BACKGROUND:Previous studies have shown that puerarin can inhibit the differentiation of osteoclasts,and the expression of Notch signaling pathway-related proteins such as Notch1,HES1,and Jagged1 is decreased.However,the specific mechanism of the Notch1 signaling pathway for the inhibition of osteoclast differentiation by puerarin is not clear. OBJECTIVE:To explore the effect of Notch signaling pathway on puerarin inhibiting the differentiation of mouse macrophage Raw264.7 into osteoclasts. METHODS:Raw264.7 cells were divided into seven groups for intervention culture.Blank control group was cultured in high-sugar DMEM medium;the osteoclast induction group was cultured in osteoclast induction medium;the puerarin intervention group was cultured with 50 μmol/L puerarin at the same time of osteoclast induction;Notch1 siRNA control group,Notch1 siRNA group,Notch1 overexpression control group and Notch1 overexpression group were transfected with Notch1 siRNA control sequence,Notch1 siRNA,Notch1 overexpression control plasmid and Notch1 overexpression plasmid,respectively,and then cultured with osteoclast induction medium and puerarin.The number and size of osteoclasts were observed by tartrate-resistant acid phosphatase staining,the skeleton formation of osteoclasts was observed by F-actin staining,and the gene expression level of osteoclast formation markers was detected by RT-PCR. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining results showed that puerarin intervention could inhibit the generation of osteoclasts,Notch1 silencing could further reduce the number of osteoclasts,while the number of osteoclasts in the osteoclast-induced group increased significantly after Notch1 overexpression.The results of F-actin showed that Raw264.7 cells could form a well-defined F-actin ring after osteoclast induction.Puerarin intervention would inhibit the formation of cytoskeleton,and Notch1 silencing could aggravate the inhibitory effect of cytoskeleton formation,while Notch1 overexpression could alleviate this inhibitory effect of puerarin.RT-PCR results showed that puerarin could inhibit the mRNA expression levels of tartrate-resistant acid phosphatase,Cathepsin K and c-Fos,the expression of the above-mentioned three factors decreased significantly after Notch1 gene silencing,and Notch1 overexpression could upregulate the expression of these factors.These finding indicate that puerarin inhibits the differentiation of Raw264.7 cells into osteoclasts through the Notch signaling pathway.

In recent years， the research data on the combination model of insomnia of liver depression and qi stagnation syndrome in traditional Chinese medicine （TCM） were sorted out and summarized in terms of the selection of experimental animals， modelling methods， and model evaluation， with a view to providing references for the standardized preparation of the experimental model of insomnia of liver depression and qi stagnation syndrome and the study of the intervention mechanism. As for the selection of experimental animals， rats and mice are commonly used for the study of sleep patterns in the insomnia model of liver depression and qi stagnation syndrome because their sleep patterns are similar to those of human beings； zebrafish is suitable for the study of sleep patterns in early development because of its high genetic homology； and rhesus macaques， common marmosets， and crab-eating monkeys are used for the study of highly complex sleep disorders because their physiological and behavioural characteristics are closer to those of human beings. In terms of modelling methods， electrical stimulation， chronic restraint， chronic emotional stress and impact psychological stress each have their own characteristics and application scenarios， while chronic tail-clamping stimulation combined with intraperitoneal injection of p-chlorophenylalanine （PCPA） combines physical and chemical stimuli， in order to simulate the complex mechanism of insomnia in human beings， and to more comprehensively simulate the pathology of insomnia with liver depression and qi stagnation syndrome. In terms of evaluation indexes， electroencephalography （EEG） and electromyography （EMG） monitoring and barbiturate synergistic sleep test are the most commonly used and scientific methods for evaluating insomnia animal models with liver depression and qi stagnation syndrome， which can provide intuitive and continuous sleep state monitoring； barbiturate synergistic sleep test can reflect the sensitivity and responsiveness of the model animals to the sleep-regulating medications； and the introduction of the method of using formulas to measure syndromes for the evaluating the model can combine the treatment and diagnostic principles of TCM with modern scientific research.

ObjectiveTo explore the efficacy of acupuncture intervention for children with monocular refractive-parallax amblyopia and the possible mechanisms of brain function based on resting-state functional magnetic resonance （rs-fMRI）. MethodsSeventy-six children with anisometropic amblyopia were randomly divided into routine treatment group （38 cases） and acupuncture treatment group （38 cases）. In the conventional group， the children were given three regular treatments of red flash， grating and visual stimulation for 5 mins each time； in the acupuncture group， on the basis of the conventional treatment， the children were given acupuncture for 20 mins each time on bilateral Jingming （BL 1）， Cuanzhu （BL 2）， Guangming （GB 37） and Fengchi （GB 20）； the children in both groups were treated once every other day and three times a week for 4 weeks. The corrected visual acuity was compared between groups before and after treatment. Fifteen children with left-sided refractive amblyopia were randomly selected from each of the above two groups and underwent brain rs-fMRI scans before and after treatment， and 10 healthy children with normal visual acuity of the matched gender and age were included in the normal group and underwent brain rs-fMRI scans. Based on the activation likelihood estimation （ALE） method， we constructed the what visual pathway network， and compared and analyzed the spherical regions of interest （ROIs） of the children with normal children， and both groups of children with differences in functional connectivity （FC values） within the what pathway in the brain before and after treatment. ResultsTwo cases dropout in the acupuncture group， and finally 36 cases in the acupuncture group and 38 cases in the conventional group were included in the analysis. Compared with before treatment， the best corrected visual acuity of amblyopia in both groups was significantly improved after treatment （P＜0.05）， and the improvement of vision in the acupuncture group was significantly better than that in the conventional group （P＜0.05）. The results of rs-fMRI showed that the FC values of the primary optic cortex and the right fusiform gyrus， the left lingual gyrus and the right fusiform gyrus， and the right infraoccipital gyrus and the right middle temporal gyrus were significantly elevated in the brain of the refractive amblyopia children with the whitepathic amblyopia， compared with that of the normal children （P＜0.05）. The FC values of the left lingual gyrus， the right suboccipital gyrus with the right fusiform gyrus， the left lingual gyrus with the right middle temporal gyrus， the right and left lateral middle occipital gyrus， and the right and left lateral middle occipital gyrus with the right suboccipital gyrus were significantly （P＜0.05） lower in the conventional group compared with those in the conventional group before treatment. Compared with that before acupuncture treatment， the FC values of the right lingual gyrus and the right fusiform gyrus， the primary visual cortex and the right middle temporal gyrus increased significantly after acupuncture treatment （P＜0.05）， and the FC values of the left inferior occipital gyrus and the right middle temporal gyrus， the FC values of the left lingual gyrus and the right middle occipital gyrus decreased significantly （P＜0.05）. Compared between groups after treatment， the FC between the left suboccipital gyrus and the right fusiform gyrus in the acupuncture group was significantly higher than that in the conventional group （P＜0.05）， and the FC between the left middle occipital gyrus and the right and left suboccipital gyrus was significantly lower than that in the conventional group （P＜0.05）. ConclusionAcupuncture can significantly improve the corrected vision of anisometropic amblyopic children， and its effect mechanism may focus on regulating the occipito-temporal interlobular functional connectivity within the what pathway， thus improving the children's visual function of shape and color vision and visual learning and memory ability.

Objective:To observe the effects of liver-soothing spirit-regulating acupuncture on the behaviors and expression levels of 5-hydroxytryptamine receptor 1A(5-HT1AR)and 5-hydroxytryptamine receptor 2A(5-HT2AR)in the hippocampus of insomnia rats due to liver Qi stagnation. Methods:Ten of the 70 Wistar rats were randomly selected as the blank group,and the remaining rats were subjected to preparing the insomnia model due to liver Qi stagnation by the chronic tail clamping stimulation combined with intraperitoneal injection of para-chlorophenyl alanine.Fifty successful model rats were randomly divided into a model group,a grasp group,a Western medicine group,an acupuncture group,and a sham acupuncture group,with 10 rats in each group.Rats in the grasp group were grabbed in the same way as those in the acupuncture group.Rats in the Western medicine group were given estazolam solution by intragastric administration.Rats in the acupuncture group received liver-soothing spirit-regulating acupuncture at Baihui(GV20),Neiguan(PC6),Shenmen(HT7),and Taichong(LR3);rats in the sham acupuncture group received acupuncture at 4 non-meridian non-point spots.All groups were treated once a day for 7 d.Pentobarbital sleep test was used to evaluate the sleep latency(SL)and sleep time(ST)of rats in each group after 7 d of intervention;the proportion of the number of entry into the open arms(OE％)and the proportion of the residence time in the open arms(OT％)were monitored by elevated plus maze test(EPMT);the open field test(OFT)was used to monitor the numbers of standing upright,horizontal cross-grid,and fecal grains in rats.The expression levels of 5-HT1AR and 5-HT2AR in rat hippocampus were detected by Western blotting. Results:The body mass gain,ST,OE％,OT％,upright standing times,horizontal cross-grid times,and the 5-HT1AR expression level in the hippocampus were significantly decreased(P<0.01),while the SL and 5-HT2AR expression level in the hippocampus were significantly increased(P<0.01)in the model group compared with the blank group.After intervention,the body mass,ST,OT％,OE％,upright standing times,horizontal cross-grid times,and hippocampal 5-HT1AR expression level were significantly increased(P<0.01 or P<0.05),while the SL and hippocampal 5-HT2AR expression level were significantly decreased(P<0.01 or P<0.05)in the acupuncture group and the Western medicine group compared with the grasp group.Compared with the Western medicine group,OT％,SL,and hippocampal 5-HT1AR expression level were significantly increased in the acupuncture group(P<0.05).The body mass,OT％,OE％,ST,upright standing times,horizontal cross-grid times,and the 5-HT1AR expression level in the hippocampus were significantly increased(P<0.01),while the SL and 5-HT2AR expression level in the hippocampus were significantly decreased(P<0.01)in the acupuncture group compared with the sham acupuncture group. Conclusion:Liver-soothing spirit-regulating acupuncture can not only improve sleep but also improve the abnormal behaviors and mood of insomnia rats due to liver Qi stagnation,and the improvement effect on abnormal mood is better than that of Western medicine.The mechanism may be related to the regulation of 5-HT1AR and 5-HT2AR

Objective:To explore the role of blended learning in the undergraduate teaching of Clinical Biochemistry. Methods:The Batch 2017 medical laboratory technology undergraduates ( n=134) were selected as research objects, and the effect and opinions of blended learning were statistically analyzed by questionnaire survey and online-offline platform data. SPSS 23.0 was used to conduct rank sum test. Results:The application of blended learning in the Clinical Biochemistry teaching affected the learning effect in an all-round way. The average score increased from 70 (64, 76) to 79 (71, 85), with statistical difference ( Z=6.69, P<0.001). Conclusion:The combined application of blended learning, problem-based learning, flipped classroom and formative assessment is conducive to teaching students in accordance with their aptitude and cultivating students' clinical thinking ability.

Pain is a complex physiological and psychological activity, involving at least three dimensions, including pain sensation, pain emotion, and pain cognition. Acupuncture can clearly relieve the pain sensation of patients and improve pain emotion and pain cognition induced by pain; acupuncture participates in the multi-dimensional regulation of pain through brain regions of the limbic system such as anterior cingulate cortex (ACC), amygdala (AMY), and hippocampus. By analyzing relevant literature, it has been found that the regulation of acupuncture on pain emotion is mainly related to the activation of pertinent opioid receptors in the ACC, the decrease of the expression of extracellular signal-regulated kinase (ERK), and the promotion of the expression of glutamic acid (Glu) A1, metabotropic glutamate receptor-1 (mGluR1), and γ-aminobutyric acid aminobutyric acid (GABA) B2 protein in the AMY. The regulation of acupuncture on pain cognition is mainly related to the elevation of the expression of protein kinase A (PKA) and phospho-p38 mitogen-activated protein kinase (phospho-p38 MAPK) and the inhibition of cyclic adenosine monophosphate (cAMP)/PKA/cAMP response element-binding protein (CREB) signaling pathway in the ACC.

Autoantibodies ; China ; Cohort Studies ; Follow-Up Studies ; Humans ; Idiopathic Pulmonary Fibrosis

Objective:To analyze the related factors affecting the use of autogenous arteriovenous fistula (AVF), and provide a theoretical basis for prolonging the service life of AVF in hemodialysis patients.Methods:This was a retrospective study. The patients undergoing AVF and using it to maintain hemodialysis (MHD) in the First Affiliated Hospital of Nanchang University from October 2004 to June 2017 were selected as study subjects to discuss the relevant factors affecting the service life of AVF. The data of general information, dialysis and laboratory examinations were collected through questionnaire surveys, hospital case system and hemodialysis record sheets. The patients were divided into the patency group and the dysfunction group according to the status of AVF, and the related factors were compared. Multivariate Cox proportional hazard model was used to analyze the influencing factors, and Kaplan-Meier survival curve was used to determine the service life of AVF, respectively.Results:A total of 187 subjects were included in the study. The patency group had 140 cases and the dysfunction group had 47 cases. There were statistically significant differences in the proportion of diabetes, the level of serum albumin, uric acid and parathyroid hormone (PTH) between the two groups (all P<0.05). Multivariate Cox proportional hazard regression analysis showed that diabetes ( HR=9.348, 95% CI 3.507-24.918, P<0.001) and hypoalbuminemia ( HR=12.650, 95% CI 2.925-54.714, P=0.001) were risk factors for the short service life of AVF. The results of Kaplan-Meier analysis showed that the service life of AVF in patients with diabetes was significantly shorter than that in MHD patients without diabetes (Log-rank χ2=13.191, P<0.001); the service life of AVF in patients with hypoalbuminemia was significantly shorter than that without hypoalbuminemia (Log-rank χ2=13.591, P<0.001). Conclusions:Diabetes mellitus and hypoalbuminemia are risk factors for the short service life of AVF. Therefore, intervention programs should be formulated to extend the service life of AVF.

Objective:To understand the effectiveness and safety sofosbuvir/velpatasvir (SOF/VEL) combination ±ribavirin in the treatment of chronic hepatitis C virus (HCV) infection in China.Methods:A total of 96 Chinese adults with chronic HCV infection who were treated with SOF/VEL combination ± ribavirin for 12 weeks between July 2018 and February 2020 were selected. HCV RNA, routine blood test, liver, kidney and coagulation function, abdominal Color Doppler ultrasound or CT and liver stiffness were detected at baseline, 4 weeks of treatment, end of treatment and 12 weeks of follow-up. Adverse events and laboratory abnormalities during the treatment were recorded. A t-test was used to compare the measurement data between the two groups, and the analysis of variance was used for multiple group comparison.Results:A total of 93 cases (96.9%) achieved sustained virological response (SVR12), of which 3 cases had relapsed. 88 cases (91.7%, 88/96) had achieved rapid virological response (RVR). 96 cases (100%) had achieved virological response by the end of treatment (EOT). In patients with decompensated liver cirrhosis, the average baseline Child-Pugh score and Model for End-Stage Liver Disease score was 7.4±1.0, and 11.4±1.7, respectively. Among them, 12 cases of the SOF/VEL combined with RBV treatment had achieved SVR12 (100%) at 12 weeks, while only 3 of the 5 cases of single-tablet regimen of SOF/VEL had achieved SVR12 (60%). There was no significant difference between creatinine levels and baseline during or 12 weeks after treatment. The incidence of adverse events in patients with chronic hepatitis C and compensated cirrhosis was 6.3% (5/79), while that in patients with decompensated cirrhosis was 35.3% (6/17). The most common adverse events were hyperbilirubinemia, fatigue and anemia. There were no serious adverse events, deaths or discontinuation of treatment due to adverse events.Conclusion:SOF/VEL combination ± ribavirin in the treatment of various common genotypes of chronic hepatitis C, compensated cirrhosis, decompensated cirrhosis and hepatocellular carcinoma has higher SVR12 in China, and the tolerance and safety are good.