The Split-Cre system consists of two inactive polypeptides: NCre and CCre, which can be recombined into an active full-length Cre under certain conditions. This system is typically used with LoxP. To develop an efficient Split-Cre system, this study used Rma intein from Rhodothermus marinus to split Cre and screened out the split site S102 which could efficiently mediate the recombination of Cre in the "Traffic Light" reporter cell line. Moreover, the S102 Split-Cre system was delivered to mice by dual-adeno-associated virus (AAV), and it was demonstrated that the efficiency of the Rma intein-mediated S102 Split-Cre system was comparable to the full-length Cre in mice. This system lays a foundation for both basic and applied research on Split-Cre.
Inteins/genetics* ; Animals ; Integrases/biosynthesis* ; Mice ; Dependovirus/metabolism* ; Bacterial Proteins/genetics* ; Recombination, Genetic ; Humans

Objective To investigate the effect of proanthocyanidins(PC)on the neurite outgrowth of rat dorsal root ganglion(DRG)neurons.Methods In vitro,primary rat DRG neurons were cultured wtih a series of concenteation of PC to assess the effect of PC on the number and length of neurites as well as the morphology of growth cone.In vivo,the expression of growth associated protein 43(GAP43)in the early stage of injury was detected using the sciatic nerve crush model.Finally,the impact of PC on nerve growth factor(NGF)expression in DRG neurons was evaluated in vitro using immunofluorescence and ELISA.Results PC significantly increased the number and length of neurites and the number of pseudopodium in growth cones of DRG neurons.PC also promoted the expres-sion of GAP43 in the early stage of sciatic nerve injury in rats and enhanced the expression of NGF in DRG neurons.Conclusion PC may promote the neurite outgrowth by increasing the expression of NGF in DRG neurons.

Ischemia reperfusion (I/R) injury is caused by ischemic shock, cardiac arrest, resection and transplantation, and its mechanism is closely related to calcium overload. Mitochondrial calcium uniporter (MCU) is a highly selective calcium channel located in the mitochondrial inner membrane (IMM), mediating calcium ions into the mitochondrial matrix. The MCU complex with the MCU as the core plays an important role in maintaining calcium ion homeostasis and alleviating I/R injury in the heart, kidney, brain, liver and other organs. The mitochondria associated endoplasmic reticulum membranes (MAMs) is a key protein in this process.

Objective:To analyze genioglossus (GG) activation responses to the negative pressure of upper airway cavity during awake and different sleep stages in patients with different obstructive sleep apnea (OSA) graduation.Methods:This prospective cohort study started from August 2019 to January 2021, recruited 42 male OSA patients aged from 21 to 59 (38.77±8.42) years. After completing whole night polysomnography (PSG) and upper airway CT, each subject underwent drug-induced sleep with simultaneous monitoring of genioglossal electromyography (GGEMG) and pressure of epiglottis (P epi). Subjects were divided into three groups of mild OSA(7 males), moderate OSA(12 males), and severe OSA(23 males). The differences in upper airway CT measurements, parameters of GGEMG and P epi during awake and induced sleep were compared. Statistical analysis was conducted by SPSS 21.0. Results:There was no significant difference in the GGEMG parameters between the mild and moderate groups. In wakefulness, the peak phasic GGEMG of the severe group was higher than the mild group ( t=1.249, P=0.025), with no statistically difference in the corresponding P epi. In the sleep onset, the GGEMG parameters and P epi in severe group were higher than the other two groups. Linear regression analysis of the maximum GGEMG and maximum P epi at the end of obstructive apnea (OA) in all moderate plus severe patients ( n=35) was shown nonlinear correlation ( r=0.28, P=0.694). The airway length of the glossopharyngeal cavity was linearly correlated with the maximum P epi of OA ( r=0.468, R2=0.219, P=0.005). Conclusions:The individual difference of GG activation in OSA patients is related to the severity of the disease (frequency of respiratory events) and negative pressure stimulation. In moderate and severe OSA patients, GG activity is not in harmony with the corresponding negative pressure stimulation, which may be one of the mechanisms leading to the aggravation of OSA.

Objective: Buyinqianzheng Formula (BYQZF) is clinically employed in traditional Chinese medicine to treat Parkinson's disease (PD) by improving mitochondrial dysfunction. However, the underlying mechanisms by which BYQZF affects mitochondrial morphology remain unknown. Therefore, we observed the effects of BYQZF on mitochondria from the perspective of the PINK1/Parkin pathway. Methods: Cell survival rates were assessed by Cell Counting Kit-8 assay. Expression levels of PINK1 and Parkin mRNA were examined by qRT-PCR. Protein expression levels of PINK1, PINK1-Ser228, Parkin, Parkin-Ser65, Drp1, and Drp1-Ser637 were examined by western blotting. PINK1, Parkin, and Mito-Tracker? Red CMXRos (MTR) were stained by triple-labeled immunofluorescence, and observed under laser confocal microscopy. Results: Cell survival rate, mitochondrial form factor, mean length and number of mitochondrial network branches, mitochondrial activity, mRNA expression levels of PINK1 and Parkin, and protein expression levels of PINK1, Parkin, and Drp1-Ser637 were reduced after 1-methyl-4-phenylpyridinium (MPP+) intervention. In contrast, Pearson's correlation coefficients between PINK1 and Parkin, and between Parkin and MTR, as well as protein expression levels of PINK1-Ser228, Parkin-Ser65, and Drp1 increased significantly after MPP+intervention. Treatment with BYQZF increased cell survival rate, mitochondrial form factor, mean length and number of mitochondrial network branches, mitochondrial activity, mRNA expression levels of PINK1 and Parkin, and expression of PINK1, Parkin, and Drp1-Ser637 proteins. Pearson's correlation coefficients between PINK1 and Parkin, and between Parkin and MTR, as well as protein expression levels of PINK1-Ser228, Parkin-Ser65, and Drp1 decreased after BYQZF treatment. Conclusion: These results demonstrate that BYQZF has a protective effect on mitochondrial molecular mechanisms in the PD cell model, and the mechanism is related to the PINK1/Parkin pathway.

The clinical manifestations of patients with schizophrenia and patients with depression not only have a certain similarity, but also change with the patient's mood, and thus lead to misdiagnosis in clinical diagnosis. Electroencephalogram (EEG) analysis provides an important reference and objective basis for accurate differentiation and diagnosis between patients with schizophrenia and patients with depression. In order to solve the problem of misdiagnosis between patients with schizophrenia and patients with depression, and to improve the accuracy of the classification and diagnosis of these two diseases, in this study we extracted the resting-state EEG features from 100 patients with depression and 100 patients with schizophrenia, including information entropy, sample entropy and approximate entropy, statistical properties feature and relative power spectral density (rPSD) of each EEG rhythm (δ, θ, α, β). Then feature vectors were formed to classify these two types of patients using the support vector machine (SVM) and the naive Bayes (NB) classifier. Experimental results indicate that: ① The rPSD feature vector performs the best in classification, achieving an average accuracy of 84.2% and a highest accuracy of 86.3%; ② The accuracy of SVM is obviously better than that of NB; ③ For the rPSD of each rhythm, the β rhythm performs the best with the highest accuracy of 76%; ④ Electrodes with large feature weight are mainly concentrated in the frontal lobe and parietal lobe. The results of this study indicate that the rPSD feature vector in conjunction with SVM can effectively distinguish depression and schizophrenia, and can also play an auxiliary role in the relevant clinical diagnosis.
Bayes Theorem ; Depression ; Electroencephalography ; Humans ; Schizophrenia ; Signal Processing, Computer-Assisted ; Support Vector Machine

Objective: To compare the parameters of polysomnography (PSG) in sleep structure and respiratory events between dexmedetomidine-induced sleep and natural sleep. Methods: From April 2016 to September 2018, a total of 44 patients with obstructive sleep apnea (OSA) and 3 patients with simple snoring completed PSG monitor both in natural sleep and dexmedetomidine-induced sleep in Department of Otorhinolaryngology Head and Neck Surgery, Beijing Tsinghua Changgung Hospital. The PSG parameters were statistically analysed with SPSS 22.0 software. Results: The average dose of dexmedetomidine was (104.60±27.93) μg, and there was no significant difference between the induced-sleep efficiency and the natural sleep efficiency (82.14%±16.66% vs. 86.50%±9.18%, t=-1.559, P>0.05). There was no rapid eye movement(REM) stages in all 47 subjects and only 1 case of them had non-rapid eye movement(NREM) stage 3 in induced sleep. The percentage of NREM1 in total sleep time was statistically different between the two groups (42.10%±26.71% vs. 17.47%±11.68%, t=5.997, P<0.001),but there was no significant difference in the percentage of NREM2 in total sleep time between the two groups (56.96%±26.0% vs. 62.95%±9.03%, t=-1.521, P=0.135). About respiratory events, there were significant differences in apnea hypopnea index ((46.29±20.23)/h vs. (39.67±25.41)/h), obstructive apnea index (25.20[10.50,45.40]/h vs. 16.20[3.30,35.20]/h) between induced-sleep and natural sleep (t=2.297, Z=-3.008, all P<0.05), these difference were more significant in mild-to-moderate OSA. There were no statistically significant differences in central apnea index (0.00[0.00,2.80]/h vs. 0.40[0.10,1.20]/h), mixed apnea index (0.00[0.00,6.20]/h vs. 0.00[0.00,3.40]/h, hypopnea index (4.20[0.00,3.30]/h vs. 12.00[5.20,17.40]/h), Z=-0.110,-0.508,-1.544, all P>0.05). There were statistical differences in the lowest oxygen saturation (84.77%±7. 59% vs. 80.21%±11.62%, t=2.558, P=0.014). Conclusions There is no significant difference in sleep efficiency and NREM2 between dexmedetomidine induced sleep and natural sleep.NREM3 sleep is rare induced, but REM sleep is none of all. And dexmedetomidine induced sleep may aggravate obstructive sleep apnea, but not central apnea.

Objective: To determine the objective effects of adenotonsillectomy on pediatric obstructive sleep apnea hypopnea syndrome (OSAHS) through analyzing the polysomnography (PSG) results between pre and post-operation. Methods: A total of 56 pediatric OSAHS patients were included who underwent adenoidectomy or/and tonsillectomy and completed PSG follow-up from January 1, 2017 to March 31, 2018. All the pediatric patients who underwent adenoidectomy or/and tonsillectomy during the research period were arranged to take a preoperative PSG study. Patients who were diagnosed OSAHS would be encouraged to complete a follow-up PSG study ranged from1 to 3 months after surgery. The parameters of respiration and sleep architecture of PSG were compared and analyzed. The paired student t test was used to compare preoperative and postoperative mean values. The unpaired student t test was used to compare quantitative variables among different groups. The rank sum test was used if the data were abnormal distribution. Results: Totally 238 patients completed preoperative PSG study, 62 patients were diagnosed as pediatric OSAHS, 56 eligible patients finished post-operative PSG. Hypopnea was the majority in all type of respiratory events in 56.45% (35/62) subjects, while central apnea as the majority in 29.03% (18/62) subjects who can also get significant CAI decrease after surgery. However, obstructive apnea as the majority only exist in 14.52% (9/62) subjects. The short-term cure rate of pediatric OSAHS was 85.71% (48/56). The postoperative AHI, MAI, CAI, HI, ODI, LoSpO₂, percentage of stage I sleep and arousal index were significantly decreased, however, the OAI was no statistical decrease. The percentage of stage Ⅱ and rapid eye movement (REM) sleep were significantly increased, while no significant change in percentage of slow wave sleep and sleep efficiency(t=2.32, P=0.017). Conclusions Pediatric OSAHS manifest different characteristics of respiratory events from that of adults. Adenotonsillectomy can significant decrease respiratory events and improve sleep architecture, however, there are still some patients who can′t be completely relieved with adenotonsillectomy.

In the paper,we introduced the peculiarity of Candida albicans and the disease caused by it,expounded the complexity of the pathogenesis,enumerated the advantages of the RNA-Seq and reviewed its application to study on the pathogenesis of Candida albicans,found out some shortages in previous studies,and anticipated the possible trends of such study in future.In conclusion,some remarkable achievements will bring about by use of improved RNA-Seq for intensive researches on the pathogenesis of Candida albicans.

BACKGROUND: Thoracolumbar segments (T11-L2) prone to damage due to its special anatomical and biomechanical characteristics. Therefore, fully understanding the shape of pedicle of vertebral arch and finding the visible, constant, and the point of insertion of the pedicle axis is very important to the safety of pedicle screw placement.OBJECTIVE: To measure the surgically relevant parameters of thoracolumbar pedicles between Han and Uygur males using computed tomography (CT) scan to provide some anatomic reference data for pedicle screw fixation.METHODS: The vertebral bodies and pedicles of adult males were scanned (60 cases of Han people and 60 cases of Uygur people) from T10-L3 with CT. The parameters were processed by three-dimensional reconstruction. Transverse pedicle width, pedicle axis length, transverse pedicle angle, and sagittal pedicle angle were measured by using length and angle measurement tool of browser in bone-window CT images. The age and stature information were recorded. All the data above were processed by SPSS 13.0 software.RESULTS AND CONCLUSION: (1) The mean transverse pedicle width of T12 and L1 in Han was bigger than that in the Uygur. (2) The mean transverse pedicle angle of T12 and L2 in Han was bigger than that in the Uygur. (3) The mean pedicle axis length and sagittal pedicle angle of T11 and L1 in Han were bigger than that in the Uygur (P < 0.05). (4) There were some differences among some parameters of the thoracolumbar pedicles between the Han and Uygur people.Data from any study only can be used as a guide for pedicle screw fixation. Preoperative CT evaluation may provide an individualized strategy to reduce the incidence of postoperative complications caused by misplacement.