Objective:To analyze thalassemia genotypes and distribution of children in Wuzhou Guangxi,and evaluate the diagnostic value of HbA2 in children's thalassemia screening,so as to provide scientific evidence for the prevention and control strategies of thalassemia.Methods:Four hundred and fifty-eight children suspected with thalassemia in Wuzhou were enrolled from March 2017 to June 2022.The level of HbA2 was detected using Bio-Rad VARIANT Ⅱ Hb analysis system.The deletion of α-thalassemia was measured with gap-PCR assay,and the point mutation of α-and β-thalassemia was tested with DNA reverse dot blot hybridization assay.The receiver operating characteristic(ROC)curve was used to analyze the diagnostic value of HbA2 for children's thalassemia.Results:A total of 304 thalassemia carriers were detected in 458 children,accounting for 66.38％.One hundred and seventy-five cases were defined to be α-thalassemia,with the main type of--SEA/αα(54.86％).Thirty-six cases were defined to be intermediate α-thalassemia,with the main type of-α3.7/--SEA(9.72％).In 108 cases with β-thalassemia,βCD41-42/βN was the main type,accounting for 49.07％,followed by βIVS-Ⅱ-654/βN(14.81％).Seven cases were moderate/severe β-thalassemia(predominantly β-28/β-28 and βCD41-42/βCD17)Twenty-one genotypes of α-and β-thalassemia were found in the children.There was significant difference of HbA2 level between the children with different types of thalassemia and healthy controls(all P＜0.001).ROC curve analysis showed that the sensitivities of HbA2 for α-thalassemia,β-thalassemia and αβ-thalassemia were 74.3％,82.4％and 85.7％,with the optimal cut-off values of 2.60％,3.60％and 3.70％,respectively,the specificities were 64.3％,96.1％and 96.8％,and the area under the curve were 0.690,0.887 and 0.916,respectively.Conclusion:The thalassemia genotypes of children in Wuzhou are diverse.It is necessary to further strengthen the prevention and control measure of thalassemia to reduce birth defects and improve birth quality.

OBJECTIVES: To investigate the risk factors for acute kidney injury (AKI) after hematopoietic stem cell transplantation (HSCT) in children. METHODS: A retrospective analysis was performed on the medical data of 111 children who underwent HSCT from January 2018 to January 2020. A multivariate logistic regression analysis was used to identify the risk factors for AKI. The Kaplan-Meier survival analysis was used to compare the prognosis in children with different grades of AKI. RESULTS: Graft-versus-host disease (grade Ⅱ-Ⅳ) (OR=4.406, 95%CI: 1.501-12.933, P=0.007), hepatic veno-occlusive disease (OR=4.190, 95%CI: 1.191-14.740, P=0.026), and thrombotic microangiopathy (OR=10.441, 95%CI: 1.148-94.995, P=0.037) were closely associated with the development of AKI after HSCT. The children with stage Ⅲ AKI had a lower 1-year survival rate than those without AKI or with stage Ⅰ AKI or stage Ⅱ AKI (28.6%±12.1% vs 82.8%±5.2%/81.7%±7.4%/68.8%±11.6%; P<0.05). CONCLUSIONS Children with stage Ⅲ AKI after HSCT have a higher mortality rate. Graft-versus-host disease, hepatic veno-occlusive disease, and thrombotic microangiopathy are closely associated with the development of AKI after HSCT.
Child ; Humans ; Retrospective Studies ; Hematopoietic Stem Cell Transplantation/adverse effects* ; Graft vs Host Disease/complications* ; Risk Factors ; Acute Kidney Injury/therapy* ; Thrombotic Microangiopathies/complications*

ObjectiveTo investigate the influence of E2F transcription factor 2 （E2F2） on the cell adhesion of multiple myeloma cells by combining bioinformatics and cellular and molecular experiments. MethodsFirstly， clinical samples and GEO database were used to analyze the relationship between the E2F2 expression and the prognosis of myeloma patients. Then， differential genes were screened from the RNA-seq data of E2F2 knockdown myeloma cell line MM.1S. The GO biological function enrichment and KEGG signal pathway analysis were performed on these differential genes. Meanwhile， the protein interaction network was analyzed by using the string website. The E2F2 stable knockdown MM.1S cell line were constructed， and the expression level of E2F2 was measured by Western Blotting and qRT-PCR. The effect of stable knockdown E2F2 on the adhesion of myeloma cells was detected by cell adhesion experiment， and the expression of cell adhesion related genes were detected by qRT-PCR. The effect of stable E2F2 knockdown on myeloma cell migration was detected by cell migration assay. ResultsThe expression of E2F2 was significantly increased in clinical samples of myeloma patients， and the high expression of E2F2 is correlated with poor prognosis of myeloma patients in the GEO database. The analysis of RNA-seq data revealed 815 differentially expressed genes， of which 508 genes were up-regulated and 307 genes were down-regulated. These genes were closely related to the cell adhesion and angiogenesis. The construction of E2F2 knockdown MM.1S cell lines were verified by Western Blotting and qRT-PCR. Knockdown of E2F2 significantly increased the adhesion level of MM.1S cells and elevated the expression of FN1， PECAM1， ICOSLG and other cell adhesion related genes， while weakening the invasion ability of MM.1S cells， P < 0.05. ConclusionsThrough bioinformatics analysis of the RNA-seq data， we found that the transcription factor E2F2 is closely related to the cell adhesion in myeloma. Knockdown of E2F2 in the MM.1S cell line promotes the expression of cell adhesion related genes， increases the cell adhesion level， and inhibites cell migration ability.

In this study, we improved the culture method of mouse hippocampal primary microglia to obtain hippocampal ramified microglia with high activity and purity, which were resemble to the resting status of normal microglia in healthy brain in vivo. Hippocampal tissue was excised from 2-4-week-old SPF C57BL/6J mice and cut into pieces after PBS perfusion, and then manually dissociated into the single-cell suspension by using Miltenyi Biotec's Adult Brain Dissociation Kit. The tissue fragments such as myelin in the supernatant were removed by debris removal solution in the kit. The cell suspension was incubated with CD11b immunomagnetic beads for 15 min at 4 °C. To obtain high-purity microglia, we used two consecutive cell-sorting steps by magnetic activated cell sorting (MACS). After centrifugation, the cells were resuspended and seeded in a 24-well culture plate. The primary microglia were cultured with complete medium (CM) or TIC medium (a serum-free medium with TGF-β, IL-34 and cholesterol as the main nutritional components) for 4 days, and then were used for further experiments. The results showed that: (1) The cell viability was (56.03 ± 2.10)% by manual dissociation of hippocampus; (2) Compared with immunopanning, two-step MACS sorting allowed for efficient enrichment of microglia with higher purity of (86.20 ± 0.68)%; (3) After being incubated in TIC medium for 4 d, microglia exhibited branching, quiescent morphology; (4) The results from qRT-PCR assay showed that the levels of TNF-α, IL-1β and CCL2 mRNA in TIC cultured-microglia were similar to freshly isolated microglia, while those were much higher in CM cultured-microglia after incubation for 4 d and 7 d (P < 0.05). Taken together, compared to the conventional approaches, this modified protocol of mouse hippocampal primary microglia culture by using MACS and TIC medium enables the increased yield and purity of microglia in the quiescent state, which is similar to normal ramified microglia in healthy brain in vivo.
Animals ; Cell Culture Techniques ; methods ; Cell Separation ; methods ; Cells, Cultured ; Hippocampus ; Magnetics ; Mice ; Mice, Inbred C57BL ; Microglia ; cytology

OBJECTIVE: To compare the clinical pregnancy rates between two types of endometrial preparation protocolsnatural cycle（NC）and hormone replacement cycle（HRT）-in patients with thin endometrium in the frozen-thawed embryo transfer（FET）cycles.METHODS: From January 2012 to December 2018,FET patients with endometrial thickness ≤7 mm on the day of human chorionic gonadotropin（h CG）trigger in Reproductive Medicine Center of the First Affiliated Hospital of Sun Yat-sen University were selected as research subjects.According to the endometrial preparation protocols,they were divided into NC group and HRT group.Totally 117 pairs were successfully matched using the propensity score matching method.The matching variables were age,embryo type and number of transferred embryos,and the embryo implantation rate and clinical pregnancy rate of the two matched groups were compared.RESULTS: There was no significant difference in embryo implantation rate（36.47% vs. 39.03%）or clinical pregnancy rate（44.40% vs. 52.10%）between NC group and HRT group（P> 0.05）.CONCLUSION: NC group and HRT group had similar pregnancy rate in patients with thin endometrium in FET cycles.Individualized protocols can be adopted according to the characteristics of patients with thin endometrium.

Objective: To evaluate the clinical characteristics and survival outcomes of patients with de novo grade 3 or transformed follicular lymphoma (FL). Methods: Fifty-two patients treated at Peking University Cancer Hospital between January 2009 and September 2017 were assessed, including 28 patients with FL 3A grade, 13 patients with FL 3B grade, 11 patients with transformed FL. Baseline characteristics, survival and prognostic factors were analyzed. Results: ① Twenty-six male and 26 female patients were enrolled, including 28 patients with FL 3A grade, 13 patients with FL 3B grade, 11 patients with transformed FL. ②The 3-year progression-free survival (PFS) and overall survival (OS) for the entire cohort were 56.0% and 80.6%, respectively. Patients with international prognostic index (IPI) score 0-1 demonstrated significantly better 3-year PFS (80.3% vs 20.1%; t=18.902, P<0.001) and OS (95.7% vs 57.0%; t=10.406, P<0.001) than patients with IPI score 2-3. Three-year PFS (94.1% vs 37.2% vs 25.2%; P=0.002) and OS (100.0% vs 76.0% vs 59.8%; P=0.020) were also significantly different among patients with FLIPI 1 score 0-1, 2, ≥3. FLIPI 2 score was also identified as a prognostic factor for 3-year PFS (68.4%, 0, 0; P=0.001) and OS(87.5%, 76.2%, 0; P=0.003). ③Multivariate analysis indicated a significant association of PFS (HR=3.536, P=0.015) and OS (HR=15.713, P=0.015) with IPI. FLIPI 2 was associated with OS (score 0-1, HR=0.078, P=0.007; score 2, HR=0.080, P=0.022). Conclusion: De novo grade 3 or transformed FL might be a group of curable disease with current treatment strategies. IPI is still a prognostic tool in this scenario.
Disease-Free Survival ; Female ; Humans ; Lymphoma, Follicular ; Male ; Neoplasm Recurrence, Local ; Prognosis ; Retrospective Studies ; Survival Analysis

BACKGROUND:At present,most of the experiments on the treatment of skin wound by mesenchymal stem cells have been performed in rats,mice and rabbits,while the research on skin wound treatment by canine bone marrow mesenchymal stem calls is less reported.OBJECTIVE:To observe the effect of canine bone marrow mesenchymal stem calls on skin wound healing.METHODS:A 3 cmx3 cm wound was made on the both sides of the scapula and buttocks of the dog,with the right side as experimental group and the left side as control group.After the wound was made,allogeneic canine bone marrow mesenchymal stem calls suspension was injected subcutaneously around the wound in the experimental group on the 1st and 3rd days.The control group was injected subcutaneously around the wound with mesenchymal stem call culture medium on the 1st and 3rd days after the wound was made.Wound healing was observed dynamically in both groups.RESULTS AND CONCLUSION:In the 1st week,there were pale yellow inflammatory substances in the wound of two groups indicating obvious inflammations.Compared with the control group,the inflammatory substances were fewer and the growth rate of the granulation tissue was faster in the experimental group.From the 2nd week until the wound healing,epithelialization on the wound became obvious following the formation of the granulation tissue,which was mainly displayed by the formation of fresh epithelial tissues from the surrounding to the wound center.The epithelialization time of the experimental group was earlier than that of the control group,and the wound area of the experimental group was smaller than that of the control group.In the 3rd week,the wound in the experimental group healed completely,and became smoother than that in the control group.The wound area of the experimental group was slightly smaller than that of the control group on the 8th and 12th days after cell transplantation,and the healing speed of the experimental group was slightly faster than that of the control group,but there was no significant difference between the two groups.Our findings indicate that the transplantation of canine bone marrow mesenchymal stem cells has the possibility or trend to promote skin wound healing.

OBJECTIVETo explore possible interrelationships among resistance to peer pressure, risky decision-making and health risk behaviors among young adolescents.

METHODSBased on the cluster sampling method, the participants who were recruited from 5 junior middle schools in Guangzhou and 3 junior middle schools in Shenyang city on October, 2010, were administered to complete the questionnaire concerned with their experiences with drinking and smoking during the past 30 days preceding the survey, and the hours using computer daily both in weekdays and in weekend. The level of resistance to peer influence and risky decision-making were assessed by Resistance to peer influence scale (RPIS) and Youth decision-making questionnaire (YDMQ). Logistic regression was used to explore possible interrelationships among resistance to peer influence, risky decision-making and health risk behaviors among young adolescents.

RESULTSA total of 1985 questionnaires were valid, including 1001(50.4%) boys and 984 (49.6%) girls. About 27.1% (537/1985) junior middle school students reported having health risk behaviors, boys' (30.7%, 307/1001) was higher than girls' (23.4%, 230/984) with significant gender difference (P < 0.05). The prevalence of smoking, drinking during the past 30 days before the survey and using computer over 3 hours daily in weekdays and in weekend were 5.1% (102/1985), 14.3% (284/1985), 3.5% (70/1985) and 13.7% (272/1985), respectively. The rate of drinking, using computer over 3 hours daily in weekdays and in weekend were higher in males (16.4% (164/1001), 4.5% (45/1001), 16.2% (162/1001)) than those in females (12.2% (120/984), 2.5% (25/984), 11.2% (110/984)) (P < 0.05). The scores of RPIS and YDMQ of the two cities adolescents were 2.82 ± 0.39 and 1.68 ± 0.62. The students reported smoking, drinking during the past 30 days before the survey and using computer over 3 hours daily in weekend gained lower RPIS scores (2.43 ± 0.40, 2.61 ± 0.41, 2.77 ± 0.40) than their counterparts who didn't report these kind of health risk behaviors (2.84 ± 0.38, 2.85 ± 0.38, 2.82 ± 0.39)(P < 0.05). And those reported smoking, drinking during the past 30 days before the survey and using computer over 3 hours daily in weekdays and in weekend gained higher YDMQ scores (2.38 ± 0.66, 2.06 ± 0.66, 1.97 ± 0.72, 1.84 ± 0.64, respectively) than their counterparts who didn't report these kind of health risk behaviors (1.64 ± 0.38, 1.61 ± 0.58, 1.67 ± 0.61, 1.65 ± 0.61, respectively) (P < 0.05). After adjusting gender, area, parental education degree, self-reported family economic condition, multi-variant logistic regression analysis indicated that the low and middle level of resistance to peer influence (low and middle level vs high level, had odds ratios of 2.97 (1.96 - 4.50) and 1.51 (1.05 - 2.16)), and also the middle and high level of risky decision-making (middle and high level vs low level, had odds ratios of 1.62 (1.19 - 2.22) and 3.43 (2.39 - 4.90)) were all the risk factors of adolescent health risk behaviors.

CONCLUSIONAdolescents with poor ability of resistance to peer pressure and high risky decision-making were both the risk factors of adolescent health risk behaviors.

Adolescent ; Child ; Cross-Sectional Studies ; Female ; Health Behavior ; Humans ; Logistic Models ; Male ; Peer Group ; Risk Assessment ; Risk-Taking ; Surveys and Questionnaires

Background In recent years,with the contiunous progress of the refractive surgery,the operation skill of phakic intraocular lens(PIOL)implantation for correcting extreme high myopia,astigmatism,farsightedness have made greater progression,and its security,effectiveness in clinical attract much more attention. Objective This study was to evaluate the efficacy,safety and stability of Toric Implantable Collamer Lens(TICL)for extreme high myopic astigmatism. Methods This retrospective case series included 33 eyes of 27 patients from May 2008 to February 2009.A TICL was intraocularly implanted via a 3 mm clear corneal incision after paraocular anesthesia.Patients were examined preoperatively and followed-up at 1 day,1 week,1 month,3,6,12 and 18 months postoperatively.The examinations included uncorrected visual acuity,best corrected visual acuity(BCVA),slit lamp examination,refraction,intraocular pressure,endothelial cell morphometry,etc.The written informed consent was obtained from each patient before any medical procedure. Results The uncorrected visual acuity in 96.97% eyes was equal or improved after operation in comparison with BCVA of preoperation.The spherical refraction was within-1.00 D-+0.25 D.The cylinder refraction was within-1.00 D-0 D.The axial deviation of TICL within 10 degree was 93.94%(31/33).No significant differences were found in the intraocular pressure and endothelial cell morphometry between preoperation and postoperation(intraocular pressure:F=3.35,P=5.49;endothelial cell morphometry:t=1.835,P=0.082).The visual acuity and refraction were stable during the follow-up.Astigmatic axial rotation required surgical intervention on one eye.One eye occurred high intraocular pressure because of bigger TICL diameter.The intraocular pressure returned to normal after TICL was exchanged.No cataract occurred during the follow-up duration. Conclusion TICL implantation appears to be an effective,safe and reliable method for extreme high myopic astigmatism.

Adolescent ; Eisenmenger Complex ; pathology ; therapy ; Female ; Heart-Lung Transplantation ; Humans ; Treatment Outcome