Feature Deconstruction Strategy Based Identification of Acute Features Associated With Impairment of Activities of Daily Living in Patients With Traumatic Brain Injury / 中山大学学报(医学科学版)

ObjectiveTo identify acute phase features associated with the prognosis of traumatic brain injury （TBI）. MethodsThrough two traditional strategies， correlation analysis and prediction model， and one innovative research strategy based on feature deconstruction， a retrospective analysis was conducted using demographic， acute phase and chronic phase features of 354 TBI patients to identify acute phase features associated with activities of daily living （ADL） in chronic phase of TBI. For feature deconstruction strategy， the LASSO （Least Absolute Shrinkage and Selection Operator） algorithm was used to build a prediction model that could effectively predict ADL based on non-ADL chronic phase features. The model could indicate the key chronic phase dimensions determining the ADL in TBI patients. We then identified demographic and acute phase variables that were significantly associated with these key chronic phase features. ResultsThe feature deconstruction strategy revealed that ADL could be deconstructed into chronic phase dimensions such as weak limbs in TBI population. Importantly， to the best of our knowledge， this strategy revealed for the first time the association of these important acute phase features with specific chronic phase impairment features. For example， TBI patients had a higher risk for chronic phase recent memory impairment if they had a prolonged coma time and low GCS scores at acute phase ［scaled coma time OR95%CI = 94.288 （35.095， 273.231）； scaled GCS OR95%CI = 0.068 （0.030， 0.147）］； the patients had a higher risk for insight impairment and disorientation at chronic phase if they had hydrocephalus at acute phase ［insight impairment OR95%CI = 6.760 （3.653，12.855） ； disorientation OR95%CI = 6.538 （3.530， 12.490）］. All strategies showed that the strongest risk factors for ADL damage in the chronic phase included prolonged coma time and low GCS scores as well as hydrocephalus. ConclusionThis study provides an innovative research strategy to establish the association between acute injury features and chronic recovery features， and to identify demographic and acute phase features associated with the prognosis of TBI.

Influencing Factors of Intestinal Metaplasia or Atypical Hyperplasia in Chronic Atrophic Gastritis Patients and Prediction Model Establishment / 中国实验方剂学杂志

ObjectiveTo investigate the influencing factors of intestinal metaplasia or atypical hyperplasia in chronic atrophic gastritis （CAG） patients and establish a prediction model. MethodThe clinical records and laboratory examination data of 335 CAG patients treated in the department of gastroenterology of the Second Affiliated Hospital of the Anhui University of Chinese Medicine from June 2016 to June 2021 were collected. Single and multiple Logistic regression analyses were used to explore the influencing factors of intestinal metaplasia or atypical hyperplasia in CAG patients by SPSS 26.0. A prediction model was constructed based on the data of the related influencing factors. In addition, 115 CAG patients diagnosed in the First Affiliated Hospital of Anhui Medical University from June 2019 to June 2021 were selected as external validation samples to verify and evaluate the prediction efficiency of the constructed prediction model. ResultMultiple Logistic regression analysis showed that pepsinogen Ⅰ［odds ratio（OR） 0.994，95% confidence interval（CI） （0.990，0.999），P<0.05］，the number of focus［OR 6.765，95% CI（3.831，11.945），P<0.01］， and Helicobacter pylori （Hp） infection［OR 0.546，95% CI（0.335，0.888），P<0.05］ were independent risk factors for intestinal metaplasia or atypical hyperplasia in CAG patients（P<0.05）. The formula of the prediction model is as follows：P=-1.558+0.606×Hp infection-0.006×pepsinogen Ⅰ+1.912×the number of focus. The receiver operating characteristic （ROC） curve showed the specific parameters as below： the area under the ROC curve of 0.76，the Youden index of 0.443，the best cut-off value of 0.52，sensitivity of 0.533，and specificity of 0.910. The prediction model was applied to the data of patients in the validation group for validation，and the predictive efficiency of the model was tested by decision curve analysis （DCA）. The results showed that the model had a good fit and high predictive value. ConclusionPepsinogen Ⅰ，the number of focus， and Hp infection are independent risk factors for intestinal metaplasia or atypical hyperplasia in CAG patients. The prediction model constructed based on these factors has a good fit and high predictive value，which can provide references for the classification of CAG patients and the formulation of individual treatment protocols.

Effects of HMGB1 on Proliferation and Secretion of Human Bone Marrow Mesenchymal Stem Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of high mobility group protein 1(HMGB1) on the proliferation and cytokine expression of human bone marrow mesenchymal stem cells (MSC).@*METHODS@#Different concentrations of recombinant human HMGB1 protein (100, 200, 400, 800 and 1000 ng/ml) were incubated with MSC for 24, 48, 72 h and the proliferation of MSC were detected respectively by using the CCK-8 method and flow cytometry. The best concentrations of HMGB1 incubated with MSC was determined (200 ng/ml, 1000 ng/ml), and the flow cytomerty was used to determine the effect of HMGB1 on the proliferation of MSC. The mRNA expression levels of IL-10, TGF- β1, TSG-6 and IFN-γ in MSC incubated with HMGB1 protein were detected by real-time quantitative PCR and ELISA.@*RESULTS@#The result of MSC identification and flow cytometry showed that the CD105, CD73 and CD90 were expressed, but did not expression CD45, CD34, CD11b, CD19 and HLA-DR; CCK-8 showed that HMGB1 at the concentrations of 100 ng/ml, 200 ng/ml and 400 ng/ml could promote the proliferation of MSC incubated for 24, 48 and 72 h as compared with the control group (P<0.05), and the most effective concentration was 200 ng/ml; flow cytometry showed that the compared with the control group, HMGB1 200 ng/ml could induce MSC from G1 phase to S phase to promote the proliferation of MSC; QPCR showed that the mRNA expression of MSC cytokines IL-10, TGF-β1, TSG-6 increased while IFN-γ decreased at the concentration of 200 ng/ml HMGB1 as compared with the control group. ELISA experiments showed that the HMGB1 200 ng/ml acting on MSC for 48 h could significantly promoted the secretion of IL-10, TGF-β 1 and TSG-6(P<0．05), while IFN-γ showed no significant difference as compared with control group.@*CONCLUSION@#Recombinant human HMGB1 can promote the proliferation and secretion of MSC in healthy people.

Effect of AR-V7 on endocrine therapy resistance of prostate cancer / 中国病理生理杂志

AIM:To explore the effect of androgen receptor splice variant 7 ( AR-V7 ) on endocrine therapy resistance of prostate cancer cells and the resistance mechanisms .METHODS:Four prostate cancer cell lines were trans-fected with AR-V7 siRNA ( siAR-V7) using Lipofectamine 2000 kit, and the transfected cells were named as PC 3-siAR-V7, DU145-siAR-V7, LNCaP-siAR-V7 and ArCaP-siAR-V7 cells.The prostate cancer cells transfected with negative con-trol ( NC) siRNA served as negative controls .The expression of AR-V7 at mRNA and protein levels was detected by real-time PCR and Western blot , respectively .The cell viability and cell migration rate were measured by MTT assay and Tran-swell method, respectively.The promoter activity of AR and the protein levels of targeted prostate-specific antigen (PSA) and FK506-binding protein 5 (FKBP5) were monitored by luciferase reporter gene assay and Western blot , respectively. The bicalutamide-resistant cell line, LNCaP-DR, was constructed, and the subcellular localization of AR and AR-V7 pro-teins in LNCaP, LNCaP-siAR-V7 and LNCaP-DR cells was observed by the method of immunofluorescence .The protein in-teraction of AR-V7 and heat shock protein 90 ( HSP90) was determined by co-immunoprecipitation .RESULTS:The mR-NA level of AR-V7 in the 4 prostate cancer cell lines was significantly higher than that in normal prostate epithelial cell line RWPE-1 (P<0.05).The AR-V7 level, cell viability and cell migration rate in the cells transfected with siAR-V7 were notablely lowered compared with the NC siRNA-transfected cells (P<0.05).The cell viability was gradually decreased following with the increase in bicalutamide dose , and down-regulation of AR-V7 expression significantly enhanced the sensi-tivity to bicalutamide (P<0.05).Down-regulation of AR-V7 expression significantly inhibited AR promoter activity and reduced the protein levels of PSA and FKBP5 (P<0.05).The results of immunofluorescence observation showed that most AR and AR-V7 were mainly located in the nucleus , a few AR was located in the cytoplasm , and down-regulation of AR-V7 expression inhibited AR nuclear transport .AR was entirely located in the nucleus and the protein levels of AR-V7 was sig-nificantly increased in the bicalutamide-resistant cells .The interaction of endogenous AR-V7 with HSP90 was found in the prostate cancer cells .CONCLUSION: High AR-V7 level is found in the prostate cancer cells , and down-regulation of AR-V7 expression inhibits the cell viability and migration .High AR-V7 level is related to the bicalutamide resistance .The possible mechanism is that AR nuclear transport is mediated by the interaction of AR -V7 with HSP90 to activate AR signal pathway and regulate targeted gene transcriptional activity , thus resulting in drug resistance .

Effect of AR-V7 on endocrine therapy resistance of prostate cancer / 中国病理生理杂志

AIM:To explore the effect of androgen receptor splice variant 7 ( AR-V7 ) on endocrine therapy resistance of prostate cancer cells and the resistance mechanisms .METHODS:Four prostate cancer cell lines were trans-fected with AR-V7 siRNA ( siAR-V7) using Lipofectamine 2000 kit, and the transfected cells were named as PC 3-siAR-V7, DU145-siAR-V7, LNCaP-siAR-V7 and ArCaP-siAR-V7 cells.The prostate cancer cells transfected with negative con-trol ( NC) siRNA served as negative controls .The expression of AR-V7 at mRNA and protein levels was detected by real-time PCR and Western blot , respectively .The cell viability and cell migration rate were measured by MTT assay and Tran-swell method, respectively.The promoter activity of AR and the protein levels of targeted prostate-specific antigen (PSA) and FK506-binding protein 5 (FKBP5) were monitored by luciferase reporter gene assay and Western blot , respectively. The bicalutamide-resistant cell line, LNCaP-DR, was constructed, and the subcellular localization of AR and AR-V7 pro-teins in LNCaP, LNCaP-siAR-V7 and LNCaP-DR cells was observed by the method of immunofluorescence .The protein in-teraction of AR-V7 and heat shock protein 90 ( HSP90) was determined by co-immunoprecipitation .RESULTS:The mR-NA level of AR-V7 in the 4 prostate cancer cell lines was significantly higher than that in normal prostate epithelial cell line RWPE-1 (P<0.05).The AR-V7 level, cell viability and cell migration rate in the cells transfected with siAR-V7 were notablely lowered compared with the NC siRNA-transfected cells (P<0.05).The cell viability was gradually decreased following with the increase in bicalutamide dose , and down-regulation of AR-V7 expression significantly enhanced the sensi-tivity to bicalutamide (P<0.05).Down-regulation of AR-V7 expression significantly inhibited AR promoter activity and reduced the protein levels of PSA and FKBP5 (P<0.05).The results of immunofluorescence observation showed that most AR and AR-V7 were mainly located in the nucleus , a few AR was located in the cytoplasm , and down-regulation of AR-V7 expression inhibited AR nuclear transport .AR was entirely located in the nucleus and the protein levels of AR-V7 was sig-nificantly increased in the bicalutamide-resistant cells .The interaction of endogenous AR-V7 with HSP90 was found in the prostate cancer cells .CONCLUSION: High AR-V7 level is found in the prostate cancer cells , and down-regulation of AR-V7 expression inhibits the cell viability and migration .High AR-V7 level is related to the bicalutamide resistance .The possible mechanism is that AR nuclear transport is mediated by the interaction of AR -V7 with HSP90 to activate AR signal pathway and regulate targeted gene transcriptional activity , thus resulting in drug resistance .

Relationship between polymorphisms of tumor necrosis factor alpha gene and primary myelodysplastic syndromes / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the association of single nucleus polymorphisms(SNP)of tumor necrosis factor alpha (TNF-α) gene (-308 G>A and -238 G>A genotypes) with susceptibility to primary myelodysplastic syndromes (MDS).</p><p><b>METHODS</b>Two SNPs (TNF-α-308 G>A,TNF-α-238 G>A) of TNF-α gene were detected by Taqman probes in 341 MDS patients and 365 unrelated-healthy controls.</p><p><b>RESULTS</b>Compared to healthy controls, the frequency of TNF-α-308 AA+AG genotype and A allele increased (18% vs 10%, P=0.015, 9% vs 5%, P=0.021, respectively) in refractory cytopenia with multilineage dysplasia (RCMD) patients. There was no correlation of TNF-α-308 G>A genotype and allele frequency between MDS and controls. No difference in the genotype and allele frequency of TNF-α-238 G>A were found between controls and MDS or the subtypes of MDS (P>0.05). We did not find any linkage between plasma level of TNF-α and TNF-α-308 G>A or TNF-α-238 G>A genotype. Statistic differences were observed between platelet count[58(1-611)×10⁹/L vs 90(7-352)×10⁹/L]and bone marrow blasts in MDS patients carrying TNF-α-308 G>A GG and AA+AG genotype (P=0.024, 0.019, respectively).</p><p><b>CONCLUSION</b>TNF-α-308 G>A polymorphism was correlated with susceptibility to MDS-RCMD.</p>

Determination of hematopoietic clonality by detection of multiple X-linked gene exonic polymorphic loci using transcription-based clonality assays / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the frequencies of heterozygosity in X-linked G6PD, P55, BTK, and FHL-1 gene exonic polymorphic loci among Chinese females and the value of determination of hematopoietic clonality by detection of these X-chromosome exonic polymorphisms based on X-chromosome inactivation patterns (XCIP)-transcription-based clonality assays (TCA).</p><p><b>METHODS</b>Genomic DNA was extracted from peripheral blood of 446 Chinese healthy females. Allele-specific PCR (ASPCR) or PCR-restriction enzyme digestion method was applied for detecting G6PD, P55, BTK and FHL-1 polymorphisms. Those heterozygotic loci were used as markers to examine the hematopoietic clonality of bone marrow mononuclear cells by TCA from essential thrombocythemia (ET) patients with JAK2V617F mutation and myelodysplastic syndrome (MDS) patients with abnormal karyotype.</p><p><b>RESULTS</b>Among the total 446 genomic DNA samples, the frequencies of heterozygosity in G6PD, P55, BTK and FHL-1 loci were 12.8%, 29.4%, 52.0% and 46.4%, respectively. About 81.4% of females were heterozygous at one or more loci. All 10 ET patients with JAK2V617F mutation and 2 MDS patients with abnormal karyotype, which were heterozygotic in either locus, had monoclonal/oligoclonal hematopoiesis.</p><p><b>CONCLUSION</b>Clonality detection based on X chromosome inactivation patterns-transcription based clonality assays is applicable to about 80% of Chinese females.</p>

A preliminary study of prognostic value of thrombocytopenia in patients with primary myelodysplastic syndromes / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the prognostic value of thrombocytopenia in patients with primary myelodysplastic syndromes (MDS).</p><p><b>METHODS</b>Four hundred and nineteen primary MDS patients were retrospectively analyzed. Kaplan-Meier method, Log-rank test and COX regression model were used to evaluate factors that influence the prognosis.</p><p><b>RESULTS</b>Two hundred and fifty-six cases (61.1%) had thrombocytopenia (PLT < 100×10(9)/L), one hundred and three cases (24.6%) had severe thrombocytopenia (PLT < 30×10(9)/L). Overall survival (OS) tended to shorten along with the decreasing of platelet count. Univariate analysis indicated that PL < 30×10(9)/L, MCV ≤ 95 fl, LDH ≥ 300 U/L, lymphocyte-like micromegakaryocyte, nucleated RBC PAS positive, IPSS cytogenetic intermediate- and poor-risk were all related with poor prognosis. Moreover, the prognosis of patients with RCMD, RAEB-Ior RAEB-IIwas poorer than that of the other subgroups. Among these parameters, PLT < 30×10(9)/L, MCV ≤ 95 fl, IPSS cytogenetic intermediate- and poor-risk group and RCMD, RAEB-I and RAEB-II had independent prognostic significance in multivariate analysis. Modified WPSS prognostic model was proposed by adopting PLT, MCV, chromosomal karyotype and WHO classification. The OS of patients with low risk, intermediate-1 risk, intermediate-2 risk and high risk were 59, 28, 14 and 4 months, respectively, and there was a statistically significant difference between the groups (P < 0.05).</p><p><b>CONCLUSION</b>Severe thrombocytopenia indicated unfavorable prognosis, in combination with MCV, chromosomal karyotype and WHO classification, a modified WPSS prognostic model was proposed and worked well for prognostic indication in patients with MDS.</p>

JAK2 exon 12 mutations in patients with Philadelphia (Ph) chromosome-negative myeloproliferative neoplasms / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate JAK2 exon 12 mutations in patients with Philadelphia (Ph) chromosome-negative myeloproliferative neoplasms (MPN) and the clinical characteristics of patients with JAK2 exon 12 mutants.</p><p><b>METHODS</b>Allele-specific PCR (AS-PCR) was applied to identify JAK2 V617F mutation. Genomic DNA corresponding to exon 12 of JAK2 gene and epigenetic regulator gene (TET2, ASXL1, EZH2) were amplified by polymerase chain reaction (PCR). Identification of mutants was by direct sequencing and classification of mutation types by sequencing followed by plasmid cloning. SNP genotyping of two 46/1 tag SNPs, rs12340895 and rs10974944, was analyzed using commercially available Taqman assays on the 7500HT real-time PCR instrument according to standard protocols.</p><p><b>RESULTS</b>No JAK2 exon 12 mutation was detected in patients with ET, PMF or JAK2 V617F positive PV. Among 13 JAK2 V617F negative PV patients, JAK2 exon 12 mutation was detected as N542-E543del in 2(15.4%) patients who presented with a phenotype of predominant erythrocytosis and erythroid colonic grown from their bone marrow samples in the absence of exogenous EPO, reduced serum erythropoietin (EPO) level, and no mutations in TET2, ASXL1 or EZH2 genes. One of the affected patients was heterozygous for 46/1 but the second was negative for this haplotype.</p><p><b>CONCLUSION</b>There was no need to detect JAK2 exon 12 mutation in ET, PMF or MPN-U patients without JAK2 V67F mutation. Ph negative MPN patients with JAK2 exon 12 mutations had somewhat unique clinical and laboratory features.</p>

An update on epigenetic regulator gene mutations and pathogenesis of myelodysplastic syndromes / 中国实验血液学杂志

The myelodysplastic syndrome (MDS) is a group of heterogeneous clonal disorders. So far, the etiology and pathogenesis of MDS is poorly understood. Recently, more and more epigenetic regulator gene such as TET2, ASXL1, EZH2, DNMT3A and UTX mutations were detected in patients with MDS: TET2 may convert 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (hmC). TET2 is the most frequently mutated gene in MDS known so far and it may act as tumor-suppressor gene. ASXL1 belongs to the enhancer of trithorax and Polycomb (ETP) gene group. MDS phenotypes may be caused not only by loss-of-function of ASXL1 but also by gain-of-function mutations, overexpression of this gene and so on. EZH2 is a kind of histone methyltransferase. EZH2 is frequently over-expressed in a wide variety of cancerous tissue types, which reveals it has oncogenic activity. While, defined mutations resulted in dysfunction of histone methyltransferase activity, suggesting that EZH2 acts as a tumor suppressor for myeloid malignancies. DNMT3A belongs to the DNA methyltransferases (DNMT) gene family. It may be correlated with abnormal methylation status in patients with MDS. UTX coding protein is a histone demethylase, and UTX can affect cell proliferation as well as cell fate decision. Inactivating UTX mutations are found in multiple cancer types recently. These gene mutations may play key roles in the pathogenesis of MDS, which are summarized in this review.