Objective To investigate the effects of mild hypothermia ( MH ) on the expression of my- eloperoxidase(MPO) and cyclooxygenase 2 (COX 2) in rats after cerebral ischemia and reperfusion (CIR). Methods Forty-eight Wistar rats were randomly divided into four control groups (n=6 in each) and four MH groups (n=6 in each).CIR models were established by suture occlusion of the left middle cerebral artery.The rats in the MH groups,but not in the control groups,were treated with MH.Rats were killed at 4 h,8 h,12 h and 16 h after CIR.MPO expression was measured,along with the expression of COX 2 as measured by Western blot- ting and immunohistochemical methods.Results Compared with the control groups,MPO activity and the COX 2 expression in the cortex and striatum were significantly lower in all the MH groups at 4 h,8 h,12 h and 16 h after CIR.Conclusion MH treatment can protect neurons by decreasing MPO activity and COX 2 expression,allevia- ting inflammation and reducing secondary injuries after CIR.

Objective To explore the efficacy and safety of bedside continuous blood purification (CBP) in the treatment of neonatal multiple organ failure (MOF).Methods Totally 6 newborn infants of MOF were hospitalized in department of neonatology in our hospital from June 2011 to June 2013.These 6 cases of clinical data were retrospectively analyzed,6 neonates were treated with CBP combined with conventional treatment.The model for CBP was continuous veno-venous hemodialysis filtration (CVVHDF),blood flow velocity was 3 to 5 ml/(kg· min),replacement fluid dose was 20 to 30 ml/(kg· h),dialysis fluid dose was 15 to 25 rnl/(min· m2).The clinical outcome measures included,blood pressure,blood pH,K+,Na+,blood urea nitrogen,creatinine,urine volume,PaO2/FiO2 and epinephrine intravenous dose,respectively before CBP treatment,6 h,12 h,24 h,48 h after CBP treatment and the end of CBP treatment.The efficacy of CBP treatment was evaluated in neonatal MOF.Results Gestational age of 6 neonates with MOF was 33 to 41 weeks,2 to 19 days old,2.25 to 3.36 kg birth weight.Primary disease was 4 cases of neonatal septicemia(1 case with congenital hereditary metabolic disease),2 cases of severe neonatal asphyxia.All 6 cases of venous catheter were smoothly done.CBP treatment persisted for 49 to 106 hours.Compared with before CVVHDF treatment,blood K+,blood urea nitrogen,creatinine significantly decreased at 12 h after CVVHDF treatment [(5.32 ± 1.84) mmol/L vs.(9.81 ±3.61) mmol/L,(9.0 ±3.4) mmol/L vs.(12.8 ±6.1) mmol/L,(99 ± 16) μmol/L vs.(176 ±25) μmol/L,P ＜0.05],and reached the normal range at 24 h after treatment,urine volume significantly increased at 24 h after treatment (P ＜ 0.05).PaO2/FiO2 reached 200 mmHg (1 mmHg =0.133 kPa) at 6 h after treatment and more than 300 mmHg at 24 h after treatment(P ＜0.05).Fifty percent of epinephrine intravenous dose were down-regulation at 12 h after treatment and stopped using epinephrine at 48 h after treatment.CBP treatment of 6 cases showed effective.Conclusion Application of bedside CBP treatment in neonatal MOF is safe,can effectively help neonates with MOF to skip over renal failure stage.

In this paper,the significance of developing undergraduates’ creative qualities and how to train undergraduates’ innovative qualities are discussed.The effects of teaching reformation are also analyzed.

AlM: To investigate the effect of endocapsular phacoemulsification cataract extraction and intraocular lens (lOL) implantation with a 1. 8mm or 3. 0mm clear corneal incision on total root mean square ( RMS ) value of the cornea, corneal astigmatism, spherical aberration, coma, trefoil and tear film.
METHODS:ln a prospective study, 156 age- related patients ( 196 eyes ) were randomly distributed into two groups. 1. 8mm-group comprised 94 eyes that had a silicone lOL inserted through a 1. 8mm sutureless clear corneal incision, while, 3. 0mm- group comprised 102 eyes through a 3. 0mm clear corneal incision. Postoperatively, the changes in the total RMS value of the cornea, corneal astigmatism, spherical aberration, coma, trefoil and tear film at 1wk, 1 and 3mo were determined respectively.
RESULTS:ln both groups, postoperatively at 1wk,there were statistically significant differences ( P<0. 05 ) in the total RMS value of the cornea, corneal astigmatism, spherical aberration, coma, trefoil and tear film, while, there were statistically minimal differences ( P< 0. 05 ) between 1. 8mm-group and 3. 0mm-group at 1mo, but were not statistically significantly different ( P > 0. 05 ) between two groups at 3mo postoperative.
CONCLUSlON:This study confirms that incision size has strong impact on the corneal higher-order aberrations, especially, 3. 0mm incision caused significant differences in the total RMS value of cornea, corneal astigmatism, spherical aberration, coma, trefoil and tear film compared with 1. 8mm micro-incision, therefore, micro-incision is very beneficial for clinical use in phacoemulsification.

Objective To detect the impact of reactive oxygen species ( ROS) on mitochondrial morphology and distri-bution during mitosis.Methods A viral vector in which the fluorescence gene was specifically under the control of mito-chondrial promoter was constructed and confirmed through DNA sequencing and Western blotting.After transfecting HeLa s3 cell with packaged virus, the HeLa s3-COX4tp-EGFP cell line stably expressing the mitochondrial fluorescence signal was obtained.With immunofluorescent staining, the impact of ROS on the morphology and distribution of mitochondria dur-ing mitosis was inspected.Result The cell line constantly expressing mitochondrial fluorescence signals was successfully constructed.Meanwhile,it was found that H2 O2 treatment could significantly change the morphology and distribution of mi-tochondria during mitosis by confocal microscopy.Conclusion Our study demonstrates that ROS can affect the morphology and distribution of mitochondria during mitosis.This research help study the relationship between the mitochondrial function and the regulation of mitosis in the future.

Objective To explore the efficacy and safety of mild hypothermia (MH) in treating the infants with moderate-to-severe neonatal hypoxic-ischemic encephalopathy(HIE),and to make a follow-up of the nerve motor development of the infants at 18 months old after discharge.Methods Totally 61 neonates with moderate-to-severe HIE in Neonatal Intensive Care Unit (NICU) from Jan.2007 to Dec.2013 were retrospectively analyzed.According to before and after MH therapeutic apparatus was used by NICU of Shanghai Children's Hospital,61 neonates of HIE were divided into 2 groups,the conventional treatment group(25 cases) and MH treatment group(36 cases).The patients in both groups were measured respectively by using the amplitude integrated electroencephalography (aEEG) before MH treatment and at 72 hours after M H treatment,by neonatal behavioral neurological assessment(NBNA) on the 28th day after birth,and by adopting Bayley Scales of Infant Development at 18 months old.The adverse reactions,serious disability cases and deaths of MH treatment were recorded.Results Compared with the conventional treatment group,aEEG recording before treatment showed no statistically significant differences in MH treatment group [maximum voltage:(22.4 ±3.1) μV vs(18.6 ±2.5) μV,maximum voltage:(8.2 ±2.6)μV vs(6.5 ±1.9) μV,t =1.264,0.852,all P ＞ 0.05].However,aEEG recording at 72 h after treatment showed statistically significant differences in MH treatment group [maximum voltage:(24.1 ± 3.2) μV vs (30.6 ± 2.8) μV,maximum voltage:(9.7 ± 3.4) μV vs (13.3 ± 2.2) μV,t =6.376,4.257,all P ＜ 0.05].Severe disability cases [24.0％ (6/25 cases) vs 5.6％ (2/36 cases),x2 =4.405,P ＜ 0.05] and deaths [16.0％ (4/25 cases) vs 0 (0/36 case),x2 =6.1 64,P ＜ 0.05] in MH treatment group were significantly decreased,and there was significantly difference in NBNA on the 28th day after birth[(35.9 ± 2.1) vs(39.1-± 1.6),t =3.361,P ＜ 0.05],and scales of neurobehavioral evaluation through follow-up of 18 months old [mental development index (MDI):(85.2 ± 10.7) vs (96.5-± 13.1),t =7.839,P ＜ 0.05].Very few neonates had apnea,coagulation dysfunction,arrhythmia and other adverse reactions in MH treatment course.Conclusions MH treating moderate-to-severe HIE is safe and effective.MH is effective in reducing death and major disabilities in neonates with moderate-to-severe HIE and without significant side effects.MH can obviously improve the development of nervous system disorders in 0-18 months infants,and can significantly improve these infants' Bayley developmental scale neurobehavioral scores.

Objective To explore the clinical features, diagnosis, and treatment of non-immunologic hydrops fetalis (NIHF). Methods The clinical data of 10 cases of NIHF in neonatal intensive case unit during January 2011 to December 2016 were analyzed retrospectively. The related literatures were reviewed. Results In 10 cases of NIHF (6 males and 4 females). the gestational age were 32-42 weeks, and the birth weight was 2.25-3.95 kg. Among them, there were 3 cases of infectious diseases (cytomegalovirus, Streptococcus agalactiae, and parvovirus infection, one case each), 2 cases of fetal cardiovascular abnormalities, 2 cases of chromosomal abnormalities, 1 case of abnormal thoracic structures, 1 case of twin transfusion syndrome, and 1 case of etiology unknown of fetal hydrops. The clinical manifestations showed that there were 8 cases with 2 or more areas of edema (or hydrops), and only 2 cases with skin edema. Finally, 6 cases were cured and discharged, 2 cases were discharged by themself, and 2 cases died. Conclusions Prenatal ultrasound is a reliable method for the diagnosis of NIHF. Fetal edema in early pregnancy, especially with congenital malformations, is recommended for termination of pregnancy. After birth, NIHF should be diagnosed promptly so as to avoid or reduce severe complications.

AIM: To analyze the subcellular localization of Arnt2 in rat cerebellar granule neurons (CGNs). METHODS: Based on the amino acids sequence of Arnt2 (LOCUS:NP_036913), the subcellular localization of Arnt2 in eukaryotic cells and the nuclear export signals (NES) of Arnt2 were predicted in CBS bioinformatics database. The subcellular localization of Arnt2 in rat cerebellar granule neurons was detected by the method of laser scanning confocal microscopy (LSM) analysis. RESULTS: It was predicted that Arnt2 located in nuclei of eukaryotic cells with the most probability, while located in cytoplasmic mitochondria with a slight possibility. A nuclear export signal was found in Arnt2 amino acids sequence, it was identified to be the leucine of No.143 that located in N-terminal of Arnt2 amino acids sequence. Finally, the result of LSM analysis shows nuclear localization of Arnt2 in rat CGNs. CONCLUSION: Arnt2 is located in nuclei of normal rat CGNs, it suggests that Arnt2 has the tendency to translocate into mitochondria after induced by some of inducible factors, for both the possibility of mitochondria localization and NES exist in Arnt2 amino acids sequence.

AIM: To observe the expression of neuronal Aryl hydrocarbon receptor nuclear translocator 2 (ARNT2) involved in neuronal apoptosis evoked by low K + and to investigate the relationship between ARNT2 and neuronal apoptosis. METHODS: After neuron apoptosis model was established, the changes of mRNA and protein of ARNT2 during apoptosis were investigated by RT-PCR and Western blotting, respectively. Immunofluorescence was analyzed by confocal microscopy to probe the subcellular localization of ARNT2. RESULTS: Induced by low K +, the expression of ARNT2 mRNA was up-regulated obviously at the point of 30 min, and peaks at the point of 1 h. This up-regulated expression lasted for 12 h, and the variation of ARNT2 protein was similar to that of mRNA. The results of immunofluorescence analyzed by confocal microscopy showed that the localization of ARNT2 protein was in the nucleus. CONCLUSION: ARNT2 locate in nuclei of normal cerebellar granule neurons of rat. During the process of apoptosis evoked by low K +, both mRNA and protein of ARNT2 are overexpressed.

Objective:The immunological effects of HCV-DNA vaccine with different adjuvants were detected by ELISPOT in mice.Methods:Female BALB/c mice were primed with naked HCV-DNA, HCV-DNA encapsulated by liposome DDAB/EPC or DC-Chol/DOPE, HCV-DNA mixed with Montanide ISA 720 or aluminum hydroxide, respectively, and boosted twice accordingly in a four-week interval. Cytokine production by splenocytes was assessed by ELISPOT.Results:In most cases, splenocytes from mice vaccinated with DDAB/EPC liposome produced more IFN-?. These splenocytes also have significant higher IL-2 production compared with the other groups. In expansion with NS5b, splenocytes from alum group have significance in IL-4 production compared with other groups. The profile of cytokine production revealed that the INF-? overwhelmed IL-4 in naked DNA, DDAB/EPC, and DC-Chol/DOPE groups while IL-4 surmounted IFN-? in alum and Montanide groups.Conclusion:Encapsulation with liposome DDAB/EPC has the strongest adjuvant effect in inducing Th1 dominated immunity. Alum and Montanide can convert the Th1 nature of DNA vaccine to Th2-biased immunity.