Objective: To investigate the role of apoptosis-inducing factor (AIF) in TAp63γ-induced apoptosis. Methods: Plasmid pcDNA3. 1-TAp63γ was transfected into human esophageal squamous cancer EC9706 cells for 24 h. Apoptosis of EC9706 cells was detected by observing DNA fragmentation. The apoptotic rate was measured by flow cytometry. The translocation of AIF in EC9706 cells was studied with Mitochondrial/Cytosol/Nuclear extraction method. The plasmid that expressed hairpin RNA specific for AIF were constructed using chemosynthesis and gene recombination technology, and the interfering efficacy of AIF shRNA was detected by western blotting. After siRNA interference the apoptotic rate of EC9706 cells transfected with pcDNA3. 1-TAp63γ was analyzed by flow cytometry. Results: Agarose gel electrophoresis revealed that DNA ladder, a typical feature of apoptosis, was observed in the pcDNA3. 1-TAp63γ plasmid-transfected cells 24 later, but not in the pcDNA3. 1 plasmid-transfected cells. The apoptotic rate was 13.64% and 1. 37% after EC9707 cells were transfected with pcDNA3. 1-TAp63γ and pcDNA3. 1 plasmid, respectively. Signals of AIF-immunostaining were detected in the nucleus and cytosol proteins of pcDNA3. 1-TAp63γ transfection group, but not in pcDNA3. 1 transfection group. The plasmid expressing shRNA specific for AIF was successfully constructed and the interfering efficiency was about 80%. The apoptotic rate in AIF siRNA group was 4.52% at 24 h after transfection with pcDNA3. 1-TAp63γ. Conclusions: TAp63 γ gene expression induces the apoptosis of EC9706 cells. Down-regulation of AIF protein expression decreases the apoptosis induced by TAp63 γ.

Abstract: Objective To explore the clinical data of children with Talaromyces marneffei (TM) infection admitted to the pediatric intensive care unit (PICU) to improve the diagnosis and treatment of TM infection in children. Methods The clinical data of 23 children with TM infection treated in four PICUs in southern China from January 2013 to December 2022 were retrospectively analyzed, including clinical characteristics, laboratory tests, treatment regimens, and outcomes. Results Among the 23 children, there were 14 boys and 9 girls, with a median age of 24 months. The time from onset to admission was 15 (range 10-30) days, and one patient had immunodeficiency disease. Common symptoms included fever (91.3%, 21 cases), cough (78.3%, 18 cases), and hepatosplenomegaly (78.3%, 18 cases). Severe clinical complications included multiorgan dysfunction (69.6%, 16 cases), septic shock (65.2%, 15 cases), and acute respiratory distress syndrome (65.2%, 15 cases). All patients elevated CRP levels, and 69.2% (9/13) had a positive G test. Elevated IgE levels were observed in 53.3% (8/15) cases, CD4/CD8 inversion in 17.6% (3/17) cases, and reduced NK cells in 84.2% (16/19) cases. HIV tests were negative in all cases. TM was most frequently detected by blood and bone marrow cultures. Seven cases were diagnosed with immunodeficiency by genetic testing. With monotherapy or combination of amphotericin B or/and voriconazole, followed by oral medication, 13 (56.5%) children died. Conclusions TM infection is clinically atypical in HIV-negative children, and patients admitted to PICU have rapidly deteriorated, with severe complications and a high mortality rate. Early use of multiple samples, and multiple methods to detect TM, combined with immune function and genetic test, is helpful to early diagnosis. The antifungal treatment strategies still need further study.

Objective To evaluate the antitumor effect of corosolic acid and its impact on CAM and YSM angiogenesis.Methods The effects of CRA on A549 proliferation was studied by MTT method in vitro.Tumor-bruden nude mice model were established by injecting A549 lung cancer cell marked with bioluminescent to nude mice,and the growth of tumor in mice were detected by IVIS small animal in vivo imaging system.Experiment of chicken embryos eggs are used to observe the role of drugs on CAMand YSMblood vessels. Results The value of IC50 of CRA for A549 in vitro was 26.8μg/mL.A tumor-burdened animal experiment results showed that the CRA to A549 solid tumor has a certain therapeutic effect.CAM and YSM blood vessels of chicken embryos eggs treated with CRA were decreased significantly than negative control group.Conclusion CRA has certain therapeutic effect for A549,which may be related to the inhibition of angiogenesis in tumor tissues.

Water soluble extract (WSE) is an important index for the quality evaluation of Astragali Radix (AR). In this study, the WSE of the wild AR from Shanxi province (SX) and the cultivated AR from Gansu Province (GS) were compared. The WSEs of two types of AR were determined according to the appendix of Chinese pharmacopoeia. Then the WSEs were subjected to NMR analysis, and the obtained data were analyzed using HCA, PCA, OPLS-DA, microarray analysis, and Spearman rank analysis. In addition, the Pearson correlation of differential metabolites were also calculated. The results showed that the WSE content of GS-AR (37.80%) was higher than that of SX-AR (32.13%). The main constituent of WSE was sucrose, and other 18 compounds, including amino acids, organic acids, were also detected. Multivariate analysis revealed that SX-AR contained more choline, succinic acid, citric acid, glutamate, taurine and aspartate, while GS samples contained more sucrose, arginine and fumaric acid. In addition, the Pearson correlations between different metabolites of the two types of AR also showed apparent differences. The results suggested that the WSE of two types of AR differs not only in the content, but also in the chemical compositions. Thus, the cultivation way is important to the quality ofAR. This study supplied a new method for the comparison of extract of herbal drugs.

Background and purpose: Uveal melanoma (UM) is the most common primary intraocular malignancy in adult. Due to a high tendency for early metastasis the treatment of UM is very difficult. This study aimed to explore an effective approach for the treatment of patients with UM, we designed a strategy that combined HtrA2 gene therapy and radiation therapy. Methods:pIRES-Egr1-Omi/HtrA2 (pEgr1-HtrA2) recombinant plasmids were constructed and transfected into human UM cells (OCM-1) in vitro. The transfected cells were exposed to irradiation. HtrA2 mRNA and protein levels were detected by qRT-PCR and Western blot respectively. Assays that evaluated the apoptosis inducibility caused by HtrA2 gene therapy combined with radiation was performed by lfow cytometry. Followingly, the effects of HtrA2 overexpression on the in vitro radiosensitivity of uveal melanoma cells were investigated by clonogenic formation assay. The in vivo effects of HtrA2 gene therapy combined with radiation therapy were evaluated in different groups. Results:The recombinant plasmids could be successfully transferred into OCM-1 cells and transfection of pEgr1-HtrA2 plasmids combined with radiotherapy caused dramatically elevation of HtrA2 compared with non-irradiation cells in mRNA and protein levels, which was associated with increased apoptosis.Furthermore, we observed that the transfection of pEgr1-HtrA2 could significantly enhance radiosensitivity of OCM-1 cell in vitro. In mice bearing xenograft tumors, pEgr1-HtrA2 combined with radiation therapy signiifcantly inhibited tumor growth compared with the other treatment groups (P<0.05). Conclusion:Our ifndings indicate that radiation-inducible gene therapy may have potential to be a more effective and speciifc therapy for uveal melanoma because the therapeutic gene can be spatially or temporally controlled by exogenous radiation.

Objective: To compare two methods (microbial assay and radioimmunoassay) for measuring plasma folate concentrations, and to examine the relationship between plasma folate levels, and alcohol consumption, tobacco use and body mass index, and the risk of hyperhomocysteinemia in China. Methods: We used a microtiter plate microbial assay and a radioimmunoassay to measure the folate concentration in 88 plasma samples. After comparing the results of these two methods and fitting a regression line, we examined the geographical, seasonal, and gender differences in folate concentration of plasma collected from 2 422 adults in south and north areas in China, and evaluated the association of plasma folate concentration, with alcohol consumption, cigarette smoking, and body mass index, and with the risk of hyperhomocysteinemia, using the data from the two assays. Results: The data from the two assays had a linear relationship ( r =0.879, P =0.000); the regression was Y =0.683 X +0.308 (where X and Y were nature logarithmic transformations of plasma folate by microbial assay and radioimmunoassay, respectively); however, the mean plasma folate levels by microbial assay were much higher than those obtained by radioimmunoassay. Both data sets showed similar plasma folate distributions among Chinese adults, associations with other risk factors, and the risk of hyperhomocysteinemia. We estimated that 19.9% of the Southerners and 67.1% of the Northerners had plasma folate concentrations by radioimmunoassay lower than the 6.8 nmol/L used to define plasma folate deficiency. Conclusion: There is a linear relationship between plasma folate levels determined by microbial assay and radioimmunoassay, but because of the different levels obtained in the two assays, it is difficult to use the microbial assay results to evaluate folate status at this time. The use of 10.5 nmol/L as a cut off for plasma folate deficiency by microbial assay needs further study.

Objective To evaluate the clinical application of sweep pattern visual evoked potential in assessing visual acuity (SPVEP-A)of amblyopic children.Design Prospective clinical study.Participants 80 patients(148 case)of amblyopic children and 26 children(52 case)with normal vision.Methods The correlation were analysed of SPVEP-A and E acuity in 80 amblyopic children and of SPVEP-A and PVEP-A in 26 children of normal visual.SPVEP-A was recorded with UTAS-E 3000(LKC)using gratings of 25 differ- ent spatial frequency from 0.2 to 18.8cpd as stimulus.The response were averaged and DFT on the monitor display.SPVEP-A was de- termined by extrapolate to 0 response amplitude.Both transient PVEP-A and SPVEP-A were examined in 26 normal children.The ob- jective visual acuity in PVEP were determined with the highest spatial frequency which evoked a recognizable response.Main Out- come Measures SPVEP-A,PVEP-A and E acuity.Results SPVEP-A were from 0.35 to 0.9 among 80 patients with corrected E acu- ity from 0.1 to 0.8.The correlation coefficient of SVEP-A-I with E acuity was 0.602 .The correlation coefficient of SPVEP-A.between reproducibility detection was 0.448.The correlation between SPVEP-A and E acuity in amblyopic slightly was higher than middle and severe amblyopia(r=0.773 vs r=0.590).In 26 normal children a correlation coefficient of 0.679 was obtained between E acuity anti PVEP-A but 0.424 between E acuity and SPVEP-A.Higher evaluation or lower evaluation of E acuity was found in SPVEP-A in this study.Acuity was upostimated in lower scope of E acuity and underestimated in higher scope of E acuity.The reproducibility and vari- ation were not satisified.Conclusion The sweep VEP is a objective method of amblyopic screening.There is significant positive corre- lation between SPVEP-A and E,but its variability is more great,so we should improve its stability and positive rate uheriorly.

The history and present status of phytoplasma classification are introduced briefly in this paper.The newly classification methods and rules for the description of Candidatus species are reviewed.The key problems and direction on the classification and identification of phytoplasmas in China are discussed.

Objective To discuss the application value of shear wave elastography (SWE) and superb microvascular imaging (SMI) in pathological grading of breast invasive ductal carcinoma (IDC).MethodsSixty patients with breast tumor were selected,which were totally 64 lesions.Before operation,SWE and SMI pattern were started to detect the lesions.And images in SWE and SMI pattern were saved.All lesions were proved to be IDC and pathological gradings were got according to the pathological result.The mean of Young′s modulus(AveT1) and maximum of Young′s modulus(AveT2) were recorded in SWE pattern,and degree of blood was recorded in SMI pattern followed Adler′s grading standard.Then differences of AveT1,AveT2 and degree of blood in different pathological grading of invasive ductal carcinoma were evaluated.Results ①In pathological grading 1 group,the average of AveT1 was (62.1±10.4)kPa,and the average of AveT2 was (93.0±20.1)kPa.In pathological grading 2 group,the average of AveT1 was (79.8±8.6)kPa,and the average of AveT2 was (120.6±18.7)kPa.In pathological grading 3 group,the average of AveT1 was (92.6±12.1)kPa,and the average of AveT2 was (137.3±21.5)kPa.The AveT1 and AveT2 were gradually growing with higher pathological grading of breast invasive ductal carcinoma,which were statistically significant (P<0.05).②In pathological grading 1 group,there were 7 cases in the degree of blood 0-Ⅰ and 14 in the degree of blood Ⅱ-Ⅲ.In pathological grading 2 group,there were 3 cases in the degree of blood 0-Ⅰ and 18 cases in the degree of blood Ⅱ-Ⅲ.In pathological grading 3 group,there were only 1 case in the degree of blood 0-Ⅰ and 21 cases in the degree of blood Ⅱ-Ⅲ.There were more lesions of rich blood with higher pathological grading of breast invasive ductal carcinoma,which was statistically significant (P<0.05).Conclusions There are differences of elastography and degree of blood in different pathological grading of breast invasive ductal carcinoma,SWE and SMI can prompt pathological grading and provide important clinical reference value.

Objective Carbon dots (CDs) are an emerging carbon nano-material which is environmentally-friendly, economical , efficient and stable .Their fluorescence properties can match the semiconductor quantum dot .Moreover , CDs have more excellent biocompatibilities .The purpose of this experiment is to apply CDs to the fluorescent immune probe to make them a new label , which can replace the traditional fluorescent dyes .Methods Using microwave heating method , the high strength fluorescent carbon dots were prepared .Wtih the EDC coupling method , the high strength fluorescent car-bon dots could bond with Escherichia coli antibodies to form a complex immune fluorescent probe .Specific recognition exper-iments were carried out in the model of E.coli O157∶H7.Results CDs were successfully applied to immune recognition of E.coli O157∶H7 and multicolor fluorescence was observed .Conclusion CDs can serve as a label of the fluorescent im-mune probe , and are expected to become a new type of low toxicity biosensor with independent intellectual property rights .