AIM To optimize the extraction process for Bletillae Rhizoma,and to evaluate its anti-oxidant,tyrosinase inhibitory activities.METHODS With ultrasound time,ethanol concentration and solid-liquid ratio as influencing factors,the total extraction content of gastrodin,protocatechualdehyde,p-hydroxybenzaldehyde,1,4-bis[4-(gluconoxy)benzyl]-2-isobutylmalate-2-glucoside,1,4-bis[4-(gluconoxy)benzyl]-2-isobutylmalate,yam Ⅲ,dihydropinosin and 3'-O-methylyam Ⅲ as an evaluation index,the extraction process was optimized by Box-Behnken response surface method.Subsequently,the extract's scavenging effects on DPPH,ABTS+free radicals,and inhibitory ability on tyrosinase were determined.RESULTS The optimal conditions were determined to be 49 min for ultrasound time,55％for ethanol concentration,1∶30 for solid-liquid ratio,and 2 times for extraction frequency,the total extraction content was 13.18 mg/g.The extract demonstrated the IC50 of 10.12,314.07 and 1.70 μg/g on DPPH,ABTS+free radicals and tyrosinase,respectively.CONCLUSION This simple,reliable and stable method can be used for the extraction for Bletillae Rhizoma with strong anti-oxidant,tyrosinase inhibitory activities.

Objective To analyze the outcome of modified laparoscopic uterosacral fixation in the treatment of anterior and middle pelvic organ prolapse.Methods The clinical data of 30 patients with anterior and middle pelvic organ prolapse undergoing modified laparoscopic uterosacral fixation in our hospital from September 2020 to July 2023 were analyzed retrospectively.The perioperative related indicators and complications of patients during follow-up were observed.The pelvic organ prolapse quantification(POP-Q)scores,pelvic floor distress inventory-short form 20(PFDI-20)scores,pelvic floor impact questionnaire-short form 7(PFIQ-7)scores,and patient global impression of improvement(PGI-I)scores of patients before and after operation were compared.Results The operations of all patients were successfully completed without conversion to open surgery or serious complications.The average operation time was(170.8±21.5)minutes,the average intraoperative blood loss was(29.3±20.2)mL,the average postoperative exhaust time was(1.7±0.7)days,the average postoperative retaining urinal catheter time was(2.3±0.6)days,and the average postoperative hospitalization time was(4.2±0.8)days.Except for the total vaginal length,other indicators of POP-Q of patients 3 and 6 months after operation were significantly improved compared with those before operation(P<0.05).The PFDI-20 score and PFIQ-7 score of patients 6 months after operation were significantly improved compared with those before operation(P<0.05).The objective cure rate of patients was 100%;the results of PGI-I questionnaire showed that the subjective success rate of patients was 100%.Conclusion Modified laparoscopic uterosacral fixation is an effective operation for the treatment of anterior and middle pelvic organ prolapse,which has the advantages of good anatomical recovery,high satisfaction,and few postoperative complications,with a definite short-term curative effect.

Objective To analyze the outcome of modified laparoscopic uterosacral fixation in the treatment of anterior and middle pelvic organ prolapse.Methods The clinical data of 30 patients with anterior and middle pelvic organ prolapse undergoing modified laparoscopic uterosacral fixation in our hospital from September 2020 to July 2023 were analyzed retrospectively.The perioperative related indicators and complications of patients during follow-up were observed.The pelvic organ prolapse quantification(POP-Q)scores,pelvic floor distress inventory-short form 20(PFDI-20)scores,pelvic floor impact questionnaire-short form 7(PFIQ-7)scores,and patient global impression of improvement(PGI-I)scores of patients before and after operation were compared.Results The operations of all patients were successfully completed without conversion to open surgery or serious complications.The average operation time was(170.8±21.5)minutes,the average intraoperative blood loss was(29.3±20.2)mL,the average postoperative exhaust time was(1.7±0.7)days,the average postoperative retaining urinal catheter time was(2.3±0.6)days,and the average postoperative hospitalization time was(4.2±0.8)days.Except for the total vaginal length,other indicators of POP-Q of patients 3 and 6 months after operation were significantly improved compared with those before operation(P<0.05).The PFDI-20 score and PFIQ-7 score of patients 6 months after operation were significantly improved compared with those before operation(P<0.05).The objective cure rate of patients was 100%;the results of PGI-I questionnaire showed that the subjective success rate of patients was 100%.Conclusion Modified laparoscopic uterosacral fixation is an effective operation for the treatment of anterior and middle pelvic organ prolapse,which has the advantages of good anatomical recovery,high satisfaction,and few postoperative complications,with a definite short-term curative effect.

AIM To investigate the ameliorative effects of Compound Fufangteng Mixture(CFM)on cyclophosphamide(CTX)-induced immunosuppression in mice.METHODS Forty-eight male C57BL/6J mice were randomly divided into the blank control group,the model group,the levamisole hydrochloride group(40 mg/kg)and the low-dose,medium-dose and high-dose CFM groups(3.75,7.5,10 g/kg),with 8 mice in each group,and given respective intervention orally once daily for 14 days.On the 5th to 7th day of administration,with the blank control group given normal saline intraperitoneally,the other groups underwent intraperitoneal CTX injections(80 mg/kg).24 hours after the last administration,organ indices of thymus and spleen were calculated;splenic histopathological alterations were assessed by HE staining;serum levels of IL-2,IL-6 and IgG were quantified using ELISA;splenic CD4+,CD8+T lymphocytes,alongside CD86+and CD206+macrophages populations were analyzed by flow cytometry;and splenic expression of CD4,CD8 and F4/80 was evaluated by immunohistochemical staining.RESULTS In CTX-treated mice,CFM administration mitigated body weight loss;enhanced thymus weight and thymic index;ameliorated splenic immune cell populations,elevated serum levels of cytokines IL-2,IL-6 and IgG in serum;and upregulated splenic levels of CD45+CD3+T lymphocytes and F4/80+CD11b+macrophages,alongside increasing the expression of CD4,CD8 and F4/80 surface markers.CONCLUSION CFM alleviates CTX-induced immunosuppression state in mice by modulating immune cells,restoring immune function and enhancing anti-inflammatory and tissue repair capabilities.

Sepsis is a life-threatening organ dysfunction caused by a dysregulated host response to infection,with a high mortality rate.Sirtuin 3,a deacetylase within mitochondria,plays an important regulatory role in cellular metabolism,oxidative stress,and inflammatory responses.In recent years,significant progress has been made in the study of the function and regulatory role of sirtuin 3 in sepsis-related diseases.Research has shown that sirtuin 3 can alleviate organ damage caused by sepsis by regulating mitochondrial function,reducing oxidative stress,and inhibiting inflammatory responses.The specific mechanisms include the regulation of mitochondrial bioenergetics,activation of antioxidant enzyme systems,and inhibition of inflammatory mediator expression.In addition,sirtuin 3 plays a protective role in the pathological process of sepsis by interacting with multiple signaling pathways.This article summarizes the functions and regulatory mechanisms of sirtuin 3 in various sepsis-related diseases,aiming to provide new targets and strategies for the prevention and treatment of sepsis in the future.
Sepsis/metabolism* ; Sirtuin 3/physiology* ; Humans ; Animals ; Oxidative Stress ; Mitochondria/metabolism* ; Signal Transduction

AIM To investigate the ameliorative effects of Compound Fufangteng Mixture(CFM)on cyclophosphamide(CTX)-induced immunosuppression in mice.METHODS Forty-eight male C57BL/6J mice were randomly divided into the blank control group,the model group,the levamisole hydrochloride group(40 mg/kg)and the low-dose,medium-dose and high-dose CFM groups(3.75,7.5,10 g/kg),with 8 mice in each group,and given respective intervention orally once daily for 14 days.On the 5th to 7th day of administration,with the blank control group given normal saline intraperitoneally,the other groups underwent intraperitoneal CTX injections(80 mg/kg).24 hours after the last administration,organ indices of thymus and spleen were calculated;splenic histopathological alterations were assessed by HE staining;serum levels of IL-2,IL-6 and IgG were quantified using ELISA;splenic CD4+,CD8+T lymphocytes,alongside CD86+and CD206+macrophages populations were analyzed by flow cytometry;and splenic expression of CD4,CD8 and F4/80 was evaluated by immunohistochemical staining.RESULTS In CTX-treated mice,CFM administration mitigated body weight loss;enhanced thymus weight and thymic index;ameliorated splenic immune cell populations,elevated serum levels of cytokines IL-2,IL-6 and IgG in serum;and upregulated splenic levels of CD45+CD3+T lymphocytes and F4/80+CD11b+macrophages,alongside increasing the expression of CD4,CD8 and F4/80 surface markers.CONCLUSION CFM alleviates CTX-induced immunosuppression state in mice by modulating immune cells,restoring immune function and enhancing anti-inflammatory and tissue repair capabilities.

AIM To optimize the extraction process for Bletillae Rhizoma,and to evaluate its anti-oxidant,tyrosinase inhibitory activities.METHODS With ultrasound time,ethanol concentration and solid-liquid ratio as influencing factors,the total extraction content of gastrodin,protocatechualdehyde,p-hydroxybenzaldehyde,1,4-bis[4-(gluconoxy)benzyl]-2-isobutylmalate-2-glucoside,1,4-bis[4-(gluconoxy)benzyl]-2-isobutylmalate,yam Ⅲ,dihydropinosin and 3'-O-methylyam Ⅲ as an evaluation index,the extraction process was optimized by Box-Behnken response surface method.Subsequently,the extract's scavenging effects on DPPH,ABTS+free radicals,and inhibitory ability on tyrosinase were determined.RESULTS The optimal conditions were determined to be 49 min for ultrasound time,55％for ethanol concentration,1∶30 for solid-liquid ratio,and 2 times for extraction frequency,the total extraction content was 13.18 mg/g.The extract demonstrated the IC50 of 10.12,314.07 and 1.70 μg/g on DPPH,ABTS+free radicals and tyrosinase,respectively.CONCLUSION This simple,reliable and stable method can be used for the extraction for Bletillae Rhizoma with strong anti-oxidant,tyrosinase inhibitory activities.

Objective:To explore the function of MDM2 and its relationship with p53 at the cellular level during H 2O 2 induced oxidative damage. Methods:MLE-12 HALI cell models were established using 0.5 mmol/L H 2O 2, and were divided into three groups: normal control group, H 2O 2 injury group, H 2O 2+MDM2 overexpressed group, and H 2O 2+MDM2 shRNA group. Infection of MLE-12 cells with adenovirus vector overexpressing and silencing MDM2; Using immunoprecipitation (Co-IP) to analyze the interaction between MDM2 and p53; Western blotting was used to detect the protein expression levels of MDM2, p53, Bcl-2, Bax, and cleared caspase-3 after HALI modeling; Measure the apoptosis rate of cells in each group. Results:After transcriptome sequencing,the p53 signaling pathway closely related to HALI. Compared with the normal group, the expression of MDM2 in the H 2O 2 injury group was lower ( P<0.05); Compared with the H 2O 2 injury group, overexpression of MDM2 resulted in a decrease in the apoptosis rate of MLE-12 cells ( P<0.05), a decrease in the expression levels of p53, Bax, and cleared caspase-3 proteins, and an upregulation of MDM2 and Bcl-2 protein expression ( P<0.05). Compared with the H 2O 2 injury group, when MDM2 was silenced, the cell apoptosis rate increased ( P<0.05), and the expression levels of p53, Bax, and cleared caspase-3 proteins were upregulated, while the expression levels of MDM2 and Bcl-2 proteins decreased ( P<0.05). Co-IP experiments showed that MDM2 binds to p53 protein. Conclusions:MDM2 can exert a protective effect on HALI by inhibiting MLE-12 cell apoptosis through the p53/Bcl-2/Bax axis.

Objective To investigate the impact of Maxing Shigan decoction on lung tissue damage in cough variant asthma(CVA)rats by regulating the Notch1/Jagged1 signaling pathway.Methods The CVA rat model was established by ovalbumin+aluminum hydroxide sensitization method.After successful modeling,the rats were randomly divided into control group(conventional feeding),model group(4％ovalbumin 0.5 mL+2％aluminum hydroxide 0.2 mL),combined group(on the basis of the model group,16 g·kg-1 Maxing Shigan decoction and 200 mg·kg-1 Jagged1 were added respectively)and experimental-L,experimental-M,experimental-H groups(on the basis of the model group,4,8 and 16 g·kg-1 Maxing Shigan decoction were added respectively),with 15 rats in each group.The cough frequency and histopathological changes of the lung tissue were observed,and the expression level of Notch1 and Jagged1 protein were detected by Western blot.Results The coughing times of rats in control group,model group,experiment-L group,experiment-M group,experiment-H group and combined group were(2.04±0.08),(9.21±0.82),(7.38±0.60),(5.44±0.42),(3.14±0.32)and(7.88±0.65)times,respectively;the number of goblet cells in lung tissue was 13.65±1.24,59.66±5.54,47.21±4.12,34.58±3.01,23.54±2.06 and 39.25±3.68,respectively;the relative expression levels of Notch1 protein were 0.42±0.03,1.16±0.09,0.97±0.09,0.76±0.07,0.59±0.05 and 0.89±0.07,respectively;the relative expression levels of of Jagged1 protein were 0.36±0.03,1.09±0.04,0.92±0.09,0.70±0.07,0.46±0.04 and 0.79±0.07,respectively.The above indexes of the model group were compared with those of the control group,the experimental-L group,the experimental-M group and the experimental-H group,and the above indexes of the combined group were compared with those of the experimental-H group,and the differences were statistically significant(all P＜0.05).Conclusion Maxing Shigan decoction may improve lung tissue damage in CVA rats by inhibiting the activation of Notch1/Jagged1 signaling pathway.

Objective To investigate the impact of exosomal(Exos)-miR-21-5p(miR-21)targeting S-phase kinase associated protein 2(SKP2)derived from Type Ⅱ alveolar epithelial cells(AEC-Ⅱ)on the patho-genesis of bronchopulmonary dysplasia(BPD).Methods A total of 60 SD rats aged 6～8 weeks were utilized in this study,with 30 of them subjected to extraction and culture through differential adherent centrifugation.Density gradient centrifugation was employed for the isolation of AEC-Ⅱ derived exosomes,while vesicles from AEC-Ⅱ me-dium were extracted using density gradient centrifugation.These isolates were subsequently confirmed by transmission electron microscopy and particle size analysis,and the targeting relationship between miR-21 and SKP2 was validated through dual-fluorescein reporter gene assay.The remaining 30 mice were combined in a male-to-female ratio of 3:1 to facilitate pregnancy testing.These neonatal mice were randomly assigned into four experimental groups:air control group(Con group),hyperoxia group(BPD+PBS group),hyperoxia-treated mice receiving exosomes(BPD+Exos-miR-21 group),and hyperoxia combined with exosome miR-21 inhibitor treatment group(BPD+Exos-AV-miR-21 group).Neonatal SD rats will be exposed to 85％oxygen to establish a BPD model.Follow-ing 14 days of high oxygen treatment,the expression levels of miR-21 in lung tissues and exosomes will be assessed using RT-qPCR.HE staining will be employed to observe pathological changes in lung tissue,while mean alveolar linear intercept(MLI)and radial alveolar count(RAC)will be calculated.Superoxide dismutase(SOD),malondialdehyde(MDA),and total antioxidant capacity(T-AOC)levels will be determined spectrophoto-metrically,whereas reactive oxygen species(ROS)levels will be measured via fluorescence spectrophotometry.Additionally,Western blot analysis will assess the expression levels of SKP2,NR2F2,and VEGF-A proteins.Results The results obtained from electron microscopy and particle size analysis demonstrated that the vesicle structure isolated from AEC-Ⅱ cells corresponded to exosomes.Moreover,there was a significant upregulation of miR-21 expression in exosomes(P＜0.01).Subsequently,the dual luciferase gene reporter assay con-firmed SKP2 as the target of miR-21.Comparative analysis revealed that compared to the Con group,both BPD+PBS and BPD+Exos-AV-miR-21 groups exhibited disordered lung tissue structure with enlarged and simpli-fied alveoli,increased levels of ROS,MDA,and MLI along with elevated expression of SKP2 protein(P＜0.01).Conversely,RAC,SOD,T-AOC levels were downregulated alongside miR-21 expression while NR2F2 and VEGF-A protein expressions decreased significantly(P＜0.01).In contrast to the BPD+PBS group,the number of alveoli without alveoli increased in the BPD+Exos-miR-21 group leading to improved degree of alveolar simplification accom-panied by reduced MLI,ROS,MDA levels as well as decreased SKP2 protein expression(P＜0.01).Addition-ally ROC,SOD,T-AOC,and miR-21 expressions were upregulated while NR2F2and VEGF-A expressions were increased(P＜0.01).Conclusions The exosomal miR-21 derived from AEC-Ⅱ may potentially target SKP2,thereby inhibiting oxidative stress and promoting alveolar development.Consequently,it can improve BPD by enhancing the protein expression of NR2F2 and VEGF-A.