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.

Human osteosarcoma MG-63 cells were induced into differentiation by 5 mmol/L hexamethylene bisacetamide (HMBA). Their nuclear matrix proteins (NMPs) were selectively extracted and subjected to two-dimensional gel electrophoresis analysis.The results of protein patterns were analyzed by Melanie software. The spots of differentially expressed NMPs were excised and subjected to in situ digestion with trypsin. The maps of peptide mass fingerprinting were obtained by MALDI-TOFMS analysis, and were submitted for NCBI database searches by Mascot tool.There were twelve spots changed remarkably during the differentiation induced by HMBA, nine of which were identified. The roles of the regulated proteins during the MG-63 differentiation were analyzed. This study suggests that the induced differentiation of cancer cells is accompanied by the changes of NMPs, and confirms the presence of some specific NMPs related to the cancer cell proliferation and differentiation. The changed NMPs are potential markers for cancer diagnosis or targets for cancer therapy.

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.

Objective To observe the adverse reaction, tumor response and short term outcomes of percutaneous cryoablation for locally advanced pancreatic cancer, and investigate its feasibility. Methods Fifty-nine consecutive patients with locally advanced, unresectable pancreatic cancer underwent percutaneous cryoablation at our hospital from Sept. 2008 to Sept. 2009, were prospectively studied. Percutaneous cryoablation was performed with an argon/helium-based cryosurgical system under the guidance of ultrasound.Freezing probe was inserted into the center of pancreatic mass and two cycles of freezing were performed with each cycle for 5 min and temperature at-160℃, then the temperature was returned to normal for 10 min.Serum amylase was detected before operation and 1 to 7 days postoperatively. CT or PET-CT scanning was performed for evaluation of tumor response every 4 to 6 weeks after cryoablation. Survival was assessed by Kaplan-Meier method. Results 59 patients had a total of 76 biopsy-proven tumors, which were located at the pancreas head (n = 56), body (n = 7), and tail (n = 13). The median size of tumor was 4.5 cm (range 3 ～6 cm). Nineteen patients had liver metastases. Postoperative abdominal pain occurred in 45 cases (76.3%),fever occurred in 29 cases (49.2%) and elevation of serum amylase occurred in 34 cases (57.6%). Severe complications including intra-abdominal bleeding, pancreatic leaks, ileus, and metastasis by probe tract occurred in 5 cases (8.5%). There was no death associated with cryoablation. The median hospital stay was 21 days. 2 patients (3.4%) achieved complete response, 23 patients (39.0%) achieved partial response,30patients (50.8%) had stable disease, 4 patients(6.8%) had progressive disease. The median survival was 8.4 months. The overall survival at 3, 6 and 12 months was 89.7%, 61.1% and 34.5%, respectively.Conclusions Ultrasound-guided percutaneous cryoablation appears to be a safe and feasible, minimally invasive technique for locally advanced pancreatic cancer.

Objectives To investigate gestational multiple metabolic abnormalities aggregation and diagnostic criteria for gestational metabolic syndrome(GMS),and to analyze the risk factors of GMS.Methods A cohort study recruiting 309 pregnant women with preeclampsia,627 pregnant women with gestational diabetes mellitus(GDM)and 1245 normal pregnant women was performed from January 2008 to December 2011 in Guangdong Women and Children's Hospital.Information regarding age,gestational weeks,basic blood pressure,admission blood pressure,height and body mass index(BMI)before pregnancy was recorded.Biochemical indicators including fasting plasma glucose(FPG),fasting insulin (FINS),total cholesterol(TC),triglyceride(TG),high density lipoprotein(HDL-C),low density lipoprotein(LDL-C),free fatty acids(FFA)were tested.GMS was diagnosed with three or all of the following conditions:(1)overweight and/or obesity before pregnancy(BMI ≥ 25 kg/m2);(2)hypertension with blood pressure ≥ 140/90 mm Hg(1 mm Hg =0.133 kPa);(3)hyperglycemia:diagnosed as GDM;(4)dyslipidemia with TG≥3.23 mmol/L The incidence of GMS of the three groups were calculated and the risk factors were analyzed.Results(1)The age,gestational weeks,basic blood pressure,admission blood pressure,BMI before pregnancy of women with preeclampsia and women with GDM were significantly different compared to normal women,respectively(P ＜ 0.01).(2)Biochemical indicators of women with preeclampsia were as following:FPG(4.6 ± 1.0)mmol/L,FINS(10.1 ± 5.6)mU/L,TC(6.3 ±1.6)mmol/L,TG(3.9 ± 1.8)mmol/L,HDL-C(1.4 ±0.4)mmol/L,LDL-C(3.0 ± 1.0)mmol/L,FFA (0.8 ±0.4)mmol/L.And those in women with GDM were:FPG(4.7 ± 0.9)mmoL/L,FINS(10.2 ± 5.8)mU/L,TC(5.7 ± 1.3)mmol/L,TG(3.2 ± 1.1)mmol/L,HDL-C(1.4 ± 0.4)mmol/L,LDL-C (2.7 ± 0.9)mmol/L,FFA(0.6 ± 0.3)mmol/L In normal pregnant women they were:FPG(4.3 ±0.5)mmol/L,FINS(9.0±4.4)mU/L,TC(5.7 ±1.1)mmol/L,TG(2.8 ±1.1)mmol/L,HDL-C (1.5 ± 0.4)mmol/L,LDL-C(2.9 ± 0.8)mmol/L,FFA(0.6 ± 0.2)mmol/L Statistic differences were found in preeclampsia and GDM women compared to normal women respectively(P ＜ 0.01).(3)The prevalence of GMS in preeclampsia group and in GDM group was 26.2％(81/309)and 13.6％(85/627),statistically different from that of the control group(0)(P ＜0.01).(4)Compared to normal women,women with preeclampsia had higher risk of developing GMS(OR =1.62,95 ％ CI 1.31-2.00,P ＜ 0.01).The risk factors were BMI(OR =1.29,95％ CI 1.13-1.47)and TG(OR =2.49,95％ CI 1.87-3.31).Also,women with GDM had higher risk of developing GMS than normal women(OR =1.27,95％ CI 1.09-1.49,P ＜ 0.01),and the risk factors were BMI(OR =1.13,95 ％ CI 1.04-1.23)and TG(OR =1.16,95 ％ CI 1.02-1.33).TG was the independent risk factor in both preeclampsia women and GDM women(P ＜ 0.01,P ＜ 0.05).HDL-C seemed to have less importance in identifying GMS(P ＞ 0.05).Conclusions According to the GMS diagnostic criteria used in this study,some preeclampsia patients and some GDM women had aggregation of multiple metabolic abnormalities including pre-pregnancy overweight/obesity,hyperglycemia,high blood pressure and dyslipidemia.TG was the independent risk factor for GMS.HDL-C seemed to have less importance in identifying GMS.

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

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