BackgroundMonocyte chemotactic protein-1 (MCP-1)plays an important role in the tumor,inflammation,diabetic retinopathy and other neovascular disease,but the expression and the role of MCP-1 in the oxygen induced retinopathy(OIR) model have rarely been reported. Objective This study was to investigate the expression of MCP-1 in the retina development of newborn mouse and in mouse models with OIR.Methods C57BL/6J newborn mice were divided into two groups and 60 mice in each group.Mice in OIR group were exposed to 75％ oxygen for 5 days and then to room air.All mice in normal control group exposed to room air only.Ten mice in each group were randomly chosen and sacrificed at postnatal 5,7,12,14,17,21 days.The expression of MCP-1 in mouse retina was detected with the method of immunohistoehemistry and reverse transcription polymerase chain reaction(RT-PCR).Results MCP-1 positive cells were seen in normal mouse retina.Up-regulation of MCP-1 positive cells was detected both in 12 days in normal control group and in 14 days in OIR group.MCP-1 mRNA was detected in mouse retina at 5 days,and a transient up-regulation of MCP-1 mRNA was observed in 12 days in normal control group.MCP-1 mRNA in OIR group significantly increased in 14 days in comparison with the normal control group( P =0.028,P =0.001 ). Conclusions Expression of MCP-1 is detectable in whole retinal development procession of mice.A transient up-regulation of MCP-1 expression is detected in the critical period of retinal vascular development in mice models with OIR,which is closely related to the retinal vascular development and progression of retinal new vessels.

Background Light stimulation at different wavelength influences the development of eyes.It has been showed that blue light can inhibit the growth of eyeball.To study whether blue light exposure can delay the extension of myopia is an interested research project.Objective This study was to investigate the effect of blue light with short wavelength on ocular growth in form deprived myopia (FDM) in guinea pigs and provide a new option for the prevention and treatment of myopia.Methods Thirty-six 2-week-old guinea pigs were reared in the environment of white light.The right eyes of the animals were occluded to establish the FDM models.The models were randomized into the deocclusion + blue light exposure group,simple deocclusion group and continuous occlusion group according to the random number table.The right eyes of the models were deoccluded for 1 hour per day to give the blue light (430 nm) irradiation in the deocclusion + blue light exposure group,and the right eyes were deoccluded for 1 hour per day only in the simple deocclusion group.In the continuous occlusion group,the right eyes of the models were occluded until the end of this experiment.Anterior chamber depth (ACD),lens thickness (LT) and vitreous cavity depth (VCD) were measured by A-type sonography.The binocular diopter of the guinea pigs was detected using retinoscopy in the mydriatic condition.In the fourth week after experiment,the retinal sections were prepared for the regular histopathological examination,and the scleral tissues next to 1 mm from optical nerve were exacted to obtain the dry weight of scleral tissues.Results In the right eyes of the animals,no significant differences were found in the diopter,ACD,LT and VCD before experiment among the 3 groups (all at P＞0.05).At the end of experiment,the refraction of right eye in the deocclusion + blue light exposure group,simple deocclusion group and continuous occlusion group was (+1.11±0.17)D,(+0.90±0.15)D and (-2.73±0.19)D respectively,with a significant difference among them (F=1 445.470,P=0.000).The VCD in the three groups was (3.70±0.09) mm,(3.78±0.11) mm and (3.91 ± 0.08) mm,respectively,showing a significant difference (F =13.243,P＜0.01).In addition,the dry weight of sclera tissues was (0.61 ±0.09)mg in the deocclusion + blue light exposure group,(0.54± 0.08)mg in the simple deocclusion group and (0.43 ± 0.07)mg in the continuous occlusion group,with a significant difference among the 3 groups (F=10.458,P＜0.01).However,there were no significant differences in the ACD and LT among the 3 groups (F=0.203,0.084,both at P＞0.05).Moreover,in the left eyes,no significant differences were found in the diopter,ACD,LT and VCD before experiment among the 3 groups (all at P＞0.05);while at the end of the experiment,the diopter of the continuous occlusion group was significantly lower than that of the deocclusion + blue light exposure group and simple deocclusion group (all at P＜0.05).No significant differences were seen in the ACD,LT,VCD and dry weight of sclera among the 3 groups (all at P＞0.05).Retinal structure was normal in the left eyes of various groups.However,the retinas were thinner in the right eyes of the deocclusion + blue light exposure group with clear layers; while atrophy of the outer segment of photoreceptor and disorder of cell arrangement were seen in the right eyes of the continuous occlusion group.Conclusions During sensitive period of visual development,blue light stimulation can arrest the extension of posterior sclera and elongation of vitreous cavity,which restrains development of myopia.This blue light at the wavelength of 430 nm is safe to retina.

Background Various studies have suggested that inflammatory factors such as leucocytes and macrophages are involved in the occurrence and development of diabetic retinopathy (DR),and many cytokines promote the occurrence of DR.However,the relationship of aqueous and serum monocyte chemotactic protein-1 (MCP-1) and macrophage migration inhibitory factor (MIF) change with DR is unclear.Objective This study was to investigate the effects of MCP-1 and MIF in aqueous and serum during DR development.Methods Eighty patients with type 2 diabetes were enrolled from Beijing Shijitan Hospital.These patients received phacoemulsification or phacoemulsification and vitrectomy from September,2010 to June,2011.Twenty-six cataract patients in the same stage (without diabetes) who underwent phacoemulsification surgery served as controls.According to the clinical stage of the DR,the diabetic patients were classified as the non-DR group (NDR) (20 eyes),non-proliferative DR group (NPDR) (38 eyes) and proliferative DR group (PDR) (22 eyes).Aqueous humour and periphery blood samples were collected during the operation to detect MCP-1 and MIF using enzyme-linked immnunosorbent assay (ELISA).Written informed consent was obtained from each subject before any relevant medical examination.Results The average aqueous MCP-1 levels were(1660.78±562.98),(1463.26± 623.41),(686.76±186.16) and(494.35±148.59) ng/L in the PDR group,NPDR group,NDR group and control group,respectively,showing a significant difference among the 4 groups (F=37.968,P=0.000).No significant differences were found in the aqueous MCP-1 levels between the control group and NDR group (P=0.169),or between the NPDR group and PDR group (P=0.117).However,the aqueous MCP-1 levels were significantly elevated in the PDR group,NPDR group and NDR group compared with the control group (P=0.000).The average aqueous MIF levels were (6.85±1.99),(3.56±0.90),(1.10±0.48) and (0.86 ± 0.46) μg/L,respectively,with significant differences among them (F =144.502,P =0.000).Multiple comparisons between groups were found to be significantly different (P =0.000) according to the LSD-t test,except between the control group and NDR group (P =0.475).A significant positive correlation was seen between the aqueous MCP-1 level and MCP-1 level in all study participants (r =0.564,P =0.000).However,serum levels of MCP-1 and MIF were not statistically significantly different among the 4 groups (F =2.158,P＞0.05;F =0.813,P＞0.05).Conclusions The increase of the aqueous MIF and MCP-1 levels is associated with the progression of diabetic retinopathy.The results suggest that MIF and MCP-1 promote the occurrence of DR.

Objective To investigate SCCmec genotypes and drug-resistance profiles of the methieillin-resistant Staphylococcus epidermidis (MRSE) strains isolated from the patients suffered from diabetic foot infections (DFI) in the Tianjin Metabohc Diseases Hospital. Methods After dabridement, specimens of 390 infectious diabetic foot ulcers in the hospital from Jan 2008 to Jun 2010 were collected from the wound basal parts by cotton swab for culture. The disk-diffusion method was performed to examine antimicrobial susceptibility. DNAs of the MRSE strains were extracted, and their SCCmec genotypes were identified by PCR. Results Twenty of the seventy(28.6% ,20/70)Staphylococcus epidermidis strains were mecA posifive. Among the MRSE isolates, 2 ( 10.0% )were SCCmec Ⅱ ,9 (45.0%)were SCCmecⅢ and 9 (45.0%)were SCCmec Ⅳ. None of the isolates were genotyped as SCCmec Ⅰ or Ⅴ. No mater which genotypes they were, all the MRSE isolates were multi-drug resistant. They were resistant not only to β-lactams (including penicillins, cefoxitin and cephems), but also to non-β-lactams (including macrolides, fiuoroquinolones and sulfonamides ) . Resistance to voncomycin and rifampicin were not found in these strains . Conclusion SCCmec Ⅲ and SCCmecⅣ are major genotypes of the MRSE isolates from the infectious diabetic foot ulcers.The SCCmec Ⅳ genotype strains with multi-drug resistant profiles are prevalent in the diabetic foot infections.

Objective To investigate clinical features and antibiotic resistance of pseudomonas aeruginosa (PA) strains isolated from patients with diabetic foot infections (DFI) in Tianjin Metabolic Diseases Hospital.Methods Eighty-five PA strains were isolated from 428 patients with diabetic foot in the hospital from Jan 2008 to Dec 2010.The clinical features of patients were summarized.Relationships between the isolates and depth of ulcer or severity of infection were analyzed.The disk-diffusion method was performed to examine antimicrobial susceptibility.Results Gram positive (G+) and Gram negative (Gˉ) isolates were 50.47％ and 41.12％,respectively.Multidrug-resistant PA composed 32.9％ of the total PA isolates.The size of ulcers with PA infections was bigger than those with non-PA bacterial infections (P＜0.05).Compared to G+ strains,patients with PA strains were older,had lower hemoglobin,but higher serum sensitive C-reactive protein; and more frequently,they had ischemic ulcer and osteomyelitis.Compared to G+ strains,the PA strains were more frequently isolated from deeper ulcers and with more serious infections(P＜0.05).The resistant rates of PA to cephalosporins,fluoroquinolones,and aminoglycosides were between 32.9％-61.2％,37.6％-42.4％,and 37.6％-62.4％,respectively.Only one out of 85 PA strains was imipenem-resistant.However,sensitiveness of all PA isolates to cefoperazone and sulbactam reached 100％.Conclusion PA strains are mainly found in patients with deeper ulcers and more serious infections.Multidrug-resistant PA is common in DFI.It is important to isolate pathogens and determine their antibiotic resistance correctly in diabetic foot patients in order to provide appropriate drug administration and to reduce the production and dissemination of drug resistant strains.

OBJECTIVE To investigate the neuroprotective effect and possible mechanisms of lute-olin-7-O-β-D-glucuronide (LGU) against focalcerebral ischemic injury. METHODS The focal cerebral ischemic injury model was established by middle cerebral artery occlusion (MCAO). Male Sprague Dawley rats were randomly divided into sham group,model group(MCAO),LGU group(0.24,0.72 and 2.16 mg·kg-1)and positive control group(Edaravone at 5 mg·kg-1).LGU was injected intravenously 30 min after MCAO.Neurological severity score,infarct volume and brain water content were detected 24 h after MCAO and the levels of Na+-K+ATPase,Ca2+ATPase,TNF-α and IL-1β were detected to explore the possible mechanisms.For the therapeutic time window test,LGU(0.72 mg·kg-1)was injected intrave-nously 0.5, 2, 4, 6, 8, 10 and 12 h respectively after MCAO. To evaluate motion behavior, LGU were injected intravenously 30 min after MCAO and once per day during detection period. The changes of motor coordination were detected by rotating rod method and grip strength analysis, and the changes of gaits were detected using DigiGait Imaging System. RESULTS LGU improved the neurological severity score, infarct volume ratio and brain water content. The therapeutic time window of LGU for cerebral infarction and brain edema was at least 6 h and for neurological dysfunction was 12 h.LGU also prolonged the latency on rotarod, increased the forelimb tension and improved 8 gait parameters, including stance duration,stride length,stance width,paw area,paw area variability,gait symmetry,ataxia coefficient and tau propulsion.Furthermore,LGU increased Na+-K+-ATPase and Ca2+-ATPase levels in the cortex and hippocampus in the ischemic side,reduced the levels of TNF-α and IL-1β in the serum. CONCLUSION LGU has a significant neuroprotective effect against cerebral ischemic injury via improving energy metabolism and reducing inflammation.

OBJECTIVE To investigate the protective effect and mechanisms of luteolin-7-O-β-d-glucuronide (LGU) on oxygen glucose deprivation (OGD)-induced H9C2 cardiomyocytes injury. METH-ODS The protective effect of LGU on OGD-induced H9C2 cardiomyocytes death were investigated by MTT assay. The microfilament change of H9C2 cardiomyocytes was detected by phalloidin staining and the lactate dehydrogenase (LDH) leakage rate was also detected by LDH kit. In order to explore the possible mechanisms of LGU, ATP content, intracellular Ca2+fluorescent intensity and concentra-tion, mitochondrial membrane potential (MMP)and the expressions of apoptosis-related proteins were detected by ATP kit,CLSM(Fluo-3/AM probe),Ca2+kit,CLSM(JC-1 probe)and western blotting meth-od, respectively. RESULTS The inhibition of H9C2 cardiomyocyte survival rate inducedby OGD was improvedby pretreated with LGU in a concentrationdependent manner. The microfilaments injury as well as the increase of LDH leakage rate were also improvedby pretreated with LGU.The ATP content was significantly decreased,intracellular Ca2+fluorescent intensity and concentration were significantly increased and the MMP was significantly decreased 4 hafter OGD. LGU significantly reversed the de-crease of intracellular ATP content,the increase of Ca2+fluorescent intensity and concentration and the decrease of MMP.The release of cytochrome C,the expressionsof caspase-9 and caspase-3 in H9C2 cardiomyocytes were increased 16 h after OGD.LGUsignificantly inhibited the changes of these apop-tosis-related proteins. CONCLUSION LGU has a significant protective effect against OGD-induced H9C2 cardiomyocytes injury through inhibiting calcium overload,increasing ATP content,improving mi-tochondrial function and inhibiting apoptosis.

OBJECTIVETo Evaluate four kits for screening HIV antibody by comparing and analyzing the HIV antibody screening positive results and Western Blot (WB) test results.

METHODSFrom January 2004 to June 2009, three ELISA kits (Zhongshan, Biomérieux and Livzon) were used for initial screening HIV antibody. The reactive positive samples were reexamed by initial ELISA kit and a rapid kit (Abbot Determine HIV-1/2). All repeatedly reactive positive screening results were followed by WB test.

RESULTSA total of 193 (0.094%) WB confirmed positive results were obtained from 206 151 specimens. The sensitivities and predictive values of negative test result (PVN) of three ELISA kits were all 100% and those of Abbot Determine HIV-1/2 were 93.93%, and 91.67% respectively. All false negative results from Abbot were WB indeterminate. The specificities of Zhongshan, Biomérieux, Livzon and Abbot were 99.88%, 99.89%, 99.96% and 89.38%; the study predictive values of a positive test result (PVP) were 35.58%, 46.46%, 76.61% and 92.20%; the efficiencies were 99.88%, 99.89%, 99.96% and 91.98%; the areas under ROC curve of the three ELISA kits were 0.93, 0.99, and 0.95 respectively. PVP of Livzon was obviously higher than those of Zhongshan (chi(2) = 45.804, P = 0.000), Biomérieux (chi(2) = 25.231, P = 0.000) and Biomérieux was higher than Zhongshan (chi(2) = 2.488, P = 0.115). PVP of Abbot was highest (chi(2) = 18.633, P = 0.000, vs Livzon). There were some specimens with S/CO (optical density of sample/cut off) ratio < 6 or > or = 6 in all three groups with positive, indeterminate and negative WB results. The S/CO ratio from Zhongshan in confirmed positive group (14.29 + or - 2.63) was higher than in positive-negative group (2.80 + or - 3.25) (t = 17.652, P = 0.000). The S/CO ratio from Biomérieux in confirmed positive group(16.09 + or - 2.35) was higher than in positive-negative group (2.14 + or - 1.91) (t = 31.622, P = 0.000). The S/CO ratio from Livzon in confirmed positive group (11.54 + or - 1.95) was higher than in positive-indeterminate group (5.54 + or - 3.57) (t = 6.386, P = 0.000), positive-negative group (3.25 + or - 2.41) (t = 21.772, P = 0.000) and positive-indeterminate group was higher than positive-negative group (t = 2.301, P = 0.033).

CONCLUSIONThe performances of four HIV antibody screening kits are good but estimating WB confirming result in line with S/CO ratio is not available. All repeated screening positive results should be followed by confirmatory tests.

AIDS Serodiagnosis ; methods ; Blotting, Western ; methods ; Enzyme-Linked Immunosorbent Assay ; methods ; statistics & numerical data ; HIV Antibodies ; blood ; HIV Seropositivity ; diagnosis ; Humans ; Indicators and Reagents ; Mass Screening ; Reagent Kits, Diagnostic ; Sensitivity and Specificity

Notch signal path plays important roles in the regulation of proliferation and differentiation of hematopoietic stem cells. An extracellular domain of human Delta-like-1 (hDll-1(ext)), one of Notch ligands, was cloned and expressed in CHO cells, and the effect of hDll-1(ext) on expansion of hematopoietic stem/progenitor cells was investigated in this study. Total RNA was isolated from human marrow mononuclear cells. hDll-1(ext) was amplified by RT-PCR and cloned to T vector, then the gene was sequenced and subcloned to pcDNA3.1/Myc-His(+)A expression vector. The constructed plasmid was transfected into CHO cells with lipofectin and the expression of secreted hDll-1(ext) in G418-resistant clones was assayed by Western blot. hDll-1(ext) high-expressed clone was cultured to collect supernatant. Fusion protein hDll-1(ext) was purified from the supernatant by immobilized metal affinity chromatography (IMAC). The results showed that expression of Notch-1 receptor was detected in cord blood-derived CD34(+) cells by RT-PCR. Human umbilical blood CD34(+) cells were cultured in serum-free medium containing SCF, IL-3, VEGF, and with or without purified hDll-1(ext) for 4 or 8 days. Effect of hDll-1(ext) on the expansion of progenitor cells was analyzed then by clonogenic assays. The number of CFU-Mix and HPP-CFC generated from the culture system containing hDll-1(ext) was 1.5 times of that from the control. In conclusion, the recombinant hDll-1(ext) promotes the expansion of primitive hematopoietic progenitors.
Animals ; Antigens, CD34 ; immunology ; Binding Sites ; genetics ; CHO Cells ; Cell Division ; drug effects ; physiology ; Colony-Forming Units Assay ; Cricetinae ; Endothelial Growth Factors ; pharmacology ; Fetal Blood ; cytology ; immunology ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; Glycoproteins ; genetics ; pharmacology ; physiology ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Intercellular Signaling Peptides and Proteins ; pharmacology ; Interleukin-3 ; pharmacology ; Lymphokines ; pharmacology ; Membrane Proteins ; genetics ; RNA ; genetics ; metabolism ; Receptor, Notch1 ; Receptors, Cell Surface ; Recombinant Proteins ; isolation & purification ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cell Factor ; pharmacology ; Transcription Factors ; Transfection ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

BACKGROUNDNominal atrioventricular (AV) interval in dual chamber pacemaker (DDD) is not the best AV delay in the majority of patients with atrioventricular block. To find a simple method for optimizing AV delay adjustment, we assessed surface electrocardiography (ECG) for optimizing AV delay during dual chamber pacing.

METHODSDDD pacemakers were implanted in 46 patients with complete, or almost complete, AV block. Optimal AV delay was achieved by programming an additional delay of 100 ms, to the width of intrinsic P wave or to the interval between pacing spike to the end of P wave on surface ECG. Left ventricular (LV) end diastolic and end systolic volumes, ejection fraction and diastolic parameters were measured by Doppler echocardiography during both nominal and optimal AV delay pacing.

RESULTSCompared to nominal AV delay setting, LV end diastolic volume increased [to (53.2 +/- 11.3) ml from (50.2 +/- 10.2) ml, P < 0.05], end systolic volume decreased [to (26.1 +/- 9.0) ml from (27.9 +/- 8.2) ml, P < 0.05] during adjusted AV delay pacing, resulting in an increase in LV ejection fraction [to (68.2 +/- 5.3)% from (64.5 +/- 4.3)%, P < 0.05]. LV diastolic filling and isovolumic relaxation time were not significantly changed.

CONCLUSIONOptimization of AV delay by surface ECG is a simple method to improve LV systolic function during dual chamber pacing.

Adult ; Aged ; Aged, 80 and over ; Atrioventricular Node ; physiopathology ; Cardiac Pacing, Artificial ; methods ; Electrocardiography ; methods ; Female ; Heart Block ; physiopathology ; therapy ; Humans ; Male ; Middle Aged ; Time Factors