OBJECTIVE To establish the characteristic chromatogram of Chaenomeles sinensis， determine the contents of rutin， hyperin and quercitrin， and to identify C. sinensis and C. speciosa. METHODS HPLC method was performed on Agilent 5 TC-C18 column， with acetonitrile-0.2% formic acid solution as the mobile phase for gradient elution， at the flow rate of 1.0 mL/min. The column temperature was 30 ℃ . The detection wavelength was 330 nm in characteristic chromatogram and 350 nm in content determination. The characteristic chromatogram of C. sinensis was established and similarity was evaluated by the Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 edition）. Hierarchical cluster analysis of 15 batches of C. sinensis （S1-S15） was performed by using SPSS 23.0 software. The contents of 3 flavones in 15 batches of C. sinensis and 7 batches of C. speciosa （S16-S22） were determined， while their characteristic chromatograms were compared. RESULTS The similarities of the characteristic chromatogram for 15 batches of C. sinensis ranged from 0.783 to 0.969， and 11 characteristic peaks were confirmed. Four constituents were identified as chlorogenic acid， rutin， hyperin and quercitrin. The medicinal materials in 15 batches of C. sinensis could be divided into 2 categories： S5-S8 were one category， and the others belonged to one category. The characteristic chromatogram of C. sinensis was obviously different from C. speciosa. The contents of rutin， hyperin and quercitrin in 15 batches of C. sinensis were 48.99-294.45， 3.49-102.55， 31.98-149.49 μg/g， respectively. The content of rutin in C. speciosa was lower than that in C. sinensis. None of hyperin （except for S20） and quercitrin were detected in C. speciosa. CONCLUSIONS The characteristic chromatogram and the method for content determination of 3 flavones in C. sinensis are established successfully and can be used for the quality control of C. sinensis and its identification from C. speciosa.

Objective To screen differentially-expressed circRNA in plasma exosomes of patients with non-small cell lung cancer(NSCLC)and verify its diagnostic value for NSCLC.Methods The plasma exosomes of six patients with NSCLC and six healthy people were analyzed by circRNA sequencing.The expression of hsa_circ_0052513 in plasma exosomes of 60 NSCLC patients and 60 healthy controls was detected by qRT-PCR.The expression level of hsa_circ_0052513 in plasma exosomes of NSCLC patients before and after surgery was detected by qRT-PCR.The correlation between the expression of plasma exosomal hsa_circ_0052513 and clinical data of patients with NSCLC was statistically analyzed.Results Sequencing results showed that the expression of hsa_circ_0052513 in the plasma exosomes of NSCLC patients was higher than that of healthy controls,and the results were confirmed by qRT-PCR.Hsa_circ_0052513 was decreased in patients with NSCLC after tumor resection(P<0.05).The high expression of plasma exosomal hsa_circ_0052513 in patients with NSCLC was correlated with tumor size,distant metastasis,and TNM stage of NSCLC(all P<0.05).The area under the curve of plasma exosomal hsa_circ_0052513 in the diagnosis of patients with NSCLC was 0.7904(P<0.0001).Conclusion Hsa_circ_0052513 is highly expressed in the plasma exosomes of patients with NSCLC and is related to tumor size,metastasis,and TNM stage.Hence,Hsa_circ_0052513 could be a new diagnostic marker for NSCLC.

OBJECTIVE: To explore the pathogenic variants and clinical classification of two fetuses with Short-rib thoracic dysplasia with or without polydactyly (SRTD). METHODS: With informed consent obtained, the phenotypic characteristics of the fetuses were comprehensively examined, and genomic DNA was extracted from fetal skin tissue and peripheral blood samples of the parents with conventional phenol-chloroform method. Whole exome sequencing (WES) was carried out on both fetuses, and the candidate variants were validated by Sanger sequencing. The pathogenicity of the candidate variants was analyzed using bioinformatic software VarCards, and the impact of the variants on the protein structure was predicted with Swiss-Pdb-viewer. RESULTS: Both fetuses were found to harbor compound heterozygous variants of the DYNC2H1 gene, including c.515C>A (p.Pro172Gln) and c.5983G>A (p.Ala1995Thr) in fetus 1, and c.5920G>T (pGly1974) and c.9908T>C (p.He3303Thr) in fetus 2. The parents of both fetuses were heterozygous carriers. CONCLUSION The compound heterozygous variants of the DYNC2H1 gene probably underlay the SRTD3 in the two fetuses.
Humans ; Fetus ; Chloroform ; Computational Biology ; Ethnicity ; Ribs

Objective:To explore the risk factors and follow-up outcomes of pediatric heart transplantation(HT).Methods:Between January 2018 and June 2022, perioperative data are retrospectively reviewed for 41 pediatric HT recipients aged <18 years and donor-recipient weight data for infants aged under 3 years at Guangdong Provincial People's Hospital.Perioperative survivors are followed up until August 31, 2022 through out patient visits and telephone calls.Postoperative survivals are examined by Kaplan-Meier method and possible risk factors for perioperative survival identify with Logistic regression.Results:There are 22 boys and 19 girls with a median age of 120(58~138)months.After preoperative adjuvant therapy of extracorporeal membrane oxygenation(ECMO), 8 cases had a successful transition to HT and 2 children underwent ABO incompatible(ABOi)HT.Six children aged under 3 years had a donor-recipient weight ratio of 2.95.Among 17 children, there are one or more complications, including continuous renal replacement therapy(CRRT, 9 cases, 21.95%), tracheotomy (3 cases, 7.32%), delayed chest closure or redo of sternotomy(6 cases, 14.63%)and acute graft dysfunction(4 cases, 9.76%). Five children died during perioperative period.The possible risk factors for perioperative mortality include preoperative ECMO assistance[ HR: 32.00, 95% CI: (2.83~361.79), P<0.05], preoperative CRRT[ HR: 11.33, 95% CI: (1.15~111.69), P<0.05] and total bilirubin [ HR: 1.02, 95% CI: (1.002~1.040), P<0.05]. During follow-ups, one child died from Epstein-Barr virus (EBV)associated post-transplant lymphoproliferative disease; another case of EBV-associated hepatic leiomyoma underwent transcatheter arterial embolization.With an overall survival rate of 85.37%, the cumulative survival rate is 96.97% for children without preoperative ECMO assistance( P<0.05). Postoperative mortality rate spiked markedly in children with preoperative ECMO assistance ( P=0.0013). However, follow-up results of perioperatively survivors indicate that preoperative usage of ECMO will not affect follow-up survival( P=0.53). In ABOi group or infants aged under 3 years, no mortality occurres postoperatively or during follow-ups. Conclusions:In infant aged under 3 years, the strategies of ABOi HT and large-weight donor HT are both safe and effective and it has no effect upon perioperative and follow-up survivals.Preoperative ECMO assistance, total bilirubin and preoperative use of CRRT are risk factors for perioperative survival.

Objective    To explore the correlation between perioperative blood transfusion and acute kidney injury (AKI) after heart transplantation. Methods    A retrospective study was performed on 67 patients who underwent heart transplantation in the Department of Cardiac Surgery, Guangdong Provincial People's Hospital from January 2016 to December 2018, and finally 63 patients were included according to the exclusion criteria. There were 53 males and 10 females with an average age of 44.3±12.9 years. Twenty patients who adopted continuous renal replacement therapy (CRRT) after heart transplantation were divided into a RT group and the other 43 patients who did not use CRRT were divided into a non-RT group. Baseline characteristics, perioperative blood transfusion data and clinical prognosis were compared between the two groups. Results    The preoperative baseline characteristics of the two groups were basically the same. There were significant differences in perioperative infusion of red blood cells and plasma, postoperative 24 h bleeding and re-exploration (P<0.05) between the two groups. The area under the receiver operating characteristic (ROC) curve was 0.923 (95%CI 0.852 to 0.995, P<0.001). The ROC curve showed that perioperative infusion of red blood cells more than 18 mL/kg would increase the incidence of AKI after heart transplantation. Conclusion    Perioperative blood transfusion is closely related to AKI after heart transplantation. The more blood transfusion is in clinics, the higher incidence of renal injury is and the worse prognosis is. It is suggested that various blood-saving measures can be carried out.

Objective:This study aims to evaluate the prognostic effect of peripheral blood cells in multiple myeloma (MM) patients treated with bortezomib.Methods:The clinical data of 155 newly diagnosed MM patients in two blood disease treatment centers from January 2014 to December 2016 were retrospectively studied. All patients received bortezomib as the first-line treatment. The results of the peripheral blood cell counts, including absolute neutrophil count, absolute monocyte count (AMC) , hemoglobin level, mean corpuscular volume (MCV) , and platelet count, and other clinical features were analyzed.Results:AMC (>0.6×10 9/L) , MCV (>99.1 fl) , and platelet count (<150×10 9/L) significantly affected patients’ PFS and OS. The above three factors were assigned 1 point, respectively, to form the blood cell score. The analysis showed that 64 cases (41.3% ) had a score of 0, 57 cases (36.8% ) had 1, 32 cases (20.6% ) had 2, and 2 cases (1.3% ) had 3. The median PFS of the four groups were 42.8 m, 26.5 m, 15.8 m, and 6.4 m, respectively ( P<0.001) . The median OS were NR, 48.2 m, 31.1 m, and 31.4 m, respectively ( P=0.001) . Multivariate analysis suggested that the blood cell score (2-3 vs 0-1) and the proportion of marrow plasma cells (>30% ) were independent prognostic factors for PFS ( HR=1.95 and 1.76, respectively) , while age (>65y vs ≤65y) , R-ISS stage (3 vs 1-2) , and blood cell score (2-3 vs 0-1) were independent prognostic factors for OS ( HR=2.08, 2.13 and 2.12, respectively) . Conclusion:As an easy-to-access biomarker, the blood cell score can be used to evaluate the prognosis of newly diagnosed MM patients in the era of new drugs, but it is still necessary to expand the cases and make further confirmation in the prospective study.

Objective To evaluate the effects of sodium-glucose cotransporter 2 ( SGLT2) inhibitors on the stroke risk for patients with type 2 diabetes mellitus (T2DM). Methods A systemic meta-analysis including 30 random control trails ( RCTs) was performed to compare the risk of stroke between type 2 diabetic patients treated with SGLT2 inhibitors and control drugs. Then their bias risk and quality were assessed and meta-analysis was conducted using Stata12.0 software. Results Thirty RCTs enrolling 74456 participants were selected for meta-analysis. The stroke incidence in the group receiving SGLT2 inhibitor monotherapy or combination therapy did not significantly differ from that in control group, with relative risk (RR) 1.01 (95％CI 0.93-1.10, P=0.978) and 1.00 (95％CI 0.92-1.09, P=0. 874 ) , respectively. Three SGLT2 inhibitors canagliflozin, dapagliflozin, and empagliflozin did not increase the risk of stroke, with similar RR values ( RR=0.91, 0.99, 1.13, respectively) . Subgroup analyses showed that there was no correlation between SGLT2 inhibitors and stroke risk in different gender, age, diabetes duration, body mass index, or HbA1C levels. Conclusions Whether administered as monotherapy or add-on therapy, SGLT2 inhibitors did not increase stroke incidence, and there were no significant within-class differences.

Objective: To study the effect of WT1 expression on the prognosis of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in acute leukemia (AL) and its significance as molecular marker to dynamically monitor minimal residual disease (MRD) . Methods: Retrospectively analyzed those AL patients who underwent allo-HSCT in the First Hospital Affiliated to Zhejiang University School of Medicine during Jan 2016 to Dec 2017, a total number of 314 cases, 163 males and 151 females, median age was 30 (9-64) years old. Comparing the difference of WT1 expression at diagnosed, pre-HSCT and after HSCT. Using the receiver operating characteristic (ROC) curve to determine the WT1 threshold at different time so as to predict relapse. The threshold of WT1 expression before transplantation was 1.010%, within 3 months after HSCT was 0.079% and 6 months after HSCT was 0.375%. According to these thresholds, WT1 positive patients were divided into low expression groups and high expression groups. Analyzed the relationship between overall survival (OS) , disease-free survival (DFS) , cumulative incidence of relapse (CIR) and WT1 expression. Results: The OS and DFS of high expression group pre-HSCT were lower than low expression group [69.2% (9/13) vs 89.1% (57/64) , χ²=4.086, P=0.043; 53.8% (7/13) vs 87.5% (56/64) , χ²=9.766, P=0.002], CIR was higher than low expression group [30.8% (4/13) vs 7.8% (5/64) , P=0.017]. There was no significant difference of OS and DFS between high expression and low expression group of 3 months after HSCT (P=0.558, P=0.269) . The OS and DFS of high expression group of 6 months after transplantation were both lower than low expression group (P=0.049, P=0.035) . Multivariate analysis showed that WT1>0.375% when 6 months after transplantation was the only independent prognostic factor for shorter DFS (P=0.022) . There was no statistically significant difference in CIR between the high-expression group and the low-expression group 3 months after transplantation and 6 months after transplantation (P=0.114, P=0.306) . Conclusion High expression of WT1 before and after HSCT was an adverse prognosis factor. It is of clinical practical value to use WT1 as a transplant recommendation index for patients with acute leukemia and as a marker to monitor MRD dynamically.

Objective: To evaluate the efficacy and safety of maintenance therapy with reduced dose of rhTPO in the patients with primary immune thrombocytopenia (ITP) who attained stable platelet (PLT) counts after daily administration of rhTPO. Methods: Treatment was started with a daily administration of rhTPO (300 U/kg) for 2 consecutive weeks. Patients who attained stable PLT≥50×10⁹/L were enrolled to maintenance therapy starting with every other day administration of rhTPO, then adjusted dose interval to maintain platelet count (30-100) ×10⁹/L. Results: A total of 91 eligible patients were enrolled. Fourteen patients discontinued the study due to noncompliance (12/14) and investigator decision (2/14) . Among 77 patients who completed the study, 38 patients with the administration of rhTPO at every other day or less could maintain PLT≥30×10⁹/L for 12 weeks. The percentage of patients with a platelet response (PLT≥30×10⁹/L) at 4^th week, 8^th week and 12^th week of maintain therapy was 92.6% (63/68) , 82.7% (43/52) and 85.0% (34/40) , respectively. Median platelet counts remained in the range of (70-124) ×10⁹/L. The overall incidence of rhTPO-related adverse events was 7.7%. All the adverse events were generally mild. Conclusion Extending the dose interval of rhTPO is feasible to maintain stable platelet count in the patients with ITP, but the optimal dose interval is uncertain and might vary with individuals.

Objective To evaluate clinical effectiveness of transcatheter closure of atrial septal defects ( ASD) with severe pulmonary arterial hyperyension ( sPAH) by fenestrated Amplatzer septal occluders ( ASO) . Methods From September 2002 to April 2013, 17 patients of ASD with sPAH received transcatheter ASD closure using fenestrated occluders. Aged from 18 - 72 years, the diameters of ASDs were 18 - 33 mm. The systolic pulmonary arterial hypertension measured by transthoracic echocardiogram were 80 - 112 (96. 9 ± 8. 9) mmHg. The follow-up study included electrocardiography, chest radiography and echocardiography. All the patients were followed up for 1. 5 - 12 ( mean 6. 4 ± 0. 7) years. Results Systolic pulmonary arterial pressure (sPAP) of 60 - 108 (88. 7 ± 11. 7) mmHg and mean pulmonary artery pressure ( mPAP) of 29. 3 - 60 (51. 0 ± 8. 1) mmHg were measured by cardiac catheterization before ASD closure. Qp/ Qs was 1. 50 - 2. 44 (1. 8 ± 0. 31) and the pulmonary vascular resistance was 3. 1 - 9. 7 (5. 6 ± 1. 5) wood units (wu) . Immediately after the implantation of fenestrated occluders, sPAP decreased to 56 - 99 (70 ± 11. 5) mmHg and mPAP to 27 - 51. 7 (41. 1 ± 7. 1) mmHg. On the 3 d, 3 m and 6 m follow-up exam, RVEDd decreased ( P ﹤ 0. 05), while LVEDd, LVEDV and LVEF increased significantly (P ﹤ 0. 05) . sPAP decreased significantly after transcatheter closure at 3 m and 6 m as compared to pre-closure levels (both P ﹤ 0. 05) . The mean sPAP in long term follow up was (60. 2 ± 13. 3) mmHg which had significant decrease compared to pre-closure level ( P ﹤ 0. 01), but no significant difference found when compared to 6 m follow up (P ﹥ 0. 05). Conclusions ASD closure with fenestrated ASO is a satisfactory approach for ASD with severe PAH.