Objective: To evaluate the clinical profile of target controlled infusion based anesthesia using remifentanil and propofol. Methods: 16 ASA I II patients undergoing elective laparoscopic cholecystectomy were enrolled. Remifentanil was set at 7?g?L -1 as target and propofol at 3 mg?L -1 throughout the whole procedure. The hemodynamics during induction of anesthesia and recovery profiles were recorded. Arterial blood samples for analysis of remifentanil were taken 15min after infusion, 20 min after infusion and at time of emergence. Results: After induction of anesthesia, systolic blood pressure (SBP) decreased from (144?27) mm Hg to (101?18) mm Hg ( P 0.05). SBP, MBP and HR remained stable after intubation for 3min. No patient showed haemodynamic stress to tracheal intubation. Times from stopping administration of anesthetics until full spontaneous respiration, eye opening, tracheal extubation, orientation and discharging from the postanesthetic care unit (PACU) were (12?6), (9?4), (13?6), (15?5) and (19?7) min respectively. Measured drug values of remifentanil were (4.6?9.5) ?g?L -1 , (6.6?11.5) ?g?L -1 , (1.2?8.7) ?g?L -1 respectively. Conclusion: Remifentanil/propofol TCI based anesthesia achieved the optimal hemodynamic stability during anesthesia induction and maintenance, and better recovery profile from anesthesia. Measured drug values of remifentanil showed a considerable interindividual variation and more lower than the set target.

Objective: To analyze Internet behaviors of children and adolescents and their influencing factors, and to provide scientific basis for health education guidance and prevention measures. Methods: A stratified cluster sampling method was used. From January to June 2019, 30 955 primary school students in 16 provinces (municipalities), from eastern, central, and western economic zones of China were selected to conduct a questionnaire survey, regarding general demographics, diet and exercise, tobacco, alcohol and sleep, Internet and electronics usage, etc. Results: Among them, 43.1% of students had never been online with girls (44.7%) higher than boys (41.8%)(χ2=21.04, P<0.01); 1.3% of students reported online time for more than 4 hours a day, boys were significantly higher than girls(χ2=15.87, P<0.01); Internet addiction was detected in 2.4% of elementary school students, with boys (3.0%) higher than girls(1.7%)(χ2=331.77, P<0.01), and the trend increases with grade. With sixth grade (3.3%) were significantly higher than children from the first grade (0.8%) and other grades(χ2=163.96, P<0.01). Children’s tendency to Internet addiction was 11.9%, of which boys were 13.0% and girls were 10.6%, boys were significantly higher than girls(χ2=117.66, P<0.01). Pearson correlation analysis revealed that, after controlling gender age, boarding school, family relationship, single parents and regrouped families, smoking, drinking sugary drinks, eating fried foods, and left-behind were positively related to Intenet addiction(r=0.06, 0.07, 0.27, 0.44, 0.07, 0.11, 0.09, P<0.01), while medium and high intensity sports, effective physical education, health education in school, AIDS education and parental restrictions on TV and computer games, were negatively associated with internet addiction(r=-0.74, -0.65, -0.04, -0.05, -0.63, P<0.01). Conclusion The internet addiction of Chinese primary school students is common and increases by year. High-intensity, high-density sports, effective physical education, and health education could help prevent Internet addiction among children.

Objective To investigate the efficacy and the collective brain responses of acupuncture for post-stroke limb numbness.Methods A total of 24 patients with post-stroke limb numbness and 23 matched healthy participants were recruited.Both groups received coordinated acupuncture treatment to regulate their physical and mental state,three times a week for 4 weeks.Changes in limb numbness assessed by the visual analog scale(VAS)were observed before and after acupuncture.Functional magnetic resonance imaging(fMRI)scans were conducted on the participants during the scanning process with acupuncture intervention.Blood oxygen level-dependent(BOLD)data were collected in three states:resting state,acupuncture task state,and needle retention state.Data processing was performed using software such as fMRIPrep and brainIAK.Intersubject correlation(ISC)analysis was conducted to compare the ISC values of different brain regions between the two groups under different conditions.Results Acupuncture treatment significantly reduced the VAS scores of limb numbness.No significant changes in ISC values in both groups during resting state.Under acupuncture conditions,the patient group exhibited significantly increased ISC values in the left precentral gyrus,left paracentral lobule,left postcentral gyrus,left insula,left anterior cingulate gyrus,right middle frontal gyruss and right anterior cingulate gyrus.Under the needle retention condition,a slight negative activation was observed in the bilateral precentral gyrus of the control group.In the patient group,the ISC values of the left superior frontal gyrus,left precentral gyrus,left paracentral lobule,left precuneus,left postcentral gyrus,left insula,left anterior cingulate gyrus,left thalamus,right superior frontal gyrus,right middle frontal gyrus,right precentral gyrus,and right anterior cingulate gyrus showed a significant increase compared to the resting state.Conclusion Acupuncture can improve post-stroke limb numbness symptoms and induce a specific pattern of collective brain activation in patients.This activation involves the precentral gyrus,postcentral gyrus,thalamus,insula,frontal lobe and cingulate gyrus.Acupuncture may enhance the processing of sensory information by activating the somatosensory-motor network and the cortical-thalamic-cortical loop.It may also modulate the central executive network and the descending pain control pathway to promote the relief of pain symptoms.

Objective To study the effect of amino acid site variation of HIV-1 Nef protein on its inhibition of neuronal autophagy and explore the mechanism of Nef protein-induced central nervous system injury.Methods HIV-1 nef genes were amplified and cloned from the temporal cortex (TC) of the central nervous system in 1 case of HIV-associated dementia (HAD) H and 1 case of non-HAD AIDS patient N.The amino acid sequences were aligned by NCBI BLAST tools and MEGA6.0 software to study the variation of amino acid sites.The eukaryotic expression vectors pEGFP-N1-nef derived from H-TC and N-TC were constructed and transfected into U87 cells to observe green fluorescence.At the same time,the expression of Nef protein,LC3-Ⅱ and p62 protein in U87 cells were detected by Western blot,and the effects of different sources of Nef on autophagy of U87 cells were analyzed.Results The nef genes were amplified by PCR and clone vectors pMD-19T-nef of H-TC and N-TC were successfully constructed.The sequencing confirmed that they were HIV-1B subtypes.The amino acid sequence analysis showed that there were differences between H-TC and N-TC key amino acid sites.The recombinant plasmid pEGFP-N1-nefwas successfully constructed and expressed Nef protein in U87 cells.Western blot analysis showed that the expression of LC3-Ⅱ protein was significantly different among groups (F =11.764,P =0.001).There was no significant difference in the expression of LC3-Ⅱ between cell control and plasmid control(P=0.169).The content of LC3-Ⅱ was low in the two groups of cells,which could not be detected by Western blot.The expression of LC3-Ⅱ in H-TC,N-TC and positive control CQ group increased,compared with blank control or blank vector.The difference was statistically significant (P=0.017,P=0.039,P=0.031),and the expression of LC3-Ⅱ in H-TC group was higher than that in N-TC group (P =0.023);the expression of p62 protein in H-TC,N-TC and positive control CQ group was higher than that in blank control or blank vector group,but there was no significant difference between groups (F=2.049,P =0.163).Conclusions HIV-1 Nef amino acid sites in the central nervous system of patients with HAD and non-HAD were different,and their effects on autophagy of U87 cells were different.The expression of LC3-Ⅱ,an autophagic marker protein,was more strongly induced by Nef from H-TC.

Objective: To study the sequence characteristics and variation of HIV-1 Vpr gene in different parts of an AIDS dementia complex (ADC) patient and provide basis for the study of the neurologic pathogenesis of HIV-1-assciatd dementia. Methods: Genomic DNA was extracted from peripheral samples (lymph nodes, spleen, liver) and central nervous system (meninges, frontal lobe, temporal lobe gray matter, frontal white matter, basal ganglia cortex) of an ADC patient, The Vpr gene was amplified with nested polymerase chain reaction (PCR). PCR products were cloned into the pMD19-T vector. After transformation into DH5α competent E. coli, five positive clones were sequenced. The phylogenetic tree was built and genetic distance was calculated through MEGA6, and the values of ds/dn was calculated through SNAP, then the changes of the amino acid sites were analyzed. Results: HIV-1 Vpr genes isolated from different tissues of the ADC patient had variations. Vpr HIV-1 gene sequences from central nervous system and peripheral tissues were intercrossed together in the phylogenetic tree. Central nervous system and peripheral HXB2 Vpr had no significant differences in genetic distance. The ds/dn of all the HIV-1 Vpr gene sequences were 3.3749. Conclusions The HIV-1 Vpr sequences were different in the ADC patient, and there were different variations in different parts of the peripheral and central regions. Whether these variations are related to the pathogenesis of ADC remains to be further studied.

Objective: BG-derived HIV-1 Tat protein from an HIV-associated dementia (HAD) patient was expressed in E. coli BL21(DE3) and purified in order to research the effects on human umbilical vein endothelial cells (HUVECs) activity. Methods: The recombinant plasmid pGEX-KG-tat with HIV-1 tat stored in our laboratory was amplified by PCR. The PCR product was cloned into pET-32a-tat. The recombinant plasmid pET-32a-tat was transfected into E. coli, and Tat protein was expressed in BL21(DE3), which was induced by IPTG. Then it was purified by Ni-chelating chromatography column and gel filtration preloaded column, and identified by SDS-PAGE and Western blot(WB). The concentration was determined by BCA Kit. Different concentrations of Tat were added into HUVECs to detect their effects on cell activity by cck-8. Results: The Tat with high purity was efficiently expressed in BL21 (DE3) and obtained by using the Ni-chelating chromatography column and gel filtration preloaded column. The concentration was 0.47 mg/ml by using BCA Kit. As the concentration of Tat increased, HUVECs activity decreased. There was no significant difference in cells viability between negative control with 100 ng/ml and 200 ng/ml group (P>0.05). There was significant difference in cells viability between negative control with 300 ng/ml, 400 ng/ml, 500 ng/ml and 1000 ng/ml group (P<0.05). But the difference between 300 ng/ml, 400 ng/ml, 500 ng/ml and 1000 ng/ml group was not statistically significant (P>0.05). Conclusions The HIV-1 Tat with biological activity was efficiently expressed in BL21 (DE3), and the activity of HUVECs was significantly decreased when the concentration reached 300 ng/ml.

Enzalutamide (ENZ) is a second-generation androgen receptor (AR) antagonist used for the treatment of castration-resistant prostate cancer (CRPC) and reportedly prolongs survival time within a year of starting therapy. However, CRPC patients can develop ENZ resistance (ENZR), mainly driven by abnormal reactivation of AR signaling, involving increased expression of the full-length AR (ARfl) or dominantly active androgen receptor splice variant 7 (ARv7) and ARfl/ARv7 heterodimers. There is currently no efficient treatment for ENZR in CRPC. Herein, a small molecule LLU-206 was rationally designed based on the ENZ structure and exhibited potent inhibition of both ARfl and constitutively active ARv7 to inhibit PCa proliferation and suppress ENZR in CRPC. Mechanically, LLU-206 promoted ARfl/ARv7 protein degradation and decreased ARfl/ARv7 heterodimers through mouse double minute 2-mediated ubiquitination. Finally, LLU-206 exhibited favorable pharmacokinetic properties with poor permeability across the blood-brain barrier, leading to a lower prevalence of adverse effects, including seizure and neurotoxicity, than ENZ-based therapies. In a nutshell, our findings demonstrated that LLU-206 could effectively inhibit ARfl/ARv7-driven CRPC by dual-targeting of ARfl/ARv7 heterodimers and protein degradation, providing new insights for the design of new-generation AR inhibitors to overcome ARfl/ARv7-driven CRPC.