AIM: To study the effect of the radix sophorae flavescentis on cellular immunity in rats with Pneumocystis Carinii Pneumonia (PCP) induced by long-term use of immunosuppressant, and explore the action of traditional Chinese medicine for the immunological regulation and infectious prevention after organ transplantation. METHODS: The experiment was conducted at Department of Pathobiology, Jilin Medical College from May 2005 to March 2006. Forty adult healthy female SD rats were selected from Harbin Medical University (Certification: 02473146) and randomly divided into experiment group and control group, with 20 rats in each. The model of PCP was set up by glucocorticoid injection subcutaneously to SD rats (25 mg once, 2 times/week). The mixture of sophorae flavescentis was given to stomach with tube in experiment group (3 mL/kg, 2 times/day), and was consisted of radix sophorae flavescentis, ash bark, amur cork-tree, malt, milkvetch root and danshen root. Six weeks later, all the rats were anesthetized and broncholveolar lavage fluids were collected.①Alveolar washing fluid was concentrated 10 times and the levels of the soluble interleukin-2 receptor (sIL-2R) were examined by double antigen sandwich ELISA.②Blood was sampled from rat eyes and the count of lymphocytes in peripheral blood were detected.③The percentages of CD3+ and CD4+ subgroups were assessed with erythrocyte chaplet kit sensitized by antigen. RESULTS: All 40 rats were involved in the result analysis without drop.①The count of lymphocytes in peripheral blood in experiment group was significantly higher than that in control group (5.1?1.3)%, (0.8?0.3)%, P

OBJECTIVEThe purpose of this study was to develop RT- PCR assay for Rapidly detect and distinguish between Norovirus genogroup I and genogroup II with a pair of primers.

METHODSA pairs of primers specific to capsid prote in ORF2 gene of G I and G II Norovirus were dsigned according to the published complete genome sequence, with which the RNA of Norovirus was extracted and RT-PCR amplification. The sensitivity, specificity of the RT- PCR assay was estimated and apply it to the detection of Norovirus in clinical specimens.

RESULTSThe results showed that the assay possessed high specificity for Norovirus detection and without any evident cross-reaction with other viruses, including rotavirus, enteric adenovirus and hepatitis A virus. The detection limit of RT-PCR assay for Norovirus G I and G II were up to 100 pg/ml and 10 pg/ml respectively.

CONCLUSIONThe RT- PCR assay provide rapid and sensitive detection of Norovirus G I and G II and should prove to be useful for Norovirus diagnosis in the outbreaks of acute gastroenteritis.

Caliciviridae Infections ; diagnosis ; virology ; DNA Primers ; genetics ; Gastroenteritis ; diagnosis ; virology ; Humans ; Norovirus ; classification ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; instrumentation ; methods

AlM:To investigate the prevalence and related high risk factors of retinal vessels disease of native Tibetan among the aged 40 and above in Maqin county, Qinghai province, China.
METHODS:The cluster sampling method was used to investigate the visual acuity and retinal vessel diseases of the native Tibetan among the aged 40 and above in Maqin county.
RESULTS:Totally 2 511 individuals were underwent the survey, among them, 29 cases (37 eyes) were of retinal vessel diseases, the prevalence was 1. 15%, 21 cases (23 eyes) were retinal vein obstruction (0. 84%), 5 cases (10 eyes) were diabetic retinopathy ( 0. 20%), 3 cases ( 4 eyes) were retinal vasculitis (0. 12%). The blindness and low vision of retinal vessels disease were 23 eyes (0. 92%).
CONCLUSlON:All the hypertension, hyperglycemia, erythrocytosis, high altitude and weight are the high risk factors of retinal vessel diseases which are the main eyes fundus disease could grow blind.

Objectives To investigate the effects of lipoxin receptor agonist BML-111 on IFN-βand IE86 mRNA expression of macrophages infected by human cytomegalovirus (HCMV). Methods Macrophages were infected with HCMV (MOI=0.5), and the cultured cells were randomly divided into control group, HCMV group, HCMV+BML-111 group, and HCMV+MP group. The cells were collected at 0,1,2,4,8 and 12 h after infection, and the levels of IFN-βand IE86 mRNA were tested by real-time PCR. Results Compared with HCMV group, the levels of IFN-βmRNA in HCMV+BML-111 group increased significantly (P < 0.05), while the levels of IFN-βmRNA in HCMV+MP group decreased significantly (P < 0.05); Compared with HCMV group, there were no significant differences of the levels of IE86 mRNA in HCMV+BML-111 group (P>0.05), while the levels of IE86 mRNA in HCMV+MP group increased significantly (P < 0.05). Conclusion BML-111 exerts antiviral activity by promoting the expression of IFN-βmRNA at the early stage of HCMV infection.

BACKGROUND:The anti-inflammation and protective effects of lipoxin have been verified in several immunity-related disease models. Preliminary studies of our research group have shown that, lipoxin receptor agonist BML-111 has negative regulation effects on the human cytomegalovirus (HCMV)-induced immunological injury. However, the effect of BML-111 on the HCMV replication remains unclear.
OBJECTIVE:To observe the influence of lipoxin receptor agonist BML-111 on HCMV replication and proliferation in THP-1 macrophages and human embryonic lung fibroblasts.
METHODS:THP-1 macrophages were infected by HCMV AD169 strain, and were divided into three groups:mock infection, HCMV infection, HCMV+BML-111. The final concentration of BML-111 was 100 nmol/L. cells in each group were col ected at 0, 1, 2, 4, 12, 36, 48 hours, the mRNA levels of IE86 and pp65 in the THP-1 macrophages were tested by RT-PCR method. Human embryonic lung fibroblasts were infected with HCMV (MOI=0.1), and were divided into two groups:HCMV infection and HCMV+BML-111. The patho-morphous changes of human embryonic lung fibroblasts were observed under light microscope, and the cellnumber was measured. The infective virus titer changes in human embryonic lung fibroblasts were examined by plaque assay.
RESULTS AND CONCLUSION:After the macrophages were infected by HCMV, compared with the mock infection group, the mRNA levels of IE86 and pp65 in the HCMV group and HCMV+BML-111 group were increased significantly;compared with the HCMV infection group, the mRNA levels of IE86 and pp65 in the HCMV+BML-111 group were increased significantly in the early stage (within 4 hours) after infection, but the pp65 mRNA levels were decreased significantly in the medium and late stages (24-72 hours) after infection. After human embryonic lung fibroblasts were infected by HCMV, the degree of the patho-morphous in the HCMV+BML-111 group reached 100%2 days earlier than the of HCMV infection group. The infective virus titer reached the peak 2 days earlier than the HCMV infection group, but no significant difference was found between the two groups. BML-111 accelerates the replication of HCMV in the early stage of infection, but inhibits the expression of pp65 gene in the late stage. BML-111 has no impact on the proliferation of the infective HCMV titer in vitro.

Objective To evaluate the therapeutic effects and complications of modified facial mask for non-invasive ventilation (NIV) in elderly patients with respiratory failure.Methods A total of 132 elderly patients(107 males and 25 female,aged 78.5±8.6 years) treated with NIV from February 2008 to May 2011 were randomized into two groups:modified facial mask(group A,n=68,56 males and 12 females,aged 78.8±22.2 years) and control facial mask(group B,n=64,64 males and 13 females,aged 76.6±20.4 years).Duration of NIV,time in RICU(respiratory intensive care unit),length of hospital stay,risk for hospital acquired pneumonia (HAP),risk for invasive ventilation,cure rates,in-hospital mortality,NIV failure rate and cost were compared between the two groups.The complications of NIV,such as oropharyngeal dryness,skin damage of face and nose,abdominal bloating,gas leakage from mask were also compared between the two groups.Results Compared with group B,duration of NIV(12.2±2.3 d vs.18.4±3.6d),time in RICU(7.3±3.2d vs.14.6t5.4d),length of in hospital stay(16.6±4.2d vs.28.2±6.2)d,and cost(2.23±0.12 ten thousand yuan vs.4.23± 0.24 ten thousand yua) in group A were significantly decreased(t=9.72,14.91,13.08,10.81 respectively,all P＜0.05).The risk for invasive ventilation [2.9％ (2 cases) vs.43.8％(28 cases)],NIV failure rate [5.9％ (4 cases) vs.12.5％ (28 cases)] were also decreased in group A compared with group B(x2 =31.26,25.74,both P＜0.05).Compared with group B,The complications of NIV such as skin damage of face and nose[4.4％ (3 cases) vs.37.5％ (24 cases)],abdominal bloating [2.9％ (2 cases) vs.28.1％ (18 cases)],gas leakage from mask [8.8 ％ (6 cases)vs.50％(32 cases)] in group A were significantly decreased(x2 =31.26,25.74,all P＜0.05).Conclusions Modified facial mask for NIV is effective in the treatment of elderly patients with respiratory failure.The complications and in-hospital mortality are reduced with the application of modified facial mask for NIV and it is highly tolerated by patients.Modified facial mask for NIV is the first choice in the treatment in elderly patients with respiratory failure.

To investigate the induction of apoptosis of mouse colonic adenocarcinoma CT26 cells by recombinant Clostridium difficile toxin B (rTcdB), CT26 cells were exposed to different concentrations of rTcd B. Inhibition of cell proliferation was measured by MTT assay. The activation of Caspase 3 was measured by colorimetric method. Cell morphological analysis and flow cytometry were performed to confirm cell apoptosis. rTcd B inhibited the proliferation of CT26 cells in a timeand dose-dependent manner. Caspase 3 activity in CT26 cells was elevated remarkably after rTcd B exposure for 6 h, 12 h, 18 h or 24 h, as compared with the control group. Morphological changes were observed by fluorescence microscopy. The exposure of rTcd B to CT26 cells induced a timeand dose-dependent apoptotic cell death as determined by flow cytometry analysis. The results showed that recombinant Clostridium difficile toxin B induced apoptosis of CT26 cells.

AIM: To investigate whether a pathological micro-environmental culture system consisting of cholestatic sera induces embryonic stem cells (ESC) to differentiate into hepatocyte-like cells in vitro, and select hepatic stem cells from differentiating embryonic stem cells. METHODS: Mouse ESC, E14 cell line, were cultured in Dulbecco's modified Eagle's medium containing 106 U/L recombinant mouse leukemia inhibitory factor (rmLIF) and 10% FCS. After embryonic bodies formed by the hanging drop culture method, they were exposed to fibroblast growth factor-4 (FGF-4) and hepatocyte growth factor (HGF) for one week, and then placed to a pathological micro-environmental culture system consisting of 5% cholestatic sera and cultured for 2 weeks. Morphological examination, immunocytochemical staining of albumin and CK8/18 were carried out, and mRNA level of albumin and transthyretin were detected by RT-PCR. Glycogen storage and urea synthesis of the cells were tested with PAS staining and colorimetric assay, respectively. RESULTS: The proliferation of cells was inhibited at the early stage when cultured in a pathological micro-environmental culture system consisting of 5% cholestatic sera, but 2 weeks later, a large number of epithelial-like cell colonies were observed, which exhibited hepatocellular phenotype, expressing albumin and CK8/18, transcribing mRNA of albumin and transthyretin and synthesizing glycogen and urea. CONCLUSION: A pathological micro- environmental culture system consisting of 5% cholestatic sera could not only induce embryonic stem cells to differentiate into hepatocyte-like cells, but select hepatic stem cells from differentiating embryonic stem cells initially induced by FGF-4 and HGF in vitro as well.

Objective To find the different plasma-associated proteins of rheumatoid arthritis (RA) by using two-dimensional gel electrophoresis for understanding the pathogenesis of RA. Methods The total protein from either RA patients or normal ones was prepared by means of immobilized pH gradient based on two-dimensional gel electrophoresis. After silver staining, gel-image analysis was performed by using PDQuest. The differentially expressed proteins were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS). Results 2-DE patterns of plasma from controls and RA patients were presented. The results showed that average number of protein spots was 592 and 563 respectively, and the corresponding average matching rate was 89% and 87% respectively. Gel-image analysis revealed that there were 24 differential protein spots. A total of 15 differential protein spots were successfully identified by MALDI-TOF-MS, of which 6 proteins were up-regulated as compared with control. Conclusion The differentially expressed proteins can be observed in plasma from RA and controls, which can be used to elucidate the pathogenesis of RA for further study.

Background Nd: YAG laser posterior capsulotomy is an important way for after cataract.Usually the patient will use glucocorticoid eye drops to treat the anterior chamber inflammation after operation,but there is potential risk of elevating intraocular pressure (IOP).Objective This study was to compare the clinical effectiveness and safety of loteprednol etabonate ophthalmic suspension,tobramycin+ dexamethasone eye drops and fluorometholone eye drops following Nd: YAG laser posterior capsulotomy.Methods A randomized-controlled clinical trail was performed.One hundrcd and seventy-onc cycs of 127 paticnts who received Nd: YAG laser posterior capsulotomy for after cataract were randomly divided into four groups.Loteprednol etabonate ophthalmic suspension,fluorometholone eye drops,tobramycin+dexamethasone eye drops and systane eye drops was topically administered respectively in the four groups after laser posterior capsulotomy and 6 times per day for 5 days.IOP was measured with Goldmann tomometer 1 hour before operation and 1 hour,1 day,3 days and 7 days after operation.The ocular anterior segment inflammatory response was examined under the slit lamp and scored based on the Peizeng criteria.Written informed consent was obtained from each patient before any relevant medical procedure.Results The IOP was (18.2 ±4.7),(20.1 ±5.7),(18.7±5.5),(19.0 ±4.1),(19.5 ±3.5) mmHg in various time points in the loteprednol etabonate group; (18.7 ±5.3),(20.9±5.7),(21.3±4.5),(21.0±4.9),(22.5±6.5) mmHg in the fluorometholone eye drops group ; (17.9± 6.3),(20.3 ± 6.1),(23.0 ± 3.7),(24.7 ± 4.9),(24.5 ± 6.5) mmHg in the tobramycin +dexamethasone group and(18.4±6.3),(20.7±3.7),(22.7±6.5),(19.6±4.8),(18.5±3.5) mmHg in the systane group,showing a significant difference among the 4 groups (Fgroup =3.876,P =0.023).With the time lapse,the IOP was gradually reduced in the loteprednol etabonate group and systane group,but that in the fluorometholone group and tobramycin+dexamethasone group was elevated,showing a significant difference among them (Ftime =3.801,P =0.031).No any ocular and systemic adverse effect was found in various groups.The percentage of grade 1 and 2 of aqueous inflammatory cells was lower in the loteprednol etabonate group and tobramycin+dexamethasone group than the fluorometholone group and fluorometholone group and systane group(H =8.276,P =0.012).The percentage of Ⅰgrade of aqueous flare was 8％ in the loteprednol etabonate group,22％ in the fluorometholone group,18％ in the tobramycin+dexamethasone group and 30％ in the systane group,with a significant difference among them (H=9.305,P=0.000).Conclusions The use of corticosteroid eye drops can relieve the inflammatory response of ocular anterior chamber after Nd: YAG laser posterior capsulotomy.Loteprednol etabonate ophthalmic suspension has a better anti-inflammatory effect and less influence on IOP.