Effect of antivirals plus low-dose, short-term glucocorticoids on post-herpetic neuralgia.

BACKGROUND: Post-herpetic neuralgia (PHN) is the most common complication of herpes zoster (HZ). It is unclear whether short-term and low-dose glucocorticoids combined with antivirals can reduce the incidence of PHN. OBJECTIVES: To investigate the effects of antivirals plus low-dose, short-term glucocorticoids on PHN. MATERIALS & METHODS: A total of 394 patients with HZ were divided into glucocorticoid and non-glucocorticoid groups, and the incidence of PHN was studied retrospectively. Forty patients with HZ were randomized into the glucocorticoid (n=20) and non-glucocorticoid (n=20) groups. The levels of protein 100-B (S100B) and neuron-specific enolase (NSE) in the blood and the viral load in the skin lesions were measured before and after seven days of treatment. Patient condition and pain were assessed using the HZ and visual analogue scale pain scores. RESULTS: The incidence of PHN in the glucocorticoid and non-glucocorticoid groups was 20.89% and 30.51%, respectively. In patients with onset time >seven days before treatment, the incidence of PHN was 19.81% and 40.16%, respectively. In the glucocorticoid group, after treatment, the mean serum NSE level of the glucocorticoid group decreased from 15.8 ng/mL to 14.0 ng/mL, while the mean serum S100B level decreased from 1486.3 ng/mL to 1453.7 ng/mL. There was no intergroup difference in the reduction rate of viral load. The mean condition score and pain score were significantly lower in the glucocorticoid group. CONCLUSION: Antiviral therapy plus low-dose, short-term glucocorticoids can improve the condition of patients with HZ and partly reduce the incidence of PHN.

Chlamydial-Secreted Protease Chlamydia High Temperature Requirement Protein A (cHtrA) Degrades Human Cathelicidin LL-37 and Suppresses Its Anti-Chlamydial Activity.

BACKGROUND Chlamydia trachomatis is an obligate intracellular pathogen that can cause severe reproductive tract complications while ascending infection occurs. When spreading from cell to cell in a host, C. trachomatis utilizes various survival strategies to offset host defense mechanisms. One such strategy is to degrade host antimicrobial defense proteins before they can attack the invading C. trachomatis cells. MATERIAL AND METHODS We expressed and purified recombinant chlamydia high temperature requirement protein A (cHtrA) including 2 cHtrA mutants (MT-H143A and MT-S247A), and also extracted endogenous cHtrA. Proteins were identified and their purity evaluated by SDS-PAGE and Western blot. The anti-chlamydial activity and degradation of 5 antimicrobial peptides (cathelicidin LL-37, alpha-defensin-1 and -3, and ß-defensin-2 and -4) by cHtrA and 2 cHtrA mutants (MT-H143A and MT-S247A) were tested by immunoassay and Western blot. RESULTS Of the 5 antimicrobial peptides (cathelicidin LL-37, alpha-defensin-1 and -3, and ß-defensin-2 and -4) tested, cathelicidin LL-37 showed the strongest anti-chlamydial activity. Interestingly, cHtrA effectively and specifically degraded LL-37, suppressing its anti-chlamydial activity. The 2 cHtrA mutants (MT-H143A and MT-S247A) were unable to degrade LL-37. Comparison of cHtrA activity from C. trachomatis D, L2, and MoPn strains on LL-37 showed similar responses. CONCLUSIONS cHtrA may contribute to C. trachomatis pathogenicity by clearing the passage of invasion by specific LL-37 degradation.

Chlamydia muridarum plasmid induces mouse oviduct pathology by promoting chlamydial survival and ascending infection and triggering host inflammation.

BACKGROUND: Infection with plasmid-free Chlamydia trachomatis and Chlamydia muridarum fails to induce severe pathology, however, the mechanisms underlying this phenotype are unclear. OBJECTIVES: To elucidate the mechanisms of chlamydial plasmid-mediated pathology in mouse oviducts. MATERIALS & METHODS: BALB/c mice were intravaginally infected with either plasmid-competent or plasmid-free C. muridarum strains. To compare the survival and ascending infection of these strains, vaginal swabs and genital tract tissues were collected and cultured with HeLa cells to monitor the recovery of live organisms. In addition, Chlamydia strains were intrabursally inoculated into the oviducts of mice to assess pathogenicity. Cytokine levels in the vaginal swabs collected from both the plasmid-competent and plasmid-free C. muridarum-infected mice were detected using Bio-Plex Pro Mouse Cytokine, Chemokine, and Growth Factor Assays. RESULTS: The plasmid-competent C. muridarum strain induced hydrosalpinx formation in mouse oviducts following intravaginal inoculation, however, this was not the case for the plasmid-free C. muridarum strain. The lack of hydrosalpinges in response to the plasmid-free C. muridarum strain correlated with its significantly reduced ability to survive and disseminate to the upper genital tract. Furthermore, the plasmid-free C. muridarum failed to induce hydrosalpinx formation in mice, even when the strain was intrabursally injected into oviducts. A comparison of the cytokine levels in mouse vaginal secretions showed that the plasmid-free C. muridarum strain induced less IL-15, LIF, MIP-2, IL-1α, IL-1ß, TNF-α, and RANTES. CONCLUSION: C. muridarum plasmid contributes to oviduct pathology by promoting bacterial survival and ascending infection, and triggering host inflammatory responses.

Chlamydial Plasmid-Encoded Protein pGP3 Inhibits Development of Psoriasis-Like Lesions in Mice.

BACKGROUND The anti-microbial protein cathelicidin LL-37 plays an important role in the pathogenesis of psoriasis by inducing inflammation. Our previous study showed that the chlamydial plasmid-encoded protein pGP3 forms a stable complex with LL-37 to neutralize its pro-inflammatory activity. Here, we explored whether pGP3 can inhibit the development of lesions in mice with imiquimod-induced psoriasis. MATERIAL AND METHODS The protein pGP3 was expressed in bacteria and purified using glutathione-conjugated agarose beads and a precision protease. The ability of the purified pGP3 to block chemotaxis mediated by LL-37 was tested in vitro using bone marrow-derived neutrophils. The ability of the protein to inhibit the development of psoriasis-like lesions was tested by topically or subcutaneously administering pGP3 in doses of 10 or 50 µg to mice previously treated with imiquimod. Mouse skin was evaluated using the psoriasis area and severity index (PASI) score and photography. Skin biopsies were taken on day 8 and analyzed histologically. RESULTS Purified pGP3 inhibited LL-37-mediated chemotaxis. Mice treated with 50 µg pGP3 showed clinical improvement with less severe erythema, infiltration, and scales; these mice also showed thinner dermis and less hyperkeratosis, parakeratosis, and inflammatory cell infiltration than mice treated with without 10 µg pGP3. CONCLUSIONS PGP3 can inhibit the development of psoriasis-like lesions in mice, possibly through its ability to bind LL-37. Future work should examine the mechanisms underlying this therapeutic effect.

IL-37b gene transfer enhances the therapeutic efficacy of mesenchumal stromal cells in DSS-induced colitis mice.

AIM: To investigate whether the transfer of the IL-37b gene, a newly identified inhibitor of both innate and adaptive immunity, could improve the therapeutic efficacy of mesenchumal stromal cells (MSCs) in inflammatory bowel disease (IBD). METHODS: The expression of IL-37 in biopsied specimens of the patients with active ulcerative colitis (UC) was detected using RT-PCR and immunohistochemistry. Mice were treated with 3% dextran sulfate sodium (DSS) for 8 days to induce colitis. Before DSS treatment, the mice were injected with MSCs, MSC-eGFP or MSC-IL37b. Their body weight was measured each day, and the colons and spleens were harvested on d 10 for pathological and biochemical analyses. RESULTS: In biopsied specimens of the patients with active UC, the expression of IL-37 was dramatically elevated in inflamed mucosa, mainly in epithelial cells and infiltrating immune cells. Compared to MSC-eGFP or MSCs, MSC-IL37b administration significantly attenuated the body weight and colon length reduction, and decreased the histological score in DSS-induced colitis mice. Furthermore, MSC-IL37b administration increased the percentage of myeloid-derived suppressor cells (MDSCs) among total splenic mononuclear cells as well as the percentage of regulatory T cells (Tregs) among splenic CD4+ T cells in the mice. Moreover, MSC-IL37b administration increased the IL-2+ cells and decreased the IFN-γ+ cells among splenic CD4+ T cells. CONCLUSION: IL-37 is involved in the pathophysiology of UC. IL-37b gene transfer enhances the therapeutic efficacy of MSCs in DSS-induced colitis mice by inducing Tregs and MDSCs and regulating cytokine production.

[Detection of EGFR and COX-2 expression by immunohistochemical method on a tissue microarray section in lung cancer and biological significance].

BACKGROUND AND OBJECTIVE: Epidermal growth factor receptor (EGFR) and cyclooxygenase-2 (COX-2), which can regulate growth, invasion and metastasis of tumor through relevant signaling pathway, have been detected in a variety of solid tumors. The aim of this study is to investigate the biological significance of EGFR and COX-2 expression in lung cancer and the relationship between them. METHODS: The expression of EGFR and COX-2 was detected in 89 primary lung cancer tissues, 12 premalignant lesions, 12 lymph node metastases, and 10 normal lung tissues as the control by immunohistochemical method on a tissue microarray section. RESULTS: EGFR protein was detectable in 59.6%, 41.7%, and 66.7% of primary lung cancer tissues, premalignant lesions and lymph node metastases, respectively; COX-2 protein was detectable in 52.8%, 41.7%, and 66.7% of primary lung cancer tissues, premalignant lesions and lymph node metastases, respectively, which were significantly higher than those of the control (P < 0.05). The positive ratios and the levels of the expression of EGFR and COX-2 proteins were closely related to histological type, clinical stage and lymph node metastasis of lung cancer (P < 0.05), but not to histological grade, sex and age (P > 0.05). COX-2 expression was related to gross type (P < 0.05). A highly positive correlation was observed between EGFR and COX-2 expression (P < 0.01). CONCLUSION: Overexpression of EGFR and COX-2 may play an important role in the tumorgenesis, progression and malignancy of lung cancer. Detection of EGFR and COX-2 expression might be helpful to diagnosis and prognosis of lung cancer.

[Development and clinical application of frozen tissue microarray in lung cancer diagnosis.].

BACKGROUND: Traditional techniques in clinical diagnostic pathology have some limitations. Here we tried to develope frozen tissue microarray as a fast, simple and economical technique. METHODS: A total of 48 dots frozen tissue microarray were constructed in this study including primary 19 lung cancer samples and 5 normal lung tissue samples from primary lung cancer. Hematoxylin-Eosin and immunohistochemical stains were performed on the sections and we assess the feasibility on morphology and genetic expression. RESULTS: The cores on frozen tissue microarray of OCT block were regularly arranged and had no shift and distort. After HE-stained, morphology of the cores was well preserved. They were answered for the request of research.Immunohistochemistry of array slides: we performed immunohistochemistry on the tumor tissue microarray with antibodies for EMA. EMA staining is uniform across the sample and gives the expected cytochylema-associated staining. There is no background staining. We compared the staining of immunohistochemistry on section of frozen tissue microarray to the results on normal slides. The results showed the accord rates were 94.7% (18/19), and there were no significant differences between them each other (P >0.05). CONCLUSIONS: The frozen tissue microarray is feasible. HE and immunohistochemical stains showing this methodology will be useful for morphology and immunohistochemical based protein analyses.

[Clinicopathological analysis of biliary tract complications on post-liver transplantation patients].

OBJECTIVE: To analyze the pathohistological changes of the livers and the clinical features of patients with biliary tract complications after their orthotopic liver transplantations. METHODS: From Sept 1998 to June 2005 clinical and pathological data of 173 post-liver transplantation patients with biliary tract complications were analyzed. RESULTS: Biliary tract complications occurred within 3-2920 days after the transplantation operations. These complications occurred within 1-30 days, 31-90 days, 91-180 days, 180 days at rates of 49.71%, 17.92%, 4.62%, 27.74% respectively. The complications were of inflammatory nature in 171 cases, (72.25%), and of obstructive nature in 164 cases (27.74%). The main pathological changes were epithelium degeneration of interlobular bile ducts, inflammatory cell infiltration in portal areas, proliferation of interlobular bile ducts, fibrosis in portal areas, cholestasis in small bile ducts and hepatocytes. CONCLUSION: Many of the biliary tract complications of post-liver transplantation in our cases were of inflammatory nature and they often occurred within 30 days after the surgery. Obstructive nature complications often occurred in 90 days after the surgery and the prognosis of these cases was much poorer. The pathological changes of live tissues shown in liver biopsies are important for prognostic evaluation, differential diagnosis and categorization of biliary tract complications.

Expression and significance of Survivin mRNA in lung cancer tissue microarray detected by FISH.

OBJECTIVE: To investigate the expression of Survivin mRNA in lung cancer tissue microarray (TMA) by fluorescence in situ hybridization (FISH) method, and determine the role and significance of it in lung cancer genesis and progress. METHODS: The expression of Survivin mRNA was detected by FISH method and TMA technology. Fifty-four cases of lung cancer and 10 cases of normal lung tissue were examined. RESULTS: Survivin mRNA was expressed in 66.7% (36/54) of lung cancer; the positive ratio of lung cancer was significantly higher than that of normal lung tissue (0/10; chi2 = 15.238, P < 0.05). The positive ratio of Survivin mRNA was significantly higher in poor differentiated cancer (20/24, 83.3%) than moderate and well differentiated cancer (16/30, 53.3%; chi2 = 5.40, P < 0.05). The positive ratio of Survivin mRNA was significantly higher in group with lymph node metastasis (27/32, 84.4%) than without lymph node metastasis (9/22, 40.9%; chi2 = 11.084, P < 0.05). The positive ratio of Survivin mRNA was significantly higher in stage III-IV(12/13, 92.3%) than stage I - II (24/41, 58.5%; chi2 = 5.066, P < 0.05). CONCLUSION: Survivin mRNA highly expresses in lung cancer, which is related to the progress and malignant behavior. Survivin may play a promoting role in lung cancer genesis and progress and provide a basis for estimating prognosis and treatment.

[Expression of KAI1, MRP-1, and FAK proteins in lung cancer detected by high-density tissue microarray].

BACKGROUND & OBJECTIVE: The genesis, development, invasion, and metastasis of tumor are closely correlate with alterations of multi-genes. At present, the roles of Kang-ai-1 (KAI1), motility-related protein-1 (MRP-1), and focal adhesion kinase (FAK) in lung cancer have seldom been reported. This study was designed to investigate the roles of KAI1, MRP-1, and FAK in tumorigenesis and development of lung cancer, and their values in diagnosis and predicting the prognosis of lung cancer. METHODS: The expression of KAI1, MRP-1, and FAK proteins in a high-density tissue microarray containing 240 spots were detected by SP immunohistochemistry. RESULTS: The positive rates of KAI1 and MRP-1 were significantly lower in primary lung cancer than in normal lung tissue (25.9% vs. 100%, 42.6% vs. 100%, P<0.05). The positive rate of FAK was significantly higher in primary lung cancer than in normal lung tissue (44.4% vs. 10.0%, P<0.05). The expression of KAI1, FAK, and MRP-1 in primary lung cancer had no correlation with age and gender of the patients, and macroscopic and histological type of tumor, but had correlations with tumor differentiation, clinical stage, and lymph node metastasis. In addition, the expression of MRP-1 had correlation with histological type of tumor; the positive rate of MRP-1 was significantly lower in small cell lung cancer than in non-small cell lung cancer (0 vs. 50.0%, P<0.05). KAI1 expression was negatively correlated to FAK expression (rs=-0.458, P<0.05); MRP-1 expression was positively correlated with KAI1 expression (rs=0.535, P<0.05), and negatively correlated with FAK expression (rs=-0.438, P<0.05). CONCLUSIONS: The abnormal expression of KAI1, MRP-1, and FAK proteins are related to invasion and metastasis of lung cancer. Combined detection of the 3 proteins may be useful in predicting the development and prognosis of lung cancer.

[Expressions of EphB4 and HIF-1α in human lung cancer and their significances].

BACKGROUND: It has been proved that EphB4 and HIF-1α are closely related to the oncogenesis and development of lung cancer. The aim of this study is to investigate the biological significances of EphB4 and HIF-1α in lung cancer and their relationship with each other. METHODS: The expression of EphB4 and HIF-1α was detected in 54 lung cancer tissues and 10 normal lung tissues as control by immunohistochemical method. RESULTS: EphB4 and HIF-1α proteins were detectable in 50.0% and 42.6% of all 54 lung cancer tissues respectively, which were significantly higher than those of the control (P < 0.05); the positive ratios and the levels of the expressions of EphB4 and HIF-1α proteins were closely related to gross types, differentiations and clinical stages (P < 0.05), but not to histological classification, age, sex and lymph node metastasis (P > 0.05). A highly positive correlation was observed between EphB4 and HIF-1α expression (P < 0.01 ). CONCLUSIONS: Overexpression of EphB4 and HIF-1α may play an important role in the pathogenesis, progression and malignant degree of lung cancer. Detection of EphB4 and HIF-1α expression might be helpful to predict prognosis of patients with lung cancer.

[Expression of KAI1 protein in tissue microarray and its biological significances in patients with lung cancer].

BACKGROUND: KAI1 is a new identified metastasis-suppressor gene whose expression in many types of tumors has been reported. The aim of study is to investigate the role of KAI1 protein in development of lung cancer and its values in predicting the prognosis of lung cancer. METHODS: The expressions of KAI1 protein were detected in benign pulmonary disease tissue, precancerous disease tissue, lung cancer tissue and metastatic lung cancer tissue in local lymph node using tissue microarray and immunohistochemical method. The relationship between expression of KAI1 protein and clinicopathological parameters of patients with lung cancer was analyzed by Chi-Square test and Fisher exact test. RESULTS: The positive rate of KAI1 expression was 100.0% in 10 cases of benign pulmonary diseases, 66.7% in 12 cases of precancerous diseases, 24.7% in 89 cases of primary lung cancer and 0 in metastatic lung cancer tissue in local lymph node respectively. The KAI1 protein expression in primary lung cancer tissues had no remarkable relationship with age and gender of the patients and the location of cancer, but had significant relationship with the histological type and differentiated degree of tumor, P-TNM stages and lymph node metastatic status. CONCLUSIONS: The abnormal expression of KAI1 protein may participate in malignant progression of lung cancer. Its downregulation may promote the invasion and metastasis of tumor cell. Detection of the expression of KAI1 protein may be helpful to predict the prognosis of lung cancer.

[Expression and significance of matrix metalloproteinases and their tissue inhibitors in the infiltration and metastasis of lung cancer].

BACKGROUND: To determine the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in lung cancer specimens, and to find out the possible roles of MMPs and TIMPs in the infiltration and metastasis of lung cancer. METHODS: The expression of MMP-1, MMP-2, MMP-9, MMP-13, TIMP-1, TIMP-2 were detected in 104 lung cancer tissues by immunohistochemical method. RESULTS: The expressions of MMPs and TIMPs were up-regulated in lung cancer tissues. The expression of MMP-2 was related to differentiated degree of tumor cells. MMP-9 correlated with lymph node metastasis of lung cancer. The positive rate of TIMP-1 was related to TNM stage and lymph node metastasis. In lung cancer tissues, there was positive correlation between MMP-2 and TIMP-2, and between MMP-9 and TIMP-2. CONCLUSIONS: MMPs and TIMPs may play the important roles in the development of lung cancer. MMP-9 and TIMP-1 might promote the infiltration and metastasis of lung cancer.

[Relationship between the expression of E-cadherin and MVD and its significance in lung cancer].

BACKGROUND: To study the relationship between the expression of E-cadherin and MVD in lung cancer and its significance. METHODS: The expressions of E-cadherin and factor VIII were detected in 104 lung cancer tissues by immunohistochemical method, and MVD was calculated by image analysis system. RESULTS: The expression of E-cadherin was significantly related to the differentiation of lung cancer (P < 0.05). A negative correlation was observed between E-cadherin expression and MVD in lung cancer tissues (P= 0.047). CONCLUSIONS: Downexpression of E-cadherin and increase of MVD may play an important role in the invasion and metastasis of lung cancer, and may also be used as a useful marker for tumor prognosis.