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Chia-Liang Tsai,1 Ching-Yun Huang,2 Yi-Ching Lu,1 Li-Mei Pai,3,4 Daniel Horák,5 Yunn-Hwa Ma1,6 1Department of Physiology and Pharmacology, Chang Gung University, Taoyuan, 33302, Taiwan, Republic of China; 2Institute of Biomedical Sciences, Chang Gung University, Taoyuan, 33302, Taiwan, Republic of China; 3Department of Biochemistry & Molecular Biology, College of Medicine, Chang Gung University, Taoyuan, 33302, Taiwan, Republic of China; 4Liver Research Center, Chang Gung Memorial Hospital, Taoyuan, 33305, Taiwan, Republic of China; 5Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Prague 6, 162 06, Czech Republic; 6Department of Medical Imaging and Intervention, Chang Gung Memorial Hospital, Taoyuan, 33305, Taiwan, Republic of ChinaCorrespondence: Yunn-Hwa Ma, Department of Physiology and Pharmacology, Chang Gung University, Guishan, Taoyuan, 33302, Taiwan, Republic of China, Email yhma@mail.cgu.edu.twBackground: Intravascular delivery of nanoparticles for theranostic application permits direct interaction of nanoparticles and vascular cells. Since vascular smooth muscle cells (VSMCs), the major components of the vascular wall, are constantly subjected to mechanical stimulation from hemodynamic influence, we asked whether cyclic strain may modulate internalization of magnetic nanoparticles (MNPs) by cultured VSMCs.Methods: Cyclic strain (1 Hz and 10%) was applied with Flexcell system in cultured VSMCs from rats, with cell-associated MNPs (MNPcell) determined by a colorimetric iron assay. Transmission and scanning electron microscopy were used for morphology studies. Confocal microscopy was used to demonstrate distribution of actin assembly in VSMCs.Results: Incubation of poly(acrylic acid) (PAA)-coated MNPs with VSMCs for 4 h induced microvilli formation and MNP internalization. Application of cyclic strain for 4– 12 h significantly reduced MNPcell by up to 65% (p < 0.05), which was associated with blunted microvilli and reduced vesicle size/cell, but not vesicle numbers/cell. Confocal microscopy demonstrated that both cyclic strain and fibronectin coating of the culture plate reduced internalized MNPs, which were co-localized with vinculin. Furthermore, cytochalasin D reduced MNPcell, suggesting a role of actin polymerization in MNP uptake by VSMCs; however, a myosin II ATPase inhibitor, blebbistatin, exhibited no effect. Cyclic strain also attenuated uptake of PAA-MNPs by LN-229 cells and uptake of poly-L-lysine-coated MNPs by VSMCs.Conclusion: In such a dynamic milieu, cyclic strain may impede cellular internalization of nanocarriers, which spares the nanocarriers and augments their delivery to the target site in the lumen of vessels or outside of the circulatory system.Graphical Abstract: Keywords: cyclic strain, magnetic nanoparticles, endocytosis, actin

Yau-Jiunn Lee,1 Chao-Ping Wang,2,3 Wei-Chin Hung,2,4 Wei-Hua Tang,1 Yu-Hung Chang,1 Der-Wei Hu,1 Yung-Chuan Lu,3,5 Teng-Hung Yu,2,4 Cheng-Ching Wu,2,4,6 Fu-Mei Chung,2 Chia-Chang Hsu4,7,8 1Lee’s Endocrinology Clinic, Pingtung 90000, Taiwan; 2Division of Cardiology, Department of Internal Medicine, E-Da Hospital, Kaohsiung 82445, Taiwan; 3School of Medicine for International Students, College of Medicine, I-Shou University, Kaohsiung 82445, Taiwan; 4The School of Chinese Medicine for Post Baccalaureate, College of Medicine, I-Shou University, Kaohsiung 82445, Taiwan; 5Division of Endocrinology and Metabolism, Department of Internal Medicine, E-Da Hospital, Kaohsiung 82445, Taiwan; 6Division of Cardiology, Department of Internal Medicine, E-Da Cancer Hospital, Kaohsiung 82445, Taiwan; 7Division of Gastroenterology and Hepatology, Department of Internal Medicine, E-Da Hospital, Kaohsiung 82445, Taiwan; 8Health Examination Center, E-Da Dachang Hospital, Kaohsiung 80794, TaiwanCorrespondence: Chia-Chang HsuDivision of Gastroenterology and Hepatology, Department of Internal Medicine, E-Da Hospital, Kaohsiung 82445, TaiwanTel +886-7-615-1100 ext. 5914 or 5018Email aladarhsu1107@gmail.comPurpose: This study aimed to investigate the common and unique risk factors and bidirectional relationship between chronic kidney disease (CKD) and nonalcoholic fatty liver disease (NAFLD) in patients with type 2 diabetes mellitus (T2DM).Patients and Methods: This was a cross-sectional study of patients with T2DM enrolled in a disease management program at two specialized diabetes outpatient clinics. Common and unique risk factors for CKD and NAFLD were examined using structural equation models (SEMs). SEMs were also used to examine direct and indirect effects of NAFLD on CKD and those of CKD on NAFLD.Results: A total of 1992 subjects with T2DM were enrolled in this study. In multivariate analysis, NAFLD was independently associated with the odds of CKD (adjusted odds ratio=1.59, 95% confidence interval=1.12– 2.25, P=0.009). SEMs showed that age, triglyceride, uric acid (UA), albumin, and HbA1c levels had statistically significant direct effects on CKD, and the final model could explain 22% of the variability in CKD. Age, triglycerides, body mass index (BMI), UA, white blood cell (WBC) count, serum glutamic pyruvic transaminase (SGPT) level, and smoking status had statistically significant direct effects on NAFLD, and the final model could explain 43% of the variability in NAFLD. The common risk factors contributing to both CKD and NAFLD were age, triglycerides, and UA. The unique risk factors were albumin and HbA1c for CKD, and BMI, WBC, SGPT, and smoking for NAFLD. In addition, SEM analysis also confirmed the bidirectional causal relationship between NAFLD and CKD.Conclusion: Common and unique risk factors and a bidirectional relationship existed between CKD and NAFLD in our patients with T2DM.Keywords: bidirectional relationship, chronic kidney disease, nonalcoholic fatty liver disease, risk factor, structural equation modeling, type 2 diabetes mellitus

Ting-Kuo Chang,1–3,* Yung-Chang Lu,1–3,* Shu-Ting Yeh,2,3 Tzu-Chiao Lin,2,3 Chun-Hsiung Huang,2–4 Chang-Hung Huang1–3,5 1Department of Medicine, MacKay Medical College, New Taipei City, Taiwan; 2Department of Orthopaedic Surgery, MacKay Memorial Hospital, Taipei, Taiwan; 3Department of Medical Research, MacKay Memorial Hospital, Taipei County, Taiwan; 4Department of Orthopaedic Surgery, Changhua Christian Hospital, Changhua, Taiwan; 5Department of Dentistry, National Yang-Ming University, Taipei, Taiwan*These authors contributed equally to this workCorrespondence: Chang-Hung HuangDepartment of Medical Research, MacKay Memorial Hospital, No. 45, Minsheng Road, Tamshui District, New Taipei City, TaiwanEmail changhung0812@gmail.comBackground: Graphene and its derivatives have recently gained popularity in the biomedical field. Previous studies have confirmed that both the mechanical strength and wear resistance of graphene-containing polyethylene have been greatly improved. Therefore, it is being considered as an alternative for artificial joint replacement liners. Based on the literature, the wear debris generated from the traditional polymers used for orthopedic liners could lead to particle-induced osteolysis and, consequently, failure of joint replacement. However, the biological response of this novel graphene-based polymer is still unclear. Therefore, the current study aimed to investigate the in vitro and in vivo biological effects of graphene and graphene oxide (GO) particles on bone.Materials and Methods: The biological responses of graphene and GO particles were tested via in vitro and murine calvarial in vivo models. In the in vitro model, murine macrophage cells were mixed with particles and hydrogel and printed into two differently designed scaffolds; the induced proinflammatory cytokines were then tested. In the murine in vivo model, the particle size distribution was measured via SEM, and these particles were then administrated in the calvarial area, referring to our established model. A micro-CT and histological analysis were performed to examine the biological effects of the particles on bone health. The data were analyzed via the one-way analysis of variance to determine the differences between the groups.Results: Both graphene and GO induced significantly higher TNF-α and IL-6 secretion compared with the control in the three-dimensional in vitro model. In the murine calvarial in vivo test, the graphene and GO particles increased the bone mass compared with the sham groups in the micro-CT analysis. Bone formation was also observed in the histological analysis.Conclusion: In these in vivo and in vitro studies, the graphene and GO wear debris did not seem to induce harmful biological response effect to bone. Bone formation around the skull was observed in the calvarial model instead. Graphene-containing biomaterials could be a suitable new material for application in orthopedic prostheses due to their benefit of eliminating the risk of particle-induce osteolysis.Keywords: biological response, graphene, graphene oxide, osteolysis, osteogenesis

Chih-Cheng Lai,1 Chi-Chung Chen,2,3 Ying-Chen Lu,3 Hung-Jui Chen,4 Bo-An Su,4 Tzu-Chieh Weng,4 Yu-Hsin Chiu,5 Han-Siong Toh,4 Chun-Cheng Zhang,4 Shyh-Ren Chiang,4 Yin-Ching Chuang,2,5 Hung-Jen Tang2,4,6 1Department of Intensive Care Medicine, Chi Mei Medical Center, Liouying, Tainan, Taiwan; 2Department of Medical Research, Chi Mei Medical Center, Tainan, Taiwan; 3Department of Food Science, National Chiayi University, Chiayi, Taiwan; 4Department of Medicine, Chi Mei Medical Center, Tainan, Taiwan; 5Department of Internal Medicine, Chi Mei Medical Center, Liouying, Tainan, Taiwan; 6Department of Health and Nutrition, Chia Nan University of Pharmacy and Science, Tainan, Taiwan Abstract: New-Delhi metallo-β-lactamase1 (NDM-1) Enterobacteriaceae are increasing worldwide. Herein, we describe a single patient who carried three unusual NDM-1 carbapenem-resistant Enterobacteriaceae – Enterobacter cloacae (E. cloacae) yielded from a urine specimen and Klebsiella pneumoniae (K. pneumoniae) and Escherichia coli (E. coli) from stool specimens. For E. cloacae, its bla NDM-1-encoding plasmid was pKP04NDM with a size of ~54 kb replicons with an IncN backbone. For K. pneumoniae, its bla NDM-1-encoding plasmid was pNDM-BTR with a size of ~59.6 kb and belonged to IncN. For E. coli, its main bla NDM-1-encoding plasmid was pIMP-HK1500, and the NDM-1 gene was obtained from a part of pNDM-BTR (8439 bp). These three clinical strains are reported for the first time and are assumed to be imported from mainland China to Taiwan. The three different plasmids were never reported in K. pneumoniae, E. coli, and Citrobacter spp before. Owing to their associated multidrug resistance, appropriate measures of periodic, targeted surveillance, and development of new antimicrobial agents are urgently needed.Keywords: carbapenem-resistant Enterobacteriaceae, NDM-1, K. pneumoniae, E. coli, Citrobacter, plasmid

Chih-Cheng Lai,1 Chi-Chung Chen,2,3 Ying-Chen Lu,3 Yin-Ching Chuang,2,4 Hung-Jen Tang5,6 1Department of Intensive Care Medicine, Chi Mei Medical Center, Liouying, Tainan, Taiwan; 2Department of Medical Research, Chi Mei Medical Center, Tainan, Taiwan; 3Department of Food Science, National Chiayi University, Chiayi, Taiwan; 4Department of Internal Medicine, Chi Mei Medical Center, Liouying, Tainan, Taiwan; 5Department of Medicine, Chi Mei Medical Center, Tainan, Taiwan; 6Department of Health and Nutrition, Chia-Nan University of Pharmacy and Science, Tainan, Taiwan Background: This study aimed to investigate the in vitro activity of cefoperazone–sulbactam against carbapenem-resistant Acinetobacter baumannii and Pseudomonas aeruginosa, and to evaluate the antibiotic resistance mechanisms of these bacteria. Materials and methods: In total, 21 isolates of carbapenem-resistant P. aeruginosa and 15 isolates of carbapenem-resistant A. baumannii with different pulsed-field gel electrophoresis types were collected for assessment of the in vitro antibacterial activities of cefoperazone and cefoperazone–sulbactam and the associated resistance mechanisms of the bacteria. Results: For carbapenem-resistant P. aeruginosa, the minimum inhibitory concentration (MIC) value and antibiotic susceptibility rate were similar for cefoperazone and cefoperazone–sulbactam (at 1:1 and 2:1 ratios). In contrast, for carbapenem-resistant A. baumannii, the MIC values, including the MIC range, MIC that inhibited 50% of isolates (MIC50) and MIC that inhibited 90% of isolates (MIC90), were reduced after treatment with sulbactam and cefoperazone. We screened resistance genes, including VIM-2, OXA-2 and OXA-10, in 21 carbapenem-resistant P. aeruginosa isolates. Only one (4.8%) of the isolates showed expression of VIM-2, and neither the OXA-2 nor the OXA-10 gene was detected. However, 20 (95.2%) isolates among the carbapenem-resistant P. aeruginosa isolates selected for oprD sequencing showed the phenomenon of nucleotide substitution or deletion. Among 15 carbapenem-resistant A. baumannii isolates, we found that ten (66.7%) isolates had concomitant expression of the OXA-23 and ISAba1-OXA-23 genes, and six (40.0%) isolates had expression of the OXA-24-like gene. All 15 isolates had OXA-51-like gene expression, and only 1 (6.7%) isolate had ISAba1-OXA-51-like gene expression. None of the isolates contained the IMP-1, IMP-8, KPC, NDM, VIM-1 or OXA-48 genes. Conclusion: The in vitro antibacterial activity of cefoperazone against carbapenem-resistant A. baumannii can be enhanced by adding sulbactam to cefoperazone, but the addition does not affect carbapenem-resistant P. aeruginosa. This significant difference can be explained by the different resistance mechanisms of carbapenem-resistant A. baumannii and P. aeruginosa. Keywords: cefoperazone–sulbactam, Acinetobacter baumannii, Pseudomonas aeruginosa

Chih-Cheng Lai,1 Chi-Chung Chen,2,3 Ying-Chen Lu,3 Tsuey-Pin Lin,4 Yin-Ching Chuang,2,5 Hung-Jen Tang4,6 1Department of Intensive Care Medicine, Chi Mei Medical Center, Liouying, 2Department of Medical Research, Chi Mei Medical Center, Tainan, 3Department of Food Science, National Chiayi University, Chiayi, 4Department of Health and Nutrition, Chia-Nan University of Pharmacy and Science, 5Department of Internal Medicine, Chi Mei Medical Center, Liouying, 6Department of Medicine, Chi Mei Medical Center, Tainan, Taiwan Objectives: This study aims to assess the in vitro activity of different cefoperazone–sulbactam ratios against different multidrug-resistant organisms (MDROs).Materials and methods: Minimum inhibitory concentrations (MICs) and susceptibility rates of cefoperazone, sulbactam and cefoperazone–sulbactam at fixed ratios of 2:1, 1:1 and 1:2 against 344 MDRO clinical isolates, including extended-spectrum β-lactamase (ESBL)-producing Escherichia coli (n=58), ESBL-producing Klebsiella pneumoniae (n=58), carbapenem-resistant Enterobacteriaceae (n=57), carbapenem-resistant Pseudomonas aeruginosa (n=49) and carbapenem-resistant Acinetobacter baumannii (n=122), were measured.Results: Combined treatment with sulbactam and cefoperazone resulted in decreased MIC50 values across all MDROs, as well as decreases in most MIC90 values, except for carbapenem-resistant Enterobacteriaceae and carbapenem-resistant P. aeruginosa (MIC90 values remained >64 mg/L). Susceptibility rates of treatment with cefoperazone alone against all MDROs were much lower than that of cefoperazone–sulbactam combination (all P

Yi-Ping Fang,1,2 Chih-Hung Chuang,3 Yi-Jhun Wu,1 Hsin-Che Lin,1 Yun-Chi Lu4 1School of Pharmacy, College of Pharmacy, Kaohsiung Medical University, 2Department of Medical Research, Kaohsiung Medical University Hospital, 3Department of Medical Laboratory Science and Biotechnology, College of Health Sciences, Kaohsiung Medical University, 4Graduate Institute of Medicine, Collage of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan Background: SN38 (7-ethyl-10-hydroxycamptothecin) is a camptothecin derivative acts against various tumors. However, SN38 is hydrolyzed in the physiological environment (pH 7.4), and this instability interferes with its potential therapeutic effect. Our objective was to investigate SN38-loaded liposomes to overcome the poor solubility of SN38 and its biodistribution, which further diminish its toxicity.Materials and methods: The sub-100 nm targeted liposomes was employed to deliver SN-38 and evaluate the characterization, release behaviors, cytotoxicity, in vivo pharmacokinetics and biochemical assay.Results: The SN38-loaded targeted liposomes consisted of small (100.49 nm) spherical nanoparticles with negative charge (-37.93 mV) and high entrapment efficiency (92.47%). The release behavior of the SN38-loaded targeted liposomes was fitted with Higuchi kinetics (R2=0.9860). Free SN38 presented initial burst release. The IC50 for the SN38-loaded targeted liposomes (0.11 µM) was significantly lower than for the SN38 solution (0.37 µM) in the MCF7 cell line (P

Chih-Cheng Lai,1 Chi-Chung Chen,2,3 Ying-Chen Lu,3 Yin-Ching Chuang,2,4 Hung-Jen Tang5,6 1Department of Intensive Care Medicine, Chi Mei Medical Center, Liouying, Tainan, Taiwan; 2Department of Medical Research, Chi Mei Medical Center, Tainan, Taiwan; 3Department of Food Science, National Chiayi University, Chiayi, Taiwan; 4Department of Internal Medicine, Chi Mei Medical Center, Liouying, Tainan, Taiwan; 5Department of Health and Nutrition, Chia-Nan University of Pharmacy and Science, Tainan, Taiwan; 6Department of Medicine, Chi Mei Medical Center, Tainan, Taiwan Background: The aim of this study was to investigate the genotypic differences between different sequence type MRSA isolates, especially focusing on silent rpoB474 mutations and the relationship between such mutations and ciprofloxacin resistance. Methods: Seventy-nine MRSA isolates were obtained for antibiotic susceptibility tests and molecular study. Results: Among these isolates, we found that the MIC50, MIC90, and minimum inhibitory concentration (MIC) range of ciprofloxacin were much higher for the isolates without the rpoB474 mutation than for isolates with the rpoB474 mutation. A total of 87.5% of the isolates with the rpoB474 mutation were susceptible to ciprofloxacin, but none of the isolates without the rpoB474 mutation were susceptible to ciprofloxacin. For 27 MRSA isolates without rpo474 silent mutation but with gyrA86/126 silent mutation, all of them belonged to SCCmec III, and had high ciprofloxacin MIC levels. For another 44 MRSA isolates with rpo474 silent mutation but without gyrA86/126 silent mutation, all of them showed low ciprofloxacin MIC levels, all of them belonged to either SCCmec IV or V. Furthermore, MRSA ciprofloxacin resistance was found to be associated with the mutations gyrA S84L/parC S80F or gyrA S84L, and S85P/parC S80Y. Conclusion: Most occurrences of this rpoB474 silent mutation were found in community acquired-MRSA (CA-MRSA) isolates with susceptibility to most antibiotics, especially for ciprofloxacin and vice versa. Thus, this mutation may help to differentiate the different microbiologic characteristics of MRSA clinical isolates. Keywords: MRSA, silent mutation, rpoB474, ciprofloxacin, resistance

Wei Xiong Siow,1,2 Yi-Ting Chang,1,2 Michal Babič,3 Yi-Ching Lu,2 Daniel Horák,3 Yunn-Hwa Ma1,2,4 1Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Guishan, Taoyuan, Taiwan, Republic of China; 2Department of Physiology and Pharmacology and Healthy Aging Research Center, College of Medicine, Chang Gung University, Guishan, Taoyuan, Taiwan, Republic of China; 3Institute of Macromolecular Chemistry, Czech Academy of Sciences, Prague, Czech Republic; 4Department of Neurology, Chang Gung Memorial Hospital, Guishan, Taoyuan, Taiwan, Republic of China Background: Poly-L-lysine (PLL) enhances nanoparticle (NP) uptake, but the molecular mechanism remains unresolved. We asked whether PLL may interact with negatively charged glycoconjugates on the cell surface and facilitate uptake of magnetic NPs (MNPs) by tumor cells.Methods: PLL-coated MNPs (PLL-MNPs) with positive and negative ζ-potential were prepared and characterized. Confocal and transmission electron microscopy was used to analyze cellular internalization of MNPs. A colorimetric iron assay was used to quantitate cell-associated MNPs (MNPcell).Results: Coadministration of PLL and dextran-coated MNPs in culture enhanced cellular internalization of MNPs, with increased vesicle size and numbers/cell. MNPcell was increased by eight- to 12-fold in response to PLL in a concentration-dependent manner in human glioma and HeLa cells. However, the application of a magnetic field attenuated PLL-induced increase in MNPcell. PLL-coating increased MNPcell regardless of ζ-potential of PLL-MNPs, whereas magnetic force did not enhance MNPcell. In contrast, epigallocatechin gallate and magnetic force synergistically enhanced PLL-MNP uptake. In addition, heparin, but not sialic acid, greatly reduced the enhancement effects of PLL; however, removal of heparan sulfate from heparan sulfate proteoglycans of the cell surface by heparinase III significantly reduced MNPcell.Conclusion: Our results suggest that PLL-heparan sulfate proteoglycan interaction may be the first step mediating PLL-MNP internalization by tumor cells. Given these results, PLL may facilitate NP interaction with tumor cells via a molecular mechanism shared by infection machinery of certain viruses. Keywords: magnetic nanoparticles, poly-L-lysine, tea catechin, glycoconjugate, heparan sulfate proteoglycan

Hung-Wei Yang,1,* Mu-Yi Hua,1,* Kun-Ju Lin,2,* Shiaw-Pyng Wey,3 Rung-Ywan Tsai,4 Siao-Yun Wu,5 Yi-Ching Lu,5 Hao-Li Liu,6 Tony Wu,7 Yunn-Hwa Ma5 1Chang Gung Molecular Medicine Research Center, Department of Chemical and Materials Engineering, 2Molecular Imaging Center, Department of Nuclear Medicine, Chang Gung Memorial Hospital, Kuei-Shan, Tao-Yuan, Taiwan, Republic of China; 3Department of Medical Imaging and Radiological Sciences, 4Electronics and Optoelectronics Research Laboratories, Industrial Technology Research Institute, Hsin-chu, Taiwan, Republic of China; 5Department of Physiology and Pharmacology and Healthy Aging Research Center, 6Department of Electrical Engineering, Chang Gung University, Kuei-Shan, Tao-Yuan, Taiwan, Republic of China; 7Department of Neurology, Chang Gung University College of Medicine and Memorial Hospital, Tao-Yuan, Taiwan, Republic of China*These authors contributed equally to this workAbstract: Low-toxicity magnetic nanocarriers (MNCs) composed of a shell of poly [aniline-co-N-(1-one-butyric acid) aniline] over a Fe3O4 magnetic nanoparticle core were developed to carry recombinant tissue plasminogen activator (rtPA) in MNC-rtPA for targeted thrombolysis. With an average diameter of 14.8 nm, the MNCs exerted superparamagnetic properties. Up to 276 µg of active rtPA was immobilized per mg of MNCs, and the stability of the immobilized rtPA was greatly improved during storage at 4°C and 25°C. In vitro thrombolysis testing with a tubing system demonstrated that magnet-guided MNC-rtPA showed significantly improved thrombolysis compared with free rtPA and reduced the clot lysis time from 39.2 ± 3.2 minutes to 10.8 ± 4.2 minutes. In addition, magnet-guided MNC-rtPA at 20% of the regular rtPA dose restored blood flow within 15–25 minutes of treatment in a rat embolism model without triggering hematological toxicity. In conclusion, this improved system is based on magnetic targeting accelerated thrombolysis and is potentially amenable to therapeutic applications in thromboembolic diseases.Keywords: thrombolysis, recombinant tissue plasminogen activator, magnetic nanocarriers, magnetic targeting, targeting therapy

Coastal wetland, at the intersection of land and sea, is considered as a "sink", "source", and "transformer" of phosphorus (P). Coastal wetland plays an important role in the global P cycle, and its ability to retain excessive P in water receives increasing attention. In this study, the coastal reed wetland sediments surrounding the Bohai Sea were sampled to investigate P adsorption capacity and loss risk by conducting batch experiments. Results show that the maximum P adsorption capacity ( Q max ) was 693.7-2117.2 mg·kg -1 , with an average of 1468.6 mg·kg -1 . The Q max decreased in the order of Qilihai Wetland > Beidagang Wetland > Nandagang Wetland > Liaohe Delta Wetland > Shouguang Coastal Wetland > Yellow River Delta Wetland. The P adsorption capacity was related to the contents of Ca, Mg, and TOC. The degree of P adsorption saturation (DPS) and loss risk index (ERI) of the coastal wetland were 0.28%-4.50% and 0.53%-10.10%, respectively. The ERI suggested that the P loss risk was relatively low for coastal reed wetland surrounding the Bohai Sea except for the moderate loss risk for Shouguang coastal wetland. In summary, the reed coastal wetland around Bohai Sea demonstrated significantly P storage capacity and served as a P sink for water P. We recommend to fully utilize the coastal wetland to reduce point or non-point source pollution ( e.g., P) during the remediation or recovery of the polluted Bohai Sea.

Objective: To investigate the perfusion characteristics of arterial spin labeling (ASL) in intracranial tumor and its application value in classification. Methods: The clinical, pathological and imaging data of 44 patients with gliomas confirmed by pathology were analyzed retrospectively, including 9 low grade gliomas, 15 high grade gliomas, 11 cases of meningiomas, 6 cases of neurilemmoma, 3 cases of metastatic tumors.Conventional plain scan, 3D- ASL and MRI dynamic enhanced imaging (DSC-MRI) were performed.The mean maximal cerebral blood flow (CBF) of the solid component of tumor was obtained based on the region of interest.Immunohistochemical staining was performed in 24 patients with glioma.The differences of cerebral blood flow map (CBF) and relative cerebral blood flow (rCBF) in 44 patients with intracranial tumors were compared. The results of paired t test between the tumor area and the contralateral mirror area were measured by the two methods. Results: Taken the normal control-lateral grey matter(GM) as reference to normalize the CBF of tumor, three normalized tumor blood flow (nTBF) acquired by ASL showed statistical difference between low grade and high grade gliomas respectively ( P <0.05). While taken the mirror region (M) and normal control-lateral white matter (WM) as reference to normalize the CBF of tumor, it showed no statistical difference ( P >0.05). There was no 1p deletion in the cases of ASL perfusion in low-grade glioma group.In the case of 1p deletion in high grade glioma group, ASL was low perfusion, and there was no 1p deletion in the cases of ASL perfusion. Conclusion: 3D ASL can be used to identify high-grade and low-grade gliomas which has important reference value in the qualitative diagnosis of brain tumors and preoperative grading of gliomas.A separate use of 3D-ASL might cause over-or underestimation of tumor diagnosis, therefore a comprehensive analysis is needed.

Estuary is an important area contributing to the global carbon cycle. In order to analyze the spatial-temporal distribution characteristics of the dissolved inorganic carbon (DIC) in the surface water of Yellow River estuary. Samples were collected in spring, summer, fall, winter of 2013, and discussed the correlation between the content of DIC and environmental factors. The results show that, the DIC concentration of the surface water in Yellow River estuary is in a range of 26.34-39.43 mg x L(-1), and the DIC concentration in freshwater side is higher than that in the sea side. In some areas where the salinity is less than 15 per thousand, the DIC concentration appears significant losses-the maximum loss is 20.46%. Seasonal distribution of performance in descending order is spring, fall, winter, summer. Through principal component analysis, it shows that water temperature, suspended solids, salinity and chlorophyll a are the main factors affecting the variation of the DIC concentration in surface water, their contribution rate is as high as 83% , and alkalinity, pH, dissolved organic carbon, dissolved oxygen and other factors can not be ignored. The loss of DIC in the low area is due to the calcium carbonate sedimentation. DIC presents a gradually increasing trend, which is mainly due to the effects of water retention time, temperature, outside input and environmental conditions.

An optical monitoring method is proposed for the rapid, non destructive measurements of chlorophyll concentration (Chl-a) in the surface sediments of emerged tidal flat, and it can be further applied in remote sensing work. Hyperspectral reflectance of intertidal sediments were measured in day time at the tidal flats of the Sishili Bay, the Northern Yellow Sea, and surface sediments (3 mm) were sampled for the in-door measurements of Chl-a. On the basis of the reflectance at 650, 675 and 700 nm, the indices of normalized difference index of microbenthos (NDI-MPB) and trough depth (T-depth) were proposed for the measurements of microphytobenthos biomass. T-depth can be used to remove the linear background spectral noises and indicate the existence of microphytobenthos; Good linear relationship was observed between NDI-MPB and Chl-a content in sediments (2.22-49.36 mg x m(-2), r > 0.99), which may be used to monitor the biomass of microphy to benthos.

Objective: To evaluate the clinical efficacies of restorative proctocolectomy with ileal pouch-anal anastomosis (RP-IPAA) for ulcerative colitis (UC)., Methods: A total of 115 UC patients underwent the procedure of RP-IPAA during the period of 1989 to 2010. The data of surgical procedures, complications and long-term outcomes were collected. Stool frequency during the 24-hour period and the night were recorded at months 3 and 12 post-operation. Anal continence functions were evaluated by the Kirwan grading scale and the fecal characteristics identified by Bristol stool form scale. The patients' quality of life was objectively accessed by the Cleveland Global Quality of Life (CGQL) index., Results: No perioperative death was reported. The mean follow-up period was for at least 12 months. The postoperative complication rate was 20.9% (24/115), including infectious complications, wound infection and(or) abdominal abscess, anastomotic leak, pouch hemorrhage, pouch-vaginal fistula, pouchitis, terminal proctitis, urinary calculi and male sexual dysfunctions. Stool frequency per 24 hours and night was < 3 to 12 month post-operation (3.9 ± 1.7 vs 6.1 ± 2.6, 1.3 ± 0.7 vs 2.8 ± 1.8, both P < 0.05). No significant changes in anal continence functions existed between the above two time points [Grade I: 87.0% (n = 100) vs 92.2% (n = 106), Grade II: 7.8% (n = 9) vs 4.3% (n = 5), Grade III: 5.2% (n = 6) vs 3.5% (n = 4), all P > 0.05]. The postoperative CGQL results showed a much better quality of life than preoperative one., Conclusion: As the first-choice operation for UC, RP-IPAA is safe and it offers satisfactory long-term outcomes and improved quality of life.

Objective: To investigate the imaging features, clinical manifestations and pathological characteristics of solitary fibrous tumors (SFT)., Methods: The clinicopathological manifestations and medical imaging findings were analyzed retrospectively in 27 patients with surgically confirmed SFT., Results: The SFTs originated from different parts of the body, including 18 in the chest, 4 in the abdomen, 1 in the lumboscral area, 3 in the pelvis, and 1 in the left shoulder. Twenty-three cases were found by CT scan, among which there were 16 benign diseases, presented with well-defined round or elliptic margins, with homogeneous attenuation and clearly surrounding; 6 malignant cases with unclear demarcations, invasive surrounding, heterogeneous attenuation due to calcification and/or irregular necrosis, and 1 junctional case with well-defined margins, which was enlarged during follow-up. There were 4 SFTs scanned by MRI with clear margin and homogeneous or heterogeneous signal intensity. All of the 4 cases were isointense or hyperintense to muscle on T1-weighted images, and were hyperintense on the T2-weighted images. All tumors showed heterogeneously intense enhancement with geographic pattern. Immunohistochemical staining showed that CD34-positive was 81.5%, vimentin (100.0%), CD99 (100.0%) and bcl-2 (96.3%), as well as negative CK (100.0%) and S-100 (96.3%)., Conclusion: The location of SFT is varying. Though its clinical manifestations vary, the diagnosis is depended on pathology and immunohistochemistry. There are certain specific features related to SFTs on CT or MRI. These imaging techniques may serve to provide helpful information as to the location and vicinal anatomic structure of the tumor, which is of substantial importance for planning surgery.

Aim: To construct a siRNA plasmid to knockdown Coronin-1., Methods: The cDNA of coronin-1 was amplified by RT-PCR from the total RNA of macrophage, and then inserted into pSEB-HUS vector to generate pSEB-HUS-C plasmid. Three synthesized siRNAs targeting Coronin-1 were cloned into pSEB-HUS-C respectively, resulting in the pSEB-HUS-C1, pSEB-HUS-C2 and pSEB-HUS-C3 plasmids. These plasmids were transiently transfected into A549, and the Coronin-1 level was detected by RT-PCR, Real time PCR and Western blot., Results: All plasmids were successfully constructed as confirmed by restriction enzyme digestion and DNA sequencing. The pSEB-HUS-C3 vector had the most significant knockdown effect on Coronin-1, with 75.9% inhibition at mRNA level and 75.1% inhibition at protein level., Conclusion: A siRNA plasmid targeting Coronin-1 was successfully constructed and validated for its knockdown effect, which will serve as a loss-of-function tool for the further mechanistic study of Coronin-1 in tuberculosis pathology.

Objective: The aim of this study was to evaluate the value of diffusion weighted imaging (DWI) in the diagnosis of patients with breast diseases., Methods: Fifty-three consecutive patients were scanned with GE signa HDx 1.5 T magnetic resonance system equipped with 8-channel breast coil. DWI was scanned by SE-EPI sequence in b values of 500 s/mm(2) and 800 s/mm(2), respectively. The apparent diffusion coefficients (ADC) of these lesions were measured. The mean apparent diffusion coefficients (ADC) of these lesions were calculated in b values of 500 s/mm(2) and 800 s/mm(2), respectively. These lesions' ADC value (rADC) was counted respectively and the result of the rADC was equal to the lesion's ADC divided by the ADC of the ipsilateral normal breast tissue. Threshold of ADC and rADC for differential diagnosis was acquired by ROC (receiver operating characteristic curve) analysis. Different imaging technologies were evaluated emphasizing their sensitivity, specificity and accuracy., Results: Sixty-six lesions of 53 cases were confirmed by pathology, including 39 malignant lesions and 27 benign lesions. (1) b = 500 s/mm(2), the threshold of ADC value was 1.435 x 10(-3) mm(2)/s, with a sensitivity of 82.1% and a specificity of 81.5%. The threshold of rADC value was 0.62, with a sensitivity of 76.9% and a specificity of 100%. (2) b = 800 s/mm(2), the threshold of ADC value was 1.295 x 10(-3) mm(2)/s, with a sensitivity of 79.5% and a specificity of 81.5%. The threshold of rADC value was 0.71, with a sensitivity of 89.7% and specificity of 88.9%. (3) The area under the ROC curve was increased for the four diagnostic indicators (ADC(500), ADC(800), rADC(500), rADC(800))., Conclusion: DWI spends short time, and it doesn't need contrast material. ADC value and rADC value have a high sensitivity and specificity as a diagnostic indicator. DWI is helpful in improving the specificity of MR and may become one of valuable conventional procedures for breast tumor diagnosis.

Concentrations of CO2, N2O and CH4 of soil profile in Karst Stone Desertification Areas were measured monthly from June of 2006 to May of 2007. The results indicated that the concentrations of CO2, N2O and CH4 varied from 530.2 to 31512.6, 0.27 to 0.67 and 0.1 to 3.5 microL x L(-1). On the whole, with the increase of soil depth, the concentrations of CO2 and N2O increased, while CH4 decreased. However, in the cold and wet October, November and January, the concentrations of CO2 and N2O decreased with the depth below 15 cm. In contrast, CHR4 had the inverse trend. Soil temperature and moisture were the main factors which had impacts on the temporal and spatial variations of C2O, N2O and CH4 concentrations in soil profile, but the influence impacts were different. The correlation analysis showed that there was positive correlation between CO2 and N2O (r 0.780-0.894, p < 0.05-0.01), while correlation degree was controlled by environmental factors in different months. CH4 concentrations in soil had significantly negative correlation with CO2 concentrations (r = 330, p < 0.01), but correlated with N2O concentrations just in moisture months (r - 0.829- - 0.956, p < 0.05-0.01).

The NSC-7 microbial community could decompose cellulose and lindan with high efficiency. In order to determine the bacterial composition of the community, 11 isolate strains were detected by plate isolation, while a community reset by the 11 isolate strains lost the capacity of degrading cellulose. The capacity of degrading of the filter paper in double deck plate and monolayer plate were determined, only the filter paper in double deck plate were degraded, that means the main or key microbe are anaerobic. The denaturing gradient gel electrophoresis (DGGE) and construction of 16S rDNA clone library were used to identify the composition diversity of NSC-7 community. 195 clones and 25 strains were detected in clone library, and about 60% closest relative among them was known the detailed information which were belonged to Clostridium, Petrobacter, Bacteria, Paenibacillus, Proteobacterium. Furthermore, there were 40% closest relative belonged to uncultured bacterium clone.

An investigation was carried out to approach the degradation of cyanide and maturity during the cassava processing wastes composting process. Mixtures of cassava hull, cassava residues and pig manure were used in the experiment. Parameters like temperature, pH, cyanide, cellulose, hemicellulose, lignin and C/N ratio were assessed during the composting process, the effect of composting process on the degradation of cyanide and maturity were evaluated. The results reveal that the content of cyanide decreases sharply and declines to 2.08 mg/kg (30 days of composting), the degradation rate of cyanide is 94.16% and is in accord with food safety standard. After 15 days of the composting process, degradation of composting materials containing carbon (starch, cellulose, hemicellulose) and cyanide are quick and the degradation rates of them are more than 80%, properties tend towards stability basically. During 30 days of the composting process, the composting temperature drops to normal temperature and tends to stability, pH remains stable at 7.2. Parameters like C/N ratio, nitrate-nitrogen (NO3(-)-N) and ammonia nitrogen (NH4(+)-N) as maturity evaluation index were measured, and the results indicate that physical and chemical properties keep stability after 15 days of cassava processing wastes composting process. At the end of fermentation, C/N ratio is 17.55, the content of nitrate-nitrogen and ammonia nitrogen reach 2.5g/kg and 10 mg/kg respectively, NO3(-)-N/NH4(+)-N ratio is 250. The changes of these above mentioned parameters meet with maturity evaluation standard. Proving that cassava processing wastes during 30 days of composting treatment can achieve stability and security state.

By using close chamber-GC method, the N2O fluxes in de-farming grassland, bush land, Pinus massoniana forestland, and broadleaved forestland in central Guizhou were measured in situ in 2006 and 2007 to investigate the seasonal change patterns of soil N2 O flux under different vegetations in karst region of southwest China. The results indicated that in a whole year round, the soils in all test sites played the source of atmospheric N2O, only acting as a N2O sink in exceptional months. The soil type and the vegetation had obvious effets on the N2O flux from soil. Yellow soil had a smaller N2O flux than calcareous soil, except for broadleaved forestland. The N2O fluxes from de-farming grassland and bush land were higher in spring and lower in autumn and winter, and varied within the range of -20.7 - 103.09 microg N x m(-2) x h(-1) and -33.0-67.3 microg N x m(-2) x h(-1), respectively; while those from P. massoniana forestland and broadleaved forestland were the highest in spring but had no definite change patterns in other seasons, and varied within the range of -5.3 - 35.0 microg N x m(-2) x h(-1) and -14.4 - 152.8 microg N x m(-2) x h(-1), respectively. The correlation analysis showed that there existed a negative correlation between soil moisture content and N2O flux, suggesting that soil moisture was the main factor driving the seasonal change of N2O flux. Temperature indirectly affected the soil N2O flux through affecting soil moisture.

To detect the metabolic characteristic of rice straw degradation by composite microbial system WSC-6, we cultured WSC-6 in the media used rice straw as the limiting carbon source. The rice straw was added in the style of different quantity once or the same quantity at the different time intervals during 90 days culture. The systems were cultivated under static condition at 50 degrees C. The degradation ratio, absolute degradation quantity,products from degradation and dynamics of pH value of fermentation system were all investigated. The results showed: when 1% rice straw was added once, the pH of fermentation system decreased from initial 7.8 to 6.0 within the first three days inoculation, and after six-day cultivation, it increased to 8.0 and was stable. For dissolved oxygen concentration (DO), the value was maintained at range of 0.01 to 0.12 mg x L(-1) of microaerobic condition. During the rice straw degradation, more than ten kinds of products including ethanol, acetic acid, lactic acid and glycerol and so on were detected using GC-MS. Especially, the highest concentration of lactic acid among all products was 7.381 g x L(-1) at 24 h after inoculation. During 90-day cultivation, for the addition treatments of the different quantity once, the more rice straw added, the quicker and lower the pH decreased, and the longer time intervals returned the pHs were. Especially for 5.0% addition, when 5.0% of rice straw was added once, pH did not increase again after it decreased. Among the addition of the same quantity at the different time intervals, the trend of decrease-increase in pH at 12-day and 15-day intervals could be repeated and high degradation activity well maintained. After 90-day of inoculation, the highest degradation ratio occurred in the treatment at 15-day interval, which was 86.7%. The highest absolute quantity occurred in the treatment at 6-day interval, which was 32.4 g. The trend of pH changes can indicate the activity of lignocellulose degradation and degradation process of the WSC-6.

The partial pressure of carbon dioxide p(CO2) in the surface water of the two karst eutrophic reservoirs, Hongfeng and Baihua Lakes near Guiyang city, Ghuizhou Province, China has been studied for one year. The factors regulating the seasonal variability of p(CO2) in the two reservoirs have been analyzed. Unlike the reservoirs in boreal temperate regions, the p (CO2) in the two reservoirs has significant seasonal changes: CO2 was unsaturated during summer, but supersaturated at other seasons. According to the correlation analysis, there is a negative relationship between p (CO2) and Chla in the two reservoirs, due to the production of the phytoplankton and the respiration of the organic matter by the bacteria, which are also the main factors regulating the p(CO2). The negative relationship between temperature and p(CO2) is considered to be due to the effect of temperature on the growth of the phytoplankton. Precipitation leads to the input of the nutriments into the reservoirs, and hence precipitation is negatively related to p(CO2). The relationships between NO3-, NO2- and p(CO2) are ascribed to the absorption of phytoplankton, and the degradation of the organic matter nitrification. The relationship between SiO3(2-) and p(CO2) is due to the absorption of phytoplankton and the precipitation. The differences of the relationship between p(CO2) and DOC in the studied two lakes possibly lie in the variations of their sources.

Aimed to utilize rice straw and lessen the pressure of environment, the rice straw was used as the fermentation material, and a lactic acid bacteria community SFC-2 from my laboratory was inoculated into the rice straw to investigate the inoculation effects. After 30 days fermentation, the inoculated fermented straw smelt acid-fragrant, and the pH value was 3.8, which was lower than the control of 4.1. Furthermore, lactic acid concentration was more than that in the control. Especially L-lactic acid concentration was two times more than in the control, and the crude protein content was 10.16% higher than that in the control, and the crude fiber content was 3.2% lower than that in the control. From the patterns of denaturing gradient gel electrophoresis (DGGE), Lactobacillus plantarum, Lactobacillus fermentum and Lactobacillus paracasei rapidly became the advantageous species in the inoculated straws. However, Enterobacter sakazakii, Pantoea agglomerans, Enterobacter endosymbiont, Pantoea ananatis, whichwere predominate in the controls, were not detected in the inoculated straws, and the fermented quality was improved significantly.

Objective: To synthesize a novel gadolinium-loaded nanoparticle as a molecular imaging contrast agent for magnetic resonance imaging., Methods: Gadolinium ion was incorporated within a silica nanoparticle. The size of this nanosized particle was determined by using transmission electron microscope. The spin-echo (SE) images of nine nanoparticle dilutions in vitro were obtained from a 1.5 T clinical scanner, compared with gadolinium diethylene triaminepenta acetate (Gd-DTPA). In vivo distribution of nanoparticle in Balb/c nude mice and Balb/c nude mice with nasopharyngeal carcinoma (NPC) xenografted CNE-2 tumors was studied at MR imaging, 30 sec, 5 min, 30 min, 1 h, 2 h, 4 h, and 24 h after intravenous administration., Results: The gadolinium-loaded nanoparticle was short rod-shaped, and approximately 30 to 40 nm in diameter. The value of longitudinal relaxativity (r(1)) of gadolinium nanoparticle was much higher than that of Gd-DTPA. Thirty minutes after injection the gadolinium nanoparticle, the signal intensities of liver, kidney and xenografted tumor increased from 226 +/- 10, 283 +/- 7 and 195 +/- 5 to 352 +/- 12, 328 +/- 10 and 245 +/- 7, respectively. The dynamic MRI scanning showed that gadolinium nanoparticles were mainly distributed in liver after intravenous administration. Strong enhancement was also detected in CNE-2 xenografted tumors., Conclusion: A new gadolinium-loaded nanoparticle with high relaxativity was synthesized successfully, and might serve as a carrier for magnetic resonance molecular imaging.

In this study, histone H1, core histones H2A-H2B and H3-H4 were purified from chicken erythrocytes by hydroxylapatile chromatography. The nuclear extract was prepared from HeLa cells. We investigated the binding and interaction of histones and transcription factors on the upstream sequence of human autocrine motility factor receptor (hAMFR) gene by gel shift mobility assay. We found that the binding of H1 on the promoter sequence of hAMFR gene was relatively stable. We propose that H1 plays an important role in stablizing chromatosome. We also found that histones and HeLa cell extract could form a ternary complex with the DNA template.

The effects of interaction between histones and human autocrine motility factor receptor (hAMFR) gene promoter on the transcription activity in vitro was investigated by using histones purified from chicken erythrocytes, HeLa cell nuclear extracts and heat-treated supernatants of Xenopus eggs. The results showed that the competitive binding of histones and transcription factors at the promoter of hAMFR gene was very important to the transcription in vitro. If a pre-initiation complex was formed with HeLa cell nuclear extracts on the promoter prior to nucleosome assembly, it would prevent nucleosome-mediated transcription repression. When the nucleosome was assembled on the promoter in advance, the transcription activity could be repressed. When histones and HeLa cell nuclear extracts were mixed in the reaction simultaneously, the transcription activity would depend on the relative amount of histones to that of HeLa cell nuclear extracts.

We report on the culturing of melanocytes from suction blisters from the uninvolved skin of localized (focal and segmental) vitiligo patients and from the foreskins of newborns and adults, over a long period of time using a modified culture medium composed of F-12 medium supplemented with insulin (5 mg/mL), cholera toxin (40 ng/mL), transferrin (5 mg/mL), hydrocortisone (1 mM), epithelial growth factor (20 ng/mL), endothelial cell growth supplement (ECGS) (15 mg/mL), retinol (1 x 10(-7) M), 12-o-tetradecanoyl-phorbol-13 acetates (TPA) (85 nM), and 1% fetal calf serum (FCS). This method may be used in in vitro studies on normal human melanocytes and for study of the difference between melanocytes of normal individuals and of those with vitiligo. The ability to culture melanocytes from localized vitiligo, and the inability to grow those of generalized vitiligo with this system reconfirms the difference in the pathogenesis of various types of vitiligo. Thus, culturing may be used for differentiating localized vitiligo from generalized vitiligo and may be used to guide the mode of treatment when vitiligo has just started to develop and only a few depigmented patches have appeared. The results of studies on seeding density effect and serial Dopa reaction on the melanocytes of normal individuals reveal that the use of tyrosinase activity as the assessment parameter requires the use of melanocytes cultured for less than five months at a seeding density of less than 4 x 10(4) cells/cm2. Enlarged and heavily pigmented cells, senescent cells, were observed after very long-term culture (one year and four months).(ABSTRACT TRUNCATED AT 250 WORDS)

To understand the clinical characteristics of trichotillomania, 36 patients were investigated at the psychocutaneous special clinic from February 1982 to October 1990. There were 16 males and 20 females. The most prominent age was found in a group of 23 (63.9%) elementary school children, and in regards to the age of disease onset, 35 (97.2%) of our patients were under the age of 18. A 30-year-old woman with schizophrenia and a 6-year-old boy with mental retardation were noted in our study; the remaining 34 patients were generally well without signs of mental illness except for the recurrent failure to resist impulses to pull out their own hair. The common life events precipitating stress were academic problems and parent-child conflicts. The duration of follow-up was for at least 6 months. The habit of hair plucking was easily corrected if the duration of the disease was less than 6 months. Usually these patients had a satisfactory regrowth of hair within 1-3 months after our explanation and reassurance. In contrast, the patients with a hair loss of more than half a year were not easy to approach, and needed psychiatric treatment.

Cryofibrinogenemia, a rare disease, is diagnosed by the presence of reversible cryoprecipitate in plasma, which may cause various clinical manifestations of thromboembolic or hemorrhagic disorders. A 66-year-old female patient suffered from recurrent reticulate erythema, purpura and gangrene on the distal portion of the extremities and face during the winter for 5 years. Cryofibrinogenemia was proven with the following evidences: (1) Histopathologic findings revealed many eosinophilic cylinders, about 6-9 microns in diameter, deposited within the lumen of the blood vessels in the dermis and subcutaneous tissue. There was no evidence of vasculitis. These cylinders were fibrinogen in nature, as proven by direct immunofluorescence. (2) Cryofibrinogen was detected in the citrated plasma, which was confirmed by double immunodiffusion, the presence of an IgG-kappa M protein was also revealed by immunofixation. Debridement and skin grafts were performed, and prednisolone combined with cyclophosphamide were given. Skin lesions improved without recurrence, and the "cryocrit" decreased from 13.6% to less than 2% during the follow up. We conclude that if there is any case with the similar cutaneous manifestations, cryofibrinogenemia should be considered.

Ciclopiroxolamine is now under use as an antifungal agent for about a decade. Its activity is well-known in moderate climates, but there is a lack of knowledge on the activity in tropical and subtropical areas. 49 patients suffering from onychomycosis (42) and/or tinea pedis (33) were enrolled in a clinical trial in Taiwan. The patients received ciclopiroxolamine 1% cream 2-3 times a day for 3 to 24 month. In addition, infected nails were filed to facilitate drug penetration. In onychomycosis, 14% of the patients were cured and another 36% improved. One patient relapsed after 10 months. Cure was achieved more easily, if there were only modest alterations of the nail plate. With respect to tinea pedis the results were more favourable, the cure rate was 42%, another 45% of the patients improved. Following this dosage regimen, side effects did not occur. The results show onychomycosis to be highly resistant to antifungal therapy, perhaps even more as compared to tinea pedis in temperate climates. The good tolerability favours topical ciclopiroxolamine for this recalcitrant disease.

To clarify the mechanism of immunotoxicity of polychlorinated biphenyls (PCB), delayed type hypersensitivity skin test, lymphoproliferative responses, and lymphocyte subpopulations defined by monoclonal antibodies were performed on 30 patients and 38 age/sex-matched healthy individuals. Three years ago the patients had ingested rice bran oil accidentally contaminated by PCB. Except for lymphocyte proliferation, the chronic effect of PCB and its derivatives upon cell-mediated immunity (CMI) in these patients was not so evident when compared to that found within one year of exposure. Delayed type hypersensitivity skin response to purified protein derivative (PPD) and the numbers of peripheral T and B lymphocytes were within normal range. The enumeration of lymphocyte subsets demonstrated decreased T4 (inducer/helper) cells and increased T8 (cytotoxic/suppressor) cells. The abnormally augmentated spontaneous proliferation and proliferation responses to phytohemagglutinin (PHA), concanavalin A (Con A), pokeweed mitogen (PWM) and PPD implicated that both T and B lymphocytes were functionally affected. This may well be the result of the immunotoxicity rather than the enhancement of cell-mediated immunity by these toxic compounds. While there was no correlation between the results of those assays and blood PCB levels, it appears that the lymphoproliferative assay may be applied as a sensitive test in reflecting the severity of PCB immunotoxicity.

The current hypothesis on the pathogenesis of alopecia areata is immunologically mediated, yet no conclusive data has been documented. To clarify the changes in various phases of disease evolution, analysis of T lymphocyte subpopulations was carried out in the peripheral blood of 34 patients and 27 healthy normal subjects by means of OKT-3, OKT-4, OKT-8 and Ia1 monoclonal antibodies. Study of lymphocyte proliferation in response to various mitogens was further carried out in 27 patients and 25 healthy normal subjects. Significant predominance (p less than 0.01) of OKT-8 positive (cytotoxic/suppressor) cells and diminution (p less than 0.01) of Ia1 positive cells were noted in the whole alopecia group and in various subgroups of disease evolution. Since diminution (p less than 0.05) of OKT-4 positive (inducer/helper) cells was also noted in the stable phase, it was postulated that these activated T cells migrated from circulation to the local site of primary event. Hence, it was evident that the predominance of cytotoxic/suppressor cells was a relative change, and the role of inducer/helper cells in the pathogenesis was implicated. No enhancement of lymphocyte proliferation was observed.

To evaluate the clinical applicability of anti-native DNA antibodies (anti-nDNA) in systemic lupus erythematosus (SLE), two conventional methods--the Farr radioimmunoassay (Farr assay) and the Crithidia luciliae immunofluorescent technique (CL-IF assay)--were compared in 180 sera of antinuclear antibody (ANA)-positive systemic lupus erythematosus (85 active and 95 inactive), 31 sera of ANA-positive non-SLE rheumatic diseases and 40 sera from healthy subjects. The results further appraised the clinical significance of anti-nDNA antibodies in the diagnosis of systemic lupus erythematosus, especially those patients with nephritis. The CL-IF assay was found less sensitive in the detection of anti-nDNA than the Farr assay. As a tool for differentiating SLE from non-SLE rheumatic diseases, the CL-IF assay is not as sensitive as the Farr assay (sensitivity of CL-IF and Farr assay = 38.3% versus 60.0%). But the CL-IF test is much more convenient and efficient in the detection of active stage SLE and lupus nephritis than the Farr assay. The efficiency of anti-nDNA tests by the CL-IF and Farr assays for diagnosing active SLE were 85.8% versus 79.6%, for lupus nephritis were 76.1% versus 64.4% and finally for both conditions were 81.7% versus 62.2%. As an index of disease activity during the course of treatment, titers of anti-nDNA antibodies by Farr assay proved much more reliable than those by Crithidia luciliae assay. For the detection of anti-nDNA antibodies, simultaneous utilization of these two methods for fresh cases and using the Farr assay alone during the course of treatment are recommended.

A family in Taiwan is reported to have Mal de Meleda with possibly autosomal recessive inheritance among 5 of its members. The glove- and sock-like erythrokeratosis and the hyperkeratotic plaques of the knees and elbows are the common features. Erythema of the nose, cheeks, and the perioral area in 4 affected members, actinic elastosis in 2 severely affected members, and partially impaired cell-mediated immunity in vitro in both affected and unaffected members are notable findings. Tigason (etretinate) treatment was beneficial in regard to hyperkeratinization, but not to erythema.

An outbreak of poisoning caused by ingestion of rice bran oil which was accidentally contaminated with polychlorinated biphenyls (PCBs) broke out in Taiwan in February 1979. Diagnosis, management, and follow-up of the patients were performed at special clinics, and subjective symptoms and cutaneous changes such as peculiar acneform eruptions and pigmentation were recorded. The patients were divided into six age groups of both essex, and the body surface of the patients was divided into 12 sections according to the nature of skin. The prevalence of each type of cutaneous change was proved statistically by the chi-square test. The examination of the immune system function in the patients at 1 year revealed: decreased concentration of IgM and IgA but not of IgG; decreased percentage of total T-cells, active T-cells, and helper T-cells, normal percentage of B-cells and suppressor T-cells; suppression of delayed type response to recalling antigens; enhancement of lymphocyte spontaneous proliferation; and enhancement of lymphocyte proliferation with PHA, PWM, and PPD stimulation but not ConA. Follow-up studies 3 years later showed decreased blood PCB levels; some improvement of subjective symptoms and cutaneous changes; recovery of skin testing response to PPD; normal percentage of total T-cells and increased percentage of suppressor T-cells; and enhancement of lymphocyte proliferation spontaneously or under the stimulation of various mitogens.

Prevalence of cytoplasmic antibodies--smooth muscle antibodies (SMA), gastric parietal cell antibodies (GPA), and mitochondrial antibodies (MTA)--was evaluated in 148 normal persons and 168 patients by indirect immunofluorescent method. Their prevalence in normal persons was 0%, 2% and 0% for SMA, GPA and MTA respectively, while SMA and MTA were positive in 5.7% and 8.6% of the 35 systemic lupus erythematosus (SLE) patients respectively. The difference in the prevalence of SMA and MTA between these two groups was statistically significant. The higher prevalence of these antibodies and the occurrence of various kinds of antibodies in SLE patients support the thesis that SLE is an autoimmune phenomenon.

One hundred and ten cases of an unusual type of naevus, which we have called naevus fusco-caeruleus zygomaticus were studied. The naevus presents as a bilateral speckled discolouration of the skin of the face principally in the zygomatic region. The condition usually does not become apparent until the second decade of life, and is much commoner in females. No association with any other abnormalities was found. Light and electron microscopy of the speckles showed the presence of dermal melanocytes mostly in the upper dermis. A survey was also carried out to determine the prevalence of this naevus in the general population.

This paper describes in detail the dermatological, medical, and laboratory findings of patients poisoned with PCBs and related compounds in Taiwan. Together with their symptoms, their skin lesions as well as their hematological, immunological, and other clinical features are explained. A grading of their clinical severity was tried, and its possible association with PCB concentrations in their blood was examined but could not be demonstrated. Various treatments have been applied but without notable success. Even so, a follow-up study of patients one year later showed that about 38% of patients were somewhat clinically improved, while 54% were not altered.