ObjectiveTo approach the regulatory mechanism of high mobility group box-1 (HMGB1) on the expression of E-selectin in human umbilical vein endothelial cell (HUVEC).Methods Homeobox A9 (HOXA9) siRNA was transfected to HUVEC at logarithmic phase, real-time fluorescence quantitative polymerase chain reaction (real-time qPCR) and Western Blot were used to determine the HOXA9 mRNA expression and protein expressions; a blank control group and a nonsilence negative control group were set. HUVEC stable transfected with pRNA-u6.1/Neo-HMGB1 shRNA plasmids (HUVEC with low-expression HMGB1) was obtained, and HOXA9 and E-selectin mRNA expressions were determined with real-time qPCR; a nonsilence transfection group served as the negative control. The HOXA9 siRNA was transfected to HUVEC with low-expression HMGB1 as co-transfection group, and the E-selectin expressions was determined with real-time qPCR; a HMGB1 shRNA group and a HOXA9 nonsilence group served as control.Results① HOXA9 mRNA (2-ΔΔCT) and protein expression (integralA value) in blank control group were 1.094±0.115 and 1.031±0.060. Compared with nonsilence transfection group, HOXA9 siRNA transfection group could significantly reduced mRNA and protein expression of HOXA9 [HOXA9 mRNA (2-ΔΔCT): 0.257±0.030 vs. 1.035±0.091,t = 14.010,P = 0.002; HOXA9 protein (integralA value): 0.278±0.042 vs. 0.975±0.014,t = 27.310, P = 0.002].② Compared with nonsilence transfection group, HMGB1 shRNA transfection could up-regulate HOXA9 mRNA expression in HUVEC (2-ΔΔCT: 2.519±0.278 vs. 0.856±0.063,t = 10.100,P = 0.001), also could down-regulate E-selectin mRNA expression (0.311±0.046 vs. 1.080±0.201,t = 7.415,P = 0.000).③ Compared with HOXA9 nonsilence group and HMGB1 shRNA group, HMGB1 shRNA and HOXA9 siRNA co-transfected HUVEC cells could significantly elevate E-selectin mRNA expression (2-ΔΔCT: 3.445±0.428 vs. 1.085±0.212, 1.004±0.104,t1 = 8.507, t2 = 9.603, bothP< 0.001).Conclusion HMGB1 may regulate E-selectin expression through the HOXA9 regulation in HUVEC.

Objective To observe the change of collagen in osteoarthritis and study the pathogenesis of osteoarthritis.Methods The right hind limb of twenty-four adult New Zealand White rabbits were immobilized with plaster cast in extension position for 10,20,30 and 40 days,respectively.Six animals without immobilization served as control.The articular cartilage of the medial femur condyle was harvested for transmission electron microscopy,in-situ hybridization of collagenⅡ,and immunohistochemistry of collagenⅠ,Ⅱ,Ⅲ.Results Transmission electron microscopy showed articular cartilage was destructed from the fine collagen fiber network of tangential zone.The fine collagen fiber network did not contain chondrocyte.Immunohistochemistry and in-situ hybridization showed that in earlier period of osteoarthritis,the collagen typeⅡand its gene expression firstly increased,then decreased with destruction of ultrastructure,and chondrocytes enhanced type Ⅱ collagen expressing and synthesizing mainly in transition zone and upper deep zone.In articular cartilage of osteoarthritis there was type Ⅲ collagen,instead of typeⅠcollagen.Conclusion In osteoarthritis,articular cartilage degenerated from tangential layer,in which collagen can not be repaired after destruction,this may contribute to the chondral degeneration.

An enantioselective method was developed for the separation and determination of three chiral hexabromocyclododecanes ( HBCDs ) including α-HBCD, β-HBCD, γ-HBCD in soil and earthworm by HPLC-ID-MS/MS. d18-HBCDs used as internal standards were added to the samples before extraction. HBCDs enantiomers were extracted from soil by accelerated solvent extraction ( ASE ) with n-hexane/DCM (1:1,V/V) at 100℃ and 10 MPa for 5 min, and further cleaned up using silica column. HBCDs enantiomers were extracted from earthworm by vortex turbulence with ethyl acetate. The extracts were orderly sulphonated by sulfuric acid, and purified by silica column. For all HBCDs enantiomers, good linearities were obtained in the concentration range of 0. 25-50 ng/mL. Limits of detection ( LOD) and limits of quantification ( LOQ) were 0. 00544-0. 00766 ng/g and 0. 0173-0. 0244 ng/g, respectively in soil. The recoveries of spiked samples at 0. 05 and 2. 5 ng/g levels were 80. 0%-95. 9% with relative standard deviations ( RSD, %) of 5. 7%-11. 9% in soil. Limits of detection (LOD) and limits of quantification (LOQ) were 0. 0103-0. 0148 ng/g and 0. 0328-0. 0471 ng/g, respectively in earthworm. The recoveries of spiked samples at 0. 1 and 5 ng/g levels were 78. 0% -94. 4% with relative standard deviations ( RSD, %) of 6. 1% -12. 2% in earthworm. This method can meet the requirements of determination of trace HBCDs in soil and earthworm.

BACKGROUND:Such methods as transplanting autologous periosteum or autologous bone marrow mesenchymal stem cells (BMSCs) can promote the repair of articular cartilage defects for sure. But they all have their own limits in chondrogenic abilities,which results in an unsatisfactory curative effect.OBJECTIVE:To study the effect of transplanting BMSCs (which were induced into chondrocytes) combined with autogenous periosteum on repairing articular cartilage defects in rabbits.MATERIALS:A total of 18 New Zealand rabbits,aged 6-8 months,were divided with random digits table method into 3 groups,namely,periosteum+BMSCs group,periosteum group and blank control group,with 6 ones (12 knee joint samples) in each group. METHODS:In periosteum+BMSCs group,BMSCs were harvested and adherently cultured with trypsin digestion method. Then they were induced by transforming growth factor 81 into chondrocytes. At the same time,immunofluorescence labeling was performed to BMSCs membranes with PKH-26. Full-thickness articular cartilage defects (diameter:3mm,depth:3mm) were made to bilateral condylus medialis femoris of all rabbits. In periosteum+BMSCs group and periosteum group,defects were covered by homolateral autogenous proximal tibia periosteums,with germinal layer facing to cavitas medullaris. After that,the periosteum+BMSCs group received 3 sutures,followed by injection of 20 μL BMSCs suspension (1×109/L) into the defects,after which the last suture was taken. The periosteum group underwent coverage with periosteum on defects only. The blank control group underwent perforate only.MAIN OUTCOME MEASURES:General observation,histological observation,Wakitani's score,immunohistochemical and in situ hybridization detection of collagen type Ⅱ were performed to defects at 6 and 12 weeks following operation.RESULTS:No sutured periosteums were found desquamate. In periosteum+BMSCs group,defects were filled with hyaline cartilage-like repairing tissues at week 6 following operation;Week 12 following operation saw remodeled tissues whose cells were mainly the implanted cells labeled with PKH-26. In periosteum group,repairing tissues in defect areas were ivory white,smooth with light introcession and distinctively different from the surrounding normal cartilage tissue. In the blank control group,clearer introcession or irregular appearance,even broken surrounding cartilage tissues could be seen in the defect area. Both Wakitani's score and histological score were highest in periosteum+BMSCs group at week 6 and 12 following operation (P＜0.05),with higher ones in periosteum group than in the control group (P＜0.05). What'more,matrix around cells in the repairing tissues showed positive results to both immunohistochemical and in situ hybridization staining of collagen typeⅡ,which proved that cells in repairing tissues were the implanted ones.CONCLUSION:Transplanted BMSCs (which were induced into chondrocytes) combined with autogenous periosteum can form hyaline cartilage-like repairing tissues through which articular cartilage defects are repaired.

Knee osteoarthritis (KOA) is the most common knee joint disease. Although KOA belongs to a disease of degeneration of knee joint cartilage, its pathology involves cartilage, subchondral bone, meniscus, synovial membrane, articular capsule and other joint tissue. The infrapatellar fat pad (IPFP), an intracapsular but extrasynovial structure, has some special characteristics of endocrine metabolism, then it has double roles in the development of KOA, but its protective effect is much more than harmful effect. Considering protective roles of IPFP in KOA and some serious complications after IPFP resection, the surgeon shall protect IPFP as far as possible if total knee arthroplasty surgical field is good. If it is necessary to improve the surgical field, its fibrotic tissue even all part can be removed.
Adipose Tissue ; physiology ; Arthroplasty, Replacement, Knee ; Humans ; Knee Joint ; physiopathology ; surgery ; Osteoarthritis, Knee ; surgery

Objective: To observe the influences of different follow-up methods on rehabilitation and compliance of patients with severe scar after burns. Methods: From January 2012 to May 2016, medical records of 116 patients with severe scar after burns who were admitted to our unit, discharged after wound healing and conforming to the criteria, were retrospectively analyzed. They were divided into face-to-face follow-up group [n=59, 45 males and 14 females, aged (36±9) years] and routine follow-up group [n=57, 44 males and 13 females, aged (35±9) years] based on different follow-up methods they received. On the day of discharge and in post discharge month (PDM) 1, 3, and 6, the Vancouver Scar Scale (VSS) was used to evaluate the hypertrophic scar in joints, Activities of Daily Living (ADL) scale was used to evaluate the disability of patients in the 2 groups. In PDM 1, 3, and 6, Medical Compliance Behavior Questionnaire was used to investigate the medical compliance behaviors of patients in the 2 groups. Data were processed with chi-square test, t test with Bonferroni correction, and analysis of variance for repeated measurement. Results: (1) The VSS score of patients in face-to-face follow-up group on the day of discharge was (11.1±0.7) points, which was close to (11.7±0.7) points of routine follow-up group (t=2.021, P>0.05). The VSS scores of patients in face-to-face follow-up group in PDM 1, 3, and 6 were (10.5±0.6), (8.6±0.7), and (4.7±0.5) points, which were significantly lower than (11.4±0.7), (10.9±1.0), and (9.4±0.8) points of routine follow-up group respectively (t=2.034, 2.033, 2.042, P<0.05 or P<0.01). (2) The ADL score of patients in face-to-face follow-up group on the day of discharge was close to that of routine follow-up group (t=1.781, P>0.05). The ADL scores of patients in face-to-face follow-up group in PDM 1, 3, and 6 were higher than those of routine follow-up group respectively (t=9.683, 8.584, 9.772, P<0.01). (3) The compliance rates of consisted rehabilitation, reasonable diet, and timing consultation of patients in face-to-face follow-up group were better than those of routine follow-up group respectively (χ²=19.015, 13.251, 8.652, P<0.01). Conclusions Compared with routine follow-up by phone, face-to-face follow-up can do better in evaluating the scar condition and ADL of patients with severe scar after burns, and improve the medical compliance rates of patients, which is worthy of clinical promotion.

Objective: To explore the effects of multidisciplinary comprehensive intervention in integrated treatment of patients with extremely severe burns. Methods: One hundred and ten patients hospitalized in our center from July 2013 to August 2017 met the criteria for inclusion in this study, and their medical records were retrospectively analyzed. According to the medical model at that time, 56 patients hospitalized from July 2013 to July 2015 received routine comprehensive treatment led by doctors, and they were included in the conventional intervention group. From August 2015 to August 2017, 54 patients were treated with integrated multidisciplinary interventions performed by a team consisting of physicians, intensive care nurses, burn nurses, intravenous infusion nurses, wound stoma nurses, and rehabilitation therapists, and they were included in the integrated intervention group. Acute Physiological and Chronic Health Assessment Ⅱ (APACHE Ⅱ) scores on admission and 4 weeks after treatment, incidences of pressure injury, wound sepsis, lung infection, and catheter-related infection during hospitalization, length of stay in intensive care unit (ICU) and total length of hospital stay, and Abbreviated Burn Specific Health Scale (BSHS-A) scores at discharge and 3 months after discharge were analyzed. Data were processed with t-test and chi-square test. Results: The APACHE Ⅱ scores of patients in the two groups were close on admission (t=0.573, P>0.05). Four weeks after treatment, the APACHE Ⅱ scores of patients in the two groups were obviously lower than those on admission within the same group (t=5.697, 4.853, P<0.01), and the score of the integrated intervention group was obviously lower than that of the conventional intervention group (t=2.170, P<0.05). No pressure injury was observed in patients of any group during hospitalization. The incidences of wound sepsis, lung infection, and catheter-related infection of patients in the integrated intervention group were 18.5% (10/54), 3.7% (2/54), and 9.3% (5/54), respectively, significantly lower than 42.9% (24/56), 21.4% (12/56), and 26.8% (15/56) of the conventional intervention group, χ²=4.073, 6.075, 3.962, P<0.05. The length of stay in ICU of patients in the integrated intervention group was (50±5) d, obviously shorter than (62±4) d of the conventional intervention group (t=2.852, P<0.01). The total length of hospital stay of patients in the integrated intervention group was (115±8) d, obviously shorter than (140±7) d of the conventional intervention group (t=16.885, P<0.01). At discharge and 3 months after discharge, the scores of BSHS-A, physical function, psychological function, and general health status of 50 patients in the integrated intervention group were significantly higher than those of 48 patients in the conventional intervention group (t=2.886, 3.126, 2.416, 2.544, 2.033, 3.471, 2.588, 2.210, P<0.05 or P<0.01), while the scores of social function of patients in the two groups were close (t=1.570, 1.350, P>0.05). Conclusions Integrative treatment of patients with extremely severe burns by multidisciplinary comprehensive intervention can significantly alleviate the severity of burns, reduce the incidences of wound sepsis, lung infection, and catheter-related infection, shorten the length of stay in ICU and total length of hospital stay, and improve the quality of life.

PURPOSE: The purpose of this study was to investigate the changes of tumor infiltrating lymphocytes (TILs) between core needle biopsy (CNB) and surgery removed sample (SRS) in early stage breast cancer patients and to identify the correlating factors and prognostic significance of TILs changes. MATERIALS AND METHODS: A retrospective study was carried out on 255 patients who received CNB and underwent surgical resection for invasive breast cancer. Stromal TILs levels of CNB and SRS were evaluated respectively. Tumors with ≥50% stromal TILs were defined as lymphocyte-predominant breast cancer (LPBC). Clinicopathological variables were analyzed to determine whether there were factors associated with TILs changes. Log-rank tests and Cox proportional hazards models were used to analyze the influences of TILs and TILs changes on survival. RESULTS: SRS-TILs (median, 10.0%) were significant higher than CNB-TILs (median, 5.0%; p<0.001). Younger age (<60 years, p=0.016) and long surgery time interval (STI, ≥4 days; p=0.003) were independent factors correlating with higher TILs changes. CNB-LPBC patients showed better breast cancer-free interval (BCFI, p=0.021) than CNB-non-LPBC (CNB-nLPBC) patients. Patients were categorized into four groups according to the LPBC change pattern from CNB to SRS: LPBC→LPBC, LPBC→nLPBC, nLPBC→LPBC, and nLPBC→nLPBC, with estimated 5-year BCFI 100%, 100%, 69.7%, and 86.0% (p=0.016). nLPBC→LPBC pattern was an independent prognostic factor of worse BCFI (hazard ratio, 2.19; 95% confidence interval, 1.06 to 4.53; p=0.035) compared with other patterns. CONCLUSION: TILs were significantly higher in SRS than in CNB. Higher TILs changes were associated with younger age and long STI. Changing from nLPBC to LPBC after CNB indicated a worse BCFI, which needs further validation.
Biopsy, Large-Core Needle ; Breast Neoplasms ; Breast ; Humans ; Lymphocytes, Tumor-Infiltrating ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Sexually Transmitted Diseases

OBJECTIVESTo study the factors influencing the non-sentinel lymph node(NSLN) status and to assess performance of Memorial Sloan-Kettering Cancer Center (MSKCC) nomogram in predicting sentinel lymph node(SLN) metastases in a SLN positive Chinese breast cancer population.

METHODSData were collected from breast cancer patients who were diagnosed with pathological positive sentinel lymph node and received further axillary lymph node dissection(ALND) in Shanghai Ruijin Hospital from January 2011 to August 2014. MSKCC nomogram was used to calculate each patient's NSLN metastasis risk score. The receiver operator characteristic curve (ROC curve) and the area under the ROC curve (AUC) was used to assess the predictive accuracy of the model.

RESULTSAmong the 1 147 patients who received sentinel biopsy, 150 SLN positive patients who received ALND were enrolled in this study. By univariate analysis, multifocal breast cancer (χ(2)=5.887, P=0.015), SLN+ /SLN ratio (χ(2)=6.683, P=0.010) and abnormal axillary lymph node displayed by ultrasound (χ(2)=7.736, P=0.005) were the influencing factors of NSLN metastases. By multivariate analysis, multifocal breast cancer (OR=7.25, 95% CI: 1.73 to 30.43, P=0.007), SLN+ /SLN ratio ≥ 0.5 (OR=2.564, 95% CI: 1.22 to 5.39, P=0.013) and abnormal axillary lymph node displayed by ultrasound (OR=2.471, 95% CI: 1.18 to 5.19, P=0.017) were the independent influencing factors of NSLN metastases. The AUC of MSKCC nomogram in this population was 0.677.

CONCLUSIONSFor breast cancer patients with positive sentinel lymph node, multifocality, SLN+ /SLN ratio and axillary lymphadenopathy displayed by ultrasound is related to NSLN metastasis. MSKCC has low accuracy in predicting NSLN status of this population.

Area Under Curve ; Axilla ; Breast Neoplasms ; China ; Humans ; Lymph Node Excision ; Lymph Nodes ; Lymphatic Metastasis ; Multivariate Analysis ; Nomograms ; ROC Curve ; Risk Factors ; Sentinel Lymph Node Biopsy

Objective We performed comprehensive proteomic analyses of synovial fluid by using the isobaric tags for relative and absolute quantitation (iTRAQ) method and LC-MS/MS, and searched for candidate biomarkers for osteoarthritis. Methods Synovial fluid was collected from patients with late-stage OA, earlystage OA and the control group. Molecular variations were detected by the iTRAQ method. T test was used for statistical analysis. Results Using the iTRAQ method, we identified 1 283 proteins from synovial fluid of patients with osteoarthritis, of which 268 proteins were not reported previously. There were 72 proteins upregulated and 249 proteins down-regulated between moderate OA group and controls. One hundred and twentyeight proteins were up-regulated and 141 proteins were down-regulated between severe OA group and controls.One hundred and ninety-two proteins were up-regulated and 76 proteins were down-regulated between severe OA group and moderate OA group. Eight proteins were found to be up-regulated in the three groups.Conclusion This is an in-depth analysis of the synovial fluid proteome from patients with osteoarthritis by iTRAQ method. The catalog of proteins generated in this study will further expand our knowledge regarding the pathophysiology of osteoarthritis and help us in identifying good biomarkers for early diagnosis.