ObjectiveTo investigate the roles of 11 β-hydroxysteroid dehydrogenase type 1 ( 11 β-HSD1 )on hippocampus of rat with chronic unpredictable mild stress (CUMS).MethodsTwenty-four male SpragueDawley rats were randomly divided into control group and depressive model group. Chronic unpredictable mild stress (CUMS) was used to make up depressive animal model.Behavioral changes were recorded by body weight measuring,sucrose consumption test (SCT) and open field test (OFT),respectively.The mRNA transcription of 11β-HSD1 in hippocampus tissues of the rats were detected by real-time RT-PCR,and the protein expression of 11β-HSD1 were detected by western blot and immunofluorescence.ResultsBcforc starting CUMS protocol,the rats exhibited equivalent weight and sucrose consumption.Twenty-eight days after CUMS protocol,behavior parameters such as body weight,sucrose consumption,nunber of crossing,and number of rearing were significantly decreased in rats exposed to CUMS group compared with control group (P ＜ 0.05,P ＜ 0.01 ).Correspondingly,realtime RT-PCR assays showed the mRNA expression of 11 β-HSD1 in the hippocampus of CUMS group,which was (31 ±9) ％ lower than that of control group.Meanwhile,the protein expression of it in CUMS group was lower than that of control group (P ＜ 0.05 ).Inmunofluorescence revealed that the number of positive 11 3-HSD1 cells was high (223 ± 13) in the control group,while the number was decreased prominently (92 ± 11 ) in the CUMS group (P ＜ 0.01 ).ConclusionDepressive behavior of rats is induced and the expression of 11 β-HSD1 in the hippocampus is decreased prominently by CUMS,the mechanism of which is at least related to the low expression of 11β-HSD1 and disturbance of glucocorticoid metabolism caused by CUMS.

Objective To study the effect of brain-derived neurotrophic factor (BDNF) on the environmental nutrition and neural differentiation of the transplanted stem cells under hypothermia.Methods The BDNF gene mediated by liposome was transfected into 293T cell line, and ELISA assay was applied to find the peak time of BDNF expression. When BDNF was highly expressed, the supernatant was collected for establishment of SD rat models of brain injury. The rats were divided into Group A (stem cell transplantation group) and Group B (stem cell transplantation and BDNF group). Rats in both groups were under hypothermia treatment for five days. Four and eight days later ( three days from rewarming), rat brain tissues were obtained to detect the expressions of proliferating cell nuclear antigen (PCNA), nestin, neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) by immunohistochemical method and to detect the apoptosis by in situ hybridization. Finally, the nerve function scores were obtained for evaluation of the nerve function. Results The ELISA showed that the high level of BDNF expression was at 48 to 60 hours after gene transfection. PCNA and nestin were highly expressed, while NES and GFAP showed nil or low level of expression in both groups at the fourth day after hypothermia, with little apoptotic cells especially in the Group B (P ＜0.05). The expressions of PCNA and nestin were decreased, but the expressions of NSE and GFAP were increased at the third day after rewarming. The positive rate of NSE expression in the Group B was much higher and the apoptotic cells were much less compared with the Group A ( P ＜ 0. 05 ). A better nerve score was obtained in the Group B. Conclusion BDNF can enhance the survival rate of the transplanted stem cells and induce their differentiation into neurons under hypothermia.

Objective To study the pulmonary surfactant (PS) on prevention of neonatal respiratory distress syndrome (NRDS) in neonates delivered via caesarean section. Methods From selective cesarean section infants (gestational age 34-38+6 W), 80 cases whose test tube oscillation tests were negative and amniotic fluid pulmonary surfactant associated protein A (SP-A) concentrations were lower than <10μg/L, and were randomly divided into PS prevention group and control group, with 40 cases in each group. PS prevention group within 1 h of birth were administrated poractant alfa injection by endotracheal tube (dose 100 mg/kg), but the control group was not given special treatment, leaving only the observation. The incidence of NRDS, treatment status and clinical progression were compared between two groups. Results The incidence of NRDS in control group was 82.5%(33/40), in PS prevention group was 37.5%(15/40), and there was significant difference (P<0.05). The degree of NRDS in control group was more severe. The incidence rate of persistent pulmonary hypertension of the new-born (PPHN), pulmonary air leak, patent ductus arteriosus and oxygenation index above 25 mmHg (1 mmHg=0.133 kPa) in control group were significantly higher than those in PS prevention group (P<0.05). The time of mechanical ventilation, the time of oxygen inhalation, ratio of arterial partial pressure of oxygen (PaO2) before mechanical ventilation to fraction of inspired oxygen (FiO2), and costs of hospitalization in control group were significantly higher than those in PS prevention group (P<0.05). Conclusions PS prevention can reduce the incidence of NRDS of neonates delivered by elective caesarean section, can alleviate the symptoms of NRDS, shorten length of stay and reduce the cost of hospitalization.

Objective To evaluate the effect of Saccharomyces boulardii (SB) administration on very-low-birth-weight (VLBW) infants.Methods One hundred and ninety-eight preterm infants were prospectively randomized into observation group (105 cases) and control group (93 cases) based on the symptomatic and supportive treatment.When uncompletely stomach intestine nutrition fed,the patients of observation group took SB (50 mg/kg),the patients of control group took equivalent placebo.The times of defecation and diarrhea,the rate of neonatal necrotizing enterocolitis,hospital onset of infection (septicemia,pulmonary infection),fungal infection,the time of intravenous nutrition and length of stay were compared.Results The general data in two groups had no significant difference (P ＞ 0.05).The times of defecation,time of intravenous nutrition and length of stay in two groups had significant difference [(1.8 ± 0.4) times/d vs.(3.4 ± 0.5) times/d,(30.21 ± 3.43) d vs.(40.47 ± 4.35) d,(33.5 ± 6.8) d vs.(45.4 ± 9.3) d] (P ＜ 0.05).The rate of diarrhea,neonatal necrotizing enterocolitis,septicemia and pyemia in two groups had significant difference [14.3％ (15/105) vs.25.8％ (24/93),11.4％ (12/105) vs.19.4％ (18/93),19.0％ (20/105) vs.29.0％ (27/93)] (P ＜ 0.05).The rate of pulmonary infection and fungal infection between two groups had no significant difference(P＞ 0.05).Conclusion SB administration on VLBW infants can reduce the infection,promote enteral feeding,shorter hospital stay,and has a certain significance on the family and the community.

Objective To establish the electric controlled cortical impact (eCCI)-induced traumatic brain injury (TBI) model in rats with different severity in degree,which may serve as a suitable platform to provide experimental evidence for the pathophysiological following TBI.Methods A total of 40 male Wistar rats were randomly divided into 3 experimental groups and sham group.TBI rats (n=10/group) were positioned beneath the controlled cortical impactor device (eCCI) and subjected to impact injury at 2 mm depth of penetration,for a sustained depression of 200 ms,at 4 m/s,5 m/s,6 m/s velocity for mild,moderate,and severe TBI,respectively.Sham-operated rats (n=10) underwent identical surgical procedures,including craniotomy,without receiving the cortical impact.Neurological function and regional cerebral flow (24 h after CCI),contusion volume,histopathological,and ultrastructural changes (48 h after CCI) were measured,respectively.Results The severity of the pathological changes in rats was increased as the injury aggravated.The eCCI device impacted the brain at 4 m/s,5 m/s,6 m/s velocity for mild,moderate,and severe TBI,respectively.TBI groups showed impaired neurological function,and decreased rCBF lower than that of sham-operated group (all P＜0.01).Furthermore,neuronal pathological abnormalities in TBI groups,including neuron shrinking,perineuronal vacuole,and structural abnormalities of mitochondria.Increased severity of injury was apparent following the increased level of the impacted velocity,and significant differences were observed between TBI groups (P＜0.05).Conclusion The TBI animal model with mild,moderate,and severe brain injury can be established successfully by 4 m/s,5 m/s,and 6 m/s of impact velocity respectively with the eCCI-6.3 device.The novel eCCI-induced TBI model in rats possibly serves as a novel useful approach in the development of TBI models.

Objective To explore the relationship between plasma protein Z level and severity of coronary atherosclerosis in patients with coronary heart disease,and analyze the clinical value of plasma protein Z detection.Methods Eighty-five patients who undertaken coronary arteriography were selected,and the patients were divided into coronary heart disease group (63 patients) and control group (22 patients)according to coronary arteriography results.The patients in coronary heart disease group were divided into three groups according to the Gensini score:A group (≤30 scores),B group (31-60 scores) and C group (＞ 60 scores).All patients' plasma was collected and stored at-80 ℃ until examined,and the plasma PZ level was detected by enzyme-linked immunosorbent assay method.Results The plasma protein Z level in coronary heart disease group was significantly lower than that in control group [(721.82 ± 289.53) μ g/L vs.(1 077.80 ± 338.12) μ g/L],and there was statistical difference (P＜ 0.05).The plasma protein Z level in A group,B group and C group was (856.09 ± 312.53),(665.27 ± 267.15) and (643.04 ±248.39) μg/L,respectively.The plasma protein Z level in B group and C group was significantly lower than that in A group,and there was statistical difference (P ＜ 0.05),but there was not statistical difference between C group and B group (P ＞ 0.05).There was negative correlation between the plasma Z level and Gensini score (r =-0.300,P =0.017).In coronary heart disease group,the plasma Z level in patients with smoking was significantly lower than that in patients without smoking [(687.83 ± 249.94) μ g/L vs.(844.29 ± 454.71) μ g/L,and there was statistical difference (P ＜ 0.05).There was negative correlation between the plasma Z level,age and hypersensitive C reactive protein (r =-0.349,-0.339,P ＜ 0.05).Conclusions Plasma protein Z level in patients with coronary heart disease is significantly decreased,and the plasma protein Z level has negative correlation with the severity of coronary atherosclerosis.Smoking can induce the decrease of plasma protein Z level,and the decrease of protein Z level maybe a predictor for coronary heart disease.

Objective To screen altered proteins of hippocampus in the stress-induced depression (STRID) rat model, and explore the potential molecular mechanism. Methods Twenty Sprague-Dawley rats were randomly divided into the control group and STRID group, 10 rats in each group. Chronic unpredictable mild stress (CUMS) methods including fasting for solids and liquids, electric foot-shock, reversing day and night, cold water swimming, cage tilt, scare stimulation and tail pinch were conducted on STRID rats with no repeats for 28 days to make up the depression animal model. The control group was normally fed during this period. After the stress stimulation, the hippocampus protein samples were used for two dimensional electrophoresis to screen the differentially expressed protein, and then mass spectrum identification and function analyze were conducted. Results Compared with the control group, 34 proteins were altered in STRID group. Among which, 18 were up-regulated, and 16 were down-regulated. The differentially expressed proteins mainly located in cytoplasm, mitochondrion, extracellular exosome and myelin sheath. The involved signaling pathways included metabolic pathway, oxidative phosphorylation pathway, and Alzheimer's disease, Parkinson's disease and Huntington's disease pathways. Conclusion The altered proteins and dysfunction of nerve signaling, and the excess of oxidative phosphorylation in hippocampus of STRID rats may be one of the pathogenesises.

This study investigated the abnormal expression of ATP synthase β-subunit (ATPsyn-β) in pancreas islets of rat model of polycystic ovary syndrome (PCOS) with type 2 diabetes mellitus (T2DM),and the secretion function changes after up-regulation of ATP5b.Sixty female SD rats were divided into three groups randomly and equally.The rat model of PCOS with T2DM was established by free access to the high-carbohydrate/high-fat diet,subcutaneous injections of DHEA,and a single injection of streptozotocin.The pancreas was removed for the detection of the ATPsyn-β expression by immunohistochemical staining,Western blotting and reverse transcription-PCR (RT-PCR).The pancreas islets of the rats were cultured,isolated with collagenase Ⅴ and purified by gradient centrifugation,and the insulin secretion after treatment with different glucose concentrations was tested.Lentivirus ATP5b was successfully constructed with the vector of GV208 and transfected into the pancreas islets for the over-expression of ATPsyn-β.The insulin secretion and intracellular ATP content were determined after transfection of the PCOS-T2DM pancreas islets with Lenti-ATP5b.The results showed that the expression of ATPsyn-β protein and mRNA was significantly decreased in the pancreas of PCOS-T2DM rats.The ATP content in the pancreas islets was greatly increased and the insulin secretion was improved after the up-regulation of ATPsyn-β in the pancreas islets transfected with lenti-ATP5b.These results indicated that for PCOS,the ATPsyn-β might be one of the key factors for the attack of T2DM.

Objective To investigate the role of three-dimensional scaffolds seeded with NeuroD1-modified neural stem cells (NSCs) for repair of spinal cord injury in rats.Methods A new three-dimensional bio-printer was used to make bionic spinal cord scaffolds.NSCs are transduced with retrovirus vectors encoding NeuroD1 to express transgenes in high levels.Forty healthy female SD rats were divided into control group,scaffold group,scaffold + NSCs-green fluorescent protein (GFP) group and scaffold + NSCs-NeuroD1 group with 10 rats per group,according to the random number table.Spinal cord injury in rats was induced using the electric controlled cortical impactor.A week later,the control group was excised 3 mm spinal cord at the injury site under microscope.RT-PCR was used to confirm the construction of NeuroD1 overexpressing NSCs.Survival and differentiation of transplanted NSCs were detected with fluorescent staining.Rat neurological motor function was evaluated with BBB score at postoperative 1,2,4,6 and 8 weeks.Rat electrophysiological changes were observed by monitoring motion evoked potential and sensory evoked potential at 8 weeks.Results RT-PCR results confirmed the successful reconstruction of NeuroD1-overexpressing NSCs.BBB score in scaffold + NSCs-NeuroD1 group was the highest and had significant differences compared to other three groups (P ＜ 0.05).Electrophysiological results showed the motor and sensory in scaffold + NSCs-NeuroD1 group had the shortest latencies and highest amplitudes,which revealed significant differences compared to other three groups (P ＜0.05).Immunofluorescence staining showed GFP cells in scaffold + NSCs-NeuroD1 group at 8 weeks,which differentiated into neurons and astrocytes.GAP-43 was positively stained,and myelin formation was detected.Conclusion Three-dimensional scaffolds seeded with NeuroD1-modified NSCs can promote nerve loop reconstruction in spinal cord injury rats,and accelerate recovery of motor and sensory function.

Objective To investigate the effects of synuclein-γ (SNCG) gene silencing on the proliferation and apoptosis of glioma U87-MG cells.Methods Five small hairpin RNA templates targeting SNCG and a negative control were synthesized and cloned into the lentiviral vector system and all the constructs were sequenced.Then the recombinant lentiviral vectors were used to infect U87-MG cells.The lentiviruses which can effectively inhibit protein expression levels of SNCG were selected by RT-PCR for further study.Colony formation and flow cytometry assay were used to investigate the effects of SNCG downregulation by RNA interference on the clony formation,proliferation,and apoptosis of U87-MG cells,respectively.Results The lentiviral vectors carrying 5 shRNAs targeting the SNCG gene were successfully constructed,and SNCG siRNA3 and siRNA5 showed higher interfering efficiency than other vectors.In comparison with the group of negative control,SNCG siRNA3 and siRNA5 were observed to significantly inhibit SNCG expression at the mRNA levels (the relative mRNA levels:siRNA3 (0.17± 0.01)％,siRNA5 (0.13±0.01)％ vs (1.00±0.10)％,P＜0.05).Also,SNCG suppression mediated by RNAi significantly inhibited the clone formation (colony number:siRNA3 (66± 12),siRNA5 (1 ± 1) vs (80± 5),P＜0.05),and the proliferation (ratio of cells in S phase:siRNA3 (41.2±0.7) ％,siRNA5 (39.9±0.5) ％ vs (47.6±2.2) ％,P ＜0.05),but promoted the apoptosis (cell apoptosis:siRNA3 (22.9± 0.4) ％,siRNA5 (28.6± 0.9) ％ vs (1.1 ± 0.1) ％,P＜0.01) of transfected U87-MG cells.Conclusion SNCG suppression at the mRNA level mediated by RNAi can inhibit the proliferation and the clony formation,but induce the apoptosis of glioma U87-MG cells in vitro,suggesting that SNCG suppression mediated by an RNAi strategy may become a novel approach for treating human gliomas.