AIM: To investigate the potential of human induced pluripotent stem cells(hiPSCs)differentiating into corneal epithelial cells in the simulated limbal stem cells(LSCs)microenvironment.METHODS: The hiPSC cell lines were established in vitro, and hiPSCs were co-cultured with corneal stromal cells in transwell system, which simulated the LSC microenvironment. Bone morphogenetic protein 4(BMP4)and a specific transforming growth factor β inhibitor(SB431542)were added to improve the differentiation efficacy. The expression of corneal epithelial cell-specific markers CK3 and CK12, corneal epithelial cell precursor CK15, and the limbal stem cell markers ABCG5 were determined by immunofluorescence staining and flow cytometry.RESULTS: The hiPSCs were actively proliferated in vitro, and immunofluorescence staining showed positive stem cell-specific markers OCT4, SOX2, TRA-1-60 and NANOG. Furthermore, hiPSCs co-cultured with corneal stromal cells exhibited LSCs markers ABCG5 and corneal epithelial cell precursor markers CK15 were positive; however, corneal epithelial cell markers CK3 and CK12 were negative. With the addition of BMP4 and SB431542, hiPSCs showed positive expression of CK3, and the CK3 expression increased over the time.CONCLUSION: With the addition of SB431542 and BMP4, hiPSCs cultured in simulated LSCs microenvironment could differentiate into corneal epithelial cells.

Following the principles of evidence-based medicine,in accordance with the structure and drafting rules of standardized documents,based on literature research,according to the characteristics of chronic cough in children and issues that need to form a consensus,the TCM Guidelines for Diagnosis and Treatment of Chronic Cough in Children was formulated based on the Delphi method,expert discussion meetings,and public solicitation of opinions.The guideline includes scope of application,terms and definitions,eti-ology and diagnosis,auxiliary examination,treatment,prevention and care.The aim is to clarify the optimal treatment plan of Chinese medicine in the diagnosis and treatment of this disease,and to provide guidance for improving the clinical diagnosis and treatment of chronic cough in children with Chinese medicine.

Objectives:To investigate the impact of resting heart rate on the risk of all-cause mortality in ultra-high risk atherosclerotic cardiovascular disease(ASCVD)patients. Methods:A total of 3 645 patients with ultra-high risk ASCVD(as defined in the 2023 Chinese Lipid Management Guidelines)were screened from the 2006 to 2020 Kailuan Study cohort,and after excluding 602 patients with missing resting heart rate,3 043 patients were included in the final analysis.Patients were divided into＜68 beats/min group(n=744),68-74 beats/min group(n=786),75-80 beats/min group(n=760),and≥81 beats/min group(n=753)according to the resting heart rate.Cox proportional regression model was used to estimate the hazard ratios(HRs)and 95%CI for all-cause mortality associated with the different resting heart rate groups and every 10 beats/min increase of resting heart rate.The dose-effect relationship of resting heart rate level and all-cause mortality was assessed by a restricted cubic spline regression model.The Kaplan-Meier method was applied to calculate the cumulative all-cause mortality in different groups,and the differences were compared using log-rank test. Results:The median follow-up time was 5.81(3.46,9.64)years,there were 772(25.37%)all-cause deaths during follow up.After adjusting major confounding factors,the results showed that compared with＜68 beats/min group,the risk of all-cause mortality in 75-80 beats/min group and≥81 beats/min group increased by 24%(HR=1.24,95%CI:1.01-1.52,P=0.047)and 47%(HR=1.47,95%CI:1.20-1.81,P＜0.001),respectively;the risk of all-cause mortality in 68-74 beats/min group was similar(HR=1.06,95%CI:0.86-1.31,P=0.625).In addition,an increase of 10 beats/min in resting heart rate was associated with a 13%increase in the risk of all-cause mortality(HR=1.13,95%CI:1.07-1.19,P＜0.001).In stratified analyses,it was found that for every 10 beats/min increase in resting heart rate,women faced a higher risk of all-cause mortality than men,and patients＜65 years old faced a higher risk of all-cause mortality than patients≥65 years old.The restricted cubic spline analysis also showed that resting heart rate was linearly associated with the risk of all-cause mortality(Poverall＜0.001,Pnon-linear=0.933),and the risk increased significantly with resting heart rate＞70 beats/min. Conclusions:Increased resting heart rate is linearly associated with increased risk of all-cause mortality in patients with ultra-high risk ASCVD.The appropriate intervention cut-off point of resting heart rate for ultra-high risk ASCVD patients may be＞75 beats/min.

Objective To investigate the protective effects of HTD4010 against lipopolysaccharide(LPS)-induced septic cardiomyopathy(SCM)in mice and explore the mechanisms mediating its effect.Methods Forty-five male ICR mice were randomized equally into control group,LPS(10 mg/kg)group,and LPS+HTD4010 group(in which 2.5 mg/kg HTD4010 was injected subcutaneously at 1 h and 6 h after LPS injection).Cardiac function of the mice was evaluated by ultrasound,and pathological changes in the myocardial tissues were observed with HE staining.The levels of IL-6 and TNF-α in serum and myocardial tissues were detected using ELISA,and apoptosis of the cardiomyocytes was detected with TUNEL staining.The expression levels of the key proteins associated with apoptosis,autophagy and the AMPK/mTOR pathway in the myocardial tissues were detected using Western blotting.The ultrastructural changes of cardiac myocardial mitochondria was observed with transmission electron microscopy.Results LPS exposure caused severe myocardial damage in mice,characterized by myocardial fiber rupture,structural disorder,inflammatory cell infiltration,and mitochondrial damage.The LPS-treated mice exhibited significantly decreased cardiac LVEF and FS values,elevated IL-6 and TNF-αlevels in serum and myocardial tissue,and an increased myocardial cell apoptosis rate with enhanced expressions of Bax,p-62 and p-mTOR and lowered expressions of Bcl-2,LC3 Ⅱ/I,Beclin-1 and p-AMPK(P<0.05 or 0.01).Treatment of the septic mice with HTD4010 significantly alleviated myocardial damage,increased LVEF and FS values,reduced IL-6 and TNF-α levels in serum and myocardial tissue,decreased cardiomyocyte apoptosis,lowered myocardial expressions of Bax,p-62 and p-mTOR,and increased Bcl-2,LC3 Ⅱ/I,Beclin-1 and p-AMPK expressions(P<0.05 or 0.01).Conclusion HTD4010 can attenuate myocardial injury in SCM mice possibly by promoting autophagy via modulating the AMPK/mTOR signaling pathway.

Objectives:To investigate the association of trajectories of atherogenic index of plasma(AIP)with the risk of atherosclerotic cardiovascular disease(ASCVD). Methods:A total of 51 831 employees and retirees who participated in Kailuan Group health examination for three consecutive times from 2006 to 2010 were included in this study.AIP was calculated using the log(triglycerides[TG]/high-density lipoprotein-cholesterol[HDL-C])formula.AIP trajectory models were fitted by the SAS Proc Traj program,and according to AIP trajectory,the subjects were divided into low stability group(n=11 114),low to moderate stability group(n=21 647),medium to high stability group(n=13 659),and high stability group(n=5 411).Kaplan-Meier method was used to calculate the cumulative incidence of ASCVD in different groups and compared by log-rank test.Cox proportional risk regression model was used to analyze the effects of different AIP trajectories on ASCVD risk. Results:Finally,51 831 patients were included in the analysis.During a mean follow-up of(10.19±2.22)years,5 142(9.92%)subjects developed ASCVD,4 013(7.74%)subjects died.Cox regression analysis after adjusting for confounding factors showed:compared with the low stability group,the risk of ASCVD increased by 13%(HR=1.13,95%CI:1.04-1.23,P=0.003)and 20%(HR=1.20,95%CI:1.10-1.31,P＜0.001)and 41%(HR=1.41,95%CI:1.27-1.57,P＜0.001)in the low to moderate stability group,moderate to high stability group and high stability group,respectively,and the risk increased gradually(Ptrend＜0.001).Stratified analysis showed that the risk of ASCVD in people aged＜65 years and low-density lipoprotein cholesterol(LDL-C)＜3.4 mmol/L with long-term high levels of AIP was higher than that in people aged≥65 years and LDL-C≥3.4 mmol/L(both Pinteraction＜0.01). Conclusions:In Kailuan Study cohort,those with long-term high levels of AIP had a higher risk of ASCVD,and the risk gradually increased.In addition,we found that the risk of ASCVD in people with long-term high levels of AIP was higher in＜65 years old than in≥65 years old,and the risk of ASCVD in people with LDL-C＜3.4 mmol/L was higher than that in people with LDL-C≥3.4 mmol/L.

Objective To investigate the protective effects of HTD4010 against lipopolysaccharide(LPS)-induced septic cardiomyopathy(SCM)in mice and explore the mechanisms mediating its effect.Methods Forty-five male ICR mice were randomized equally into control group,LPS(10 mg/kg)group,and LPS+HTD4010 group(in which 2.5 mg/kg HTD4010 was injected subcutaneously at 1 h and 6 h after LPS injection).Cardiac function of the mice was evaluated by ultrasound,and pathological changes in the myocardial tissues were observed with HE staining.The levels of IL-6 and TNF-α in serum and myocardial tissues were detected using ELISA,and apoptosis of the cardiomyocytes was detected with TUNEL staining.The expression levels of the key proteins associated with apoptosis,autophagy and the AMPK/mTOR pathway in the myocardial tissues were detected using Western blotting.The ultrastructural changes of cardiac myocardial mitochondria was observed with transmission electron microscopy.Results LPS exposure caused severe myocardial damage in mice,characterized by myocardial fiber rupture,structural disorder,inflammatory cell infiltration,and mitochondrial damage.The LPS-treated mice exhibited significantly decreased cardiac LVEF and FS values,elevated IL-6 and TNF-αlevels in serum and myocardial tissue,and an increased myocardial cell apoptosis rate with enhanced expressions of Bax,p-62 and p-mTOR and lowered expressions of Bcl-2,LC3 Ⅱ/I,Beclin-1 and p-AMPK(P<0.05 or 0.01).Treatment of the septic mice with HTD4010 significantly alleviated myocardial damage,increased LVEF and FS values,reduced IL-6 and TNF-α levels in serum and myocardial tissue,decreased cardiomyocyte apoptosis,lowered myocardial expressions of Bax,p-62 and p-mTOR,and increased Bcl-2,LC3 Ⅱ/I,Beclin-1 and p-AMPK expressions(P<0.05 or 0.01).Conclusion HTD4010 can attenuate myocardial injury in SCM mice possibly by promoting autophagy via modulating the AMPK/mTOR signaling pathway.

Critical ultrasonography is widely used in ICU and has become an indispensable tool for clinicians. However, besides operator-dependency of critical ultrasonography, lack of standardized training mainly result in the physicians′ heterogenous ultrasonic skill. Therefore, standardized training as well as strict quality control plays the key role in the development of critical ultrasonography. We present this quality control standards to promote better development of critical ultrasonography.

Genomic disorders caused by pathogenic copy number variation (pCNV) have proven to underlie a significant proportion of birth defects. With technological advance, improvement of bioinformatics analysis procedure, and accumulation of clinical data, non-invasive prenatal screening of pCNV (NIPS-pCNV) by high-throughput sequencing of maternal plasma cell-free DNA has been put to use in clinical settings. Specialized standards for clinical application of NIPS-pCNV are required. Based on the discussion, 10 pCNV-associated diseases with well-defined conditions and 5 common chromosomal aneuploidy syndromes are recommended as the target of screening in this consensus. Meanwhile, a standardized procedure for NIPS-pCNV is also provided, which may facilitate propagation of this technique in clinical settings.
Aneuploidy ; Cell-Free Nucleic Acids/genetics* ; Consensus ; DNA Copy Number Variations ; Female ; High-Throughput Nucleotide Sequencing ; Humans ; Pregnancy ; Prenatal Diagnosis

Purpose: This study aimed to describe the clinical response to five-step systematic therapy (FSST) in the management of plugged ducts and mastitis. FSST was a comprehensive milk stasis dredging treatment, which contained five steps to make the milk out of the plugged duct. Methods: This retrospective study included 922 breastfeeding women, 714 with plugged ducts, and 208 with mastitis who received FSST from June to September 2017. The breast pain score, swelling degree, and range of breast induration were recorded pre-FSST and post-FSST. Results: After a single FSST, pain score and swelling degree were significantly improved (both p < .001) in all cases. After FSST, the mean breast pain relief score was 1.69 ± 0.70, whereas the mean swelling fade away degree was 1.61 ± 0.62. In the subgroup analysis, pain score and swelling degree were significantly improved (both p < .001) in the plugged ducts group and the mastitis group. The score of pain relief in the plugged ducts group was less than that in the mastitis group (1.63 ± 0.68 vs. 1.91 ± 0.70, t = 5.30; p < .001), whereas improvement of swelling fade away was greater in the plugged ducts group than the mastitis group (1.65 ± 0.64 vs. 1.48 ± 0.56, t = 3.49; p = .001). The composition ratio of changes in induration range between the two groups was statistically different (Pearson χ2 = 137.87, p < .001), of which more obvious improvement in the plugged ducts group than the mastitis group (χ2 = 25.65, p < .001). Conclusion FSST can relieve pain, reduce breast swelling and range of induration, and for plugged ducts or mastitis varied degree differently.

Objective:To investigate the clinical features and prognosis of acute necrotizing encephalopathy (ANE) in children.Methods:The clinical data and follow-up information of 41 pediatric patients with ANE treated in Wuhan Children′s Hospital, Tongji Medical College, Huazhong University of Science & Technology from January 2014 to September 2019 were retrospectively reviewed.Results:The 41 patients included 23 males and 18 females with the onset age of (4.4±3.2) years.The main prodromal symptoms were gastrointestinal (20/41 cases, 48.8%) and respiratory infections (19/41 cases, 46.3%). Acute encephalopathy progressed rapidly following the prodromal infection [29 cases (70.7%) ≤2 days], and patients had clinical manifestations of coma (32/41 cases, 78.0%), convulsion (32/41 cases, 78.0%), multiple organ dysfunction (27/41 cases, 65.9%), shock and disseminated intravascular coagulation were rarely occured, and 28 cases (68.3%) were admitted to intensive care unit for treatment.Brain magnetic resonance imaging (MRI) showed lesion involving thalamus (41/41 cases, 100.0%), periventricular white matter (34/41 cases, 82.9%), brainstem (31/41 cases, 75.6%), basal ganglia (26/41 cases, 63.4%), cerebral cortex and subcortex (20/41 cases, 48.8%) and cerebellum (18/41 cases, 43.9%). The common presentations on the apparent diffusion coefficient mapping of brain MRI were " tricolor pattern" or " bicolor pattern" of the thalamus.During follow-up (≥ 6 months), MRI showed that hemorrhage, cystic degeneration and atrophy changed dynamically with the progression of ANE.All cases were treated with glucocorticoids, 38 cases(92.7%) with intravenous immune globulin.Seven cases (17.1%) were died and the 34 survivors had different degrees of neurological dysfunction.Conclusions:ANE in children is a distinctive type of clinicoradiologic syndrome with rapid progression and various presentations.Brain MRI has typical imaging characteristics and dynamically indicates the progression of this disease.The treatment options are still limited, the prognosis is poor and the survivors are often with neurological dysfunction.