Aim To investigate the effect and mechanism of cobra venom metalloproteinase atrase A on inhibiting platelet aggregation.Methods Platelet aggregation induced by collagen,ADP,PAF,AA,ristocetin and thrombin,respectively,was measured turbimetrically after platelet incubated with atrase A.Western blot was used to detect the effect of atrase A on cleavage of platelet membrane glycoprotein and von Willebrand Factor.Results Atrase A significantly inhibited platelet aggregation induced by ristocetin and thrombin in a dose-and time-dependent manner.Meanwhile,atrase A just showed slight inhibitory effect on platelet aggregation induced by PAF,AA,collagen,and ADP after incubated with PRP for 5 min.After incubation time was prolonged to 30 min,significant inhibition was shown on platelet aggregation.Western blot revealed that atrase A cleaved platelet membrane glycoprotein GPIb.Conclusions Cobra venom metalloproteinase atrase A significantly inhibits platelet aggregation induced by ristocetin and thrombin due to the cleavage of platelet membrane glycoprotein Ib.And atrase A also has inhibitory effect on platelet aggregation induced by ADP,PAF,AA,and collagen.

Aim To investigate the inhibitory effect of atrase B on human platelet aggregation induced by activated complement.Methods By employing CVF to activate complement,the effect of atrase B on gel filtered platelet aggregation induced by activated complement was measured by turbidimetry and the expression of P-selectin and GPⅡb/Ⅲa on platelet membrane were detected by flow cytometry.Results Atrase B inhibited platelet aggregation and the expression of P-selectin and GPⅡb/Ⅲa on platelet membrane induced by activated complement.Conclusion Anticomplementary protein atrase B from Naja atra venom can significantly inhibit platelet activation and aggregation induced by activated complement.

Objective: To study the clinicopathologic characteristics and diagnostic criteria of primary mediastinal B-cell lymphoma (PMBL), and to distinguish PMBL from classic Hodgkin lymphoma(CHL) and systemic diffuse large B-cell lymphoma(DLBCL). Methods: The clinical features, histologic findings, results of immunohistochemical study and prgnosis in 27 PMBL cases were analyzed, with review of literature. Results: The age of patients ranged from 19 to 82 years (median age 34 years). All cases were located in the mediastinum and frequently accompanied by superior vein cava syndrome. Histologically, the tumor cells were pleomorphic and diffusely distributed. Clear cytoplasm and spindle tumor cells were seen in some cases. Varying amount of sclerosing stroma with collagen deposition was seen.Immunohistochemical study showed that the tumor cells were positive for CD20(100%, 27/27), CD30 (64.0%, 16/25), CD23 (77.3%, 17/22) and p63 (16/19). Clonal B cell gene rearrangement was seen. Conclusions PMBL is a subtype of diffuse large B-cell lymphoma with various histomorphology. Immunohistochemistry can help to confirm the diagnosis, and the prognosis is better than diffuse large B cell lymphoma, not otherwise specified.

To study the umbilical cord mesenchymal stem cells impact of inflammatory factors in rheumatoid arthritis patients.Methods:94 cases of patients with RA hospitalized in our department from April 2011 to December 2012 were treated with umbilical cord mesenchymal stem cells (MSCs) UC,during which the cell therapy scholastic ethics was committed approvally and patients′informed consents were separately signed .94 patients were directly intravenous infusion with 40 ml UC-MSCs ( 4×107 cells/ml),including 57 cases with two UC-MSCs therapy.We used multifunctional flow lattice Luminex 200 analysis to detect contents of 13 kinds of factors in serum such as TNF alpha,IFN-gamma,IL-1β,IL-4,IL-6,IL-8,IL-10,ie,and detected CPR,ESR,assessment DAS28,HAQ,and ARA.Follow-up treatments were performed after 1 week,3 months,6 months ( two cells treatment of 60 cases).Results:①DAS28,HAQ grading standards,were decreased (P<0.01) in 3 months and 6 months before the patients treatment ,2 times than one continue treatment decreased ( P<0.01 );the ESR and CRP in 1 week dropped significantly after treatment ( P<0.01),3 months,6 months before the treatment also decreased (P<0.05).②IL-6,TNF alpha were in falling levels after 1 week,3 months and 6 months treatment ,( P<0.05 ).Conclusion: We proved the inflammatory factors directly related with clinical indicators and symptoms of RA patients .94 cases of patients with other inflammatory factor (IL-17,IL-4,IL-10,etc.) also had some change,we still needed further observation.According to drug rheumatism guide at the same time , collaborative using with UC-MSCs could make RA patients improve local and systemic inflammatory response ,prevent disease progression.

Objective:To investigate the pahtogengesis and pathology of delayed xenograft rejection (DXR) after pig to rhesus monkey heart xenotransplantation.Methods:Heterotopic xenogeneic heart transplantation in the abdominal cavity was performed using piglet as donors.4 monkeys were used as recipients.Complete complement depletion was achieve in the recipients treated with repetitive doses of a high activity cobra venom factor (Y CVF).The recipients were immunosuppressed with a combination of cyclosporine A,cyclophosphamide and steroids.Sera were analyzed for C3,C4 levels and complement activity and anti pig endotheliocyte xenoantibody.The graft were examined histopathologically and immunohistochemically for C3,C4,C5b 9,IgM,IgG,necrosis factor alpha (TNF alpha),intercellular adhesion molecule 1 (ICAM 1),CD57 (NK cells),CD68 (macrophages),CD4 and CD8.Results:The xenografts survived 8,10,13,13 days respectively and all grafts occoured DXR.Venular thrombosis was outstanding feature within DXR xenografts,complicated with interstitial edma,local hemorrhage and myocardial necrosis,with mild to moderate cellular infiltration.The serum C3 levels and complement activity almost decreased to 0 from the day of transplantation due to Y CVF,the C4 level began to decrease 2 4 days before the cardic xenografts losing their function.The anti pig endotheliocyte xenoantibody also decreased after transplantation,and slightly increased during DXR,all rejected xenografts showed C3,C4,C5b 9,IgG and IgM deposits in different degree.Large numbers of macrophages (50% of total leukocytes) were found infiltrating the entire xenograft,a few natural killer cells (8%～10%),some of CD4+T cells (15%) and CD8+T cells (25%) were detcted also,up regulation of ICAM 1 on the graft endothelial cells and TNF alpha in the interstitial were demonstrated in the rejected heart.Conclusion:Both Humor and cell mediated immunologic reaction may play an important role in pahtogengesis of DXR. [

Objective To study the correlation between MGMT expression , clinicopathologic features and post-chemotherapy prognosis in patients with primary central nervous system lymphoma diffuse large B-cell lymphoma (PCNS-DLBCL).Methods MGMT expression was detected in 76 cases of PCNS-DLBCL by EnVision method with immunohistochemical staining .Follow-up data including treatment response and overall survival time , were analyzed.Results The rate of MGMT expression in PCNS-DLBCL was 67.1%(51/76).The MGMT expression rate in male patients was higher than that in female (P<0.05). Univariate analysis showed that these clinical pathological characteristics affected the overall survival of PCNS-DLBCL patients , including age and Hans algorithm , although no statistical significance was detected ( P value was 0.065 and 0.069 respectively ) .The overall survival of the patients with positive MGMT and aged over 60 years was shorter after chemotherapy than those without chemotherapy ( P=0.022 ) .In the patients aged over 60 years, the prognosis of MGMT-positive patients was significantly better than MGMT-negative patients ( P=0.044 ) .Conclusions The expression of MGMT is more commonly found in male patients.In the patients aged over 60 years with the same therapy , the prognosis is better in the MGMT-negative ones.Detection of MGMT protein expression can provide some guidance in choice of treatment modalities in PCNS-DLBCL patients.

Objective:To investigate the diagnostic value of preferentially expressed antigen in melanoma (PRAME) immunohistochemical staining in differential diagnosis of primary endometrial and endocervical adenocarcinomas.Methods:Eighty‐seven cases of endometrial adenocarcinoma and sixty-three cases of cervical adenocarcinoma were collected from May 2018 to November 2023 in the Department of Pathology, Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School and all the cases were subject to PRAME immunohistochemical staining. The difference of PRAME expression between endometrial and endocervical adenocarcinomas was analyzed.Results:In 87 cases of endometrial adenocarcinoma, patients′ age ranged from 35 to 71 years (average 59 years, median 59 years); in 63 cases of cervical adenocarcinoma patients′ age ranged from 28 to 80 years (average 49 years, median 47 years). Seventy‐eight cases (78/87, 89.7%) of endometrial adenocarcinoma; 2 cases (2/63, 3.2%) of cervical adenocarcinoma showed positive PRAME staining, and both cases of cervical adenocarcinoma were clear cell carcinoma. The sensitivity and specificity of PRAME in distinguishing between endometrial and cervical adenocarcinoma in the cohort were 89.7% and 96.8%, while those in differentiating non-clear cell carcinoma of the uterus from that of the cervix reached up to 91% and 100%, respectively.Conclusions:Immunohistochemical staining for PRAME demonstrates statistically significant differences between endometrial and cervical carcinomas, making it a useful auxiliary diagnostic marker for differentiating cervical and endometrial adenocarcinoma, especially non-clear cell carcinoma.

The study adopted muscle injection of pVAX1-SP-GHRH(1-44)expression plasmid and electrostimulation to determine its effects on mouse growth and sow production performance.One hundred and fifty four-week-old C57 BL/6 female mice were randomly divided into 6 groups of 5 replicates each.Muscle single-injection followed by electrostimulation was performed.The con-trol group received an empty plasmid injection(80 μg/kg),while the treatment groups received pVAX1-SP-GHRH(1-44)plasmid injections(20,40,80,120,160 μg/kg).Twenty healthy preg-nant sows were randomly divided into 2 groups,each with 10 sows.Electrostimulation treatment was applied to the semimembranosus muscle of the pregnant sows after a single injection.The con-trol group received physiological saline injection,while the plasmid group received a 2 mg pVAX1-SP-GHRH(1-44)expression plasmid injection.Mouse weight,feed intake,and serum GHRH and IGF-1 levels were measured at days 0,7,14,21,and 28 after injection.Pregnant sows were bled via the tail vein at days 0,14,28,and 42 after injection,and their serum was separated to measure serum GHRH and IGF-1 levels.The birth weight,placental weight,number of piglets born,number of healthy piglets,number of weak piglets,number of deformed piglets,number of stillborn piglets,and number of mummified piglets were recorded at day 14.The mouse study re-sults showed that muscle injection of pVAX1-SP-GHRH(1-44)plasmid followed by electrostim-ulation could promote mouse feeding and increase weight gain(P＜0.05),significantly increase mouse serum GHRH and IGF-1 levels(P＜0.05),and maintain its effects until day 21.The results of the pregnant sow study showed that the average birth weight of the piglets in the plasmid group was significantly increased(P＜0.01),and the placenta weight was significantly increased(P＜0.05).The serum GHRH and IGF-1 concentrations in the plasmid group sows were significantly increased(P＜0.01).The study results showed that muscle injection of pVAX1-SP-GHRH(1-44)expression plasmid followed by electrostimulation could promote mouse feeding and increase weight gain,and also significantly improve the average birth weight and placental weight of the piglets in pregnant sows.