Objective To evaluate the two methods for quantitative hepatitis B virus (HBV) DNA test. Methods The Hybrid Capture II system from Digene Co. and Real Time PCR fluorimetry quantitative HBV DNA test kit from PIJI Bio Technical Development Company Ltd. were used to detect sera HBV DNA in sera of patients with chronic hepatitis B. Results With the two quantitative methods, the positive rates of HBV DNA were 98 6% and 94.6% respectively. The concordant rate of these two methods was 93.2%. In assessing the sensitivity of the two kits with a serial of 10 fold diluted patients′s sera, PG kit could test 10 -7 of diluted serum, and HC II could test 10 -6 . The test value of HC II was more accurate in quantitation than PG kit especially in higher titer of HBV DNA. Whereas in the case of low titer of HBV DNA, the deviation of test value increased in both two methods. For monitoring the anti virus effect with quantitative HBV DNA in CHB patients, these two methods had same sensitivity, and the test value of HBV DNA had the same trend of change. Conclusion HC II system and PG test kit were sensitive and reliable, and they were worthy of monitoring the anti virus efficacy and of clinical practice.

OBJECTIVETo explore the therapeutic effect of Chinese medicinal massage with nourishing-Shen and activating-blood manipulation in treating women with climacteric syndrome and its influence on endocrinal function.

METHODSSixty patients were assigned to two groups, the 40 patients in the treated group were treated by Chinese medicinal massage for 20 min once every other day; the 20 patients in the control group were treated by hormone replacement therapy with Premarin 0.625 g, once daily by oral taking. The therapeutic efficacy was evaluated after two-month treatment by changes of serum levels of estradiol (E2), follicular stimulating hormone (FSH), luteinizing hormone (LH), and Kupperman index before and after treatment.

RESULTSKupperman index showed that the total symptom score in the treated group was improved from 30.71 +/- 8.43 scores before treatment to 8.21 +/- 5.14 scores after treatment, with a decrement of 22.50 +/- 8.14 scores, which was higher than that in the control group, from 24.32 +/- 5.44 scores to 5.92 +/- 3.58 scores, with a decrement of 18.40 +/- 4.50 scores, the difference between them was statistically significant (t = 2.52, P = 0.014). The serum level of E2 increased in both groups significantly after treatment, from 20.23 +/- 20.78 ng/L to 54.34 +/- 24.26 ng/L in the treated group (t= -2.73, P = 0.006), and from 16.15 +/- 24.40 ng/L to 40.61 +/- 81.54 ng/L in the control group (t = -1.72, P = 0.086), but the difference between groups was statistical insignificant (t= -1.120, P = 0.263). As for levels of FSH and LH, their decrements in the control group (13.16 +/- 11.29 mlU/mL and 10.37 +/- 9.21 mlU/mL) were larger than those in the treated group (4.92 +/- 4.26 mlU/mL and 0.17 +/- 2.42 mlU/mL), respectively (t = - 2.49, P = 0.013; t = - 2.38, P = 0.017).

CONCLUSIONChinese medicinal massage manipulation could improve the Kupperman index of all the 13 symptoms in women with climacteric syndrome, and increase the E2 level in serum.

Adult ; Estradiol ; blood ; Estrogens, Conjugated (USP) ; therapeutic use ; Female ; Follicle Stimulating Hormone ; blood ; Humans ; Luteinizing Hormone ; blood ; Massage ; Menopause ; Middle Aged

OBJECTIVETo explore the underwater decompression schedule for 100 m Trimix conventional diving operations and evaluate its safety through a simulated rabbits Trimix conventional diving.

METHODSAccording to the Haldane theory, the assumed time units, the classification of tissue compartments, the nitrogen super-saturation safety coefficient and the selection of methods used for the calculation of the simulated 100 m Trimix conventional diving schedule were properly selected, and the calculating method for the dive decompression schedule was thus firmly established. In our experiments, five tissue compartments were selected during the calculation of decompression schedule: 5 min, 10 min, 20 min, 40 min and 75 min, and the nitrogen super-saturation safety coefficient was calculated by 1.6. Eight New Zealand rabbits were performed a simulated 100 m Trimix dive program which was established according to the Haldane theory, and eight rabbits for intact group. The tissues wet/dry ratio and ethology were detected and observed before and after the simulated diving to evaluate the safety of decompression schedule.

RESULTSBy using the developed underwater decompression schedule, abnormal ethology changes in rabbits could not be observed after compression and decompression to the surface; and the tissues wet/dry ratio of simulated diving rabbits had no significant changes compared with the intact group (P > 0.05).

CONCLUSIONThe decompression schedule calculated by Haldane theory seemed to be safe and reliable, the diving breathing gas concentration did not cause oxygen toxicity and nitrogen narcosis among the dive rabbits, and dive efficiency was greatly improved by using enriched oxygen gas in UPTD safety range during decompression.

Animal Experimentation ; Animals ; Decompression ; Diving ; Helium ; Nitrogen ; Oxygen ; Rabbits

Objective To construct an eukaryotic expression vector for TAP genes fused with enhanced green fluorescent protein(EGFP)gene,and to analyze the expression and subcellular localization of the fusion protein in A375 human malignant melanoma cells transfected with the eukaryotic expression vector.Methods A375 cells were cotransfected with the combination of plasmid(P)TAP1-EGFP or pTAP2-EGFP and pDsRed2-endoplasmic reticulum(ER),or with pEGFP-TAP1 and-TAP2,or monotransfected with pTAP1-EGFP or pTAP2-EGFP alone.The monoclonal A375 cells stably expressing TAP were obtained by G418 selection.Then.the distribution and expression of fusion proteins were assessed in A375 cells by using fluorescence microscopy and Western blot,respectively.Flow cytometry was used to measure the expression of HLA class Ⅰ on A375 cells.Results Transfection of A375 cells with pTAP1-EGFP or pTAP1-EGFP and pTAP2-EGFP significantly increased the expression of TAP 1 and TAP 2 in as well as HLA class Ⅰ antigen on A375 cells.The green fluorescence of TAP1-EGFP and TAP2-EGFP overlapped with the red fluorescence of ER marker in cotransfected cells.indicating that TAP was localized subcellularly on the ER.Conclusions The expression vector for TAP-EGFP fusion gene has been constructed cuccessfully and expressed in A375 cells,and the expressed fusion protein is subcelluiarly localized to ER.This study will provide a basis for the research into subsequent immune response following induction of TAP expression.

Objective To study the significance of the sample S/Co ratio when using a domestic reagent for anti-hepatitis C virus(HCV)antibody detection and to explore the procedure and standard of anti-HCV antibody diagnosis by using this domestic reagent.Methods Anti-HCV antibody was detected in 295 000 blood donors by a domestic anti-HCV reagent with enzyme-linked immunosorbent assay(ELISA)method and the reactive samples were tested again by ortho anti-HCV antibody reagent.The samples which anti-HCV antibodies were determined as positive by ortho anti-HCV rea- gent were examined by recombinant immunoblot assay(RIBA)reagent and 106 samples of them were also tested for HCV RNA.Results Six hundred and eighty-one samples were reactive in 295 000 samples screened by the domestic ELISA reagent,the reactive ratio was 0.23 %.Among the reactive samples screened by the domestic ELISA reagent,367 samples were determined as positive by ortho anti-HCV reagent while 66.2% of them showed a S/Co ratio≥3.8.The consistency rate between positive results determined by the domestic reagent and RIBA reagent respectively was 53.8%.For the samples showing S/Co ratio≥3.8 by ortho anti-HCV reagent,94.2% had a S/Co ratio≥8.0 when using the domestic ELISA reagent,while the percentage of samples showing S/Co ratio

Animals ; Brain-Derived Neurotrophic Factor ; pharmacology ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; Endothelial Cells ; cytology ; drug effects ; physiology ; Female ; Humans ; Mice ; Mice, Inbred C57BL ; Neovascularization, Physiologic ; drug effects ; Vascular Endothelial Growth Factor A ; pharmacology

This study examined the impacts of intrauterine murine cytomegalovirus (MCMV) infection on the long-term learning and memory of offspring. Sexually matured male and female BALB/C mice without MCMV infection were identified by ELISA and then mated. Seventy pregnant mice were randomly divided into the virus group (n=40) and the control group (n=30), in which the pregnant mice were subjected to placenta inoculation of MCMV suspension (1 μL, 1×106 PFU) or the same amount of cell culture medium, respectively, at gestational age of 12.5 days. Some pregnant mice [virus group (n=20), control group (n=15)] were sacrificed by cervical dislocation at gestational age of 18.5 days, and the head circumference and brain weight of the mouse fetuses were measured, and the MCMV infection in their brain tissues was detected by PCR. The other pregnant mice [virus group (n=20), control group (n=15)] delivered naturally, and the learning and memory capability of the offspring at 70-day-old was analyzed by Morris water maze test. The results showed that 28.57% mouse fetuses in the virus group developed viral infection in the brain. Their head circumference and brain weight were significantly reduced as compared with those in the control group (P<0.01). The Morris water maze test revealed that the mouse offspring in the control group found the platform with straight-line trajectories after training. In contrast, the counterparts in the virus group intended to enter the central area, but looked for the platform with a circular trajectory. And the infected mice exhibited prolonged swimming distance and swimming latency (P<0.01). It was concluded that: (1) placenta inoculation of MCMV can cause fetal brain infection and intrauterine development retardation; (2) the offspring of MCMV placenta inoculation mice showed a long-term decline in learning and memory capability.

Objective To explore the expression of transporter associated with antigen processing (TAP) and major histocompatibility complex class (MHC)-Ⅰ molecule in malignant melanoma cell lines. Methods Three malignant melanoma cell lines, including A375, A875, and KZ28 cells as well as normal melanocytes were cultured. Western blot, reverse transcription PCR and flow cytometry were used to detect the protein and mRNA expression of TAP as well as the membrane expression of MHC-Ⅰ in these cells. Results A significant decrease was observed in the expression of TAP mRNA (t = 5.89, 4.45 and 4.57 re-spectively, all P< 0.01) and protein (t= 5.46, 4.32 and 4.67 respectively, all P< 0.01) in A375, A875 and KZ28 cells compared with the melanocytes, with the strongest decrease occurring in A375 cells. Similarly, the expression of MHC-Ⅰ molecule was significantly lower in A375, A875 and KZ28 cells than that in the melanocytes (t= 6.16, 5.22 and 5.61 respectively, all P< 0.01).Conclusions The protein and mRNA expres-sion of TAP is down-regulated in three melanoma cell lines A375, A875 and AZ28, which may contribute to the escape of melanoma cells from human immune surveillance.

Objective To investigate the expression and distribution of cellular FLICE-inhibitory protein (c-FLIP) in peripheral blood and lesions of psoriatic patients. Methods Peripheral blood and skin samples were obtained from 30 patients with psoriasis vulgaris and 20 normal controls. Flow cytometry was used to detect intracellular c-FLIP protein in peripheral T and B lymphocytes, immunohistochemistry to examine the expression of c-FLIP in lesional tissue. Results Based on the positivity rate of c-FLIP, there was a significant increase in T lymphocytes in active psoriasis compared with regressive psoriasis and normal controls (6.32%±1.17% vs 2.64%±0.74% and 2.28%±0.54%, P＜0.01 and 0.05, respectively), while no significant difference was found in B lymphocytes among these three groups (0.78%±0.16%, 0.71%±0.32%, 0.69%±0.18%, respectively, P＞0.05). The expression intensity of c-FLIP in keratinocytes was also higher in active psoriasis than in regressive psoriasis and normal controls (89.73±5.24 vs 117.40±7.50,121.58±7.93, P＜0.01 and 0.05 respectively), and there was no difference between regressive psoriasis and normal controls (P＞0.05). Conclusions c-FLIP is highly expressed in lesions and peripheral T lymphocytes of patients with active psoriasis, suggesting the possible involvement of c-FLIP in the proliferation of T lymphocytes in psoriasis.

Objective To study the expression of cellular FLICE inhibitory proteins(c-FLIP)in peripheral blood B lymphocytes in patients with systemic lupus erythematosus (SLE)and its correlation with clinical features.Methods Blood samples were obtained from 53 patients with SLE and 30 normal human controls.Flow cytometry and ELISA were performed to measure the expression of c-FLIP in pefipheral blood B lymphocytes and serum levels of IL-4 and IL-10,respectively.Relevant laboratory examinations were carried out for these patients.SLE disease activity index(SLEDAI)score was calculated for patients.Results The positivity rate of c-FLIP in B lymphocytes was 3.11%±0.70%in 18 patients with active SLE.significantly higher than that in 35 patients with inactive SLE (0.78%±0.28%)and normaI controls(0.68%±0.12%),while no statistical difierence was found between inactive patients and controls(t=1.56,P＞0.05).In SLE patients,the expression of c-FLIP showed a positive correlation with SLEDAl score(r=0.96.P＜0.05),erythrocyte sedimentation rate(r=0.96,P＜0.01),serum level of C reactive protein(r=0.92.P＜0.01)and the titer of antinuclear antibodies(r=0.86,P＜0.01),whereas in 36 patients with leucopenia.a negative correlation was noticed between white blood cell count and the expression level of c-FLIP(r=-0.94,P＜0.0 1).The 23 patients with lupus nephritis had a higher level of c-FLIP than those without lupus nephritis(3.04%±1.09%vs 1.76%±1.09%,t=4.23,P＜0.05).Additionally.the expression of c-FLIP positively correlated with the serum level of IL-4 and IL-10(r=0.80,0.89.respectively,both P＜0.01).Conclusions In patients with active SLE,the expression of c-FLIP is upregulated in peripheral blood B lymphocytes,and positively correlated with the severity of SLE as well as the serum level of IL-4 and IL-10.The upregulation of c-FLIP in B cells might play a certain role in deficient apoptosis or clearance of activated B cells in SLE.