BACKGROUND:Our previous studies have shown that mesenchymal stem cells play an immunomodulatory role in Th17 cells, but the mechanism of action remains to be elucidated, therefore, to explore the role of Tol-like receptor 1 in which wil provide possible experimental basis for the future potential of celltherapy strategies.
OBJECTIVE:To investigate the role of Tol-like receptor 1 in the immunoregulatory function of mesenchymal stem cells on Th17 cells.
METHODS:Separation of adherent bone marrow-derived human embryonic sources of mesenchymal stem cells, immunomagnetic separation of normal CD4+T cells. CD4+T cells were cultured alone or in combination with mesenchymal stem cells co-cultured 4d. Real-time quantitative polymerase chain reaction assay interleukin-17, Tol-like receptor 1 expression levels related genes;number of Th17 cells by flow cytometry. Bone marrow mesenchymal stem cells from human embryos were separated using adherence method, and co-cultured with human CD4+T cells from healthy donors using immunomagnetic separation method for 4 days. The expression of interleukin-17 and Tol-like receptor 1 mRNA was detected by real-time PCR, and the number of Th17 cells was observed by flow cytometry.
RESULTS AND CONCLUSION:Tol-like receptor 1 mRNA was expressed in both CD4+T cells and mesenchymal stem cells. Level of interleukin-17 mRNA was significantly higher in mesenchymal stem cells co-cultured with CD4+T cells than in CD4+T cells cultured alone (relative value 3.59±0.11 vs. 1.14±0.08, P<0.01). Consistent with the expression of interleukin-17 mRNA, increased level of Tol-like receptor 1 mRNA was detected in mesenchymal stem cells co-cultured with CD4+T cells compared with CD4+T cells cultured alone (relative value 6.07±1.79 vs. 1.53±0.63, P<0.01). Furthermore, Flow cytometry analysis showed that the percentage of Th17 cells in the mesenchymal stem cells co-cultured with CD4+T cells was significantly higher than that in CD4+T cells cultured alone, (4.53±1.27)%vs. (2.39±0.80)%(P<0.01). Tol-like receptor 1 might be involved in the immunoregulatory regulation of Th17 cells by mesenchymal stem cells.

BACKGROUND:Previous studies have found that embryonic bone marrow mesenchymal stem cel s can promote human Th17 cel proliferation, but the inherent regulatory mechanisms stil need to be elucidated. OBJECTIVE:To investigate the role of Tol-like receptor 3 in the immunoregulation of Th17 cel s by mesenchymal stem cel s.
METHODS:Human CD4+T cel s from healthy donors were isolated by immunomagnetic bead method, and then cultured alone or co-cultured with embryonic bone marrow mesenchymal stem cel s for 4 days. The mRNA expression level of interleukin-17, Tol-like receptor 3 and MyD88 was detected by real-time PCR.
RESULTS AND CONCLUSION:Compared with CD4+T cel cultured alone group, the mRNA level of interleukin-17 was significantly higher in the co-culture group (3.59±0.11 vs. 1.14± 0.08, P<0.01). Consistent with the expression of interleukin-17 mRNA, increased level of Tol-like receptor 3 mRNA was detected in the co-culture group compared with the CD4+T cel cultured alone group (3.10±1.60 vs. 0.94± 0.01, P<0.05). Furthermore, MyD88 in the co-culture group was significantly higher than that in CD4+T cel cultured alone group (2.29±0.05 vs. 1.85±0.31, P<0.01). Tol-like receptor 3 may be involved in the immunoregulation of Th17 cel s by embryonic bone marrow mesenchymal stem cel s, which provides experimental evidence for potential cel therapeutic strategy.

Objective To evaluate the clinical outcome in the treatment of humerus shaft comminuted fractures using limited open reduction and internal fixation combined with an external fixator.Methods Data of 80 patients with comminuted humerus shaft fractures treated from January 2005 to January 2013 were analysed retrospectively.All the patients underwent limited open reduction and internal fixation combined with an external fixator (treatment group) and open reduction and plate fixation (control group) according to the random number table.In the treatment group,there were 40 patients (28 males,12 females),at mean age of 33.5 years (range,21-54 years),with causes of injury including traffic accidents in five patients,falls in nine,crashes in seven and others in six.There were seven patients with open fractures and 33 with closed fractures.In the control group,there were 40 patients (25 males,15 females),at mean age of 32.9 years (range,19-55 years),with causes of injury including traffic accidents in 16 patients,tumbling in seven,crush in seven and others in ten.There were eight patients with open fractures and 32 with closed fractures.The operation time,intraoperative blood loss,bone union time and complications in both groups were recorded.Clinical efficacy was evaluated using the Stewart and Hundley standard.Results Mean follow-up was 19 months (range,15-24 months).Treatment and control groups showed significant differences in operation time [(55.5 ± 10.3) minutes vs.(120.5 ± 15.3) minutes],intraoperative blood loss [(120.4 ± 20.7) ml vs.(245.4 ± 26.7) ml] and bone union time [(11.6 ± 1.3) weeks vs.(14.9 ± 2.3) weeks] (P ＜ 0.05).Rate of incision infection was 8％ (3/40) in treatment group and 10％ (4/40) in control group (P ＞ 0.05).In treatment group the results were excellent in 31 patients and good in nine.In control group the results were excellent in 27 patients,good in nine,fair in one and poor in three.One patient with radial nerve injury after a second surgery for implant removal and two patients with osteomyelitis or bone nonunion were noted in control group.Conclusion Limited open reduction and internal fixation in combination with an external fixator is associated with small trauma,easy operation,short operation time,few bleeding,rigid fixation,early functional exercises and reduced bone nonunion for treatment of comminuted humerus shaft fractures,which exhibits great clinical value.

Objective:To investigate the wear characteristics of bovine enamel and lithium disilicate glass ceramic under simulated oral environment.Methods:18 cylindrical lithium disilicate glass-ceramic specimens with the length of 8 mm and diameter of 3 mm were randomly divided into 2 groups (n =9),9 lithium disilicate glass-ceramic specimens and 9 bovine enamel specimens were served as the antagonists respectively.The specimens were then loaded in a wear simulator and subjected to friction force of 10 N for 540 000 cycles in artificial saliva and room temperature(speed 100 r/min,turning radius of 2.5 mm,uniform circular motion) condition.During the testing,10 checkpoints were applied to measure the height loss of the specimens with 3D profilometer,then wear curves were plotted.Scanning electron microscopy were applied to investigate the worn surfaces at different wear stages.Results:At every checkpoints,bovine enamel wear height loss was larger than the lithium disilicate specimens (P ＜ 0.05);bovine enamel wear curve exhibits a runningin period,steady wear period and severe wear period 3 stages of wear,while wear curves of lithium disilicate glass ceramics exhibit onlyrunning-in period and steady wear period 2 wear stages.Both groups had the corresponding micro-morphological features in different periods.Conclusion:Bovine enamel and lithium disilicate glass ceramics show a phase dynamic evolution law under the simulated oral environment.Bovine enamel is more susceptible to wear than lithium disilicate,suggesting that clinical attention should be paid to prevent the excessive wear of natural teeth caused by lithium disilicate glass ceramic restorations.

Objective:To investigate the aberrant methylation of CpG island in 5′promoter region of p16 gene in the pancreatic juice and its value in diagnosis of patients with pancreatic cancer.Methods:Pure pancreatic juice(PPJ)was collected from the pancreatic duct by a nasopancreatic tube put under endoscopic retrograde cholangiopancreatography(ERCP). Cytological examination was performed by H-E staining in pure pancreatic juice.Aberrant p16 methylation was detected using the methylation specific PCR(MSP)in the PPJ.Results:The sensitivity,specificity,positive predictive value,negative predictive value and accuracy cytological examination in diagnosis of pancreatic cancer were 40%,100%,100%,45.4% and 60.0%,respectively.The DNA sequences were successfully extracted from the PPJ of 30 patients with pancreatic diseases and were subjected to MSP.Seven(35%)of the 20 cases with pancreatic cancer showed aberrant methylation of p16 gene.No aberrant methylation was detected in the pancreatic juice samples of patients with chronic pancreatitis and mucinous cystoadenocarcinoma of pancreas.When cytological examination combined with p16 methylation detection,the sensitivity, specificity,positive predictive value,negative predictive value and accuracy for diagnosis of pancreatic cancer were 55%,100%, 100%,52.6% and 70%,respectively.Conclusion:Pancreatic juice collected by nasopancreatic drainage during ERCP can be used for molecular analysis.Detection of aberrant methylation of p16 gene in pancreatic juice combined with cytological examination is a better method for diagnosis of pancreatic cancer.

PURPOSE: The aim of this study was to investigate the association of telomere length with breast cancer risk. We simultaneously explored the association between telomerase reverse transcriptase gene polymorphisms and telomere length. METHODS: We used real-time quantitative polymerase chain reaction to measure relative telomere length (RTL) in genomic DNA extracted from peripheral blood from 183 breast cancer cases and 191 healthy controls. Genotyping was performed using the Sequenom MassARRAY platform. RESULTS: Our results show that breast cancer patients had significantly shorter RTLs than control subjects (p < 0.05). When the RTLs were categorized into tertiles, we found that the lowest RTL was significantly associated with increased breast cancer risk compared with the highest RTL (odds ratio [OR], 2.33; 95% confidence interval [CI], 1.40–3.90; p=0.001). Subgroup analyses indicated that risk of breast cancer was also significantly increased in the lowest RTL compared with the highest RTL in age >40 years (OR, 2.41; 95% CI, 1.31–4.43; p=0.005), body mass index ≤24 kg/m2 (OR, 2.81; 95% CI, 1.55–5.10; p=0.001), and postmenopausal women (OR, 3.94; 95% CI, 1.63–9.51; p=0.002), respectively. In addition, individuals with the AA genotype of rs2853677 have longer telomeres than those of breast cancer patients with the AG genotype (p=0.011). CONCLUSION: Our results suggest that shorter RTL was associated with an increased risk of breast cancer. An association was found between the AA genotype of rs2853677 and longer RTLs in the case group. Functional studies are warranted to validate this association and further investigate our findings.
Asian Continental Ancestry Group* ; Body Mass Index ; Breast Neoplasms* ; Breast* ; Case-Control Studies* ; DNA ; Female ; Genotype ; Humans ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Telomerase ; Telomere*

Objective:To evaluate the effect of perioperative application of hydroxychloroquine on the prognosis of patients undergoing cardiac surgery.Methods:All SLE patients in the Department of Cardiovascular Surgery of the First Affiliated Hospital of Zhengzhou University who took hydroxychloroquine and glucocorticoid for more than 7 days before operation were enrolled in the observation group(28 cases), including 3 males and 25 females, aged(38.3±8.2)years old. Patients who did not use hydroxychloroquine but only used glucocorticoid before operation were included in the control group(24 cases), including 2 males and 22 females, aged(37.9 ±9.8)years old. There was no significant difference in preoperative clinical data between the two groups in terms of sex, age, BMI, course of systemic lupus erythematosus, hemoglobin, albumin, C-reactive protein, ALT, serum creatinine, ejection fraction, diabetes, hypertension, hyperlipidemia, smoking, alcoholism, preoperative atrial arrhythmia, ventricular arrhythmia, atrioventricular block and so on. The constituent ratio of preoperative operation plan was basically the same between the two groups. The postoperative complications and survival of the two groups were compared.Results:There was no significant difference in early clinical indexes between the two groups, such as cardiopulmonary bypass time( t=0.12, P=0.19), chest drainage volume( t=0.30, P=0.77), second thoracotomy hemostasis( χ2=1.17, P=0.46). There was no significant difference in drug-related complications such as new retinopathy, myocardial concentric hypertrophy, atrial arrhythmia( χ2=1.27, P=0.26), ventricular arrhythmia( χ2=0.98, P=0.32), atrioventricular block( χ2=0.06, P=0.82) and other drug-related complications between the observation group and the control group. There was no significant difference between the two groups in postoperative acute heart failure( χ2=1.17, P=0.28), acute liver insufficiency( χ2=1.17, P=0.28), sternal infection and IABP use( χ2=0.47, P=0.50). Compared with the control group, the incidence of acute renal insufficiency after operation was lower in the observation group( χ2=4.51, P=0.04). The incidence of new postoperative pneumonia was lower( χ2=8.26, P=0.01). The length of postoperative antibiotic use, the length of postoperative ICU hospital stay, the postoperative hospital stay and the total cost of hospitalization in the observation group were significantly less than those in the control group( z=2.71, 2.09, 2.02, 2.02, P=0.01, 0.04, 0.04, 0.04). Compared with the control group, the in-hospital mortality rate of patients in the observation group was lower than that in the control group(3.6% vs. 12.5%, χ2=0.47, P=0.50), and the 6-month and 1-year survival rates of the observation group were higher than those of the control group(92.9% vs.83.3%, 92.9% vs.79.2%; χ2=0.41, 2.17; P=0.53, 0.34), but the difference was not statistically significant. Conclusion:Perioperative administration of hydroxychloroquine can significantly reduce the incidence of postoperative acute renal insufficiency and pneumonia, reduce the duration of postoperative antibiotic use, postoperative ICU hospital stay, postoperative hospital stay, and the cost of hospitalization. Hydroxychloroquine may reduce the in-hospital mortality and improve the long-term survival rate after cardiac surgery, but long-term large sample clinical studies are still needed.

Urinary tract infection (UTI) is one of the most common infectious diseases. It has the characteristics of high recurrence rate and prolonged course. At present, the problem of antibiotic resistance is becoming more and more serious, the incidence of adverse reactions is high, and the disadvantages of long-term administration appear, which brings severe challenges to the treatment of recurrent urinary tract infection. The prevention and treatment of UTI recurrence has become the focus of research. Recurrent urinary tract infection is related to the immune regulation mechanism of the body. Administration of immune regulation can provide new ideas for prevention and treatment. The vaccine based on immune regulation to prevent rUTI has made some progress. It can not only reduce the frequency of recurrences, but also decrease related symptoms. At the same time, the vaccine has good tolerance, high safety and good application prospect. This paper aims to summarize the progress of immune regulation and immune vaccines in vivo and clinical research.

Objective To investigate the role of cardiac rehabilitation program in the early recovery after minimally invasive incision coronary artery bypass grafting with general anesthesia. Methods A retrospective study was performed on the patients who underwent minimally invasive incision coronary artery bypass grafting from January 2015 to January 2020 with general anesthesia in our hospital. The patients were divided into a cardiac rehabilitation group and a control group. The clinical data of the patients were collected in 6 months and 12 months after the beginning of cardiac rehabilitation program and were analyzed by propensity score-matching analysis with a ratio of 1∶1. The main outcomes were the peak oxygen uptake (VO2 peak) of cardiopulmonary function test and the number of patients attending cardiovascular specialties in tertiary hospitals during the follow-up period (20 months). Results A total of 600 patients were enrolled, including 200 patients in the cardiac rehabilitation group [137 males and 63 females, aged 61.00 (56.00, 65.00) years] and 400 patients in the control group [285 males and 115 females, aged 60.00 (56.00, 65.00) years]. After matching, 176 patients were included in each group, and the basical clinical data of the pateints were comparable (P>0.05). The VO2 peak of the cardiac rehabilitation group after 6 months and 12 months of cardiac rehabilitation was significantly different from that of the control group [6 months: 1.96 (1.59, 2.38) L/min vs. 1.72 (1.38, 2.12) L/min, P<0.001; 12 months: 2.40 (2.21, 2.63) L/min vs. 2.12 (1.83, 2.45) L/min, P<0.001]. During the follow-up period, there was a statistical difference in the cardiovascular specialist visits in tertiary hospitals (P=0.004). Conclusion Cardiac rehabilitation program has a positive effect on the recovery of minimally invasive incision coronary artery bypass grafting with general anesthesia, and can improve the exercise ability of patients.

Aging-elevated DNMT3A R882H-driven clonal hematopoiesis (CH) is a risk factor for myeloid malignancies remission and overall survival. Although some studies were conducted to investigate this phenomenon, the exact mechanism is still under debate. In this study, we observed that DNMT3A R878H bone marrow cells (human allele: DNMT3A R882H) displayed enhanced reconstitution capacity in aged bone marrow milieu and upon inflammatory insult. DNMT3A R878H protects hematopoietic stem and progenitor cells from the damage induced by chronic inflammation, especially TNFα insults. Mechanistically, we identified that RIPK1-RIPK3-MLKL-mediated necroptosis signaling was compromised in R878H cells in response to proliferation stress and TNFα insults. Briefly, we elucidated the molecular mechanism driving DNMT3A R878H-based clonal hematopoiesis, which raises clinical value for treating DNMT3A R882H-driven clonal hematopoiesis and myeloid malignancies with aging.