The successful establishment of animal models of cardiac arrest-cardiopulmonary resuscitation (CA-CPR) undoubtedly provided an important basis for exploring the method of cardiopulmonary resuscitation (CPR) and advanced cardiovascular life support (ACLS). However, pathophysiology varied with the etiology of cardiac arrest (CA). Therefore, preparation of similar animal models according to etiology was the basis for pathophysiological changes research. Compared with other animals, the rabbits had both the advantages of large and small animals, so they became common research object for the CA-CPR model. This paper reviewed the common methods of animal models of CA-CPR in rabbits. In this review, the methods, criteria, advantages, disadvantages and precautions of each model were analyzed, which would provide useful reference for CPR researchers.

In the present study, packaging system composed of pAAV-CMV-GFP, pAAV-RC and pHelper were transfected into human embryonic kidney 293 cells (HEK293 cells) mediated by polyethyleneimine (PEI) to explore an optimal transfection condition. Different total plasmid DNA dosages (1, 2, 3, 4, 5, 6 μg) and different PEI/Plasmid ratios (1:1, 3:1, 5:1, 7:1) were tested with detection of green fluorescence protein (GFP) with ImagePro Plus6. 0 Software. Then transfection efficiency of the optimized transfection system was further observed for different time periods(12, 24, 36, 48, 60, 72 h). The results showed that total plasmid dosage of 4 μg/well with PEI/plasmid ratio of 3 : 1-5 : 1 was an efficient transfection condition. Transfection efficiency-time curve was an S-shaped curve. Transfection efficiency reached a plateau at 60 h after transfection. The optimized conditions for PEI-mediated transfection at the optimal time result in enhanced transfection efficiency of triple plasmid into HEK293 cells.
Green Fluorescent Proteins ; HEK293 Cells ; Humans ; Plasmids ; Polyethyleneimine ; Transfection ; methods

Iliac arteriovenous ifstula (AVF) usually manifests in a wide range of symptoms similar to typical deep venous thrombosis (DVT), which often lead to delayed diagnosis or misdiagnosis. We reported a 51-year old woman who was performed lumbar discectomy and showed a progressive abdominal distention, dyspnea, and swollen left leg. She was initially diagnosed as deep vein thrombosis and the ifnal diagnosis was arteriovenous ifstula. hTe ifstula was successfully sealed by an endovascular covered stent. No further recurrence was found atfer a half year’s follow-up. hTis article summarized the experience regarding iliac arteriovenous ifstula misdiagnosed, and discussed the differential diagnosis between arteriovenous ifstula and pulmonary thromboembolism causedby deep vein thrombosis.

Objective To estimate the predictive value of neutrophil gelatinase-associated lipocalin in urine (uNGAL) for detection of acute kidney injury (AKI) in the intensive care unit (ICU) critically ill patients.Methods A total of 110 patients from the ICU of three general hospitals were enrolled in the study.The patients were adults more than 18 years of age.After admitted to ICU,the patients were continuously observed for 72 hours.According to the RIFLE criteria for diagnosis of AKI,the patients were classified as AKI group (33 cases) or non-AKI (77 cases).According to the sepsis diagnostic criteria,the patients were classified as sepsis (79 cases) or non-sepsis (31 cases).Exclusion criteria of patients were chronic renal insufficiency,malignant tumor,death after admitted to ICU 24 hours.Serum creatinine and uNGAL of the patients were analyzed daily.The difference of uNGAL between sepsis and non-sepsis patients,AKI and non-AKI patients,sepsis non-AKI and sepsis AKI patients was compared.Moreover,the difference of serum creatinine and uNGAL between AKI and non-AKI patients into ICU 24 h was compared,and the sensitivity and specificity of uNGAL and serum creatinine for diagnosis of AKI in the ICU patients were evaluated using ROC curve.Results The uNGAL levels were all significantly different between sepsis and non-septis patients,AKI and non-AKI patients,sepsis concomitant AKI and sepsis without AKI patients.The uNGAL levels were significantly different between AKI and non-AKI patients in ICU for the first 24 h,while the difference of serum creatinine were not significant.The area under receiver operating characteristic (ROC) curve of uNGAL and serum creatinine of patients in ICU for the first 24 h were 0.828 (95％ CI:O.742-0.914) and 0.583 (95％ CI:0.471-0.695),respectively.The cutoff value of uNGAL was 170 ng/ml,and the sensitivity and specificity were 0.778 and 0.784,respectively.The sensitivity was superior to serum creatinine.Conclusions uNGAL was superior to serum creatinine in the diagnosis of AKI,and could be used as a marker of the early diagnosis of AKI.

Objective To study the changes of PI3K/Akt/GSK3β signaling pathway during resuscitation with neck cooling in order to explore the relationship between the protective effect of neck cooling and the phosphorylation of PI3K/Akt and GSK3β.Methods Thirty rabbits were randomly(random number) divided into five groups, and models of cadiac arrest were induced by ventricular fibrillation(VF, the positive electrode in the right ventricle and negative pole on the apex of heart) for 4 min.In sham group,a electrode was placed into right ventricle without electric current conducted, and CA was not induced.The rabbits were sacrificed and specimens were taken at 24 hours after modeling.In normothermia treat group(NT group),resuscitation was carried out to restoration of spontaneous circulation(ROSC),and the rabbits were sacrificed and specimens were taken at 24 hours after modeling.In intra-arrest therapeutic hypothermia group (IATH group), rapid neck cooling was initiated at the same time with CPR,and the target brain temperature was set at 34 ℃ maintained for 4 hours after ROSC.Rabbits were sacrificed and specimens were taken at 24 hours after modeling.In recovery period cooling + LY294002 group(PATH+LY294002 group), LY294002 was injected intra-ventricularly at 20 minutes before resuscitation.Rapid neck cooling was started at the same time with CPR,and the target brain temperature was set at 34 ℃ maintained for 4 hours after ROSC.The rabbits were sacrificed and specimens were taken at 24 hours after modeling.In post-arrest therapeutic hypothermia group (PATH group), rapid neck cooling was begun after CPR for 1 hour,and the target brain temperature was set at 34 ℃ maintained for 4 hours after ROSC.The rabbits were sacrificed and specimens were taken at 24 hours after modeling.Animals were sacrificed by using overdose anesthetic drug.Western blot was used to detect the level of Akt p-Akt GSK-3β p-GSK-3β (ser9) protein, and TUNEL was used to observe apoptosis of tissues in each group.Multiple comparisons were performed with one-way analysis of variance (ANOVA).Results Compared with Sham group, Akt (Thr-308) phosphorylation (P-AKT) and P-GSK-3β levels in the brain neuron cytoplasm in 24 hours after CPR resuscitation in NT group was significantly reduced, and showed a gradual reduction trend (P<0.05);the P-AKT and P-GSK-3β levels in the brain neuron cytoplasm in 24 hours after CPR resuscitation in IATH group were significantly enhanced compared with NT group (P<0.05);the levels of these two kinds of protein at one hour after resuscitation in PATH group were significantly enhanced compared with NT group (P<0.05), but lower in IATH group.Intra-ventricularly injection of LY294002 made the effect of hypothermia lost, indicating that LY294002 inhibited the phosphorylation of Akt.Apoptosis cells were significantly reduced in IATH group and normothermia theatment group compared with PATH group and LY294002 group(P<0.05).Conclusions Neck cooling can reduce apoptosis in rabbit brain cells after recovery, and the protective effect on brain is best in intra-arrest therapeutic hypothermia group.LY294002 specifically block the PI3K/Akt pathway, and the protective effect of cooling on the brain can be abolished,indicating hypothermia protects the neurological function via activation of PI3K/Akt pathway.Neck cooling protects the neurological function by activating PI3K/Akt/GSK-3β, promoting the Akt activation, and increasing the expression of P-GSK3β.Specific Akt inhibitor LY294002 inhibits Akt phosphorylation of brain tissue recovery and further inhibit the phosphorylation of GSK-3β, thus abolishing protective effect of cooling on neurological function.

Objective To investigate the role and neuroinflammatory mechanism of iron on dopamine ( DA) neurons in multiple primary midbrain cultures that mimic human substantia nigra pars compacta.Methods Ferrous chloride ( Fe2+ ) with the desired concentrations of 5,25 and 100 μmol/L was used to ( 1 ) treat primary midbrain neuron-microglia-astroglia cultures for 7 days and the numbers of DA neurons and total neurons were counted after tyrosine hydroxylase (TH) and neuron-specific neuclear protein neurons in 5,25 and 100 μmol/L Fe2 + -treated groups were 89%,70% and 55% of control group,and 25,100 μmol/L Fe2+ significantly decreased DA neuronal numbers compared with control group ( F = 12.047,P ＜0.01);DA neuronal bodies were shrunk and smaller,cytoplasmic stainings were reduced,neuronal dendrites were decreased;(2) The numbers of Neu-N ( + ) total neurons in 5,25 and 100 μmol/L Fe2+-treated groups were 100%,104% and 101% of control group and Fe2+ did not decrease DA neuronal numbers compared with control group (t =4.458,P ＞ 0.05 );5,25 and 100 μmol/L Fe2+-induced difference between total neurons and DA neurons were 11%,34% and 46%,and 25 and 100 (Amol/L Fe2+ produced significant difference(t =8.098,P ＜0.05;t = 11.218,P＜0.05);(3) In primary midbrain neuron-microglia-astroglia and neuron-astroglia cultures,the numbers of DA neurons in 25 μmol/L Fe+-treated group were 70% and 98% of control group,respectively.The difference between two groups was 28%,which was statistically significant (t =8.061,P＜0.05);The numbers of DA neurons in 100 μmol/L groups were 183%,190 % and 240% of control group,and 25 and 100 μmol/L Fe2 + significantly increased microglial numbers compared with control group ( F = 6.101,P ＜ 0.01 );dramatic changes of microglial morphology were indicated by the enlarged cell bodies and irregular shape.Conclusions Fe2 + provokes selective DA neuronal damage and microglia are the mediators of the neurotoxic effect,which may be due to microglial over-activation featured by the significant production of neurotoxic factors and morphological changes of microglia.This investigation cast a new light for PD therapy by inhibiting Fe2+ -induced neuroinflammation characterized by the microglial over-activation.

Objective:To investigate the vitamin D level of pulmonary tuberculosis patients in Qinghai Province, and to explore the correlation between vitamin D level and pulmonary tuberculosis.Methods:From May to September 2020, 208 bacterial confirmed pulmonary tuberculosis patients who were admitted to The 4th People′s Hospital of Qinghai Province were enrolled as the pulmonary tuberculosis group, and 129 healthy people who underwent physical examination during the same period were enrolled as the healthy control group. Independent sample t test and chi-square test were used for statistical analysis. Results:The deficiency rate of vitamin D was 11.06%(23/208) in the pulmonary tuberculosis group, which was higher than that (3.10%(4/129)) in the healthy control group, and the difference was statistically significant ( χ2=6.840, P=0.009). The vitamin D level was (56.84±20.03) μg/L in the pulmonary tuberculosis group, which was lower than that ((67.39±17.07) μg/L) in the healthy control group, and the difference was statistically significant ( t=5.154, P<0.01). The vitamin D levels were not different between the newly treated ((56.66±20.02) μg/L)) and retreated pulmonary tuberculosis patients ((59.11±20.81) μg/L) ( t=0.468, P=0.650). The vitamin D level of simple pulmonary tuberculosis patients ((57.82±20.01) μg/L) was higher than that of pulmonary tuberculosis patients combined with other diseases ((48.08±18.46) μg/L), and the difference was statistically significant ( t=2.132, P=0.034). Conclusion:Pulmonary tuberculosis is associated with decreased vitamin D levels, and patients with pulmonary tuberculosis are more likely to suffer from decreased or deficient vitamin D, which suggests clinicians considering the vitamin D status when treating pulmonary tuberculosis patients.

Objective To illustrate the semiological characteristics of the three sub-types within the broad bilateral asymmetric tonic seizures (BATS),summarize their predictive values on lateralization and localization of seizure onset zone (SOZ),and analyze the difference between BATS and asymmetrical tonic limb posturing (ATLP).Methods A retrospective review of 385 patients who underwent stereotactic electrode implantation in the Sanbo Brain Hospital,Capital Medical University from September 2011 to May 2018 was performed.As long as there was a clinical epileptic seizure in the presence of BATS or ATLP,the patients were classified into the corresponding groups.Postoperative prognosis was assessed using Engel's grading criteria for a follow-up of no less than six months.Seizure descriptions were based on the classification of epileptic seizures introduced by Lüiders,which used arrows to connect the symptoms in chronological order.Results There was no statistically significant difference between the classic BATS and bilateral proximal tonic seizure in terms of whether it could be an independent seizure,as the onset and end of the seizure,with version and generalized tonic-clonic seizure (P＞0.05).Compared with the ATLP,except for whether it could be an independent seizure (P=1.000) and onset before versive seizure (P=0.068),the BATS showed significantly different semiological features (P＜0.05).The classic BATS and secondary motor area epilepsy had a 100.0％ predictive accuracy on the lateralization of SOZ.In the patients with broad BATS,the SOZ distribution was more extensive,but it was rare in the orbitofrontal gyrus,frontal pole and mesial temporal lobe.Compared with the bilateral proximal tonic seizures from the other regions,those originated from supplementary somatosensory motor area and its adjacent areas were rare and showed no statistically significant difference (0/8 vs 40.0％ (18/45),x2=3.226,P=0.072) but a low trend.The predictive value of BATS on lateralization of SOZ was higher than that of ATLP (84.9％ (45/53) vs 57.1％ (24/42),x2=9.086,P=0.003),and BATS was less originated from temporal lobe than ATLP (3.8％ (2/53) vs 23.8％ (10/42),x2=8.523,P=0.004).Conclusion Different from ATLP,the broad BATS are characterized by tonic proximal upper limb posturing,and have a higher predictive value on lateralization and localization of SOZ.

The epileptic eye and head movements during epileptic seizures may be much more complicated thanpeople originally understood, which can be ipsilateral or contralateral to the electroencephalographyfocus. Here, we describe a male patient with drug resistant focal seizures associated with a directionalseparation between head and eye movement before evolving into generalized tonic-clonic seizure.His contralateral head leading turning showed forced, sustained, and unnatural features companiedby ipsilateral eye staring. Stereoeletroencephalography monitoring was performed, and 4 habitualseizures were recorded over 5 days. Three seizures showed left head leading turning and generalizedtonic-clonic seizure, and only one showed dizziness and ringing in the ears. All the seizures showedthat the ictal onset contacts were located in the posterior inferior temporal sulcus which borders onthe anterolateral part of medial superior temporal area. The patient underwent a resection includingtemporooccipital region, and the histopathology showed focal cortical dysplasia type Ic. He has beenseizure free for two years after operation. The scores of the intelligence and memory quotient improvedhalf year after operation.