Objective : To investigate the effects of microRNA-29a(miR-29a) in mediating the regulation of dihydroartemisinin(DHA) on the immune checkpoint molecule B7H3 in lung adenocarcinoma(LUAD). Methods: The expression level and prognostic significance of B7H3 in LUAD were analyzed by public database. Small interfering RNA(siRNA) was used to knock down B7H3 in LUAD cell lines A549 and HCC827, and cell proliferation was detected by CCK-8 method. A549 and HCC827 cells were treated with gradient concentrations of DHA(0, 5, 10, 25, 50, 100 μmol/L) for 48 h, and the half maximal inhibitory concentrate(IC50) was calculated. A549 and HCC827 cells were treated with IC50concentration of DHA for 1, 2 and 3 days, and the cell proliferation was detected by CCK-8 method. A549 and HCC827 cells were transfected with miR-29a inhibitor. After DHA treatment, the expression level of miR-29a was detected by RT-qPCR, and the expression level of B7H3 was detected by Western blot. Results : B7H3 was overexpressed in LUAD and associated with poor prognosis. After knocking down of B7H3, the proliferation ability of A549 and HCC827 cells significantly decreased(allP<0.001). DHA inhibited the proliferation of A549 and HCC827 cells in both dose-and time-dependent manners, with IC50values of 30.16 μmol/L and 7.50 μmol/L, respectively. DHA up-regulated the expression of miR-29a in A549 and HCC827 cells(P<0.001,P<0.01), and down-regulated the expression of B7H3 in both cell lines(P<0.01,P<0.001). After transfection of miR-29a inhibitor into A549 and HCC827 cells, the expression of B7H3 was up-regulated, and the down-regulation of B7H3 by DHA was partially reversed. Conclusion miR-29a mediates the molecular regulation of DHA on B7H3 in LUAD.

Objective To investigate the effect of 9S management combined with plan-do-check-adjust(PDCA)cycle on the incidence of nonstandard surgical instrument handover in urological endoscopic surgery in operating room and supply room.Methods A total of 760 instruments of urological endoscopic surgery used from June 2022 to December 2022 were selected as control group and received routine management.A total of 920 instruments of urological endoscopic surgery used from January 2023 to June 2023 were selected as observation group,and 9S management and PDCA cycle management mode were administrated in instrument handover.The management performance and the satisfaction of surgeons to the instrument nursing staff were compared between the two groups.Results The accuracy of instrument identification and master rate of assembly and disassembly of instruments in the observation group were significantly higher than those in the control group(P<0.05).The incidence of instrument cleaning and packaging errors in the observation group was significantly lower than that in the control group(P<0.05).The satisfaction of surgeons to the instrument nursing staff in the observation group was higher than that in the control group,but no significant difference was found(P>0.05).Conclusion The 9S management combined with PDCA cycle management solves the problem of transport and handover of urological instruments between operating room and supply room,decreases the error rates of disassembly,cleaning,assembly and packaging of instruments,improves work efficiency,reduces the probability of instrument damage,and saves costs.

Objective To explore the influencing factors of hypothermia after extraperitoneal laparoscopic radical prostatectomy in prostate cancer patients over 80 years old,so as to improve the effectiveness of treatment.Methods The clinical data of 26 cases of prostate cancer patients over 80 years old who underwent extraperitoneal laparoscopic radical prostatectomy in Shanghai 411 hospital from January 2021 to December 2023 were analyzed retrospectively.The incidence of postoperative hypothermia was investigated.Univariate and multivariate Logistic regression analyses were used to analyze the related factors of postoperative hypothermia in elderly patients.Results The incidence of hypothermia was 61.54%(16/26).Univariate analysis indicated that body mass index(BMI),intraoperative thermal insulation,intraoperative infusion volume,operation time,and anesthesia time were related to the occurrence of postoperative hypothermia in elderly patients(all P<0.05).Multivariate Logistic regression analysis indicated that BMI≤24,intraoperative infusion volume>2 000 ml,anesthesia time>3 h and operation time>2.5 h were high risk factors for postoperative hypothermia in patients over 80 years old.Conclusion The independent influencing factors of hypothermia after extraperitoneal laparoscopic prostatectomy for selected prostate cancer patients over 80 years old are BMI,intraoperative infusion volume,duration of anesthesia,and operation time.These factors should be paid more attention during perioperative period in order to improve clinical safety.

Objective To investigate the clinical characteristics in an elderly patient with sellar multiple myeloma.Methods Clinical features,laboratory data and radiologic profile of an elderly patient with sellar multiple myeloma were collected.Results The patient was an 85-year-old male.The main clinical manifestations were fatigue,poor appetite and polyuria.Laboratory examinations showed a significant decrease in blood sodium,several anterior pitu-itary hormones and an increase in total protein,mass of pituitary lesion and concentration of prolactin.During etio-logical screening,it was found that the blood immunoglobulin G(IgG)level was significantly increased,the blood M protein was positive and the bone marrow smear showed myeloma cells accompanied by multiple osteolytic lesions in the bones of the whole body.Considering the diagnosis of multiple myeloma,the pituitary lesion was likely to be the extra-medullary involvement.Conclusions The intrasellar plasmacytoma is not common.The disease onset is insidious with clinical features and imaging findings lacking specificity.Therefore,diagnosis relies on biopsy which poses risks for elderly patients and increases diagnostic challenges leading to misdiagnosis.

Objective To evaluate the application of flipped classroom(FC)approach in endocrine system integrated course for students from 4+4 clinical medicine pilot class at Peking Union Medical College(PUMC).Methods The study included the students of 4+4 clinical medicine pilot class grades 2019-2023 in PUMC.The students of grades 2019-2021(n=77)served as the control receiving traditional teaching method,while the students of grades 2022 and 2023(n=76)were selected as the experimental group,which were taught by FC approach.The selected teaching content is thyroid theme.The scoring rates of thyroid related questions in the final exam were as-sessed and a questionnaire survey was conducted to evaluate teaching satisfaction and effectiveness.Results The scoring rates of experimental group were significantly higher as compared to that of control group(P＜0.05).Over 90％of the students in the experimental group strongly satisfied or satisfied with the teaching content arrangement,design form,classroom atmosphere,teacher-student interaction of FC and expressed willingness to continue with this methodology.In addition,over 90％of the students strongly agreed or agreed that FC stimulated learning inter-est,improved self-learning ability,strengthened the connection between theory and clinical practice,inspired clini-cal reasoning,enhanced the abilities to analyze and solve problems,and cultivated communication and teamwork skills.Conclusions The application of FC approach in endocrine system integrated course achieved excellent teaching outcomes with high satisfaction of the students.

Objective : To establish an enzyme-linked immunosorbent assay (ELISA) for exosome B7-H3 in plas- ma , and to explore the clinical significance and diagnostic value of exosome B7-H3 in plasma from non-small cell lung cancer (NSCLC) . Methods : The plasma of 70 NSCLC patients (NSCLC group) and 36 healthy controls (HC group) were collected . Exosomes and microvesicles in plasma were separated by ultra-fast centrifuge method , and the expression levels of B7-H3 in plasma exosomes in NSCLC groups and HC groups were compared by Western blot method . In NSCLC group , the expression levels of B7-H3 in plasma exosomes and microvesicles in NSCLC group were compared . A simple and feasible ELISA method was established to detect the expression level of exosome B7 - H3 in plasma by means of polyethylene glycol (PEG) precipitation and its clinical significance was analyzed . Lo- gistic regression model was established to predict plasma-derived exosome B7-H3 as a risk factor , and receiver op- erating characteristic curve (ROC) was used to investigate the diagnostic value of exosome B7-H3 in NSCLC . Results: For exosomes and microvesicles in plasma which were extracted by ultracentrifugation , Western blot results showed that the expression level of B7-H3 in plasma exosomes of NSCLC group was higher than that of HC group (P = 0. 032) , and the expression level of B7-H3 in plasma exosomes was higher than that of microvesicles of NSCLC group (P = 0. 012) . The expression level of exosome B7-H3 in plasma extracted by PEG precipitation was also higher in NSCLC group than that in HC group (P = 0. 024) . The expression level of exosome B7-H3 in plasma of NSCLC patients was not related to gender , age , smoking or pathological type , but was related to T stage (P = 0. 002) , N stage (P < 0. 001) , M stage (P = 0. 010) and AJCC stage (P < 0. 001) . Multivariate Logistic regres- sion analysis identified exosome B7-H3 in plasma as a risk factor for NSCLC . ROC analysis showed that the sensi- tivity of exosome B7-H3 in plasma for the diagnosis of NSCLC (0. 843) was higher than that of carcinoembryonic antigen (CEA) (0. 743) , whereas the specificity (0. 722) was lower than that of CEA (0. 833) . Combined de- tection of exosome B7-H3 and CEA (AUC = 0. 928 , 95% CI:0. 877 - 0. 979) had a higher diagnostic performance for NSCLC . Conclusion B7-H3 in plasma exosomes is related to the cancer staging of NSCLC , and the combined detection of exosome B7-H3 and CEA in plasma is conducive to the laboratory diagnosis of NSCLC .

Objective: To investigate the role of murine double minute 2(MDM2) in dihydroartemisinin′s(DHA) inhibition of lung adenocarcinoma cell proliferation and migration. Methods: CCK8 assay was used to detect the inhibitory effect of gradient concentrations of DHA(0, 5, 10, 25, 50 and 100 μmol/L) and time gradients(0, 24, 48, and 72 h) on the proliferation of lung adenocarcinoma A549 and PC9 cells, and the half maximal inhibitory concentrate(IC50) were calculated respectively. Colony formation and scratch assays were used to detect the inhibitory effects of DHA on colony formation and migration of A549 and PC9 cells. Western blot was used to detect the inhibitory effects of DHA on MDM2 expression and epithelial-mesenchymal transition(EMT)-related proteins E-cadherin and N-cadherin. The promoting effects of MDM2 on proliferation, migration and EMT of lung adenocarcinoma cells were verified by small interfering RNA-mediated knockdown of MDM2(si-MDM2). The reversal effects of MDM2 overexpression on DHA′s inhibition on the proliferation and migration of A549 and PC9 cells were observed. Results: DHA inhibited the proliferation of A549 and PC9 cells in a dose⁃ and time⁃dependent manner,with IC50 values of 30. 57 and 78. 61 μmol/L , respectively. Compared with the Control group , A549 and PC9 cells had significantly decreased colony formation (both P < 0. 01) and migration (both P < 0. 01) upon treatment with DHA. Moreover, DHA significantly inhibited the protein expression levels of MDM2 and N ⁃cadherin in A549 and PC9 cells , and upregulated the expression of E ⁃cadherin protein (both P < 0. 05) . Compared with si⁃Control ,si⁃MDM2 significantly inhibited the protein levels of MDM2 and N ⁃cadherin in A549 and PC9 cells , and upregulat⁃(both P < 0. 01) of both cells. After overexpression of MDM2 in A549 and PC9 cells , the proliferation and migra⁃ tion ability were significantly enhanced (both P < 0. 05) , and the inhibitory effects of DHA were partially reversed by MDM2 overexpression (both P < 0. 05) . Conclusion DHA effectively inhibits the proliferation and migration of lung adenocarcinoma cells , and its mechanism is associated with the suppression of MDM2.

ObjectiveBased on ultra performance liquid chromatography-mass spectrometry（UPLC-MS） and non-targeted metabolomics technology to discuss the central regulatory effect of Chaishao Liujuntang on chronic atrophic gastritis（CAG） rats with liver-depression and spleen-deficiency， and to look for the correlation between cerebral cortex， hypothalamus and metabolic status of gastric tissues. MethodA CAG rat model with liver-depression and spleen-deficiency was established by chemical induction， hunger and satiety disorders， chronic restraint and tail clamping stimulation， lasting for 16 weeks. Twenty-eight Wistar rats were randomly divided into a blank group of 8 rats and a model group of 20 rats. After the completion of modeling， 4 rats in the model group were taken to observe the pathological changes of gastric mucosa. The remaining model rats were randomly divided into a model group of 8 rats and a Chaishao Liujuntang group of 8 rats. Chaishao Liujuntang group rats were given 5.1 g·kg^-1 by gavage， and the remaining rats were given equal volume sterilized water by gavage for 4 weeks. Macroscopic characteristics， behavioral indicators and histopathological changes of the gastric mucosa of rats in each group were observed and compared. UPLC-MS non-targeted metabolomics was used to explore the metabolic regulation effect of Chaishao Liujuntang on the cerebral cortex， hypothalamus and stomach tissues of CAG rats with liver-depression and spleen-deficiency. Pearson correlation coefficient method was used to analyze the correlation between different tissue metabolites. ResultCompared with the model group， the macroscopic characteristics of rats in Chaishao Liujuntang group were improved， such as hair color， mental state and stool properties， and the number of times of crossing and standing in the open field experiment was significantly increased， and the static time of forced swimming was significantly reduced（P<0.01）， and the gastric mucosa atrophy was reduced. The metabolic data from the cerebral cortex of rats in each group identified a total of 3 common potential biomarkers， but not enriched in pathways， 26 common potential biomarkers were identified in the hypothalamus， and the key metabolic pathways involved were mainly enriched in purine metabolism， glycerol phospholipid metabolism， D-glutamine and D-glutamic acid metabolism. Seventeen common potential biomarkers were identified in the stomach， and the key metabolic pathways involved were mainly enriched in thiamine metabolism， valine， leucine and isoleucine biosynthesis， and taurine and taurine metabolism. Correlation analysis of metabolites in different tissues revealed that multiple amino acids and their derivatives mediated metabolic connections between the cerebral cortex， hypothalamus and stomach of rats. ConclusionThe metabolic disorders in the cerebral cortex， hypothalamus and stomach of CAG rats with liver-depression and spleen-deficiency have their own characteristics， mainly manifested by changes in the content of glycerol phospholipids， fatty acids and bile acid metabolites. Moreover， Chaishao Liujuntang may play a central regulatory role in CAG rats with liver-depression and spleen-deficiency by correcting the metabolic disorders of amino acids.

Objective To explore the effect of cannabinoid receptor 2(CB2)on orthodontic tooth movement(OTM)rate and periodontal tissue reconstruction of pressure area in mice.Methods Thirty CB2-/-male mice and thirty littermate control WT male mice were individually accepted the orthodontic appliance at their age of 6 weeks.The mice were respectively scarified at 3 days,7 days,14 days and 21 days after the operation.Then the tooth movement distance was examined through the stereomicroscope.Hematoxylin-eosin staining was performed to explore the biological responses of periodontium at the distal mesial root pressure area.Anti-tartrate acid phospha-tase staining was performed to calculate the number and distribution of osteoclasts at the distal mesial root pressure area,and MMP-9 was evaluated by immunohistochemistry to examine the number of MMP-9(+)monocytes and multinucleated cells in the same district as the TRAP staining.Results Compared with those WT mice at 3,7,14 and 21 days,OTM distance showed a gradual increased tendency according with experimental time over 21 days.The widths of periodontal ligament on the pressure side were markedly greater in CB2-/-mice than WT mice at 7,14 and 21 days(P＜0.000 1).The numbers of TRAP positive osteoclasts were significantly greater in CB2-/-mice than those in WT mice at 14 days of OTM(P＜0.001).MMP-9 immunohistochemical staining showed that the number of MMP-9(+)monocytes and multinucleated cells was more in CB2-/-mice than that in WT mice at 14 days of OTM(P＜0.05).Conclusion The absence of CB2 accelerates orthodontic tooth movement under or-thodontic force.The absence of CB2 reinforces bone resorption in orthodontic tooth movement compressive area dur-ing orthodontic tooth movement.