Endothelial extracellular vesicles induce acute lung injury via follistatin-like protein 1.

Cardiopulmonary bypass has been speculated to elicit systemic inflammation to initiate acute lung injury (ALI), including acute respiratory distress syndrome (ARDS), in patients after cardiac surgery. We previously found that post-operative patients showed an increase in endothelial cell-derived extracellular vesicles (eEVs) with components of coagulation and acute inflammatory responses. However, the mechanism underlying the onset of ALI owing to the release of eEVs after cardiopulmonary bypass, remains unclear. Plasma plasminogen-activated inhibitor-1 (PAI-1) and eEV levels were measured in patients with cardiopulmonary bypass. Endothelial cells and mice (C57BL/6, Toll-like receptor 4 knockout (TLR4-/-) and inducible nitric oxide synthase knockout (iNOS-/-)) were challenged with eEVs isolated from PAI-1-stimulated endothelial cells. Plasma PAI-1 and eEVs were remarkably enhanced after cardiopulmonary bypass. Plasma PAI-1 elevation was positively correlated with the increase in eEVs. The increase in plasma PAI-1 and eEV levels was associated with post-operative ARDS. The eEVs derived from PAI-1-stimulated endothelial cells could recognize TLR4 to stimulate a downstream signaling cascade identified as the Janus kinase 2/3 (JAK2/3)-signal transducer and activator of transcription 3 (STAT3)-interferon regulatory factor 1 (IRF-1) pathway, along with iNOS induction, and cytokine/chemokine production in vascular endothelial cells and C57BL/6 mice, ultimately contributing to ALI. ALI could be attenuated by JAK2/3 or STAT3 inhibitors (AG490 or S3I-201, respectively), and was relieved in TLR4-/- and iNOS-/- mice. eEVs activate the TLR4/JAK3/STAT3/IRF-1 signaling pathway to induce ALI/ARDS by delivering follistatin-like protein 1 (FSTL1), and FSTL1 knockdown in eEVs alleviates eEV-induced ALI/ARDS. Our data thus demonstrate that cardiopulmonary bypass may increase plasma PAI-1 levels to induce FSTL1-enriched eEVs, which target the TLR4-mediated JAK2/3/STAT3/IRF-1 signaling cascade and form a positive feedback loop, leading to ALI/ARDS after cardiac surgery. Our findings provide new insight into the molecular mechanisms and therapeutic targets for ALI/ARDS after cardiac surgery.

LncRNA IGBP1-AS1 targets miR-150-5p to increase ZEB1 expression in nasopharyngeal carcinoma.

Background: Nasopharyngeal carcinoma (NPC), which is a form of cancer arising from the epithelium of the nasopharynx, remains highly prevalent, particularly in Southeast Asia and Southern China. Dysregulated long non-coding RNAs (lncRNAs) are involved in several malignancies, including the growth and aggression of tumors. LncRNA IGBP1-AS1 plays an important role in the advancement of breast cancer, but its role in NPC has not yet been explored. Methods: LncRNA IGBP1-AS1 levels in human NPC samples compared with their matched adjacent normal tissues by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The proliferation alteration of NPC cells was tested in vitro. Luciferase reporter assay and RNA immunoprecipitation (RIP) were carried out to reveal the interaction between lncRNA IGBP1-AS1, miR-150-5p, and zinc finger e-box binding homeobox 1 (ZEB1). Results: In the study, we found that IGBP1-AS1 was found to be significantly upregulated in the NPC samples and cell lines. The knockdown of IGBP1-AS1 impeded in-vitro proliferation of the NPC cells. Bioinformatics and reporter assays revealed an association between IGBP1-AS1 and miR-150-5p. The expression of ZEB1 was increased by the microRNA (miRNA) sequestering in human NPC. Conclusions: The progression of NPC tumors is facilitated by lncRNA IGBP1-AS1 via miR-150-5p and regulates ZEB1 expression.

[Two Serum Free Light Chain Detection Systems in the Diagnosis of Multiple Myeloma].

OBJECTIVE: To investigate the comparability of the Freelite, Binding Site, Beckman and N Latex FLC, Siemens in the detection of serum free light chain (sFLC) . METHODS: Fifty newly diagnosed multiple myeloma (MM) patients in Tianjin Institute of Blood Research from November 2019 to February 2020 were enrolled. The two systems (Freelite, Binding Site, Beckman and N Latex FLC, Siemens) were used to detect the sFLC of the samples. Outlier detection was performed by ESD method, methodological comparison and deviation assessment were performed by Passing-Bablok regression and Bland-Altman regression. RESULTS: Both the systems could quantitatively analyze free kappa light chain serum samples and free lambda light chain samples. Freelite, Binding Site, Beckman and N Latex FLC, Siemens free light chain test showed FLC-κ：36.5 (6.5, 194), 40.5 (6.94, 288), FLC-λ: 30.1 (4.3, 170.5), 35.1 (2.28, 526), rFLC (FLC-κ/ FLC-λ) : 0.82 (0.05, 43.25), 1.03 (0.03, 32.04), dFLC (｜FLC-κ- FLC-λ｜) : -5.8 (-161.97, 183.7), 1.1 (-505.1, 279.01), which existed no outliers. There were systematic differences, and the deviation level was not within the clinically acceptable range. CONCLUSION: Both the systems can meet the needs of clinical diagnosis and treatment, but there is a significant deviation between the two systems, the results are not comparable, and should be analyzed separately. In particular, the same system should be selected for monitoring the prognosis of MM.

Effects of glenohumeral internal rotation deficit on baseball pitching among pitchers of different ages.

BACKGROUND: Previous studies have reported that glenohumeral internal rotation deficit (GIRD) may increase the risk of shoulder injury. However, the effects of GIRD on baseball pitching among pitchers of different age groups are still unclear. METHODS: The study participants were 24 high school and 24 university pitchers. For each age group, the pitchers were evenly divided into a GIRD group and a normal group. The pitching motion of each participant was captured using a motion analysis system at a sampling frequency of 300 Hz. The kinematics and kinetics of the throwing shoulder and trunk were quantified, and statistical differences between the groups were examined by 2-sample t tests. RESULTS: For both age groups, significant differences were observed in shoulder external rotations of the GIRD and normal groups. Compared with the university pitchers in the normal group, the university pitchers with GIRD exhibited a greater shoulder loading and did more internal rotation work in the acceleration phase. The high school pitchers with GIRD showed a larger trunk tilt and less trunk rotation than the university pitchers with GIRD. However, the university pitchers with GIRD exhibited a larger shoulder posterior force and horizontal adduction torque than the high school pitchers with GIRD. CONCLUSION: Pitchers with GIRD do change their pitching motions, and the greater resulting shoulder joint loading predisposes them to a greater risk of shoulder injury, especially among university pitchers.

Endothelial microparticles are increased in congenital heart diseases and contribute to endothelial dysfunction.

BACKGROUND: We previously demonstrated that endothelial microparticles (EMPs) are increased in mitral valve diseases and impair valvular endothelial cell function. Perioperative systemic inflammation is an important risk factor and complication of cardiac surgery. In this study, we investigate whether EMPs increase in congenital heart diseases to promote inflammation and endothelial dysfunction. METHODS: The level of plasma EMPs in 20 patients with atrial septal defect (ASD), 23 patients with ventricular septal defect (VSD), and 30 healthy subjects were analyzed by flow cytometry. EMPs generated from human umbilical vascular endothelial cells (HUVECs) were injected into C57BL6 mice, or cultured with HUVECs without or with siRNAs targeting P38 MAPK. The expression and/or phosphorylation of endothelial nitric oxide synthase (eNOS), P38 MAPK, and caveolin-1 in mouse heart and/or in cultured HUVECs were determined. We evaluated generation of nitric oxide (NO) in mouse hearts, and levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in cultured HUVECs and in mice. RESULTS: EMPs were significantly elevated in patients with ASD and VSD, especially in those with pulmonary hypertension when compared with controls. EMPs increased caveolin-1 expression and P38 MAPK phosphorylation and decreased eNOS phosphorylation and NO production in mouse hearts. EMPs stimulated P38 MAPK expression, TNF-α and IL-6 production, which were all inhibited by siRNAs targeting P38 MAPK in cultured HUVECs. CONCLUSIONS: EMPs were increased in adult patients with congenital heart diseases and may contribute to increased inflammation leading to endothelial dysfunction via P38 MAPK-dependent pathways. This novel data provides a potential therapeutic target to address important complications of surgery of congenial heart disease.

[Safety and efficacy of subcutaneous administration of bortezomib in the treatment of multiple myeloma].

OBJECTIVE: To analyze the efficacy and safety of subcutaneous administration of bortezomib in the treatment of multiple myeloma (MM) patients. METHODS: A total of 26 MM patients were enrolled in this study and treated with BDT (bortezomib-dexamethasone-thalidomide). In the 26 MM patients, 12 patients received subcutaneous administration of Bortezomib while 14 patients received conventional intravenous administration. The outcomes and adverse effects of two groups were retrospectively evaluated and compared. RESULTS: Overall response (OR) rates in the two groups were 75.00% and 71.43% respectively, in which complete remission (CR) plus very good complete remission (VGPR) rates were 50.00% and 47.14%, while CR rates were 16.67% and 28.57%. There were no statistically significant difference (P > 0.05). Time to achieve effectiveness in two groups was similar (P > 0.05). More than half patients in both groups achieved partial remission after the first treatment course and CR after the fourth course. Compared to the intravenous group, peripheral neuropathy rates remained significantly lower in subcutaneous group (16.67% vs. 64.29%, P = 0.021). The intravenous group had 7.14% grade 3 or worse, peripheral neuropathy but none found in the subcutaneous group. Rash occurred only in subcutaneous group (66.67%), but it was local, mild and transitional. No significant differences of other adverse events between the two groups were observed. CONCLUSION: Subcutaneous administration of bortezomib offers similar efficacy to standard intravenous administration in the treatment of multiple myelom, with an improved safety for lower rate of peripheral neuropathy.

[Influence of obstructive jaundice on postoperative complications and mortality after pancreaticoduodenectomy: analysis of the 25-year single-center data].

OBJECTIVE: To study the influence of the depth of jaundice, the duration of jaundice and preoperative biliary drainage (PBD) on postoperative complications and mortality after pancreaticoduodenectomy (PD). METHODS: A retrospective review was performed of the medical records of 1025 patients who underwent PD between June 1986 and December 2010. The patients comprised 659 men and 366 women, ranging from 4 to 81 years old with a mean age of (54 ± 12) years. The indications for PD were malignant disease in 869 patients (84.78%) and benign or borderline tumors in 156 patients (15.22%). The operative procedures performed were pylorus-preserving modification in 279 patients and conventional PD, i.e. Whipple's operation in 746 patients. Complications after PD were compared among the different groups which was classified according to the depth of obstructive jaundice, the duration of obstructive jaundice and whether undergoing preoperative biliary drain or not, and the analysis was made by variance analysis and χ(2) test respectively. RESULTS: The depth of jaundice did not significantly affect the incidence of complications after PD except for the hemorrhage complication (χ(2) = 11.06, P = 0.03). The duration of jaundice had no much influence on the postoperative complications and mortality. PBD could not reduce the postoperative complications and mortality, however, it would increase the incidence of postoperative incision infection (χ(2) = 9.84, P = 0.01). No significant relationship was observed between the duration of PBD and the postoperative complications and mortality. CONCLUSIONS: Either the depth or duration of obstructive jaundice has no relationship with the postoperative complications and mortality after PD but the postoperative hemorrhage. Patients undergoing PD can not be benefited from PBD. Consequently, PBD should not be performed routinely, but it can be used in some serious patients with severe depth of jaundice who can not received surgery at once.

Prognostic value of the multidrug resistance transporter ABCG2 gene polymorphisms in Chinese patients with de novo acute leukaemia.

BACKGROUND: Functional polymorphisms of the ABCG2 gene may contribute to individual variability in drug response and the prognosis of patients. METHODS: In the present study, the genetic polymorphisms and expression of ABCG2 were analysed in blasts cells obtained from 184 Chinese patients with de novo acute leukaemia to investigate their possible association with clinical outcomes. RESULTS: A novel synonymous ABCG2-single nucleotide polymorphism (SNP) at exon 16 (13561218 C/T) and five known SNPs at exon 2 (13608835 G/A), exon 5 (13600044 C/A), intron 10 (13576005 C/T), intron 13 (13564503 C/T) and intron 14 (13563578 A/G) were identified with occurrence rates of 1.1%, 64.1%, 30.4%, 21.2%, 39.7% and 28.8%, respectively. We found that patients with the ABCG2 34GG genotype displayed longer disease free survival (DFS) (P<0.001) and overall survival (OS) (P<0.001) than those with the 34GA/AA genotypes. Furthermore, the DFS and OS were significantly diminished in bone marrow transplantation (BMT) patients with the 34GA/AA genotypes relative to those with the 34GG genotype. CONCLUSIONS: These results suggest that these highly prevalent ABCG2 34GA/AA genotypes are associated with poor prognosis of Chinese patients with acute leukaemia and BMT patients.

Up-regulation of ABCB1/P-glycoprotein by escaping promoter hypermethylation indicates poor prognosis in hematologic malignancy patients with and without bone marrow transplantation.

We investigated the correlation between MDR1 promoter methylation status and MDR1 expression in 228 hematologic malignancies patients and 90 healthy controls. High level of MDR1 mRNA correlated to promoter hypomethylation and strongly associated with poor prognosis indicated by 2-year survival rates, poor CR rate (without BMT) and high relapse rate (with BMT). Furthermore, relative luciferase activity of methylated MDR1 at promoter -50 region was significantly higher than that of the unmethylated. In addition, MDR1 in K562 cells elevated significantly after 5-Aza-dC treatment. In summary, MDR1 promoter hypomethylation conferred its up-regulation and indicated poor prognosis in patients with and without BMT.

[Mechanism of endogenous carbon monoxide effect on hydrogen sulfide in guinea pigs with established allergic rhinitis].

OBJECTIVE: To study the effect of carbon monoxide (CO) on hydrogen sulfide (H2S) in guinea pigs with allergic rhinitis (AR) through intervention treatment. METHODS: AR model in guinea pigs was established by using ovalbumin. The animals were divided into three groups. Group one was sensitized continuously by ovalbumin, group two was treated with Hemin as induction group, and group three was treated with zinc protoporphyrin (ZnPP) as suppression group. The guinea pigs treated with saline were used as control. The behavior science scores, eotaxin concentration of nasal lavage, IgE in blood serum were recorded, and the plasma concentrations of CO and H2S were determined, then the expression of hemeoxygenase (HO)-1, cystathionine-beta-synthase (CBS) and cystathionine-gamma-lyase (CSE) were measured in nasal mucosa by fluorescent quantitative RT-PCR. RESULTS: The behavior science scores, concentration of eotaxin in nasal lavage, IgE in blood serum and concentration of CO in plasma of sensitized group were higher than those of control (P<0.01), and the expression of HO-1 in nasal mucosa was also higher than control [(7.61+/-2.80)x10(-3) vs (2.32+/-1.14)x10(-3), P<0.05]. All these items were higher when treated with Hemin and lower when treated with ZnPP (P<0.05). The concentration of H2S in plasma was lower than control with significant differences [(14.80+/-1.60) micromol/L vs (18.90+/-1.00) micromol/L, P<0.01], the expression of CSE was also lower than control (P<0.05), and both of them were lower with Hemin induced and higher with ZnPP (P<0.05). The expression of CBS was very low and had no significant differences between groups (P>0.05), so it indicated that the CSE was the key enzyme for endogenous H2S product in nasal mucosa. Moreover the concentration of H2S was negatively correlated with CO (r=-0.702, P<0.001). CONCLUSIONS: Endogenous CO and H2S play a significant role in the pathogenesis of AR, and HO-1 and CSE are the main speed-relate enzymes respectively. The H2S is also influenced by CO.

Sensitization of ABCG2-overexpressing cells to conventional chemotherapeutic agent by sunitinib was associated with inhibiting the function of ABCG2.

Sunitinib is an ATP-competitive multi-targeted tyrosine kinase inhibitor. In this study, we evaluated the possible interaction of sunitinib with P-glycoprotein (P-gp, ABCB1), multidrug resistance protein 1 (MRP1, ABCC1), breast cancer resistance protein (BCRP, ABCG2) and lung-resistance protein (LRP) in vitro. Our results showed that sunitinib completely reverse drug resistance mediated by ABCG2 at a non-toxic concentration of 2.5muM and has no significant reversal effect on ABCB1-, ABCC1- and LRP-mediated drug resistance, although a small synergetic effect was observed in combining sunitinib and conventional chemotherapeutic agents in ABCB1 overexpressing MCF-7/adr and parental sensitive MCF-7 cells, ABCC1 overexpressing C-A120 and parental sensitive KB-3-1 cells. Sunitinib significantly increased intracellular accumulation of rhodamine 123 and doxorubicin and remarkably inhibited the efflux of rhodamine 123 and methotrexate by ABCG2 in ABCG2-overexpressing cells, and also profoundly inhibited the transport of [(3)H]-methotrexate by ABCG2. However, sunitinib did not affect the expression of ABCG2 at mRNA or protein levels. In addition, sunitinib did not block the phosphorylation of Akt and Erk1/2 in ABCG2-overexpressing or parental sensitive cells. Overall, we conclude that sunitinib reverses ABCG2-mediated MDR through inhibiting the drug efflux function of ABCG2. These findings may be useful for cancer combinational therapy with sunitinib in the clinic.

[Influence of carbon monoxide on the expression of inducible nitric oxide synthase mRNA in guinea pigs with allergic rhinitis.].

OBJECTIVE: To study the impact of carbon monoxide (CO) on expression levels of inducible nitric oxide synthase (iNOS) mRNA in guinea pigs with allergic rhinitis (AR). METHODS: Twenty four guinea pigs were divided randomly into four study groups with 6 guinea pigs in each. The guinea pigs in the first group were treated with saline only (Group 1, the healthy controls). The remaing guinea pigs were sensitized by ovalbumin and thus establishing the AR models. After sensitization, the animals in the second group remained untreated (Group 2, AR control group). The third group was treated with Hemin as the induction group, and the fourth group was treated with Zinc protoporphyrin (ZnPP) as the suppression group. The plasma concentration of carboxyhemoglobin (COHb) was measured, which represents the concentration of CO. The expression levels of Heme oxygenase-1 (HO-1) and NOS mRNAs in nasal mucosa were determined by fluorescent quantitative RT-PCR. RESULTS: AR models were established successfully in all study guinea pigs. The concentrations of COHb (x(-) +/- s) in plasma of the second group (2.27% +/- 1.13%) were significantly (q = 4.10, P < 0.01) higher than those of healthy controls (1.08% +/- 0.24%). The plasma concentration of COHb in the third group (3.17% +/- 0.68%) were also significantly higher (q = 3.12, P < 0.05) than those in the second group. The expression levels of HO-1 and iNOS in nasal mucosa of the second group [(7.80 +/- 1.60) x 10(-3) and (5.81 +/- 0.05) x 10(-3), respectively] were also significantly (q equals 5.52 and 7.21, respectively, P < 0.01) higher than those of controls [(1.96 +/- 0.71) x 10(-3) and (0.97 +/- 0.05) x 10(-3), respectively]. The expression levels of HO-1 and iNOS in the nasal mucosa of the third group [(11.89 +/- 4.78) x 10(-3) and (7.42 +/- 0.70) x 10(-3), respectively] were significantly (q equals 3.86 and 2.22, P < 0.05) higher than those of the second group. The expression levels of HO-1 and iNOS in nasal mucosa of the fourth group [(3.82 +/- 0.98) x 10(-3) and (2.34 +/- 0.04) x 10(-3), respectively] were significantly (q equals 3.76 and 5.18, P < 0.05) lower than those in the second group. CONCLUSIONS: Endogenous carbon monoxide influenced the expression levels of iNOS in nasal mocusa in guinea pigs with AR.

[Treatment of the windblow hand deformity].

OBJECTIVE: To discuss the treatment of the windblow hand deformity. METHODS: We treat 18 patients with operations step by step. First phase: to improve the thumb function; Second phase: shift the flexor digitorum superficialis of the middle and little finger to improve the ulnar drift of the digits; Third phase : to solve the flection deformity of the palm and digits. RESULTS: We follow the patients for 6 - 72 months, only lost 3.11 of 15 patients have the thumb function recovered. 9 patients had their ulnar drift of the digits corrected, 2 patients partly corrected, 4 have relapse. 5 among the 15 patients were offered second operation, to have the ulnar arthrosis bursa contracted, using abduction splint, after rehabilitation 4 patients have the deformity corrected, 1 has partly corrected. CONCLUSIONS: Operation step by step is better for the complex windblow hand deformity, solve one major problem each time,and systemic rehabilitation therapy is needed for satisfied curative effect.

[To inquiry into clinical-pathological factors influencing cytological accuracy of pancreatic head lesions].

OBJECTIVE: To inquiry into clinical-pathological factors influencing cytological accuracy of pancreatic head lesions. METHODS: Cytology was retrospectively evaluated in 94 inpatients with a mass in head of pancreas existing with chronic pancreatitis in the past decade, the results of cytology were compared with clinical pathology or clinical follow-up to estimate the value and accuracy of cytology in detecting pancreatic cancer. Pancreatic clinical-pathology includes size of mass and component of mass which was composed of cancerous mode of development including shape of conglomeration, nest and pervasion, and pancreatic ductal epithelium inside the vicinity of 1 cm around the mass. Pancreatic ductal epithelium were divided into PanIN1, PanIN2 and PanIN3 three types according to classified criterion of pancreatic intraepithelial neoplasia (PanIN). RESULT: Forty-six patients were pathologically diagnosed as pancreatic cancer and five patients as chronic pancreatitis, accordingly, 29 malignant, 5 suspicious, 10 atypical hyperplasia among of them 5 malignant, 3 hyperplastic ductal epithelium, 1 nondiagnostic results due to interfered by blood and 3 insufficient specimens. 43 patients were clinically diagnosed as chronic pancreatitis. Cytologic evaluation of pancreatic cancer has an 84.2% accuracy less than or equal to 2.5 cm and 71.9% larger than 2.5 cm. Shape of conglomeration, and nest have more accurate than pervasion in cytological diagnosis. CONCLUSIONS: Cancerous mode of development is a vital factor influencing accuracy of cytology, cytological estimation of atypical hyperplasia and is still waiting for further investigation.

Protective effects of lidocaine injected into the hepatoduodenal ligament on warm ischemia-reperfusion injury to the rat liver.

BACKGROUND: Ischemia-reperfusion (IR) injury to the liver is still a critical and daunting problem in the field of hepatobiliary surgery. Ischemic preconditioning (IP) of the liver serves as an effective approach against IR injury. This study was to develop a novel procedure that could mimic IP, but might be more feasible than IP during surgery. METHODS: Eighty-two SD rats were randomly divided into 5 groups. L group (n = 21): 0.4% lidocaine (10 mg/kg) was injected into the hepatoduodenal ligament 10 minutes before a 40-minute hepatic IR. IP group (n = 16): a 5-minute ischemia was followed by a 10-minute reperfusion prior to a 40-minute hepatic IR. ILR group (n = 15): after a 40-minute ischemia of the liver, 0.4% lidocaine (10 mg/kg) was injected into the hepatoduodenal ligament 10 minutes prior to a 40-minute reperfusion of the liver. IR group (n = 15): the liver of the rat was subjected to a 40-minute IR. Control group (n = 15): 0.9% sodium chloride was injected into the hepatoduodenal ligament without other treatments. The levels of plasma alanine transaminase (ALT) and aspartate transaminase (AST) were determined for each group after treatment. RESULTS: The mean concentrations of ALT and AST were (379.80 +/- 141.69) U/L and (606.05 +/- 220.26) U/L for the L group, (334.64 +/- 141.94) U/L and (625.68 +/- 267.06) U/L for the IP group, (523.36 +/- 170.35) U/L and (765.47 +/- 238.45) U/L for the ILP group, (524.29 +/- 163.59) U/L and (764.63 +/- 246.79) U/L for the IR group, and (150.90 +/- 27.05) U/L and (298.15 +/- 47.68) U/L for the control group (standard error of the mean). CONCLUSION: A significant decrease in ALT and AST levels was observed in the L and IP groups when compared to the ILR and IR groups (P < 0.05), but no significant difference in ALT and AST levels was observed in the L group when compared to the IP group (P > 0.05). These results suggest that pretreatment with lidocaine injected into the hepatoduodenal ligament prior to IR provides effective protection against subsequent IR injury to the liver. The novel approach of blocking innervation with lidocaine mimics hepatic IP, but is more convenient than IP at the time of liver surgery.