1.A Study of Flow Sorting Lymphocyte Subsets to Detect Epstein-Barr Virus Reactivation in Patients with Hematological Malignancies.
Hui-Ying LI ; Shen-Hao LIU ; Fang-Tong LIU ; Kai-Wen TAN ; Zi-Hao WANG ; Han-Yu CAO ; Si-Man HUANG ; Chao-Ling WAN ; Hai-Ping DAI ; Sheng-Li XUE ; Lian BAI
Journal of Experimental Hematology 2025;33(5):1468-1475
OBJECTIVE:
To analyze the Epstein-Barr virus (EBV) load in different lymphocyte subsets, as well as clinical characteristics and outcomes in patients with hematologic malignancies experiencing EBV reactivation.
METHODS:
Peripheral blood samples from patients were collected. B, T, and NK cells were isolated sorting with magnetic beads by flow cytometry. The EBV load in each subset was quantitated by real-time quantitative polymerase chain reaction (RT-qPCR). Clinical data were colleted from electronic medical records. Survival status was followed up through outpatient visits and telephone calls. Statistical analyses were performed using SPSS 25.0.
RESULTS:
A total of 39 patients with hematologic malignancies were included, among whom 35 patients had undergone allogeneic hematopoietic stem cell transplantation (allo-HSCT). The median time to EBV reactivation was 4.8 months (range: 1.7-57.1 months) after allo-HSCT. EBV was detected in B, T, and NK cells in 20 patients, in B and T cells in 11 patients, and only in B cells in 4 patients. In the 35 patients, the median EBV load in B cells was 2.19×104 copies/ml, significantly higher than that in T cells (4.00×103 copies/ml, P <0.01) and NK cells (2.85×102 copies/ml, P <0.01). Rituximab (RTX) was administered for 32 patients, resulting in EBV negativity in 32 patients with a median time of 8 days (range: 2-39 days). Post-treatment analysis of 13 patients showed EBV were all negative in B, T, and NK cells. In the four non-transplant patients, the median time to EBV reactivation was 35 days (range: 1-328 days) after diagnosis of the primary disease. EBV was detected in one or two subsets of B, T, or NK cells, but not simultaneously in all three subsets. These patients received a combination chemotherapy targeting at the primary disease, with 3 patients achieving EBV negativity, and the median time to be negative was 40 days (range: 13-75 days).
CONCLUSION
In hematologic malignancy patients after allo-HSCT, EBV reactivation commonly involves B, T, and NK cells, with a significantly higher viral load in B cells compared to T and NK cells. Rituximab is effective for EBV clearance. In non-transplant patients, EBV reactivation is restricted to one or two lymphocyte subsets, and clearance is slower, highlighting the need for prompt anti-tumor therapy.
Humans
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Hematologic Neoplasms/virology*
;
Herpesvirus 4, Human/physiology*
;
Epstein-Barr Virus Infections
;
Hematopoietic Stem Cell Transplantation
;
Virus Activation
;
Lymphocyte Subsets/virology*
;
Flow Cytometry
;
Killer Cells, Natural/virology*
;
Male
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Female
;
B-Lymphocytes/virology*
;
Viral Load
;
Adult
;
T-Lymphocytes/virology*
;
Middle Aged
2.Rapid characterization and identification of non-volatile constituents in Pogostemonis Herba by UPLC-Q-TOF-MS combined with UNIFI and an in-house library.
Chao-Fa WEI ; Li-Wan ZHU ; Xiao-Yan LAN ; Li ZHOU ; Xiang LI ; Qing-Xiu HAO ; Zi-Dong QIU ; Li-Ping KANG
China Journal of Chinese Materia Medica 2023;48(14):3826-3838
This study aimed to characterize and identify the non-volatile components in Pogostemonis Herba by using ultra-perfor-mance liquid chromatography-quadrupole-time of flight-mass spectrometry(UPLC-Q-TOF-MS) combined with UNIFI and an in-house library. The chemical components in 50% methanol extract of Pogostemonis Herba were detected by UPLC-Q-TOF-MS in both positive and negative MS~E continuum modes. Then, the MS data were processed in UNIFI combined with an in-house library to automatically characterize the metabolites. Based on the multiple adduct ions, exact mass, diagnostic fragment ions, and peak intensity of compounds and the fragmentation pathways and retention behaviors of reference substances, the structures identified by UNIFI were further verified and those of the unidentified compounds were tentatively elucidated. A total of 120 compound structures were identified or tentatively identified, including flavonoids, phenylpropanoids, phenolic acids, terpenes, fatty acids, alkaloids, and phenylethanoid glycosides. Sixteen of them were accurately identified by comparison with reference substances, and 53 compounds were reported the first time for Pogostemonis Herba. This study systematically characterized and identified the non-volatile compounds in Pogostemonis Herba for the first time. The findings provide a scientific basis for revealing the pharmacodynamic material basis, establishing a quality control system, and developing products of Pogostemonis Herba.
3.Prevalence and risk factors of obesity in children with Diamond-Blackfan anemia.
Mei-Hui YI ; Yang WAN ; Si-Qi CHENG ; Xiao-Wen GONG ; Zi-Xi YIN ; Jun LI ; Yang-Yang GAO ; Chao WU ; Su-Yu ZONG ; Li-Xian CHANG ; Yu-Mei CHEN ; Rong-Xiu ZHENG ; Xiao-Fan ZHU
Chinese Journal of Contemporary Pediatrics 2022;24(10):1143-1148
OBJECTIVES:
To investigate the distribution of body mass index (BMI) and risk factors for obesity in children with Diamond-Blackfan Anemia (DBA).
METHODS:
The children with DBA who attended National Clinical Research Center for Blood Diseases, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, from January 2003 to December 2020 were enrolled as subjects. The related clinical data and treatment regimens were recorded. The height and weight data measured within 1 week before or after follow-up time points were collected to calculate BMI. The risk factors for obesity were determined by multivariate regression analysis in children with DBA.
RESULTS:
A total of 129 children with DBA were enrolled, among whom there were 80 boys (62.0%) and 49 girls (38.0%), with a median age of 49 months (range 3-189 months). The prevalence rate of obesity was 14.7% (19/129). The multivariate logistic regression analysis showed that the absence of ribosomal protein gene mutation was closely associated with obesity in children with DBA (adjusted OR=3.63, 95%CI: 1.16-11.38, adjusted P=0.027). In children with glucocorticoid-dependent DBA, obesity was not associated with age of initiation of glucocorticoid therapy, duration of glucocorticoid therapy, and maintenance dose of glucocorticoids (P>0.05).
CONCLUSIONS
There is a high prevalence rate of obesity in children with DBA, and the absence of ribosomal protein gene mutation is closely associated with obesity in children with DBA.
Child
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Male
;
Female
;
Humans
;
Anemia, Diamond-Blackfan/genetics*
;
Pediatric Obesity/complications*
;
Glucocorticoids/therapeutic use*
;
Prevalence
;
Risk Factors
;
Ribosomal Proteins/genetics*
;
Mutation
4.Surveillance of epidemic factors of schistosomiasis in wetlands in Sichuan Province
Yong MAO ; Jia XU ; Liang XU ; Ling CHEN ; Jia-Jia WAN ; Rong-Zhi LI ; Meng TANG ; Chao-Kun ZHONG ; Fang YANG ; Hui-Rong XU ; Yu-Ling YE ; Zi-Song WU
Chinese Journal of Schistosomiasis Control 2019;31(3):307-310
Objective To investigate the epidemic factors of schistosomiasis in wetlands in Sichuan Province, so as to provide scientific evidence for the prevention and control of schistosomiasis in wetlands. Methods The artificial and natural wetlands were selected from Sichuan Province, and the relevant data regarding wetlands were collected. Routine snail survey, investigation on human morbidity due to schistosomiasis, snail diffusion experiments, questionnaire survey, determination of water infectivity and retrospective survey were conducted. Results In Sichuan Province, the mean densities of living snails were 0.003 snails/0.1 m2 and 2.033 snails/0.1 m2 in the upper and lower reaches of the Meiwan Reservoir wetlands, 0.08 snails/0.1 m2 in the Jinyan Lake of Guanghan City, 0.21 snails/0.1 m2 in Muhe River of Guanghan City, and 0.02 snails/0.1 m2 prior to the construction of Qiong-hai wetland park in Xichang City in 2015. Artificial simulation experiments showed that the largest distance of snail diffusion in water was 2 000 m. There were 8.80% (41/466) of subjects that lived neighboring wetlands, worked in wetlands and visited wet-lands having infested water contact behaviors. A total of 690 sentinel mice were assigned, and no Schistosoma japonicum infection was detected in the 677 mice dissected. Retrospective survey showed that the construction of the Meiwan Reservoir caused the spread of schistosomiasis in Dailing County, and snails were found in the ditches entering the Jinyan Lake and in Jinyan Lake areas 5 years following the construction of the Jinyan Lake in Guanghan City, with S. japonicum -infected snails detected in the ditches entering the Jinyan Lake. Conclusions O. hupensis snails are found in some wetlands in Sichuan Province. Protection of wetlands and snail control with environmental improvements are recommended for the prevention of snail importation in natural wetlands, while in artificial wetlands, thorough snail control is recommended during the construction of the wetlands because of the likelihood of snail importation via water systems. In addition, both natural and artificial wetlands require long-term systematic surveillance of schistosomiasis.
5.Roles of Loss of Chromosome 14q Allele in the Prognosis of Renal Cell Carcinoma with C-reactive Protein Abnormity.
Gang WANG ; Da-Ming ZHANG ; Hai-Ying ZHUANG ; Chao YIN ; Jing LIU ; Zi-Chun WANG ; Li-Cheng CAI ; Ming-Hua REN ; Wan-Hai XU ; Cheng ZHANG
Chinese Medical Journal 2017;130(18):2176-2182
Background:Renal cell carcinoma (RCC) is frequently associated with paraneoplastic inflammatory syndrome (PIS). This study aimed at exploring the connections between the survival rate and specific gene alterations and the potential mechanism.
Methods:We retrospectively studied 69 surgical RCC cases from August 2014 to February 2016, including 18 cases of clear cell RCC (ccRCC) demonstrating elevated pretreatment serum C-reactive protein (CRP, Group A). Twelve of the 18 cases were symptomized with febrile episode. We also selected 49 cases of ccRCC with normal pretreatment CRP (Group B). Using 22 microsatellite markers, we compared the incidence of loss of heterozygosity (LOH) between Group A and Group B. All statistical tests are two-sided.
Results:The 3p LOH was common in both Group A (89%) and Group B (92%). The frequency of 14q LOH in Group A (16 of 18) was higher than Group B (4 of 49, χ= 40.97 P < 0.0001). The 3p and 14q LOH were the characteristics of ccRCC with elevated acute phase reactants, including PIS, regardless of the presence of metastasis. On the contrary, 14q LOH was a rare genomic alternation in advanced-staged ccRCC without PIS. The overall survival of patients with elevated CRP (33.3%) was lower than its counterparts (6.1%, hazard ratio=1.852, P < 0.0001) in Kaplan-Meier curve.
Conclusions:The results imply that the disruption of a 14q gene(s) might result in not only the inflammatory manifestations in the tumor host but also the poor survival rate as well. The isolation of the gene(s) on 14q might be a vital goal in the treatment of PIS-associated RCC.
7.Construction and identification of lentiviral vector for RNA interference targeting STUB1 gene.
Hong ZHAO ; Jing-yu ZHANG ; Wan-hai XU ; Zi-chao YANG ; Qing-jie ZHAO
Journal of Zhejiang University. Medical sciences 2010;39(6):623-627
OBJECTIVETo construct and identification of a lentiviral vector for RNA interference (RNAi) targeting STUB1 gene.
METHODSA pair of complementary small hairpin RNA (shRNA) oligonucleotides targeting STUB1 gene was designed, synthesized and inserted into linearized pMagic 4.0 vector. The recombinant plasmid was identified by double restriction digestion with Age I/EcoR I and DNA sequencing.
RESULTPCR and DNA sequencing showed that the shRNA sequence was successfully inserted into pMagic 4.0 vector. The pMagic 4.0 vector was successfully packaged into lentivirus particles.
CONCLUSIONA lentiviral shRNA expression vector and particles targeting STUB1 gene has been successfully constructed for the further study of the STUB1 gene.
Gene Targeting ; Genetic Vectors ; Lentivirus ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Ubiquitin-Protein Ligases ; genetics
8.Detection and clinical significance of JAK2 mutation in 412 patients with chronic myeloproliferative neoplasms.
Hong-Ying CHAO ; Zheng FAN ; Ri ZHANG ; Yi-Min SHEN ; Wan CHEN ; Hai-Rong FEI ; Zi-Ling ZHU ; Yu-Feng FENG ; Zi-Xing CHEN ; Yong-Quan XUE
Chinese Journal of Oncology 2009;31(7):510-514
OBJECTIVETo investigate the frequency of JAK2V617F mutation in Chinese patients with chronic myeloproliferative neoplasms (MPN) and to study the relationship between JAK2V617F mutation and clinical characteristics.
METHODSJAK2V617F mutation was screened by allele-specific polymerase chain reaction (AS-PCR).
RESULTSJAK2V617F mutation was detected in 277 of the 412 patients with MPN. The frequency of JAK2V617F mutation was similar among essential thrombocythemia (ET), idiopathic myelofibrosis (IMF) and chronic myeloproliferative disorders-unclassified (MPD-U) (P > 0.05), but it was significantly lower than that in polycythemia vera (PV) (P < 0.05). The presence of JAK2V617F was found to be significantly correlative with advanced age at diagnosis (P < 0.01) and with higher hemoglobin levels and higher leukocyte counts (P < 0.05). Significant difference was found in complication of vascular events between JAK2V617 positive and negative patients (P < 0.05). JAK2V617F positive MPD-U patients were more prone to progress into typical MPN compared with JAK2V617F negative MPD-U patients. The association between abnormal karyotype and JAK2V617F was not found in cytogenetical analysis of 301 patients.
CONCLUSIONThe presence of JAK2V617F in MPD-U is associated with the disease development. There is a correlation between JAK2V617F mutation in MPN and advanced age, higher leukocyte counts, hemoglobin level and vascular events.
Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Hemoglobins ; metabolism ; Humans ; Janus Kinase 2 ; genetics ; Leukocyte Count ; Male ; Middle Aged ; Mutation ; Myeloproliferative Disorders ; blood ; complications ; genetics ; Polycythemia Vera ; blood ; complications ; genetics ; Primary Myelofibrosis ; blood ; complications ; genetics ; Thrombocythemia, Essential ; blood ; complications ; genetics ; Thrombosis ; etiology ; Young Adult
9.Micro-CT evaluation and histological analysis of screw-bone interface of expansive pedicle screw in osteoporotic sheep.
Shi-yong WAN ; Wei LEI ; Zi-xiang WU ; Rong LV ; Jun WANG ; Suo-chao FU ; Bo LI ; Ce ZHAN
Chinese Journal of Traumatology 2008;11(2):72-77
OBJECTIVETo investigate the properties of screw-bone interface of expansive pedicle screw (EPS) in osteoporotic sheep by micro-CT and histological observation.
METHODSSix female sheep with bilateral ovariectomy-induced osteoporosis were employed in this experiment. After EPS insertion in each femoral condyle, the sheep were randomly divided into two groups: 3 sheep were bred for 3 months (Group A), while the other 3 were bred for 6 months (Group B). After the animals being killed, the femoral condyles with EPS were obtained, which were three-dimensionally-imaged and reconstructed by micro-CT. Histological evaluation was made thereafter.
RESULTSThe trabecular microstructure was denser at the screw-bone interface than in the distant parts in expansive section, especially within the spiral marking. In the non-expansive section, however, there was no significant difference between the interface and the distant parts. The regions of interest (ROI) adjacent to EPS were reconstructed and analyzed by micro-CT with the same thresholds. The three-dimensional (3-D) parameters, including tissue mineral density (TMD), bone volume fraction (BVF, BV/TV), bone surface/bone volume (BS/BV) ratio, trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp), were significantly better in expansive sections than non-expansive sections (P less than 0.05). Histologically, newly-formed bony trabeculae crawled along the expansive fissures and into the center of EPS. The newly-formed bones, as well as the bones at the bone-screw interface, closely contacted with the EPS and constructed four compartments.
CONCLUSIONSThe findings of the current study, based on micro-CT and histological evaluation, suggest that EPS can significantly provide stabilization in osteoporotic cancellous bones.
Animals ; Bone Screws ; Bone and Bones ; diagnostic imaging ; pathology ; Female ; Osteoporosis ; diagnostic imaging ; pathology ; Sheep ; Tomography, X-Ray Computed
10.Micro-CT evaluation and histological analysis of screw-bone interface of expansile pedicle screw in osteoporotic sheep.
Shi-yong WAN ; Wei LEI ; Zi-xiang WU ; Rong LÜ ; Jun WANG ; Bo LI ; Suo-Chao FU ; Ce ZHAN
Chinese Journal of Surgery 2007;45(18):1271-1273
OBJECTIVETo investigate the properties of screw-bone interface of expansive pedicle screw (EPS) in osteoporotic sheep using micro-CT and histology.
METHODSSix female sheep with bilateral ovariectomy-induced osteoporosis were employed in this experiment and were randomly assigned into 2 groups: A and B. After EPS insertion in each femoral condyles, sheep in group A were bred for 3 months, while those in group B 6 months. Femoral condyles with EPS were 3D-imaged and reconstructed by micro-CT. Histology was evaluated thereafter.
RESULTSThe trabecular microstructure was denser at the screw-bone interface than in the distant parts in expansive section, especially within spiral marking. In the non-expansive section, however, there was no significant difference between the interface and the distant parts. The regions of interest (ROI) adjacent to EPS were reconstructed and analyzed by micro-CT using the same thresholds. The 3D-parameters generated, including tissue mineral density (TMD), bone volume fraction (BVF, BV/TV), bone surface/bone volume (BS/BV) ratio, trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp), were significantly better in expansive than in non-expansive sections (P < 0.05). Histologically, newly formed bone trabeculae crawled along the expansive fissures and into the center of EPS. The newly formed bones, as well as the bone at the bone-screw interface, closely contacted the EPS and constructed four compartments.
CONCLUSIONSBased on micro-CT and histological evaluation, the study suggest that EPS can significantly provide stabilization in osteoporotic cancellous bone.
Animals ; Bone Density ; Bone Screws ; Disease Models, Animal ; Female ; Femur ; diagnostic imaging ; metabolism ; surgery ; Osteoporosis ; etiology ; pathology ; surgery ; Ovariectomy ; adverse effects ; Reproducibility of Results ; Sheep ; Tomography, X-Ray Computed ; methods

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