A retrospective analysis of the clinical data of seven acute B-lymphoblastic leukemia (B-ALL) patients with TCF3-HLF fusion gene-positive admitted to the First Affiliated Hospital of Soochow University from June 2017 to August 2022 was conducted to summarize their clinical features and prognoses. The seven B-ALL patients comprised four males and three females, with a median age of 18 (11-33) years. Five patients tested positive for CD33 expression, and four patients had a normal karyotype. Two patients had hypercalcemia at the initial diagnosis, and one patient developed hypercalcemia at relapse. Six patients presented with coagulation dysfunction at diagnosis. After induction chemotherapy, five out of seven patients achieved complete remission, of which four subsequently relapsed. Two patients did not achieve remission even after two rounds of induction chemotherapy, with one achieving complete remission after treatment with blinatumomab immunotherapy. Three patients underwent chimeric antigen receptor T cell therapy, whereas three patients subsequently underwent hematopoietic stem cell transplantation. Five patients died, while two patients survived with sustained complete remission. TCF3-HLF-positive B-ALL is rare and has a high relapse rate and poor prognosis.

Objective:To compare the assay and related substance detection methods of erythromycin lactobionate in the Chinese Pharmacopoeia 2020(ChP),USP 2023(USP),JP18(JP),BP2023(BP)and EP 11.0(EP),investigate the differences between the test results obtained from 7 batches of erythromycin lactobionate for injection samples by using ChP and BP methods and thus provide a reference for the improvement of specification of erythromycin lactobionate.Methods:The differences of test methods and limits under the items of assay and related substances of erythromycin lactobionate in the above five pharmacopoeias were listed and compared.The related substances and contents of erythromycin lactobionate for injection samples from different manufactures were tested with methods stated in ChP and BP,and then compared and analyzed.Results:The items of related substances were determined by high performance liquid chromotographic methods in the ChP,EP and BP.The test methods and limits in the EP and BP were the same,which were different from that in ChP.The related substances were not determined in the USP and JP.There are obvious differences between the chromatographic methods and limits for the items of related substances in the ChP and BP,i.e.,BP contains 7 specific impuri-ties,while ChP contains only 2 specific impurities.The limits for any other impurity and the total amount of impurities were lower in the BP than those in the ChP.The antibiotic microbiological assay was used as the test method for the item of assay in the ChP,USP and JP based on different bacterial strains,which was different from the chromatographic method used in the BP.Based on the different methods of the ChP and BP,the related substances and content determination results of 7 batches of erythromycin lactobionate for injection samples met the acceptance criteria.The detection efficiency of BP related substances inspection method for specific impurities and total impurities were significantly higher than that of the ChP method.Conclusion:The BP method is superi-or to the ChP method in the detection of erythromycin lactobionate and erythromycin lactobionate for injection related substances.In terms of content determination,the external standard method adopted by BP is feasible to replace the antibiotic microbiological assay adopted by the ChP.Related substances and content determination methods adopted by the BP can provide an important reference for the revision and improvement of erythromycin lactobionate standard in the ChP.

Taking the micro-course The Connection Between Vertebrae as an example, this paper discusses the teaching design, production process and practical application of the anatomy micro-course, analyzes the existing problems and puts forward corresponding suggestions. The micro-course design integrates knowledge points with ideological and political elements, and applies a variety of teaching methods to make the teaching content interesting, enlightening and applicable. In the production process, using anatomical specimens, models and 3D software demonstration structure can make abstract knowledge intuitive and perceptual. Moreover, the artistry of micro-course is increased appropriately, and the misunderstanding of emphasizing appearance and neglecting design is avoided. The application of micro-course integrates the micro-course and online education platform to realize the intelligent education of information teaching. In addition, college teachers should improve the awareness of micro-course production and carry out systematic construction and application of micro-course resources.

Late onset methylmalonic acidemia (MMA) is a rare genetic metabolic disease.This case is a 46 year old adult patient with MMA complicated with hyperhomocysteinemia.It starts with progressive limb weakness and mental abnormality, and has dysuria and respiratory failure.Neurological examination showed decreased muscle strength of limbs and pyramidal tract sign.The levels of blood homocysteine and urinary methylmalonic acid increased significantly.Head, neck, thoracolumbar magnetic resonance imaging showed abnormal signals in the spinal cord from the level of foramen magnum to the level of lumbar 1 vertebral body.Two heterozygous variants of mmachc were found by gene detection: c: 609G>A, c: 349G>A, consistent with cobalamin C deficiency.Treat with L-carnitine, vitamin B12 and betaine.The patients′ mental symptoms, limb muscle strength and respiratory failure were improved, and the level of blood homocysteine also decreased significantly.

Objective:Xp11.22 microduplication syndrome is a very rare disease.In July 2017, 2 male patients with Xp11.22 microduplication syndrome of the same family were admitted to the 980th Hospital of the PLA Joint Service Support Force.Diagnosis process: the medical exons of the proband and his parents were sequenced by high-throughput sequencing technology, and the gene sequences were compared and analyzed.Genomic copy number variation of proband, his uncle and his mother were analyzed by chromosome microarray.Medical exon sequencing did not find gene mutations that were highly correlated with the clinical phenotype of the proband.The results of chromosome microarray analysis showed that the proband had a 629 kb fragment duplication in the chrXp11.22 region, and the gene locus was Xp11.22 (53, 188, 779-53, 817, 598), which contained HSD17B10, HUWE1, SMC1A, KDM5C and IQSEC2 important OMIM genes, associated with Xp11.22 microduplication syndrome, it was a pathogenic copy number change.The same 612Kb fragment duplication in the chrXp11.22 region, locus Xp11.22 (53, 188, 779-53, 800, 670) was found in his uncle.And the same 803Kb fragment duplication in the chrXp11.22 region, locus (53, 188, 779-53, 991, 495) was found in his mother.In the families with unknown intelligence, especially male patients, it is necessary to detect the whole genome copy number of the patients to be alert to Xp11.22 microrepeat syndrome.

Objective To explore the genetic basis for a newborn infant suspected with Donohue syndrome.Methods Whole exome sequencing (WES) was used to screen potential variants in the child.Suspected variants were validated through Sanger sequencing and real-time PCR.Results The child was found to carry two heterozygous variants in the INSR gene,including c.3258+4(IVS17)A＞G and deletion of exon 2,which were respectively inherited from her mother and father.Conclusion The compound heterozygous variants of the INSR gene probably underlie the disease in this patient.

Objective: To investigate the predictive value of serum high-mobility group box-1 protein (HMGB1) for hemorrhage transformation (HT) after intravenous thrombolysis in patients with acute ischemic stroke. Methods: From February 2017 to September 2019, patients with acute ischemic stroke underwent intravenous thrombolysis in Lixin County People's Hospital, Bozhou, Anhui Province were enrolled prospectively. In the morning of the day after admission, fasting blood was collected to detect the level of serum HMGB1. Twenty-four hours after intravenous thrombolysis, CT reexamination was performed to determine whether HT occurred. The demographic and baseline clinical data were compared between the HT group and the non-HT group. Multivariate logistic regression analysis was used to determine the independent risk factors for HT after thrombolysis. Receiver operating characteristic (ROC) curve was used to analyze the predictive value of serum HMGB1 level to HT. Results: A total of 182 patients were enrolled in the study, including 22 in the HT group and 160 in the non-HT group. The age, fasting blood glucose, serum HMGB1 level, and the proportion of history of atrial fibrillation and regular antiplatelet medication before onset in the HT group was significantly higher than those in the non-HT group, and the differences were statistically significant (all P<0.05). Multivariate logistic regression analysis showed that the increased serum HGMB1 level (odds ratio [OR] 2.145, 95% confidence interval[CI] 1.467-3.138; P=0.002), taking antiplatelet drugs regularly before onset (OR 5.496, 95% CI 1.700-17.768; P=0.004) and increased baseline fasting blood glucose level (OR 1.333, 95% CI 1.024-1.736; P=0.033) were the independent risk factors for HT after intravenous thrombolysis. ROC curve analysis showed that the area under the curve of serum HMGB1 level predicting HT after intravenous thrombolysis was 0.788 (95% CI 0.721-0.845; P<0.001). The sensitivity and specificity were 72.73% and 82.50%, respectively, when the best cutoff value was 7.97 μg/L. Conclusion The increased baseline HMGB1 level may predict the risk of HT after intravenous thrombolysis in patients with acute ischemic stroke.

OBJECTIVE: To analyze the clinical manifestation and genetic mutation of a child with tyrosinemia type I but without elevated succinylacetone. METHODS: Clinical data of the patient was collected. Tandem mass spectrometry and gas chromatography mass spectrometry were used to analyze the blood amino acid and urine organic acid component of the proband. DNA was extracted from the child and his parents and used for mutation analysis. RESULTS: The proband was of acute type, with features including hepatomegaly, jaundice, anemia and tendency of bleeding. Serum levels of Tyrosine, Methionine and Phenylalanine were 397.12 μmol/L, 896.16 μmol/L and 292.52 μmol/L, respectively, which all distinctly exceeded the normal levels. The level of phenyllactic acid and 4-hydroxyphenyl-lactic acid of proband's urine were 17.4 μmol/L and 417.0 μmol/L, respectively, which also exceeded the normal levels, but the level of succinylacetone was within the normal range. Compound heterozygous mutations of the FAH gene, namely c.634delT (p.L212Wfs*20) and c.455G>A (p.W152X), were detected in the proband, which were both predicted to be pathogenic and were inherited from her father and mother, respectively. CONCLUSION For children with tyrosinemia type I, detection of urine succinylacetone by gas phase mass spectrometry can be negative. The diagnosis of tyrosinemia type I must rely on genetic testing and/or enzymatic assaying.
DNA Mutational Analysis ; Female ; Genetic Testing ; Heptanoates ; Humans ; Male ; Tyrosinemias

OBJECTIVE： To prepare Ganshen granules， formulate its quality standards primarily and establish its HPLC fingerprint. METHODS： Using feeding speed， roller speed， roller pressure and roller clearance as factor， grain forming rate as index， single factor test and orthogonal test were used to optimize the granulation technology of Ganshen granules. According to 2015 edition of Chinese Pharmacopeia （part Ⅳ） （shorted for pharmacopeia）， moisture， granulation and dissolution were determined. TLC was used for the qualitative identification of Lycium barbarum， Astragalus membranaceus， Codonopsis pilosula in the Ganshen granules. HPLC method was used to determine the contents of betaine， calycosin-7-glucoside and lobetyolin in Ganshen granules. Fingerprints of 10 batches of Ganshen granules were drawn. RESULTS： The optimal dry granulation technology of Ganshen granules included that 25 r/min feeding speed， 8 r/min roller speed， 7 MPa roller pressure and 1.1 mm roller clearance， The grain forming rate is 85.83％. The moisture， granulation and solubility of Ganshen granule were all in line with pharmacopeia standard. TLC of L. barbarum， A. membranaceus and C. pilosula showed the same color spots on the corresponding positions of the reference chromatogram. The linear range of sample mass of betaine is 4.32-8.64 μg， and the linear range of mass concentration of calycosin-7-glucoside and lobetyolin were 5-30 and 10-60 μg/mL， respectively. RSDs of precision， reproducibility and stability tests （24 h） were all lower than 2.0％ （n＝5）. Average recoveries were 97.02％， 99.25％ and 101.04％ （all RSD＜1.7％， n＝6 or n＝9）. The contents of them were 4.298、0.054、0.025 mg/g， respectively. The similarity of HPLC fingerprints of 10 batches of Ganshen granules to control fingerprint was higher than 0.95. CONCLUSIONS： The optimal granulation technology of Ganshen granule is stable and feasible， and established quality standard and HPLC fingerprint can provide reference for quality control of Ganshen granule.

Osteosarcoma is a clinically common primary malignant bone tumor.Its treatment is still not ideal because of the high de-gree of malignancy and early metastasis.Understanding the molecular mechanism and the biological characteristics of malignant os-teosarcoma,and exploring the effective treatment have become the focus of osteosarcoma research at home and abroad.A long non-coding RNA,lncRNA,is involved in the regulation of a variety of cellular functions and plays an important role in the development of osteosarcoma.Recently,lncRNA has been increasingly reported in osteosarcoma research.It has been found to affect the proliferation, invasion,and migration of osteosarcoma cells by interacting with proteins,mRNAs,and miRNAs.This article reviews the research on ln-cRNA based on recent reports regarding its role in the molecular mechanism of osteosarcoma.