BackgroundThe effects of extremely low frequency electromagnetic fields (ELF-EMFs) on public health have attracted wide attentions.The association of the thermal effect of ELF-EMFs with cancer and ocular tissue damage has been of concern.However,the pathological changes of scleral tissue after exposure to ELF-EMFs as well as the relationship between these changes and myopia are still poorly understood.ObjectiveThe present study was to investigate the molecular pathological changes of human fetal scleral fibroblasts (HFSFs) after exposure to ELF-EMFs in vitro and to explore the possible mechanism in the occurrence and development of myopia.MethodsHFSFs were cultured and passaged and then exposed to 50 Hz electromagnetic fields,and HFSFs that did not receive the irradiation of ELF-EMFs were used as the control group.The expression of collagen type Ⅰ (COL1A1 ) mRNA and matrix metalloproteinase-2 (MMP-2) mRNA in cultured HFSFs were detected by real-time qualitative polymerase chain reaction (real-time PCR) under different magnetic field intensites (0,0.1,0.2,0.5,1.0 mT) and different exposure time (0,6,12,24,36,48 hours).Cell proliferation assay of HFSFs was detected by the cell counting kit 8 ( CCK8 ) assay.The expression levels of COL1 A1 and MMP-2 proteins in HFSFs were further confirmed by immunofluorescence staining.Results The expression of COL1A1 mRNA was significantly down-regulated under the exposure of 0.2 mT ELF-EMFs for 6 hours,in comparison with the control group;moreover,it decreased in parallel with the increased of flux density (0.099±0.008 vs.0.050±0.004) (P =0.009 ).The expression of MMP-2mRNA was up-regulated conspicuously after exposure to 0.1 mT ELF-EMFs for 24 hours,and it increased with exposure time in comparison with the control group ( 0.009 ±0.001 vs.0.018±0.003 ) ( P =0.038 ).Proliferation of HFSFs (A450) was inhibited following the exposure to 0.2 mT ELF-EMFs for 24 hours in comparison with the control group (P =0.009 ).The expression of COL1 A1 in the experimental group was decreased,compared with the control group,but the expression of MMP-2 was increased.ConclusionsELF-EMFs inhibit the proliferation of HFSFs and expression of COL1 A1 in HFSFs,which might be one of the reasons for the development of myopia.

BACKGROUND:Related studies have showed that poly D,L-lactide-co-glycolide can effectively package antisense oligonucleotides, smal interfering RNA, microRNA. Poly D,L-lactide-co-glycolide can better protect them against the destruction of the enzymes in vivo and have slow the drug release. Therefore, the number of drug administration can be reduced to achieve a long-term and effective therapeutic effect. ＜br> OBJECTIVE:To prepare poly D,L-lactide-co-glycolide-CXCR4-miRNA-nano-particles and to research the characteristics of the prepared nanoparticles. ＜br> METHODS:Poly D,L-lactide-co-glycolide-CXCR4-miRNA nanoparticles were prepared by double emulsion-evaporation process. Ultraviolet spectrophotometry was utilized for measurement of encapsulation efficiency and drug-loading rate, observing the shape of nanoparticles by transmission electron microscope, and measuring the size and distribution of nanoparticles by laser particle size analyzer. Sustained-release characteristics of nanoparticle suspension were observed in phosphate buffer. ＜br> RESULTS AND CONCLUSION:The prepared nanoparticles were spherical-shaped, smooth, evently distributed and inadhesive. The particle size was mainly distributed within 143-502 nm, with an average diameter of 280 nm. The average drug loading was (0.515±0.023)%, the average encapsulation ratio was 50.2%and difference between batches was smal . The nanoparticles could slowly release in vitro and the process initial y experienced the fast-release stage, and then reached a basical y stable platform stage at day 14. These finding indicate that the process to prepare poly D,L-lactide-co-glycolide CXCR4-miRNA-nanoparticles by double emulsion-evaporation is simple. The prepared nanoparticles are wel targeted and exhibit sustained-release effects.

OBJECTIVE: To study the effects of Wenyang Chubi Decoction (WYCBD), a compound traditional Chinese herbal medicine, on connective tissue growth factor (CTGF) and collagen-I (COL-I) in a mouse model of scleroderma. METHODS: Scleroderma was induced in BALB/c mice by daily local injection of bleomycin for three weeks and the mice were randomly divided into untreated, WYCBD-treated and normal saline (NS) treated groups, with another group of BALB/c mice as normal control. WYCBD and NS were given orally for one month respectively. Histopathology in the skin and lungs of the mice were examined. The CTGF and COL-I expressions in the skin or skin lesions were detected by immunohistochemical Elivision assay. RESULTS: The expression levels of CTGF and COL-I in the untreated group were significantly higher than those in the normal control group (P<0.05). Compared with the NS-treated group, the WYCBD-treated group had significant improvement in the skin and lung histopathology and remarkably decreased expression levels of CTGF and COL-I (P<0.05). CONCLUSION: Scleroderma mice showed high expressions of CTGF and COL-I in the skin. WYCBD had the effects of decreasing the CTGF and COL-I expressions and improving the skin fibrosis.

Objective To explore the gene expression pattern of sample of human ovarian carcimoma Method The difference in gene expression between normal and neoplastic human ovarian tissues were investigated,we described the assembly and utilization of a 512 member cDNA microarray Result Thirty seven genes expressed in ovarian cancer were screened out,14 genes were up regulated,23 genes were down regulated Conclusion cDNA microarray for analysis of gene expression pattern is an effective method to identify novel ovarian cancer associated genes

Objective To evaluate the effects of ELISPOT (enzyme-link immunospot) test using different samples in diagnosing tuberculous pleurisy. Methods Using T-Spot-TB kit to detect interferon-γlevel in pleural effusion and periph?eral blood from 164 patients with tuberculous pleural effusion and 102 patients without tuberculous pleural effusion. Number of spot forming cells (SFCs) as well as the specificity and sensitivity of the tests were compared between these two methods (ELISPOT using leural effusion or peripheral blood). Results The area under the ROC curve was 0.947 in pleural effusion Elispot test while it was 0.905 in peripheral blood Elispot test. The sensitivity of pleural effusion ELISPOT test in diagnosis of tuberculous pleurisy (95.1%) was significantly higher than that of peripheral blood ELISPOT test (89.0%). What’s more, the specificity of pleural effusion ELISPOT test in diagnosis of tuberculous pleurisy (90.2%) was higher than that in diagno?sis of peripheral blood ELISPOT test (88.2%). Conclusion The pleural effusion ELISPOT test is more valuable than periph?eral blood ELISPOT in the diagnosis of tuberculous pleuritis.

Child, Preschool ; Fatal Outcome ; Female ; Humans ; Respiratory Tract Infections ; complications ; Shock, Septic ; etiology ; Streptococcal Infections ; complications

This study aims to explore and analyze the condition of concurrent diseases and medicine use of traditional Chinese medicine (TCM) and western medicine among the patients with insomnia. One thousand and sxity seven cases of data from 20 national hospitals' hospital information system (HIS) databases were collected. The frequent concurrent diseases included hypertension (26.9%), brain blood supply insufficiency (24.93%), cerebral infarction (19.49%), blood lipoprotein disturbance (15.28%), coronary heart disease (14.15%), headache (10.68%), chronic gastritis (8.81%), type 2 diabetes mellitus (7.87%), depressive disorder (7.4%) and anxiety disorder (6.65%). The 10 most frequently-used western drugs included alprazolam (35.99%), aspirin (25.4%), olanzapine (24.18%), cinepazide (23.06%), flupentixol & melitracen (18.74%), zolpidem (18.37%), oxiracetam (15.65%), estazolam (15%), aniracetam (13.4%) and piracetam (13.31%). The 10 most frequently-used TCM included Shuxuening injection (16.4%), Shuxuetong injection (15.18%), extract of ginkgo biloba leaf (14.71%), gastrodin (12.46%), Dengzanxixin injection (11.34%), Xueshuantong (8.53%), Danhong injection (6.37%), compound liquorice tablet (5.81%), Sanqi Tongshu capsule (5.72%) and sowthistle-leaf ixeridium injection (5.34%). Among all combined uses, the most frequent western drug use was alprazolam and olanzapine, while combined use of hypnotic drug and Huoxuehuayu formula is the most frequent. This study concludes that the concurrent diseases mainly include cardio-cerebrovascular diseases, metabolic disorders and anxiety-depression disorders, with increasing tendency of diseases types by ages, especially for cardio-cerebrovascular diseases. The most frequently-used hypnotic is alprazolam in the insomnia patients, and it is worth being concerned about the off-label use of olanzapine as an antipsychotic for the treatment of insomnia However, due to the fact that all cases data are from the inpatients, these findings have some limitations.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Alprazolam ; therapeutic use ; Anti-Anxiety Agents ; therapeutic use ; Antipsychotic Agents ; therapeutic use ; Benzodiazepines ; therapeutic use ; Cerebral Infarction ; drug therapy ; epidemiology ; etiology ; Coronary Disease ; drug therapy ; epidemiology ; etiology ; Diabetes Mellitus, Type 2 ; drug therapy ; epidemiology ; etiology ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Headache ; drug therapy ; epidemiology ; etiology ; Humans ; Hypertension ; drug therapy ; epidemiology ; etiology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Sleep Initiation and Maintenance Disorders ; complications ; drug therapy ; epidemiology ; Young Adult

Objective To determine the role of interleukin-8 (IL-8) produced by tumor induced fibroblasts in the development of cutaneous melanoma. Methods B16 melanoma cells induced L929 fibroblasts phenotype was transdifferentiated to myofibroblasts (MF) by co-culture in vitro. MF was monitored by morphology and immunophenotype for a-SMA. The level of IL-8 was detected by ELISA. The effect on B16 cell proliferation rate was estimated using MIT method in vitro. Melanoma implanting model was constructed in C57 mice. Results L929 MF phenotype could be modulated by B16 melanoma cells-derived transforming growth factor-β1 (TGF-β1) and elevated the levels of IL-8. L929 MF did not influence the B16 melanoma cells viability in vitro, but shortened the time of tumor formation and increased the incidence rates of tumors in C57 implanting model mice. Conclusion Fibroblasts can be activated by tumor cells and produce IL-8, which acts as an inflammatory cytokine promoting the development of cutaneous melanoma.

Objective To develope a novel rabbit carotid body and carotid common artery model in vivo for the simulation of various intermittent hypoxia (IH) intensities, IH durations, IH reoxygenation (ROX) durations and continuous hypoxia (CH) modes.Methods Forty-five adult New Zealand rabbits (2.5～3.0 kg) were anesthetized while spontaneous breathing kept intact.The tissue surrounding the fight earetid common artery and carotid sinus nerve (CSN) were cleared and "single" chemoreceptor bundle of the CSN was revealed.Then suction electrodes were placed and CSN afferent activity was monitored and recorded carefully.The fight common carotid artery was exposed, cannulated to distal part and its proximal part was ligated.Preparations were challenged by changing the PO2 of the gas mixture equilibrating the perfusate.Alternatively perfusion (2 mL/min) of equilibrated porfusate bubbled with normoxia or hypoxia gas mixtures formed IH/ROX cycles in carotid common artery,simulating the pattern of hypoxic episodes seen in obstructive sleep apnea syndrome (OSAS), or with continuously perfusing hypoxia perfusate to form CH modes.All the perfusing procedures were regulated by a customized computer-controlled set and monitored using O2 gas analyzer.After the systematic exposures, carotid body, carotid common artery part distal to cannula,and carotid bifurcation were harvested as samples.Results The frequencies and average amplitudes of CSN chemoreceptor bundles afferent activities with normoxia peffusion were (0.17±0.03) impulse/s and (46.2±4.4) μV, and with hypoxia perfusion were (0.6±0.09) impulse/s and (87.4±6.6) μV, respectively.PO2 was (139±1.5) nun Hg in normoxia perfusate and (35.2±1.3) mm Hg in hypoxia perfusate.Conclusion This new carotid body and carotid common artery model is a valuable tool to study neurological and biochemical changes in various IH and CH modes.

Objective To identify the T cell epitopes from ovarian cancer associated anti-idiotypic antibody 6B11in order to explore the molecular basis of 6B11 induced cellular immune responses against ovarian cancer.Methods Potential human leukocyte antigen(HLA)A0201 ligands were predicted by using SYFPEITHI algorithm and tested by the T2 binding assay for screening of HLA-A2 binding peptides from 6B11 complimentary determining region(CDR).Cytotoxic T lymphocytes(CTL)to 6B11 or peptides were generated by 3 rounds of in vitro stimulation with 6B11 or peptide-pulsod dendritic cells(DC),and then tested by 51Cr-release assay to ascertain the CTL epitope of 6B11.Cell proliferation assay was performed by using 6B11 specific CTL as responder cells and peptide-pulsed DC as stimulators to ascertain the helper T lymphocyte(Th)epitope of 6B11.Cytokine assay and interferon-γ ELISPOT assay were performed to verify the CTL and Th epitope of 6B11 further.Results Light chain CDR3 peptide(VL CDR3)of 6B11 induced specific CTL to kill HLA-A2 and target antigen positive ovarian cancer cells,which could be blocked by anti human major histocompatibility complex(MHC)Ⅰ antibody.Heavy chain CDR3 peptide(VH CDR3)of 6B11 stimulated the proliferation of 6B11-primed CTL,which could be blocked mainly by anti human MHC-Ⅱ antibody,and further experiments showed that part of the VH CDR3 peptide-primed CTL killed K562 cells.Peptides in VL CDR3 and VH CDR3 were the CTL and Th epitope mimicking the original antigen of 6B11 respectively.Collaboration of 6B11 CTL and Th epitope,or 6B11 CTL epitope and keyhole limpet hemocyanin(KLH),the former was more powerful in inducing specific cellular immune responses against ovarian cancer.6B11 or corresponding CTL and Th epitope specific CIL secreted high levels of interleukin-2 (1630,1503 ng/L)and interferon-γ(5620,5421 ng/L),while basal level of interleukin-4 was detected (253,274 ng/L).ELISPOT assay confirmed the existence of specific interferon-γ secreting cells in 6811 or epitopes specific CTL(196/1×106 T cell,184/1×106 T cell).Conclusions VL CDR3 and VH CDR3 peptides of 6B11 are the CTL and Th epitopes mimicking original antigen which could duplicate the activity of intact 6B11 to induce the cellular immune responses against ovarian cancer.The results have significant theoretical and practical value in application of anti-idiotypic antibody ag anti tumor vaccine.The acquired CTL and Th epitopes as constituents of future multiple peptides against ovarian cancer could be used in peptide vaccine based ovarian cancer therapy.