Objective To investigate the correlation of sarcoplasmic/endoplasmic reticulum Ca2+ATPase 2a(SERCA2a)and blood urea nitrogen(BUN)to albumin(ALB)ratio(BAR)with cardiac function and prognosis in patients with chronic heart failure(CHF).Methods A total of 200 CHF patients admitted to this hospital from January 2021 to January 2023 were selected as the CHF group and 100 healthy people un-dergoing physical examination as the control group.CHF patients were divided into class Ⅰ group(38 cases),class Ⅱ group(40 cases),class Ⅲ group(54 cases)and class Ⅳ group(68 cases)according to the New York Heart Association(NYHA)cardiac function classification after admission.According to the prognosis,they were divided into poor prognosis group(60 cases)and good prognosis group(140 cases).Serum SERCA2a BUN,ALB levels were measured and BAR was calculated.Spearman rank correlation coefficient was used to analyze the correlation between serum SERCA2a,BAR and NYHA cardiac function classification in CHF pa-tients.Taking the prognosis of CHF patients as the dependent variable,a multivariate unconditional Logistic regression model was established to determine its influencing factors.Receiver operating characteristic curve was drawn to evaluate the predictive value of serum SERCA2a and BAR for poor prognosis of CHF patients.Results Compared with the control group,the serum SERCA2a level was decreased and BAR was increased in the CHF group(P＜0.05).The level of SERCA2a in class Ⅰ group,class Ⅱ group,class Ⅲ group and classⅣ group decreased and BAR increased successively(P＜0.05).The NYHA classification was negatively cor-related with serum SERCA2a level(rs=-0.756,P＜0.05),and positively correlated with BAR(rs=0.722,P＜0.05).After 1-year follow-up,the incidence of poor prognosis in 200 CHF patients was 30.00％(60/200).Increased NYHA class(OR=1.936,95％CI:1.115-3.364),atrial fibrillation(OR=3.269,95％CI:1.078-9.913),N-terminal pro-B-type natriuretic peptide(OR=1.002,95％CI:1.001-1.009)and BAR(OR=1.169,95％CI:1.082-1.263)were independent risk factors for poor prognosis in patients with CHF(P＜0.05),increased left ventricular ejection fraction(OR=0.810,95％CI:0.716-0.916)and SERCA2a(OR=0.964,95％CI:0.947-0.981)were independent protective factors(P＜0.05).The area under the curve of serum SERCA2a combined with BAR to predict the poor prognosis of CHF patients was 0.865(95％CI:0.810-0.910),which was larger than 0.784(95％CI:0.720-0.839)and 0.777(95％CI:0.713-0.833)predicted by serum SERCA2a and BAR levels alone,and the difference was statistically significant(Z=2.944,3.250,P＜0.05).Conclusion The decrease of SERCA2a and the increase of BAR are closely re-lated to cardiac function and prognosis in patients with CHF.Serum SERCA2a combined with BAR has a higher value in predicting poor prognosis in patients with CHF.

Hypoxia-inducible factor (HIF-1α), a core transcription factor responding to changes in cellular oxygen levels, is closely associated with a wide range of physiological and pathological conditions. However, its differential impacts on vascular cell types and molecular programs modulating human vascular homeostasis and regeneration remain largely elusive. Here, we applied CRISPR/Cas9-mediated gene editing of human embryonic stem cells and directed differentiation to generate HIF-1α-deficient human vascular cells including vascular endothelial cells, vascular smooth muscle cells, and mesenchymal stem cells (MSCs), as a platform for discovering cell type-specific hypoxia-induced response mechanisms. Through comparative molecular profiling across cell types under normoxic and hypoxic conditions, we provide insight into the indispensable role of HIF-1α in the promotion of ischemic vascular regeneration. We found human MSCs to be the vascular cell type most susceptible to HIF-1α deficiency, and that transcriptional inactivation of ANKZF1, an effector of HIF-1α, impaired pro-angiogenic processes. Altogether, our findings deepen the understanding of HIF-1α in human angiogenesis and support further explorations of novel therapeutic strategies of vascular regeneration against ischemic damage.
Humans ; Vascular Endothelial Growth Factor A/metabolism* ; Endothelial Cells/metabolism* ; Transcription Factors/metabolism* ; Gene Expression Regulation ; Hypoxia/metabolism* ; Cell Hypoxia/physiology*

Objective:To investigate the effect of a degradable high-purity magnesium screw in fixing the greater trochanter bone flap of a lateral circumflex femoral artery transverse branch in the treatment of ischemic necrosis of femoral head in young and middle-aged adults.Methods:From February 2017 to February 2019, 12 cases (15 hips) of young and middle-aged patients with avascular necrosis of femoral head were treated in the Department of Orthopaedic of Affiliated Zhongshan Hospital of Dalian University. The age of patients was 30-53 years old. According to Association Research Circulation Osseous (ARCO), 2 hips were graded in stage II b, 4 in ARCO II c, 1 in ARCO III a, 5 in ARCO III b, 2 in ARCO III c and 1 in ARCO IV. The greater trochanter bone flap with a lateral circumferential vascular branch was used to fill the necrotic area, and fixed by a biodegradable high purity magnesium screw in the bone flap transfer. At 3, 6 and 12 months postoperation, the patient came to the hospital outpatient clinic for follow-up, and then were reviewed once a year. Imaging efficacy was evaluated by comparing preoperative and postoperative imaging. The Harris score and Visual Anoalogue Scale (VAS) score were tested at 12 and 24 months after surgery. The Harris score and VAS score before and after surgery were compared by Friedman test, and P<0.05 was considered statistically significant. Results:All 12 patients (15 hips) were entered in the 24-36 months of follow-up. At 12 and 24 months after surgery, Harris score was found at 87 (86, 92) and 90 (87, 92) respertively, which were both higher than that before surgery [59 (52, 74)] with a significant statistical difference ( Z=-3.743, Z=-4.473, P<0.05). However, there was no significant difference in Harris scores between 12 and 24 months after the surgery ( Z=-0.730, P>0.05). At the 12 and 24 months after surgery, VAS score was found at 3 (2, 3) and 2 (1, 3) respertively, which were both lower than that before surgery [6 (5, 6) ] with a significant statistical difference ( Z=-3.560, Z=-4.656, P<0.05). There was no statistical difference in VAS scores between 12 and 24 months after surgery ( Z=-1.095, P>0.05). X-ray and CT scan showed that the bone flaps healed well and the areas of osteonecrosis were repaired. Thirteen femoral heads were in good shape, and 2 femoral heads had further collapse of hips. No patients underwent joint replacement surgery at the time of last follow-up. Conclusion:Fixation of the greater trochanter flap of lateral circumflex femoral artery transverse branch with a degradable high-purity magnesium screw can ensure the healing of the flap at the implantation site and avoid the displacement and shedding of the flap. It is a new therapeutic option to treat the avascular necrosis of femoral head of young and middle-aged people.

Inguinal incarcerated hernia in adults is a common acute abdomen in hernia and abdominal wall surgery. If not treated in time, it is easy to progress to constrictive hernia, lead to intestinal ischemic necrosis, cellulitis of tegmental tissue outside the hernia, hernia sac empyema, intestinal fistula, and even cause toxic shock, with significantly increased mortality. The types of incarceration are different and the corresponding management methods are different. Based on the proposal of the concept of musculopubic foramen hernia repair, inguinal incarcerated hernia includes incarcerated indirect hernia, incarcerated direct hernia, incarcerated femoral hernia, etc. At present, there is no uniform standard for the comprehensive treatment of different types of inguinal incarcerated hernia, and the clinical management strategies of adult inguinal incarcerated hernia still face serious challenges.

Objective:To dynamically trace the migration and therapeutic effects of human bone marrow mesenchymal stem cells (MSCs) in mice with liver injury after cell transplantation through in vivo bioluminescent imaging (BLI).Methods:The MSCs were transfected with the lentivirus CMV-Luciferase2-mKate2 and mKate2 positive cells were purified and screened by fluorescence-activated cell sorting (FACS) after 96 h. The purified MSCs-R (MSCs-CMV-Luciferase2-mKate2) were used by in vitro and in vivo BLI. The mice (male BALB/c nude mice) were divided into 4 groups with 9 mice per group by random number table method, including (1) Liver injury experimental group: The liver injury model was established by intraperitoneal injection of CCl 4, and MSCS-R transplantation through spleen injection was performed 24 h later; (2) Control experimental group: The same volume of phosphate buffer (PBS) was injected intraperitoneally, and MSCS-R transplantation through spleen injection was performed 24 h later; (3) Liver injury group: Liver injury model was established and PBS was injected into the spleen;(4) Blank group: The mice were intraperitoneally injected of PBS.BLI was performed daily after cell transplantation until light signals disappeared in the liver region, and the pathological examination of liver tissue was obtained 14 d after MSCs-R transplantation. Linear regression analyses were performed to determine the correlation between the optical signal intensity and the number of cells, and statistical differences of the optical signal intensity between liver injury experimental group and control experimental group were evaluated using the Student′s t test. Results:The MSCs were readily transfected with lentivirus CMV-Luciferase2-mKate2 for 96 h. The transfected MSCs were purified by FACS and more than 95% of MSCs were mKate2 positive. The optical signal intensity of MSCs-R detected by BLI in vitro significantly correlated with cell numbers in vitro (R 2=0.980). In both of liver injury experimental group and control experimental group, cell migration to the liver was observed on the first day after intrasplenic transplantation of MSCs-R, and the optical signal intensity in the area of liver of liver injury experimental group was higher than that of control experimental group ( t=15.476, P<0.001). The optical signal intensity in the hepatic area was observed in 11 d after transplantation in liver injury experimental group, compared to control experimental group in 5 d. Optical signal was not detected in mice in the other two groups. Histopathology showed that the degree of liver injury after MSCs-R transplantation was significantly lower in liver injury experimental group than control experimental group. Conclusions:The dynamical migration of MSCs transplanted to the spleen and settled in the damaged liver could be tracked by BLI, and liver injury can prompt MSCs directionally migrate to the damaged tissues and play their role in repairing liver injury.

Background and Objectives: Tracking of the tumor progression by MSCs-based therapy is being increasingly important in evaluating relative therapy effectively. Herein, Bioluminescence imaging (BLI) technology was used to dynamically and quantitatively track the hepatocellular carcinoma suppressive effects by human umbilical cord mesenchymal stem cells (UC-MSCs). Methods: and Results: The stem cells present typical phenotypic characteristics and differentiation ability by morphology and flow cytometry analysis of marker expression. Then, the growth inhibition effect of conditioned medium and UC-MSC on H7402 cells was studied. It is found both the conditioned medium and UC-MSC can effectively decrease the proliferation of H7402 cells compared with the control group. Meanwhile, the relative migration of UC-MSC to H7402 is also increased through the transwell migration assay. In addition, a mice hepatoma tumor model was built by H7402 cells which can express a pLenti-6.3/DEST-CMV-luciferase 2-mKate2 gene. The effect of stem cells on growth inhibition of tumor in a mice transplantation model was dynamically monitored by bioluminescence imaging within 5 weeks. It has shown the bioluminescence signal intensity of the tumor model was significantly higher than that of the UC-MSC co-acting tumor model, indicating that the inhibition of UC-MSC on liver cancer resulted in low expression of bioluminescent signals. Conclusions The microenvironment of UC-MSCs can effectively inhibit the growth of liver cancer cells, and this therapeutic effect can be dynamically and quantitatively monitored in vivo by BLI. This is of great significance for the imaging research and application of stem cells in anticancer therapy.

Objective To investigate the effects of melatonin on the proliferation of neural stem cells (NSCs) in cerebral ischemia reperfusion (IR) rats, and to explore the possible mechanisms. Methods Seventy-two rats were randomly divided into the normal control group (n=12), model group (n=30) and melatonin group (n=30) according to the random number table. The rats in the model group and melatonin group were divided into four subgroups: 6 h, 24 h, 72 h and 7 d subgroups according to the time after IR. The morphological changes of the subventricular zone (SVZ) were examined by HE staining;the effects of melatonin on NSCs proliferation were examined by immunofluorescence staining;the effects of melatonin on toll-like receptor 4 (TLR4) and nuclear factor (NF)-κB p65 protein were examined by immunohistochemistry staining and Western blotting analysis. The correlation between the proliferating NSCs and TLR4 protein or the NF-κB p65 protein was analyzed by linear regression analysis. Results HE staining showed that the cells in the SVZ of rats in the model group were in disorder and irregular in shape. In the melatonin group, the cells in the SVZ of the injured side were relatively well arranged. Immunofluorescence staining showed that the number of proliferating cell nuclear antigen (PCNA)+Nestin+4',6-diamidino-2-phenylindole (DAPI)+cells in the SVZ of the model (498.47 ± 26.44/mm2) and melatonin groups (623.10 ± 39.70/mm2) increased gradually, and reached a higher level after IR for 7 days, which were significantly higher than the normal control group (203.91 ± 32.23/mm2) (F=35.193, 170.344, 277.536, 285.947, all P＜0.01). The number of PCNA+Nestin+DAPI+cells in the melatonin group rats at each time points was significantly higher than that in the model group (F=102.561, 91.244, 168.502, 38.013, all P＜0.01). Immunohistochemistry staining showed that the numbers of TLR4+and NF-κB p65+cells in the SVZ of the model (740.02±31.63/mm2;710.01± 26.59/mm2) and melatonin groups (555.57 ± 25.28/mm2;528.85 ± 30.60/mm2) increased gradually, and reached a higher level 7 d after IR, which were significantly higher than the normal control group (107.97±12.84/mm2;109.80±13.89/mm2) (F=21.413, 263.059, 873.691, 1 037.098, all P＜0.01;F=26.374, 372.940, 854.826, 929.018, all P＜0.01). There were less TLR4+(F=7.641, 25.135, 66.094, 103.753, all P＜0.05) and NF-κB p65+cells (F=18.612, 69.597, 113.113, 119.814, all P＜0.01) in the melatonin group as compared with those in the model group at each time points. Western blotting analysis showed that the expression of TLR4 (0.87±0.08;0.68±0.06) and NF-κB p65 (0.72±0.05;0.58±0.05) protein was higher in the model and melatonin groups as compared with the normal control group (0.35±0.04, 0.31±0.03;F=107.43, F=132.51, both P＜0.01). The expression of the TLR4 and NF-κB p65 protein was lower in the melatonin group as compared with that in the model group (P＜0.01). Linear regression analysis showed that the differences of PCNA+Nestin+DAPI+cells were all negatively correlated with that of the TLR4+cells and NF-κB p65+cells in the melatonin group (r2=0.838, r2=0.813, both P＜0.01). Conclusion Melatonin can inhibit the expression of TLR4 and NF-κB p65 protein, thus promote the proliferation of endogenous NSCs in cerebral ischemia reperfusion rats.

Objective To summarize the experience of surgical treatment of severe ventricular septal defect (VSD) accompanied by pulmonary hypertension (PH) in little infants. Methods The clinical data of 11 patients with VSD accompanied by PH admitted to the Department of Cardiac Surgery of Tianjin Children's Hospital of the Pediatric Clinical College in Tianjin Medical University from January 2016 to January 2017 were retrospectively analyzed. There were 6 males and 5 females, with the ages of 3 - 6 months, mean (5.36±1.03) months and body weights 4.2 - 7.5 kg, mean (5.9±1.0) kg. Preoperatively, all patients had recurrent pneumonia and heart failure history. After 2 - 3 times of medical treatment in hospitals, the patients underwent sub-acute radical operation under cardiopulmonary bypass. All patients were followed-up with echocardiography, chest X-ray and electrocardiogram examinations at 3, 6 and 12 months after heart surgery. Results After operation, all the children spontaneously recovered the heart beats, the stay times in surgical intensive care unit (SICU) were 2 - 7 days, and total hospitalization times were 12 - 17 days. All patients were followed up for 12 months, no death occurred in the whole group, and the heart and lung functions recovered satisfactorily. Conclusions Little infants with large VSD and PH should undertake operation as early as possible. The patients with recurrent pneumonia, heart failure in a short term should receive medical and surgical doctors' cooperative treatment, and the disease situation ought to be adjusted with one's best ability to a stable status when the sub-acute surgery can be carried out safely.

BACKGROUND:Now experimental and clinical research on suitable bone substitutes for alveolar bone defects after dental implantation is an issue of concern.OBJECTIVE:To study the therapeutic effect of titanium core/bone morphogenetic protein (BMP) composite material on alveolar bone defects after immediate implant placement.METHODS:Twenty-four New Zealand rabbits were randomly assigned into normal group (no intervention),experimental group or control group.Animal models of bone femoral greater trochanter defect were made in the experimental and control groups.Dental implant and titanium core/BMP composite material were implanted in the experimental group,while dental implant and titanium core were implanted in the control group.Percentage of CD4+,CD8+ T lymphocytes,natural killer cell activity and interleukin 2 level were detected at postoperative 4 weeks;bone mineral density and osteogenesis around the implant were detected at postoperative 16 weeks through X-ray and histological examinations.RESULTS AND CONCLUSION:X-ray results showed that the bone mineral density in the experimental group was significantly higher than that in the control group (P ＜ 0.05).Histological results showed that in the experimental group,different degrees of cell lysis around the composite,more bone cells and bone matrices were found,implant-bone osseointegration formed well,and red-dyed mature bone tissues were detective inside the implant.Compared with the experimental group,lower number of bone cells and fibrocytes were found in the control group.Additionally,the percentage of CD4+ and CD8+ T lymphocytes,natural killer cell activity and interleukin 2 level in the experimental group were significantly lower than those in the control group (P ＜ 0.05).To conclude,the titanium core/BMP composite material can effectively repair alveolar bone defects after immediate implant placement to guide the growth of bone cells.

BACKGROUND:Nerve growth factor has been shown to play an important role in bone healing, but little is reported on the effect of growth factor composite scaffolds via the immediate implantation in the repair of canine bone defects. OBJECTIVE:To analyze the effect of nerve growth factor composite scaffolds via the immediate implantation for the repair of canine bone defects. METHODS:Nerve growth factor composited strontium apatite scaffolds were prepared. Canine mandibular defect models were established and divided into three groups, fol owed by implanted with composite scaffold (experimental group), strontium apatite (positive control group), or nothing (blank control group). The three-dimensional CT reconstruction and hematoxylin-eosin staining of canine mandibular bone defects were observed. RESULTS AND CONCLUSION:In the blank control group, there were few new bones surrounding bone defect. Trabecular bones spread from the defect center to the surrounding tissues in the experimental and positive control groups. The bone density, volume, thickness, and implant-bone contact were significantly increased, while the trabecular separation was significantly decreased in the experimental group compared with the positive control and blank control groups (P<0.05), and al above indicators in the positive contro group were significantly higher than those in the blank control group (P<0.05). Hematoxylin-eosin staining showed that in the experimental group, there were a large number of new bones that contacted with the surrounding bones closely, and trabecular bones arranged regularly. In the positive control group, newborn osteoid, trabeculare, and a smal amount of debris were found. In the blank control group, few new bones were connected with the surrounding bones untightly and trabecular bone arranged irregularly. These results indicate that the nerve growth factor composite scaffold can promote the bone regeneration in the canine bone defects after immediate implantation.