Objective To explore the construction of heart preservation model of empty beating donor based on extracorporeal membrane oxygenation (ECMO). Methods From January 2022 to August 2023, 20 Guangxi Bama miniature pigs weighing 25-30 kg were selected, half male and half female. Under general anesthesia and heparinization, a midline thoracotomy was performed. The pericardium was cut after freeing the anterior and posterior vena cavae, and a perfusion needle was inserted near the brachiocephalic artery in the ascending aorta, connected to a blood collection bag to collect 500-600 mL of blood. The anterior and posterior vena cavae were ligated, the aorta was blocked and perfused with HTK solution to stop the heart beating. The superior and inferior vena cavae were cut off, the right pulmonary vein was decompressed, the aorta and left and right pulmonary arteries and veins were cut off, and the whole heart was removed. An ECMO device was used to continuously perfuse a cardioprotective solution mainly composed of oxygenated warm blood, maintaining the isolated pig heart beating for 8 hours, monitoring (once/hour) ECMO perfusion parameters, blood gas indicators, perfusate electrolytes, inflammatory factors, myocardial enzymes, myoglobin, and troponin levels. Myocardial tissue was taken for hematoxylin-eosin (HE) staining to observe myocardial cell damage and evaluate the quality of heart preservation. Results Among the 20 isolated beating pig hearts, 17 successfully resumed beating, 3 experienced ventricular fibrillation, resuscitated after intracardiac electrical defibrillation, and all 20 pig hearts successfully beat for 8 hours. There was no statistical difference in ECMO perfusion parameters, blood gas indicators, perfusate electrolytes, and inflammatory factors at each time point (P>0.05). There were statistical increases in myocardial enzymes, myoglobin, and troponin levels (P<0.05). HE staining results suggested that there was no severe myocardial damage. Conclusion ECMO technology can be used for pig heart preservation with good results, and this study provides experimental evidence for improving heart preservation research in clinical heart transplantation.

Objective:To explore the guiding significance of Expanded Carrier Screening (ECS) for the fertility process of individuals undergoing Assisted Reproductive Technology (ART) in Dalian.Methods:A retrospective analysis was conducted on the ECS results of patients who visited the Dalian Maternal and Child Health Care Center (Group) from January 2023 to June 2024. The study included the screening of 155 monogenic diseases and the ART status of carrier couples.Results:Among the 1 384 patient samples, 490 carriers were identified, yielding a detection rate of 35.41% (490/1 384). A total of 100 diseases and 91 pathogenic genes were detected. The most frequently identified disease was autosomal recessive hearing loss type 4 (SLC26A4), with a carrier rate of 3.54% (49/1384). For the 490 carriers, their partners were recalled for testing and, 322 partners participated. Eight high-risk couples were identified, accounting for 2.48% (8/322). Among the 322 couples, 4 used artificial insemination by husband (AIH, 1.24%, 4/322), 166 used in vitro fertilization (IVF, 51.55%, 166/322), 139 used intracytoplasmic sperm injection (ICSI, 43.17%, 139/322), and 1 underwent preimplantation genetic testing for monogenic diseases (PGT-M, 0.31%, 1/322). Twelve couples (3.73%, 12/322) did not undergo ART, were not pregnant, or chose gamete donation. Among the eight high-risk couples, three underwent IVF, four underwent ICSI, and one underwent PGT-M. One couple conceived through ICSI, and the fetus was followed up. Amniotic fluid Sanger sequencing revealed that the fetus had compound heterozygous mutations in the PAH gene, with two suspected pathogenic variants at c.532G>A and c.1174T>A. One couple undergoing PGT-M currently has one retrieved oocyte and zero usable blastocysts. The embryo carries a paternal mutation and is aneuploid.Conclusions:This study not only identified the common pathogenic diseases in Dalian, providing a reference for clinical treatment, but also validated the critical significance of ECS for individuals undergoing ART.

Objective:To explore the guiding significance of Expanded Carrier Screening (ECS) for the fertility process of individuals undergoing Assisted Reproductive Technology (ART) in Dalian.Methods:A retrospective analysis was conducted on the ECS results of patients who visited the Dalian Maternal and Child Health Care Center (Group) from January 2023 to June 2024. The study included the screening of 155 monogenic diseases and the ART status of carrier couples.Results:Among the 1 384 patient samples, 490 carriers were identified, yielding a detection rate of 35.41% (490/1 384). A total of 100 diseases and 91 pathogenic genes were detected. The most frequently identified disease was autosomal recessive hearing loss type 4 (SLC26A4), with a carrier rate of 3.54% (49/1384). For the 490 carriers, their partners were recalled for testing and, 322 partners participated. Eight high-risk couples were identified, accounting for 2.48% (8/322). Among the 322 couples, 4 used artificial insemination by husband (AIH, 1.24%, 4/322), 166 used in vitro fertilization (IVF, 51.55%, 166/322), 139 used intracytoplasmic sperm injection (ICSI, 43.17%, 139/322), and 1 underwent preimplantation genetic testing for monogenic diseases (PGT-M, 0.31%, 1/322). Twelve couples (3.73%, 12/322) did not undergo ART, were not pregnant, or chose gamete donation. Among the eight high-risk couples, three underwent IVF, four underwent ICSI, and one underwent PGT-M. One couple conceived through ICSI, and the fetus was followed up. Amniotic fluid Sanger sequencing revealed that the fetus had compound heterozygous mutations in the PAH gene, with two suspected pathogenic variants at c.532G>A and c.1174T>A. One couple undergoing PGT-M currently has one retrieved oocyte and zero usable blastocysts. The embryo carries a paternal mutation and is aneuploid.Conclusions:This study not only identified the common pathogenic diseases in Dalian, providing a reference for clinical treatment, but also validated the critical significance of ECS for individuals undergoing ART.

Objective:To study the impact of the age-adjusted Charlson comorbidity index (ACCI) on the prognosis of patients with hilar cholangiocarcinoma following laparoscopic surgical resection.Methods:Clinical data of 136 patients with hilar cholangiocarcinoma undergoing laparoscopic surgery at Zhengzhou University People's Hospital between January 2013 and January 2018 were retrospectively analyzed, including 81 males and 55 females, aged (63.6±9.8) years. Patients were divided into two groups based on the median ACCI score of 4.0: the high ACCI group (ACCI>4.0, n=49) and low ACCI group (ACCI≤4.0, n=87). The prognosis was compared between the two group. Univariate and multivariate Cox regression analyses were performed to analyze the effect of ACCI on survival after laparoscopic surgery. Results:The 1- and 3-year cumulative survival rates in low ACCI group were 87.4% and 48.3%, respectively, compared to 53.1% and 4.1% in high ACCI group ( χ2=27.97, P<0.001). Univariate Cox regression analysis indicated that ACCI >4.0 was associated with prognosis ( HR=3.73, 95% CI: 2.44-5.68, P<0.001). Multivariate Cox regression analysis also indicated that ACCI >4.0 was associated with an increased risk of postoperative mortality in patients with hilar cholangiocarcinoma ( HR=2.69, 95% CI: 1.65-4.37, P<0.001). Conclusion:The ACCI is a significant risk factor for survival of patients with hilar cholangiocarcinoma following laparoscopic surgery, which could facilitate a precise preoperative assessment of patient status and choice of surgical approach.

Objective:To investigate the influencing factors for microvascular invasion (MVI) in hepatocellular carcinoma based on three-dimensional visualization and the construction of its nomogram model.Methods:The retrospective cohort study method was conducted. The clinico-pathological data of 190 patients with hepatocellular carcinoma who were admitted to Henan University People′s Hospital from May 2018 to May 2021 were collected. There were 148 males and 42 females, aged (58±12)years. The 190 patients were randomly divided into the training set of 133 cases and the validation set of 57 cases by the method of random number table in the ratio of 7:3. The abdominal three-dimensional visualization system was used to characterize the tumor morphology and other imaging features. Observation indicators: (1) analysis of influencing factors for MVI in hepatocellular carcinoma; (2) construction and evaluation of nomogram model of MVI in hepatocellular carcinoma. Measurement data with normal distribution were expressed as Mean± SD, and independent sample t test was used for comparison between groups. Measurement data with skewed distribution were expressed as M( Q1, Q3), and non-parametric rank sum test was used for comparison between groups. Count data were expressed as absolute numbers, and the chi-square test was used for comparison between groups. Corresponding statistical methods were used for univariate analysis. Binary Logistic regression model was used for multivariate analysis. Receiver operator characteristic (ROC) curves were plotted, and the nomogram model was assessed by area under the curve (AUC), calibration curve, and decision curve. Results:(1) Analysis of influencing factors for MVI in hepatocellular carcinoma. Among 190 patients with hepatocellular carcinoma, there were 97 cases of positive MVI (including 63 cases in the training set and 34 cases in the validation set) and 93 cases of negative MVI (including 70 cases in the training set and 23 cases in the validation set). Results of multivariate analysis showed that alpha-fetoprotein, vascular endothelial growth factor, tumor volume, the number of tumors, and tumor morphology were independent factors affecting the MVI of patients with hepatocellular carcinoma ( odds ratio=5.06, 3.62, 1.00, 2.02, 2.59, 95% confidence interval as 1.61-15.90, 1.28-10.20, 1.00-1.01, 1.02-3.98, 1.03-6.52, P<0.05). (2) Construction and evaluation of nomogram model of MVI in hepatocellular carcinoma. The results of multivariate analysis were incorporated to construct a nomogram prediction model for MVI of hepatocellular carcinoma. ROC curves showed that the AUC of the training set of nomogram model was 0.85 (95% confidence interval as 0.79-0.92), the optimal fractional cutoff based on the Jordon′s index was 0.51, the sensitivity was 0.71, and the specificity was 0.84. The above indicators of validation set were 0.92 (95% confidence interval as 0.85-0.99), 0.50, 0.90, and 0.82, respectively. The higher total score of the training set suggested a higher risk of MVI in hepatocellular carcinoma. The calibration curves of both training and validation sets of nomogram model fitted well with the standard curves and have a high degree of calibration. The decision curve showed a high net gain of nomogram model. Conclusions:Alpha-fetoprotein, vascular endothelial growth factor, tumor volume, the number of tumors, and tumor morphology are independent influencing factors for MVI in patients with hepatocellular carcinoma. A nomogram model constructed based on three-dimensional visualized imaging features can predict MVI in hepatocellular carcinoma.

Objective A precise microinvasive robot system coupled with a cooperative robot matrix and an end effector of a wire-controlled microvariable path robot was proposed.A puncture needle structure,including the rigid body of the outer needle,non-uniform flexible body of the inner needle,force control wire,internal imaging of the image fiber,and other components,was designed to verify the feasibility of this system.Methods By constructing the puncture structure of the puncture robot,the structural optimization design of the key components affecting the variable-path precision puncture needle was analyzed by constructing the puncture structure of a puncture robot.Based on the orthogonal experimental design method,a three-factor and three-level experiment that primarily affected the accuracy of the puncture needle was designed,namely,the starting distance of the hole center from the edge,the diameter of the hole,and the distance between the two holes.The experiment was verified and simulated using a real object.Results The displacement of the titanium-nickel needle tip had a significant relationship with the starting distance of the hole center from the edge,and the main and secondary influencing factors were as follows:starting distance of the hole center from the edge>hole diameter>hole distance.When the starting distance of the hole center from the edge was 1 mm,the diameter of the hole was 0.2 mm,and the distance between the two holes was 2.6 mm,the displacement of the titanium-nickel needle tip was the maximum value.Conclusions The experiment verified the functional applicability of the designed system and the linear elastic hysteresis characteristics of variable path puncture,providing a reference for further in vivo experiments and system optimization.

Objective To investigate the regulatory effect of Xuebijing injection on inflammatory reaction during the preservation of isolated empty beating pig hearts with extracorporeal membrane oxygenation.Methods Twelve healthy Guangxi Bama miniature pigs were randomly divided into the Xuebijing group(n=6)and normal saline group(n=6).After the models were established in the Xuebijing group,Xuebijing injection was given at a dose of 5 mL/h through micropump in membrane oxygenator.In the normal saline group,an equivalent amount of 0.9％sodium chloride injection was pumped.Continuous pumping was performed for 8 h in both groups.The time of cardiac resuscitation and perfusion pressure,heart rate,perfusion flow rate after 8 h preservation were recorded in two groups.Pathological and ultrastructural changes of myocardial tissues in the left ventricular wall of hearts with cardiac arrest were observed after 8 h preservation.Serum levels of myocardial injury markers and inflammatory cytokines were detected in two groups at the beginning(T0),2 h(T2),4 h(T4),6 h(T6)and 8 h(T8)after model establishment,respectively.The expression levels of NOD-like receptor protein 3(NLRP3),cysteinyl aspartate specific proteinase-1(Caspase-1),apoptosis-associated speck-like protein containing a CARD(ASC)messenger RNA(mRNA)in myocardial tissues were measured at T0,T2,T4,T6 and T8,respectively.Results There were no significant differences in the time of cardiac resuscitation and perfusion pressure,heart rate,perfusion flow rate after 8 h preservation between two groups(all P＞0.05).Compared with the normal saline group,the levels of lactate dehydrogenase(LDH)at T4,creatine kinase(CK),LDH and α-hydroxybutyrate dehydrogenase(α-HBDH)at T6 and T8,tumor necrosis factor(TNF)-α at T4,T6 and T8,and interleukin(IL)-6,IL-18 and IL-1 β at T0,T2,T4,T6 and T8 were lower,and the mRNA relative expression levels of NLRP3 and Caspase-1 at T2,T4 and T6,and Caspase-1 and ASC at T8 were lower in the Xuebijing group,respectively(all P＜0.05).Hematoxylin-eosin staining and transmission electron microscopy showed that the degree of myocardial injury in the Xuebijing group was slighter than that in the normal saline group.Conclusions Xuebijing injection may effectively mitigate inflammatory response and exert certain myocardial protection effect during the ECMO preservation of isolated empty beating pig hearts.

Objective:To analyze the influencing factors of abnormal 15-minute retention rate of indocyanine green (ICG R15) (≥10%) in patients with hepatocellular carcinoma, and to construct a nomogram model, and to evaluate the prediction efficiency of the nomogram model.Methods:The clinical data of 190 patients with hepatocellular carcinoma in Zhengzhou University People's Hospital from December 2017 to June 2022 were retrospectively analyzed, including 148 males and 42 females, aged (57.8±9.9) years. According to ICG R15, the patients were divided into ICG R15 normal group ( n=134, ICG R15<10%) and ICG R15 abnormal group ( n=56, ICG R15≥10%). Univariate and multivariate logistic regression were used to analyze the influencing factors of abnormal ICG R15, and the nomogram model was established. The predictive ability of the model was evaluated by receiver operating characteristic (ROC) curve and C-index, and the model was verified by calibration curve and decision analysis curve. Results:Abnormal ICG R15 group the proportion of liver cirrhosis, albumin ≤35 g/L, hemoglobin ≤110 g/L, platelet count ≤100×10 9/L, prothrombin time >13 s, alanine aminotransferase >40 U/L, aspartate aminotransferase >40 U/L, total bilirubin >34.2 μmol/L, and the largest tumor diameter >5.0 cm, spleen volume >383.1 cm 3, spleen volume to of non-tumor liver volume (SNLR) >0.276 and liver tumor volume >117.2 cm 3 were higher than that of ICG R15 normal group, and the differences were statistically significant (all P<0.05). Logistic regression analysis showed that liver cirrhosis ( OR=3.89, 95% CI: 1.28-11.80, P=0.016), spleen volume >383.1 cm 3( OR=5.17, 95% CI: 1.38-19.38, P=0.015), SNLR >0.276 ( OR=5.54, 95% CI: 1.44-21.26, P=0.013) and total bilirubin >34.2 μmol/L( OR=10.20, 95% CI: 1.88-55.39, P=0.007) increased the risk of abnormal ICG R15. A nomogram model was constructed based on the above risk factors. The C-index of the model was 0.915 (95% CI: 0.872-0.957), and the area under the ROC curve predicted by the nomogram model was 0.915 (95% CI: 0.871-0.958). The calibration curve showed that the correlation index of the abnormal ICG R15 predicted by the nomogram was similar to actual situation. Decision analysis curve showed high returns. Conclusion:Liver cirrhosis, spleen volume >383.1 cm 3, SNLR>0.276 and total bilirubin >34.2 μmol/L were indepentlent risk factors for abnormal ICG R15 in patients with hepatocellur carcinoma. The clinical prediction model of ICG R15 abnormality constructed by nomogram has good prediction efficiency, which can provide a reference for evaluating preoperative liver reserve function of patients with hepatocellular carcinoma.

Objective:To explore the application of microfluidic chip in detection of hereditary deafness-associated hotspot mutations.Methods:The dedicatedly designed and fabricated microfluidic chip was integrated with kompetitive allele-specific polymerase chain reaction amplification system, scanned via laser-excited confocal fluorescence scanner, and finally analyzed programmatically to acquire the typing results of the 23 mutation sites of the four common genes associated with hereditary hearing loss. Dried blood spots were collected from 276 neonates containing the 131 cases with hearing loss who were born in 2019 in Chengdu (deafness group) and the 145 cases with normal hearing who were born in 2020 in Chengdu (control group), and analyzed by the microfluidic chip to evaluate its clinical performance.Results:By cluster analysis, the microfluidic chip correctly analyzed the 23 positive reference samples and acquired the same typing results as their actual results, with a limit of detection of 1 mg/L. For the 276 newborn blood spots, the detection results of the microfluidic chips were confirmed to be correct by the contrasting methods. Among Deafness Group, 66 (50.4%) tested positive for the selected 23 mutation hotspots; among Control Group, 40(27.6%) were positive. Among these mutations, c.109G>A of the GJB2 gene was the most prevalent one, whose carrier rate in deafness group and control group were 46.6%(61/131) and 23.4% (34/145), respectively.Conclusions:The micro-fluidic chip system was succeeded in fulfilling the hereditary deafness-related mutation detection, and offered many advantages including high specificity, avoiding the amplicon carryover contamination, simplifying the entire experimental operation process and short detection time, so as to better meet the detection requirement of genetic testing for deafness in newborn screening and other fields.

Objective To study the iodine nutritional status of pregnant women in Foshan City,Guangdong Province,and to provide basis for iodine supplementation scientifically for them.Methods In 2017,five districts Chancheng,Nanhai,Shunde,Sanshui and Gaoming in Foshan were selected.Each monitoring site was divided into five sections according to stratified random sampling in five directions:east,west,south,north and middle.One township (street) was taken in each section,and 20 pregnant women were taken from each township (street) to detected their family salt iodine and urinary iodine levels.Results A total of 500 edible salt samples were collected from the families of pregnant women,the median of salt iodine was 24.8 mg/kg,the qualified iodized salt consumption rate was 95.8％ (479/500),and the iodized salt coverage rate was 99.0％ (495/500).There was a statistically significant difference in the iodine content of edible salt of pregnant women in diffierent districts (H =26.9,P ＜ 0.05).A total of 500 urine samples were collected from pregnant women,the median of urinary iodine was 138.9 μg/L.There was a statistically significant difference in the levels of urinary iodine content of pregnant women in different districts (H =14.5,P ＜ 0.05).There was no statistically significant difference in urinary iodine of pregnant women during different stages of pregnancy (H =2.6,P ＞ 0.05).Conclusions The iodine intake for pregnant women is in a state of deficiency (＜ 150 μg/L) in Foshan of Guangdong Province.It is necessary to further strengthen the health education of key populations,and improve the iodine nutritional status of pregnant women.