Bronchoalveolar Lavage ; methods ; Humans

Adult ; Bronchoalveolar Lavage ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; therapy ; Retrospective Studies ; Treatment Outcome

Objective To investigate the effects of hepatitis B virus X (HBX) gene on cell morphology and transdifferentiation of human proximal tubular epithelial cells.Methods The eukaryotic vector pcDNA3.1-myc-HBX containing HBX gene was transiently transfected into HK-2 cells by lipofectamine mediation.The expression of HBX was confirmed by Q-PCR and Western blotting.Untransfected HK-2 cells and those transfected with empty vector were used as control.The morphology of HK-2 cells was observed by microscopy,the expressions of differentiation marker proteins α-SMA and E-cadherin were detected by Western blotting and Q-PCR,and the contents of IL-1,IL-6 and TNF-α in the supernatant were detected by ELISA assay.Results HBX was successfully expressed in HK-2 cells after transfection.After transfection of HBX gene,the shape of HK-2 cells became irregular,HK-2 cells significantly expressed E-cadherin and α-SMA,and had high levels of IL-1,IL-6 and TNF-α in the cell supernatant (P＜0.01).Conclusion Overexpression of HBX gene in renal tubular epithelial cells may damage cell morphology and promote the occurrence of epithelial-mesenchymal transdifferentiation,which may be related to the inflammatory microenvironment.

Objective To evaluate the perinatal outcome of three types of monochorionic diamniotic (MCDA) twins with selective intrauterine growth restriction (sIUGR).Methods From January 2005 to June 2012,clinical data of 42 pairs of MCDA twins (84 fetuses) with sIUGR and 71 pairs of normal MCDA twins (142 fetuses) in the same period were analyzed retrospectively in the First Affiliated Hospital of Sun Yat-Sen University.Fetuses with sIUGR were classified into three groups based on umbilical artery Doppler flow.There were 25 cases of type Ⅰ,11 cases of type Ⅱ and 6 cases of type Ⅲ.The perinatal outcome was compared between sIUGR and normal MCDA twins,and among the three types of sIUGR as well.Perinatal outcomes included gestational age at delivery,rate of intrauterine fetal death (IUFD),birth weight,intertwin discordance of birth weight,neonatal death and survival rate at 6 months.Results (1) The gestational age of sIUGR at delivery was significantly earlier than the control group [(34 ± 3),(36 ±2) weeks,respectively],and the rate of IUFD of both fetuses of sIUGR was significantly higher (4.8％,0,respectively).In the sIUGR group,the average birth weight of large or small twins[(2130 ±.350),(1520 ±400) g,respectively] was smaller than those in the control group [(2470 ± 500),(2340 ± 460) g,respectively].The difference was statistically significant (P ＜ 0.05,P ＜ 0.01,respectively).The intertwin discordance of birth weight in sIUGR group was significantly larger (27.6％) than the control group(4.0％,P＜0.01).(2) The gestational age at delivery in type Ⅱ and type Ⅲ [(34 ±5),(34 ±2) weeks,respectively] was significantly earlier than the control group (P ＜ 0.05).The rate of IUFD of both fetuses in type Ⅱ (18％) was significantly higher than in type Ⅰ (0) and the control group (0,P ＜ 0.05).In sIUGR group,the average birth weight of small twins in type Ⅰ,type Ⅱ and type Ⅲ was (1640 ±430),(1330 ±310) and (1500 ±380) g respectively,all of which were significantly smaller than that in the control group (P ＜ 0.05).The average birth weight of small twins in type Ⅱ was smaller than in type Ⅰ and the difference was statistically significant (P ＜ 0.05).In sIUGR group,the intertwin discordance of birth weight in type Ⅰ,type Ⅱ and type Ⅱ was 24.1％,34.6％,31.3％ respectively,all of which were significantly larger than that in the control group(4.0％,P ＜ 0.05).There were no statistically significant differences of the intertwin discordance of birth weight among the three types of sIUGR(P ＞0.05).Survival rate at 6 months in type Ⅱ (64％) was significantly lower than in type Ⅰ (92％) and the control group (91.5％,P＜0.01).Conclusions The perinatal outcome of MCDA twins with sIUGR is poor.The outcome is different among the three types of sIUGR,and type Ⅰ is the worst.Type Ⅱ is associated with a high risk of intrauterine fetal demise.It is important to monitor the intrauterine situation closely.

Objective To investigate the effect of hepatitis B virus X (HBX) gene on apoptosis and immune molecules of human proximal renal tubular epithelial cell line (HK-2).Methods The eukaryotic vector pcDNA3.1-myc-HBX containing HBX gene was transiently transfected into HK-2 cells by lipofectamine mediation.Untransfected HK-2 cells and those transfected with empty vector were used as controls.The TLR4 expression was detected by real-time PCR and Western blotting.The apoptosis of cells and expression of MHC-Ⅱ and CD40 were detected by flow cytometry,and the contents of IL-4 and IFN-γ in the supernatant were detected by EIISA.Results Compared with control groups,the number of apoptotic cells was significantly increased in the HBX transfection group (P ＜ 0.05),and the expressions of TLR4,MHC-Ⅱ and CD40 were also significantly increased in the HBX transfection group (all P＜0.05).IFN-γ level in the supernatant of HBX transfection group was higher (P ＜ 0.05),but IL-4 level was lower as compared to control groups (all P ＜ 0.05).Conclusions Over-expression of HBX gene may induce apoptosis of HK-2 cells and upregulate the expression of immune molecules of renal tubular epithelial cells leading to injury of cells and dysfunction of immunomicroenviroment.

AIM:To construct adenovirus vector expressing mice B7-H1 gene, transfect dendritic cells ( DCs ) , and to study the therapeutic effect of modified DC on thyroid-associated ophthalmopathy ( TAO) in mice.
METHODS: We designed and constructed B7-H1 gene adenovirus expression vector, and transfected DCs from mouse bone marrow, tested the phenotype and function of modified DCs, identificated its negative regulation to immune responses. The modified DCs were infected the sicked mice. And then the immunotherapeutic effect of modified DCs to TAO were tested.
RESULTS: B7 - H1 gene adenovirus vector was constructed and transfected DCs from bone marrow. The titer of the recombinant adenovirus was 1. 8í109 PFU/mL. B7-H1 gene modified DCs characteristics of regulatory DCs, could inhibit positive immune responses. The inhibition proceeding of TAO into mice infected modified DCs, was obviously prior to the control mice. The gene modified DCs, maybe become the new immunotherapy biological agent to thy TAO.
CONCLUSION: We constructed the expression of mouse B7 - H1 gene adenovirus expressed vector successfully, transfected DCs, by vector have properties of regulatory DCs, inhibiting positive immune response and the occurrence and development of thyroid eye disease. Gene modified DCs, reveal potent to the treatment of thyroid eye disease.

Abstract: Schistosomiasis, an important zoonotic parasitic disease, is one of the six major tropical diseases identified by WHO, and also one of the most important parasitic diseases for prevention and control in China. After more than 70 years of efforts, the prevention and control of schistosomiasis in China has made great achievements, and the current epidemic of schistosomiasis in China has entered an extremely low epidemic state, but the distribution base of the only intermediate host of schistosomiasis, Oncomelania hupensis, is still large. For now, the techniques used to monitor schistosomiasis have shortcomings such as time-consuming, laborious and low sensitivity, which cannot meet the current needs of China. Environmental DNA (eDNA) refers to DNA that can be extracted from environmental samples (such as soil, water or air) without isolating any target organisms, which is a complex mixture of genomic DNA and its degradation products from different organisms in the same environment. eDNA technology can reflect the community or species composition information in the ecosystem through DNA extraction and detection of environmental samples. Compared with traditional biological monitoring methods, eDNA technology has the advantages of high efficiency, high sensitivity and environmental friendliness. eDNA has been successfully used for the specific detection of Schistosoma mansoni, Schistosoma haematobium and Schistosoma japonicum. This paper reviews the current detection methods of eDNA, the application and technical limitations of eDNA technology in schistosomiasis monitoring, aiming to provide scientific reference for research in the field of schistosomiasis surveillance.

ObjectiveTo observe the effects of human IL-10 gene transfection on the mRNA and protein expressions of IL-1β and TNF-α in the penumbra area following focal cerebral ischemia-reperfusion injury in rats and to investigate its neuroprotective mechanism. MethodsRats were divided into four groups: normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfected group. The mRNA and protein expressions of IL-1β and TNF-α in the penumbra area were detected by fluorescence real-time quantitative PCR and ELISA respectively. ResultsIn normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfected group, the levels of protein expression of TNF-α in penumbra area were(0.66±0. 04) ,(1.16±0.26),(1. 155±0. 26)ng/g and(0. 84±0. 05)ng/g, and the levels of protein expression of IL-1βin penumbra area were(0.37±0.05), (1.25±0.39), (1.21±0.57) ng/g and(0.62+0.05)ng/g, respectively. Compared with normal control group, the levels of protein expression of TNF-α and 1L-1β were significantly higher in other three groups(all P＜0. 01), and lower in human IL-10 gene transfected group than in ischemic control group and empty plasmid group(all P＜0. 01). In normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfectedgroup, the levels of mRNA expression of TNF-α in penumbra area were 1.00 ±0.53,9.42±1.83,9.69±1.96 and 3.53±1.09, and the levels of mRNA expression of IL-1β in penumbra area were 1.00 ±0.51,27. 81±4.84,23.96 ± 4.90 and 13.55± 4.45, respectively. Compared with normal control group, the levels of mRNA expression of TNF-α and IL-1β were significantly higher in other three groups(all P＜0. 01), and lower in human IL-10 gene transfected group than in ischemic control group and empty plasmid group(all P＜0. 01). ConclusionsThe human IL-10 gene transfection may play an protective effect on cerebral ischemia through inhibiting mRNA and protein expression of IL-1β and TNF-α in the penumbra area following focal cerebral ischemia-reperfusion in rats.

OBJECTIVE To determine the characterization, anti-tumor efficacy and pharmacokinetics of bufalin- loaded PEGylated liposomes compared with bufalin entity. METHODS Bufalin- loaded PEGylated liposomes and bufalin- loaded liposomes were prepared reproducibly with homogeneous particle size by the combination of thin film evaporation method and high pressure homogenization method. The particle size and zeta potential of the liposomes were determined by dynamic light scattering technique. The direct imaging of morphology of liposomes was charactered by transmission electron microscope. The content of bufalin in liposomes was analysed by HPLC method. The entrapment efficiency and the particle size was applied to assess the stability profile, after storage at 4℃ on day 0, 7, 15, 30 and 90. The in-vitro release behaviours of bufalin from liposomes were conducted using dialysis bag technique at 37℃. In-vitro cytotoxicity studies were carried out using MTT〔3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide〕assay on several kinds of tumor cell lines including SW620, PC-3, MDA-MB-231, A549, U251, U87 and HepG2. In-vivo pharmacokinetic study of bufalin liposomes was evaluated by HPLC method. RESULTS Their mean particle sizes were 127.6 nm and 155.0 nm, mean zeta potentials were 2.24 mV and - 18.5 mV, entrapment efficiencies were 76.31% and 78.40% , respectively. In- vitro release profile revealed that the release of bufalin in bufalin- loaded PEGylated liposomes was slower than that of bufalin-loaded liposomes. The cytotoxicity of blank liposomes has been found within acceptable range, whereas bufalin-loaded PEGylated liposomes showed enhanced cytotoxicity to U251 cells compared with bufalin entity. In-vivo pharmacokinetics indicated that bufalin-loaded PEGylated liposomes could extend eliminate half-life time of bufalin in plasma in rats. CONCLUSION The results suggested that bufalin-loaded PEGylated liposomes improved the solubility and increased the drug concentration in plasma.