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.

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.

0.05).Subsequent stratification revealed that MICA*A5 was present at significantly lower frequency in patient group with chronic granulocytic leukemia(CML)(RR=0.635,P=0.038 0);MICA*A4 at significantly lower frequency in patient group with lyphololastic leukemia(ALL)(RR=0.120,P=0.029 7);MICA*A5 at significantly higher frequency in patient group with acute non-lyphololastic leukemia(ANLL)(RR=2.229,P=0.021 8),all compared with control group.Conclusion:MICA-STR allelic variation is associated with leukemia in Hunan Han population;The data also suggest the heterogeneity of MICA-STR allele associated with different clinical types of leukemia.

Objective To investigate the effect of oxcarbazepine(OXC) and sodium valproate(VPA) on electroencephalogram(EEG) and peripheral blood levels of homocysteine(Hcy) and asymmetric dimethylarginine(ADMA) in adult patients with partial epilepsy.Methods From May.2014 to May.2015,a total of 100 patients with partial epilepsy were enrolled and randomly divided into treatment group(treated with OXC) and control group(treated with VPA),with 50 cases in each group.After treatment,changes of EEG indices,Hcy,ADMA,cognitive function and adverse reaction were analyzed.Results Before treatment,there was no significant difference of EEG indices between the two groups(P>0.05).After treatment,the incidence rates of α wave decreasing more than 0.5 Hz,increasing of θ wave and increasing of δ wave were significantly different(P< 0.05).Before treatment,there was no significant difference of serum Hcy and ADMA levels between the two groups(P>0.05).After treatment,serum Hcy and ADMA levels were both significantly increased(P<0.05).Before treatment,Mini-mental State Examination(MMSE) scores of the two groups were without significant difference(P>0.05).After treatment,MMSE score of treatment group was higher than that of control group(P<0.05).In treatment group,there were 1 case of skin rash and 2 cases of gastrointestinal discomfort,which were self-improved.In control group,there were 3 cases of dizziness,5 cases of skin rash and 1 case of gastrointestinal discomfort,which were self-improved.Conclusion The effects of OXC and VAP on peripheral blood levels of Hcy and ADMA could be similar,and compared with VAP,OXC could significantly improve cognitive function in patients with epilepsy.

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.

Colonoscopy has been accepted as the standard method for evaluation of colon and rectum,its success rate depends on the quality of bowel preparation. Aims:To evaluate the cleanliness and tolerance of fractionated dose versus single dose polyethylene glycol electrolyte solution( PEG-ES) bowel preparation regimens for colonoscopy. Methods:A total of 427 consecutive asymptomatic individuals undergoing colorectal cancer screening were enrolled and randomly assigned into 2 groups. Subjects in group A drank 1. 5 L PEG-ES on the eve and 4 hours before colonoscopy, respectively;subjects in group B received a single dose of 3 L PEG-ES 5 hours before colonoscopy. Score and degree of Boston bowel preparation scale(BBPS)and PEG-ES related adverse effects of the two groups were assessed and compared. Results:There were no significant differences in gender,age and cecal insertion rate between group A and group B(P ﹥ 0. 05). Score of BBPS was significantly higher in group A than in group B(P ﹤0. 01). Both regimens met the requirement of conventional colonoscopy,however,the cleanliness of colon was graded as excellent in more subjects of group A( P ﹤ 0. 01),and less subjects of group A complained PEG-ES related nausea(P ﹤0. 05). Logistic regression analysis revealed that the PEG-ES drinking pattern was associated with cleanliness of colon and occurrence of nausea( P ﹤ 0. 05). Conclusions:Fractionated dose PEG-ES regimen provides a better colonic cleansing quality and tolerance for bowel preparation of colonoscopy,which is superior to that of single dose regimen.

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 .

Objective To investigate the signal pathway of platelet activation in acute respiratory distress syndrome (ARDS). Methods Thirty healthy Sprague-Dawley (SD) rats were randomly divided into control group (n = 6) and model group (n = 24). The model of ARDS was reproduced by intravenous injection of oleic acid (0.25 mL/kg), and the rats in control group were injected with the same amount of normal saline. The blood of abdominal aorta was collected at 2, 6, 24, and 72 hours after model reproduction, the platelets were separated, and c-Jun N-terminal kinase phosphorylation (pJNK) levels which was one of major protein kinases in the mitogen-activated protein kinases (MAPKs) signal pathway was determined by Western Blot. The rats were sacrificed, the lung tissues were harvested, and lung coefficient (lung weight/body weight ×100%) and lung wet/dry (W/D) ratio were calculated. Pathological changes in the lung tissue were observed with hematoxylin-eosin (HE) staining in light microscope. Results Comp ared with the control group, platelet pJNK level in ARDS model group was significantly increased at 2 hours after model reproduction (gray value: 0.72±0.09 vs. 0.22±0.01), and peaked at 6 hours (gray value: 0.91±0.03 vs. 0.22±0.01), then it was decreased gradually. It was also significantly higher than that of control group till 72 hours after model reproduction (gray value: 0.39±0.06 vs. 0.22±0.01, all P < 0.05). Lung coefficient and lung W/D ratio in ARDS model group were significantly increased at 2 hours after model reproduction as compared with those of control group [(1.30±0.20)% vs. (0.60±0.10)%, 6.00±0.60 vs. 3.30±0.30], then they were decreased gradually. They were also significantly higher than those of control group till 72 hours after model reproduction [(0.90±0.10)% vs. (0.60±0.10)%, 4.80±0.70 vs. 3.30±0.30, all P < 0.05]. It was showed by light microscopy that lung tissue of rats in the control group had no significant pathological changes. At 2 hours after model reproduction in model group, clearly visible alveolar edema and interstitial edema, interstitial lung infiltration of inflammatory cells, small blood vessels dilation and congestion were found, and the re were a lot of protein exudates. The lesions of lung peaked at 24 hours. At 72 hours, absorption of most of fluid leaking in alveolar, alveolar space narrow, alveolar septum thickening, the reduction of inflammatory cells infiltration, fibrous tissue proliferation, and micro thrombosis formation were found. Conclusion In ARDS, in addition to pathological changes in the lung tissue, platelet activation occurs, and its activation process is related to the priming of JNK signal transduction pathways.