OBJECTIVE To investigate the relationship of long non-coding RNA （LncRNA） metastasis-associated lung adenocarcinoma transcript 1 （MALAT1） and doxorubicin （DOX） resistance in osteosarcoma （OS） cells. METHODS MG-63 and MG-63/DOX cells were treated with different concentrations of DOX （0， 0.01， 0.05， 0.1， 1 μmol/L）， and survival rates and half maximal inhibitory concentration were determined using CCK-8 assay. The expressions of LncRNA MALAT1 in MG-63 and MG-63/ DOX cells were detected by real-time quantitative fluorescence PCR. MG-63/DOX cells were divided into Control group， knocking down LncRNA MALAT1 negative control （sh-NC） group， sh-MALAT1 group， sh-MALAT1+anti-NC group， and sh-MALAT1+ anti-miR-154-5p group. The expressions of LncRNA MALAT1， miR-154-5p and cyclin D1 （CCND1） mRNA in MG-63/DOX cells of each group were detected. The effects of knocking down LncRNA MALAT1 on the proliferation， migration， invasion， and apoptosis of MG-63/DOX cells were detected by CCK-8 assay， scratch test， Transwell experiment and flow cytometry， respectively. The expression of proliferating cell nuclear protein （PCNA） and CCND1 protein in MG-63/DOX cells was detected by Western blot assay. Interactions between LncRNA MALAT1 and miR-154-5p， miR-154-5p and CCND1 were detected by dual luciferase reporter gene experiment. RESULTS Compared with 0 μmol/L DOX， 0.01， 0.05， 0.1 and 1 μmol/L DOX could reduce the survival rates of MG-63 and MG-63/DOX cells （except for 0.01 μmol/L DOX） （P＜0.05）， IC50 were 0.07 and 0.13 μmol/L， respectively. The survival rate， cell migration number and invasion number of MG-63/DOX cells， scratch closure rate， mRNA expressions of LncRNA MALAT1， mRNA and protein expressions of CCND1， and PCNA protein expression in sh-MALAT1 group were significantly lower than sh-NC group and Control group； the apoptosis rate and miR-154-5p expression were significantly higher than sh-NC group and Control group （P＜0.05）. sh-MALAT1+anti-miR-154-5p group was able to reverse the aforementioned biological effects in sh-MALAT1 group （P＜0.05）. In MG-63/DOX cells transfected with both MALAT1-wild type （WT） and CCND1-WT， the luciferase activity in the miR-154-5p mimic group was significantly lower than mimic negative control group （P＜ 0.05）. CONCLUSIONS Knocking down LncRNA MALAT1 can inhibit the DOX resistance of OS cells， and its mechanism may be targeting the miR-154-5p/CCND1 axis.

Objective:To explore the related risk factors for systemic embolism (SE) in patients aged≥75 years with non-valvular atrial fibrillation (NVAF).Methods:A case-control study. NVAF patients aged≥75 years who were hospitalized at the First Affiliated Hospital of Xinjiang Medical University from October 2018 to October 2020 were divided into no SE ( n=1 127) and SE ( n=433) groups according to the occurrence of SE after NVAF. Multivariate logistic regression was used to analyze SE-related factors in patients with NVAF without anticoagulation treatment. Results:In the multivariate model, the following factors were associated with an increased risk of SE in patients with NVAF: history of AF≥5 years [odds ratio ( OR)=2.75, 95% confidence interval ( CI) 1.98-3.82, P<0.01], lipoprotein(a)>300 g/L ( OR=2.07, 95% CI 1.50-2.84, P<0.01), apolipoprotein (Apo)B>1.2 g/L ( OR=1.91, 95% CI 1.25-2.93, P=0.003), left ventricular ejection fraction (LVEF) of 30%-49% ( OR=2.45, 95% CI 1.63-3.69, P<0.01), left atrial diameter>40 mm ( OR=1.54, 95% CI 1.16-2.07, P=0.003), and CHA 2DS 2-VASc score≥3 ( OR=15.14, 95% CI 2.05-112.13, P=0.01). ApoAI>1.6 g/L was negatively correlated with the occurrence of SE ( OR=0.28, 95% CI 0.15-0.51, P<0.01). Conclusions:History of AF≥5 years, lipoprotein(a)>300 g/L, elevated ApoB, left atrial diameter>40 mm, LVEF of 30%-49%, and CHA 2DS 2-VASC score≥3 are independent risk factors for SE whereas ApoAI>1.6 g/L is a protective factor against SE in patients with NVAF.

Objective:To improve the understanding of autoimmune lymphoproliferative syndrome (ALPS).Methods:The clinical data of the proband and his family members in Children's Hospital Affiliated to Zhengzhou University in August 2018 were retrospectively analyzed, and the peripheral blood DNA of the proband, his parents and siblings was extracted. High-throughput next-generation sequencing was used to make gene analysis and validation. Phenotype and genotype of them were also analyzed. Relevant literature was reviewed.Results:The proband was a 1-year and 1-month old boy with hemolytic anemia, thrombocytopenia and splenomegaly as the main manifestations. The double negative T cells and the Vitamin B 12 of the proband were significantly increased and the autoantibodies were positive. The boy's father had a history of splenomegaly. His elder brother and sister had similar clinical manifestations. The results of next-generation sequencing showed that the FAS gene frameshift mutation (c.648delT) was detected in this boy and his father, elder brother and sister, which was a new mutation. After immunosuppressive treatment, the symptoms of the boy improved and the blood cells increased. Conclusions:The frameshift mutation of FAS gene may be the cause of the disease in this ALPS pedigree. Clinically, it is necessary to consider ALPS for children with unexplained hemocytopenia and hepatosplenomegaly. Double-negative T cells, autoantibodies, Vitamin B 12 should be tested, and high-throughput gene sequencing should be performed if necessary.

Objective:To investigate the effect of miRNA-1-3p (miR-1-3p) on expression of myocyte enhancer factor 2A (MEF2A) and the biological function of osteosarcoma cells.Methods:The tumor tissues and adjacent normal tissues of 20 patients with osteosarcoma who were clinically diagnosed in Shanxi Provincial Cancer Hospital from January 2019 to January 2020 were collected, and the expression of miR-1-3p in the samples was detected by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). The expression of miR-1-3p in osteosarcoma cell lines U2-OS, SAOS-2, MG63, SW1353 and human normal osteoblast cell line hFOB1.19 was detected by qRT-PCR, then the cell line with the lowest expression of miR-1-3p was selected for follow-up experiments. An overexpression miR-1-3p vector was constructed (miR-1-3p mimcs). The miR-1-3p overexpression group was transfected with miR-1-3p mimcs, and the control group was transfected with empty vector (miR-1-3p nc). CCK-8 method was used to detect the proliferation activity of cells; flow cytometry was used to detect the changes of cell apoptosis and cell cycle. miRwalk database was used to predict the miR-1-3p target gene, and the target gene was verified by dual-luciferase reporter gene assay; Western blot was used to detect the expression of MEF2A protein in cells of each group.Results:Compared with adjacent tissues, the expression of miR-1-3p in osteosarcoma tissues was down-regulated (0.31±0.14 vs. 0.62±0.21), and the difference was statistically significant ( t = 5.31, P<0.01). The expression of miR-1-3p in U2-OS cells was the lowest; compared with the control group, the proliferation activity of U2-OS cells was inhibited in miR-1-3p overexpression group (48 h absorbance value 0.56±0.01 vs. 0.77±0.03, t = 2.77, P<0.01; 72 h absorbance value 0.87±0.02 vs. 1.40±0.03, t = 2.93, P<0.01); G 1/S cell cycle arrest increased [G 1 phase (38.24±0.55)% vs. (32.11±0.80)%, t = 9.27, P = 0.01; S phase (61.24±0.90)% vs. (67.78±0.83)%, t = 7.52, P = 0.02]; early apoptotic rate increased [(11.20±0.12)% vs. (1.50±0.12)%, t = 2.91, P<0.05], miRwalk database predicted that the miR-1-3p target gene was MEF2A. The result of dual-luciferase reporter gene assay showed that miR-1-3p bound to MEF2A 3'UTR, and the luciferase activity of U2-OS cells in miR-1-3p overexpression group was lower than that in the control group (renilla luciferase/firefly luciferase activity ratio 0.53±0.06 vs. 1.00±0.04, t = 4.04, P < 0.05). Western blot showed that the expression of MEF2A protein in U2-OS cells of miR-1-3p overexpression group was lower than that of the control group (protein relative expression 0.41±0.14 vs. 0.77±0.12, t = 3.93, P < 0.05). Conclusions:The low expression of miR-1-3p may be associated with the proliferation, apoptosis and cycle changes of osteosarcoma cells. miR-1-3p can negatively regulate the expression of MEF2A protein and regulate the occurrence and development of osteosarcoma.

Objective:To explore the application value of heparin-binding protein (HBP) in the early diagnosis and assessment of severe adenovirus pneumonia.Methods:A total of 90 children diagnosed with adenovirus pneumonia admitted in the Department 1 of Emergency and Pediatric Intensive Care Unit 1 in Hunan Children′s Hospital from January 2019 to March 2020 were recruited.HBP levels in children with adenovirus pneumonia were detected.The correlation between HBP with white blood cell count (WBC), neutrophil ratio (N), C-reactive protein (CRP), interleukin-6(IL-6) and erythrocyte sedimentation rate (ESR) were examined.Receiver operating characteristic(ROC) curve analysis was conducted to explore the value of HBP in the early diagnosis and assessment of severe adenovirus pneumonia.Children with adenovirus pneumonia were divided into severe adenovirus pneumonia group (severe group) and non-severe adenovirus pneumonia group (non-severe group) according to their severity.Those in the severe group were further divided into bronchiolitis obliterans(BO) group and non-BO group according to the occurrence of BO.Results:(1) The HBP level in children with adenovirus pneumonia was (49.47±34.19) μg/L, which was significantly higher in the severe group than that of non-severe group[(82.88±44.02) μg/L vs.(35.15±13.08) μg/L, t=15.349, P<0.05]. Children in the severe group were significantly younger, and they had a significantly longer length of stay, lower Pediatric Critical Illness Scores (PCIS), and higher inflammatory markers like HBP, WBC, N, CRP, IL-6, and ESR compared with those of the non-severe group (all P<0.05). No significant difference in the procalcitonin (PCT) level was detected between groups.(2) The HBP was positively correlated with inflammatory markers like WBC ( r=0.38, P<0.05), N ( r=0.26, P<0.05), CRP ( r=0.47, P<0.05), IL-6 ( r=0.76, P<0.05), and ESR ( r=0.35, P<0.05). However, HBP did not have a significant correlation with PCT ( r=0.097, P>0.05). (3) In the severe group, the HBP level of the children with invasive mechanical ventilation, oxygenation index(P/F index)≤ 200 mmHg (1 mmHg=0.133 kPa) and BO was significantly higher than that of the non-invasive mechanical ventilation, P/F index> 200 mmHg and non-BO (all P<0.05). (4) The area under the ROC curve of HBP, WBC, N, CRP, ESR and IL-6 in predicting the severity of adenovirus pneumonia were 0.915, 0.748, 0.770, 0.740, 0.820 and 0.798, respectively.When the cut-off value of HBP was 45 μg/L, the sensitivity and specificity of HBP were 81.48% and 85.71%, respectively. Conclusions:As an inflammatory mediator, HBP is involved in the inflammatory response of the body.It may be a useful new marker for the early diagnosis of severe adenovirus infection, which also has a certain value in the evaluation of the severity and prognosis of the disease.The findings provide a basis for early clinical intervention and treatment of adenovirus infection in children.

Objective:To analyze the clinical phenotypes and molecular genetic characteristics of hereditary spherocytosis (HS) in children.Methods:The clinical data of children with HS in the Affiliated Children′s Hospital of Zhengzhou University from June 2014 to June 2018 were collected and analyzed retrospectively.The peripheral blood samples of 20 children and their parents were collected and targeted capture or whole exon group sequencing was performed using the second generation sequencing technique, and the suspicious mutation sites were verified by Sanger sequencing.Results:A total of 43 pediatric patients (23 males, 20 female) were included in this study.The median age of onset of 1 year old and 11 months (1 month to 10 years old). Pallor (27/43 cases, 62.79%) was the main symptom.Typical clinical manifestations were anemia (36/43 cases, 83.72%), jaundice (35/43 cases, 81.40%), splenomegaly (33/43 cases, 76.74%), and hepatomegaly (27/43 cases, 62.79%). The proportion of spherical erythrocytes in peripheral blood of 23 cases (23/43 cases, 53.49%)≥ 0.1, and the proportion of spherical erythrocytes in bone marrow smears of 17 cases (39.53%) was ≥ 0.1, and 20 cases were positive in erythrocyte osmotic fragility test (20/43 cases, 46.51%). Comparison of 29 children before and after blood transfusion: post-transfusion hemoglobin (Hb) [(88.69±11.22) g/L vs. (78.24±14.47) g/L], mean corpuscular volume (MCV) [(89.37±7.15) fL vs.(84.08±7.49) fL], mean corpuscular hemoglobin (MCH) [(29.12±2.70) pg vs.(27.36±1.95) pg], mean corpuscular hemoglobin corpuscular (MCHC) [(361.79±32.27) g/L vs.(356.31±31.43 ) g/L] were increased, and the total bilirubin (TB) level was lower after blood transfusion than before blood transfusion [(33.27±16.42) μmol/L vs.(41.58±15.40) μmol/L], the differences were statistically significant ( t=-3.538, -5.187, -5.412, -7.404, 2.527, all P<0.05). Seven children underwent splenectomy: postoperative Hb[(116.00±5.54) g/L vs.(75.71±9.96) g/L], MCH[(29.87±1.62) pg vs.(24.61±1.65) pg], MCHC [(391.14±12.99) g/L vs.(315.14±51.99) g/L] all increased, the difference was statistically significant ( t=-9.234, -4.330, -4.031, all P<0.05). The Hb in the operation group was significantly higher than that in the blood transfusion group, including the treatment effect of operation was more significant ( t=-9.247, P<0.05). Thirteen pathogenic mutations (11 new mutations not reported) were detected in 3 genes ( ANK1, SPTB and SPTA1) in 13 children, 8 of who were mutations in ANK1, 4 cases of which were mutations in SPTB and 1 case of which was mutation in SPTA1.Among the 13 mutations, 12 cases were de novo except one inherited from the mother. Conclusions:Anemia and pallor are the initial symptoms of HS in children.Blood transfusion can temporarily improve the symptoms of anemia and jaundice.Splenectomy is an effective and fundamental treatment.The gene diagnosis of 13 pediatric patients got gene diagnosis, which provided basis for genetic consultation of the next fetus.Eleven new mutations were found in ANK1, SPTB, SPTA1 genes, which exten-ded the gene mutation spectrum of HS.

Objective To explore the predicted performance of cTnI for outcome or severity in children with sepsis.Methods 374cases of children with sepsis were collected in pediatric intensive care unit (PICU) in our hospital from August 2012to June 2015.The patients were dividided into the common sepsis group, severe sepsis group and sepsis shock group according to the sepsis severity, and improved group, uncured group and death group according to outcome, and the cTnI>0.01μg/mL group and the cTnI≤0.01μg/mL group according to the levels of cTnI.Data on cTnI, PCT, CRP, Cr, Lac, PaO2/FiO2, BUN, PT, INR, WBC and PLT were collected in this study.Results The level of cTnI was significantly higher in children with septic shock (P<0.05) .The level of cTnI in improved group was significantly lower than those of uncured group and death group (P<0.05) .The incidence of severe sepsis and septic shock in the cTnI>0.01μg/mL group was significantly significantly higher than that of the cTnI≤0.01μg/mL group.The levels of Lac, PT and INR in the cTnI>0.01μg/mL group were significantly higher than that of the cTnI≤0.01μg/mL group (P<0.05) .A positive correlation between the level of cTnI and Lac (r=0.324) , or PT (r=0.291) , or INR (r=0.340) were found in the study (P<0.05) .Conclusion Sepsis is prone to be associated with myocardial injury, which is related to the severity and prognosis of sepsis.Insufficient circulatory perfusion, metabolic imbalance and abnormal coagulation function may be the reasons for the rise of cTnI and myocardial injury in children with sepsis.

Objective To study the correlation between serum miRNA-21 (miR-21) level and chemosensitivity and prognosis of osteosarcoma patients. Methods A total of 68 osteosarcoma patients who were treated in Shanxi Provincial Cancer Hospital from August 2016 to August 2017 were selected. All patients received routine chemotherapy after admission. They were followed up for one year. According to the effectiveness of chemotherapy, they were divided into effective group (52 cases) and ineffective group (16 cases). The general clinicopathological characteristics of the two groups were recorded. Univariate logistic regression model was used to analyze the serum miR-21 level and chemosensitivity in osteosarcoma patients. The prognostic efficacy of serum miR-21 was evaluated by receiver operating characteristic (ROC) curve. Results The serum miR-21 expression in the effective group was higher than that in the ineffective group (6.6 ±0.7 vs. 5.2 ±0.7), and the tumor metastasis rate in the effective group was lower than that in the ineffective group [33.9％ (18/52) vs. 37.5％ (6/16)], and the differences between the two groups were statistically significant (both P< 0.05). There were no significant differences in gender, age, body mass index (BMI), tumor location, size of tumors and TNM stage between the two groups (all P>0.05). Univariate logistic regression model showed that serum miR-21 and metastasis were the factors affecting chemosensitivity (both P<0.05). ROC curve showed that the area under the curve (AUC) of using serum miR-21 to predict the prognosis of osteosarcoma patients was 0.774. Youden index indicated that the best cut-off points of serum miR-21 and chemosensitivity for predicting the prognosis of osteosarcoma patients was 6.25. Conclusion Serum miR-21 level in osteosarcoma patients is significantly correlated with chemosensitivity and prognosis, and it can be used as an effective index to predict the effect of chemotherapy and prognosis of patients with osteosarcoma.

Objective To analyze the feasibility and safety in application of resectoscope combined with laparoscopy in the operation treatment of paraganglioma of urinary bladder.Methods 7 cases patients with paraganglioma of urinary bladder treated in our hospital from November 2014 to August 2018 were analyzed retrospectively.There were 5 males and 2 females,average age of 31.1 years (22-37 years),average body mass index was 22.3 kg/m2 (18.3-22.5 kg/m2).All the 7 cases patients complained of dizziness and palpitation after urination,average basal systolic blood pressure was 111.8 mmHg (97-124 mmHg),the average fluctuation of systolic blood pressure before and after urination was 64.9 mmHg(28-91 mmHg),the CT and cystoscopy prompt bladder tumor,the average diameter was 2.7 cm(2.1-3.5 cm).The average of plasma norepinephrine was 706.3 pg/ml(330-997 pg/ml);the average of plasma dopamine was 101.1 pg/ml(44-145 pg/ml);the average of 24h urinary vanilmandelic acid was 13.4 mg/24h (10.3-16.1 mg/24h).All the patients has controlled the blood pressure and dilate the blood vessels with phenoxybenzamine hydrochloride,accepted the operation of resectoscope combined with laparoscopy partial cystectomy and bladder sutura per abdomen after ample dilatancy.The patients had lithotomy position with trendelenburg,preparation of gas peritoneal cavity by transabdominal,inside the resectoscope by transurethral at the same time,mutilated bladder mucosa beside 1cm at the edge of the tumor,and cut full thickness bladder wall,take the extraperitional fat as the standard procedure;we could see the cutting edge clear at this time by laparoscopy,cut off the pelvic peritoneum,extraperitional fat and the tumor.The sample placed in bladder,close the bladder with absorbable or barbed wires,take out the sample by resectoscope.Results All the 7 cases patients operation was successfully completed,no cases has been transfered to open.The average time of operation is 85.3 min(65-100 min),the average amount of bleeding is 27.9 ml(10-50 ml).The average fluctuation of systolic blood pressure is 8.7 mmHg(6-15 mmHg).Bladder washout was stopped 24h after operation,catheter was removed 1 weeks after operation.There is no obvious complications occurred.The average hospital stay is 3.7 days (3-5 days).The average pain score of 4 cases 4h after operation is 3.8 (2-5),reevaluation 24h after operation is 2.3 (1-4).The average follow-up time is 7.9 months(2-15 months).All the 7 cases patients clinical symptoms disappeared,there is no fluctuation of systolic blood pressure before and after urination,there is no recurrence of the tumor.Conclusions To the paraganglioma of urinary bladder in fundus of bladder or anterior wall of bladder,we can accurate resection tumor by resectoscope combined with laparoscopy,reduce blood pressure fluctuations,reduce the surgical trauma and the distress of patients.It is a safety and effective minimally invasive surgery.

Objective To evaluate GGT in combination with B ultrasound for the diagnosis of biliary atresia (BA) infants suffering from obstructive jaundice.Methods A retrospective analysis was made on 69 sick infants including 55 BAs and 14 non-BAs as identified by intraoperative cholangiography.The preoperative laboratory GGT and ultrasound data were collected and analyzed.The sensitivity,specificity,positive predictive value,negative predictive value and accuracy were compared.Results BA patients had significantly higher GGT than Non-BA patients (t =-4.164,P ＜ 0.05).The sensitivity,specificity,positive predictive value,negative predictive value and accuracy of GGT ＞ 306 U/L were 69.1％,92.9％,97.4％,43.3％,73.9％,respectively.In BA group,abnormal gallbladder was significantly associated with proadening portal vein,broadening hepatic artery compared with Non-BA patients (x2 =9.995,P ＜0.05).The accuracy of abnormal gallbladder on ultrasound was 78.3％.When two method combined for the diagnosis of BA,the sensitivity,specificity,positive predictive value,negative predictive value were 92.7％,92.9％,98.1％ and 76.5％ and accuracy can reach 92.8％.Conclusions For obstructive jaundice infants with GGT ＞ 306 U/L and abdominal gallbladder ultrasound finding intraoperative cholangiography should be carried out to make definite diagnosis of BA.