Objective To optimize the microemulsion preparation of Fufang Zicao Oil(FZO).Methods The extraction conditions of microemulsion preparation of FZO were optimized by orthogonal experiment;the kinds and contents of surfactants and cosurfactants determined by pseudo-ternary phase diagram were used for the optimization of the prescription of FZO;centrifugation method was used for detecting the stability of FZO.Results The optimum extract conditions were as follows:10-mesh coarse powder of medicinal material,four times amount of solvent(sesame oil),extraction for 25 minutes.With Tween 80 and propylene glycol(7:3)as the surfactant and cosurfactant,and mixing with the oils in the proportion of 20:10,a steady microemulsion of FZO can be obtained after centrifugation by 13000 r? min-1 for 30 min.Conclusion Under the above achieved conditions,the obtained microemulsion of FZO is clear,transparent and steady.

Currently,the toxicity study of traditional Chinese medicine is faced with the following problems. Firstly,the evaluation in vitro cannot fully reflect the true state of the body. Secondly,the traditional method is not sensitive enough to the early toxicity. Lastly,the toxicity evaluation indexes cannot determine whether the compatibility of traditional Chinese medicine produces toxicity or increases toxicity systematically. The paper proposed a synthesized early evaluation research method for target organ toxicity induced by traditional Chinese medicine:screening,validation,optimization and application. This method mainly inoolves early target organ toxicity biomarkers in screening,optimi?zation,validation,biological significance explanation,and application to the traditional Chinese medicine incompatibility based on the metabolic dynamic fingerprint spectrum in order to obtain biomarkers of target organ toxicity that are sensitive and precede conventional biochemical indices for early evaluation . We attempted to analyze the pattern of chang of the biomarkers for animals acted by″18 incompatible medicaments″compatibility combination. We found that Radix Aconiti Lateralis Preparata with cardiotoxicity were compatible with Rhizoma Pinelliae,and that Trichosanthes kirilowii Maxim,Fritillaria,Ampelopsis Radix and Bletilla striata without non-cardiotoxicity produced and increased cardiotoxicity systematically.

Objective:To investigate the effects of bromodomain and extraterminal domain (BET) inhibitor JQ1 combined with siPD-L1 on the proliferation and apoptosis of oral squamous cell carcinoma (OSCC) Scc-25 cells and its mechanism.Methods:Scc-25 cells were cultured in vitro and treated with different concentrations of JQ1 (0, 0.2, 1, 5 μmol/L). Cell proliferation was detected by cell count kit-8 (CCK-8) assay; the expression levels of programmed cell death ligand 1(PD-L1) and forkhead box M1(FoxM1) protein were detected by Western blot. Appropriate concentration of JQ1 was selected for subsequent experiments. Scc-25 cells were divided into four groups: control group (without any treatment), siPD-L1 group (transfected with siPD-L1), JQ1 group (added JQ1 after transfected with non-specific siRNA), and combined treatment group (added JQ1 after transfected with siPD-L1). CCK-8 assay was used to detect the proliferation ability of Scc-25 cells in each group. Western blot was used to detect the expression levels of cleaved caspase-3, PD-L1 and FoxM1, and flow cytometry was used to detect the apoptosis rate of cells in each group. Results:With the increase of JQ1 concentration, the proliferation ability of SCC-25 cells and the expression levels of PD-L1 and FoxM1 decreased gradually (all P<0.01). JQ1 concentration of 1 μmol/L had obvious inhibitory effect on cell proliferation and the expression levels of PD-L1 and FoxM1, so JQ1 concentration of 1 μmol/L was selected for subsequent experiments. The proliferation ability of Scc-25 cells, the expression of PD-L1 and FoxM1 proteins in JQ1 group, siPD-L1 group and combination treatment group were significantly lower than those in the control group (all P<0.01), and the expression of cleaved caspase-3 protein and the rate of apoptosis were significantly higher than those in the control group (all P<0.01); Moreover, the effect of the combination treatment group was more significant than that of siPD-L1 group, JQ1 group (all P<0.01). Conclusions:The combination of JQ1 and siPD-L1 could effectively inhibit the proliferation and promotes the apoptosis of OSCC Scc-25 cells, and its mechanism may be related to the suppression of PD-L1 and FoxM1 signaling pathways.

Objective To investigate the mechanism of action of the long non-coding RNA(lncRNA)taurine up-regulating factor 1(TUG1)with high glucose(HG)-induced cellular pyroptosisin microglial cell.Methods Mouse BV2 cells were cultured and divided into normal control(NC),HG,lentiviral empty vector(sh-Con),TUG1 lentiviral gene silencing vector(sh-TUG1),HG+sh-Con and HG+sh-TUG1 group.RT-PCR was used to detect the expression and transfection efficiency of TUG1 mRNA.Nucleotide-binding oligomerized structural domain-like receptor protein 3(NLRP3),cysteoaspartate protease-1(Caspase-1),and ghrelin D(GSDMD),IL-18,IL-1β mRNA and protein expression were detected by RT-PCR and Western blot.Results TUG1 mRNA expression was higher in HG group than in NC group(P＜0.05).After transfection,a lot of green fluorescence appeared in sh-TUG1 and sh-Con group,while no green fluorescence was observed in NC group.The expression of TUG1 mRNA was lower in sh-TUG1 group than in NC group(P＜0.05).Accordingly,the recombinant lentivirus successfully infected BV2 cell.The expressions of the mRNA and protein of NLRP3,Caspase-1,GSDMD,IL-18 and L-1β were higher in HG,HG+sh-Con groups than in NC group(P＜0.05).The expressions of the mRNA and protein of NLRP3,Caspase-1,GSDMD and IL-18 and L-1β were lower in HG+shTUG1 group than in HG,HG+sh-Con groups(P＜0.05).Conclusions TUG1 is involved in high glucose induced pyroptosis in microglia and leads to inflammatory response.Silencing TUG1 can inhibit the pathological reaction.

Objective To investigate the effect and mechanism of Bushen Huoxue Prescription(Morindae Officinalis Radix,Cuscutae Semen,Astragali Complanati Semen,Codonopsis Radix,Poria,etc.)on endometrial hyperplasia(EH)rats of kidney deficiency and blood stasis type.Methods A total of 90 SD rats were randomly divided into normal group,model group,Mifepristone group(12 mg·kg-1),and low-,medium-,and high-dose Bushen Huoxue Prescription groups(1.2,2.4,4.8 g·kg-1),with 15 rats in each group.The EH rat model of kidney deficiency and blood stasis type was replicated by intragastric administration of Hydroxyurea and Estradiol Valerate combined with tail vein injection of high molecular dextran.After successful modeling,each group was given intragastric administration according to the set dose,once a day for 21 consecutive days.At the end of the experiment,the levels of serum cyclic adenosine monophosphate(cAMP),cyclic guanosine monophosphate(cGMP),cortisol(CORT),adrenocorticotropic hormone(ACTH),triiodothyronine(T3),thyroxine(T4),thyroid stimulating hormone(TSH),whole blood viscosity,hemorheology and coagulation function were detected by biochemical method.The levels of serum estrogen(E2),follicular estrogen(FSH),luteinizing hormone(LH),nitric oxide(NO)and endothelin-1(ET-1)were detected by ELISA.The pathological changes of endometrial tissue were observed by HE staining,and the endometrial thickness was measured.The protein expression levels of matrix metalloproteinase-9(MMP-9)and tissue inhibitor of metalloproteinase-1(TIMP-1)in endometrial tissues were detected by immunohistochemistry.The protein expression levels of MMP-9,TIMP-1,B-cell lymphoma 2(Bcl-2),Bcl-2 related X protein(Bax)and phosphatase and tensin homologue deleted on chromosome 10(PTEN)in endometrial tissue were detected by Western Blot.Results Compared with the normal group,the levels of serum CORT,ACTH,cAMP,T3,T4,TSH,ET-1 and ET-1/NO ratio in the model group were significantly decreased(P＜0.05,P＜0.01),while the levels of cGMP,E2,FSH,LH and NO were significantly increased(P＜0.05,P＜0.01).The indexes of hemorheology and fibrinogen(FIB)content were significantly increased(P＜0.05),and thrombin time(TT),prothrombin time(PT)and activated partial thromboplastin time(APTT)were significantly prolonged(P＜0.05).The uterine index and endometrial thickness were significantly increased(P＜0.01).The protein expressions of MMP-9 and Bcl-2 in endometrial tissue were significantly up-regulated(P＜0.01),and the protein expressions of TIMP-1,PTEN and Bax were significantly down-regulated(P＜0.01).Compared with the model group,the levels of serum CORT,ACTH,cAMP,T3,T4,TSH,ET-1 and ET-1/NO ratio in the Bushen Huoxue Prescription group were significantly increased(P＜0.05,P＜0.01).The levels of cGMP,E2,FSH,LH and NO were significantly decreased(P＜0.05,P＜0.01).The indexes of hemorheology and FIB content were significantly decreased(P＜0.05,P＜0.01),and TT,PT and APTT were significantly shortened(P＜0.05).The uterine index and endometrial thickness were significantly decreased(P＜0.05,P＜0.01).The protein expressions of MMP-9 and Bcl-2 in endometrial tissue were significantly down-regulated(P＜0.05,P＜0.01),and the protein expressions of TIMP-1,PTEN and Bax were significantly up-regulated(P＜0.05,P＜0.01).Conclusion Bushen Huoxue Prescription can improve the symptoms and signs of EH rats with kidney deficiency and blood stasis,protect endometrial vascular endothelial function,and alleviate endometrial hyperplasia.Its mechanism is related to the restoration of MMP-9/TIMP-1 balance,up-regulation of Bax and PTEN expressions,and inhibition of Bcl-2 expression.

Objective:To evaluate the efficacy of cerebral-placental-uterine ratio(CPUR)in predicting late-on-set fetal growth restriction(FGR).Methods:From May 2020 to May 2021,1255 women with singleton pregnancy who underwent prenatal examinations at the University of Hong Kong Shenzhen Hospital were selected for fetal growth and Doppler measurements at 35-37 +6 weeks of gestation.Pregnant women with birth weight of newbo-rns＜the 10th percentile were the FGR group.The pulsatility index(PI)of uterine artery(UtA),umbilical artery(UA)and fetal middle cerebral artery(MCA)were analyzed separately and in combination.ROC curve was used to analyze the cerebral-placental-uterine ratio(CPUR),cerebral-placental ratio(CPR),cerebral-uterine ratio(C-UtA)for predicting late-onset FGR;and to evaluate the sensitivity,positive and negative predictive value and of CPUR in the prediction of late-onset FGR.Results:The area under the curve(AUC)of CPUR,CPR,C-UtA and mean UtA-PI for FGR grope were 0.88,0.86,0.84 and 0.72.Under certain cut-off values and 87% specificity,the specificity of CPUR,CPR,C-UtA and mean UtA-Pifor predicting FGR group was 43.2%,46.6%,39.8% and 23.9%,respectively.The positive predictive values of CPUR,CPR,C-UtA and mean UtA-PI,UA-PI for predicting FGR group were 90.5%,71.9%,83.3%,63.6%and 5.2%,respectively.Conclusions:CPUR is more effective in predicting late onset FGR than CPR,C-UtA and mean UtA-PI.It can effectively increase the detection rate of fetal growth restrictionand reduce the FGR risk.