Objective To analyze serum characteristics and determine the reference range for thyroid function among healthy 11～16 year-old teenagers in Xi'an in order to offer a theoretical basis for clinical diagnosis and therapy.Methods A sum of 1 378 healthy 11～16 year-old teenagers who met the inclusion criteria from the First Affiliated Hospital of the Air Force Medical University(Xijing Hospital)between January 2020 and December 2022 were selected as research subjects,including 628 males and 750 females.They were divided into three groups based on age:Group 1:11～＜13 year-olds(433 cases),Group 2:13～＜15 year-olds(425 cases),and Group 3:15～≤16 year-olds(520 cases).Differences in serum thyroid function indices among different genders and age groups were analyzed,the reference ranges for these indices were established,and 99 healthy 11-16 year-old teenagers who met the inclusion criteria were chosen for verification.Results There were no significant differences between different genders in thyroid stimulating hormone[TSH,2.56(1.80,3.63)μIU/ml vs 2.43(1.68,3.48)μIU/ml]and total thyroxine[TT4,97.84(85.34,111.00)nmol/L vs 98.20(87.16,111.23)nmol/L],the differences were statistically significant(Z=-1.881,-0.638,all P＞0.05).Meanwhile,the differences in free thyroxine[FT4,16.93(15.49,18.60)pmol/L vs 16.26(14.80,17.83)pmol/L],free triiodothyronine[FT3,6.21(5.66,6.80)pmol/L vs 5.59(4.98,6.19)pmol/L],and total triiodothyronine[TT3,2.24(1.96,2.55)nmol/L vs 2.04(1.78,2.34)nmol/L]between different genders were significant(Z=-5.368,-11.994,-6.417 all P＜0.01).The differences in thyroid function indices were significant among different age groups(Z=10.649～261.003,all P＜0.05).The reference ranges for thyroid function indices across different age groups and genders were established,in which thyroid function indicators were verified to be within the established reference range by 99 samples.Conclusion Teenage hormone secretion varies greatly,and the secretion of thyroid hormones is influenced by various factors.Thus,the diagnosis and treatment of teenage thyroid diseases cannot fully rely on the reference ranges provided by adults or manufacturers.This study established the reference range of the thyroid function indices of 11～16 year-old teenagers in Xi'an,offering clinical doctors'diagnosis and treatment data support.

Extracellular vesicles (EVs) can carry a variety of bioactive components including nucleic acids, proteins and small molecule metabolites, and their value in tumor diagnosis and treatment has been widely recognized. However, current studies on EV inclusions mainly focus on RNA and protein, and the role of small molecule metabolites that can most directly reflect the cell state in EV remains unclear. EV metabolomics in cancer research has gradually gained traction in recent years. There are still many challenges in EV metabolomics research due to the complexity of pretreatment and low content of metabolite, but its value in regulating tumor progression and serving as tumor markers has gradually emerged, which is expected to provide new targets for tumor diagnosis and treatment.

Objective:To investigate the genetic etiology of a Marfan syndrome pedigree, and the impact of c.4336G>A variant on the splicing process of FBN1 gene.Methods:The proband was admitted to the Department of Cardiovascular Surgery of Xijing Hospital due to thoracic aortic aneurysm and dissection in August 2019. Multiplex PCR and next generation sequencing technology were used to detect 15 genes associated with hereditary aortic diseases in the proband. Then the pathogenic sites were further verified by Sanger sequencing, and above examinations were also performed among the family members of the proband. The effect of the mutation on mRNA splicing was predicted by splicing prediction software. RNAs from peripheral blood cells of the proband and the healthy person were extracted, and the effect of the mutation on mRNA splicing was verified by reverse transcription PCR and Sanger sequencing. The pathogenicity was analyzed by the recommendations from the American College of Medical Genetics (ACMG).Results:The gene panel detected a missense mutation of FBN1 gene (c.4336G>A) in the proband. Sanger sequencing results were consistent with that of panel. Sanger sequencing results showed that 4 family members were carriers of the same variant, and 3 out of the 4 family members presented signs of thoracic aortic aneurysm and dissection. The dbscSNV_ada_score and dbscSNV_rf_score software predicted that this mutation would lead to the occurrence of abnormal splicing of mRNA. The skipping of exon 35 was verified in the subsequent examinations by reverse transcription PCR and Sanger sequencing. The variant was classified as"pathogenic"according to ACMG guideline.Conclusion:FBN1 c.4336G>A mutation can cause the skipping of exon 35, and this might be the genetic mechanistic of severe cardiovascular abnormalities observed in this Marfan syndrome pedigree.

Objective:To study the clinical characteristics and death risk factors of the patients with fall-related injuries in parts of Kashgar Prefecture during 2019-2020.Methods:The clinical data were retrospectively analyzed of the 894 patients with fall-related injuries who had been admitted to Department of Trauma Center, The First People's Hospital of Kashgar Prefecture, Xinjiang Uygur Autonomous Region from January 2019 to December 2020. Recorded were the patient's gender, age, location of fall, month of fall, fall height, major injury site, injury severity score (ISS) and Glasgow score (GCS). The clinical characteristics and death risk factors of the fall patients were analyzed.Results:Of the 894 patients, 72.3%(646/894) were male and 86.9%(777/894) fell from a height from 1 to 6 meters. Their ages ranged mainly from 15 to 59 years old (74.3%, 664/894). Home was the most frequent site for falls (60.2%, 538/894) and the patients who fell in summer months (from June to August) were the most (32.3%, 289/894). Twenty-one patients (2.3%, 21/894) died. There were significant differences in the major injury site, blood transfusion, ISS score and GCS score between the dead and survival patients ( P<0.05). The multivariate logistic regression analysis showed that the injury to the head, face and neck [ OR=10.936, 95% CI: 1.177 to 101.627, P=0.035] and GCS score ≤12 [ OR=5.640, 95% CI: 2.658 to 11.968, P< 0.001] were the death risk factors for the patients with fall-related injuries in parts of Kashgar Prefecture during 2019-2020. Conclusions:In the patients with fall-related injuries in parts of Kashgar Prefecture during 2019-2020, males aged from 15 to 59 years old were the high-risk group of falls. Months with a high incidence of falls were from June to August. The fall patients with injuries to the head, face and neck and with a GCS score of ≤12 were at a high risk of death.

目的：通过 CRISPR/Cas9 技术构建前列腺癌 PC3 细胞 TFDP3 基因敲除的稳转株，探讨抑制 TFDP3 表达对 PC3 细 胞周期、凋亡、迁移和侵袭能力的影响。方法：通过生物信息学筛选 sgRNA，通过 CRISPR/Cas9 技术、构建抑制 TFDP3 基因表达 的 sgRNA-Cas9 共转染慢病毒，感染 PC3 细胞后筛选获取稳转细胞株。通过流式细胞术对 TFDP3 基因敲除的实验组与空白对照 组进行细胞周期和凋亡检测，并进一步通过划痕实验和 Transwell 实验进行细胞迁移和侵袭能力检测。结果：通过生物信息学 筛选获得 3 条 sgRNA，其中 sgRNA2 有明显的抑制前列腺癌细胞基因表达的功能；通过 CRISPR/Cas9 技术成功构建了基于 CRISPR/Cas9 介导的 TFDP3 低表达的 PC3 细胞稳转株。抑制 TFDP3 基因表达后，相比于对照组，KO 组中 G0/G1 期细胞 百分比增加、G2/M 期细胞百分比下降（P<0.05 或 P<0.01），细胞凋亡率显著升高（P<0.05），细胞迁移率明显下降 [24 h 迁移率： (44.00±1.60)% vs (65.00±4.40)%，P<0.01]，穿过聚碳酸酯膜的侵袭细胞数明显下降 [（185.89±11.71）vs （248.33±11.95）个， P<0.01]。结论：通过 CRISPR/Cas9 技术抑制 TFDP3 基因表达后，PC3 细胞发生周期阻滞、凋亡率也有所增加、迁移和侵袭能力 显著减弱，提示 TFDP3 是一个前列腺癌促癌基因。

Extracellular vesicles (EV) are widely distributed, rich in content, and fully participate in the regulation of important cellular physiological activities. As a new technology for tumor diagnosis, the natural advantages of EV include protecting its contents, being easy to obtain from various biofluids, having the potential to be truly non-invasive and high tissue fidelity. In order to transfer EV for clinical use, many isolation technologies such as microfluidics, immunochips, nano-flow cytometry and acoustic capture have been continuously developed. At present, several EV tumor diagnostic products have been applied in the clinical setting. However, due to the great differences in detection methods and platforms, there are still some technical difficulties in its clinical applications. With the accumulation of clinical evidence and the continuous improvement of clinical diagnostic methods, the liquid biopsy based on EV can finally be applied in the clinic.

As a new type of intercellular signaling rector, extracellular vesicles (EV) are involved in almost the whole process of tumorigenesis, progression, metastasis, and drug resistance. Therefore, EV have become the ideal biomarker candidates and research hotspots for cancer diagnosis and treatment. However, EV tumor biomarker research mainly focused on RNA and protein, and a small part of the research focused on lipids at the early stage. EV DNA has received little attention and its diagnostic value has gradually been recognized in recent years. Study on the biological characteristics and function of EV DNA may highlight its potential in tumor diagnosis.

As a new type of paracrine/autocrine and remote signaling, extracellular vesicles (EVs) involved in almost the whole process of tumorigenesis, progression, metastasis, and drug resistance, which has greatly inspired tumor diagnosis and treatment. EVs have the great potential to be the blood or urine biomarkers and can be used for cancer diagnosis, prognosis and monitoring. Due to its great clinical application value, EVs have become a hotspot in cancer research in recent years. Therefore, in this review, the advantages, progression, and technical challenges of EVs biomarkers in clinical tumors diagnosis are discussed.

Objective: To optimize the existing methods of isolation and purification for exosomes from urine and explore the effects of different storage conditions on the content of exosomal RNA in urine. Methods: The exosomes in human urine samples were extracted by different precipitation method, i.e., precipitation following first concentrating and direct precipitation, respectively, and the separation efficiency and cost of the two methods were compared. ExoQuick-TCTM precipitation kit was used to extract exosomes. Nanoparticle tracking analysis technique (NTA) was used to detect the concentration and particle size distribution of exosome. Dynamic light scattering (DLS) was used to detect the potential of exosome. Transmission electron microscopy (TEM) was used to observe morphology of exosomes. western blot was used to analyze the exosomal marker molecules CD63 and Alix. The extraction method of the precipitation following first concentrating was used to verify the reliability of the optimized method in 10 clinical urine samples . Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of exosomal RNA marker let-7c and PSA mRNA in the urinary exosomes from 20 patients with prostate cancer after repeated freeze-thaw (0 [i.e., fresh], 1 , 3 and 5 times) and 9 patients with prostate cancer frozen at -80 ℃ for different time (0 [i.e., fresh], 1, 2 and 4 weeks), and were statistically analyzed by Wilcoxon rank sum test for differences between the 2 groups. Results: The size distribution of exosomes extracted by the two methods was 30 to 150 nm by NTA, both of which were displayed as single peaks. The results of DLS showed that the potentials of exosome extracted by the two methods were negative values. The size of the exosomes extracted by the two methods was consistent observed under TEM namely the diameter distribution was 30 to 150 nm. western blot analysis confirmed that CD63 and Alix, the exosome labeling molecules, existed in the optimized method. The concentration of exosomes extracted from the 10 urine samples all reached 10 9 to 10 11 particles/mL. The contents of let-7c and PSA mRNA in exosomes decreased significantly after 5 freeze-thaw cycles, and the Z values were -1.79 and -1.73, respectively (P＜0.05). The RNA content of the exosomes remained stable after freezing at -80 ℃ for 1 month. Conclusion The optimized exosome extraction method could reduce greatly the cost under the premises of ensuring the concentration and quality of exosomes. The isolated exosomes may keep stable RNA content after freezing at -80 ℃ for a short time, but could not be frozen and thawed repeatedly for more than 5 times.

Objective: To investigate the effects of prostate cancer exosomes on the migration and invasion ability of stromal cells (WPMY-1), and to explore its mechanism. Methods: Exosomes in LNCaP-AI+F prostate cancer cell supernatant were isolated by ultracentrifugation and the typical structure of exosome was captured by electron microscope. The particle size distribution was analyzed by Zetaview, and Wb was used to identify the marker proteins and other proteins.After co-incubation of WPMY-1 cells and prostate cancer exosomes (40 µg/ml), laser confocal microscope was used to observe the uptake of PKH67-labeled exosomes by WPMY-1 cells; Transwell assay was used to detect the migration and invasion ability of WPMY-1 cells; qPCR was performed to detect the expression of three cancer-associated fibroblast (CAF)-related molecules (IL-8, PDGFB and MMP9) at mRNA level; and the phosphorylation of EGFR and ERK1/2 was analyzed by Wb. Results: Typical cup-shaped structure of exosomes was observed under electron microscope. The Zetaview results showed that the particle size distribution was concentrated at about 100 nm. The expression of exosome marker proteins CD63 andALIX further verified that the isolated particles were exosomes. Besides, EGFR, HER2 and SRC, which were related to the progression of prostate cancer, were also enriched in exosomes. After co-incubation, confocal microscope imaging showed a number of PKH67 labeled exosomes in recipient WPMY-1 cells. Transwell experiments showed that exosomes could significantly enhance the migration and invasion ability of WPMY-1 cells (all P<0.01). Compared with the control group, increased secretion of IL-8, PDGFB and MMP9 was observed after exosome treatment (40 µg/ml) (P<0.05 or P<0.01). Wb indicated that exosomes could promote the phosphorylation of EGFR and ERK1/2 of WPMY-1 cells (P<0.01). Conclusion: Prostate cancer cell exosomes could act on the stromal cell WPMY-1 to highly express multiple CAF-related molecules, promote the phosphorylation of EGFR and ERK1/2 and enhance the migration and invasion ability of WPMY-1 cells.