The aim of this study is to explore the immunogenicity of African swine fever virus p37 recombinant protein in mice.C57BL/6J mice were immunized subcutaneously in the abdomen using p37 recombinant protein as antigen.The second immunization was performed 21 d after the first immunization.Serum-specific antibody levels were detected by ELISA;serum cytokine levels were detected using a multifactor assay technique;mice splenic lymphocytes were isolated 7 d after sec-ondary immunization,and the number of splenic lymphocytes secreting IFN-γ after recombinant protein stimulation was detected by ELISpot;and the ratio of CD4+T cells to CD8+T cells was detected by flow cytometry.The results of indirect ELISA showed that p37 recombinant protein could stimulate mice to produce high levels of specific antibodies;ELISpot showed that p37 recom-binant protein could significantly stimulate splenic lymphocytes to produce IFN-γ(P＜0.001)and activate cellular immune responses;the results of flow cytometry showed that it could signifi-cantly stimulate the differentiation of T-lymphocytes to CD4+T-lymphocytes(P＜0.001).In ad-dition,serum levels of IL-2,IL-4,IFN-γ,and TNF-α immune-related cytokines were significantly higher after the second immunization.Immunization of mice with p37 recombinant protein induced strong humoral and cellular immune responses with good immunogenicity,providing reference for the subsequent epitope identification and functional study of p37 protein and the antigen screening of ASF mRNA vaccine.

The aim of this study is to explore the immunogenicity of African swine fever virus p37 recombinant protein in mice.C57BL/6J mice were immunized subcutaneously in the abdomen using p37 recombinant protein as antigen.The second immunization was performed 21 d after the first immunization.Serum-specific antibody levels were detected by ELISA;serum cytokine levels were detected using a multifactor assay technique;mice splenic lymphocytes were isolated 7 d after sec-ondary immunization,and the number of splenic lymphocytes secreting IFN-γ after recombinant protein stimulation was detected by ELISpot;and the ratio of CD4+T cells to CD8+T cells was detected by flow cytometry.The results of indirect ELISA showed that p37 recombinant protein could stimulate mice to produce high levels of specific antibodies;ELISpot showed that p37 recom-binant protein could significantly stimulate splenic lymphocytes to produce IFN-γ(P＜0.001)and activate cellular immune responses;the results of flow cytometry showed that it could signifi-cantly stimulate the differentiation of T-lymphocytes to CD4+T-lymphocytes(P＜0.001).In ad-dition,serum levels of IL-2,IL-4,IFN-γ,and TNF-α immune-related cytokines were significantly higher after the second immunization.Immunization of mice with p37 recombinant protein induced strong humoral and cellular immune responses with good immunogenicity,providing reference for the subsequent epitope identification and functional study of p37 protein and the antigen screening of ASF mRNA vaccine.

African swine fever(ASF)is a highly contagious disease caused by the African swine fe-ver virus(ASFV).To evaluate the immunogenicity of the pp62(CP530R)protein of ASFV,the recombinant CP530R protein was expressed in HEK 293F cells transfected with the plasmid pMAL-Fc-CP530R.Six-week-old female C57BL/6J mice were immunized with 10 μg of purified pp62 protein via subcutaneous injection,followed by a booster immunization with the same dosage at 21 days(enhanced).The humoral and cellular immune responses in the mice were then assessed using ELISA,flow cytometry,and ELISpot assays.Western blot analysis confirmed that the pp62 protein was successfully expressed,with a molecular weight of approximately 118.5 kDa.ELISA results indicated that a high level of specific antibodies was detected in the immunized mice,with antibody titers reaching up to 1∶1 638 400 at 7 days after the secondary immunization.The pro-portion of CD8+T lymphocytes in the immunized mice increased compared to the control group(P＜0.05).Results from the Q-PlexTM Mouse Cytokine Screen demonstrated that the secretion levels of IFN-γ,IL-2,IL-4,and IL-10 in serum were significantly upregulated in the immunized mice following secondary immunization(P＜0.001).In summary,these findings indicate that the pp62 protein can significantly stimulate both humoral and cellular immunity in mice,laying the groundwork for further studies on the function of the ASFV pp62 protein and the identification of novel vaccine antigens for ASF.

African swine fever(ASF)is a highly contagious disease caused by the African swine fe-ver virus(ASFV).To evaluate the immunogenicity of the pp62(CP530R)protein of ASFV,the recombinant CP530R protein was expressed in HEK 293F cells transfected with the plasmid pMAL-Fc-CP530R.Six-week-old female C57BL/6J mice were immunized with 10 μg of purified pp62 protein via subcutaneous injection,followed by a booster immunization with the same dosage at 21 days(enhanced).The humoral and cellular immune responses in the mice were then assessed using ELISA,flow cytometry,and ELISpot assays.Western blot analysis confirmed that the pp62 protein was successfully expressed,with a molecular weight of approximately 118.5 kDa.ELISA results indicated that a high level of specific antibodies was detected in the immunized mice,with antibody titers reaching up to 1∶1 638 400 at 7 days after the secondary immunization.The pro-portion of CD8+T lymphocytes in the immunized mice increased compared to the control group(P＜0.05).Results from the Q-PlexTM Mouse Cytokine Screen demonstrated that the secretion levels of IFN-γ,IL-2,IL-4,and IL-10 in serum were significantly upregulated in the immunized mice following secondary immunization(P＜0.001).In summary,these findings indicate that the pp62 protein can significantly stimulate both humoral and cellular immunity in mice,laying the groundwork for further studies on the function of the ASFV pp62 protein and the identification of novel vaccine antigens for ASF.

【Objective】 To demonstrate the real incidence and clinical features of acute transfusion reactions (ATRs) and analyze its association with transfusion effectiveness. 【Methods】 The blood transfusion process of patients in the Hematology Department in our hospital from March 16, 2018 to March 16, 2019 was observed and followed up. The vital signs and clinical features were recorded, the clinical data and relevant laboratory examination results were collected, and the correlation between ATRs, clinical features and transfusion effectiveness was statistically analyzed. 【Results】 A total of 2500 transfusions were observed in the Hematology Department, out of which 138 patients developed 242 ATRs. The overall incidence of ATRs was 9.68% (95% CI, 8.52%~10.94%) and the incidence of ATRs during platelets transfusion was the highest. The clinical features of most ATRs were mild and no fatal consequences occurred. No significant difference was found in the association between transfusion effectiveness with ATRs in platelets and red blood cells transfusion (P>0.05). 【Conclusion】 None of the ATRs observed in this study resulted in serious consequences or had effect on the transfusion effectiveness. The characteristics presented in these reactions may provide certain references for clinical practice.

Objective To investigate the recurrence and survival of postoperative patients with differentiated thyroid carcinoma (DTC) aged from 25 to 59 years.Methods We retrospectively analyzed the clinical data of 36 patients with DTC treated in our hospital from 1996 to 2011,and the recurrence and survival status of the patients were recorded.Kaplan-Meier analysis was carried out to analyze factors that affect the patient's survival.Results Nine patients died of recurrence or metastasis,and the interval between the initial surgery and recurrence ranged from 22 to 46 months.The survival time of the 36 patients ranged from 34 to 135 months with a 10-year survival rate of 75.0％.Kaplan-Meier analysis showed that male patients had a significantly shorter mean survival time than female patients (χ2＝3.164,P=0.041);the median survival time of patients aged 45-59 years was obviously shorter than that of patients aged 25-44 years (χ2＝4.622,P=0.032);the postoperative survival in patients with 131I therapy was significantly longer than those who did not receive the therapy (χ2＝4.527,P=0.033),and was not affected by total excision of the thyroid gland (χ2=0.988,P＝0.320).No significant difference was found in the median survival of patients in different clinical stages (χ2=2.2132,P=0.167).Conclusion In young and middle-aged patients with DTC,postoperative recurrence is the most likely in 2 to 4 years after the surgery.Male patients at 45-59 years of age who do not receive 131I treatment are at high risks of tumor recurrence.

OBJECTIVE:To investigate clinical efficacy of intravenous drip of alprostadil combined with acupuncture in the treatment of diabetic foot ulcer and its effects on inflammatory factors levels and wound microvascular density. METHODS:A total of 72 patients with diabetic foot ulcer in our hospital during May 2014 to Sept. 2015 were divided into observation group and con-trol group according to random number table,with 36 cases in each group. Control group was given Alprostadil injection 10μg add-ed into 250 mL 0.9% Sodium chloride injection intravenously,qd,on the basis of routine treatment as insulin. Observation group was additionally given acupuncture therapy (Sanyinjiao,Zusanli,Fenglong,Yanglingquan,Yinlingquan and other acupuncture points),qd,on the basis of control group. Treatment courses of 2 groups lasted for 3 weeks. Clinical efficacy,wound healing time,fasting blood glucose,urine microalbumin,inflammatory factors,wound microvascular density as well as the occurrence of ADR were compared between 2 groups. RESULTS:The total response rate of observation group(91.67%)was significantly higher than that of control group(72.22%),and wound healing time was significantly shorter than control group,with statistical signifi-cance(P<0.05). Before treatment,there was no statistical significance in fasting blood glucose,urine microalbumin and inflamma-tory factors between 2 groups (P>0.05). After treatment,above indexes of 2 groups were significantly lower than before,and urine microalbumin and inflammatory factors of the observation group was significantly lower than the control group,with statisti-cal significance (P<0.05). 14,21 d after treatment,wound microvessel density of observation group was significantly higher those of control group,with statistical significance(P<0.05). No obvious ADR was found in 2 groups. CONCLUSIONS:Intrave-nous drip of Alprostadil combined with acupuncture can boost angiogenesis through mitigating inflammatory reactions,thus shorten the time of wound healing and enhance therapeutic efficacy.

Objective To investigate the recurrence and survival of postoperative patients with differentiated thyroid carcinoma (DTC) aged from 25 to 59 years.Methods We retrospectively analyzed the clinical data of 36 patients with DTC treated in our hospital from 1996 to 2011,and the recurrence and survival status of the patients were recorded.Kaplan-Meier analysis was carried out to analyze factors that affect the patient's survival.Results Nine patients died of recurrence or metastasis,and the interval between the initial surgery and recurrence ranged from 22 to 46 months.The survival time of the 36 patients ranged from 34 to 135 months with a 10-year survival rate of 75.0％.Kaplan-Meier analysis showed that male patients had a significantly shorter mean survival time than female patients (χ2＝3.164,P=0.041);the median survival time of patients aged 45-59 years was obviously shorter than that of patients aged 25-44 years (χ2＝4.622,P=0.032);the postoperative survival in patients with 131I therapy was significantly longer than those who did not receive the therapy (χ2＝4.527,P=0.033),and was not affected by total excision of the thyroid gland (χ2=0.988,P＝0.320).No significant difference was found in the median survival of patients in different clinical stages (χ2=2.2132,P=0.167).Conclusion In young and middle-aged patients with DTC,postoperative recurrence is the most likely in 2 to 4 years after the surgery.Male patients at 45-59 years of age who do not receive 131I treatment are at high risks of tumor recurrence.

Objective To observe the effect of the rehabilitation garden on hemiplegic stroke patients. Methods 160 cases of cerebral vascular accident patients were randomized into observation group and control group. All patients were given conventional drug treatment and conventional rehabilitation training. The observation group was given the rehabilitation garden for rehabilitation training two times a day , each time 60 minutes. Results BBS, BI, FMA, HAMA and HAMD of The two groups of patients significantly improved. The improvement ofBBS and BI in observation group was better than that in control group (P < 0.05). HAMD and HAMA score inobservation group was significantly higher than that in control group (P < 0.01), the improvement in observationgroup is better than in control group (P < 0.01), the satisfaction rate in observation group is better than in controlgroup (P < 0.01), and the hospital stay period in observation group is better than in control group (P < 0.01).Conclusion The application of rehabilitation garden has a good effect on the rehabilitation effect of the hemiplegicstroke patients.

[ ABSTRACT] AIM:To investigate the role of ATP-sensitive potassium ( KATP ) channels in the inhibitory effect of hydrogen sulfide ( H2 S) on high glucose ( HG)-induced inflammation mediated by necroptosis in H 9c2 cardiac cells. METHODS:The expression levels of receptor-interacting protein 3 ( RIP3; an indicator of necroptosis ) and cyclooxyge-nase-2 (COX-2) were determined by Western blot.The levels of interleukin-1β(IL-1β) and tumor necrosis factor-α( TNF-α) were detected by ELISA .RESULTS:After H9c2 cardiac cells were treated with 35 mmol/L glucose ( HG) for 24 h, the expression of RIP3 was significantly increased .Pre-treatment of the cells with 100 μmol/L diazoxide ( DZ; a KATP channel opener) or 400 μmol/L NaHS (a donor of H2S) for 30 min considerably blocked the up-regulation of RIP3 induced by HG.Moreover, pre-treatment of the cells with 100 μmol/L 5-hydroxydecanoic acid (5-HD; a KATP channel
blocker) attenuated the inhibitory effect of NaHS on HG-induced up-regulation of RIP3.On the other hand, co-treatment of the cells with 100μmol/L necrostatin-1 ( a specific inhibitor of necroptosis ) or pre-treatment of the cells with 100 μmol/L DZ or 400 μmol/L NaHS attenuated HG-induced inflammatory responses , evidenced by decreases in the expression of COX-2 and secretion levels of IL-1βand TNF-α.However , pre-treatment of the cells with 100 μmol/L 5-HD significantly attenuated the above anti-inflammatory effects of NaHS.CONCLUSION:KATP channels play an important role in the inhib-itory effect of H2 S on HG-induced inflammation mediated by necroptosis in H 9c2 cardiac cells.