Objective To test the changes of lymphocyte subsets and anti-GP Ⅱ b/Ⅲa,anti-GP Ⅰ b/Ⅸ specific antibodies in patients with idiopathic thrombocytopenic purpura(ITP),and discuss the related factors involved in pathogenesis of ITP.Methods The changes of lymphocyte subsets CD3+,CD4+,CD8+,CD4+CD8+,CD19+ in 72 patients with ITP (ITP group) and 50 healthy controls (control group) were measured by flow cytometry (FCM).Expression of anti-GP Ⅱ b/Ⅲ a,anti-GP Ⅰ b/Ⅸ specific antibodies were tested by monoclonal antibody immobilization of platelet antigens assay (modified MAIPA).Results Optical density of anti-GP Ⅱ b/Ⅲ a,anti-GP Ⅰ b/Ⅸ specific antibodies in ITP group was much higher than that in control group (P ＜0.05).In changes of lymphocyte subsets,the number of the CD3+,CD4+,CD4+/CD8+ in ITP group were obviously lower than those in control group (P ＜0.01),but CD8+ and CD19+ were much higher than those in control group(P ＜0.01).Conclusions Expression of lymphocyte subsets and platelet specific antibodies can reflect the pathogenesis of ITP well.The abnormal function and ratio of lymphocyte subsets,as well as the immunologic mechanism disorder of lymphocytes play important roles in the pathogenesis of ITP.

Objective To analyze the clinical features of eosinophilic granulomatosis with polyangiitis (EGPA).Methods The clinical characteristics,laboratory test,therapeutic regimen,pathology and imaging diagnosis in 23 cases with EGPA from the First Affiliated Changhai Hospital of Second Military Medical University within June 2007 to January 2013 were retrospectively analyzed.Results There were 14 males and 9 females,with the age ranged from 21 to 68 years.The mean age was (42±9) years.The overall prevalence of asthma was 78％(18/23).The allergic rhinitis accounted for 61％(14/23).The maxillary sinuses were the most frequendy involved,which accounted for 57％(13/23).Skin involvement was 57％(13/23),peripheral neuritis was 70％(16/23).Central nervous system involvement presented cerebrovascular event.Cardiac involvement accounted for 48％(11/23),digestive system involvement accounted for 17％(4/23).The outcome of auxiliary laboratory test revealed that 96％(22/23) patients expressed significantly higher levels of IgE and 70％(16/23) patients carried anti-neutrophil cytoplasmic antibodies (ANCA) which were presented as the perinuclear ANCA (p-ANCA).The patchy infiltrates of lung CT scan accounted for 69％ (11/23).EMG showed mononeuritis multiplex and symmetric sensory motor neuropathy.The abnormal ratios of ECG and Echocardiography were 48％(11/23),79％(11/14) respectively.The pathological manifestations were necrotizing vasculitis,eosinophilic tissue infiltration,and extravascular granulomas.Conclusion Our results confirm the heterogenicity in clinical presentation and lack of specificity.Early diagnosis and treatment will be helpful for good prognosis.

Objective To investigate the effect of different temperature moxibustion on serum IL-1β, IL-2 and TNF-αcontents in acute adjuvant arthritis rats and provide a basis for the mechanism of local anti-inflammatory and immune action of moxibustion. Methods A rat model of adjuvant arthritis (AA) was made by Freund’s complete adjuvant (FCA) modeling method. Of 32 SD rats, 8 were randomly selected as a normal group and the other 24 for model making. After successful model making, the rats were randomly allocated to model, treatment 1 and treatment 2 groups, 8 rats each. Treatment 1 and 2 groups received moxibustion. Local temperature at moxibustion point was controlled at (38±1)℃in treatment 1 group and at (45±1)℃in treatment 2 group. Rat serum IL-1β, IL-2 and TNF-αcontents were measured by enzyme-linked immunosorbent assay (ELISA). Results There were statistically significant differences in serum IL-1β, IL-2 and TNF-αcontents between the model, treatment 1 or treatment 2 group and the normal group (P<0.01). There was a statistically significant difference in serum IL-2 content between treatment 1 and the model groups (P<0.01). There were statistically significant differences in serum IL-1β, IL-2 and TNF-αcontents between treatment 2 group and the model or treatment 1 group (P<0.01). Conclusions Moxibustion has a reducing effect on serum IL-1βand TNF-αcontents and a raising effect on IL-2 content. 45℃moxibustion temperature can improve the effect of moxi bustion. The anti-inflammatory action of moxibustion may be through the mechanism of reducing serum IL-1βand TNF-αcontents and raising IL-2 content, which relieve body inflammatory reaction. The action of moxibustion needs proper moxibustion temperature.

Objective According to the level of serum cystatin C (Cys-C)in patients with multiple myeloma (MM),to explore its significance of early assessment to renal damage.Methods To select 42 patients with MM(MM group) and 30 healthy subjects(control group),and measure serum Cys-C,creatinine (SCr) and β2-microglobulin (β2-MG).Results The level of serum Cys-C in MM group was significantly higher than that in control group [(2.34 ± 0.57) mg/L vs.(0.66 ± 0.12) mg/L],which increased with ISS stage[Ⅰ,Ⅱ,Ⅲ stage was(1.77 ± 0.38),(2.11 ± 0.46),(3.14-± 1.07) mg/L](P＜ 0.05).Cys-C and β 2-MG,renal function had a positive correlation (r =0.760,0.694,P ＜ 0.05).Conclusion The increasing of Cys-C in MM patients is related to impaired renal function and tumor burden,which can be used as an early assessment indicators.

Colorectal Cancer (CRC) is a kind of molecular diversified neoplasm.For nonmetastatic early CRC patients who received surgery based systemic treatment,the 5 years survival rate is as high as 50％-80％.However,only chemotherapy is available for majority of metastatic advanced CRC patients.The effect of chemotherapy is disappointing.Numerous studies have demonstrated that some patients with metastatic advanced CRC can benefit from anti-EGFR and anti-VEGF monoclonal antibodies.Here,we give a brief overview about the clinical research of the detection of KRAS,BRAF,MSI and HER2in guiding treatment with targeted drugs in metastatic advanced CRC

AIM:To study the effects of ischemic postconditioning on cerebral ischemia following middle cerebral artery occlusion in rats. METHODS: 21 rats were randomly divided into three groups: middle cerebral artery occlusion (MCAO), MCAO+transient unilateral common carotid artery occlusion (u-CCA-O), MCAO+transient bilateral common carotid artery occlusion (b-CCA-O)(n=7, respectively). u-CCA-O/b- CCA-O was generated by transient middle cerebral artery occlusion plus transient unilateral/bilateral common carotid artery (CCA) occlusion. After the suture was removed, ischemic postconditioning was performed by occluding CCA for 10s, reperfusion 10s, and then allowing for another 4 cycles of 10s of reperfusion and 10s of CCA occlusion. Rats were sacrificed 2 d later and infarct size was measured. Cerebral blood flow (CBF) was measured in different 15 time points: 0 min, 10 min, 1 h after MCA occlusion, 0 min after MCA reperfusion, 10s of CCA occlusion and 10s of CCA reperfusion in all five cycles, 30 min after MCA reperfusion. Functional neurological outcome was determined 1 h and 48 h after reperfusion. Infarct volume was measured 48 h after reperfusion. RESULTS: The infarct volumes in u-CCA-O group and b-CCA-O group diminished compared to the control group. The results of CBF demonstrated that b-CCA-O group diminished 9% compared with control and u-CCA-O group when 30 min after intervention. The rats in u-CCA-O and b-CCA-O group had better neurological performance at 1 h after reperfusion. CONCLUSION: Ischemic postconditioning reduces infarct size, improves functional neurological outcome, most plausibly by diminishing cerebral blood flow.

Objective To investigate effects of Danhong on the serum levels of CD137, high-sensitivity C-reactive protein (hs-CRP) and homocysteine (Hcy) in patients with non-ST elevation acute myocardial infarction complicating metabolic syndrome. Methods A total of 126 patients with non-ST elevation acute myocardial infarction complicating metabolic syndrome were enrolled and randomly divided into a conventional treatment group and a Danhong treatment group using a random-digit table, with 63 patients in each group. All patients underwent angiography or percutaneous coronary intervention. The patients in the Danhong treatment group treated with intravenous Danhong 20 ml on the basis of conventional treatment for 1 week. The serum levels of CD137, hs-CRP and Hcy were measured at hospital admission and 10 days after treatment. The severity of coronary artery disease was assessed by the Gensini-score. Results The levels of CD137, hs-CRP and Hcy in both groups after treatment were significantly lower than before treatment (conventional treatment group: t 12.393, 17.408 and 9.458; Danhong treatment group: t 16.110, 17.573 and 13.481; all P<0.01), and the Danhong treatment group were significantly decreased than the conventional treatment group (t 2.815, 3.224 and 3.157, all P<0.01). The serum levels of CD137 and hs-CRP before treatment were significantly correlated with Gensini scores in 126 patients (r 0.720 and 0.562,all P<0.01). Conclusions The serum levels of CD137 and hs-CRP are significantly correlated with the severity of coronary artery disease, intravenous Danhong may has protective effect for coronary artery disease via decreasing CD137 and hs-CRP.

This study sought to construct recombinant eukaryotic plasmid pcDNA3. 1-sFlt-1 and observe its effect on proliferation of vascular endothelial cells. Total RNA was extracted from human umbilical vein endothelial cells (HUVECs) firstly. The 1st-3rd Ig-like domains of Flt were amplified by polymerase chain reaction (PCR) from the full-length cDNA. Subsequently, the PCR product was cloned into the eukaryotic plasmid pcDNA3.l(+)/myc-His. The constructed recombinant plasmid pcDNA3. 1-sFlt-1 was sequenced. Then recombinant plasmid was transfected into Lewis lung cancer cells. RT-PCR and SDS-PAGE were used to detect the expression of soluble vascular endothelial growth factor (VEGF) receptor mRNA and protein, respectively. MTT method was used to evaluate the effect of sFlt-1 protein on proliferation of HUVECs induced by VEGF. The results showed: (1) The sequence of inserted fragment was correct. (2) Lewis lung cancer cells with recombinant plasmid transfection could express the soluble VEGF receptor mRNA and protein stably. (3) Culture supernatant of Lewis lung cancer cells with sFlt-1 could significantly inhibit the proliferation of HUVECs induced by VEGF. These data suggested that recombinant eukaryotic plasmids pcDNA3. 1-sFlt-1 was constructed successfully, sFlt-1 mRNA and protein were expressed in eukaryotic system stably and sFlt-1 protein could significantly inhibit the proliferaton of endothelial cells induced by VEGF.
Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cells, Cultured ; Genetic Vectors ; genetics ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Transfection ; Vascular Endothelial Growth Factor A ; pharmacology ; Vascular Endothelial Growth Factor Receptor-1 ; biosynthesis ; genetics ; pharmacology

Objective:To observethe influence of electroacupuncture (EA) on histomorphologies of lacrimal glands, cornea and conjunctiva in experimental dry eye syndrome, and to explore the repair effects of EA on lacrimal glands and ocular surface damage. Methods:Twenty-four healthy male New Zealand rabbits were randomly divided into a normal group, a model group, an EA group and a medication group, 6 rabbits in each group. Experimental dry eye syndrome models were prepared in rabbits by using 0.1% benzalkonium chloride for eye drops. Tear secretion volume, break-up time of tear film (BUT) and corneal fluorescein staining score were observed before and after the treatment. Periodic acid Schiff (PAS) staining method was used to observe the changes of conjunctival goblet cells in rabbits. After hematoxylin eosin (HE) staining, morphological changes of rabbit cornea, conjunctiva and lacrimal gland tissues were observed under light microscope. Results: Compared with the normal group, tear secretion volume and BUT were significantly reduced (bothP<0.01), while the corneal fluorescein staining score was significantly increased (P<0.01) in the model group. Compared with the model group, tear secretion volume and BUT were significantly increased, while the corneal fluorescein staining score was significantly decreased in the EA group and the medication group (allP<0.01). Compared with the normal group, the number of conjunctival goblet cells in the model group was significantly reduced; compared with the model group, the numbers of conjunctival goblet cells were all relatively higher in the EA group and the medication group. Pathological lesions of cornea, conjunctiva and lacrimal glands all showed improvement by HE staining in the EA group and the medication group after the intervention. Conclusion:EA can improve tear secretion and tear film stability of rabbit dry eye syndrome, and repair the pathologic lesions of conjunctival goblet cells, corneal epithelia, cornea, conjunctiva and lacrimal glands.

Objective To investigate the clinical significance of Epstein-Barr virus EBV DNA in children with Epstein-Barr virus infection realated diseases.Methods A retrospective cohort study was performed.Totally 222 blood samples were collected from children who were diagnosed as EBV infection in Shandong Provincial Hospital from June 2012 to August 2013.Fluorescent quantitative PCR( FQ-PCR) was used to analyze the EBV DNA in peripheral blood lymphocytes.ELISA was used to analyze the four EBV serology antibodies in the serum.Two groups of tested results were compared.Heart, hepatic impairment and renal function were analyzed through detecting AST, ALT, BUN, CREA, CK, CKMB.The results were grouped by EBV DNA copy number, and then non-parametric test together with correlation analysis was performed using SPSS21.0 analytics software.Results The positive rate of EBV-CA IgM and EBV DNA was 51.35%(114/222) and 72.97% (162/222) respectively, χ2 =24.01, P<0.001.The EBV DNA copy number was significant difference (χ2 =11.79,P<0.05) in children at different stages of infection ( no previous infection:6.30 ×103 -1.20 ×104 copies/ml;early infection:5.56 ×103 -3.92 ×106 copies/ml;acute infection:6.58 ×103 -1.73 ×106 copies /ml;chronic infection or recurrent infection:8.92 ×103 -2.34 ×104 copies/ml;late infection or recovery:5.20 ×103 -1.12 ×107 copies/ml;past infection:5.46 × 103 -1.33 ×104 copies/ml).Each biochemical targets were divided into five groups by EBV DNA copy number (Ⅰ>1 ×106 copies/ml,Ⅱ1 ×105 -1 ×106 copies/ml, Ⅲ1 ×104 -1 ×105 copies/ml, Ⅳ5 × 103 -1 ×104 copies/ml, Ⅴ<5 ×103 copies/ml), and ALT(χ2 =10.14,P<0.05), BUN(χ2 =18.17, P<0.05), CK(χ2 =13.09,P<0.05), CKMB(χ2 =17.93,P<0.01) had a statistically significant difference between each group.Well, the log value of EBV DNA copy number had a positive correlation relationship with AST(r=0.357,P=0.001), ALT(r=0.376,P=0.001), BUN(r=0.329,P=0.000), CK(r=0.235,P=0.035).Conclusions Detection of EBV DNA can be used for the early diagnosis and assessment of process of the EBV infection related disease in children.The detection of liver, kidney function and myocardial enzymes can be used for evaluating the severity of EBV infection.