Objective To establish chemiluminescent immunoassay(CLIA) for quantitative detection of Rabies virus antibody in human serum.Method The purified antigen was coated on the microplate,pairing with alkaline phosphatase(ALP)-labeled antigen and luminescence substrate CSPD,then the double antigen sandwich CLIA was established.Results The sensitivity of the assay was 0.2IU/ml.The linear range was from 0 IU/ml to 200 IU/ml.The precision was less than 10%.The analytical recovery rate was from 90% to 110%.Conclusion The CLIA is a simple,sensitive,special and repeatable method for detection of Rabies virus antibody.It was suitable for clinical application.

Background Perineural invasion is an important biological character for adenoid cystic carcinoma (ACC) of lacrimal gland,which is different from those of other lacrimal gland tumors.As the important part of neurotrophic factors,glial-derived neurotrophic factor (GDNF) plays an important role in perineural invasion for ACC of salivary gland.GDNF regulation in the ACC cell biology function needs to be further explored.Objective This study was to investigate the effect of GDNF on proliferation and migration of ACC cells,and to explore the mechanism of neural invasion in ACC of lacrimal gland.Methods ACC-2 cell line was cultured and passaged in RPMI 1640 medium with 10％ fetal bovine serum,100 U/ml penicillin and 0.1 g/L streptomycin.Single-cell suspension was prepared with the density of 2×104/ml using logarithmic phase of cells and then incubated to 96-well plate.GDNF with the final concentration of 20,60,80,100 and 120 μg/L was added into the medium respectively in the experimental groups,and the cells were cultured in the medium without GDNF as the control group.The expression of GDNF in ACC-2 cells was detected by immunohistochemistry.MTT assay was employed to assay the absorbance value at the wavelength of 570 nm (A570) for the evaluation of proliferation of ACC-2 cells after cultured by different concentrations of GDNF for different time points.Meanwhile,transwell chamber was used to examine the cell migrated number.Results Immunochemistry assay exhibited that ACC-2 cells showed the positive response for GDNF with the brown staining in the cytoplasm.In 48 hours after culture,the A570 value was elevated with the increase of GDNF concentration,showing a significant difference among various groups (F =3.336,P =0.026),and the A570 value in various concentrations of GDNF groups was higher than that of 0 μg/L GDNF group (all P＜0.05).After action of 80 μg/L GDNF,the A570 value of the cells was gradually increased with the prolong of culture time (Ftime =39.979,P=0.000).In 30 minutes after GDNF cultured,the number of migrated cells increased with the increase of GDNF concentration (F=144.886,P=0.000).ACC-2 cells were cultured by 100 μg/L GDNF for 24,30 and 40 hours,the number of migrated cells were more as the time lapse,and more migrated cells were seen in GDNF group at various time points (Ftime =46.747,P =0.000 ; Fgroup =63.786,P =0.000).Conclusions GDNF can stimulate the proliferation and migration of ACC-2 cells in a dose-and time-dependent manner.

AIM　To study the effects of dexamethasone on expressing monocyte chemoattractant protein-1(MCP-1 ) mRNA in the rats with pulmonary fibrosis, elaborate the molecular mechanism of dexamethasone (Dxs) in pulmonary fibrosis therapy. METHODS　The model of pulmonary fibrosis was established by instilling bleomycin intratracheally. After treating with Dxsip, the levels of MCP-1 mRNA were determined by RT-PCR. The histological changes were observed and the numbers of inflammatory cells were counted in optical microscopy field. RESULTS　The accumulation of inflammatory cells decreased markedly, and the symptom of pulmonary fibrosis was alleviated. Furthermore, Dxs evidently inhibited the expression of MCP-1 mRNA in lung tissues with pulmonary fibrosis. CONCLUSION　The molecular mechanism of Dxs in pulmonary fibrosis therapy was associated with inhibiting the expression of MCP-1 mRNA.

BACKGROUND:Mesenchymalstem cels have pluripotent differentiation, and can promote cel engraftment and immune regulation. Therefore,we attempt to use human umbilical cord mesenchymal stem cels as anew source for treatment of lung cancer by exploringcelisolation, identification and transplantation combined with chemotherapyforlung cancer in mice.
OBJECTIVE:To investigate the isolation and identification of human umbilical cord mesenchymal stem cels and its transplantation combined with chemotherapy for lung cancer inmice.
METHODS:Human umbilical cord mesenchymal stem cels were isolated from fresh umbilical cord of newborns and identified using tissue culture and enzyme digestion. Twenty Balb/C nude mouse models of lung cancer were randomly divided into two groups:mice in chemotherapy group were given chemotherapy, and those incombinedgroup given combination of chemotherapy with human umbilical cord mesenchymal stem cel transplantation.
RESULTS AND CONCLUSION:Compared with the chemotherapy group, the gastrointestinal tract was rosy and shiny, intestinal mucosa was smooth and complete, and tumor mass and blood indexes significantly decreased in thecombinedgroup (P< 0.05). To conclude, mature human umbilical cord mesenchymal stem cels can be obtained by tissueculture and enzyme digestion, andthecel transplantation combinedwith chemotherapy can significantly reduce gastrointestinal tract damage and themake peripheral hemogram in a stable level.

BACKGROUND:So far the positive or negative effects of mesenchymal stem cel s on tumor growth and metastasis are under discussion. OBJECTIVE:To explore the mechanism of bone marrow mesenchymal cel s in promoting lung cancer metastasis. METHODS:Primary rat bone marrow mesenchymal stem cel s were obtained by direct adherence method of the whole bone marrow, and differential adherence combined with digestion control method was performed to purify cel s. Lung cancer cel lines were cultured, and the effects of bone marrow mesenchymal stem cel s on the migration, invasion and metastasis of lung cancer cel s were observed by scratch test, cel invasion and migration assays. Orthotopic lung cancer models were established in rats and bone marrow mesenchymal stem cel s were seeded onto the left lung of rats. Then, pathological changes of lung tissues were observed at 14 days after transplannation. RESULTS AND CONCLUSION:After the scratch test, the migration rate of lung cancer cel s became higher, and the scratches healed with time. And after cel transplantation, the number of migrated lung cancer cel s increased, as wel as the ability of lung cancer cel s penetrating the Matrigel was strengthened. Besides, fibrous connective tissues could be found around the lung cancer tissues, and necrosis with distinct boundary and large tumor nuclei;the metastatic tissues showed obvious infiltration and necrosis with large tumor nuclei. These results suggest that bone marrow mesenchymal stem cel s can promote the invasion, migration and metastasis of lung cancer cel lines.

Objective To detect the expression of matrix metalloproteinase-9 (MMP-9) gene in peripheral blood of patients with oral paraquat (PQ) poisoning and evaluate its predictive value for their prognosis.Methods Thirty-seven cases of oral PQ poisoning admitted to Linyi People's Hospital from January 2013 to June 2014 were enrolled,and they were divided into survival group (26 cases) and death group (11 cases) according to the survival sitnation in 28 days after poisoning;a healthy control group included 10 healthy people selected in the same period.The peripheral blood 3 mL was collected from each PQ patient on the 1st and 3rd day after admission,and in the healthy control group,3 mL peripheral venous blood was obtained under fast on the day for physical examination.The MMP-9 gene expression of peripheral blood mononuclear cells (PBMCs) was detected by reverse transcription-polymerase chain reaction (RT-PCR) methods;the serum MMP-9 concentration was determined by using enzyme-linked immunosorbent assay (ELISA);the serum PQ level was detected by high performance liquid chromatography (HPLC),and the amount of poison orally taken was recorded.The correlations between PQ amount orally taken,serum PQ level and MMP-9 expression were analyzed by Spearman correlation analysis.A receiver operating characteristic (ROC) curve was drawn to evaluate the predictive value of peripheral blood MMP-9 level for the 28-day prognosis of PQ poisoning patients.Results After admission the 1 day serum PQ level was (2.60 ± 1.29) mg/L,and the amount of poison taken was 50.0 (7.5,60.0) mL in the 37 patients with oral PQ poisoning.The MMP-9 gene expression level in PBMCs and serum MMP-9 protein level of both PQ poisoning groups were significantly higher than those of healthy control group,and the levels were gradually increased with the extension of poisoning time;the degrees of elevation in death group were more significant [the PBMCs' MMP-9 gene expression (A value):2.84± 1.16 vs.0.95 ± 0.23 on the 1st poisoning day,4.22± 1.75 vs.1.29 ±0.30 on the 3rd poisoning day;serum MMP-9 concentration (μg/L):2791.48± 1 230.88 vs.807.81±279.86 on the 1st poisoning day,4384.21 ± 1 781.97 vs.1 131.14±291.76 on the 3rd poisoning day,all P ＜ 0.05].Correlation analysis showed:there were significant positive correlations of oral PQ amount,serum PQ concentration to the MMP-9 gene expression in PBMCs and serum MMP-9 protein concentration in patients with oral PQ poisoning (all P =0.000).ROC curve analysis showed:the MMP-9 gene expression in PBMCs on the 1st day and the serum MMP-9 content on the 3rd day after admission had predictive value for 28-day prognosis in patients with oral PQ poisoning,and the ROC areas under the curve (AUC) was 0.820 and 0.776 respectively.When the cutoff value of MMP-9 gene expression level on the 1st day after admission was 0.90,the predictive sensitivity and specificity were 80.00％ and 63.64％ respectively;when the cutoff value of serum MMP-9 protein content on the 3rd day after admission was 904.36 μg/L,the predictive sensitivity and specificity were 80.00％ and 72.73％ respectively.Conclusion Oral PQ poisoning can lead to the MMP-9 gene expression in PBMCs and elevation of serum MMP-9 protein level in the body,and the MMP-9 gene expression has predictive value for the prognosis of patients with oral PQ poisoning.

·AIM: To evaluate the function of primary human corneal endothelial cells(HCEC) serving as immunological cells. ·METHODS: Expression of HLA-DP, -DQ, -DR, CD40, CD80, and CD86 was determined by immunohistochemical methods. Meanwhile, purified peripheral blood mononuclear cells(PBMC) were cocultured with primary HCEC which were pre-treated with and without γ-IFN respectively; activation of lymphocytes was determined by FACS analysis.·RESULTS: In coculture system, T lymphocyte was activated by primary HCEC, HLA-DP, -DQ, -DR and CD40 expression were increased by γ-IFN induction. Costimulatory molecular CD80 was shown on the endothelial cells. ·CONCLUSION: Primary HCEC were assumed to be involved in the corneal transplantation rejection process as potential antigen presenting cells(APC).

Objective To investigate change of chemokine mRNA and protein in the patients with diabetes mellitus(DM)complicated infection.Methods Fourty-eight patients with DM complicated infection were studied.Expression of mRNA and protein of monocyte chemoattractant protein-1(MCP-1)and interleukin 8 (IL-8)were measured by FQ-PCR,RT-PCR and ELISA.The relationship between hs-CRP,PMNL,IL-8 and MCP-1 were analyzed.Results Expression of mRNA and protein of MCP-1 and IL-8 were increased in the DM patients complicated infection.MCP-1 mRNA(mesured by FQ-PCR)was positively correlated with the protein(IL-8 r = 0.33;MCP-1 r = 0.88;P

AIM To study the effects of dexamethasone on expressing monocyte chemoattractant protein 1(MCP 1 ) mRNA in the rats with pulmonary fibrosis, elaborate the molecular mechanism of dexamethasone (Dxs) in pulmonary fibrosis therapy. METHODS The model of pulmonary fibrosis was established by instilling bleomycin intratracheally. After treating with Dxs ip , the levels of MCP 1 mRNA were determined by RT PCR. The histological changes were observed and the numbers of inflammatory cells were counted in optical microscopy field. RESULTS The accumulation of inflammatory cells decreased markedly, and the symptom of pulmonary fibrosis was alleviated. Furthermore, Dxs evidently inhibited the expression of MCP 1 mRNA in lung tissues with pulmonary fibrosis. CONCLUSION The molecular mechanism of Dxs in pulmonary fibrosis therapy was associated with inhibiting the expression of MCP 1 mRNA.

Objective To quantify the changes of tongue color with the development of the disease according to the characteristics of human vision.Methods Psychological scales of tongue color perception were obtained from thc psychophysical experiments.Polynomial regression and support vector regression (SVR) were used to establish the mathematical model between the physical values and the psychological scales of tongue color.Results The psychological scales exported from the model could correspond to the visual perception of tongue color change, and SVR has higher accuracy.Conclusions The psychological scale, can not only quantitatively evaluate the tongue color in the development of the disease, but also quantify the degree of disease, to assist the doctors for disease diagnosis and treatment.