Objective:To express fusion protein of mouse thymic stromal lymphopoietin (TSLP) and HIV-1 gp120BAL V1/V2 subdomain in 293F cell.Methods:Full length of the V1V2 sequence from BAL isolate was fused with the C-terminus of mouse thymic stromal lymphopoietin (TSLP) and sub-cloned into pCEP-Pu vector to construct the eukaryotic expression plasmid-pCTV1V2BAL.The recombinant plasmid was confirmed by enzyme digestion and sequencing , then transfected into 293 F cells using PEI as a transfection reagent .The fusion protein was purified by metal chelate affinity chromatography and characterized by SDS -PAGE and Western blot . The epitopes of V1/V2 in fusion protein were identified by ELISA .Results:The SDS-PAGE and Western blot results showed that there were highly heterogeneous glycoprotein bands at the site between 35 kD and 55 kD, which reacted with anti-mTSLP rabbit polyclonal antibody and anti-His tag mouse monoclonal antibody .The ELISA analysis showed that antibodies to V 1/V2BAL existed in the sera of HIV-1 positive patients.Conclusion:The mTSLP-V1/V2 fusion protein was successfully expressed in 293F cells, which may be useful for HIV-1 vaccine research .

Adolescent ; Adolescent Development ; Child ; Child Development ; China ; epidemiology ; Female ; Humans ; Male ; Obesity ; epidemiology ; Prevalence

BACKGROUNDNominal atrioventricular (AV) interval in dual chamber pacemaker (DDD) is not the best AV delay in the majority of patients with atrioventricular block. To find a simple method for optimizing AV delay adjustment, we assessed surface electrocardiography (ECG) for optimizing AV delay during dual chamber pacing.

METHODSDDD pacemakers were implanted in 46 patients with complete, or almost complete, AV block. Optimal AV delay was achieved by programming an additional delay of 100 ms, to the width of intrinsic P wave or to the interval between pacing spike to the end of P wave on surface ECG. Left ventricular (LV) end diastolic and end systolic volumes, ejection fraction and diastolic parameters were measured by Doppler echocardiography during both nominal and optimal AV delay pacing.

RESULTSCompared to nominal AV delay setting, LV end diastolic volume increased [to (53.2 +/- 11.3) ml from (50.2 +/- 10.2) ml, P < 0.05], end systolic volume decreased [to (26.1 +/- 9.0) ml from (27.9 +/- 8.2) ml, P < 0.05] during adjusted AV delay pacing, resulting in an increase in LV ejection fraction [to (68.2 +/- 5.3)% from (64.5 +/- 4.3)%, P < 0.05]. LV diastolic filling and isovolumic relaxation time were not significantly changed.

CONCLUSIONOptimization of AV delay by surface ECG is a simple method to improve LV systolic function during dual chamber pacing.

Adult ; Aged ; Aged, 80 and over ; Atrioventricular Node ; physiopathology ; Cardiac Pacing, Artificial ; methods ; Electrocardiography ; methods ; Female ; Heart Block ; physiopathology ; therapy ; Humans ; Male ; Middle Aged ; Time Factors

OBJECTIVETo investigate the clinicopathological characteristics, differential diagnosis and prognosis of testicular cellular fibroma.

METHODSWe comprehensively analyzed the clinical presentation, histomorphology and immunohistochemistry of a case of testicular cellular fibroma, reviewed the relevant literature, and discussed its pathological features and differential diagnosis.

RESULTSA 30-year-old man presented with complaint of discomfort and painless enlargement in the right testis. The tumor was found to be a testicular fibroma characterized by a solid, thickly or thinly encapsulated, circumscribed and gray-white mass. Microscopically, fusiform cells were arranged into a storiform and herringbone pattern or fascicles. The tumor exhibited a great deal of cellularity and no nuclear polymorphisms, with a mitotic rate of 0-1/10 HP. Immunohistochemistry showed that the tumor cells were positive for Vimentin, patchily positive for S-100 and SMA, but negative for Desmin, alpha-inhibin, CD34 and CD99. The positive rate of Ki-67 was less than 1%.

CONCLUSIONTesticular cellular fibroma is a rare testicular sex cord stromal tumor, pathologically resembling its ovarian counterpart. It can be distinguished from other testicular spindle cell tumors by morphology and immunohistochemical staining. For the treatment of testicular cellular fibroma, surgical resection often has a good prognosis.

Adult ; Fibroma ; pathology ; Humans ; Immunohistochemistry ; Male ; Testicular Neoplasms ; pathology

Objectives: To assess the relationship of variation of blood pressure and neurological deterioration (ND) in ischemic stroke patients. Methods: We recruited patients with the fi rst-ever ischemic stroke at a teaching hospital. The National Institutes of Health Stoke Score (NIHSS) of each patient was monitored for 2 months. ND was defi ned as an increase of ≥ 2 points in NIHSS during the fi rst 7 days after stroke. Blood pressure was measured every 6 hours for fi rst 7 days. We analyzed blood pressure data in the fi rst 36 hours to study the relationship between variation of blood pressure and ND. Successive variation of systolic (svSBP) and diastolic (svDBP) blood pressure was calculated as svSBP= |SBPn+1 – SBPn | and svDBP= |DBPn+1 – DBPn | respectively. The largest svSBP in the fi rst 36 hours of hospitalization or before ND was defi ned as maximum variation of systolic blood pressure (maxvSBP). Then, the mean variation of systolic (mvSBP) and diastolic (mvDBP) blood pressure was calculated as mvSBP= svSBP/N and mvDBP= svDBP/N respectively. Results: A total of 121 patients were included in this study, and 38 of them had ND. The mvSBP was higher in the ND Group (17.9±8.4 mmHg vs. 13.7±4.4 mmHg, p=0.006) but the difference in mvDBP did not reach statistical signifi cance (9.8±3.5mmHg vs. 8.6±3.0 mmHg p=0.06). The ND Group had a larger maxvSBP (35.2±17.2 vs. 27.6±11.6 mmHg, p =0.01), which was more frequently over 30mmHg than that in the stable group (P=0.02). Conclusions: A large svSBP is associated with an increased risk for ND. The study highlights the importance of close monitoring of blood pressure in ischemic stroke patients.

OBJECTIVESTo investigate the association between genetic polymorphisms ofX-ray repair crosscomplementing group 1 (XRCC1) codons 194, 280, and 399 and cervical neoplasm susceptibility.

METHODSA community-based nested case-control study was conducted. The study population consisted of women living in Chiayi City, located in southwestern Taiwan, who had received pap smear screening between October, 1999, and December, 2000 (n=32,466). The potential cases were women having lesions greater than cervical intraepithelium neoplasm II (C1N2) reconfirmed by cervical biopsy. The potential controls (case: control=1∶2) were age matched (±2 yrs) and residency matched women who had had normal pap smears. In total, 100 cases (39 C1N2, 12 C1N3, 46 carcinoma in situ (CIS), and 3 invasive cancer) and 196 controls had the information on both questionnaire and data ofXRCC1 polymorphisms.

RESULTSThe frequency ofArg/Arg, Arg/Gln, andGln/Gln in codon 399 among cases and controls was 54% (54/100), 38% (38/100), and 8% (8/100) and 58% (114/196), 37% (73/196), and 5% (9/196), respectively, which were not significantly different. No associations were also observed betweenXRCC1 codon 194 and 280 genotypes and cervical neoplasm. While dichotomized by age (<40 vs. ≥40 yrs), smoking status (active and passive smokers vs. non-smokers), and disease status (C1N2 and C1N3 vs. CIS and invasive cancer), the results remained insignificant.

CONCLUSIONSThe present findings suggest thatXRRC1 codon 194, 280 and 399 genotypes may not influence cervical neoplasm risk in the Taiwanese population.

This study was aimed to evaluate the in vivo antitumor effect of genetically modified myeloma cell vaccine on human myeloma xenografts implanted into nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice. Human immune system was established in NOD/SCID mice by intraperitoneal injection of human peripheral blood lymphocytes (PBLs). After being inoculated subcutaneously with irradiated myeloma cell line sko-007, adenovirally transferred with GFP or p53, granulocyte-macrophage colony-stimulating factor (GM-CSF) and B7-1 genes, huPBL-NOD/SCID mice were challenged by subcutaneous injection of non-transferred sko-007 cells. The results indicated that Ad-p53/GM-CSF/B7-1-infected sko-007 cell vaccination significantly reduced local tumor growth compared with controls. Histopathological and immunohistochemical analysis showed that tumor tissues increasingly displayed diffuse necrosis, mainly caused by apoptosis, accompanied with significant fibroplasias and blood vessel hyperplasia, and human T cells infiltrated into the tumor tissues. It is concluded that transgenic p53, GM-CSF and B7-1 expression produces an immune response against myeloma cells and may be of therapeutic value for multiple myeloma in human being.
Adenoviridae ; genetics ; Animals ; B7-1 Antigen ; genetics ; immunology ; Cancer Vaccines ; immunology ; Genes, p53 ; immunology ; Genetic Vectors ; Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; immunology ; Immunotherapy ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Multiple Myeloma ; immunology ; Neoplasm Transplantation

INTRODUCTIONThere is no single method of measuring insulin resistance that is both accurate and can be easily performed by general researchers. We validate the accuracy of oral glucose insulin sensitivity (OGIS) in the Chinese by comparing the OGIS120 and OGIS180, homeostasis model assessment of insulin resistance (HOMA-IR), and quantitative insulin sensitivity check index (OUICKI) with steady-state plasma glucose (SSPG) in different glucose tolerance subjects.

MATERIALS AND METHODSWe enrolled 515 subjects, aged between 20 and 75 years old, during routine health evaluations. All subjects were divided into normal, obese, impaired fasting glucose (IFG), impaired glucose tolerance (IGT) and type 2 diabetes (T2D) groups. Participants had a 3-hour oral glucose tolerance test (OGTT) and SSPG with an insulin suppression test. The relationships between SSPG and OGIS120, OGIS180, HOMA-IR, and QUICKI were evaluated.

RESULTSThe normal group had the highest OGIS120, OGIS180 and lowest SSPG as compared with the other 4 groups. OGIS180, HOMA-IR and QUICKI in all 5 groups were significantly related to SSPG (r = 0.397-0.621, all P <0.05). OGIS120 in all 5 groups was not significantly related to SSPG (r = 0.003-0.226). Additionally, the r value of OGIS180 against SSPG was not higher than the other 2 insulin sensitivity surrogates from OGTT.

CONCLUSIONSAlthough OGIS180 was more accurate in estimating insulin sensitivity than OGIS120 in the Chinese, it was not superior to the traditional surrogates such as HOMA-IR or QUICKI.

Adult ; Aged ; Case-Control Studies ; China ; Female ; Glucose Tolerance Test ; methods ; Humans ; Insulin Resistance ; Male ; Middle Aged ; Prediabetic State ; diagnosis ; Young Adult

[Objective]To investigate the effect of KIF23 gene expression on the proliferation,migration and invasion of human hepatocellular carcinoma HepG2 cells in vitro,and to explore the possible mechanism.[Methods]The KIF23 siRNA was transfected into HepG2 cells by lipofectamine 3000.The expression of KIF23 mRNA and protein in HepG2 cells was de-tected by qRT-PCR and Western blot.The effect of silencing KIF23 on the proliferation of HepG2 cells was studied by CCK-8 assay and plate clone formation assay.The tumor cell abilities of migration and invasion after transfection were measured by scratch assay and Transwell assay.The expression of protein kinase B(PKB/Akt)and phosphorylated Akt(p-Akt)protein in HepG2 cells transfected with KIF23-siRNA2 was detected by Western blot.[Results]KIF23-siRNA could effectively si-lence the expression of KIF23 mRNA and protein in HepG2 cells(P<0.01).The results of CCK-8 assay,plate clone forma-tion assay,scratch assay and Transwell assay demonstrated that the cell proliferation,migration and invasion ability of the KIF23-siRNA2 interference group were significantly inhibited,compared to the negative control group and the blank control group(P<0.05).The expression level of total Akt protein in HepG2 cells was not changed,but the expression level of phos-phorylated Akt protein was down-regulated(P<0.05).[Conclusions]KIF23 may promote the proliferation,migration and in-vasion of human hepatocellular carcinoma cells by activating Akt signal transduction pathway.KIF23 is expected to be a new target for gene therapy of hepatocellular carcinoma.