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【Objective】 To construct an in-vitro model of erythrocyte antibody-mediated complement activation, and establish quantitative detection methods based on flow cytometry and spectrophotometry, so as to explore the correlation of anti-body titers and complement activation speed, and provide a methodological basis for studying the adverse transfusion reactions of anti-body mediated complement hemolysis. 【Methods】 Mouse monoclonal antibody that recognized human C3b and fluorescent secondary antibody were used to label C3b fragments on erythrocytes, and the deposition of C3b fragments after complement activation was detected by flow cytometry. The absorbance at 540 nm of the supernatant in the complement activation reaction system was measured by spectrophotometry as the amount of hemoglobin released was related to the absorbance. 【Results】 The complement activation system was constructed according to the ratio of 3% red blood cell suspension (mixed for 6 people) 1∶anti-Tja 1∶complement 2. The repeatability was good (P value>0.05) as different red blood cell mixtures had been used to repeat the detection reaction system. When using 32×, 64× and 128× dilutions of anti-Tja mediated complement activation, the deposition of C3b fragments has been detected by flow cytometry at 30 s, 1 min and 2 min, respectively, and MFI peaked at 5 min, 10 min and 30 min, respectively. No obvious hemolysis has been observed within 1.5 h. 【Conclusion】 In vitro model of anti-Tja-mediated complement activation demonstrates the speed of complement activation is related to the concentration of antibody. At a certain antibody concentration, the speed of complement activation has been slowed down, and no obvious hemolysis observed.
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【Objective】 To understand the distribution and gene frequency of main red blood cell blood groups in Lahu ethnic minority and analyze the genetic characteristics of Lahu people. 【Methods】 1) ABO forward and reverse typing had been performed by microplate method; 2) Rh, MN, H, P1Pk and Mur antigen were tested by the tube method. If the ABO forward and reverse typing were incompatible, the tube method was used for confirmation. 【Results】 The distribution characteristics of blood group and gene frequency in Lahu ethnic minority were as follows: B>O>A>AB for ABO, with genotype frequency as p 11.1%, q 27.5% and r 61.4%; the frequency of Rh genotype was CDe 83.3%, cDE 12.0%, cDe 2.42%, CDE 2.32%, CdE 0%, Cde 0%, cdE 0% and cde 0%; M > MN>N for MN blood group, with genotype frequency as M 75.26% and N 24.74%; P1
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【Objective】 To develop a novel screening reagent for -D- phenotype preliminary screening based on the difference in RhD antigen expression level of -D- phenotype and normal RhD phenotype. 【Methods】 RhD antigen expression of -D-phenotype and Rh D-- gene carrier were detected by flow cytometry. By adjusting the concentration of polybrene in the screening system, the red blood cells with high RhD antigen expression level agglutinated, and the preliminary screening of the -D-phenotype and its gene carriers was realized. 【Results】 According to the quantitative results of immunofluorescence intensity (MFI) analysis by flow cytometry, the expression level of RhD antigen in -D- phenotype cells (284 360±16 698, n=3) was about 3 times normal RhD positive cells (98 642±35 908, n=9)(P<0.01), while RhD antigen expression level of RhD-- gene carrier (181 109±39 455, n=4) was about 2 times normal RhD positive cells(P<0.01). RhD antigen expression (144 538±227 445, n=7) of the positive cells screened by 15 μL 3% fresh red blood cell suspension and screening system 35 μL (1 μL IgG anti-D, 29 μL polybrene polybrene, and 5 μL low ionic strength solution) was about 1.5 times normal RhD positive cells. 【Conclusion】 The polybrene preliminary screening system, which can be used for high-throughput screening of -D- phenotype, is a reliable technical method for frequency study of this phenotype.
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Objective With the purpose of further improving blood safety and providing the reference basis for a strategy suitable for large-scale routine screening of irregular antibodies in blood service,the irregular antibodies in blood samples from donors in Shanghai area were investigated.Method Irregular blood antibodies were screened in 824 072 blood donors through combined method of saline medium and microplate papain.Result 1 246 samples of blood donors were detected with irregular antibodies among the total 824 072 samples which indicate the positive rate of 1.51‰.Conclusion Screening of irregular antibodies in blood samples from donors can effectively reduce or avoid hemolytic transfusion reactions.The combined method of saline medium and microplate papain is an effective method for large-scale routine screening of irregular antibodies in blood service.
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Objective To study the methods andfeasibility of re-entry of NAT reactive blood donors.Methods ELISA Non-Reactive(NR) and NAT Reactive(R) Blood Donors in 2012-2014 were selected and informed consent.Blood samples collected were tested:ELISA (HBsAg、Anti-HCV、Anti-HIV and Anti-TP) and NAT (HBV-DNA、HCV-RNA and HIVRNA).ELISA NR/NAT NR samples were tested by ECLA for HBsAg、HBeAg、Anti-HBs、Anti-HBc、Anti-HBe、Anti-HCV、Anti-HIV.Results 30 donors were called back,and 11 of them were ELISA NR/NAT R,which were deferred permanently.19 of them were ELISA NR/NAT NR,and the ECLA results were 89.5％ ELISA NR/NAT NR donors were Anti-HBc R and/or Anti-HBe R (12 cases Anti-HBc/Anti-HBe R、5 cases Anti-HBe R).Anti-HBc R or Anti-HBe R donors had risk of infection,which were deferred permanently.2 cases all items Non-Reactive may be eligible for re-entry.Conclusion Re-entry of NAT reactive blood donors is very necessary and meaningful,but in the view of blood safety,it is important that establish a set of safe program for Chinese.
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Objective To research the clinical outcome of atypical glandular cell (AGC)according to various subtypes of HPV infection and histological pathology results.Methods The data of the liquid-bases cytology (LBP),HPV infection and histology in 102 cases of AGC at the gynecology outpatient department of our hospital from January 1 ,2009 to February 28,2014 were collected and performed the analysis on their clinical outcomes.Results Among 67 218 cases of LBP detection,102 cases were AGC with the total incidence rate of 0.15%.In the cases of AGC-NOS,67 cases were normal or benign lesions,11 cases were precancerous lesion and malignant lesions;in the cases of AGC treading to tumor,the benign,precancerous and malignant lesions were in 7,14 and 3 ca-ses respectively.At the same time in the cases of AGC-NOS,HPV infection was in 64 cases,in which 57 cases were high risk infec-tion(type 16,52,45)and 7 cases were low risk infection(type 6,11 ).The single infection,double infection and multiple infection were in 54,6 cases respectively;in the cases of AGC trends to neoplasm,HPV infection was in 19 cases,in which 18 cases were high risk infection(type 52,16,18)and 1 case was low risk infection(type 6),single infection and double infection were in 15 cases and 4 cases respectively.Conclusion AGC may play an important role for the forecast of cervical malignant lesions.The results of differ-ent HPV subtypes infection in AGC related tumors also play a certain role in the prediction of cervical neoplasia.Their combined a-nalysis is the important signal for judging the occurrence of gynecological cervical precancerous lesion and malignant tumor,i.e., AGC combining with the corresponding HPV subtype infection not only can make a judgement for the cervical lesions,but also pro-vides the basis for predicting the high risk existence of gynecological malignant tumor and provides constructive suggestions for Chi-na regional cervical carcinoma vaccine manufacturing and promotion.
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Objective To investigate the blood coagulation function changes and intervention effects of metformin in patients with impaired glucose tolerance.Methods One hundred patients with impaired glucose tolerance and 50 healthy volunteers (control group) were recruited.The patients with impaired glucose tolerance were divided into test group 1 and test group 2 by random digits table method.The patients in test group 2 were given metformin and the patients in test group 1 were given the same dosage of placebo.The oral glucose tolerance test (OGTT),glycosylated hemoglobin (HbA1c),prothrombin time (PT),activated partial thromboplastin time (APTT),thrombin time (TT) and fibrinogen (Fib) level were determined before and after treatment.Results Compared with those in control group,the patients in test group 1 and test group 2 had higher level of 2 h postprandial blood glucose (2 h PBG),HbA1c and Fib,and lower level of APTT,PT (P ＜ 0.05).The level of 2 h PBG,HbA1c and Fib in test group 2 were significantly lower than those in test group 1,(6.6 ± 2.5) mmol/L vs.(7.9 ± 3.6) mmol/L,(5.2 ± 1.3)％ vs.(5.8 ± 1.4)％,(3.0 ± 1.0) g/L vs.(3.6 ± 1.2) g/L,and the level of APTT and PT were significantly higher than those in test group 1,(34.3 ± 6.8) s vs.(31.6 ± 6.5) s,(12.6 ± 0.7) s vs.(12.3 ± 0.7) s,P ＜ 0.05.Compared with those in control group,there were no significant differences (P ＞ 0.05).Pearson line correlation analysis revealed that APTT and PT had negative correlation with 2 h PBG and HbA1c (P ＜ 0.01),and Fib had positive correlation with 2 h PBG and HbA1c (P ＜ 0.01).TT had no correlation with 2 h PBG and HbA1c (P ＞ 0.05).Conclusion Blood coagulation function of patients with impaired glucose tolerance may have some alteration,and appropriate therapeutic intervention can reduce the risk factors for cardiovascular disease.
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Objective To analyze clinical features of and hypertrophy-related factors in patients with hypertrophic port-wine stains (PWS). Methods Patients with PWS were enrolled into this study from Anhui Provincial Hospital and the First Affiliated Hospital of Anhui Medical University between January 2010 and August 2014. Clinical features of hypertrophic PWS were investigated. The factors related to hypertrophy in PWS were analyzed by univariate and multivariate unconditional logistic regression analyses. Results A total of 262 patients with PWS were enrolled, 82 (30 males and 52 females)of whom had hypertrophic PWS with a median age of 32.5 years (range, 18 - 54 years). Among the 82 patients, 66(80.48%)had plaque-like hypertrophic PWS, 9(10.98%)had papular or nodular type, and 7 (8.54%)had mixed type; 56.10% (46/82)were aged ≥ 30 years, 41.46% (34/82)varied from 11 to 30 cm2 in lesional area, and 85.36% (70/82)showed purple lesions. There was a significant difference between patients with hypertrophic PWS and those with flat PWS in the distribution of age, lesional area and color(χ2 = 25.559, 10.580, 90.630, respectively, all P 0.05). Multivariate unconditional logistic regression analysis revealed that an age ≥ 30 years(OR = 2.889, 95%CI: 1.459 - 5.721)and purple lesions(OR = 19.984, 95% CI: 5.704 - 70.023)were factors related to skin hypertrophy in PWS. Conclusion An age ≥ 30 years and purple lesions seem to be hypertrophy-related factors in PWS.
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Objective To investigate the expression of heat shock protein 5 (HSPA5) in breast carcinoma and to study its relationship with clinical and pathological factors.Methods HSPA5 from 97 cases of breast carcinoma and 40 cases of adjacent tissues of breast carcinoma were examined by immunohistochemical method.Results The overall positive rate of HSPA5 in 97 cases of breast cancer was 81.4 ％ (79/97).The positive rates of HSPA5 in cancer tissues and paired normal tissues were 32.5 ％ (13/40) and 7.5 ％ (3/40),respectively.The expression rate of HSPA5 in tumor tissues was significantly higher than that in normal tissues (x2 =7.813,P =0.038).The expression of HSPA5 in breast carcinoma was correlated with histologic grade and lymph node metastasis (x2 =22.233,P =0.007; x2 =5.342,P =0.024).Conclusion The occurrence of breast carcinoma and the formation of invasion may be related with overexperssion of HSPA5.
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Objective To study the effect of different grading glycosylated hemoglobin(HbA1c) on carotid artery endothelial cells in diabetic patients.Methods A total of 120 patients with type 2 diabetes mellitus patients were divided into 4 groups according to the different levels of HbA1c:6％ ≤HbA1c ＜ 7％ as group 1 (30 cases),7％ ≤HbA1c ＜ 8％ as group 2(31 cases),8％ ≤HbA1c ＜ 9％ as group 3(29 cases) and HbA1c ≥ 9％ as group 4 (30 cases).The indicators of endothelial function and carotid ultrasonography were detected and recorded.Results There was significant difference in endothelin-1 (ET-1) and nitric oxide (NO) among 4 groups (P ＜ 0.01).When compared between 2 groups,ET-1 and NO in group 4 was the highest [(89.3 ± 19.2) ng/L,(59.5 ± 9.3) μ mol/L],next was group 3 [(85.9 ± 16.4) ng/L,(56.4 ± 8.9)μ mol/L] and group 2[(83.4 ± 17.3) ng/L,(52.3 ± 8.3) μ mol/L],and there was significant difference (P ＜0.05).There was significant difference in carotid intima-mild thickness (IMT) among 4 groups (P ＜ 0.01).When compared between two groups,IMT in group 4 was the biggest [(2.90 ± 0.62) mm],next was group 3 [(2.40 ± 0.73)mm] and group 2[(1.90 ± 0.42) mm],and there was significant difference(P ＜ 0.05).There was significant difference in the incidence of carotid plaque among 4 groups (P ＜ 0.01).When compared between 2 groups,the incidence of carotid plaque in group 4 was the highest [93.3％(28/30)],next was group 3 [79.3％(23/29)]and group 2 [61.3％ (19/31)],and there was significant difference(P ＜ 0.05).There was significant difference in the type of carotid plaque among 4 groups (2 =51.10,P ＜ 0.01).HbA1c was correlated with the type of carotid plaque (r =0.56,P ＜0.01).Conclusion For patients with type 2 diabetes,there is a strong link between plaque,HbA1c level and carotid artery endothelial cells.
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Objective To investigate the expression of MUC-1 and CD44v6 in breast carcinoma and its diagnostic value.Methods 93 cases of breast carcinoma were examined by immunohistochemical method for MUC-1 and CD44v6.Results The positive rates of MUC-1 and CD44v6 were 71.0 ％ (66/93) and 63.4 ％ (59/93),respectively.The expression of MUC-1 in breast carcinoma was correlated with histologic differentiation (14.3 ％,69.4 ％,100.0 ％) and lymph node metastasis (22.9 ％,100.0 ％) (x2 =36.147,63.047,both P ＜ 0.0001),but had no relationship with the type of breast carcinoma,tumor clinical stages,and tumor size (all P ＞ 0.05).The expression of CD44v6 in breast carcinoma was correlated with tumor clinical stages (6.3 ％,64.6 ％,93.1％) and lymphnode metastasis (20.0 ％,89.7 ％) (x2 =9.507,45.662,both P＜0.05),but had no relationship with tumor type,tumor clinical stages and tumor size (all P ＞ 0.05).Conclusion MUC-1 and CD44v6 may play important roles in the development of breast carcinoma and the biologic behavior of breast carcinoma may be closely related with over expression of MUC-1 and CD44v6 protein.MUC-1 and CD44v6 can be used to predict the prognosis and direct the treatment of breast carcinoma.
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Morphological examination of blood cells is an important part of the hematology examination course. In order to enrich teaching resources, network of bilingual education resource was established and put into application by the Department of Hematology in Guangzhou Medical College. The repository improved teaching quality of cell morphology, and played a role in training personnel of hematology examination with solid basic skills.
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Objective To analyze the clinic features and hereditary characteristics of three subtypes of porokeratosis, namely disseminated superficial actinic porokeratosis (DSAP), porokeratosis palmaris et plantaris disseminata(PPPD) and porokeratosis of Mibelli (PM) in five pedigrees with porokeratosis. Meth-ods After clinical and pathological diagnosis, every living family member of the five pedigrees with poro-kerotosis was undergoing medical examination and genetics analysis. These five pedigrees consisted of three DSAP pedigrees (totally 266 family members including 100 patients), and one PPPD pedigree (composing of 90 members including 26 patients), one PM pedigree (cornposing of 34 members including 17 patients). Results While diagnosed as porokeratosis, the five pedigrees included three distinctive variants, each with its own clinic characteristics. The lesions was initiated on the face in DSAP subtype, on palms and the flex-ion side of fingers in PPPD subtype; or involving sun-covered areas in PM subtype. Of the three subtypes of porokeratosis, the onset age in DSAP subtype was earliest, usually about 8-20 years old, about 14-20 years old in PPPD subtype, but PM subtype about 20-30 years old. Conclusions As a group of autosomal dominant genodermatosis, porokeratosis presented various clinic variants with different genetic basis. And, different subtype could be seen in a same patient or same pedigree.
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Objective To evaluate of the clinical efficacy and side-effect of 595nm tunable pulsed dye laser in treatment of children heamangioma of skin. Methods A retrospective study was conducted according to 76 children patients with heamangioma of skin (including 53 cases of nevus flammenus, 20 strawberry hemangioma and 3 cavernous hemangioma) treated with 595nm tunable pulsed dye laser. We evaluated the efficacy and side-effect of the treatment. Results The effectiveness rates were 83.2% and 86.63%, respectively, in nevus flammenus and strawberry hemangioma, but there were disappointed efficacy obtained in patients with cavernous hemangioma treated with 595nm tunable pulsed dye laser. There was 7.9% of the side-effect rate in the children with heamangioma of skin treated by 595 nm tunable pulsed dye laser. Conclusion There is satisfactory clinical efficacy and lower side-effect for children hemangioma treated by 595nm tunable pulsed dye laser.