Objective To observe the expression of free Ca2+ and Angiotensin Ⅱ 1 type receptor(AT1 R)in vascular smooth muscle cells(VSMCs)of patients with hypertensive or normotensive coronary artery diseases(CAD).Methods During the coronary artery bypass graft operation,the surplus saphenous vein of patients who admitted to our Cardiac Surgery Department was collected and cultured.All patients were divided into hypertensive or normotensive group.Free Ca2+ in the cultured human VSMCs was determined by confocal laser scanning microscope(CLSM)after different kinds of AngiotensinⅡ being added,respectively.Total RNA was extracted from cultured VSMCs.Then RT-PCR was conducted for the observation of the expression of AT1R in both groups.Results Ca2+in human VSMCs rapidly increased when stimulated by Angtensin Ⅱ in two groups.After stimulated by Angiotensin Ⅱ,both free Ca2+ level and the expression of AT1R in VSMCs of hypertensive patients were higher than those of the normotensive patients(P＜0.05).Conclusion There are certain changes of free calcium in the cultured human vascular smooth muscle cells,when stimulated by Angiotensin Ⅱ.There are also difierences in AT1R expression between hypertensive CAD patients and normotensive CAD patients.

BACKGROUND:The molecular mechanism of traumatic deep vein thrombosis is complex.Numerous studies focus on clinical observation and epidemiology,but its molecular mechanism has not been a new breakthrough.OBJECTIVE:By use of gene array technology,this study was aimed to study the expression changes of matrix metalloproteinases in rat models of traumatic deep vein thrombosis,and to explore the roles of matrix metalloproteinases in traumatic deep venous thrombosis.METHODS:A total of 150 SD rats,SPF grade,of 8-12 weeks old,body weight of 250-300 g,were divided at random into normal control group (n=10) and model group (n=140).Rat traumatic deep venous thrombosis models were set up by clamping the femoral vein and fixing the bilateral hind limbs,and the fixation of hip spica with plaster bandage was conducted in each group.Then rats were divided into 7 subgroups:post-traumatic 0.5 hours,post-traumatic 2.5 hours (initial period of thrombosis),post-traumatic 25 hours (thrombogenesis at thrombotic crest-time),post-traumatic 25 hours non-thrombogenesis at the thrombotic crest-time),post-traumatic 72 hours (thrombus resolution),post-traumatic 72 hours thrombus insolution) and post-traumatic 168 hours (nonthrombosis).At the corresponding phasess,the femoral vein tissues were incised,and total RNA of femoral vein was extracted using Trizol one-step method.Applying Genechip Rat Genome 430 2.0 genechips,the gene expressions in femoral vein were detected in different groups.The rate of traumatic deep venous thrombogenesis and non-thrombogenesis,the rate of thrombi solution and insolution were observed;the expressions of matrix metalloproteinases and tissue inhibitor of metalloproteinases at different time phases was detected by gene array data analysis.RESULTS AND CONCLUSION:Three model rats died and the remaining 147 rats were involved in the final analysis.At the post-traumatic 25 hours,the rate of thrombogenesis was 50.5% and nonthrombogenesis was 49.5%.To the post-traumatic 168 hours,the rate of thrombus solution was 56.7% and thrombus insolution was 43.3%.Both matrix metalloproteinases and tissue inhibitor of metalloproteinases exhibited differential expressions in the course of traumatic deep venous thrombosis.Under the thrombus insolution state,matrix metalloproteinases continued to show a high expression,tissue inhibitor of metalloproteinase expression was down-regulated in the thrombus formation,was significantly inhibited in the thrombus insoluUon process.In the process of traumatic deep vein thrombosis and insolution,matrix metalloproteinase was closely related to traumatic deep vein thrombosis,the matrix metalloproteinase/tissue inhibitor of metalloproteinases are likely to affect the biological state of thrombosis.

BACKGROUND:There is lack of an effective measuring means to diagnose deep venous thrombosis (DVT) in clinic.KLF2 and KLF4 are down-expressed at prethrombotic state,which may be served as predictive molecular markers to diagnose DVT.OBJECTIVE:To explore the feasibility of KLF2 and KLF4 as molecular markers to prediagnose DVT in rats.METHODS:Totally 90 rats were obtained from 100 rats to establish traumatic DVT models and divided into the prethrombotic,thrombosis crest-time and non-thrombosis groups.The remained 10 rats served as control group.Rat blood was collected at each time point,and the expressions of KLF2 and KLF4 were detected by real-time PCR.RESULTS AND CONCLUSION:The KLF2 and KLF4 mRNA expressions in the prethrombotic group and thrombosis crest-time group were lower than that of the control group.However,the KLF2 and KLF4 mRNA expressions in the non-thrombosis group was higher than that of the control group.Therefore,KLF2 and KLF4 may be candidate molecular markers for prediagnosis of DVT in rats.

BACKGROUND: Deep venous thrombosis (DVT) always occurs after orthopedic surgery. At present, clinical diagnosis of DVT has been lack of an effective measuring means for a long time. Cathepsin may be an effective biological marker of DVT. OBJECTIVE: To study the expression change of cathepsin B and cathepsin C in the rat blood cells before and after DVT and to investigate the feasibility of cathepsin B and cathepsin C as candidate molecular markers for early diagnosis of DVT. METHODS: Totally 100 Sprague Dawley rats were randomly divided into normal control group (n=10) and model group (n=90). Rat traumatic deep vein thrombosis models were established by clamping the femoral vein and fixing the bilateral hind limbs. According to observation time points and the different situations of thrombosis, rat models were assigned to three subgroups: pre-thrombosis, intra-thrombosis, and non-thrombosis. Blood RNA of each group was extracted and reverse transcribed into cDNA. The expression of cathepsin B and cathepsin C in blood cells was detected using real-time fluorescence quantitative PCR. RESULTS AND CONCLUSUON: Expression of cathepsin B and cathepsin C in the blood cells was obviously expressed in the intra-thrombosis subgroup. There was no significant difference in cathepsin B and cathepsin C expression between pre-thrombosis, non-thrombosis groups and normal control group. These findings suggest that cathepsin B and cathepsin C are closely related to DVP and they can be used as the candidate molecular markers for early diagnosis of DVT.

Objective To analyze the clinical information of a family with one patient who have systemic lupus erythematosus (SLE) and Fabry disease,as well as the enzymatic activity and gene mutation in these family members.Methods Clinical characteristics were collected from the proband and her family members.Peripheral blood samples from three members of this family were collected and the enzymatic activity was measured by fluorimetrie substrate assay.Genomic DNA was extracted from one male member with significantly decreased enzyme activity,the 7 exons and their flanking introns of GLA gene were amplified by PCR and directly sequenced.Results The enzyme activity of two family members was significantly decreased,the genetic analysis of the male member revealed a missense mutation in exon 2:c.334C＞T (CGC＞TGC)( p.R112C ).Family members except the proband had no definite evidence to support the presence of SLE.Conclusion The coexistence of SLE and Fabry disease is extremely rare.Immunological test,enzymatic activity and gene mutation analysis seem to be helpful for the differential diagnosis.

OBJECTIVETo discuss arthroscopic technique of single Kirschner wire and suture avoiding epiphyseal line fixation for tibial intercondylar eminence fracture and its clinical results.

METHODSFrom May 2008 to December 2012, 21 patients (13 males, 8 females, ranging in age from 6 to 14 years old) with tibial intercondylar eminence fracture were treated arthroscopically with single Kirschner wire and suture avoiding epiphyseal line fixation technique. According to Meyers and McKeever classification, 7 patients were type II, 10 patients were type III, and 4 patients were type IV. Active rehabilitation began at one week after operation. The patients were followed up for 10 to 30 months. X-ray films were taken to evaluate fracture healing at 1, 3, 6 months after operation; range of motion, the anterior drawer test, the lachman test and the Lysholm knee score were used to evaluate clinical effects.

RESULTSAll fractures were healed without displacement at 6 weeks after operation. Anterior drawer test and the lachman test were both negative in all patients at 3 months after operation. Lysholm knee score was 95.5 ±2.5 at 6 months after operation, and postoperative X-ray film did not find epiphyseal line broadening or narrowing.

CONCLUSIONArthroscopic treatment for tibial eminence intercondylar fracture with single Kirschner wire and 8-shaped suture avoiding epiphyseal line fixation technique has many advantages, such as firm fixation,early mobilization, less invasive, less injury of physis and satisfactory effect.

Adolescent ; Arthroscopy ; Bone Wires ; Child ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Sutures ; Tibial Fractures ; surgery

Objective:To establish an HPLC coupled with post column derivatization method for the determination of gentamicin C components and the related substances based on the latest European Pharmacopeia and compare with the electrochemical method. Methods:A Hydrophilic C18(250 mm ×4.6 mm, 5 μm)column was used with acetonitrile-50 mmol·L-1 sodium hydroxide solution ( pH 2. 6) containing 0. 7% trifluoroacetic acid and 0. 025% pentafluoropropanoic acid (1. 5∶98. 5) as the mobile phase. The temper-ature of post-column reaction was set at 30℃, and the samples were detected by a fluorescence detector withλex of 340nm andλem of 430nm. A pulsed amperometric detector (PAD) was applied in the electrochemical method with golden working electrode in a four-po-tential working mode. Results: According to the results of the two detection methods, the linear range of C1a , C2 , C2a and C1 was 5.82-233.00,6.92-277.00,4.00-160.00and6.23-249.00 μg·ml-1(r >0.9993) , respectively. The limit of detection and quantization were 0. 92-3. 28ng and 1. 37-5. 19ng, respectively. Conclusion:There is no significant difference between the determina-tion results of the two methods.

Objective　To compare the Immunoreactivity of various HEV Antigen with Anti-HEV IgM. Methods Solid-phase enzyme immunoassay( EIA ) was developed for detecting anti-HEV IgM by using synthetic peptides E30, E42, E33, and recombinant antigen from HEV ORF-2. Results Of 60 anti-HEV positive sera by using E30, E42, E33 and recombinant antigen as coating antigen, Anti-HEV IgM positive rates were 76.7%, 26.6%, 18.3% and 66.7% respectively. In Acute-phase and convalescence-phase sera of the patients with Hepatitis E, Anti-HEV IgM positive rate was 90% and 3.3% respectively. Conclusions The HEV E30-based EIA will be very useful in the early diagnosis of Hepatitis E.

Objective To evaluate the relationship between Henoch-Schonlein purpura (HSP) accompanying renal impairment and helicobater pylori(Hp) infection.Methods This study consisted of 304 patients with HSP.The patients were divided into 2 groups(group A and group B) based on Hp infection or not(91 cases in group A and 213 cases in group B).Compared with the rates of accompanying renal impairment in 2 groups.And observed the recovery from renal impairment between the patients who were turned into negative(group C)and patients still were positive after the anti-Hp therapy(group D).Numeration data were analyzed by ?2 test.Results Group A which was with Hp infected,the accompanying renal impairment ratio was 65.9%.Group B which was without Hp infected,the ratio was 35.2%.There was significant difference between 2 groups(?2=24.378 P

Objective To investigate the life quality in infants with food allergy before and after treatment by using of Food Allergy Quality of Life Questionnaire - Parent Form(FAQLQ - PF). Methods The severity of eczema in 46 children with food allergy was determined by eczema area and severity index(EASI). The life quality of such infants was assessed by the FAQLQ - PF after 2 - months treatment. Results (1)Among the 50 subjects,46(92% )respon-ded to the tests;the average age was(4. 78 ± 1. 48)months. Twenty - nine(63. 04% )subjects had family history of allergic disease. Egg(73. 91% )was the most common allergen,followed by cow milk(43. 48% ),fish and shrimp (8. 70% ),and carrot(4. 35% ).(2)The scores of EASI,FAQLQ - PF in children with food allergy before treatment were 13. 20 ± 12. 86 and 2. 12 ± 1. 33,respectively,the scores after treatment declined(2. 02 ± 4. 40 and 1. 33 ± 1. 08, respectively),there was a statistical significance(t = 28. 070,4. 014,all P ﹤ 0. 05).(3)Compared with pre - treat-ment,the scores of emotional impact subscale and food anxiety subscale declined,from 2. 10 ± 1. 47 and 2. 63 ± 1. 83 to 1. 30 ± 1. 09 and 1. 68 ± 1. 37,respectively,and there were statistically significant differences(t = 5. 204,8. 818,all P ﹤ 0. 05).(4)There was a positive correlation between the EASI and FAQLQ - PF scores(ρ = 0. 205,P ﹤ 0. 05). Conclusions Food allergy adversely affects the quality of life in infants. The poor life quality is associated with more severe symptoms. The life quality in infants and parents can be improved by standardized treatment.