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The in situ and real-time supervision of reactive oxygen species (ROS) generated during photodynamic therapy (PDT) is of great significance for lessening nonspecific damage and guiding personalized therapy. However, photosensitizers frequently fail to deliver successful treatment accompanying the ROS-related imaging signals produced, impeding simple treatment outcome predictions and therapeutic schedule adjustments. Here, we report a two-photon fluorescence self-reporting strategy for the in situ and real-time monitoring of treatment response via a novel black phosphorus-based two-photon nanoprobe (TPBP). TPBP effectively generated singlet oxygen (1O2) under near-infrared laser irradiation for PDT, and 1O2 stimulated a two-photon molecule to emit fluorescence signals for feedback of 1O2 generation, which facilitated the regulation of treatment parameters to achieve precise and personalized medicine in deep tissue.

Furin, an important member in the family of proprotein convertases, is a participant in the activation of various precursor proteins. The expression level of furin stays in a very low range in most normal cells, but elevates with a big margin in many cancer cells. More importantly, furin is closely related to tumor formation and migration. Herein, a furin-activatable near-infrared (NIR) fluorescent probe (HD-F) was first developed that allowed for specific, sensitive detection and imaging of furin both in vitro and in vivo. HD-F consists of a classical NIR fluorophore (HD), a furin-particular polypeptide sequence RVRR, and a self-eliminating linker. Without the interaction with furin, no noticeable fluorescence enhancement was detected, even over 3 days, demonstrating the excellent stability of HD-F. Upon conversion by furin, there was a distinct signal increase around 708 nm. It has achieved assay and visualization of endogenous furin in various cells, tumor tissues, and tumor-bearing mouse models. Importantly, HD-F is well-suited for monitoring the change of furin expression level in the process of hypoxia-inducible factor-1 stabilized by CoCl2. Moreover, HD-F could visualize the divergence in the expression level of furin between normal and cancer cells, indicating its potential in specific cancer imaging. Thus, this novel probe is able to serve as a potential tackle for better understanding of the intrinsic link between a hypoxic physiological environment and cellular carcinogenesis and predicting cancer in preclinical applications.

Cell membrane-targeted bioimaging is a prerequisite for studying the roles of membrane-associated biomolecules in various physiological and pathological processes. However, long-term in situ bioimaging on the cell membrane with conventional fluorescent probes leads to diffusion into cells from the membrane surface. Therefore, we herein proposed a de novo strategy to construct an antidiffusion probe by integrating a fluorochrome characterized by strong hydrophobicity and low lipophilicity, with an enzyme substrate to meet this challenge. This precipitating fluorochrome HYPQ was designed by conjugating the traditionally strong hydrophobic solid-state fluorochrome 6-chloro-2-(2-hydroxyphenyl) quinazolin-4(3H)-one (HPQ) with a 2-(2-methyl-4H-chromen-4-ylidene) malononitrile group to obtain closer stacking to lower lipophilicity and elongate emission to the far-red to near-infrared wavelength. As proof-of-concept, the membrane-associated enzyme γ-glutamyltranspeptidase (GGT) was selected as a model enzyme to design the antidiffusion probe HYPQG. Then, benefiting from the precipitating and stable signal properties of HYPQ, in situ imaging of GGT on the membrane was successfully realized. Moreover, after HYPQG was activated by GGT, the fluorescence signal on the cell membrane remained unchanged, with incubation time even extending to 6 h, which is significant for in situ monitoring of enzymatic activity. In vivo testing subsequently showed that the tumor region could be accurately defined by this probe after long-term in situ imaging of tumor-bearing mice. The excellent performance of HYPQ indicates that it may be an ideal alternative for constructing universal antidiffusion fluorescent probes, potentially providing an efficient tool for accurate imaging-guided surgery in the future.

A FRET-based probe for mapping the fluctuation of ONOO− in cisplatin-induced acute kidney injury was constructed. It exhibits ratiometric near infrared fluorescence and a dramatic decrease of its peak absorbance at 719 nm upon addition of ONOO− that is converted into remarkable signal changes in fluorescence and photoacoustic images respectively.

Molecular probes activated by a single enzyme have been extensively used in bioimaging and disease diagnosis; however, imaging and identification in an accurate manner remains a challenge for such probes. Here, based on the specificity of enzyme recognition, we engineered a "double-locked" and enzyme-activated molecular probe (NML) for accurate bioimaging and hepatopathy differentiation. Triggered by the successive reactions with leucine aminopeptidase (LAP, first "key") and monoamine oxidase (MAO, second "key"), the emissive fluorophore (NF) was released. NML can be activated only in the presence of both LAP and MAO and can be silenced when either enzyme is inhibited. Benefiting from the "double-locked" strategy, NML showed higher accuracy for imaging of drug-induced liver injury (DILI) than the "single-locked" probe. With serum testing, NML showed significant differences in mouse models of both CCl4-induced liver cirrhosis and DILI. Significantly, NML can be applied to accurately distinguish serum samples from clinical patients with different hepatopathies. Our smart molecular probe may hold great potential for hepatopathy diagnosis and clinical transformation.

The ability to detect cancer early in an accurate and rapid fashion is of critical importance for cancer diagnosis and accurate resection in surgery. γ-Glutamyltranspeptidase (GGT) is overexpressed in several human cancers, while maintaining a low expression in normal microenvironments, and thus is recognized as an important cancer biomarker. To date, rational design of a zero cross-talk ratiometric near-infrared (NIR) GGT fluorescent probe for efficient cancer diagnosis in various biological samples is still a big challenge. In this work, a zero cross-talk ratiometric NIR GGT fluorescent probe named Cy-GSH is developed. Cy-GSH shows high sensitivity to GGT, which is desired for early cancer diagnosis. Upon additional GGT, a large emission shift from 805 to 640 nm is observed, which is suitable for visualizing deeply located cancer in vivo. In addition, successful monitoring of GGT activity in blood, cells, tissues, and in vivo makes Cy-GSH possess great potential for the clinical cancer early diagnosis. Furthermore, accurately visualizing tumors and metastases in mouse models illuminates that the probe may be a convenient tool for fluorescence-guided cancer surgery. To our knowledge, this is the first report to describe the strategy of a zero cross-talk ratiometric NIR GGT fluorescent probe for early cancer diagnosis and fluorescence-guided surgery.

Hydrogen sulfide (H2S) is involved in various pathological and physiological processes as well as regulation of lysosomal functions. Thus, it is essential to develop precise method for the detection of H2S in lysosome. Although a large number of fluorescent probes for H2S have been developed, the probe for lysosomal H2S assay still remains rare. Herein, a two-photon fluorescent probe, named Lyso-Nap, is designed and synthesized for detecting H2S in lysosome by employing a dual lock system (dual PET process and dual reaction sites). The fluorescence of Lyso-Nap with dual lock system is opened by H2S only when the probe is localized within acidic lososomes, which can avoid the disturbance from neutral cytosol and other organelles. Moreover, two-photon fluorescent probes with weak auto-fluorescence, large penetration depth and reduced photodamage, are particularly favored for bioimaging. Accordingly, Lyso-Nap can be successfully used for precisely tracking and detecting endogenous H2S in lysosome of living cells and tissue.

Peroxynitrite (ONOO–), an extremely reactive nitrogen species (RNS), is implicated in diverse pathophysiological conditions, including cancer, neurodegenerative diseases, and inflammation. Sensing and imaging of ONOO– in living systems remains challenging due to the high autofluorescence and the limited light penetration depth. In this work, we developed a bioluminescent probe BP-PN, based on luciferase–luciferin pairs and the ONOO–-responded group α-ketoamide, for highly sensitive detection and imaging of endogenous ONOO– in living cells and mice for the first time. Attributed to the BL without external excitation, the probe BP-PN exhibits a high signal-to-noise ratio with relatively low autofluorescence. Furthermore, we examine the application of the probe BP-PN using the mice model of inflammation, and BP-PN shows high sensitivity for imaging endogenous ONOO– in inflamed mice. This newly developed bioluminescent probe would be a potentially useful tool for in vivo imaging of ONOO– in wider physiologica...

Abnormal enzymatic activities are directly related to the development of cancers. Identifying the location and expression levels of these enzymes in live cancer cells have considerable importance in early-stage cancer diagnoses and monitoring the efficacy of therapies. Small-molecule fluorescent probes have become a powerful tool for the detection and imaging of enzymatic activities in biological systems by virtue of their higher sensitivity, nondestructive fast analysis, and real-time detection abilities. Moreover, due to their structural tailorability, numerous small-molecule enzymatic fluorescent probes have been developed to meet various demands involving real-time tracking and visualizing different enzymes in live cancer cells or in vivo. In this review, we provide an overview of recent advances in small-molecule enzymatic fluorescent probes mainly during the past decade, including the design strategies and applications for various enzymes in live cancer cells. We also highlight the challenges and opportunities in this rapidly developing field of small-molecule fluorescent probes for interventional surgical imaging, as well as cancer diagnosis and therapy.

Mitophagy induced by hypoxia plays an important role in regulating cellular homeostasis via the removal of dysfunctional mitochondria in the lysosomal degradation pathway, which results in physiological changes in the mitochondria, such as the pH, polarity and viscosity. However, the lack of an effective method for imaging of both the hypoxic microenvironment and the resulting variable mitochondria limits the visualization of hypoxia-induced mitophagy. Based on the specific mitochondrial pH changes during the hypoxia-induced mitophagy process, we have reported a near-infrared fluorescent probe (NIR-HMA) for real-time simultaneous visualization of the hypoxic microenvironment and the subsequent mitophagy process in live cells. NIR-HMA selectively accumulated in the hypoxic mitochondria in the NIR-MAO form, emitting at 710 nm, and then transformed into NIR-MAOH, emitting at 675 nm, in the acidified mitochondria-containing autolysosomes. Importantly, by smartly tethering the hypoxia-responsive group to the hydroxyl group of the NIR-fluorochrome, which shows ratiometric pH changes, NIR-HMA can differentiate between different levels of the hypoxic microenvironment and mitophagy. Furthermore, using NIR-HMA, we could track the complete mitophagy process from the mitochondria to the autolysosomes and visualize mitophagy caused only by hypoxia both in cancer cells and normal cells. Finally, NIR-HMA was applied to investigate the role that mitophagy plays in the hypoxic microenvironment via the cycling hypoxia-reoxygenation model. We observed a decreased fluorescence ratio after reoxygenation and a further increased mitophagy level after hypoxia was induced again, suggesting that mitophagy might be a self-protective process that allows cells to adapt to hypoxia. Our work may provide an attractive way for real-time visualization of relevant physiological processes in hypoxic microenvironments.

Oxidative stress, a disturbance in the balance between oxidant/antioxidant ratios, is associated with cancer, aging, inflammation, neurodegenerative diseases and other conditions. γ-Glutamyltranspeptidase (GGT) is a redox-related enzyme that plays a key role in mitigating the effects of oxidative stress by maintaining cellular glutathione (GSH) metabolism and homeostasis. Therefore, oxidative stress will upregulate the intracellular GGT level. To better understand the major pathophysiological resist mechanism to oxidative injury in mediating many disease states, we designed and synthesized a novel two-photon (TP) fluorescent turn-on probe, Np-Glu, for GGT detection and bioimaging. Under the optimized conditions, Np-Glu exhibited remarkable fluorescence enhancement (150-fold), good selectivity and high sensitivity (LOD is 0.033 U L-1), with a wide linear concentration range (0-50 U L-1). More importantly, the probe Np-Glu was successfully applied in one-photon and TP fluorescence imaging of GGT activity in an oxidative stress model in living cells and tissues, suggesting Np-Glu as an ideal indicator for clinical and biological samples.

Nontoxic prodrugs, especially activated by tumor microenvironment, are urgently required for reducing the side effects of cancer therapy. And combination of chemo-photodynamic therapy prodrugs show effectively synergetic therapeutic efficiency, however, this goal has not been achieved in a single molecule. In this work, we developed a mitochondrial-targeted prodrug PNPS for near infrared (NIR) fluorescence imaging guided and synergetic chemo-photodynamic precise cancer therapy for the first time. PNPS contains a NIR photosensitizer (NPS) and an anticancer drug 5′-deoxy-5-fluorouridine (5′-DFUR). These two parts are linked and caged through a bisboronate group, displaying no fluorescence and very low cytotoxicity. In the presence of H2O2, the bisboronate group is broken, resulting in activation of NPS for NIR photodynamic therapy and activation of 5′-DFUR for chemotherapy. The activated NPS can also provide a NIR fluorescence signal for monitoring the release of activated drug. Taking advantage of the high H2O2 concentration in cancer cells, PNPS exhibits higher cytotoxicity to cancer cells than normal cells, resulting in lower side effects. In addition, based on its mitochondrial-targeted ability, PNPS exhibits enhanced chemotherapy efficiency compare to free 5′-DFUR. It also demonstrated a remarkably improved and synergistic chemo-photodynamic therapeutic effect for cancer cells. Moreover, PNPS exhibits excellent tumor microenvironment-activated performance when intravenously injected into tumor-bearing nude mice, as demonstrated by in vivo fluorescence imaging. Thus, PNPS is a promising prodrug for cancer therapy based on its tumor microenvironment-activated drug release, synergistic therapeutic effect and “turn-on” NIR imaging guide.

Acute organ injury observed during sepsis, caused by an uncontrolled release of inflammatory mediators, such as lipopolysaccharide (LPS), is quite fatal. The development of efficient methods for early diagnosis of sepsis and LPS-induced acute organ injury in living systems is of great biomedical importance. In living systems, cystathionine γ-lyase (CSE) can be overexpressed due to LPS, and H2Sn can be formed by CSE-mediated cysteine metabolism. Thus, acute organ injury during sepsis may be correlated with H2Sn levels, making accurate detection of H2Sn in living systems of great physiological and pathological significance. In this work, our previously reported fluorescent platform was employed to design and synthesize a FRET-based ratiometric two-photon (TP) fluorescent probe TPR-S, producing a large emission shift in the presence of H2Sn. In this work, a naphthalene derivative two-photon fluorophore was chosen as the energy donor; a rhodol derivative fluorophore served as the acceptor. The 2-fluoro-5-nitr...

Placental trophoblast differentiation involves the continuous fusion of mononuclear cytotrophoblasts. However, except for syncytin, little is known about the detailed mechanisms underlying trophoblast fusion. A previous study indicated that lipid rafts play an important role in HTLV-1 syncytium formation. To identify proteins that may be involved in placental trophoblast differentiation, we examined stomatin, an important lipid-raft protein that localizes to detergent-resistant membrane domains. The syncytium and human chorionic gonadotropin (β-hCG; a marker of placental trophoblast differentiation) were visualized by immunofluorescence staining. We found that overexpression of stomatin in the nonfusogenic JEG-3 cell line caused syncytium formation and increased the fusion index of cells. Treating these cells with N(6) ,2'-O-dibutyryladenosine 3',5'-cyclic monophosphate further increased cell fusion by stomatin. β-hCG was found in a few JEG-3 cells overexpressing stomatin at 48 h, and its levels increased dramatically at 72 h along with the formation of the multinuclear syncytium. RNA interference was used to decrease stomatin expression in BeWo cells, a fusogenic human choriocarcinoma cell line. After knockdown for 72 h, stomatin levels decreased by almost 95%. The fusion indexes of control and stomatin-knockdown cells at 72 h were 9.4 and 6.5%, respectively. Our data indicated that stomatin could trigger syncytium formation and upregulate β-hCG for cell fusion in nonfusogenic JEG-3 cells. Downregulation of stomatin slightly inhibited the fusion index of fusogenic BeWo cells. Thus, these data suggested that stomatin plays an important role in trophoblast differentiation.

A promising two-photon fluorescent probe MNAH for detecting 1O2 during the PDT process in mitochondria was proposed for the first time. MNAH was successfully applied for two-photon imaging of 1O2 in living cells and tissues during the PDT process with deep-tissue imaging depth. MNAH can be a powerful molecular tool for studying 1O2 generation in mitochondria during the PDT process.

Cellular self-regulation of reactive oxygen species (ROS) stress via antioxidant repair plays an important role in maintaining the redox balance. The redox balance between reducing and oxidizing species within cells is significant in the regulation of a signal pathway and is achieved by a series of elaborate mechanisms. In this work, we employed our previously reported D–π-A-structured naphthalene–BODIPY TBET platform to design an efficient two-photon fluorescent probe for dynamic monitoring of superoxide anion oxidative stress and the GSH reducing repair process. The probe displayed high energy transfer efficiency (91.4%), large pseudo-Stokes shifts upon one-photon excitation, and red fluorescence emission (λem = 596 nm), which is highly desirable for bioimaging applications. The probe exhibits reversibility, rapid response, good photostability, high selectivity and sensitivity for the superoxide anion and GSH. More importantly, the probe was successfully applied for visualizing the redox changes in living cells and tissues.

Drug-induced liver injury (DILI) is a potential complication of any prescribed medication. So far, the diagnosis of DILI is still a clinical challenge due to the lack of efficient diagnosis method. Glutathione S-transferase (GST), with a high concentration in liver cytosol, can reduce toxicity and facilitate urinary excretion by catalyzing the conjugation of glutathione (GSH) with reactive metabolites in liver. When liver is seriously damaged, GST and GSH will be released into plasma from liver cytosol, which caused a lower GST activity in liver cytosol. Therefore, monitoring the level of GST activity in liver tissue may be a potential strategy for diagnosis of DILI. Here, we reported a two-photon probe P-GST for GST activity detection for the first time. In the proposed design, a donor-π-acceptor (D-π-A) structured naphthalimide derivative with efficient two-photon properties was chosen as the fluorescent group, and a 2,4-dinitrobenzenesulfonate group was employed as the GST recognition unit, which also acted as the fluorescence quencher. In the present of GST and GSH, the recognition unit was removed and the fluorophore was released, causing a 40-fold enhancement of fluorescence signal with a detection limit of 35 ng/mL. At last, P-GST was successfully applied in two-photon imaging of GST in cells and DILI samples, which demonstrated its practical application in complex biosystems as a potential method for diagnosis of DILI.

Systemic lupus erythematosus (SLE) is a systemic autoimmune disease primarily affecting women. Previous studies have indicated that sex hormone estrogens contribute to the female predilection of SLE. Estrogen regulates gene expression by translocating estrogen receptors (ER) α and β into the nucleus where they induce transcription by binding to estrogen response elements of target genes. We have previously observed that expression of ERα gene and protein in lupus patients is significantly higher than in healthy controls and that estradiol up-regulates calcineurin expression via over-expression of ERα gene in SLE. However, the pathogenesis of over-expression of ERα gene is unknown. Here we report that enhanced expression of ERα mRNA and protein in SLE and rheumatoid arthritis is associated with DNA demethylation within the proximal promoter region located between -232 and +81 base pair relative to transcription start site of human ERα gene (GenBank Accession no. AL356311.6). The frequency of DNA demethylation was comparable between male and female. These findings suggest that estrogen and demethylated ERα promoter associated up-regulated ERα genes are two critical factors in the gender biased development of autoimmune diseases besides genetic factor.

Focal contacts, large macromolecular complexes that link the extracellular matrix and the internal cell cytoskeleton, are thought to govern cell locomotion. However, the maturation process through which focal contacts control the cellular migratory machinery by changes in size and molecular composition remain unclear. Here, we fabricated cell growth substrates that contained linear ECM strips of micron- or submicron-width in order to limit the enlargement of focal contacts. We found that NBT-II cells plated on the submicron substrate possessed smaller focal complexes that exhibited a highly dynamic turnover. These cells possessed various leading edges at multiple sites of the cell periphery, which prevented the cell from advancing. In contrast, cells grown on the micron-width substrate possessed large and stable focal adhesions. Most of these cells were elongated bipolar cells that were tethered at both ends and were immobile. Further, EGF and ROCK signaling pathways can modulate the cellular migratory responses according to the substrate guidance. On the submicron-width substrate, EGF treatment increased the focal contact size and the contractile force, causing these cells to develop one leading edge and migrate along the submicron-sized ECM paths. In contrast, inhibition of ROCK signaling decreased the focal contact size for cells plated on the micron substrate. These cells became less tethered and were able to migrate along or even across the micron-sized ECM paths. Our results indicate that formation and maturation of focal contacts is controlled by both ECM cues and intracellular signaling and they play a central role in directed cell motion.

Systemic lupus erythematosus (SLE) is an autoimmune disease primarily affecting women (9:1 compared with men). To investigate the influence of female sex hormone estrogen on the development of female-biased lupus, we compared the expression of estrogen receptor alpha (ERα) gene and protein levels as well as expression of T-cell activation gene calcineurin in response to estrogen in peripheral blood lymphocytes (PBLs) from SLE patients and normal controls. PBLs were isolated from 20 female SLE patients and 6 normal female controls. The amount of ERα protein in PBL was measured by flow cytometry. The expression of ERα and calcineurin messenger RNA was measured by semi-quantitative reverse transcription-polymerase chain reaction. Calcineurin phosphatase activity was measured by calcineurin assay kit. The expression of ERα messenger RNA and ERα protein was significantly increased (p = 0.001 and p = 0.023, respectively) in PBL from SLE patients compared with that from normal controls. In addition, the basal calcineurin in PBL from SLE patients was significantly higher (p = 0.000) than that from normal controls, and estrogen-induced expression of calcineurin was increased (p = 0.007) in PBL from SLE patients compared with that from normal controls, a 3.15-fold increase. This increase was inhibited by the ERα antagonism ICI 182,780. The effects of ER antagonism were also found in calcineurin activity. These data suggest that overexpression of ERα gene and enhanced activation of calcineurin in response to estrogen in PBL may contribute to the pathogenesis of female dominant in SLE.

To investigate the associations of DNA methylation levels and mRNA expressions of DNA cytosine-5-methyltransferase 1 (DNMT1) and methyl CpG-binding domain 2 (MBD2) with rheumatoid arthritis (RA).The global methylation status of DNA was measured in 65 patients with RA and 64 healthy controls by the ELISA method. DNMT1 and MBD2 mRNA were also detected in 177 RA patients and 95 controls using the quantitative real-time polymerase chain reaction method.The global methylation of DNA was significantly decreased in the RA patients compared to the controls (p=0.005, 95% CI=0.0835-0.4503). The patients with RA had higher expressions of DNMT1 and MBD2 mRNA than the controls (p0.001, 95% CI=-0.0024 to -0.0053 and p0.001, 95% CI=-0.0079 to -0.0167, respectively). We also found that the MBD2 mRNA level was not related to the disease activity of RA. However, the expression of DNMT1 mRNA tended to be associated with the disease activity of RA (p=0.08). The levels of DNA methylation and DNMT1 mRNA were significantly decreased in the patients with anti-CCP antibody compared with those without (p=0.005, 95% CI=-0.7333 to -0.1373 and p=0.003, 95% CI=-0.0071 to -0.0022, respectively). The differences in the methylation level and expressions of DNMT1 and MBD2 were not significant between the patients treated with and without anti-TNFα biological agents (Enbrel or Humira).This study demonstrated that the RA patients have a significantly lower level of DNA methylation than the controls. Moreover, RA patients have higher expressions of DNMT1 and MBD2 mRNA. The anti-TNFα biological agents do not seem to affect DNA methylation and mRNA expressions of DNMT1 and MBD2 in RA patients.

The objective of this study was to investigate the associations between suppressor of cytokine signaling 1 (SOCS1) mRNA expression and SOCS1 polymorphisms with the development of rheumatoid arthritis (RA). One hundred and eighty-one patients with RA and 96 healthy controls were enrolled in this study. The SOCS1 mRNA level in peripheral blood mononuclear cells (PBMCs) was detected by quantitative real-time polymerase chain reaction. SOCS1 polymorphisms were determined by the polymerase chain reaction/restriction fragment length polymorphism method. We found that the expression of SOCS1 mRNA in PBMCs was significantly greater in patients with RA than in healthy controls. There were no significant differences in the expression of SOCS1 mRNA among patients with different disease activities. The increment in SOCS1 mRNA after stimulation with various cytokines was slightly lower in the patients with RA than in the healthy controls. This study also demonstrated that the SOCS1 polymorphisms were not associated with susceptibility to RA. In conclusion, the expression of SOCS1 mRNA in PBMCs is higher in patients with RA than in healthy controls. The increment in SOCS1 mRNA expression in PBMCs after stimulation with different cytokines seems to be lower in patients with RA than in healthy controls.

To investigate the role ofIκBαpromoter polymorphisms in the development of Behçet’s disease, eighty-six patients with Behçet's disease and 120 healthy controls were enrolled in this study. TheIκBα; -881A/G, -826C/T, -550A/T, -519C/T, and -297C/T polymorphisms were measured by the method of polymerase chain reaction/ restriction fragment length polymorphism. This study demonstrated that the genotype frequencies ofIκBα-826C/T and -826T/T were significantly higher in the patients with Behçet's disease than in the controls. Both in the dominant and in the recessive models, the patients with Behçet's disease have higher frequencies of the IκBα -826T containing genotype than the controls. The allele frequency ofIκBα-826T was significantly increased in the patients with Behçet’s disease. The frequencies of theIκBα-881A -826T -550A -519C -297C andIκBα-881A -826T -550A -519T -297C haplotypes were significantly higher in the patients with Behçet–s disease than in the controls. In contrast, the haplotype frequency ofIκBα-881A -826C -550A -519C -297C in the patients with Behçet’s disease was significantly decreased. This study also revealed that the Behçet’s disease patients with IκBα -826T/T have higher prevalence of skin lesions than those without IκBα -826T/T. In summary, the IκBα -826T allele,IκBα-881A -826T -550A -519C -297C andIκBα-881A -826T -550A -519T -297C haplotypes might be associated with susceptibility to Behçet’s disease. TheIκBα-826T/T genotype was related to the development of skin lesions in the patients with Behçet's disease.

We implemented a nationwide population-based study in Taiwan to compare the physical and biochemical parameters, diet and exercise lifestyles, and prevalences of diabetes, hyperlipidemia, and hypertension between males and females, and to clarify the determinants of diabetes, hyperlipidemia, and hypertension in Taiwan. In this cross-sectional study, 7,578 subjects were selected from the general population by stratified random sampling for the Surveillance of Taiwanese Civil Health in 2002. Blood samples were taken and information on body composition, demographics, exercise and dietary habits, and medical and drug histories were obtained from structured interviews administered by well-trained interviewers. A total of 6,600 subjects (87.1%), aged 15.6–95.0 years old, completed the survey. The overall prevalences of diabetes, hyperlipidemia, and hypertension were 9.9%, 22.8%, and 15.7%, respectively, and hyperlipidemia (27.0%) and hypertension (19.2%) were more prevalent in males. Males were more likely to have high-fat and high-cholesterol diets, compared with females. Although there were differences in the prevalences of hyperlipidemia and hypertension between the sexes, adjusted logistic regression analysis demonstrated little contribution of diet and exercise habits to the risks of diabetes, hyperlipidemia, or hypertension after adjusting for age, sex, waist-to-hip ratio, serum blood sugar levels, cholesterol, triglycerides, apolipoprotein A1, apolipoprotein B, glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, creatinine, uric acid, and blood pressure.

The relationship between cigarette smoking and cell damage is complicated, particularly considering the role of oxidative stress. The aim of this study was to identify the relationships among plasma nicotine metabolites, lipophilic antioxidants, and metabolic parameters in smokers and nonsmokers. This cross-sectional study recruited 100 subjects who visited the Department of Family Medicine at Kaohsiung Medical University Hospital. Excluding 14 ineligible cases, 46 smokers and 40 non-smokers were enrolled. Plasma nicotine metabolites, lipophilic antioxidants (including retinol, lycopene, alpha-carotene, beta-carotene, delta-tocopherol, gamma-tocopherol and alpha-tocopherol), related metabolic parameters, and body composition (including height, weight, body mass index, body fat, and waist circumference) were examined by comparison of means, correlations and regressions. Significant correlations among nicotine metabolites, age, sex, body composition and plasma lipophilic antioxidants were noted. Nicotine metabolites, age, body height and body weight were closely associated with plasma antioxidant levels (p0.05) in multiple linear regression. The levels of alpha-carotene, beta-carotene, gamma-tocopherol and lycopene were lower in smokers than in non-smokers (p0.01). The plasma level of high-sensitivity C-reactive protein (hsCRP), which is a marker for high cardiovascular risk, was higher in smokers than in non-smokers (p = 0.003). We conclude that the lower plasma antioxidant levels and the higher level of hsCRP in smokers may lead to decreased protective efficacy compared with non-smokers. Further studies are warranted to support our hypothesis.

Most researchers believe that the peroxisome proliferative activated receptor gamma (PPARgamma-2) and bone morphogenetic protein receptor type II (BMPR2) play important roles in steroid-induced osteonecrosis (ON). However, the molecular mechanism of this process is still unclear. Recent studies indicate that steroid treatments cause adipocyte formation due to differentiation of mesenchymal stem cells, which then prevents osteoblast formation. This study examined PPARgamma-2, bone morphogenetic protein 2 (BMP2), and BMPR2 in patients with systemic lupus erythromatosus (SLE) who eventually developed ON after prolonged steroid treatment. The subjects of this experiment included 220 SLE patients who had undergone steroid treatment for at least 2 years. Fifty-five of the 220 patients were ON patients, and 165 were non-ON patients. Real-time PCR was performed to analyze the expression of the PPARgamma-2, BMP2, and BMPR2 mRNA in the peripheral blood of these patients. The results indicated that the expression of PPARgamma-2 mRNA increased 37% in the ON patients' peripheral blood, but the expression of BMPR2 mRNA decreased 57%. The average expression of the PPARgamma-2 mRNA in the ON patients was significantly higher than that in the non-ON patients (p = 0.044). Conversely, the expression of BMPR2 mRNA was significantly lower than that in non-ON patients (p = 0.036), but the expression of BMP2 mRNA did not significantly differ. This study demonstrated that the PPARgamma-2 and BMPR2 have important roles in the ON process after prolonged steroid administration in SLE patients; however, the detailed molecular mechanisms of this process require further study.

To investigate the association of IkBα promoter polymorphisms with the development of primary Sjogren’s syndrome in Taiwan, 98 patients with primary Sjogren’s syndrome and 110 unrelated healthy controls were enrolled in this study. The IκBα −881 A/G, IκBα −826 C/T, IκBα −550 A/T, IκBα −519 C/T, and IκBα −297 C/T polymorphisms were determined by the methods of polymerase chain reaction/restriction fragment length polymorphism. This study demonstrated that the genotype frequencies of IκBα −826 C/T and IκBα −826 T/T, in comparison with that of IκBα −826 C/C, were significantly higher in the patients with primary Sjogren’s syndrome than in the controls. The allele frequency of IκBα −881 G was significantly decreased in the patients with primary Sjogren’s syndrome compared with that of the controls. In contrast, the allele frequency of IκBα −826 T was significantly higher in the patients with primary Sjogren’s syndrome than in the controls. The similar findings could also be found in the allele carriage frequencies. The patients with primary Sjogren’s syndrome had lower allele carriage frequencies of IκBα −881 G and IκBα −826 C, and a higher allele carriage frequency of IκBα −826 T. We also found that the estimated haplotype frequency of IκBα −881A-826T-550A-519C-297C was significantly increased in the patients with primary Sjogren’s syndrome in comparison with that of the controls. This study demonstrated that the IkBα −826T allele and IkBα −881A-826T-550A-519C-297C haplotype were associated with susceptibility to primary Sjogren’s syndrome in Taiwan. However, these findings may not be disease-specific but may be related to inflammatory responses.

Background. Cross-sectional studies investigated the impact of renin-angiotensin system (RAS) gene polymorphism on left ventricular mass index (LVMI) with conflicting results. We conducted a longitudinal study to investigate the influence of the angiotensin-converting enzyme (ACE) insertion/deletion (I/D) and angiotensinogen (AGT) M235T and angiotensin II type 1 receptor (AT1R) A1166C gene polymorphisms on the LVMI and geometry. Methods. Of 1500 people screened, 110 nondiabetic normotensive elderly Chinese persons were recruited and received echocardiography at baseline and at the 2nd and 4th year follow-up. No participants had a history of organic heart disease or chronic medication. The gene polymorphisms were analyzed by using polymerase chain reaction. Results. Participant age was 71.9 6 3.9 years (range 60‐81 years). The prevalence of concentric remodeling, eccentric hypertrophy, and concentric hypertrophy was significantly increased as well as LVMI after 4 years (all p , .05). These changes and the magnitude of LVMI increase were significantly higher in participants carrying the ACE D allele than nonD-allele carriers (all p , .05). This association was still significant in multivariate analyses (p � .02). A similar analysis showed a borderline significance in the AT1R but not in the AGT gene polymorphism. Conclusions. This longitudinal study showed that the aging process was associated with an increase of LVMI and changes of geometry. The RAS gene polymorphism, especially the ACE D allele, might modulate these changes in the Chinese population. This provides further knowledge that is essential in the assessment of cardiac disease and the determination of the left ventricular structure in older persons.

OBJECTIVES: To test the association between fasting glucose level and left ventricular mass (LVM) and left ventricular hypertrophy (LVH) in people aged 60 and older. DESIGN: Population-based prospective study with 4-year follow-up. SETTING: Department of Internal Medicine and Family Medicine, Kaohsiung Medical University (KMU), Chung-Ho Memorial Hospital, and Graduate Institute of Medicine and Public Health, KMU. PARTICIPANTS: Of 1,500 people screened, 105 without symptoms or signs of diabetes mellitus, hypertension, or cardiovascular disease were recruited from senior activity centers in Kaohsiung city. MEASUREMENTS: All received two-dimensional echocardiography and fasting glucose examination at baseline and at 2- and 4-year follow-up. LVH was defined as a LVM index (LVMI) greater than 122.4 g/m2 or 51 g/m2.7. RESULTS: Age ranged from 60 to 81 (mean 71.7±3.9). Baseline glucose ranged from 83 to 118 mg/dL (mean 99.7±7.9 mg/dL). LVMI was significantly higher at the 4-year follow-up (97.5±24.9 vs 104.5±27.5 g/m2 and 44.2±12.1 vs 47.2±13.4 g/m2.7, both P

To investigate the role of SLC 11A1 polymorphisms in the development of reactive arthritis, 91 patients with reactive arthritis and 163 healthy controls were enrolled in this study. The SLC 11A1 polymorphisms were determined by the method of polymerase chain reaction/restriction fragment length polymorphism. The genotype distributions of SLC 11A1 274, 823, 1703, and 1729+ 55 del 4 were significantly different between the patients with reactive arthritis and controls. The genotype frequency of SLC 11A1 274C/C was significantly decreased in the patients with reactive arthritis when compared with that of the controls. In contrast, the SLC 11A1 274C/T showed a significant association with reactive arthritis. The patients with reactive arthritis have a significantly higher frequency of SLC 11A1 823C/C than the controls. However, SLC 11A1 823T/T was resistant to the development of reactive arthritis. The allele frequencies of SLC 11A1 274T and 823C were significantly increased in the patients with reactive arthritis in comparison with those of the controls, independent of HLA-B27. On the contrary, the allele frequencies of SLC 11A1 274C and 823T were significantly decreased in the patients with reactive arthritis. The estimated haplotype frequency of SLC 11A1 274C 823T 1703G 1729+55del 4 TGTG+ was significantly decreased in the patients with reactive arthritis when compared with that of the controls. In contrast, the estimated haplotype frequency of SLC 11A1 274T 823C 1703G 1729+55 del 4 TGTG+ was significantly increased in the patients with reactive arthritis. This study shows that the SLC 11A1 274T and 823C alleles are associated with susceptibility to reactive arthritis independently of HLA-B27 in Taiwan. The SLC 11A1 274T 823C 1703G 1729+55 del 4 TGTG+ haplotype is associated with the development of reactive arthritis in Taiwan. In contrast, the SLC 11A1 274C 823T 1703G 1729+55 del 4 TGTG+ haplotype may be a protective factor.

Our previous studies showed that colorectal tumor has high interleukin-4 receptor α (IL-4Rα) expression, whereas adjacent normal tissue has low or no IL-4Rα expression. We also observed that human atherosclerotic plaque-specific peptide-1 (AP1) can specifically target to IL-4Rα. In this study, we investigated the therapeutic efficacy and systemic toxicity of AP1-conjuagted liposomal doxorubicin. AP1 bound more strongly to and was more efficiently internalized into IL-4Rα-overexpressing CT26 cells than CT26 control cells. Selective cytotoxicity experiment revealed that AP1-conjugated liposomal doxorubicin preferentially killed IL-4Rα-overexpressing CT26 cells. AP1-conjugated liposomal doxorubicin administered intravenously into mice produced significant inhibition of tumor growth and showed decreased cardiotoxicity of doxorubicin. These results indicated that AP1-conjugated liposomal doxorubicin has a potent and selective anticancer potential against IL-4Rα-overexpressing colorectal cancer cells, thus providing a model for targeted anticancer therapy.

To investigate the role of programmed cell death-1 (PD-1) gene polymorphisms in the development of systemic lupus erythematosus (SLE) in Taiwan, 109 patients with SLE and 100 healthy controls were enrolled in this study. The PD-1 gene polymorphisms were determined by the method of polymerase chain reaction/restriction fragment length polymorphism. This study showed that the genotype distributions of PD-1 7209 C/T polymorphisms were significantly different between the patients with SLE and controls (P=0.002, Pc=0.018). The frequencies of the PD-1 7209 C/C genotype and PD-1 7209 C allele were significantly higher in the patients with SLE than those of the controls (P=0.001, OR=2.6, 95% CI=1.5-4.6, and P=0.002, OR=2.1, 95% CI=1.3-3.4, Pc=0.018, respectively). Moreover, the association of PD-1 7209 C with susceptibility to SLE was independent of the PD-1 ligand. This study also showed that the PD-1-536 A 7146 G 7209 C 7499 G haplotype was associated with the development of SLE in Taiwan.

Objectives To investigate the role of inhibitor of kB-like ( IkBL ) gene polymorphisms in the pathogenesis of rheumatoid arthritis (RA) in Taiwan. Methods One hundred and twenty-nine patients with RA and 110 healthy controls were enrolled in this study. Polymerase chain reaction (PCR)/direct sequencing was used to determine the polymorphisms of IkBL −421 8T/9T, −324 C/G, −262 A/G, and −62 A/T. PCR/restriction fragment length polymorphism was used to determine the IkBL +738 T/C polymorphisms. Results The genotype distribution of IkBL −421 was significantly different between DR4 (+) RA patients and DR4 (+) controls ( p = 0.02). The allele frequency of IkBL −421 8T was significantly higher in DR4 (+) RA patients than in DR4 (+) controls ( p = 0.004, OR = 7.2, 95% CI = 1.7–29.2). The allele carriage frequency of IkBL −421 8T also tended to be increased in DR4 (+) RA patients in comparison with DR4(+) controls ( p = 0.07, OR = 14.6, 95% CI = 1.4–147.0). We also found that the allele frequency of IkBL −62 T was significantly higher in RA patients than in controls ( p = 0.04, OR = 1.5, 95% CI = 1.1–2.1). The allele carriage frequency of IkBL −62 T tended to be increased in RA patients ( p = 0.08, OR = 1.7, 95% CI = 1.0–3.0). The estimated haplotype frequency of IkBL −421 8T/−62 T tended to be increased in RA patients compared with controls ( p = 0.07, OR = 1.4, 95% CI = 1.0–2.0). Conclusion The IkBL −62 T may be associated with the development of RA in Taiwan. The IkBL −421 8T may also be related to susceptibility to RA in HLA-DR4 (+) individuals. This study shows that the estimated haplotype IkBL −421 8T/−62 T tends to be associated with susceptibility to RA in Taiwan.

Invitrofolate deficiency is associated with S phase accumulation and apoptosis in various cell types. To investigate the role of p53 and two apoptosis-related molecules, bcl-2 and Fas antigen (Apo-1, CD95), in the mechanism whereby folate-deficient lymphocytes accumulate and undergo apoptosis in the S phase, normal human peripheral blood lymphocytes were cultured for 3–9 d in control medium or in specially ordered and formulated HAM’ F-10 medium lacking folic acid, thymidine and hypoxanthine. Cells were stimulated with phytohaemagglutinin for the final 72 h prior to harvesting. The results indicate that p53 expression was downregulated in folate-deficient lymphocytes when compared with the control lymphocytes during the relevant period of S phase accumulation and apoptosis. In addition, folate deficiency was also found to downregulate IL-2, Fas antigen and bcl-2 expression, in terms of either mRNA or protein levels. The downregulation of Fas antigen suggests that folate deficiency-induced apoptosis probably does not occur via the Fas pathway. As IL-2 is a known inducer of bcl-2, and the downregulation of bcl-2 induces apoptosis, the downregulation of IL-2 and bcl-2 is suggested to play an important role in apoptosis. The complete rescue of folate-deficient lymphocytes from apoptosis was achieved by folic acid, thymidine or hypoxanthine alone or thymidine and hypoxanthine in combination. These results suggest that IL-2 depletion by folate deficiency in lymphocytes reduces the bcl-2 level, thereby triggering deoxynucleoside triphosphate pool imbalance and p53-independent apoptosis.

Objective Homocystinemia is an important independent risk factor for atherosclerosis. Inflammatory cytokines play key roles in the development of atherogenesis. This study investigated the effect of homocysteine on inflammatory cytokine expression. Methods Human monocytes were treated in vitro with a variety of dl -homocysteine concentrations that ranged from physiologic concentration to higher than pathophysiologic concentration, and we analyzed their expressions of inflammatory cytokines, including tumor necrosis factor-α, interleukin-1β, interleukin-6, interleukin-8, interleukin-12, and migration inhibitory factor. Results dl -homocysteine at a marginal physiologic concentration of 2 μg/mL (15 μM) activated monocytes. In addition, dl -homocysteine at the pathophysiologic dose of 25 μg/mL (185 μM) induced mRNA and protein expressions of inflammatory cytokines tumor necrosis factor-α, IL-1β, interleukin-6, interleukin-8, and interleukin-12. Moreover, at the larger dose of 50 μg/mL (370 μM) dl -homocysteine decreased expression of migration inhibitory factor at the mRNA and protein levels. Conclusion These findings suggest that homocysteine may contribute to the initiation and progression of vascular disease by activating monocytes, resulting in the secretion of cytokines that amplify the inflammatory response.

Oxidative stress triggered by septic insult may be the major cause of multiple organ dysfunction syndrome (MODS) in intensive unit care patients. The inducible form of heme oxygenase-1 (HO-1) can be induced by cytokines, lipopolysaccharide, and reactive oxygen species during sepsis. These facts raise the question of whether the expression of HO-1 in leukocytes can indicate the level of oxidative stress of multiple organs in sepsis. Clinical peritonitis was simulated in an animal model by cecal ligation and puncture (CLP). The level of oxidative stress was examined by plasma lipid peroxidation (LPO). Liver function was analyzed by plasma aspartate aminotransferase, alanine aminotransferase, total bilirubin, and direct bilirubin. Lung function was evaluated by severity of edema. Renal function was measured by blood urea nitrogen and creatinine. The correlation between early HO-1 induction and LPO level or organ functional indicators of the same rat at late sepsis was analyzed by linear regression. The results showed that the protein content of HO-1 increased at 9 h after CLP, whereas expression of HO-1 mRNA in leukocytes was significantly increased (P < 0.01) at 6 h after CLP. Plasma level of LPO and the indices of hepatic, pulmonary, and renal function were significantly increased at 18 h after CLP. Moreover, highly negative correlations were observed between HO-1 mRNA expression at 6 h after CLP and level of LPO or severity of hepatic/renal dysfunction at 18 h after CLP. These results suggest that early HO-1 mRNA expression in leukocytes may represent oxidative stress and may predict the severity of liver and renal dysfunction during sepsis.

Eugenosedin-A has been demonstrated to possess alpha/beta-adrenoceptor and serotonergic receptor blocking activities. We have investigated by what mechanisms eugenosedin-A prevents lipopolysaccharide (LPS)-induced hypotension, vascular hyporeactivity, hyperglycaemia, oxidative injury or inflammatory cytokines formation in rats. Intravenous administration of eugenosedin-A, trazodone, yohimbine (1 mg kg(-1)), aminoguanidine or ascorbic acid (15 mg kg(-1)) normalized LPS (10 mg kg(-1))-induced hypotension. Pretreatment with eugenosedin-A or the other agents 30 min before LPS injection reduced aortic hyporeactivity. LPS-induced increases in plasma interleukin-1beta (IL-beta), IL-6, interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha) and blood glucose levels were significantly inhibited by eugenosedin-A (1 mg kg(-1), i. v.). The same dose of trazodone, a chloropiperazinylbenzene-type antidepressant, and yohimbine, an alpha(2)-adrenoceptor antagonist, reduced IL-1beta and TNF-alpha, but it could not inhibit hyperglycaemia. Aminoguanidine, an inducible nitric oxide synthase (iNOS) inhibitor, and ascorbic acid, an antioxidant, decreased IL-1beta, TNF-alpha contents and hyperglycaemia. Eugenosedin-A and the other agents inhibited Fe(2+)-ascorbic acid-induced peroxidation in rat cortex, indicating that those agents had antioxidant effects, with the exception of aminoguanidine. In free radical scavenged experiments, eugenosedin-A and ascorbic acid eliminated peroxyl radicals. All test agents inhibited the LPS-induced increase of malondialdehyde (MDA) content in rat brain homogenates. When mice were administered an intraperitoneal injection of LPS alone, mortality occurred from 4 to 16 h, after which time all were dead. However, eugenosedin-A significantly prolonged the survival time after LPS injection, suggesting that eugenosedin-A protected against LPS-induced cardiovascular dysfunction, hyperglycaemia, tissue injury and inflammatory cytokine production. This was attributable mainly to the antioxidant and peroxyl radical scavenged effects of eugenosedin-A, and which may be, at least in part, due to its blockade on alpha/beta-adrenergic and serotonergic receptors.

The aim of this study was to develop a microdialysis-graphite furnace atomic absorption spectroscopy (MD-GFAAS) for monitoring dynamic changes of extracellular magnesium (Mg) and zinc (Zn) in the cortex of gerbils subjected to focal cerebral ischemia, that had been produced in anesthetized gerbils by occlusion of the right middle cerebral artery.Two microdialysis probes were inserted into both sides of the cortex to simultaneously collect dialysates during cerebral ischemia. Dynamic changes in these analytes, on ipsilateral and contralateral sides of the brain, were assayed by MD-GFAAS. Optimal conditions and analytical precision of GFAAS were studied in the present assay.The present study demonstrated significant decreases in Mg (65% of baseline) and zinc (74% of baseline) maintained their levels within 3 h on the ipsilateral side of cortex during cerebral ischemia. Slight changes of Mg and Zn on the contralateral sides were also observed.The derangement of extracellular Mg and Zn could be important in the progression of cell injury and may be associated with cerebral ischemia insult.

The major objective of the present study was to identify biologic and behavioral risk factors of cardiovascular disease (CVD) in the elderly population in Taiwan. It is hypothesized that the selected risk factors are significantly associated with the prevalence of CVD. Data came from a nationwide geriatric survey in 1991. Stratified proportional sampling was used to recruit 2,600 subjects. These were evaluated by family physicians working for the Departments of Family Medicine at four medical centers in four major cities in Taiwan. Univariate and multivariate logistic regression analyses were used to examine the associations between risk factors and the prevalence of CVD. The prevalence of CVD was 38.31%. Patients with CVD consistently had higher values for each selected risk factor except high-density lipoprotein-cholesterol (HDL-C) and glucose concentrations. The findings also indicated that hypertension, hypertriglyceridemia, low HDL-C concentration, ex-drinking status, and overweight were significantly associated with the prevalence of CVD among the elderly in Taiwan. The findings not only confirm the risk factors for CVD, but also invite more attention to be given to the importance of biologic and behavioral risk factors in CVD.

Objective:As there is little understanding of disability processes in Taiwanese elders, the present study aimed to identify medical predictors of the incidence of activities of daily living (ADL) disability. Method:A total of 903 subjects were studied over a 4-year period (1994–1998). The measurement instrument was the Chinese-version Multidimensional Functional Assessment Questionnaire (CMFAQ). Only the subjects free of ADL disability at baseline were included in separate logistic regression models to predict disability in physical and instrumental ADL. Results:The hypothesised associations between chronic conditions and future functional disability were cross-nationally confirmed. Arthritis, diabetes and hypertension were significantly predictive of the onset of physical ADL (PADL) disability; diabetes was the only medical predictor of instrumental ADL (IADL) disability. Age and educational level were significant predictors of PADL and IADL. Conclusions:Prevention programs for chronic disease need to pay more attention not only to the disabled population, but also to secondary prevention among those elders who have higher medical risk of developing disability.

Objective. The expression of autoimmunity in mice deficient in programmed death 1 (PD-1) suggests that PD-1 is a candidate gene involved in the development of human autoimmune diseases such as systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). We therefore tested the potential association between PD-1 and the development of SLE and RA by conducting case–control genetic-association studies. Methods. Ninety-eight SLE patients, 84 RA patients, and sex-matched control subjects for each disease group were recruited and genotyped for a single-nucleotide polymorphism, C+872T, in the human PD-1 gene. The significance of the association of the PD-1 gene with SLE or with RA was analyzed by statistical tests for the difference in genotype distribution between disease and control groups. Results. The human PD-1 gene was found to be significantly associated with disease development in RA patients, but not SLE patients. The risk of RA development appeared to be significantly increased by carriage of the T allele (odds ratio 3.32, P < 0.0001) or the C/T genotype (odds ratio 3.52, P < 0.00005). Conclusion. The PD-1 gene is significantly associated with RA susceptibility, suggesting the possibility that PD-1 may contribute to the pathogenesis of RA.

Objectives: To investigate the association of tumor necrosis factor (TNF) microsatellite alleles with the pathogenesis of rheumatoid arthritis (RA) in Taiwan. Methods: The TNF a, b, c, d, and e microsatellites were determined in 112 patients with RA and 99 healthy controls by using polymerase chain reaction (PCR) and electrophoresis with sequencing gel. All of these patients and controls had known HLA-DR genotypes and TNF-308 polymorphisms. Results: The phenotypic frequency of TNFa9 was significantly higher in DR4(−) RA patients than in DR4(−) controls. However, the phenotypic frequency of TNFb6 was significantly higher in RA patients than in controls in the presence of HLA-DR4. The phenotypic frequency of TNFa3-e1 was significantly lower in DR4(+) RA patients than in DR4(+) controls, while a negative linkage disequilibrium was noted between TNFa3-e1 and HLA-DR4. TNF microsatellite alleles were not related to the prevalences of bone erosion, rheumatoid nodule, sicca syndrome, pulmonary fibrosis, and seropositivity of rheumatoid factor (RF) in patients with RA. Conclusion: The associations of TNF microsatellites with the susceptibility to RA in Taiwan are not completely independent of the HLA-DR associations. The association of TNFb6 with the susceptibility to RA depends on the presence of HLA-DR4, and the correlation of TNFa9 to RA depends on the absence of HLA-DR4. The negative association of TNFa3-e1 with RA may be secondary to the negative linkage disequilibrium between TNFa3-e1 and HLA-DR4. Moreover, TNFb6 and HLA-DR4 have a synergistic effect on the susceptibility to RA. TNFa3-e1 and TNF-308A have a synergistic effect on preventing from RA. The TNF microsatellite alleles are not related to the clinical manifestations and severity of RA patients in Taiwan.

Bloodletting acupuncture is one of the most classic methods of acupuncture therapy, and is still popularly used to treat acute lumbar sprain in the oriental world. However, most physiciants in the western world are not familiar with bloodletting acupuncture, though they may know ordinary acupuncture well. Furthermore based on the literature reviewed, there have been few studies which have investigated the effect of bloodletting acupuncture upon acute lumbar sprain. In this study, we tried to determine if bloodletting acupuncture is effective for acute lumbar sprain. In total, twelve patients were enrolled for analysis. Five patients treated with ordinary acupuncture upon the contralateral SI-3 (Hou-Hsi) point alone. Seven patients were first treated with bloodletting acupuncture to the engorged vein around the ipisilateral Bl-40 (Wei-Chung), and then followed by ordinary acupuncture upon the contralateral SI-3. It was demonstrated that bloodletting acupuncture to the engorged vein around the ipisilateral Bl-40 followed by ordinary acupuncture upon the contralateral SI-3 had more pain relief than ordinary acupuncture upon the contralateral SI-3 alone (83 ± 23% vs. 44 ± 28%) (P < 0.01). And bloodletting acupuncture to the engorged vein around the ipisilateral Bl-40 decreased pain by 56 ± 23%, similar to that of ordinary acupuncture upon the contralateral SI-3 alone (44 ± 28%). These findings suggest that bloodletting acupuncture to the engorged vein around the ipisilateral Bl-40 (Wei-Chung) has a substantial contribution for treatment of acute lumbar sprain.

Background In Taiwan, distributions of body composition are usually generated for children, adolescent, and middle-aged groups. This study describes three anthropometric measures (i.e., height, weight, and body mass index [BMI]) of elderly persons in Taiwan. Methods The data was derived from a prospective cross-sectional, population-based geriatric survey funded by the National Department of Health in Taiwan in 1991. A total of 2,600 subjects were successfully evaluated. A descriptive analysis was performed to generate distributions of height, weight, and BMI of elderly persons. Based on these descriptive data, the prevalence of overweight and obesity was estimated, and a cross-national comparison on the prevalence of obesity was made. Results Age-group-specific means and standard deviations for height, weight, and BMI of Taiwanese elderly persons were presented. By gender, the elderly men were taller and heavier than the elderly women: however, the women had a higher mean value of BMI (kg/m2). The prevalence of overweight was 27.3% in men and 34.9% in women; the prevalence of obesity was 3.2% in men and 6.4% in women. The prevalence of obesity for the elderly men and women in Taiwan was lower than in Kuwait, Sweden, the United States, and the Native Americans, but higher than in Japan. Conclusions The comparison indicates cross-cultural differences existing in the prevalence of obesity, which may be due to aging, nutritional status, or environmental factors. For future research, the relationship of anthropometric data to socioeconomic status and behavioral factors will be examined.

The purpose of this study was to determine the correlation between the prevalence of hepatitis C virus (HCV) infection with duration of drug use and other risk factors among drug users. This survey covered 899 male drug users from Kaohsiung Narcotic Abstention Institute and Kaohsiung prison. The prevalence of positive anti-HCV was 67.2% among intravenous drug users (IVDU) and 14.7% among non-intravenous drug users (non-IVDU). Among intravenous (IV) drug users, age and duration of drug use were independently related to HCV seropositivity. Seroprevalence rate for HCV in the IVDU group increased with increasing duration of injection use within the first seven years of drug use. However, the steepest trajectory in seroprevalence of HCV infection occurred within the first four months. Due to the high rate of HCV infection among drug users, investigation of risk behaviors should be routine in such a group. A high frequency of HCV infection was also found among short-term injectors, which indicated that early risk reduction intervention was an important measure in moderating HCV infection.

Many autoimmune diseases have been reported to be associated with malignancy. Mixed connective tissue disease (MCTD), however, has rarely been associated with malignancy. Thymoma is one of the neoplasms often reported to be related to various immunological disorders. Among the types of thymoma defined by WHO, malignant thymoma (thymoma type C) is the one least reported to be associated with autoimmune disease. Here, we report a case of malignant thymoma with concurrent MCTD, which manifested with acrosclerosis, Raynaud's phenomenon, arthritis (synovitis), and a high titer of anti-ribonucleoprotein antibody.

Nitric oxide (NO) is a short-lived free radical that is secreted by pulmonary macrophages (Mø). An inducible isoform of NO synthase (iNOS) catalyses the production of NO and is activated by lipopolysaccharide and certain T-helper(h) 1 cytokines, including interferon-gamma and TNF-alpha. In the present study, iNOS+ interstitial cells were demonstrated in the alveolar wall of normal Lewis rat lung. Enzymatic digests of normal lung showed that approximately one third of pulmonary ED1+ interstitial Mø (IM) were iNOS+ and secreted modest amounts of NO without ex vivo stimulation, whereas normal alveolar macrophages (AM) were iNOS- and showed no basal NO secretion. When incubated with heat-killed Listeria monocytogenes (HKL) in vitro, AM secreted larger amounts of NO than did IM. Recombinant murine GM-CSF stimulated production of NO by AM but not by IM. However, when IM were costimulated with GM-CSF and IFN-gamma, they expressed a marked increase in NO production. Intratracheal challenge with HKL yielded decreased NO production by IM. We conclude that iNOS+ IM are present in normal rat lung, where they regulate the pulmonary cell-mediated immune response to antigen.