OBJECTIVES: To investigate the mechanism of DDX17 for regulating proliferation and migration of pulmonary arterial smooth muscle cells (PASMCs) during the development of pulmonary hypertension (PH). METHODS: In murine PASMCs cultured under normoxic or hypoxic conditions, the effects of transfection with si-Ddx17 and insulin treatment, alone or in combination, on cell proliferation and migration were evaluated using Ki-67 immunofluorescence staining, scratch assay and Transwell assay. Western Blotting was performed to detect the changes in protein expression levels of DDX17, 4EBP1, S6, p-4EBP1, and p-S6. In a mouse model of PH induced by intraperitoneal injection of monocrotaline (MCT), the changes in pulmonary vasculature were examined using HE staining following tail vein injection of AD-Ddx17i. RESULTS: The PASMCs in hypoxic culture exhibited significantly enhanced cell proliferation and migration and protein expressions of p-4EBP1 and p-S6, and these changes were obviously reversed by transfection with si-Ddx17. Treatment with insulin significantly attenuated the effect of si-Ddx17 against hypoxic exposure-induced changes in PASMCs. In the mouse model of MCT-induced PH, transfection with AD-Ddx17i obviously alleviated pulmonary vascular stenosis and intimal hyperplasia. CONCLUSIONS The expression of DDX17 is elevated in hypoxia-induced PASMCs and PH mice, and silencing DDX17 significantly inhibits PASMC proliferation and migration in vitro and pulmonary vascular remodeling in PH mice by reducing mTORC1 activity.
Animals ; Cell Proliferation ; Cell Movement ; DEAD-box RNA Helicases/metabolism* ; Myocytes, Smooth Muscle/metabolism* ; Mice ; Pulmonary Artery/cytology* ; Hypertension, Pulmonary/metabolism* ; Mechanistic Target of Rapamycin Complex 1 ; Cells, Cultured ; Muscle, Smooth, Vascular/cytology*

Objective:To investigate the effect of L-carnitine on myocardial ischemia-reperfusion injury in patients undergoing coronary artery bypass grafting (CABG) operation.Methods:The clinical data of 60 patients who had underwent CABG under extracorporeal circulation in Department of Cardiovascular Surgery, the First Affiliated Hospital of Medical College of Xi′an Jiaotong University from January 2015 to December 2017 was retrospectively analyzed. L-carnitine was infused into the patients in the observation group while the equal amount of normal saline was given to the patients in the control group during perioperative period. Venous blood was collected from each patient 1 hour before the surgery and at 2, 6, 24 and 72 hours after unclamping the aorta, then serum levels of aspartate aminotransferase (AST), lactic acid dehydrogenase (LDH), creatine kinase (CK), and creatine kinase-MB isozyme (CK-MB) were detected. Additionally, cardiac function indices including stroke volume (SV), left ventricular diastolic diameter (LVDD), cardiac output (CO), Left ventricular ejection fraction (lVEF) were compared between the two group patients before the surgery and 6 days after operation.Results:Compared with before the operation, the levels of serum enzymes were all significantly increased after unclamping the aorta. The levels of serum AST, LDH and CK reached their peak at 24 hours after unclamping the aorta. Notably, the levels of serum AST, LDH and CK in the experimental group were lower than that in the control group at each time point after unclamping the aorta ( P<0.05). For serum CK-MB, the level in the experimental group reached its peak at 6 hours after unclamping the aorta, while the peak value was observed at 24 hours after unclamping the aorta in the control group. The level of CK-MB in the experimental group was significantly decreased compared with the control group at each time point after unclamping the aorta ( P<0.05). Moreover, there were no significant differences in cardiac function indices between the 2 groups before the operation ( P>0.05). The levels of SV, CO and LVEF in the experimental group were significantly increased while the level of LVDD was obviously decreased compared to that in the control group after the operation ( P<0.05). Conclusions:L-carnitine has protective effects on myocardial ischemia-reperfusion injury in patients undergoing CABG operation under cardiopulmonary bypass, with reducing serum levels of myocardial enzyme in these patients.

Objective:To investigate the clinical effect of lateral rectus abdominis approach combined with presacral decompression for surgical treatment of old Denis type II sacral fractures complicated with upper sacral plexus injury.Methods:A retrospective case series study was performed on the clinical data of 9 patients with old Denis type II sacral fractures complicated with upper sacral plexus injury (L 4-S 1) admitted to Zhongnan Hospital of Wuhan University from June 2010 to December 2016. There were 6 males and 3 females, aged (33.1±7.5)years (range, 15-58 years). Embolization of internal iliac artery and preimplantation of abdominal aortic balloon were performed 2 hours before operation under the guidance of digital subtraction angiography (DSA). Surgery was performed using a single lateral rectus abdominis approach combined with presacral decompression. The operation time, intraoperative blood loss and full weight-bearing time were recorded. The visual analogue scale (VAS) and European QOL Five Dimensional health scale (EQ-5D) were compared before and after operation. The Gibbons' impairment scale was used to assess neurological function. X-ray and CT scan were used to observe internal fixation and fracture healing. The complications during and after operation were recorded. Results:The patients were followed up for 24-52 months [(35.2±5.2)months]. The operation time was (2.9±0.6)hours. The intraoperative bleeding was (573±138)ml, and the full weight-bearing time was (11.6±1.2)weeks. X-ray and CT scan showed bone healing in all patients at the latest follow-up. The VAS and EQ-5D scale improved from preoperative (7.8±0.6)points and (0.34±0.07)points to the final follow-up of (0.8±0.3)points and (0.81±0.05)points ( P<0.05). According to Gibbons classification, 8 patients were grade I and 1 patient was grade II one year after operation ( P<0.01). Namely, the radiation pain of lower extremities was significantly improved in all patients, among which 8 patients showed pain disappeared and completely returned to normal and 1 patient showed residual numbness and hypoesthesia of the affected limbs. No major complications (eg, iatrogenic lumbosacral plexus injury, vital blood vessels or pelvic organs injury) occurred during the operation. During the follow-up period, only one patient developed traumatic hip arthritis and underwent total hip arthroplasty 6 months after operation. Fractures of the remaining patients were healed well without complications like infection, pressure ulcer or implant failure. Conclusions:For old Denis type II sacral fractures complicated with upper sacral plexus injury, lateral rectus abdominis approach combined with presacral decompression can fully decompress the upper sacral plexus nerve, relieve pain, and promote functional rehabilitation, with low incidence of complications. It is an alternative surgical method for the treatment of old Denis type II sacral fractures complicated with upper sacral plexus injury.

Objective To investigate the effect of intravenous drip of levocarnitine during perioperative period on serum cytokines and cardiac troponin (cTn) I in patients undergoing coronary artery bypass grafting (CABG) under extracorporeal circulation. Methods The clinical data of 70 patients who had underwent CABG under extracorporeal circulation in Department of Cardiovascular Surgery, the First Affiliated Hospital of Medical College of Xi′an Jiaotong University from January 2015 to December 2017 were retrospectively analyzed. Among them, 35 cases were treated with intravenous infusion of levocarnitine (experiment group), and 35 cases were not treated with intravenous infusion of levocarnitine (control group). The serum levels of cTnI, tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-8 and IL-10 1 h before operation and 2, 6, 24, 72 h after aorta open were detected; the postoperative recovery and complication were recorded. Results There were no statistical differences in TNF-α, IL-6, IL-8, IL-10 and cTnI before operation between 2 groups (P＞0.05); the indexes 2, 6, 24 and 72 h after aorta open were significantly higher than those before operation, and there were statistical differences (P＜0.05). The TNF-α, IL-6 and cTnI 2, 6, 24 and 72 h after aorta open in experiment group were significantly lower than those in control group, and the IL-10 was significantly higher than that in control group; the IL-8 2, 6 and 24 h after aorta open in experiment group were significantly lower than that in control group, and there were statistical differences (P＜0.05). The ICU monitoring time, electrocardio-monitoring time, duration of antibiotic treatment, duration of drainage tube, mechanical ventilation time and length of hospital stay in experiment group were significantly shorter than those in control group: (2.9 ± 0.5) d vs. (3.5 ± 0.8) d, (5.7 ± 1.8) d vs. (7.6 ± 3.2) d, (6.7 ± 1.5) d vs. (9.8 ± 2.2) d, (3.1 ± 0.8) d vs. (3.9 ± 1.4) d, (3.3 ± 2.1) d vs. (5.1 ± 2.3) d and (8.1 ± 2.2) d vs. (12.8 ± 2.6) d, and there were statistical differences (P＜0.01). Moreover, there were no severe perioperative complications such as myocardial infarction and pulmonary infection in 2 groups. Conclusions Intravenous drip of levocarnitine during perioperative period could effectively control postoperative inflammatory response and myocardial injury in patients undergoing CABG under extracorporeal circulation. The effect of levocarnitine may be related to its ability to balance the levels between proinflammatory cytokines and anti-inflammatory cytokines.

OBJECTIVE: To obtain cancer stem cells (CSCs) from surgically resected colorectal cancer specimens and identify their stem cell characteristics. METHODS: Colorectal cancer tissue specimen obtained from a patient undergoing radical resection of colorectal cancer were implanted in nude mice, and the xenograft was harvested 1 month later to obtain purified tumor cells by enzyme digestion and adherent culture. The CSCs were screened by limiting dilution method and serum-free culture to identify their phenotypes. Soft agar colony assay was used to assess the proliferative ability of the CSCs and human colorectal cancer cell line SW480. The tumorigenic ability of the isolated CSCs and SW480 cells was evaluated by observing their subcutaneous tumor formation in nude mice. Western blotting and immunofluorescence assay were used to detect the immunophenotype of the CSCs and SW480 cells. RESULTS: The primary cultured CSCs from clinical specimens of colorectal cancer underwent differentiation in the presence of serum in the culture. Soft agar colony formation assay showed that the CSCs had a colony formation rate above 50%, significantly higher than the rate of colorectal cancer SW480 cells (4.41%; < 0.01). In nude mice, subcutaneous injection of 500 CSCs was sufficient to result in subcutaneous tumor formation, while the injection of 500 SW480 cells failed to form any subcutaneous tumors. The CSCs expressed CD133 and CD44 but not CK7, while SW480 cells expressed CK7 but not CD133 or CD44. CONCLUSIONS CSCs can be derived by primary culture of cancer cells obtained from surgically resected colorectal cancer tissue followed by serum-free culture, and the CSCs obtained have self-renewal and differentiation abilities.
Animals ; Cell Culture Techniques ; Cell Differentiation ; Cell Line, Tumor ; Colorectal Neoplasms ; Humans ; Mice ; Mice, Nude ; Neoplastic Stem Cells

We analyzed the clinicopathological data of 3 cases of primary intraosseous hematopoietic pseudotumor (IHPT), which had been previously misdiagnosed as malignancies or metastases both clinically and pathologically. Two of the patients received close follow-up for 132 and 100 months, and one patient was lost to follow-up, and the tumors were confirmed to be benign in all the 3 cases. IHPT is a rare benign intraosseous solid lesion consisting of tissues resembling normal hematopoietic tissue, and can be easily misdiagnosed as malignancy. Understanding the clinicopathological features and the outcomes of the disease can facilitate the clinical decisions on individualized diagnosis and therapeutic regimens.
Bone Marrow ; Follow-Up Studies ; Hematopoietic Stem Cell Transplantation ; Humans

Objective: To investigate the effect of Calcium dibutyryladenosine cyclophosphate on myocardial ischemia-reperfusion injury in patients undergoing coronary artery bypass grafting (CABG) operation. Methods: The clinical data of 62cases were retrospectively analyzed. These patients were performed coronary artery bypass graft (CABG) operation under cardiopulmonary bypass (CPB) in the First Affiliated Hospital of Medical College of Xi′an Jiaotong University from January 2013 to January 2016. According to the use calcium dibutyryladenosine cyclophosphate, 62 patients were divided into the control group and the experimental group. Thirty-one cases in the experimental group were given an intravenous drip of calcium dibutyryladenosine cyclophosphate, while other 31 cases in the control group were not given to calcium dibutyryladenosine cyclophosphate during perioperative period. Venous blood was collected from each patient 1 h before the surgery and 2, 6, 24 and 72 h after unclamping the aorta, then serum levels of aspartate aminotransferase (AST), lactic acid dehydrogenase (LDH), creatine kinase (CK), and creatine kinase-MB isozyme (CK-MB) were detected. Additionally, cardiac function indices including SV, LVDD, CO, LVEF in these patients were recorded before the operation and 6 days after operation. Results: Compared with those before the operation, the levels of serum enzymes were all significantly increased after unclamping the aorta. The levels of serum AST, LDH and CK reached their peak 24 h after unclamping the aorta. Notably, the levels of serum AST, LDH and CK in the experimental group were lower than those in the control group at each time point after unclamping the aorta (P < 0.05). For serum CK-MB, the level in both the two groups reached its peak 6 h after unclamping the aorta. The level of CK-MB in the experimental group was significantly decreased compared with that in the control group 6, 24 and 72 h after unclamping the aorta (P < 0.05). Moreover, there were no significant differences in cardiac function indices between the 2 groups before the operation (P > 0.05). The levels of SV, CO and LVEF in the experimental group were significantly increased while the level of LVDD was obviously decreased compared to that in the control group after the operation, SV: (70.93 ± 9.72) ml vs. (65.07 ± 6.81) ml, CO: (5.32 ± 0.98) L vs. (4.29 ± 0.91) L, LVEF: (57.87 ± 8.92)% vs. (52.68 ± 7.31)% and LVDD: (55.67 ± 4.12) mm vs. (62.85 ± 5.71) mm, and there were statistical differences (P < 0.05). Conclusions Calcium dibutyryladenosine cyclophosphate has protective effects on myocardial ischemia-reperfusion injury and cardiac function in patients undergoing CABG operation under CPB, by reducing serum levels of myocardial enzymes in these patients.

Objective To explore the antigen presentation of CT26.WT via intra-peritoneal injection. Methods The intra-peritoneal injection model was made via injecting cell suspensions in mice. The spleen was isolated from BALB/c mice toco-culture with CT26.WT to detect tumor-killed ability. Phenotype identification methods and CCK8 massy were used to measure the ability of antigen presentation and stimulate T lymphocyte proliferation. IHC was used to detect the expression of B7H4 in normal and tumor tissues. Results Along with the extension of intra-peritoneal injection, the surviving number of cells was increased, contrary to the apoptosis. DC cells failed in maturation and impaired in stimulating T lymphocyte proliferation. B7H4 was higher in tumor tissues. Conclusions With the extension of intra-peritoneal injection, the mature DC cells were scared in number, resulting in the impairement of antigen-presentation. Moreover, the higher B7H4 expression in tumor tissues led to the lack of second signals which may stimulate T cells. Consequently, the ability of T cells in killing tumor cells was decreased so that they escape immunosurveillance.

OBJECTIVEThis study is in an attempt to evaluate the diagnostic significance to predict the spermatogenesis of azoospermic men in examination of serum follicle-stimulating hormone (FSH) combination with serum inhibin B (INHB).

METHODSQuantitative examination of serum FSH and INHB was performed in 95 case of azoospermic men. According to their classifications of testicular biopsy with histopathological examination, there were 20 patients of Sertoli cell only, 25 of hypospermatogenesis, 18 of spermatogenic maturation arrest (complete or incomplete), and 32 of normal spermatogenesis. The association of serum FSH and INHB levels with histopathological classifications were analyzed by using statistical software.

RESULTSSerum FSH, INHB and INHB/FSH levels of Sertoli cell only differed with statistical significance from hypospermatogenesis, spermatogenic maturation arrest and normal spermatogenesis (P<0.05). FSH, in which there were no statistical significance among the latter three classifications (P>0.05). Serum FSH, INHB and INHB/FSH levels were no relationship with maturation arrest (P>0.05), but were negatively related to the other classifications (P<0.05). INHB level less than 28.55 pg/ml predicted Sertoli cell only in a sensitivity of 97% and a specificity of 85%.

CONCLUSIONSerum FSH and INHB levels is ineffective to distinguish the spermatogenic classifications from azoospermic men, but they are available to confirm the disease of Sertoli cell only. The other abnormalities of azoospermic men is also dependent on bioptic histopathology to confirm the subtypes.

Adolescent ; Adult ; Azoospermia ; blood ; diagnosis ; Follicle Stimulating Hormone ; blood ; Humans ; Infertility, Male ; blood ; diagnosis ; Inhibins ; blood ; Male ; Middle Aged ; Oligospermia ; Spermatogenesis ; Testis ; physiology ; Young Adult

OBJECTIVETo assess the value of combined evaluation of serum follicle-stimulating hormone (FSH), inhibin B (INHB), chromosome karyotyping and AZF microdeletion of Y-chromosome (AZF-MD-Ych) in predicting the success of testicular sperm aspiration (TESA) in azoospermic patients.

METHODSA total of 262 azoospermic patients were divided into two groups with normal (n=162) and abnormal (n=100) serum FSH levels. INHB levels, INHB/FSH ratio, chromosome karyotype patterns of the peripheral lymphocytes, and AZF-MD-Ych were compared between the two groups. Among the patients receiving TESA, the success rate of the procedure was compared between the two groups after excluding abnormalities in INHB, chromosome karyotype and AZF-MD-Ych.

RESULTSSignificant differences were found between the two groups in serum INHB level, INHB/FSH and chromosome karyotypes (P<0.05), but not in AZF-MD-Ych (P>0.05). After excluding the abnormalities in chromosome karyotypes, AZF-MD-Ych and INHB, sperms were obtained successfully by TESA from 61.82% (34/55) of patients with normal FSH but from none of those with abnormal FSH (P<0.01).

CONCLUSIONA combined evaluation of serum FSH, INHB, chromosome karyotypes and AZF-MD-Ych can effectively predict the success of TESA in azoospermic patients, and abnormalities in all the 4 indices suggest a very low success rate of sperm retrieval by TESA.

Azoospermia ; Chromosome Deletion ; Chromosomes, Human, Y ; Follicle Stimulating Hormone ; blood ; Humans ; Infertility, Male ; Inhibins ; blood ; Karyotyping ; Male ; Prognosis ; Sex Chromosome Aberrations ; Sex Chromosome Disorders of Sex Development ; Sperm Retrieval ; Spermatozoa ; Testis ; Treatment Outcome