Objective To investigate the phenotype and function of antigen-specific CD8+ cytotoxic T cells (CTL) from HLA-A2+ healthy human donors induced by high-carcinogenic HPV 16 E7 peptide in vitro.Methods Peripheral blood T cells from 26 HLA-A2+ healthy human donors were incubated with HPV 16E711-20 peptide (HLA-A*0201/YMLDLQPETT), recombinant human interleukin 7 (IL-7) and IL-2 for 7 days to yield antigen-specific CD8+ CTL. Then, four-color flow cytometric analysis was performed to detect the percentage of antigen-specific CD8+ CTL expressing different surface markers including CD45 and CD27. The expression of three intracellular cytokines including perforin, granzyme-B and FasL in the antigen-specific CD8+ CTL was also measured by intracellular flow cytometry. A wrong peptide, HBVcoxe18-27 (FLPSDFFPSV),was used as an isotype control. Results A significant increase was observed in the percentage of antigen-specific CD8+ CTL in peripheral T cells stimulated with HPV 16 E7-peptide compared with the non-stimulated T cells (0.73% ± 0.33% vs 0.02% ± 0.03%, P ＜ 0.01). The percentage of CD45RA+CD27- effector T cells,CD45RA-CD27- effector memory T (TEM) cells, CD45RA-CD27+ central memory T (TCM) cells, and CD45RA+CD27+ naive T cells in antigen-specific CD8+ CTL was 26.07% ± 13.46%, 7.97% ± 7.11%, 33.25% ± 19.68%and 32.73% ± 13.89%, respectively in HPV 16 E7-stimulated group, significantly higher than that in nonstimulated group (0.02% ± 0.03%, 0.02% ± 0.03%, 0.02% ± 0.03% and 0.02% ± 0.05%, all P ＜ 0.01 ). Elevated proportions of perforin-, granzyme-B- and FasL-expressing antigen specific CTL were observed in HPV 16 E7-stimulated group compared with non-stimulated group (47.01% ± 18.69% vs 0.38% ± 0.55%, 80.53% ±13.32% vs 0.34% ± 0.22%, 26.48% ± 7.81% vs 0.16% ± 0.16%, all P ＜ 0.01 ). Conclusions HPV 16 E7peptide could induce the clonal proliferation of CD8+ T cells, generation of antigen-specific CD8+ CTL and secretion of toxic cytokines, finally lead to a highly efficient and specific killing of virus-infected target cells through different mechanisms, hence, it might play a crucial role in antiviral immune responses.

Technologies for the trans-institutional sharing of scientific data were studied with the sharing of nucleic acid sequence data as the study object , such as leaving a flexible space for implementation of the unified standards in working out the criteria for data sharing , implementing regular point to point data sharing and updating in agree-ment of institutional league , and providing multiple data services according to the unified working process by giving considerations to the local demands .The significance of trans-institutional sharing of scientific data under the insti-tutional league model was summarized for the reference in constructing the scientific datasharing platform.

Objective To investigate the expression of activating receptors (NKG2D and NKp46), inhibitory receptors (NKG2A and KIR) as well as costimulatory molecules (OX40, 4-1BB and ICOS) on peripheral blood natural killer (NK) cells from patients with recurrent genital herpes (RGH). Methods Four-color immunofluorescence staining with flow cytometry was used to detect the expression of NKG2D, NKG2A, KIR and NKp46 in 44 patients with RGH and 40 normal human controls, and to detect the expression of OX40, 4-1BB and ICOS in 29 patients with RGH and 29 normal human controls. Results The proportions of NKG2D-positive and NKp46-positive NK cells significantly decreased in patients with RGH than those in the normal human controls [(93.3 ± 5.4)% vs (96.9 ± 2.5)%, (88.9 ± 8.7)% vs(93.4 ± 4.1)%, respectively, both P < 0.01]. Between the patients and controls, no significant difference was observed in the expression of NK cell inhibitory receptors, NKG2A [(41.8 ± 14.4)% vs (46.0 ± 14.7)%, P > 0.05] or KIR [(68.3 ± 19.1)% vs (69.1 ± 17.6)%, P > 0.05]. A lower expression of costimulatory molecule OX40 was noted in NK cells from patients with RGH compared with those in normal controls [(1.0 ± 1.1)% vs (1.8 ± 1.7)%, P < 0.05]. Conclusions Herpes simplex virus infection could down-regulate the expression of NK cell activating receptors and costimulatory molecules, subsequently suppress the activation of NK cells, and lead to the escape of virus-infected cells from the killing of NK cells.

Objective To explore the expression of GATA-3 in pulmonary tissue of asthmatic mice and the inhibitory effect of dexamethsone(Dex)on it.Methods The Blab/c mice asthma model was induced by ovalbumin(OVA) with classic method.Twenty-four male mice were randomly divided into control group,asthmatic group and Dex treated group.The expression of GATA-3 protein was measured by immunohistochemical staining.The expression of GATA-3 mRNA was measured by reverse transcription-polymerase chain reaction(RT-PCR).The level of IL-4 in mice spleen CD4 T cell was measured by flow cytometry.The airway inflammation was evaluated by HE staining.Results The percentages of postive GATA-3,GATA-3 mRNA and IL-4 protein of asthmatic group were significantly higher than those of control group(P

Objective To observe the ultrasonographic features of local chest wall tumor recurrence after mastectomy for breast cancer and its clinical and histopathological characteristics. Methods The ultrasonographic features, clinical and histopathological characteristics of 27 patients with local chest wall tumor recurrence after mastectomy confirmed histopathologically were retrospectively reviewed. Results The disease-free intervals of twenty-seven patients ranged from 3 to 129 months [mean (31.9±31.4) months]. Most of the recurrence(18/27, 66.7%) occurred within 3 years after mastectomy. The clinical manifestations were:7 cases (7/27, 25.9%) with regional skin redness and swelling, red rash or ulceration on chest wall associated with or without palpable mass, 20 cases(20/27, 74.1%) with chest wall palpable masses without obvious skin change. On ultrasonography, 2 cases showed diffuse inifltrative type with ill-deifned inhomogeneous hypoechoic lesion and skin thickening. And twenty-ifve cases(43 lesions) showed mass type with a lesion size range of 5.4-114.7 mm [mean (24.4±21.6) mm]. Among them, 32 lesions were located near to the operation incision scar, 36 involved muscle layer, 38 were hypoechoic, 31 had irregular shape, 24 had indistinct margin, and 31 had blood lfow signal. In addition, calciifcation, halo, and taller-than-wide shape were absent in all 43 lesions. Conclusions The tumor recurrence often occurred within 3 years after mastectomy in high-risk patients. Ultrasonographic feature of chest wall recurrent lesion is of great value in the diagnosis.

In order to investigate the role of Toll-like receptor 4 (TLR4) in cerebrocardiac syndrome (CCS), the partial cerebral ischemia/reperfusion (I/R) models in mice with different TLR4 genotypes were established in the present study. TLR4 wild-type (C3H/HeN) and mutant (C3H/HeJ) mice of 6-8 weeks of age were divided into 4 groups at random: C3H/HeN sham group (n=10), C3H/HeJ sham group (n=10), C3H/HeN model group (n=10) and C3H/HeJ model group (n=10). Partial cerebral I/R was caused by the middle cerebral artery occlusion (MCAO) to duplicate CCS models in mice. After the operation, the electrocardiogram (ECG), the level of tumor necrosis factor-alpha (TNF-α) in myocardial tissue and the cardiac pathological changes were observed in each group. It was shown that the brain infarct volume in C3H/HeN model group was larger than that in C3H/HeJ model group (P<0.01). The ST segment change and T wave inversion occurred frequently in model groups. Moreover, the TNF-α level in C3H/HeN model group was higher than that in C3H/HeJ model group (P<0.01). The myocardial injury was aggravated in C3H/HeN group as compared with C3H/HeJ group. It was concluded that TLR4 was implicated in the development of CCS.

Objective To explore the psychomotor development in 12 month old infants born after in vitro fertilization with a control group of infants concieved naturally.Methods A matched control study was performed on psychomotor development of 12-month-old infants concieved with use of assisted reproductive technology(in vitro fertilization only).The control group was matched according to maternal age,parity,social class and level of parental education.The suitable mothers were invited to participate at the 28th week of gestation and were followed up to delivery.The infants of the two groups were followed up to 12 months old and a formal developmental assessment was done with the Children Development Center of China(CDCC) scales of infant development.Results The incidences of preterm birth,low birth weight and twin pregnancy were significantly higher in the assisted conception group.No statistically significant difference was found in the mental development index and psychomotor development index between assisted conception and control groups.Conclusions The levels of psychomotor development in 12 months old infants born after in vitro fertilization are normal.But as the incidences of preterm birth,low birth weight and multifetation are significantly higher in vitro fertilization group,it need to follow up the ongoing development of these children.

Objective To study the expression of S100A8 and the relationship between S100A8 and Toll-like receptor 4 (TLR4) in focal cerebral ischemia reperfusion (I/R) injury.Methods C3H/HeJ mice with TLR4 gene mutation (n =30) and C3H/HeN with normal TLR4 gene mice (n =30) were divided into 4 groups at random (random number),namely C3H/HeJ model group (n =18),C3H/HeJ control group (n =12),and C3H/HeN model group (n =18).C3H/HeN control group (n =12).Middle cerebral artery was occluded to make I/R model in mice by using thread embolism method.Brain tissues were collected after ischemia for one hour and reperfusion for 12 hours.Stroke outcome was evaluated by determination of infarct volume of brain tissue and assessment of neurological scores.And brain injury after cerebral I/R was observed by optical microscope after TTC and HE staining.The immunofluorescence technique and RT-PCR were used to determine the protein level and expression of S100A8 mRNA in damaged brain tissues.Results Compared with C3H/HeN model mice,TLR4-deficient model mice (C3H/ He J) had lower infarct volumes and better outcomes of neurological function after resuscitation for 12 hours.Compared with control groups,the expression of S100A8 mRNA and level of S100A8 protein increased greatly in damaged brain tissues of model mice after I/R injury.In addition,model mice with lacked TLR4 (C3H/HeJ) had lower expression of I/R-induced S100A8 mRNA than C3H/HeN mice in model group,indicating that the close relationship between the levels of S100A8 and TLR4.Conclusions S100A8 interaction with TLR4 might be involved in brain damage and in inflammation triggered by I/R injury.

OBJECTIVETo observe the effect of β₃adrenoceptors (β₃-AR) activation on rat thoracic aorta smooth muscle contractility and the possible related mechanism.

METHODSThe endothelium removed thoracic aorta was pre-contracted with 30 mmol/L KCl physiological saline solution (PSS). Then the tension of the thoracic aorta was recorded in presence of BRL37344 (BRL) to determine the action of β₃-AR. The tension of the thoracic aorta was also recorded in the presence of Propranolol (PRA), SR59230A (SR), L-NNA, H-89 and Iberiotoxin (IBTX) respectively to reveal the underling mechanism of β₃-AR activation on rat vascular smooth muscle. Immunohistochemistry was adopted to confirm the existence and the distribution of β₃-AR in rat thoracic aorta.

RESULTSThe results showed that: (1) The thoracic aorta was relaxed by β₃-AR activation, with a relaxation percentage of (10.59 ± 0.79). (2) β₃-AR was expressed in both endothelial and smooth muscle layer in thoracic aorta sections of rats. (3) PRA did not block the effect of BRL on the thoracic aorta. The relaxation actions of BRL could be antagonized by pre-incubating the thoracic aorta with SR. (4) L-NNA (a NOS inhibitor) and H-89 (a PKA inhibitor) reversed the relaxation effect of BRL on vascular smooth muscle. (5) The effect of BRL was decreased after application of Ibriotoxin (IBTX), a large conductance calcium dependent potassium channel blocker.

CONCLUSIONThe results confirmed that activation of β₃-AR led to relaxation of thoracic aorta smooth muscle. The relaxation action of β₃-AR on smooth muscle of rat thoracic aorta was related to activation of NOS and PKA signaling pathway. Large conductance Ca²⁺-K⁺ channels were involved in the relaxation action of β₃-AR activation on rat thoracic aorta smooth muscle.

Animals ; Aorta, Thoracic ; physiology ; In Vitro Techniques ; Isoquinolines ; Large-Conductance Calcium-Activated Potassium Channels ; physiology ; Muscle Contraction ; Muscle Relaxation ; Muscle, Smooth, Vascular ; physiology ; Nitroarginine ; Peptides ; Propanolamines ; Propranolol ; Rats ; Receptors, Adrenergic, beta-3 ; physiology ; Signal Transduction ; Sulfonamides

Animals ; Biogenic Polyamines ; metabolism ; Biological Transport ; Humans ; Paraquat ; poisoning ; Pulmonary Edema ; metabolism