Objective To investigate the value of tumor M2 pyruvate kinase (TuM2-PK ) ,cytokeratin-18 (CK18 )-3A9 and cytokeratin 19 fragment(CYFRA21-1) in non small cell lung cancer(NSCLC) ,and to evaluate the application value of combined de-tection of these three kinds tumor markers in diagnosis of NSCLC .Methods The serum levels of TuM2-PK and CK18-3A9 were measured in 67 patients with NSCLC(NSCLC group) ,72 patients with benign lung diseases(benign group) and 75 healthy control (control group) by enzyme linked immunosorbent assay(ELISA) .The level of CYFRA21-1 was detected by electrochemilumines-cence method .Results The serum levels of TuM2-PK ,CK18-3A9 and CYFRA21-1 in NSCLC group were higher than those in be-nign group and control group(P<0 .05) .The sensitivity of TuM2-PK or CK18-3A9 was better than CYFRA21-1(P<0 .05) ,the specificity of CYFRA21-1 was better than TuM2-PK or CK18-3A9(P<0 .01) .The sensitivity and accuracy of combined detection of TuM2-PK ,CK18-3A9 and CYFRA21-1 were both better than CYFRA21-1(P<0 .05) .The sensitivity ,specificity and accuracy of combined detection of TuM2-PK ,CK18-3A9 and CYFRA21-1 were all better than TuM2-PK or CK18-3A9 alone(P<0 .05) .Con-clusion The combined detection of TuM2-PK ,CK18-3A9 and CYFRA21-1 were better in sensitivity and accuracy of diagnosis of NSCLC .

Objective To explore the correlation of NT‐proBNP concentration and the fluid infusion volume in the patients with severe burn ,and to evaluate the significance of monitoring NT‐proBNP in severe burn patients .Methods Fifty cases of severe burn in 100 Hospital of PLA form September 2012 to September 2015 were selected ,without major disease history and genetic history before admission ,among them ,25 cases(group A) had the burned area 35% -50% TBSA or Ⅲ degree 10% -20% TBSA ,18 cases (group B) had the total burn area of >50% TBSA or Ⅲ degree >20% TBSA for group B and 10 cases(group C) had the total burn area>90% and were composite explosive injury .The NT‐proBNP concentrations were monitored on 1-30 d after admission for guiding blood transfusion and fluid infusion;contemporaneous age‐matched 53 patients with plastic surgery were selected as the control group .Compared with the calculation formula of the fluid volume ,the guiding advantage of NT‐proBNP was analyzed .Re‐sults The practical fluid volume in these burn patients were higher than that calculated by the common fluid replacement formula . The more severe burn ,the higher the NT‐proBNP concentration and the more blood products were demanded;the NT‐proBNP con‐centration ,fluid infusion volume ,MAP infusion volume and PLT transfusion volume had statistical difference between the group C and B and between the group B and A (P<0 .05) .Conclusion The NT‐proBNP concentration change may have a higher guidance and early warning significance for middle and long term rational control of blood transfusion and fluid infusion volume in the pa‐tients with severe burns .

Objective To investigate the serum levels of PGⅠ,PGⅡ,TK1,TSGF and CEA,CA724 in gastric cancer and eval-uate the application value of combined detection the above tumor markers in diagnosis of gastric cancer.Methods The serum levels of TSGF were measured in 94 patients with gastric cancer and 85 healthy control by rate method.PGⅠ,PGⅡ,TK1 and CEA,CA724 were detected by electrochemiluminescence method.Results PGⅠand PGⅠ/PGⅡwere lower than healthy control in serum of patients with gastric cancer (both P<0.05).There was no difference in PGⅡ (P>0.05),and other tumor markers were all higher than healthy control (all P<0.05).The sensitivity of PGⅠ,PGⅠ/PGⅡ were better than TK1,CEA and CA724 (all P<0.05),the specificity of PGⅠ/PGⅡ,CEA were better than TSGF (both P<0.05),the accuracy of PGⅠ,PGⅠ/PGⅡ were better than CA724 and TSGF alone (all P<0.05).When combined TSGF,TK1 and PGⅠ/PGⅡ,PGⅠ,the sensitivity was better than combined PGⅠ/PGⅡand PGⅠ alone (P<0.05).Then when added CEA, CA724,this sensitivity improved up to 82.98%.Although the combined detection would show a lower specificity,it still keep high to 84.71%.Combined detection improved the accuracy in diagnosis of gastric cancer,up to 83.80%.In this re-search,There was no difference in sensitivity,specificity and accuracy between the group of PGⅠ+PGⅠ/PGⅡ+TSGF+TK1+CEA and the group of PGⅠ+PGⅠ/PGⅡ+TSGF+TK1+CEA+CA724.Conclusion Compared with CEA and CA724 popular used in clinic,PGⅠ/PGⅡand PGⅠshowed a better application value.The group of PGⅠ+PGⅠ/PGⅡ+TSGF+TK1+CEA showed the best sensitivity.Combined detection of serum levels of PGⅠ,PGⅠ/PGⅡ,TSGF,TK1 ,CEA can significantly raise the sensitivity and accuracy in diagnosis of gastric cancer.

Objective To establish a liquichip method for detecting 6 sub-genotypes of hepatitis C virus(HCV),including 1a, 1b,2a,3a,3b and 6a.Methods The coupling method of PCR amplification and nucleic acid probe was established.The PCR product and the microspheres mixture of the coupled nucleic acid probe were hybridized for establishing the liquichip detection method.The sensitivity and specificity of the established liquichip detection method were evaluated.Nucleic acid in 93 serum samples was detec-ted by this method..Results The established HCV nuclei acid liquichip genotype detection method had the higher specificity and sensitivity,which could detect and classfy 6 HCV sub-genotypes.The sensitivity for HCV 1a,3a and 6a sub-genotypes was 1× 105 copies/PCR;the sensitivity for HCV 1b,2a and 3b sub-genotypes was 1×104 copies/PCR.The detection results in 93 serum samples showed that the this genotyping method had the characteristics of high throughput,rapidness,sentsitivity and specificity. Conclusion This method can be used for the simultaneous and quick detection of 6 HCV sub-genotypes and provides a new meth-od for the genotyping detection of HCV.

Objective To explore the diagnostic value of protein induced by vitamin K absence or antagonist-Ⅱ (PIVKA-Ⅱ) in non-infant with acquired deficiency of vitamin K-dependent coagulation factors (ADVKCF).Methods PIVKA-Ⅱ levels were measured by ELISA in 50 patients with ADVKCF on day 0,3,7 after vitamin K treatment.Prothrombin time(PT),APTT,FⅡ ∶ C,FⅦ∶ C,FⅨ∶ C,and FⅩ∶ C were analyzed simultaneously.Twenty healthy subjects were enrolled as controls.Results The average level of PIVKA-Ⅱ in ADVKCF group was (3.83 ± 1.40) μg/L,while (1.30 ± 0.54) μg/L in the control group (P ＜ 0.05).The PIVKA-Ⅱ levels on day 0 and 3 did not show significant difference [(3.83 ± 1.40) μg/Lvs (3.79 ± 0.66) μg/L,P ＞ 0.05],but decreasing significantly on day 7 compared to the control group (P ＜ 0.05).The PIVKA-Ⅱ level was (3.78 ± 1.30) μg/L in patients receiving plasma transfusion,while (3.91 ± 1.49)μg/L in no-plasma-transfusion group (P ＞ 0.05).Coagulation factors Ⅱ,Ⅶ,Ⅸ and Ⅹ activity which decreased significantly before treatment returned to normal range after one week use of vitamin K,leading to complete correction of prolonged APTT and PT (＞ 100 seconds).Conclusions The PIVKAⅡ level in ADVKCF patients is significantly higher than that of healthy subjects within one week treatment of vitamin K,which is not influenced by plasma transfusion.This study suggests that PIVKA-Ⅱ is a more sensitive parameter than APTT,PT and the activity of coagulation factor,which could be a valuable factor in the early diagnosis of ADVKCF.

Objective:To investigate the content and mode of pharmaceutical care for the patients with severe burn and promote the rational use of drugs. Methods:Taking the treatment for one patient with severe burn as example,pharmacists provided pharmaceu-tical care in respects of anti-infection therapy,organ preservation,nutritional support, drug interactions, drug precautions, drug incom-patibility, adverse drug reactions and the effect evaluation . Results:The program of rational drug use was provided for clinics through the implementation of pharmaceutical care. The vital signs of the patient were stable, and then the patient left on pass and continued to be treated with rehabilitation therapy. Conclusion:It is necessary for clinical pharmacists to perform pharmaceutical care for the pa-tients with severe burn,reduce the abuse of drugs and improve the medication safety and effectiveness. Meanwhile, the collaborative service of doctors,pharmacists,nurses and inspectors to patients reflects the value of clinical pharmacists.

Objective To investigate the expression of Circulating tumor cells ( CTCs ) in peripheral blood cells of patients with different stages of Small-Cell Lung Cancer ( SCLC) ,and to evaluate its significance in early diagnosis of lung cancer metastasis.Methods Thirty-five patients with SCLC ( 12 in limited stage and 23 in extensive stage ) and 25 patients with benign lung disease were recruited at the Shanghai Changhai Hospital from April 2011 to April 2013.Samples were prepared from 7.5 ml peripheral venous blood and collected in CellSave tubes.The CTC counts were determined using CellTracks AutoPrep fluorescence scanning system according to the manufacturers′instructions.The expression of CTCs in peripheral blood of SCLC patients and benign lung disease patients were analyzed.The expression of CTCs in different stages of SCLC was measured and compared.Furthermore, samples were prepared from 2 ml peripheral venous blood by centrifugation.The serum levels of NSE Neuron specific enolase were measured.The relationship between the expression of CTC and NSE was analyzed.Results CTCs positive rates in SCLC were significantly higher than in benign lung disease controls [ Positive rates of CTC≥1 were 80.0%and 12.0%(χ2 =27.003,P<0.000 1),CTC≥5 were 51.4%and 0 (χ2 =18.367,P<0.000 1), CTC≥10 were 34.3%and 0(χ2 =10.714,P<0.001),CTC≥50 were 17.1 and 0(P=0.036,P<0.05), respectively ].CTCs positive rates in SCLC extensive stage were significantly higher than limited-stage [Positive rates of CTC≥1 were 58.3% and 91.3%(P=0.033,P<0.05), CTC≥5 were 65.2% and 25.0%(P=0.035,P<0.05), respectively].The expression of CTCs was significantly correlated with NSE (r=0.743 7, P<0.000 1).Conclusion The expression of CTC in SCLC patients is significantly higher than those in control group , and is closely related to clinical stages , which may provide new clues to early predicting of SCLC metastasis and deterioration.

OBJECTIVETo investigate the expression of IL- 23/IL- 17 axis in peripheral blood of patients with primary immune thrombocytopenia （ITP） and its clinical significance.

METHODSThe real-time quantitative reverse transcription-polymerase chain reaction（RT-PCR）was used to determine the expression of IL-23p19, p40, p35, IL-23R, IL-12Rβ1, IL-12Rβ2, IL-17A, IL-17F mRNA in the peripheral blood of 45 ITP patients and 30 healthy controls. The correlations between the expression of IL-23 and IL- 17, platelet counts, serum cytokine concentrations of ITP patients were analyzed. Furthermore, nine newly diagnosed ITP patients were followed up during treatment.

RESULTSThe gene expressions of IL-23p19, p40, IL-23R, IL-12Rβ1, IL-17A, IL-17F in ITP patients were significantly higher than those in healthy controls, the relative expression levels of ITP were 5.58, 2.13, 4.20, 2.45, 4.29, 2.50 times as much as that of healthy controls. And elevated serum IL-23［（198.70±94.56）ng/L vs（50.72±22.97）ng/L, t= 10.06, P<0.001］, IL-17［（85.25±21.97）ng/L vs（11.39±4.27）ng/L, t=21.94，P<0.001］levels were also observed in these ITP patients. In addition, the serum IL-23 level in ITP patients was positively correlated with IL-17（r=0.496, P<0.01）, but negatively correlated with the platelet counts（r=-0.408, P<0.01）, and IL-17 level was negatively correlated with platelet counts（r=-0.464, P<0.01）.

CONCLUSIONThe IL-23/IL- 17 expression in ITP patients was significantly elevated, indicating IL-23/IL-17 play an important role in the pathogenesis of ITP.

Case-Control Studies ; Cytokines ; blood ; Gene Expression ; Humans ; Interleukin-17 ; metabolism ; Interleukin-23 ; metabolism ; Platelet Count ; Purpura, Thrombocytopenic, Idiopathic ; metabolism ; RNA, Messenger

OBJECTIVETo study the efficacy and safety of rituximab (RTX) in the treatment of idiopathic thrombotic thrombocytopenic purpura (ITTP).

METHODSAmong 17 ITTP patients, nine cases of the RTX group were administrated with RTX plus plasma exchange (PEX) and steroids. Eight cases of the control group received PEX plus steroids±other immune inhibitors. Patients received RTX 375 mg/m², 1 per week for 4 weeks. The laboratory parameters, including hemogram, LDH, ADAMTS13 activities and its inhibitors, and the ratio of B lymphocytes in peripheral blood were monitored. The number of PEX, total plasma volumes, remission time, relapse ratio and adverse effects in both groups were compared.

RESULTSThe median number of PEX/median total plasma volumes in the RTX and control group were 5(2-8)/9.6(4.0-15.4) L and 6(4-9)/11.2(7.5-14.6) L, respectively. Patients in the RTX and control group achieved hematologic remission at the median time of 15(5-20) days and 22(7-36) days, respectively. And the median time of immunological remission in the two groups was 2(2-8) and 2(2-4) weeks, respectively. ADAMTS13 activities increased significantly after 2 weeks in both two groups. There was no relapse in the RTX group, while 4 patients relapsed in the control group. The percentage of B lymphocytes in peripheral blood obviously deduced one week after first dose of RTX infusion compared with the level before treatment [(2.19±5.11)% vs (18.39±7.15)%, P<0.001], and began to gradually increase 9 months later. Severe adverse events were not observed in RTX group during the therapeutic procedure and follow-up, but one patient, who had sustained immunologic remission, died of severe pneumonia 7 months later.

CONCLUSIONIn the treatment of ITTP, RTX in conjunction with PEX and steroids appeared to be a safe and effective therapy, with fast and sustained remission in hematology and even in immunology, with lower relapse rate and less adverse effects. But patients needed to be paid attention to prevent and treat infectious events in time.

ADAM Proteins ; ADAMTS13 Protein ; B-Lymphocytes ; Humans ; Plasma Exchange ; Purpura, Thrombotic Thrombocytopenic ; Recurrence ; Rituximab ; Steroids