[Study on the inhibitory and pro-apoptotic effects of different concentrations of total tanshinone alone and in combination with tyrosine kinase inhibitors on human myeloid leukemia cell lines].

Objective: To explore the effect and investigate the molecular mechanism of different concentrations of total tanshinones alone and in combination with tyrosine kinase inhibitors (TKIs) on the proliferation inhibition and apoptosis of human myeloid leukemia cell lines. Methods: K562 and Kasumi-1 cell lines were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences, and the TKIs-resistant strain K562/T315I cell line was constructed in Molecular Medicine Research Center, Beijing Lu Daopei Institute of Hematology. Logarithmic growth phase cells were taken and divided into intervention groups with total tanshinone of 0, 2.19, 4.38, 8.75, 17.50 and 35.00 µg/ml intervention groups, which were inoculated in 96-well plates at a density of 1×104 cells/well and exposed to the drug for 24 h, and a control group treated with dimethyl sulfoxide was also set up simultaneously. All experiments were repeated independently 3-5 times. The proliferative activity of the cells was assessed using the CCK-8 assay, the apoptotic rates were measured by flow cytometry, and the expression levels of apoptosis-regulating proteins Bcl-2 and Bax were analyzed by Western blotting. The cell lines treated and untreated with total tanshinone were subjected to transcriptome sequencing and gene set enrichment analysis to identify differentially expressed genes. Results: The half-inhibitory concentration (IC50) values of 8.75 µg/ml total tanshinone at 24 h for K562, K562/T315I and Kasumi-1 cells were (4.11±0.02), (4.95±0.04) and (3.98±0.01) µg/ml, respectively. When combined with 0.25 µmol/L imatinib, 8.75 µg/ml total tanshinone could enhance the induction of apoptosis effects on K562 and K562/T315I cell lines. After being treated with 4.38, 8.75, and 17.50 µg/ml of total tanshinone for 24 h, compared with the control group, total tanshinone upregulated the expression level of Bax protein, downregulated the expression level of Bcl-2 protein, and decreased the Bcl-2/Bax ratio (all P<0.05). Total tanshinone inhibited the proliferation-related signaling pathway and DNA damage repair pathway of myeloid leukemia cell lines, and activated the signaling pathway that induces apoptosis in leukemia cells. Conclusion: Different concentrations of total tanshinoneinhibites proliferation and promote apoptosis in K562, Kasumi-1 and TKIs-resistant K562/T315I cell lines, and further enhance the anti-leukemic effect when combined with TKIs.

[Correlation between common driver gene variations and clinicopathological typing in lung adenocarcinoma].

Objective: To correlate the common driver gene variations in primary lung adenocarcinoma with their clinical characteristics and histopathological subtypes. Methods: There were 4 995 cases of primary lung adenocarcinoma diagnosed at Weifang People's Hospital of Shandong Province from January 2015 to December 2021 which were retrospectively analyzed. Among them 1 983 cases were evaluated for their histopathological subtype; 3 012 were analyzed for the correlation of their histopathological subtypes and corresponding driver gene variations, including invasive non-mucinous adenocarcinoma (INMA) and invasive mucinous adenocarcinoma (IMA), and morphologically, poorly-differentiated, moderately-differentiated and well-differentiated adenocarcinomas. Next-generation sequencing was used to detect variations in EGFR, KRAS, ALK, RET, ROS1, MET, HER2, or BRAF driver genes. Results: There were 2 384 males and 2 611 females. EGFR and ALK variations were more commonly found in female patients aged 60 years or older, with EGFR mutation rate in clinical stage Ⅰ (25.80%) significantly higher than in other stages (P<0.05). KRAS mutations were more commonly detected in male smokers aged 60 years or older, HER2 mutations were more commonly in patients younger than 60 years, and RET mutations were more commonly in non-smokers (all P<0.05). No correlation was found between ROS1, MET, and BRAF gene variations and their clinical characteristics (P>0.05). For the histopathological subtypes, among the 1 899 cases of acinar adenocarcinoma, EGFR mutation rate was the highest (67.30%) compared to the other genes. Exon 21 L858R and exon 19 del were the main mutation sites in IMA and INMA, with a higher mutation rate at exon 20 T790M (11.63%) in micropapillary adenocarcinoma. In IMA, KRAS had the highest overall mutation rate (43.80%), with statistically significant difference in mutation rates of exon 2 G12D and exon 2 G12V in acinar adenocarcinoma, solid, and IMA (P<0.05). KRAS mutation at various sites were higher in poorly differentiated groups compared to moderately- and well-differentiated groups (P<0.05). HER2 mutations were more commonly observed in acinar adenocarcinoma, papillary, and micropapillary adenocarcinoma of INMA. BRAF mutation was higher in micropapillary adenocarcinoma compared with other types (P<0.05). Conclusions: Variations in EGFR, ALK, KRAS, HER2, and RET in primary lung adenocarcinoma are associated with patients' age, smoking history, and clinical stage, and driver gene mutations vary among different histopathological subtypes. EGFR mutations are predominant in INMA, while KRAS mutations are predominant in IMA.

[Comparison of quantitative detection of BCR::ABL1 p210 transcript levels: a multicenter study].

Objective: To assess the capability of seven reference medical laboratories to detect BCR::ABL1 p210 transcription levels and to compare the results among those laboratories. Methods: The interlaboratory comparison was carried out in two stages. The samples were prepared by the reference laboratory. The quantitative values of BCR::ABL1 p210 of the comparison samples covered 0.001%-0.01%, 0.01%-0.1%, 0.1%-1%, 1%-10% and>10% in each stage. Real-time quantitative PCR (RT-PCR) and dPCR (digital PCR) were used to examine the samples. The conversion factor (CF) was calculated and validated for each laboratory. Results: In the RT-PCR comparison, one laboratory was failed to detect BCR::ABL1 p210 in fourteen samples at the first stage. The results of the other six laboratories were qualified with the bias <±1.2 folds (-0.133-0.338) and 95% limits of agreement within ±5 folds (upper limit 0.147-0.785, lower limit -0.770--0.109), and the corresponding CF values were calculated and validated. In the dPCR comparison, one laboratory did not report results at the second stage. The results of the other six laboratories were qualified with the bias <±1.2 folds (-0.026-0.267) and 95% limits of agreement within±5 folds (upper limit 0.084-0.991, lower limit -0.669--0.135), and the corresponding CF values were calculated and validated. The samples with BCR::ABL1 p210 quantitative values of 0.01%-0.1%, 0.1%-1%, 1%-10% and >10% could be detected by both RT-PCR and qPCR. When the quantitative value of BCR::ABL1 p210 was 0.001%-0.01%, the detection rate of dPCR was higher than that of RT-PCR (85.56% vs. 68.00%). Conclusions: A good consistency is present among various laboratories. The quantitative value of BCR::ABL1 p210 is comparable among laboratories as shown by the CF value conversion. For quantitative detection of BCR::ABL1 p210 deep molecular reaction, dPCR has a higher positive detection rate and more advantages than RT-PCR. To ensure the accuracy and reproducibility of the BCR::ABL1 p210 test, it is imperative for every laboratory to enhance their daily quality control practices.

[The clinical value of serum GPC3 level in predicting recurrence of patients with primary hepatocellular carcinoma].

Objective: To investigate the clinical value of serum glypican-3 (GPC3) detection in predicting recurrence of primary hepatocellular carcinoma (HCC). Methods: Through univariate and multivariate logistic regression analysis, the patients pathologically diagnosed with HCC in our hospital from March 2019 to January 2021 were enrolled as the experimental group (n=113), and patients with follow-up time longer than 6 months were included in the prognosis group(n=64). At the same time,20 healthy individuals and 20 individuals with benign liver disease from the physical examination center were enrolled by simple random sampling as control group (n=40). The serum GPC3 and alpha-fetoprotein (AFP) levels were respectively detected by ELISA and chemiluminescence. Then, the study explored the influential factors of the recurrence in HCC patients and constructed the HCC-GPC3 recurrence predicting model by logistic regression. Results: In the research, the sensitivity of GPC3 for the diagnosis of HCC was 61.95% (70/113) and AFP was 52.21% (59/113), meanwhile, the specificity of GPC3 could reach 87.50% (35/40) and AFP was 90.00% (36/40),respectively; The serum GPC3 levels of HCC patients with progressive stage, tumor size≥3 cm, vascular cancer thrombosis and portal venous thromboembolism were significantly higher than that of HCC patients with early stage, tumor size<3 cm, vascular cancer thrombosis and portal venous thromboembolism (Z=2.677, 2.848, 2.995, 2.252, P<0.05), independent of different ages, presence or absence of ascites, peritoneal metastasis, cirrhosis, intrahepatic metastasis (Z=-1.535, 1.011, 0.963, 0.394, 1.510, P>0.05), respectively. Univariate analysis showed that there were no statistically significant differences between the recurrence group and the non-recurrence group in terms of different age, tumor size, presence or absence of vascular cancer thrombosis, ascites, peritoneal metastasis, cirrhosis and AFP levels (χ2=2.012, 0.119, 2.363, 1.041, 0.318, 0.360, Z=0.748, P>0.05); The ratio of those with the progressive stage, portal venous thromboembolism and intrahepatic metastasis and GPC3 levels were all higher in the recurrence group than in the non-recurrence group (χ2=4.338, 11.90, 4.338, Z=2.805, P<0.05).Including the above risk factors in the logistic regression model, the logistic regression analysis showed that the stage, the presence of portal venous thromboembolism,intrahepatic metastasis and GPC3 levels were correlated with the prognosis recurrence of HCC patients (Wald χ2=4.421, 5.681, 4.995, 4.319, P<0.05), and the HCC-GPC3 recurrence model was obtained as: OcScore=-2.858+1.563×[stage]+1.664×[intrahepatic metastasis]+2.942×[ portal venous thromboembolism]+0.776×[GPC3]. According to the receiver operating characteristic curve(ROC), the area under the curve(AUC)of the HCC-GPC3 prognostic model was 0.862, which was better than that of GPC3 alone (AUC=0.704). The cut-off value of model SCORE was 0.699 (the cut-off value of GPC3 was 0.257 mg/L), furthermore, the total sensitivity and specificity of model were 83.3% and 82.4%, which were better than those of GPC3(60.0% and 79.4%).Kaplan-Meier showed that the median PFS was significantly shorter in HCC patients with high GPC3 levels (≥0.257 mg/L) and high values of the model SCORE (≥0.700) (χ2=12.73, 28.16, P<0.05). Conclusion: Besides diagnosing of HCC, GPC3 can may be an independent risk indicator for the recurrence of HCC and can more efficiently predicting the recurrence of HCC patients when combined with the stage, the presence or absence of intrahepatic metastasis and portal venous thromboembolism.

[Efficacy and prognostic analysis of 125I brathytherapy combined with chemotherapy for pediatric parameningeal rhabdomyosarcoma].

Objective: To analyze the efficacy and prognostic factors in children with parameningeal rhabdomyosarcoma (PMRMS) treated by 125I brachytherapy combined with chemotherapy. Methods: A retrospective analysis of the clinical data of 33 pediatric patients treated with 125I brachytherapy combined with chemotherapy in Peking University Stomatological Hospital from July 2013 to October 2018 was carried out to analyze the efficacy and prognostic factors. Results: Among the 33 patients, 19 were males and 14 were females; the median age was 4 years old (1-12 years old). There were 17 cases with embryonic type, 9 cases with alveolar type, and 7 with undifferentiated type; 26 cases with original PMRMS, and 7 cases with recurrent PMRMS. The tumors occurred in subtemporal-mastoid area in 15 patients, while nasopalatine-paranasal area in 6 cases, and parapharyngeal-submandibular area in 12 cases. There were 28 patients in IRS Ⅲ, and 5 patients in IRS Ⅳ. As for the risk level, 28 cases were in the middle-risk group and 5 cases in the high-risk group. The median follow-up time was 52 months. The 1, 3, and 5-year local control rates were 87.9%, 58.6%, and 49.9%, and the 1, 3, and 5-year survival rates were 93.8%, 60.5%, and 47.5%, respectively. The 5-year local control rate and 5-year survival rate of 12 patients with the tumor in the parapharyngeal-submandibular area were 91.7% and 100%, respectively. The 5-year local control rate and 5-year survival rate of the 6 patients with tumor in the nasopalatine-paranasal area were both 83.3%. The 3-year local control rate and 3-year survival rate of the 15 patients with tumor in the subtemporal-mastoid area were 17.5% and 21.4%. The multivariate survival analysis using Cox proportional risk regression model showed that the tumor located in the subtemporal-mastoid area was an independent risk factor affecting the 5-year overall survival rate (HR=38.40, 95%CI: 4.87-302.52, P=0.001). Within 3 months after 125I seed implantation, the incidence of acute radiotherapy adverse reactions in all patients was 84.8% (28/33). Twenty-one patients (63.6%) had a grade 1 acute radiotherapy reaction, and 7 cases (21.2%) had a grade 2 acute radiotherapy reaction. No acute radiotherapy adverse reactions of grade 3 or 4 occurred. Three months after 125I seed implantation, the adverse reactions were significantly alleviated, and no adverse reactions of grade 3 or above such as skin ulcer or salivary gland fibrosis occurred, and no serious cranio-maxillofacial deformities occurred. Conclusions: 125I seed brachytherapy combined with chemotherapy has a definite clinical effect in the treatment of children with parameningeal rhabdomyosarcoma. The prognosis of rhabdomyosarcoma in the parapharyngeal-submandibular area and nasopalatine-paranasal area is better than that in the subtemporal-mastoid area.

Polar meron lattice in strained oxide ferroelectrics.

A topological meron features a non-coplanar structure, whose order parameters in the core region are perpendicular to those near the perimeter. A meron is half of a skyrmion, and both have potential applications for information carrying and storage. Although merons and skyrmions in ferromagnetic materials can be readily obtained via inter-spin interactions, their behaviour and even existence in ferroelectric materials are still elusive. Here we observe using electron microscopy not only the atomic morphology of merons with a topological charge of 1/2, but also a periodic meron lattice in ultrathin PbTiO3 films under tensile epitaxial strain on a SmScO3 substrate. Phase-field simulations rationalize the formation of merons for which an epitaxial strain, as a single alterable parameter, plays a critical role in the coupling of lattice and charge. This study suggests that by engineering strain at the nanoscale it should be possible to fabricate topological polar textures, which in turn could facilitate the development of nanoscale ferroelectric devices.

[Spatial structure of talus trabeculae based on high resolution X-ray and micro-CT].

Objective: To study the specific alignment and structure of cancellous bone within the talus in order to understand the mechanism of force transmission within the bone and to provide some theoretical basis for the repositioning of talar fractures and the design of prostheses. Methods: In January 2020, a total of 40 adult talar bone specimens were scanned by Micro-CT in 20 pairs obtained from the Department of Orthopedics of Tianjin Hospital. The bone volume fraction, bone surface area fraction, trabecular thickness, number of trabeculae, trabecular pattern factor of the head, neck and body of the talus were calculated, and the differences in each parameter were compared between different parts of the same side and different sides of the same part, respectively. The talus was cut into 2 mm thick slices in the coronal, sagittal and horizontal planes using a hard tissue slicer, and the slices were then scanned using high-resolution X-rays to describe the bone structure. Results: There were no statistically significant differences between the medial and lateral talar and right and left side in lateral trabecular bone volume fraction, bone surface area fraction, trabecular thickness, trabecular number, trabecular pattern factors (all P>0.05). The number of trabeculae in the talar head, neck and body was 1.608±0.150, 1.639±0.142 and 1.749±0.159, respectively; trabecular thickness (µm) in the talar head, neck and body was 0.378±0.054, 0.370±0.053 and 0.331±0.062, respectively; and the trabecular pattern factors (mm-1) in the talar head, neck and body was -0.407±0.699, -0.478±0.848 and -1.029±0.851, respectively. There were significant differences between talar head, neck and the talar body trabeculae in terms of the number of trabeculae, trabecular thickness,trabecular pattern factor parameters(all P<0.05). The structure of the talar body trabeculae was found to consist of plate trabeculae arranged vertically parallel to each other in the coronal, sagittal and horizontal planes. The talar neck trabeculae were twisted, external-superior to internal-inferior reticular plate structure that travelled posteriorly and anteriorly, and the talar head trabeculae consisted of similarly parallel aligned semi-arc-shaped external-superior and internal-inferior trabeculae. Conclusion: The talar trabeculae are clearly directional and functional, so anatomical reduction should be achieved after the fracture; at the same time, the design of the talar prosthesis should take into account the stress distribution and direction of the prosthesis during walking and standing.

[Transcriptome sequencing-based classification and quantification of IKZF1 transcript isoforms in B-cell acute lymphoblastic leukemia].

Objective: To classify and quantify IKZF1 mutant transcripts in B-cell acute lymphoblastic leukemia (B-ALL) by RNA sequencing (RNA-seq) and bioinformatics analysis. Methods: A cohort of 263 B-ALL cases was enrolled at Hebei Yanda Ludaopei Hospital from September 2018 to September 2020. An integrated bioinformatics pipeline was developed to adapt the classification and quantification of IKZF1 transcripts from RNA-seq and was applied to sequencing data of these cases. The IKZF1 mutant transcripts classified by RNA-seq analysis were compared with the qualitative reverse transcription PCR (RT-PCR). Results: IKZF1 mutant transcripts were identified in 53 B-ALL patients by RT-PCR and Sanger sequencing, among which IK6 and IK10 transcripts accounted for 67.9% (36/53) and 28.3% (15/53) respectively. Additionally, 2 patients were double positive for IK6 and IK10. RNA-seq analysis identified 51 patients with IKZF1 mutant transcripts. Compared with the RT-PCR result, the detection sensitivity and specificity of RNA-seq analysis reached 94.3% (50/53) and 99.5% (209/210), respectively. Among the 50 patients with IKZF1 mutant transcripts both in RNA-seq and RT-PCR analysis, the ratio of mutant transcripts to total IKZF1 transcripts in 6 patients was 0.14 (0.11, 0.35), which was significantly lower than that of the other 44 patients [0.88 (0.35, 0.97), Z=-3.945,P<0.001]. IKZF1 mutations mostly occurred in Ph+and Ph-like B-ALL, characterized by abnormal JAK-STAT pathway, and B-ALL with PAX5 translocation. Conclusions: Through the optimized bioinformatics analysis process, RNA-seq data can be used to classify and quantitatively analyze IKZF1 transcripts in B-ALL. Furthermore, the relative expression of mutant IKZF1 transcripts was found to cluster into two groups, and IKZF1 mutation was found often accompanied with PAX5 translocations.

[The impact of KIT and other concomitant gene mutations on the prognoses of patients with core-binding factor acute myeloid leukemia].

Objective: To study the impact of KIT and other concomitant gene mutations on the prognoses of patients with core-binding factor acute myeloid leukemia (CBF-AML). Methods: A total of 104 newly diagnosed patients with CBF-AML in Hebei Yanda Lu Daopei Hospital from January 2014 to February 2018 were analyzed, and high-throughput gene sequencing for the detection of mutations among 58 genes was executed. Also, the clinical features of KIT mutation-positive CBF-AML (KIT+CBF-AML) patients and the effects of other concomitant gene mutations on the prognoses of patients were also analyzed. Results: A total of 56 cases (53.85%) with KIT mutations were found in 104 CBF-AML patients. Among this, KIT D816 mutation was the most common (32 patients), followed by the N822 mutation (17 patients). Patients with KIT+CBF-AML have a higher proportion of bone marrow blasts at the time of diagnoses and are more likely to have sex chromosome loss. Among the 52 patients with KIT+CBF-AML who were followed up, the allogeneic hematopoietic stem cell transplantation (allo-HSCT) group had a higher overall survival rate (OS) than that of the chemotherapy group (88.9% vs 57.1%, χ(2)=6.076, P<0.05). The event-free survival (EFS) and OS of patients with KIT+CBF-AML with FLT3 mutation were both significantly lower than those of the FLT3 mutation-negative group (EFS: 40.0% vs 72.3%, χ(2)=6.557, P<0.05; OS: 60.0% vs 87.2%, χ(2)=8.305, P<0.05). The OS of the patient with TET2 mutation was lower than that of the TET2 mutation-negative group (50.0% vs 87.5%, χ(2)=4.130, P<0.05). Conclusion: Patients with KIT+CBF-AML with concomitant gene mutations, especially FLT3 and TET2, have poor prognoses, which can be improved by allo-HSCT.

[Short-term outcomes of sequential radial artery grafting in patients with coronary artery disease].

Objective: To explore the short-term outcomes of patients with coronary artery disease (CAD) who underwent coronary artery bypass graft (CABG) with sequential radial artery graft. Methods: Clinical data of patients with CAD who underwent CABG with sequential radial artery graft from August to December 2018 in Beijing Anzhen Hospital was retrospectively analyzed. Computer tomography angiography was employed to evaluate the patency of grafts and the incidence of adverse cardiovascular events was followed up 3 months after surgery. Results: A total of thirty patients were included, with an average age of (60.2±7.3) years old, among whom 83.3% were male, and 93.3% had three-vessel disease detected by coronary angiography. All patients had left internal mammary artery (LIMA) anastomosed to the left anterior descending (LAD) and sequential radial artery graft. Among sequential radial artery grafts, the most frequent anastomosis of sequential radial artery graft was aortic artery (AO)-first diagonal branch (D1)-first obtuse marginal branch (OM1) (13 patients, 43.3%), followed by AO-OM1-second obtuse marginal branch (OM2) (9 patients, 30.0%), and 18 patients also underwent saphenous vein graft. None of 30 patients experienced adverse cardiovascular events during postoperative period in hospital. Three-months follow-up outcomes showed that LIMA-LAD in 1 patient (3.3%) occluded, 3 patients had occluded anastomosis of sequential radial artery graft, 1 patient (3.3%) had heart failure, and 1 patient (3.3%) had recurrent angina. Conclusion: The radial artery could be used as the second arterial graft for CAD patients undergoing CABG with a sequential anastomosis technique, and these patients need regular anti-arterial spasm drug treatment after CABG.

[Quantitative study on the development of fetal ventricles and cisterna magna in second and third trimesters with MRI].

Objective: To analyze the changes of growth and development of normal fetal ventricles and cisterna magna with gestational age(GA) and the correlation with fetal gender in the second and third trimester,and establish the MR prenatal diagnosis reference standards. Methods: A total of 633 fetuses (mean GA (27.0±4.1) weeks (18.9-40.6 weeks))without central nervous system abnormalities were retrospectively collected from the Obstetrics and Gynecology Hospital of Fudan University from June 2012 to August 2017. The lateral ventricle trigonometric width (LVTW), third ventricle width (TVW), fourth ventricle width (FVW), anterior-posterior diameter of the fourth ventricle(APDFV), cavum septum pellucidum width (CSPW) and cisterna magna width (CMW) were obtained in the standard measure planes on MR image.The correlation between the biometrics and GA and the correlation between the biometrics and fetal gender were analyzed respectively, and the normal reference values of the biometrics were calculated. Spearman correlation analysis, Pearson correlation analysis,linear regression analysis, independent samples t-test and paired samples t-test were used for statistic analysis. Results: (1)Fetal LLVTW,RLVTW,TVW,CSPW and CMW in second and third trimesters were correlated with GA at medium and low levels(the correlation coefficient r were 0.311, 0.277, 0.207, 0.226, 0.295, respectively, all P<0.01). FVW and APDFV were statistically correlated with GA, and the linear regression equations were as follows: y=0.022×GA-0.043 (adjusted R(2)=0.642); y=0.018×GA-0.159 (adjusted R(2)=0.690). (2)Fetal LLVTW,RLVTW,FVW,APDFV and CSPW were not correlated with fetal gender in second and third trimesters(r=-0.078,-0.057,-0.087,-0.004 and 0.024, P=0.124,0.258,0.085,0.931 and 0.618, all P>0.05). TVW and CMW were statistically correlated with fetal gender(r=-0.310, -0.180, P=0.000, 0.006, all P<0.05). (3) The mean values of LLVTW and RLVTW were (0.71±0.13) cm and (0.68±0.13) cm, respectively, and significant difference was found between them(t=3.180, P=0.002). The mean value of CSPW was (0.59±0.15) cm. And the mean values of male and female fetuses for TVW and CMW were (0.17±0.05) cm, (0.16±0.06) cm and (0.68±0.15) cm, (0.58±0.15) cm, respectively. The corresponding prenatal MRI diagnostic criteria were as follows: LLVTW 1.1 cm, RLVTW 1.0 cm, CSPW 1.0 cm, TVW 0.3 cm, CMW (male 1.1 cm, female 1.0 cm). Conclusions: The normal fetal ventricles and cisterna magna are increased with the GA in the second and third trimesters. TVW and CMW are related to fetal gender. The establishment of normal reference values of fetal ventricles and cisterna magna based on GA and fetal gender are conducive to enhance the accuracy of MRI prenatal diagnosis.

[Genetic characteristics and clinical outcomes of pediatric acute myeloid leukemia with NUP98-NSD1 fusion gene].

Objective: To investigate the genetic characteristics and clinical outcomes of pediatric acute myeloid leukemia patients with NUP98-NSD1 fusion gene. Methods: A total of 80 pediatric AML patients were enrolled in this study, and bone marrow specimens were collected at initial diagnosis and relapse. NUP98-NSD1 was screened by fluorescence in situ hybridization (FISH) and PCR. Other laboratory test results and clinical outcomes were further analyzed for the NUP98-NSD1 positive cases. Results: A total of eight patients (10.0%) were positive for NUP98-NSD1, which were all fusions of NUP98 exon12 and NSD1 exon 6. There were two M2, three M4, and three M5 cases according to the French-American-British classification. Seven patients had karyotype results at the time of initial diagnosis, and none of them had complicated karyotype abnormalities. Among these patients, two cases had normal karyotype, three cases had trisomy 8, one case had trisomy 6, and two cases had anomalies involving 9q13 or 9q21. Additional karyotypic abnormalities and clonal evolutions were observed during disease progression or relapse, five cases had 9q13 or 9q32 abnormalities. Five cases (62.5%) were positive with FLT3-ITD mutation. Patients were treated with DAE/NAE/HAE/IA chemotherapy. Three cases did not achieve remission after several courses of chemotherapy, and five cases achieved remission but relapsed in 1 to 19 months. Five cases underwent salvage allogeneic hematopoietic stem cell transplantation (allo-HSCT). Among whom, four died in 40 days to 4 months after transplantation, and one survived 8.5 months till the last follow-up. Conclusions: NUP98-NSD1 is a recurrent genetic abnormality with significant clinical prognostic significance, and this group of disease has unique clinical and genetic characteristics. NUP98-NSD1 should be screened by FISH or PCR for children with AML who are newly diagnosed or refractory and relapsed to identify the high-risk genetic marker.

[Effectiveness evaluation between enhanced recovery after surgery and traditional treatment in unilateral total knee arthroplasty].

Objective: To investigate the clinical effect of enhanced recovery after surgery (ERAS) in unilateral total knee arthroplasty (TKA). Methods: Retrospective analysis of 98 patients received unilateral TKA from September 2015 to September 2016 in the Orthopaedic Departmentof Affiliated Hospital of Logistics University of Chinese People's Armed Police Forces.Of the patients, 52 cases were treated by traditional operation (routine group) and the other 46 cases were treated with ERAS concept (ERAS group). The following data were gathered and statistically analyzed between the groups: gender, age, body mass index, preoperative hemoglobin, preoperative visual analogue scale (VAS) score, postoperative visible blood loss, postoperative hemoglobin at 24 hour after operation, blood transfusion rate, postoperative VAS score, length of hospital stay, the satisfaction rate, preoperative and 1 and 6 months postoperative evaluation with hospital for special surgery knee score (HSS), postoperative range of motion (ROM) of knees after 2 weeks, 1 month and 6 months of surgery, incidence rate of complications after surgery.Data between groups were analyzed with one-factor analysis of variance or chi-square test. Results: There was no significant difference in preoperative data between two groups.Postoperative visible blood loss of ERAS group was significantly lower than that in routine group[(224±59) vs (361±70) ml, t=4.723, P<0.01]; postoperative hemoglobin after 24 hours in ERAS group was significantly higher than that in routine group[(109±8) vs (96±10) g/L, t=-3.297, P=0.004]; blood transfusion rate of ERAS group was significantly lower than that in routine group[6.5 %(3/46) vs 46.2%(24/52), χ(2)=19.207, P<0.01]; the 12-hour and 48-hour postoperative VAS scores in ERAS groups were both significantly lower than those in routine group (t=3.708, 3.894, both P<0.05); length of hospital stay in ERAS group was significantly shorter than that in routine group[(6.8±1.2) vs (13.1±2.6) d, t=6.924, P<0.01]; HSS scores of ERAS groups at 1 month and 6 months after surgery were both significantly higher than those in routine group (t=-3.677, -3.594, both P<0.05); knees ROM in ERAS group at 2 weeks and 1 month after surgery were both significantly higher than those in routine group (t=-4.628, -4.442, both P<0.05); the rate of postoperative nausea vomiting in ERAS group was significantly lower than that in routine group[13.0%(6/46) vs 48.1%(25/52), χ(2)=13.852, P=0.002]. VAS score at 1 month after surgery, knees ROM at 6 months after surgery and the satisfaction rate were all comparable between the two groups (t=0.412, -1.026, χ(2)=3.695, all P>0.05). Conclusions: Condition of patients treated by unilateral TKA under ERAS model improves effectively during perioperative period when compared with routine treatment, patients have earlier rehabilitation and better experience in hospital and operation.

[The application of computer-assisted design in the reduction of long bone fractures with Taylor spatial frame].

Objective: To evaluate the effect of computer-assisted design based on three-dimensional reconstruction technique on the reduction accuracy of tibial and fibular fractures with Taylor external fixation. Methods: A retrospective review was conducted on the clinical data of 69 patients who had tibia and fibula fractures treated with Taylor external fixation in department of orthopedic trauma of Tianjin Hospital from January 2016 to January 2018 to compare the residual deformity after fracture reduction between computer-assisted design method (experimental group) and the standard measurement method (control group). The frontal and lateral tibia and fibula X-ray of all the affected limbs were taken. In experimental group, all the patients took bilateral tibial CT tomography, and then DICOM format documents were input into the Mimics 17.1 software and got three-dimensional models of targeted bone and external fixation ring. After that the visual image matching was performed between external fixation ring three-dimensional reconstruction model and the standard model and also between the affected limb and the contralateral limb. Then the reduction trajectory plan of bone broken end and the position of external fixation ring were obtained. The STL files were input to Solid Works software and got the length of six rods to adjust the Taylor external fixation. In control group, the films were measured by Coreldraw X7 X-ray measurement software and the parameters were input in Taylor Spatial Frame system software. And then six calibrated threaded rods were adjusted according to the prescription of the software. Finally, all the patients took the X-ray films again to evaluate the degree of residual displacement. Skew distributional data are indicated with M(QR), and method of non-parameter was used to analyze variances between groups. Results: All patients had better fracture reduction and achieved functional reset criteria. In the control group, the amount of displacement and angle residual aberration (improvement) in the frontal and lateral radiographs were 0.50(2.30)mm(90%), 0.00(0.85)mm(100%)and 0.00°(1.50°)(100%), 0.00°(0.00°)(100%), respectively. In the control group, the amount of displacement and angle residual aberration (improvement) in the frontal and lateral radiographs were 1.40(3.28)mm(69%), 2.15(4.27)mm(46%)and 1.15°(1.85°)(73%), 0.80°(2.10°)(67%). The positive and lateral angles and lateral displacements in the two groups were significantly different(P<0.05), but there was no statistically significant difference in positive displacement (P=0.099). Conclusion: Both computer-assisted design method and the standard measurement method have satisfactory reduction effect, but computer-assisted design can accurately correct fracture deformity, which is good for fracture healing and functional recovery of affected limb.

[Long-term efficacy of open reduction and internal fixation versus external fixation for unstable distal radius fractures: a meta-analysis].

Objective: To make a systematic assessment of the Long-term efficacy of open reduction and internal fixation versus external fixation for unstable distal radius fractures. Methods: A computer-based online search of PubMed, ScienceDirect, EMBASE, BIOSIS, Springer and Cochrane Library were performed. The randomized and controlled trials of open reduction and internal fixation versus external fixation for unstable distal radius fractures were collected. The included trials were screened out strictly based on the criterion of inclusion and exclusion. The quality of included trials was evaluated. RevMan 5.0 was used for data analysis. Results: Sixteen studies involving 1 268 patients were included. There were 618 patients with open reduction and internal fixation and 650 with external fixation. The results of meta-analysis indicated that there were statistically significant differences with regard to the complications postoperatively (infection(I(2)=0%, RR=0.27, 95% CI 0.16-0.45, Z=4.92, P<0.000 01) and total complications(I(2)=0%, RR=0.71, 95% CI 0.59-0.85, Z=3.65, P=0.000 3) ), DASH scores(I(2)=37%, MD=-5.67, 95% CI -8.31--3.04, Z=4.22, P<0.000 1) and volar tilt(I(2)=78%, MD=2.29, 95% CI 0.33-4.24, Z=2.30, P=0.02)(P<0.05) at the end of follow-up period were noted. There were no statistically significant differences observed between two approaches with respect to the clinical outcomes (grip strength, flexion, extension, pronation, supination, radial deviation and ulnar deviation) and radiographic outcome(radial length) at the end of follow-up period(P<0.05). Conclusion: Both open reduction and internal fixation and external fixation are effective treatment for unstable distal radius fractures. Compared with external fixation, open reduction and internal fixation provides reduced complications postoperatively, lower DASH scores and better restoration of volar tilt for treatment of distal radius fractures.

[Efficiency of the Modified Ottawa ankle rules for the differential diagnosis of fracture in acute foot and ankle injury].

Objective: To evaluate the efficiency of modified Ottawa Ankle Rules (OAR) for the differential diagnosis of fractures in acute foot and ankle injuries. Methods: From October 2016 to December 2016, 272cases (135 males and 137 females) of foot and ankle injury in emergency department of Tianjin Hospital were prospective enrolled in the study.The median age was 27.5 years (7-87); left limb 155, right 117 cases; injury time ranged from 0.3 to 24 h (median 4 h). Conventional and modified OAR was applied on physical examination, subsequently radiography performed to determine the occurrence of fractures.The efficiency of the two methods were compared and analyzed. Results: Fractures were found in 100 cases (36.8%), 49 cases of ankle and 51 cases of foot fractures.With the imaging results as the standard, the sensitivity for conventional and modified OAR were 93.0% and 100%, specificity were 9.9% and 8.7%, the positive predictive value were 37.5% and 38.9%, the negative predictive value were 70.8% and 100%, the accuracy were 40.4% and 42.3%, missed diagnosis rate were 7% and 0% respectively.The sensitivity, positive likelihood ratio, positive predictive value, negative predictive value, accuracy, negative likelihood ratio and missed diagnosis ratio were better than in modified OAR compared with Conventional OAR, while the specificity was slightly lower compared to Conventional OAR.The Kappa value of modified OAR was 0.065 (P>0.05), which is better than conventional OAR.Conventional OAR can reduce 6.3% (17/272) X-ray and modified OAR decline 5.5% (15/272). Conclusion: Modified OAR significantly reduces the rate of missed diagnosis of foot fractures, but its specificity is poor. Ultrasound can be assisted to improve the specificity and reduce the number of unnecessary X-rays.

[Study on feasibility of enhanced recovery after surgery combined with mobile microendoscopic discectomy-transforaminal lumbar interbody fusion in the treatment of lumbar spondylolisthesis].

Objective: To explore the feasibility of enhanced recovery after surgery (ERAS) combined with mobile microendoscopic discectomy-transforaminal lumbar interbody fusion (MMED-TLIF) in the treatment of lumbar spondylolisthesis and its influence on postoperative rehabilitation. Methods: From October 1 2014 to July 1 2016 , a cohort of 52 patients with lumbar spondylolisthesis who received the program of ERAS-MMED-TLIF were retrospectively reviewed in Department of Minimally Invasive Spine Surgery, Tianjin Hospital.The primary outcomes include the operation time, intraoperative blood loss, length of hospital stay, VAS score (low back pain and leg pain) and Oswestry Disability Index (ODI) at different follow-up time and complication.The height of intervertebral space and fusion rate were also recorded as radiographic indicators. Results: All cases had an average follow-up of 12 months. The mean operative time was (115±30) min with a mean blood loss of (100±35) ml.Compared with preoperative condition, VAS score of low back pain (6.3±3.3 vs 3.5±2.3, P<0.05), VAS score of leg pain (7.1 ± 4.2 vs 3.1 ± 2.6, P<0.05) and the ODI disability index score (43.5±9.6 vs 20.9±7.3, P<0.05) at the postoperative 24 h were decreased and the difference was statistically significant.The mean hospitalized time were (4.9±1.3) days with mean postoperative hospital stay (2.1±1.2) days.Fusion rate was 92.31% (48/52) at the last follow-up time. Conclusion: ERAS combined with MMED-TLIF is feasible in the treatment of lumbar spondylolisthesis, which can significantly reduce intraoperative bleeding, shorten the total length of stay and postoperative hospital stay, improve postoperative pain and promote rapid rehabilitation of patients after operation without increasing the operation time and influencing the long-term effect, it can be applied in clinical practice.

[Clinical application of 125I radioactive seeds brachytherapy in the treatment of the pediatric soft tissue sarcoma in head and neck].

Objective: To investigate the clinical application and preliminary results of 125I radioactive seeds brachytherapy in the comprehensive treatment of the pediatric soft tissue sarcoma in head and neck. Methods: A total of 24 pediatric patients with soft tissue sarcoma in head and neck were treated at Peking University School of Stomatology from April 2012 to July 2015. The data was collected and analyzed through statistical methods, which included the pathological type, gender, age, tumor location, volume, treatment and the clinical results after the application of 125I radioactive seeds brachytherapy. Results: Among the 24 patients, there were 18 patients of rhabdomyosarcoma, 2 Ewing's sarcoma, 2 fibrosarcoma, 1 synovial sarcoma and 1 malignant fibrous histiocytoma. After a follow-up of 9-48 months, 1 case of temporal rhabdomyosarcoma was observed to have a progression to the the lateral cranial base at the time of 12 months, 2 cases realized local control and systemic progression, the rest were completely relieved, and there was no recurrence during the follow-up period.The Kaplan-Meier survival analysis showed that the 1-year and 3-year local control rate were both 94.1%, the 1-year and 3-year event-free survival rate were 87.4% and 77.7%. Conclusion: The preliminary results indicate that 125I radioactive seeds brachytherapy play a very important role in the improvement of the local control rate and survival quality of the pediatric soft tissue sarcoma patient in head and neck, and it's a minimally invasive treatment with the advantage of accuracy andsmall side effects.

[Study on apoptosis mechanism of H9c2 cardiomyocytes induced by N, N-dimethylformamide].

Objective: To observe the change levels of nuclear factor-kappa B (NF-κB) p65 protein in cytoplasm and nuclear, phosphorylation of inhibitor of kappa B (p-IκB) protein and cytochrome C (Cyt-c) , cleaved cysteinyl aspartate specific proteinase-3 (Cleaved caspase-3) , B-cell lymphoma/leukemia-2 (Bcl-2) in cytoplasm in the process of N, N-dimethylformamide (DMF) -induced apoptosis in H9c2 cardiomyocytes, and explore the tentative mechanism of apoptosis. Methods: H9c2 cardiomyocytes were exposed to 200 mmol/L DMF. Western blotting was used to detect the protein expression levels of p65 in cytoplasm and nuclear, p-IκB after exposure for 0, 2, 4, 6, 8, 12 h, and the protein expression levels of Cyt-c, Cleaved caspase-3, Bcl-2 in cytoplasm after exposure for 0, 2, 4, 8, 12, 24 h. Immunofluorescencecytochemistry (IFC) was used to observe the location of Cyt-c after 200 mmol/L DMF exposure for different times. Results: The levels of p65 in cytoplasm and nuclear and p-IκB among groups were statistically significant (F were 7.79, 33.11, 90.25, respectively, all P<0.01) . Compared with the control group, the levels of p65 in cytoplasm of 2, 4, 6 h group were significantly decreased (all P<0.01) ; the levels of p65 in nuclear of 2, 4, 6, 8 h were significantly increased (all P<0.01) ; the levels of p-IκB of 2, 4, 6 h group were significantly increased (all P<0.01) . The levels of Cyt-c, Cleaved caspase-3 and Bcl-2 among groups were statistically significant (F were 51.42, 503.68, 73.37, respectively, all P<0.01) . Compared with the control group, the levels of Cyt-c of 8, 12 h group were significantly increased (both P<0.01) ; the levels of Cleaved caspase-3 of 2, 4, 8, 12, 24 h were significantly increased (all P<0.01) ; the levels of Bcl-2 of 2, 4, 8, 12, 24 h group were significantly decreased (all P<0.01) . IFC showed that Cyt-c was released from the mitochondria to the cytoplasm gradually as the extension of the exposure time. Conclusion: NF-κB signaling pathway and mitochondrial pathway are involved in the mechanism of DMF-induced apoptosis in H9c2 cardiomyocytes.

[Complications of open reduction and internal fixation versus external fixation for unstable distal radius fractures: a meta-analysis].

Objective: To make a systematic assessment of the complications of open reduction and internal fixation versus external fixation for unstable distal radius fractures. Method: A computer-based online search of PubMed, ScienceDirect, EMBASE, BIOSIS, Springer and Cochrane Library were performed.The randomized and controlled trials of open reduction and internal fixation versus external fixation for unstable distal radius fractures were collected.The included trials were screened out strictly based on the criterion of inclusion and exclusion.The quality of included trials was evaluated.RevMan 5.0 was used for data analysis. Result: A total of 17 studies involving 1 402 patients were included.There were 687 patients with open reduction and internal fixation and 715 with external fixation.The results of Meta-analysis indicated that there were statistically significant differences with regard to the postoperatively total complications, infection, malunion, tendon rupture (I2=8%, RR=0.77(95%CI 0.65-0.91, Z=3.10, P<0.05). There were no statistically significant differences observed between two approaches with respect to nounion, re-operation, complex regional pain syndrome, carpal tunnel syndrome, neurapraxia, tendonitis, painful hardware, scar(P>0.05). Conclusion: Postoperative complications are present in both open reduction and internal fixation and external fixation.Compared with external fixation, open reduction and internal fixation is lower in total complications postoperatively, infection and malunion, but external fixation has lower tendon rupture incidence.