[Risk factors of massive introperative blood loss in posterior spinal fusion for adolescent idiopathic scoliosis].

Objective: To investigate the risk factors and prognostic impact of massive introperative blood loss in posterior spinal fusion (PSF) surgery for adolescent idiopathic scoliosis (AIS). Methods: Clinical data were collected of 1 896 AIS patients who underwent PSF surgery under general anesthesia in Drum Tower Hospital Affiliated to Nanjing University Medical School from November 2010 to October 2019 and retrospectively analyzed. According to the volume of intraoperative blood loss, the patients were divided into the massive introperative blood loss group [estimated blood loss (EBL)/estimated blood volume (EBV)≥30%] and the non-massive introperative blood loss group (EBL/EBV<30%). The perioperative parameters between the two groups were compared, single factor analysis and multivariate logistic regression analysis was performed to identify independent risk factors related to massive introperative blood loss in PSF surgery. Results: A total of 1 896 AIS patients who underwent PSF surgery were included in the study. There were 298 males and 1 598 females, with an age of (14.5±1.7) years. Among them, 633 (33%) experienced massive intraoperative blood loss. The factors significantly related to the massive blood loss during PSF surgery in this study are: sex, body mass index(BMI), preoperative blood platelet count (PLT), prothrombin time, international normalized ratio(INR), preoperative Cobb angle, duration of operation, the number of fused levels, the number of screws, thoracoplasty, intraoperative use of tranexamic acid and dexmedetomidine; The independent factors included duration of operation longer than 4 hours(OR=4.311,P<0.001), the number of fused levels to be more than 10(OR=4.044,P<0.001), thoracoplasty (OR=2.174,P=0.019), BMI lower than 18.1 kg/m2(OR=2.094,P<0.001), preoperative PLT less than 186.5×109/L(OR=1.480,P=0.009), preoperative INR larger than 1 (OR=1.531,P=0.003) and preoperative Cobb angle larger than 53°(OR=1.306,P=0.036) ;Intraoperative use of tranexamic acid (OR=0.770, P=0.047) and dexmedetomidine (OR=0.653, P=0.008) are protective factors for massive intraoperative blood loss. In addition, in the massive intraoperative blood loss group, length of postoperative hospital stay (P<0.001), volume of postoperative incision drainage (P<0.001), postoperative allogeneic blood transfusion rate (22.7% vs 14.3%, P<0.001), incidence of postoperative hypoalbuminemia (90.3% vs 80.7%, P<0.001) and the number of rescue opioid analgesic requirements after surgery (P=0.006) were significantly higher than those in the non-massive introperative blood loss group. Conclusions: Longer operation duration, higher number of fusion levels, lower BMI, lower preoperative PLT, higher INR, larger preoperative Cobb angle and intraoperative thoraplasty are independent risk factors for massive intraoperative blood loss in AIS patients undergoing PSF surgery. Intraoperative use of tranexamic acid and dexmedetomidine can reduce the risk of massive blood loss in PSF surgery. Massive intraoperative blood loss significantly affects the patient's prognosis.

[Risk factors for postoperative intestinal obstruction in patients undergoing robot-assisted laparoscopic radical prostatectomy].

Objective: To analyze the risk factors of postoperative intestinal obstruction (POI) in patients undergoing robot-assisted laparoscopic radical prostatectomy (RARP). Methods: The clinical data of 573 patients receiving RARP from January to December 2019 in Nanjing Drum Tower Hospital were analyzed retrospectively. According to the occurrence of POI, the cases were divided into the occurrence group and the non-occurrence group. The clinical data of the two groups were compared and the risk factors of POI were investigated by multivariate logistic regression. Results: Forty-five of 573 patients (7.9%) had POI. Between the two groups, preoperative underlying diseases (cardiopathy, COPD, hypoalbuminemia), preoperative chemotherapy, preoperative WBC, operation time, blood loss, blood transfusion rate, postoperative early fever, length of stay were statistically significant (P<0.05). Multivariable logistic regression analysis showed that heart disease (OR=2.331, P=0.036), COPD (OR=4.285, P=0.001), hypoalbuminemia(OR=2.142, P=0.026), blood loss (≥4.26 ml/kg) (OR=2.388, P=0.010), operative time (≥225 min) (OR=4.200, P<0.001), and postoperative early fever (OR=2.773, P=0.004) were independent risk factors for POI after RARP. Conclusions: The incidence of POI following RARP is related to multiple perioperative factors. Improving the preoperative heart and lung function, correcting hypoalbuminemia, reducing intraoperative bleeding, shortening the operation time, and preventing early postoperative infection may be important measures to reduce the risk of POI in RARP patients.

CRKL promotes proliferation, migration, invasion of laryngeal squamous cell carcinoma.

OBJECTIVE: This study aimed to investigate the expression and role of CT10 regulated kinase like (CRKL) in human laryngeal squamous cell carcinoma (LSCC) progression. PATIENTS AND METHODS: Seventy-four laryngeal cancer cases were detected by the immunohistochemistry S-P method and the results were analyzed. RNA interference was used to downregulate the expression of CRKL in Hep-2 cells. The silencing efficiency was detected by real-time PCR and Western blotting. The cell proliferation, migration, and invasion after transfection were detected by MTT, wound healing assay, transwell invasion assay, and apoptosis assay. Western blot was conducted to determine the function of CRKL/epithelial-mesenchymal transition (EMT) signaling pathway. RESULTS: The expression of CRKL was higher in LSCC tissues. Patients with higher CRKL expression were correlated with lymph node metastasis and postoperative survival rates. CRKL promoted proliferation, migration, and invasion of Hep-2 cells in vitro. CONCLUSIONS: These findings suggested that CRKL gene silencing significantly inhibited the proliferation, migration, invasion, and EMT signaling pathway of Hep-2 cells. CRKL is considered to be a new target for the treatment of LSCC.

Research on the correlation of changes in plasma lncRNA MEG3 with change in inflammatory factors and prognosis in patients with traumatic brain injury.

OBJECTIVE: To study the correlation between the plasma long non-coding RNA (lncRNA) maternally expressed gene 3 (MEG3) and the levels of inflammatory cytokines in patients with traumatic brain injury (TBI) and to evaluate its prognosis to screen new biological targets for the diagnosis and treatment of TBI. PATIENTS AND METHODS: 40 patients with TBI (TBI group) and 40 healthy people (control group) were collected and venous blood was drawn. The plasma MEG3 in subjects was quantitatively analyzed via quantitative Polymerase Chain Reaction (qPCR). Moreover, the levels of inflammatory cytokines [tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), IL-6, and IL-8] in plasma in each group were detected via enzyme-linked immunosorbent assay (ELISA). Finally, the correlation analysis was performed for the MEG3 expression level and inflammatory cytokine levels in patients with TBI. Patients were divided into high-expression MEG3 group and low-expression MEG3 group, high-level inflammatory cytokine group and low-level inflammatory cytokine group according to the median, followed by prognosis evaluation. The MEG3 expression level in TBI group was significantly decreased compared to that in control group, and the levels of inflammatory cytokines in plasma, including TNF-α, IL-1ß, IL-6, and IL-8, were significantly higher than in control group. RESULTS: The results of the correlation analysis showed that the expression level of plasma MEG3 had a significantly negative correlation with the level of each inflammatory cytokine. The prognostic analysis revealed that the prognosis of patients with high MEG3 expression level and low inflammatory cytokine levels was good, while it was poor in patients with low MEG3 expression level and high inflammatory cytokine levels; the difference was significant. In patients with TBI, the expression level of plasma MEG3 is decreased, while the inflammatory cytokine levels are increased, and there is a significantly negative correlation between the two items. CONCLUSIONS: The prognosis of patients with high MEG3 expression level and low inflammatory cytokine levels is good so MEG3 and inflammatory cytokines can be used as biomarkers for diagnosis and treatment of TBI, improving the prognosis of patients.

Making the invisible visible: improving detectability of MRI-invisible residual cervical cancer after conisation by DCE-MRI.

AIM: To determine whether dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) quantitative parameters increase the detectability of MRI-invisible residual cervical cancer after conisation. MATERIALS AND METHODS: This retrospective study included 59 patients with MRI-invisible cervical cancer, but positive conisation pathology. Thirty-five patients were confirmed to have residual cervical cancer, and 24 patients showed non-residual cervical cancer. DCE-MRI quantitative parameters were calculated in the anterior or posterior cervix according to the conisation position. Receiver operating characteristic (ROC) analysis was used to find the threshold of DCE-MRI parameters in differentiate residual cervical cancer patients from non-residual cervical cancer patients after conisation. RESULTS: For patients with residual cervical cancer, the Ktrans and Ve values were significantly higher than in their counterparts with non-residual cervical cancer (0.610±0.395 versus 0.366±0.305/min, p=0.013; and 0.703±0.270 versus 0.540±0.280%, p=0.028; respectively). The Ktrans showed the highest area under the ROC curve (AUC) of 0.705 (p=0.004) with a sensitivity of 67.6% and specificity of 68%. CONCLUSION: DCE-MRI quantitative parameters increased the detectability of MRI-invisible residual cervical cancer after conisation.

Effect of RRS1 gene knockdown on BT549 cell line proliferation and apoptosis in breast cancer.

The RRS1 regulator of ribosome synthesis has recently been reported a new target gene linked to cancer development. This study therefore investigates RRS1effectsb on BT549 cell proliferation and apoptosis in breast cancer. Western blot (WB) and real - time quantitative PCR (qPCR) were used to detect the relative expression of RRS1 in breast cancer cells BT-549 and the normal HMEC mammary gland epithelial cells. BT-549 cells were cultured and infected with retroviruses and RRS1 expression was detected by qPCR and WB. The MTT assay, Caspase-3/7 and flow cytometry (FCM) then detected growth and apoptosis in the BT549 breast cancer BT cell. WB detected the expression of Bcl-2 and Bax genes related to apoptosis at the protein level, and MTT assay confirmed that RRS1 knockdown significantly decreased cell viability (p<0.05) and induced apoptosis which was rescued by shRNA-RRS1 expression. The amount of caspase-3 increased significantly and apoptosis was obvious. The apoptotic cells amount analyzed by FCM was significantly increased and RRS1 knockdown also decreased the expression of apoptosis related protein bcl-2 and simultaneously increased the expression of Bax (p<0.05). Finally, the RRS1 gene was highly expressed in breast cancer cell line BT549 and its knockdown significantly reduced proliferation and apoptosis in BT549 cell. These results suggest that RRS1 is a novel gene related to breast cancer and has an important role in breast cancer proliferation and apoptosis.

[Efficacy of coarctation resection and aortoplasty with autologous pulmonary artery patch strategy for treating coarctation of the aorta combined with hypoplastic aortic arch in infants].

Objective: To investigate the outcomes of coarctation resection and aortoplasty with autologous pulmonary artery patch for treating coarctation of the aorta combined with hypoplastic aortic arch in infants. Methods: Clinical data of 21 infants with coarctation of the aorta and hypoplastic aortic arch, who underwent coarctation resection and aortoplasty with autologous pulmonary artery patch in Fuwai hospital from January 2009 to June 2016 were retrospectively analyzed. The age of the patients was 4 (2, 5) months,and the body weight of the patients was (5.3±1.6) kg. The patients were followed up to observe the surgery effect. Results: No perioperative death and serious complications occurred. When the patients were discharged,the systolic blood pressure of the right upper limb was lower than the preoperative systolic blood pressure ((85.7±5.9) mmHg(1 mmHg=0.133 kPa) vs. (100.7±16.6) mmHg, P<0.001),and the systolic blood pressure of the right lower limb was higher than the preoperative systolic blood pressure ((98.7±13.3) mmHg vs. (85.6±20.8) mmHg, P<0.001). The pressure gradient of aortic coarctation detected by echocardiography was lower than the preoperative pressure gradient ((13.1±3.8) mmHg vs. (46.2±17.1) mmHg, P<0.001). No restenosis was detected by echocardiography at discharge. Follow-up data were obtained in 19 patients, and the follow-up time was 18 (8, 45) months.The patients grew well, and no death occurred. Restenosis occurred in 3 cases, 1 patient underwent aortic balloon dilatation and the remaining 2 patients were under follow up observation. Computed tomography angiography showed that the morphology of aortic arch was normal without signs of aortic aneurysm. Conclusion: Coarctation resection with autologous pulmonary artery patch aortoplastystrategy is considered as a safe and effective surgical method for management of infant coarctation with hypoplastic aortic arch, and this surgery method is related with satisfactory early and mid-term outcomes in this patient cohort.

[Risk factors associated with massive drainage after posterior spinal orthopaedic surgery for adolescent scoliosis].

Objective: To identify risk factors that influence the massive drainage after posterior spinal orthopaedic surgery for adolescent scoliosis. Methods: A total of 1 461 patients from 11 to 18 years old diagnosed with adolescent scoliosis who underwent first posterior spinal orthopaedic surgery in affiliated Drum Tower Hospital, Medical School of Nanjing University between November 2010 and October 2015 were retrospectively reviewed. Patients were categorized on the basis of massive or normal drainage, with the boundary 30(th) percentile of drainage/estimated blood volume. Preoperative factors including age, gender, body mass index(BMI), ASA physical status, diagnostic type of scoliosis, main Cobb angle, laboratory tests, intraoperative factors including the number of fusion level and screws, tranexamic acid used or not, use of osteotomy and thoracoplasty, use of cell salvage technology, duration of operation, the volume of urine output, blood loss, fluid therapy and transfusion, postoperative factors including the length of hospital stay, number of transfusion, the volume of drainage, time of drain were collected. Univariate and multivariate analyses were used to determine risk factors which were independently associated with massive drainage. Results: The average drainage was (856.3±333.4)ml. 479(32.8%) patients had massive drainage(drainage≥30% of drainage/estimated blood volume). Multivariate analysis identified risk factors of massive drainage: BMI<17.63 kg/m(2), odds ratio(OR)=2.90, preoperative platelet count<190×10(9)/L (OR=1.67), preoperative main Cobb angle≥55 degrees(OR=1.66), number of fusion levels≥11(OR=2.33), number of screws≥15(OR=1.73), use of osteotomy(OR=1.54), intraoperative volume of crystalloids≥35.63 ml/kg(OR=1.40), intraoperative volume of colloids≥28.92 ml/kg(OR=1.82), intraoperative volume of transfusion≥19.55 ml/kg(OR=1.72), while the use of tranexamic acid(OR=0.26) was the only protective factor. Conclusion: BMI<17.63 kg/m(2,) preoperative platelet count<190×10(9)/L, preoperative main Cobb angle≥55 degrees, number of fusion levels≥11, number of screws≥15, use of osteotomy, intraoperative volume of crystalloids≥35.63 ml/kg, intraoperative volume of colloids≥28.92 ml/kg, intraoperative volume of transfusion≥19.55 ml/kg were risk factors associated with massive drainage after posterior spinal orthopaedic surgery for adolescent scoliosis, while the use of tranexamic acid could decrease the possibility of massive drainage.

MiR-145 silencing promotes steroid-induced avascular necrosis of the femoral head repair via upregulating VEGF.

OBJECTIVE: To investigate the role of miR-145 silencing in the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), B-cell lymphoma-2 (Bcl-2), BCL2-Associated X(bax) and caspase-3 in avascular necrosis of femoral head (ANFH). MATERIALS AND METHODS: A total of 12 healthy wild-types (the control group) and 12 miR-145 knock-out (miR-145-/-) (the experimental group) adult New Zealand white rabbits were selected to construct ANFH model with a steroid. Four weeks later, immunohistochemistry, qRT-PCR and Western blot were performed to measure the VEGF, bFGF, Bcl-2, bax, caspase-3, ß-catenin as well as c-Myc expression. Terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) staining analysis was used to detect the apoptosis of bone cells in each group. RESULTS: Compared with the control group, the expression of VEGF, bFGF, Bcl-2, ß-catenin and c-Myc in the miR-145-/- group raised (p<0.05). Moreover, the expression level of bax and caspase-3 significantly decreased in the miR-145-/- group (p<0.05). TUNEL staining showed decreased apoptosis in the miR-145-/- group. CONCLUSIONS: MiR-145 silencing promotes bone repair of ANFH via upregulating VEGF, bFGF and inhibiting the bone cells apoptosis through Wnt/ß-catenin pathway.

MicroRNA-34a/EGFR axis plays pivotal roles in lung tumorigenesis.

MicroRNAs (miRNAs) are vital in the regulation of tumor progression and invasion. Dysregulation of miRNAs has been linked to the development of various types of human cancers, including non-small-cell lung cancer (NSCLC). However, the effect of miRNA-34a (miR-34a), a key regulator of tumor suppression, on the tumorigenesis of NSCLC has not been fully elaborated. Herein, we reveal that miR-34a is significantly downregulated in NSCLC tissues and cell lines, suggesting that miR-34a might function as a tumor suppressor in lung cancer. We also confirmed that epidermal growth factor receptor (EGFR) is a direct target of miR-34a, and our data reveal that siRNA knockdown of EGFR can inhibit cell proliferation, promote apoptosis and arrest cell-cycle progression. In addition, EGFR can reverse the suppressive function of miR-34a overexpression on proliferation and cell apoptosis. Furthermore, in vivo experiments demonstrated that miR-34a suppress tumor growth, both in the A549 xenograft model, as well as in the metastatic tumors in nude mice. Taken together, our findings suggest that miR-34a inhibits NSCLC tumor growth and metastasis through targeting EGFR.

Protective Effect of Edaravone Against Aß25-35-Induced Mitochondrial Oxidative Damage in SH-SY5Y Cells.

Amyloid-ß (Aß)-induced oxidative stress plays an important role in the pathogenesis of Alzheimer's disease (AD). Recent studies show that Aß accumulation may lead to mitochondrial oxidative damage. In the present study, we investigated the protective effect of edaravone on mitochondrial damage in SH-SY5Y cells treated with Aß25-35. SH-SY5Y cells were pre-treated with 20, 40 or 80 µM edaravone before treatment with 25 µM Aß25-35. After 24h cell culture, cellular apoptosis, intracellular reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨm), ATP levels and mitochondrial morphology were evaluated. SH-SY5Y cells exposed to Aß25-35 had high levels of apoptosis and ROS; loss of ΔΨm, decreased ATP levels and presence of mitochondrial swelling. However, these effects were significantly inhibited by edaravone pre-treatment. These results indicate that edaravone prevents mitochondria oxidative damage caused by Aß in SH-SY5Y cells, which suggests that it may have potential clinical application in AD therapy.

USP4 promotes invasion of breast cancer cells via Relaxin/TGF-ß1/Smad2/MMP-9 signal.

OBJECTIVE: Ubiquitin-specific protease 4 (USP4) is a deubiquitinating enzyme with key roles in the regulation of TGF-ß1 signaling, suggesting its importance in tumorigenesis. However, the underlying mechanisms causing this are not entirely clear. In the present study, we investigated the effect of USP4 on invasion and tumorigenesis of breast cancer cells, and explored its mechanism. MATERIALS AND METHODS: Effects of USP4 overexpression or USP4 silencing by small interfering RNA (USP4 siRNA) on invasion of breast cancer MDA-MB-231 and T47D cells in vitro was detected. Using siRNAs and inhibitors to examine the USP4 signaling pathway. RESULTS: The migration and invasion assays showed that USP4 promotes human breast cancer cell migration and invasion by USP4 overexpression, and knockdown of USP4 by siRNA inhibits human breast cancer cell migration and invasion. Treatment with RLX siRNAs, TGF-ß1 siRNAs, Smad2 siRNAs or BB94 (MMPs inhibitor) to USP4-overexpressing breast cancer cells revealed that USP4- induced RLX via TGF-ß1 pathway promotes the cell migration and invasion. Further studies demonstrated that USP4-mediated TGF-ß1 activation not only enhances the phosphorylation of Smad2 through TGF-ß, but also directly upregulate matrix metalloproteinase (MMP)-9-mediated cell migration and invasion of breast cancer cells. CONCLUSIONS: Therapies targeting the USP4 inhibits invasion of breast cancer cells via Relaxin/TGF-ß1/Smad2/MMP-9 signal. These results indicate that USP4 is an attractive target for breast cancer therapy.

Knockdown of CD24 inhibits proliferation, invasion and sensitizes breast cancer MCF-7 cells to tamoxifen in vitro.

OBJECTIVE: CD24 is overexpressed in breast cancer, and patients with high CD24 expression was resistant to tamoxifen treatment. Furthermore, treatment with CD24 antibody to inhibit CD24 expression could induce apoptosis and inhibits migration in breast cancer cells in vitro. In this study, we investigated the anti-tumor efficacy of CD24 knockdown using siRNA targeting CD24 on proliferation, invasion and sensitivity to tamoxifen (TAM) of breast cancer MCF-7 cells in vitro. MATERIALS AND METHODS: CD24 siRNA vector (CD24-siRNA) and empty plasmid vector (EP) were transiently transfected into the breast cancer MCF-7 cells and the knockdown efficacy was assessed by Western blot analysis. The effects of CD24 knockdown on cell viability, apoptosis and sensitivity to TAM in MCF-7 cells were determined using methyl thiazolyl blue tetrazolium bromide (MTT), ELISA and terminal deoxynucleotidyl transferase-mediated nick end-labeling (TUNEL) assays. The effects of CD24 knockdown on cell invasion and migration were determined using chemoinvasion assay and wound scratch assay, respectively. RESULTS: Transfection of CD24-siRNA effectively down-regulated CD24 expression in MCF-7 cells in vitro. CD24 suppressed showed significantly decreased proliferation, invasion and increased apoptosis as well as increased sensitivity to TAM in vitro in MCF-7 cells. CONCLUSIONS: Knockdown of CD24 expression by CD24-siRNA significantly inhibited invasion and cell viability, and induced apoptosis and increased sensitivity of MCF-7 cells to TAM, indicating that knockdown of CD24 by siRNA might be a potential therapeutic approach against human breast cancer.

Clinical, demographic and histopathological features of symmetrical acral keratoderma.

BACKGROUND: A series of cases of symmetrical acral keratoderma have been described recently in China. However, no studies about its demographic information and epidermal barrier function have been documented. OBJECTIVES: To describe the clinical manifestation, demographic information and clinicopathological features of 71 cases with symmetrical acral keratoderma. PATIENTS AND METHODS: Seventy-one cases with symmetrical acral keratoderma were retrospectively reviewed. Their demographic information, clinical manifestations, histopathology and epidermal barrier function were analysed. RESULTS: Among these patients, there were 64 males and seven females, ranging in age from 4 to 53 years with an average age at onset of 27 ± 8·9 years. Clinical manifestation was characterized by brown hyperkeratotic patches over the dorsum of the hands, palms and feet, dorsal digits and wrists, elbows, knees and ankles. The lesions became dramatically whitish with mild swelling immediately after soaking in water and resolved spontaneously in winter. In patients, a moderate increase of transepidermal water loss (TEWL) from 16·16 ± 6·15 to 9·9 ± 4·21 g m(-2)  h(-1) (P = 0·0054) and a moderate decrease of skin hydration from 65·9 ± 5·06 to 42·58 ± 10·73 (P < 0·01) in comparison with the control group were observed. Histopathological examination revealed epidermal hyperkeratosis, acanthosis and papillomatous hyperplasia as well as dermal infiltration with a few lymphocytes. CONCLUSIONS: Symmetrical acral keratoderma is characterized by symmetry, acra, keratinization and marked seasonal changes. The epidermal barrier function of the skin was negatively affected.

Collisional 3He and 129Xe frequency shifts in Rb-noble-gas mixtures.

The Fermi-contact interaction that characterizes collisional spin exchange of a noble gas with an alkali-metal vapor also gives rise to NMR and EPR frequency shifts of the noble-gas nucleus and the alkali-metal atom, respectively. We have measured the enhancement factor κ0 that characterizes these shifts for Rb-129Xe to be 493±31, making use of the previously measured value of κ0 for Rb-3He. This result allows accurate 129Xe polarimetry with no need to reference a thermal-equilibrium NMR signal.