OBJECTIVE To explore the effect of respiratory syncytial virus(RSV)infection on the expression of key factors in the epithelial barrier of the human nasal epithelial cells(hNECs)derived from patients with chronic rhinosinusitis with nasal polyps(CRSwNP)and normal control mucosa.METHODS RSV with different multiplicity of infection(MOI)(0.1 and 0.3)infected hNECs derived from patients with CRSwNP(n=21)and normal control mucosa(n=9)for 24 h and 48 h,respectively.To detect the gene expression ZO-1,ZO-2,Claudin-1,Claudin-4,Occludin,E-cadherin and N-cadherin,total RNA was extracted and reverse transcribed into cDNA for real-time fluorescence quantification PCR.RESULTS The relative expression level of ZO-1,ZO-2,Claudin-1,Claudin-4,Occludin,E-cadherin and N-cadherin were decreased in hNECs post RSV infection.However,there was a statistical difference only in hNECs derived from CRSwNP(P<0.05).There was no significant difference in hNECs infected with RSV between eosinophilic CRSwNP(ECRSwNP)and non-eosinophilic CRSwNP(nonECRSwNP).CONCLUSION RSV infection could disrupt the epithelial barrier of the nasal mucosa,and patients with CRSwNP are more severely affected by RSV infection compared to healthy controls.The impact of RSV infection on mucosa between ECRSwNP group and nonECRSwNP group was no significant difference.

CRISPR-Associated Protein 9 ; Gene Editing ; CRISPR-Cas Systems/genetics*

Objective:To discuss the clinical efficacy of Xuesusheng granule for cancer related fatigue （CRF） deficiency of both Qi and blood in clinical practice， and to investigate its effect on immune function and quality of life. Method:According to random number table method， 118 patients were divided into observation group （59 cases） and control group （59 cases）. Patients in control group got Ejiao Huangqi oral liquid， 10-20 mL/time， 2 times/day. Patients in observation group got Xuesusheng granule， 10 g/time， 3 times/day. The treatment was continued for 8 weeks in both groups. Before and after treatment， scores of cancer fatigue scale （CFS）， functional assessment of cancer therapy （FACT） and Qi and blood deficiency syndrome were graded， and degree of fatigue was discussed with brief fatigue inventory scale-Chinese （BFI-C）. Levels of white blood cell count （WBC）， neutrophil count （NEU）， platelet count （PLT）， red blood cell count （RBC）， and hemoglobin （Hb） were compared before and after treatment.T lymphocyte subsets of （CD3⁺， CD4⁺， CD8⁺） levels， natural killer （NK） cells， interleukin-1（IL-1）， IL-6， IL-8 and tumor necrosis factor -α （TNF -α） were also detected， and the safety was evaluated. Result:After treatment the scores of each factor in CFS scale and the total score of CFS in the observation group were lower than those in the control group （P<0.01）， and the degree of fatigue was lower than that in control group （Z=2.101， P<0.05）. The total score of FACT and the scores of four dimensions of physiology， society/family， emotion and function in the observation group were lower than those in the control group （P<0.01）. Levels of WBC， NEU， RBC and Hb in the observation group were higher than those in the control group （P<0.01）. Scores of BFI-C， deficiency of Qi and blood were lower than those in control group （P<0.01）. Levels of NK， CD3⁺， CD4⁺ and CD4⁺/CD8⁺ in the observation group were all higher than those in the control group （P<0.01）， while the IL-1， CD8⁺， IL-6， IL-8 and TNF-α levels in observation group were lower than those in the control group （P<0.01）. Not observed， there was no adverse reaction related to Xususheng granule. Conclusion:Xuesusheng granule can improve the symptoms of fatigue， reduce the degree of fatigue， improve the quality of life， reduce the degree of bone marrow suppression， improve immune function， and regulate immune inflammatory factors with high clinical efficacy in the use for patients with CRF and deficiency of Qi and blood.

Objective:To explore the correlation between serum high mobility group protein B1 (HMGB1) and severity of coronary artery disease.Methods:From January 2018 to March 2019, 170 patients who underwent coronary angiography and were definitely diagnosed with coronary heart disease in Beijing Anzhen Hospital were divided into single-vessel lesion group(65 cases), double-vessel lesion group(55 cases) and three-vessel lesion group(50 cases) according to the results of coronary angiography.Sixty healthy persons in the same period were set as the control group.The serum levels of HMGB1, endothelin-1, C-reaction protein (CRP), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) were detected, and the severity of coronary artery disease was evaluated according to Gensini score standard.Results:The serum HMGB1 of patients with single vessel disease was (7.35±0.75) μg/L, that of double vessel disease group was (11.56±1.16) μg/L, that of three vessel disease group was (14.36±1.46) μg/L, and that of control group was (3.22±1.52) μg/L.The difference between the two groups was statistically significant ( F=14.235, P<0.001). The serum endothelin-1 in single vessel disease group was (198.56±19.86) ng/L, that in double vessel disease group was (226.54±22.64) ng / L, that in three vessel disease group was (435.65±43.95) ng/L, and that in control group was (120.47±13.27) ng/L.The difference between the two groups was statistically significant ( F=16.337, P<0.001). The serum CRP of patients with single vessel disease was (6.21±0.61) ng/L, that of double vessel disease group was (8.54±0.84) ng/L, that of three vessel disease group was (11.36±1.16) ng/L, and that of control group was (3.39±1.56) ng/L.The difference between the two groups was statistically significant ( F=15.206, P<0.001). Serum LDL-C was (3.23±0.33) mmol/L in single vessel disease group, 4.12±0.42 mmol/L in double vessel disease group, (6.23±0.63) mmol/L in three vessel disease group and (2.25±1.45) mmol/L in control group.The difference between the two groups was statistically significant ( F=22.017, P<0.001). Serum HDL-C was (4.02±0.42) mmol/L in single vessel disease group, (2.35±0.25) mmol/L in double vessel disease group, (1.79±0.29) mmol/L in three vessel disease group and (4.60±1.69) mmol/L in control group.The difference between the two groups was statistically significant ( F=18.564, P<0.001). The Gensini score of single vessel disease group was (10.36±2.26), that of double vessel disease group was (16.74±1.04) and that of three vessel disease group was (23.36±2.36). The difference between the two groups was statistically significant ( F=23.014, P<0.001). The levels of HMGB1, endothelin-1, CRP and LDL-C in patients with coronary heart disease were significantly higher than those in the control group, while the levels of HDL-C were significantly lower than those in the control group (all P<0.05). The higher the number of diseased vessels, the higher the levels of HMGB1, endothelin-1, CRP, LDL-C and Gensini score, the lower the level of HDL-C.Multiple linear regression analysis showed that HMGB1, endothelin-1, CRP and LDL-C were risk factors of Gensini score (standard coefficient was 0.480, 0.087, 0.173, 0.197, t=8.351, 9.047, 12.476, 11.692, all P<0.01); HDL-C level was a protective factor of Gensini score (standard coefficient -0.352, t value was 16.582, P<0.001). Pearson linear correlation analysis showed that serum HMGB1 level was positively correlated with endothelin-1, CRP and LDL-C ( r=0.536, 0.659, 0.724, all P<0.05), and negatively correlated with HDL-C ( r=-0.669, P<0.05). Conclusion:There are obvious inflammatory reaction and lipid metabolism disorder in patients with coronary heart disease.The level of HMGB1 in peripheral blood is closely related to the severity of coronary artery disease.

Objective: To observe the cardiomyocytes regeneration after myocardial infarction in Chinese small pigs, analyze the mechanism of myocardial fibrosis after myocardial regeneration. Methods: Nine Chinese small pigs, weight 18-20 kg, 6 pigs in the experimental group(to ligate LAD at the equal blood flow point), and 3 in the control group(no any operation was performed). Monitor the hemodynamics of the left ventricle. Cardiac specimens were taken after 4 weeks of LAD ligation, the left ventricle was divided into 17 segments, and fixed in 4% paraformaldehyde for 1 week. Hematoxylin-Eosin/(HE) staining, PTAH、Ki-67-DAB and α-sarcomeric-actin-DAB staining for pathological observation. Results: Four weeks of LAD ligation, different range of infarct size could be found in all the 17 segment of left ventricle. There was significant systolic pressure difference between the proximal and distal part in the left ventricular cavity. After 4 weeks of LAD ligation, there were a large number of new cardiomyocytes around the infarction area, which connected with the original mature cardiomyocytes directly. Large number of disassembled cardiomyocytes in the infarcted area and myocardial fibers were broken, cell structure disappeared, and nuclei were scattered in fibrotic tissues. Conclusion New cardiomyocytes after myocardial infarction were derived from the mature cardiomyocytes. Myocardial regeneration, cell disintegration, and myocardial fibrosis were performed synchronously under the influence of myocardial tension.

Objective To explore the efficacy of temporary titanium plate screwing for positional maintenance in reduction and internal fixation for displaced acetabular fractures.Methods A retrospective study was conducted of the 28 patients (28 hips) with displaced acetabular fracture who had been treated by open reduction and internal fixation from October 2013 to March 2016.They were 20 males and 8 females,aged from 24 to 68 years (average,42.3 years).The time from injury to surgery ranged from 7 to 21 days(average,14.5 days).According to the Letournel-Judet classification,there were 2 posterior column fractures,4 transverse fractures,5 posterior column plus posterior wall fractures,6 transverse plus posterior wall fractures,2 T-shaped fractures,3 anterior and posterior transverse fractures and 6 double column fractures.The posterior acetabular approach or combined anterior and posterior approach was used.In all the patients temporary titanium plate screwing was conducted to fix one side of the fracture so as to facilitate accomplishment of open reduction and internal fixation without losing the indirect anatomic reduction of the intraarticular fracture.After the open reduction and internal fixation was accomplished,the temporary screw fixation was removed in 26 patients but retained in 2 patients as needed.The reduction quality,complications and outcomes at the final follow-ups were recorded.Results All the 28 patients were followed up for 10 to 36 months (mean,15.6 months).By the Matta criteria,anatomical reduction was achieved in 26 cases,unsatisfactory reduction in one and poor reduction in one,yielding an anatomic reduction rate of 92.9％.By the improved Mere d'Aubigne & Postel criteria,the clinical outcomes at the final follow-up were excellent in 27 cases and good in one,yielding a good to excellent rate of 100％.Postoperatively,heterotopic ossification of different severities occurred in 11 cases but did not affect their joint function;transient paralysis of the sciatic nerve was reported in 6 cases but recovered 3 months after surgery.No complications like avascular necrosis of the femoral head or walking pain were observed during follow-ups.Conclusion Temporary fixation with titanium plate screws during open reduction and internal fixation for displaced acetabular fractures can effectively improve the reduction and fixation of the articular surface,leading to satisfactory short-term clinical outcomes.

Objective To evaluate the mean blood flow(MF) and pulsatility index(PI) of individual and sequential sa-phenous vein grafts in posterior descending artery(PDA) in the same patient undergone off-pump coronary artery bypass grafting (OPCABG).Methods 140 patients with coronary artery disease were studied.Sequential surgical technique using single large saphenous vein was used in grafts of diagnostic artery(DIAG)(side-side), obtuse marginal artery(OM)(side-side), and PDA( end-side) .Bulldog clamps were used to temporarily arrest the anastomoses of DIAG and OM near the large saphenous vein .Thus the sequential graft became to the single graft , the MF and PI of PDA were recorded in the simulating single graft by the transit-time flow measurement(TTFM), after that, bulldog clamps were removed and the MF and PI of PDA were recorded again in the real sequential graft.The data were compared.Six months later after surgery, coronary CTA were done in all pa-tients in clinic.Results The MF of single graft and sequential graft was(22.5 ±13.1)ml/min and(22.2 ±12.9)ml/min(P>0.05).The PI of single graft and sequential graft was 2.43 ±0.94 and 2.38 ±0.88(P>0.05).The patency of all grafts is good in all patients after 6 months follows-up.Conclusion There ars no hemodynamic differences between individual and sequential grafts in PDA.It is reasonable that the short-term surgical result is the same in individual and sequential grafts in PDA.

Objective To observe the efficacy of bloodletting therapy plus narrow band ultraviolet B (NB-UVB) in treating prurigo nodularis.Method According to the randomized controlled principle, the enrolled patients were divided into a treatment group and a control group. The treatment group was intervened by bloodletting cupping at the selected acupoints and the topical areas plus NB-UVB once every other day; the control group was by orally taking Mizolastine sustained release tablets and external application of Halometasone cream.Result The total effective rate was 85.7% in the treatment group versus 61.9% in the control group, and the difference was statistically significant (P<0.01).Conclusion Bloodletting therapy plus NB-UVB can produce a content efficacy in treating prurigo nodularis, with few adverse reactions.

BACKGROUND:The researches about the effect of retinoic acid on the proliferation of adipose-derived stem cells are rare, and the researches on the testosterone are mainly on the inhibition of cellaging. OBJECTIVE: To study the effects of retinoic acid and testosterone or combination on the cellcycle of adipose derived stem cells. METHODS:Adipose derived stem cells were isolated from adult female Sprague Dawley rats with 2 months age and cultured in vitro til passage 3 adipose derived stem cells, and then the 3rd passage adipose-derived stem cells were performed with adipogenic induction, osteogenic induction and surface marker identification. The cells were divided into six groups:(1) Control group;(2) 10-5 mol/L retinoic acid group;(3) Retinoic acid group;(4) 10-5 mol/L retinoic acid+testosterone group;(5) 10-6 mol/L retinoic acid+testosterone group;(6) Testosterone group. The adipose-derived stem cells in the control group were cultured with Dulbecco’s modified Eagle’s medium+10%fetal bovine serum culture medium, and the adipose-derived stem cells in the other five groups were induced with corresponding dose of retinoic acid and testosterone on the basis of control group. After cultured for 36 hours, the flow cytometry was used to detect the changes of cellcycle. RESULTS AND CONCLUSION:Compared with the control group, cellproportions in phase G 1 of 10-5 mol/L retinoic acid group and 10-6 mol/L retinoic acid group were increased significantly, and the cellproportions in phase S were decreased. Compared with control group, the cellproportion in phase G 1 of testosterone group was significantly reduced, and the cellproportion in phase S was increased. Compared with 10-5 mol/L retinoic acid group and 10-6 mol/L retinoic acid group, cellproportions in phase G 1 of 10-5 mol/L retinoic acid+testosterone group and 10-6 mol/L retinoic acid+testosterone group were reduced significantly and the cellproportions in phase S were increased. Retinoic acid can inhibit the cellcycle of adipose-derived stem cells in phase G 1 , and delay the process of the cellcycle from phase G1 to phase S;while testosterone can promote the cellcycle of adipose-derived stem cells from phase G1 to phase S;the combination induction of retinoic acid and testosterone can accelerate the process of the cellcycle of adipose-derived stem cells from phase G 1 to phase S.

OBJECTIVETo investigate whether adipose-derived stem cells (ADSCs) induced by retinoic acid (RA) in vitro express primordial germ cell marker alkaline phosphatase (ALP) and vasa.

METHODSADSCs were isolated from adult female SD rats and cultured in vitro. The third passage of ADSCs was identified by adipogenic differentiation, osteogenic differentiation and cell surface marker detection. The ADSCs were treated with 1×10(-5), 1×10(-6), or 1×10(-7) mol/L RA for 7 or 14 days, and the cellular expression of ALP was detected. vasa mRNA expression in ADSCs treated with 1×10(-5) mol/L RA for 7 days was detected using RT-PCR.

RESULTSThe OD value of ADSCs treated with 1×10(-5), 1×10(-6), or 1×10(-7) mol/L RA was 0.59∓0.04, 0.27∓0.07, and 0.15∓0.03 after a 7-day treatment, and was 0.42∓0.02, 0.34∓0.01, and 0.19∓0.02 after a 14-day treatment, respectively, demonstrating significantly enhanced ALP expression in RA-treated ADSCs compared with that in the control cells (0.07∓0.01 and 0.07∓0.01 at 7 and 14 days, respectively, P<0.01). The ADSCs showed a negative vasa mRNA expression after 1×10(-5) mol/L RA treatment for 7 days.

CONCLUSIONRA-induced ADSCs express ALP, a marker of primordial germ cells, but does not express the primordial germ cell marker vasa.

Adipose Tissue ; cytology ; Adult Stem Cells ; cytology ; enzymology ; Alkaline Phosphatase ; metabolism ; Animals ; Cell Differentiation ; Cells, Cultured ; Female ; Germ Cells ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tretinoin ; pharmacology