OBJECTIVE: To investigate the expression levels of EZH2 and KMT2D in patients with diffuse large B-cell lymphoma (DLBCL) and their relationship with pathological features. METHODS: 84 patients with DLBCL treated in our hospital from January 2021 to June 2022 were selected as the study subjects, and clinical characteristics such as sex, age and pathological classification of the patients were collected. Immunohistochemistry was used to detecet the expression of KMT2D and EZH2 proteins in tumor tissue cells of the DLBCL patients. The differential expression of KMT2D and EZH2 in subgroups of different sexes, ages, primary sites, clinical stages, Hans subtypes, etc. were compared. The correlation between the expression of KMT2D and EZH2 protein and BCL-6, CD79A was analyzed and validated through the interaction of protein molecular structures. We followed up and recorded the survival status of the patients for 12 months, and analyzed the factors that affect the mortality of DLBCL patients. RESULTS: The positive rate of KMT2D and EZH2 was high (over 95%) in DLBCL patients. There was no significant difference in the expression of EZH2 and KMT2D among subgroups of different sexes, ages and stages (P >0.05). However, patients with different levels of BCL-6 and CD79A expression showed differences in EZH2 and KMT2D expression (P < 0.05). EZH2 and KMT2D were positively correlated with BCL-6 (r =0.391, r =0.332) and CD79A (r =0.309, r =0.258), respectively, and there were interactions in the protein molecular structures. The risk factors for mortality in DLBCL patients include male sex (OR =1.106, 95%CI : 1.082-1.130, P < 0.001), stage II (OR =1.778, 95%CI : 1.567-2.016, P < 0.001), stage IV (OR =2.233, 95%CI : 2.021-2.467, P < 0.001), EZH2 positive (OR =2.762, 95%CI : 1.304-5.850, P =0.008), BCL-6 positive (OR =7.309, 95%CI : 1.340-39.859, P =0.022), age≥74 years (OR =3.080, 95%CI : 1.658-5.723, P < 0.001), and 63-73 years old (OR =2.400, 95%CI : 1.564-3.682, P < 0.001), while KMT2D positive (OR =0.180, 95%CI : 0.054-0.608, P =0.006) and 41-51 years old (OR =0.406, 95%CI : 0.274-0.603, P < 0.001) were factors which could reduce the risk of mortality. CONCLUSION EZH2 and KMT2D are highly expressed in patients with DLBCL, and they are positively correlated with BCL-6 and CD79A, and affect the prognosis of DLBCL patients.
Humans ; Enhancer of Zeste Homolog 2 Protein/metabolism* ; Lymphoma, Large B-Cell, Diffuse/metabolism* ; DNA-Binding Proteins/metabolism* ; Female ; Male ; Middle Aged ; Adult ; Neoplasm Proteins/metabolism* ; Aged ; Immunohistochemistry ; Proto-Oncogene Proteins c-bcl-6/metabolism* ; Prognosis

The conventional oral drug delivery frequently results in the drug elimination before its complete release due to rapid gastric emptying and short gastrointestinal transport time, thus reducing the bioavailability of drug. In order to maintain an effective concentration of drug in the body and maximize its optimal efficacy, the frequency of administrations often needs to be increased. By contrast, gastric retention drug delivery system (GRDDS), as an innovative method of drug delivery, prolongs the retention time of the drug in the stomach and reduces irritation to the gastrointestinal tract. Consequently, it enhances the bioavailability of drug, reduces dosing frequency for patients and improves treatment adherence. In recent years, domestic and foreign studies have been conducted on gastric retention drug delivery systems. Here, we provide a comprehensive overview of the relevant literature published in recent years, examining their current marketing status, various types, as well as in vivo and in vitro evaluation methods.

Objective:To perform molecular diagnosis and pedigree analysis for one case with α-thalassemia who does not conform to the genetic laws,and explore the effects of a newly discovered rare mutation(HBA2:c.*12G＞A)on clinical phenotypes.Methods:Blood samples of the proband and her family members were collected for blood routine analysis,and the hemoglobin components were analyzed by capillary electrophoresis.The common α-and β-globin gene loci in Chinese population were detected by conventional techniques(Gap-PCR,RDB-PCR).The α-globin gene sequences(HBA1,HBA2)were analyzed by Sanger sequencing.Results:By analyzing the test results of proband and her family members,the genotype of the proband was-α3,7/HBA2:c.*12G＞A,her father was HBA2:c.*12G＞A heterozygous mutation carrier.Conclusion:This study identifies a rare α-globin gene mutation(HBA2:c.*12G＞A)that has not been reported before.It is found that heterozygous mutation carriers present with static α-thalassemia.

Objective:To observe the inhibitory effect of dobutamine on proliferation of FLT3-ITD mutated acute myeloid leukemia(AML)cells and explore the feasibility of dobutamine as a monotherapy or in combination with quizartinib for the treatment of this type of AML.Methods:FLT3-ITD mutant cell lines MOLM13 and MV4-11 were cultured in vitro and divided into control group,dobutamine treatment group,quizartinib treatment group,and dobutamine combined with quizartinib treatment group.Cell viability,ROS levels,and apoptosis rate were detected by CCK-8,Flow cytometry,respectively,as well as the expression of YAP1 protein by Western blot.Results:Both dobutamine and quizartinib inhibited the proliferation of FLT3-ITD mutant AML cell lines.Compared with the control group,the dobutamine group exhibited a significant increase in ROS levels(P＜0.01),an increase in apoptosis rates(P＜0.05),and a decrease in YAP1 protein expression(P＜0.05).Compared with the dobutamine group,the combination of quizartinib and dobutamine significantly reduced cell viability(P＜0.05),increased ROS levels(P＜0.01),and decreased YAP1 expression(P＜0.05).Conclusion:Dobutamine as a monotherapy can inhibit the proliferation of FLT3-ITD mutated AML cells,inducing apoptosis.Additionally,the combination of quizartinib enhances the targeted inhibitory effect on FLT3-ITD mutated AML.The mechanism may involve the inhibition of YAP1 protein expression in AML cells of this type,leading to an increase in ROS levels and exerting its anti-tumor effects.

Objective:To explore the effect of heterozygous deletion of histone methyltransferase Kmt2c gene on the hematological system of mice.Methods:CRISPR/Cas9 technology was used to construct mice model of Kmt2c heterozygous deletion(Kmt2c+/-)and the changes of whole blood cell count in mice were continuously monitored by blood routine test.The clonal expansion ability of bone marrow cells was explored by colony formation assay in vitro and the proportion of primitive hematopoietic cells,including long-term hematopoietic stem cell(LT-HSC),short-term hematopoietic stem cell(ST-HSC),and multipotent progenitor cell in mutant mice was analyzed by flow cytometry.Results:Kmt2c+/-mice model was successfully constructed,and the mRNA expression level of Kmt2c was 28％of that of C57BL/6J mice.The colony formation ability of bone marrow cells of Kmt2c+/-mice in vitro increased with the passage times,and the colony number in the fourth generation was significantly higher than that of control group(P＜0.05).The proportions of LT-HSC and ST-HSC in the primitive hematopoietic cell population of Kmt2c+/-mice was 19.6％±3.3％and 28.9％±4.9％,respectively,which showed an increasing trend compared with 16.9％±2.6％and 18.9％±2.5％in control group,but the difference was not statistically significant(P＞0.05).The white blood cell count of Kmt2c+/-mice gradually increased after 12 weeks of monitoring and reached(9.8±1.0)×109/L at the 14th week,which was significantly higher than(7.3±1.4)× 109/L of control group(P＜0.05).Conclusion:The bone marrow cells of Kmt2c+/-mice have potential of clonal expansion.

Objective To observe the echocardiographic manifestations of infective endocarditis(IE)complicated with valve damage in children.Methods Totally 104 children with IE were retrospectively enrolled and divided into non-damage group(n=34),mild damage group(n=39)and dysfunction group(n=31)according to whether complicated with valve damage and damage's degree.The general and echocardiographic data were compared among groups,and the echocardiographic characteristics of IE complicated with valve damage in children were analyzed.Results Significant difference of the proportion of combining with other congenital heart diseases(excluding bicuspid aortic malformations),the incidence of embolization events during hospital stay,also of endocarditis of left cardiac system were found among groups(all P＜0.05).Pairwise comparison showed that in non-damage group,the proportion of combining with other congenital heart diseases was higher,while the incidence of endocarditis of left cardiac system was lower than those in both mild damage group and dysfunction group(all P＜0.05).The incidence of embolization events during hospital stay in non-damage group was lower than that in dysfunction group(P＜0.05).Among 70 cases of IE complicated with valve damage,mitral valve(30/70,42.86％)was the most common involved valve,mostly presented as valve stenosis(63/70,90.00％).No significant difference of valve involvement site,valve structural lesions nor the incidence of valve stenosis was found between mild damage group and dysfunction group(all P＞0.05).Conclusion IE complicated with valve damage in children mostly involved left cardiac system,and the risk of embolization events was higher than that of IE children without valve damage.Echocardiography could be used as an important method for evaluating the site of valve involvement and the degree of damage.

Objective To observe the manifestations of isolated bicuspid aortic valve(i-BAV)in children using echocardiography.Methods Echocardiographic data of 79 children with i-BAV were retrospectively analyzed,and classification of i-BAV was performed.The patients were divided into complication group(n=50)and non-complication group(n=29)according to the existence of valve and/or aortic involvement or not,and echocardiographic parameters were compared between groups.Results After adjusting body surface area(BSA),in complication group,left ventricular end-systolic diameter/BSA was lower,while left ventricular myocardial mass index group was higher than those in non-complication group(both P＜0.05).Type 0 i-BAV was found in 22 cases,with lat subtype as the most common ones(18/22,81.82％),while Type Ⅰ was observed in 57 cases with L-R subtype as the most common ones(39/57,68.42％).The most common subtype in complication group was also Type Ⅰ L-R(31/50,62.00％),with incidence of valve involvement of 90.00％(45/50),mainly including mild aortic stenosis and/or incompetence(37/45,82.22％),and incidence of aorta involvement of 24.00％(12/50),all with type Ⅰ or Ⅱ aortic widening.Conclusion The most common subtype of i-BAV in children was type Ⅰ L-R,with mild valve damage as the main complication and possibility of left ventricular myocardial remodeling.

The syndrome/pattern of defensive qi deficiency is a common basic syndrome of traditional Chinese medicine in clinical practice. However,there is a lack of standardized and operable diagnostic specifications in practical applications. Based on the previous literature,this study proposed the idea of starting from the elements of the syndrome,qualitative diagnostic criteria for the syndrome/pattern of defensive qi deficiency oriented to the entire region of the disease were constructed based on the two dimensions of " deficient defensive qi failing to consolidate the exterior" and " qi deficiency" and constructing a set of quantitative evaluation criteria as the supporting content for the diagnostic items. The core members of the research group attempted to formulate the draft standard,then reached a consensus through the Delphi method expert questionnaire consultation and the Nominal group technique,and finally evaluated the reliability and validity of the standard through clinical verification to provide ideas for the standardization and normalization of research on syndromes.

Objective To develop and validate a risk prediction model based on early platelet-related parameters for bronchopulmonary dysplasia(BPD)in neonates admitted to the neonatal intensive care unit(NICU),and to facilitate early identification and intervention in high-risk populations.Methods Clinical data of 291 preterm infants with a gestational age(GA)≤32 weeks or a birth weight(BW)＜1 500 g,admitted to the NICU,were retrospectively analyzed.Out of these,214 cases were selected as the modeling group.This group was further categorized into the BPD group(n=76)and the non-BPD group(n=138),based on whether they required oxygen therapy at 28 days post-birth.Perinatal data,platelet-related parameters and other indicators between the two groups.Univariate and multivariate Logistic regression analyses were conducted to identify BPD risk factors,followed by the construction of a nomogram.An additional cohort of 105 preterm infants with GA≤32 weeks or BW＜1 500 g,were used to validate the model.This cohort was divided into the BPD group(n=43)and the non-BPD(n=62)group.Receiver operating characteristic(ROC)curve and calibration curve were used to internally verify the efficiency of the prediction model.Results The Logistic regression analysis identified GA,BW,Apgar score at 5 minutes≤7,invasive ventilation,platelet count(PLT)and mean platelet volume(MPV)as significant factors in the model(P＜0.05).The constructed nomogram was formulated using R language,and the areas under the ROC curve(AUC)for the three models were 0.908,0.931 and 0.918,respectively(P＜0.05).The verification group was verified by Bootstrap.The calibration curve showed a good fit.The internal validation AUC values of the three models were 0.877,0.890 and 0.886,respectively.Conclusion GA,BW,invasive ventilation,Apgar score at 5 minutes≤7,MPV and PLT are key risk factors for BPD onset.The risk prediction model based on these indicators can effectively predict BPD,providing clinicians with a valuable tool for early detection and intervention in the development of BPD.

In this study,the immunological characteristics of the PhoP protein were explored with a two-component system of Mycobacterium tuberculosis(Mtb).Bioinformatics was used to predict the B and T cell epitopes of the PhoP protein.A re-combinant expression plasmid was constructed by PCR analysis of the phoP sequence and cloning into the prokaryotic expres-sion vector pET-28a(+).Competent Escherichia coli BL21 cells were transformed with the recombinant plasmid and expres-sion was induced with IPTG.The recombinant PhoP protein was purified by affinity chromatography.Serum levels of PhoP-specific antibodies in Mtb-infected mice and tuberculosis(TB)patients were analyzed with an ELISA.BALB/c mice were im-munized with the PhoP recombinant protein by intramuscular injection.Sera of mice were collected and antibody titers were detected with an ELISA and specificity was assessed by West-ern blot analysis.Mouse splenocytes were isolated and the pro-portions of IFN-y-positive cells and cytokine levels were detec-ted with an ELISpot and ELISA,respectively.Bioinformatics i-dentified 24 B cell and 11 T cell epitopes of the PhoP protein.A prokaryotic recombinant vector of PhoP was successfully con-structed and the recombinant PhoP protein was obtained by purification.Specific antibody levels to PhoP in sera of Mtb in-fected mice and TB patients increased significantly,with preci-sion of 99.9％and 82.5％,and specificity of 100％,respectively.PhoP protein immunization successfully induced production of specific antibodies in mice.Stimulated by antigens in vitro,IL-2 and IFN-γ levels were significantly increased in the splenocytes of immunized mice.Immunization with the PhoP protein induce a humoral immune response and Thl-dominated cellular immu-nity,indicating that the PhoP protein was immunogenic with diagnostic efficacy for TB.These results lay a foundation to clari-fy the role of PhoP in Mtb infection and application for diagnosis and prevention of TB.