Objective To evaluate the safety of the low glucoside composites of Epimedii Folium and clarify the pharmacokinetic characteristics of its five low glucosides.Methods Four groups of KM mice were orally administrated of corn oil,1 968,2 625 and 3 500 mg·kg-1 low glucoside composites of Epimedii Folium,respectively.Then,the living conditions,toxic symptoms,and death of the mice were observed for 7 consecutive days.After the mice were dissected,the viscera/body ratio and the viscera/brain ratio were calculated.Besides,the contents of alanine aminotransferase(ALT)and aspartate transaminase(AST)in plasma were determined by ELISA,and the pathological changes of the liver were observed by HE staining.C57BL/6J mice were intravenously or orally administered of baohuoside I,baohuoside II,sagittatoside A,sagittatoside B and sagittatoside C.Then,blood samples were collected at different time points.The plasma concentrations of the five low glucosides were measured by UHPLC-MS/MS.Results When compared with the control group,no significant differences were found in the body mass,viscera/body ratio,viscera/brain ratio,contents of ALT and AST in plasma after oral administration of different doses of low glucoside composites to mice.Moreover,no pathological changes or damages were found in the liver sections.After intravenous injection,the AUC0-t values of baohuoside Ⅰ,baohuoside Ⅱ,sagittatoside A,sagittatoside B and sagittatoside C in mice were 4.82,82.54,276.64,88.77 and 178.02 min·μg·mL-1,respectively.Meanwhile,the t1/2 values were 60.42,115.27,67.63,131.61 and 129.87 min,respectively.After oral administration,the AUC0-t values of the five low glucosides were 31.64,18.59,3.48,2.41 and 2.42 min·μg·mL-1,respectively.The Cmax values were 147.23,86.76,15.58,24.34 and 26.12 ng·mL-1,respectively.The tmax values were 21.00,78.00,78.00,30.00 and 28.00 min,respectively.The bioavailability of baohuosideⅠ,baohuosideⅡ,sagittatoside A sagittatoside B and sagittatoside C were 1.91%,0.51%,0.05%,0.06%and 0.04%,respectively.Conclusion The low glucoside composites of Epimedii Folium has high safety,and no hepatotoxicity were observed at dose of 3 500 mg·kg-1.The 5 low glucosides are quickly absorbed and rapidly eliminated in mice,and all of them have low bioavailability.

Objective To systematically evaluate the efficacy and safety of nivolumab in the treatment of non-small cell lung cancer.Methods PubMed,Embase,Cochrane Library,China National Knowledge Infrastructure(CNKI),Weipu Chinese Science and Technology Journal Database,Wanfang Medical Database were searched for articles published from the establishment of the database to March 2023.Published randomized controlled clirical trials of nivolumab in the treatment of patients with non-small cell lung cancer were selected,overall survival,progression-free survival,and adverse reaction rate as outcome indicators were used.A meta-analysis using STATA version 13.1 statistical software was conducted.Results A total of 8 phase Ⅲrandomized controlled trials involving 4 945 subjects were included.Compared with the traditional chemotherapy group,patients in the nivolumab group had significantly reduced risk of death in terms of overall survival(HR=0.73,95%CI=0.65-0.82,P<0.05),and in terms of progression-free survival,nivolumab significantly reduced the risk of recurrence compared with the traditional chemotherapy group(HR=0.74,95%CI=0.63-0.88,P<0.05).In terms of safety,there was no significant difference between the nivolumab group and the traditional chemotherapy group for diarrhea,but the incidence of nausea,neutropenia,anemia,decreased appetite,and fatigue in the nivolumab group was lower than that in the traditional chemotherapy group.However,it should be worth noting that the incidence of immune-related adverse events such as rash was higher in the nivolumab group than in the traditional chemotherapy group,and the difference was statistically significant(OR=3.85,95%CI=2.05-6.25,P<0.05).Conclusion Compared to traditional chemotherapy,the efficacy and safety of nivolumab in the treatment of non-small cell lung cancer were better,but the risk of immune-related adverse events increased.

Objective To investigate the effect of combination of rope-assisted proprioceptive neuromuscular facilitation(PNF)training and rope-assisted brain-computer interface(BCI)training on upper limb function in stroke patients with hemiplegia. Methods From March,2022 to February,2023,96 inpatients with stroke hemiplegia from the Second Affiliated Hospital of Guangxi Medical University were randomly divided into conventional group(n=32),PNF group(n=32)and combined group(n=32).All the groups received routine rehabilitation treatment.The conventional group re-ceived upper limb PNF training,the PNF group received upper limb rope-assisted PNF training,and the com-bined group received both upper limb rope-assisted PNF training and upper limb rope-assisted BCI training,for four weeks.They were assessed with Functiongal Test for the Hemiplegic Upper Extremity-Hong Kong version(FTHUE-HK),Fugl-Meyer Assessment-Upper Extremities(FMA-UE)and modified Barthel Index(MBI)before and after treatment. Results The intra-group effect(F>341.219,P<0.001),inter-group effect(F>21.705,P<0.001)and interaction effect(F>3.171,P<0.05)were significant in the scores of FTHUE-HK and MBI.The intra-group effect(F=520.472,P<0.001)and inter-group effect(F=41.939,P<0.001)were significant in the scores of FMA-UE,and the interaction effect was not(P>0.05).After treatment,the FTHUE-HK,FMA-UE and MBI scores were the best in the combined group(P<0.05). Conclusion The combination of rope-assisted PNF training with rope-assisted BCI device training could further improve the motor function of the upper limbs in stroke patients with hemiplegia,and enhance their activities of daily liv-ing.

OBJECTIVE To observe the auxiliary analgesic effect of electroacupuncture at Hegu and Neiguan combined with in-domethacin suppository in patients undergoing transvaginal ultrasound-guided oocyte retrieval(TUGOR and its effect on the outcome of in vitro fertilization(IVF.METHODS 64 IVF-ET patients undergoing TUGOR were randomly divided into a treatment group and a control group,32 cases in each group.One case dropped out of the treatment group during the treatment.The control group was given indomethacin suppository rectal administration,and the treatment group was given electroacupuncture at Hegu and Neiguan in addition to the treatment of the control group.The patients'tenderness threshold,VAS score,pain grade score,respiratory rate,and pulse rate were evaluated before and after TUGOR operation.The number of oocytes obtained,the rate of two pronuclei(2PN,embryo utiliza-tion rate,and high-quality embryo rate were evaluated after TUGOR operation.The adverse reactions of the two groups were monitored during and after operation.RESULTS After TUGOR operation,the VAS score and pain grade of the treatment group were signifi-cantly lower than those of the control group(P<0.01;the tenderness threshold of the two groups was significantly reduced after opera-tion(P<0.05,P<0.01,and the treatment group was better than the control group(P<0.05;the incidence of nausea during opera-tion,abdominal distension and nausea 48 h after operation in the treatment group were better than those in the control group(P<0.05;the high-quality embryo rate in the treatment group was better than that in the control group(P<0.05.CONCLUSION Electroacupuncture at Hegu and Neiguan combined with indomethacin suppository can effectively assist in analgesia,and can reduce the incidence of adverse reactions during and after TUGOR to varying degrees,and may have certain advantages in improving the rate of high-quality embryos.

Objective:To establish a general method for the simultaneous determination of hydroxyphenyl esters and quaternary ammonium bacteriostatic agents in eye drops.Methods:The chromatographic analysis was per-formed on an Agilent C18 column(4.6 mm ×250 mm,5 μm)with 1％triethylamine solution(pH adjusted to 5.0 with phosphoric acid)as mobile phase A and methanol as mobile phase B.Gradient elution was performed at col-umn temperature of 40 ℃.The detection wavelength was 214 nm,the flow rate was 1 mL·min-1,and the injec-tion volume was 20 μL.Results:Methylparaben,ethylparaben,propylparaben,butylparaben,benzalkonium chlo-ride and benzalkonium bromide were 0.11-559.0,0.10-513.0,0.10-258.8,0.11-270.5,1.07-537.0 and 1.03-512.8 μg·mL-1,respectively.The linear range was good(r＞0.999).The average recoveries of meth-ylparaben,benzalkonium bromide and benzalkonium chloride were 104.7％(RSD=1.3％),102.6％(RSD=1.1％)and 100.9％(RSD=1.1％),respectively.The contents of bacteriostatic agent in 100 batches of eye drops from 36 varieties of 12 enterprises were determined,and the accurate results were obtained.Conclusion:This meth-od provides a reference for the content quality control and safety evaluation of bacteriostatic agents in eye drops.

Objective:To discuss the inhibitory effect of Schisandrin B on the proliferation of pancreatic cancer Pan02 cells,and to clarify the mechanism.Methods:CCK-8 method was used to detect the proliferation rates of the Pan02 cells after treated with different concentrations(0,0.78,1.56,3.12,6.25,12.50,and 25.00 mg·L-1)of Schisandrin B to select the optimal concentration and treatment time of Schisandrin B.The mouse pancreatic cancer Pan02 cells were divided into control group(0 mg·L-1 Schisandrin B),2.5 mg·L-1 Schisandrin B group,5.0 mg·L-1 Schisandrin B group,and 10.0 mg·L-1 Schisandrin B group.The morpholoy of Pan02 cells invarious groups was observed with light microscope;5-ethynyl-2'-deoxyuridine(EdU)staining assay was used to detect the positive expression rates of the Pan02 cells in various groups;flow cytometry was used to detect the percentages of the Pan02 cells at different cell cycles and the apoptotic rates of the cells in various groups;Western blotting method was used to detect the expression levels of cell cycle and apoptosis-related proteins in the cells in various groups.Results:The CCK-8 method results showed that after treated with Schisandrin B for 48 and 72 h,compared with 0 mg·L-1 Schisandrin B,the proliferation rates of the Pan02 cells after treated with different concentrations of Schisandrin B were decreased(P<0.01),especially at 72 h.0.25,5.0,and 10.0 mg·L-1 Schisandrin B were selected to treat the Pan02 cells,and 72 h was the treatment time.In control group,the Pan02 cells had a spindle shape,with good condition,and grew closely adhered to the wall with normal organelles and cytoplasm,in 2.5 and 5.0 mg·L-1 Schisandrin B groups,the cell volume was decreased,the intercellular adhesion was disappeared,and the cell membrane was intact but more permeable;the cytoplasm shrank and vacuolar structures appeared inside the cells,with some fragmented and floating on the surface of the solution;in 10.0 mg·L-1 Schisandrin B group,the Pan02 cells exhibited notable apoptotic bodies,indicating an apoptotic state.The EdU staining results showed that compared with control group,the rates of EdU positive cells in 2.5,5.0,and 10.0 mg·L-1 Schisandrin B groups were significantly decreased(P<0.01).The flow cytometry results showed that compared with control group,the percentages of the cells at S phase in 2.5,5.0,and 10.0 mg·L-1 Schisandrin B groups were significantly increased(P<0.01),while the percentages of the cells at G2/M phase were significantly decreased(P<0.01),and the percentages of the cells at G0/G1 phase in 5.0 amd 1.0 mg·L-1 Schisandrin groups were decreased(P<0.01);compared with control group,the apoptotic rates of the cells in 2.5,5.0,and 10.0 mg·L-1 Schisandrin B groups were significantly increased(P<0.01).The Western blotting results showed that compared with control group,the expression levels of p27,B-cell lymphoma 2(Bcl-2)associated X protein(Bax),cleaved cysteine aspartic acid protease-3(cleaved Caspase-3),and cleaved poly adenosine diphosphate(ADP)ribose polymerase(cleaved PARP)proteins in the cells in 2.5 mg·L-1 Schisandrin B group were significantly increased(P<0.05 or P<0.01),the expression levels of cyclin A2,cyclin E2,and Bcl-2 proteins in the cells in 5.0 and 10.0 mg·L-1 Schisandrin B groups were significantly decreased(P<0.05 or P<0.01),while the expression levels of p27,Bax,cleaved Caspase-3,and cleaved PARP proteins in the cells in 5.0 and 10.0 mg·L-1 Schisandrin B groups were significantly increased(P<0.01).Conclusion:Schisandrin B has an inhibitory effect on proliferation of the pancreatic cancer Pan02 cells,and its mechanism may be related to the activation of the cysteine aspartic acid protease-3(Caspase-3)pathway to induce the apoptosis and activating p27 protein to induce the arrest of cell cycle at S phase.

Objective To establish human and mouse pancreatic cancer cell strains stably expressing Cas9 protein,green fluorescent protein,red fluorescent proteins,luciferase-tdTomato,and to validate the activity of luciferase and gene editing of Cas9 function for pancreatic cancer research using luciferase and CRISPR/Cas9 system.Methods In human pancreatic cancer cells(AsPC-1,CFPAC-1,HPAC,BxPC-3,HS 766T,MIA PaCa-2,PANC-1,and SW 1990),and mouse pancreatic cancer cell(Pan02),the cells were infected with Cas9-expressing plas-mid pLv-EF1α-Cas9m1.1-Puro,and single-cell clones were selected for culture and expansion.After extracting the total protein,Western blot verified the expression level of Cas9;Infected with fluorescent protein expression plasmids pLv-EF1α-EGFP,pLv-EF1α-mCherry,pLv-EF1α-tdTomato,pLv-EF1α-Luc2-tdT,and selected single cell clones stably expressing fluorescent proteins were cultured and amplified under fluorescence microscope.Cas9 stable expression cell line was selected to be infected with pLv-EF1α-Luc2-tdT,and the mono-clonal culture of stable expression of fluorescent proteins was selected for expansion under fluorescence micro-scope.One of the cell lines were selected to be infected with Lv-EF1a-mCherry,and the mCherry-positive cells were sorted out by flow cytometry,and then the guide RNA of mCherry gene was then infected by lentivirus to tar-get the mCherry gene,and after cell expansion,mCherry knockdown was detected by fluorescence microscope observation and flow cytometry;5 BALB/c Nude mice were subcutaneously inoculated with MIA PaCa-2-Luc2-tdT cells(1.0×107/cells each),and imaged in vivo after 36 days.Results 48 human pancreatic cancer cell strains with stable Cas9 expression were screened(including 23 cells expressing Cas9m1.1,25 cells expressing Cas9m1.1-Luc2-tdT),33 pancreatic cancer cell strains with stable expression of fluorescent proteins were screened(8 cells expressing EGFP,7 expressing mCherry,and 9 each expressing Luc2-tdT and tdTomato).Cells expressing mCherry and Cas9 were infected with mCherry gRNA and mCherry was knocked down.In vivo imaging showed that both bioluminescence and fluorescence luminescence were present in MIA PaCa-2 cells ex-pressing Luc2-tdT.Conclusions 33 pancreatic cancer cell strains with stable expression of fluorescent proteins are successfully established,in which the Luc2-tdT-expressing cell strains have luciferase activity;48 pancreatic cancer cell strains with stable expression of Cas9 are successfully established,and the Cas9 protein has gene edi-ting activity,gene editing activity varies depending on the original cell strains.

Visual hallucination(VH)is a common symptom of schizophrenia,the underlying mechanism has not been fully elucidated.It has been found that the dysfunction of dopamine(DA)system,the overactivation of α-amino-3-hydroxy-5-methyl-4-isoxazole-propionate(AMPA)receptor in glutamate system and the dysfunction of γ-aminobutyric acid(GABA)ergic neurons can induce VH in patients with schizophrenia.In addition,abnormalities in brain structural and functional networks and visual networks are also closely related to the occurrence of VH.The purpose of this paper is to review the neurochemistry and nerve injury mechanism of VH in schizophrenic patients to deeply understand the characteristics of VH,and make more accurate judgment in the early diagnosis,condition evaluation and treatment plan of schizophrenic patients.

Peritoneal adhesions are one of the most common postoperative complications.The formation of peritoneal adhesions is a complex process with multiple factors and stages.Various inflammatory cells and their secreted cytokines promote the chronicity of inflammatory response,the initiation of coagulation cascade reaction and excessive deposition of fibrin,ultimately leading to the formation of pathological peritoneal adhesions.Research in recent years has also highlighted the important role of non-coding RNA in peritoneal adhesions.

ObjectiveTo investigate the effect and mechanism of Shenling Baizhusan on the treatment of oligoasthenospermia with hyperuricemia （HUA）. MethodThirty-two male Kunming （KM） mice were randomly divided into blank group （n=6）， model group （n=6）， high-dose Shenling Baizhusan group （n=7）， low-dose Shenling Baizhusan group （n=7）， and febuxostat group （n=6）. Except for the blank group， all other groups received intraperitoneal injection of potassium oxazinate suspension （600 mg·kg^-1） for 7 days. After modeling， the high-dose Shenling Baizhusan group and the low-dose Shenling Baizhusan group were orally administered with 20.14 g·kg^-1 and 10.07 g·kg^-1 of Shenling Baizhusan， respectively. The Febuxostat group was orally administered with 0.25 g·kg^-1 of Febuxostat， while the blank group and model group were orally administered with the same volume of physiological saline. Oral administration was performed once a day for 14 consecutive days， after which samples were collected. Biochemical methods were used to measure serum uric acid （UA）， superoxide dismutase （SOD） and malondialdehyde （MDA） in testicular tissue. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes in testicular tissue and evaluate the spermatogenesis function. Automated sperm analyzer was used to measure sperm density and motility. Single-cell gel electrophoresis （SCGE） was used to assess sperm DNA integrity. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） was used to detect testicular cell apoptosis rate. Western blot analysis was performed to measure the protein expression levels of Kelch-like ECH-associated protein 1 （Keap1）， nuclear factor E₂-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and Caspase-3 in testicular tissue. Real-time polymerase chain reaction （PCR） was conducted to evaluate the mRNA expression levels of Keap1， Nrf2， and HO-1 in testicular tissue. ResultCompared with the blank group， the model group showed elevated serum UA level （P<0.01）， decreased testicular spermatogenesis function， sperm density， and motility （P<0.01）， and increased sperm trailing rate and testicular cell apoptosis rate （P<0.01）. Compared with the model group， the high-dose Shenling Baizhusan group showed significant improvements in the above-mentioned indicators （P<0.05， P<0.01）. Additionally， the expression levels of Keap1， Bax， and Caspase-3 in testicular tissue were reduced， while the expression levels of Nrf2， HO-1， and Bcl-2 increased （P<0.05， P<0.01）. The mRNA level of Keap1 decreased （P<0.05， P<0.01）， while the mRNA levels of Nrf2 and HO-1 increased （P<0.05， P<0.01）. ConclusionShenling Baizhusan can significantly improve HUA oligoasthenospermia， and its mechanism may be related to the Nrf2/antioxidant response element （ARE） signaling pathway.