Objective:To prepare and test tetrameric sH-2K~d-HBc complex for the further measurement of the specific CTL response.Methods:PE labled streptavidin with 4 biotinylated binding sites can bind to 4 biotinylated monomer to form the corresponding tetramer.Mice were immunized via different methods of genetic immunization by use of the construted pcDNA3-C plasmid to get the specific CTLs.Then our prepared tetramer was applied to stain the specific CTLs by the analysis of flow cytometry.Results:We applied our prepared tetramer to stain the cells from the experimental groups and control group.The results showed the tetramer was able to discriminate the frequencies of specific CTL induced by the three immunol methods(0.24%,0.26%,0.36% vs 0.07%,P≤0.05).This demonstrated that the prepared tetramer could bind its targets specifically and efficiently.The three immunol methods induced different levels of immune responses.Compared with the traditional muscle injection,gene gun induced weaker humoral immune response and stronger cellular immune response,and hydrodynamic injection induced the strongest humoral and cellular immune responses.Conclusion:Have successfully constructed the sH-2K~d-HBc tetramer.The techniques and methods can be used for preparation of tetramers of other types of MHCⅠ molecules.

Objective:To clarify the differences of host immune responses at different stages of HBV infection. Methods:We constructed three HLA-A*0201/HBV tetramers with immunodominant epitopes of core18-27,polymerase 575-583 and envelope 335-343,and analyzed antigen specific CD8+ T cells and the expression of CD127 in peripheral blood mononuclear cells ( PBMCs) from patients infected with HBV using these HLA-A*0201/HBV tetramers. Results: The frequencies and expansion ability of antigen specific CD8+ T cells in most self-limited HBV infected individuals were higher than that in chronically HBV infected patients. In low copy period the frequencies of antigen specific CD8+ T cells were similar to those in immune clearance phase at a high viral load and liver damage and in immune clearance phase, which had no significant correlation with virus quantitation and ALT level. In chronic infection the ability of antigen specific CD8+ T cells proliferation was inversely proportional to the viral titer. In most self-limited HBV infected individuals the IFN-γsecretion functions of antigen specific CD8+ T cells were higher than in chronic infection,but in immune tolerance phase these cells lost the ability. HBsAg level was different at different stages after HBV infection:it was highest in immune tolerance phase,but in immune clearance phase,activity period and low copy period the correlation with HBV DNA replication gradually declined. The frequency of CD8+ CD127+ T cells in chronic HBV infection was lower than the control group and self-limited infection group,especially in immune tolerance with HBeAg+ and immune clearance phase. Conclusion: The frequencies of antigen specific CD8+ T cells are not the main determinant of immune-mediated protection in chronic HBV infection,memory antigen specific CD8+ T cells are not clear or missing,which provides the possibility for therapeutic vaccines and immunization therapy.

BACKGROUND: Previous studies have confirmed that 6-hydroxydopamine is capable to increase the expression of divalent metal transporter-1 and reduce the expression of ferroportin-1 in the neurons and microglia, which may lead to iron deposition in the substantia nigra after Parkinson’s disease. However, it is unclear whether 6-hydroxydopamine can play diverse roles in astrocytes.
OBJECTIVE:To observe the effects of 6-hydroxydopamine on the expression of divalent metal transporter-1 and ferroportin-1 in rat C6 glioma cel lines.
METHODS:C6 glioma cell lines from rats were cultured in 10 μmol/L 6-hydroxydopamine for 24 hours. Then, protein expressions of divalent metal transporter-1 and ferroportiner-1 were measured by western blot method.
RESULTS AND CONCLUSION:The protein expressions of divalent metal transporter-1 and ferroportin-1 in C6 glioma cell lines were increased by 2.5 times (P < 0.01) and 1 time (P < 0.05), respectively, after treatment with 6-hydroxydopamine. These findings indicate that 6-hydroxydopamine can promote iron transport rate in astrocytes by increasing both divalent metal transporter-1 and ferroportin-1 expressions, and astrocytes has a different response to 6-hydroxydopamine from neurons and microglia.

Hepatitis B virus (HBV)-specific cytotoxic T lymphocytes (CTLs) are believed to play a major role in viral clearance and disease pathogenesis during HBV infection. To clarify the differences in host immune responses between self-limited and chronic HBV infections, we constructed three HLA-A*0201/HBV tetramers with immunodominant epitopes of core18-27, polymerase 575-583 and envelope 335-343, and analyzed the HBV-specific CTLs in peripheral blood mononuclear cells (PBMCs) from patients infected with HBV. The frequencies and expansion ability of HBV-specific CD8(+) T cells in most self-limited HBV infected individuals were higher than those in chronic HBV-infected patients. HBV-specific CD8(+) T cells could be induced by in vitro peptide stimulation from chronic patients with a low level of serum HBV-DNA but not from those with a high level of serum HBV-DNA. In chronic infection, no significant correlation was found either between the frequencies of HBV-specific CD8(+) T cells and the viral load, or between the frequencies and the levels of alanine transaminase. Our results suggested that the frequencies of HBV-specific CTLs are not the main determinant of immune-mediated protection in chronic HBV infection and immunotherapeutic approaches should be aimed at not only boosting a HBV-specific CD8(+) T response but also improving its function.

Previous studies have shown that expression of the interferon-sensitive gene (ISG)I5 protease UBP43 is increased in the liver biopsy specimens of patients who do not respond to interferon (IFN)-α therapy. We hypothesized that UBP43 might hinder the ability of IFN to inhibit HBV replication. In this study, we investigated whether vector-based siRNA promoted by Hi (psiUBP43) could enhance IFN inhibiting HBV replication in cell culture. UBP43 was specifically silenced using shRNA. In HepG2.2.15 cells, the HBeAg and HBV DNA levels were significantly reduced by IFN after transfection of shRNA, imphicated that vector-based siRNA promoted by HI (psiUBP43) could enhance IFN inhibiting HBV replication in cell culture. These data suggest that UBP43 modulates the anti-HBV type I IFN response, and is a possible therapeutic target for the treatment of HBV infection.

Objective To evaluate biochemical characteristics and the trend of diabetic complications in patients with newly-diagnosed type 2 diabetes from 1994 to 2008. Methods We utilized the database of the diabetes complications assessment and analyzed the metabolic disorder and the diabetic complications in the patients with newly diagnosed diabetes. Results 2 085 cases were collected, including 1189 males and 896 females. The average age of onset of diabetes was 51.6±13.1 and 54.6±7.9 yrs respectively in 2008 and 1994. During 1994,no case was found in subjects aged 20-29 yrs and 5% of the patients were aged 30-39; but 2% of patients aged 20-29 and 16% aged 30-39 yrs were found in 2008. BMI was increased from 24.48±4.15 in 1994 to 26.03±3.63 in 2008. Percentage of patients with abnormal BMI ( ≥25 kg/m2 ), WHR [≥0.90 (male) or ≥0.85 (female)]increased significantly from 63.6%, 75.0%, and 71.4% in 1994 to 79.6%, 95.2%, and 93.8% in 2008,respectively. Both SBP and DBP were not significantly changed. The fasting blood and postprandial blood glucose,HbA1c decreased from 10.3 mmol/L, 15.2 mmol/L, 11.1% in 1994 to 9.0 mmol/L, 14.3 mmol/L, and 8.6% in 2008, respectively. The average TG level increased from 1.7 mmol/L in 1994 to 2. 1 mmol/L in 2008,however, TC and HDL level were not significantly changed. The prevalence of diabetic retinopathy decreased from 28.2% in 1994 to 3.9% in 2008. The prevalence of diabetic nephropathy increased from 17.7% in 1994 to 24.6% in 2008. The prevalence of diabetic cardiovascular disease increased from 14.3% in 1994 to 24. 1% in 2008. Compared with the patients without microvascular complications, the patients with microvascular complications had higher SBP, DBP, and HbA1c( 136/78 vs 130/77 mm Hg, 9.41% vs 9.11% ). The patients with macrovascular complications had older age, higher SBP, TC, and TG than those without macrovascular complications (53.4 vs 50.0 yrs; 132 vs 129 mm Hg ; 5.3 vs 5.1 mmol/L and 2.6 vs 2.1 mmol/L). Conclusions In the studied newly-diagnosed diabetic patients from 1994 to 2008, there were increasing incidences of obesity and hypertriglyceridemia. However, the prevalence of diabetic retinopathy decreased significantly, while that of nephropathy showed no significant change.Cardiovascular complications were markedly increased.

OBJECTIVE: To explore the found ways and first diagnosis age of children with large vestibular aqueduct, and their relations with hearing loss. METHOD: Medical histories of 122 cases of children diagnosed with large vestibular aqueduct by HRCT or MRI had been collected from January 2009 to April 2014 in our hospital children's hearing diagnosis center clinic. Found ways comprise of accepting universal newborn hearing screening (UNHS) group and unaccepting UNHS group. Accepting UNHS children were divided into two ears unpassing group, single ear unpassing group and passing group. The patients in unaccepting UNHS group were divided into not sensitive to sounds, speech stunting, sudden hearing loss, and other group. Analysis the relationship between the found ways and first diagnosis age and their relations with hearing loss. RESULT: There are 84 cases (68.85%) accepting UNHS, the average age of first diagnosis was (17.24 ± 17.08) months; 37 cases (31.15%) are not accepting UNHS. The average age of first diagnosis was (30.92 ± 18.21) months. The average first diagnosis age of accepting UNHS group was more earlier than the unaccepting UNHS group. The difference was statistically signif- icant (P < 0.01). There were 57 cases (67.85%) whose two ears not pass UNHS; 15 cases (17.86%) single ear not pass; namely the referral rate was 85.71%; 12 cases (14.29%) pass the test. The first diagnosis age of passing UNHS group was more later than two ears unpassing group (P < 0.001). In the unaccepting UNHS group, the average first diagnosis age of not sensitive to sounds group (19.69 ± 11.16 months) was more earlier than words dysplasia group (37.13 ± 15.62 months) and sudden hearing loss group (47.40 ± 24.70 months) (P < 0.01). The difference in the degree of hearing loss between accepting UNHS and unaccepting UNHS group had no statistical significance (P > 0.05). In unaccepting UNHS group ,the average first diagnosis age of the mild-to-moderate hearing loss group was later than the very severe hearing loss group (P < 0.01). CONCLUSION Most of large vestibular aqueduct children can be found and receive diagnosis early by UNHS. But part of these patients with late-onset or progressive hearing loss, especially these with mild-to-moderate hearing loss cannot be found early, which should arouse our attention.
Child, Preschool ; Deafness ; Early Diagnosis ; Hearing Loss ; etiology ; Hearing Loss, Sudden ; Hearing Tests ; Humans ; Infant ; Infant, Newborn ; Neonatal Screening ; Vestibular Aqueduct ; abnormalities

Activity of IL-1 produced by peripheral blood monocytes stimulated with lipopolysaccharide in vitro was detected from 17 acute cases, 14 chronic cases and 19 advanced cases of schistosomiasis japonica. It was found that the level of IL-1 was significantly increased and positively related to the body tempereture in the group with acute schistosomiasis. The activity of IL-1 was statistically reduced in the chronic and advanced groups, especially in the latter. After inhibiting the synthesis of prostaglandin with indomethacin, the level of IL-1 was significantly increased in three groups of patients, but no apparent change in the normal contro group. The results indicate that IL-1 may play an important role in inducing the inflammatory reaction in patients with acute schistosomiasis japonica and in the immunoregulation in the chronic stage. The changes of IL-1 activity in patients with schistosomiasis japonica may be closely related to prostaglandin.

Early mobilization plays a vital role in the recovery of patients after lung cancer surgery. From the perspective of enhanced recovery after surgery, this paper summarized the mobilization program, effect evaluation and expanded application of early getting out of bed in lung cancer patients, in order to provide reference for medical staff to formulate early mobilization programs.

Objective Explore an approach with which stem cell can be used to cure traumatic spinal cord. Methods To better repair spinal cord injury, adult Sprague-Dawley (SD) rat MSCs was isolated and induced to differentiate into neuron-like cells with a Chinese medicine, Musk's polypeptide named Musk-1 in vitro and be seeded into microsurgical transectional model of adult rat spinal cord with cell differentiation and transplantation techniques. Results After cell transplantation, animals seeded with the rMSCs-derived neuron-like cells promoted significant improvement in function after spinal cord (P＜0.05,effective observation for 90 days ) when compared to the lesion-control group. Histology and immunocytochemical analysis confirmed a large number of rMSCs-derived neuron-like cells survived well in the transplantation zoon and numerous cells migrated within the host adjacent tissues as far as 6millimeters above and below the border of the lesion. Fluorescence gold retrograde tract tracing analysis revealed the positive motor neuron of fluorescence gold mark was found in the head side of rat spinal marrow ,corpus rubrum ofmesencephalic and corticospinal tract fibers. This suggested that corticospinal tract fibers of spinal cord area got regeneration and passed through the lession to reach the tail of spinal marrow.Conclusion These findings demonstrated that "pre-programmed" rMSCs can survive, migrate, integrate as well as restore spinal function and behavior in the microsurgical transectional model of rat spinal cord, and may serve as a suitable approach to traumatic spinal cord injury.