1.Preventive Application of Antimicrobial in Orthopedic Patients During Perioperative Period: A Survey and Analysis
Chinese Journal of Nosocomiology 2004;0(10):-
OBJECTIVE To find out the situation of the use of preventive application of antimicrobial in orthopedic perioperative patients and to evaluate the rationality of use. METHODS To make a survey of the use of antimicrobials in orthopedic perioperative patients during Apr-Jun 2004 and to evaluate the kinds,frequency,combined administration,course,effectiveness,etc. RESULTS 99.2% patients had received antimicrobials involving 14 kinds.62.8% from them used one.34.5% used two and 2.7% used three. CONCLUSIONS It is necessary to strengthen the management of preventive antimicrobial in orthopedic patients during perioperative period.
2.Application of orthogonal design in the development of Echinococcosis ELISA kit and its evaluation
Yanhong LI ; Yuehong GONG ; Zhaoxia ZHANG
Chinese Journal of Laboratory Medicine 2014;37(7):544-547
Objective To select the optimum conditions of Echinococcosis ELISA kit and kit preparation by using orthogonal experiments,then to test their detection performance.Methods Taking the absorbance of the samples as the indicator,orthogonal experiments of three factors four levels were taken to optimize the conditions of hydatid disease detection ELISA kit.Through L16 (43)orthogonal experiments,the concentration of the antigen(A),the dilution multiple of the sample (B) and the dilution multiple of the enzyme labeled antigen (C) were studied.The specimen were provided by The First Affiliated Hospital of Xinjiang Medical University specimen library collected between March 2011 and June 2013,including 36 sera confirmed alveolar Echinococcus by the gold standard,56 sera confirmed Echinococcus granulosus disease by the gold standard and 72 sera as the control group(including healthy people,cirrhosis,hepatitis,etc).Detecting sensitivity and specificity were compared using F test for statistical analysis.Results Orthogonal design showed the size proportion of three factors of echinococcosis ELISA kit:antibody dilution > sample dilution > antigen coating concentration.The optimal preparation conditions were A1 B2 C4,that was,the concentration of antigen was 1 mg/L,sample dilution of 1∶ 100,dilution of secondary antibody of 1∶160 000.Through the 3-factor and 4-level orthogonal design using F-test analysis,the fact of the concentration of the kit antigen(A),F =1.181,P =0.393; the fact of the dilution multiple of the sample (B),F =2.544,P =0.152,the fact of enzyme labeled antigen dilution (C) F =2.544,P =0.039.The sensitivity of the kit to Echinococcus granulosus disease and alveolar Echinococcus were 83% (30/36),and 91% (51/56),respectively; the specificity was 91% (29/32) and 83% (33/40); the misdiagnosis rate of 17% (6/36) and 9% (5/56) ; the misdiagnosis rate 9% (3/32) and 18% (7/40) ; positive likelihood ratio 8.86% (83/9) and 5.20% (91/18) ; negative likelihood ratio of 0.18% (17/91) and 0.11% (9/83).Conclusion Orthogonal design is a good method that can find out optimal conditions of preparation for Echinococcosis ELISA kit and improve the test performance.
3.An experimental study on effects of tranilast on prophylaxis of a chronic constriction mononeuropathy
Yuehong BAI ; Guiqiu SHI ; Longhai ZHANG
Chinese Journal of Orthopaedic Trauma 2002;0(04):-
Objective To clarify the actions of tranilast,an anti allergic drug,on chronic constriction mononeuropathy through an experiment using an animal model according to the method of Bennett et al. Methods 36 rats were divided into 2 groups: one was treated with tranilast(200mg/kg?d-1,p.o every experimental day),the other was controlled with solvent only. Chronological changes of heat evoked withdrawal latencies on a hot plate,nerve coducting velocitics, and histopathological changes were compared between the two groups from one to four weeks after loose ligation in the sciatic nerve. Results The changes of the withdrawal latency and nerve conducting velocities in the tranilast treatment group were significantly longer than those in the control. The rats without tranilast treatment showed inflammation in and around the constricted nerve bundles,axonal degeneration,phagocytes invasion and interstitial edematous changes at 7 days,numerous axonal sprouts and remyelination at 14 days,and regeneration in the nerve bundles at 28 days. In contrast,the rats treated with tranilast showed less inflammation or nerve fiber degeneration,and better regeneration than those of the control. Conclusion The actions of tranilast appear to have beneficial effects on prophylaxis of a chronic constriction mononeuropathy and on preservation of the nerve functions in rats.
4.CLONING AND EXPRESSION OF SOLUBLE HCV NS5B GENE IN EUKARYOCYTES
Yuehong ZHANG ; Huijuan DUAN ; Liancai JU
Medical Journal of Chinese People's Liberation Army 2001;0(08):-
HCV NS5B, acting as a RNA dependent replication enzyme,has emerged as an attractive protein used as a target for screenig of drugs against HCV NS5B, and plays an important role in HCV replication. In the report the gene expression of NS5B in E.coli was investigated. PCR was performed to gain the gene of HCV NS5B from plasmid pBRTM/HCV 1 which contains whole sequence of HCV, and the truncated NS5B gene containing no hydrophobic domain was cloned into pGEM Teasy vector. The gene of the truncated NS5B was cut from pGEM Teasy vector and cloned into E.coli expression plasmid pET 21b, then pET 21b NS5B was transfected into E.coli cells. The protein E.coli lysates were purified and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting assay. The RdRp activity of NS5B was examined by scintillation proximity assay (SPA). The truncated NS5B gene was successfully cloned into pET 21b. The results of SDS PAGE and Western blotting assay showed: ①the molecular weight of the expressed product was about 68 000 D, ②The truncated NS5B protein was existed in media of E.coli cells, ③The activity of NS5BDCT21 His from HCV 1b amounted to 6 900 cpm (total incorporation of approximately). These findings suggest that soluble NS5B can be successfully expressed in E.coli and could secret into media.
5.Applied research of selective bronchial arterial embolization for severe hemoptysis
Yuehong GONG ; Gang CHANG ; Weifang PEI ; Xiaoyan ZHANG ; Hongfu LI
Chinese Journal of Postgraduates of Medicine 2012;35(23):16-18
Objective To observe the clinical curative effect of selective bronchial arterial embolization for severe hemoptysis.Methods Thirty-eight hospitalized patients with severe hemoptysis was enrolled in this study.Of which 31 patients were recurrent hemoptysis,27 patients were with bronchiectasis,4 patients were with bronchial lung cysts,4 patients had unknown causes,3 patients were with tuberculosis.All patients were diagnosed by chest imaging examination,fiber bronchoscopy,bronchial arteriography,and they were all treated by selective bronchial arterial embolization.Results The most times of embolization was 3,and bronchial arterial imaging were performed for vascular malformation.After having bronchial arterial embolization,35 patients were immediately released of bleeding.Postoperative 24 h,8 patients had hemoptysis again,of which 1 case was performed with conservative treatment and subsequently had pneumonectomy,5 patients had effective conservative treatment,2 cases were performed embolization again after failed conservative treatments.Hemoptysis completely disappeared within 1 week.The total effective rate was 97.4%(37/38).Patients were followed up for 1 year,of which 13 patients relapsed again,11 patients'annual and each time's quantity of hemoptysis reduced 50%.Two patients had pneumonectomy after a number of embolization.The total effective rate was 94.7% (36/38).After treatment,3 patients had mild chest pain in short time,2 patients had shoulder pain,3 patients had chest tightness,and 3 patients had subcutaneous passive congestion.After being treated for short-term,these symptoms disappeared eventually.Conclusion Selective bronchial arterial embolization for the treatment of acute or recurrent severe hemoptysis is very effective,and can avoid the risk of surgery.It is effective for recurrent cases and worthy of clinical application.
6.Vasculature anatomy in medial foot and its clinical application in foot and ankle reconstruction
Jianse ZHANG ; Shuhong SUN ; Yuehong ZHUANG ; Songqing LIN
Chinese Journal of Trauma 2012;(10):933-936
ObjectiveTo investigate anatomical characteristics of the medial foot vessels and effects of different vascular pedicle skin flaps in repair of foot and ankle trauma.MethodsThirty adult cadaveric lower limbs were injected with red latex through the popliteal artery and posterior tibial artery to anatomically observe the cutaneous arterial origin,branches,distribution and anastomosis of the medial foot.Then,anterior medial malleolar artery perforator flaps and distally-based medial foot flaps were harvested and used for repairing foot and ankle trauma of 16 patients.Results The origin of cutaneous blood vessels of the medial foot was diversified and mainly included the anterior medial malleolar artery,medial tarsal artery,and arterial arcades anastomosing with anterior posterior branches of the two former arteries and the superficial branches of plantar digital artery and the medial plantar artery.According to distribution area of the anterior medial malleolar artery and the medial tarsal artery,the vascular anatomy of the medial foot skin was classified into three types.Clinically,all the flaps survived.Follow-up ranged from 2 weeks to 20 months,which showed normal color,good shape and good pain and warm sensation of the flaps.ConclusionThe anterior medial malleolar artery perforator flaps present good reconstruction of soft tissue defects around the ankle,whereas the distally-based medial foot flaps present good reconstruction of soft tissue defects of the mid-forefoot.
7.Analysis on severe complications of percutaneous radiofrequency ablation for hepatic malignancy
Neng WANG ; Guojun QIAN ; Qiang SHEN ; Yuehong SHENG ; Lei ZHANG
Chinese Journal of Hepatobiliary Surgery 2011;17(3):186-188
Objective To study the prevention and treatment of severe complications of percutaneous radiofrequency ablation(PRFA)for hepatic malignancy. Methods A series of 939 patients with primary hepatic carcinoma or hepatic metastasis confirmed by pathological examinations or clinical manifestations underwent 1098 treatments of PRFA between January 2006 and December 2009. All the patients were followed up to study the short-or long-term complications related to PRFA. Results Complications developed in 9 patients: bile duct injury (4 patients), hemothorax (2 patients), and intra-abdominal hemorrhage (3 patients). The incidence of complication was 0.82% (9/1098) and the complication-related mortality was 11.1% (1/9). Conclusions Although PRFA which is minimally invasive, is a safe and effective treatment, there were still risks for this procedure, especially when the tumor is located at the portahepatic region or the patient has coagulopathy. Some serious complications can be prevented. It is important to observe the strict indications for RFA and to carry out the procedure carefully. Early detection of complication is important.
8.Effect of 99Tcm-labeled mouse double minute 2 antisense oligonucleotide on target gene expression of prostatic cancer cells
Qiong WU ; Yuehong ZHANG ; Peng FU ; Guomei TIAN ; Changjiu ZHAO
Chinese Journal of Nuclear Medicine and Molecular Imaging 2014;34(2):125-129
Objective To investigate the effect of mouse double minute 2 (MDM2) mRNA ASON and mismatched oligonucleotide (ASONM) radiolabeled with 99Tcm on target gene expression in LNCaP cells.Methods The ASON and ASONM targeted to MDM2 mRNA were synthesized and radiolabeled by 99Tcm with the bifunctional chelator of HYNIC.The labeling efficiency,radiochemical purity,stability and molecular hybridization activity were investigated.The different concentrations of 99Tcm-HYNIC-ASON (0,100,500 nmol/L) and 99Tcm-HYNIC-ASONM (500 nmol/L) coated with lipofectamin 2000 were incubated with prostate cancer cells for 24 h,then RT-PCR and Western blot were carried out to assay the MDM2,p53 mRNA and the corresponding protein level.The variables of RT-PCR and Western blot were analyzed using one-way analysis of variance and q test.Results The labeling efficiency of ASON and ASONM were (65.15± 2.05)% (n=5) and (64.93±2.18)% (n=5),respectively.The radiochemical purity were both more than 90%.99Tcm-HYNIC-ASON had a good stability and could hybridize to the sense oligonucleotide (SON).The contents of MDM2 mRNA in 0,100,500 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups were 0.458±0.035,0.250±0.026,0.174±0.032,0.463±0.033,respectively,and there were significant differences between each 2 groups except between 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups (F=33.69,q =24.32-91.45,all P<0.01).The average density of MDM2 protein in the 4 groups were 90.712±3.042,71.218±2.915,32.775±3.062,88.121±2.710,respectively (F=235.93,q=6.43-19.14,all P<0.01; except 0 nmol/L99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).The contents of p53 mRNA in the 4 groups were 0.185±0.046,0.203±0.040,0.213±0.027,0.163±0.049,respectively(F =2.18,P> 0.05).The average density of p53 protein was 33.865 ± 2.213,70.445±2.180,99.025±3.012,38.351±3.271,respectively (F=53.98,q =3.32-6.74,all P<0.01 ; except 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).Conclusions The MDM2 antisense probe can accumulate in the prostate cancer cells,and specially hybridize to the MDM2 mRNA and inhibit target gene expression.This novel molecular probe has a promising potential for the diagnosis of prostate cancer at gene level.
9.Effects of rat interleukin-10 gene treatment on expression of collagen, MMP13 and TIMP1 in fibrotic rats
Yuehong HUANG ; Yunxin CHEN ; Lijuan ZHANG ; Zhixin CHEN ; Xiaozhong WANG
Chinese Journal of Immunology 2014;(5):613-617
Objective:To study the effects of rat interleukin-10 (rIL-10) gene treatment on the expression of collagen , matrix metalloproteinase 13(MMP13) and their specific inhibitors the tissue inhibitor of metalloproteinase 1(TIMP1) in porcine serum in-duced liver fibrosis rats then to explore the anti-fibrotic effect of rL-10.Methods:Thirty SD rats were divided into normal control and fibrosis model group.Normal control group (group C) was intraperitoneally injected with 0.5 ml normal sodium twice a week for 8 week, while the fibrosis model group was injected with equal volume of pig serum for 8 week.At the beginning of the 5th week, fibrosis model group was further randomly divided into a fibrosis model subgroup ( group M ) , rIL-10 gene treatment subgroup ( group I ) and empty vector control subgroup(group P).Rats in group C and M were injected with Ringer’s solution as a reagent control via the tail vein weekly, rats in group I were injected with the rIL-10 plasmid pcDNA3-rIL-10, and rats in group P were injected with empty vector pcDNA3.All rats were sacrificed at the end of 8th week, and the liver tissue samples were collected to observe deposition of collegan in liver tissue by sirius red staining and detected the expression of MMP 13 and TIMP1 in the liver tissue by SP immunohistochemistry .Re-sults:Sirius red staining showed that the area of the collegan deposition was dramatically increased in fibrosis model subgroup and emp -ty vector control subgroup compared with the normal control group , and the area of the collagen deposition was dramatically decreased in rIL-10 gene treatment subgroup compared with the fibrosis model and empty vector control subgroup .Immunohistochemistry analysis showed that the expression of MMP 13 and TIMP1 in fibrosis model subgroup and empty vector control subgroup was significantly higher than the normal control group , but compared with normal control group , expression of MMP13 was significantly increased and expres-sion of TIMP1 was significantly decreased in rIL-10 gene treatment subgroup .Compared with fibrosis model subgroup and empty vector control subgroup, the expression of MMP13 and TIMP1 was dramatically decreased in rIL-10 gene treatment subgroup.Conclusion:rIL-10 gene treatment attenuates the area of collagen deposition in liver fibrosis rats associated with downregulation of TIMP 1.
10.Antisense imaging targeting mouse double minute 2 oncogene in prostate cancer xenografts
Yuehong ZHANG ; Changjiu ZHAO ; Qiong WU ; Peng FU ; Guomei TIAN
Chinese Journal of Nuclear Medicine and Molecular Imaging 2014;34(1):48-52
Objeetive To explore the value of antisense imaging of 99Tcm-labeled ASON targeting mouse double minute 2(MDM2) mRNA for the diagnosis of human prostate cancer.Methods The ASON targeting MDM2 mRNA and the mismatched oligonucleotide (ASONM) were synthesized and radiolabeled with 99Tcm using the bifunctional chelator HYNIC.The labeling efficiency and radiochemical purity were investigated.Animal models of nude mice bearing human prostate cancer LNCaP were established and divided into 3 groups with 10 mice in each group.99Tcm-HYNIC-ASON,99Tcm-HYNIC-ASONM (study groups) and 99TcmO4-(control group) were injected at the dose of 7.4 MBq through the tail vein,respectively.Tumor imaging was acquired with SPECT and the tumor-to-muscle (T/M) ratio was measured.The data was compared by one-way analysis of variance.Results The labeling efficiencies of ASON and ASONM were (65.15± 2.05) % and (64.93±2.18) %,respectively.Their radiochemical purity was greater than 90%.At 1,4 and 10 h post injection,the T/M ratios of 99Tcm-HYNIC-ASON group were 3.217±0.125,3.749± 0.201 and 4.028±0.186,and those of 99Tcm-HYNIC-ASONM group were 1.579t0.128,1.715±1.140 and 1.683±0.139,and control group 2.146±0.132,1.847±0.124,1.528±0.152,respectively.The T/M ratios in control group and 99Tcm-HYNIC-ASONM group were significantly lower than those in 99Tcm-HYNICASON group at 1,4 and 10 h,respectively (F=213.37-235.41,t=3.527-4.738; all P<0.01).The T/M ratios of 99Tcm-HYNIC-ASONM group and control group were not significantly different at 1,4 and 10 h (t=2.154,2.287 and 2.236,all P>0.05).Conclusion The antisense probe of MDM2 can accumulate specifically in prostate cancer tissue in animal models,which might be useful as a non-invasive genetic tool for the early diagnosis of prostate cancer.