To construct CRISPR/Cas9 vectors for the editing of SmPAL1 in the phenylpropane metabolic pathway of Salvia miltiorrhiza, CIRSPR/Cas9 target sites of SmPAL1 were designed by online software. Its target efficiencies were detected in vitro by enzyme digestion and sequences with highly efficiency were constructed into CRISPR/Cas9 vectors. Three possible CRISPR target sequences (SmPAL1-g1, SmPAL1-g2, SmPAL1-g3) were designed and the enzyme digestion efficiencies were 53.3%, 76.6% and 10.0%. SmPAL1-g1 and SmPAL1-g2 were constructed into vector VK005-03 named as VK005-03-g1 and VK005-03-g2. The results of sequencing showed that the two CRISPR/Cas target sequences were all constructed into VK005-03. Here we first laid the foundation for the study of SmPAL1 and provided an effective strategy for the screening of sgRNA.
CRISPR-Cas Systems ; Clustered Regularly Interspaced Short Palindromic Repeats ; Metabolic Networks and Pathways ; genetics ; Salvia miltiorrhiza ; chemistry ; genetics

It is necessary to improve the stability of human serum albumin in response to the complex temperature, light and other conditions during the manufacture and storage. In this paper, the stabilization effect and simple stabilization mechanism of ligands on albumin were described from the perspective of ligand binding to albumin.Through review and comparison, it can be concluded that the common ligand sodium octanoate mainly plays a role in improving thermal stability, and the common ligand N-acetyl-L-tryptophan mainly plays a role in improving antioxidant activity, N-acetyl-L-methionine has better antioxidant and anti-photooxidation than N-acetyl-L-tryptophan.

Based on analyzing image edge cloning theory and algorithms, a marginal clone algorithm is proposed with a combination of edge connectivity and noise removal algorithm. Simulation results show that the edge image detected by the algorithm has solved problems of traditional edge discontinuities and tsereve noise. Moreover, it has better edge recognition performance.
Algorithms ; Humans ; Image Enhancement ; methods ; Retinal Vessels

The Portable dynamic state electrocardiogram collecting system is introduced by using TMS302VC5402, TLC320AD50C, liquid crystal display model, and so on. This dissertation describes the work principle of the system and uses the united algorithm based on wavelet to identify and locate the ECG characteristic waves. This system has as follows of advantages: big memory, low noise,high common mode rejection ratio, the low power consume,the long record time etc.
Algorithms ; Electrocardiography, Ambulatory ; instrumentation ; Equipment Design ; Signal Processing, Computer-Assisted ; Wavelet Analysis

The purpose of the present study was to investigate the effect of advanced glycated albumin (AGE-alb) on the activation of caspase-12, a key molecule in endoplasmic reticulum stress (ERS)-associated apoptotic pathway, and to elucidate the underlying molecular mechanisms of macrophage apoptosis. RAW264.7 macrophages were treated with AGE-alb (2, 4 and 6 g/L), control albumin (C-alb, 4 g/L), tunicamycin (TM, 4 mg/L), or pretreated with 4-phenylbutyric acid (PBA, 5 mmol/L) for 1 h and then treated with AGE-alb (4 g/L). After incubation for 24 h, the cell viability and apoptosis were determined by using MTT assay and TUNEL detection kit, respectively. Lactate dehydrogenase (LDH) activity in media was determined by using an assay kit. The protein levels of caspase-12 were examined by Western blot analysis. The results showed that like TM (an ERS inducer), incubation with AGE-alb led to significant decrease in viability and increase in LDH activity in media and apoptotic rate in a dose-dependent manner. In addition, AGE-alb induced activation of caspase-12 especially at the concentration of 4 and 6 g/L (P < 0.01), which was similar to TM. However, PBA (an ERS inhibitor) protected RAW264.7 macrophages from AGE-alb-induced decrease in viability and increases in LDH activity and apoptosis. Moreover, PBA also inhibited the caspase-12 activation induced by AGE-alb (P < 0.05). These results suggest that AGE-alb may induce apoptosis in RAW 264.7 macrophages, and the mechanism may be related to the activation of ERS-associated apoptotic pathway mediated by caspase-12.
Animals ; Apoptosis ; Caspase 12 ; Cell Line, Tumor ; Cell Survival ; Endoplasmic Reticulum Stress ; Macrophages ; Mice ; Phenylbutyrates ; Serum Albumin ; Tunicamycin

To establish a rapid method for analysis of gingerol-related compounds in fresh ginger by using high performance liquid chromatography coupled with electron spray ionization-quadrupole-time of flight mass/mass spectrometry (HPLC-ESI-Q-TOF-MS/ MS). The gingerol-related compounds in fresh ginger was separated by an Inertsil ODS-SP column (4.6 mm x 250 mm, 5 microm) using a binary eluent under gradient conditions. The analytes were detected by ESI-Q-TOF-MS/MS in positive ion mode to obtain MS and MS/MS spectra and to extract molecular weights. From the MS data, the accurate molecular weights of gingerol-related compounds were obtained, and from the MS/MS data, the (+) ESI-Q-TOF-MS/MS fragments were obtained. 25 gingerol-related compounds were identified from fresh ginger by attentive studying on the mass spectra of compounds and comparing with reference data reported in the literature, respectively. This method was certified to be accurate and reliable and can be used for the rapid analysis of gingerol-related compounds in fresh ginger.
Catechols ; analysis ; Chromatography, High Pressure Liquid ; methods ; Fatty Alcohols ; analysis ; Ginger ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tandem Mass Spectrometry ; methods

Based on the reconstruction of two-dimension phase space of time series of short ECG signals, the variation of the strange attractor geometry is described and two indices, VMI and VAI, are derived in this paper. The two indices can distinguish clearly the ECG signals of sinus rhythm, tachycardia and ventricular fibrillation. Stable results of VMI and VAI can be obtained by analyzing ECG signals of several seconds. They are expected to be used in the development of medical instruments for a fast realtime display of analysis results.
Animals ; Electrocardiography ; methods ; Myocardial Infarction ; physiopathology ; Rabbits ; Signal Processing, Computer-Assisted

The genomic diversity of Avian leukosis virus subgroup J (ALV-J) was investigated in an experimentally infected chicken. ALV-J variants in tissues from four different organs of the same bird were re-isolated in DF-1 cells, and their gp85 gene was amplified and cloned. Ten clones from each organ were sequenced and compared with the original inoculum strain, NX0101. The minimum homology of each organ ranged from 96.7 to 97.6%, and the lowest homology between organs was only 94.9%, which was much lower than the 99.1% homology of inoculum NX0101, indicating high diversity of ALV-J, even within the same bird. The gp85 mutations from the left kidney, which contained tumors, and the right kidney, which was tumor-free, had higher non-synonymous to synonymous mutation ratios than those in the tumor-bearing liver and lungs. Additionally, the mutational sites of gp85 gene in the kidney were similar, and they differed from those in the liver and lung, implying that organ- or tissue-specific selective pressure had a greater influence on the evolution of ALV-J diversity. These results suggest that more ALV-J clones from different organs and tissues should be sequenced and compared to better understand viral evolution and molecular epidemiology in the field.
Animals ; Avian Leukosis Virus* ; Avian Leukosis* ; Birds ; Chickens* ; Clone Cells ; Kidney ; Liver ; Lung ; Molecular Epidemiology ; Silent Mutation

Objective:To explore the efficacy of cerebral vasospasm model established by brief double blood injection in cisternal cisteria.Methods:Twenty-five SD rats were randomly divided into sham-operated group, group of 1 d after subarachnoid hemorrhage (SAH), group of 3 d after SAH, group of 5 d after SAH, and group of 7 d after SAH ( n=5). Autologous blood (0.2 mL, obtained by caudal artery puncture) was directly injected into the atlanto-occipital membrane and repeated 48 h after that to establish cerebral vasospasm model. Neurological impairment was evaluated by modified Neurological Severity Score (mNSS). Diameter and cross-sectional area of the basilar artery (BA) were detected by HE staining. Differences of body mass before modeling, body mass between the 2 blood injections, mNSS scores, and diameter and cross-sectional area of BA were compared among groups. Results:(1) Body mass before modeling was not significantly different among the 5 groups ( P>0.05); differences of body mass between the 2 blood injections in the group of 1 d after SAH, group of 3 d after SAH, group of 5 d after SAH, and group of 7 d after SAH were significantly greater than those in the sham-operated group ( P<0.05). (2) The mNSS scores in the group of 1 d after SAH, group of 3 d after SAH, group of 5 d after SAH, and group of 7 d after SAH were significantly higher than those in the sham-operated group ( P<0.05). (3) BA diameter in the group of 3 d after SAH and group of 7 d after SAH was significantly shorter than that in the sham-operated group, and that in the group of 7 d after SAH was significantly shorter than that in the group of 1 d after SAH and group of 5 d after SAH ( P<0.05). BA cross-sectional area in the group of 1 d after SAH, group of 3 d after SAH, group of 5 d after SAH and group of 7 d after SAH was significantly smaller than that in the sham-operated group, and that in the group of 7 d after SAH was significantly smaller than that in the group of 1 d after SAH, group of 3 d after SAH and group of 5 d after SAH ( P<0.05). Conclusion:Compared with other traditional models, cerebral vasospasm model established by brief double blood injection in cisternal cisteria has advantages of simplified operation, short modeling time, and minimal invasion; model of 7 d after autologous blood injection enjoys optimal.

Objective:To investigate the CD163 expression characteristics in intracranial aneurysm (IA) tissue and blood of patients with IA and its feasibility as an early clinical screening indicator for IA.Methods:A total of 28 patients with IA admitted to Department of Neurosurgery, First Affiliated Hospital of Xinjiang Medical University from January 2021 to November 2023 were selected as IA group, and 28 healthy subjects from Health Management Center, First Affiliated Hospital of Xinjiang Medical University at the same time period were selected as control group. Eight saccular IA tissues and 12 superficial temporal artery tissues were collected from patients from IA group accepted IA clipping, and real-time fluorescent quantitative PCR (RT-qPCR) was used to detect the CD163 mRNA expression in these tissues. RT-qPCR was also used to detect the CD163 mRNA expression in the blood of the 2 groups. Seven patients with IA and 7 control subjects from the above 2 groups were randomly selected, respectively; and plasma CD163 protein content was detected by enzyme-linked immunosorbent assay (ELISA). Multivariate Logistic regression was used to analyze the influencing factors for IA. Receiver operating characteristic (ROC) curve was used to analyze the diagnostic values of blood CD163 mRNA expression and plasma CD163 protein content in IA. Results:CD163 mRNA expression in IA tissues was significantly higher than that in superficial temporal artery tissues (41.870±20.355 vs. 6.080±5.444, P<0.05). CD163 mRNA expression in the blood of IA patients was significantly higher than that in the controls (1.969[1.124, 2.318] vs. 1.124[0.933, 1.379], P<0.05). CD163 mRNA expression in the blood of ruptured IA group, unruptured IA group, and control group was gradually decreased, with significant differences ( P<0.05). CD163 mRNA expression in the blood of female IA patients was not statistically different compared with that in male IA patients ( P>0.05). ELISA showed that the CD163 protein content in plasma of the IA group was significantly higher than that in the control group [10.537±1.879] ng/L vs. [8.598±0.885] ng/L, P<0.05). Multivariate Logistic regression analysis showed that age and CD163 mRNA expression in the blood were independent influencing factors for IA occurrence ( OR=0.844, 95% CI: 0.750-0.951, P=0.005; OR=0.111, 95% CI: 0.024-0.506, P=0.004). ROC curve showed that the area under the curve (AUC) of CD163 mRNA expression in blood in diagnosing IA was 0.759 (95% CI: 0.618-0.890, P=0.002), and that of CD163 protein content in plasma in diagnosing IA was 0.864 (95% CI: 0.610-1.000, P=0.035). Conclusion:CD163 mRNA expressions in blood and IA tissues and CD163 protein content in plasma are high in patients with IA; CD163 mRNA expression in blood is an independent risk factor for IA; CD163 protein in plasma can be used as a molecular marker for screening IA.