BACKGROUND:Superovulation is a common therapy in assisted reproductive technology.In clinical practice,some patients experience repeated superovulation to get pregnant. OBJECTIVE:To explore the effect of repeated superovulation on the developmental potential of oocytes in mice and humans. METHODS:Both animal experiments and retrospective clinical research were conducted.The animal study involved 90 SPF grade ICR 8-week-old female mice,who were randomly divided into three groups for 1,3,and 5 superovulations,respectively.The clinical study involved 306 patients who had undergone three consecutive in vitro fertilization cycles.The number of ovules obtained and embryonic development in different cycles were compared. RESULTS AND CONCLUSION:(1)The animal study indicated that repeated superovulation did not affect the embryonic development or developmental speed of mouse embryos.Similarly,there was no significant difference in the mouse blastocyst apoptosis,DNA damage,or the formation of inner cell mass and trophectoderm(P>0.05).(2)The clinical study also revealed no significant differences in the number of retrieved oocytes(8.60±5.04,8.58±4.87,and 8.38±4.63,P=0.81)and transferable embryos(2.42±1.99,2.40±1.92,and 2.64±2.00,P=0.26)over the three cycles.(3)In both the young group(<35 years)and the old group(≥35 years),the embryo quality was not affected by repeated superovulation(P>0.05).(4)These findings show that repeated superovulation does not affect the developmental potential of oocytes in mice and humans.

Objective To compare the short- to mid-term outcomes of aortic valve plasty (AVP) and Ross surgery in children with severe aortic valve disease. Methods The patients (aged<18 years) with severe aortic valve disease who underwent AVP (an AVP group) or Ross surgery (a Ross group) at the Department of Cardiovascular Surgery, West China Hospital from January 2019 to September 2023 were retrospectively included. We compared perioperative and follow-up data between the groups. Results A total of 48 pediatric patients were included, including 28 males and 20 females, with an average age of (9.3±4.5) years. There were 25 patients in the AVP group, and 23 in the Ross group. Leaflet thinning (15/25, 60.0%) and leaflet extension (10/25, 40.0%) were the most common strategies used in the AVP group, while root replacement technique (12/23, 52.2%) and subcoronary technique (10/23, 43.5%) were the most frequently used strategies in the Ross group. There was no in-hospital death. The median follow-up time was 16.0 (7.0, 30.0) months. Peak flow velocity of the aortic valve was higher in the AVP group [2.0 (1.4, 2.9) m/s vs. 1.2 (1.0, 1.5) m/s, P<0.001], while there was no statistical difference in the postoperative aortic valve regurgitation severity between the two groups (P=0.127). During follow-up, the overall reoperation rate and aortic valve reoperation rate were similar between the AVP group and the Ross group (8.0% vs. 13.0%, P=0.922; 8.0% vs. 0.0%, P=0.266). The rate of recurrent aortic valve disease was higher in the AVP group (52.0% vs. 4.3%, P<0.001), while further analysis failed to recognize any risk factors. Conclusion AVP and Ross procedure show similar perioperative safety, survival and reoperation rate. The rate of recurrent aortic valve disease is higher in the AVP group, but further investigations are needed to confirm the causes.

Objective:To investigate the effects and mechanism of Jinlong Bushen Mixture against lipid metabolism disorders induced by high-sugar and high-fat diet in rats with metabolic syndrome by combining network pharmacology and experimental validation.Methods:TCMSP and related literature were retrieved to obtain the active components and targets of wolfberry, golden cherry, longan meat, jujube, gynostemma, rosmarinus officinalis, and motherwort in Jinlong Bushen Mixture, and GeneCards online database was retrieved to obtain metabolic syndrome-related targets. Venny 2.1.0 was used to obtain the intersection targets of Jinlong Bushen Mixture and metabolic syndrome, and STRING online STRING online database was used to construct the PPI network of intersecting targets. Core targets in the top 50 degree values were screened using the Cyto NCA plugin in Cytoscape 3.9.0. The DAVID Bioinformatics Resources 6.8 online analysis platform was used for GO functional enrichment and KEGG pathway enrichment analysis. Metabolic syndrome rat model was established using high sugar, high salt, and high fat feed for 20 weeks. The successfully modeling rats were divided into model groups, positive control group and Jinlong Bushen Mixture group according to random number table, and a blank control group was also set up, with 8 rats in each group. Jinlong Bushen Mixture group was gavaged with Jinlong Bushen Mixture 1.8 ml/kg, a positive control group was gavaged with Metformin 90 mg/kg, and the blank and model groups were gavaged with an equal volume of saline, 1 time/d, for 4 weeks. The changes in body weight, abdominal circumference, and fasting blood glucose in rats were observed. The blood lipid level in rats was detected. The pathological changes of liver tissues were detected by HE staining. The levels of ATP, IL-1β, and IL-6 in liver tissues were determined by ELISA. The mRNA expression of liver kinase B1 (LKB1), AMPK, Akt1, carnitine palmitoyltransferase 1A (CPT1A), and downregulated lactate dehydrogenase A (LDH-A) in liver tissue were detected by qRT-PCR. Western blot was used to determine the expression of p-LKB1, LKB1, p-AMPK, AMPK, p-Akt1, and Akt1 in liver tissue were detected by Western blotting.Results:A total of 141 main active components, 841 active targets, 18 763 metabolic syndrome targets, and 820 drug-disease intersection targets of Jinlong Bushen Mixture were obtained. The KEGG pathway enrichment analysis yielded 173 entries, including mainly the PI3K-Akt, and HIF-1 signalling pathway. The experimental results showed that the weight, fasting blood glucose, and lipid levels of rats in the Jinlong Bushen mixture group decreased, and the disorder of liver glucose and lipid metabolism in rats improved; the levels of ATP, IL-1β and IL-6 decreased ( P<0.05); The mRNA expression of LKB1, AMPK, Akt1, and CPT1A in liver tissue increased ( P<0.05), while LDH-A mRNA expression decreased ( P<0.05). The p-LKB1/LKB1, p-AMPK/AMPK, and p-Akt1/Akt1 ratio increased ( P<0.05). Conclusion:Jinlong Bushen Mixture may restore lipid metabolism disorders in the metabolic syndrome through multiple targets and pathways. Its mechanism may exert pharmacological effects through the LKB1/AMPK/Akt signaling pathway.

Objective:To compare the anesthetic potency of dexmedetomidine combined with remifentanil for colonoscopy in the patients with different body mass indexes (BMIs) to assess the clinical significance of the influence of weight on the level of pain during the procedure.Methods:American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ patients, aged 18-64 yr, undergoing elective colonoscopy, were divided into 3 groups based on the BMI: group Ⅰ (underweight group, BMI<18.5 kg/m 2), group Ⅱ (normal weight group, BMI 18.5-24.0 kg/m 2), and group III (overweight group, 24.0 kg/m 2 < BMI <30.0 kg/m 2). The prescribed dose of dexmedetomidine was infused within 2 min, then remifentanil was infused as a bolus of 1 μg/kg within 2 min followed by an infusion of 0.1 μg · kg -1 · min -1 throughout the surgery, and then colonoscopy was performed in patients of each group. The up-and-down sequential allocation was used to determine the dose of dexmedetomidine, the initial dose of dexmedetomidine in each group was 0.3 μg/kg, and the ratio between the two successive doses was 1.2. The positive response was defined as the Modified Observer′s Assessment of Alertness/Sedation Scale score > 1 and occurrence of body movement during the operation. Each time the dose of dexmedetomidine increased/decreased in the next patient depending on whether or not the response was positive. The median effective dose (ED 50) and 95% confidence interval ( CI) of dexmedetomidine were calculated using the Dixon-Massey formula. Results:Compared with group Ⅰ (0.42 [95% CI 0.38-0.47] μg/kg), the ED 50 of dexmedetomidine was significantly decreased in group II (0.23 [95% CI 0.19-0.32] μg/kg) and in group III (0.18 [95% CI 0.13-0.22] μg/kg) ( P<0.05). The ED 50 of dexmedetomidine was significantly decreased in group Ⅲ when compared with group Ⅱ ( P<0.05). Conclusions:With the increase of patients′ BMIs, the anesthetic potency of dexmedetomidine for colonoscopy is significantly enhanced when combined with remifentanil, indicating that clinicians should pay attention to the influence of weight on the level of pain during procedures.

Objective:To evaluate the effect of sleep fragmentation on postoperative cognitive dysfunction (POCD) and hippocampal glutaminergic metabolism in aged mice anesthetized with isoflurane.Methods:Forty healthy SPF-grade male C57BL/6J mice, aged 18 months, weighing 20-30 g, were divided into 4 groups ( n= 10 each) by the random number table method: normal control group (group C), sleep fragmentation group (group SF), isoflurane anesthesia/surgery group (group I/S), and sleep fragmentation plus isoflurane anesthesia/surgery group (group SF+ I/S). Group C did not received any treatment. Group SF received sleep fragmentation for 24 h. The right carotid artery exposure was performed under isoflurane anesthesia in group I/S. Group SF+ I/S received isoflurane anesthesia/right carotid artery exposure at 24 h after sleep fragmentation. The metabolic levels of glutamate (Glu), glutamine (Gln), Glu/Gln complex (Glx), and N-acetylaspartate (NAA) and their ratio to creatine (Cr) were measured by in vivo 9.4T hydrogen proton magnetic resonance spectroscopy at 2 h after anaesthesia. Y maze and Morris water maze tests were used to evaluate the cognitive function at 1-7 days after surgery. The mice were sacrificed after the behavioral testing, brain tissues were immediately obtained, and the number of Nissl bodies and density of dendritic spines in the hippocampal CA1 region were measured by Nissl staining and Golgi staining, respectively. Results:Compared with group C, the percentage of exploration time and shuttle times at the novel arm were significantly decreased, the number of crossing the original platform was decreased, the time of stay at the target quadrant was shortened, the ratios of Glu/Cr, Gln/Cr and Glx/Cr in the hippocampal CA1 region were increased, and the ratio of NAA/Cr was decreased, and the number of Nissl bodies and density of dendritic spines were decreased in SF, I/S and SF+ I/S groups ( P<0.05). Compared with group SF and group I/S, the percentage of exploration time and shuttle times at the novel arm were significantly decreased, the number of crossing the original platform was decreased, the time of stay at the target quadrant was shortened, the ratios of Glu/Cr and Glx/Cr in hippocampal CA1 region was increased, the ratio of NAA/Cr was decreased, and the number of Nissl bodies and density of dendritic spines were decreased in group SF+ I/S ( P<0.05). Conclusions:Sleep fragmentation exacerbates POCD in aged mice anesthetized with isoflurane, and the mechanism is related to nerve injury induced by abnormality in hippocampal glutaminergic metabolism excitability.

Objective:To investigate the self-healing of congenital auricular deformity (CAD) and the efficacy of the auricle corrector in infants.Methods:The infants with CAD who were born in the Affiliated Maternity and Child Health Care Hospital of Nantong University from January to December 2020 were collected. The patients who disagreed with the use of auricle corrector were included in the observation group. The patients were followed up on the day of one month and one year after birth by phone or WeChat, and the self-healing rate (the proportion of the number of ears found to be healed in the total number of affected ears after follow-up) of auricle was calculated. The patients treated with auricle corrector were included in the non-invasive correction group, which was treated with domestic auricle corrector. And according to the initial correction age, the patients were divided into ≤7 d, 8-28 d and >28 d three subgroups. The patients were followed up on the day of one month and one year after treatment by phone or WeChat, and the effective rate(the proportion of healed ears to the total number of affected ears after treatment) was calculated. And statistical analysis was used in the three subgroups of initial correction age ≤7 d, 8-28 d, >28 d. Variance analysis was used for general comparison between three groups, and LSD- t test for the comparison between two groups. P<0.05 was considered statistically significant. Results:A total of 43 children (71 ears, 34 left ears and 37 right ears) were included in the observation group, including 21 males and 22 females, (2.1±0.3) days after birth. In the non-invasive correction group, 24 children (41 ears, 20 left ears and 21 right ears) were included, including 8 males and 16 females, (29.1±23.8) days after birth. There were 6 cases (10 ears) in the initial correction age group ≤7 days, 6 cases (9 ears) in the 8-28 days group, and 12 cases (22 ears) in the >28 days group. In the observation group (71 ears), the rates of self-healing on the day of one month and one year after birth were 29/71 and 30/71, respectively, including 23/31 and 24/31 for lop ear, 3/6 for Stahl’s ear, 3/3 for complex deformity, and 0 for other ear deformities. The effective rates were 41/41 and 35/41 one month and one year after treatment in the 24 children (41 ears) in the non-invasive correction group, respectively. Among them, the helical rim deformity was 12/12, lop ear 3/3, cup ear 7/7 and 5/7, Stahl’s ear 4/4, contracted ear 6/6, 5/6, prominent ear 2/2 and 0, the complex deformity 6/6 and 5/6, and microtia(Ⅰ) 1/1. The auricular deformities reoccurred in a total of 4 children (6 ears) 1 year after the end of treatment. According to the initial correction age into three groups, the treatment duration of the auricle corrector was (31.8±11.2) days, (31.3±10.1) days and (41.8±13.8) days, and the difference among the three groups was statistically significant ( F=3.42, P=0.043). There was no statistical significance between the groups ≤7 days and 8-28 days ( P=0.936), but there was statistical significance in the treatment duration between the groups ≤7 days and >28 days ( P=0.043), and between the groups 8-28 days and > 28 days ( P=0.041). Conclusion:CAD can self-heal to some extent. Self-healing rate is higher in lop ear and Stahl’s ears. The domestic auricle corrector has a good effect on CAD. The older the age of initial correction is, the longer the time of correction is. The type of CAD has a certain influence on the correction effect.

Objective:To investigate the self-healing of congenital auricular deformity (CAD) and the efficacy of the auricle corrector in infants.Methods:The infants with CAD who were born in the Affiliated Maternity and Child Health Care Hospital of Nantong University from January to December 2020 were collected. The patients who disagreed with the use of auricle corrector were included in the observation group. The patients were followed up on the day of one month and one year after birth by phone or WeChat, and the self-healing rate (the proportion of the number of ears found to be healed in the total number of affected ears after follow-up) of auricle was calculated. The patients treated with auricle corrector were included in the non-invasive correction group, which was treated with domestic auricle corrector. And according to the initial correction age, the patients were divided into ≤7 d, 8-28 d and >28 d three subgroups. The patients were followed up on the day of one month and one year after treatment by phone or WeChat, and the effective rate(the proportion of healed ears to the total number of affected ears after treatment) was calculated. And statistical analysis was used in the three subgroups of initial correction age ≤7 d, 8-28 d, >28 d. Variance analysis was used for general comparison between three groups, and LSD- t test for the comparison between two groups. P<0.05 was considered statistically significant. Results:A total of 43 children (71 ears, 34 left ears and 37 right ears) were included in the observation group, including 21 males and 22 females, (2.1±0.3) days after birth. In the non-invasive correction group, 24 children (41 ears, 20 left ears and 21 right ears) were included, including 8 males and 16 females, (29.1±23.8) days after birth. There were 6 cases (10 ears) in the initial correction age group ≤7 days, 6 cases (9 ears) in the 8-28 days group, and 12 cases (22 ears) in the >28 days group. In the observation group (71 ears), the rates of self-healing on the day of one month and one year after birth were 29/71 and 30/71, respectively, including 23/31 and 24/31 for lop ear, 3/6 for Stahl’s ear, 3/3 for complex deformity, and 0 for other ear deformities. The effective rates were 41/41 and 35/41 one month and one year after treatment in the 24 children (41 ears) in the non-invasive correction group, respectively. Among them, the helical rim deformity was 12/12, lop ear 3/3, cup ear 7/7 and 5/7, Stahl’s ear 4/4, contracted ear 6/6, 5/6, prominent ear 2/2 and 0, the complex deformity 6/6 and 5/6, and microtia(Ⅰ) 1/1. The auricular deformities reoccurred in a total of 4 children (6 ears) 1 year after the end of treatment. According to the initial correction age into three groups, the treatment duration of the auricle corrector was (31.8±11.2) days, (31.3±10.1) days and (41.8±13.8) days, and the difference among the three groups was statistically significant ( F=3.42, P=0.043). There was no statistical significance between the groups ≤7 days and 8-28 days ( P=0.936), but there was statistical significance in the treatment duration between the groups ≤7 days and >28 days ( P=0.043), and between the groups 8-28 days and > 28 days ( P=0.041). Conclusion:CAD can self-heal to some extent. Self-healing rate is higher in lop ear and Stahl’s ears. The domestic auricle corrector has a good effect on CAD. The older the age of initial correction is, the longer the time of correction is. The type of CAD has a certain influence on the correction effect.

Objective:To investigate the patients with hepatocellular carcinoma suitable for transcatheter arterial chemoembolization (TACE) after radical resection who were screened based on microvascular invasion (MVI) and Ki-67 expression.Methods:Of 400 patients with hepatocellular carcinoma who underwent radical resection in the Affiliated Hospital of Qingdao University from January 2013 to December 2019 were included and analyzed retrospectively, including 324 males and 76 females, aged (59.7±9.8) years, ranging from 32 to 87 years. According to whether they received adjuvant TACE treatment after operation, they were divided into simple operation group ( n=210) and TACE + operation group ( n=190). The recurrence in the first year after operation was followed up by outpatient reexamination. Univariate and multivariate Cox regression analysis were used to analyze the influencing factors of recurrence free survival after surgical resection. Subgroup analysis was performed according to Ki-67 and MVI to compare the recurrence free survival. Results:Multivariate Cox regression analysis showed that patients with proportion of Ki-67 positive cells ≥27.5% ( HR=2.073, 95% CI: 1.433-3.000, P<0.001) and MVI positive ( HR=2.339, 95% CI: 1.584-3.456, P<0.001) had increased risk of recurrence after radical resection. The 1-year cumulative recurrence free survival rate in the simple operation group was 70.0%, and there was no significant difference compared with 67.9% in the operation + TACE group( χ 2=0.08, P=0.774). Subgroup analysis: in the low expression of Ki-67 combined with negative MVI group ( n=128), the cumulative recurrence free survival rate of one year after operation in the simple operation group ( n=84) was 91.7%, which was significantly higher than 72.7% in the operation + TACE group ( n=44)( χ 2=8.22, P=0.004). There was no significant difference in the 1-year cumulative recurrence free survival rate between the simple operation group and the operation + TACE group (both P>0.05) in patients of Ki-67 high expression combined with MVI negative or Ki-67 low expression combined with MVI positive. In the Ki-67 high expression combined with MVI positive group ( n=107), the cumulative one-year recurrence free survival rate in the simple operation group ( n=62) was 40.3%, which was significantly lower than 60.0% in the operation + TACE group ( n=45)(χ 2=4.22, P=0.040). Conclusion:High expression of Ki-67 (≥27.5%) combined with positive MVI are the prediction factors for postoperative TACE treatment. Low expression Ki-67 (<27.5%) combined with negative MVI was contraindicated for postoperative TACE treatment.

Background/Aims: Acute-on-chronic liver failure (ACLF) is a catastrophic illness. Few studies investigated the prognostic value of vitamin D-binding protein (VDBP) for hepatitis B virus (HBV)-related ACLF (HBV-ACLF) resulted in conflicting results. Methods: Two prospective HBV-ACLF cohorts (n=287 and n=119) were enrolled to assess and validate the prognostic performance of VDBP. Results: VDBP levels in the non-survivors were significantly lower than in the survivors (P<0.001). Multivariate Cox regression demonstrated that VDBP was an independent prognostic factor for HBV-ACLF. The VDBP level at admission gradually decreased as the number of failed organs increased (P<0.001), and it was closely related to coagulation failure. The areas under the receiver operating characteristic curve (AUCs) of the Child-Pugh-VDBP and chronic liver failuresequential organ failure assessment (CLIF–SOFA)-VDBP scores were significantly higher than those of Child-Pugh (P<0.001) and CLIF-SOFA (P=0.0013). The AUCs of model for end-stage liver disease (MELD)-VDBP were significantly higher than those of MELD (P= 0.0384) only in the case of cirrhotic HBV-ACLF patients. Similar results were validated using an external multicenter HBV-ACLF cohort. By longitudinal observation, the VDBP levels gradually increased in survivors (P=0.026) and gradually decreased in non-survivors (P<0.001). Additionally, the VDBP levels were found to be significantly decreased in the deterioration group (P=0.012) and tended to be decreased in the fluctuation group (P=0.055). In contrast, they showed a significant increase in the improvement group (P=0.036). Conclusions The VDBP was a promising prognostic biomarker for HBV-ACLF. Sequential measurement of circulating VDBP shows value for the monitoring of ACLF progression.

Objective:To study combined adjuvant transcatheter arterial chemoembolization (TACE) with anti-tumor drug treatment on early hepatocellular carcinoma (HCC) recurrence in patients with microvascular invasion (MVI) after partial hepatectomy with curative intent.Methods:The clinical and pathological data of 169 patients with HCC who underwent partial hepatectomy with curative intent from January 2015 to December 2018 at the Affiliated Hospital of Qingdao University were retrospectively analyzed. MVI was diagnosed by postoperative histopathology. There were 147 males and 22 females, with the median age 56 years(ranged 32-79 years). The patients were divided into surgery group ( n=62, patients who did not receive adjuvant therapy), TACE group ( n=42, patients who only received TACE) and combined group ( n=65, patients who received TACE with anti-tumor drug) according to the therapies after resection. Patients in each group were further divided into grade M1 (mild) and grade M2 (severe) subgroups according to the severity of MVI. All patients were followed-up for observing tumor recurrence. The relapse-free survival in the three groups were compared using the Kaplan-Meier method and the log-rank test was used to compare the tumor-free survival rates. Results:The tumor-free survival rates of 169 patients at 1 and 2 years after operation were 59.2% and 40.8%. The tumor-free survival rates at 1 and 2 years after operation were 45.2% and 25.8% in surgery group, 61.9% and 40.5% in TACE group, 70.8% and 52.3% in combined group respectively. The differences among the three groups were significant: TACE group was better than surgery group, and combined group was better than TACE group, combined group was better than surgery group (all P<0.05). In TACE group and combined group, tumor-free survival rates of M1patients better than M2 patients, and the difference was significant ( P<0.05). Among M1 patients and M2 patients, tumor-free survival rates of combined group patients were better than surgery group and TACE group, the difference was significant (all P<0.05). The cumulative tumor-free survival rate was not significantly affected by different antineoplastic agents. Conclusion:Adjuvant TACE reduced the early recurrence rate of HCC patients with MVI. Adjuvant TACE combined with anti-tumor drug further reduced early tumor recurrence.