OBJECTIVETo clarify the gene type of Orientia tsutsugamushi (Ot) from Shandong province.

METHODSNested-polymerase chain reaction (nPCR) was used to identify the gene type of 23 isolated Ot strains, 2 pools of homogenized leptotrombidium (L.) scutellare, 10 blood specimens of scrub typhus patients, and at the same time to compare with the international reference strains Gilliam, Karp, Kato. Sequencing analysis of the Sta56 gene was also used to further identify the precise gene types.

RESULTSOf the 35 samples, 33 had the same products in the amplification of template Ot-DNA. They all belonged to Kawasaki strains endemic in Japan while 2 (FXS4 and LHGM2 strain) belonged to Karp strains. The Sta56 gene sequence homologies to Japan Kawasaki strain of the 2 representative strains (B-16 and FXS2 strain) of the 33 samples were 94.22%, 95.21% respectively, but they were less than 75.87% to other prototype strains; The homologies to Karp strain of FXS4 and LHGM2 strain were 83.03%, 96.45% respectively. B-16 and FXS2 strain were designated as of types strain Japan Kawasaki, FXS4 and LHGM2 as Karp strain.

CONCLUSIONThe results indicated that the dominant Ot strains in Shandong Province were similar to Kawasaki strains, but Karp strains also existed.

Animals ; DNA, Bacterial ; genetics ; Genotype ; Humans ; Mice ; Orientia tsutsugamushi ; genetics ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Scrub Typhus ; epidemiology ; microbiology ; Sequence Homology ; Serotyping

OBJECTIVETo study the pattern of lymph node spread in papillary thyroid carcinoma (PTC) with clinically negative node (cN0).

METHODSA total of 106 patients with cN0 PTC who underwent total or subtotal thyroidectomy plus unilateral or bilateral lateral neck dissection (LND, level II-V or level I-V) at West China Hospital of Sichuan University between April 2004 and August 2010 were analyzed retrospectively.

RESULTSThe lateral neck lymph node metastasis in cN0 PTC was significantly associated with sex (male, P = 0.007), tumor stage (T3/T4, P = 0.006), tumor size (> 1 cm, P = 0.014) and the number of positive central lymph nodes (≥ 2, P < 0.001), but not with age and multifocal tumor. Level III (47/116, 40.5%) was the most prevalent metastatic site, followed by level IV (41/116, 35.3%), level II (18/116, 15.5%) and level V (2/29, 6.9%). Of the cases with lymph node metastases in level III and IV, 89.8% (79/88) of primary thyroid tumors existed in the lower and middle sites of the thyroid lobes, while in the cases with lymph node metastases in level II, 77.8% (14/18) of primary thyroid tumors in the upper sites of the thyroid lobes, and 83.3% of cases with level II metastases were accompanied with level III metastases. Two cases with level V metastases were accompanied with metastases in levels II, III and IV.

CONCLUSIONSLND should be considered for cN0 PTC in male, with T3/T4 lesions and positive central lymph nodes ≥ 2, and the range of dissection should include level III and IV. Dissection of level II should be considered in cN0 PTC with primary tumor localized in the upper site of the thyroid lobe or with level III metastasis. Dissection of level V should be considered at present of metastases in level II, III, and IV. For cN0 PTC with tumor size < 1 cm, confined to the thyroid and without lymph node metastasis in the central compartment, LND is not recommended.

Adolescent ; Adult ; Aged ; Carcinoma ; pathology ; Carcinoma, Papillary ; Child ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Neck Dissection ; Neoplasm Staging ; Retrospective Studies ; Thyroid Neoplasms ; pathology ; Young Adult

Different cytokines are needed in the course of culturing cells to do adoptive immunotherapy. This study was aimed to investigate the differentiation directions of lymphocytes and related gene expression characteristics after combined stimulation of lymphocytes by different cytokines or EBV antigen peptide combined with cytokines. The experiment was divided into 4 groups. The levels of total T lymphocytes (CD3(+)), T helper lymphocytes (CD3(+)CD4(+)), cytotoxic T-lymphocyte (CD3(+)CD8(+)), memory T cells (CD3(+)CD8(+)CD45RO(+)), naive T cells (CD3(+)CD8(+)CD45RA(+)), Th2 cells (CD3(+)CD30(+)), B cells (CD19(+)), NK cells (CD56(+)), naive T regulatory cells (CD4(+)CD25(+)), precise T regulatory cells (CD4(+)CD25(+)FOXP3(+)) were detected by flow cytometry. The expression levels of house-keeping gene (mad1, pten), T helper cells transcriptional regulatory gene t-bet (Th1), gata3 (Th2), cytokine IFN-γ(Th1), IL-4(Th2) were detected by using RT-PCR. The results showed that CTL in EBV polypeptide group were dominant cells with certain clinical effects. Comparison of result of EBV polypeptide group with other 3 different cytokine stimulating groups demonstrated that EBV antigen peptide had much more effects on stimulating CTL generation. The expression of IFN-γ gene was significantly increased; the T helper differentiation-related gene t-bet, gata3 also increased evidently, while expression change of house-keeping gene mad1 and pten were not evident. Addition of different cytokines and antigen peptides in culture may be much more effective on stimulating CTL generation. It is concluded that specific CTL can be obtained by using the lymphocytes co-cultured with EBV and cytokines, and the different cytokines play different roles in cell differentiation.
Cells, Cultured ; Cytokines ; immunology ; metabolism ; Epstein-Barr Virus Nuclear Antigens ; genetics ; Flow Cytometry ; Humans ; Immunotherapy, Adoptive ; Lymphocyte Count ; Lymphoma, Extranodal NK-T-Cell ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; immunology

Aliquots of venous blood from healthy donor were collected in plastic blood storage bags with ACD, GMA or antioxidant solution (superoxide dismutase, SOD), respectively, and stored at 4 degrees C. After storage for varying periods, the parameters of the blood were detected in the blood samples. Results showed that the parameters of the blood stored at 4 degrees C for 75 days in SOD group were following: the recovery of RBC-Hb was 87.2%, plasma-Hb (mg/L) was 193.2, P50 (mmHg) was 34.0 (normal value was 33.1); deformability (DImax) was 0.2413 (74.3% of normal value). There was no evident hemolysis, color change, air bubble and clots. It was concluded that human RBC stored at 4 degrees C for 75 days with SOD solution, recovery of levels of RBC-Hb and plasma-Hb were accorded with the requirements of "Basic Demands of Blood Station" in China.
Antioxidants ; pharmacology ; Blood Preservation ; methods ; Cold Temperature ; Erythrocyte Deformability ; drug effects ; Erythrocytes ; drug effects ; metabolism ; Free Radical Scavengers ; pharmacology ; Hemoglobins ; drug effects ; metabolism ; Humans ; Superoxide Dismutase ; pharmacology ; Time Factors