Clinical characteristics of intestinal stricture after neonatal necrotizing enterocolitis / 国际儿科学杂志

Neonatal necrotizing enterocolitis(NEC)is a common acute abdomen in newborns, while intestinal stricture is one of the frequent complications of NEC.Post-NEC stricture often occurs in the colon, and has clinical features such as vomiting, abdominal distension and bloody stools.This complication has a high incidence, high risk of death, and is also affected by multiple factors such as disease severity, treatment method and recovery time of enteral nutrition.Early prediction, diagnosis and intervention can reduce the adverse effects of the disease on the growth and development of children.This article reviews the clinical characteristics, influencing factors and prediction of the post-NEC stricture.

VPS4B mutation impairs the osteogenic differentiation of dental follicle cells derived from a patient with dentin dysplasia type I / 国际口腔科学杂志·英文版

A splicing mutation in VPS4B can cause dentin dysplasia type I (DD-I), a hereditary autosomal-dominant disorder characterized by rootless teeth, the etiology of which is genetically heterogeneous. In our study, dental follicle cells (DFCs) were isolated and cultured from a patient with DD-I and compared with those from an age-matched, healthy control. In a previous study, this DD-I patient was confirmed to have a loss-of-function splicing mutation in VPS4B (IVS7 + 46C > G). The results from this study showed that the isolated DFCs were vimentin-positive and CK14-negative, indicating that the isolated cells were derived from the mesenchyme. DFCs harboring the VPS4B mutation had a significantly higher proliferation rate from day 3 to day 8 than control DFCs, indicating that VPS4B is involved in cell proliferation. The cells were then replenished with osteogenic medium to investigate how the VPS4B mutation affected osteogenic differentiation. Induction of osteogenesis, detected by alizarin red and alkaline phosphatase staining in vitro, was decreased in the DFCs from the DD-I patient compared to the control DFCs. Furthermore, we also found that the VPS4B mutation in the DD-I patient downregulated the expression of osteoblast-related genes, such as ALP, BSP, OCN, RUNX2, and their encoded proteins. These outcomes confirmed that the DD-I-associated VPS4B mutation could decrease the capacity of DFCs to differentiate during the mineralization process and may also impair physiological root formation and bone remodeling. This might provide valuable insights and implications for exploring the pathological mechanisms underlying DD-I root development.

Structural characteristics of the deciduous teeth of Tibetan miniature pigs / 南方医科大学学报

OBJECTIVE@#To explore the histological structure of the deciduous teeth and the tooth germs of Tibetan miniature pigs for studies of dental tissue diseases and tooth regeneration.@*METHODS@#The structure of the deciduous teeth of Tibetan miniature pigs was observed by X-ray. The ultrastructure of the enamel and dentin of deciduous teeth was characterized by scanning electron microscopy. The jaws and teeth were three-dimensionally reconstructed using Mimics software based on Micro-CT scanning of the deciduous teeth. Image J software was used to calculate the gray value and the mineralization density of the deciduous teeth. Hisotological structure of the tooth germ and the pulp tissue of Tibetan miniature pigs was observed using HE staining.@*RESULTS@#The deciduous teeth of Tibetan miniature pigs were composed of enamel, dentin and medullary pulp tissue. The permanent tooth germ were formed during the deciduous dentition. The enamel and dentin ultrastructure of deciduous teeth were consistent with that of human deciduous teeth. The enamel and dentin mineralization densities were 2.47±0.09 g/cm and 1.72±0.07 g/cm, respectively. The pathological structures of tooth germ and pulp tissue were similar to those of human teeth, and the pulp tissue of the deciduous teeth was in an undifferentiated state.@*CONCLUSIONS@#The deciduous teeth of Tibetan miniature pig have similar anatomy, ultrastructure and histopathological structure to human teeth and can serve as a good animal model for studying human dental tissue diseases and the mechanisms of tooth regeneration.

Progress of cholestasis liver diseases with low serum γ-glutamyl transpeptidase level / 中华实用儿科临床杂志

γ-glutamyl transpeptidase(GGT) is an important index of cholestasis.Most patients with cholestasis liver diseases have high GGT level, but some of the patients are characterized by low GGT level.This paper summarizes the progress of cholestasis liver diseases with low GGT level in order to improve the understanding of these diseases.

Progress of cholestasis liver diseases with low serum γ-glutamyl transpeptidase level / 中华实用儿科临床杂志

γ-glutamyl transpeptidase (GGT) is an important index of cholestasis.Most patients with cholestasis liver diseases have high GGT level,but some of the patients are characterized by low GGT level.This paper summarizes the progress of cholestasis liver diseases with low GGT level in order to improve the understanding of these diseases.

Whole exome sequencing identifies an AMBN missense mutation causing severe autosomal-dominant amelogenesis imperfecta and dentin disorders / 国际口腔科学杂志·英文版

Tooth development is a complex process that involves precise and time-dependent orchestration of multiple genetic, molecular, and cellular interactions. Ameloblastin (AMBN, also named "amelin" or "sheathlin") is the second most abundant enamel matrix protein known to have a key role in amelogenesis. Amelogenesis imperfecta (AI [MIM: 104500]) refers to a genetically and phenotypically heterogeneous group of conditions characterized by inherited developmental enamel defects. The hereditary dentin disorders comprise a variety of autosomal-dominant genetic symptoms characterized by abnormal dentin structure affecting either the primary or both the primary and secondary teeth. The vital role of Ambn in amelogenesis has been confirmed experimentally using mouse models. Only two cases have been reported of mutations of AMBN associated with non-syndromic human AI. However, no AMBN missense mutations have been reported to be associated with both human AI and dentin disorders. We recruited one kindred with autosomal-dominant amelogenesis imperfecta (ADAI) and dentinogenesis imperfecta/dysplasia characterized by generalized severe enamel and dentin defects. Whole exome sequencing of the proband identified a novel heterozygous C-T point mutation at nucleotide position 1069 of the AMBN gene, causing a Pro to Ser mutation at the conserved amino acid position 357 of the protein. Exfoliated third molar teeth from the affected family members were found to have enamel and dentin of lower mineral density than control teeth, with thinner and easily fractured enamel, short and thick roots, and pulp obliteration. This study demonstrates, for the first time, that an AMBN missense mutation causes non-syndromic human AI and dentin disorders.

Analysis of DSPP gene mutation in a Chinese pedigree affected with hereditary dentinogenesis imperfecta / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the clinical phenotype of a Chinese pedigree affected with hereditary dentinogenesis imperfecta and mutation of dentin sialophosphoprotein (DSPP) gene.</p><p><b>METHODS</b>Affected members underwent intraoral photography, dental film and panoramic radiography. Genomic DNA was extracted from peripheral venous blood samples. Coding regions of the DSPP gene were subjected to PCR amplification and Sanger sequencing. Functional effect of the mutation was predicted with SIFT and PolyPhen-2. The tertiary structure of wild type and mutant proteins were predicted by Swiss-Port.</p><p><b>RESULTS</b>A heterozygous c.50C to T (p.P17L) mutation was identified in exon 2 of the DSPP gene in the proband and her father. The same mutation was not found among 200 unrelated healthy controls. The Pro-17 residues and its surrounding positions in DSPP are highly conserved across various species. The mutation was predicted to be damaging to the structure of DSPP protein.</p><p><b>CONCLUSION</b>The c.50C to T (p.P17L) mutation of the DSPP gene probably underlies the disease in this pedigree. Above finding has expanded the spectrum of DSPP gene mutations and provided a basis for genetic counseling and prenatal diagnosis for this family.</p>

Analysis of hematological characteristics of patient with different genotypes in lightβ-thalassemia / 实用医学杂志

Objective To investigate the hematological characteristics of patients with light β- thalassae-mia and rapidly identify different mutational genotypes. Methods RBC、Hb、MCV、MCH、MCHC、RDW-CV and HbA2 were studied in the 646 patients,the differences between β0/βN and β +/βN mutations were also compared. Results Most of them were microcytic hypochromic anemia. The most common genotype were β654/βN(33%)、β41-42/βN(32.5%)、β17/βN(14.4%)、β - 28/βN(10%)respectively,β0/βN were relatively higher. The differences in RBC、MCV、MCH、RDW-CV and HbA2 were significant between β0/βN and β +/βN. Compared with β +/βN patients,the MCV and MCH of β0/βN were significantly reduced,RDW-CV and HbA2 were significantly higher. Conclusion Light β- thalassaemia with different genotypes has its own unique hematological features and can be quickly and ef-fectively identified. Clinical efficiency can be improved through hematological analysis.

Screening of CTSC gene mutations in a Chinese pedigree affected with Papillon-Lefevre syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the clinical phenotype of a Chinese pedigree affected with Papillon-Lefevre syndrome(PLS) and detect mutation of CTSC gene.</p><p><b>METHODS</b>Clinical phenotypes were noted, and oral examination for the proband was carried out for the clinical diagnosis of PLS. PCR and Sanger sequencing were used to identify potential mutation of the CTSC gene. Functional effect of the mutation was predicted with SIFT and PolyPhen-2. Swiss-Port was used to predict the tertiary structure of wild type and mutant proteins. The mRNA and protein expression were analyzed by real-time PCR and Western blotting.</p><p><b>RESULTS</b>A homozygous mutation c.901G>A (p.G301S) in exon 7 of CTSC gene was identified in the patient. Both parents of the patient had carried a heterozygous c.901G>A mutation. The mutation was located in the conserved region of CTSC enzyme and was predicted to be damaging by changing the structure of the protein, which could affect the activity of Cathepsin C. However, no significant difference was found in the expression of p.G301S variant at the mRNA and protein levels compared with that of the wild type CTSC gene.</p><p><b>CONCLUSION</b>The c.901G>A mutation of the CTSC gene was first reported in China, which has expanded its mutation spectrum.</p>

Roles of Tumor Infiltrating Lymphocytes in Hematologic Malignancies-Review / 中国实验血液学杂志

Hematologic malignancies are the most common childhood malignancy and the main cause of death in pediatric cancer patients. Systemic diagnosis and risk-based treatment in these 30 years have achived a great improvement in pediatric cancer therapy, for example, the event-free 5-years survival rate of childhood acute lymphoblastic leukemia was over 80%. Unfortunately, 20%-25% patients relapsed after-therapy, therefore, the novel therapeutic strategies that can improve survival are urgently required. T lymphocytes are the most potent effective anti-tumor cells. Tumor infiltrating lymphocytes comprise many heterotypic lymphocytes, which mainly are T cells for the adoptive cellular immunotherapy. The success of hematopoietic stem cell transplantation in leukemia therapy indicated that the functional recovery of immune cells plays a significant role in the treatment of hematologic malignancies. The cancer immunotherapy become a new strategy for malignancies, while the role of immunotherapy in hematologic malignancies still remained rarely understood. Herein, the recent research progress about the roles of tumor infiltrating lymphocyte and its potential application for hematologic malignancies are systemically reviewed.

Expression of CD163 in children with Epstein-Barr virus infection / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the clinical significance of CD163 in the diagnosis and the evaluation of severity and prognosis of childhood hemophagocytic lymphohistiocytosis (HLH).</p><p><b>METHODS</b>Ninety-four children were classified into Epstein-Barr virus (EBV)-positive (n=55) and EBV-negative groups (n=39; control group). The EBV-positive group was subgrouped into infectious mononucleosis (IM; n=47) and HLH (n=8). Serum levels of soluble CD163 were measured using ELISA. Expression of CD163 on mononuclear cells was detected by flow cytometry.</p><p><b>RESULTS</b>The serum levels of soluble CD163 were>10 000 ng/mL in all eight HLH patients (>30 000 ng/mL in 3 cases). The mean serum levels of soluble CD163 in the HLH group were significantly higher than in the control and IM groups (P<0.05). The serum levels of soluble CD163 in EBV-positive children were positively correlated with EBV-DNA copies and serum levels of ferritin and LDH, but were negatively correlated with white blood cell count, neutrophil count, hemoglobin and platelet count. The follow-up after treatment for three HLH patients showed that serum levels of soluble CD163 were significantly reduced, but the soluble CD163 levels rebounded in one patient who was complicated by fungal pneumonia infection.</p><p><b>CONCLUSIONS</b>The levels of serum soluble CD163 may be related to the severity in children with HLH. The EBV-positive children with soluble CD163 levels >10 000 ng/mL should be considered the possibility of HLH.</p>

Research Progress on Chemoresistance Mechanism of Nuclear Factor Kappa B Signalling Patheway in Acute Myeloid Leukemia / 中国实验血液学杂志

Acute myeloid leukemia (AML) is the most common leukemia in adult, among them the childhood acute myeloid leukemia accounts for 15% to 20%. After exploring and investigating this disease for 60 years, the systematic chemotherapy can achieve complete remission for 75%-80% AML patients, but only 20%-30% AML patients out of them can be cured, and other AML patients relapsed or died of this disease. The primary and/or seondary chemotherapy resistance may be the main reasons of the poor prognosis in AML. The activity of nuclear factor kappa B (NF-κB) involved into multi-layers of physiological functions. Among them, the activity of NF-κB and the apoptosis toleration associated with the sensitivity to chemotherapy. All these indicated that the inhibition of NF-κB may be a promising direction to reverse chemoresistance and improve chemotherapeutic effects for AML. Herein is the review of recent research progress on the field of the roles of NF-κB activation in AML and its application in AML therapy.

Intracytoplasmic injection of morphologically selected spermatozoa from patients with male factor infertility: clinical and embryo development outcomes / 南方医科大学学报

<p><b>OBJECTIVE</b>To test whether intracytoplasmic injection of morphologically selected spermatozoa (IMSI) from patients with male factor infertility can improve the clinical and embryo development outcomes of intracytoplasmic sperm injection-embryo transfer (ICSI-ET).</p><p><b>METHODS</b>We performed IMSI for 82 couples diagnosed with obstructive azoospermia at high magnification (×6600) and traditional ICSI for another 91 couples using testicular sperms. We also performed IMSI for 44 couples with teratozoospermia at high magnification (×6600) and traditional ICSI for 71 patients using ejaculated sperms. The clinical and embryo development outcomes were compared between the cycles.</p><p><b>RESULTS</b>For obstructive azoospermia, IMSI and ICSI showed no significant difference in the rates of cleavage (95.5% vs 96.7%), D3 top quality embryos (28.2% vs 29.2%), implantation (26.4% vs 32.3%), pregnancy (47.3% vs 50%), blastocyst formation (54.3% vs 54.6%), or abortion (14% vs 7.3%) (P>0.05), but a significantly higher normal fertilization rate was achieved in IMSI group (84.3% vs 77%, P<0.05). For teratozoospermia, the 2 techniques resulted in no significant differences in the rates of cleavage (96.2% vs 95.2%), D3 top quality embryo (27.6% vs 27.1%), implantation (28.2% vs 30.7%), pregnancy (43.7% vs 43.2%), or abortion (9.7% vs 10.5%) (P>0.05), but the normal fertilization rate (68% vs 75.5%) and the blastocyst formation rate (54.6% vs 67.9% ) were significantly higher in IMSI group (P<0.05).</p><p><b>CONCLUSION</b>IMSI can improve the normal fertilization rates in couples with male factor infertility (including obstructive azoospermia and teratozoospermia) and increase blastocyst formation rate in cases of azoospermia.</p>

Application of noninvasive fetal trisomy testing based on massively parallel sequencing for the detection of chromosomal deletions and duplications / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the value of noninvasive fetal trisomy testing based on massively parallel sequencing for the detection of chromosomal deletions and duplications.</p><p><b>METHODS</b>Peripheral venous blood was taken from pregnant women with a high risk. Free fetal DNA in maternal plasma was used for library construction and subjected to massively parallel sequencing. Positive results were validated by traditional karyotype analysis or array-CGH. Phenotype of the fetus was observed through patholoical evaluation.</p><p><b>RESULTS</b>Thirteen out of 629 cases were suspected to harbor chromosomal aberrations, which included 9 aneuploid cases and 4 structural abnormalities. The latter included one case with dup (18q) (14.35 Mb), del (18q) (21.34 Mb), one with dup (3q) (35 Mb) and two with dup (7q) (7.0 Mb). Among these, dup (18q ) (14.35 Mb), del (18q) (21.34 Mb) and dup (3q) (35 Mb) were confirmed by karyotype analysis and patholoical evaluation. However, the two cases with dup (7q) were validated by karyotype analysis and array-CGH as false positives. The phenotype with the fetus also presented as normal.</p><p><b>CONCLUSION</b>The introduction of maternal plasma sequencing for prenatal testing could dramatically improve the efficiency for detecting large, partial (> 10 Mb) chromosomal deletions and duplications.</p>

Analysis of clinical phenotypes of compound heterozygotes of Hb J-Bangkok and β-thalassemia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze hematological characteristics of compound heterozygotes of Hb J-Bangkok and β-thalassemia, and to explore the influence of Hb J-Bangkok on the phenotype of β-thalassemia.</p><p><b>METHODS</b>Peripheral blood samples from a patient carrying Hb J-Bangkok and a β-thalassemia mutation, her family members and three sporadic Hb J-Bangkok carriers were collected. RBC analysis and hemoglobin electrophoresis were performed. Genotypes of α- and β-globin genes were analyzed.</p><p><b>RESULTS</b>The father of the proband and the three sporadic cases were single carriers of Hb J-Bangkok. All of them were asymptomatic and have normal hematological parameters except for an abnormal hemoglobin band detected on hemoglobin electrophoresis. The proband was a compound heterozygote for Hb J-Bangkok and β-thalassemia mutation IVS-Ⅱ-654. She presented typical β-thalassemia trait, featuring hypochromic microcytic anemia and increased Hb A₂ level. An abnormal hemoglobin band was also detected.</p><p><b>CONCLUSION</b>Carriers of Hb J-Bangkok alone are asymptomatic. Co-existence of Hb J-Bangkok and β-thalassemia may not aggravate the phenotype. Therefore, couples with one carrying Hb J-Bangkok and another carrying a β-thalassemia mutation do not require prenatal diagnosis.</p>

Large-scale population-based genetic screening and prenatal diagnosis for thalassemias in Zhuhai City of Guangdong Province / 中华妇产科杂志

ObjectiveTo report the results of preventive control program of severe thalassemias in Zhuhai City of Guangdong Province from 1998 to 2010.MethodsAs the guide centre of marriage and childbearing and the greatest maternity hospital in Zhuhai City of Guangdong Province,Zhuhai Municipal Maternity and Child Healthcare Hospital constructed the genetic screening network for thalassemias testing and referred for follow-up and for genetic counseling.The couples for premarital medical examination or regular healthcare examination in pregnancy were enrolled to this preventive control program.A conventional strategy of screening for heterozygote was used to identify the α- and β-thalassemia traits in women and their spouses according to the standard procedures of hematological phenotype analysis which was recommended by Thalassemia International Federation (T IF).Then those suspected couples at risk were diagnosed for α- and β-thalassemia by PCR-based DNA assays.The couples at risk for severe thalassemias were counseled and offered prenatal diagnosis and termination of pregnancy in case of an affected fetus in the rights of consent and of option voluntarily.ResultsFrom January 1998 to December 2010,85 522 brides and grooms-to-be for premarital screening and 41 503 pregnant women in addition to 14 141 partners for prenatal screening were recorded,the covering rates of premarital screening and prenatal screening in the city were 92.698％ (from 1998 to 2003) and 27.667％ (from 2004 to 2010),respectively.Totally 10 726 cases were found to be the carriers of thalassemias,with 7393 for o-thalassemia (5.237％,7 393/141 166) and 3333 for β-thalassemia (2.361％,3 333/141 166).A total of 257 couples at-risk for severe thalassemias were detected including 190 for α-thalassemia and 67 for β-thalassemia.Among them,251 (97.7％,251/257) couples were performed prenatal diagnosis.During the preventive control program,a total of 72 fetuses with severe thalassemias including hemoglobin H disease were voluntarily terminated.In Zhuhai City,the average annual birth rate of fetuses with severe thalassemia was declined by 32.9％ (49/149).ConclusionsThis study has reduced effectively birth rate of perinatal infants with severe thalassemias in Zhuhai City by genetic screening and prenatal diagnosis of thalassemia in the large population of 13 years.Our summary comes out of technical proposals for prenatal screening and diagnosis,which could be take example by preventative control of thalassemia in other regions of China where are prevalent.

Analysis of hematological characteristics on the 79 co-inheritance of α-thalassemia and β-thalassemia carriers in Guangxi / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the hematological characteristics of co-inheritance of α-thalassemia (α-thal) and β-thalassemia (β-thal) and to survey the incidence of co-inheritance of α-thal and β-thal in Guangxi.</p><p><b>METHODS</b>DNA samples from 370 primary and middle school students who were β-thal carriers in Guangxi were further processed for the α-goblin gene mutation screening, and were grouped based on the genotype of β- and α-goblin gene. The hematological indexes to the different groups were compared by One-way ANOVA.</p><p><b>RESULTS</b>Of the total 370 β-thal carriers, 79 were found to carry α-thal, which gave a frequency of 21.35% for β-thal carriers and 1.36% for coincidence of these two common disorders in the local population. As expected, the 79 patients presented very variable α-globin alterations in combination with β-globin mutations, showing 31 genotype combined with the coincidence of both Hb disorders. Except the genotypes of 3 β-thal heterozygotes combined with ααα(anti3.7) triplication and 2 β-thal carriers with IVS-II-654(C→T)/N combined-α(3.7)/αα presented the phenotype of thalassemia intermedia, and other 74 carriers with co-inheritance of α-thal and β-thal all presented the phenotype of β-thal trait. There were significant differences between β-thal heterozygotes and the carriers with a co-inheritance of both β+α(0) thal in MCH, MCV and Hb. In addition, there existed significant difference between the carriers with a co-inheritance of both β+α(+) thal and a co-inheritance of both β+α(0) thal in MCV, MCH and Hb.</p><p><b>CONCLUSION</b>Compared to that of β-thal heterozygotes, the carriers with a co-inheritance of α-thal and β-thal had slighter phenotype with hematological characteristics. It's difficult to distinguish the double heterozygotes with the co-inheritance of α-thal and β-thal from β-thal heterozygotes by hematological indexes, the molecular diagnosis should be performed.</p>

Effects of acute lymphoblastic leukemia children bone marrow mesenchymal stem cells on drug resistance of K562/A02 cell line / 中国实验血液学杂志

The aim of study was to investigate the effect of acute lymphoblastic leukemia (ALL) children bone marrow mesenchymal stem cells (MSC) on resistance of K562/A02 cells and its mechanism. MSC obtained from bone marrow of AL children were cultured and identified. The co-culture of MSC and K562/A02 and the culture of K562/A02 cell suspension alone was performed, of which 2 kinds of cells were treated with same concentration of adriamycin (ADM), and the rate of apoptosis was detected by flow cytometry, bcl-2 and bax of K562/A02 were detected by RT-PCR, while mdr1 gene level was detected by FQ-PCR. The results indicated that the MSC separation and proliferation were viable and steady. The apoptosis rate of the K562/A02 cells co-cultured with MSC was 1.97 ± 0.11%, while apoptosis rate of the K562/A02 cells cultured alone was 8.38 ± 0.29%, there was significant difference (p < 0.05). As compared with the K562/A02 cells cultured alone, the bcl-2 gene expression in K562/A02 cells co-cultured with MSC obviously increased; ratio of bcl-2/bax was obviously enhanced. The mdr1 gene level in K562/A02 co-cultured with MSC was no statistical different from K562/A02 cultured alone (p > 0.05), which suggested that adhesion co-cultured with MSC did not induce mdr1 expression higher than the culture of suspension. It is concluded that the MSC of ALL children can escape the leukemia cells from proapoptotic effect of drugs, the resistance of K562/A02 to ADM may be involved in enhancement of bcl-2 gene expression of K562/A02 cells co-cultured with MSC, but not in relation to mdr1 gene in K562/A02 cells themselves.

Detecting phospho-signaling protein of bone marrow leukemia cells by phospho-signaling flow cytometry / 中国实验血液学杂志

The purpose of this study was to establish the phospho-specific flow cytometry (phospho-flow) to detect the phosphorylated signaling proteins of leukemia cells and to evaluate its useful value in leukemia study. The bone marrow of leukemia children was collected, and treated by phospho-flow of extracted mononuclear cells (MNC) and phospho-flow of directly fixed bone marrow (BM) respectively. In phospho-flow of extracted MNC, the MNC extracted from BM were fixed and permeabilized, then were cultured with P-AKT and P-ERK1/2, finally were analyzed by flow cytometry. In phospho-flow of directly fixed BM, the BM was treated with fixation/lysis buffer and 90% methanol, then were incubated with the surface CD antibody, P-AKT and P-ERK1/2, finally the treated BM cells were analyzed by flow cytometry. The results showed that the positive rates of P-AKT and P-ERK1/2 in MNC treated by phospho-flow of extracted MNC of 26 leukemia children were 46.2% and 30.8% respectively, while the positive rates of P-AKT and P-ERK1/2 in BM treated by phospho-flow of directly fixed BM were 50.0% and 38.5% respectively. The comparison of positive rates of P-AKT and P-ERK1/2 between the 2 treatment protocol showed no difference (p > 0.05). It is concluded that the phospho-flow of directly fixed BM established by our laboratory can be used to analyze the signaling proteins of leukemia cells.

The changes and significance of T lymphocytes and subsets before and after receiving highly active anti-retroviral therapy in children infected with HIV or AIDS patients / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the changes of the T lymphocytes and their subsets before and after receiving highly active anti-retroviral therapy (HAART) in children who were infected with HIV or AIDS patients.</p><p><b>METHODS</b>Ninety-nine children met the criteria were recruited. All of them had received HAART in Guangxi Center for Disease Control and Prevention from May 2006 to April 2009. Peripheral blood of 2 milliliter was collected before treatment (D0) and after 3, 6, 12, 18, 24, 30, and 36 months (M3, M6, M12, M18, M24, M30 and M36), respectively. Four-color fluorescence flow cytometry was used for the detection of the absolute numbers of CD3(+), CD4(+), CD8(+) T lymphocytes in peripheral blood. And then, the percentages of CD3(+), CD4(+), CD8(+) T lymphocytes in the CD45(+) cells and the ratio of CD4/CD8 were calculated.</p><p><b>RESULTS</b>Sixteen-five (65.66%) cases were treated with lamivudine (3TC)/zidovudine (AZT)/nevirapine (NVP), and 16 (16.16%), 8 (8.08%) and 10 (10.10%) cases were treated with 3TC/stavudine (D4T)/NVP, 3TC/AZT/efavirenz (EFV) and 3TC/AZT/lpv-rtv (LPV/r), respectively. The median of the ratio of CD4/CD8 were 0.39, 0.51, 0.61, 0.65, 0.70, 0.73 and 0.76 in M3, M6, M12, M18, M24, M30 and M36, respectively which were significantly higher than that in D0 (0.19) (Z values were -5.158, -7.375, -9.078, -8.853, -8.373, -5.845 and -5.844 respectively, P < 0.000). The median of CD4% were 16.92%, 22.70%, 25.54%, 26.66%, 27.99%, 30.36% and 29.30% respectively in M3, M6, M12, M18, M24, M30 and M36 respectively, which were also higher significantly than that in D0 (9.92%) (Z values were -5.268, -7.568, -9.496, -9.171, -8.760, -6.190 and -5.964 respectively, P < 0.000). In addition, the median of the absolute numbers of CD4(+)T lymphocytes in peripheral blood were 631, 813, 1050, 946, 1057, 1166 and 894 cells/mm(3) respectively in M3, M6, M12, M18, M24, M30 and M36, which were higher significantly than that of D0 (382 cells/mm(3)) (Z values were -3.318, -5.288, -6.661, -5.886, -5.801, -4.110 and -3.600 respectively, P < 0.000). However, the median of CD8% were 47.25%, 43.01%, 43.04%, 42.60%, 41.37%, 40.83% and 38.31% respectively in M3, M6, M12, M18, M24, M30 and M36, which were lower significantly than that of D0 (53.17%) (Z values were -3.082, -4.697, -5.282, -5.846, -5.757, -3.883 and -4.380 respectively, P < 0.001).</p><p><b>CONCLUSION</b>There is certain rules for the changes of T lymphocytes and their subsets, which may play important roles in the evaluation of the therapeutic effect and the clinical application guidance of HAART.</p>