ObjectiveTo investigate the mechanism by which 2，3，5，4'-tetrahydroxyldiphenylethylene-2-O-glucoside （THSG） mitigates cerebral ischemia/reperfusion （CI/R） injury by regulating mitochondrial calcium overload and promoting mitophagy. MethodsSixty male SD rats were randomized into sham， model， SAS （40 mg·kg^-1）， and low-， medium- and high-dose （10， 20， 40 mg·kg^-1， respectively） THSG groups， with 10 rats in each group. The middle cerebral artery occlusion/reperfusion （MCAO/R） model was established by the modified Longa suture method. An oxygen-glucose deprivation/reoxygenation （OGD/R） model was constructed in PC12 cells. Neurological deficits were assessed via Zea Longa scoring， and cerebral infarct volume was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Structural and functional changes of cortical neurons in MCAO/R rats were assessed by hematoxylin-eosin and Nissl staining. PC12 cell viability was detected by cell counting kit-8 （CCK-8） assay， and mitochondrial calcium levels were quantified by Rhod-2 AM. Immunofluorescence was used to detect co-localization of PTEN-induced kinase 1 （PINK1） and leucine zipper/EF-hand-containing transmembrane protein 1 （LETM1） in neurons. Transmission electron microscopy （TEM） was employed to observe mitochondrial morphology in neurons. Western blot was employed to analyze the expression of translocase of outer mitochondrial membrane 20 （TOMM20）， autophagy-associated protein p62， microtubule-associated protein light chain 3 （LC3）， cysteinyl aspartate-specific proteinase-9 （Caspase-9）， B-cell lymphoma 2-associated protein X （Bax）， and cytochrome C （Cyt C）. ResultsCompared with the sham group， the model group exhibited increased infarct volume （P<0.01） and neurological deficit scores （P<0.01）， neuronal structure was disrupted with reduced Nissl bodies. （P<0.01）， mitochondrial swelling/fragmentation， decreased PINK1/LETM1 co-localization （P<0.01）， upregulated protein levels of LC3Ⅱ/LC3Ⅰ， TOMM20， Caspase-9， Bax， and Cyt C （P<0.01）， downregulated protein level of p62 （P<0.05）， weakened PC12 viability （P<0.01）， and elevated mitochondrial calcium level （P<0.01）. Compared with the model group， THSG and SAS groups showed reduced infarct volumes （P<0.05，P<0.01） and neurological deficit scores （P<0.05，P<0.01）， mitigated mitochondrial damage， and increased PINK1/LETM1 co-localization （P<0.01）. Medium/high-dose THSG and SAS alleviated the neurological damage， increased Nissl bodies （P<0.05，P<0.01）， downregulated the protein levels of p62， TOMM20， Caspase-9， Bax， and Cyt C （P<0.05，P<0.01）， and elevated the LC3Ⅱ/LC3Ⅰ level （P<0.05，P<0.01）. High-dose THSG enhanced PC12 cell viability （P<0.01）， increased PINK1/LETM1 co-localization （P<0.01）， and reduced mitochondrial calcium （P<0.01）. ConclusionTHSG may exert the neuroprotective effect on CI/R injury by activating the PINK1-LETM1 signaling pathway， reducing the mitochondrial calcium overload， and promoting mitophagy.

ObjectiveTo investigate the mechanism by which 2，3，5，4'-tetrahydroxyldiphenylethylene-2-O-glucoside （THSG） mitigates cerebral ischemia/reperfusion （CI/R） injury by regulating mitochondrial calcium overload and promoting mitophagy. MethodsSixty male SD rats were randomized into sham， model， SAS （40 mg·kg^-1）， and low-， medium- and high-dose （10， 20， 40 mg·kg^-1， respectively） THSG groups， with 10 rats in each group. The middle cerebral artery occlusion/reperfusion （MCAO/R） model was established by the modified Longa suture method. An oxygen-glucose deprivation/reoxygenation （OGD/R） model was constructed in PC12 cells. Neurological deficits were assessed via Zea Longa scoring， and cerebral infarct volume was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Structural and functional changes of cortical neurons in MCAO/R rats were assessed by hematoxylin-eosin and Nissl staining. PC12 cell viability was detected by cell counting kit-8 （CCK-8） assay， and mitochondrial calcium levels were quantified by Rhod-2 AM. Immunofluorescence was used to detect co-localization of PTEN-induced kinase 1 （PINK1） and leucine zipper/EF-hand-containing transmembrane protein 1 （LETM1） in neurons. Transmission electron microscopy （TEM） was employed to observe mitochondrial morphology in neurons. Western blot was employed to analyze the expression of translocase of outer mitochondrial membrane 20 （TOMM20）， autophagy-associated protein p62， microtubule-associated protein light chain 3 （LC3）， cysteinyl aspartate-specific proteinase-9 （Caspase-9）， B-cell lymphoma 2-associated protein X （Bax）， and cytochrome C （Cyt C）. ResultsCompared with the sham group， the model group exhibited increased infarct volume （P<0.01） and neurological deficit scores （P<0.01）， neuronal structure was disrupted with reduced Nissl bodies. （P<0.01）， mitochondrial swelling/fragmentation， decreased PINK1/LETM1 co-localization （P<0.01）， upregulated protein levels of LC3Ⅱ/LC3Ⅰ， TOMM20， Caspase-9， Bax， and Cyt C （P<0.01）， downregulated protein level of p62 （P<0.05）， weakened PC12 viability （P<0.01）， and elevated mitochondrial calcium level （P<0.01）. Compared with the model group， THSG and SAS groups showed reduced infarct volumes （P<0.05，P<0.01） and neurological deficit scores （P<0.05，P<0.01）， mitigated mitochondrial damage， and increased PINK1/LETM1 co-localization （P<0.01）. Medium/high-dose THSG and SAS alleviated the neurological damage， increased Nissl bodies （P<0.05，P<0.01）， downregulated the protein levels of p62， TOMM20， Caspase-9， Bax， and Cyt C （P<0.05，P<0.01）， and elevated the LC3Ⅱ/LC3Ⅰ level （P<0.05，P<0.01）. High-dose THSG enhanced PC12 cell viability （P<0.01）， increased PINK1/LETM1 co-localization （P<0.01）， and reduced mitochondrial calcium （P<0.01）. ConclusionTHSG may exert the neuroprotective effect on CI/R injury by activating the PINK1-LETM1 signaling pathway， reducing the mitochondrial calcium overload， and promoting mitophagy.

OBJECTIVE: To explore the feasibility and effectiveness of suture anchor double-pulley technique combined with suture three-dimensional binding via bone tunnel technique for avulsion fractures of the inferior pole of the patella. METHODS: A clinical data of 38 patients with avulsion fractures of the inferior pole of the patella, who met the selective criteria and were admitted between September 2021 and April 2023, was retrospectively analyzed. The fractures were treated with suture anchor double-pulley technique combined with suture three-dimensional binding via bone tunnel technique in 18 cases (group A) and steel wire tension-band fixation in 20 cases (group B). There was no significant difference in terms of age, gender, cause of fracture, side of fracture, and disease duration between the two groups ( P>0.05). The length of incision, operation time, occurrence of complications, the range of motion of knee joint, and Böstman score of knee joint at last follow-up were recorded. The fracture healing was evaluated through X-ray films and the time of fracture healing was recorded. RESULTS: All incisions healed by first intention. The length of incision was significantly shorter in group A than in group B ( P<0.05). There was no significant difference in the operation time between the two groups ( P>0.05). All patients were followed up 12-24 months (mean, 16.1 months). X-ray films showed that all fractures healed and there was no significant difference in the healing time between the two groups ( P>0.05). At last follow-up, the range of motion and Böstman score of the knee joint in group A were significantly better than those in group B ( P<0.05). During follow-up, 1 patient (5.6%) in group A had one anchor mild prolapse and 3 patients (15.0%) occured internal fixation irritation in group B. But there was no significant difference in the incidence of complications between the two groups ( P>0.05). CONCLUSION For the avulsion fractures of the inferior pole of the patella, the suture anchor double-pulley technique combined with suture three-dimensional binding via bone tunnel technique has advantages of reliable fixation, small incision, avoidance of secondary operation to remove internal fixator, and fewer complications, with definite effectiveness.
Humans ; Male ; Female ; Patella/surgery* ; Suture Anchors ; Fracture Fixation, Internal/instrumentation* ; Adult ; Retrospective Studies ; Middle Aged ; Fractures, Avulsion/surgery* ; Treatment Outcome ; Young Adult ; Range of Motion, Articular ; Fracture Healing ; Adolescent ; Suture Techniques ; Knee Joint/physiopathology*

OBJECTIVE: To compare the effectiveness of using a composite loop plate to reconstruct the coracoclavicular ligament around the coracoid process and using a clavicular hook plate for fixation in treatment of Rockwood type Ⅲ acute acromioclavicular joint dislocation. METHODS: A retrospective analysis was conducted on the clinical data of 60 patients with Rockwood type Ⅲ acute acromioclavicular joint dislocation who were admitted between June 2022 and September 2023 and met the selection criteria. Among them, 30 patients were treated with the composite loop plate to reconstruct the coracoclavicular ligament around the coracoid process (loop plate group) and 30 with clavicular hook plate fixation (hook plate group). There was no significant difference in baseline data between the two groups ( P>0.05), including gender, age, injured side, cause of injury, disease duration, preoperative visual analogue scale (VAS) score for pain, and Constant-Murley score. The incision length, operation time, length of hospital stay, and the occurrence of complications during follow-up were recorded. The Constant-Murley score and VAS score were used to evaluate shoulder joint function and pain, and the differences (change values) of the indicators between before operation and at 6 months after operation were calculated for inter-group comparison. In the loop plate group, the coracoclavicular distance (CCD) on the anteroposterior X-ray films of the acromioclavicular joint was measured at 1 day and 6 months after operation to assess the loss of acromioclavicular joint reduction. RESULTS: The incision length of the loop plate group was significantly shorter than that of the hook plate group ( P<0.05). There was no significant difference in the operation time and the length of hospital stay between the two groups ( P>0.05). All incisions healed by first intention after operation. All patients were followed up 12-18 months (mean, 16.3 months). There was no significant difference in the follow-up time between groups ( P>0.05). The Constant-Murley scores and VAS scores of both groups significantly improved at 6 months after operation when compared with those before operation ( P<0.05); the differences in the change values of the two indicators between groups were significant ( P<0.05). The CCD of the loop plate group were (10.40±0.83) mm at 1 day and (10.70±0.68) mm at 6 months and no repositioning loss was observed. Three cases in the hook plate group had residual shoulder joint pain after operation. The difference in the accidence of complications between groups was not significant ( P>0.05). CONCLUSION For Rockwood type Ⅲ acute acromioclavicular joint dislocation, compared with the clavicular hook plate fixation, the composite loop plate for reconstructing the coracoclavicular ligament around the coracoid process has the advantages of simple operation, safety, minimally invasive, good functional recovery, and fewer complications. Moreover, it avoids the need for a second surgery to remove the internal fixation device, and the patient acceptance and satisfaction are higher.
Humans ; Acromioclavicular Joint/surgery* ; Bone Plates ; Male ; Retrospective Studies ; Female ; Adult ; Ligaments, Articular/injuries* ; Joint Dislocations/surgery* ; Coracoid Process/injuries* ; Treatment Outcome ; Middle Aged ; Plastic Surgery Procedures/instrumentation* ; Fracture Fixation, Internal/instrumentation* ; Young Adult ; Clavicle/surgery*

Objective To investigate the molecular mechanisms by which SOX7 regulates the SHP-2/Wnt/β-cate-nin/ROS pathway,affecting the proliferation,invasion,and migration of colorectal cancer cells.Methods Twenty nude mice with subcutaneously transplanted tumor models were randomly divided into four groups:SOX7 NC(n=5),SOX mimic(n=5),SOX7 NC+PHPS1(n=5),and SOX7 mimic+PHPS1(n=5)to observe tumor growth.Human colorectal cancer cell line SW480 cells were transfected via lipofection and divided into six groups:SOX7 NC,SOX7 mimic,SOX7 NC+H2 O2,SOX7 mimic+H2O2,SOX7 NC+PHPS1,and SOX7 mimic+PHPS1.The ex-pression of SHP-2/Wnt/β-catenin/ROS pathway-related proteins in SW480 cells of each group was detected by Western blot.The invasion and migration capabilities of SW480 cells were assessed through scratch and Transwell invasion assays,while cell proliferation was evaluated using CCK-8.Results In vivo experiments demonstrated that tumors in the SOX7 mimic group were significantly smaller than those in the SOX7 NC group(P＜0.01).Tumors treated with PHPS1 intervention exhibited a significant increase in volume.There was no statistical significance in the difference in tumor volume between the SOX7 mimic+PHPS1 group and the SOX7 NC+PHPS1 group.In vitro experiments revealed that SOX7 mimic inhibited the expression of Wnt,β-catenin,NOX2,NOX4,PI3K,P-PI3K,AKT,P-AKT proteins(P＜0.01),and promoted the expression of p-SHP-2 protein(P＜0.01).The addition of hydrogen peroxide and SHP-2 inhibitor reversed the effects of SOX7 on SW480 cells(P＜0.05),and significantly promoted the expression levels of Wnt,β-catenin,NOX2,NOX4,PI3K,P-PI3K,AKT,P-AKT proteins,with no sig-nificant difference,while significantly reducing the expression levels of SHP-2,p-SHP-2 proteins,with no significant difference.PHPS1 inhibited the expression of SHP-2,p-SHP-2 proteins(P＜0.05)and upregulated the expression of Wnt,β-catenin,NOX2,NOX4,PI3K,P-PI3K,AKT,P-AKT proteins(P＜0.05).Scratch,Transwell invasion and migration assays,and CCK-8 experiments indicated that SOX7 suppressed the migration,invasion,and proliferation of SW480 cells through oxidative stress and the SHP-2 pathway(P＜0.01),while H2O2 and PHPS1 intervention promoted the migration,invasion,and proliferation of SW480 cells(P＜0.05).Conclusion SOX7 can suppress the proliferation,invasion,and migration of colorectal cancer by targeting the SHP-2/Wnt/β-catenin/ROS pathway.

Objective To investigate the effect of Zuogui Jiangtang Jieyu Formula(左归降糖解郁方,ZJJF)on hippocampal neuron apoptosis in diabetic rats with depression and to ascertain whether its mechanism involves the regulation of JNK signaling pathway. Methods(i)A total of 72 specific pathogen-free(SPF)grade male Sprague Dawley(SD)rats were randomly divided into six groups,with 12 rats in each group:control,model,metformin(Met,0.18 g/kg)+fluoxetine(Flu,1.8 mg/kg),and the high-,medium-,and low-ZJJF dosages(ZJJF-H,20.52 g/kg;ZJJF-M,10.26 g/kg;ZJJF-L,5.13 g/kg)groups.All groups except control group were injected once via the tail vein with streptozotocin(STZ,38 mg/kg)combined with 28 d of chronic unpredictable mild stress(CUMS)to establish diabetic rat models with de-pression.During the CUMS modeling period,treatments were administered via gavage,with control and model groups receiving an equivalent volume of distilled water for 28 d.The effi-cacy of ZJJF in reducing blood sugar and alleviating depression was evaluated by measuring fasting blood glucose,insulin,and glycated hemoglobin levels,along with behavioral assess-ments,including the open field test(OFT),forced swim test(FST),and sucrose preference test(SPT).Hippocampal tissue damage and neuronal apoptosis were evaluated using hema-toxylin-eosin(HE)staining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining.Apoptosis-related proteins Bax,Bcl-2,caspase-3,and the ex-pression levels of JNK/Elk-1/c-fos signaling pathway were detected using Western blot and real-time quantitative polymerase chain reaction(RT-qPCR).(ii)To further elucidate the role of JNK signaling pathway in hippocampal neuronal apoptosis and the pharmacological ef-fects of ZJJF,an additional 50 SPF grade male SD rats were randomly divided into five groups,with 10 rats in each group:control,model,SP600125(SP6,a JNK antagonist,10 mg/kg),ZJJF(20.52 g/kg),and ZJJF(20.52 g/kg)+Anisomycin(Aniso,a JNK agonist,15 mg/kg)groups.Ex-cept for control group,all groups were established as diabetic rat models with depression,and treatments were administered via gavage for ZJJF and intraperitoneal injection for SP6 and Aniso for 28 d during the CUMS modeling period.Behavioral changes in rats were evaluated through the OFT,FST,and SPT,and hippocampal neuron damage and apoptosis were ob-served using HE staining,Nissl staining,TUNEL staining,and transmission electron mi-croscopy(TEM).Changes in apoptosis-related proteins and JNK signaling pathway in the hippocampal tissues of rats were also analyzed. Results(i)ZJJF significantly reduced the high blood glucose,insulin,and glycated he-moglobin levels in model rats(P<0.01).It increased autonomous activity and decreased de-spair-like behaviors(P<0.01),improved the pathological damage of hippocampal neurons,increased the number of neuronal nuclei(P<0.01),and reduced the number of mechanocytes,vacuolar cells,and apoptotic neurons(P<0.05,P<0.01,and P<0.01,respec-tively).ZJJF down-regulated the expression levels of pro-apoptotic proteins Bax and caspase-3(P<0.01),up-regulated the anti-apoptotic protein Bcl-2(P<0.01),and significantly inhibit-ed the overexpression of phosphorylated JNK(p-JNK),Elk-1,and c-fos(P<0.01).(ii)SP6 in-creased autonomous activity and reduced despair time in model rats(P<0.05),although it had no significant effects on sucrose preference(P>0.05).It increased the number of Nissl bodies in hippocampal neurons(P<0.01),reduced the protein expression levels of Bax(P<0.01)and caspase-3(P<0.05),and decreased the number of apoptotic neurons(P<0.05).SP6 also increased the expression level of Bcl-2(P<0.01),and inhibited the high expression levels of p-JNK,Elk-1,and c-fos(P<0.01,P<0.01,and P<0.05,respectively),suggesting that hip-pocampal neuronal apoptosis in diabetic rats with depression is associated with abnormal ac-tivation of JNK signaling pathway.Compared with ZJJF group,ZJJF+Aniso group showed a decrease in sucrose preference(P<0.05)and an increase in despair time(P<0.01)with more notable hippocampal neuronal damage.This group also exhibited a decrease in expression level(P<0.01)Bcl-2 and an increase in expression levels of Bax,caspase-3,p-JNK,Elk-1,and c-fos(P<0.01,P<0.05,P<0.05,P<0.01,and P<0.05,respectively),indicating that the antidepressant effects of ZJJF,its improvement of neuronal apoptosis,and regulation of JNK signaling molecules could all be reversed by a specific JNK agonist. Conclusion ZJJF exerts a significant hypoglycemic effect and ameliorates the apoptosis of hippocampal neurons by inhibiting the activation of JNK signaling pathway,which is a promising formula for the treatment of diabetic depression in clinical settings.

Objective:To investigate the effect of different temperature and time treatment of Xiaoying Fang on wound healing after mixed hemorrhoid surgery.Methods:A total of 160 patients with mixed hemorrhoids who underwent surgical treatment in Fuyang Hospital of Traditional Chinese Medicine from September 2020 to October 2023 were prospectively selected as the study objects, according to the random number table method and the patients who withdrew were excluded, and 157 cases were finally enrolled, and the patients were divided into observation group 1 (39 cases), observation group 2 (40 cases), observation group 3 (38 cases) and observation group 4 (40 cases). The four groups were all treated with Xiaoying Fang hip bath after surgery. The temperature and time of hip bath in group 1 were observed at 38 - 41 ℃ for 10 min, group 2 at 38 - 41 ℃ for 15 min, group 3 at 42 - 45 ℃ for 10 min, and group 4 at 42 - 45 ℃ for 15 min. Wound pain, edema, wound granulation growth and wound healing were compared between the four groups after treatment.Results:At the 3rd and 5th day of treatment, the visual analogue scale (VAS) scores in the observation group 1 and observation group 2 were lower than those in the observation group 3 and observation group 4: (4.33 ± 1.19), (4.43 ± 1.13) scores vs. (5.15 ± 1.05), (5.40 ± 1.26) scores; (2.46 ± 1.25), (2.63 ± 0.54) scores vs. (3.61 ± 1.22), (3.85 ± 1.05) scores, there were statistical differences ( P<0.05). The wound edema scores in the observation group 2 were lower than that in the observation group 1, observation group 3 and observation group 4: (1.43 ± 0.50) scores vs. (2.21 ± 0.83), (2.05 ± 0.84), (1.98 ± 0.86) scores; (0.88 ± 0.22) scores vs. (1.38 ± 0.54), (1.00 ± 0.23), (0.93 ± 0.25) scores, there were statistical differences ( P<0.05). On the 3rd, 5th and 7th day of treatment, the wound granulation growth scores in the observation group 2 was lower than that in the observation group 1, observation group 3 and observation group 4: (0.80 ± 0.08) scores vs. (1.36 ± 0.13), (1.42 ± 0.13), (1.53 ± 0.15) scores; (0.65 ± 0.18) scores vs. (1.08 ± 0.17), (1.07 ± 0.21), (1.03 ± 0.10) scores; (0.35 ± 0.08) scores vs. (0.74 ± 0.17), (0.97 ± 0.15), (0.78 ± 0.09) scores, there were statistical differences ( P<0.05). There was significant difference in wound healing among the four groups after operation 7 d ( H = 9.84, P<0.05). The total effective rate of wound healing in the observation group 2 was the highest, which was 92.50% (37/40). Conclusions:The optimal temperature and treatment time of Xiaoying Fang hip bath are 38 - 41 ℃ and 15 min.

Purpose/Significance To explore the application and predictive accuracy of various models in predicting the risk of ather-osclerosis in diabetic patients.Method/Process Based on the biochemical data table from the"Diabetes Complications Warning Dataset"provided by the National Population Health Science Data Center,MATLAB software is used to construct risk prediction models for diabe-tes-induced atherosclerosis.The models are built by using k-nearest neighbors(KNN),decision trees,backpropagation(BP)neural networks,and Naive Bayes algorithms,and which are subjected to comparative analysis.Result/Conclusion In terms of effectiveness,the predictive accuracy of Naive Bayes algorithm is the highest(61.6％),followed by the decision tree model(58.2％),the KNN mod-el(57.7％),and the BP neural network model(55.9％).The results of the confusion matrix and the receiver operating characteristic(ROC)curve indicate that the Naive Bayes model performs best.When comparing the models in terms of effectiveness,performance and stability,the Naive Bayes model is superior.

AIM:To explore the mechanism by which Chaijin-Jieyu-Anshen tablet(CJJY)-medicated serum prevents synaptic injury in rat anterior cingulate cortex(ACC)neurons using an in vitro depression model.METHODS:Cells(astrocytes,microglia and neurons)were isolated from the ACC of SD rats.The isolated cells were characterized by immunofluorescence staining.An in vitro depression model was developed using 1 mg/L lipopolysaccharide(LPS)com-bined with 200 μmol/L corticosterone(CORT).These cells were divided into control group,model group(CORT+LPS),glucocorticoid receptor(GR)blocker(GR-)group(CORT+LPS+RU486),GR agonist(GR+)group(CORT+LPS+dexa-methasone),CX3C chemokine receptor 1(CX3CR1)blocker(CX3-)group(CORT+LPS+AZD8797),CX3CR1 agonist(CX3+)group(CORT+LPS+fractalkine),CJJY group(CORT+LPS+CJJY-medicated serum),CJJY/GR+group(CORT+LPS+CJJY-medicated serum+dexamethasone),and CJJY/CX3+group(CORT+LPS+CJJY-mediated serum+fractalkine).The morphological characteristics of all ACC cells were observed by high-content analysis.The levels of neuroendocrine-related factors,adrenocorticotropic hormone(ACTH),corticotropin-releasing hormone(CRH)and CORT,and neuroin-flammatory mediators,tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-6 and glutamate(Glu),in the cell supernatants were quantified by ELISA.Immunofluorescence staining was used to analyze the protein expression of GR and vesicular glutamate transporter 1(VGluT1)in astrocytes,as well as CX3CR1 and adenosine A2A receptor(A2AR)in microglia.Immunofluorescence staining with Nissl and β-tubulin was performed to evaluate synaptic damage in ACC neurons.RESULTS:In an in vitro model of depression,CJJY-medicated serum prevented morphological damage to ACC neurons,microglia and astrocytes.Moreover,CJJY-medicated serum reversed abnormal increases in the levels of ACTH,CRH,CORT,TNF-α,IL-1β,IL-6 and Glu in cell supernatants(P<0.05 or P<0.01).It was also found that CJJY-medi-cated serum reduced abnormal expression of GR,VGluT1,CX3CR1 and A2AR(P<0.05 or P<0.01),alleviating damage to the neuronal dendrites and dendritic spines of ACC neurons.CONCLUSION:The CJJY-medicated serum regulates GR/CX3CR1 double signaling in glia,and attenuates rat ACC neuronal synaptic damage in an in vitro depression model,indicating that CJJY-medicated serum controls depression by affecting the GR/CX3CR1 double signaling.

ObjectiveTo investigate the mechanism of Baihuan Xiaoyao Decoction （Xiaoyaosan added with Lilii Bulbus and Albiziae Cortex） in alleviating depression-like behaviors of juvenile rats by regulating the polarization of microglia. MethodSixty juvenile SD rats were randomized into normal control， model， fluoxetine， and low-， medium-， and high-dose （5.36， 10.71， 21.42 g·kg^-1， respectively） Baihuan Xiaoyao decoction groups. The rat model of juvenile depression was established by chronic unpredictable mild stress （CUMS）. The sucrose preference test （SPT） was carried out to examine the sucrose preference of rats. Forced swimming test （FST） was carried out to measure the immobility time of rats. The open field test （OFT） was conducted to measure the total distance， the central distance， the number of horizontal crossings， and the frequency of rearing. Morris water maze （MWM） was used to measure the escape latency and the number of crossing the platform. The immunofluorescence assay was employed to detect the expression of inducible nitric oxide synthase （iNOS， the polarization marker of M1 microglia） and CD206 （the polarization marker of M2 microglia）. Real-time polymerase chain reaction was employed to determine the mRNA levels of iNOS， CD206， pro-inflammatory cytokines ［tumor necrosis factor （TNF）-α， interleukin （IL）-1β， and IL-6］ and anti-inflammatory cytokines （IL-4 and IL-10） in the hippocampus. Western blotting was employed to determine the protein levels of iNOS and CD206 in the hippocampus. The levels of IL-4 and IL-6 in the hippocampus were detected by enzyme-linked immunosorbent assay. ResultCompared with the normal control group， the model rats showed a reduction in sucrose preference （P<0.05）， an increase in immobility time （P<0.05）， decreased motor and exploratory behaviors （P<0.05）， and weakened learning and spatial memory （P<0.05）. In addition， the model rats showed up-regulated mRNA and protein levels of iNOS and mRNA levels of IL-1β， IL-6， and TNF-α （P<0.05）. Compared with the model group， Baihuan Xiaoyao decoction increased the sucrose preference value （P<0.05）， shortened the immobility time （P<0.01）， increased the motor and exploratory behaviors （P<0.05）， and improved the learning and spatial memory （P<0.01）. Furthermore， the decoction down-regulated the positive expression and protein level of iNOS， lowered the levels of TNF-α， IL-1β， and IL-6 （P<0.01）， promoted the positive expression of CD206， and elevated the levels of IL-4 and IL-10 （P<0.01） in the hippocampus of the high dose group. Moreover， the high-dose Baihuan Xiaoyao decoction group had higher sucrose preference value （P<0.01）， shorter immobility time （P<0.01）， longer central distance （P<0.01）， stronger learning and spatial memory （P<0.01）， higher positive expression and protein level of iNOS （P<0.01）， lower levels of TNF-α， IL-1β， and IL-6 （P<0.05， P<0.01）， lower positive expression and mRNA level of iNOS （P<0.05）， and higher levels of IL-4 and IL-10 （P<0.05， P<0.01） than the fluoxetine group. ConclusionBaihuan Xiaoyao decoction can improve the depression-like behavior of juvenile rats by inhibiting the M1 polarization and promoting the M2 polarization of microglia in the hippocampus.