OBJECTIVETo explore the effect of 3-n-butylphthalide pretreatment on the delayed neuronal death(DND) and the expreesion of heat shock protein70 (HSP70) in rat hippocampus after ischemia/ reperfusion.

METHODSAll rats were randomly divided into sham group (n = 36), total cerebral ischemia (TCI) group (n = 36), butylphthalide (NBP) group (n = 6), NBP + TCI group( n = 36), quercetin + NBP + TCI group (n = 6), dimethyl sulfoxide (DMSO) + NBP + TCI group (n = 6). The model of total cerebral ischemia/reperfusion was established by blocking vertebral arteries and carotid arteries. In sham group, TCI group and NBP group, the animals were further divided into instantly, 6 h, 12 h, 1 d, 3 d, 5 d groups according to the time interval after sham operation or TCI. Histological changes of the hippocampus were evaluated using thionin staining under light microscope by determining the delayed neuronal death (DND) and the expression of HSP70 was assayed using immunohistochemistry.

RESULTSNBP pretreatment could reduce delayed neuronal death in CA1 of hippocampus induced by TCI-reperfusion injury in rats, and up-regulated the expression of HSP70 in CA1 hippocampus of brain ischemic/reperfusion for 5 days. Quercetin blocked the acquirement of the brain ischemic tolerance induced by NBP preconditioning.

CONCLUSION3-n-butylphthalide (NBP) prevents the neurons from ischemia/reperfusion injury through upregulating the expression of HSP70.

Animals ; Benzofurans ; pharmacology ; CA1 Region, Hippocampal ; cytology ; pathology ; Cell Death ; Cerebral Infarction ; drug therapy ; HSP70 Heat-Shock Proteins ; metabolism ; Ischemic Preconditioning ; Neurons ; cytology ; Rats ; Rats, Wistar ; Reperfusion Injury ; drug therapy

BACKGROUND:Bone marrow mesenchymal stem cells(MSCs)are a kind of adult stem cells with multi-potential differentiation property.At present,it has served as cell carrier for the treatment of Parkinson's disease.OBJECTIVE:To construct pDsRed-C1-CDNF eukaryotic expression vector and induce its expression in rat MSCs.METHODS:CDNF gene was amplified from mouse tissues using RT-PCR,and sequence with Xho I,BamHI restriction enzyme cutting site.The CDNF gene was inserted into the eukaryotic expression vector pDsRed-C1 encoding red fluorescent protein gene.The plasmid pDsRed-C1-CDNF was constructed and transfected into rat bone marrow MSCs.RESULES AND CONCLUSION:The pDsRed-C1-CDNF recombinant plasmid was confirmed by double digestion of Xho I and BamHI restriction enzyme or single digestion of BamHI,and PCR sequence.Results show that the recombinant pDsRed-C1-CDNF eukaryotic expression vector was successfully constructed.

Objective To establish the quality standards of Zizhu ointment. Methods The TLC was applied to identify Radix arnebiae and borneol of Zizhu ointment, and the content of borneol was determined by gas chromatography. Results The TLC spots were clear, well-separated and easy to identify. The good linear range of borneol reference substance on calibration curve was 0.048 4-1.210 0 μg, and the recovery was 90%-110%, the relative standard deviation was less than 5%. Conclusion The method is simple and feasible, can be used as the quality control method of Zizhu ointment.

Objective To investigate the expression of angiotensinⅡ (Ang Ⅱ ) and its type 1 receptor (AT1) in circulation, placenta and kidney of the rots preeclampsia. Methods Preedampsia rat model was developed by inhibitor of nitric oxide synthase (L-NAME). The systolic blood pressure (SBP), 24 h urine protein, hepatic and renal function were compared among the precelampsia group, the normal pregnant group and nonpregnant control group. The kidney tissue was observed by light microscopy. ELISA and radioimmunoassay were used to detect Ang Ⅱ in rat plasma and kidney homogenate respectively. Placental AT1 was measured by Westem blot. The level of kidney AT1 was evaluated by immunohistochemistry and Western blot. Results In preeclampsia rats, SBP and 24 h urine protein were significantly higher compared with control groups. Compared to normal pregnant group, plasma Ang Ⅱ of preeclampsia rats was much higher [(0.706±0.086) ng/L vs (0.540±0.085) ng/L, P＜0.05]; placental AT1 was increased by 46%(P＜ 0.05); kidney Ang Ⅱ was decreased signigicantly [(65.543±40.634) ng/g vs (165.543±33.078) ng/g, P＜0.05]. The expression of ATI in kidney of preeclampsia rats was reduced evidently,which was only 33% of normal pregnancy group and 59% of nonpregnant control greup,respeetively (P＜0.05). Conclusions In preeclampsia rat model, the circulating Ang Ⅱ is increased, the placental RAS isactivated, while the kidney BAS is suppressed. The underlying mechanism of proteinuria and kidney damage associated with this phenomenon in preeclampsia needs further research.

Objectives: To evaluate the clinical application of emergency extracorporeal circulation(ECC) in cardiac surgery. Methods: Emergency open heart surgery was performed in 24 ECC cases. Warm blood left heart bypass was performed in 3, hypothermic cardiopulmonary bypass in 21. The vena cava-aorta extracorporeal circulation was built in 16,vena cava-femoral in 4, left atrium-femoral in 3 and femoro-femoral in 1. Retrograde or antegrade perfusion through aortic root were applied to protect the heart. Results: There were 2 hospital deaths, the hospital mortality rate was 8.3%. The cardiopulmonary bypass was established in 10 to 15 minutes with an average duration of cardiopulmonary bypass of 78～132 minutes. Twenty-two patients were discharged from the hospital and have resumed a normal and active life style. Conclusions: Emergency cardiopulmonary bypass support is a powerful resuscitative tool that may stabilize the condition of patients with cardiogenic shock and mutiorgan dysfunction to allow for definitive intervention. The femoro-femoral cannulation is the first choice for emergency. The time to establish extracorporeal circulation should be as short as possible.

Objective To observe levosimendan-induced hemodynamic improvement of chronic decompensated heart failure patients and adverse effects,and sum up the nursing experience of Levosimendan.Methods Sixty-three patients with decompensated heart failure were treated with Levosimendan,based on optimized conventional treatment for heart failure.Aminoterminal pro-brain natriuretic peptide (NT-proBNP) and left ventricle ejective fraction (LVEF) were measured at baseline and one week after Levosimendan administration.In addition,we summarized the clinical observation,adverse effects and nursing experiences of treatment with Levosimendan in decompensated heart failure patients.Results Levosimendan increased 24-hour urine output in patients at the first day after treatment.This effect of levosimendan persisted for 7 days at least.We observed an improvement in LVEF and a reduction of NT-pro-BNP at 7 days after the initiation of levosimendan treatment.Clinical efficiency of levosimendan in patients was 84.1％ in this study.Twenty two patients(34.9％) experienced different adverse events.Conclusions Levosimendan therapy can improve cardiac function and reduce BNP in patients with decompensated heart failure.The rate of adverse effects of Levosimendan is high.It is important to enhance therapy effect by careful nursing and observing variety state during treatment with Levosimendan.These data will be useful for the nursing care of Levosimendan in clinical practice.

Objective:To explore the effect of multi-channel health education model by professional team on drug compliance, disease knowledge and recurrence in rate of inflammatory bowel disease (IBD) patients, so as to provide basis for formulating scientific health education measures.Methods:From February 2016 to February 2019, IBD patients visited First Affiliated Hospital of Kunming Medical University were selected. According to whether they received health education, the patients were divided into intervention group (100 cases) and control group (138 cases). Morisky medication adherence scale-8 (MMAS-8) and Chinese version of Crohn′s and colitis knowledge score (CCKNOW) were used to evaluate treatment compliance and disease knowledge. The score of MMAS-8, the proportion of poor drug compliance, CCKNOW score and recurrence rate at 48 weeks of follow-up were compared between the intervention group and control group. Two sample t test and chi-square test were used for statistical analysis. Results:The total scores of MMAS-8 and CCKNOW of the intervention group were both higher than those of the control group (5.58±1.96 vs. 4.47±1.44, 10.87±4.21 vs. 9.23±4.65), and the differences were statistically significant ( t=-5.06 and -2.79, both P<0.05). The proportion of patients with poor drug compliance and recurrence rate at 48 weeks of follow up of the intervention group were both lower than those of the control group (56.0%, 56/100 vs. 86.2%, 119/138; 20.0%, 20/100 vs. 31.9%, 44/138), and the differences were statistically significant ( χ2=38.18 and 4.17, both P<0.05). Conclusions:Multi-channel health education by professional team can effectively improve the drug compliance and disease knowledge in IBD patients, improve patient self-management ability, and reduce the recurrence rate.

Background The human transforming growth factor beta-induced gene (TGFBI) is the first determined pathogenic gene to corneal dystrophy.But the molecular genetic mechanism is completely unknown.The study of concerning role of TGFBI is very important for us understand the physiological function of cornea,and the pathogenesis of corneal dystrophy.Objective The vector of human transforming growth factor beta-induced gene (TGFBI) in eukaryotic expression was constructed and transfected into the human corneal epithelial cells in order to explore its influence on the growth of human corneal epithelial cells.Methods Total RNA was extracted from normal donor cornea tissue and cDNA was obtained by reverse transcription.TGFBI cDNA was synthesized by reverse transcription-PCR and cloned into pCMV-N-HA vector and identified by sequencing with PCR and EcoRV,XhoI double restriction endonuclease.The cells were grouped into recombinant pCMV-N-HA-TGFBI plasmid group,pCMVN-HA plasmid group,non-transfected group and pGFP-C2 transfected group.The recombinant pCMV-N-HA-TGFBI plasmid was transfected to human corneal epithelial cells and identified by observing the expression of enhanced green fluorescence protein(EGFP) in the cells.The TGFBI mRNA and proteins were harvested from the cells for real-time PCR analysis and Western blot assay respectively in 58 hours after transfection.The growth of the transfected cells was assessed by Cell Counting Kit-8.The expressions of matrix metalloproteinase(MMP) and tissue inhibitors of matrix metalloproteinase (TIMP) proteins and their mRNA in transfected cells were detected using SYBR fluorescence realtime PCR analysis and Western blot assay.Results The sequencing result of pCMV-N-HA-TGFBI positive clone plasmid showed that amplified TGFBI eDNA inserted into the vector at the correct sequence.EGFP was expressed in transfected cells in 48 hours after transfer of pGFP-C2 with the transfer efficacy 70％.The expression intensity of TGFBI mRNA was significantly higher in recombinant pCMV-N-HA-TGFBI plasmid group compared with pCMV-N-HA plasmid group and non-transfected group,and TGFBI protein was expressed in recombinant pCMV-N-HA-TGFBI plasmid group.No significant difference was found in the A450value among recombinant pCMV-N-HA-TGFBI plasmid group,pCMV-N-HA plasmid group and non-transfected group ( F=3.34,P＞0.05 ).The mRNA level of MMP1,MMP3in the transfected cells was significant elevated but that of TIMP1 was declined in the recombinant pCMV-N-HA-TGFBI plasmid group compared with pCMV-N-HA plasmid group and non-transfected group (all P ＜ 0.05 ).Meanwhile,the expressions of MMP1,MMP3 and TIMP1 proteins appeared the same tendency( all P＜0.05).Conclusions Eukaryotic expression vector harboring human TGFBI eDNA can be successfully constructed and efficiently overexpressed in human corneal epithelial cells.TGFBI gene is involved in the physical and pathological conditions of human corneal epithelial cells by regulating the activity of MMP1,MMP3 and TIMP1.The results offer a new approach for the study of the role of TGFBI in pathogenesis of corneal transparency.

We wished to select a cold-adapted genotype G1P[8] ZTR-68 rotavirus (China southwest strain) in MA104 cells for possible use as a live vaccine. ZTR-68 was recovered originally from children with diarrhea. The virus was cultivated at 37 degrees C at the first passage. Then, the cultivation temperature was decreased stepwise by 3 degrees C per eight passages. In total, the virus was passaged 32 times, and cultivation was terminated at 28 degrees C. Biological characteristics of the virus were analyzed during serial passages. There was no difference between the migration patterns of genomic dsRNA segments according to polyacrylamide gel electrophoresis of original and cold-adapted viruses. Infectious and red cell-agglutination titers of cold-adapted virus were lower than those of the parent virus. Also, the virus formed small-size plaques with irregular shapes at 31 degrees C and 28 degrees C. These results suggested that a genetically stable attenuated virus can be obtained through serial cold-adapted passages. Thus, an alternative strategy is provided by cold-adaption for development of attenuated live rotavirus vaccines.
Adaptation, Physiological ; China ; Cold Temperature ; Diarrhea ; virology ; Female ; Genotype ; Humans ; Infant ; Male ; Rotavirus ; genetics ; growth & development ; isolation & purification ; physiology ; Serial Passage ; Virus Cultivation ; Virus Replication

Objective To investigate the possible correlation of the clinical parameters, such as age, obesity, Epworth sleepiness score (ESS), with the severity of snoring and obstructive sleep apnea hypopnea syndrome (OSAHS). Methods One hundred and forty-eight patients with snoring during sleep admitted from May to Jul. 2008 were asked to answer the questions from a questionnaire concerning snoring, daytime sleepiness, and habits such as smoking and drinking, etc. All patients underwent at least a polysomnography (PSG) and the physical examination included height, weight, and body mass index (BMI). Age, BMI, the lowest SaO_2(%), ESS score, the biggest reduction of oxygen (%), a total suspension of time, the average correlation between respiratory disorder index (RDI) applied computing Pearson correlation test. Simple snoring and OSAHS group of mild, moderate and severe inter-group comparison analysis using generalized linear models. Results The prevalence of OSAHS was increased with age, higher in males than in females. A statistically significant correlation between ESS, BMI, the lowest SaO_2 with the RDI was detected. The difference of ESS, the lowest SaO_2 and the BMI was significant between the different serious patients (P<0.05). Conclusions OSAHS has a high morbidity rate in outpatients with snoring. Age and obesity are liability factors of OSAHS. BMI, the lowest SaO_2, ESS and RDI have well correlationship, which can be used to assess the pathogenetic condition, even make a primary diagnosis.