Objective To evaluate the performance of Mindray TSH chemiluminescence immunoassay ,including imprecision , limit of detection ,functional sensitivity ,interference ,cross-reaction ,and calibration consistency .The purpose was to confirm Mind-ray TSH assay meets the criteria of IFCC standardization and harmonization of thyroid functional tests .Methods The CLSI guide-lines defined in EP documents have been followed to measure the limit of detection ,functional sensitivity ,specificity ,imprecision and calibration consistency of Mindray TSH immunoassay on CL2000i system .Results Limit of detection was 0 .001 5 μIU/mL ,func-tional sensitivity was 0 .013 μIU/mL ,the correlation coefficient of linearity was 0 .999 in the range of 0-98 .95 μIU/mL .Mindray TSH calibrator C0 spiked with luteinizing hormone (LH) up to 500 mIU/mL ,follicle stimulating hormone (FSH) up to 500 mIU/mL ,or human chorionic gonadotropin (HCG) up to 200 000 mIU/mL detected no cross reactivity (detected TSH was less than 0 .2μIU/mL) .Hemoglobin up to 500 mg/dL ,bilirubin up to 10 mg/dL ,triglycerides up to 1 800 mg/dL ,and protein up to 10 g/dL showed -6 .91% ~8 .60% bias in TSH measurement .The total imprecision of two controls (high and low levels) and three serum specimens were in the range of 3 .24% and 5 .34% .Calibration consistency which had been demonstrated with high and low controls were measured between -6 .58% and 5 .26% .Conclusion The performance of Mindray thyroid-stimulating hormone assay meets the criteria for IFCC standardization and harmonization of thyroid function tests .

To enhance the immunogenicity of DNA and adenoviral vector vaccines expressing HIV-1 subtype B gp160, human interleukin 15 (hIL15) DNA adjuvant (pVR-hIL15) was constructed. BALB/c mice received DNA prime/protein boost immunization with pVR-HIVgp160/Ad5-HIVgp160 alone or combined with pVR-hIL15. Cellular and humoral immune responses were evaluated by IFN-gamma enzyme-linked immunosorbent spot assay and enzyme-linked immunosorbent assay, respectively. Compared with those immunized with vaccines alone, the mice immunized with vaccines combined with pVR-hIL15 had significantly increased specific cellular response and antibody titer (P < 0.05). It suggests that the IL15 DNA adjuvant can enhance the immune responses induced by prime-boost regimen using DNA and adenoviral vector encoding HIV-1 subtype B gp160.
Adenoviridae ; genetics ; Adjuvants, Immunologic ; Animals ; Antibodies, Viral ; immunology ; Antibody Specificity ; Female ; Genetic Vectors ; genetics ; HIV Envelope Protein gp120 ; immunology ; HIV Envelope Protein gp160 ; genetics ; immunology ; HIV Envelope Protein gp41 ; immunology ; Humans ; Immunity, Cellular ; Immunity, Humoral ; Interleukin-15 ; genetics ; Mice ; Mice, Inbred BALB C ; Vaccines, DNA ; genetics ; immunology

Objective To explore the mechanism of HPV infection in carcinogenesis and progression of colon cancer. Methods Human colon cancer cells, HCT116 (with wild-type p53) and SW480 (with mutant-type p53), were transfected by HPV16 E6E7 oncogenes using a recombinant adeno-associated virus vector system. The transfection efficiency was determined by flow cytometry. The expression of HPV16 E6 genes was determined by Western blot. The cell proliferation and cell cycle was studied by MTT method and flow cytometry. Results Western blot confirmed the expression of E6 gene in colon cells that were infected by rAAV-E6E7. The population doubling time of HCT116 cell, which was more than 48 hours at control group, decreased to 33 hours. HPV16 E6E7 increased cell percentage of S phase and decreased cell percentage of G0/G1. The population doubling time of SW480 cell was 77.06% decreased and the OD540 was 47.18% increased with interference of HPV16 E6E7 gene. Conclusion HPV16 E6E7 oncogene precipitates the proliferation and positively controls cell cycle of HCT116 and SW480 human colon cancer cells. HPV infection may closely relate to the carcinogenesis and progression of colon cancer.

Objective To investigate the relationship between HPV infection and human colorectal carcinogenesis. Methods Colorectal carcinoma specimens from 72 Chinese patients were studied. DNA extracted from colorectal tissue was screened for HPV L1 by polymerase chain reaction (PCR), HPV subtype 6,11, 16, and 18 were detected by PCR using specific primers and in situ hybridization using specific probe. Results Twenty-four specimens out of 72 (33%) colorectal cancer were HPV L1 positive. The normal colorectal mucosa was HPV L1 negative. The location, infiltration and metastasis of colorectal carcinoma were all significantly related with HPV infection. The predominant HPV subtype was HPV 16,which was found in 58% of all HPV-positive colorectal carcinoma. Conclusion The presence of HPV DNA suggests that HPV may be involved in the carcinogenesis of colorectal carcinoma. HPV infection is closely related with the malignant potential of colorectal cancer.

Objective To evaluate the effects of rigorous surveillance and retroperitoneal lymph node dissection (RPLND) in the treatment of low-risk patients with clinical stage Ⅰ nonseminomatous germ cell testicular tumors (NSGCT) after radical orchiectomy.Methods The data of 71 patients with clinical stage Ⅰ NSGCT were analyzed retrospectively in Hunan Provincial Tumor Hospital,Xiangya Third Hospital of Central South University and Hunan Provincial People's Hospital between Feb,2001 and Apr,2012.Excluding lymphatic and vascular invasion,percentage of embryonal carcinoma＞50％ and increasing tumour markers (AFP/β-HCG) following orchiectomy,46 low-risk patients out of 71 patients with clinical stage Ⅰ NSGCT were selected and divided into rigorous surveillance group (30 cases) and RPLND group (16 cases) according to different therapeutic methods after radical orchiectomy.Univariate analysis was used to confirm variables associated with disease progression,and the disease free survival rates (DFSR) were compared by using Kaplan-Meier analysis.Results Five cases were lost,and 41 cases were followed up for an average of 61 months (range,15-147 months),with 58 months in rigorous surveillance group (range,19-147months) and 65 months in RPLND group (range,15-144 months).The survival rate was 100％ in 2 groups.The DFSR was 89％ (24/27) and 86％ (12/14),respectively,and there was no significant difference between the 2 groups (x2 =0.08,P=0.78).The DFSR was 83％ in patients with small amout of embryonal (percentage of embryonal carcinoma ＜ 50％),and 92％ in patients without embryonal carcinoma,and there was no significant difference between the 2 groups (x2=1.07,P=0.30).Also there was no significant difference between the patients less than 15 years and patients more than 15 years (x2=1.59,P =0.21).Conclusions There is no significant difference in recurrence rate and DFSR between rigorous surveillance group and RPLND group.Low-risk patients with clinical stage Ⅰ NSGCT may achieve satisfactory prognosis with surveillance after radical orchiectomy.

Boronic Acids ; therapeutic use ; Bortezomib ; Humans ; Male ; Middle Aged ; POEMS Syndrome ; therapy ; Peripheral Blood Stem Cell Transplantation ; Pyrazines ; therapeutic use ; Transplantation, Autologous

Humans ; Occupational Exposure ; Occupational Health ; Risk Assessment ; Workplace

OBJECTIVETo investigate the dynamic changes of neurofilament contents in rat's spinal cord induced by 2, 5-hexanedione (2, 5-HD), and explore the molecular mechanism of n-hexane neuropathy.

METHODSMale Wistar rats were administered at a dosage of 400 mg/kg/day 2, 5-HD for 2, 4 and 8 weeks respectively. HD-induced neurological defects were detected and quantified using gait score, and the relative lev-els of NF-H, NF-M, and NF-L in spinal cords of rats were determined by Western Blotting.

RESULTSExposure to 2, 5-HD produced progressive gait abnormalities, which suggested that the rat model of 2, 5-HD-induced neurotoxicity was established successfully. Western-Blotting results showed that NFs content in spinal cord demonstrated a progressive decline as the intoxication continued. In the supernatant fraction, compared to the controls, NF-H con-tent decreased by 15.7%, 57.0%, and 58.0% respectively after 2, 4, and 8-week treatment with 2, 5-HD (P < 0.01); accordingly, NF-M decreased by 36.0%, 61.3%, and 65.2% respectively (P < 0.01); NF-L decreased by 20.8%, 43.9%, and 44.3% respectively (P < 0.01). In the pellet fraction, the contents of NF-H in groups of 4 and 8 weeks' exposure to HD decreased by 35.6% and 43.2%, respectively (P < 0.01), and those of NF-L decreased by 26.4% and 42.1%, respectively (P < 0.01) when compared to the control. Further-more, NF-M contents in groups of 2, 4 and 8 weeks' exposure decreased by 23.3%, 33.9%, and 63.7% respectively (P < 0.01). The NFs level in spinal cords was highly correlated with gait abnormality of treated rats as the intoxication went on. Multiple correlation coefficients of NF-H, NF-M, and NF-L content with gait score of HD-treated rat were 0.8912, 0.9282 and 0.8981 (P < 0.01) respectively.

CONCLUSIONThe declines of NFs are high-ly related to neurobehavioral abnormality of 2, 5-HD-treated animals, and involved in the development of n-hexane neuropathy.

Animals ; Disease Models, Animal ; Gait ; drug effects ; Hexanones ; toxicity ; Male ; Neurofilament Proteins ; metabolism ; Rats ; Rats, Wistar ; Spinal Cord ; drug effects ; metabolism

OBJECTIVETo investigate the effect and explore the mechanism of Bushen Antai Recipe (BAR) on pregnancy rate and number of implantation site in blastocyst implantation dysfunction (BID) mice induced by indomethacin.

METHODSPregnant mice were divided into 3 groups randomly: the normal group, the model group and the BAR group. Tap water was given orally to the rats in the normal and model groups, and BAR to the rats in the BAR group from the first day of pregnancy for 5 or 8 days; on the 3rd and 4th day dissolvent was injected subcutaneously twice per day in the normal group, while indomethacin (4.33 mg/kg) was injected subcutaneously twice per day in the other two groups to establish implantation dysfunction model; serum estrogen (E) and progesterone (P4) levels were detected on the 5th and 8th day; the pregnancy rate and number of implanted site was observed and the receptors of E and P4 in endometrium of uterus were examined by immunohistochemistry on the 8th day.

RESULTSThe pregnancy rate and number of implanted site was 27.3% and 5.3 +/- 0.7 respectively in the model group, significantly lower than those in the normal group (90.9%, 13.3 +/- 2.8), and the BAR group (72.7%, 10.7 +/- 2.2, P < 0.05). Serum E level was higher in the BAR group than that in the model group on the 5th and 8th day, and even higher than that in the normal group on the 8th day; serum P4 level was lower in the model and BAR groups than that in the normal group on the 5th day (P < 0.01), but higher in the BAR group than that in the model group on the 8th day. Immunohistochemical observation showed that expressions of E and P4 receptor increased remarkably in the BAR group than those in the model group.

CONCLUSIONBAR increases the pregnancy rate and number of implanted site of indomethacrne induced BID mice through regulating E and P4 levels and enhancing the expressions of their receptors in the endometrium.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Embryo Implantation ; drug effects ; Embryo Implantation, Delayed ; drug effects ; Estrogens ; blood ; Female ; Indomethacin ; Mice ; Pregnancy ; Progesterone ; blood ; Random Allocation

OBJECTIVETo compare the effects of carvedilol, metoprolol and propranolol on myocardial gap junction (GJ) structure in rat with myocardial ischemia and reperfusion injury.

METHODSRats were divided randomly into five groups: sham operation group (SO), myocardial ischemia and reperfusion group (IR), IR + carvedilol group (CV), IR + metoprolol group (MT), and IR + propranolol group (PP). The left anterior descending branch was ligated for 30 minutes and reperfused for 4 hours (IR). After 4 h reperfusion, the distribution and composition of gap junctional connexin 43 (CX43) were observed by immunofluorescence and laser scanning confocal microscopy (LSCM), and the quantification of CX43 was measured by LSCM.

RESULTCompared with SO group, IR resulted in abnormal distribution and composition of CX43-GJ and the impairment of CX43-GJ was significantly attenuated by CV, MT and PP treatments with the best effect observed in CV group (P<0.05 vs. MT and PP).

CONCLUSIONThese results suggest that beta-blockers, especially, carvedilol, could significantly attenuate IR induced CX43-GJ impairment.

Adrenergic beta-Antagonists ; pharmacology ; Animals ; Connexin 43 ; metabolism ; Disease Models, Animal ; Gap Junctions ; drug effects ; Male ; Myocardial Reperfusion Injury ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley