Background Increase in ophthalmic optical medical instruments and microsurgical applications leads to retinal photochemical damage and other problems delivery of a variety of devices,so the in-depth study and understanding of its pathogenesis after retina light damage can provide a reference for the clinical treatment of related diseases.Objective This study was to investigate the therapeutic effect and relative mechanism of erythropoietin (EPO) on mouse retina photic injury by studying the expression of matrix metalloproteinases-2 (MMP-2)and MMP-9.Methods Fifty-two SPF BALB/c mice were randomized into normal control group,simple light-induced group and EPO pretreatment group by balloting method.The mice of simple light-induced group and EPO pretreatment group were continuously irradiated with 6000 lx diffuse light for 4 hours in a home-made box to establish the models of light-induced damage;while recombinant human EPO (rhEPO)of 5000 U/kg was intraperitoneally injected prior to the light exposure in the EPO pretreatment group.The expressions of MMP-2 and MMP-9 were examined at 6,12,36,72,96 hours and 7 days following light-exposure by immunohistochemistry.Results Edema and structural disorder of RPE cells appeared inthe simple light-induced group after light-exposure and aggravated with lapse of light-exposure time,but no similar change was seen until 7 days in the EPO pretreatment group.The immunohistochemistry findings showed that the expression of MMP-2(A value)in RPE cells was less in the normal mice.However,a large quantity of positive cells appeared in RPE layer 36 hours after light-exposure.Compared with the simple light-induced group,the positive expression of MMP-2 protein in EPO pretreatment group was significantly decreased,showing statistically significant differences among these three groups and different time points (Fgroup =3.68,P =0 04; Ftime =9.13,P=0.00).There was hardly any MMP-9 expression in the retina of the normal mice.In simple light-induced group,a few of positive cells appeared in RPE layer 6 hours after light-exposure and reached its peak 12 hours following light-exposure.The gradually down-regulation of MMP-9 expression happened 96 hours later following light-irradiation.The expression tendency of MMP-9 in EPO pretreatment group was similar to the simple light-induced group.Significant differences in expressions of MMP-9 were found among different groups and time points (Fgroup=3.61,P =0.04;Ftime =16.91,P=0.00).Conclusions MMP-2 and MMP-9 may be involved in the mechanism of retina photic injury by down-regulating the expression of MMP-2 and MMP-9 in RPE cells.

Trihydroxybenzoic acid dimmer is an anti-tumor compound which is separated from water-caltrop. This study is aimed to investigate its anti-proliferation effect on HL-60 cells and the possible mechanism of inducing apoptosis. MTT assay was used to test HL-60 cells proliferation. The apoptosis, ROS levels and the mitochondrial membrane potential were detected by flow cytometry. Cytochrome c released from mitochondria to cytosol fraction was detected by Western blotting. The activity of Caspase-9 and Caspase-3 were measured by colorimetric method. It was found to inhibit HL-60 cells proliferation in a dose and time-dependent manner. Compared with control group, it caused the increase of ROS levels and a concomitant dissipation of the mitochondrial membrane potential and induced cytosolic accumulation of cytochrome c and activities Caspase-9 and Caspase-3. Therefore it can be concluded that mitochondrial-dependent pathways was involved in its induction of apoptosis of HL-60 cells.
Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cytochromes c ; metabolism ; Fruit ; chemistry ; Gallic Acid ; isolation & purification ; pharmacology ; HL-60 Cells ; Humans ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondria ; metabolism ; physiology ; Plants, Medicinal ; chemistry ; Reactive Oxygen Species ; metabolism

Lu Dangshen, a traditional authentic medicinal material of Codonopsis Radix is mainly produced in Shangdang (Changzhi) area of Shanxi Province. Baitiao Dangshen is mainly produced in Gansu Province. Codonopsis Radix contains many kinds of components such as phenylpropanoids, polyalkynes, alkaloids, terpenes, fatty acids, flavonoids, and so on. At present, the effect of producing areas on its chemical compositions has not been systematically studied. This study analyzed the differences of metabolites among Codonopsis pilosula from different producing areas by UPLC-HRMS. PCA, OPLS-DA coupled with Thermo mzcloud online and local databases were used to compare the overall differences of metabolites among Codonopsis pilosula from different producing areas, and the chemical constituents were identified to further screen and find out the different metabolites and analyze the metabolic pathways by information retrieval in HMDB, PubChem, Chemspider and KEGG databases. The results showed that 72 differential metabolites were identified in this study. There were 15 kinds of up-regulated and 57 kinds of down-regulated metabolites of Lu Dangshen compared with Baitiao Dangshen. The top 30 metabolic pathways were analyzed by KEGG enrichment, and the most important metabolic pathways were phenylpropanoid biosynthesis, which was demonstrated that phenylpropanoid biosynthesis pathway and related intermediate metabolites could be used as the characteristics of distinguishing Lu Dangshen from different habitats of Codonopsis pilosula. The present study provided a basis for analyzing the influence of producing areas on the chemical components of Codonopsis pilosula and reasonably evaluating the quality of Codonopsis Radix, and also provided a new idea for expounding the authenticity of Lu Dangshen.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide ; toxicity ; Carcinogens ; toxicity ; Cell Line, Tumor ; DNA Damage ; drug effects ; Dose-Response Relationship, Drug ; HSP70 Heat-Shock Proteins ; biosynthesis ; Humans

Objective To evaluate the physiological and anatomic basis,indications,surgical skills, prevention of ureter injury and clinic outcomes of using high uterosacral ligament suspension(HUS)for correction of advanced uterine prolapse by the vaginal route.Methods Fifty women with advanced uterine prolapse underwent transvaginal HUS after vaginal hysterectomy with reconstruction of pubocervical and rectovaginal fascia to correct their uterine prolapse between June 2003 and September 2007.The average age of the women was 60.1 years.The mean follow-up period was 24 months(range 4-51 months).The degree of pelvic organ prolapse preoperatively and anatomic outcomes postoperatively were assessed with pelvic organ prolapse quantification system(POP-Q).Results The remnants of the uterosacral ligaments were clearly identified and palpated posterior and medial to the ischial spines by traction with a 24 cm long Allis clamp and used for successful vaginal vault suspension and reconstruction in all 50 consecutive advanced uterine prolapse patients.The ureter injury was avoided by complete knowledge of the ureter's course from the cervix/apex toward its insertion in the sacral region and how far outside of the uterosacral ligament,by uteri palpation and by suturing purposefully placed"deep"dorsally and posteriorly toward the sacrum,as well as by cystoscopy examination of the spillage of urine from both ureters.Mean POP-Q point C improved from 1.5 to-7.5 cm with a median follow-up of 24 months.If the successful HUS was defined as point C≤stage I prolapse,both the objective and subjective cure rates were as high as 100% with a maximum follow-up of 51 months.None of the 50 patients had repeat operation for recurrence of prolapse.There was no major intra-or postoperative complications,such as ureter and other pelvic organ injury.Conclusion HUS with fascial reconstruction seems to be a safe,minimal traumatic,tolerable and highly successful procedure for vaginal repair of advanced uterine prolapse.Because of the use of native tissue as suspension site HUS is more physiologic and cost effective.

OBJECTIVETo investigate the expression of cortical SCF/KIT system at different blood glucose level in mice.

METHODS27 male C57 mice were randomly divided into control group, diabetes group, and diabetes plus insulin group. The diabetic mice were induced by streptozotocin. Western-blot and double-immunofluorescence histochemistry were used to detect the expression of SCF and KIT.

RESULTSBoth methods indicate that the level of S-SCF and M-SCF were decreased significantly in the diabetes group, and this trend can be reversed effectively when the insulin was utilized.

CONCLUSIONThe decline of SCF might be one of underlying mechanisms of diabetic encephalopathy.

Animals ; Cerebral Cortex ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; Down-Regulation ; physiology ; Insulin ; therapeutic use ; Male ; Mice ; Mice, Inbred C57BL ; Proto-Oncogene Proteins c-kit ; metabolism ; Stem Cell Factor ; metabolism

BACKGROUNDTreating metastatic osteosarcoma has been challenged in past decades. Extracelluar matrix (ECM) proteins play an important role in the progression of osteosarcoma as they are pivotal components of the tumor microenvironment. Here, we identified potential genes belonging to the ECM and characterized the roles of these genes in the progression of osteosarcoma and their association with outcomes.

METHODSOsteosarcoma parental cell line MG63 and its derivative MG63-A1 with a high metastatic potential underwent oligonucleotide microarray analysis. Gene ontology analysis was used to screen deregulated genes between the 2 cell lines which were either upregulated or downregulated by more than 4 fold, particularly focusing on mRNAs encoding extracellular matrix proteins. The expression of resulting candidate genes was then validated by reverse transcription-PCR for mRNA expression as well as Western blotting for protein expression. Immunohistochemistry was performed on 37 osteosarcoma specimens to examine the potential role of the candidate genes in a clinical context.

RESULTSMicroarray data and gene ontology analysis showed that Tenascin-C, a critical component of the ECM, is significantly down-regulated in the highly metastatic cell line MG63-A1 compared with the parental osteosarcoma cell line MG63-wt. This finding was validated at mRNA and protein levels. Immunohistochemical analysis found that Tenascin-C is located in the intercellular space in osteosarcoma specimens. Furthermore, low-grade Tenascin-C expression (less than 20%) in osteosarcoma specimens was associated with poor survival.

CONCLUSIONSTenascin-C expression level correlates with the survival of osteosarcoma patients. Its biological functional role and underlying molecular mechanisms in the progression of osteosarcoma needs further investigation.

Adolescent ; Adult ; Bone Neoplasms ; metabolism ; mortality ; Cell Line, Tumor ; Child ; Female ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; Osteosarcoma ; metabolism ; mortality ; Prognosis ; RNA, Messenger ; analysis ; Tenascin ; analysis ; genetics

BACKGROUNDSingle-fiber electromyography (SFEMG) abnormality in the extensor digitorum communis (EDC) was reported in ocular myasthenia gravis (OMG), which indicated subclinical involvement beyond extraocular muscles in OMG patients. The relationship between the abnormal findings of SFEMG in EDC and the probability for OMG to develop generalized myasthenia gravis (GMG) is unknown. This retrospective study aimed to determine the predictive value of abnormality of SFEMG in EDC of OMG patients.

METHODSOne-hundred and two OMG patients underwent standard clinical diagnosis process and SFEMG test in EDC muscle when diagnosed and were clinically followed up for 5 years. The SFEMG data were compared between different clinical groups according to thymus status, onset age, and different outcome of OMG developing. Chances of progressing to GMG were compared between two different groups according to SFEMG and repetitive nerve stimulation (RNS) results, acetylcholine receptor antibody (AchRAb) titer, thymus status, and onset age.

RESULTSAbnormal SFEMG results were observed in 84 (82.4%) patients. The mean jitter, percentage of jitter >55 μs (%), and blocking were higher in OMG patients than in healthy volunteers. There were no statistical differences in jitter analysis between thymoma group and non-thymoma group (P = 0.65), or between the later OMG group and the later GMG group (P = 0.31), including mean jitter, percentage of jitter >55 μs (%), and blocking. Elderly group (≥45 years old) had a higher mean jitter than younger group (t = 2.235, P = 0.028). Total 55 OMG developed GMG, including 47 in abnormal SFEMG group while 8 in normal SFEMG group. There was no statistical difference in the conversion rates between the two groups (χ2 = 0.790, P = 0.140). RNS abnormality, AchRab titer, or onset age had no correlation with OMG prognosis (P = 0.150, 0.070, 0.120, respectively) while thymoma did (χ2 = 0.510, P = 0.020).

CONCLUSIONSFEMG test in the EDC showed high abnormality in OMG, suggesting subclinical involvement other than extraocular muscles. Nevertheless, the abnormal jitter analysis did not predict the prognosis of OMG according to clinical follow-up.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Electromyography ; methods ; Humans ; Middle Aged ; Myasthenia Gravis ; metabolism ; pathology ; physiopathology ; Prognosis ; Receptors, Cholinergic ; metabolism ; Retrospective Studies ; Young Adult

BACKGROUNDSingle-fiber electromyography (SFEMG) has been suggested as a quantitative method for supporting chronic partial denervation in amyotrophic lateral sclerosis (ALS) by the revised EI Escorial criteria. Although concentric needle (CN) electrodes have been used to assess jitter in myasthenia gravis patients and healthy controls, there are few reports using CN electrodes to assess motor unit instability and denervation in neurogenic diseases. The aim of this study was to determine whether quantitative changes in jitter and spike number using CN electrodes could be used for ALS studies.

METHODSTwenty-seven healthy controls and 23 ALS patients were studied using both CN and single-fiber needle (SFN) electrodes on the extensor digitorum communis muscle with an SFEMG program. The SFN-jitter and SFN-fiber density data were measured using SFN electrodes. The CN-jitter and spike number were measured using CN electrodes.

RESULTSThe mean CN-jitter was significantly increased in ALS patients (47.3 ± 17.0 μs) than in healthy controls (27.4 ± 3.3 μs) (P < 0.001). Besides, the mean spike number was significantly increased in ALS patients (2.5 ± 0.5) than in healthy controls (1.7 ± 0.3) (P < 0.001). The sensitivity and specificity in the diagnosis of ALS were 82.6% and 92.6% for CN-jitter (cut-off value: 32 μs), and 91.3% and 96.3% for the spike number (cut-off value: 2.0), respectively. There was no significant difference between the SFN-jitter and CN-jitter in ALS patients; meanwhile, there was no significant difference between the SFN-jitter and CN-jitter in healthy controls.

CONCLUSIONCN-jitter and spike number could be used to quantitatively evaluate changes due to denervation-reinnervation in ALS.

Amyotrophic Lateral Sclerosis ; physiopathology ; Electrodes ; Electromyography ; Humans ; Middle Aged ; Needles ; ROC Curve

OBJECTIVETo develop a quantitative real-time polymerase chain reaction (PCR) for detecting Bartonella henselae.

METHODSAccording to the 16S-23S rRNA intervening sequences (IVS) specific for B. henselae, one pair of primers and one TaqMan-MGB probe were designed. A quantitative real-time PCR was developed with the primers, the probe, and the IVS, a standard template, in DNA sequence detection system (ABI 7900HT).

RESULTSThe standard curve was established with the standard template and the relationship between the value of threshold cycle (Ct) and the DNA copy number was linear (r = 0.997). The sensitivity of this quantitative real-time PCR was about 1000 times higher than that of a common PCR used to detect homologous DNA. By this quantitative real-time PCR, the DNA sample of B. henselae was positively detected but not from other rickettsial or bacterial DNA samples. The variation coefficients of intra- and inter-assay reproducibility were 0.2%-1.9%. Using the real-time quantitative PCR to detect samples from mice that were experimentally infected with B. henselae, the small amount of B. henselae DNA was detected in blood samples on days 2, 3, and 5 and large amount of B. henselae DNA was detected in spleen samples on days 1 and 2 after infection.

CONCLUSIONResults from our study suggested that this quantitative real-time PCR was highly specific, sensitive and with good repeatability for detection of B. henselae. It seemed quite useful for rapid detection of tiny DNA of B. henselae in various samples and laboratory diagnosis of bartonellosis caused by B. henselae.

Animals ; Bartonella Infections ; diagnosis ; Bartonella henselae ; genetics ; DNA, Bacterial ; analysis ; Mice ; Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Sensitivity and Specificity