BackgroundHypoxia-inducible factor-1 α (HIF-1α) specific double-stranded RNA ( dsRNA ) mediated by liposome inhibit reinal neovascularization in mice at dose-dependent manner. ObjectiveThe present study was to investigate the inhibitory effect of dsRNA targeting HIF-1α on retinal neovascularization in mice.MethodsModels of oxygen-induced retinal neovascularization were set up in C57BL/6J mouse through exposure of postnatal day 7 ( P7 ) to (75±3) ％ oxygen for 5 days.Fluorescein conjugated Dextran angiography of retinal vascular was performed to identify the retinal neovascularization.The 8 mice of the normal group were raised in the room air.Fifty-one P7 mice exposed to(75±3)％ oxygen for 5 days and then returned to the room air and assigned to control group ( 3 mice),empty vector group( 3 mice) and gene therapy group (45 mice),and the latter were medially divided to 9 groups randomly according to dose-ratio ( liposomes ∶ plasmid).The pSilencer 2.1-U6 hygro was injected in the model mice of empty vector group,and different dose-ratios of pSilencer2.1-U6 hygro-HIF-1α dsRNA were injected respectively in the model mice of various gene therapy groups.Fluorescein conjugated Dextran angiography of retinal vascular was performed to observe the morphology of new blood vessels,and retinal slides were prepared to score the numbers of nuclei extending beyond the inner limiting membrane( ILM ),and expression of vascular endothelial growth factor(VEGF) was detected in the retina by immunohistochemistry.Results The retinal blood vessels of the normal group formed a fined radial branching pattern.The retinal vascular patterns in the control group and the empty vector group were characterized by decreased central perfusion in both the superficial and the deep layers.The abundant vessels were distorted and irregular in the control group and empty vector group,and the obstructed capillary and lots of neovascular tufts were seen.The retinal neovascularization and non-perfusion distraction in the every gene therapy group were reduced markedly with the most severe appearance in 1 ∶ 1 ( liposomes ∶ plasmid) dose-ratio group.Few vascular endothelial cell nucleus extending beyond the ILM were found in the normal group;while a large number of vascular endothelial cell nucleus were showed in the control group and empty vector group with the occurring rate 100％.Statistically,no significant difference was seen in the number of nuclei extending beyond the ILM between the control group and the empty vector group(11.57±5.85 vs 11.53±6.15),however,that in 1∶1 (liposomes∶plasmid) group was reduced markedly ( 2.17 ± 4.23 ) ( P ＜ 0.01 ).Immunohistochemistry revealed that VEGF was faintly expressed in the normal group but strongly expressed in the control group and the gene therapy group.VEGF expressions of various gene therapy groups were weaker than ones of the control group and the empty vector group.ConclusionsRetinal neovascularization can be efficiently inhibited by intravitreal injection of the pSilencer2.1-U6 hygro-HIF-1α dsRNA mediated by liposome.Proportion of 1 ∶ 1 (liposomes ∶ plasmid)has a maximized efficiency.

Objective To analyze the clinical characteristics of macrophage activation syndrome (MAS). Methods Clinical data was analyzed in 9 children who had been diagnosed as MAS in our hospital from Jan, 2003 to Aug, 2006. Results Seven children were boys, 2 children were girls, aged 5 months to 12 years. Clinical manifestations included long-term fever, hemophagocytic cell in bone marrow, anemia, arthritis, enlargement of lymph nodes, enlargement of liver and spleen, liver dysfunction, abnormal fat metabolism. Two cases had acute respiratory, distress syndrome(ARDS), 2 cases were complicated with multi- pie organ failure(MOF), two patients died. Glucocorticoid combined with immunosuppressive therapy were effective, HP(Plasma Exchange)was applied in one severe case and was shown to he effective. Conclusion MAS is a serious complication of JIA, especially in systemic-onset juvenile idiopathic arthritis. It is very im- portant to recognize and treat MAS earlier.

The cancer stem cells (CSCs) from human osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs were isolated and identified. The primary cells derived from human osteosarcoma were digested by trypsin to prepare a single-cell suspension, and mixed homogeneously into 1.2% alginate gel. Single-cell alginate gel was cultured with serum-free DMEM/F12 medium. Epirubicin (0.8 mug/mL) was added to the medium to enrich CSCs. After cultured conventionally for 7 to 10 days, most of cells suspended in alginate gel were killed by epirubicin. But few cells survived and some single-cell cloning spheres formed. Immunofluorescent staining for Oct3/4 and Nanog was implemented to find cells with properties of self-renewal and multi-potential differentiation. Cells from cloning spheres were transplanted into BALB/c mice to detect the tumorigenicity in vivo. The results showed that some cells positive for Oct3/4 (TRITC) and Nanog (TRITC) were found in single-cell cloning spheres, and most of positive cells were concentrated in the core of sphere. Cells from spheres could form osteosarcoma in the body of mice. It was concluded that cells from single-cell cloning spheres had the properties of the expression of parts of stem cell genes (Oct3/4 and Nanog), resisting anti-cancer drugs, and tumorigenicity in vivo. To sum up, it is believed that cells obtained from osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs are cancer stem cells.

Rapid detection of pathogenic microorganisms is important to the prevention and control of diseases.Com-pared with traditional approaches, electrochemical DNA biosensors present great advantages in promising rapid, portable, sensitive and cost-saving detection of pathogens.In this review, the working principle of electrochemical DNA biosensors and the progress in detection of pathogens is introduced, the latest developments of DNA tetrahedron structure and new nano materials in electrochemical DNA biosensors are reviewed, and the challenges to and prospects of development in this field are also discussed.

Objective To investigate the meaning of expression of apoptosis related molecules NFKBp65 and p53 and GPX1-mRNA in patients with Keshan disease(KSD).Methods Sixteen chronic Keshan Disease patients were enrolled in KSD group according to electrocardiogram,chest X ray film and clinical examinations on 15,September in 2009,and 23 healthy people were included in control group from physical examination taken in The Second Affiliated Hospital of Xi'an Jiaotong University.Fresh blood(5 ml)was collected from antecubital vein of all subjects in the fasting state.Total mRNA and protein of blood sample were isolated using Trizol.GPX Assay Kit was used to detect GPX enzyme activity,and GPX1-mRNA expression was determined by SYBR Real-Time PCR.Meanwhile,expression of apoptosis related molecules NFKBp65 and p53 were determined by Western blot.Results GPX enzyme activity decreased significantly in KSD group[(108.61±14.10)U]compared with control group[(122.78±11.89)U,t=2.874,P<0.05],GPX1-mRNA level of KSD group(0.553±0.299)notably KSD group(0.802±0.057)compared with control group[(1.065±0.355),t=6.829,P<0.01].p53 increased in KSD group(1.604±0.191)compared with control group[(1.137±0.186),t=3.033,P<0.05].Conclusiom Decreased GPX1-mRNA expression may result in lower GPX enzyme activity of patients with KSD.Thus oxidative damage increases and cadioeyte apoptosis is activated by activating apoptosis signal pathway.

OBJECTIVETo establish a new method and validate its feasibilities for quality evaluation of Fructus Epimedii.

METHODFour main effective flavones, epimedin A, epimedin B, epimedin C and icariin were selected as analytes to evaluate the quality of Fructus Epimedii. The relative correction factors (RCF) of icariin to the other three flavones were calculated. The method was evaluated by comparison of the quantitative results between external standard method and QAMS method.

RESULTNo significant differences were found in the quantitative results of three flavones by external standard method and QAMS method.

CONCLUSIONIt is feasible and accurate to evaluate the quality of Fructus Epimedii.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Flavonoids ; analysis ; Reproducibility of Results ; Statistics as Topic ; methods ; Time Factors

Abstract: Objective　To understand the situation of drug-resistant tuberculosis screening and epidemiological characteristics of multi-drug resistant tuberculosis (MDR-TB) and pre-extensively drug resistant tuberculosis (pre-XDR-TB) in Changsha, in order to provide a scientific basis for improving the quality of drug-resistant tuberculosis prevention and control in the city. Methods　Demographic information and drug susceptibility date of etiologically positive pulmonary tuberculosis patients in Changsha from 2018 to 2021 were collected, the successful rate of resistance screening, incidence and tendency in MDR-TB and pre-XDR-TB in patients included in this study were statistically analyzed accordingly. 　　Results From 2018 to 2021, the successful screening rates were 86.2%, 87.7%, 81.9% and 71.5% for MDR-TB and 82.2%, 84.8%, 76.9% and 68.2% for pre-XDR-TB, respectively. In each year, MDR-TB patients identified accounted for 7.6% (101/1 222), 6.5%(124/1 774), 6.6%(110/1 555) and 6.3%(99/1 478), and pre-XDR-TB patients identified accounted for 3.6%(46/1 219), 3.8%(69/1 766), 4.4%(69/1 495) and 4.6%(69/1 436), correspondingly. The incidence of MDR-TB showed a slowly downward trend, while the incidence of pre-MDR-TB showed a slowly upward trend, with neither decreasing nor increasing trends being statistically significant (（χ2=1.947，0.806，P>0.050). The incidence of MDR-TB in the retreatment failure population was 66.6% (2/3), and the others, failure initial treatment and recrudescence populations were 23.5% (19/81), 16.7% (2/12) and 15.2% (70/461), respectively. Similar to the incidence above, the incidence of pre-XDR-TB was 16.7% (2/12) among patients who failed in initial treatment, and 12.2% (9/74), 9.8% (43/439), and 4.5% (2/44) among the others, recrudescence and returned patients, respectively. The incidence rates of MDR-TB and pre-XDR-TB in different populations were significantly different (χ2=117.600，59.030，P<0.05). Conclusions There are still areas for improvement in tuberculosis drug resistance surveillance system in Changsha. On the premise of paying attention to patients in retreatment failure, other, initial treatment failure and relapse patients, high sensitivity molecular drug susceptibility testing, and scientifically efficient screening strategies must be explored.

Humans ; Stress, Mechanical ; Thrombosis ; etiology

0.05.Conclusions The UF effectively removed BUN in sheep with low flow VV-ECMO.The application of UF didn't cause blood shunt in ECMO.The low flow VV-ECMO effectively eliminated carbon dioxide and rerformed oxygenation.

To analyze the genetic characteristics of a polio-I highly variant vaccine recombinant virus in Shandong Province (China) in 2011 and to identify isolates from healthy contacts, two stool specimens from one patient with acute flaccid paralysis (AFP) and 40 stool specimens from his contacts were collected for virus isolation. The complete genome of poliovirus and VP1 coding region of the non-polio enterovirus were sequenced. Homologous comparison and phylogenetic analyses based on VP1 sequences were undertaken among coxsackievirus (CV) B1, CV-B3 isolates, and those in GenBank. One poliovirus (P1/11186), CV-A4 and CV-A8 were isolated from the AFP patient; one CV-A2, Echovirus 3 (E-3), E-12 and E-14, ten CV-B1, and five CV-B3 strains were isolated from his contacts. These results led us to believe that there may be a human enterovirus epidemic in this area, and that surveillance must be enhanced. P1/11186 was a type-1 vaccine-related poliovirus; it combined with type-2 and type-3 polioviruses in 2A and 3A regions, respectively. There were 25 nucleotide mutations with 9 amino-acid alterations in the entire genome. There were 8 nucleotide mutations with 5 amino-acid alterations in the VP1 region compared with the corresponding Sabin strains. Homology analyses suggested that the ten CV-B1 isolates had 97.0%-100% nucleotide and 98.9%-100% amino-acid identities with each other, as well as 92.6%-100% nucleotide and 99.2%-100% amino-acid identities among the five CV-B3 isolates. Phylogenetic analyses on the complete sequences of VP1 among CV-B1 and CV-B3 isolates showed that Shandong strains, together with strains from other provinces in China, had a close relationship and belonged to the same group.
Base Sequence ; Capsid Proteins ; genetics ; immunology ; Child, Preschool ; China ; Humans ; Male ; Molecular Sequence Data ; Phylogeny ; Poliomyelitis ; etiology ; prevention & control ; virology ; Poliovirus ; classification ; genetics ; immunology ; isolation & purification ; Poliovirus Vaccines ; adverse effects ; genetics ; immunology