Clinical characteristics of 11 patients with chronic lymphocytic leukemia with t (14;19) (q32;q13) / 中华血液学杂志

Objective: To analyze the clinicopathological characteristics of 11 cases of chronic lymphocytic leukemia (CLL) with t (14;19) (q32;q13) . Methods: The case data of 11 patients with CLL with t (14;19) (q32;q13) in the chromosome karyotype analysis results of the Blood Diseases Hospital, Chinese Academy of Medical Sciences from January 1, 2018, to July 30, 2022, were retrospectively analyzed. Results: In all 11 patients, t (14;19) (q32;q13) involved IGH::BCL3 gene rearrangement, and most of them were accompanied by +12 or complex karyotype. An immunophenotypic score of 4-5 was found in 7 patients and 3 in 4 cases. We demonstrated that CLLs with t (14;19) (q32;q13) had a mutational pattern with recurrent mutations in NOTCH1 (3/7), FBXW7 (3/7), and KMT2D (2/7). The very-high-risk, high-risk, intermediate-risk, and low-risk groups consisted of 1, 1, 6, and 3 cases, respectively. Two patients died, 8 survived, and 2 were lost in follow-up. Four patients had disease progression or relapse during treatment. The median time to the first therapy was 1 month. Conclusion: t (14;19) (q32;q13), involving IGH::BCL3 gene rearrangement, is a rare recurrent cytogenetic abnormality in CLL, which is associated with a poor prognosis.

Molecular features of 109 patients with chronic myelomonocytic leukemia in a single center / 中华血液学杂志

Objective: To explore the molecular features of chronic myelomonocytic leukemia (CMML) . Methods: According to 2022 World Health Organization (WHO 2022) classification, 113 CMML patients and 840 myelodysplastic syndrome (MDS) patients from March 2016 to October 2021 were reclassified, and the clinical and molecular features of CMML patients were analyzed. Results: Among 113 CMML patients, 23 (20.4%) were re-diagnosed as acute myeloid leukemia (AML), including 18 AML with NPM1 mutation, 3 AML with KMT2A rearrangement, and 2 AML with MECOM rearrangement. The remaining 90 patients met the WHO 2022 CMML criteria. In addition, 19 of 840 (2.3%) MDS patients met the WHO 2022 CMML criteria. At least one gene mutation was detected in 99% of CMML patients, and the median number of mutations was 4. The genes with mutation frequency ≥ 10% were: ASXL1 (48%), NRAS (34%), RUNX1 (33%), TET2 (28%), U2AF1 (23%), SRSF2 (21.1%), SETBP1 (20%), KRAS (17%), CBL (15.6%) and DNMT3A (11%). Paired analysis showed that SRSF2 was frequently co-mutated with ASXL1 (OR=4.129, 95% CI 1.481-11.510, Q=0.007) and TET2 (OR=5.276, 95% CI 1.979-14.065, Q=0.001). SRSF2 and TET2 frequently occurred in elderly (≥60 years) patients with myeloproliferative CMML (MP-CMML). U2AF1 mutations were often mutually exclusive with TET2 (OR=0.174, 95% CI 0.038-0.791, Q=0.024), and were common in younger (<60 years) patients with myelodysplastic CMML (MD-CMML). Compared with patients with absolute monocyte count (AMoC) ≥1×10(9)/L and <1×10(9)/L, the former had a higher median age of onset (60 years old vs 47 years old, P<0.001), white blood cell count (15.9×10(9)/L vs 4.4×10(9)/L, P<0.001), proportion of monocytes (21.5% vs 15%, P=0.001), and hemoglobin level (86 g/L vs 74 g/L, P=0.014). TET2 mutations (P=0.021) and SRSF2 mutations (P=0.011) were more common in patients with AMoC≥1×10(9)/L, whereas U2AF1 mutations (P<0.001) were more common in patients with AMoC<1×10(9)/L. There was no significant difference in the frequency of other gene mutations between the two groups. Conclusion: According to WHO 2022 classification, nearly 20% of CMML patients had AMoC<1×10(9)/L at the time of diagnosis, and MD-CMML and MP-CMML had different molecular features.

Analysis of genotypes on 850 newborns with SLC26A4 single-allele mutation and the phenotypes of those with second variant / 中华耳鼻咽喉头颈外科杂志

Objective: To clarify the phenotypes of the newborns with SLC26A4 single-allele mutation in deafness genetic screening and second variant; to analyze the SLC26A4 genotype and hearing phenotype. Methods: 850 newborns born in Beijing from April 2015 to December 2019 were included and there were 468 males and 382 females. They received genetic deafness screening for 9 or 15 variants, with the result of SLC26A4 single-allele mutation. Firstly, three step deafness gene sequencing was adopted in this work, i.e., the first step was "SLC26A4 gene whole exons and splice sites" sequencing; the second step was "SLC26A4 gene promoter, FOXI1 gene and KCNJ10 gene whole exons" sequencing; and the third step was detection for "SLC26A4 gene copy number variation". Secondly, we collected the results of newborn hearing screening for all patients with the second mutation found in the three step test, and conducted audiological examinations, such as acoustic immittance, auditory brainstem response and auditory steady state response. Thirdly, for novel/VUS mutations, we searched the international deafness gene database or software, such as DVD, ClinVar and Mutation Taster, to predict the pathogenicity of mutations according to the ACMG guideline. Lastly, we analyzed the relationship between genotype and phenotype of newborns with SLC26A4 single allele mutation. Results: Among 850 cases, the median age of diagnosis was 4 months. In the first step, 850 cases were sequenced. A total of 32 cases (3.76%, 32/850) of a second variants were detected, including 18 cases (2.12%, 18/850) with identified pathogenic variants; 832 cases were sequenced and 8 cases of KCNJ10 gene missense variants were detected among the second step. No missense mutations in the FOXI1 gene and abnormal SLC26A4 gene promoter were detected; the third step sequencing results were all negative. Genotypes and hearing phenotypes included 18 cases combined with the second clear pathogenic variant, 16 cases (16/18) referred newborn hearing screening and 2 cases (2/18) passed in both ears; degree of hearing loss consisted of 18 profound ears (18/36), 13 severe ears (13/36) and 5 moderate ears (5/36); audiogram patterns comprised 17 high frequency drop ears (17/36), 14 flat ears (14/36), 3 undistinguished ears (3/36), and 2 U shaped ears (2/36); 11 cases underwent imaging examination, all of which were bilateral enlarged vestibular aqueduct. As for 22 cases of other genotypes, all passed neonatal hearing screening and the hearing diagnosis was normal, including 9 cases with VUS or possibly novel benign variants, 8 cases with KCNJ10 double gene heterozygous variants, and 5 cases with double heterozygous variants. Conclusions: The probability of individuals with SLC26A4 single-allele variant who merge with a second pathogenic variant is 2.12%, all of which are SNV, which can provide scientific basis for the genetic diagnosis and genetic counseling of SLC26A4 variants. Those who have merged with second pathogenic variant are all diagnosed with sensorineural hearing loss. Patients with KCNJ10 gene mutations do not manifest hearing loss during the infancy, suggesting the need for further follow-up.

Efficacy and safety of endoscopic diaphragm incision in children with congenital duodenal diaphragm / 中华儿科杂志

Objective: To explore the efficacy and safety of endoscopic diaphragm incision in pediatric congenital duodenal diaphragm. Methods: Eight children with duodenal diaphragm treated by endoscopic diaphragm incision in the Department of Gastroenterology of Guangzhou Women and Children's Medical Center from October 2019 to May 2022 were enrolled in this study. Their clinical data including general conditions, clinical manifestations, laboratory and imaging examinations, endoscopic procedures and outcomes were retrospectively analyzed. Results: Among the 8 children, 4 were males and 4 females. The diagnosis was confirmed at the age of 6-20 months; the age of onset was 0-12 months and the course of disease was 6-18 months. The main clinical manifestations were recurrent non-biliary vomiting, abdominal distension and malnutrition. One case complicated with refractory hyponatremia was first diagnosed with atypical congenital adrenal hyperplasia in the endocrinology department. After treatment with hydrocortisone, the blood sodium returned to normal, but vomiting was recurrent. One patient underwent laparoscopic rhomboid duodenal anastomosis in another hospital but had recurred vomiting after the operation, who was diagnosed with double duodenal diaphragm under endoscope. No other malformations were found in all the 8 cases. The duodenal diaphragm was located in the descending part of the duodenum, and the duodenal papilla was located below the diaphragm in all the 8 cases. Three cases had the diaphragm dilated by balloon to explore the diaphragm opening range before diaphragm incision; the other 5 had diaphragm incision performed after probing the diaphragm opening with guide wire. All the 8 cases were successfully treated by endoscopic incision of duodenal diaphragm, with the operation time of 12-30 minutes. There were no complications such as intestinal perforation, active bleeding or duodenal papilla injury. At one month of follow-up, their weight increased by 0.4-1.5 kg, with an increase of 5%-20%. Within the postoperative follow-up period of 2-20 months, all the 8 children had duodenal obstruction relieved, without vomiting or abdominal distension, and all resumed normal feeding. Gastroscopy reviewed at 2-3 months after the operation in 3 cases found no deformation of the duodenal bulbar cavity, and the mucosa of the incision was smooth, with a duodenal diameter of 6-7 mm. Conclusion: Endoscopic diaphragm incision is safe, effective and less invasive in pediatric congenital duodenal diaphragm, with favorable clinical applicability.

Optimization of production process of Aeromonas veronii CA07 strain inactivated vaccine / 中国生物制品学杂志

@#Abstract：Objective To optimize the production process of inactivated vaccine of Aeromonas veronii（AV）CA07 strain. Methods The fermentation culture process of AV CA07 strain liquid was determined through the optimization of the culture time（2～16 h），medium（optimized fermentation medium，LB medium and NB medium）and fermentation conditions（in⁃ oculation amount of 1%，5%，10% and 15%；ventilation rate of 2，4，6 and 8 L／min and fermentation time of 6，8，10 and 12 h）. The optimal inactivation process was determined through the comparison of the final concentration of formalde⁃ hyde solution（0. 10%，0. 20%，0. 30% and 0. 40%），inactivation temperature（28 and 37 ℃）and inactivation time（24， 48 and 72 h）. The large⁃scale production process of inactivated vaccine of AV CA07 strain in 500 L fermentor was estab⁃ lished and the prepared vaccines were tested for safety and immunogenicity. Results The optimal inoculation amount of AV CA07 strain was 5%，ventilation rate was 4 L／min and culture time was 10 ~ 12 h. The optimal inactivation condition was adding formaldehyde solution with final concentration of 0. 30% incubating at 37 ℃ for 24 h. The number of viable bacteria in the fermentation broth of AV CA07 strain prepared in 500 L fermentor was more than 8 × 109 CFU／mL. All crucian carps immunized with the inactivated vaccine by abdomen survived. After challenge，the relative immune protection rate was more than 90%. Conclusion AV CA07 strain inactivated vaccine prepared by optimized production process showed good safety and immunogenicity.

Secretory phospholipase A2 responsiveness and in vitro anti-tumor activity of oxaliplatin-loaded liposomes modified with facial amphiphiles / 中国药科大学学报

@#Modulating drug release from liposomes at tumor sites are important for eliciting therapeutic effects of platinum drugs considering their low permeability through liposomal membranes, here a novel secretory phospholipase A2 (sPLA2) responsive-liposome system was constructed for oxaliplatin (L-OHP).Lipid ingredients dipalmitoyl phosphatidylcholine and distearoyl phosphoethanolamine-PEG2k, together with facial amphiphiles (FAs) including lithocholic acid (LCA) or 3-keto lithocholic acid (kLCA) were used to prepare sPLA2 responsive-liposome (LCA-Lip or kLCA-Lip) by thin-film hydration method.The physicochemical properties, sPLA2-responsive drug release and anti-tumor activity were evaluated in vitro.The results indicated L-OHP loaded liposomes modified with FAs had similar particle sizes of approximately 100 nm and narrow size distributions (PDI < 0.11).Compared with non-FAs-containing liposomes (C-Lip), LCA-Lip or kLCA-Lip has a comparable entrapment efficiency and loading efficiency.LCA-Lip or kLCA-Lip didn't show significant higher drug leakage at the presence of 10% or 50% fetal bovine serum (FBS) in media than that in media without FBS.Treated with secretory phospholipase A2 from Colo205 cells culture conditioned medium (CCM sPLA2) for 24 h, FAs modified liposomes released about 70% of carboxyfluorescein (CF), while C-Lip only released 20% of CF.Compared to L-OHP loaded C-Lip, L-OHP-loaded FAs-included formulations had much greater anti-proliferative activity against sPLA2-secreting Colo205 cells.In summary, our results shows that LCA or kLCA promotes responsiveness of liposomes to tumor-related sPLA2 and points to a new way to develop platium drugs-loaded liposomal delivery systems with better release mechanisms.

Effective Antiviral Activity of the Tyrosine Kinase Inhibitor Sunitinib Malate against Zika Virus

Background@#(Introduction): Zika virus (ZIKV), a mosquito-borne flavivirus, causes the outbreaks of Latin America in 2015 - 2016, with the incidence of neurological complications. Sunitinib malate, an orally bioavailable malate salt of the tyrosine kinase inhibitor, is suggested as a broadspectrum antiviral agent against emerging viruses like severe acute respiratory syndrome coronavirus (SARS-CoV) and SARS-CoV-2. @*Materials and Methods@#This study investigated the antiviral efficacy and antiviral mechanisms of sunitinib malate against ZIKV infection using cytopathic effect reduction, virus yield, and time-of-addition assays. @*Results@#Sunitinib malate concentration-dependently reduced ZIKV-induced cytopathic effect, the expression of viral proteins, and ZIKV yield in supernatant with 50% inhibitory concentration (IC 50 ) value of 0.015 μM, and the selectivity index of greater than 100 against ZIKV infection, respectively. Sunitinib malate had multiple antiviral actions during entry and post-entry stages of ZIKV replication. Sunitinib malate treatment at entry stage significantly reduced the levels of ZIKV RNA replication with the reduction of (+) RNA to (-) RNA ratio and the production of new intracellular infectious particles in infected cells. The treatment at post-entry stage caused a concentration-dependent increase in the levels of ZIKV (+) RNA and (-) RNA in infected cells, along with enlarging the ratio of (+) RNA to (-) RNA, but caused a pointed increase in the titer of intracellular infectious particles by 0.01 and 0.1 μM, and a substantial decrease in the titer of intracellular infectious particles by 1 μM. @*Conclusion@#The study discovered the antiviral actions of sunitinib malate against ZIKV infection, demonstrating a repurposed, host-targeted approach to identify potential antiviral drugs for treating emerging and global viral diseases.

Analysis of genotypes and hearing phenotypes of mutation infants with deafness / 临床耳鼻咽喉头颈外科杂志

Objective@#The aim of this study is to explore the genotype and hearing phenotype of deaf infants with mutation of GJB2 gene.@*Method@#Subjects were 121 infants with GJB2 gene mutations who were treated in the Children's Hearing Diagnosis Center of Beijing Tongren hospital. All subjects were accepted to undertake the universal newborns hearing screening（UNHS） and series of objective audiometry, including auditory brainstem response, distortion product otoacoustic emission, auditory steady-state response and other audiological tests. All subjects were screened for nine pathogenic variants in four genes or all exons of the GJB2 gene, and then were diagnosed as infants with GJB2 gene mutations. Initially, analyzing their genotypes and hearing phenotypes generally. Then, the subjects were divided into two groups according to the genotypes: T/T group（truncated/truncated mutations, 89 cases） and T/NT group（truncated/non-truncated mutations, 32 cases）. Chi-square test was used to analyze the results of UNHS, hearing degree, audiogram patterns and symmetry/asymmetry of binaural hearing phenotype. Eventually, analyzing the results of UNHS. @*Result@#The most common truncated mutation was c.235delC（64.88%, 157/242） and the most common non-truncated mutation was c.109G>A（11.16%, 27/242）. The homozygous mutation of c.235delC/c.235delC was the dominant in T/T group（38.84%, 47/121）, and the compound heterozygous mutation of c.235delC/c.109G>A was the dominant in T/NT group（18.18%, 22/121）. 81.82%（99/121） of subjects failed in UNHS, including 74.38%（90/121） with bilateral reference, 7.44%（9/121） with a single pass. The refer rate of UNHS of group T/T and T/NT were 86.52%（77/89） and 68.75%, respectively. There was a statistically significant difference between the two groups（P<0.05）. 85.95%（104/121） of subjects were diagnosed as hearing loss and 14.05%（17/121） of subjects were diagnosed as normal hearing. The degree of hearing loss: profound, severe, moderate and mild were 31.40%（38/121）, 19.01%（23/121）, 24.79%（30/121） and 10.74%（13/121）, respectively. There was no subjects with normal hearing in T/T group and individuals with severe and profound hearing loss accounted for the highest proportion（65.17%, 58/89）, while in T/NT group, normal hearing accounted for 53.13%（17/32） and mild and moderate hearing loss accounted for the highest proportion（37.5%, 12/32）. There was statistically significant difference between the two groups（P<0.05）. Of 104 patients（208 ears） with hearing loss, the audiogram patterns: flat, descending, ascending, residual, Valley and other types were 49.03%（102/208）, 12.02%（25/208）, 8.65%（18/208）, 7.69%（16/204）, 3.36%（7/204） and 19.23%（40/204）, respectively. The two most common types in T/T group were flat（47.19%, 84/178） and other types（20.22%, 36/178）, while in T/NT group were flat（60.00%, 18/30） and ascending（20.00%, 6/30）. There was statistically significant difference between the two groups（P<0.05）. There were 50 cases（48.07%） with symmetrical hearing phenotype and 54 cases（51.93%） with asymmetrical hearing phenotype. Asymmetry was predominant in T/T group（53.93%, 48/89）, and symmetry was predominant in T/NT group（60.00%, 9/15）. There was no statistically significant difference between the two groups（P>0.05）. @*Conclusion@#In this study, c.235delC/c.235delC homozygous mutation was dominant in T/T group and c.235delC/c.109G>A heterozygous mutation was dominant in T/NT Group. The hearing phenotypes in T/T group were mostly bilateral asymmetric severe hearing loss, and those in T/NT Group were bilateral symmetric mild to moderate hearing loss, special attention should be paid to the audiological characteristics of different genotypes.

Analysis of genotypes and hearing phenotypes of mutation infants with deafness / 临床耳鼻咽喉头颈外科杂志

The aim of this study is to explore the genotype and hearing phenotype of deaf infants with mutation of gene. Subjects were 121 infants with gene mutations who were treated in the Children's Hearing Diagnosis Center of Beijing Tongren hospital. All subjects were accepted to undertake the universal newborns hearing screening（UNHS） and series of objective audiometry, including auditory brainstem response, distortion product otoacoustic emission, auditory steady-state response and other audiological tests. All subjects were screened for nine pathogenic variants in four genes or all exons of the gene, and then were diagnosed as infants with gene mutations. Initially, analyzing their genotypes and hearing phenotypes generally. Then, the subjects were divided into two groups according to the genotypes: T/T group（truncated/truncated mutations, 89 cases） and T/NT group（truncated/non-truncated mutations, 32 cases）. Chi-square test was used to analyze the results of UNHS, hearing degree, audiogram patterns and symmetry/asymmetry of binaural hearing phenotype. Eventually, analyzing the results of UNHS. The most common truncated mutation was c.235delC（64.88%, 157/242） and the most common non-truncated mutation was c.109G>A（11.16%, 27/242）. The homozygous mutation of c.235delC/c.235delC was the dominant in T/T group（38.84%, 47/121）, and the compound heterozygous mutation of c.235delC/c.109G>A was the dominant in T/NT group（18.18%, 22/121）. 81.82%（99/121） of subjects failed in UNHS, including 74.38%（90/121） with bilateral reference, 7.44%（9/121） with a single pass. The refer rate of UNHS of group T/T and T/NT were 86.52%（77/89） and 68.75%, respectively. There was a statistically significant difference between the two groups（0.05）. In this study, c.235delC/c.235delC homozygous mutation was dominant in T/T group and c.235delC/c.109G>A heterozygous mutation was dominant in T/NT Group. The hearing phenotypes in T/T group were mostly bilateral asymmetric severe hearing loss, and those in T/NT Group were bilateral symmetric mild to moderate hearing loss, special attention should be paid to the audiological characteristics of different genotypes.

Cis-3-O-p-hydroxycinnamoyl Ursolic Acid Induced ROS-Dependent p53-Mediated Mitochondrial Apoptosis in Oral Cancer Cells

Cis-3-O-p-hydroxycinnamoyl ursolic acid (HCUA), a triterpenoid compound, was purified from Elaeagnus oldhamii Maxim. This traditional medicinal plant has been used for treating rheumatoid arthritis and lung disorders as well as for its anti-inflammation and anticancer activities. This study aimed to investigate the anti-proliferative and apoptotic-inducing activities of HCUA in oral cancer cells. HCUA exhibited anti-proliferative activity in oral cancer cell lines (Ca9-22 and SAS cells), but not in normal oral fibroblasts. The inhibitory concentration of HCUA that resulted in 50% viability was 24.0 µM and 17.8 µM for Ca9-22 and SAS cells, respectively. Moreover, HCUA increased the number of cells in the sub-G1 arrest phase and apoptosis in a concentration-dependent manner in both oral cancer cell lines, but not in normal oral fibroblasts. Importantly, HCUA induced p53-mediated transcriptional regulation of pro-apoptotic proteins (Bax, Bak, Bim, Noxa, and PUMA), which are associated with mitochondrial apoptosis in oral cancer cells via the loss of mitochondrial membrane potential. HCUA triggered the production of intracellular reactive oxygen species (ROS) that was ascertained to be involved in HCUA-induced apoptosis by the ROS inhibitors YCG063 and N-acetyl-L-cysteine. As a result, HCUA had potential antitumor activity to oral cancer cells through eliciting ROS-dependent and p53-mediated mitochondrial apoptosis. Overall, HCUA could be applicable for the development of anticancer agents against human oral cancer.

Effect of ambient temperature on determination of carbohydrate antigen 242 by Luminex flow cytometry fluorescence immunoassay / 第二军医大学学报

[Abstract] Objective To analyze the difference of the carbohydrate antigen 242 (CA242) results measured by Luminex flow cytometry fluorescence immunoassay with different calibrations and different ambient temperatures, so as to evaluate the effect of ambient temperature on the measurement results. Methods Two CA242 samples with different concentrations (median- and high-value) were selected and the determination was repeated at 11 temperature points (18.3, 20.1, 21.5, 22.2, 24.1, 25.0, 25.4, 26.5, 27.4, 28.5, and 30.0 ℃) for 5 times by Luminex flow cytometry fluorescence immunoassay. The average value of the detection result at the calibration temperature (25 ℃) was taken as the reference value, and the biases of the detection results at different ambient temperatures were calculated. Forty-nine CA242 specimens covering high-, median- and low-values were examined at 20, 25 and 30 ℃ by Luminex flow cytometry fluorescence immunoassay, and the results were compared and correlation analysis was carried out. Results The biases of the median- and high-value samples at 18.3 ℃ were －35.6% and －29.4%, respectively. The difference between the measurement results at 20 ℃ and 25 ℃ was significant for the 49 specimens (P0.001), and there was a linear correlation between them (Y＝0.676 9X＋0.374 7, R2＝0.990 5). The difference of measurement results was significant between 30 ℃ and 25 ℃ (P0.001), and there was a linear correlation between them (Y＝0.896 6X＋0.227 0, R2＝0.999 4). Conclusion Too low or too high ambient temperature has a negative effect on the determination of CA242 by Luminex flow cytometry fluorescence immunoassay. The ambient temperature should be kept relatively constant and controlled within the range of calibration temperature ±4 ℃.

Clinical Features and Therapeutic Efficacy in Adult Acute Lymphoblastic Leukemia with t (1; 19) (E2A-PBX1) / 中国实验血液学杂志

OBJECTIVE@#To explore the clinical features and therapeutic efficacy in adult ALL patients with t (1; 19) (E2A-PBX1).@*METHODS@#The clinic data of 19 adult ALL patients with t (1; 19) (E2A-PBX1) in our hospital from Nov. 22, 2010 to Apr. 4, 2018 were collected. The clinical features,complete remission (CR) rate, overall survival (OS) rate and relapse-free survival (RFS) rate of patients received chemotherapy and chemotherapy+HSCT were analyzed.@*RESULTS@#In all the 19 patients, the median age was 24 (14-66), median WBC count was 16.47×109 (1.8-170.34)/L, median Hb level was 98 (65-176) g/L, median Plt count was 50 (15-254)×109/L. Pre B-ALL were 17 cases (89.5%), and common B-ALL were 2 cases (10.5%). Patients received the induction therapy, the overall CR rate was 94.7%, one course CR rate was 94.7%, 4 year OS rate was 47.1% and RFS rate was 43.3%. The OS rate and RFS rate of patients received transplantation were slightly higher than those of patients not received transplantation (OS: 62.5% vs 36.7%) (P=0.188)；RFS (62.5% vs 38.9%) (P=0.166).@*CONCLUSION@#Most adult ALL patients with t (1; 19) (E2A-PBX1) is Pre B-ALL by Immunophenotyping, as compared with the pediatric patients, the therapeutic efficacy for adult patients with t (1; 19) (E2A-PBX1) is worsen, therefore, stem cell transplantation is still acquired for better long term survival.

Clinical Characteristics of Patients with Ph Mixed Phenotype Acute Leukemia / 中国实验血液学杂志

OBJECTIVE@#To investigate the clinical biological characteristics and prognosis of the patients with mixed phenotype acute leukemia with t(9;22)(q34;q11.2) and/or BCRABL1 (Ph MPAL).@*METHODS@#The morphological, immunological, cytogenetic, and molecular features of 33 in patients with Ph MPAL were retrospectively analyzed in our center from June 2002 to June 2016 according to the scoring proposal of European Group for the Classification of Acute Leukemia(EGIL )1998 and WHO 2008 criteria. All the cases were either treated with acute lymphoblastic leukemia (ALL) induction regimen or combined chemotherapy regimens for both acute lymphoblastic and acute myeloid leukemia,part of which also received tyrosine kinase inhibitor(TKI) and 5 cases underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT) after complete remission.@*RESULTS@#Ph MPAL occurred predominantly in male patients (ratio of M/F was 1.75∶1), and a high WBC counts at diagnosis; the WBC count was higher than 30×10/L in 25 patients( 75.8% )， and appeared higher than 100 ×10/L in 13 patients ( 39.4%). Among all the 33 PhMPAL patients, 32 (97.0%) had a myeloid / B-lymphoid (M/B) phenotype， and 1 case(3.0%) had a myeloid/ B-lymphoid/ T-lymphoid/ (M/B/T) phenotype. There was no patients displayed myeloid / T-lymphoid (M/T) or B-lymphoid/ T-lymphoid/ (B/T) phenotype. 19 of all cases(57.6%) met the diagnosis criteria of PhMPAL based on EGIL 1998 criteria, while the remaining 14 cases can be diagnosed as Ph MPAL by WHO 2008 classification,but excluded as PhMAPL by EGIL 1998.Karyotype analysis was successfully performed in 31 cases, and out of them 13 (41.9%) had a sole Ph chromosome, 10 (32.3%) had additional chromosome aberration and Ph chromosome was not found in 8 cases (25.8%) .In 31 patients the fusion gene BCR/ABL (P190、P210) was detected,including 17 (54.8%) cases with the p190 BCR/ABL transcript, 8 (25.8%) cases with the p210 BCR/ABL transcript, 4 (12.9%) expressing both transcripts and 2 (6.5%) without any one of these 2 transcripts. 24 out of 33 patients (77.4%) achieved complete remission after induction therapy. The median time achieving CR was 43(26-98)days. The CR rate of patients treated with and without imatinib after the first inducion treatment was 81.3% and 46.7%,respectively (P0.05). Within the 17 patients treated with imatinib at induction stage，2 of which became BCR/ABLnegative．At consolidation chemotherapy stage, 9 out of 16 patients became BCR/ABL negative, including 3 patients already subjected to HSCT. The median time reached to BCR/ABL negative was 2.87（1.13-9.20）months.@*CONCLUSION@#Ph MPAL is more common in male, and inclined to high WBC counts at diagnosis. Myeloid/B lymphoid phenotype is more common, and the prognosis of patients with PhMPAL is poor. Imatinib and allogeneic hematopoietic stem cell transplantation may improve survival of patients with PhMPAL.

Evaluation value of echocardiographic index and serum myocardial enzyme level changes for gestational hypertension／ / 心血管康复医学杂志

To explore relationship among disease severity , serum myocardial enzyme levels and echocar‐diographic indexes in patients with gestational hypertension (GH).Methods : A total of 101 GH inpatients treated in our department of gynecology from 2015 to 2018 were divided into severe group (severe preeclampsia and eclampsia patients , n＝46 ) and mild group (mild preeclampsia patients , n＝ 55 ) , and another 60 normal pregnant women were simultaneously selected as normal control group .Heart indexes , serum levels of creatine kinase isoenzyme MB (CK‐MB) and creatine kinase (CK ) were measured and compared among three groups .Results : Compared with normal control group , there were significant rise in LVESd , LVEDd , right ventricular diameter (RVD) , end‐di‐astolic interventricular septal thickness (IVSTd ) and left ventricular posterior wall thickness (LVPWT ) in mild group and severe group , and above indexes of severe group were significantly higher than those of mild group , P＜0.01 all.Compared with normal control group , there was significant rise in transmitral late diastolic peak flow ve‐locity (A) , and significant reductions in LVEF , cardiac output (CO) , cardiac index (CI) , transmitral early dias‐tolic peak flow velocity (E) and E／A in mild group and severe group ;compared with mild group , there was signifi‐cant rise in A , and significant reductions in LVEF , CO , CI , E and E／A in severe group , P＜0. 01 all .Compared with normal control group , there were significant rise in serum levels of CK‐MB [ (6.2 ± 2.5) IU／L vs.(9.4 ± 3.2) IU／L vs.(15. 3 ± 5. 7) IU／L] and CK [(41.8 ± 7.9) IU／L vs.(61.7 ± 12. 5) IU／L vs.(88. 7 ± 20. 4) IU／L] in mild group and severe group , and those of severe group were significantly higher than those of mild group , P＝0.001 all.Conclusion : Echocardiography and myocardial enzyme examination in GH patients help to understand functional state and structure of heart , which possess important significance for early identification and treatment .

Expression of oleosin-rhFGF9 fusion protein in Carthamus tinctorius and determination of hair regeneration and wound repair potential in mice / 中国中药杂志

The expression of fibroblast growth factor 9 (FGF9) recombinant fusion protein in Carthamus tinctorius was used to identify its effect on hair regrowth and wound repair system in mice, providing a basis for C. tinctorius as a plant bioreactor, and establishing a foundation for commercial applications of FGF9 fusion protein in hair regrowth and wound repair. The identified pOTBar-oleosin-rhFGF9 plasmid was transformed into Agrobacterium tumefaciens EHA105 by freeze-thaw method, and the oleosin-rhFGF9 gene was transformed into safflower leaves by A. tumefaciens mediated method. Transgenic safflower seedlings were then obtained by tissue culture. After basta screening, transgenic T₃ safflower seeds were obtained by grafting method, PCR verification and propagation. The expression of oleosin-rhFGF9 was detected by Western blot, and the content of oleosin-rhFGF9 fusion protein was 0.09% by using ELISA quantitative method. It was observed that 60 μg·L⁻¹ transgenic safflower oil had better effect on promoting NIH/3T3 cells proliferation in a certain dose-dependent manner. Sixty C57BL/6 mice were used to establish alopecia model and wound model respectively, and then were randomly divided into control group (treated with PBS or saline), negative group (treated with wild type safflower seed oil bodies, 60 g·L⁻¹), positive group (treated with FGF9, 0.054 g·L⁻¹), low dose group (treated with transgenic safflower oil bodies, 10 g·L⁻¹) and high dose group (treated with transgenic safflower oil bodies, 60 g·L⁻¹). The skin of all above-mentioned mice models were coated with soft adhesive manner every other day, 100 μL/time. After 15 days, the mice skin was cut and embedded for histological analysis. The hair regrowth experimental results showed that the hair of mice grew well, and the mice in high dose group had bushy hair, with significant effect on regeneration hair number as compared with the positive group. The healing was obvious in wound experiment, with significant healing effect in positive group, high dose group and low dose group as compared to blank control group. Furthermore, high dose group remarkably showed a better and higher healing effect than the positive group at day 5. Oleosin-rhFGF9 was successfully transformed into safflower, and T₃ transgenic safflower oil bodies expressed oleosin-rhFGF9 fusion protein were obtained, with the role of promoting hair regeneration and wound repair in mice.

Dietary Restriction Reduces Blood Lipids and Ameliorates Liver Function of Mice with Hyperlipidemia / 华中科技大学学报（医学）（英德文版）

Dietary restriction (DR) can delay senescence,prolong lifespan of mammals and improve their learning-memory activity.The purpose of the study was to explore the effects of DR on hypolipidemic action and liver function of mice with hyperlipidemia.To investigate these effects,hyperlipidemia mouse models were established with high-fat diet (HFD) (34％ of energy),then randomly divided into HFD group,DR30％ group and DR50％ group.Mice in DR30％ and DR50％ group were respectively supplied with HFD as much as about 70％ and 50％ of the consumption of HFD in the mice of HFD group.Rats in control group were fed routinely.After DR for 5 weeks,the average body weight,liver weight,liver index,serum lipids and glucose levels in both DR groups decreased significantly as compared with the HFD group (P＜0.05 or P＜0.01),so did alanine aminotransferase (ALT),aspartate aminotransferase (AST),lactate dehydrogenase (LDH) levels and the ratio of LDL-C/HDL-C in the DR50％ group (P＜0.05 or P＜0.01).Histopathology examination of liver tissues further proved ameliorative effect of DR on liver function.Western blotting showed that DR significantly increased the expression of silent mating type information regulation 2 homolog 1 (SIRT1) in liver and adipose,while notably decreased the expression of peroxisome proliferators-activated receptors-gamma (PPARγ) in adipose (P＜0.05 or P＜0.01).The increase of SIRT1 and decrease of PPARγ may be a mechanism by which DR reduces blood lipids and ameliorates liver function.

Clinical and Cytogenetic Characteristics of Two AML Patients with High-level MLL Expression / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the clinical and cytogenetic characteristics of high-level mixed-lineage leukaemia (MLL) gene amplification in patients with acute myeloid leukemia (AML).</p><p><b>METHODS</b>The clinical and cytogenetic data of 2 AML patients with high-level MLL amplification from January 2010 to August 2016 were analyzed retrospectively.</p><p><b>RESULTS</b>The two AML cases were in middle-aged population. They were diagnosed as FAB subtype M5b and M2a respectively. Both of them had complex karyotypes with the aberrations of chromosome 11. One case was confirmed as MLL-PTD involving exons 2-9 by RT-PCR and sequencing. The other case without MLL-PTD was further analyzed by CytoScan HD analysis. The CMA results showed partial gain of 11q accompanied with partial loss in 11q, deletion of regions in 3p, 3q, 4q, 5q, 7q, 8q, 10p, 10q, 12p and 18q, as well as gain of 4p.</p><p><b>CONCLUSION</b>The co-existence of -5/5q-, -7/7q- and highly complex karyotype may accelerate the poor prognosis. Thus how those cytogenetic abnormalities influencing the disease prognosis need to be further explored.</p>

Characteristics and Prognostic Significance of Cytogenetic Abnormalities in Diffuse Large B-Cell Lymphoma Patients with Bone Marrow Involvement / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the cytogenetic abnormalitis in patients with diffuse large B-cell lymphoma(DLBCL) patients with bone marrow involvement and their influence on prognosis.</p><p><b>METHODS</b>Conventional karyotyping was performed on bone marrow specimens in 47 DLBCL patients with histologically confirmed bone marrow involvement(BMI). The karyotyping results of bone marrow, the characteristics and clinical effect of chromosomal abnormalities were analysed.</p><p><b>RESULTS</b>In 47 DLBCL cases with BMI, the chromosomal abnormalities were detected in 25(53%) cases. Among them, complex karyotype was more frequent, being noted in 19(40%) patients. The most frequently involved chromosomes were chromosome 1 and 18(both 26%), others were chromosome 3(23%), 6(19%), 7, 8 and 14(13%). Among all karyotype changes, the most common numerical aberrations, in decreasing order of incidence, were trisomy 3(13%), trisomy 5, trisomy 7, trisomy 12, trisomy 18 and loss of 21(6%,each), and the most predominant structural aberrations, in decreasing order of incidence, were 1q+(17%), 1p+, 6q-, 8q+, 14q+, 18p+, 18q+ and aberrations involving band 2p21-p23 (6%,each). The prognostic impact analysis of both clinical features and cytogenetic aberrations revealed that IPI≥3 (P=0.03) or the presence of chromosomal abnormalities (P=0.005) were significantly related with poor progression free survival(PFS), and IPI≥3 (P=0.024), lactate dehydrogenase(LDH)≥ three times of the upper limit of normal (P=0.027) and the presence of chromosomal abnormalities (P=0.001) predominantly related with poor overall survival(OS). In multivariate analysis, the presence of chromosomal abnormalities was the only independently adverse factor for PFS(P=0.037, HR 2.323) and OS(P=0.015, HR 2.833). The analysis of prognostic effects of specific chromosomal aberrations showed that patients with specific cytogenetic abnormalities of 1q+, 8q+, +12, 12q+, 18p+ and aberrations involving band 2p21-23 had significantly poor PFS, and patients with specific cytogenetic abnormalities of 1q+, +3, +5, +7, 8q+, +12, 12q+ and aberrations involving band 2p21-23 had significantly poor OS. When the above mentioned specific chromosomal aberrations were analyzed with clinical covariate, the presence of chromosomal aberration of 8q+ (P=0.022, HR 2.701) and IPI≥3 (P=0.043, HR 2.949) were independently poor prognostic factors for PFS, and 1q+ (P=0.032, HR 2.973) was the independently poor prognostic factor for OS.</p><p><b>CONCLUSION</b>In DLBCL patients with BMI, the presence of chromosomal abnormalities is the only independently poor factor for PFS and OS, and among them, the specific cytogenetic aberrations of 8q+ or 1q+ have an independently poor prognostic impact on PFS or OS, respectively, which need to be further studied.</p>

Reduced Autophagy in 5-Fluorouracil Resistant Colon Cancer Cells

We investigated the role of autophagy in SNUC5/5-FUR, 5-fluorouracil (5-FU) resistant SNUC5 colon cancer cells. SNUC5/5-FUR cells exhibited low level of autophagy, as determined by light microscopy, confocal microscopy, and flow cytometry following acridine orange staining, and the decreased level of GFP-LC3 puncta. In addition, expression of critical autophagic proteins such as Atg5, Beclin-1 and LC3-II and autophagic flux was diminished in SNUC5/5-FUR cells. Whereas production of reactive oxygen species (ROS) was significantly elevated in SNUC5/5-FUR cells, treatment with the ROS inhibitor N-acetyl cysteine further reduced the level of autophagy. Taken together, these results indicate that decreased autophagy is linked to 5-FU resistance in SNUC5 colon cancer cells.

Optimized Exon-Exon Junction Library and its Application on Rodents' Brain Transcriptome Analysis

ABSTRACT Background: Alternative splicing (AS), which plays an important role in gene expression and functional regulation, has been analyzed on genome-scale by various bioinformatic approaches based on RNA-seq data. Compared with the huge number of studies on mouse, the AS researches approaching the rat, whose genome is intermedia between mouse and human, were still limited. To enrich the knowledge on AS events in rodents' brain, we perfomed a comprehensive analysis on four transcriptome libraries (mouse cerebrum, mouse cerebellum, rat cerebrum, and rat cerebellum), recruiting high-throughput sequencing technology. An optimized exon-exon junction library approach was introduced to adapt the longer RNA-seq reads and to improve mapping efficiency. Results: In total, 7,106 mouse genes and 2,734 rat genes were differentially expressed between cerebrum and cerebellum, while 7,125 mouse genes and 1,795 rat genes exhibited varieties on transcript variant level. Only half of the differentially expressed exon-exon junctions could be reflected at gene expression level. Functional cluster analysis showed that 32 pathways in mouse and 9 pathways in rat were significantly enriched, and 6 of them were in both. Interestingly, some differentially expressed transcript variants did not show difference on gene expression level, such as PLCβ1 and Kcnma1. Conclusion: Our work provided a case study of a novel exon-exon junction strategy to analyze the expression of genes and isoforms, helping us understand which transcript contributes to the overall expression and further functional change.