Objective To investigate the expression of peroxisome proliferators-activated receptor ? (PPAR?) in trophoblast and relation between PPAR? ligands and trophoblast invasion.Methods We examined the expression of PPAR? by immunohistochemistry,immunocytochemistry and real time quantitative PCR.We next examined,using the cytotrophoblast culture model,the biological role of PPAR? ligands in vitro.Results PPAR? was mainly localized in the nuclei of villous cytotrophoblast and extravillous cytotrophoblast of cell islands and cell columns.In villous tissue and cultured trophoblast from early first trimester,the level of expression of PPAR? mRNA and protein was 36.0?5.1,13.4?3.1 and 1.35?0.08,1.13?0.11;from late first trimester it was 23.3?5.5,6.1?1.3 and 1.17?0.03,0.86 ?0.05,and the expression of PPAR? was obviously decreased (P

BACKGROUNDEnhanced apoptosis of cytotrophoblasts in early pregnancy is associated with high risk of intrauterine growth retardation and preeclampsia, which are two common pregnant complications. Its etiological factors remain unclear. Cytotrophoblasts share some traits with innate immune cells and may show response to lipopolysaccharide. This study was conducted to demonstrate whether lipopolysaccharide has apoptosis-inducing effects on cytotrophoblast and the role of innate immune reaction in this process.

METHODSCytotrophoblasts were isolated from early pregnant villous tissues and cultured with serum-free medium. Subsequently, cytotrophoblasts were treated with lipopolysaccharide at the concentrations of 0 (control), 25, 50, 100 and 200 ng/ml for 24 hours. Apoptosis of cytotrophoblasts was determined by light microscopy, Hoechst 33258 DNA staining with a fluorescent microscope, transmission electron microscope and annexin V-fluorescein isothiocyanate-conjugated/propidium iodide (PI) staining with flow cytometry. Then expression of caspase-3 was detected by Western blot. Confocal immunofluorescence technique was used to detect tumor necrosis factor alpha expression in cytotrophoblasts. The levels of tumor necrosis factor alpha in the culture medium were detected by enzyme-linked immunosorbent assay.

RESULTSUnder light, fluorescence microscope and transmission electron microscope, characteristic alternations of apoptosis in cytotrophoblasts were observed after lipopolysaccharide treatment. Flow cytometry results showed that lipopolysaccharide significantly increased apoptosis indexes of cytotrophoblasts. Significant statistical differences were found in the above groups (P = 0.01). The mean relative densities of bands corresponding to caspase-3 were significantly increased in groups treated with lipopolysaccharide, as compared with the normal control (P < 0.001). Tumor necrosis factor a expression was found to increase in cytotrophoblasts by confocal immunofluorescence technique and in culture medium by enzyme-linked immunosorbent assay after lipopolysaccharide treatment. A positive correlation was found between tumor necrosis factor a expression and apoptosis indexes of cytotrophoblasts (r = 0.747, P < 0.001).

CONCLUSIONApoptosis of cytotrophoblasts could be induced by lipopolysaccharide, in which innate immune reaction is the important mechanism.

Apoptosis ; drug effects ; Blotting, Western ; Cells, Cultured ; Dose-Response Relationship, Drug ; Flow Cytometry ; Humans ; Immunity, Innate ; drug effects ; Lipopolysaccharides ; pharmacology ; Microscopy, Confocal ; Trophoblasts ; cytology ; drug effects ; Tumor Necrosis Factor-alpha ; analysis ; physiology

The study was aimed to explore the role of gene JWA, a novel retinoic acid responsible and cytoskeleton associate gene, in regulating committed differentiation of HL-60 cell and the molecular mechanism in the course of differentiation and apoptosis of leukemic cells. By using FCM, the changes of CD13, CD14, CD15, CD11b and cell cycles were detected in HL-60 cells treated with ATRA (10(-6) mol/L), Ara-C (10 ng/ml) and TPA (10(-8) mol/L) respectively. The samples were determined by semi-quantitative reverse transcript-polymerase chain reaction (RT-PCR) and Western blot for the expression of JWA, Bcl-2, HSP27 and HSP70 at day 0, 2, 4, 6, 8. The results showed that HL-60 cells committedly differentiated into granulocyte-, monocyte-, macrophage-like cells. As a result, JWA was up-regulated in a time-dependent manner, while Bcl-2 was down- regulated at the same time. In ATRA and TPA group, the change of HSP70 had positive correlation with JWA, and negative correlation with Bcl-2. The expression of HSP27 was not detected. Contrast to the cells from APL patient, the expression of JWA need not be activated by ATRA in advance. In this study, we also exposed HL-60 cells in higher dose of Ara-C (20 ng/ml), and JWA expression underwent opposite trend comparing with in lower dose of Ara-C (10 ng/ml). It is concluded that JWA may play double important roles in regulating ATRA and TPA-induced differentiation and apoptosis in leukemic cells. The JWA expression had a negative correlation between induction and cytotoxic response. The difference of JWA expressions between HL-60 cell and ANLL patient cells would be involved in different leukemia pathogenesis.
Antineoplastic Agents ; pharmacology ; Blotting, Western ; Cell Differentiation ; drug effects ; Cytarabine ; pharmacology ; HL-60 Cells ; HSP27 Heat-Shock Proteins ; HSP70 Heat-Shock Proteins ; biosynthesis ; genetics ; Heat-Shock Proteins ; biosynthesis ; genetics ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Neoplasm Proteins ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tetradecanoylphorbol Acetate ; pharmacology ; Time Factors ; Tretinoin ; pharmacology

Objective To determine factors affecting postoperative survival of elderly patients with hepatocellular carcinoma (HCC). Methods A retrospective analysis of consecutive 54 elderly patients undergoing hepatectomy for hepatocellular carcinoma from Jan 1995 to Dec 2002 was performed. The data were analyzed by Kaplan-Meier method and Cox regression. Results Univariate analysis and Cox regression showed Child Pugh grading, vessel invasion, satellite nodule formation, Edmondson Steiner grading, intrahepatic recurrence and distant metastasis all related to postoperative overall survival or disease-free survival of the elderly with hepatocellular carcinoma (all P

Objective: To analyze and review the epidemiology , clinical features , treatment and prognosis of brucellosis ,in order to raise awareness of brucellosis in clinic .Methods:Eight clinical cases of brucellosis in Changzheng Hospital from 2010‐2015 were analyzed retrospectively ,and the clinical treatment experiences were concluded .Results:Five patients had history of epidemic exposure ,the epidemic exposure history of 3 cases were unknown .All patients had fever , fatigue and hyperhidrosis ,which 5 cases were accompanied by arthritis ,orchitis ,lymphadenectasis ,or migratory myalgia .The percentages of neutrophils in 8 cases were normal or relatively low ,and the erythrocyte sedimentation rate (ESR) was elevated in all patients ,in addition ,some indexes of liver function were elevated in 3 cases .All patients were cured by anti‐infective therapy , one patient who re‐contacted with goat was recrudescence .Conclusions: Brucellosis shows various clinical manifestations .Patients in non‐epidemic areas who have long‐term fever ,fatigue ,sweating or joint and muscle pain should be guarded against brucellosis ,and provide patients with early diagnosis and anti‐infective therapy .

OBJECTIVETo construct a rheumatoid arthritis-specific full-length fully human mammalian display antibody libraries.

METHODSPeripheral blood lymphocytes were isolated from patients with rheumatoid arthritis. The repertoires of kappa light chain (LCκ) and heavy chain variable region (VH) of the antibodies were amplified by RT-PCR. The amplified LCκ and VH genes were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO-10 strain to construct the rheumatoid arthritis-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length fully human antibody expressed on the surface of 293T cells were analyzed by flow cytometry.

RESULTSThe libraries of rheumatoid arthritis-specific antibody LCκ and heavy chain (IgG1) were constructed. The expression of full-length fully human antibody on the surface of 293T cells was confirmed by flow cytometry. With the rates of correct LCκ and heavy chain sequence insertion reaching 80% and 60%, respectively, as shown by DNA sequence analysis of the randomly selected clones, the libraries showed an expressible combinatory diversity of 6.13×10(10).

CONCLUSIONThe constructed libraries provide a useful platform for screening rheumatoid arthritis-specific antibodies.

Amino Acid Sequence ; Animals ; Antibodies ; genetics ; immunology ; Antibody Specificity ; Arthritis, Rheumatoid ; immunology ; Cell Surface Display Techniques ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; HEK293 Cells ; Humans ; Immunoglobulin G ; biosynthesis ; genetics ; Immunoglobulin Heavy Chains ; biosynthesis ; genetics ; Immunoglobulin kappa-Chains ; biosynthesis ; genetics ; Lymphocytes ; immunology ; metabolism ; Molecular Sequence Data ; Peptide Library ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Transfection

OBJECTIVETo sort CD133(+) subset cells in human gastric cancer (GC) and to identify their tumor initiating cell-like properties.

METHODSThe tissues of GC and normal tissues adjacent to GC were obtained from 50 patients. Samples were stained for CD133 by immunohistochemistry. Likewise, assessments of CD133 were undertaken by Western blot. Flow cytometry was used to determine the proportion of CD133(+) cells in four GC cell lines therein the KATO-III was sorted by magnetic activated cell sorting (MACS) method. The growing characteristics and the tumorigenic ability of CD133(+) cells were evaluated in vitro and in vivo. Meanwhile, the growth of single cells in suspension culture was observed and expression of stem cell-specific marker were determined using reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe expression of CD133 was demonstrated on the cell membranes in the mucosa and submucosa of primary GC, which were higher than those in the normal gastric tissues adjacent to cancer (P<0.05). Four GC cell lines including KATO-III, SGC-7901, AGS and MKN-45 were found to contain (28 ± 2)%, (17 ± 2)%, (6 ± 2)%, and (4 ± 2)% of CD133(+) cells respectively. In addition, the purity of CD133(+) cells isolated from KATO-III by MACS was (91 ± 3)% and up to(95 ± 2)% after 1-week culture. CCK-8 detection showed that population doubling time of the CD133(+) cells was (21 ± 3)h, significantly shorter than that of the CD133(-) cells[(40 ± 8)h, P<0.05]. Notably, there was a remarkable difference of tumor formation rate between CD133(+) cells (100%), non-sorted cells (80%), and CD133(-) cells(0). The average mass and volume of tumor in group of CD133(+) cells was larger and heavier than those in non-sorted cells (P<0.05, P<0.05). Furthermore, the single cell proliferated well, formed the big sphere and semi-quantitative RT-PCR showed expression of stem cell markers such as Oct-4, Nanog, Sox-2, Musashi-1 and EGFR.

CONCLUSIONSCD133 protein expression in primary lesions is higher than those in the normal gastric tissues. CD133(+) subset cells can be isolated, purified, and amplified in human GC, and possess some properties including the ability of self-renewal, proliferation, and higher tumorigenic ability in vivo and can express some stem cell markers.

AC133 Antigen ; Adult ; Aged ; Aged, 80 and over ; Animals ; Antigens, CD ; metabolism ; Cell Line, Tumor ; Female ; Glycoproteins ; metabolism ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Middle Aged ; Neoplastic Stem Cells ; metabolism ; Peptides ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; Xenograft Model Antitumor Assays

Objective To understand the risk situation of imported schistosomiasis and its main influencing factors in Shang-hai City from 2005 to 2015. Methods A retrospective survey was performed,and the database was established based on the da-ta collection of imported schistosomiasis,Oncomelania hupensis snail situation and mobile population in Shanghai City from 2005 to 2015. Results From 2005 to 2015,the mobile population in Shanghai City increased by 123.92％,which reached about 9816500,and 54.70％of them were located in the inner suburban districts. The accumulated areas with snails of 7.13 hm2 were found in 16 towns of 4 outer suburbs(excluding Chongming District). A total of 23 cases of imported schistosomiasis were found in Shanghai City,and the number of the imported schistosomiasis cases was on the downward trend(rs=-0.782,P=0.004). From 2005 to 2009,11 out of 13 imported schistosomiasis cases(84.62％)were distributed in the inner suburban dis-tricts where no O. hupensis snails were found at the same time. From 2010 to 2015,9 out of 10 imported schistosomiasis cases (90％)were distributed in the outer suburban districts where O. hupensis snails were found at the same time. Conclusion The number of imported schistosomiasis cases in Shanghai is on the downward trend. However,the threat of imported schistosomiasis to the prevention and control of schistosomiasis in Shanghai City should not be ignored.

Objective To investigate the iodine nutrition status of the target population living in the areas with low coverage rate of iodized salt and to provide a basis to prevent and control iodine deficiency. Methods The investigation was carried out in the areas with coverage rate of iodized salt lower than 80%, including 7 counties (city, district). Three townships(sub-district office) were sampled in each county and two elementary schools in each township(sub-district office). Urinary iodine level was measured for 40 children aged from 8 to 10 years old in each elementary school. Drinking water iodine was collected and determined in their living villages. Twenty salt samples were tested for iodine in salt from 20 house which had fertile women, and urine iodine of 10 fertile women were tested in each village. Results Sixty-two water samples were determined and the water iodine was ranged from 5.8 to 272.7 μg/L, of which 3 water samples were equal and more than 150 μg/L. Eight hundred and seventy-two salt samples were collected. The coverage rate of iodized salt was 70.74%(585/827) and the coverage rates were below than 80% in 5 counties (city, district). A total of 1660 children' urine samples were collected, the content of urine iodine ranged from 10.0 to 1088.0 μg/L and the urine iodine median was 173.7 μg/L. Four hundred and thirty-seven urine samples were collected from the fertile women and the urine iodine median was 179.1 μg/L. The iodine level of children and women was the highest in Dongguang County(251.8,273.8 μg/L) while that of Hejian County (130.8,118.7 μg/L) was the lowest. Conclusions Although the iodine nutrition of children and fertile women is appropriate in areas with low coverage rate of iodized salt, we presume from the results that the possibility of iodine deficiency in pregnant and lactating women exists in Hejian and Anping.

OBJECTIVETo study the feasibility and validity of multi-modal serial therapy for primary liver cancer in senile patients.

METHODS153 senile primary liver cancer patients (>or= 60 years) were given multi-modal serial therapy from June 1993 to December 2000. Hepatectomy was performed in 37, deep cryosurgery in 32 and non-operative therapy in 84 (intervention as chief therapy in 81, combined local and intervention therapy in 3). The multi-course intervention therapy was given postoperatively in hepatectomy and cryosurgery groups, while bioimmunotherapy and traditional Chinese medicine were used in all groups.

RESULTSThe 1-, 3- and 5-year survival rates in the hepatectomy group were 78.4%, 46.4% and 35.7%, without operative mortality. The 1- and 3- and 5-year year survival rates in the cryosurgery group were 64.5%, 40.9% and 25.0% with mortality of 3.1%. Among patients with non-operative therapy, the 1- and 3- and 5-year year survival rates in intervention group were 47.5%, 23.5% and 4.3%. The operative mortality was 1.2%. The 3 patients who received combined local and intervention therapy have survived for 2.5, 3.8 and 7.1 years.

CONCLUSIONMulti-modal serial therapy with surgical treatment as the chief means, being precise in the effect and good in safety, is feasible and valid for primary liver cancer in senile patients.

Aged ; Aged, 80 and over ; Antineoplastic Agents ; therapeutic use ; Carcinoma, Hepatocellular ; pathology ; therapy ; Chemoembolization, Therapeutic ; Combined Modality Therapy ; Cryosurgery ; Feasibility Studies ; Female ; Hepatectomy ; Humans ; Infusions, Intra-Arterial ; Liver Neoplasms ; pathology ; therapy ; Male ; Middle Aged ; Neoplasm Staging ; Survival Rate