Objective: To observe the expression of long noncoding RNA (LncRNA) AK009271 in the cortex of rats with cerebral ischemia-reperfusion,and to evaluate the effect of selective brain hypothermia on the its expression. Methods: All 75 healthy male Sprague-Dawley (SD) rats of SPF grade, weighing 200-250 g,were randomly divided into the sham operation group (group S), the ischemia-reperfusion group (group I/R) and the hypothermia group (group HT) by random digital scale method,with 25 rats in each group. Focal cerebral ischemia-reperfusion model was induced by string Longa's thread thrombus method of the middle cerebral artery for 2 hours. The group HT was infused with 4°C cold isotonic saline for 15 min (20 ml/kg) through internal carotid artery immediately after reperfusion. During the procedure, brain and rectal temperatures were monitored to ensure successful construction of selective brain hypothermia model. The group S only separated the carotid artery without performing middle cerebral artery embolization. At 6,24,and 48 h after reperfusion, the neurological deficit scores were measured and the expression of LncRNA AK009271 was evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). At 24 h after reperfusion, the expression of Fisl was determined by Western Blot, and mitochondrial morphology was observed by electron microscopy. The apoptosis rate of cells was detected by TUNEL staining. Results: (1) Compared with the group I/R, the group HT had decreased brain temperature after perfusion of 4 °C isotonic saline for 15 min (32. 27 ± 0. 15 vs. 34. 52 ± 0. 22,t = 1. 603, P < 0. 05), but without significant changes in rectal temperature (P > 0. 05). (2) Compared with the group S,the neurological deficit scores were increased in the other two groups;the neurological deficit score in the group HT was lower than the group I/R; all P < 0. 05. (3) Compared with the group S, the expression of LncRNA AK009271 in the group I/R all decreased at 6,24 and 48 h after reperfusion,and reached the lowest at 24 h after reperfusion (all P <0. 05);however,the expression of LncRNA AK009271 in the group HT increased at 6,24 and 48h after reperfusion compared with the group I/R (both P<0.05). (4) Compared with the group S, up-regulated expression of Fisl, mitochondrial ultrastructure damage and increased apoptosis rate were observed in the other two groups at 24 h after reperfusion (all P < 0. 05); However, compared with group I/R, down-regulated expression of Fisl (P <0. 05), less damaged mitochondrial ultrastructure and decreased apoptosis rate were observed in the HT group (P < 0. 05). Conclusions The expression of LncRNA AK009271 in the cortex of rats with cerebral ischemia-reperfusion is decreased. Selective brain hypothermia could ameliorate focal cerebral I/R injury in rats through up-regulating LncRNA AK009271 expression, inhibiting Fisl expression and lightening cell apoptosis.

Objective To investigate the relationships between low one-minute Apgar score and the prognosis of extremely preterm infants (EPI) and extremely low birth weight infants (ELBWI). Methods Altogether 50 EPI and ELBWI who had a low one-minute Apgar score ( ≤ 7) and were admitted to the Neonatal Intensive Care Unit (NICU) of Peking University Third Hospital from January 1,2010 to December 31, 2015 were enrolled in this study. All of them were divided into two groups according to their Apgar score: mild group (4-7) and severe group (0-3). Medical records of the subjects were reviewed and an at least 18 months follow up study was conducted. Conditions of all subjects during perinatal period and hospitalization were summarized. Outcomes and follow-up results were compared between the two groups by using Fisher exact test. Results (1) General information: Fifty infants were involved, among which 37 had a mild low Apgar score and 13 had a severe low Apgar score. The mean gestational age was (27.7±2.1) weeks and the mean birth weight was (884.4±174.3) grams. (2) Main complications (some infants with more than one complication): There were 42 cases of neonatal respiratory distress syndrome, 12 cases of pulmonary hemorrhage, 21 cases of bronchopulmonary dysplasia, 31 cases of patent ductus arteriosus, 36 cases of intraventricular hemorrhage, 22 cases of white matter damage and six cases of retinopathy of prematurity. (3) Outcomes: The survival rate was 48% (24/50) and the mortality rate was 52% (26/50). Among the 26 infants, five died despite treatment and 21 died within 72 hours after their parents giving up treatment. There were no significant differences in the survival rates, mortality rates and rates of abandon treatment between the two groups [43% (16/37) vs 8/13; 11%(4/37) vs 1/13; 46% (17/37) vs 4/13; Fisher exact test, all P>0.05]. (4) Follow-up results: Twenty-one infants were followed-up to at least 18 months of age, among which four were normal, 10 had growth retardation and recurrent respiratory tract infection and seven had motor development retardation. The incidence of motor development retardation in severe group was higher than that in mild group, and the difference between them was statistically significant (5/8 vs 2/13, Fisher exact test, P=0.046). Conclusions EPI or ELBWI with a low one-minute Apgar score have many nosocomial complications, resulting in high mortality and high incidence of motor development retardation.

Lung cancer is one of the most common malignancies. Anti-tumor drugs with intravenous administration have systemic adverse effects as well as limited efficacy. Drugs can concentrate in lungs after pulmonary administration,which limits the distribution in the other organs and reduces the side effects of anti-tumor drugs. The paper focused on the recent progress in the studies on new dos-age forms of anti-tumor drugs for pulmonary administration for the therapy of lung cancer,so as to provide reference for the development of anti-tumor drugs for pulmonary administration.

Objective:To investigate the role of Sestrin2 overexpression in regulating mitochondrial fission and its mechanism in human neuroblastoma SH-SY5Y cell model of glucose and oxygen deprivation/recovery (OGD/R). Methods:(1) SH-SY5Y cells were divided into normal control group, OGD/R group, Vector group, and Sestrin2 overexpression group; Sestrin2 overexpression or empty vector stable cell lines in the Sestrin2 overexpression group and Vector group were constructed by lentivirus infection; cells in the later 3 groups were subjected to oxygen-glucose deprivation (OGD) for 4 h followed by restoration of O 2 supply for 18 h. The cell survival rate was detected by cell counting kit (CCK)-8 assay. The protein levels of Sestrin2, dynamin-related protein 1 (Drp1), mitochondrial fission protein 1 (Fis1), B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), Kelch-like ECH-related protein 1 (Keap1) in the cytoplasm and nuclear factor E2-related factor (Nrf2) in the nucleus were detected by Western blotting. The mitochondria ultrastructure was observed by transmission electron microscope. The Nrf2 nuclear translocation was detected by immunofluorescence staining. (2) Cell lines with Sestrin2 overexpression were divided into Sestrin2 overexpression group, Brusatol+ Sestrin2 overexpression group, and DMSO+ Sestrin2 overexpression group. Cells in the Brusatol+ Sestrin2 overexpression group were pretreated with normal medium containing Brusatol (Keap1/Nrf2 pathway inhibitor, final concentration: 100 nmol/L) for 4 h before OGD/R; cells in the DMSO+ Sestrin2 group were pretreated with normal medium containing DMSO (final volume fraction: 0.1%) for 4 h before OGD/R. Cells in these groups were then subjected to OGD for 4 h followed by restoration of O 2 supply for 18 h. The protein levels of Drp1, Fis1, Keap1 in the cytoplasm, and Nrf2 in the nucleus were measured by Western blotting. Results:(1) As compared with those in the OGD/R group, cells in the Sestrin2 overexpression group had significantly increased survival rate (61.33%±1.15% vs. 81.00%±3.00%), significantly up-regulated Bcl-2/Bax ratio (0.467±0.006 vs. 0.880±0.010), significantly decreased Drp1, Fis1 and cytoplasmic Keap1 protein levels (1.089±0.033 vs. 0.865±0.014; 0.829±0.009 vs. 0.350±0.007; 0.967±0.017 vs. 0.881±0.024), and significantly up-regulated nuclear Nrf2 protein level (0.627±0.025 vs. 0.957±0.015, P<0.05). The mitochondrial structure in the Sestrin2 overexpression group under electron microscope was more complete than that in the OGD/R group, and obvious nuclear translocation of Nrf2 was noted. (2) As compared with the Sestrin2 overexpression group, Brusatol+ Sestrin2 overexpression group had significantly decreased nuclear Nrf2 protein level (0.920±0.013 vs. 0.627±0.035), and statistically increased Drp1 and Fis1 protein levels (0.994±0.020 vs. 1.084±0.005; 0.728±0.010 vs. 0.906±0.022, P<0.05). Conclusion:Sestrin2 overexpression could suppress mitochondrial fission, reduce cell apoptosis, and attenuate OGD/R injury of SH-SY5Y cells by activating Keap1/Nrf2 pathway via down-regulating cytoplasmic Keap1 protein level and promoting Nrf2 nuclear translocation.

OBJECTIVE: To explore the effect of chitosan (CS) hydrogel loaded with tendon-derived stem cells (TDSCs; hereinafter referred to as TDSCs/CS hydrogel) on tendon-to-bone healing after rotator cuff repair in rabbits. METHODS: TDSCs were isolated from the rotator cuff tissue of 3 adult New Zealand white rabbits by Henderson step-by-step enzymatic digestion method and identified by multidirectional differentiation and flow cytometry. The 3rd generation TDSCs were encapsulated in CS to construct TDSCs/CS hydrogel. The cell counting kit 8 (CCK-8) assay was used to detect the proliferation of TDSCs in the hydrogel after 1-5 days of culture in vitro, and cell compatibility of TDSCs/CS hydrogel was evaluated by using TDSCs alone as control. Another 36 adult New Zealand white rabbits were randomly divided into 3 groups ( n=12): rotator cuff repair group (control group), rotator cuff repair+CS hydrogel injection group (CS group), and rotator cuff repair+TDSCs/CS hydrogel injection group (TDSCs/CS group). After establishing the rotator cuff repair models, the corresponding hydrogel was injected into the tendon-to-bone interface in the CS group and TDSCs/CS group, and no other treatment was performed in the control group. The general condition of the animals was observed after operation. At 4 and 8 weeks, real-time quantitative PCR (qPCR) was used to detect the relative expressions of tendon forming related genes (tenomodulin, scleraxis), chondrogenesis related genes (aggrecan, sex determining region Y-related high mobility group-box gene 9), and osteogenesis related genes (alkaline phosphatase, Runt-related transcription factor 2) at the tendon-to-bone interface. At 8 weeks, HE and Masson staining were used to observe the histological changes, and the biomechanical test was used to evaluate the ultimate load and the failure site of the repaired rotator cuff to evaluate the tendon-to-bone healing and biomechanical properties. RESULTS: CCK-8 assay showed that the CS hydrogel could promote the proliferation of TDSCs ( P<0.05). qPCR results showed that the expressions of tendon-to-bone interface related genes were significantly higher in the TDSCs/CS group than in the CS group and control group at 4 and 8 weeks after operation ( P<0.05). Moreover, the expressions of tendon-to-bone interface related genes at 8 weeks after operation were significantly higher than those at 4 weeks after operation in the TDSCs/CS group ( P<0.05). Histological staining showed the clear cartilage tissue and dense and orderly collagen formation at the tendon-to-bone interface in the TDSCs/CS group. The results of semi-quantitative analysis showed that compared with the control group, the number of cells, the proportion of collagen fiber orientation, and the histological score in the TDSCs/CS group increased, the vascularity decreased, showing significant differences ( P<0.05); compared with the CS group, the proportion of collagen fiber orientation and the histological score in the TDSCs/CS group significantly increased ( P<0.05), while there was no significant difference in the number of cells and vascularity ( P>0.05). All samples in biomechanical testing failed at the repair site during the testing process. The ultimate load of the TDSCs/CS group was significantly higher than that of the control group ( P<0.05), but there was no significant difference compared to the CS group ( P>0.05). CONCLUSION TDSCs/CS hydrogel can induce cartilage regeneration to promote rotator cuff tendon-to-bone healing.
Rabbits ; Animals ; Rotator Cuff/surgery* ; Chitosan ; Hydrogels ; Rotator Cuff Injuries/surgery* ; Wound Healing ; Tendons/surgery* ; Collagen ; Stem Cells ; Biomechanical Phenomena

ObjectiveTo observe the effect of enriched environment (EE) combined with acupuncture at head point (HA) on behavior in rats with autism spectrum disorder. MethodsHealthy female Wistar rats were given peritoneal injection of sodium valproate at 12.5 days of gestation. Twenty-four male offspring rats were randomly selected and then randomly divided into model group (n = 6), EE group (n = 6), HA group (n = 6) and EE combined with HA group (the combined group, n = 6). Six male offspring rats born from female mice injected with the same amount of saline intraperitoneally were as control group. After four weeks of treatment, all the five groups were tested with three-chamber test and marble burying test, and the sociability index, the social novelty index and the number of buried marbles were recorded. The levels of interleukin (IL)-1β and IL-6 in peripheral blood were determined by enzyme-linked immunosorbent assay (ELISA). ResultsAfter treatment, compared with the model group, the sociability index and the social novelty index improved (P < 0.05), the number of buried marbles reduced (P < 0.05), and the levels of IL-6 and IL-1β in peripheral blood decreased in EE group, HA group and the combined group (P < 0.05); while the combined group was the best (P < 0.01). ConclusionBoth EE or acupuncture at HA could improve behavioral symptoms, and reduce the expression of inflammatory factors in rats with autism spectrum disorder. The combination of the two methods showed the best result.