OBJECTIVERotavirus is the single most common cause of severe dehydrating diarrhea in young children worldwide, but the pathogenesis and immunity against this disease are not completely understood. A prospective study was conducted to assess gene expression of toll-like receptors (TLR) in children with acute rotavirus diarrhea.

METHODSSeventy-five children with acute rotavirus diarrhea and 38 control children were enrolled in this study from Sep. 2004 to Jan. 2005. All the 75 patients had detailed records of clinical characteristics. Rotavirus antigen was detected by ELISA from stools. Peripheral blood mononuclear cells (PBMC) were separated by Ficoll reagent and RNA was extracted by Trizol. The levels of mRNA for five TLRs in PBMC were examined by fluorescent quantitative RT-PCR.

RESULTSPatients with acute rotavirus infection had elevated mean levels of TLR 2, 3, 4, 7, 8 mRNA expressions in PBMC within 3 days since onset of the disease, P less than 0.05. But only TLR 2, 3, 8 mRNA levels remained increased in patients within 7 or 14 days since onset (P less than 0.05). Mean levels of mRNA for TLR 4 in PBMC was higher in patients with more severe diarrhea including longer duration of diarrhea, more episodes of diarrhea per day and higher severity scores (P less than 0.05).

CONCLUSIONManifold TLR may play roles in the start-up and regulation of immune responses in children with acute rotavirus diarrhea. These findings will be helpful to further recognize immune response in Chinese children with rotavirus diarrhea and, consequently, may provide directions and insights that could prove critical to the prevention or treatment of this important disease.

Acute Disease ; Antigens, Viral ; analysis ; Child, Preschool ; Diarrhea ; genetics ; virology ; Enzyme-Linked Immunosorbent Assay ; Feces ; virology ; Gene Expression ; Host-Pathogen Interactions ; Humans ; Infant ; Leukocytes, Mononuclear ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus ; immunology ; physiology ; Rotavirus Infections ; genetics ; virology ; Toll-Like Receptors ; genetics

OBJECTIVETo investigate the expression of cathepsin D in ovary of patients with polycystic ovarian syndrome (PCOS).

METHODSWestern blot was performed to detect the expression of cathepsin D and immunohistochemistry was used to detect the protein distribution in ovarian tissue.

RESULTSemi-quantity values of cathepsin D expression in PCOS and control group were 2.06 +/- 0.39 and 4.76 +/- 1.43 (P<0.05), respectively. Immunostaining for cathepsin D was obvious in both follicles and stromal cells, and the strongest immunostaining was seen in granulosa cells of follicles. Immunochemical study showed the protein was mainly located on the cytoplasm and cell membrane.

CONCLUSIONCathepsin D expression is down-regulated in ovaries of PCOS patients, which may provide a clue for the abnormality of follicle development in PCOS.

Adult ; Blotting, Western ; Cathepsin D ; biosynthesis ; Down-Regulation ; Female ; Humans ; Immunohistochemistry ; Ovary ; enzymology ; pathology ; Polycystic Ovary Syndrome ; enzymology

OBJECTIVE: To evaluate the immunological function in rats with formalin inflammatory pain through intrathecal pumping different dosages of morphine. METHODS: Thirty-two Sprague-Dawley rats were randomly divided into 4 groups (n = 8 in each group): saline group (NS) and morphine group included M1 group (10 microg/h) , M2 group (5 microg/h), and M3 group (2.5 microg/h). Chronic intrathecal catheterization was performed under anesthesia with 10% chloral hydrate (300-350) mg/kg according to M2 group (5 microg/h) and M3 group (2. 5 microg/h). Chronic intrathecal catheterization was modified Yaksh's. After 7 days, pain intensity scoring (PIS) was utilized to assess antinociceptive effect of morphine. And spleens were aseptically removed to obtain splenic cells. T lymphocyte function was evaluated based on Concanavalin-A induced splenocyte proliferation. A modified lactic acid dehydrogenase release assay was used to assess NK cell activity. Phenotypic expression of cell surface markers of T lymphocyte subsets (CD3+, CD3+ CD4+, CD3+ CD8+, and CD4+ / CD8+ ) and NK cell ( CD161+) in the spleen were analyzed by flow cytometry. RESULTS: Compared with the NS group, PIS of morphine group decreased obviously (P < 0.01) and was dose-dependent in the early and late phase of formalin pain, but there were no significant differences among morphine groups. Spleen index, splenocyte proliferation and NK cell activity were significantly suppressed by intrathecal pumping morphine. Phenotypic expression of T lymphocyte subsets and NK cell assessed by flow cytometry were different from the control group in all morphine groups. CONCLUSION There was significant antinociception of intrathecal pumping morphine. After intrathecal pumping different dosages of morphine (10 microg/h,5 microg/h, and 2.5 microg/h), the function of cellular immunity was suppressed and was dose-dependent.
Analgesics, Opioid ; administration & dosage ; pharmacology ; Animals ; Dose-Response Relationship, Drug ; Formaldehyde ; Immunosuppressive Agents ; administration & dosage ; pharmacology ; Injections, Spinal ; Killer Cells, Natural ; immunology ; Male ; Morphine ; administration & dosage ; pharmacology ; Pain ; chemically induced ; immunology ; Pain Measurement ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; T-Lymphocyte Subsets ; immunology

OBJECTIVE: To evaluate the effect of intrathecal pumping tramadol on cell-mediated immunity in rats with formalin inflammatory pain. METHODS: Thirty-two Sprague-Dawley adult male rats weighting 250 approximately 300 g were randomly divided into 4 groups (n=8 in each group):Saline group (NS) and 3 tramadol groups (T1,T2,and T3). The rats were anesthetized with intraperitoneal chloral hydrate (300 approximately 350)mg/kg. Microspinal catheter was inserted into the subarachnoid space at the lumber region according to modified Yaksh techniques. In the tramadol groups,after 5 days tramadol was continuously infused through the spinal catheter at 50 (T1),25 (T2), and 12.5 microg/h (T3) for 7 days. In the NS group normal saline was continuously infused instead of tramadol. On Day 7 formalin (5%, 50 microL) was injected into the plantar surface of the left hindpaw. The number of flinches, lickings and total time of licking was recorded for 60 min.Pain intensity scoring(PIS)(0 approximately 3;0= no pain, 3=severe pain) was used to assess the antinociceptive effect of intrathecal tramadol. The rats were killed after the evaluation of pain intensity. Body weight and spleen weight were measured and spleen index (spleen weight/body weight) was calculated. T-lymphocyte function was evaluated based on Concanavalin-A(ConA) induced splenocyte proliferation. A modified lactic acid dehydrogenase(LDH) release assay was done to assess the NK cell activity. Phenotypic expressions of cell surface markers of T lymphocyte subsets (CD3+, CD3+ CD4+, CD3+ CD8+, and CD4+/ CD8+) and NK cell(CD161+) in the spleen were analyzed by flow cytometry. RESULTS: The PIS scores were significantly lower in the T1,T2,and T3 groups than those in the NS group. The spleen index and splenocyte proliferation induced by ConA were significantly suppressed in the T1 group,and the phenotypes of T lymphocyte subsets were significantly changed,but no significant difference was found in the T2 and T3 groups compared with the NS group. There were no differences in NK cell activity in the 3 tramadol groups from the control group. CONCLUSION Intrathecal pumping tramadol has significantly antinociceptive effect. Intrathecal pumping higher dosage tramadol (50microg/h) suppresses T lymphocyte proliferation and alteres T lymphocyte subset phenotype but does not affect NK cell activity. General analgesic dosage tramadol (25 and 12.5 microg/h) has no effect on the immune function.
Analgesics, Opioid ; pharmacology ; Animals ; Dose-Response Relationship, Drug ; Formaldehyde ; Injections, Spinal ; Killer Cells, Natural ; immunology ; Male ; Pain ; chemically induced ; immunology ; Pain Measurement ; drug effects ; Rats ; Rats, Sprague-Dawley ; T-Lymphocyte Subsets ; immunology ; Tramadol ; administration & dosage ; pharmacology

OBJECTIVETo screen molecular markers in early breast cancer and establish gene subtyping-based diagnostic criteria for predicting the prognosis of early breast cancers.

METHODSTumor tissue specimens were obtained from 8 patients with early breast cancer for analysis of the differentially expressed genes using Agilent custom 8×15 000 chips in combination with the prognostic data of the patients. Another 42 tumor tissue specimens were used to validate the differential genes by real-time fluorescent quantitative PCR.

RESULTSGene microarray analysis identified 132 differentially expressed genes between the patients with favorable and poor prognosis, and 44 of these genes were significantly up-regulated (by over two folds) and 88 down-regulated in patients with poor prognoses.

CONCLUSIONThe gene expression profiles differ in early breast cancer tissues of the same pathological type but with different clinical stages and prognoses, and CD44, MKI67, NTRK2, Nek2, C16orf60, TOP2A, ANCCA, and RRM2 genes can be used as the prognostic markers for early breast cancer.

Adult ; Aged ; Biomarkers, Tumor ; analysis ; genetics ; Breast Neoplasms ; genetics ; pathology ; Carcinoma, Ductal, Breast ; genetics ; pathology ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Middle Aged ; Neoplasm Staging ; Oligonucleotide Array Sequence Analysis ; Prognosis

OBJECTIVETo investigate the effects of non-surgical periodontal therapy on serum inflammatory factors and metabolism levels in obese rats with experimental periodontitis.

METHODSSixteen obese rats with experimental periodontitis were randomly divided into treatment group and control group with non-surgical periodontal therapy and no treatment, respectively. Oral glucose tolerance test was performed before treatment and 2 weeks after the treatment. All the rats were sacrificed 2 weeks after treatment and the orbital vein blood was taken to detect fasting blood glucose, fasting insulin, and serum level of C-reactive protein (CRP). Results Two weeks after periodontal treatment, fasting blood glucose (t=2.445, P=0.034) and beta cell function index (t=-2.543, P=0.027) were significantly lower in the treatment group than in the control group. Compared with those in the control group, CRP level (t=2.388, P=0.028) and the area under the curve in the oral glucose tolerance test (t=12.053, P=0.000) decreased significantly in the treatment group.

CONCLUSIONNon-surgical periodontal treatment can reduce serum CRP level and improve glucose metabolism in obese rats.

OBJECTIVETo investigate the role of epidermal growth factor receptor (EGFR) signaling pathway in bronchial epithelial actin stress fiber (F-actin) rearrangement induced by house dust mite (HDM).

METHODSNormal human bronchial epithelial cells (16HBE) were stimulated with HDM with or without pretreatment with AG-1478, an EGFR inhibitor. The levels of phospho(p)-EGFR, F-actin, E-cadherin and β-catenin in the cell cultures were detected with Western blotting. The localizations of F-actin, E-cadherin and β-catenin in the bronchial epithelial cells were determined with immunofluorescence assay, and the transmembrane electrical resistance (TER) and FITC-dextran flux (FITC-DX) in the cells were measured to assess the barrier function of the bronchial epithelia.

RESULTSHDM stimulation of the cells for 10 min resulted in significantly increased p-EGFR expression (P<0.05) without causing obvious changes in the expression of E-cadherin (P>0.05) or β-catenin (P>0.05). Immunofluorescence assay revealed delocalization of E-cadherin and β-catenin in HDM-treated 16HBE cells, shown by their diffusion from the cell membrane to the cytoplasm. In HDM-treated cells, the TER was significantly decreased to (70.00∓4.33)% and the FITC-DX was significantly increased to (115.98∓4.34)%; Inhibition of EGFR reversed the delocalization of E-cadherin and β-catenin, improved the TER to (90.00∓3.75)% and lowered the FITC-DX to (101.10∓2.10)%. HDM induced increased expression and rearrangement of F-actin, which was obviously inhibited by pretreatment of the cells with AG-1478 (P<0.05).

CONCLUSIONEGFR signaling pathway mediates HDM-induced F-actin rearrangement in human bronchial epithelial cells to contribute to epithelial barrier dysfunction.

This study investigated the relationship between IL-33/ST2 signal pathway gene polymorphisms and myocardial infarction (MI) in Han Chinese. A case-control association analysis was performed on a total of 490 MI patients (MI group) and 929 normal subjects (NC group). Sequenom Mass Array and Taqman genotyping technique were used to analyze the tag single nucleotide polymorphisms (SNPs) in the genes encoding IL-33, ST2, and IL-1RaP (rs11792633, rs1041973 and rs4624606). The results showed that the frequencies of rs4624606 genotypes AA, TT, AT were 0.031, 0.647, 0.322 in MI group and 0.026, 0.712, 0.263 in NC group, and the allele frequencies of A and T were 0.192, 0.808 in MI group and 0.157, 0.843 in NC group. There were significant differences in rs4624606 genotypes and allele frequencies between MI group and NC group (P<0.05). For rs11792633, the allele frequencies of C and T were 0.45, 0.55 in MI group and 0.454, 0.546 in NC group with no significant differences found between the two groups. Compared with genotype CC+TC, rs11792633 genotype TT had an increased risk of hypertension (P<0.05). However, there were no significant differences in the frequencies of rs11792633 genotypes between the two groups. No significant differences were noted in the frequencies of rs1041973 genotype and allele between the two groups. Logistic regression analysis showed that rs4624606 genotypes AT and AA+AT were both significantly associated with MI (AT: OR=1.325, P=0.029, 95% CI=1.03-1.705; AA+AT: OR=1.316, P=0.028, 95% CI=1.03-1.681) after factors such as age, gender, smoking, drinking, body mass index (BMI), triglyceride (TG) and cholesterol were adjusted. Those carrying rs4624606 genotype AT or AA+AT had an increased risk of MI. No associations were found between the polymorphisms of the other two loci with MI. It was concluded that, in the IL33/ST2 signal pathway, the A allele of rs4624606 polymorphism of IL-1RaP gene is a potential independent risk factor for MI, and the genotypes AA+AT and AT are associated with the incidence of MI.
China ; Ethnic Groups ; genetics ; Female ; Humans ; Interleukin-1 Receptor-Like 1 Protein ; Interleukin-33 ; Interleukins ; genetics ; metabolism ; Male ; Myocardial Infarction ; genetics ; Receptors, Cell Surface ; genetics ; metabolism ; Signal Transduction ; genetics

OBJECTIVETo study the clinicopathologic features and differential diagnosis of cutaneous regressing/regressed melanoma.

METHODSHistopathologic evaluation and immunohistochemical study by EnVision method were performed in 8 cases of cutaneous regressing/regressed melanoma. The clinical presentation, treatment and follow-up data were analyzed.

RESULTSThe age of the patients ranged from 40 to 69 years (mean 58 years). The male-to-female ratio was 3: 1. Tumors were located on the back (4 cases), sole of the foot (2 cases), ventral aspect of the toes (1 case), and the forearm (1 case). Clinically, 6 patients presented with progressive black mole of the skin, followed by subsequent focal hypopigmentation, even scarring. Two patients presented with multiple foci of dark-brown pigmentation. Microscopically, 3 cases were completely regressed malignant melanoma. Tumoral melanosis was found in 1 of 3 cases. The other 5 cases were melanoma with severe regression. The extent of regression ranged from 75% to 90%. The Breslow depth of the tumors ranged from 0.5 to 1.0 mm. Immunohistochemically, both metastatic and primary tumor cells were diffusely positive for S-100, HMB45 and Melan A, while melanophages were positive for CD68. Follow-up data were available in 8 patients, ranging from 8 to 27 months. Five patients were alive with no evidence of disease, 1 patient was alive with stable disease and 2 patients died of metastatic melanoma.

CONCLUSIONSCorrelation between clinical presentation and pathologic features is important for diagnosis of cutaneous regressing/regressed melanoma. Thin melanoma with extensive regression ( ≥ 75%) should not been regarded as low metastatic risk and wide excision combined with sentinel lymph node biopsy is recommended.

Adult ; Aged ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Back ; pathology ; Female ; Follow-Up Studies ; Foot ; pathology ; Humans ; Lymphatic Metastasis ; MART-1 Antigen ; metabolism ; Male ; Melanoma ; metabolism ; pathology ; surgery ; Melanoma-Specific Antigens ; metabolism ; Melanosis ; pathology ; Middle Aged ; S100 Proteins ; metabolism ; Sentinel Lymph Node Biopsy ; Skin Neoplasms ; metabolism ; pathology ; surgery ; Toes ; pathology ; Treatment Outcome

OBJECTIVETo investigate the clinical and computed tomography (CT) appearances of pulmonary mucosa-associated lymphoid tissue (MALT) lymphoma.

METHODSThe CT findings and clinical data of 13 patients with pathologically proven pulmonary MALT lymphoma were retrospectively reviewed.

RESULTSAmong these 13 patients, seven presented no notable abnormalities, six manifested respiratory symptoms including cough, expectoration, and dyspnea; one of these six patients experienced fever. Chest CT showed solitary nodule in 2 patients and multiple nodules in 3 patients; meanwhile, it showed solitary consolidation in 3 patients and multiple consolidations in 5 patients. Other CT findings included air bronchogram (n = 13), airway dilatation (n = 4), ground glass opacities (n = 5), and interstitial changes (n = 5). One patient had mediastinal lymphoadenopathy and 2 had pleural effusion. Pathology showed massive lymphocyte infiltration; cells with notable nuclear atypia were also seen, which were generated from B cells.

CONCLUSIONSThe main CT findings of pulmonary MALT lymphoma include nodules, mass or patchy consolidations with air brochogram; hilar and mediastinal lymphadenopathies are rare. Clinical diagnosis should also be based on pathological findings and immunohistochemical results.

Adult ; Aged ; Female ; Humans ; Lung Neoplasms ; diagnosis ; diagnostic imaging ; pathology ; Lymphoma, B-Cell, Marginal Zone ; diagnosis ; diagnostic imaging ; pathology ; Male ; Middle Aged ; Radiography ; Retrospective Studies