Objective To investigate the expression of matrix metalloproteinase 2 (MMP2) and MMP9 in the breast cancer and its relationship with tumor invasive potentiality,node metastasis and prognostic significance.Methods 130 cases of breast cancer and 10 cases of breast benign disease were studied by immunohistochemical staining with MMP2,MMP9 monoclonal antibodies.Results The positive expression rate of MMP2 and MMP9 were 83.08％ (108/130) and 71.54％ (93/130) in the breast cancers,which was significantly higher than those in breast benign disease ( P ＜0.01 ).The expressions of MMP2 and MMP9 obviously increased with tumor sizes and clinical staging( P ＜0.05,P ＜ 0.01 ).Over expression of MMP2 and MMP9 in the breast cancers with axillary lymph node metastases were significantly higher than those without lymph node metastases ( P ＜ 0.01 ).Conclusions The expression of MMP2 and MMP9was closely correlated to lymph node metastasis,and may act as markers for lymph node metastasis and prognostic in breast cancer.

Objective To purify BMSCs by a simple Colony-Forming system and identify BMSCs in vitro.Methods BMSCs were planted at low-density ( 10/cm2 ) in the early stage of isolation,additional cells were scraped until a colony formed,secondly seeded those Colony-Forming cells at low-density.This process was repeat again when passaged cells formed another colony.This so called Colony-Forming selection was repeated several times until highly purified cells were obtained.Biological characters of BMSCs were observed.The surface antigens of BMSCs were identified by flow cytometry.The multipotentiality of BMSCs was assayed for differentiation into either osteoblasts or adipocytes.Results Homogeneous cells were obtained after BMSCs were passaged twice to thrice by Colony-Forming system.These BMSCs highly expressed positive surface antigens of BMSCs ( CD29 in 98.8％,CD90 in 98.4％ ) meanwhile seldom expressed negative surface antigens of BMSCs (CD31 in 2.6％,CD45 in 3％ ).BMSCs efficiently differentiated into osteoblasts and adipocytes.Conclusions Colony-Forming system is a simple and effective way to get highly purified BMSCs in a short time.

Objective To investigate the long-term prognosis and possible risk factors of acute kidney injury(AKI)following cardiac surgeries. Methods Clinical data of 941 patients undergoing open heart surgery from January 2004 to June 2007 were retrospectively analyzed.Survival and renal function of above patients were informed through telephone or clinic follow-up till February 2010.Long-term survival was analyzed using risk-adjusted Cox proportional hazards regression model. Results Of the 941 patients after cardiac surgery,275(29.2%)developed AKI and 666(70.8%)did not.Survival was worse in patients with AKI(67.8%vs 85.6%,P<0.01)and was proportional to its severity (AKIN 1,2 and 3 stages:70.7%,62.3%and 58.6%,P<0.01).Although 90.5% of patients had complete renal recovery after AKI at discharge of hospital,they still had an increased risk for death during follow-up compared with patients without AKI (69.6% vs 85.6%,P<0.01).In the Cox proportional hazards regression model,age(HR=2.238),anemia (HR=1.625),prolonged operation time (HR=1.153),AKI severity (HR=1.473)were independent risk factors for long-term prognosis after cardiac surgery.At the end of the follow-up,patients with AKI had statistically higher Scr than non-AKI patients(107.6 μmol/L vs 83.0 μmol/L,P=0.014),and among those AKI patients,34.0%progressed into CKD 3-5 stages,34.8%developed double serum creatinine. Conclusions AKI is a risk factor for the long-term prognosis of cardiac surgery.Patients with complete renal recovery after AKI still has an increased risk for death and poor renal function compared with patients without AKI.

BACKGROUND:Currently, cellular composition and the features of the nucleus pulposus are stil not to be clarified.
OBJECTIVE:To establish the in vitro culture system of rat nucleus pulposus-derived mesenchymal stem cells and to identify their multi-lineage differentiation potential.
METHODS:Mesenchymal stem cells from the nucleus pulposus tissues of Sprague-Dawley rats were cultured in vitro. Then, cells at passage 3 were induced to differentiate into osteoblasts, adipocytes and chondrocytes as experimental group. cells cultured with basic culture medium served as controls.
RESULTS AND CONCLUSION:cells isolated from rat nucleus pulposus could form the sunflower-like colonies and exhibit clone-like growth when they cultured at a low density. cells at passage 3 became homogeneous and exhibited fibroblast-like morphology. After 28 days of osteogenic induction, arizarin red positive signals were detected in the experimental group. The mRNA expressions of RunX2, osteopontin and osteocalcin were significantly increased in the experimental group, compared to the control group (P<0.05). After 21 days of adipogenic induction, oil red-O positive cells were detected in the experimental group. The mRNA expressions of C/EBPαand PPARγ2 were significantly increased in the experimental group, compared to the control group (P<0.05). After 21 days of chondrogenic induction, safranin O/fast green staining was positive in the experimental group. The mRNA expressions of aggrecan and Col2a1 were significantly increased in the experimental group, compared to the control group (P<0.05). Our findings in this study suggested that nucleus pulposus-derived mesenchymal stem cells could be isolated from the Sprague-Dawley rat nucleus pulposus and exhibited clonal-like growth when they were cultured in vitro. These cells were confirmed to have the potential to differentiate into adipocytes, osteoblasts and chondrocytes in vitro.

ObjectiveThe purpose of this study was to observe the correlation of mTOR and VEGF gene with nephropathy indicators in diabetic rats. Methods Forty-eight male Sprague-Dawley(SD)rats were divided into diabetes mellitus group(DM=28)and control group(NDM=20). Diabetic models were produced by injection of streptozotocin. In the courses of 12,16,20 and 24 weeks,the histology scores(HS)and glomerular basement membrane(GBM)thickness were collected. The protein and mRNA expressions of the gene of mTOR,VEGF and VEGFR2 were observed by immunohistochemistry and real-time quantitative polymerase chain reaction (RT-Q-PCR)by SYBR Green. And the standardized cycle of threshold(SCt)was regarded as the indicators of the mRNA expression. Results HS and GBM thickness were significantly higher in DM rats than those in NDM rats,especially in DM rats of the courses of 20 and 24 weeks(P < 0.01). IHC scores of VEGF and VEGFR2 were higher in total DM rats and were positively correlated with each other. There were positive correlations between HS with VEGF and VEGFR2 in total DM rats(P < 0.05). SCts of VEGF and VEGFR2 were significantly higher and were positively correlated with each other in total DM rats(P < 0.01). SCt of VEGF and GBM thickness showed positive correlation in total DM rats. SCt of VEGF was highest in the course of 12w DM rats. SCt of VEGFR2 gradually decreased following by the diabetic course,and was lowest in the course of 24w. There were no significantly differences in IHC scores and SCt of mTOR between DM and NDM rats. But the IHC scores of mTOR,VEGF and VEGFR2 were positively correlated with each other and with HS in total DM rats(P < 0.05). Conclusion HS and GBM thickness were higher in diabetic rats,especially in the course of 24w,which could reflect the injury of nephropathy. The protein and mRNA of VEGF and VEGFR2 were high expressed in kidney of DM rats and increased with the increasing of diabetic course. The mRNA expression of VEGF was positively correlated with GBM thickness of in diabetic nephropathy(DN). The protein expressions of mTOR,VEGF and VEGFR2 might have synergistic effects in DN of DM rats. But the results could not exclude the influences of different courses,sample size and experimental conditions.

Objective To investigate the value of urinary liver-type fatty acid-binding protein (L-FABP),neutrophil gelatinase-associated lipocalin (NGAL) and their combination in predicting the development and the severity of acute kidney injury (AKI) following cardiac surgery in adults. Methods Scr,urinary L-FABP and NGAL corrected by urine creatinine at preoperation,0 h and 2 h postoperative time points were examined.The differences of above indexes between AKI and non-AKI groups were compared.Receiver operating characteristic (ROC)curves and area under curves (AUC) were used to evaluate the diagnostic value of urinary L-FABP,NGAL and their combination for AKI. Results The cohort consisted of 109 patients,26(23.9％) developed AKI,and AKIN stage Ⅰ, Ⅱ and Ⅲ was 46.2％,34.6％ and 19.2％ respectively.Levels of urinary L-FABP and NGAL were significantly higher in AKI patients at 0 h and 2 h postoperatively.AUC to predict AKI or AKI stage Ⅱ-Ⅲ was 0.81 to 0.87 using either of the biomarkers.The performance of combining two biomarkers was better with AUC of 0.911 to 0.927. Conclusions Urinary L-FABP and NGAL increase at the early stage after cardiac surgery.Combination of these two biomarkers enhances the accuracy of the early diagnosis of postoperative AKI after cardiac surgery before a rise of Scr.

Objective To observe the effects of electroacupuncture combined with edaravone on the conduction velocity of sciatic nerve and oxidative stress in rats with diabetic peripheral neuropathy. Methods 60 Sprague Dawley (SD) rats were included. 10 of them were selected as normal group. The other rats were modeled as diabetic peripheral neuropathy with streptozotocin. 48 of them were randomly selected and divided into electroacupuncture group (n=12), edaravone group (n=12), electroacupuncture + edaravone group (n=12), and model group (n=12). The threshold temperature for wave tail was tested, the levels of superoxidase dismutase (SOD) and malonaldehyde (MDA) were determined, and the conduction velocity of sciatic nerve were measured, before, and 4 and 8 weeks after modeling. Results 8 weeks after modeling, the conduction velocity and SOD increased in the electroacupuncture group, edaravone group and electroacupuncture+edaravone group compared with the model group (P<0.01), with the MDA decrease (P<0.01), while it was improved more in the electroacupuncture+edaravone group than in the electroacupuncture group or the edaravone group (P<0.01). Conclusion Both electroacupuncture and edaravone can inhibit oxidative stress and improve nerve conduction velocity of the sciatic nerve in rats with diabetic peripheral neuropathy, and it is more effective of combination.

Objective To validate the effect of Renji acute kidney injury score (RAKIS) on predicting patients with acute kidney injury (AKI) after cardiac surgeries,and make comparison with Cleveland score,simplified renal index (SRI) and acute kidney injury following cardiac surgery (AKICS).Methods Patients undergoing open heart surgery from 2008/01/01 to 2010/10/31 in Renji hospital were enrolled,and their scores of those four scoring models were calculated.AKI patients were diagnosed by KDIGO,and those scores of AKI patients and non-AKI patients were compared.Receiver operating characteristic (ROC) curve and area under curve (AUC) were used to decide the predictive values of those models.Results A total of 1126 patients were chosen in this cohort,with the average age of (58.43±14.88) years (rang from 18 to 88).The male to female ratio was 1.47:1.And 355(31.5％) patients were developed AKI.AKI stage Ⅰ,Ⅱ and Ⅲ were 65.4％,23.7％ and 11.0％ respectively.RAKIS was significantly higher in AKI patients than in non-AKI patients (17.5 vs 9.0,P ＜ 0.001).The AUCs of RAKIS to predict AKI,AKI Ⅱ-Ⅲ stages,renal replacement therapy (RRT)and in-hospital death were 0.818,0.819,0.800 and 0.784 respectively.The AUCs of Cleveland score and SRI were 0.659 to 0.710,lower than those of RAKIS and AKICS.AKICS had lower value for predicting AKI and AKI Ⅱ-Ⅲ stages (AUC 0.766 and 0.793),but good value in predicting RRT and inhospital death after surgery (AUC 0.804 and 0.835) as compared with RAKIS.Conclusions RAKIS is valid and accurate in the discrimination of KDIGO defined AKI patients,while for predicting the composite end point,AKICS may be more useful.

Objective To explore the clinical value of serum ferritin (SF),erythrocyte sedimentation rate (ESR) and erythrocyte average indexes [mean corpuscular volume (MCV),mean corpuscular hemoglobin (MCH),mean corpuscular hemoglobin concentration (MCHC)] in the prognosis evaluation of patients with small cell lung cancer (SCLC).Methods A total of 72 SCLC patients were enrolled in the Haian People's Hospital Affiliated to Nantong University from January 2013 to October 2016 (SCLC group),and 80 health controls were selected at the same time (control group).The levels of serum SF,ESR and erythrocyte average indexes in SCLC group and control group were detected,and their relationships with clinical features,prognosis and survival time were analyzed.Results The serum levels of SF,ESR,MCV,MCH and MCHC in SCLC patients were (309 ±59) μg/L,(16 ±4) mm/h,(104 ± 12) fl,(32 ±4) pg and (307 ±21) g/L,respectively.The serum levels of SF,ESR,MCV,MCH and MCHC in control group were (186 ±26) μg/L,(15 ±5)mm/h,(85 ± 7) fl,(30 ± 3) pg and (335 ± 25) g/L,respectively.Compared with the control group,the patients in SCLC group were significantly increased on the levels of SF (t =14.168,P ＜ 0.001) and MCV (t =6.143,P ＜ 0.001),and were significantly decreased on the level of MCHC (t =-4.220,P =0.003).There were no significant difference in the levels of ESR (t =1.931,P =0.102) and MCH (t =1.220,P =0.313) between the two groups.The serum levels of SF and MCV were significantly correlated with the stage of SCLC (t =-4.092,P =0.009;t =-4.985,P ＜ 0.001).Multivariate logistic regression analysis showed that high serum SF (OR =5.31,95％ CI:3.09-9.31,P ＜ 0.001) and MCV (OR =1.78,95％ CI:1.10-3.08,P =0.013) were independent risk factors of SCLC.Survival analysis showed that the survival time of the patients in the high SF group was significantly shorter than that in the low SF group (6 months vs.20 months;x2 =6.556,P =0.001).Conclusion Serum ESR,MCH and MCHC levels are not significantly correlated with SCLC,but serum SF and MCV levels are of important clinical significance in evaluating the prognosis of SCLC patients.

Objective:To investigate the clinical efficacy of posterolateral approach combined with anteromedial approach in the treatment of trimalleolus fracture.Methods:A retrospective analysis was performed of the 20 patients who had been admitted to The Second Department of Orthopedics, The First People's Hospital of Tianshui for trimalleolus fractures from January 2016 to August 2020. They were 16 men and 4 women, aged from 20 to 70 years (average, 49.6 years). The lateral malleolus, posterior malleolus and medial malleolus were treated with reduction and internal fixation using the posterolateral approach combined with the anteromedial approach. Postoperative complications were observed, and the foot function was assessed using the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and pain visual analog scale (VAS).Results:In this cohort, the operation time ranged from 85 to 115 minutes, averaging 88.4 minutes and the intraoperative blood loss from 50 to 600 mL, averaging 120 mL. All patients were followed up for 12 to 20 months (mean, 14.5 months). The fracture healing time ranged from 3.2 to 5.4 months, averaging 3.8 months. Follow-ups observed no such complications as infection or necrosis of surgical incision, failure of internal fixation, nonunion, or malunion. The AOFAS ankle-hindfoot score at 12 months after operation (87.8±6.4) was significantly higher than that before operation (32.3±4.9) ( t=29.454, P<0.001); as for VAS, one case scored 0, 13 cases 1 to 3 points and 6 cases 4 points. Conclusion:In the treatment of trimalleolus fracture, a combination of posterolateral approach and anteromedial approach can lead to definitely positive efficacy because of a significant reduction in operation time, intraoperative bleeding and postoperative complications.