Objective:To discuss the regulatory effect of berberine(BBR)on fatty acids in the human glioma T98G cells and its effect on the cell proliferation,migration,and invasion,and to clarify its potential mechanism.Methods:The T98G cells at logarithmic growth phase were divided into control group and different concentrations(25,50,and 100 mg·L-1)of BBR groups.Cell wound healing assay was used to detect the migration rates of the cells in various groups;Transwell chamber assay was used to detect the invasion rates of the cells in various groups.The T98G cells at logarithmic growth phase were divided into control group and 100 mg·L-1 BBR group,and Mass spectrometry was used to detect the fatty acid contents in the cells in two groups.The T98G cells at logarithmic growth phase were divided into control group and different concentrations(50,100,and 150 mg·L-1)of BBR groups;Western blotting method was used to detect the expression levels of phosphatidylinositol 3-kinase(PI3K),phosphorylated PI3K(p-PI3K),protein kinase B(AKT),phosphorylated AKT(p-AKT),sterol regulatory element-binding protein 1(SREBP-1),and fatty acid synthase(FASN)in the cells in various groups.The expression of FASN was suppressed by gene silencing technology,and the T98G cells at logarithmic growth phase were divided into control group,shFASN1 group,and shFASN2 group.Western blotting method was used to detect the expression levels of FASN protein in the cells in various groups;clone formation assay was used to detect the clone formation of the cells in various groups;cell wound healing assay was used to detect the migration rates of the cells in various groups.Results:Compared with control group,the migration rates and invasion rates of the cells in different concentrations of BBR groups were decreased in a concentration-dependent manner(P<0.01),and the fatty acid content in the cells in 100 mg·L-1 BBR group was significantly decreased(P<0.01).Compared with control group,the expression levels of p-PI3K,p-AKT,SREBP-1,and FASN proteins in the cells in 150 mg·L-1 BBR group were significantly decreased(P<0.05 or P<0.01),and the expression level of SREBP-1 protein in the cells in 100 and 150 mg·L-1 BBR groups were significantly decreased(P<0.01).After suppression of FASN expression,compared with control group,the expression levels of FASN protein in the cells in shFASN1 and shFASN2 groups were significantly decreased(P<0.01),and the expression level of FASN protein in the cells in shFASN2 group was lower than that in shFASN1 group(P<0.05);compared with control group,the numbers of clone formation and migration rates of the cells in shFASN1 and shFASN2 groups were significantly decreased(P<0.01),and the migration rate of the cells in shFASN2 group was significantly lower than that in shFASN1 group(P<0.05).Conclusion:BBR interferes with fatty acid synthesis in the glioma T98G cells by reducing the expression of the PI3K/AKT/SREBP-1/FASN pathway related proteins,and decrease their migration and invasion capabilities.

Objective To compare the therapeutic effect of acupuncture at Neiguan point(PC6),acupuncture on the upper limbs and traditional medication for motion sickness.Methods Totally 30 male KM mice were randomly divided into 5 groups(n=6):blank control group,model group(acceleration exposure),acupuncture PC6 group(acupuncture at PC6+acceleration exposure),acupuncture upper limbs group(acupuncture of the arm except PC6+acceleration exposure)and traditional medicine group(difenidol administered intragastrically+acceleration exposure).The motion sickness index(MSI)and the serum levels of serotonin(5-HT),adrenaline,glucagon,insulin and blood glucose were detected.Results MSI and serum 5-HT,adrenaline,glucagon,insulin and blood glucose levels in the model group were significantly higher than those in the blank control group(P<0.05).Compared with those in the model group,MSI and 5-HT,adrenaline,glucagon and blood glucose levels were decreased in the acupuncture PC6 group and traditional medicine group(P<0.05),while insulin level was not significantly changed(P>0.05).The adrenaline level in the traditional medicine group was lower than that in the acupuncture PC6 group(P<0.05).The acupuncture upper limbs group had lower MSI and higher insulin level than the model group(P<0.05),while there were no significant differences in the levels of 5-HT,adrenaline,glucagon or blood glucose between the two groups(P>0.05).Conclusion Both difenidol and acupuncture at PC6 can effectively relieve motion sickness.

In vitro cell experiment based on serum pharmacology is a common way to study the pharmacodynamic mechanism of Chinese medicine， and screening the optimal intervention concentration of serum containing Chinese medicine is a key step in the whole experiment. Over-diluted serum containing Chinese medicine or over-high concentration leads to false negative or positive results， while the optimal concentration of medicated serum reduces the number of groups in subsequent experiments as well as the operation error. Thus it is of great significance to screen the optimal serum concentration in studying the pharmacodynamic mechanism of Chinese medicine in in vitro cell experiments. However， there has been no unified standards for the screening methods at present. After reviewing the literature in China and abroad in the past 20 years， this paper conducted an analysis from three aspects of intragastric dose of Chinese medicine， blood collection time and serum dilution degree， and then summarized the screening methods for optimal concentration of serum containing drugs， providing guidance for the study of pharmacodynamic mechanism of Chinese medicine. The screening methods mainly included "same intragastric dose， same blood collection time， and different concentrations" "different intragastric doses， same blood collection time， and same concentration" "same intragastric dose， different blood collection time， and same concentration" "different intragastric doses， different blood collection time， and different concentrations" "serum lyophilized powder" "simulation of gradient concentration of serum containing western medicine" and "pure serum intervention". Among them， the former two were the dominant ways in current related research. The above screening methods had their own advantages and disadvantages， and researchers should make a reasonable choice according to the specific requirements and actual situation of the experiments.

Objective:To identify differentially expressed genes (DEGs) associated with the progression of synovitis in RA by using bioinformatics analysis and explore the effects of DMARDs such as methotrexate, tocilizumab and rituximab on the DEGs in RA synovium.Methods:RA expression profile microarray data GSE7307、GSE12021、GSE55457、GSE55235、GSE77298、GSE89408 were acquired from the public gene chip database (GEO), including 113 synovial tissue samples from RA and 70 healthy controls (HC). At the same time, synovial expression microarrays GSE45867, GSE24742 and GSE97165 after DMARDs treatment were obtained. These data included 8 samples treated with methotrexate, 12 treated with tocilizumab, 12 treated with rituximab and 19 treated with combined tDMARDs. R software was used to screen DEGs and Venn plots using gene ontology function enrichment and Kyoto encyclopedia of genes and genomes pathway enrichment analysis. Hub genes were selected by STRING online analysis tool and Cytoscape software.Results:Compared with HC, 797 DEGs were up-regulated and 434 DEGs were down-regulated in the synovial tissue of RA. These DEGs were mainly enriched in T cell activation, immune response-activating cell surface receptor signaling pathway. Using Cytoscape and cytoHubba to obtain 5 sets of DEGs based on the STRING database model, the degree algorithm screened out 10 hub genes: LCK, SYK, PTPRC, HLA-DRA, LYN, NCAPG, TOP2A, JUN, CXCR4, CCNB1. Methotrexate treatment significantly up-regulated 20 DEGs and down-regulated 30 DEGs. Rituximab treatment up-regulated 100 DEGs and down-regulated 55 DEGs. Tocilizumab treatment up-regulated 91 DEGs and down-regulated 317 DEGs. These altered DEGs were enriched in regulating cell adhesion, leukocyte-cell adhesion, leukocyte transfer, and insulin-like growth factor receptor signaling pathways. It was worth noting that after treatment, a total of 306 high-expressing DEGs were down-regulated, and 36 low-expressing DEGs were up-regulated.Conclusion:LCK, insulin-like growth factor receptor signaling pathway, etc. are the responsible molecular mechanisms and key pivot genes for the occurrence and development of RA, and the treatment of DMARDs, which are closely related to the response of RA to the treatment of DMARDs.

Traditional Chinese Medicine (TCM) exerts anti-tumor effects by intervening in A549 cells of human lung adenocarcinoma, mainly including activating or inhibiting downstream target proteins of Bcl-2 and Bax, or forming RIP1/RIP3/MLKL complex bodies by affecting pathways such as PI3K/Akt, thereby inducing apoptosis in A549 cells of lung adenocarcinoma; blocks the cell growth phase, thereby inhibiting the proliferation of lung adenocarcinoma A549 cells; inhibits invasion and metastasis of A549 cells by affecting the MMPs pathway, STAT3 pathway, and regulating epithelial mesenchymal transition related factors; suppresses or activates the expression of related proteins or affect related signaling pathways, thereby reversing the resistance of lung cancer A549/DDP cell lines to cisplatin and paclitaxel.

Purpose/Significance To explore the characteristics and clinical significance of differentially expressed genes closely re-lated to HPV E6/E7 by using bioinformatics.Method/Process The cervical tissue and clinical information of cervical cancer in TCGA and GTEx of UCSC are used as the training set.The expression profile chip GSE63514 related to cervical cancer in GEO is used as the validation set.Firstly,the limma package of R software is used to screen DEGs of tumor and normal samples,and Venn map of genes re-lated to E6/E7 protein in MigDB is made.Survival analysis is performed by survival kit and verified by ROC and protein expression lev-els.Secondly,key genes are obtained by copy number variation and methylation correlation.Finally,the specific co-expression network is constructed and enrichment analysis and immune infiltration analysis are performed.Result/Conclusion There are 101 differentially expressed genes related to HPV E6/E7 protein,and three genes are found to have significance after screening,namely E2F1,MCM4 and PCNA.At the same time,it is found that the genes in the specific coexpression network are significantly enriched in the DNA replication and chromosome organization pathways.Immune correlation analysis shows that key genes are significantly associated with CD4 T cells,B cells and neutrophils.DNA replication,chromosome organization,etc.,are the molecular mechanisms and key genes significantly related to the development of cervical squamous cell carcinoma and HPV E6/E7 encoded proteins.

Objective:To investigate the safety and efficacy of short-segment fixation covering the fractured vertebrae via posterior intermuscular approach plus percutaneous kyphoplasty (PKP) through the outer upper edge of the base of the fractured vertebral pedicle in the treatment of osteoporotic thoracolumbar burst fracture.Methods:A retrospective case series study was used to analyze the clinical data of 56 patients with osteoporotic thoracolumbar burst fracture admitted to Ningbo No.6 Hospital from January 2018 to February 2021, including 24 males and 32 females; aged 56-72 years [(63.5±4.6)years]. All patients underwent short-segment fixation covering the fractured vertebrae via posterior intermuscular approach combined with PKP through the outer upper edge of the base of the fractured vertebral pedicle. The operation time, intraoperative blood loss, hospitalization day and surgery-related complications were recorded. The visual analogue score (VAS) of back pain, ratios of the anterior, middle and posterior height of the fractured vertebrae and kyphotic Cobb angle were compared before operation, at postoperative 2 days and at the final follow-up.Results:All patients were followed up for 12-28 months [(14.5±2.2)months]. The operation time was 55-85 minutes [(62.0±12.1)minutes], intraoperative blood loss was 80-150 ml [(94.0±18.5)ml], and hospitalization day was 5-9 days [(7.4±1.1)days]. Based on CT examination at postoperative 2 days, there were 2 patients with paravertebral cement leakage, 2 with intervertebral space leakage and 1 with intracanal leakage, but none reported associated clinical symptoms. No implant failure or fractures of adjacent segments was detected during the follow-up period. The VAS was significantly decreased from preoperative (7.5±1.2)points to (3.2±0.8)points at postoperative 2 days ( P<0.01), and the score was further lowered to (2.2±0.8)points at the final follow-up when compared with that at postoperative 2 days ( P<0.01). The ratios of the anterior, middle and posterior height of the fractured vertebrae and kyphotic Cobb angle were significantly improved at postoperative 2 days [(89.5±13.2)%, (85.8±7.9)%, (89.5±9.0)% and (5.6±3.2)°] when compared with those before operation [(48.9±11.8)%, (61.9±11.9)%, (79.9±9.8)% and (26.3±5.6)°] (all P<0.01). Slight losses were observed in the ratios of the anterior, middle and posterior height of the fractured vertebrae and kyphotic Cobb angle at the final follow-up [(87.0±12.7)%, (82.1±7.8)%, (88.6±10.0)% and (5.4±3.2)°], but not significantly different from those at postoperative 2 days (all P>0.05). Conclusion:Short-segment fixation covering the fractured vertebrae via posterior intermuscular approach plus PKP through the outer upper edge of the base of the fractured vertebral pedicle can safely and effectively treat osteoporotic thoracolumbar burst fracture, for it can significantly improve back pain, restore the height of the fractured vertebrae and correct the kyphotic deformity.

Objective:To evaluate the efficacy of scalp nerve block combined with general anesthesia in optimizing anesthesia in the patients with moyamoya disease undergoing revascularization.Methods:A total of 154 patients with moyamoya disease, aged 18-64 yr, of American Society of Anesthesiologists Physical Status classification Ⅱ or Ⅲ, undergoing elective revascularization, were divided into 2 groups ( n=77 each) using a random nunber table method: scalp nerve block combined with general anesthesia group (GN group) and general anaesthesia group (G group). Anesthesia was induced with intravenous midazolam 0.05 mg/kg, sufentanil 0.5-1.0 μg/kg, rocuronium 0.6 mg/kg and etomidate 0.2-0.3 mg/kg.After the patients were tracheally intubated after anesthesia induction, ipsilateral scalp nerve block (2 ml for supraorbital nerve block, 2 ml for supratrochlear nerve block; 3 ml for auriculotemporal nerve block, 3 ml for greater occipital nerve block, 3 ml for less occipital nerve block) was performed with 0.5% ropivacaine in GN group.The equal volume of normal saline was locally injected in G group.Anesthesia was maintained by inhalation of sevoflurane and intravenous infusion of remifentanil 0.05-0.10 μg·kg -1·min -1 and cisatracurium 0.1 mg·kg -1·h -1.The consumption of intraoperative remifentanil, requirement for postoperative rescue analgesia, nausea and vomiting, length of postoperative hospital stay, and early neurological complications were recorded.The modified Rankin Scale scores were evaluated before operation, at discharge and at 6 months after operation. Results:Compared with G group, the consumption of intraoperative remifentanil and requirement for postoperative rescue analgesia were significantly decreased ( P<0.05), and no significant change was found in the incidence of postoperative nausea and vomiting, incidence of neurological complications, length of postoperative hospital stay, and modified Rankin Scale scores at each time point in GN group ( P>0.05). Conclusions:Scalp nerve block combined with general anesthesia can increase the perioperative analgesic efficacy and is helpful in achieving a low-opioid anesthesia mode when used in the patients with moyamoya disease undergoing revascularization.

ERK pathway regulated the programmed death ligand-1 (PD-L1) expression which was linked to the response of programmed death-1 (PD-1)/PD-L1 blockade therapy. So it is deducible that ERK inhibitor could enhance the efficacy of PD-1 inhibitor in cancer immunotherapy. In this study, PD0325901, an oral potent ERK inhibitor, strongly enhanced the efficacy of PD-1 antibody

Objective To evaluate the clinical effect of postural reduction combined with miniincision screw-rod system and percutaneous kyphoplasty (PKP) in treating osteoporotic thoracolumbar burst fractures.Methods A retrospective case series study was performed for data of 35 patients with osteoporotic thoracolumbar burst fractures without neurological deficits undergone mini-incision screw-rod system fixation and PKP between January 2012 and January 2014.There were 14 males and 21 females,with a mean age of 63.2 years (range,50-72 years).Operation time,intraoperative blood loss,complications,visual analogue score (VAS),height of fractured vertebrae and kyphosis Cobb angle were recorded.Results Operation time was (49.6 ± 6.8) min,and intraoperative blood loss was (45.6 ±7.8)ml.All patients were followed up for 9-18 months (mean,13.5 months).No intraoperative or postoperative serious complications occurred,including intracanal cement leakage,breakage or loosening of the screws.VAS of back pain was decreased from (8.4 ± 1.1)points preoperatively to (3.5 ± 0.6)points postoperatively (P ＜ 0.05).Height of the fractured vertebrae was improved from (49.62％ ± 5.68)％ preoperatively to (86.64 ± 6.63) ％ postoperatively (P ＜ 0.05).Kyphosis Cobb angle was improved from (28.12 ± 1.06) °preoperatively to (5.15 ± 1.08) °postoperatively (P ＜0.05).At the final follow-up,VAS was further decrease and vertebral height and Cobb’ s showed a slight loss of correction.Conclusion Postural reduction combined with mini-incision screw-rod system and PKP can relieve back pain,restore the height of injured vertebrae,correct kyphotic deformity and reduce operation time and blood loss,indicating a minimally invasive,safe and effective procedure for treatment of osteoporotic thoracolumbar burst fractures.