Twelve compounds were isolated from the rice fermentation extracts of Penicillium expansum GY618 by silica column chromatography, Sephadex gel column chromatography, ODS column chromatography and semi preparative HPLC methods. They were determined as 11-hydroxyl-penicitrinone F (1), penicitrinone F (2), betulin (3), erythrodiol (4), ergosterol (5), ergost-5α,8α-epidioxy-6,22-dien-3β-ol (6), (5α,8α-epidioxy-(22E,24R)-ergosta-6,9(11),22-trien-3β-ol) (7), 5α,8α-epidioxy-(22E,24R)-23-methylergosta-6,22-dien-3β-ol (8), 5α,8α-epidioxy- 23,24(R)-dimethylcholesta-6,9(11),22-trien-3β-ol (9), dankasterone A (10), (17R)-4-hydroxy-17-methylincisterol (11) and ergosta-4,6,8(14),22-tetraen-3-one (12), through mass spectrometer, nuclear magnetic resonance (NMR) and comparison with the literature. Compound 1 was a new compound and compounds 2-4, 6-12 were isolated from Penicillium expansum fungus for the first time. The tyrosinase inhibitory activity experiment showed that compounds 1, 3 and 12 showed certain inhibitory activity against tyrosinase with IC₅₀ values of (75 ± 9), (69 ± 8) and (64 ± 2) μmol·L^-1, respectively. The IC₅₀ of other compounds were all greater than 100 μmol·L^-1, while IC₅₀ of the positive control kojic acid was (46 ± 4) μmol·L^-1.

Aim To screen and study the effects of Tao Hong Si Wu decoction(THSWD)-mediated treat-ment on circular RNA(circRNA)expression profiles in rats with middle cerebral artery occlusion(MCAO),and investigate the possible roles and molecular mecha-nisms of THSWD.Methods Next-generation RNA sequencing was conducted to identify circRNA expres-sion profiles in MCAO rats after treatment with THSWD and compared with the MCAO model group and control group.Bioinformatics analysis was performed to predict the potential target microRNAs and mRNAs.Gene On-tology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses for the potential target mRNAs were applied to explore the potential roles of differentially expressed circRNAs.RT-qPCR was performed to verify circRNAs with significant differences in expression.Results We identified 87 significantly differentially expressed circRNAs between the MCAO group versus the control group,and 86 sig-nificantly differentially expressed circRNAs between the MCAO group versus the THSWD group.respective-ly.Among them,17 circRNAs induced by the MCAO model were reversed via treatment with THSWD.To demonstrate the roles of mRNAs targeted by DECs,the GO and KEGG databases were used.Further analysis revealed that five circRNAs may play important roles in the development of MCAO.Conclusions The com-prehensive expression profile of circRNAs in rats with middle cerebral artery occlusion after THSWD treat-ment is determined for the first time,suggesting that the therapeutic effect of THSWD on MCAO may be a-chieved by regulating the expression of circRNAs.

Aim To explore the improvement effect of Bazi Bushen capsule on thymic degeneration in SAMP6 mice and the possible mechanism.Methods Twenty 12 week old male SAMP6 mice were randomly divided into the model group(SAMP6)and the Bazi Busheng capsule treatment group(SAMP6+BZBS).Ten SAMR1 mice were assigned to a homologous control group(SAMR1).The SAMP6+BZBS group was oral-ly administered Bazi Bushen capsule suspension(2.8 g·kg-1)daily,while the other two groups were orally administered an equal amount of distilled water.After nine weeks of administration,the morphology of the thymus in each group was observed and the thymus in-dex was calculated;HE staining was used to observe the structural changes of thymus tissue;SA-β-gal stai-ning was used to detect thymic aging;flow cytometry was used to detect the proportion of thymic CD3+T cells in each group;Western blot was used to detect the levels of p16,Bax,Bcl-2,and cleaved caspase-3 proteins in thymus;immunofluorescence was applied to detect the proportion of cortical thymic epithelial cells in each group;ELISA was employed to detect IL-7 lev-els in thymus.Results Compared with the SAMP6 group,the thymic index of the SAMP6+BZBS group significantly increased(P＜0.05);the disordered thy-mic structure was significantly improved;the positive proportion of SA-β-gal staining significantly decreased(P＜0.01);the proportion of CD3+T cells apparently increased(P＜0.05);the level of p16 protein signifi-cantly decreased(P＜0.05);the level of Bcl-2 pro-tein significantly increased(P＜0.05),while the lev-el of cleaved caspase-3 protein markedly decreased(P＜0.05);the proportion of cortical thymic epithelial cells evidently increased;the level of IL-7 significantly increased(P＜0.01).Conclusions Bazi Bushen capsule can delay thymic degeneration,inhibit cell ap-optosis in thymus and promote thymic cell development in SAMP6 mice,which may be related to increasing the proportion of cortical thymic epithelial cells and promoting IL-7 secretion.

Objective To compare the efficacy of renal proximal renal artery denervation(pRDN)and full-length renal artery denervation(fRDN)for treatment of hypertension.Methods Fifty-six hypertensive patients were enrolled and randomly assigned to full-length renal artery denervation group(n=25)and proximal renal artery denervation group(n=31).After the procedure,24-hour ambulatory blood pressure monitoring(24 h-ABPM)at 6 months and office blood pressure at 12 months was recorded for statistical analysis.Results The blood pressure at follow-up reduced significantly in both groups,while there was no significant difference between groups.The baseline office blood pressure in fRDN group and pRDN group was(180±15)/(104±10)mmHg and(180±12)/(103±8)mmHg,respectively,which decreased to(142±9)/(82±7)mmHg and(143±10)/(83±6)mmHg at 12 months postoperatively(P＜0.001 within groups and P＞0.05 between groups).The baseline 24 h-ABPM in the two groups was(162±13)/(95±8)mmHg and(160±12)/(94±8)mmHg,respectively,which decreased to(142±11)/(83±7)mmHg and(141±8)/(81±7)mmHg at 6 months postoperatively(P＜0.001 within groups and P＞0.05 between groups).However,there was no significant difference in the reduction of office blood pressure and ambulatory blood pressure between the two groups.No treatment-related adverse events were observed.Conclusions pRDN has similar antihypertensive effect to fRDN.

The feasibility of fecal-oral transmission of Trichinella spiralis larva and infectivity in feces were assessed.Six Wistar rats were infected with 4 000 T.spiralis larvae at one time.The feces of three rats(group A)were collected at 0,4,8,12,16,24,and 48 h,and then divided into four aliquots and stored at room temperature for 0,24,48,and 72 h,respectively.The number of larvae in the feces was counted under a microscope,and each aliquot containing T.spiralis larvae was fed to five Kunming mice.All mice were killed 42 days after infection.The number and reproductive capacity index(RCI)of T.spir-alis larva were calculated.The feces from another three Wistar rats(group B)were collected from 1 to 23 days after infection and DNA was extracted from each fecal sample for PCR detection of the mitochondrial atp6 gene of T.spiralis.The results showed that T.spiralis was discharged in feces at 4-16 h after infection,peaking at 8 h,with no detection after 24 h.The numbers of T.spiralis larva in fecal samples of three rats in group A were 350,400,145,and 40 at 4,8,12,and 16 h after infection,respectively.The RCI values of T.spiralis larva for samples collected at 4 h were 112.5,20.5,and 2 after storage at room temperature for 0,24,and 48 h,66.7 and 9 for samples collected at 8 h after 0 and 24 h,13.3 and 5 for samples collected at 12 after 0 and 24 h,and 10 for samples collected at 16 h after 0 h,respectively.Of the three rats in group B,two were posi-tive for the atp6 gene for 18 consecutive days,and the third was positive for 20 consecutive days.These results indicate that one-time ingestion of T.spiralis larva in quantity can discharge infectious T.spiralis larvae,which can be transmitted through the fecal-oral route.PCR analysis is appropriate for detection of the T.spiralis atp6 gene in feces,but not infectivity of T.spiralis.

Objective:To evaluate the role of sphingosine-1-phospho-1 receptor 1 (S1PR1) in remifentanil-induced hyperalgesia in rats with incisional pain and the relationship with KCNQ2/3 potassium channels in the dorsal root ganglia (DRG).Methods:Forty-eight male Sprague-Dawley rats with successful caudal vein catheterization, aged 2-3 months, weighing 260-280 g, were divided into 6 groups ( n=8 each) using a random number table method: control group (group C), S1PR1 inhibitor group (FTY720) group (group F), remifentanil group (group R), remifentanil + S1PR1 inhibitor (FTY720) group (group RF), remifentanil + incision pain group (group RI) and remifentanil + incision pain + S1PR1 inhibitor (FTY720) group (group RIF). In group C, normal saline 0.1 ml· kg -1·min -1 was intravenously infused for 60 min. In group F, FTY720 3 nmol was intrathecally injected at 10 min before normal saline injection, and 0.1 ml · kg -1·min -1 normal saline was infused into the caudal vein for 60 min. Remifentanil 1.0 μg· kg -1·min -1 was infused for 60 min through the caudal vein in group R. In RF group, FTY720 (3 nmol) was intrathecally injected, and 10 min later remifentanil 1.0 μg· kg -1·min -1 was infused via the caudal vein for 60 min. The incisional pain model was established, and remifentanil 1.0 μg· kg -1·min -1 was infused via the caudal vein for 60 min in RI group. In RIF group, FTY720 3 nmol was intrathecally injected at 10 min before remifentanil infusion, then the incisional pain model was developed, and remifentanil 1.0 μg· kg -1·min -1 was infused via the caudal vein at the same time for 60 min. The mechanical paw withdraw threshold (MWT) and thermal paw withdraw latency (TWL) were measured at 24 h before remifentanil or normal saline infusion (T 0) and 2, 6, 24 and 48 h after remifentanil or normal saline infusion (T 1-4). The rats were sacrificed after the last measurement of pain threshold, and the L 4-6 segments of the DRG were taken for determination of the expression of S1PR1, KCNQ2 and KCNQ3 protein and mRNA in the DRG by Western blot and real-time polymerase chain reaction. Results:Compared with group C, the MWT was significantly decreased, and the TWL was shortened at T 1-4, the expression of S1PR1 protein and mRNA in the DRG was up-regulated, the expression of KCNQ2 and KCNQ3 protein and mRNA in the DRG was down-regulated ( P<0.05), and no significant change was found in each parameter in R and RI groups ( P>0.05). Compared with group R, the MWT was significantly decreased, and the TWL was shortened at T 1-4, the expression of S1PR1 protein and mRNA in the DRG was up-regulated, and the expression of KCNQ2 and KCNQ3 protein and mRNA in the DRG was down-regulated in group RI, and the MWT was significantly increased, and the TWL was prolonged at T 1-4, the expression of S1PR1 protein and mRNA in the DRG was down-regulated, and the expression of KCNQ2 and KCNQ3 protein and mRNA in the DRG was up-regulated in group RF ( P<0.05). Compared with group RI, the MWT was significantly increased, and the TWL was prolonged at T 1-4, the expression of S1PR1 protein and mRNA in the DRG was down-regulated, and the expression of KCNQ2 and KCNQ3 protein and mRNA in the DRG was up-regulated in group RIF ( P<0.05). Conclusions:S1PR1 is involved in the process of remifentanil-induced hyperalgesia in rats with incisional pain, which is related to the inhibition of KCNQ2/3 potassium channel expression in the DRG.

Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75％(27/36);boys accounted for 64％(23/36).Among the 36 neonates,31(86％)had disease onset within 3 days after birth,and 19(53％)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61％)had left cerebral infarction and 13(36％)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75％)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72％)and seizures in 10 neonates(34％).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33％,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44％(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.

Aim To screen and study the expression of long non-coding RNA (IncRNA) in rats with middle cerebral artery occlusion (MCAO) with MCAO treated with Tao Hong Si Wu decoction (THSWD) and determine the possible molecular mechanism of THSWD in treating MCAO rats. Methods Three cerebral hemisphere tissue were obtained from the control group, MCAO group and MCAO + THSWD group. RNA sequencing technology was used to identify IncRNA gene expression in the three groups. THSWD-regulated IncRNA genes were identified, and then a THSWD-regu-lated IncRNA-mRNA network was constructed. MCODE plug-in units were used to identify the modules of IncRNA-mRNA networks. Gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) were used to analyze the enriched biological functions and signaling pathways. Cis- and trans-regulatory genes for THSWD-regulated IncRNAs were identified. Reverse transcription real-time quantitative pol-ymerase chain reaction (RT-qPCR) was used to verify IncRNAs. Molecular docking was used to identify IncRNA-mRNA network targets and pathway-associated proteins. Results In MCAO rats, THSWD regulated a total of 302 IncRNAs. Bioinformatics analysis suggested that some core IncRNAs might play an important role in the treatment of MCAO rats with THSWD, and we further found that THSWD might also treat MCAO rats through multiple pathways such as IncRNA-mRNA network and network-enriched complement and coagulation cascades. The results of molecular docking showed that the active compounds gallic acid and a-mygdalin of THSWD had a certain binding ability to protein targets. Conclusions THSWD can protect the brain injury of MCAO rats through IncRNA, which may provide new insights for the treatment of ischemic stroke with THSWD.

Objective To observe the clinical efficacy of Modified Tongxie Yaofang(Important Formula for Relieving Diarrhea with Pain)in the treatment of patients with diarrhea-predominant irritable bowel syndrome(IBS-D)with liver-depression and spleen-deficiency syndrome.Methods Sixty patients with IBS-D of liver depression and spleen deficiency type were randomly divided into treatment group and control group,30 patients in each group.The patients in the treatment group were given modified Tongxie Yaofang,and the patients in the control group were given Pivacurium Bromide Tablets(Dicetel).Both groups were treated for a 4-week course of treatment.The changes of traditional Chinese medicine(TCM)syndrome scores,irritable bowel syndrome quality of life(IBS-QOL)scores,Hamilton Anxiety Scale(HAMA)scores of the two groups were observed before and after the treatment.Moreover,the efficacy for overall symptomatic improvement in the two groups was evaluated according to the grading of irritable bowel syndrome symptom severity score(IBS-SSS),and the efficacy for improving single symptom such as abdominal pain and diarrhea as well as the relapse after 4 weeks of drug cessation in the two groups were compared.Results(1)After 4 weeks of treatment,the total effective rate for overall symptomatic improvement in the treatment group was 83.33%(25/30),and that in the control group was 43.33%(13/30).The intergroup comparison(tested by chi-square test)showed that the efficacy for overall symptomatic improvement in the treatment group was significantly superior to that in the control group(P＜0.01).(2)After 4 weeks of treatment,the total effective rate for improving single symptom of abdominal pain and diarrhea in the treatment group was 80.00%(24/30),90.00%(27/30),and that in the control group was 43.33%(13/30),46.67%(14/30),respectively.The intergroup comparison(tested by chi-square test)showed that the efficacy for improving single symptom of abdominal pain and diarrhea in the treatment group was significantly superior to that in the control group(P＜0.01).(3)After 4 weeks of treatment,the TCM syndrome scores,IBS-QOL scores,and HAMA scores of patients in both groups were significantly decreased compared with those before treatment(P＜0.05 or P＜0.01),and the decrease in the treatment group was significantly superior to that in the control group(P＜0.01).(4)After 4 weeks of drug withdrawal,the recurrence rate of the treatment group was 24.00%(6/25),which was significantly lower than that of the control group(61.54%,8/13),and the difference was statistically significant between the two groups(P＜0.05).Conclusion Modified Tongxie Yaofang exerts certain effect in treating patients with IBS-D of liver depression and spleen deficiency type,and the decoction is effective on decreasing the scores of symptoms of abdominal pain and diarrhea as well as TCM syndrome scores,improving the quality of life of the patients,and alleviating the anxiety status of the patients.

Histone acetylation and methylation can affect chromatin conformation and regulate a variety of biological activities.Abnormal histone acetylation and methylation modifications are related to the occurrence and development of a variety of oral diseases.Histone acetylation and methylation increase or decrease in an orderly manner to regulate the development of teeth.Fluoride ions can destroy the balance between histone acetylation and methylation,which may be related to the occurrence of dental fluorosis.In addition,histone acetylation and methylation are involved in the regula-tion of oral inflammatory diseases.In the inflammatory microenvironment,the expression of histone acetyltransferase GCN5 decreases,and the expression of Dickkopf 1(DKK1)decreases,activating the Wnt/β-catenin pathway and ulti-mately inhibiting the osteogenic differentiation of periodontal ligament stem cells.Enhancer of zeste homolog 2(EZH2)and H3K27me3 levels were decreased in inflamed dental pulp tissues and cells.EZH2 inhibition inhibited the expres-sion of interleukin(IL)-1b,IL-6 and IL-8 in human dental pulp cells under inflammatory stimulation.Histone acetyla-tion/methylation modifications can interact with multiple signaling pathways to promote the occurrence and development of oral tumors and are related to the high invasiveness of salivary gland tumors.Small molecule drugs targeting histone acetylation and methylation-related enzymes can regulate the level of histone methylation/acetylation and have shown po-tential in the treatment of oral and maxillofacial diseases.For example,the histone deacetylase inhibitor vorinostat can inhibit the secretion of inflammation-related cytokines;it also promotes the maturation of odontoblasts and the formation of dentin-related matrix,demonstrating its potential in pulp preservation.Understanding the role of histone acetylation/methylation modifications in the occurrence and development of oral diseases will help promote research on epigenetic modifications in oral diseases and provide new perspectives for disease diagnosis and treatment.