1.Bioinformatic analysis of the sequences of lamins from different species
Henglu ZHANG ; Huimin HUANG ; Zhixu YE ; Zhihua WANG ; Zhixu HE ; Liping SHU
Acta Laboratorium Animalis Scientia Sinica 2016;24(4):351-357
Objective Type A lamins are encoded by LMNA and a major component of the nuclear lamina, which have been suggested to play important roles in chromatin organization, transcription, DNA replication, and cell apoptosis. The aim of this study was to analyze the bioinformation of zebrafish lamins. Methods A phylogeny analysis was figured out with protein sequences of different species by Clustal X and MEGA 4?0 software. Then we compared the lamin protein sequences of different species with that of zebrafish by BLAST tool from NCBI. A figure of synteny analysis results was done with lamin sequence information of humans, murine and zebrafish cited from UCSC, Vega and Ensemble. Results The a?nalysis results showed that lmna, lmnb1, and lmnb2 genes of zebrafish are highly conservative and they may be homology of human LMNA, LMNB1 and LMNB2 genes. Conclusions Zebrafish lamins and human lamins have homologous sequence similarity, indicating that these two genes are orthologous genes.
2.In vitro hematopoietic stem cells from embryonic stem cells reconstruct hematopoiesis in mice
Zhixu HE ; Bing LIN ; Shaoliang HUANG ; Qiang MI ; Jing HUANG
Chinese Journal of Pathophysiology 2000;0(08):-
AIM:To investigate the potential of hematopoietic stem cells(HSCs)derived from mouse embryonic stem cells(ESCs)to reconstruct hematopoiesis in vivo.METHODS:Using a three-step method,a mice embryonic stem cell line,E14.1 was induced into hematopoietic stem cells.The cell markers with CD34+/Sca-1+ were identified by flow cytometry analysis,then HSCs(1?109 cells/L)from third-step were injected into SCID mice for observing teratoma formation.To validate function of HSCs,colonogenic cell assay was conducted and the hematopoiesis in lethally irradiated mice was reconstituted.RESULTS:The method of three-step differentiation,combined to use more hematopoietic stimulating factor promoted the E14.1 cell differentiation into HSCs with highest percent of CD34+/Sca-1+ cells(as high as 58.64%?4.20%)with more CFU-E,CFU-GM and CFU-GEMM populations.The cells showed the character of hematopoietic progenitors by Wright-Giemsa staining.Positive selected CD34+/Sca-1+ cells by magnetic sorting from third-step differentiation were transplanted into 7 lethally irradiated female mice while predominant hematopoietic reconstitution were observed in 10 d after transplantation,with 71.4%(5/7)successful engraftment rate.Three recipients showed that the cell population of the peripheral blood leukocytes,red blood cells and hemoglobin approached to normal index at 40 d after transplantation,but followed relative slow renew in platelet count.Survival rate in transplant group was 43%,compared to 100% mortality in control mice.Karyotyping assays confirmed the female mice with XY.CONCLUSION:The three-step differentiation and the culture conditions described here support the differentiation of mouse ESCs into HSCs.HSCs derived from mouse ESCs can reconstruct hematopoiesis.
3.Progress in reIations of zinc compounds with Zn(N2O2)coordination mode and L-theanine-zinc with diabetes meIIitus
Honggui WAN ; Yincheng PENG ; Zhixu HUANG ; Xiangju YAN
Chinese Journal of Pharmacology and Toxicology 2014;(5):774-778
Zinc and its compounds with insulin-like effects have been found to own such properties as strong bio-compatibility,good stability and remarkable hypoglycemic effect. Since the discovery of their insulin-like function,zinc compounds and their anti-diabetic effects have been a geobal focus of research. Zinc ion is involved in physiological and biochemical processes in vivo,such as in carbohy-drate and lipid metabolism. The mechanism of zinc and zinc complexes in anti-diabetic activity are sum-marized in this article,including direct participation in insulin synthesis,antioxidation,involvement in the regulation of tyrosine protein phosphatase,and improving immunity. Zinc complexes with Zn(N2 O2 )have remarkable insulin-like effects. Particularly,the L-theanine-zinc compound,which is a class of zinc com-plexes with natural ligands,has also been found to show significant hypoglycemic effect in an in vivo experiment on KK-Ay mice,have stable chemical properties,be absorbed fast in vivoand be non-toxic with oral administration during model animal experiments. This paper reviews the progress in research on relationships between zinc and its complexes and the anti-diabetic effect.
4.Primary study on inductive methods of hematopoietic stem cellfrom embryonic stem cells in vitro
Zhixu HE ; Shaoliang HUANG ; Qifeng ZHOU ; Shunong LI
Chinese Journal of Pathophysiology 1986;0(04):-
AIM: To explore inductive methods of hematopoietic stem cell (HSC) from embryonic stem cells (ESC) in vitro. METHODS: Using mice E14 line, the first step was the primary differentiation in which the ESC form embryoid bodies (EB) in methylcellulose-based cultures with SCF and VEGF. The second step involved the plating of cells originating from the EB into three different system of cultures containing SCF, VEGF,IL-3, IL-6 and EPO for HSC. And identifying HSC by flow cytometry analysis, colonogenic cells assay and Wright-Giemsa stain were also used. RESULTS: By two-step differentiating, it showed that HSC differentiated slowly in methylcellulose medium, percent age of CD34+/Sca-1+ cells slight increased about(31.5?4.7)% after day 14 induction. However, EBs were induced after 10 days to fast differentiate for HSC with more cells population by coculture on bone marrow stromal cells feeder. Flow cytometry analysis showed that percentages of CD34+/Sca-1+ cells might reached to (47.8?6.3)%. The more optimistic system of differentiation was bone marrow stromal cells feeder (BMSCF) in combination with supernatants of stromal cells from mice fetal liver(SSCFL), it significant supported differentiation of ESC into HSC with higher percent (53.6?7.2)%. Colonogenic cell assay and Wright-Giemsa stain confirmed that it possessed character of hematopoietic progenitors. CONCLUSION: Using methods of two-step differentiation, mice ESC were induced to differentiate into HSC by coculturing with BMSCF and SSCFL in combination of SCF,VEGF,IL-3,IL-6 and EPO.
5.Study on the lmna gene knockdown in zebrafish embryo with morpholino oligonucleotides
Feng LIU ; Huimin HUANG ; Zhihua WANG ; Xijun WU ; Zhixu HE ; Liping SHU
Chinese Journal of Comparative Medicine 2016;26(8):85-90
Objective Lamins are the major components of nuclear lamina underneath the inner nuclear membrane (INM).Lamins express in most cells and are involved in the whole process of growth, also play a major role in cell stability and embryonic development.Mutant in human LMNA gene may lead to a series of disorders, which are similar to progeria or other aging-associate syndrome.In this study, we report a new lmna knockdown animal model generated in our laboratory in order to provide a useful tool for studying laminopathies.Methods Two plasmids tagged to zebrafish lmna gene were designed based on morpholino oligonucleotides technology.Co-microinjected the plasmids into zebrafish embryos to knockdown lmna gene.Imagining and western blot detection were used to identify the mutants.Results Two different proteins, Lamin A/C, were expressed in the zebrafish embryos.Two plasmids lmna-MO and lmna-EGFP-pCS 2 + were generated and co-microinjected into embryos.The results of imagining and western blot showed that the expression of lmna gene was downregulated in the zebrafish embryos.Conclusions Lamin A/C are expressed in zebrafish.lmna gene can be knocked down by the injection of lmna-MO and lmna-EGFP-pCS 2 +.This new animal model may be a powerful tool for study on laminopathies.
6.Spatiotemporal expression spectrum of Rap1 gene in zebrafish early development process
Xiaoyan YANG ; Zhixu HE ; Liping SHU ; Jiao JIN ; Jing HUANG ; Sasa WU ; Jianjuan MA
Chongqing Medicine 2016;45(20):2748-2751
Objective To choose zebrafish as the experimental animal model for studying the spatiotemporal expression rule of rap1 gen in zebrafish embryo early development process .Methods The Rap1 gene fragment was cloned from the zebrafish emby‐oscDNA ,then the Rap1 gene fragment and pCS2+ plasmid were performed the in vitro connetion and recombination was extracted , the combinant plasmid was correct after the double enzyme digestion ,colony PCR and sequencing identification .T3 RNA polymer‐ase in vitro transcription system was used to obtain the digoxin (DIG)‐labeled anti‐sense mRNA probe of Rap1 gene .The whole mount in situ hybridization method was adopted to detect the Rap1 expression in zebrafish embryo early development process . Results The positive hybridization signal of Rap1 gene was detected at the cell division junction region of 0 .75 hpf ,animal pole of 3 .70 hpf and 6 .00 hpf ,and notochord of 12 .00 -72 .00 hpf .Conclusion Rap1 gene might be involved in the early development process of notochord nervous system in zebrafish .
7. Prognostic factors in immune thrombocytopenia in children
Ruyan TONG ; Jiao JIN ; Jing HUANG ; Guifang CHEN ; Zhixu HE
Chinese Journal of Applied Clinical Pediatrics 2019;34(11):837-841
Objective:
To explore the prognostic relationship between initial absolute lymphocyte count(ALC) of peripheral blood and primary immune thrombocytopenia (ITP) in children, in order to provide basis for judging the prognosis and treatment of ITP in children.
Methods:
Clinical data of 166 children with primary ITP in children admi-tted at the Affiliated Hospital of Guizhou Medical University from January 2014 to March 2018 were analyzed retrospectively, and they were followed up by clinic and telephone, the prognostic factors (gender, age, ethnicity, inducement, bleeding, initial ALC, platelet count and treatment) were statistically analyzed, and the relationship between ALC of peripheral blood and the prognosis of children with ITP was observed.
Results:
Of 166 children with ITP, 89 cases (53.6%) had remission within 3 months, 18 cases (10.8%) within 3-12 months, 20 cases (12.0%) within 1-4 years, a total of 39 cases (23.5%) were refractory (no remission in 1-10 years), the remission rate within 1 year was 64.5%, the total remission rate was 76.4%.ALC of remission cases was (4.58±2.87)×109/L within 3 months, (4.47±2.04)×109/L within 3-12 months, and (2.86±1.61)×109/L within 1- 4 years.Thirty-nine cases (23.5%) were refractory(no remission in 1-10 years), ALC of them was (2.07±0.98)×109/L, and there were significant differences among different groups (
8.Evaluation value of IL-6,PCT,IgG in children with hormone-sensitive PNS bacteria infection pneumonia susceptibility and recurrence
Chunling MA ; Zhong HUANG ; Jing ZHOU ; Xin XIE ; Zhixu YE
International Journal of Laboratory Medicine 2024;45(17):2103-2107
Objective To investigate the value of interleukin-6(IL-6),procalcitonin(PCT)and immuno-globulin G(IgG)in evaluating the susceptibility and recurrence of bacterial pneumonia in children with hor-mone-sensitive primary nephrotic syndrome(PNS).Methods A total of 150 children with hormone-sensitive PNS admitted to the hospital from January 2019 to March 2023 were selected as the study objects.The pa-tients were divided into concurrent group and non-concurrent group according to whether bacterial pneumonia was complicated or not within 3 months.The baseline IL-6,PCT and IgG levels of the two groups were com-pared,and the factors affecting susceptibility to bacterial pneumonia were analyzed by Logistic regression.Re-ceiver operating characteristic curve(ROC)was drawn to analyze the value of IL-6,PCT and IgG in assessing susceptibility to PNS bacterial pneumonia and the risk of PNS recurrence.Results IL-6[(7.84±2.49)pg/mL]and PCT[(0.39±0.11)ng/mL]in concurrent group were higher than those in non-concurrent group[(4.75±1.56)pg/mL,(0.29±0.09)ng/mL].IgG[(3.82±1.25)g/L]was lower than that of non-concurrent group[(5.92±1.97)g/L],and the differences were statistically significant(P<0.05).Logistic regression e-quation showed that with the increase of IL-6 and PCT and the decrease of IgG,the susceptibility to bacterial pneumonia gradually increased,and the difference was statistically significant(P<0.05).The AUC of IL-6,PCT and IgG for PNS bacterial pneumonia susceptibility was 0.792,0.744 and 0.677,respectively.The AUC of IL-6 and PCT combined IgG for PNS bacterial pneumonia susceptibility was 0.929,which was higher than that of IL-6,PCT and IgG alone(P<0.05).The risk of PNS recurrence in patients with high IL-6 level was 7.054 times that of low IL-6 level,the risk of PNS recurrence in patients with high PCT level was 6.750 times that of low PCT level,and the risk of PNS recurrence in patients with high IgG level was 0.079 times that of low level,the differences were statistically significant(P<0.05).Conclusion IL-6,PCT and IgG are the influencing factors of hormone-sensitive PNS bacterial pneumonia in children,and they are related to the recurrence of PNS.Clinically,IL-6,PCT and IgG levels can be used to evaluate the susceptibility and recur-rence risk of bacterial pneumonia in children,so as to provide guidance for preventive clinical intervention.
9.Analysis of malnutrition status in pediatric acute lymphoblastic leukemia patients dur-ing the induced remission stage
Jing HUANG ; Jing LI ; Zhixu HE ; Jiao JIN ; Shasha WU ; Huiqun WANG
Chinese Journal of Clinical Oncology 2018;45(21):1096-1099
Objective: To investigate the nutritional status of children acute lymphoblastic leukemia (ALL) during induced remission stage. The effects of the disease itself, treatment, and complications of malnutrition were all analyzed. Methods: From the medical re-cords of children with ALL in the pediatric hematological department of the Affiliated Hospital of Guizhou Medical University, we col-lected basic information of the children; monitored the height and weight of the children on the first, 15th, and 33rd days of induced remission treatment; and calculated their body mass index (BMI), as well as results of laboratory tests such as serum albumin and se-rum prealbumin. SPSS 23.0 software was used for data analysis. Results: In 40 children with ALL, there were 16 cases of malnutrition on the first day of induced remission treatment accounting for 40%, 14 cases on both the 15th day and the 33rd day of treatment ac-counting for 35%. Among children with malnutrition, 4 cases of moderate to severe malnutrition occurred on the first day of treat-ment accounting for 25.0%, 9 cases on the 15th day accounting for 64.3%, and 12 cases on the 33rd day accounting for 85.7%. Com-pared with day 1 and day 33, the difference was statistically significant (P<0.017). In the induced remission period, the BMI on the first day was (15.98±2.17) kg/m2; on the 15th day, it was (15.65±2.20) kg/m2; and on the 33rd day, it was (15.66±1.92) kg/m2. The differ-ence between the three was not statistically significant (P=0.730). In the single-factor analysis of factors related to the nutritional sta-tus of children, infection, digestive system involvement, and serum albumin levels were related to the occurrence of malnutrition, and we performed multifactor analysis of these three factors. The difference between the level of infection and serum albumin and the oc-currence of malnutrition was statistically significant (P<0.05). Conclusions: During the induced remission, the malnutrition degree of some children with ALL was aggravated. Infection was a high-risk factor for malnutrition in children with ALL. The decrease in serum al-bumin level may indicate the occurrence of malnutrition. Dynamic monitoring of nutritional status in children with ALL is necessary.
10.Background, design, and preliminary implementation of China prospective multicenter birth cohort
Si ZHOU ; Liping GUAN ; Hanbo ZHANG ; Wenzhi YANG ; Qiaoling GENG ; Niya ZHOU ; Wenrui ZHAO ; Jia LI ; Zhiguang ZHAO ; Xi PU ; Dan ZHENG ; Hua JIN ; Fei HOU ; Jie GAO ; Wendi WANG ; Xiaohua WANG ; Aiju LIU ; Luming SUN ; Jing YI ; Zhang MAO ; Zhixu QIU ; Shuzhen WU ; Dongqun HUANG ; Xiaohang CHEN ; Fengxiang WEI ; Lianshuai ZHENG ; Xiao YANG ; Jianguo ZHANG ; Zhongjun LI ; Qingsong LIU ; Leilei WANG ; Lijian ZHAO ; Hongbo QI
Chinese Journal of Perinatal Medicine 2024;27(9):750-755
China prospective multicenter birth cohort (Prospective Omics Health Atlas birth cohort, POHA birth cohort) study was officially launched in 2022. This study, in collaboration with 12 participating units, aims to establish a high-quality, multidimensional cohort comprising 20 000 naturally conceived families and assisted reproductive families. The study involves long-term follow-up of parents and offspring, with corresponding biological samples collected at key time points. Through multi-omics testing and analysis, the study aims to conduct multi-omics big data research across the entire maternal and infant life cycle. The goal is to identify new biomarkers for maternal and infant diseases and provide scientific evidence for risk prediction related to maternal diseases and neonatal health.