Objectives:To investigate in vitro tumor killing effect of recombinant human perforin C terminal truncated 125 amino acid polypeptide (rhPFP C) and N terminal truncated 118 amino acid polypeptide (rhPFP N, 22 139aa) using liver cancer cell line SMMC 7721 as target cells. Methods:Recombinant human PFP C and PFP N peptide were expressed by E.coli in fusion form with glutathione S transferase(GST), and were purified by affinity chromatography with glutathione agarose. Liver cancer cell line SMMC 7721 was treated with fusion proteins in different concentrations for 24 h before surviving cells were measured with microscopy and colorimetric MTT assay. Results:After treatment with rhPFP C or rhPFP N, SMMC 7721 cell membrane damaged, appeared lysis. In MTT assay, optical density values in test group were significantly lower than those in control group. At concentration of 2.5 ?g/ml, the killing activity of rhPFP C and rhPFP N were 33.38% and 5.90% respectively. Conclusions:Both rhPFP C and rhPFP N showed obvious killing effect on liver cancer cell line SMMC 7721.The activity of rhPFP C was great higher than that of rhPFP N.

Background The aqueous flare is obvious in uveitic glaucomatous eye due to the damage of blood-aqueous barrier,especially following intraocular surgery.How to quantitatively determine the aqueous flare is important for us to understand the severity of inflammatory response.Objective This study was to assess inflammatory response following Ahmed glaucoma valve implantation in uveitic glaucomatous eye.Methods A nonrandomized controlled study was carried out under the approval of Ethic Committee of Zhongshan Ophthalmic Center of Sun Yat-sen University.Twenty-nine eyes of 29 subjects with refractory glaucoma were enrolled this hospital from October 2011 through July 2012.The patients were divided into the uveitic glaucoma group (10 eyes) and the other refractory glaucoma group (19 eyes) with the matched demography,and Ahmed glaucoma valve implantation was performed on all the eyes under the informed consent of each patient.The aqueous flare value was determined with FC-2000 flare-cell photometry and intraocular pressure (IOP) was measured with Goldmann tonometer before surgery and 1 day,3 days,1 week,2 weeks,1 month and 3 months after surgery.The outcomes were compared between the two groups,and the correlation of aqueous flare value with IOP was analyzed.Results In the uveitic glaucoma group,the aqueous flare values were 21.10 (10.50,38.58) photoparticles/ms,88.00 (23.55,168.63) photoparticles/ms and 29.90 (8.90,65.18)photoparticles/ms 1 day,3 days and 1 week after surgery,which were significantly higher than 13.53 (7.60,24.00) photoparticles/ms before surgery (all at P＜0.01).The same trend was found in the other refractory glaucoma group.In addition,the aqueous flare value was higher in the uveitic glaucoma group than that in the other refractory glaucoma group in 3 months after surgery (q=-3.445,P＜0.01).No significant differences were seen in IOP between the two groups at various time points (all at P＞0.05).Also,no significant correlations were seen between IOP and aqueous flare value in all patients at various time points (preoperation:rs =0.136,P =0.481; postoperative 1 day:rs =0.019,P =0.922 ; postoperative 3 days:rs =-0.035,P =0.858 ; postoperative 1 week:rs =0.317,P=0.094; postoperative 2 weeks:rs =0.034,P =0.861 ; postoperative 1 month:rs =-0.094,P =0.628 ; postoperative 3 months:rs =0.065,P =0.738).Conclusions FC-2000 flare-cell photometry can reflect the inflammatory reaction of the anterior chamber following Ahmed glaucoma valve implantation in various types of refractory glaucomatous eyes.The postoperative inflammatory response is more serious and lasting in uveitic glaucomatous eye.

Objective To explore the verification methods for the performance of quantitative detection kit of anti-dsDNA antibodiy with enzyme-linked immunosorbent assay (ELISA).Methods The precision was verified according to the EP15-A2 document approved by the American Clinical and Laboratory Standards Institute(CLSI).The accuracy was verified by detecting the samples of previous external quality evaluation(EQA),compared with the comparative kits and recovery test.The lower limit of detection(LLD) was calculated by the results of blank samples.The cut-off value was verified according to the C28-A3C document approved by CLSI and CNAS-CL39:Guidance on the Application of Accreditation Criteria for the Medical Laboratory Quality and Competence in the Field of Clinical Qualitative Immunology respectively.The improved Doumas method was used to verify the range of linearity.Results The measured intra-assay and inter-assay coefficients of variation were lower than those announced by the manufacturer or the calculated values according to the EP15-A2 document.The coincidence rates for negative and positive EQA samples between detected and expected values were 98.4％ (63/64) and 100％ (20/20) respectively.The total coincidence rate was 98.8％ (83/84).The coincidence rate for negative and positive samples between the results from candidate and comparative kits were 91.2％ (52/57) and 87.0％ (40/46) respectively.The total coincidence rate was 89.3％ (92/103) and the Kappa value was 0.783 (P =0.062),which implied excellent consistency between the two kits.The mean recovery rate was 99.65％.The measured LLD was 0.5 IU/mL which was lower than 1 IU/mL as claimed by the manufacturer.The measured cut-off value according to the CNAS-CL39 document was 18.51 IU/mL,which was close to 20 IU/mL announced by the manufacturer.Based on the C28-A3C method,the cut-off value could be approved.The linear regression equation was Y =0.978 8X-3.125 4,r2 =0.996 1.There was no statistical difference between the intercept (-3.125 4) and 0 (t =-0.772,P =0.483).The range of linearity was from 1.6 to 212.5 IU/mL,which was consistent with the values declared by the manufacturer.All the verifications of the five performances above-mentioned could be passed.Conclusion The precision,accuracy,LLD,cut-off value and range of linearity of the candidate quantitative ELISA kit for anti-dsDNA antibody were consistent with the statement of the manufacturer,which indicated the performance of the kits may meet the requirements of clinic diagnosis and treatment.A series of methods used in this study provided a simple protocol for verifying the performance of quantitative ELISA kits.

BACKGROUND: Coronary artery disease (CAD) risk in type Ⅱ diabetics is one of the hot topics in clinical research.OBJECTIVE: By using electron-beam tomography(EBT) technology, heart scans were performed to investigate coronary artery lesion in type Ⅱ diabetics and their CAD risks were evaluated as well.DESIGN: Comparative study based on type Ⅱ diabetics complicated with coronary artery lesion.SETTING: Gerontology department in a hospital affiliated to a military medical university of Chinese PLA.PARTICIPANTS: From January 2002 to June 2004, 93 inpatients between the ages of 36 and 78 in the Gerontology Department of Xijing Hospital, an affiliation to the Fourth Military Medical University of Chinese PLA, were selected(male 80, female 13) . Totally 25 of all the patients proved normal by physical examinations were set as normal control(male 22, female 3) group; other inpatients were divided into the abnormal blood glucose group and the coronary heart disease (CHD) group respectively according to their examination results. Of the 57 patients in the abnormal blood glucose group,49 were male and 8 were female. This group was divided into two subgroups:the impaired glucose tolerance group(IGT) containing 29 patients and the diabetes group containing 28 patients. In this group, 44 patients have had heart symptom of different extent. Of the 11 patients in the CHD group, 9 were male and 2 were female. Inclusion criteria: patients with type Ⅱ diabetes were defined by American Diabetes Association in 1979; patients with CHD were defined in the report of the Joint International Society and Federation of Cardiology/World Health Organization Task Force on the standardization of clinical nomenclature and criteria for diagnosis of ischemic heart disease in 1979, and they were all confirmed by coronary angiography. Exclusion criteeases.METHODS: Both plain EBT heart scanning and contrast-enhanced heart scanning were performed on the 93 patients to evaluate the occurrence of coronary artery calcification(CAC), the number of stenosed coronary arterie,and the number of soft plaques. Additionally, other examination results including blood glucose, blood HbA1c and blood lipid were also collected for comparison between the conditions of coronary artery and assessment of the CHD risk among groups.MAIN OUTCOME MEASUREMENTS: In all the groups, the occurrence of CAC, the number of stenosed coronary arteries and the number of plaques were compared, and the CHD risk was analyzed.RESULTS: The occurrence of coronary artery calcification was significantly increased in all the study groups as compared with the control group(x2 = 18.88, P < 0. 01 ). Increasing trend was also observed in the study groups when calculating the number of branches with coronary stenosis and plaques ( F = 10. 758, P < 0. 01; F = 9. 991, P < 0.01 ). CHD risk in diabetes was estimated to be 7. 514 (95% confidence interval, 1. 885 -63. 778).CONCLUSION: Coronary artery lesion can be easily observed in type Ⅱ diabetics,and high CHD risk is predicted in this population. Early diagnosis of CAD and early rehabilitation intervention in grade 2 are extremely important in improving the quality of life and reducing the disability ratio of type Ⅱ diabetics.Huang C, Gao JY, Wong XM, Li R, Yang AL, Zheng MW, Xue FB. Risk analysis of coronary artery disease in type Ⅱ diabetics.

Objective To assess the prevalence and risk of coronary artery disease (CAD) in Chinese adults with type 2 diabetes mellitus (T2DM) using electron beam computed tomography (EBCT) and EBCT angiography (EBCTA). Methods: Ninety-four cases were enrolled in this study including diabetes (n=28), impaired glucose tolerance (IGT, n=30), coronary heart disease (CHD, n=11), and control (n=25). Cardiac EBCT plain scanning and EBCTA were performed on all of these subjects to evaluate coronary artery calcification (CAC) scores, and number of segments of stenosed coronary arteries. Both CAC and/or coronary artery stenosis were defined as patients with coronary artery lesions (CAL). Results CAC scores were not different with the control, diabetes, IGT, or CHD (P＞0.05)groups. Compared to control (0.520±1.295), more stenosed coronary arteries segments (P＜0.05) were detected in diabetes (2.964±1.915), IGT (2.200±2.024), and CHD (2.273±1.679). Number of stenosed artery segments were correlated with age (r=0.215, P=0.019),postprandial glucose (r=0.224, P=0.015), total cholesterol (r=0.323, P=0.000), and duration of diabetes (r=0.208, P=0.004). The incidences of CAL in diabetes (96.43%), IGT (93.33%), and CHD (90.91%) was substantially higher than that in normal control (56.00%, P＜0.01).The odds ratio of CAL associated with having diabetes was estimated to be 7.514 (95% CI: 1.885-63.778). Conclusions Coronary artery lesions are prevalent in Chinese adults with type 2 diabetes, implying a high CAD risk. EBCTA holds potential in depicting the details of CAL and can be used to track the progression of CAD in diabetes patients.

The UPLC fingerprint of colistimethate sodium was established for the study of quality consistency.The chromatographic column was Acquity UPLC? Peptide CSH C18 (2.1 mm × 150 mm, 1.7 μm).The mobile phase A was phosphate buffer-acetonitrile (19∶1), and the mobile phase B was phosphate buffer-acetonitrile (1∶1).The mobile phase was in gradient elution at a flow rate of 0.3 mL/min.The column temperature was set at 30 °C and the detection wavelength was 210 nm.The similarity of the fingerprints was analyzed with the Similarity Evaluation System for Chromatographic Fingerprint of Tradition Chinese Medicine (Version 2012) in combination with content determination of multiple index components to evaluate the quality consistency of imported and domestic bulk drugs.The result showed that both the original and generic bulk drugs met the specified limit requirements in the European Pharmacopoeia standards, and that their UPLC fingerprints were highly similar, indicating that the quality of the two substances was consistent.Establishing a fingerprint for similarity evaluation and combining it with the results of indicator component content determination as a comprehensive evaluation method for the study of drug quality consistency of complex components has the characteristics of fast, accurate, and comprehensive, which is helpful for drug quality evaluation and provides ideas for the evaluation of antibiotic quality consistency of complex components.

A novel method was developed for the content assay and related substances determination of neomycin sulfate by high performance liquid chromatography combined with pulsed amperometric detection(HPLC-PAD). The HPLC was performed on Thermo AcclaimTMAmG C18(4. 6 mm×150 mm, 3 μm). The mobile phase consisted of aqueous solution with 2% trifluoroacetic acid containing 0. 01% pentafluoropropionic acid and 0. 6%NaOH. The pulsed amperometric detector was operated with aquadruple-potential waveform for the detection. Neomycin B, Neomycin C and thirteen related substances were adequately separated by the established HPLC conditions. The limits of detection(LOD)and quantification(LOQ)of neomycin B and neomycin C were both 1. 75 ng and 3. 5 ng, respectively. Good linearities of neomycin B and neomycin C were found in their respective ranges which their correlation coefficients were greater than 0. 998 5. The established method is characterized by high specificity, sensitivity and wide range of linearity which has a good application prospect and provides the basis for improving the standard and quality control of neomycin sulfate.

OBJECTIVES: This population-based, prospective cohort study investigated the association between glaucoma and mortality in older adults. METHODS: Participants aged 45 years or older at baseline (47.9% male) were enrolled in 2011 for the China Health and Retirement Longitudinal Study (CHARLS). All-cause mortality was observed during 7 years of follow-up. The baseline data were collected in the 2011 CHARLS, and participants were followed up for 7 years (until 2018). The risk of all-cause mortality was investigated using Cox proportional-hazards regression with age as the time scale, adjusting for significant risk factors and comorbid conditions. RESULTS: Among the 14,803 participants included, the risk of all-cause death was significantly higher among people with glaucoma than among those without glaucoma, after adjustment for other confounders (hazard ratio [HR], 1.46; 95% confidence interval [CI], 1.04 to 2.03). In a subgroup analysis based on the mean age of death, among those who were 75 years and older (n=1,231), the risk of all-cause death was significantly higher in patients with glaucoma than in those without glaucoma (HR, 1.89; 95% CI, 1.24 to 1.89). CONCLUSIONS Participants with glaucoma had a higher risk of all-cause mortality, especially those aged 75 years and above. Our findings revealed potential mechanisms underlying an association between glaucoma and all-cause mortality. They also highlighted the importance of glaucoma management to prevent premature death in middle-aged and older adults.

OBJECTIVES: This study evaluated the association between obesity and glaucoma in middle-aged and older people. A population-based retrospective cohort study was conducted using data from the China Health and Retirement Longitudinal Study. METHODS: Glaucoma was assessed via self-reports. Multivariate logistic regression analysis and a Cox proportional hazards model were used to assess the relationship between obesity and glaucoma risk. RESULTS: Older males living in urban areas who were single, smokers, and non-drinkers were found to have a significantly higher incidence of glaucoma (all p<0.05). Diabetes, hypertension, and kidney disease were also associated with higher glaucoma risk, while dyslipidemia was associated with lower risk (all p<0.05). After the model was adjusted for demographic, socioeconomic, and health-related variables, obesity was significantly associated with a 10.2% decrease in glaucoma risk according to the Cox proportional hazards model (hazard ratio, 0.90; 95% confidence interval [CI], 0.83 to 0.97) and an 11.8% risk reduction in the multivariate logistic regression analysis (odds ratio, 0.88; 95% CI, 0.80 to 0.97). A further subgroup analysis showed that obesity was associated with a reduced risk of glaucoma in people living in rural areas, in smokers, and in those with kidney disease (all p<0.05). Obesity also reduced glaucoma risk in people with diabetes, hypertension, or dyslipidemia more than in healthy controls (all p<0.05). CONCLUSIONS This cohort study suggests that obesity was associated with a reduced risk of glaucoma, especially in rural residents, smokers, and people with kidney disease. Obesity exerted a stronger protective effect in people with diabetes, hypertension, or dyslipidemia than in healthy people.

BACKGROUNDNeovascular glaucoma is a refractory disease, and difficult to manage. The aim of this study was to evaluate the clinical outcomes of Ahmed glaucoma valve implantation (AGVI) in neovascular glaucoma (NVG) and non-NVG patients.

METHODSThis prospective, non-randomized study included 55 eyes of 55 patients with refractory glaucoma; 27 had NVG (NVG group) and 28 had non-NVG (non-NVG group). All of the patients underwent AGVI. The NVG group was adjunctively injected with intravitreal ranibizumab/bevacizumab (IVR/IVB) before AGVI. Intraocular pressure (IOP) was the primary outcome measure in this study. Surgical success rate, number of antiglaucoma medications used, best corrected visual acuity (BCVA), and postoperative complications were analyzed between the groups.

RESULTSAll of the patients completed the study (follow-up of 12 months). Kaplan-Meier survival curve analysis indicated that the qualified success rates in the NVG and non-NVG groups at 12 months were 70.5% and 92.9%, respectively; this difference was significant (P = 0.036). The complete success rates in the NVG and non-NVG groups at 12 months were 66.7% and 89.3%, respectively (P = 0.049). Compared with preoperative examinations, the postoperative mean IOP and use of medications were significantly lower at all follow-up time points in both groups (all P < 0.05). There were significant differences in BCVA between the two groups at the 12-month follow-up (χ(2) = 9.86, P = 0.020). Cox proportional hazards regression showed NVG as a risk factor for surgical failure (RR = 15.08, P = 0.033). Postoperative complications were similar between the two groups.

CONCLUSIONSAGVI is a safe and effective procedure in refractory glaucoma, but the success rate of surgery was related to the type of refractory glaucoma. The complete and qualified success rates of NVG patient adjunctive anti-vascular endothelial growth factor treatment are still lower than those of non-NVG patients.

Adult ; Aged ; Female ; Glaucoma ; surgery ; Glaucoma Drainage Implants ; Glaucoma, Neovascular ; surgery ; Humans ; Male ; Middle Aged ; Proportional Hazards Models ; Prospective Studies