Objective To observe ultrastructural changes in a clinical isolate of Penicillium marneffei(PM) before and after treatment with amphotericin B or voriconazole by using scanning electron microscopy (SEM)and transmission electron microscopy (TEM). Methods A microdilution method was performed to determine the minimum inhibitory concentration (MIC)of amphotericin B and voriconazole against a clinical isolate of PM. Then, the PM isolate was treated with amphotericin B or voriconazole at their MICs and 10-fold MICs for 24, 48 and 72 hours. The ultrastructural changes in this isolate before and after the treatment were observed by using SEM and TEM. Results After the treatment with amphotericin B, SEM showed that the conidia or yeast cells of the PM isolate were gradually damaged, and their outer layers experienced detachment, shrinkage, breakage and adhesion with the increase in treatment duration and concentrations of amphotericin B; TEM also showed degenerated mitochondria, broken nuclei and cell walls, and shrunken cytoplasmic membrane with disappearance of cytoplasmic organelles. Similarly, the damage, shrinkage, shriveling and collapse of PM cells were seen by using SEM, and TEM showed many high-density electron-dense granules in cytoplasm, degeneration of mitochondria, roughening of cell wall surface, damage and shrinkage of cytoplasmic membrane, and disappearance of cytoplasmic organelles after voriconazole treatment. Conclusions Amphotericin B and voriconazole both had a strong antifungal effect on PM, and could induce evident ultrastructural changes, which were positively associated with treatment duration and concentrations. Moreover, amphotericin B caused more severe damage to PM compared with voriconazole.

AIM: To explore whether inhibition of NF-?B by antioxidant pvrrolidine dithiocarbamate (PDTC) sensitizes leukemia cells to cytotoxic drugs and its mechanism. METHODS: The indirect immunofluorescence method and electrophoretic mobility shift assay (EMSA) were used to measure the activation of NF-?B. The apoptotic cells were evaluated by flow cytometry (FCM) and the in vitro growth inhibitory effect was performed using a MTT assay. RESULTS: EMSA showed that NF-?B was activated by daunorubicin (DNR), VP-16 and then was inhibited by PDTC in a dose-dependent manner. NF-?B activation was further verified because of subunit RelA of NF-?B locating in the nuclei. FCM analysis showed that apoptotic index of HL-60 cells was up to (8.97?0.81)%, (16.01?1.06)%, (22.96?1.33)% from (5.34?0.62)%, (10.16?0.42)%, (17.32?1.15)% after exposure of HL-60 cells to 2.5-10 mg/L VP-16 combined with PDTC. VP-16 added with PDTC produced greater growth inhibitory effect to HL-60 cells than did VP-16 or DNR only (P

OBJECTIVE:To study on the phlorotannins in Ecklonia kurome. METHODS:Silica gel column chromatography and Sephadex LH-20 column chromatography were conducted for isolation and identification of phlorotannins,and the compound structures were analyzed and identified based on physicochemical properties and spectral data. RESULTS:Totally 7 phlorotannins were isolated from E. kurome,namely Phloroglucinol(1),Eckol(2),Fucodiphloroethol G(3),Phlorofucofuroeckol A(4),1-(3′, 5′-dihydro-xyphenoxy)-7-(2″,4″,6-trihydro-xyphenoxy)-2,4,9-trihydroxydibenzo-1,4-dioxin(5),Dieckol(6)and 6,6′-bieckol (7). CONCLUSIONS:Compound 5 and 7 are isolated from E. kurome for the first time,and the study has laid a foundation for its quality evaluation.

ObjectiveTo investigate the relationship between HLA-DRB1 alleles and chronic urticaria with positive autologous serum skin test (ASST) in Guangxi Zhuang Autonomous Region.MethodsASST was conducted in 144 patients with chronic urticaria,who were subsequently divided into two groups according to the test result:positive group (n =62) and negative group (n =82).PCR amplification with sequence-specific primers was used to determine the genotypes of HLA-DRB1 alleles in the patients and 199 normal human controls.Chi-square test was performed to analyse the difference in the frequency of HLA-DRB1 alleles between the 3 groups by using the SPSS 13.0 statistical software package.ResultsThere were significant differences in the frequency of HLA-DRB1*01,*1401 and *16 alleles among the patients with positive and negative ASST and the controls (x2 =10.92,Pc =0.032;x2 =35.34,Pc ＜ 0.01 ;x2 =12.69,Pc =0.032).Paired comparison revealed significant differences in the frequency of HLA-DRB1*1401 allele between the patients with positive ASST and controls(RR =17.09,Pc ＜ 0.01 ) as well as between the patients with positive and negative ASST (RR =7.20,Pc ＜ 0.01).ConclusionHLA-DRB1*1401 allele may be,or be linked to,the predisposing gene of chronic urticaria with positive ASST in Guangxi Zhuang Autonomous Region.

Objective To detect the DMD gene mutation sites and the regions of breakpoints in Duchenne/Becker muscular dystrophy (DMD/BMD) patients in northern China. Methods Multiplex amplifiable probe hybridization (MLPA) was used to detect the mutation in 59 cases (51 cases with DMD and 8 with BMD) from northern China and dystrophin gene mutations in their parents. Results From northern China and dystrophin gene mutations 59 families found gene deletions in 33 cases of 59 DMD/BMD patients (55.9%), duplications in 6 cases (10. 2%) and point mutation in one case (1.7%). Intron 44 was most frequently affected (n = 13, 33.3%), followed by intron 50 (n = 11, 28.2%) and intron 45 (n=8, 20.5%). The novel mutations were identified, in two patients including two independent duplications carried by patient D1 149 and a point mutation [5208del(A)] carried by patient D1 65, which were not included in Leiden database. In addition, an exon 22 deletion was found in one patient, which was the first reported case in Chinese patients. Conclusions Deletions are mostly located in the hotspot between exon 45 and 50. Duplications mostly occurred in the 5' end of the gene. Intron 44 is the most frequently affected breakpoint in northern Chinese population.

To explore the effect of NF-κB on bcl-x gene transcription in extended drug resistance leukemia cell line HL-60/E6, drug-resistant subline HL-60/E6 was derived by intermittently exposing HL-60 cells to 6 ng/ml epirubicin. Indirect immunofluorescence was used to demonstrate the location of NF-κB-RelA in HL-60/E6 cells. FCM analysis and RT-PCR were used to detect the efficiency of liposome-mediated ODN transfection and the change of bcl-xL mRNA levels after 5 μmol/L phosphorothioate (PS)-derivatized antisense (AS) oligodeoxynucleotide (ODN) directed to RelA was transferred into HL-60/E6 cells. The results showed that RelA remained persistently active and located at the nuclei of HL-60/E6 cells，but in the cytoplasm of HL-60 cells, the efficiency of liposome-mediated ODN transfection was significantly higher than that of null ODN (P<0.01 in 4 h, 6 h, 12 h, 24 h). Exposure of HL-60/E6 cells to 5 μmol/L AS-PS-ODN directed to RelA led to a maximal 40 % decline of bcl-xL mRNA levels within 8 h. The inhibition rate of bcl-xL mRNA was (15±1.79) %, (28±2.34) %, (40±3.47) %, (20±1.54) % in 4 h, 6 h, 8 h, 15 h， respectively, but it was less than 15 % in control group. It was concluded that NF-κB was involved in regulating bcl-x transcription. It was suggested that NF-κB was an important factor for drug resistance in leukemia cells.

Objective To investigate the relationship of the expression of vascular endothelial growth factor (VEGF)and the mi-crovessel density (MVD)with multi-slice spiral CT perfusion imaging.Methods 80 patients with solitary pulmonary nodules under-went perfusion scan by 16-slice spiral CT.Among them,45 diagnosed as lung cancer by pathology were enrolled in the study.After surgery,the slice of the pecimen was selected similar to the corresponding slice of CT images,and the immunohistochemical staining was performed to determine the expression of VEGF and the MVD.Spearman correlation analysis was used to determine the rela-tionship of expression of VEGF and the MVD with CT perfusion parameters.Results There was more expression of VEGF and the MVD in NSCLC.There were positive correlations between VEGF,MVD and BF (both P<0.05).The peak enhancement image (PEI)and TTP have significant correlations with MVD (P<0.05).Conclusion Some parameters of lung CT perfusion imaging are correlated with MVD and VEGF.

ObjectiveTo test the susceptibility of Penicilliosis marneffei (PM) isolates from Guangxi bamboo rats and patients to voriconazole and several commonly used antifungal agents.MethodsAccording to the Clinical and Laboratory Standards Institute(CLSI) M27-A2 and M38-A document,a microdilution method was used to determine the minimum inhibitory concentration(MIC) of voriconazole,itraconazole,terbinafine,amphotericin B,and fluconazole against mycelial phase (25 ℃) and yeast phase (37 ℃) of 14 PM isolates from Guangxi Bamboo rats and 25 PM isolates from patients.The difference in MIC of the antifungals was assessed by two-sample t test between Bamboo rat PM isolates and clinical PM isolates,and by paired t test between the mycelial and yeast phase of PM isolates.Results The MIC ranges of voriconazole,itraconazole,terbinafine,amphoteriein B and fluconazole were 0.0313-0.1250,0.1250-1.0000,0.0313-0.5000,0.2500-4.0000,2.0000-8.0000 mg/L,respectively for mycelial phase of Bamboo rat PM isolates,0.0078-0.2500,0.0313-0.5000,0.0313-1.0000,0.2500-2.0000,1.0000-8.0000 mg/L,respectively for yeast phase of Bamboo rat PM isolates,0.0313-0.2500,0.0625-1.0000,0.0313-1.0000,0.2500-4.0000,2.0000-32.0000 mg/L,respectively for mycelial phase of clinical PM isolates,0.0039-0.2500,0.0313-0.5000,0.0313-2.0000,0.1250-2.0000,2.0000-16.0000 mg/L,respectively for yeast phase of clinical PM isolates.None of the PM isolates was resistant to any of the antifungals.The MIC of voriconazole was found to be the lowest for PM isolates from both Bamboo rats and patients at the same temperature (37 ℃ or 25 ℃),followed by itraconazole,terbinafine,amphotericin B and fluconazole.Statistical difference was found in the MIC values of itraconazole,terbinafine,amphotericin B between the yeast and mycelial phase of the same PM isolate,but not found in antifungal MIC values between Bamboo rat isolates and clinical isolates at the same phase.ConclusionsOf the tested drugs,voriconazole shows the strongest antifungal potency. The PM isolates from Guangxi Bamboo rats are similar to clinical PM isolates in the sensitivity to voriconazole,itraconazole,terbinafine,amphotericin B and fluconazole.The phase of PM isolates may affect their susceptibility to itraconazole,amphotericin B and terbinafine.

To explore the effect of NF-κB on bcl-x gene transcription in extended drug resistance leukemia cell line HL-60/E6, drug-resistant subline HL-60/E6 was derived by intermittently exposing HL-60 cells to 6 ng/ml epirubicin. Indirect immunofluorescence was used to demonstrate the location of NF-κB-RelA in HL-60/E6 cells. FCM analysis and RT-PCR were used to detect the efficiency of liposome-mediated ODN transfection and the change of bcl-xL mRNA levels after 5 μmol/L phosphorothioate (PS)-derivatized antisense (AS) oligodeoxynucleotide (ODN) directed to RelA was transferred into HL-60/E6 cells. The results showed that RelA remained persistently active and located at the nuclei of HL-60/E6 cells，but in the cytoplasm of HL-60 cells, the efficiency of liposome-mediated ODN transfection was significantly higher than that of null ODN (P<0.01 in 4 h, 6 h, 12 h, 24 h). Exposure of HL-60/E6 cells to 5 μmol/L AS-PS-ODN directed to RelA led to a maximal 40 % decline of bcl-xL mRNA levels within 8 h. The inhibition rate of bcl-xL mRNA was (15±1.79) %, (28±2.34) %, (40±3.47) %, (20±1.54) % in 4 h, 6 h, 8 h, 15 h， respectively, but it was less than 15 % in control group. It was concluded that NF-κB was involved in regulating bcl-x transcription. It was suggested that NF-κB was an important factor for drug resistance in leukemia cells.

OBJECTIVE: To carry out amniocyte karyotyping analysis and chromosomal microarray analysis (CMA) for women with anomalies revealed by fetal echocardiography. METHODS: From January 2019 to December 2021, genetic testing was carried out for 205 fetuses including 97 with soft marker anomalies and 108 with structural heart abnormalities. Among these, 138 only had abnormal fetal echocardiography, whilst 38 and 29 were complicated with extracardiac soft marker anomalies and extracardiac structural malformation, respectively. RESULTS: No significant difference was detected in the detection rate of genetic anomalies between fetuses with heart-related soft markers and those with abnormal heart structures (P > 0.05). Compared with those with abnormal fetal echocardiography alone, the detection rates of chromosomal aneuploidies in those with abnormal extracardiac soft markers or abnormal extracardiac structures were significantly higher (P < 0.05). Twenty-eight chromosomal aneuploidies (including a rare mosaicism), 2 balanced translocations and 1 supernumerary marker chromosome were detected by karyotyping analysis. Twenty-seven aneuploidies, 19 copy number variations (CNVs) and 1 uniparental disomy were detected by CMA. CONCLUSION Prenatal diagnosis has attached great importance to the suggestive role of fetal heart-related soft markers, and chromosomal aneuploidies are more common among fetuses with abnormal extracardiac soft markers and extracardiac structural abnormalities. Chromosomal Karyotyping is useful for the detection of balanced translocations and mosaicisms. CMA is helpful for the detection of CNVs. Identification of the genetic causes can facilitate genetic counseling for the affected couples.
Pregnancy ; Female ; Humans ; DNA Copy Number Variations ; Prenatal Diagnosis ; Fetus ; Echocardiography ; Aneuploidy ; Mosaicism ; Translocation, Genetic