Objective To investigate the kinesin family member 17 (KIF17) expression and cellular localization in the hippocampus and temporal lobe cortex in the rat lithium-pilocarpine model of epilepsy,and discuss its function in the epilepsy pathogenesis.Methods The animal model was established by lithiumpilocarpine induction in rats.Totally 49 adult healthy male Wistar rats were randomly divided into control group (n =7) and experimental group (n =42).The experimental group included 6 subgroups (n =7)according to sacrifice time (24 h,72 h,7 d,14 d,1 month and 2 months).The expression and localization of KIF17 were examined by western blot and double-label immunofluorescence,respectively.Results In rat hippocampus,the expression of KIF17 protein increased after the onset of seizure (the ration of KIF17/β-actin were:24 h 0.516 ± 0.196,72 h 0.742 ± 0.313),reached its peak in 7 days (0.888 +0.319)and then slowed down (14 d 0.770 ± 0.271,1 month 0.742 ± 0.261,2 months 0.714 ± 0.271),all of which were significantly higher than that in the control group (0.495 ± 0.203).And all the differences had statistical significance (t =7.051,4.974,7.419,8.795,8.264,6.676,all P ＜ 0.05).In rat cortex of temporal lobe,the expression of KIF17 protein increased after the onset of seizure and reached its peak in 30 d.The optical density ration in the experimental groups were higher than that in the control group.Doublelabel immunofluorescence disclosed that the KIF17 localized in the neurons,including excitable neurons and inhibitory neurons,but not in the astrocytes which were performed with anti-microtubule-associated protein 2,anti-brain-specific Na-dependent inorganic phosphate cotransporter,anti-glutamate decarboxylase 1 and anti-glial fibrillary acidic protein,respectively.Conclusion KIF17 may play a potential role in the pathogenetic mechanisms of the rat lithium-pilocarpine model of epilepsy.

Objective: To evaluate the effect of nutritional support for lung transplantation patients.Methods: The lung transplantation patient received perioperative enteral nutrition(EN) .Exogenous glutamine(Gln) and recombinant human growth hormone(rhGH) were postoperatively used for 7-14 days.Results: The patients weight increased from 53 kg to 55 kg. No respiratory failure and acute rejection occurred postoperatively. The patient recovered fluently.Conclusion: Appropriate perioperative nutritional support and postoperative metabolic intervention can facilitate the recovery of lung transplant patient.

AIDS-Related Opportunistic Infections ; diagnosis ; Humans ; Oral Medicine ; Sarcoma, Kaposi ; diagnosis

Objective To investigate the changes of CD4+ CD25+ regulatory T lymphocyte (Treg) and expressions of folkhead helix transcription factor 3 (FoxP3) in intestinal mucosa in human immunodeficiency virus (HIV) infected patients. Methods Twenty-one HIV infected patients and 17 control subjects without HIV infection were included in this study. The expression of FoxP3, which was considered as a specific marker of CD4+ CD25 + Treg, was detected in intestinal mucosa specimens from HIV infected patients by immunohistochemistry. Meanwhile, the in situ expression of CD4+ T lymphocyte was also determined by immunohistochemistry. The data were analyzed by t test. Results The positive labeling index of CD4+ T lymphocyte in intestinal mucosa was significantly lower in HIV infected patients compared to the controls (11. 56%±4. 44% vs 43. 49% ±8. 90% ,t=-11. 86,P<0. 01). The positive labeling index of FoxP3 in intestinal mucosa was also significantly lower in HIV infected patients compared to the controls (0.46% ± 0.20% vs 1. 18% ± 0. 44% ,t= - 5. 98,P<0.01). Conclusion The depletion of CD4+ CD25+ Treg is accompanied with the depletion of CD4 + T lymphocyte and the reduction of FoxP3 expression in intestinal mucosa of HIV infected patients.

OBJECTIVETo investigate gene diagnosis of occult micrometastasis in the mediastinal lymph node in patients with non-small cell lung carcinoma (NSCLC) and to evaluate its prognostic significance.

METHODSWith mRNA expression of mucoid1 (MUC1) gene examined by RT-PCR, 168 mediastinal lymph nodes taken from 37 pN(0) (negative lymph nodes) NSCLC patients (stage Ia approximately IIb) made up the experiment group. Thrity negative lymph nodes from 14 benign lesions and 30 positive lymph nodes from 15 NSCLC patients served as control. The survival difference between MUC1 mRNA-negative and MUC1 mRNA-positive groups was compared by the chi(2) test.

RESULTSUC1 mRNA was not identified in the negative-control group (specificity = 100%), but it was identified in 26 of 30 positive-control samples (sensitivity = 86.7%). MUC1 mRNA was identified in 16 (9.5%) of the experiment group from 12 patients whose TNM stage was up-regulated to stage IIIa. The 3-year survival rate (58.3%) of MUC1 mRNA positive group patients with occult micrometastasis in mediastinal lymph node was lower than the 88.0% of MUC1 mRNA negative group (P < 0.05).

CONCLUSIONOccult micrometastasis in the mediastinal lymph node in NSCLC patients can be diagnosed by MUC1 mRNA expression through RT-PCR. Poor prognosis in some pN(0) NSCLC patients may be associated with nodal occult micrometastasis.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; diagnosis ; secondary ; Female ; Genetic Markers ; genetics ; Humans ; Lung Neoplasms ; diagnosis ; pathology ; Lymphatic Metastasis ; diagnosis ; Male ; Middle Aged ; Mucin-1 ; analysis ; genetics ; Prognosis ; RNA, Messenger ; analysis

Epithelial membrane protein 3 (EMP3) is a trans-membrane signaling molecule with important roles in the regulation of apoptosis, differentiation and invasion of cancer cells, but the detailed is largely still unknown. We analyzed the mRNA levels and methylation statuses of EMP3 in 63 primary breast carcinomas and assessed their correlations with clinicopathologic variables. The expression of EMP3 mRNA in primary breast carcinomas was significantly higher than the expression of 20 normal breast tissues (p<10(-7)). EMP3 overexpression in breast carcinomas was significantly related to histological grade III (p=3.9X10(-7)), lymph node metastasis (p= 0.003), and strong Her-2 expression (p=3.3X10(-6)). Hypermethylation frequencies of EMP3 were detected in 36.5% of breast carcinomas by methylation-specific polymerase chain reaction. However, no significant correlations were found between methylation status of EMP3 and mRNA expression levels as well as other clinical parameters. In conclusion, EMP3 may be a novel marker of tumor aggressiveness. Overexpression of EMP3 in primary breast carcinoma is not associated with DNA methylation.
Adult ; Breast Neoplasms/*genetics/pathology ; Carcinoma/*genetics/pathology ; DNA Methylation ; Epigenesis, Genetic ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Membrane Glycoproteins/*genetics/metabolism ; Middle Aged ; Neoplasm Staging ; RNA, Messenger/metabolism ; Receptor, erbB-2/genetics/metabolism ; Severity of Illness Index

OBJECTIVETo identify the type of iron deposition and describe its amount, distribution and associated lesions, in order to support an etiologic diagnosis for hemochromatosis.

METHODSHematoxylineosin (HE) stain, reticular fiber stain, Masson's stain and Perl's iron stain were used to assess liver biopsies from 31 patients with hemochromatosis. The Ishak scoring system and Deugnier scoring system were used to assess the histological change in liver and to semi-quantify the excess of hepatic iron. Genetic testing results were received from a portion of the patients and used in analysis.

RESULTSOne patient had hereditary (-HFE) hemochromatosis complicated with Gilbert's syndrome, for which the pattern of iron deposition was similar to that of the four patients with Gilbert's syndrome. Iron accumulation appeared as fine granules predominating at the biliary pole of cells and was distributed throughout the lobule with a decreasing gradient spanning from the periportal to centrolobular areas. Mild chronic inflammation was found to be commonly associated with low stage fibrosis.One patient had HFE hemochromatosis complicated with hepatitis B virus infection, and the pattern of iron deposition resembled that in the eight patients with viral hepatitis, wherein the deposition was mainly in the sinusoidal cells and/or portal macrophages. Histological grading and fibrosis staging differed among patients. The five patients with blood disordered showed iron accumulation mainly in the periportal hepatocytes, but mesenchymal iron deposits were also present. The grade of inflammation, as well as of fibrosis,was mild. The five patients with alcoholic disease and the five patients with drug-induced hepatitis showed hepatic iron deposition in swollen or ballooned hepatocytes. The two patients with excessive iron supply showed iron deposition localized within the parenchymal and mesenchymal cells.

CONCLUSIONEtiologic diagnosis of hemochromatosis relies on both the type of iron deposition and the nature of associated lesions. Liver biopsy is necessary for both diagnosis and prognosis.

Biopsy ; Hemochromatosis ; Humans ; Iron ; Liver

Objective:To preliminarily evaluate the effect of Fam114A1 on the biological function of melanocytes.Methods:A375 human melanoma cells was used to construct stably Fam114A1-overexpressing or -inhibited cell line by lentiviral transfection, namely overexpression group and expression inhibition group respectively, and A375 cells transfected with an empty lentivirus served as control group. Real-time fluorescence-based quantitative PCR was performed to evaluate effect of Fam114A1 on the mRNA expression of melanin synthesis-related genes tyrosinase (TYR) , tyrosinase-related protein 1 (TYRP1) , premelanosome protein (PMEL) , microphthalmia-associated transcription factor (MITF) and dopachrome isomerase (DCT) , Western blot analysis was conducted to determine the protein expression of TYR and MITF, methyl thiazol tetrazolium (MTT) assay, Transwell migration and adhesion assays were conducted to assess the effect of Fam114A1 on cellular proliferative activity, migratory and adhesive ability of A375 cells respectively. Statistical analysis was carried out by using one-way analysis of variance and Dunnett- t test. Results:Fluorescence microscopy showed that lentivirus-based transfection efficiency was about 90% in the 3 groups. Compared with the control group (0.850 ± 0.120) , the protein expression of Fam114A1 significantly increased in the overexpression group (1.507 ± 0.170, t = 5.888, P = 0.001) , but significantly decreased in the expression inhibition group (0.397 ± 0.120, t = 4.065, P = 0.007) , suggesting that the stably Fam114A1-overexpressing or -inhibited A375 cell line was successfully constructed. Real-time fluorescence-based quantitative PCR and Western blot analysis showed that the mRNA and protein expression of TYR and MITF were significantly lower in the expression inhibition group than in the control group (all P < 0.01) , but did not differ between the overexpression group and control group (all P > 0.05) . Compared with the control group, the expression inhibition group showed significantly increased cellular proliferative activity and adhesive ability ( P = 0.009, 0.001, respectively) , but significantly decreased migratory ability ( P = 0.005) , while the overexpression group only showed significantly increased migratory ability ( P = 0.021) . Conclusions:Fam114A1 can affect the proliferative activity, migratory and adhesive abilities of A375 cells, and the expression of melanin synthesis-related proteins TYR and MITF in A375 cells. Fam114A1 may be a functional protein involved in regulating the biological activity of melanocytes.

Objective:To design and synthesize photosensitizers with different substituents and to identify its physicochemical characteritics and photodynamic effect on tumor cells.Methods:Two kinds of BODIPY photosensitizers BPOI and BPCI were synthesized through condensation reaction between aldehyde and reactive hydrogen of pyrrole,followed with electrophilic substitution reaction.Physicochemical properties were characterized by 1H NMR,FT-IR and UV-visible absorption spectra and fluorescence emission spectra.The ability to produce reactive oxygen species was detected by BPDF and DCFH-DA.Photodynamic therapy effect on rat glioma C6 cells in vitro was determined by MTT method.Results:Two kinds of BODIPY photosensitizers BPOI and BPCI were successfully synthesized with different substituents,which were confirmed by 1H NMR,FT-IR.Both materials had low toxicity and could be readily taken up by tumor cells.The ability of synthesized photosensitizers to produce reactive oxygen species was strongly influenced by solvent polarity when the substituent belongs to electron-donating group,while no effect was found when the substituent belongs to electron-withdrawing group.Conclusion:Photosensitizer BPOI with electron-donating substituent produces reactive oxygen species with a slow rate in a highly polar environment,while the ability to produce reactive oxgen is greatly improved in a low polarity environment,which is expected to be used for environmental-selective photodynamic therapy in tumor cells.

Objective:To synthesize 5-fluorouracil-nicotinamide (5-FU-NCT) cocrystal and to investigate its physicochemical and biological properties.Methods:The cocrystal of 5-Fu-NCT was prepared through the cooling technology.PXRD,NMR,FTIR and DSC were used to characterize the structure of 5-FU-NCT cocrystal.Solubility was measured by HPLC method.Drug resistant human liver cancer BEL-7402/5-FU cells were treated with 5-FU-NCT cocrystal,the inhibition effect was tested by MTT and HE staining,and cancer cell migration was determined by scratch test.Results:According to PXRD,NMR,FTIR and DSC results,the cocrystal of 5-Fu-NCT had been synthesized successfully.The characteristic diffraction peaks (2θ/°) of the cocrystal were 16.4,20.4,22.3,27.9 and 30.1.The solubility of 5-FU-NCT was 13.5 g/L as measured by HPLC.The antitumor activity tests showed that 5-FU-NCT cocrystal enhanced anticancer effect of 5-FU,and the IC50 of 5-FU and 5-FU-NCT was 129.6 μg/mL and 42.6 μg/mL,respectively.Conclusion:5-Fu-NCT cocrystal have been synthesized successfully through the cooling technology and it shows a more enhanced anticancer effect in comparison to 5-FU on BEL-7402/5-FU cells.