Kangfuxin alleviates ulcerative colitis in rats by inhibiting NF-κB p65 activation and regulating T lymphocyte subsets.

Objectives: Ulcerative colitis (UC) remains an enduring, idiopathic inflammatory bowel disease marked by persistent mucosal inflammation initiating from the rectum and extending in a proximal direction. An ethanol extract of Periplaneta americana L., namely Kangfuxin (KFX), has a significant historical presence in Traditional Chinese Medicine and has been broadly utilized in clinical practice for the treatment of injury. Here, we aimed to determine the effect of KFX on 2,4,6-trinitro'benzene sulfonic acid (TNBS)-induced UC in Sprague-Dawley rats. Materials and Methods: We established the UC model by TNBS/ethanol method. Then, the rats were subject to KFX (50, 100, 200 mg/kg/day) for 2 weeks by intragastric gavage. The body weight, disease activity index (DAI), colonic mucosal injury index (CMDI), and histopathological score were evaluated. The colonic tissue interleukin (IL)-1ß, IL-6, tumor necrosis factor-α (TNF-α), IL-10, transforming growth factor-1 (TGF-ß1), and epidermal growth factor (EGF) were determined by Elisa. To study T-lymphocyte subsets, flow cytometry was performed. In addition, the expression level of NF-κB p65 was evaluated by immunohistochemistry and western blot analysis. Results: Compared with the TNBS-triggered colitis rats, the treatment of rats with KFX significantly increased the body weight, and decreased DAI, CMDI, and histopathological score. Also, KFX elicited a reduction in the secretion of colonic pro-inflammatory cytokines, namely IL-1ß, IL-6, and TNF-α, concomitant with up-regulation of IL-10, TGF-ß1, and EGF levels. Upon KFX treatment, the CD3+CD4+/CD3+CD8+ ratio in the spleen decreased, while the CD3+CD8+ subset and the CD3+CD4+CD25+/CD3+CD4+ ratio demonstrated an increase. In addition, the expression of NF-κB p65 in the colon was decreased. Conclusion: KFX effectively suppresses TNBS-induced colitis by inhibiting the activation of NF-κB p65 and regulating the ratio of CD4+/CD8+.

The therapeutic effect of wasp venom (Vespa magnifica, Smith) and its effective part on rheumatoid arthritis fibroblast-like synoviocytes through modulating inflammation, redox homeostasis and ferroptosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Rheumatoid arthritis (RA) is a chronic inflammatory disease that is related to the aberrant proliferation of fibroblast-like synoviocytes (FLS). Wasp venom (WV, Vespa magnifica, Smith), an insect secretion, has been used to treat RA in Chinese Jingpo national minority's ancient prescription. However, the potential mechanisms haven't been clarified. AIM OF THE STUDY: The purposes of this paper were two-fold. First, to investigate which was the best anti-RA effective part of WV-I (molecular weight less than 3 kDa), WV-II (molecular weight 3-10 kDa) and WV-III (molecular weight more than 10 kDa) that were separated from WV. Second, to explore the underlying molecular mechanism of WV and WV-II that was best effective part in RA. MATERIALS AND METHODS: The wasps were electrically stimulated and the secretions were collected. WV-I, WV-II and WV-III were acquired by ultracentrifuge method according to molecular weight. Next, WV, WV-I, WV-II and WV-III were identified by HPLC. Functional annotation and pathway analysis of WV used to bioinformatics analysis. RNA-seq analyses were constructed to identify differentially expressed genes (DEGs). GO and KEGG pathway analyses were performed by Metascape database. STRING was used to analyze the PPI network from DEGs. Next, PPI network was visualized using Cytoscape that based on MCODE. The pivotal genes of PPI network and MCODE analysis were verified by qRT-PCR. Subsequently, MH7A cells were performed by MTT assay to evaluate the ability of inhibiting cell proliferation. Luciferase activity assay was conducted in HepG2/STAT1 or HepG2/STAT3 cells to assess STAT1/3 sensitivity of WV, WV-I, WV-II and WV-III. Additionally, interleukin (IL)-1ß and IL-6 expression levels were detected by ELISA kits. Intracellular thioredoxin reductase (TrxR) enzyme was evaluated by TrxR activity assay kit. ROS levels, lipid ROS levels and Mitochondrial membrane potential (MMP) were assessed by fluorescence probe. Cell apoptosis and MMP were measured by using flow cytometry. Furthermore, the key proteins of JAK/STAT signaling pathway, protein levels of TrxR and glutathione peroxidase 4 axis (GPX4) were examined by Western blotting assay. RESULTS: RNA-sequencing analysis of WV displayed be related to oxidation-reduction, inflammation and apoptosis. The data displayed that WV, WV-II and WV-III inhibited significantly cells proliferation in human MH7A cell line compared to WV-I treatment group, but WV-III had no significant suppressive effect on luciferase activity of STAT3 compared with IL-6-induced group. Combined with earlier reports that WV-III contained major allergens, we selected WV and WV-II further to study the mechanism of anti-RA. In addition, WV and WV-II decreased the level of IL-1ß and IL-6 in TNF-α-induced MH7A cells via inactivating of JAK/STAT signaling pathway. On the other hand, WV and WV-II down-regulated the TrxR activity to produce ROS and induce cell apoptosis. Furthermore, WV and WV-II could accumulate lipid ROS to induce GPX4-mediated ferroptosis. CONCLUSIONS: Taken together, the experimental results revealed that WV and WV-II were potential therapeutic agents for RA through modulating JAK/STAT signaling pathways, redox homeostasis and ferroptosis in MH7A cells. Of note, WV-II was an effective part and the predominant active monomer in WV-II will be further explored in the future.

Isolation and identification of the components in Cybister chinensis Motschulsky against inflammation and their mechanisms of action based on network pharmacology and molecular docking.

ETHNOPHARMACOLOGICAL RELEVANCE: Cybister chinensis Motschulsky belongs to the family Dytiscidae. As a traditional Chinese medicine, the insect is called Longshi in the folk and is commonly used to treat enuresis in children and frequent urination in the elderly. AIM OF THE STUDY: Inflammation is involved in chronic kidney disease. The previous study proved ethanol extract of C. chinensis exhibited anti-inflammation effects in the Doxorubicin-induced kidney disease. However, the material basis and their possible mechanism of the insect were still unclear. Thus, we aimed to separate the active compounds of the ethanol extract from C. chinensis and to investigate their possible mechanism of anti-inflammation by network pharmacology and molecular docking. MATERIALS AND METHODS: The insect was extracted with 75% ethanol to produce ethanol extracts and then were extracted by petroleum ether, ethyl acetate and n-butanol respectively. Silica gel column chromatography and preparative HPLC were applied to separate the compounds of the extract. The compounds were characterized and identified by NMR and mass. The compound associated genes were collected by BATMAN-TCM database and the inflammation associated genes were obtained through DigSee database. The protein-protein interaction (PPI) network was carried out via Search Tool for the Retrieval of Interacting Genes/Protein (STRING) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) target pathway analysis was performed in Database for Annotation, Visualization and Integrated Discovery (DAVID). The possible mechanism of compounds against inflammation was investigated by molecular docking. Finally, the anti-inflammatory effect of the representative compound was verified by the LPS-induced Raw 264.7 cell inflammatory model. TNF-α, IL-1ß and IL-6 of the cell supernatants were analyzed via using ELISA kits and the key proteins in JAK2/STAT3 signaling pathway were verified via the Western blot assays. RESULTS: Among crude extracts from C. chinensis, ethyl acetate extract showed the obvious anti-inflammatory effects. Nine compounds were isolated from ethyl acetate extract of Cybister chinensis for the first time, including benzoic acid (1), hydroxytyrosol (2), protocatechualdehyde (3), N-[2-(4-hydroxyphenyl)ethyl]acetamide (4), (2E)-3-phenylprop-2-enoic acid (5), 3-phenylpropionic acid (6), methyl 3,4-dihydroxybenzoate (7), 1,4-diphenyl butane-2,3-diol (8) and p-N,N-dimethylaminobenzaldehyde (9). After searching in the database, 1079 compound associated genes and 467 inflammation associated genes were found. The 137 common targets covered 77 signaling pathways, in which HIF-1 signaling pathway, TNF signaling pathway, influenza A, PI3K/Akt signaling pathway, NOD-like receptor signaling pathway, MAPK signaling pathway, Toll-like receptor signaling pathway and Jak-STAT signaling pathway were important for inflammation. Molecular docking studies showed compound 1, 4, 5, 6, 7 and 8 were the potential inhibitors of JAK2 protein. In addition, the in vitro test showed compound 5 reduced the expression of tumor necrosis factor (TNF)-α, interleukin (IL)-6 and IL-1ß in lipopolysaccharide (LPS)-stimulated RAW264.7 cells in a dose-dependent manner. Furthermore, it was found that compound 5 inhibited the expression of p-JAK2 and p-STAT3 in LPS-induced RAW264.7 cells in a dose-dependent manner. CONCLUSIONS: Based on the network pharmacology and molecular docking, the study suggested that C. chinensis could relieve the inflammation based on the multi-compounds and multi-pathways, which provided the foundation for the medicinal application of C. chinensis.

Kangfuxin Liquid Ameliorates Dextran Sulfate Sodium (DSS)-Induced Acute Ulcerative Colitis in Mice by Modulating Immune Response and Suppressing Inflammation.

BACKGROUND The aim of this study was to determine the effect of kangfuxin liquid (KFXL) on inflammatory response, and its underlying mechanism in treating acute ulcerative colitis (UC) in mice induced by dextran sulfate sodium (DSS). MATERIAL AND METHODS Mice were provided drinking water containing DSS (3%) for 7 days to induce acute enteritis. The mice were divided into 6 groups: a control group, a DSS-induced (vehicle) group, a sulfasalazine (SASP) group, and low-, medium-, and high-dose kangfuxin liquid groups. Disease activity index (DAI), colon mucosa damage index (CMDI), histopathological score (HS), and organ index were monitored daily. The levels of interleukin-1ß (IL-1ß), interleukin-10 (IL-10) in serum and interleukin-17 (IL-17) and epidermal growth factor (EGF) in colon tissue were assessed by enzyme-linked immunosorbent assay (ELISA). Flow cytometry was used to assess the changes of T lymphocyte subsets in spleens of mice to evaluate the therapeutic effect of drugs on acute UC in mice. RESULTS Different doses of kangfuxin liquid reduced the DAI, CMDI, and HS scores (P<0.01 or P<0.05) of acute UC mice, reduced the level of IL-1ß and IL-17 in serum, increased the expression of IL-10 in serum and EGF in colon tissue, increased the number of CD3⁺ T cells, and decreased the level of CD4⁺ T cells and the ratio of CD4⁺/CD8⁺. CONCLUSIONS Kangfuxin liquid has a therapeutic effect on DSS-induced acute UC in mice, and its mechanism of action may be associated with regulating immune function and reducing intestinal inflammatory response.

Madelung disease: A case report.

BACKGROUND: Madelung disease (MD) is a rare disorder of fat metabolism, resulting in diffuse, symmetrical and painless deposition of adipose tissue in subcutaneous superficial fascial space and/or deep fascia space of the head, neck and shoulders, etc. CASE SUMMARY: We report a case of MD accompanied by type 2 diabetes in a 61-year-old Chinese male. The patient presented with progressive fat deposition over the mandible, neck, abdomen and elbows. He had a history of smoking and alcohol abuse. Excessive fat deposition was seen in the mandible, elbows and the abdominal area of the patient by ultrasonic examination. Computed tomography showed diffuse and marked soft masses (fat density) in the subcutaneous superficial fascia space of the neck. The patient was diagnosed with MD. He was advised to abstain from alcohol and was followed up regularly. CONCLUSION: This report discusses the pathogenesis, diagnosis and treatment of MD, and raises the clinician's awareness of this disease.

Wasp Venom Possesses Potential Therapeutic Effect in Experimental Models of Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is an autoimmune disease. Wasp venom (WV), which is considered as a traditional folk medicine in Jingpo nationality in Yunnan, China, relieves rheumatoid arthritis. The current study aimed to investigate the effect of wasp venom ameliorating rheumatoid arthritis symptoms in experimental rats. We established a model of type II collagen- (CII-) induced arthritis (CIA) in SD rats and examined the inhibition of inflammation and autoimmune response. The antiarthritic effects of WV were evaluated through the paw swelling, and histopathological score and histopathology changes of the affected paw were assessed. The anti-inflammation effects were assayed by the level of IL-6, TNF-α, IL-1ß, and the number of inflammatory cells in peripheral blood. The alteration of the T cell subset ratio in the spleen of rats was detected by flow cytometry, and at the same time, the viscera index and immune serum globulin levels were evaluated. The results suggested that various doses of WV (0.125, 0.25, and 0.5 mg/kg) significantly alleviated paw swelling and arthritis score in CIA rats with the untreated control (P < 0.05). WV (0.25 and 0.5 mg/kg) relieved synovial tissue lesions of ankle joints and histopathology scores of synoviocyte hyperplasia and inflammatory cell infiltration with vehicle group (P < 0.05). Regarding immunological regulation, 0.5 mg/kg WV lowered the immune serum globulin levels (P < 0.05), and we further found that WV (0.5 mg/kg) suppressed the immune response of Th cells, while enhancing the functions of Tc cells and Treg cells in spleen cells markedly (P < 0.05). The immunosuppressive action of WV displayed was analogous to its inhibitory effect on IL-1ß, TNF-α, IL-8, IL-6, COX-2, and PGE2 levels in rat serum. In conclusion, these findings demonstrated that WV exhibited antiarthritic activity, which might be associated with their inhibitory effects on immunoregulation and anti-inflammatory action.

Research progress and perspective in metabolism and metabolomics of psoriasis / 中华医学杂志(英文版)

Psoriasis is considered a systemic disease associated with metabolic abnormalities, and it is important to understand the mechanisms by which metabolism affects pathophysiological processes both holistically and systematically. Metabolites are closely related to disease phenotypes, especially in systemic diseases under multifactorial modulation. The emergence of metabolomics has provided information regarding metabolite changes in lesions and circulation and deepened our understanding of the association between metabolic reprogramming and psoriasis. Metabolomics has great potential for the development of effective biomarkers for clinical diagnosis, therapeutic monitoring, prediction of the efficacy of psoriasis management, and further discovery of new metabolism-based therapeutic targets.

A Strategy for Quality Control of Vespa magnifica (Smith) Venom Based on HPLC Fingerprint Analysis and Multi-Component Separation Combined with Quantitative Analysis.

As a folk medicine of the Jingpo minority in Yunnan province, the venom of Vespa magnifica has been commonly used for the treatment of rheumatoid arthritis. Quality standardization of the wasp venom is a necessary step for its pharmaceutical research and development. To control the quality of the wasp venom, a method based on high-performance liquid chromatography (HPLC) was developed for chemical fingerprint analysis. In the chromatographic fingerprinting, chemometrics procedures, including similarity analysis (SA), hierarchical clustering analysis (HCA), and principal component analysis (PCA), were applied to classify 134 batches (S1-S134) of wasp venom from different origins. The HPLC fingerprint method displayed good precision (Relative standard deviation, RSD < 0.27%), stability (in 16 h, RSD < 0.34%), and repeatability (RSD < 1.00%). Simultaneously, four compounds (VMS1, VMS2, VMS3, and VMS4) in the wasp venom were purified and identified. VMS1 was 5-hydroxytryptamine, and the other compounds were three peptides that were sequenced as follows: Gly-Arg-Pro-Hyp-Gly-Phe-Ser-Pro-Phe-Arg-Ile-Asp-NH2 (VMS2), Ile-Asn-Leu-Lys-Ala-Ile-Ala-Ala-Leu-Ala-Lys-Lys-Leu-Leu-NH2 (VMS3), and Phe-Leu-Pro-Ile-Ile-Gly-Lys-Leu-Leu-Ser-Gly-Leu-Leu-NH2 (VMS4). The quantifications for these components were 110.2 mg/g, 26.9 mg/g, 216.3 mg/g, and 58.0 mg/g, respectively. The results of this work indicated that the combination of the chemical fingerprint and quantitative analysis offers a reasonable way to evaluate the quality of wasp venom.

Expression of proprotein convertase subtilisin/kexin type 9 in the plasma of patients with psoriasis and its effect on the activation of peripheral CD4+ T cells / 中华皮肤科杂志

Objective To investigate the role of proprotein convertase subtilisin/kexin type 9 (PCSK9) in the pathogenesis of psoriasis by detecting the level of PCSK9 in the plasma of patients with psoriasis and evaluating its effect on the secretion of interferon gamma (IFN-γ) and interleukin-17A (IL-17A) by peripheral CD4+ T cells.Methods Totally,30 outpatients with psoriasis vulgaris and 30 healthy volunteers (controls) were enrolled from Hospital for Skin Diseases,Chinese Academy of Medical Sciences between February 2016 and December 2017.Of the 30 patients,16 were males,and 14 were females.Their age varied from 18 to 66 years,and the course of disease ranged from 1 month to 15 years.Peripheral venous blood samples were obtained from the patients and controls,and the plasma and was performed to measure mRNA expression of PCSK9 in the PBMC,and enzyme-linked immunosorbent assay (ELISA) to determine the concentration of PCSK9 in the plasma.Peripheral CD4+ T cells were isolated from the PBMC by magnetic bead method,and divided into 2 groups to be co-cultured with (experiment group) or without PCSK9 protein (control group).After 24-hour treatment,ELISA was conducted to detect the levels of IFN-γ and IL-17A in the culture supernatant.Statistical analysis was carried out by using two-sample t test for the comparison between the two groups,and Pearson correlation analysis for analyzing correlations between the plasma level of PCSK9 and psoriasis area and severity index (PASI) score in the patients with psoriasis.Results PCSK9 mRNA expression was undetected in the PBMC of the patients with psoriasis and controls.The plasma level of PCSK9 was significantly higher in the patients with psoriasis ([243.58 ± 11.91] μg/L) than in the healthy controls ([199.74 ± 31.09] μg/L,t =5.761,P ＜ 0.001).After co-culture of the peripheral CD4+ T cells from patients with PCSK9 protein,the levels of IFN-γ and IL-17A both significantly increased ([6 150.00 ± 212.13] ng/L,[1 532.00 ± 11.31] ng/L,respectively) compared with the control group co-cultured without PCSK9 protein ([4 650.00 ± 212.13] ng/L,[698.5 ± 266.58] ng/L,respectively;t =7.071,4.418 respectively,both P ＜ 0.05).IFN-γand IL-17A were undetected in the culture supernatant of CD4+ T cells from the healthy controls in the experiment group or control group.Conclusion The plasma level of PCSK9 increases in patients with psoriasis,which may be involved in the pathogenesis of psoriasis by activating peripheral CD4+ T cells.

[Modified osteotomy of olecranon for the management of inter-condylar fracture of the humerus].

OBJECTIVE: To explore the surgical method and clinical outcome of modified osteotomy of olecranon for the treatment of inter-condylar fracture of humerus. METHODS: From May 2007 to December 2012, 32 patients of intercondylar fracture of humerus were treated surgically through the approach of modified osteotomy of olecranon. The patients were 21 males and 11 females with a mean age of 46.3 years (ranged 18 to 65 years). Nineteen fractures occurred on the right extremity and 13 on the left extremity. According to the AO classification, type C1 fracture was found in 7, C2 in 11 and C3 in 14. Five patients suffered from open fracture (Gustilo type Iin 3, type II in 2). Other fractures occurred in 6 patients and the primary injury of nerve occurred 6. The healing of the osteotomy was evaluated with physical examination and plain X-ray film, and the function of elbow was assessed according to Cassebaum scale. RESULTS: All the patients were followed from 9 months to 5 years(average, 1.9 years). All the osteotomies healed at 7.4 weeks averagely after operation, and no nonunion, delayed union, fracture of ulna olecranon were found. Two cases had little pain on the elbow, heterotopic ossification occurred in 2 cases and cutting bone block loosed in 1 case. The function of the elbow showed excellent in 19 cases, good in 8, fair in 4 and poor in 1. CONCLUSIONS: The use of the approach of modified olecranon osteotomy for surgical management of intercondylar fracture of humerus has some advantages, it provides satisfactory stability with simple technical procedures avoiding inter-articular invasion, and it facilitates rehabilitation exercises and providing good results with low complication rates.

Design and Evaluation of Virtual Reality-Based Therapy Games with Dual Focus on Therapeutic Relevance and User Experience for Children with Cerebral Palsy.

OBJECTIVE: Virtual reality (VR)-based therapy for motor rehabilitation of children with cerebral palsy (CP) is growing in prevalence. Although mainstream active videogames typically offer children an appealing user experience, they are not designed for therapeutic relevance. Conversely, rehabilitation-specific games often struggle to provide an immersive experience that sustains interest. This study aims to design and evaluate two VR-based therapy games for upper and lower limb rehabilitation and to evaluate their efficacy with dual focus on therapeutic relevance and user experience. MATERIALS AND METHODS: Three occupational therapists, three physiotherapists, and eight children (8-12 years old), with CP Level I-III on the Gross Motor Function Classification System, evaluated two games for the Microsoft(®) (Redmond, WA) Kinect™ for Windows and completed the System Usability Scale (SUS), Physical Activity Enjoyment Scale (PACES), and custom feedback questionnaires. RESULTS: Children and therapists unanimously agreed on the enjoyment and therapeutic value of the games. Median scores on the PACES were high (6.24±0.95 on the 7-point scale). Therapists considered the system to be of average usability (50th percentile on the SUS). The most prevalent usability issue was detection errors distinguishing the child's movements from the supporting therapist's. The ability to adjust difficulty settings and to focus on targeted goals (e.g., elbow/shoulder extension, weight shifting) was highly valued by therapists. CONCLUSIONS: Engaging both therapists and children in a user-centered design approach enabled the development of two VR-based therapy games for upper and lower limb rehabilitation that are dually (a) engaging to the child and (b) therapeutically relevant.

Inhibitory kinetics of DABT and DABPT as novel tyrosinase inhibitors.

4-Dimethylaminobenzaldehyde-thiosemicarbazone (DABT) and 4-dimethylaminobenzaldehyde-N-phenyl-thiosemicarbazone (DABPT) were synthesized and established by (1)H and (13)C NMR and mass spectrum. Both compounds were evaluated for their inhibition activities on mushroom tyrosinase and their anti-tyrosinase kinetics was investigated. The results showed that both compounds exhibited significant inhibitory effects on activity of monophenolase and diphenolase; DABT and DABPT decreased the steady-state rate with 1.54 µM and 1.78 µM as their IC50 values respectively. The inhibitory effects of diphenolase activity exhibited sharp in a dose-dependent manner and their IC50 values were estimated as 2.01 µM and 0.80 µM, respectively. Kinetic analysis showed that their inhibition mechanism was reversible. The inhibition type of DABT was mix-type with inhibition constants KI = 1.77 µM and KIS = 6.49 µM, while that of DABPT displays non-competitive with the inhibition constant KI = 0.77 µM.

Over-expression of AQP7 contributes to improve insulin resistance in adipocytes.

OBJECTIVE: Aquaglyceroporin 7 (AQP7) is required for efflux of glycerol from adipocytes. In this study, we aimed to analyze expression profiles of AQP7 in the different differentiation phases of adipocytes and to investigate the role of AQP7 in the insulin resistance of adipocytes. METHODS: 3T3-L1 pre-adipocyte cells were induced to be fully differentiated adipocytes and then insulin resistance was induced by Dexamethasone (DXM) or TNF-α. Adenovirus vector with over-expression AQP7 (Ad-AQP7) was constructed and transfected into adipocytes. The expression level of AQP7 and phosphorylated PKB (p-PKB) were measured. The glycerol released from adipocytes and glucose consuming rate were tested too. RESULTS: AQP7 expression was gradually up-regulated along with the differentiation processing of 3T3-L1 preadipocytes, which was consistent with the expression level of p-PKB. Dexamethasone down-regulated the expression of AQP7, p-PKB and the glycerol content in adipocytes. Over-expression of AQP7 by transfecting Ad-AQP7 to insulin resistant adipocytes restored the phosphorylation of PKB and attenuated the glycerol secretion and glucose consuming rate of adipocytes. CONCLUSIONS: AQP7 is down-regulated in adipocytes with insulin resistance. The over-expression of AQP7 contributes to improve insulin resistance in adipocytes, which is potentially correlated with the increased phosphorylation of PKB.

Detecting plasma Epstein-Barr virus DNA to diagnose postradiation nasopharyngeal skull base lesions in nasopharyngeal carcinoma patients: a prospective study / 癌症

The diagnosis of postradiation nasopharyngeal skull base lesions in petients with nasopharyngeal carcinoma (NPC) is still a tough problem in clinical practice. An early and accurate diagnosis is important for subsequent management. We prospectively evaluated the diagnostic value of plasma Epstein-Barr virus(EBV) DNA in detecting postradiation nasopharyngeal skull base lesions in NPC patients. From July 2006 to September 2010, 90 patients with postradiation NPC (34 women and 56 men; median age: 42 years) met the selection criteria and were recruited in this study. All postradiation nasopharyngeal skull base lesions were found in the latest magnetic resonance imaging (MRI) examinations before endoscopic surgery, and the nasopharyngeal cavity was normal under flexible nasopharyngoscopy. Plasma EBV DNA detection was performed within 2 weeks before endoscopic surgery. A total of 90 endoscopic operations were successfully performed without any postoperative complications. Recurrences confirmed by postoperative pathology were found in 30 patients. The specificity, positive and negative predictive values of plasma EBV DNA detection were better than those of MRI. In addition, combining plasma EBV DNA detection with MRI improved the specificity and positive predictive values of MRI. Plasma EBV DNA detection followed by MRI would help to diagnose recurrence whereas MRI was unable. These results indicate that plasma EBV DNA is an effective and feasible biomarker for detecting postradiation nasopharyngeal skull base lesions in NPC patients.

[Clinical analysis of histiocytic necrotizing lymphadenitis in 68 cases].

OBJECTIVE: To investigate and analyze the clinical features in patients with histiocytic necrotizing lymphadenitis (HNL). METHODS: A total of 68 HNL patients at our hospital between the years of 1999 to 2009 were enrolled. The clinical data were collected from the hospital records and the relevant literature was reviewed. RESULTS: HNL mainly affected young people with an average age of (19 ± 13) years old and a female-to-male ratio of 1:1. All patients had lymphadenectasis. And 95.6% patients (65 cases) were feverish and 36.8% patients (25 cases) had mild hepatosplenomegaly; 25.0% (17 cases) upper respiratory symptoms such as sore throat and cough; 14.7% (10 cases) skin rash in their history; 51.5% (35 cases) leucopenia; 25.0% (17 cases)hepatic dysfunctions; 72.1% (44/61) elevated erythrocyte sedimentation rate (ESR); 11.1% (6/54) positive antinuclear antibody (ANA). The final diagnosis of HNL was confirmed by pathological examination and immunohistochemical staining of biopsy specimens. And 34 (50.0%) patients received glucocorticoid for 2 weeks to 3 months. Seven (10.3%) patients relapsed in which glucocorticoid was effective. Of 6 patients with positive ANA, one case was complicated with systemic lupus erythematosus (SLE) and another case diagnosed with SLE at 2 years after HNL. CONCLUSIONS: The clinical manifestations of HNL lack specificity so that it can be easily misdiagnosed. While the etiology of HNL remains elusive, the histopathological examination of affected lymph nodes has contributed greatly to the final diagnoses of HNL. Glucocorticoid therapy is recommended for treatment. Generally, HNL has an excellent prognosis but it often relapses. It should be noted that HNL may coexist with SLE or evolve ultimately into SLE.

[Expression and role of peroxisome proliferator-activated receptor gamma and nuclear factor-kappaB in pulmonary fibrosis].

OBJECTIVE: To evaluate the expression of peroxisome proliferator-activated receptor gamma (PPARgamma) and nuclear factor-kappaB (NF-kappaB) in lungs in patients with pulmonary fibrosis, and to explore their effect on the pathogenesis of pulmonary fibrosis. METHODS: Immunohistochemical technology was performed to investigate the PPARgamma and NF-kappaB expression in lung specimens from 16 cases of pulmonary fibrosis and 10 cases of normal controls. RESULTS: The positive score of PPARgamma (0.35+/- 0.08) in fibrosis group was lower than that in control group (0.42+/-0.04, P<0.05). The positive score of NF-kappaB (0.51+/- 0.11) in fibrosis group was higher than that in control group (0.38+/-0.04, P<0.05). There was negative correlation between PPARgamma and NF-kappaB expression in fibrosis group (P<0.05). CONCLUSION: The decreased expression of PPARgamma and enhanced expression of NF-kappaB play a role in the pathogenesis of pulmonary fibrosis, which may provide a new idea for treating this disease.

[Expression of major histocompatibility complex class II antigens on lungs in rat with bleomycin-induced pulmonary fibrosis].

OBJECTIVE: To explore the expression levels of MHC II molecules and its regulator genes CIITA on bleomycin-induced pulmonary fibrosis in rats, and to investigate the underlying immunologic mechanisms of pulmonary fibrosis. METHODS: The rats were treated with either a single intratracheal bleomycin injection (fibrosis group) or a normal saline injection (control group). The pathologic changes of lung tissues stained with HE and Masson were observed, and the contents of hydroxyproline were detected on the 7th and 28th day respectively after bleomycin administration. The expression of MHC II molecules in the lung tissues was evaluated with immunohistochemistry techniques, and the percentage of MHC II+ cells was measured. The amounts of total CIITA and type I, III and IV CIITA mRNA of lung tissues were measured by real-time PCR using Taqman probe. RESULTS: (1)The percentage of MHC II+ cells in lung tissues increased significantly in fibrosis group compared with that of control group on the 7th day and the 28th day [(0.10 +/-0.03) vs (0.06+/-0.02), P < 0.05; (0.15+/-0.03) vs (0.06+/-0.01),P < 0.01, respectively]; In fibrosis group, the percentage on the 28th day was higher than that on the 7th day [(0.15+/-0.03) vs (0.10+/-0.03), P < 0.05]; (2) Compared with control group on the 7th day, total CIIA mRNA increased 170.4% [(2.89+/-1.07) vs (1.07+/-0.46), P < 0.05], type I CIIA increased 258.8% [(0.77+/-0.38) vs (0.21+/-0.09), P < 0.05], while type IV CIITA decreased 87.2% [(0.39+/-0.15) vs (3.01+/-0.79), P < 0.01]; On the 28th day, total CIITA mRNA increased 98.6% [(4.14+/-1.15) vs (2.08+/-0.76), P < 0.05], type I CIIA increased 137.1% [(0.79+/-0.34) vs (0.33+/-0.23), P < 0.05], type IV CIITA mRNA still decreased, but there was no significant difference [(2.98 +/-0.92) vs (3.95+/-0.93), P > 0.05]; In fibrosis group, type IV CIIA mRNA was 667.3% [(2.98+/-0.92) vs (0.39+/-0.15), P < 0.01] higher on the 28th day than that on the 7th day; Type III CIIA mRNA levels of both groups had no significant difference. CONCLUSION: MHC II/CIITA system of lung tissues was probably involved in the development of rat pulmonary fibrosis.

[Expression of human leukocyte antigen-DR in idiopathic pulmonary fibrosis].

OBJECTIVE: To study the expression of human leukocyte antigen (HLA)-DR in the lungs of the patients with idiopathic pulmonary fibrosis (IPF), and to explore the possible autoimmunity mechanisms of lung fibrosis. METHODS: Methods Immunohistochemistry (SP method) was used to detect the expression of HLA-DR in the lung specimens from 10 IPF patients and in 5 specimens of normal lung tissue immediately adjacent to lung carcinomas as controls. RESULTS: HLA-DR antigens were expressed in the hyperplastic bronchi-alveolar epithelial cells in IPF, but not in the epithelial cells of the normal control lung tissues. The accumulated positive scores of HLA-DR of the IPF group was 27, significantly higher than that of the control group (2, Z = - 3.002, P = 0.001). CONCLUSIONS: Inappropriate HLA-DR expression is present in the bronchi -alveolar epithelium in IPF. Immune dysfunction may play an important role in the development of IPF.

[The effect of angiotensin-(1-7) on the mRNA expression of PDGF and TGF-beta1 in the kidney of diabetic rats].

AIM: To study the effect of angiotensin-(1-7) on the kidney of diabetic rats by observing the mRNA expression of PDGF and TGF-beta1. METHODS: SD rats were divided into three groups: Group C (uni-nephrectomy control group), Group D (diabetic model control group), Group T (Ang-(1-7) treated group). We evaluated blood glucose,urea nitrogen, creatinine and urine albumin excretion respectively, studied the renal morphology by light microscope, and detected the gene expression of PDGF, TGF-beta1 in renal tissue by RT-PCR technique. RESULTS: There was significant difference between the group D and T about the RW/BW, renal morphology, the total urine protein and the mRNA expression of PDGF and TGF-beta1. CONCLUSION: Ang-(1-7) can relieve the renal process of diabetic rats.

Effects of losartan on urinary secretion of extracellular matrix and their modulators in type 2 diabetes mellitus patients with microalbuminuria.

PURPOSE: Angiotensin II receptor Type 1 antagonists postpone the development of nephropathy in type 2 diabetes mellitus (DM). We hypothesize that Losartan may ameliorate renal function in diabetic patients through the regulation on the generation of transforming growth factor (TGF)-beta and fibrinolytic regulators. METHODS: Twenty-two type 2 DM patients with microalbuminuria were treated with 50-100 mg/day of Losartan for 6 months. Urinary secretion of TGF-, plasminogen activator inhibitor-1 (PAI-1), tissue and urokinase plasminogen activators (tPA and uPA) fibronectin, collagen IV and plasma levels of TGF-beta, PAI-1, tPA and uPA of the patients before and after the treatment were analyzed using enzyme-linked immunoabosorbance assay. RESULTS: Losartan effectively reduced arterial blood pressure and urinary albumin excretion. The levels of TGF-beta in urine, but not in plasma, were reduced after 2, 4 and 6 months of the treatment (-32% to -48%, P < 0.05 or 0.01). Urinary or plasma levels of PAI-1, tPA or uPA, and urinary secretion of fibronectin or collagen IV were not significantly altered by Losartan treatment. Urinary levels of collagen IV positively correlated with uPA, and that of fibronectin negatively correlated with PAI-1 in the patients (P < 0.01). Urinary TGF-beta negatively correlated uPA in urine of the patients (P < 0.01). CONCLUSION: Losartan reduced urinary excretion of TGF-beta and albumin in type 2 DM patients with microalbuminuria. Fibrinolytic regulators and TGF-beta are implicated in the regulation of ECM turnover in kidneys of the patients with diabetic nephropathy.