Objective To explore the effect of DR on the expression of inhibitory neurotransmitter ,γ-aminobutyric acid ( GABA) and brain derived neurotrophic factor ( BDNF) in the primary visual cortex of young cats .Methods Totally 6 cats were wsed in this study . Nissl staining was used for cortical layer identification and cell counting . Immunohistochemical techniques were utilized to label GABA-and BDNF-positive neurons .Sections were observed under an Olympus light microscope and photographed with a digital camera .The cell density and absorbance of immunoreactivity were measured with Image-Pro Express 6.0 softwares .Results Our results showed that the mean density of Nissl-stained neurons in the primary visual cortex ( V1) of the DR group showed no significant difference from that of the control group . However , the mean density of GABA-immunoreactive neurons in each cortical layer of V 1 in DR cats was significantly higher than that in control cats .The mean immunoreactive intensity of GABA-positive neurons , as indicated by the average absorbance , increased significantly in DR cats relative to control ones . Accompanied with the elevation of GABA expression, DR enhanced BDNF expression in V1, as indicated by an increased mean density of BDNF-positive neurons and BDNF-immunoreactive average absorbance in DR cats relative to controls .Conclusion These results indicate that dietary restriction leads to a concurrent expression upregulation of GABA and BDNF , which may likely compensate for brain functional degradation during senescence and thus delay aging process .

Objective:The immunogenicity of DNA vaccine immunogenicity can be improved by fusing antigenic genes to IgG Fc fragment.It is critical for enhancing immunogenicity of DNA vaccine to construct the secreting eukaryotic expressing vector.The purpose of this study is to construct a secreting eukaryotic expressing vector ligated with IgG Fc gene by amplifying human IgG Fc fragment and fusing the fragment with human CD5 signal peptide sequence for highly efficient expression.Methods:Human lymphocytes were isolated from the tonsil obtained by removal surgery.The total RNA of lymphocytes was extracted using Trizol reagent.Human IgG Fc cDNA was amplified by RT-PCR from lymphocytes and then inserted into pMD-18T vector.The Fc encoding sequence fused with CD5 signal peptide(sp) sequence was constructed by cross PCR using Fc fragment and CD5sp sequence and cloned in pcDNA-CD5 plasmid as the template.CD5sp-Fc fragment was inserted into pcDNA3.1 expressing vector to construct the pcDNA-CD5sp-Fc plasmid.The plasmids were transfected into HEK293T cells mediated by calcium phosphate.Western blot was used to detect expressed Fc protein in cultural supermatant of the cells.Results:The amplified sequence of human IgG Fc was consistent with that previously published.The secretory expression of Fc in HEK293T cells was achieved and the expressing level reached 50 ?g/106 cells at 48 h culture after transfection.Conclusion:The human IgG Fc gene is amplified by RT-PCR methods and the secretory expression of Fc gene mediated by CD5 signal peptide in 293T cells is achieved.The results provide a experimental basis for further construction of antigen genes fused to IgG Fc fragment and investigation on DNA vaccines and Fc as a biological adjuvant.

Objective To investigate the short-term efficacy of CT-guided radiofrequency ablation (RFA) for the treatment of advanced non-small cell lung cancer. Methods During the period from June 2010 to June 2013, a total of 100 patients with advanced non-small cell lung cancer were admitted to authors’ hospital. The patients were equally and randomly divided into the study group (n=50) and the control group (n=50). The patients of the control group received concurrent radiotherapy and chemotherapy treatment, while the patients of the study group received CT-guided RFA. The clinical effect, changes in CT values after the treatment, the improvement of physical condition and postoperative complications were recorded, and the results were compared between the two groups. Results The remission rate of the disease in the study group was 86.0%, which was much higher than that in the control group (52.0%), and the difference between the two groups was statistically significant (P<0.05). After the treatment, the CT value of the study group was (14.1±3.9) HU, which was significantly lower than that of the control group (29.8±4.7 HU, P<0.05). The physical improvement rate of the study group (66.0%) was significantly higher than that of the control group (44.0%), the difference between the two groups was statistically significant (P<0.05). The occurrence of postoperative complications, such as pulmonary infection, chest pain, fever, thoracic effusion and gastrointestinal reaction, in the study group was strikingly lower than that in the control group (P<0.05). Conclusion For the treatment of advanced non-small cell lung cancer, CT-guided radiofrequency ablation is safe and less-invasive, it can improve the clinical short-term effect as well as the quality of life. Therefore, this technique should be recommended in clinical practice.

Objective:To investigate the quantification of CD4~+CD25~+ regulatory T cells and distribution of CD4~+CD8~+ T lymphocyte subgroup in peripheral blood of patients in chronic hepatitis B (CHB),and to reveal relationship between CD4~+CD25~+ regulatory T cells,CD4~+CD8~+ T lymphocyte subgroup and HBV infetion as well.Methods:CD4~+CD25~(high),CD4~+CD25~+Foxp3~+Treg and CD3~+CD4~+CD8~+T lymphocyte subgroup in peripheral blood from 50 patients with CHB and 20 healthy controls was analyzed using flow cytometry.HBV DNA was detected by fluorescence quantitative PCR.Results:The number of CD4~+CD25~(high)Tregs in patients with CHB was obviously higher than that in healthy controls(P＜0.01)and increased with copies of HBV DNA.The same with the change of CD4~+CD25~+Foxp3~+Tregs in patients with CHB and there was a positive correlation between CD4~+CD25~(high)Tregs and CD4~+CD25~+Foxp3~+Tregs(r=0.890,P＜0.001).Compared with healthy controls,the frequency of CD4~+T cells and the ratio of CD4~+/CD8~+ in patients with CHB was declined,but there was no significant difference in the frequency of CD3~+T cells and CD8~+T cells between them(P＞0.05).The variation in the number of CD4~+CD25~(high)Tregs was correlated positively with the copies of HBV DNA(r=0.782,P＜0.001)and glutamic-pyruvic transaminase(ALT)(r=0.432,P＜0.005)separately,but negatively with the frequency of CD3~+,CD4~+,CD8~+T cells and the ratio of CD4~+/CD8~+(P＞0.05).The variation in the frequency of CD3~+,CD4~+,CD8~+T cells and the ratio of CD4~+/CD8~+ was also correlated negatively with the copies of HBV DNA(P＞0.05).Conclusion:The number of CD4~+CD25~(high)Tregs increases in patients with CHB and is in accordance with the copies of HBV DNA and increased level of ALT.Further studies should be done to investigate weather CD4~+CD8~+ T lymphocyte subgroup could be used to monitor the state of community.

BACKGROUND: There are fewer reports about estrogen effects on bone marrow stromal cells (BMSCs). OBJECTIVE: To study the effect of diethylstilbestrol on the osteogenic differentiation of rabbit BMSCs. METHODS: Rabbit BMSCs cultured in vitro were intervened with 0, 10-7, 10-6, 10-5 mol/L diethylstilbestrol, and BMSCs cultured with dexamethasone 10-8 mol/L, β-sodium glycerophosphate 10 mmol/L, and vitamin C 50 mg/L were used as positive controls. RESULTS AND CONCLUSION: 10-6 mol/L diethylstilbestrol significantly improved the proliferative ability of BMSCs at 24, 48, and 72 hours after intervention (P < 0.01). 10-5 mol/L diethylstilbestrol significantly inhibited the proliferation of BMSCs at 48 hours after intervention as well as 10-7 mol/L diethylstilbestrol at 72 hours (P < 0.01). Mineralized nodular structures formed at 25 days after intervention with 10-7 mol/L diethylstilbestrol. Alkaline phosphatase activities were remarkably increased at 14 and 21 days after intervention with 10-7, 10-6 mol/L diethylstilbestrol. It has been proved that diethylstilbestrol has an enhancing effect on the osteogenic differentiation of rabbit BMSCs.

BACKGROUNDKAI1 is a new identified metastasis-suppressor gene whose expression in many types of tumors has been reported. The aim of study is to investigate the role of KAI1 protein in development of lung cancer and its values in predicting the prognosis of lung cancer.

METHODSThe expressions of KAI1 protein were detected in benign pulmonary disease tissue, precancerous disease tissue, lung cancer tissue and metastatic lung cancer tissue in local lymph node using tissue microarray and immunohistochemical method. The relationship between expression of KAI1 protein and clinicopathological parameters of patients with lung cancer was analyzed by Chi-Square test and Fisher exact test.

RESULTSThe positive rate of KAI1 expression was 100.0% in 10 cases of benign pulmonary diseases, 66.7% in 12 cases of precancerous diseases, 24.7% in 89 cases of primary lung cancer and 0 in metastatic lung cancer tissue in local lymph node respectively. The KAI1 protein expression in primary lung cancer tissues had no remarkable relationship with age and gender of the patients and the location of cancer, but had significant relationship with the histological type and differentiated degree of tumor, P-TNM stages and lymph node metastatic status.

CONCLUSIONSThe abnormal expression of KAI1 protein may participate in malignant progression of lung cancer. Its downregulation may promote the invasion and metastasis of tumor cell. Detection of the expression of KAI1 protein may be helpful to predict the prognosis of lung cancer.

In order to investigate the effects of phenylalanine, tyrosine and tyramine on the growth of Lycoris radiata suspension cells and the accumulation of alkaloids, the growth quantity of the cells as well as the content of alkaloids in cells were determined, which were treated with above three kinds of precursors alone and phenylalanine combined with tyrosine respectively. The results indicate that the addition of phenylalanine alone and addition of phenylalanine on the basis of tyrosine at high concentration (200 micromol/L) had no significant effect on the growth of Lycoris radiata suspension cells and the content of alkaloids in cells; whereas tyrosine and tyramine promoted the growth of the cells and alkaloids accumulation. Treated with tyrosine at high concentration (200 micromol/L), the content of alkaloids of the cells was 2.56-fold higher than that of the control group, the amounts of lycoramine (3.77 mg/g) and galanthamine (4.46 mg/g) were 6.61-fold and 6.97-fold higher than that of the control group, respectively. When treated with tyramine (200 micromol/L), the amount of alkaloids in Lycoris radiata suspension cells was 2.63-fold higher than that of the control group, and the amounts of lycoramine (4.45 mg/g) and galanthamine (5.14 mg/g) were 9.08-fold and 9.18-fold higher than that of the control group, respectively. The above results demonstrate that adding tyrosine and tyramine in the media significantly promoted the growth of the Lycoris radiata suspension cells and alkaloids accumulation in the cells.
Amaryllidaceae Alkaloids ; chemistry ; Cells, Cultured ; Culture Media ; chemistry ; Galantamine ; chemistry ; Lycoris ; chemistry ; growth & development ; Phenylalanine ; chemistry ; Plant Cells ; chemistry ; Plant Extracts

Objective To explore the relationship between the levels of Cu, Fe, Mn, Zn in serum and Parkinson disease (PD). Methods A total of 40 patients with PD (PD group) and 40 control subjects (control group)were enrolled in this study. Serum levels of Mn were measured by graphite atomic absorption, and Cu ,Fe, Zn were measured by inductively coupled plasma(ICP)mass spectrometry. Resudts There were significantly increase in the levels of Mn and Fe in PD group than those in control group [(0.269±0.326) μ mol/L vs (0.125±0.054) μmol/L, P< 0.05, (1.512±0.949) μmol/L vs (0.676±0.111) μmol/L, P< 0.01)]. There were no significant difference in the levels of Cu and Zn between two groups (P> 0.05). Condusion Micreelements may play important roles in pathogenesis and development of PD, especially Fe and Mn.

BACKGROUNDIt has been proved that EphB4 and HIF-1α are closely related to the oncogenesis and development of lung cancer. The aim of this study is to investigate the biological significances of EphB4 and HIF-1α in lung cancer and their relationship with each other.

METHODSThe expression of EphB4 and HIF-1α was detected in 54 lung cancer tissues and 10 normal lung tissues as control by immunohistochemical method.

RESULTSEphB4 and HIF-1α proteins were detectable in 50.0% and 42.6% of all 54 lung cancer tissues respectively, which were significantly higher than those of the control (P < 0.05); the positive ratios and the levels of the expressions of EphB4 and HIF-1α proteins were closely related to gross types, differentiations and clinical stages (P < 0.05), but not to histological classification, age, sex and lymph node metastasis (P > 0.05). A highly positive correlation was observed between EphB4 and HIF-1α expression (P < 0.01 ).

CONCLUSIONSOverexpression of EphB4 and HIF-1α may play an important role in the pathogenesis, progression and malignant degree of lung cancer. Detection of EphB4 and HIF-1α expression might be helpful to predict prognosis of patients with lung cancer.

Causes analysis of death is the most common work in forensic pathology practice. When designing problem-based learning (PBL) teaching objectives, we should take cause analysis of death as the main line and give consideration to other related issues. The selected cases should be typical ones that solve the target problems, which can fully reflect the basic theoretical knowledge of forensic pathology, and have moderate difficulty, delights and clear conclusions. The PBL course is divided into three steps. The first step focuses on providing students with case information to guide them to find out the problems that need to be solved. The second step focuses on discussing the problems and making pathological diagnoses. The third step focuses on answering the question raised at the beginning of the course. Each lesson can also be divided into several sections by which the lesson plans should be prepared. In the teaching process, performing active interaction with students, controlling the direction of classroom development, balancing student opportunities should be done well in order to make the curriculum smoothly and achieve the purpose of teaching.