Background/Aims: Liver cirrhosis involves chronic inflammation and progressive fibrosis.Among various immune cells, CD8+ T cells are considered a major contributor to hepatic inflammation and fibrosis. However, the exact molecular pathways governing CD8+ T-cell-mediated effects in cirrhosis remain unclear. Methods: This study analyzed transcriptomic and single-cell sequencing data to elucidate CD8+ T-cell heterogeneity and implications in cirrhosis. Results: Weighted gene co-expression analysis of bulk RNA-seq data revealed an association between cirrhosis severity and activated T-cell markers like HLA and chemokine genes. Furthermore, single-cell profiling uncovered eight CD8+ T-cell subtypes, notably, effector memory (Tem) and exhausted (Tex) T cells. Tex cells, defined by PDCD1, LAG3, and CXCL13 expression, were increased in cirrhosis, while Tem cells were decreased. Lineage tracing and differential analysis highlighted CXCL13+ Tex cells as a terminal, exhausted subtype of cells with roles in PD-1 signaling, glycolysis, and T-cell regulation. CXCL13+ Tex cells displayed T-cell exhaustion markers like PDCD1, HAVCR2, TIGIT, and TNFRSF9. Functional analysis implicated potential roles of these cells in immunosuppression. Finally, a CXCL13+ Tex-cell gene signature was found that correlated with cirrhosis severity and poorer prognosis of liver cancer. Conclusions In summary, this comprehensive study defines specialized CD8+ T-cell subpopulations in cirrhosis, with CXCL13+ Tex cells displaying an exhausted phenotype associated with immune dysregulation and advanced disease. Key genes and pathways regulating these cells present potential therapeutic targets.

Background/Aims: Liver cirrhosis involves chronic inflammation and progressive fibrosis.Among various immune cells, CD8+ T cells are considered a major contributor to hepatic inflammation and fibrosis. However, the exact molecular pathways governing CD8+ T-cell-mediated effects in cirrhosis remain unclear. Methods: This study analyzed transcriptomic and single-cell sequencing data to elucidate CD8+ T-cell heterogeneity and implications in cirrhosis. Results: Weighted gene co-expression analysis of bulk RNA-seq data revealed an association between cirrhosis severity and activated T-cell markers like HLA and chemokine genes. Furthermore, single-cell profiling uncovered eight CD8+ T-cell subtypes, notably, effector memory (Tem) and exhausted (Tex) T cells. Tex cells, defined by PDCD1, LAG3, and CXCL13 expression, were increased in cirrhosis, while Tem cells were decreased. Lineage tracing and differential analysis highlighted CXCL13+ Tex cells as a terminal, exhausted subtype of cells with roles in PD-1 signaling, glycolysis, and T-cell regulation. CXCL13+ Tex cells displayed T-cell exhaustion markers like PDCD1, HAVCR2, TIGIT, and TNFRSF9. Functional analysis implicated potential roles of these cells in immunosuppression. Finally, a CXCL13+ Tex-cell gene signature was found that correlated with cirrhosis severity and poorer prognosis of liver cancer. Conclusions In summary, this comprehensive study defines specialized CD8+ T-cell subpopulations in cirrhosis, with CXCL13+ Tex cells displaying an exhausted phenotype associated with immune dysregulation and advanced disease. Key genes and pathways regulating these cells present potential therapeutic targets.

Background/Aims: Liver cirrhosis involves chronic inflammation and progressive fibrosis.Among various immune cells, CD8+ T cells are considered a major contributor to hepatic inflammation and fibrosis. However, the exact molecular pathways governing CD8+ T-cell-mediated effects in cirrhosis remain unclear. Methods: This study analyzed transcriptomic and single-cell sequencing data to elucidate CD8+ T-cell heterogeneity and implications in cirrhosis. Results: Weighted gene co-expression analysis of bulk RNA-seq data revealed an association between cirrhosis severity and activated T-cell markers like HLA and chemokine genes. Furthermore, single-cell profiling uncovered eight CD8+ T-cell subtypes, notably, effector memory (Tem) and exhausted (Tex) T cells. Tex cells, defined by PDCD1, LAG3, and CXCL13 expression, were increased in cirrhosis, while Tem cells were decreased. Lineage tracing and differential analysis highlighted CXCL13+ Tex cells as a terminal, exhausted subtype of cells with roles in PD-1 signaling, glycolysis, and T-cell regulation. CXCL13+ Tex cells displayed T-cell exhaustion markers like PDCD1, HAVCR2, TIGIT, and TNFRSF9. Functional analysis implicated potential roles of these cells in immunosuppression. Finally, a CXCL13+ Tex-cell gene signature was found that correlated with cirrhosis severity and poorer prognosis of liver cancer. Conclusions In summary, this comprehensive study defines specialized CD8+ T-cell subpopulations in cirrhosis, with CXCL13+ Tex cells displaying an exhausted phenotype associated with immune dysregulation and advanced disease. Key genes and pathways regulating these cells present potential therapeutic targets.

Background/Aims: Liver cirrhosis involves chronic inflammation and progressive fibrosis.Among various immune cells, CD8+ T cells are considered a major contributor to hepatic inflammation and fibrosis. However, the exact molecular pathways governing CD8+ T-cell-mediated effects in cirrhosis remain unclear. Methods: This study analyzed transcriptomic and single-cell sequencing data to elucidate CD8+ T-cell heterogeneity and implications in cirrhosis. Results: Weighted gene co-expression analysis of bulk RNA-seq data revealed an association between cirrhosis severity and activated T-cell markers like HLA and chemokine genes. Furthermore, single-cell profiling uncovered eight CD8+ T-cell subtypes, notably, effector memory (Tem) and exhausted (Tex) T cells. Tex cells, defined by PDCD1, LAG3, and CXCL13 expression, were increased in cirrhosis, while Tem cells were decreased. Lineage tracing and differential analysis highlighted CXCL13+ Tex cells as a terminal, exhausted subtype of cells with roles in PD-1 signaling, glycolysis, and T-cell regulation. CXCL13+ Tex cells displayed T-cell exhaustion markers like PDCD1, HAVCR2, TIGIT, and TNFRSF9. Functional analysis implicated potential roles of these cells in immunosuppression. Finally, a CXCL13+ Tex-cell gene signature was found that correlated with cirrhosis severity and poorer prognosis of liver cancer. Conclusions In summary, this comprehensive study defines specialized CD8+ T-cell subpopulations in cirrhosis, with CXCL13+ Tex cells displaying an exhausted phenotype associated with immune dysregulation and advanced disease. Key genes and pathways regulating these cells present potential therapeutic targets.

Objective:To study the sex-related growth difference (SRGD) of embryo morphological parameters and early serum β-human chorionic gonadotropin (β-hCG) level in single embryo transfer (SET) singleton live birth cycles.Methods:A retrospective cohort study analyzed the clinical data of patients with fresh and frozen non-donor, SET, and singleton live birth cycles from January 2020 to December 2022 in the Reproductive Medicine Center of Yantaishan Hospital ( n=311), and the neonatal sex group was grouped by gender, with 154 cases in the male infant group and 157 cases in the female infant group. The embryo transfer stage (93 cases in cleavage stage, 218 cases in blastocysts) and embryo quality (250 cases in the good-quality embryo group and 61 cases in the non-good-quality embryo group) were analyzed by subgroup stratification. Univariate and binary logistic/stepwise regression analysis were used to analyze whether there were embryonic morphological parameters and early β-hCG levels in the embryonic development stage and early pregnancy. Results:Univariate analysis showed that the general data matching of patients, such as parents' age, body mass index, embryo transfer stage, embryo quality, insemination method, and cycle type were comparable (all P>0.05). Univariate analysis of sex-related morphological parameters showed that the number of cells on the day 2 (D2) embryo in the male infant group (4.25±0.94) was larger than that in the female infant group (3.98±0.84, P=0.007). Binary logistic regression analysis showed that the number of cells on D2 embryo was positively correlated with the number of live-born males ( OR=1.428, 95% CI: 1.052-1.938, P=0.022). Subgroup analysis of blastocysts showed that the number of cells on D2 embryo ( OR=1.522, P=0.021) and blastocyst expansion ( OR=2.969, P=0.029) were positively correlated with live-born males. The early β-hCG level in the male infant group [776.40 (521.95, 1 127.25) U/L] was higher than that in the female infant group [634.60 (425.80, 973.05) U/L, P=0.003], and it was related to the embryo transfer period ( β=0.139, P=0.012) and embryo quality ( β=0.136, P=0.014). In the stratified analysis of different embryo stages and quality subgroups, β-hCG levels in male infant group during the cleavage stage, blastocyst stage, and high-quality embryos [677.20 (462.63, 1 028.50) U/L, 838.30 (557.50, 1 191.00) U/L, 816.00 (563.95, 1 199.75) U/L] were higher than those in female infant group [538.40 (344.80, 804.80) U/L, 724.95 (446.83, 1 016.75) U/L, 651.40 (431.30, 985.73) U/L], and the differences were statistically significant ( P=0.041, P=0.044, P=0.001). Subgroup regression analysis of blastocysts showed that β-hCG level was positively correlated with higher blastocyst expansion ( β=0.162, P=0.010) and higher trophectoderm grade ( β=0.344, P<0.001). Conclusion:In the single live birth cycle of SET, the number of D2 embryo cells and the early β-hCG level of male infants were higher than those of female infants, and the expansion degree of blastocyst is related to sex, and there is SRGD.

Objective:To study the sex-related growth difference (SRGD) of embryo morphological parameters and early serum β-human chorionic gonadotropin (β-hCG) level in single embryo transfer (SET) singleton live birth cycles.Methods:A retrospective cohort study analyzed the clinical data of patients with fresh and frozen non-donor, SET, and singleton live birth cycles from January 2020 to December 2022 in the Reproductive Medicine Center of Yantaishan Hospital ( n=311), and the neonatal sex group was grouped by gender, with 154 cases in the male infant group and 157 cases in the female infant group. The embryo transfer stage (93 cases in cleavage stage, 218 cases in blastocysts) and embryo quality (250 cases in the good-quality embryo group and 61 cases in the non-good-quality embryo group) were analyzed by subgroup stratification. Univariate and binary logistic/stepwise regression analysis were used to analyze whether there were embryonic morphological parameters and early β-hCG levels in the embryonic development stage and early pregnancy. Results:Univariate analysis showed that the general data matching of patients, such as parents' age, body mass index, embryo transfer stage, embryo quality, insemination method, and cycle type were comparable (all P>0.05). Univariate analysis of sex-related morphological parameters showed that the number of cells on the day 2 (D2) embryo in the male infant group (4.25±0.94) was larger than that in the female infant group (3.98±0.84, P=0.007). Binary logistic regression analysis showed that the number of cells on D2 embryo was positively correlated with the number of live-born males ( OR=1.428, 95% CI: 1.052-1.938, P=0.022). Subgroup analysis of blastocysts showed that the number of cells on D2 embryo ( OR=1.522, P=0.021) and blastocyst expansion ( OR=2.969, P=0.029) were positively correlated with live-born males. The early β-hCG level in the male infant group [776.40 (521.95, 1 127.25) U/L] was higher than that in the female infant group [634.60 (425.80, 973.05) U/L, P=0.003], and it was related to the embryo transfer period ( β=0.139, P=0.012) and embryo quality ( β=0.136, P=0.014). In the stratified analysis of different embryo stages and quality subgroups, β-hCG levels in male infant group during the cleavage stage, blastocyst stage, and high-quality embryos [677.20 (462.63, 1 028.50) U/L, 838.30 (557.50, 1 191.00) U/L, 816.00 (563.95, 1 199.75) U/L] were higher than those in female infant group [538.40 (344.80, 804.80) U/L, 724.95 (446.83, 1 016.75) U/L, 651.40 (431.30, 985.73) U/L], and the differences were statistically significant ( P=0.041, P=0.044, P=0.001). Subgroup regression analysis of blastocysts showed that β-hCG level was positively correlated with higher blastocyst expansion ( β=0.162, P=0.010) and higher trophectoderm grade ( β=0.344, P<0.001). Conclusion:In the single live birth cycle of SET, the number of D2 embryo cells and the early β-hCG level of male infants were higher than those of female infants, and the expansion degree of blastocyst is related to sex, and there is SRGD.

Objective:To investigate the correlations between expression of long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), nuclear-enriched abundant transcript 1 (NEAT1) and their functions on exosome secretion, proliferation and invasion in hepatocellular carcinoma (HCC).Methods:We used small interfering RNA of MALAT1 (si-MALAT1) to knockdown MALAT1 in HuH-7. At the meanwhile, cells which were transfected with si-NC were used as the negative control group. Expression of NEAT1, cell proliferation and invasion function were detected these two groups. HuH-7 cells were transfected with lentivirus NEAT1 over expressing vector (lv-NEAT1) or negative control (lv-control). Expression of exosomes secretion related genes were analyzed between lv-NEAT1 and lv-control groups. Cells of lv-NEAT1 were knockdown MALAT1 expression using si-MALAT1, which could be si-MALAT1+ lv-NEAT1 group. exosomes secretion was detected in si-NC, si-MALAT1 and si-MALAT1+ lv-NEAT1 group. We treated cells (si-MALAT1 group) with exosomes from cells with lv-NEAT1 or lv-control to divide cells as si-MALAT1+ exosomes of lv-NEAT1 cells and si-MALAT1+ exosomes of lv-control groups. Cell proliferation and invasion of cells were detected in two groups.Results:Low expression of NEAT1 were found in MALAT1 knockdown cells compared with si-NC group [(0.72±0.02) vs. (0.98±0.01), P<0.05]. Cells with MALAT1 knockdown shown diminished proliferation [(0.66±0.03) vs. (0.98±0.04), P<0.05)] and invasion [(88.33±7.26) vs. (147.70±13.62), P<0.05)]. Compared with si-NC group, CD9 and CD63 expression were decreased in exosomes of si-MALAT1 group. Compared with si-MALAT1 group, CD9 and CD63 expression was increased in exosomes of si-MALAT1+ lv-NEAT1 group. Compared with si-MALAT1+ exosomes of lv-control group, proliferation [(0.97±0.03) vs. (0.74±0.05), P<0.05)] and invasion [ (132.70±7.36) vs. (98.33±6.01), P<0.05) ] were increased in si-MALAT1+ exosomes of lv-NEAT1 group. Exosomes related genes expression including HSPA8 (5.53±0.31), SLC3A2 (0.32±0.07) and SLC7A5 (0.77±0.45) were changed in lv-NEAT1 group compared with lv-control group [(0.98±0.15), P<0.05]. Conclusion:MALAT1 induced exosomes secretion by NEAT1 and exosomes related genes regulation. This regulation might be related with increased proliferation and invasion function in HCC cells with MALAT1 and NEAT1 abnormal expression.

Objective:To investigate the related risk factors for biliary anastomotic stenosis after liver transplantation (LT) from donation after cardiac death(DCD) and therapeutic strategies.Methods:The data of 192 patients who received LT from DCD in First Hospital of Kunming from Jan 2010 to Jun 2018 were retrospectively analyzed. A total of 145 patients were enrolled, 85 males and 60 females, with average age 45 years. There was a biliary anastomotic stenosis in 8 cases and no stenosis in 137 cases. Their Chinese criterion for biliary anatomic stenosis, age, body mass index, liver fat, cold/warm ischemia time, unschedule cardiac arrest time, usage of vasopressors, high sodium in the donor were compared, and stenosis related factors were analysed by Spearman correlation analysis.Results:The stenosis was positively correlated with age ( r=0.229), body mass index ( r=0.204), lipoidosis ( r=0.239), duration of hot ischemia ( r=0.214), total duration of unplanned cardiac arrest ( r=0.401), use of booster drugs ( r=0.237), and preoperative donor hypernatremia ( r=0.557) (all P<0.05). Endoscopic biliary stent implantation is effective in the treatment of biliary anastomotic stenosis and has a high success rate. Conclusions:There are many factors related to biliary anastomotic stenosis after DCD liver transplantation, but the better donor maintenance, shorten cold/ warm ischemia time, improved anastomosis will be helpful to reduce biliary complications.As the same time, endoscopic biliary stent placement is the preferred way to treat biliary anastomotic stenosis.

Objective: To explore the feasibility, safety and long-term efficacy of laparoscopic total gastrectomy combined with distal pancreaticosplenectomy for the treatment of T4b gastric cancer. Methods: A retrospective cohort study was performed. Clinical data of consecutive patients with T4b gastric cancer invading pancreatic tail undergoing laparoscopic or open total gastrectomy combined with distal pancreaticosplenectomy from January 2010 to December 2014 were analyzed retrospectively. Enrollment criteria: (1) primary gastric cancer confirmed by pathology as T4b adenocarcinoma; (2) chest+abdominal+pelvic enhanced CT indicated cancer invading pancreatic tail without distant metastasis, and R0 resection was evaluated as feasible before operation; (3) physical status was ECOG score 0 to 2, and was tolerant to operation. Patients with peritoneal implant metastasis and tumor invasion of other organs during operation, or changes in surgical methods for other reasons were excluded. All the operations were performed by the same surgical team, which had the experiences of more than 100 cases of laparoscopic and 100 cases of open radical gastrectomy with D2 lymph node dissection. The choice of surgical procedure was discussed by the surgeon and the patient, and decided according to the patient′s intension. Patients were divided into the laparoscopic group and open group according to the surgical method. Intraoperative and perioperative findings were compared between the two groups. The 3-year disease-free survival rate were analyzed with Kaplan-Meier survival curve and compared by using log-rank test. Results: A total of 37 consecutive patients were enrolled, including 21 in the laparoscopic group and 16 in the open group, and no one receiving laparoscopic procedure was converted to open surgery. The baseline data of two groups were comparable (all P>0.05). Compared with the open group, the laparoscopic group had significantly longer operation time [(264.0±35.1) minutes vs. (226.6±49.9) minutes, t=2.685, P=0.011], significantly less intraoperative blood loss [(65.7±37.4) ml vs. (182.2±94.6) ml, t=-4.658, P<0.001], significantly shorter time to postoperative flatus [(2.8±0.7) days vs. (4.1±0.7) days, t=-5.776, P<0.001] and significantly shorter postoperative hospital stay [(13.3±2.8) days vs. (16.6±4.3) days, t=-2.822, P=0.008]. Morbidity of postoperative complications, including anastomotic leakage, pancreatic fistula, abdominal abscess, intraperitoneal hemorrhage and duodenal stump leakage, in two groups was similar [19.0% (4/21) vs. 4/16, P=0.705]. There were no cases of anastomotic bleeding or stenosis. The 30-day postoperative mortality was 0 in the laparoscopic group and 1/16 in the open group, respectively (P=0.432). The 3-year disease-free survival rates were 38.1% and 37.5% in the laparoscopic and open group, respectively (P=0.751). Conclusion Laparoscopic total gastrectomy combined with distal pancreaticosplenectomy performed by experienced surgeons for T4b gastric cancer is safe and effective.

Objective To evaluate GIPR protein expression in predicting prognosis of appendical neuroendocrine tumors (A-NET) patients.Methods The clinical data of 40 A-NET cases surgically treated from June 2007 to June 2017 at Tianjin Police Hospital were analyzed.The expression of GIPR markers was detected by immunohistochemistry (IHC).Results There were 25 male and 15 female patients,with an median age of 59 years.Sex,tumor site,tumor size,tumor stage and lymph node metastasis were not related to prognosis (P ＞ 0.05).The expression of GIPR was positive in 18 cases and negative in 22 cases.There were 16 cases in G1 stage,20 cases in G2,4 cases in G3.The expression of GIPR protein and pathological grades were related to prognosis (P ＜ 0.05).Conclusions Symptoms of appendix neuroendocrine tumor are nonspecific.Diagnosis is dependent on pathological examination and immunohistochemistry.Positive GIPR protein expression predicts poor prognosis for A-NETs patients.