AIM: To study the physiological function changes of the tears film after rebuilding ocular surface with corneal stem cells, and to discuss the validity and the estimate system of rebuilding ocular surface with the corneal stem cells. METHODS: The male New Zealand rabbits were used to establish the alkali burning model in the right eye. The corneal stem cells of the left eye were cultured on the amniotic membrane in vivo, and then transplanted to the right eye. Furthermore, the physiological function changes of the tear film were examined. RESULTS: Compared to the before alkali burning, the ocular surface cell morphology was similar after rebuilding ocular surface with the corneal stem cells, which were cultured on the amniotic membrane in vivo; The tear film breakup time test showed the a remarkable difference between after and before the alkali burning, but the cell modality after rebuilding had no remarkable difference compared to that before the alkali burning. CONCLUSIONS: It's an effective method to rebuild the ocular surface with the corneal stem cells in vivo, the cell framework and the physiological function of the tears film recover well after rebuilding. It may be a validity estimate system of rebuilding ocular surface to analyze framework and configuration of the ocular surface and test the tear film breakup time.

Objective:To observe the clinical efficacy of electroacupuncture in the treatment of type 2 diabetes mellitus combined with mild and moderate carpal tunnel syndrome.Methods:Eighty patients with type 2 diabetes mellitus combined with mild to moderate carpal tunnel syndrome were randomly divided into two groups with 40 patients in each group ( n=40). In the course of the treatment, 2 patients in each group dropped out, which means 38 patients in each group were included. All patients were instructed to actively control blood glucose The control group was given Mecobalamin tablets orally, while the treatment group was given electroacupuncture of Daling and Neiguan on the affected side based onthe control group for 6 weeks. The degree of pain was assessed by VAS, and the severity of clinical symptoms was assessed by the symptom scale of Boston carpal tunnel questionnaire (BCTQ). The sensory nerve conduction velocity (SCV), the amplitude of sensory nerve action potential (SNAP) and the distal motor latency (DML) were detected by EMG evoked potential instrument. The clinical efficacy was evaluated and adverse events were recorded. Results:The total effective rate was 81.6% (31/38) in the treatment group and 60.5% (23/38) in the control group, and the difference was statistically significant ( χ2=5.094, P=0.024). After the treatment, the scores of VAS and BCTQ in the treatment group were lower than those of the control group ( t=2.639, 2.790, P=0.010, 0.007); DML (3.62 ± 0.19 ms vs. 4.00 ± 0.17 ms, t=68.891) in the treatment group was lower than that of the control group ( P<0.01), while SCV (47.36 ± 0.18 m/s vs. 42.34 ± 0.14 m/s, t=97.163) and SNAP (14.74 ± 0.18 μV vs. 12.10 ± 0.16 μV, t=51.434) in the treatment group were higher than those of the control group ( P<0.01). During the treatment, no serious adverse events occurred in two groups. Conclusions:Electroacupuncture is safe and effective in the treatment of mild and moderate carpal tunnel syndrome with type 2 diabetes. It can reduce the degree of pain and improve the symptoms of wrist joint.

Objective NKT cells are very important as a kind of non-specific immune cells. Much attention in antitumor significance has been received in the study of its effect on malignant diseases. The aim of this study was to detect the expression of NKT cells and its CD+8 NKT subsets in peripheral blood of esophageal patients and normal person, and to analyze the changes in the expression of NKG2D and NKG2A receptorsand its clinical pathological factors. Methods By flow cytometric analysis, 53 patients with esophageal carcinoma and 39 normal controls were analysed for peripheral blood of NKT cells and CD+8 NKT subsets, and the expression of NKT cells NKG2A and expression of NKG2D receptor. The clinical pathological factors were collected for the comparative analysis. Results Compared with the normal control group, the expression of NKT cells in peripheral blood of esophageal patients increased [(4.32±0.73) %, (5.97±1.29) %] (t =3.562, P <0.01), and the expression level of its surface receptor NKG2D reduced [(17.56±5.92) %, (15.12±1.56) %] (t =3.892, P <0.05), but the express levels of NKG2A [(4.02±1.41) %, (5.99±4.59) %] in creased (r = 4.015, P <0.05), those expression change with the development of the esophageal cancer. Conclusion The increased expression of NKT cells and CD+8 NKT subsets in the peripheral blood of patients with esophageal carcinoma refletcs that the immune feedback of patients' antineoplastic effect is strengthened. The decresed expression of the active receptor NKG2D and the increased expression of the inhibitory receptor NKG2A on NKT cells might be one of mechanisms leading to the reduction of NKT cell activity and immune escape of patients with esophageal carcinoma. The changes of surface receptors of NKT cells may be associated with the development of the esophageal cancer.

Objective To investigate the application of strain rate imaging in quantitative assessment of left ventricular regional diastolic function after intracoronary stent implantation.Methods Fifty-six healthy person and 60 patients with coronary artery disease were performed quantitative assessment of left ventricular regional diastolic function by apical four chamber view,apical two chamber view and left ventricular long axis view before and after intracoronary stent implantation using strain rate imaging.Results Compared with those of normal myocardium,the peaks of strain rate curve at diastole of ischemic myocardium were reduced,and were increased after operation (P ＜ 0.05).Conclusions Intracoronary stent implantation can significantly improve the blood supply to the ischemia myocardium.Strain rate imaging can quantitatively analyze the changes of left ventricular regional diastolic function.

AIM: To establish HPLC-digitized fingerprint spectrum (HPLC-DFPS) of Rhizoma Pinelliae so that it will be applied in quality control of Rhizoma Pinelliae. METHODS: A gradient separated method was applied. Column: Agilent Zorbax ODS C_ 18 ; Mobile phase: acetonitrile-water; Detection-wavelength: 210 nm; Flow rate: 0.5 mL/min; Column temperature: 25℃. RESULTS: Using above optimum chromatograph condition, the HPLC-DFPS and technic parameters of Rhizoma Pinelliae were established. CONCLUSION: Comparison of a group of characteristic peaks suggests that the HPLC-DFPS technology is feasible in quality control of Rhizoma Pinelliae.

Object To investigate the berberine in Coptis chinensis Franch. processed with various quantity of Evodia rutaecarpa (Juss.) Benth. and the components absorbed from E. rutaecarpa. Methods Taking berberine, evodiamine and rutaecarpine as targets, HPLC method was used to determine the components of C. chinensis, E. rutaecarpa, C. chinensis processed with 10%, 20%, 30%, 40%, 50% E. rutaecarpa. Results The content of berberine in C. chinensis processed with E. rutaecarpa decreased, and that of C. chinensis processed with 20% and 30% E. rutaecarpa was higher than the rest. C. chinensis processed with E. rutaecarpa absorbed the components of E. rutaecarpa really. Conclusion The suitable quantity of E. rutaecarpa is 20% when processing for C. chinensis.

Objective To investigate the protein and gene expression of bcl-2, bcl-6 in diffuse large B-cell lymphoma (DLBCL). Methods 73 cases of DLBCL were selected for study using the Envision immunohistochemistry method with a panel of antibodies CD3, CD10, CD20, bcl-6, bcl-2, MUM-1. The bcl-2gene expression in 57 of 73 cases with chromosome translocation t (14; 18), breakage and amplification of 3q27 chromosome in 54 of 73 cases were detected by fluorescence in situ hybridization (FISH) method. Results The percentages of tumor cells expressing CD10, bcl-6, MUM-1, bcl-2 were separately 15.1%, 38.4 %, 71.2 %, 79.2 %. t (14;18) chromosomes were detected in 16 of 57 cases (28.1%). The expression of bcl-2 protein have significantly correlated with immunophenotype subtype (P=0.035), and t (14;18) was significantly correlated with the prognosis (P=0.045). There were no association between the expression of bcl-2 protein and t (14;18)(P=0.710). 11 of 54 cases were presented with 3q27 chromosomal breakage (20.4 %), and 14 cases were chromosomal amplification (25.9 %). The prognosis of cases with positive bcl-6 protein was better than that with negative protein obviously. There was no relationship between bcl-6 and 3q27 chromosomal breakage or amplification (P=1.000). Conclusion The expression of bcl-2, bcl-6 protein and gene were different events and had the different significance on DLBCL. The expression of bcl-2 protein was a prognostic marker correlated with immunophenotype subtype, and GCB type with the positive expression of bcl-2 protein had the poor prognosis. Conversely, t(14;18) was an independent event for the prognosis, and the positive expression have the poor prognosis. Patients who require the target therapy should be detected for the t(14;18). The expression of bcl-6 protein was beneficial to the judgment of DLBCL prognosis, it could be an independent factor of the prognosis. 3q27 chromosomal breakage may be a hint to the poor prognosis.

Objective To analyze CD+4 CDHi25 CDLo127 regulatory T cells (Treg) in peripheral blood of patients with thyroid nodules and differentiated thyroid cancer and their change regularity, and to investigate the immunosuppression mechanism. Methods The peripheral blood was collected from 175 patients with thyroid nodules, including 43 patients with differentiated thyroid cancer and 132 patients with nodular goiter.By using monoclonal antibodies, the blood samples were evaluated with the flow cytomertry for lymphocyte subsets and Treg cells. Results The results showed the prevalence of the CD+4 CDHi25 CDLo127 Treg in differentiated thyroid cancer group [(6.48±1.49) %] and nodular goiter group [(6.23+1.67) %] was higher than those in the healthy group [(5.62±1.48) %], and the difference was significant(P < 0.005), but there was no significant difference between the nodular goiter group and differentiated thyroid cancer group (P >0.005).Conclusion It is concluded that the relative increase of CD+4 CDHi25 CDLo127 Treg in peripheral blood of patients with nodular goiter and differentiated thyroid cancer may be related to immunosuppression and tumor progression.

Objective To observe the influence of Ligustrazine on the hemodynamic effects and prognosis in elderly patients with early cerebral ischemia following intracranial aneurysm operation ,and explore the therapeutic value .Methods 43 elderly patients with ECI following intracranial aneurysm operation were randomly divided into the Ligustrazine group (20 cases) ,and the conven-tional treatment group(23 cases)according to the table of random numbers .The patients in the Ligustrazine group were given intra-venously Ligustrazine injection and combined with conventional therapy ,whereas ,other patients were given conventional treat-ments .Results The clinical effective rate in Ligustrazine group was significantly higher than conventional treatment group (P<0 .05) .Patients recovered well in conventional treatment group were 7 cases(30 .43% ) ,mild disability 8 cases(34 .78% ) ,severe dis-ability 4 cases(17 .39% ) ,plant survival 1 case(4 .34% ) ,3 cases died(13 .06% ) ,data peer-based in the Ligustrazine group were 12 cases(60 .00% ) ,5 cases(25 .00% ) ,2 cases(10 .00% ) ,0 case and 1 case(5 .00% ) ,which were significantly lower than that of con-ventional treatment group .The prognostic scores in Ligustrazine group were apparently higher than conventional treatment group by using Glasgow outcome scale and Chinese stroke scale(P<0 .05) .Meanwhile ,the indicators of hemodynamic effects also reflec-ted that the Ligustrazine improved the blood circulation of patients with ECI ,which was better than the conventional treatment group(P<0 .05) .Conclusion The Ligustrazine Phosphate for injection showed better curative effect and prognosis in elderly pa-tients with ECI following intracranial aneurysm operation .

Background and purpose:Cytotoxic T lymphocyte (CTL) plays a vital role in the process of anti-tumor immunology. The aim of this study was to investigate whether changes in concentration of IL-2 (50, 200 and 1 000 U/mL) would affect the sub-population and cytotoxic function of cells cultivated by peptide-specific CTL induction systemin vitro and also observe whether using the concentration of IL-2 at a range of 50-1 000 U/mL isbeneifcial to regulatory cells (Tregs) enrichment.Methods:Peripheral blood from 10 healthy donors and 10 cancer patients that were HLA-A2 positive, were collected in the study. HLA-A2 restricted CTL epitope P321 (ILIGETIKI) derived from COX-2 pulsed with different concentrations of IL-2 were used to induce peptides-speciifc CTLin vitro. Flow cytometry was performed to analyze the proliferative capability, the proportion of different T-cell subsets, and secretion of perforin, granzyme B and IFN-γ. IFN-γ secretion was assessed by ELISpot assay.Results:High concentration of IL-2 increased the proliferative activity. The percentage of CD4+ T cells of cancer patient group was signiifcantly higher than that of healthy donor group, while the percentage of CD8+ T cells of cancer patient group was signiifcantly lower than that of healthy donor group. And there was no signiifcant difference in the percentages of CD4+ T cells, CD8+ T cells and Tregs among groups with different IL-2 concentrations. No difference was seen in cytokine (perforin, granzyme B, IFN-γ) secretion capacity of CD8+ T cells. ELISpot study revealed that high-dose IL-2 resulted in the increasing of IFN-γ secretion.Conclusion:The sub-population and the function of cells cultured by peptide-speciifc CTL induction systemin vitro are not affected by different concentrations of IL-2. Furthermore, high concentrations of IL-2 (50-1 000 U/mL) do not provide the enrichment for Tregs. Higher concentration of IL-2 is likely to cause high secretion of IFN-γ in ELISpot assay. In order to exclude the distraction of NK cells or NKT cells, the concentration of 50 U/mL is better choice.