OBJECTIVETo investigate the time course of let-7a microRNA expression in the cell cycle of HeLa cells.

METHODSHeLa cells were synchronized in G(1), S and G(2)/M phases using double-thymidine block, and the cell cycle phases were defined by flow cytometry. Real-time quantitative RT-PCR was used to examine the expression of let-7a in HeLa cells in different cell cycle phases.

RESULTSThe synchronization rates of G(1), S and G(2)/M phases were 84.81%, 83.65% and 77.69%, respectively. Let-7a was constitutively expressed throughout the cell cycle in HeLa cells, but the expression levels in G(1) and S phases were lower than those in G(2)/M phase.

CONCLUSIONSCell cycle can significantly influence the expression level of let-7a, which may provide new clues to the understanding of the cell cycle control mechanisms.

Cell Cycle ; genetics ; Gene Expression Regulation, Neoplastic ; physiology ; HeLa Cells ; Humans ; MicroRNAs ; genetics ; metabolism

Objective To observe the effects of estradiol benzoate (E_2),human cberionic gonadotropin (hCG) and luteinizing hormone-releasing hormone (LHRH) on testicular descent and their relation to the expression of insulin-like factor-3 (INSL3) mBNA in rats.Methods At birth,40 SD male rats were assigned to 4 groups,and each group consisted of 10 rats.Each group received injection of one of the following drugs:normal saline(NS),E_3,hCG+E_2,and LHRH+E_2 for 29 clays after birth.Total RNA was extracted from the testis on day 30 after birth.INSL3 was synthesized and further amplified by RT-PCR.Results (1) Testicular descent rate on clay 30 after birth was 100% (20/20) in the group of NS,0% (0/20) in the group of E_2,85% (17/20) of hCG + E_2,and 70% (14/20) of LHBH + E_2.There was no significant difference between the last two groups ( P = 0.45).(2) Weight of testis differed significantly among the groups ( F = 56.67,P＜0.01 ),being lightest in the group of E_2.(3) E_2 could damage the germ cells of testis,however hCG and LHRH ameliorated this situation. (4) The expression of INSL3 mBNA was absent in the group of E_2,while the other groups showed normal expression.Conclusion E_2 inhibits testicular descent in rats,while hCG and LHRH promote the progress.The mechanism seems to be related to the expression of INSL3 mRNA.

OBJECTIVE: To compare the agreement of anterior chamber angle examination by ultrasound biomicroscope (UBM), slit lamp optical coherence tomography (SL-OCT), and gonioscopy in angle recession and angle closure. METHODS: The anterior chamber angle was measured with UBM, SL-OCT and gonioscopy in turns for temporal, nasal, superior and inferior quadrant in the same dark room. The results were compared with the agreement of the three methods in angle recession and angle closure by χ2 test and Kappa test. RESULTS: There were no statistically significant differences of the three methods in testing angle closure and angle recession (P>0.05). The consistency of UBM and gonioscopy was better (Kappa value of 0.882) than that of SL-OCT and gonioscopy (Kappa value of 0.624). CONCLUSION When testing angle recession, UBM is better than SL-OCT with gonioscopy as the standard. When testing angle closure, UBM, SL-OCT and gonioscopy have good agreement.
Anterior Chamber ; Glaucoma, Angle-Closure ; Gonioscopy ; Humans ; Intraocular Pressure ; Microscopy, Acoustic ; Slit Lamp ; Tomography, Optical Coherence

OBJECTIVETo investigate the effect of microRNA on the gene expression profile of human leukemia K562 cells using microarray technique.

METHODSmiR-181a RNA duplexes were designed and synthesized according to the mature sequence of miR-181a. Forty-eight hours after transfection of in vitro cultured K562 cells using Oligofectamine, gene expression profiles of the cells were studied and analyzed using Agilent Human 1A Oligo microarray.

RESULTSTotalling 228 differentially expressed genes were identified from the 20,173 screened genes, including 59 up-regulated ones (consisting of metabolism-associated genes, tumor suppressor genes, signal transduction-associated genes, immunity and defense-associated genes etc), and 169 down-regulated ones (consisting of oncogenes, DNA-binding and transcription genes, metabolism-associated genes, signal transduction-associated genes, cell cycle and development-associated genes etc.) in the transfected K562 cells as compared with the control K562 cells. Changes in expressions of CTCF, ZAP70, SEMA4C and RALA were confirmed by semi-quantitative reverse transcription-polymerase chain reaction.

CONCLUSIONSmiR-181a transfection for 48 h induces gene expression profile changes in K562 cells, indicating the functionality of the miR-181a. These differentially expressed genes are related to the functions of the microRNA, and may also be the basis of the regulation model of posttranscriptional gene silencing. These findings provide an evidence for further study of the machineries and functions of the microRNA in mammalian cells.

Gene Expression Profiling ; Humans ; K562 Cells ; MicroRNAs ; genetics ; Oligonucleotide Array Sequence Analysis ; RNA, Small Interfering ; genetics ; Transfection

Objective To observe the current status of secondary prevention medication usage and their relation with on-treatment platelet reactivity in patients with Acute Coronary Syndrome(ACS) treated with aspirin and clopidogrel. Methods A total of 176 patients hospitalized from 2014 to 2015 due to ACS in the Department of Cardiology, Peking University People's Hospital were enrolled and on-treatment platelet reactivity was tested by thromboelastography(TEG)and CYP2C19*2,*3 and*17 alleles were analysed. Details of secondary prevention medication and patients' clinical characteristics were recorded. The relation of secondary prevention medication and on-treatment platelet reactivity was analyzed by multi-logistic regression after adjusting for CYP2C19 alleles and clinical characteristics covariates.Results A 94.89% of patients was treated with statins while 80.68% with beta blocker. The platelet inhibition rate were (45.33±28.78)% and the high on-treatment platelet reactivity (HTPR) rate tested by TEG was 37.50%. In the multivariate logistic regression analysis, usage of β-blockers during hospitalization as well as phenotypes of CYP2C19*2,*3 and *17,clinical presentation with ST-segment elevation myocardial infarction and the length of stents were associated with HTPR defi ned by TEG. The percentage of HTPR rate was signifi cantly lower in patients treated with than those without β-blockers (72.73% vs. 85.45%,OR 0.18,95%CI 0.06-0.53,P=0.002)after adjusting genetic factors and other covariates.Conclusions There was a signifi cant correlation between beta blockers usage and high clopidogrel on-treatment platelet reactivity.

Objective To investigate the therapeutic effect and safety of ESE (endoscopic submucosalexcavation) as the derivative technology of ESD (endoscopic submucosal dissection) for intestinal Neuroendocrinetumor. Methods A total of 23 lesions diagnosed as Neuroendocrine tumor were treated by ESE. Pathologicaldiagnosis was performed. Reverse events were recorded.Patients were followed up for recurrence andmetastasis. Results Lesions, 0.4 ~ 3.0 cm (medium size 1.0 cm) in diameter,were all resected at one ESE procedure.The operation time was 20 ~ 75 min (medium 30 min). Postoperative bleeding occurred in one case .Initiative fullthickness resection was made in one case due to the violation of muscularis propria layer. 23 cases were histologicallydiagnosed as neuroendocrine tumor, with 21 as G1 and 2 as G2, none as G3. Within 3 gastric neuroendocrine tumors,2 were type 1 and 1 was type 2. All resected samples were free of residual tumor cell in the lateral and basalmargins. Conclusion ESE is safe and efficacious for the treatment of intestinal neuroendocrine tumor.

Objective:To compare and analyze the clinical diagnostic values of five thyroid nodule malignant risk stratification guidelines.Methods:From October 2019 to October 2021, 926 cases of patients with 1 027 thyroid nodules were recruited in the Second Affiliated Hospital of Xi ′an Jiaotong University. All nodules were categorized individually according to 2015 American Thyroid Association for Ultrasound Malignancy Risk Stratification of Thyroid Nodules in Adults Guidelines(ATA guidelines), 2016 the Korean Society of Radiology and the Korean Society of Thyroid Radiology Thyroid Imaging Reporting and Data Systems(K-TIRADS), 2017 European Thyroid Association Thyroid Imaging Reporting and Data Systems(Eu-TIRADS), 2017 American College of Radiology Thyroid Imaging Reporting and Data Systems (ACR TI-RADS), and 2020 Chinese Thyroid Imaging Reporting and Data System (C-TIRADS). The pathological results were taken as the gold standard, the malignancy ratio of nodules of different categories in each system was calculated. ROC curves were plotted to evaluate the diagnostic efficiencies of different systems, and DeLong test was used to compare the areas under ROC curves. The sensitivity and specificity of different systems were calculated when the maximum point of the Youden index was the optimal cut-off value.Results:In the same stratified system, there were statistically significant differences in the malignant proportion of nodules of different grades ( P<0.05). The malignant proportion of nodules in the high-risk group showed no significant difference among different stratified systems ( P>0.05). Except for C-TIRADS, the malignant proportion of nodules was increased with the increase of diagnostic grade at each grade of the other four stratification systems. ROC curve showed that AUCs of ATA guidelines, K-TIRADS, EU-TIRADS, ACR TI-RADS and C-TIRADS were 0.814, 0.819, 0.814, 0.820 and 0.802, respectively, there was no statistical significance in AUC of different stratification systems (all P>0.05). The optimal truncation values in differentiating benign and malignant nodules were middle-risk malignant nodules, moderately suspicious malignant nodules, middle-risk malignant nodules, class 4 and class 4B. The diagnostic of five stratification systems showed that ATA guidelines had the highest sensitivity (0.784), C-TIRADS had the highest specificity (0.854). Conclusions:The five stratified systems have similar efficacy in differentiating benign and malignant thyroid nodules, and all of them have good diagnostic value.

Objective:To explore the clinical and imaging characteristics of mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS) caused by mitochondrial DNA 14453G>A (m.14453G>A) mutation.Methods:A case of MELAS caused by m.14453G>A mutation in the First Affiliated Hospital of Harbin Medical University on October 12, 2021 was reported. At the same time, the reported cases of MELAS and Leigh syndrome (LS) caused by the m.14453G>A mutation were reviewed. This enabled a comprehensive summarization, analysis, and comparison of these cases.Results:The patient was a female. She has suffered from the disease since 13-year old with seizures, accompanied by the disturbance of mood and the loss of memory. Brain magnetic resonance imaging findings consisted of lesions in frontal, parietal, occipital, temporal lobe and cerebellar. The patient was initially considered with autoimmune encephalitis and posterior reversible encephalopathy syndrome. Since direct sequencing of the complete mitochondrial genome from blood of the patient revealed m.14453G>A mutation in ND6 gene, and the mutation rate was 17.0%, the patient eventually diagnosed with MELAS based on clinical manifestations, imaging examinations, and genetic testing results. Using "m.14453G>A" as the search term, the relevant literature in China and abroad was retrieved and those with complete clinical data were identified. A total of 11 cases of m.14453G>A mutation including this case were reported, of whom 5 patients were diagnosed as MELAS, and 6 patients were diagnosed as LS. Among the 11 patients, those being adolescent or adult and with lesions in the cortex and subcortical white matter were probably be MELAS; those being infant or young child and with lesions in basal ganglia, thalamus and brainstem could be LS. Conclusions:Mitochondrial disease caused by m.14453G>A gene mutation shows a great heterogeneity, which can cause MELAS and LS. The clinical phenotype of the m.14453G>A mutation may be related to the age of onset and lesion′ s location.

Thoracic obstruction is mainly attributed to the scope of coronary heart disease in modern medicine, and traditional Chinese medicine(TCM) shows a significant effect in the treatment of thoracic obstruction. In this research, a network pharmacology method was carried out to systemically study the underlying mechanism of the core herbal compatibility in TCM on the thoracic obstruction. First, we collected the literature about TCM prescriptions for treating thoracic obstruction from CNKI. Then, a prescription database was establish by TCM inheritance support platform system(V2.5) to determine the medication rules and core herbal compatibility in TCM. Finally, to obtain the potential signaling pathways, KEGG pathway analysis was performed by BATMAN-TCM online analysis tool. Results showed that the potential signal pathway of core herbal compatibility in TCM for the clinical treatment of thoracic obstruction was calcium ion and cGMP-PKG signaling pathway. This study provided a new research strategy for the study of the medication rules and mechanism of traditional Chinese medicine in the treatment of thoracic obstruction.

BACKGROUNDThis study aimed to evaluate the effects of standard rescue procedure (SRP) in improving severe trauma treatments in China.

METHODSThis study was conducted in 12 hospitals located in geographically and industrially different cities in China. A standard procedure on severe trauma rescue was established as a general rule for staff training and patient treatment. A regional network (system) efficiently integrating prehospital rescue, emergency room treatments, and hospital specialist treatments was built under the rule for information sharing and improving severe trauma treatments. Treatment outcomes were compared between before and 1 year after the implementation of the SRP.

RESULTSThe outcomes of a total of 74,615 and 12,051 trauma cases were collected from 12 hospitals before and after the implementation of the SRP. Implementation of the SRP led to efficient cooperation and information sharing of different treatment services. The emergency response time, prehospital transit time, emergency rescue time, consultation call time, and mortality rate of patients were 24.24 ± 4.32 min, 45.69 ± 3.89 min, 6.38 ± 1.05 min, 17.53 ± 0.72 min, and 33.82% ± 3.87% (n = 441), respectively, before the implementation of the standardization and significantly reduced to 10.11 ± 3.21 min, 22.39 ± 4.32 min, 3.26 ± 0.89 min, 3.45 ± 0.45 min, and 20.49% ± 3.11%, separately (n = 495, P < 0.05) after that.

CONCLUSIONSStaff training and SRP can significantly improve the efficiency of severe trauma treatments in China.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; Emergency Medical Services ; standards ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Wounds and Injuries ; Young Adult