OBJECTIVEConstruct the gene library of apoptosis related genes in acute promyelocytic leukemia (APL) cell line NB4 cells treated by arsenic trioxide to clarify the apoptotic mechanism of NB4 cells.

METHODAPL cell line NB4 cells treated with or without arsenic trioxide for 24 hours. Total RNA was extracted and suppress subtractive hybridization (SSH) was conducted according to the manual. With the cDNA of the apoptosis cells as the tester and that of control cells as the driver, forward and reverse hybridization was performed. Differentially expressed genes were linked with pGEM-Teasy cloning vector and transformed into E. coli DH5alpha. The positive clones were screened by blue and white spot. PCR were used to amplify these genes.

RESULTThe subtractive cDNA libraries related with apoptosis of NB4 cells were successfully constructed.

CONCLUSIONThe constructed subtractive libraries are suitable for further study on the functional genes associated with apoptosis ofNB4 cells induced by arsenic trioxide.

Apoptosis ; drug effects ; genetics ; Arsenicals ; pharmacology ; Cell Line, Tumor ; DNA, Complementary ; analysis ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Gene Library ; Humans ; Leukemia, Promyelocytic, Acute ; pathology ; Nucleic Acid Hybridization ; methods ; Oligonucleotide Array Sequence Analysis ; Oxides ; pharmacology ; Sequence Analysis, DNA

Objective:To explore the accuracy of artificial intelligence (AI) system based on deep learning in evaluating bone age of children with abnormal growth and development.Methods:The positive X-ray films of the left wrist of children with abnormal growth and development who were treated at the Affiliated Hospital of Guizhou Medical University from January 2020 to December 2021 were collected retrospectively. A total of 717 children were collected, including 266 males and 451 females, aged 2-18 (11±3) years. Based on Tanner Whitehouse 3 (TW 3)-RUS (radius, ulna, short bone) and TW3-Carpal (carpal bone) method, bone age was measured by 3 senior radiologists, and the mean value was taken as reference standard. The bone ages were independently evaluated by the AI system (Dr.Wise bone age prediction software) and two junior radiologists (physicians 1 and 2). The accuracy within 0.5 year, the accuracy within 1 year, the mean absolute error (MAE) and the root mean square error (RMSE) between the evaluation results and the reference standard were analyzed. Paired sample t-test was used to compare MAE between AI system and junior physicians. Intraclass correlation coefficient (ICC) was used to evaluate the consistency between AI system, junior physician and reference standard. The Bland-Altman diagram was drawn and the 95% consistency limit was calculated between AI system and reference standard. Results:For TW3-RUS bone age, compared with the reference standard, the accuracy within 0.5 year of AI system, physician 1 and physician 2 was 75.3% (540/717), 62.1% (445/717) and 66.2% (475/717), respectively. The accuracy within 1 year was 96.9% (695/717), 86.3% (619/717) and 89.1% (639/717), respectively. MAE was 0.360, 0.565 and 0.496 years, and RMSE was 0.469, 0.634 and 0.572 years, respectively. For TW3-Carpal bone age, compared with the reference standard, the accuracy within 0.5 year of AI system, physician 1 and physician 2 was 80.9% (580/717), 65.1% (467/717) and 71.7% (514/717), respectively. The accuracy within 1 year was 96.0% (688/717), 87.3% (626/717) and 90.4% (648/717), respectively. MAE was 0.330, 0.527 and 0.455 years, and RMSE was 0.458, 0.612, 0.538 years, respectively. Based on TW3-RUS and TW3-Carpal bone age, the MAE of AI system were lower than those of physician 1 and physician 2, and the differences were statistically significant ( P all<0.001). The evaluation results of AI, physician 1 and physician 2 were in good agreement with the reference standard (ICC all>0.950). The Bland-Altman analysis showed that the 95% agreement limits of AI system for assessing TW3-RUS and TW3-Carpal bone age were -0.75-1.02 years and-0.86-0.91 years, respectively. Conclusion:The accuracy of AI system in evaluating the bone age of children with abnormal growth and development is close to that of senior doctors, better than that of junior doctors, and in good agreement with senior doctors.

Objective To compare the effectiveness of Defocus Incorporated Multiple Segments(DIMS)and High Aspheric Lenslet(HAL)defocus lenses in controlling myopia.Methods A total of 214 children and adolescents who were fitted with defocus lenses(DIMS or HAL)in our hospital from January to June 2023 were selected,including 100 cases in DIMS group and 114 cases in the HAL group.Changes in parameters such as spherical lens power,cylindrical lens power,spherical e-quivalent(SE),corneal curvature(K),axial length(AL),binocular accommodative response(BCC),positive/negative relative accommodation(PRA/NRA),distance/near latent phoria(DLP/NLP),and accommodative convergence to accommodation ratio(AC/A)were compared between the two groups before wearing and after wearing the lenses for 6 months,and the effectiveness of myopia control between DIMS and HAL was further compared.Results After wearing the lenses(DIMS or HAL)for 6 months,the average increase in axial length was(0.09±0.14)mm,the average in-crease in spherical lens power was(-0.11±0.23)D,the average increase in astigmatism was(-0.07±0.16)DC,and the average increase in spherical equivalent was(-0.15±0.24)D,with statistically significant differences observed for all these changes(t=9.24,-7.27,-6.60,-9.03,respectively,P＜0.001).Compared with parameters before wearing the lenses,statistically significant differences were noted after wearing for 6 months in spherical lens power,cylindrical lens power,SE,BCC,PRA,DLP,average K,and AL(P≤0.001),except for NRA(t=-0.12,P＞0.05),NLP(t=-0.28,P＞0.05),and AC/A(t=0.33,P＞0.05).The differences in parameters between the two groups after wearing the lenses for 6 months showed a statistically signifi-cant difference only in the AL difference(t=4.38,P≤0.001),with no statistically significant differences observed for other parameter differences(P＞0.05).Conclusion Wearing defocus lenses has a positive effect on controlling myopia,delaying axial length elongation,and maintaining binocular visual function balance.

Objective To compare the effectiveness of Defocus Incorporated Multiple Segments(DIMS)and High Aspheric Lenslet(HAL)defocus lenses in controlling myopia.Methods A total of 214 children and adolescents who were fitted with defocus lenses(DIMS or HAL)in our hospital from January to June 2023 were selected,including 100 cases in DIMS group and 114 cases in the HAL group.Changes in parameters such as spherical lens power,cylindrical lens power,spherical e-quivalent(SE),corneal curvature(K),axial length(AL),binocular accommodative response(BCC),positive/negative relative accommodation(PRA/NRA),distance/near latent phoria(DLP/NLP),and accommodative convergence to accommodation ratio(AC/A)were compared between the two groups before wearing and after wearing the lenses for 6 months,and the effectiveness of myopia control between DIMS and HAL was further compared.Results After wearing the lenses(DIMS or HAL)for 6 months,the average increase in axial length was(0.09±0.14)mm,the average in-crease in spherical lens power was(-0.11±0.23)D,the average increase in astigmatism was(-0.07±0.16)DC,and the average increase in spherical equivalent was(-0.15±0.24)D,with statistically significant differences observed for all these changes(t=9.24,-7.27,-6.60,-9.03,respectively,P＜0.001).Compared with parameters before wearing the lenses,statistically significant differences were noted after wearing for 6 months in spherical lens power,cylindrical lens power,SE,BCC,PRA,DLP,average K,and AL(P≤0.001),except for NRA(t=-0.12,P＞0.05),NLP(t=-0.28,P＞0.05),and AC/A(t=0.33,P＞0.05).The differences in parameters between the two groups after wearing the lenses for 6 months showed a statistically signifi-cant difference only in the AL difference(t=4.38,P≤0.001),with no statistically significant differences observed for other parameter differences(P＞0.05).Conclusion Wearing defocus lenses has a positive effect on controlling myopia,delaying axial length elongation,and maintaining binocular visual function balance.

Objective To establish a PCR-based capillary electrophoresis(PCR/CE)to detect Survival Motor Neuron 1(SMN1)and Survival Motor Neuron 2(SMN2)genes and to evaluate its performance.Methods PCR/CE and Multiplex Ligation-dependent Probe Amplification(MLPA)for SMA gene diagnosis were used to blindly test the samples in sync.The performance of PCR/CE was assessed using MLPA results as the standard.Results A total of 336 samples were included in this study,consisting of 50 homozygous deletion types(14.9%),65 heterozygous deletion types(19.3%),and 221 non-deletion types(65.8%).The results of PCR/CE for detect-ing SMN1 and SMN2 copy numbers(0,1,2,3,≥4)were in complete agreement with the results of the MLPA.Conclusions PCR/CE for gene testing related to SMA could accurately detect copy numbers of exon 7 and exon 8 of the SMN1 and SMN2 genes(0,1,2,3,≥4).

【Objective】 To analyze the impact of sporadic cases of COVID-19 on the work of Transfusion Department, so as to explore the countermeasures. 【Methods】 The admission of inpatient departments, the reception of outpatient(including emergency) departments, the workload of transfusion department(including blood typing, unexpected antibody screening and cross matching), and the consumption of blood components in the Xijing Hospital between October and November in 2021, during COVID-19 outbreak, were collected. All the above data was statistically compared to the data in same period in 2018, before the COVID-19 outbreak. 【Results】 Due to the COVID-19 epidemic, there was a significant decrease in number of inpatients(280±157.1 vs 340.4±110.2), outpatient(including emergency)(8 359±3 615 vs 10 151±3 225), the workload of blood typing(272.0±132.4 vs 341.6±110.4), unexpected antibody screening(78.26±42.22 vs 98.51±43.53) and crossmatch(237.2±99 vs 475.7±155.6), as well as the consumption(U) of all blood components(457.9±50.32 vs 579.4±62.51) in the Xijing Hospital(P<0.05). In detail, the epidemic had the most direct impact on the number of inpatients and outpatients, which shrank continuously on the 2^nd day after official announcement of the new COVID-1 cases. While the workload of blood typing, unexpected antibody screening and crossmatch decreased slightly, with a lag, usually on the 2^nd, 3^rd and 5^th day after official announcement. The decrease of the usage of red blood cells and plasma began from the 7^th day after the new epidemic to the 6^th day after the end of the epidemic. However, the usage platelets and cryoprecipitate coagulation factors decreased from the 8^th and 10^th day after the new epidemic to the 2^nd and 6^th day after the end of the epidemic, respectively. 【Conclusion】 The daily work of Blood Transfusion Department has been seriously affected by sporadic COVID-19 epidemic. The working mode, staff structure and inventory ratio of blood components should be adjusted and optimized instantly to maintain the normal conduct of medical treatments in hospitals and ensure the safety of patients.

Objective:To explore the clinical application of preimplantation genetic testing for monogenic disorders (PGT-M) in an unique case with Spinal muscular atrophy (SMA) type 2+ 0.Methods:A special SMA family presented at the Third Affiliated Hospital of Guangzhou Medical University on October 19, 2020 was selected as the study subject. Multiple ligation-dependent probe amplification (MLPA) and molecular tagging linkage analysis were carried out to identify the SMN1 genotype of the couple and their fetus. Subsequently, next-generation sequencing (NGS), molecular tagging linkage analysis, and chromosomal microarray analysis were employed to determine the haplotypes and validate the result of PGT-M on the 11 embryos derived for the couple. Results:The female partner was identified as a carrier of the rare SMN1[2+ 0] variant, and prenatal diagnosis confirmed the fetus to be affected by SMA. Ultimately, PGT-M has successfully selected four embryos free from the pathogenic SMN1 variants and X chromosome deletion. Conclusion:PGT-M can effectively prevent the transmission of rare genetic variants such as the SMA 2+ 0 subtype in the families. Above finding has provided guidance for genetic counseling and family planning for the couple.

Two proteins of similar molecular weight (named as ASPR-C-1 and ASPR-C-2) from the crude drug of Angelica sinensis were purified and characterized by 80% ammonium sulfate precipitation, Sephadex G-50 gel filtration chromatography, and DEAE-Sepharose anion exchange chromatography. The molecular weight of ASPR-C-1 and ASPR-C-2 on SDS-PAGE was 17.33 kDa and 17.18 kDa, respectively. They were mainly monomeric in solution, but partially formed dimers and they were glycoproteins with glycosyl content of 2.6% and 8.2%, respectively. Both ASPR-C-1 and ASPR-C-2 were identified to be members of pathogenesis-related 10 family of proteins by matrix-assisted laser desorption ionization time-of-flight mass spectrometry and have ribonuclease activities with the specific activity of 73.60 U/mg and 146.76 U/mg, respectively. The optimum pH of the two isoforms was similar, at about 5.6, while their optimum temperatures were different. The optimum temperature of ASPR-C-1 was 50 ℃, and that of ASPR-C-2 was 60 ℃. Both isoforms presented highest thermal stability at 60 ℃. However, ASPR-C-2 was more thermotolerant than ASPR-C-1. The latter was rapidly inactivated and retained only about 20% residual activity while the former still maintained about 80% of its original activity at a higher treatment temperature (80 to 100 ℃). In addition, Fe²⁺ had an activating effect on the ribonuclease activities of two isoforms while Ca²⁺, Mg²⁺, Zn²⁺, Mn²⁺, Ag⁺, Cu²⁺, EDTA (Elhylene diamine tetraacetic acid), dithiothreitol and sodium dodecylsulphate showed different degrees of inhibition of the enzyme activities. Our findings provide a foundation for further research on the biological function of PR-10 protein from Angelica sinensis.
Angelica sinensis ; Chromatography, Gel ; Chromatography, Ion Exchange ; Electrophoresis, Polyacrylamide Gel ; Enzyme Stability ; Hydrogen-Ion Concentration ; Kinetics ; Molecular Weight ; Protein Isoforms ; Temperature