1.Establishment of FMEA risk management procedures for blood coagulation tests in clinical laboratories
Guanghua NIU ; Yujie GAO ; Baihui CUI
Chinese Journal of Laboratory Medicine 2016;(1):13-17
Definiting the workflow and key link of the risk management in medical laboratory by FMEA.Identifying risk factors of the workflow and key link of blood coagulation test by the criteria for laboratory accreditation , such as ISO15189 recognition criteria and CAP laboratory accreditation inspection . Through the evaluation of the blood coagulation test , effective corrective actions and examining performance data periodically , the quality of the blood coagulation test can be improved continuously.
2.Total hip arthroplasty with structural bone-grafting for high congenital dislocation of adult hip
Shuguang GAO ; Guanghua LEI ; Kanghua LI
Orthopedic Journal of China 2006;0(23):-
[Objective] To discuss the effect of total hip arthroplasty with structural bone-grafting for high congenital dislocation of adult hip.[Method]From August 2003 to October 2006,22 patients(29 hip)with congenital dislocation of hip were treated with total hip arthroplasty and structural bone-grafting.There were 10 males and 12 females.There were 13 left and 26 right.The ages were from 23 to 42 years old,with the average of 34.6 years.Clinical symptoms included hip pain,hip instability and limping.The preoperative average Harris scores was 46.8.The preoperative average length difference between two legs was 3.8 cm.The femoral head dislocation height from normal position was 3.9 cm on average(range,3.2 to 5.6cm).The average range of motion of the hip:flexion 66.5?,abduction 23.8?,external rotation 20.4?,internal rotation 5.3?.Posterolateral approach was used during operation and all the acetabular cups were reconstructed at the true acetabular location and were covered completely with femoral head autograft.The reduction of prosthesis was achieved by releasing surrouding soft tissue.The acetabular reconstruction was done with the cemented acetabular component in 18 hips,the small acetabular component in 5 and common cementless acetabular component in 6.Cementless shaft was used in 10 hips,common cemented shaft in 13,steno-vertical cemented shaft in 4 and long cemented shaft in 2.The clinical effects were evaluated with operative time,blood loss,Harris scale,joint range of motion,length difference between two legs and complications.[Result]The average operative time was 95 minutes(range from 70 to 135 rain).The average blood loss was 400 ml(range from 300 ml to 650 ml).Primary healing of wound was observed in all patients without infection.Position of prothesis judged by X-ray was fine at 1 week postoperatively,the average abduction angle of the cup was 39?,and the average anterior angle was 13?.Twenty-two cases were followed up from 10 months to 47 months(average 17.6 months)after surgery,the average score was 89.2 points(ranged from 72 points to 93 points),according to the evaluate of Harris.The average range of motion of the hip was 115.3? for flexion,44.6? for abduction,49.5? for external rotation,26.8? for internal rotation.The preoperative average length difference between two legs was 1.2 cm.There were 2 postoperative femoral nerve palsy which resolved completely within 4 months.Femoral head autografts were not absorbed.Neither loosening nor dislocation of the prostheses occurred in the time of follow-up.[Conclusion]Total hip arthroplasty with structural bone-grafting is an effective method for high congenital dislocation of adult hip.It improves symptoms,functions and shapes.Structural bone-grafting can provide reliable acetabular coverage and restore bone stock.In case of the altitude of femoral head dislocation upward from normal position less than 4cm,reduction of prosthesis can achieve by releasing surrouding soft tissue without femoral nerve palsy.
3.Application of optical coherence tomography in interventional diagnosis and treatment of borderline coronary artery disease
Zhigang HAN ; Guanghua GAO ; Rui WANG
Chinese Journal of Interventional Cardiology 1993;0(02):-
Objective To evaluate the feasibility and efficacy of optical coherence tomography (OCT) in interventional diagnosis and treatment of borderline coronary artery disease. Methods Sixteen 40%-70% angiographically stenotic lesions from 15 patients were enrolled in the current study. Intravascular OCT was applied to each lesion after informed consent was obtained to evaluate stenosis percentage, size of lipid core, thickness of fibrous cap, existence of plaque rupture, and presence of thrombosis or calcification. Therapeutic strategies were made according to OCT outcomes as well as clinical symptoms and electrocardiographic changes. Stent coverage, apposition, and tissue prolapse between stent struts were determined in the patients undertaken percutaneous coronary intervention(PCI). Results Qualitative OCT images were obtained in 14 lesions. Ten lesions were greater than 50% stenosis, with large lipid core and fibrous cap thinner than 65 ?m, and plaque ruptures were noted in 2 of the lesions. PCI were performed in these 10 lesions. Two lesions underwent predilatation and significant intima tearings and dissections were detected with OCT. Repeated OCT after stent implantation showed complete coverage in all 10 lesions. Focal incomplete apposition were noted in 2 lesions. Significant tissue prolapse occurred in 3 lesions and segmental incomplete deployment of stent in 2 lesions. The remaining 4 lesions had small lipid cores, thick fibrous caps but with no plaque ruptures, thus PCI was not performed. Conclusion Intracoronary OCT is an effective tool to determine features of coronary lesions. It plays an important role in diagnosis of vulnerable lesions, strategy-making in treating borderline coronary artery disease and evaluation of immediate result of PCI.
4.Gene cloning and expression purification of human autoimmune antigen SSA/60
Guanghua NIU ; Cheng ZHANG ; Dan LYU ; Yujie GAO
International Journal of Laboratory Medicine 2016;37(16):2224-2225,2228
Objective To clone human autoimmune antigen SSA/Ro60 and to purify its expression to provide the material basis for the assisted diagnosis of human autoimmune diseases .Methods The SSA/Ro60 gene was cloned by RT‐PCR and directionally inserted into expression vector pPICZ .The recombinant plasmid was transformed into Pichia SMD1168 .The obtained recombinant protein was identified by SDS‐PAGE and Western blotting .Results The amplified full‐length sequence was about 1 .5 kb in size . The pPICZ‐SSA positive clone produced a 60 × 103 recombinant protein which had natural immunogenicity of human autoimmune antigen SSA/Ro60 by SDS‐PAGE and Western blot .Conclusion Human autoimmune antigen SSA/Ro60 is successfully cloned and expressed ,which lays a foundation for diagnosing autoimmune diseases .
5.Relationship between DNA of Rat Kidney and Post by Comet Assay
Junfeng JIN ; Cuilian GAO ; Yuchuan CHENG ; Guanghua LUO
Journal of Sun Yat-sen University(Medical Sciences) 2009;30(4):386-389
[Objective] To study the correlation between kidney cell DNA degradation and postmortem interval within the span of 6-48 hour after the subject rats′ death.[Methods] To select 18 healthy mature female SD rats and equally divide them into 6 groups.To execute the rats with cervical spine articulation and put the rats under the incubator temperature of 25.1℃ (the average temperature of the 5 previous Decembers in Guangzhou prefecture).Sample kidney tissue from the rat separately 0 hour,6 hours,12 hours,24 hours,36 hours,48 hours,and 60 hours after the rats′ execution to prepare monoplast suspension,which is committed to comet assay.The comet images were captured by fluorescence CCD.Kinetic Comet 4.0 software was used to analyze images.Relevant data were collected by kinetic Comet 4.0 software and were subjected to Kruskal-Wallis test.[Results] Within the postmortem interval of 6-48 h,the number of SD rat kidney cell DNA fragments increased as the postmortem interval lengthens.So did the comet tail length.The Oliver tail moment and tail DNA of comet also showed sign of increase in positive proportion to the postmortem interval (their values corresponding to 60-hour-postmortem-interval were not obtainable.Kruskal-Wallis test indicated:the discrepancies of TL among the 6 groups were all significant (P < 0.01).The difference of TM between 6 h group and 12 h group was not significant (P > 0.05).The difference of TM between 24 h and 36 h was significant (P < 0.05).The difference of TDNA among 6 h,12 h,and 36 h groups were not significant (P > 0.05).The difference of TDNA between 36 h and 48 h was significant (P < 0.05).[Conclusion] Degradation of nuclear DNA of the rat kidney cells increases as the postmortem interval lengthens and comet assay may provide important empirical evidence for determining the postmortem interval.
6.A postmortem semi-quantitative study on time-dependent changes of DNA content in chondrocytes in rat's costal cartilage
Haowei XING ; Yuchuan CHEN ; Jianding CHENG ; Cuilian GAO ; Guanghua LUO
Chinese Journal of Forensic Medicine 2009;24(6):365-367
Objective To study the relationship between the DNA content of chondrocytes in the costal cartilage and postmortem interval in putrefactive rat cadavers.Methods Nuclear DNA wag visualized by modified Feulgen's staining method.DNA content of ehondrocytes in the costal cartilage was semi-quantita tively determined by a computerized image analysis system in rats within 35d postmortem.Results Staining intensity of the nuclei was gradually reduced within from 1d to 28d postmortem.The nuclej could not be detected at 35d.The DNA content of chondrocytes decreased time-dependently within 28 days after death as determined semi-quantitatively,which revealed a linear relationship between DNA content and postmortem interval.Conclusion DNA content of chondrocytes in the costal cartilage reduces time-dependently with the extension of postmortem interval.
7.Comparison of effects of different anesthesia on immunologic function during perioperative radical mastectomy in patients having undergone preoperative chemotherapy
Guanghua ZHANG ; Jincheng LI ; Li CHEN ; Lubo GAO ; Bing HUO
Chinese Journal of Anesthesiology 2009;29(7):588-591
Objective To compare the effects of different anesthesia on immunologic function during perioperative radical mastectomy in patients having undergone preoperative chemotherapy. Methods Sixty ASA Ⅰ or Ⅱ patients, aged 28-64 yr, weighing 55-70 kg, scheduled for radical mastectomy, were studied. Thirty patients received preoperative chemical therapy for 3-4 weeks were randomly divided into CP group and CS group ( n = 15 each); thirty patients received no chemical therapy were also randomly divided into NCP group and NCS group (n=15 each).Propofol at a rate of 4-6 mg·kg-1·h-1 was infused in group CP and CS and sevoflurane with the end-tidal concentration of 1.5%-2.5% inhaled in group NPC and NCS to maintain anesthesia. Peripheral venous blood samples were taken before chemical therapy, before anesthesia, immediately after operation and at 72 h after operation for determination of the levels of T lymphocyte subsets and NK cells and plasma CK19 mRNA expression (by RT-PCR). CD4+/CD8+ ratio and the micrometastatic rate of the tumor cells were calculated. Results The CD3+ , CD8+ and NK cell level were significantly lower before anesthesia, immediately after operation and at 72 h after operation than before chemical therapy ( P < 0.05). The CD8+ immediately after operation and NK cell level at 72 h after operation in group CP, and CD4+ and NK cell levels immediately and at 72 h after operation in group CS were significantly lower than those before anesthesia ( P < 0.05). CD4+ and NK cell levels immediately and at 72 h after operation were significantly lower in group CS than in group CP, and in group NCS than in group NCP (P < 0.05). The CD4+/CD8+ ratio was significantly lower immediately after operation in group CS than in group CP ( P < 0.05) .There was no significant difference in the micrometastatic rate of the tumor cells among all groups (P > 0.05). Conclusion Sevoflurane combined anesthesia has a stronger inhibitory effect on immunologic function in radical mastectomy patients with preoperative chemotherapy than propofol combined anesthesia.
8.Unconditional logistic regression analysis of risk factors of type 2 diabetes mellitus in Dalian
Xiaohong GAO ; Guirang SONG ; Ping XIN ; Li MA ; Hong WEN ; Ping ZHANG ; Zhengnan GAO ; Guanghua SONG
Chinese Journal of Tissue Engineering Research 2005;9(11):190-192
BACKGROUND: Diabetes mellitus is a chronic metabolic disease caused by various factors,such as environmental factor,inherited factor,etc.,and its cause isn't very clear now. This study aims to investigate the risk factors mentioned above in the onset of diabetes mellitus, and is of significance in the first and second grade of prevention of diabetes mellitus.OBJECTIVE: To investigate the risk factors of type 2 diabetes mellitus to provide evidence for the proper intervention of it.DESIGN: Cross-sectional study based on diagnosis.SETTING: Department of epidemiology in a university and department of endocrinology in a university hospital.PARTICIPANTS: The subjects were residents who have resided in town and country of Dalian for more than 5 years and were elder than 40 years old. The method of stratified cluster random sampling was carried out among natural persons who were divided into two groups according to city and country. Totally 2 500 persons were taken in total and 1 250 persons were from each group. The subjects who had been diagnosed as diabetes mellitus were included.METHODS: Questionnaire survey was employed for all the subjects, including physical examinations such as height,weight,waist girth,hip girth,blood pressure,blood glucose,etc. Patients of type 2 diabetes mellitus were taken as the case group and the normal was as the control group. The unconditional univariate and multivariate logistic regression were used.MAIN OUTCOME MEASURES: Unconditional logistic regression analysis of risk factors of type 2 diabetes mellitus with single factor analysis and multivariate analysis.RESULTS: Family history of diabetes mellitus(OR = 2.339),obesity[body mass index(BMI),OR = 1.462],systolic pressure(OR = 1.016),hyperlipidemia(OR = 1.615), age(OR = 1.043) were the major risk factors for type 2 diabetes mellitus.CONCLUSION: Family history of diabetes mellitus, increase of systolic pressure,obesity,high blood lipid and age are the risk factor for type 2 diabetes mellitus. For all communities,control of body mass and high blood pressure is an important way to prevent type 2 diabetes mellitus.
9.Bcl-2 and Bax expression in mice with collagen-induced arthritis following heterogenic umbilical cord blood stem cell transplantation
Guanghua NIU ; Yujie GAO ; Xu SUN ; Jing DU ; Cairong MING ; Mingli GAO
Chinese Journal of Tissue Engineering Research 2012;16(10):1856-1860
BACKGROUND: The occurrenceand development of rheumatoid arthritis were strongly associated with unbalance proliferation and apoptosis of synovial cells and lymphocytes. Some synovial cell apoptosis was abnormal. OBJECTIVE: To observe effects of heterogenic double umbilical cord blood stem cell transplantation on Bcl-2 and Bax expression in mice with type Ⅱcollagen-induced arthritis. METHODS: C57BL/6(H-2b) mice were induced with Freund's complete adjuvant and type Ⅱcollagen to establish mouse models of type Ⅱcollagen-induced arthritis.At 2days after secondary immunity incubation, mice were injected with saline via tail vein in the model and normal control groups. Umbilical cord blood hemopoietic stem cells were injected into mouse tail vein in the UBSCs transplantation groups (single dose:2×106/50 g; double dose: 1×106/50 g each, totally 2×106/50 g). Mice were intragastrically administrated methotrexate in the methotrexate positive control group, 0.017 5 g/kg once, once every 5 days, totally six times. RESULTS AND CONCLUSION: Joint tissue below knee and elbow was obtained at42 days following transplantation. Histopathology displayed that smooth and glossy articular surface, no inflammatory cell infiltration in the synovial layer and normal chondrocytes in the normal control group. Hyperplasia, a lot of inflammatory cell infiltration and damaged cartilage surface were visible in the model group. Slight hyperplasia and a little inflammatory cell infiltration were detectable in the methotrexate positive controlgroup and single UBSCs transplantation group. Double UBSCs transplantation group exhibited smooth and glossy articular cartilage surface, no damage, a little inflammatory cell infiltration. Immunohistochemistry demonstrated that Bcl-2 and Bax expression was lower in the double UBSCs transplantation group compared with single UBSCs transplantation group (P < 0.05). Compared with normal control group, no significant difference was detected (P > 0.05). Results suggest that double umbilical cord blood stem cells can induce synovial cell apoptosis in mice with type Ⅱcollagen-induced arthritis in a certain number and action time, and protect synovial membrane against damage.
10.Matrix metalloproteinase-2 and matrix metalloproteinase-9 mRNA expression in mice with collagen-induced arthritis following heterogenous umbilical cord blood stem cell transplantation
Guanghua NIU ; Yujie GAO ; Jing DU ; He GUO ; Baishan WANG ; Mingli GAO
Chinese Journal of Tissue Engineering Research 2012;16(19):3529-3534
BACKGROUND: Matrix metalloproteinase (MMP) degrades extracellular matrix, which is a necessity of joint destruction in rheumatoid arthritis patients. MMP-2 and MMP-9 can evaluate rheumatoid arthritis and serve as an index to predict progressive destruction of the joint. OBJECTIVE: To observe heterogenous allogeneic umbilical cord blood stem cell (UBSC) transplantation on MMP-2 and MMP-9 expression in the spleen of mice with type Ⅱ collagen-induced arthritis. METHODS: Fetus cord blood was sterilely obtained and cord blood stem cells were separated. The C57BL/6(H-2b) mice were assigned to five groups (n=10). Except normal control group, models of collagen-induced arthritis were established using complete Freund's adjuvant + type Ⅱ collagen. Mice from the methopterin group were intragastrically administered methopterin suspension 0.017 5 g/kg, once every 5 days. Other groups used caudal vein injection. Mice from the model and normal control groups were injected with saline. Mice from the mono-UBSCs group and double-UBSCs group were injected with 2×106/kg UBSCs from one and two parents. At 42 days following injection, animals were sacrificed and the ankle joint was obtained for histopathological detection. MMP-2 and MMP-9 mRNA expression in the spleen was examined using reverse transcription-polymerase chain reaction. RESULTS AND CONCLUSION: Double-UBSC transplantation could significantly inhibit inflammatory cell infiltration in synovial tissue of mice with type Ⅱ collagen-induced arthritis, repaired impaired cartilage tissue. The repair effect was better than that in methopterin group and mono-UBSCs group. MMP-2 and MMP-9 mRNA expression in the spleen was significantly lower in the double-UBSCs group than the mono-UBSCs group (P < 0.01). These suggest that heterogenous allogeneic double-UBSCs transplantation participated in pathological changes in rheumatoid arthritis cartilage and in synthesis of cartilage extracellular matrix and effectively treated rheumatoid arthritis by regulating MMP-2 and MMP-9 mRNA expression.