Regulatory Effect of Wenyang Prescription， Jieyu Prescription， and Wenyang Jieyu Prescription on Pain Sensitivity and Depression-like Behaviors in Mice Induced by Maternal Separation and Chronic Neuropathic Pain / 中国实验方剂学杂志

ObjectiveTo observe the behavioral and pain threshold alterations， as well as the changes in indexes related to depression and pain in the serum and central system in mice stressed by maternal separation and chronic neuropathic pain， and explore the underlying mechanism of Wenyang prescription （WY）， Jieyu prescription （JY）， and Wenyang Jieyu prescription （WYJY） in improving depression and pain sensitivity. MethodThe birth date of mice was recorded as PD0. After birth， the mice were divided into a blank group and an experimental group. The neonatal mice in the experimental group underwent maternal separation in PD5-14 at 8 h·d-1. After ablactation， the mice were divided into a maternal separation group， a WY group （Erxian decoction， 5.84 g·kg-1）， a JY group （Xiaoyaosan， 12.00 g·kg-1）， a WYJY group （16.68 g·kg-1）， and a fluoxetine group （2.60 mg·kg-1）， with 15 mice in each group. Meanwhile， 15 male mice of the same age without maternal separation were assigned to the normal control group. Mice in the blank group and the maternal separation group were fed on a regular chow diet in PD21-PD90， while the remaining groups were fed on the corresponding drugs. In PD91， sciatic nerve ligation was performed to induce a model of maternal separation and chronic neuropathic pain. The open field test was used to observe the depression-like behaviors of mice in each group， and the mechanical and temperature pain thresholds were measured to detect the pain sensitivity of mice in each group. The serum levels of corticosterone （CORT）， substance P， and β-endorphin （β-EP） were determined by enzyme-linked immunosorbent assay （ELISA）， and the expression of the glucocorticoid receptor （GR） in the amygdala and β-EP protein in the hypothalamus was detected by immunohistochemistry. The mRNA expression levels of amygdala GR gene （Nr3c1）， FK506 binding protein 5 gene （FKBP5）， metabolic glutamate receptor 5 gene （GRM5）， and brain-derived neurotrophic factor （BDNF） were detected by real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. ResultCompared with the blank group， the maternal separation group showed reduced stay time and total distance traveled in the 5-min open field test （P<0.01）， reduced mechanical pain threshold （P<0.01）， increased serum CORT and β-EP （P<0.01）， declining FKBP5 mRNA expression （P<0.01）， and increased hypothalamic β-EP expression （P<0.05）. Compared with the maternal separation group， the groups with drug intervention showed prolonged stay time （P<0.05， P<0.01） and up-regulated pain thresholds to different degrees. The total distance traveled in the 5-min open field test increased in the WY group， the WYJY group， and the fluoxetine group （P<0.05， P<0.01）. The JY group showed decreased serum CORT （P<0.01）， reduced β-EP ， and increased BDNF mRNA （P<0.01）. Nr3c1 and GRM5 mRNA decreased in the WY group （P<0.05， P<0.01）. The WYJY group showed decreased serum CORT （P<0.05）and decreased Nr3c1， GRM5， and BDNF mRNA （P<0.05， P<0.01）. The levels of β-EP expression were elevated to different degrees in the groups with drug intervention， but the differences were not significant. The levels of GR expression in the WY group， the JY group， and the WYJY group increased （P<0.05）. ConclusionWYJY can inhibit central pain sensitization and regulate hypothalamic-pituitary-adrenal gland （HPA） axis function by enhancing the expression of GR in the amygdala and inhibiting neuroplasticity and excitability in the amygdala to relieve depression-like behaviors and improve somatic hyperalgesia.

Clinical Study of Chaihu Shugansan Combined with Abdominal Acupuncture on Depression Caused by Chronic Pain / 中国实验方剂学杂志

Objective:To investigate the clinical effect of Chaihu Shugansan combined with abdominal acupuncture on depression caused by chronic pain，and to explore its mechanism. Method:A total of 97 patients with depression caused by chronic pain were randomly divided into control group （49 cases） and observation group （48 cases）. Patients in both groups received routine western medicine treatment，including necessary psychological intervention and taking paroxetine. Control groupobservation groupcontrol group Patients in control group were treated with Xiaoyaowan，and patients in observation group were treated with Chaihu Shugansan combined with abdominal acupuncture. Both groups were treated for 6 weeks. The levels of serum neurotransmitters，cytokines and Hamilton depression rating scale（HAMD） before and after treatment were compared between two groups<bold>.</bold> Result:There was no significant difference in HAMD scores of the two groups before treatment and the HAMD scores of two groups after treatment were significantly lower than those before treatment （<italic>P</italic><0.05），and the HAMD scores in observation group were significantly lower than those in the control group （<italic>P</italic><0.05）. There was no significant difference in the levels of serum 5-hydroxytryptamine （5-HT），norepinephrine （NE），and brain-derived neurotrophic factor （BDNF） between two groups before treatment. After treatment，the levels of serum 5-HT，NE，and BDNF in two groups were significantly higher than those before treatment （<italic>P</italic><0.05），and the levels in observation group were significantly higher than those in control group （<italic>P</italic><0.05）. There was no significant difference in the levels of serum interleukin-6 （IL-6），interleukin-1<italic>β</italic> （IL-1<italic>β</italic>） and tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>） before treatment. After treatment，the levels of serum IL-6，IL-1<italic>β</italic> and TNF-<italic>α</italic> were significantly lower than those before treatment （<italic>P</italic><0.05），and the levels in observation group were significantly lower than those in control group （<italic>P</italic><0.05）. Conclusion:On the basis of psychological intervention and paroxetine administration，the combination of Chaihu Shugansan and abdominal acupuncture exerts their respective advantages. It treats both symptoms and root causes of depression，relieves the degree of depression，reduces the classification of depression，and regulates the levels of neurotransmitter and cellular inflammatory factors，and inhibits inflammatory response. The clinical effect is significant.

Changes of Microglia in Hippocampus of Mice Induced by Maternal Separation with Restraint Stress and Regulatory Effect of Wenyang Jieyu Prescription / 中国实验方剂学杂志

Objective:To observe the activation of microglia in hippocampus of depressed and anxious mice induced by maternal separation with acute restraint stress and the expression of interleukin-1<italic>β</italic>（IL-1<italic>β</italic>），interleukin-6（IL-6），tumor necrosis factor-<italic>α</italic>（TNF-<italic>α</italic>）， investigating the mechanism of Wenyang Jieyu prescription in treating anxiety and depression. Method:Eighty four male C57BL offspring were randomly divided into control group， acute restraint stress group and model group on postnatal day 0（PD0）. Maternal separation combined with acute restraint stress was used to prepare anxious and depressed model mice， dividing the model mice into model group， Wenyang， Jieyu， Wenyang Jieyu and fluoxetine group according to random number table method. During the period of PD21-PD90， the control， acute restraint stress and model mice were fed with normal diet， with the other groups fed with corresponding medicine mixed diet. The Wenyang， Jieyu and Wenyang Jieyu groups were given 5.85， 12.03 and 16.71 g·kg^-1·d^-1 respectively. The fluoxetine group was given 2.60 mg·kg^-1·d^-1. Open field， zero maze test and social interaction tests were used to evaluate the anxiety and depression of model mice. The expression of Iba-1 in hippocampal microglia was detected by immunohistochemistry（IHC）. The mRNA expression of IL-1<italic>β</italic>， IL-6， TNF-<italic>α</italic>， Iba-1 and glucocorticoid receptor（GR）were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Result:Compared with the control group， total movement distance and time spent in central zone in 5 min of the model mice significantly decreased（<italic>P</italic><0.01）， time spent in opened arm and total movement distance decreased significantly（<italic>P</italic><0.05，<italic>P</italic><0.01）， investigation time during testing and training increased significantly（<italic>P</italic><0.01）. The expression of Iba-1 protein and mRNA，IL-1<italic>β</italic>，IL-6，TNF-<italic>α</italic> mRNA significantly increased（<italic>P</italic><0.01）， the expression levels of GR mRNA significantly decreased（<italic>P</italic><0.01）. The result of IHC staining showed that microglia were over activated. Compared with the model group， total movement distance and time spent in central zone in 5 min of mice in the Wenyang Jieyu and fluoxetine group significantly increased（<italic>P</italic><0.05，<italic>P</italic><0.01）.Time spent in opened arm significantly increased（<italic>P</italic><0.01）. Investigation time during testing and training significantly decreased（<italic>P</italic><0.05，<italic>P</italic><0.01）. The expression of Iba-1 protein and mRNA，IL-1<italic>β</italic>，IL-6，TNF-<italic>α</italic> mRNA significantly decreased（<italic>P</italic><0.05，<italic>P</italic><0.01）. The expression of GR mRNA increased significantly（<italic>P</italic><0.05，<italic>P</italic><0.01）. IHC staining showed the microglia recovered. Time spent in opened arm of mice in the Wenyang group and Jieyu group significantly increased （<italic>P</italic><0.01）， time spent investigating during testing decreased significantly （<italic>P</italic><0.05）， the expression levels of Iba-1 protein and mRNA，IL-6 mRNA significantly decreased （<italic>P</italic><0.05，<italic>P</italic><0.01）. The expression of GR mRNA of mice in the Wenyang group significantly increased （<italic>P</italic><0.05）， the expression of TNF-<italic>α </italic>mRNA significantly decreased （<italic>P</italic><0.05）. Total movement distance of mice in the Jieyu group increased significantly （<italic>P</italic><0.01）， time spent investigating during training decreased significantly （<italic>P</italic><0.05），the expression level of IL-1<italic>β </italic>mRNA significantly decreased （<italic>P</italic><0.05）. IHC staining showed that microglia recovered partly in both groups. Conclusion:The comprehensive curative effect and pharmacological action of Wenyang Jieyu prescription were better than Wenyang prescription and Jieyu prescription. Wenyang Jieyu prescription can treat anxiety and depression in maternal separation and acute restraint stress mice， its possible mechanism may be related to the decreased activation of microglia， down-regulation of IL-1<italic>β</italic>，IL-6，TNF-<italic>α</italic> expression and up-regulation of GR expression.

Effect of Wenyang， Jieyu， and Wenyang Jieyu Prescriptions on Hippocampal Microglia of Mice with Depression-Like Behavior Induced by Secondary LPS Exposure / 中国实验方剂学杂志

Objective:To observe the activation of microglia and the expression of inflammatory factors in hippocampus of mice with depression-like behavior after mother-infant separation （MS） combined with lipopolysaccharide （LPS） stress， and to explore the possible anti-depression mechanisms of Wenyang （WY）， Jieyu （JY）， and Wenyang Jieyu （WYJY） prescriptions from the perspective of warming Yang and relieving depression. Method:Seventy offspring mice were randomly divided into a normal group （<italic>n</italic>=10）， a LPS stress group （<italic>n</italic>=10）， and a modeling group （<italic>n</italic>=50）. After undergoing 8 h·d^-1mother-infant separation during postnatal day 5 （PD₅）–PD₁₄， mice in the modeling group were further divided into the MS + LPS group， WY group， JY group， WYJY group， and fluoxetine （FLU） group， with 10 in each group. The birth date of the offspring mice was recorded as PD₀. The mice in the normal， LPS， and MS + LPS groups were fed a normal diet during PD₂₁–PD₉₀， while those in the other groups were treated with the mixtures of corresponding drugs and feed， followed by seven-day intraperitoneal injection of LPS since PD₉₁ for inducing depression. The depression-like behavior of mice in each group was detected in the open-field， O-maze， and social interaction tests. The protein expression of microglia-specific ionized calcium-binding adaptor molecule 1 （Iba-1） in the hippocampus was assayed by immunohistochemistry， and the mRNA expression of interleukin-1<italic>β </italic>（IL-1<italic>β</italic>）， interleukin-6 （IL-6）， tumor necrosis factor-<italic>α </italic>（TNF-<italic>α</italic>）， Iba-1， and glucocorticoid receptor （GR） by real-time fluorescence quantitative PCR （Real-time PCR）. Result:Compared with the normal group， the LPS group exhibited significantly reduced residence time at the central area within 5 min （<italic>P</italic><0.01） and shortened total exercise distance （<italic>P</italic><0.01）. In the MS + LPS group， the open-arm activity time and the total activity distance decreased significantly （<italic>P</italic><0.01， <italic>P</italic><0.05）， whereas the training， discrimination and exploration time increased significantly （<italic>P</italic><0.01）. The expression of Iba-1 in hippocampal CA1 region of mice in the LPS and MS + LPS groups was remarkably elevated （<italic>P</italic><0.01）. Compared with the LPS group， the MS + LPS group displayed significantly prolonged distance of 5-min exercise （<italic>P</italic><0.05）， increased training， discrimination and exploration time （<italic>P</italic><0.05， <italic>P</italic><0.01）， and up-regulated Iba-1 expression in hippocampal CA1 area （<italic>P</italic><0.01）. As revealed by comparison with the MS + LPS group， both the total 5-min exercise distance （<italic>P</italic><0.01） and the training and discrimination time （<italic>P</italic><0.01， <italic>P</italic><0.05） of mice in each administration group was significantly shortened. The discrimination and exploration time of mice in the JY， WYJY， and FLU groups was significantly reduced （<italic>P</italic><0.01）， and the expression of Iba-1 in hippocampal CA1 region of mice in each administration group was significantly down-regulated （<italic>P</italic><0.01）. Conclusion:The warming Yang and relieving depression method helps to inhibit the occurrence and development of depression due to its efficacy in activating microglia in hippocampus of depression mice and lowering the expression of IL-1<italic>β</italic>， IL-6， and TNF-<italic>α</italic>.

NANOG Alleviates the Damage of Human Hair Follicle Mesenchymal Stem Cells Caused by H2O2 through Activation of AKT Pathway / 生物医学与环境科学（英文）

OBJECTIVE@#To explore the protective effect of NANOG against hydrogen peroxide (H2O2) -induced cell damage in the human hair follicle mesenchymal stem cells (hHF-MSCs).@*METHODS@#NANOG was expressed from a lentiviral vector, pLVX-IRES-ZsGreen. NANOG hHF-MSCs and vector hHF-MSCs were treated with 400 μmol/L hydrogen peroxide (H2O2) for 2 h, the cell survival rate, cell morphology, ROS production, apoptosis and expression of AKT, ERK, and p21 were determined and compared.@*RESULTS@#Our results showed that NANOG could activate AKT and upregulate the expression of p-AKT, but not p-ERK. When treated with 400 μmol/L H2O2, NANOG hHF-MSCs showed higher cell survival rate, lower ROS production and apoptosis, higher expression of p-AKT, higher ratio of p-AKT/AKT.@*CONCLUSION@#Our results suggest that NANOG could protect hHF-MSCs against cell damage caused by H2O2 through activating AKT signaling pathway.

Transcatheter Arterial Chemoembolization Combined with Simultaneous Computed Tomography-guided Radiofrequency Ablation for Large Hepatocellular Carcinomas / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Currently, the treatment of large hepatocellular carcinoma (HCC) is still a challenging problem. Transcatheter arterial chemoembolization (TACE) is the main treatment for intermediate end-stage HCC, while it is only a palliative and not a curative treatment due to the existence of residual tumors, and radiofrequency ablation (RFA) has limitations in complete ablation of large HCC. We hypothesized that TACE combined with simultaneous RFA (herein referred to as TACE + RFA) could improve the efficacy and survival of large HCC. This study aimed to investigate the feasibility, efficacy, and safety of TACE + RFA on single large HCC.</p><p><b>METHODS</b>A total of 66 patients with single large HCC (≥5 cm in diameter) were recruited between February 2010 and June 2016. TACE was first performed and computed tomography was performed immediately after TACE, and the lesions with poor lipiodol deposition were subjected to simultaneous RFA. The success rate, technique-related complications, liver and kidney functions, serum alpha-fetoprotein (AFP) levels, progression-free survival (PFS), median survival time (MST), focal control rate, and long-term survival rate were evaluated.</p><p><b>RESULTS</b>TACE + RFA were performed smoothly in all the patients with the success rate of 100%. Intra- and post-operative severe complications were not observed. There were no marked differences in mean alanine transaminase or aspartate transaminase before TACE + RFA compared with 7 days after TACE + RFA (all P > 0.05). In 57 AFP-positive patients, the levels of serum AFP were reduced by 100.0%, 100.0%, and 94.7% at 1, 3, and 6 months after TACE + RFA, respectively; the tumor control rates (complete remission + partial remission) were 100.0% (66/66), 92.4% (61/66), 87.9% (58/66), and 70.1% (39/55) at 1, 3, 6, and 12 months after TACE + RFA, respectively. Patients were followed up for 7-82 months after TACE + RFA. The MST was 18.3 months, PFS was 14.2 ± 6.2 months, and the 1-, 3-, and 5-year survival rates were 93.2% (55/59), 42.5% (17/40), and 27.2% (9/33), respectively.</p><p><b>CONCLUSION</b>TACE + RFA is safe, feasible, and effective in enhancing the focal control rate and survival rate of patients with large HCC.</p>

CT enhancement value for staging untypical T4a gastric cancer / 实用放射学杂志

Objective To evaluate the diagnostic performance of enhancement value and morphological features by using mul-tiphasic MDCT on differentiating untypical T4a from T3 gastric cancer.Methods Fifty-one histopathologically proven T3 and T4a gastric cancer patients with smooth serosa were collected retrospectively.Three radiologists read all images regarding morphological features,while the CT value and enhancement value of regions of interest (ROIs)located in the outer of lesion were calculated.Cut-off analysis was performed to determine optimal threshold levels of enhancement value to discriminate T4a and T3 gastric cancer.Di-agnostic performance of enhancement value and combination of enhancement value and morphologic assessment were compared with morphologic assessment by means of receiver operating characteristic (ROC)curve analysis.Results The sensitivity and specificity of morphological features was 66.67% and 33.33% respectively;the area under the ROC of enhancement value (between venous phase and plain scan)for differentiating T4a from T3 gastric cancer was 0.82,with a cut-off at 43.6 HU,sensitivity of 74.07% and specificity of 70.83%.Combined conventional standard and enhancement value,sensitivity of 100.00% and specificity 26.31%. Conclusion For differentiation of T4a and T3 gastric cancer by means of MDCT,enhancement value is found to be superior to con-ventional standard.