Objective To explore the therapeutic effect of cerebrospinal fluid (CSF) replacement combined with intrathecal injection on tuberculous meningitis. Methods Fifty-five patients with tuberculous meningitis were randomly divided into treatment group and control group. The patients in the treatment group were treated with CSF replacement combined with intrathecal injection on the basis of routine anti-tuberculosis therapy. The curative effects between these two groups were analyzed after 8 weeks. Results The total effective rate was 95.7% in the treatment group,compared with that 61.9% in the control group (P < 0.01); the healing rate in the treatment group was 56.2% vs 28.6% in the control group. In addition,the decay rate for WBC and CSF pressure in the treatment group are more rapid than those in the control group(P < 0.05). Conclusion CSF replacement combined with intrathecal injection has better curative effect in patients with tuberculous meningitis.

Objective To explore sinus heart rate turbulence (HRT) in patients with different subtypes of essential hypertension (EH), and analyze the relationship between HRT and autonomic nervous system function damage in these patients. Methods The study consisted of 107 patients with EH (EH group) and 46 controls (control group). Based on 24 hours dynamic electrocardiogram, all patients were divided into dipper,non-dipper,and anti-dipper blood pressure group. The indexes about HRT and heart rate variability (HRV) among these groups were calculated and compared,and the relationship between turbulence onset (TO),turbulence slope (TS) and 24 hours mean systolic blood pressure,diastolic blood pressure was analyzed. Results There were significant differences in TO,TS,SDNN between EH group and control group(P ＜ 0.05 ). TO in non-dipper and anti-dipper blood pressure group was significantly higher than that in control group( P ＜ 0.05 ), and TS was lower than that in control group(P ＜ 0.05 ). There was no significant difference in TO,TS between dipper blood pressure group and control group (P ＞ 0.05). TO in non-dipper and anti-dipper blood pressure group was significantly higher than that in dipper blood pressure group, but TS was lower than that in dipper blood pressure group (P ＜0.05). Correlation analysis showed that TO had positive relationship with average heart rate and age (rs = 0.265, P = 0.004;rs = 0.217, P = 0.018 ), but had negative correlation with SDNN and left ventricular ejection fraction (LVEF) (rx = -0.287,P = 0.002;rx =-0. 179, P = 0.049). Whereas, TS had negative correlation with average heart rate and age (r = -0.335, P =0.015 ;r = -0.238,P= 0.009), but had positive relationship with SDNN and LVEF(r = 0.540,P = 0.001 ;r =0.432,P = 0.001 ). Conclusions HRT of EH patients becomes significantly low. It suggests that the autonomic nerve function in EH patients be injured seriously.

Objective To investigate the diversify and clinical significance of tumour necrosis factor-alpha (TNF-α), immune globulin (Ig), adenosine deaminase (ADA) in cercbrospinal fluid (CSF) of patients with intracranial infection. Method The levels of TNF-α,Ig and ADA in CSF of 25 cases of purulent meningitis ,30 eases of cerebral tuberculosis ,28 cases of virus meningitis and 24 cases of controls were detected by ELISA, immunity velocity scattering nepbelometry and enzyme coupling method respectively. Results The levels of IgA, IgM, IgG, ADA, TNF-α were (41.72±11.31) mg/L(18.11± 2.62)mg/L, (181.60±41.19)mg/L, (13.41±3.42)U/L, (418.62±43.16)ng/L in cerebral tuberculosis patients. The levels of IgA, IgM, IgG, ADA and TNF-αwere (20.65±8.85)mg/L, (93.20±4.30)mg/L, (92.77±35.09)mg/L, (3.32±2.41) U/L, (476.93±45.16) ng/L in purulent meningitis patients, and those were (7.11±2.23)rag/L,(5.81±1.19)mg/L,(20.71±10. 54)mg/L,(2. 36±0. 44)U/L,(375.06±45.21) ng/L in virus meningitis patients. The levels of IgM,IgG and IgA in cerebral tuberculosis patients and purulent meningitis patients were significantly higher than those in virus meningitis patients and controls (P< 0.01). The levels of IgG,IgA heightened most markedly in cerebral tuberculosis patients. The activity of ADA in cerebra] tuberculosis patients was higher markedly than that in controls and the other patients(P< 0.01). The levels of TNF-α in purulent meningitis patients were higher than those in controls and virus meningitis patients(P< 0.05). The levels of TNF-α in purulent meningitis patients were the highest, and the next was in cerebral tuberculosis patients. But there was no significant difference of each index between viral meningitis patients and controls. Conclusions Detecting the activity of ADA in CSF is the most valuable in diagnosing cerebral tuberculosis. Synchronized detection of TNF-α,Ig, ADA may be have better clinical application in diagnosing intracranial infection.

Objective To investigate the efficacy of recombinent human B-type natriuretic peptide (rhBNP) in peripartum cardiomyopathy (PPCM) with refractory heart failure (RHF).Methods From January 2010 to January 2014,a total of 61 patients with PPCM-RHF of the First Hospital of Qinhuangdao were enrolled into this study.All patients were randomly assigned into the rhBNP group (30 cases) and control group (31 cases).rhBNP was given 1.5 μg/kg,2 mim intravenously in rhBNP group and then infused intravenously 0.007 5-0.030 μg/(kg · min) for 24 hours,0.015-0.030 μg/(kg · min) for 24 hours when systolic blood pressure(SBP) ≥ 85 mmHg (1 mmHg =0.133 kPa) and mean brachial arterial pressure (MBP) ≥ 65 mmHg.Routine therapy followed the clinic practice was administrated in control group for 24 hours.Recorded the dyspnea change.Blood samples were collected at different time points to investigate BNP at 6 h,14 d and 30 d.Ultrasonic cardiography (UCG) was performed to evaluate left ventricular ejection fraction (LVEF) and left ventricular end-diastolic dimension(LVEDD) before the initiation of the infusion,at day 1,14 and 30 after the infusion.All patients were followed up for one month to record the main adverse cardiac events (MACE),including heart failure recurrence,cardiac death and ventricular fibrillation and tachycardia.Results The time of dyspnea resolved in rhBNP group was significantly shorter than control group ((1.69 ± 1.07) h vs (2.69 ±1.39) h,P =0.002).Concentration of plasma BNP(6 h:(296.50±123.25) ng/L,14 d:(141.37±69.54) ng/L,30 d:(107.41±33.69) ng/L) was signifisantly reduced in both groups than basic line (727.07 =± 146.84) ng/L,and it was significantly different between two groups.LVEF raised and LVEDD decreased were observed at 24 hours((52.23±4.54) mm),14 d((49.60±4.20) mm) and 30 d((42.59±3.90) mm) in rhBNP group and were significant better than the control group ((56.33 ± 4.38) mm,(53.03 ± 4.95) mm,(48.85±4.96) mm;P ＜0.05).There was no significant difference in term of LVEF between two groups after treatment.However,24 h(4.12±41.13)％,14 d(4.10±43)％,30 d(44.52±3.43)％ were significantly higher than the baseline(36.73±5.82)％ in rhBNP group.MACEs were lower in rhBNP group at 30 dayscontrast to the control(10％ (3/30) vs.42％ (13/31);P =0.005).Conclusion Compared with conventional treatment,rhBNP can effectively improve the heart function in patients with PPCM-RHF,reduce the occurrence of major adverse cardiac events in 30 d,and improve the prognosis of patients.

Objective:To report the clinical features of KCNQ2-associated epilepsy and the novel mutations and unreported clinical phenotype of KCNQ2 gene, so as to provide help for treatment selection and prognosis evaluation.Methods:Among 979 patients with epilepsy and developmental delay who were admitted to the Department of Neurology,Children′s Hospital Affiliated to Capital Institute of Pediatrics from July 2015 to October 2019, a total of 13 patients were selected from 12 families with KCNQ2 gene mutation by whole exome sequencing technology. Suspected mutations were verified by Sanger sequencing on the probands and their parents to identify the source. The clinical phenotype and genotype were analyzed according to these results.Results:Among the 13 patients with epilepsy, the onset age of four cases were older than six months [two cases in infancy (epilepsy encephalopathy), one case in early childhood (epilepsy encephalopathy) and one case in adolescence (benign epilepsy)]. Eight cases were treated with oxcarbazepine, of whom five cases were seizure free, and two cases showed partial response (>50%). Two cases treated with topiramate were seizure free. Five novel mutations were found in this research, including c.379T>G(p.Y127D), c.1A>C(initial codon mutation), c.708G>C(p.W236C), c.1027G>T(p.A343S) and c.1649T>G(p.V550G).Conclusions:Although it was rare in clinical work, the variation of KCNQ2 gene existed in patients with childhood-onset epilepsy and adolescent-onset epilepsy. Meanwhile, five novel mutations of KCNQ2 gene were reported, which further expanded its gene spectrum. This research supported that oxcarbazepine was the efficient medicine for the KCNQ2-associated epilepsy. Genetic testing showed great help to the treatment of epilepsy.

Objective To construct a lentiviral vector carrying angiopoietin-1 (Ang-1) and DsRed gene, and to package a virus particles. Methods The Ang-1 lentiviral vector with DsRed (PLV.Ex3d.P/puro-CMV>Ang-1>IRES/DsRed-Express2) was constructed by Gateway technology, and identiifed by PCR and gene sequencing. The lentiviral vector was mixed with helper vector pLV/helper-SL3, pLV/helper-SL4 and pLV/helper-SL5 by Lipofectamine 2000 to prepare DNA-Lipofectamine?2000 complexes. The complexes were then added to transfect 293FT cells and package virus. The virus titers and infection ef-ifciency were determined by lfuorescence expression. Results Ang-1 lentiviral vector PLV.Ex3d.P/puro-CMV>Ang-1>IRES/DsRed-Express2 was constructed successfully as identified by PCR and gene sequencing. Lentivirus with high-efficiency infection was produced by transfection to 293FT cells and the virus titer was 5×108 TU/ml. Conclusions The recombinant len-tiviral vector for Ang-1 was successfully constructed by Gateway technology and the lentivirus with high-efifciency infection packaging can be used for further experiment of Ang-1 gene.

Objective To evaluate the impact of different doses of recombinant human B-type natriuretic peptid (rh-BNP) within the dosage of clinical rage on oxygen consumption during acute myocardial infarction (AMI) with heart failure (HF). Methods AMI-HF model of York pig was established by occluding coronary artery with balloon combined with in-jecting microthrombus. Then animals were randomized into rhBNP group and control group. Clinical dose of rhBNP ( 1.5μg/kg bolus followed by a continuous infusion with speed of 0.01, 0.02 and 0.03μg · kg-1 · min-1 for 60 minutes respectively in turn) was administrated in rhBNP group while equal volume of saline was given in the control group. Myocardial oxygen up-take (MOU) was measured by drawing blood from coronary artery and coronary sinus using a catheter. Coronary diameter was determined using quantitative coronary angiography. The observation points were at baseline (T0), instant after the mod-el establishment (T1), 60 min after continuous rhBNP infusion of 0.01, 0.02, 0.03μg·kg-1·min-1 (T2-T4) respectively. Re-sults Compared with the control group, pulmonary capillary wedge pressure, central venous pressure, heart rate, systolic blood pressure and MOU were significantly decreased after rhBNP administration. And cardiac output and coronary diame-ter were obviously increased with addition of rhBNP. There is a interaction of drug intervention and time. In rhBNP group, MOU was significantly decreased with drug administraion (T2-T4 vs T1,mL O2/L: 10.61 ± 0.35,9.85 ± 0.60,9.79 ± 0.31 vs 11.59 ± 0.37). Conclusion Intravenous administration of rhBNP in AMI-HF model could decrease MOU and PCWP while increase the cardiac output.

A simple, rapid and sensitive colorimetric reverse-transcription loop-mediated isothermal amplification (RT-LAMP) was developed for rapid detection of Middle East respiratory syndrome coronavirus (MERS-CoV). The method employed six primers that recognized sequences of a nucleocapsid gene for amplification of nucleic acids under isothermal conditions at 63 degrees C for 60 min. Products were detected through a LA-320c Loopamp Turbidimeter (real-time RT-LAMP) or visual inspection of color change by pre-addition of Hydroxynaphthol Blue dye (visual RT-LAMP). Specificity of RT-LAMP was validated by detection of several human coronaviruses and common respiratory viruses. MERS-CoV real-time RT-LAMP had a linear correlation (R2) of 0.995 at 10(3)-10(6) copies. The limit of detection of real-time RT-LAMP, visual RT-LAMP and quantitative real-time PCR was 500, 1000 and 100 copies/reaction, respectively. The established RT-LAMP assay was demonstrated to be a rapid screening tool for MERS-CoV infection, and could be suitable in resource-limited clinical sites and for field studies.
Coronavirus Infections ; virology ; DNA Primers ; genetics ; Humans ; Middle East Respiratory Syndrome Coronavirus ; classification ; genetics ; isolation & purification ; Nucleic Acid Amplification Techniques ; methods ; Reverse Transcription

Strong promoters might not be optimal to obtain maximum metabolic flux towards desired products, whereas modulating gene expression with multiple regulatory parts is an option to obtain optimal expression strength. Therefore, we assessed the difference of impact on beta-carotene production between modulating isoprenoid gene expression with multiple regulatory parts and strong promoter, to improve beta-carotene production through combined modulation of essential isoprenoid genes. Eight isoprenoid genes were modulated with six artificial regulatory parts having a wide range of strengths to assess their effects on beta-carotene production. Optimal strength for each isoprenoid gene expression was identified, leading to 1.2 to 3.5-fold increase in beta-carotene production. In contrast to previous reports, our work suggests that modulating dxr, ispG and ispH genes with appropriate strengths increase beta-carotene production. Beta-carotene yield reached 17.59 mg/g after combined modulation of dxs and idi genes, 8-fold higher than that of the parent strain. Modulating gene expression with multiple regulatory parts was better than strong promoter, providing a new gene modulation strategy for targeted biosynthesis.
Escherichia coli ; genetics ; metabolism ; Gene Expression Regulation ; Promoter Regions, Genetic ; Terpenes ; metabolism ; beta Carotene ; biosynthesis

Objective To evaluate the method of establishment of a minipig model of ischemic heart failure(HF) with acute myocardial infarction(AMI) by coronary balloon occlusion and coadministration of injecting of microthrombi and plastic microspheres.Methods A total of eighteen minipigs were selected.After coronary angiography,angioplasty balloons were placed in the mid-distal of left anterior descending(LAD).The balloon was inflated intermittently to occlude the LAD 3 times and then to occlude it continuously for 120 minutes.After the balloon was taken out,4F Judkins-type angiogrphic catheter was superelectively engaged in LAD and 3 mL intermixture of mierothrombi and plastic microspheres were injected at 10 minites interval until TIMI myocardial perfusion was grade<2 and left ventfieular end-diastolic pressure was maintained from 15 to 18 mmHg.Electrocardiogram(ECG),hemodynamic perameters,ultrasonic cardiogram,cTnI and CK-MB were measured.Myocardial infarction area was evaluated by histopathology.Results Fourteen days later,fifteen minipigs survived and fourteen satisfied the criteria(pulmonary capillary wedge pressure.PCWP>18 mmHg and eardio output (CO) decreased beyond 30% ). The changes of ECG, hemodynamic perameters, CKMB, cTnI and cardiac pathologic examination were in accordance with AMI. Conclusion A stable experimental method of establishment of minipig model of ischemic heart failure (HF) with acute myocardial infarction (AMI) by coronary balloon occlusion and coadministration of injecting of microthrombi and plastic mierospheres is succeded. This method has advantages such as closed chest, higher success rate and stability compared with the drug induced, taehycardia-pacing induced, coronary artery ligation induced or microsphere injection alone methods.