Objective To study the immunoregulatory effects of emodin on macrophages during Brucella infection and to provide theoretical and experimental basis for developing new drugs to treat brucello-sis. Methods Bone marrow cells were isolated from BALB/c mice and cultured with MG-CSF to induce differentiation. Flow cytometry was used to detect the differentiation of bone marrow cells into macrophages (MΦ) by using FITC-labeled mouse anti-F4/80 antibody and PE-labeled anti-CD11b antibody. MTT meth-od was used to detect the influences of various concentrations of emodin on the survival rate of MΦ. Doxy-cycline was used as the control to compare half maximal inhibitory concentrations (IC50) of the two drugs. MΦ were cultured with Brucella at a ratio of 100 : 1 for 4 h. MΦ and Brucella were further cultured for 1, 6,12,24 and 48 h after adding emodin. Effects of emodin on the survival of MΦ were analyzed by colony counting method. ELISA was performed to detect the levels of TNF-α, IL-6 and IFN-γ in culture superna-tants. Results On day 8 of culturing,91.28% of bone marrow cells differentiated into macrophages. The IC50of emodin(608.4 μg/ml) was significantly higher than that of doxycycline(225.5 μg/ml). The logC-FU values of emodin stimulation groups (6,12,24 and 48 h) were significantly lower than those of blank control groups. Among all emodin stimulation groups, the 24 h group had the lowest logCFU value, which was also lower than that of the doxycycline treatment group. The levels of TNF-α,IL-6 and IFN-γ in 6,12 and 24 h emodin stimulations group increased significantly as compared with those of the corresponding con-trol groups. The levels of TNF-α, IL-6 and IFN-γ peaked at 24 h of culturing Brucella-infected MΦ with emodin. No significant difference in IFN-γ level was found between the 12 and 24 h emodin stimulation groups [(74.233 ±4.416) pg/ml vs (78.328 ±8.932) pg/ml]. Conclusion Emodin may enhance the ability of macrophages to kill Brucella through promoting the expression of TNF-α,IL-6 and IFN-γ.

Communication between doctors and patients is an important way to establish good doctorpatient relationship.It is crucial to master doctor-patient communication skills and arts for general practitioners who will extensively serve the people in community for life-long time after completion of residency training.To improve the communication skills and to enhance the clinical competency of general practitioners,we applied the Balint special group activities as core of doctor-patient communication course in residency training program.Through 3 years of practice,we found that the application of Balint group enriched the teaching contents and form of general practice residency training;improved doctor-patient communication skills,and enhanced the competency and professionalism of general practice trainees.

ObjectiveTo investigate the possible mechanism by which Zhiqiao Gancao Decoction （枳壳甘草汤， ZGD） delays intervertebral disc degeneration （IDD） based on the Hippo-yes-associated protein （YAP）/transcriptional co-activator with PDZ-binding motif （TAZ） signaling pathway. MethodsA total of 50 SD rats were randomly divided into sham surgery group， model group， low-dose ZGD group， high-dose ZGD group， and high-dose ZGD + inhibitor group， with 10 rats in each group. In the sham surgery group， the rats were pierced in the skin and muscle at the Co6/7/8 segments of the tail with a 21G needle （depth approximately 2 mm） without damaging the intervertebral disc. In the other groups， rats were injected with a 21G needle at the Co6/7/8 segments of the tail to establish an IDD model by piercing the tail intervertebral disc 5 mm. One week after modeling， rats in the low-dose and high-dose ZGD groups were given 6.24 and 12.24 g/（kg·d） of the decoction via gastric gavage， respectively. The high-dose ZGD + inhibitor group was given 12.24 g/（kg·d） of the decoction and an intraperitoneal injection of YAP/TAZ inhibitor Verteporfin 10 mg/kg. The sham surgery and model groups were given 5 ml/（kg·d） of normal saline via gavage. The gavage was given once a day， and the intraperitoneal injection was given every other day. After 4 weeks of continuous intervention， the pathological changes of the tail intervertebral discs were observed using HE staining， Oil Red O-Green staining， and Toluidine Blue staining. Immunohistochemistry was used to detect the expression of aggrecan and MMP3 in the nucleus pulposus. TUNEL fluorescence staining was performed to detect apoptosis in the nucleus pulposus， and the apoptosis rate was calculated. Western blot was used to detect the Hippo-YAP/TAZ signaling pathway， including YAP， phosphorylated YAP （p-YAP）， phosphorylated MST1/2 （p-MST1/2）， phosphorylated TAZ （p-TAZ） and apoptosis-related proteins， such as Cleaved Caspase 3， P53， Bcl-2 and Bax. ResultsCompared with sham surgery group， the rats in the model group showed significant degenerative changes in the intervertebral disc. The levels of aggrecan， Bcl-2， and YAP proteins in the nucleus pulposus decreased， while the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and the apoptosis rate increased （P < 0.01）. Compared with the model group， the drug intervention groups showed partial recovery in intervertebral disc degeneration. The levels of aggrecan， Bcl-2， and YAP proteins increased， while the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and the apoptosis rate decreased （P＜0.05 or P＜0.01）. The high-dose ZGD group showed more significant recovery in intervertebral disc degeneration compared to the low-dose ZGD group， with a decrease in the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and apoptosis rate， and an increase in the levels of aggrecan， Bcl-2， and YAP proteins （P＜0.05 or P＜0.01）. Compared with the high-dose ZGD group， the high-dose ZGD + inhibitor group showed a reduced recovery in intervertebral disc degeneration， with an increase in the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and apoptosis rate， and a decrease in the levels of aggrecan， Bcl-2， and YAP proteins （P＜0.05 or P＜0.01）. ConclusionZGD may delay intervertebral disc degeneration by inhibiting the phosphorylation of YAP in the nucleus pulposus， maintaining the function of the Hippo-YAP/TAZ signaling pathway， and reducing apoptosis of nucleus pulposus cells.