Objective:To investigate the association between the local renin-angiotensin system (RAS) in human follicular fluid (hFF) and age, ovarian function, in vitro fertilization (IVF) laboratory outcome. Methods:A non-intervention observational study was designed. hFF and medical case history without personal identity of patients who received IVF simply because of male factor infertility in Reproductive Medicine Center, the First Affiliated Hospital of Soochow University during January 2021 and February 2022 were collected. The renin, angiotensin converting enzyme (ACE), ACE2, angiotensin (Ang)Ⅱ and Ang 1-7 levels were detected by enzyme-linked immunosorbent assay kit. The correlation between age and RAS in hFF was analyzed by simple linear regression, and multivariate linear regression was used to further analyze the correlation between the RAS and IVF laboratory outcome.Results:1) A total of 139 samples of analysable hFF were obtained. 2) There was a linear negative correlation between age and renin (Pearson's r=-0.313 3, P<0.001), angiotensin converting enzyme (ACE; Pearson's r=-0.183 6, P=0.031), angiotensinⅡ (AngⅡ; Pearson's r=-0.218 6, P=0.010), ACE/ACE2 (Pearson's r=-0.319 2, P<0.001), AngⅡ/Ang1-7 (Pearson's r=-0.224 3, P=0.008), while the linear relationship with ACE2 and Ang1-7 was not significant (all P>0.05). 3) Basal follicle-stimulating hormone was positively correlated with age ( β=0.636, P<0.001), ACE2 ( β=0.267, P=0.026) and AngⅡ ( β=0.268, P=0.001), while negatively correlated with ACE ( β=-0.320, P<0.001) and Ang1-7 ( β=-0.217, P=0.014). Basal luteinizing hormone was positively correlated with AngⅡ ( β=0.330, P=0.003), while negative correlated with Ang1-7 ( β=-0.395, P=0.002). Antral follicle count was positively correlated with Ang1-7 ( β=0.153, P=0.049), while negatively correlated with age ( β=-0.869, P<0.001) and ACE2 ( β=-0.082, P=0.004). Basal anti-Müllerian hormone was only negatively correlated with age ( β=-0.349, P<0.001). There was no correlation between RAS and basal estradiol, progesterone, prolactin, testosterone (all P>0.05). 4) Oocyte retrieval count was positively correlated with renin ( β=0.146, P=0.014), AngⅡ ( β=0.113, P=0.034) and Ang1-7 ( β=0.185, P=0.002), while negatively correlated with age ( β=-0.717, P<0.001); MⅡ oocyte maturation rate was positively correlated with AngⅡ ( β=0.207, P=0.019) and Ang1-7 ( β=0.217, P=0.026), while negatively correlated with age ( β=-0.518, P<0.001). There was no correlation between RAS and the rates of two pronuclei embryos, transplantable embryos, high-quality embryos (all P>0.05). Conclusion:The local RAS in ovarian follicles is affected by age and correlated with ovarian function and IVF laboratory outcome.

Objective:To investigate the association between the local renin-angiotensin system (RAS) in human follicular fluid (hFF) and age, ovarian function, in vitro fertilization (IVF) laboratory outcome. Methods:A non-intervention observational study was designed. hFF and medical case history without personal identity of patients who received IVF simply because of male factor infertility in Reproductive Medicine Center, the First Affiliated Hospital of Soochow University during January 2021 and February 2022 were collected. The renin, angiotensin converting enzyme (ACE), ACE2, angiotensin (Ang)Ⅱ and Ang 1-7 levels were detected by enzyme-linked immunosorbent assay kit. The correlation between age and RAS in hFF was analyzed by simple linear regression, and multivariate linear regression was used to further analyze the correlation between the RAS and IVF laboratory outcome.Results:1) A total of 139 samples of analysable hFF were obtained. 2) There was a linear negative correlation between age and renin (Pearson's r=-0.313 3, P<0.001), angiotensin converting enzyme (ACE; Pearson's r=-0.183 6, P=0.031), angiotensinⅡ (AngⅡ; Pearson's r=-0.218 6, P=0.010), ACE/ACE2 (Pearson's r=-0.319 2, P<0.001), AngⅡ/Ang1-7 (Pearson's r=-0.224 3, P=0.008), while the linear relationship with ACE2 and Ang1-7 was not significant (all P>0.05). 3) Basal follicle-stimulating hormone was positively correlated with age ( β=0.636, P<0.001), ACE2 ( β=0.267, P=0.026) and AngⅡ ( β=0.268, P=0.001), while negatively correlated with ACE ( β=-0.320, P<0.001) and Ang1-7 ( β=-0.217, P=0.014). Basal luteinizing hormone was positively correlated with AngⅡ ( β=0.330, P=0.003), while negative correlated with Ang1-7 ( β=-0.395, P=0.002). Antral follicle count was positively correlated with Ang1-7 ( β=0.153, P=0.049), while negatively correlated with age ( β=-0.869, P<0.001) and ACE2 ( β=-0.082, P=0.004). Basal anti-Müllerian hormone was only negatively correlated with age ( β=-0.349, P<0.001). There was no correlation between RAS and basal estradiol, progesterone, prolactin, testosterone (all P>0.05). 4) Oocyte retrieval count was positively correlated with renin ( β=0.146, P=0.014), AngⅡ ( β=0.113, P=0.034) and Ang1-7 ( β=0.185, P=0.002), while negatively correlated with age ( β=-0.717, P<0.001); MⅡ oocyte maturation rate was positively correlated with AngⅡ ( β=0.207, P=0.019) and Ang1-7 ( β=0.217, P=0.026), while negatively correlated with age ( β=-0.518, P<0.001). There was no correlation between RAS and the rates of two pronuclei embryos, transplantable embryos, high-quality embryos (all P>0.05). Conclusion:The local RAS in ovarian follicles is affected by age and correlated with ovarian function and IVF laboratory outcome.

Objective To investigate the relationship between gene polymorphisms of T cell immunoglobulin domain and mucin domain protein-3 (Tim-3) and ulcerative colitis (UC) in Han nationality of Zhejiang.Methods A total of 391 UC patients and 573 healthy controls were recruited.Two single nucleotide polymorphisms (SPNs) of Tim-3 (rs1036199 and rs10515746) were examined by the improved multiple ligase detection reaction technique.Chi-square test or Fisher's exact test was performed to analyze the differences in the distribution of Tim-3 gene polymorphisms and its influence on the location and severity.Haploview 4.2 software was used to analyze linkage disequilibrium (LD) and haplotype.Results The frequencies of genotype CA+AA and mutant allele A of rs10515746 in UC were lower than those in healthy controls (1.79％,7/391 vs 4.19％,24/573;0.90％,7/782 vs 2.18％,25/1 146;x2=4.295 and 4.712,P=0.038 and 0.030).However,there was no significant differences in frequencies of genotype CA+ CC and mutant allele C of gene rs1036199 between UC patients and the healthy controls (1.79％,7/891 vs 8.49％,20/578;0.90％,7/782 vs 1.74％,20/1 146;both P＞0.05).The frequencies of genotype CA+AA and mutant allele A of rs10515746 in mild and moderate UC patients were both higher than those in severe UC patients (2.87 ％,7/244 vs 0;1.43 ％,7/488 vs 0),and the differences were statistically significant (Fisher's exact test,P=0.049 and 0.048).The analysis for LD indicated that rs1036199andrs10515746 were closeLD (D'=0.92,r2=0.72).Furthermore,the frequency of haplotype CA formed by the mutant alleles C and A of these two SNPs was lower in UC patients than that in healthy controls (0.64％,5/782 vs 1.74％,20/1 146),and the difference was statistically significant (x2 =4.441,P=0.035).Conclusions Tim-3 (rs10515746) gene mutation may not only decrease the incidence,but also reduce the severity of UC.Moreover,the haplotype CA formed by the mutant alleles of rs1036199 and rs10515746 may also reduce the incidence of UC.

Objective To explore the relation between genetic polymorphisms and the expression in colonic tissues of solute-linked carrier family 26 member A3 (SLC26A3) and susceptibility of Crohn's disease (CD) in Han population of Zhejiang Province.Methods A total of 265 CD patients and 566 gender-and age-matched healthy individuals were enrolled.Alleles and genotypes of SLC26A3 (rs17154444,rs7810937,rs7785539,rs2108225,rs6951457) were examined by SNaPshot.The linkage disequilibrium (LD) and haplotype were also analyzed.Eight patients with colonic CD and eight genderand age-matched patients with benign colonic polyps (control group) were selected.The expression level of SLC26A3 protein in the colonic tissue was detected by immunohistochemistry.T test and rank-sum test were performed for statistical analysis.Unconditional Logistic regression analysis was used to analyze the distributions of SLC26A3 polymorphisms and their effects on the clinicopathological features of CD patients.Results The frequencies of mutant allele of rs2108225,rs7785539 and rs6951457 of the CD group were 53.77％ (285/530),4.72％ (25/530) and 2.83％ (15/530),and the frequencies of mutant genotype were 76.23 ％ (202/265),9.43 ％ (25/265) and 5.66 ％ (15/265),which were lower than those of the control group (60.95％,690/1 132;8.13％,92/1 132;6.10％,69/1 132;83.92％,475/566;15.37％,87/566 and 11.84％,67/566),and the differences were statistically significant (all P＜0.05).The frequencies of mutant allele of rs17154444 and rs7810937 of the CD group were 10.19％ (54/530) and 34.91 ％ (185/530),and the frequencies of mutant genotype were 18.49 ％ (49/265) and 56.23 ％ (149/ 265),compared with those of the control group (8.30％,94/1 132;30.92％,350/1 132;15.55％,88/566 and 51.77％,293/566),the differences were not statistically significant (all P＞0.05).The frequency of mutant allele G of rs2108225 in patients with ileal CD was 47.89 ％ (91/190),and the frequency of mutant genotypeAG+GG was 65.26％(62/95),which were both lower than those of colonic CD (61.62％,122/198 and 85.86％,85/99),and the differences were statistically significant (both P＜0.012 5).rs7810937,rs7785539 and rs2108225 were in a strong linkage disequilibrium.The frequencies of haplotypes AGG and ACA of the CD group were 53.96％ (286/530) and 4.34％ (23/530),which were lower than those of the control group (60.07％,680/1 132 and 7.51％,85/1 132),and the differences were statistically significant (52 =5.534,P=0.019;x2 =5.967,P=0.015).And the frequency of haplotype AGA of the CD group was 8.30％ (44/530),which was higher than that of the control group (1.15％,13/1 132),and the difference was statistically significant (x2 =7.793,P＜0.01).Furthermore,the expression level of SLC26A3 protein in colonic tissues of eight colonic CD patients was 0.19±0.07,which was lower than that of patients with benign colonic polyps (0.26 ±-0.03),and the difference was statistically significant (t=2.55,P=0.023).In addition,the expression levels of SLC26A3 protein in patients carrying genotype GG or AG of rs2108225 were 0.19±0.03 and 0.10±0.01,respectively,which were lower than that of patients carrying genotype AA (0.26± 0.02),and the differences were statistically significant (t=3.19,P=0.033;t=9.06,P=0.003).Conclusions The genetic polymorphismns and their haplotypes of SLC26A3 (rs7785539,rs2108225 and rs6951457) are associated with the susceptibility of CD,and SLC26A3 (rs2108225) polymorphism may affect the expression level of SLC26A3 protein in the colonic tissues.

Objective To investigate the application value of Photoshop in grading invasive risk of gastric stromal tumors (GSTs). Methods EUS image of 97 cases of GSTs confirmed by pathological and immunohistochemical examination were collected. GSTs were divided into four groups (very low risk, low risk, intermediate risk, high risk) by tumor size, mitotic count and rupture of tumor. Mean gray value (intensity of echo) and gray value standard deviation (uniformity of echo) of EUS images of the lesions were determined by Photoshop and then the differences of each group were found by statistical analysis. Results It is difficult to differentiate EUS images of GSTs from each group by visual observation. The mean gray value of EUS image of very low risk group,low risk group, intermediate risk group and high risk group of GSTs respectively were (56.54 ± 6.10), (59.20 ± 7.51), (77.77 ± 10.90) and (83.43 ± 12.47). There was no significant difference between very low risk group and low risk group (P > 0.05). There was no significant difference between intermediate risk group and high risk group (P > 0.05). In addition, the others all had significantly different from that of each group (P < 0.05). The mean gray value standard deviation of EUS image of very low risk group, low risk group, intermediate risk group and high risk group of GSTs respectively were (8.46 ± 2.59), (12.57 ± 5.89), (12.84 ± 4.15) and (16.69 ± 4.69). There was no significant difference between low risk group and intermediate risk group (P > 0.05). In addition, the others all had significantly different from that of each group (P < 0.05). Conclusions The higher risk of GSTs, the higher of echo intensity and the worse of echo uniformity under EUS. Photoshop combined with EUS is helpful for differentiating different risk of GSTs by analyzing mean gray value and gray value standard deviation of the lesions.