Purpose To construct a lentiviral ventor-mediated RNA interference of human midkine ( MK) gene and to provide the ba-sis for further experiment in vivo and in vitro. Methods Four RNAi sequences targeting human MK gene were designed, and cloned into the lentiviral vector to construct lentiviral vectors:GV115-MK-1, GV115-MK-2, GV115-MK-3 and GV115-MK-4. After transfec-tion into competent E. Coli bacteria, the candidate clones were identified by PCR and DNA sequencing. The titer of lentiviruses was determined after 293 T cells were co-transfected with GV115-MK, pHelper 1. 0 and pHelper 2. 0. The four kinds of recombinant lenti-viruses were used to infect human breast cancer cell MDA-MB-231, and the expression levels of MK mRNA were detected by real-time PCR. Results PCR analysis and DNA sequencing confirmed that the four inserted MK RNAi sequences were corrected. Strong green fluorescence was observed in the 293 T cells under the fluorescent microscope after co-transfection. The titer of the concentrated virus was 6 × 108 , 5 × 108 , 5 × 108 and 6 × 108 TU/ml, respectively. The MK expression in MDA-MB-231 cells infected with lentiviral vec-tors GV115-MK-1 was decreased by 87. 2% (P<0. 05). Conclusion Lentiviral RNAi vector for MK gene is successfully construc-ted, and it could effectively inhibit the expression of MK gene in MDA-MB-231 cells in vitro.

Objective To compare three methods for detecting extend-spectrum ?-latamases (ESBL s) and investigate current resistance of ESBL s.Methods 538 isolates of Enterobacter were detected by VITEK-AMS, compound piece type confirm test, and double-disk synergy test.Results 20.1% (108/538) was found to be ESBL s positive by VITCK-AMS. The positive rate of ceftaxime/clavulanic acid and ceftaxime compound pieces test was 19.5% (105/538). The positive rate of double-disk synergy test was 13.0%. The resistant rate of 12 antibiotics to ESBL s positive strains was significantly higher than ESBL s negative strains.Conclusions It is important to select the proper and rapid method to detect ESBL s in time.

Objective To investigate the effect of sufentanil and midazolam assisted epidural anesthesia for cholecystectomy on sedative effects,visceral pulling reaction and the functions of respiratory and circulatory system.MethodsFourty patients underwent cholecystectomy received informed consent, were randomly allocated to two groups:group A and group B( n =20 each). When the level of epidural block is appropriate,auxiliary medicine infusion was given 5min before the skin incision. In group A,first injection was sufentanil 0. lμg · kg-1 and midazolam 0. 02mg · kg-1 in 1-2 min, then continuous infusion of sufentanil and midazolam was given at the rate of 0. 2μg ·kg-1 · h-1 and 0. 04μg · kg-1 · h-1 respectively,and the infusion was stopped at the time of wound closure. Group B received slow intravenous injection of pethidine 1mg · kg-1 and droperidol 0. 5mg · kg-1. We recorded the differences of SBP、DBP、SpO2 、HR、Ransay score and visceral pulling reaction classification. Results SBP、DBP、HR after anesthesis,were decreased in different degree in the above groups,but there was no statistically significant difference( P ＞0. 05). According to Ramsay score,in group A there were 3 cases in grade Ⅱ、8 cases in Ⅲ and 9 cases in Ⅳ ,while in group B there were respectively 4 cases in Ⅱ、13 cases in Ⅲ and 3 cases inⅣ. By comparision,the differences had statistical significance ( u = 3.75, P ＜ 0. 05 ); Group A according to visceral pulling reaction classification (0- Ⅲ ), were 9 cases in grade 0,9 cases in Ⅰ , 1 cases in Ⅱ and 1 cases in Ⅲ, while in group B there were respectively 4,5,7 and 4 cases. By comparision, the differences had statistical significance(u = 4. 01,P ＜ 0. 01 ). Conclusion In the cholecystectomy,sufentanil and midazolam assisted epidural anesthesia,could improve the level of sedation ,prevent visceral pulling reaction ,and had a minor interference to respiratory and circulatory function.

Objective To investigate the signaling role of arachidonic acid in the invasion of RAW264.7 macrophage by Toxoplasma gondii . Methods Rate of infection was calculated by both light microscope and flow cytometer. Fluorescent emission spectra were recorded using a microspectrofluometer for the concentration of cytoplasmic free calcium. Results Calcium concentration in macrophages and rate of infection increased with a higher concentration of exogenous arachidonic acid in a dose-dependent manner. The invasion was dependent on the mobilization of calcium from the extracellular medium and from intracellular stores and followed the influx of calcium into the parasitized cell. Conclusion Arachidonic acid may enhance the rate of infection via calcium transduction pathway.

Objective To investigate the epidemiological and clinical characteristics and prognosis of 31 children with hand foot and mouth disease(HFMD) complicated with acute flaccid paralysis(AFP).Methods Thirty-one HFMD children complicated with AFP served as study objective,who were admitted to Tangshan Maternity and Children Health Hospital from Jun 2011 to Dec 2014.The clinical manifestations and follow-up outcomes of 31 children with HFMD followed by AFP were retrospectively reviewed.Results Participants in this study consisted of 31 children(16 males,15 females,age from 3 months to 9 years) who met the criteria for HFMD with AFP.Among the 31 cases,27(87.1％)cases were less than 3 years old and 29(93.5％) cases were from countryside.Eleven cases (35.5％)were infected by enterovirus 71,1 case (3.2％) was infected with coxsackieviruses A16 and 19 cases(61.3％) were infected by other enteroviruses.AFP developed(7.1 ±2.9) days after the onset of fever and progressed to maximum severity within 1-2 days.All of the cases presented with fever and skin rash,38.7％ cases presented with limbs tremor and 87.1％ cases presented with startle and skip.Besides AFP of limbs, 100％ cases complicated with encephalitis.Thirty-one cases showed poliomyelitis-like syndrome(20 cases with one flaccid limb,5 cases with lower limbs,4 cases with upper limbs and 2 cases with left hemiplegia).In these cases, the muscle power varied from level 0 to level 4.The muscle strengthen and muscle tone in 21 patients were recovered within 2 weeks,and the other cases showed recovery 2 to 3 weeks later.After four weeks, the muscle strengthen in 19 cases recovered to 5 level(10 cases with single lower limb,5 cases with single upper limb,3 cases with two lower limbs and 1 case with two upper limbs).Conclusion HFMD complicated with AFP most commonly occurs in children aged less than 3 years old.The majority of cases were from countryside.Enterovirus 71 is still relatively common pathogen.All the 31 cases complicated with encephalitis.Cases clinically presented one flaccid limb, lower limbs, upper limbs and hemiplegia.AFP may be to some degree reversible in HFMD cases.In most cases described here, paralysis occurred in a single lower extremity and recovered more rapidly than those with two fimbs affected or with single upper extremity impairment.

Objective To investigate the effect of midazolam combined with fentanyl as adjuvant therapy on inflammatory factors and biochemical indexes in children with serious hand-foot-mouth disease with mechanical ventilation .Methods One hundred and thirty children with serious hand-foot-mouth disease treated with mechanical ventilation were selected at Maternal and Child Health Care Hospital from January 2010 to January 2014 . The patients were divided into two groups according to the treatment regimen :58 cases treated with midazolam for sedation and analgesia as control group and 72 cases treated with midazolam combined with fentanyl as observation group . Inflammatory factors (interleukin-6 , interferon-γ ,high-sensitivity C-reactive protein and tumor necrosis factor-α) and biochemical indices (albumin ,alanine transaminase [ALT] ,aspartate transaminase [AST] ,alkaline phosphatase ,glutamate transpeptidase [γ-GT] and fasting blood glucose) before drug exposure and on withdrawal were compared between two groups .Adverse reactions were analyzed in the two groups .Continuous variables were compared using two-sample t-test , while categorical variables were compared using chi-square test . Results Interleukin-6 ,interferon-γ, high sensitive C reactive protein and tumor necrosis factor-α on withdrawal decreased significantly in both groups than those before drug exposure (all P< 0 .05) .All observation indices on drug withdrawal in observation group were significantly lower than control group (all P< 0 .05) .Levels of albumin ,ALT ,AST ,alkaline phosphatase and γ-GT were not significantly different between two time points in both groups (all P>0 .05) .However ,in observation group ,fasting blood glucose level decreased significantly on drug withdrawal compared with that before drug exposure ([5 .17 ± 0 .28] vs [10 .31 ± 1 .39] mmol/L ,t=46 .237 ,P=0 .000) ,and that was also lower than control group ([5 .17 ± 0 .28] vs [5 .85 ± 0 .34] mmol/L ,t=4 .372 ,P=0 .000) .Incidence of adverse reactions in observation group was significantly lower than control group (15 .3% vs 32 .8% ,χ2=4 .707 ,P=0 .030) . Conclusions Midazolam combined with fentanyl as adjuvant therapy is helpful to improve blood glucose , stabilize biochemical indices and reduce inflammation factor secretions in children with serious hand-foot-mouth disease with mechanical ventilation .This therapy is safe and worthy of clinical use .

Objective To explore the predictive value of contrast-enhanced echocardiography com-bined with serum levels of CD137 and insulin-like growth factor binding protein 6(IGFBP-6)for cardiovascular adverse events(MACE)in elderly patients with stable coronary heart disease(CHD)after percutaneous coronary intervention(PCI).Methods A total of 108 elderly patients with stable CHD(CHD group)who visited Department of Cardiology of Changsha First Hospital from March 2020 to March 2022 were recruited in this study.They were grouped into a non-MACE group(81 cases)and a MACE group(27 cases)according to whether MACE occurred after PCI.Another 100 healthy individuals who taking physical examination during the same period served as control group.Their serum CD137 and IGFBP-6 levels were detected,and the contrast agent filling speed(β value)and maximum number of microbubbles(A value)were calculated based on the results of contrast-enhanced echocardiography.Their general clinical data were col-lected.ROC curve analysis and multivariate logistic regression analysis were used to analyze the data.Results The serum levels of CD137 and IGFBP-6 were significantly higher,while the β value and A value were obviously lower in the CHD group than the control group(P＜0.01).And the serum levels were notably higher,and the β value and A value were remarkably lower in the MACE group than the non-MACE group(P＜0.01).The AUC of cardiac ultrasound parameters βvalue and A value combined with serum CD137 and IGFBP-6 to predict MACE after PCI in CHD patients was 0.930,which was significantly higher than the AUC value of every single indicator(P＜0.01).β value,A value,CD137 and IGFBP-6 levels were all risk factors for the occurrence of MACE in CHD patients after PCI(P＜0.01).Conclusion Contrast-enhanced echocardiography,serum CD137 and IGFBP-6 levels have certain predictive value for MACE in elderly CHD patients after PCI,and combined detection has higher predictive value.

Background/Aims: Cancer stem cells (CSCs) are believed to drive tumor development and metastasis. Activin and hepatocyte growth factor (HGF) are important cytokines with the ability to induce cancer stemness. However, the effect of activin and HGF combination treatment on CSCs is still unclear. Methods: In this study, we sequentially treated colorectal cancer cells with activin and HGF and examined CSC marker expression, self-renewal, tumorigenesis, and metastasis. The roles of forkhead box M1 (FOXM1) and sex-determining region Y-box 2 (SOX2), two stemness-related transcription factors, in activin/HGF-induced aggressive phenotype were explored. Results: Activin and HGF treatment increased the expression of CSC markers and enhanced sphere formation in colorectal cancer cells. The tumorigenic and metastatic capacities of colorectal cancer cells were enhanced upon activin and HGF treatment. Activin and HGF treatment preferentially promoted stemness and metastasis of CD133 + subpopulations sorted from colorectal cancer cells. FOXM1 was upregulated by activin and HGF treatment, and the knockdown of FOXM1 blocked activin/HGF-induced stemness, tumorigenesis, and metastasis of colorectal cancer cells.Similarly, SOX2 was silencing impaired sphere formation of activin/HGF-treated colorectal cancers. Overexpression of SOX2 rescued the stem cell-like phenotype in FOXM1-depleted colorectal cancer cells with activin and HGF treatment. Additionally, the inhibition of FOXM1 via thiostrepton suppressed activin/HGF-induced stemness, tumorigenesis and metastasis. Conclusions Sequential treatment with activin and HGF promotes colorectal cancer stemness and metastasis through activation of the FOXM1/SOX2 signaling. FOXM1 could be a potential target for the treatment of colorectal cancer metastasis.

Objective:To investigate the differences of white matter diffusion properties between vulnerable and resistant individuals to continuous attention after sleep deprivation.Methods:According to the psychomotor vigilance test performance before and after sleep deprivation, the participants were divided into the vulnerable group( n=24) and resistant group( n=25). All participants underwent diffusion tensor imaging (DTI) scans.Tract based spatial statistics(TBSS) was used to compare fractional anisotropy(FA), mean diffusivity(MD), axial diffusivity(AD), radial diffusivity(RD) maps between the two groups.Spearman correlation analysis was conducted by SPSS 24.0 to investigate the relationships between the altered DTI metrics and PVT task performance. Results:(1) Compared with resistant group, FA value of vulnerable group decreased in the body of corpus callosum(x, y, z=-8, 9, 25, t=-7.855), right superior longitudinal fasciculus(x, y, z=-39, -7, 26, t=-6.252), bilateral anterior limb of internal capsule(x, y, z=-13, 8, 13, t=-5.235; x, y, z=12, 8, 3, t=-5.024) and right posterior thalamic radiation(x, y, z=-26, -56, 17, t=-5.469)(TFCE corrected, P<0.05, cluster size≥50 voxel). (2) Compared with resistant group, MD value of vulnerable group increased in the body of corpus callosum(x, y, z=-3, -6, 26, t=7.613), right superior longitudinal fasciculus(x, y, z=-31, -19, 38, t=5.314), bilateral anterior limb of internal capsule(x, y, z=-16, 7, 8, t=6.898; x, y, z=15, 5, 7, t=6.652), splenium of corpus callosum(x, y, z=27, -53, 17, t=6.541), and AD value increased in the right superior longitudinal fasciculus(x, y, z=-33, -19, 39, t=4.892), splenium of corpus callosum(x, y, z=-22, -49, 21, t=5.450), genu of corpus callosum(x, y, z=4, 26, 0, t=4.332), as well as RD value increased in the right superior corona radiata(x, y, z=-17, 1, 33, t=7.558), body of corpus callosum(x, y, z=4, -8, 26, t=6.699), right anterior limb of internal capsule(x, y, z=-12, 7, 3, t=5.212) (TFCE corrected, P<0.05, cluster size≥50 voxel). (3) Correlational analysis revealed that the negative correlations were found between PVT task performance and the FA value in the right superior longitudinal fasciculus( r=-0.492, P<0.001), right anterior limb of internal capsule( r=-0.510, P<0.001), right posterior thalamic radiation( r=-0.502, P<0.001) and body of corpus callosum( r=-0.464, P<0.001). The positive correlations were found between PVT task performance and the MD value in the body of corpus callosum( r=0.500, P<0.001), right superior longitudinal fasciculus( r=0.499, P<0.001), splenium of corpus callosum( r=0.462, P<0.001), right anterior limb of internal capsule( r=0.471, P<0.001), and AD value in right superior longitudinal fasciculus( r=0.643, P<0.001), as well as RD value in right superior corona radiate( r=0.498, P<0.001) (Bonferroni corrected, P<0.003). Conclusion:Differences in the microstructural characteristics of white matter fiber tracts in specific brain regions may constitute the potential neuropathological basis for the phenotypes of vulnerable and resistant individuals to continuous attention after sleep deprivation.

OBJECTIVES: To determine the association between glycosylated hemoglobin (HbA1c) and ambulatory blood pressure or heart rate in hypertensive patients. METHODS: A total of 585 patients, who performed ambulatory blood pressure monitoring (ABPM) from September 2018 to April 2019 in Xiangya Hospital, Central South University, were enrolled and assigned into 2 groups (470 in a hypertensive group and 115 in a normal group). HbA1c levels were compared. According to the HbA1c level, the hypertensive group was divided into 2 subgroups: A high HbA1c group (HbA1c≥6.1%, RESULTS: The hypertensive group had higher HbA1c level than the normal group [(6.1±1.3)% vs (5.1±1.7)%, CONCLUSIONS In hypertensive patients, HbA1c is positively correlated with ambulate blood pressure, blood pressure load, and heart rate, and it has no correlation with blood pressure variability, heart rate variability, or morning blood pressure.
Blood Pressure ; Blood Pressure Monitoring, Ambulatory ; Glycated Hemoglobin A/analysis* ; Heart Rate ; Humans ; Hypertension