDD4L) plays an important role in blood pressure (BP) regulation and sodium homeostasis by regulating epithelial sodium channel protein. In this study, we aimed to explore the relationship of NEDD4L gene polymorphisms with BP responses to sodium and potassium intake. 【Methods】 In 2004, 514 subjects from 124 families in Meixian County, Shaanxi Province, were recruited to establish a salt-sensitive hypertension study cohort. All the subjects received a 3-day baseline survey, a 7-day low-salt diet, a 7-day high-salt diet, and finally a 7-day high-salt and potassium supplementation. Their BP was measured and peripheral blood samples were collected at different intervention periods. The 14 gene polymorphisms of NEDD4L gene were genotyped and analyzed by MassARRAY platform. 【Results】 BP decreased on a low-salt diet, and significantly increased on a high-salt diet, and decreased again after potassium supplementation. NEDD4L SNPs rs74408486 were significantly associated with systolic BP, diastolic BP and mean arterial pressure responses to the low-salt diet. SNPs rs292449 and rs2288775 were significantly associated with pulse pressure response to the high-salt diet. In addition, SNPs rs563283 and rs292449 were significantly associated with diastolic BP, mean arterial pressure, and pulse pressure responses to high-salt and potassium supplementation diet. 【Conclusion】 NEDD4L gene polymorphisms were significantly associated with BP responses to sodium and potassium intake, suggesting that NEDD4L gene may be involved in the development of salt sensitivity and potassium sensitivity.