Objective To explore the relationship between the level of matrix metallo-proteinase(MMP)-2 ,MMP-3 ,MMP-9 and matrix metallo-proteinase inhibitor-1(TIMP-1) in the synovial fluid of the patients with knee joint osteoarthritis (OA) and the de-gree of articular cartilage injury and prognosis .Methods 52 patients(knee OA group) were given arthroscopic debridement com-bined with sodium hyaluronate ,diacerein .The levels of MMP-2 ,MMP-3 ,MMP-9 and TIMP-1 in synovial fluid were detected in 52 patients with knee OA and 10 normal controls(control group) by enzyme-linked immunosorbent assay .The degree of cartilage inju-ry was assessed with arthroscopy .Results The level of MMP-2 ,MMP-3 ,MMP-9 and TIMP-1 in knee OA group were significantly higher than those of the control group(all P<0 .01) .The levels of MMP-2 ,MMP-3 ,MMP-9 and TIMP-1 in the synovial fluid were positively correlated with the degree of articular cartilage injury ,and the levels of MMP-2 ,MMP-3 ,MMP-9 and TIMP-1 decreased with the improvement of the disease .Conclusion The measurement of MMP-2 ,MMP-3 ,MMP-9 and TIMP-1 have a certain signifi-cance to early diagnosis ,degree judgment and prognostic evaluation of knee OA .

Objective The purpose of our study was to report the results of using arthroscopic Remplissage and Bankart repair in patients who had an engaging Hill-Sachs lesion with significant glenoid bone loss. Methods We retrospectively reviewed 49 consecutive patients who underwent arthroscopic Remplissage and Bankart repair for anterior shoulder instability with a mean duration of follow-up of 29.0 months (24-35 months). At the time of surgery the mean age of 42 men and 7 women was 28.4 years. All patients were diagnosed as recurrent anterior shoulder dislocation with a bony lesion of glenoid and an engaging HillSachs lesion. An arthroscopic Remplissage and Bankart repair using metal anchor was performed in all cases.ASES score, Constant-Murley score and Rowe score were used to evaluate the stability and the function of the shoulder. Results Patients' active forward elevation significantly(P=0.007) improved from 162.9°±17.1°preoperatively to 170.9°±7.4° at final follow-up. The external rotation was 56.0°±17.6° before the surgery compared with the 54.1°±17.1° postoperatively(P=0.511 ). The ASES score, Constant-Murley score and Rowe score was 84.7±11.3, 93.3±8.7 and 36.8±8.5 preoperatively compared with 96.0±3.4, 97.8±3.6 and 89.8±12.5 postoperatively. Significant difference could be found with regard to ASES score (P=0.000), ConstantMurley score (P=0.005) and Rowe score (P=0.000). One redislocation happened and a subluxation was noticed in three patients (8.3%). Conclusion Arthroscopic Remplissage combined with Bankart repair can achieve satisfactory for recurrent anterior shoulder dislocation accompany with engaging Hill-Sachs lesion.

Objective To evaluate the clinical results of arthroscopic repair of massive rotator cuff tear. Methods The study involved 16 patients with massive rotator cuff tears treated arthroscopically from September 2007 to June 2009. There were 6 males and 11 females at average age 61.5 years (45-75 years). The rotator cuff tears was repaired with arthroscopic double-row reconstruction. The range of motion, pain, strength of flexed elevation and function evaluation score were all recorded before operation and at final follow-up. The results were evaluated by t test and compared according to age and course of disease. Results All patients were healed without complications and the outcome was improved significantly ( P < 0.01 ). The mean VAS score was improved from preoperative 5.6 to postoperative 1.7,the average forward flexion from 69. 1°to 151.2°, the average external rotation from 14.7° to 32.2°, and internal rotation from L1 level to T10, the mean Constant-Murle from 39 to 85, the mean UCLA from 10.4 to 28, the mean SST from 2.8 to 8.8 and the strength of flexed elevation from 10.7% of normal side to 65.0%. Compared with preoperation, there was statistical difference in aspects of pain, range of motion, muscle strength and function in postoperation (P < 0.01 ). Conclusion Arthroscopic doublerow fixation can attain satisfactory results in repair of massive rotator cuff tear.

Objective To investigate the relationship of gene polymorphisms of angiotensin eonvertion enzyme (ACE), aldosterone synthase (CYP11B2)and α-adducin with subclinical renal lesion. Methods I/D polymorphism of ACE gene, -344T/C polymorphism of CYP11B2 gene and 460G/T polymorphism of α-adduein gene were detected by polymerase chain reaction (PCR) and restrictive fragment length polymorphism(RFLP) in 604 normotensive subjects and 1081 primary hypertensive patients whose creatinine (Cr) were less than 2mg/L. The primary hypertensive and normotensive subjects were divided respectively into normal group (Ccr≥60ml/min) and subclinical renal lesion (Ccr＜60 ml/min) group, according to creatinine clearance rate (Ccr) calculated by Cockcroft-Gault equation. Results ANOVA, contingency X2 and partition of chi-square were selected. The frequencies of different genotypes of ACE, CYP11B2, and α-adducin were in agreement with Hardy-Weinberg equilibrium in our study. Normal renal function group (A group, n=512) and subclinical renal lesion group (B group, n=92) in normotensive subjects, and normal renal function group (C group, n=828) and subclinical renal lesion group (D group, n=252) in hypertensive patients were compared. The patients in B and D groups were older than those in A and C groups (P<0.01). But there were no significant differences in the age between B and D groups, and between A and C groups. The frequency of ACE-DD genotype in D group was the highest (22.6%) among four groups and the frequency of α-adducin-TT genotype in A group was the lowest (13.3%) among four groups (all P<0.01). The differences of genotype frequencies of ACE and α-adducin genes among other three groups were not significant. No significant difference was found in frequencies of genotypes of CYP11B2 among four groups. Conclusions Subclinical renal lesion is increased with the aging. ACE-DD genotype is related with hypertension and subclinical renal lesion, while α-adducin-TT genotype is related with hypertension and subclinical renal lesion. Association between the genotypes of CYP11B2 and subclinical renal lesion is not found.

Objective To investigate the effects of the popliteofibular ligament (PFL) and/or the popliteus tendon (POP) reconstruction in an external rotation injury model of the knee.Methods Six nonpaired cadaveric knees were tested under the following POP and PFL states:intact,sectioned,and reconstructed using 3 different techniques.Each knee was subjected to 5 N·m external rotation torque at flexion angles of 0°,30°,45°,60°,90°,and 120°.A navigation system was used to measure the motion changes of the tibia with respect to the femur.Results The external rotation increased 2.1°±0.7° (2.0°-2.3°) after sectioning only PFL while flexing the knee from 30° to 120°,and increased 1.3°±1.2° (0.5°-2.0°) after sectioning only POP while flexing the knee from 30° to 120°.There was no significant difference between two groups.There was significant increase in external rotation as 4.1°±1.6° (2.8°-5.0°) after sectioning both POP and PFL.Comparing the POP and POP+PFL reconstruction techniques to the intact state,there were significant differences.The external tibial rotation in POP or POP+PFL reconstruction group was significant different from that in intact or PFL reconstruction group while flexing the knee from 30° to 90°.Conclusion In a LCL-intact PLC injury model,the POP and PFL function as a unit in resisting external rotation.All surgical procedures we described and tested are able to reduce the increased external rotational laxity found in the sectioned state.The PFL reconstruction is able to restore external rotation to near normal.However the techniques involving POP reconstruction over constrained external rotation during laxity testing.

Objective To compare stability between the traditional endoscopic technique and thetibial inlay in treatment of posterior cruciate ligament (PCL) reconstruction.Methods Between April 2005 and December 2009,135 patients underwent surgery for PCL in multiple-ligament injuries,and only 88 (65.2％) patients were followed up longer than 2 years.The follow-up time ranged 24 to 77 months (mean,45.9 months).Fifty-seven cases (64.8％) were treated with endoscopic transtibial PCL reconstructions,and 31(35.2％) received tibial inlay.Each patient was evaluated using the Tegner,Lysholm,and American Academy of Orthopaedic Surgeons (AAOS) knee-rating scales,KT-1000 arthrometry and Telos.Results The preoperative KT-1000 measurement was (13.5±4.8) mm (transtibial) and (13.7±5.2) mm (inlay).The postoperative KT-1000 measurement was (2.4±3.4) mm (transtibial) and (2.2±3.6) mm (inlay).The preoperative Telos measurement was (14.9±7.1) mm (transtibial) and (14.9±5.9) mm (inlay).The postoperative Telos measurement was (4.6±4.0) mm (transtibial) and (4.3±3.9) mm (inlay).There were significant differences (P＜0.01) in KT-1000 and Telos between pre-and post-operative measurement,while it does not reach statistical significance (P=0.880,0.956,0.744,0.647) in KT-1000 and Telos when comparing transtibial and inlay technique.There were no significant differences in sex,age,time from injury to surgery,combined injuries or Lysholm,Tegner,AAOS knee score between two groups.Conclusion There was no important advantage of one technique over the other.Transtibial or inlay technique for PCL reconstruction can restore anteroposterior knee stability.

Objective To evaluate the effectiveness of anterior cruciate ligament (ACL) reconstruction combined with anterolateral ligament(ALL)reconstruction in ACL injury patients with high-grade pivot shift.Methods From May 2015 to April 2016,156 patients underwent ACL reconstruction by the same surgeon,and 22 of them with grade 2/3 pivot shift were included in this study.Anteroposterior knee stability was evaluated using KT1000 measurement,and the rotatory stability was assessed using the pivot-shift test.The Lysholm score was used to monitor the clinical function.Results There were 14 male subjects and 8 females,with an average age of 29.3 years.The mean follow-up period was 8.6 months.The mean side-to-side difference of anteroposterior knee laxity was 2.1 ± 0.6 mm,significantly improved compared with the preoperative 8.9 ± 3.1 mm.The preoperative pivot-shift indicated 2+ in 20 patients and 3+ in 2 patients,while at the final follow-up,21 patients had negative pivot shift with 1 of 1+ pivot shift.The difference was significant.The average Lysholm score improved significantly from 60.5 ± 12.3 preoperatively to 79.2 ± 7.8 at the final follow-up.Conclusion The rotatory instability can be effectively restored through the ACL reconstruction combined with ALL reconstruction in patients with high-grade pivot shift.The early-stage knee stability and functional outcomes indicate significant improvement postoperatively.

Objective To study the effect of enhanced MK gene expression in hepatic carcinoma cells.Methods The recombinant plasmid pIRES2-EGFP-MK was transfected into SMMC 7721 cells.The mRNA and protein expression levels of MK gene in these cells were determined by real-time PCR,Western blotting and flow cytometry.The intracellular DNR accumulation of these cells was measured by flow cytometry.To investigate the effect of MK gene mediated multidrug resistance,MTT assay was employed to determine the cellular sensitivity of different chemotherapeutic drugs in MK-overexpressed SMMC 7721 cells.Results The mRNA and protein expression levels of MK gene significantly increased after the recombinant plasmid pIRES2-EGFP-MK transfected into SMMC 7721 cells,suggesting that the recombinant plasmid pIRES2-EGFP-MK can enhance the transcription of MK effectively.The DNR accumulation of MK transfected cells decreased significantly (4.06 ± 0.88,P ＜ 0.05),and IC50 of MK transfected cells to ADM/5-FU increased significantly (15 ± 3,27 ± 4,P ＜ 0.05).Conclusions After the recombinant plasmid pIRES2-EGFP-MK transfected into hepatic carcinoma cells,expression of midkine increased,enhancing the resistance of hepatic carcinoma cells to chemotherapeutic drugs.

【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-Tj^a 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-Tj^a 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-Tj^a-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.

【Objective】 To explore the application of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) in the genotyping of difficult blood typing samples, and to provide evidence for clinical blood transfusion. 【Methods】 Three ambiguous blood group samples, submitted to Shanghai Blood Center by Shanghai regional hospitals, were studied, of which Sample1 included the proband and his parents. Serological methods were used to perform blood group typing, direct antibody test, unexpected antibody screening and identification test. Blood group genotyping was performed by using the MALDI-TOF MS detection systeme stablished in our laboratory. Sanger sequencing was used to confirm gene mutation sites, and serological or flow methods were used to verify specific samples′ phenotype. 【Results】 Serological results indicated the existence of antibodies against high frequency antigens in sample 1 (including proband and her mother), 2 and 3. The genotyping results of MALDI-TOF MS showed that the proband of sample 1 was Di(a+ b+ ), her father was Di(a-b+ ), her mother was Di(a+ b-), sample 2 was p, and sample 3 was Jr(a-). Sequencing results of three samples were consistent with mass spectrometry typing results. Serological results showed that sample 2 had a p phenotype. The flow cytometry results suggested that sample 3 had a Jr(a-) phenotype. 【Conclusion】 For the first time, we applied MALDI-TOF MS technology to blood type genotyping of ambiguous clinical samples in China. Compared with other genotyping methods such as PCR-SSP, MALDI-TOF MS has the advantages of rapid detection, high throughput and high specificity, which would contribute to identification of difficult blood typing samples in the future, as well as rare blood group screening.