ObjectiveTo investigate the effect of nicotinamide mononucleotide adenosyl transferase 3 (NMNAT3) on the mitochondrial function and anti-oxidative stress of rabbit bone marrow mesenchymal stem cells (BMSCs) under oxidative stress in vitro by regulating nicotinamide adenine dinucleotide (NAD+) levels.MethodsThe bone marrow of femur and tibia of New Zealand white rabbits were extracted. BMSCs were isolated and cultured in vitro by density gradient centrifugation combined with adherent culture. The third generation cells were identified by flow cytometry and multi-directional induction. Overexpression of NMNAT3 gene was transfected into rabbit BMSCs by enhanced green fluorescent protein (EGFP) labeled lentivirus (BMSCs/Lv-NMNAT3-EGFP), and then the expression of NMNAT3 was detected by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot and cell proliferation by cell counting kit 8 (CCK-8) method. BMSCs transfected with negative lentivirus (BMSCs/Lv-EGFP) and untransfected BMSCs were used as controls. The oxidative stress injury cell model was established by using H2O2 to treat rabbit BMSCs. According to the experimental treatment conditions, they were divided into 4 groups: Group A was normal BMSCs without H2O2 treatment; untransfected BMSCs, BMSCs/Lv-EGFP, and BMSCs/Lv-NMNAT3-EGFP in groups B, C, and D were treated with H2O2 simulated oxidative stress, respectively. The effects of NMNAT3 on the mitochondrial function of BMSCs under oxidative stress [changes of mitochondrial membrane potential, NAD+ and adenosine triphosphate (ATP) levels], the changes of anti-oxidative stress ability of BMSCs [reactive oxygen species (ROS) and malondialdehyde (MDA) levels, manganese superoxide dismutase (Mn-SOD) and catalase (CAT) activities], and the effects of BMSCs on senescence and apoptosis [senescence associated-β-galactosidase (SA-β-gal) staining and TUNEL staining] were detected after 24 hours of treatment.ResultsThe rabbit BMSCs were successfully isolated and cultured in vitro. The stable strain of rabbit BMSCs with high expression of NMNAT3 gene was successfully obtained by lentiviral transfection, and the expressions of NMNAT3 gene and protein significantly increased (P<0.05). There was no significant difference in the trend of cell proliferation compared with normal BMSCs. After treatment with H2O2, the function of mitochondria was damaged and apoptosis increased in all groups. However, compared with groups B and C, the group D showed that the mitochondrial function of BMSCs improved, the membrane potential increased, the level of NAD+ and ATP synthesis of mitochondria increased; the anti-oxidative stress ability of BMSCs enhanced, the levels of ROS and MDA decreased, and the activities of antioxidant enzymes (Mn-SOD, CAT) increased; and the proportion of SA-β-gal positive cells and the rate of apoptosis decreased. The differences in all indicators between group D and groups B and C were significant (P<0.05).ConclusionNMNAT3 can effectively improve the mitochondrial function of rabbit BMSCs via increasing the NAD+ levels, and enhance its anti-oxidative stress and improve the survival of BMSCs under oxidative stress conditions.
ObjectiveTo study the effects of the new small molecular oxygen free radical scavenger Tempol on the survival and vasculogenesis of the long random pattern skin flap (LRPSF) and its mechanism. MethodsEighty-four male Sprague Dawley rats were randomly divided into control and Tempol groups (42 rats in each group). LRPSF of 9 cm×3 cm in size were prepared on the backs of rats in two groups based on the Mcfarlane flap. Rats were administered with Tempol (100 mg/kg) in the Tempol group and with normal saline in the control group by intraperitoneal injection at 15 minutes before operation and at 1-7 day after operation. The rat and the skin flap survival conditions were observed after operation; the survival rate of skin flap was measured, and the vascular structure, vascular volume, and total length of blood vessels were analyzed with Micro-CT three-dimensional imaging after 7 days; HE staining was used to observe the structure of the skin flaps and inflammation, immumohistochemical staining to observe vascular endothelial growth factor (VEGF) expression; water-soluble tetrazolium-1 method was used to measure the content of superoxide dismutase (SOD) and malondialdehyde (MDA), and ELISA to detect the expressions of tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) after 1, 3, and 7 days. ResultsAll of rats survived after operation, without hemorrhage, edema, and infection. With the extension of time, necrosis occurred in the distal part of the skin flaps in 2 groups, but the necrosis degree of the Tempol group was lower than that of control group; meanwhile, the blood vessel distribution and continuity were better than those of control group. The skin flaps survival rate, vascular volume, and total length of blood vessels of Tempol group were significantly higher than those of control group after 7 days (P<0.05). The clearer skin flaps structure, lighter inflammation reaction and inflammation cell infiltration, and higher VEGF staining intensity were observed in the Tempol group than the control group after 7 days. There was no significant difference in SOD, MDA, and TNF-α, and IL-6 contents between the 2 groups at immediate after operation. SOD significantly increased, but MDA, TNF-α, and IL-6 contents significantly decreased in the Tempol group when compared with control group after 1, 3, and 7 days (P<0.05). ConclusionTempol can significantly promote the LRPSF survival rates, its mechanism is closely related to the promotion of vasculogenesis and reduction of oxidative stress and inflammation.
Objective To investigate the correlation between stress hyperglycemia ratio (SHR) and acute ischemic stroke (AIS) 1-year prognosis, to provide more clinical basis to improve the prognosis of AIS patients and to target and control the influencing factors. MethodsThe patients with AIS diagnosed for the first time and received treatment at the Shijiazhuang Fifth Hospital between May 2019 and January 2022 were retrospectively and continuously included. According to the Modified Rankin Scale score 1-year after the onset of the disease, the patients were divided into a good prognosis group and a poor prognosis group. Also the patients were divided into 2 groups based on the median of SHR. The correlation between SHR and stress blood glucose was analyzed, and the factors affecting the prognosis of AIS patients were identified. The predictive value of SHR and stress blood glucose on the prognosis of AIS patients was compared using receiver operating characteristic. Results A total of 206 patients were included. Among them, there were 125 cases (60.7%) in the good prognosis group and 81 cases (39.3%) in the poor prognosis group. The median SHR (lower quartile, upper quartile) is 1.20 (1.08, 1.33). There were statistically significant differences between the two groups in the scores of the National Institutes of Health Stroke Scale, diabetes history, hypertension history, low-density lipoprotein cholesterol, stress blood glucose, age, SHR and SHR classification (P<0.05). There was no statistically significant difference in the other indicators compared between the two groups (P＞0.05). Stress blood glucose was positively correlated with SHR (7.95±1.78 vs. 1.21±0.19; r=0.294, P<0.001). Multivariate logistic analysis showed that stress blood glucose and SHR were independent factors influencing the 1-year prognosis of AIS patients (P<0.05), and the interaction between SHR and diabetes was not significant (P>0.05) After adjusting for confounding factors, the area under the receiver operating characteristic curve of SHR for the prognosis of AIS patients was higher than that of stress blood glucose [0.682 (0.614, 0.745) vs .0.585 (0.515, 0.653); Z=2.042, P=0.041]. Conclusions SHR and stress blood glucose are independent risk factors for 1-year prognosis in AIS patients. However, SHR has a better predictive value for 1-year prognosis in AIS patients than stress blood glucose. Whether the patient has diabetes or not, the impact of SHR on the prognosis of AIS patients is consistent.
Objective To assess the clinical efficacy and safety of proton pump inhibitor （PPI） and H2RA for stress ulcer bleeding in stroke patients. Methods Randomized controlled trials （RCT） were identified from MEDLINE （ 1966- Oct. 2005 ） ,EMBASE （ 1984- Oct. 2005 ）, The Cochrane Library （ Issue 4,2005 ）, CBMdisc （ 1980- Oct. 2005 ） and VIP（ 1980- Oct. 2005 ）. We handsearched the related published and unpublished data and their references. The quality of included trials was evaluated. Data were extracted by two reviewers independently with a designed extraction form. RevMan 4. 2.7 software was used for data analysis. Results Twenty RCT were included with 2 624 patients. The results of meta-analysis were listed as follows： （1） stress ulcer bleeding （SUB） ： PPI （ OR 0.14,95% CI 0.08 to 0.24, NNT = 3 ） and H2RA （OR 0.24,95% CI 0.15 to 0.39, NNT =5） significantly reduced the incidence of SUB in comparison with control group. PPI significantly reduced the incidence of SUB compared with H2R.A（P 〈0. 00001 ）. （2） Mortality： PPI （OR 0.22,95% CI 0. 11 to 0.47, NNT =8） and H2RA （OR 0.53,95% CI 0. 34 to 0.81, NNT =16） significantly decreased the mortality compared with non-prophylaxis group. PPI significantly decreased the mortality compared with H2RA （OR 0.28,95% CI 0.09 to 0. 89）. （3） Adverse effect： There were not evident adverse effects in both PPI and H2RA groups. Conclusions PPI and H2RA may reduce the incidence and mortality of SUB in stroke patients, and PPls are better in reducing incidence of SUB than H2RA.
目的 了解5.12汶川大地震24个月后震区小学生的心理健康状况，为进行震后长期心理危机干预提供依据。 方法 于2010年5月即汶川大地震发生后24个月，分别使用创伤应激量表儿童版（CRIES-13）、儿童抑郁障碍自评量表（DSRSC）、长处与困难问卷（SDQ）学生版，调查了553名来自于彭州灾区的小学生的心理健康状况及其影响因素。 结果 ① 灾区小学生CRIES-13总分为（22.98 ± 12.29）分，其中大于划界分（30分）者为143人，占总人数的25.9%；女性总分高于男性（Z=?2.031，P=0.042）；震后被转移至安全地点的时间越长（OR=1.025，P=0.012）、家人伤亡越严重（OR=1.141，P=0.021），其CRIES-13总分大于划界分的可能性越高。② 灾区小学生DSRSC总分为（11.07 ± 5.78）分，其中总分大于划界分（15分）者为120人，占总人数的21.7%；女性总分高于男性（Z=?2.508，P=0.012）；灾区小学生的年龄（r=0.098，P=0.021）、震后被转移至安全地点的时间（r=0.117，P=0.004）与DSRSC总分呈正相关。③ 灾区小学生SDQ总困难因子得分为（14.97 ± 5.44）分，62.9%的人报告自身存在主观困难；女性的情绪因子、亲社会因子得分高于男性（Z=?3.123，P=0.002；Z=?2.243，P=0.025）；年龄越大，品行因子（χ2=7.604，P=0.023）、亲社会因子（χ2=8.102；P=0.017）得分增加。 结论 震后灾区小学生的心理健康状况受到性别、年龄、震后被转移至安全地点的时间、家人伤亡程度的影响，震后长期心理危机干预应综合考虑这些相关因素以确定高危人群。
Objective To observe the effects of cigarette smoke extract ( CSE) on the proliferation and secretion of hydrogen peroxide ( H2O2 ) in human lung fibroblasts ( HLFs) induced by transforming growth factor-β1 ( TGF-β1 ) . Methods Cultured HLFs were divided into a normal group and a model group induced by TGF-β1 ( 5 ng/mL) , then intervened with CSE at different concentrations ( 0% , 2. 5% , 5% ,10% , respectively) . Brdu ELISA assay was used to detect cell proliferation. H2O2 release from cultured cells was assayed using a fluorimetric method. Cellular localization of H2O2 and expression of α-SMA were performed using a fluorescent-labeling strategy. Results TGF-β1 stimulated group showed positive expression of α-SMA, implying TGF-β1 had induced fibroblasts to differentiate into myofibroblasts. In TGF-β1 stimulated group, 2. 5% and 5% CSE promoted cell proliferation ( P lt; 0. 01 or 0. 05) , while 10% CSE inhibited cell proliferation ( P lt; 0. 01) . In the normal group, both low and high concentration of CSE inhibited cell proliferation ( P lt; 0. 01 or P lt; 0. 05) , and the inhibition effect was dose-dependent. HLF induced by TGF-β1 generated low constitutive levels of extracellular H2O2 that was markedly enhanced by CSE treatment ( P lt; 0. 01) . Pretreatment with DPI, an inhibitor of NADPH oxidase, abolished secretion of H2O2 . Cellular localization of H2O2 by a fluorescent-labeling strategy demonstrated that extracellular secretion of H2O2 is specific to the myofibroblast. Conclusions Low concentration of CSE can promote myofibroblast proliferation, and markedly increase extracellular secretion of H2O2 . CSE possibly take part in the development and progress of idiopathic pulmonary fibrosis by increasing oxidative stress.
Objective To investigate the quality of life (QOL) of Beichuan cadres 1 year after Wenchuan earthquake, so as to provide references for promoting psychological intervention and improving QOL. Methods With the adoption of WHOQOL-BREF, questionnaire survey on basic conditions of cadre victims, and PCL-C scale, cross-section survey was conducted for 2 055 cadres of Beichuan public institutions in aspects of their QOL, social demographic characteristics and psychological trauma. Results a) Compared with domestic norm, although the Beichuan cadres had higher score in social relation domain (14.31 ± 2.59, Plt;0.01), their QOL scores in each dimensions (physical domain score: 14.16 ± 2.43; psychological domain score: 13.79 ± 2.59, environmental domain score: 11.20 ± 2.74) were lower with significant difference (Plt;0.01); b) The group with population under the age of 30 scored higher than other age groups in the psychological and environmental domains; men scored higher than women in the psychological domain; the widowed group scored lower than other groups in all domains; the Han nationality group scored higher than the Qiang nationality group in all domains; the primary education group scored lower than other groups in the social relation domain; the group with relatives, houses and physical loss or damage scored lower than the non-loss group in all domains; and the higher the score in PCL-C, the lower the scores in all domains (all Plt;0.05). Conclusion Beichuan cadres are severely affected in their physical, psychological and environmental domains even one year after Wenchuan earthquake, especially the female, the Qiang nationality, the old, the lost, and the psychological injured cadres are more badly affected with worse QOL, so those population should be concerned in particular with sustainable psychological service and other measures. The finding of social relation domain score being higher than general population suggests that it is important to promote the satisfaction with social relation through the timely effective support from both government and civil society, and the resettlement of sample housing area as well.
Objective To observe the protein expression of c-Jun amino-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) in normal skin and keloid and to explore their influences on the formation of kloid. Methods Keloid tissues and normal skin tissues were collected from 16 keloid resection patients (experimental group) and 10 voluntary plastic surgery patients (control group). In the experimental group, the keloid formation time ranged from 8 months to 10 years; the keloid tissues were collected from the chest in 6 cases, the ear lobe in 4 cases, the perineum in 2 cases, the shoulder in 3 cases, and the abdomen in 1 case; and all keloid tissues were confirmed by pathological examination. In the control group, normal skin tissues were collected from the abdomen in 4 cases, the thighs in 3 cases, the shoulder in 2 cases, and the back in 1 case. Two-step l ine of Envision immunohistochemical staining was performed to observe the expressions of nonphosphorylated and phosphorylated JNK and ERK; Image Pro Plus 4.5 image analysis system was used to measure the integrated absorbance (IA) and to observe the positive staining strength. Results The immunohistochemical staining showed that no obvious expressions of phosphorylated and non-phosphorylated ERK, JNK were observed in the fibroblasts of the control group, and the expressions of phosphorylated JNK and ERK proteins were significantly higher in the experimental group than in the control group (P lt; 0.05). There was no significant difference in the expressions of non-phosphorylated JNK and ERK proteins between 2 groups (P gt; 0.05). Conclusion Activation of ERK and JNK pathways might be involved in formation of keloid.