With the continuous development of liver transplantation technology, liver transplantation has been proven to be an effective treatment for end-stage liver disease, and the number of liver transplantation performed in China has been increasing year by year. Currently, the main sources of donor livers are from donation after brain death and donation after cardiac death. Although these have expanded the sources compared to the early days of liver transplantation, China, being a country with a high prevalence of liver diseases, still faces a significant gap between the demand and supply of organs, which has become the most important factor restricting the conduct of liver transplantation. Many donor livers, due to underlying diseases, prolonged ischemia, and other "marginal" factors, may lead to graft dysfunction and other complications after transplantation when using expanded criteria marginal donor livers, which can severely affect the recipient's prognosis and may even result in the discard of marginal donor livers. In order to increase the utilization rate of grafts, reduce the incidence of severe complications after liver transplantation, and improve the long-term prognosis of recipients, preoperative quality assessment of donor livers, selection of appropriate perfusion and preservation methods, and monitoring of relevant indicators in the perfusate should be performed. The above measures can, to a certain extent, expand the donor pool, predict and reduce postoperative complications through intervention, extend the overall survival time of patients, enhance the overall effectiveness of liver transplantation, and reduce the waiting time for end-stage patients to receive a transplant.

Background::Heterotaxy (HTX) is a thoracoabdominal organ anomaly syndrome and commonly accompanied by congenital heart disease (CHD). The aim of this study was to analyze rare copy number variations (CNVs) in a HTX/CHD cohort and to examine the potential mechanisms contributing to HTX/CHD.Methods::Chromosome microarray analysis was used to identify rare CNVs in a cohort of 120 unrelated HTX/CHD patients, and available samples from parents were used to confirm the inheritance pattern. Potential candidate genes in CNVs region were prioritized via the DECIPHER database, and PNPLA4 was identified as the leading candidate gene. To validate, we generated PNPLA4-overexpressing human induced pluripotent stem cell lines as well as pnpla4-overexpressing zebrafish model, followed by a series of transcriptomic, biochemical and cellular analyses. Results::Seventeen rare CNVs were identified in 15 of the 120 HTX/CHD patients (12.5%). Xp22.31 duplication was one of the inherited CNVs identified in this HTX/CHD cohort, and PNPLA4 in the Xp22.31 was a candidate gene associated with HTX/CHD. PNPLA4 is expressed in the lateral plate mesoderm, which is known to be critical for left/right embryonic patterning as well as cardiomyocyte differentiation, and in the neural crest cell lineage. Through a series of in vivo and in vitro analyses at the molecular and cellular levels, we revealed that the biological function of PNPLA4 is importantly involved in the primary cilia formation and function via its regulation of energy metabolism and mitochondria-mediated ATP production. Conclusions::Our findings demonstrated a significant association between CNVs and HTX/CHD. Our data strongly suggested that an increased genetic dose of PNPLA4 due to Xp22.31 duplication is a disease-causing risk factor for HTX/CHD.

Objective We tried to investigate the effects of human umbilical cord mesenchymal stem cell exosomes(hUCMSC-Exos)on the proliferation of human dermal papilla cells(HDPCs)and the mechanism of hUCMSC-Exos promoting hair growth.Methods HDPCs were isolated using two-step enzymatic method and cul-tured in vitro.Human umbilical cord mesenchymal stem cells(hUC-MSCs)were cultured.Cell culture supernatant was collected,and exosomes were isolated and extracted using high-speed centrifugation.Electron microscopy,particle size,and surface marker identification were performed on them.Dihydrotestosterone(DHT)induces HDPCs and establishment of an androgenic alopecia cell model.Co-culture hUCMSC-Exos with HDPCs,cell proliferation experiment(EdU)was used to detect the relative activity of induced HDPCs.Real-time qPCR was used to detect the expression level of alkaline phosphatase(ALP),and Western blot was used to detect β-catenin,Wnt10b,GSK-3β expression at the protein level.Results The obtained primary HDPCs,hUC-MSCs,and hUCMSC-Exos were all conformed to the characteristics of dermal papilla cells,mesenchymal stem cells,and exosomes.The num-ber of EdU positive cells significantly increased,and exosomes could effectively promote the proliferation of HDPCs(P<0.05),enhance the vitality of HDPCs and alleviate the damage caused by DHT(P<0.05).Real-time qPCR showed that exosomes could enhance the expression level of ALP gene(P<0.05)and hair follicle induction ability.Western Blot confirmation β-catenin,Wnt10b,GSK-3β were differences in expression at the protein level(P<0.05).Conclusions HUCMSC-Exos could promote DHT induced proliferation of HDPCs,enhance their hair follicle regeneration and repair ability,and its mechanism may be related to the activation of Wnt/β-catenin signaling pathway.

Objective:To investigate the risk factors of intraspinal cement leakage in the treatment of osteoporotic vertebral compression fracture by percutaneous vertebroplasty (PVP).Methods:From January 2018 to December 2021, 156 patients with OVCF who received surgical treatment in Beijing Tongzhou Integrated Traditional Chinese and Western Medicine Hospital were enrolled in this retrospective case-control study. The postoperative CT imaging results were analyzed, and the patients were divided into two groups according to the occurrence of intraspinal cement leakage: leakage group ( n=28) and non-leakage group ( n=128). Measurement data was expressed as mean±standard deviation ( ± s), and t-test was used for comparison of visual analogue score (VAS) between groups; the count data was expressed as the number of cases and percentage (%); univariate and multivariate Logistic regression analysis were used to evaluate the risk factors of postoperative intraspinal cement leakage. Results:All the patients were treated by PVP successfully, without obvious adverse reactions and serious complications occurred during and after the operation. Univariate Logistic regression analysis showed that higher bone mineral density ( P=0.005), OVCF combined with posterior vertebral wall injury ( P<0.001) and higher bone cement dosage ( P=0.013) were the risk factors leading to intraspinal cement leakage. Multivariate Logistic regression analysis showed that higher bone mineral density ( P=0.009, 95% CI: 0.152-0.762, OR=0.340), OVCF combined with posterior vertebral wall injury ( P=0.001, 95% CI: 2.134-15.780, OR=5.803), and higher bone cement dosage ( P=0.005, 95% CI: 1.175-2.505, OR=1.715) were the independent risk factors of intraspinal cement leakage. Conclusion:Intraspinal cement leakage was common complication after PVP. Higher bone mineral density, OVCF combined with posterior vertebral wall injury, and higher bone cement dosage were the independent risk factors affecting intraspinal cement leakage.

Objective:To explore the changes in serum levels of miR-499 and miR-362 in patients with advanced non-small cell lung cancer (NSCLC) and their relationship with prognosis.Methods:A total of 103 patients with advanced NSCLC at Shanghai University of Medicine & Health Sciences Affiliated Chongming Hospital from January 2020 to October 2021 were selected as the NSCLC group, and 100 healthy volunteers who underwent physical examinations at our hospital during the same period were selected as the control group. Fluorescent quantitative PCR was used to determine and compare the levels of serum miR-499 and miR-362 in the two groups, and the relationship between the two indexes and different clinical characteristics of NSCLC patients was analyzed. According to the clinical outcome of 2-year follow-up, the patients were divided into survival group and death group, and the levels of serum miR-499 and miR-362 were compared between the two groups. The predictive value of miR-499 and miR-362 levels on the prognosis of advanced NSCLC patients were analyzed using receiver operator characteristic (ROC) curves.Results:The serum miR-499 level in the NSCLC group (0.34±0.10) was lower than that in the control group (1.25±0.21), while the miR-362 level (1.13±0.27) was higher than that in the control group (0.63±0.15) ( t=18.26, P<0.001; t=16.32, P<0.001). There were statistically significant differences in serum miR-499 and miR-362 levels among patients with different degrees of differentiation ( t=11.12, P<0.001; t=16.35, P<0.001), TNM staging ( t=13.64, P=0.002; t=8.73, P=0.010) and lymph node metastasis ( t=10.02, P=0.003; t=9.65, P=0.004). The serum miR-499 level in the death group ( n=77) (0.24±0.06) was lower than that in the survival group ( n=26) (0.35±0.09), while the miR-362 level (1.54±0.32) was higher than that in the survival group (1.08±0.21), with statistically significant differences ( t=8.06, P=0.006; t=8.67, P=0.005). ROC curve analysis showed that the sensitivity of miR-499 and miR-362 in predicting the prognosis of advanced NSCLC patients was 73.46% and 75.85%, respectively, with specificity of 64.42% and 65.61%, AUC of 0.739 (95% CI: 0.662-0.805) and 0.743 (95% CI: 0.640-0.793) ; the sensitivity, specificity, and AUC of serum miR-499 combined with miR-362 in predicting the prognosis of advanced NSCLC patients were 87.63%, 85.34%, and 0.875 (95% CI: 0.698-0.897), respectively; the combined prediction of miR-499 and miR-362 for AUC area was higher than the individual prediction ( Z=4.83, P=0.013; Z=5.17, P=0.009) . Conclusion:Advanced NSCLC patients show significant abnormal serum level of miR-499 and miR-362, and as the severity of the disease progressed, the serum level of miR-499 is downregulated more significantly and miR-362 is upregulated more significantly. The combined detection of miR-499 and miR-362 levels has certain predictive value for the prognosis of advanced NSCLC patients.

Objective:To quantify the associations between periconceptional maternal homocysteine (HCY) and offspring′s birth weight and risk of small for gestational age (SGA) infant.Methods:The 19 984 mother-child pairs in this prospective cohort study were recruited from the Shanghai preconception cohort; the infants were delivered from 1 st September 2016 to 11 th November 2022. A standardized questionnaire was used to collect the mothers′ demographic information, medical history, dietary supplement use, and maternal complications during pregnancy, and their serum samples were collected. Serum HCY, folate, and vitamin B 12 were measured using chemiluminescent microparticle immunoassay based on serum sample drawn at enrollment. Birth weight data were obtained from medical records. Multiple imputation methods were applied to handle missing data in key variables. Multivariable linear regression and Poisson regression models were used to analyze the relationship between maternal HCY concentration during the periconceptional period and the birth weight and SGA risk of the offspring. Results:A total of 9 452 pairs were enrolled preconceptionally and the remaining 10 532 pairs were enrolled in early pregnancy. The proportion of mothers whose pregnancy age was greater than 35 years was 9.2% (1 832/19 984), the proportion of primiparous women was 76.5% (15 283/19 984), the proportion of pre-pregnancy overweight and obesity was 14.0% (2 804/19 984), the proportion of using folic acid supplements before pregnancy was 21.4% (4 272/19 984), and the proportion of those who supplemented with folic acid during early pregnancy was 85.2% (8 976/10 532); gestational diabetes mellitus was in 6.2% (1 245/19 984), gestational hypertensive syndrome in 3.6% (711/19 984). The birth weight of the offspring was (3 297±468) g, and there were 1 962 SGA children (9.8%). The HCY concentration in the overall population in appropriate for gestational age during the periconceptional period was (7.9±3.2) μmol/L, with (8.3±3.7) μmol/L in the preconception subgroup and (7.3±2.4) μmol/L in the early pregnancy subgroup. After adjustment for the covariates of perinatal demographic information, adverse pregnancy outcomes, serum folate and vitamin B 12, increased maternal periconceptional HCY was significantly associated with lower offspring birth weight ( β=-2.30, 95% CI -4.43--0.16, P=0.035). Only the early pregnancy subgroup was significantly associated with lower offspring birth weight ( β=-7.39, 95% CI-11.50--3.21, P<0.001). No association was found between peripregnancy HCY and offspring SGA risk. However, elevated HCY in early pregnancy was associated with an increased risk of SGA in the offspring ( RR=1.05, 95% CI 1.01-1.08, P=0.002). Periconceptional vitamin B 12 was a mediator of the association between HCY and offspring birth weight, accounting for 16.5%, 41.2% and 5.4% of its total effect in the overall periconceptional population, the pre-pregnancy subgroup and the early pregnancy subgroup, respectively. Conclusions:Maternal periconceptional HCY level is associated with lower birth weight in offspring, but not with the risk of SGA. Elevated maternal HCY in early pregnancy subgroup may be associated with increased risk of SGA in offspring.

Objective:To investigate the value of systemic inflammatory response syndrome (SIRS), pediatric sequential organ failure assessment (pSOFA) and pediatric critical illness score (PCIS) in predicting mortality of pediatric sepsis in pediatric intensive care units (PICU) from Southwest China.Methods:This was a prospective multicenter observational study. A total of 447 children with sepsis admitted to 12 PICU in Southwest China from April 2022 to March 2023 were enrolled. Based on the prognosis, the patients were divided into survival group and non-survival group. The physiological parameters of SIRS, pSOFA and PCIS were recorded and scored within 24 h after PICU admission. The general clinical data and some laboratory results were recorded. The area under the curve (AUC) of the receiver operating characteristic curve was used to compare the predictive value of SIRS, pSOFA and PCIS in mortality of pediatric sepsis.Results:Amongst 447 children with sepsis, 260 patients were male and 187 patients were female, aged 2.5 (0.8, 7.0) years, 405 patients were in the survival group and 42 patients were in the non-survival group. 418 patients (93.5%) met the criteria of SIRS, and 440 patients (98.4%) met the criteria of pSOFA≥2. There was no significant difference in the number of items meeting the SIRS criteria between the survival group and the non-survival group (3(2, 4) vs. 3(3, 4) points, Z=1.30, P=0.192). The pSOFA score of the non-survival group was significantly higher than that of the survival group (9(6, 12) vs. 4(3, 7) points, Z=6.56, P<0.001), and the PCIS score was significantly lower than that of the survival group (72(68, 81) vs. 82(76, 88) points, Z=5.90, P<0.001). The predictive value of pSOFA (AUC=0.82) and PCIS (AUC=0.78) for sepsis mortality was significantly higher than that of SIRS (AUC=0.56) ( Z=6.59, 4.23, both P<0.001). There was no significant difference between pSOFA and PCIS ( Z=1.35, P=0.176). Platelet count, procalcitonin, lactic acid, albumin, creatinine, total bilirubin, activated partial thromboplastin time, prothrombin time and international normalized ratio were all able to predict mortality of sepsis to a certain degree (AUC=0.64, 0.68, 0.80, 0.64, 0.68, 0.60, 0.77, 0.75, 0.76, all P<0.05). Conclusion:Compared with SIRS, both pSOFA and PCIS had better predictive value in the mortality of pediatric sepsis in PICU.

Objective:To analyze the distribution of clopidogrel metabolism-related gene variability in Kawasaki disease (KD) children with coronary artery lesions (CAL) across different age groups and the impact of genetic variability on the efficacy of clopidogrel antiplatelet therapy.Methods:A retrospective cohort study was conducted. Clinical data were collected from 46 KD children with CAL who were hospitalized in the Cardiovascular Center of Children′s Hospital of Fudan University between January 2021 and August 2022 and were treated with clopidogrel, including gender, age, body mass index, course of KD, CAL severity grade, and baseline platelet count. According to their age, the children were divided into ≥2-year-old group and <2-year-old group. Their platelet responsiveness was assessed by adenosine diphosphate-induced platelet inhibition rate (ADPi) calculated via thromboelastography, and children were categorized into high on-treatment platelet reactivity (HTPR) and normal on-treatment platelet reactivity (NTPR) groups. Genotypes of CYP2C19, PON1 and ABCB1 were detected. The t test, one-way analysis of variance and Chi-square test were used for intergroup comparison. Results:Among the 46 KD children with CAL, 34 were male and 12 were female; 37 were ≥2-year-old and 9 were <2-year-old; 25 cases were in the HTPR group and 21 cases were in the NTPR group, with 19 HTPR and 18 NTPR in the ≥2-year-old group, and 6 HTPR and 3 NTPR in the <2-year-old group. Genetic analysis showed that 92 alleles among the 46 children, with frequencies of CYP2C19*1, CYP2C19*2, CYP2C19*3, CYP2C19*17, PON1 192Q, PON1 192R, ABCB1 3435C, ABCB1 3435T at 59% (54/92), 32% (29/92), 9% (8/92), 1% (1/92), 36% (36/92), 64% (59/92), 63% (58/92) and 37% (34/92), respectively. Analysis of the impact of genotype on ADPi revealed that in children aged ≥2 years, those with CYP2C19*1/*3 genotype had significantly lower ADPi than those with CYP2C19*1/*1 genotype ((34±15)% vs. (61±29)%, t=2.18, P=0.036). There were also no significant difference in ADPi among children with PON1 192Q homozygous, PON1 192R heterozygote and PON1 192R homozygous genotypes ((40±22)% vs. (52±33)% vs. (65±27)%, F=2.17, P=0.130), or among those with ABCB1 3435C homozygous, ABCB1 3435T heterozygote and ABCB1 3435T homozygous genotypes ((55±34)% vs. (60±27)% vs. (49±24)%, F=0.33, P=0.719). In <2-year-old group, there were no significant differences in ADPi across CYP2C19*1/*1, CYP2C19*1/*2 and CYP2C19*2*2 genotypes ((40±20)% vs. (53±37)% vs. (34±16)%, F=0.37, P>0.05). There were no significant differences in ADPi across CYP2C19*1/*1 and CYP2C19*1/*3 genotypes ((44±27)% vs. (42±20)%, t=0.08, P>0.05). There were no significant differences in ADPi across PON1 192Q homozygous, PON1 192R heterozygote and PON1 192R homozygous genotypes (45% vs. (55±27)% vs. (24±5)%, F=1.83, P>0.05). There were no significant differences in ADPi across ABCB1 3435C homozygous, ABCB1 3435T heterozygote and ABCB1 3435T homozygous genotypes ((36±16)% vs. (50±35)% vs. 45%, F=0.29, P>0.05). The risk analysis of HTPR in different genotypes revealed that in children aged ≥2 years, carrying at least 1 or 2 loss-of-function alleles of CYP2C19 was a risk factor for HTPR ( OR=4.69, 10.00, 95% CI 1.11-19.83, 0.84-119.32, P=0.033, 0.046, respectively), and PON1 192R homozygosity and carrying at least one PON1 192R allele were protective factors against HTPR ( OR=0.08, 0.13, 95% CI 0.01-0.86, 0.01-1.19, P=0.019, 0.043, respectively). Conclusion:KD children aged ≥2 years carrying CYP2C19 loss-of-function alleles and PON1 192Q are more likely to develop HTPR.

Objective:To construct a social loss indicator system based on the perspective of epidemic prevention and control against background of emerging major infectious diseases at the national, hospital, and individual levels, and to provide decision-making basis for public general hospitals to formulate prevention and control strategies for emerging major infectious diseases.Methods:Literatures published before December 23, 2020 were retrieved from CNKI, Wanfang, VIP, Web of Science, Google Scholar with such keywords as " infectious diseases" " public health emergencies" " social losses" " COVID-19" " evaluation". Then a preliminary social loss indicator system of public general hospitals for emerging major infectious diseases was constructed from such dimensions as country, hospital, and individual, by implementing literature analysis and expert consultation. Based on such a preliminary system, three questionnaires were designed for the country, hospitals, and individuals and the corresponding data were collected from December 25, 2021 to May 20, 2022. In the end, a factor analysis was made on the questionnaire data to optimize the social loss indicator system and determine the weights of each indicator.Results:The social loss indicator system consists of 14 level-1 indicators and 60 level-2 indicators. Level-1 indicators at country aspect consisted of government governance capacity, direct economic losses, social security, online public opinion, indirect economic losses, and international cooperation; indicators at hospital aspect consisted of hospital manpower, material resources, and information resources, hospital services, and hospital operations; indicators at the individual aspect consisted of physical health, psychological and social health, and external environment. Level-1 indicators of the highest weight at all aspects were social security, manpower, material and information resources, as well as physiological health. And the level-2 indicators of the highest weight were high medical expenses, overwork for staff other than doctors and nurses and medical insurance.Conclusions:The social loss indicator system for emerging major infectious diseases constructed in this study proves scientific and reasonable, helpful for the formulation of prevention and control strategies of public general hospitals.

Objective:To study the effects of Zuogui Pills on the expressions of miR-133b-3p and RhoA in osteoclasts of postmenopausal osteoporosis rats; To discuss its potential mechanism.Methods:SD female rats were randomly divided into normal group, model group, sham-operation group, and Zuogui Pills group using a random number table method, with 6 rats in each group. The model group and Zuogui Pills group were treated with oophorectomy to construct a rat model of osteoporosis. Zuogui Pills group was orally administered with Zuogui Pills decoction at a concentration of 10 g/kg for 12 consecutive weeks. Colorimetric method was used to measure the serum calcium and phosphorus levels of rats, and ELISA method was used to detect ALP levels. Bone density meter was used to measure the bone density of the femurs of rats in each group. The osteoclast of each group were cultured, and the expressions of RANKL and RUNX2 protein were detected by Western blot. MiRNA sequencing and differential expression analysis were performed on bone tissues of rats. Osteoclasts were treated with miR-133b-3p mimic and its negative control. The cell proliferation activity of osteoclasts was detected by cell counting kit-8 (CCK-8). The osteoclast differentiation activity was detected by the tartrate-resistant acid phosphatase staining. The dual-luciferase reporter assay was used to detect the relationship between miR-133b-3p and RhoA. The "rescue" experiment of miR-133b-3p mimic and RhoA co-expression were used to study the molecular regulatory mechanism of Zuogui Pills on osteoclast activity.Results:Compared with the model group, the bone mineral density of Zuogui Pills group significantly increased ( P<0.05, P<0.01), the levels of calcium and phosphorus in serum increased, the level of alkaline phosphatase ALP decreased ( P<0.05), the expression of RANKL protein decreased, and the expression of RUNX2 protein increased. Sequencing results showed that rno-miR-133b-3p was down-regulated in osteoclasts of postmenopausa osteoporosis rats treated with Zuogui Pills with the maximum difference ( P<0.01). Q-PCR results showed that the expression of miR-133b-3p in osteoclasts of Zuogui Pills group was significantly lower than that of the model group. The upregulation of miR-133b-3p could significantly promote the cell proliferation and differentiation of osteoclasts. RhoA overexpression could reverse the excessive proliferation and differentiation of osteoclasts caused by miR-133b-3p overexpression. Conclusions:RhoA is the target gene regulated by miR-133b-3p. Zuogui Pills can inhibit the activity of osteoclasts by regulating miR-133b-3p/RhoA axis, relieving the symptoms of osteoporosis.