Objective To prepare the monoclonal antibody against human perforin(HP). Methods Recombinant eukaryotic expression plasmid pCDM8-HP was extracted and purified, and the BALB/C mice were immunized with the plasmid. The hybridomas producing anti-HP McAbs were established by using hybridoma technique, then the specificity of the McAbs was identified by using immunocytochemical technique and Western blot. Results Three hybridoma cell lines secreting McAbs against human perforin were established, and the three McAbs showed positive only with LAK cells containing human perforin protein, and showed negative with inactive human peripheral blood lymphocytes (PBLs). The subclasses of the three McAbs were determined as lgG2bK. Western blot results showed that the three McAbs recognized a specific band of LAK cell lysateds with molecular weight of 70.0Kd. Conclusion The three hybridoma cell lines secreting McAbs against human perforin were established and the secreted McAbs were specific.

Objective To express human B7.2 extracellular domain with prokaryote expression system and to evaluate its biological activity in vitro. Methods PCR was used to amplify the extracellular region of human B7. 2which contained both the IgV and IgC domains. The recombinant PGEX-4T-3/hB7. 2 (IgV+C) was obtained by cloning the PCR product into a prokaryote expression plasmid PGEX-4T-3 and was transformed into the host strain of DH5-α. The fusion protein consisted of GST and hB7.2(IgV+C) was identified by SDS-PAGE and Western blotting.T cell activation was observed by exposing purified T lymphocytes to the fusion protein and [3H]-TdR incorporation with the presence of the first signal imitated by anti-CD3 antibody. Results The fusion protein GST-hB7.2 (IgV+C) was produced and detected in inclusive body form from engineered bacterial cells. With the first signal existed,T lymphocytes proliferated when it was co-stimulated by the fusion protein. Conclusion These results indicated that the functional human B7.2(IgV+C) fusion protein can be produced in bacterial cells and the fusion protein displays the co-stimulatory activity in T lymphocytes activation.

Objective To investigate the effect of computer-assisted cognition training on the cognitive functioning of schizophrenic patients.Methods One hundred and four persons with schizophrenia were randomly divided into a research group of 51 cases and a control group of 53.The former were given computerized cognition training covering attention,memory and executive functions.The training lasted 2 months.The controls received regular occupational and recreational therapy.Before and after the treatment the 2 groups were evaluated using the Positive and Negative Syndrome scale (PANSS),the Trail Making tests A and B (TMTs),Wechsler's memory scale (WMS) and the Wisconsin card sorting test (WCST).Results The average total PANSS score of the research group declined significantly from (70.23 ±7.62) before treatment to (59.82 ± 8.41) after treatment;so did that of the control group from (68.32 ± 10.14) to (59.45 ± 9.05).The average negative symptoms scale score of the treatment group and the control group also decreased significantly for (21.46 ± 3.26) before treatment to (16.67 ±3.20),and from (20.82 ±4.21) to (17.08 ±3.23) respectively,but there was no significant difference between the groups in the magnitude.The research group's average TMT A and TMT B scores also decreased from(51.76±21.18) and (114.31 ±35.76) to (44.14 ±17.51) and (96.47 ±34.43),while no significant difference was observed before and after the treatment in the control group.The WMS recognition and regeneration scores of the research group improved significantly from (5.83 ± 3.12) and (8.63 ± 3.45) to (8.21 ± 3.96) and (10.24 ±3.52) after intervention,while no improvement was found in the control group.The same trend was observed in the total test number,the continuous error numbers,the correct numbers and the classification number scores in the WCST.Conclusion Computerized cognition training can improve the cognition of persons with schizophrenia to some extent.

AIM: The present study was designed to observe the effect of [D- Arg1, D- Trp7,9, Leu11] - substance P (spantide), a non- selective antagonist of NK receptors, on the up- regulation of nitric oxide synthase (NOS) induced by formalin test. METHODS: Formalin (5%, 0.2 mL) was subcutaneously injected into the plantar side of the right hind paw to produce persistent pain and hyperalgesia. The pain response was determined by spontaneous flinch reflex test. NOS expression was examined using NADPH- d histochemical staining. Spantide was intrathecally injected via L5 - L6 intervertebral space 5 min prior to the formalin injection. RESULTS: Injection of formalin resulted in a characteristic behavioral response consisting of vigorous scratching, biting, licking and lifting of the injected hind paw from the box' s bottom. Following these behavioral responses, the NOS expression was up- regulated in the pericentral canal region of the L5 segment of the spinal cord. Pre- treatment with spantide depressed the spontaneous flinches of the injected paw in the second phase of the formalin test. At the same time, the upregulation of NOS was substantially inhibited. CONCLUSION: It might be concluded that substance P played an important role in the up - regulation of NOS in the pericentral canal region of the spinal cord in the formalin test.

Objective To investigate the effect of nuclear factor erythroid-2-related factor 2(Nrf2) on the anti-hypoxia and anti-apoptotic ability of mesenchymal stem cells(MSCs). Methods Human embryonic kidney cells(293FT) were transfected with recombinant plasmid which overexpressed Nrf2 and helper plasmid. High-titer lentivirus which overexpressed Nrf2 were obtained. MSCs were transfected with lentivirus with Nrf2 overexpression and empty lentiviral vector to establish Nrf2-MSCs which stably overexpressed Nrf2 (Nrf2 overexpression group) and green fluorescent protein (GFP)-MSCs(control group). The expression of green fluorescent in 2 groups was observed by fluorescence microscope. The expression level of Nrf2 protein in 2 groups was measured by Western Blot. The anti-hypoxia ability of 2 groups was observed by light microscope. The anti-apoptotic ability of 2 groups was measured by flow cytometry. Results Nrf2-MSCs which stably overexpressed Nrf2 were successfully established. Western Blot analysis revealed that the expression level of Nrf2 protein in the Nrf2 overexpression group was significantly higher than that in the control group(P<0.01). After 15 h hypoxia treatment, the cell activity in the Nrf2 overexpression group was significantly higher than that in the control group. Flow cytometry showed that the apoptosis rate in the Nrf2 overexpression group was (30.9±1.4)%, significantly lower than (61.3±1.3)% in the control group(P<0.05). Conclusions Nrf2-MSCs which can stably overexpress Nrf2 possess certain anti-hypoxia and anti-apoptotic ability in hypoxia environment.

Objective: To investigate the efficacy of Linglianhua granule in the treatment of depression after coronary heart disease with syndrome of heart-kidney imbalance and its effect on serum levels of interleukin(IL)-8, IL-17, norepinephrine(NE), and 5-hydroxytryptamine(5-HT). Methods: From December 2016 to December 2018, 260 cases with depression after coronary heart disease were selected in Kangning Hospital Affiliated to Wenzhou Medical University and randomly divided into the control group and the observation group according to the digital table, with 130 cases in each group.Both two groups were given routine therapy.The control group was treated with lorazepam tablets.Based on the control group, the observation group was given Linglianhua granule (1 bag/time and bid). The two groups were treated for 8 weeks.The scores of Hamilton's depression scale(HAMD)-17, self-rating depression scale (SDS), clinical efficacy, and serum levels of IL-8, IL-17, NE, 5-HT were compared between the two groups. Results: After treatment for 8 weeks, the HAMD, SDS score of the observation group were obviously lower than those of the control group(t=7.903, 12.331, all P=0.00). The total effective rate of the observation group(93.85%) was higher than that of the control group(84.62%)(χ²=4.843, P<0.05). After treatment for 8 weeks, the serum levels of IL-8 and IL-17 in the observation group were lower, while the NE and 5-HT levels were higher, compared with those in the control group(t=9.403, 14.439, 19.431, 28.931, all P=0.00). Conclusion Based on the routine therapy, Linglianhua granule in treating depression after coronary heart disease with syndrome of heart-kidney imbalance can promote improvement of depression symptoms, increase the efficacy.Inhibiting serum levels of IL-8, IL-17 and up-regulation of NE and 5-HT may be related with the efficacy.