Yinlai Decoction Protects Microstructure of Colon and Regulates Serum Level of D-Lactic Acid in Pneumonia Mice Fed with High-Calorie and High-Protein Diet / 中国结合医学杂志

OBJECTIVE@#To investigate the effect of Yinlai Decoction (YD) on the microstructure of colon, and activity of D-lactic acid (DLA) and diamine oxidase (DAO) in serum of pneumonia mice model fed with high-calorie and high-protein diet (HCD).@*METHODS@#Sixty male Kunming mice were randomly divided into 6 groups by the random number table method: normal control, pneumonia, HCD, HCD with pneumonia (HCD-P), YD (229.2 mg/mL), and dexamethasone (15.63 mg/mL) groups, with 10 in each group. HCD mice were fed with 52% milk solution by gavage. Pneumonia mice was modeled with lipopolysaccharide inhalation and was fed by gavage with either the corresponding therapeutic drugs or saline water, twice daily, for 3 days. After hematoxylin-eosin staining, the changes in the colon structure were observed under light microscopy and transmission electron microscope, respectively. Enzyme-linked immunosorbent assay was used to detect the protein levels of DLA and DAO in the serum of mice.@*RESULTS@#The colonic mucosal structure and ultrastructure of mice in the normal control group were clear and intact. The colonic mucosal goblet cells in the pneumonia group tended to increase, and the size of the microvilli varied. In the HCD-P group, the mucosal goblet cells showed a marked increase in size with increased secretory activity. Loose mucosal epithelial connections were also observed, as shown by widened intercellular gaps with short sparse microvilli. These pathological changes of intestinal mucosa were significantly reduced in mouse models with YD treatment, while there was no significant improvement after dexamethasone treatment. The serum DLA level was significantly higher in the pneumonia, HCD, and HCD-P groups as compared with the normal control group (P<0.05). Serum DLA was significantly lower in the YD group than HCD-P group (P<0.05). Moreover, serum DLA level significantly increased in the dexamethasone group as compared with the YD group (P<0.01). There was no statistical significance in the serum level of DAO among groups (P>0.05).@*CONCLUSIONS@#YD can protect function of intestinal mucosa by improving the tissue morphology of intestinal mucosa and maintaining integrity of cell connections and microvilli structure, thereby reducing permeability of intestinal mucosa to regulate the serum levels of DLA in mice.

Changes in Intestinal Microflora and Lipid Metabolism in Ovariectomized Rats and Regulatory Effect of Erxian Decoction / 中国实验方剂学杂志

ObjectiveTo explore the intervention effect of Erxian decoction on intestinal microflora after ovariectomy in rats by 16S rRNA gene sequencing. MethodThirty-two female healthy SD rats were randomly divided into a Sham operation （Sham） group， a model （OVX） group， an estrogen （E） group， and an Erxian decoction （EXD） group， with 8 rats in each group. The rats in the E group and the EXD group received 1.8×10-4 g·kg-1 estradiol valerate solution and 9 g·kg-1 Erxian decoction， respectively， and those in the Sham group and the OVX group received an equal volume of distilled water once a day for 16 weeks. After 16 weeks， the levels of serum estrogen and blood lipid were detected. The fecal DNA was extracted， followed by 16S rRNA gene sequencing and analysis. ResultCompared with the Sham group， the OVX group showed reduced serum estrogen level （P<0.01） and increased serum levels of total cholesterol （TC） and low-density lipoprotein cholesterol （LDL-C） （P<0.05）. Compared with the OVX group， the E group and the EXD group showed increased serum estrogen level （P<0.01） and reduced TC and LDL-C （P<0.05）. Alpha diversity showed that there was no significant change in intestinal microflora diversity after ovariectomy. Beta diversity showed that there were significant differences in the structure of intestinal microflora in the four groups. The intervention of Erxian decoction could improve the changes in intestinal microflora after ovariectomy. LEfSe was used to analyze the differential flora in the four groups. The results showed that the Sham group and the OVX group had 3 differential bacterial phyla and 18 differential bacterial genera， the OVX group and the E group had 1 differential bacterial phylum and 12 differential bacterial genera， and the OVX group and the EXD group had 3 differential bacterial phyla and 5 differential bacterial genera. Estrogen intervention could reverse the change trend of Ruminococcus 1， Anaerovibrio， and Turicibacter in the OVX group. Erxian decoction intervention could reverse the change trend of Bacteroidetes， Firmicutes， Prevotella 9， Ruminococcaceae UCG-014， Ruminococcus 1， and Fusicatenibacter in the OVX group. ConclusionThe structure and function of intestinal microflora in ovariectomized rats changed obviously， and Erxian decoction could ameliorate the change.

Baihutang Syndrome and Treatment： A New Interpretation Based on Six Dimensional Syndrome Differentiation Theory / 中国实验方剂学杂志

Six dimensional syndrome differentiation theory， put forward by professor GU Xiao-hong at Beijing University of Chinese Medicine based on her theoretical teaching and clinical experience， emphasizes that the syndrome differentiation should be carried out from six dimensions including etiology， disease location， disease stage， disease condition， pathology， and disease nature， which is conducive to clinical thinking training and formation of traditional Chinese medicine （TCM）. The differentiation and treatment of Baihutang syndrome frequently seen in cold damage and warm disease still need to be explored. Guided by the six dimensional syndrome differentiation theory coupled with diverse viewpoints of cold damage and warm disease schools， this paper summarized and reinterpreted the understandings and thoughts of GU Xiao-hong and YU He， warm disease specialists of two generations. Considering the lung-stomach dysfunction caused by the internal invasion of exogenous pathogens， Baihutang syndrome was staged into Qi aspect. In this stage， exuberant pathogens and sufficient healthy Qi allowed the prevailing of internal heat and the consumption of body fluid， manifested as high fever， profuse sweating， thirst， and the pulse corresponding to interior excess and heat syndrome. This paper also pointed out that the Baihutang syndrome involved both lung and stomach， and the adoption of Baihutang contributed to preventing tu from restricting shui in the case of extreme excess of Yang brightness and protecting the kidney Yin. As revealed by the dynamic analysis of prognosis of Baihutang syndrome based on the six dimensional syndrome differentiation theory， even though the Baihutang syndrome could be present in both cold damage and warm disease， the specific disease stage， transmission and change， condition， prognosis， pathology， and medication differed. On this basis， a series of prescriptions have been modified from Baihutang， which has expanded the application scope of Baihutang and enriched its research value， thus better promoting its clinical application.

Proliferation, Differentiation and Immunoregulatory Capacities of Brown and White Adipose-Derived Stem Cells from Young and Aged Mice

Background and Objectives@#Adipose tissue is a source of mesenchymal stem cells, which have the potential to differentiate into various types of cells. Adipose-derived stem cells (ADSCs) are now recognized as an accessible, abundant, and reliable stem cells suitable for tissue engineering and regenerative medicine applications. However, few literatures gave a comprehensive report on the capacities of ADSCs harvested from different sites. Especially, the capacities of ADSCs from aged mice remained unclear. In this study, we investigated several main capacities of brown adipose derived stem cells (B-ADSCs) and white adipose derived stem cells (W-ADSCs) from both young and aged mice. @*Methods@#and Results: When isolated from young mice, B-ADSCs showed a stronger proliferation rate and higher osteogenic, adipogenic and myocardial differentiation ability than W-ADSCs. Carboxy fluorescein diacetate succinimidyl ester (CFSE) labeling test suggested no significant difference in immunosuppression capacity between B-ADSCs and W-ADSCs. Similarly, no difference between these two were found in several immune related molecules, such as programmed death-ligand 1 (PD-L1), intercellular cell adhesion molecule (ICAM-1), vascular cell adhesion molecule (VCAM-1), inducible nitric oxide synthase (iNOS), tumour necrosis factor-α (TNF-α), interleukin 10 (IL10), and suppressor of cytokine signaling 1 (socs1). When isolated from aged mice, B-ADSCs also showed a stronger proliferation rate and higher osteogenic, adipogenic and myocardial differentiation ability than W-ADSCs; however, it demonstrated an attenuated immunosuppression capacity compared to W-ADSCs. @*Conclusions@#In summary, our data showed that ADSCs’ characteristics were tissue source dependent and changed with age. It provided evidence for choosing the right tissue-specific ADSCs for clinical application and fundamental research.

Effect of Erxiantang on Cardiac Function and Blood Pressure Dynamic Evolution in Ovariectomized Rats / 中国实验方剂学杂志

Objective::To investigate the evolution of cardiac function and blood pressure in ovariectomized rats and the effect and mechanism of Erxiantang. Method::Healthy 10-week-old female SPF SD rats were randomly divided into sham operation group, model group, estrogen group(estradiol valerate, 0.18 mg·kg-1) and Erxiantang group(7.5 g·kg-1). The rats were intragastrically administered 2 weeks after ovariectomy, once a day for 12 weeks.Sham operation groups and model groups were given equal volumes of purified water.At the 4th week, 8th week, and 12th week after administration, the cardiac function, blood pressure, and levels of estrogen (E2) in rat serum were measured by non-invasive ultrasound cardiogram (UCG), tail artery detection techniques and radioimmunoassay.The levels of endothelin-1 (ET-1) and angiotensin 2(Ang Ⅱ) in rat serum were detected by enzyme-linked immuno sorbent assay (ELISA). The cardiac morphology and apoptosis were detected by hematoxylin-eosin (HE) staining, electron microscopy and Terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL). Result::Compared with sham operation group, the ejection fraction (EF) decreased and the left ventricular end systolic volume (LVVols) increased in the model group at 4th week after administration(P<0.05). There was no significant difference in cardiac function between the groups at 8th week.The left ventricular end diastolic diameter (LVIDs), LVVols, left ventricular end diastolic diameter (LVIDd), and left ventricular end diastolic volume (LVVold) were significantly increased in the model group at 12th week (P<0.01). At the 4th weeks, 8th week and 12th week, the systolic blood pressure (SBP) of the model group increased (P<0.05) and showed an increasing trend, and the diastolic blood pressure (DBP) did not change significantly.At the 12th week, the levels of E2 in serum decreased (P<0.05), ET-1 and Ang Ⅱ increased of the model group (P<0.01). The cardiac myofibrils were irregular, some myofilament was broken, and mitochondrial palsy was disordered, broken or disappeared, and cardiac apoptosis increased (P<0.01). Compared with the model group, myocardial contraction and diastolic function were significantly improved in Erxian decoction group, and blood pressure was decreased.The levels of E2 in serum was increased (P<0.05). The levels of ET-1 was decreased (P<0.05), and AngⅡ in serum was significantly decreased (P<0.01). The mitochondrial morphological structure was improved and the cardiac apoptotic rate was significantly decreased (P<0.01). Conclusion::After the ovariectomy, the rats showed a series of pathological changes such as decreased heart function and increased blood pressure.Compared with the decrease of heart function, the changes of blood pressure appeared earlier.Erxiantang exerts its intervention on cardiac function and blood pressure in ovariectomized rats by regulating E2, blood active substances and cardiac apoptosis.

Effect of Erxian Decoction on Myocardial Microvessels and Hemorheology in Ovariectomized Rats / 中国实验方剂学杂志

Objective:To observe the changes of myocardial microvessel density， microvascular endothelial cell morphology and hemorheology in ovariectomized rats and explore the interventional effects of Erxian decoction. Method:Thirty-two healthy 10 week-old female SPF SD rats were randomly divided into sham operation group， model group， estrogen group （estradiol valerate， 0.18 mg·kg-1） and Erxian decoction group （9 g·kg-1）. The rats were intragastrically administered 2 weeks after ovariectomy， once a day for 16 weeks. Sham operation groups and model groups were given equal volumes of purified water. After 16 weeks of administration， the cardiac function was measured by noninvasive ultrasound cardiogram （UCG）， CD34 in the myocardial tissue was tested by immunofluorescence staining to measure the microvessel density， the morphological structure of microvessels of myocardial tissue were detected by transmission electron microscope， the levels of estrogen （E2） in rat plasma were detected by radioimmunoassay， the levels of endothelin-1 （ET-1）， prostacyclin I2 （PGI2）， thromboxane A2 （TXA2）， endothelial growth factor （VEGF）， and von Willebrand Factor （vWF） in rat plasma were detected by enzyme-linked immuno sorbent assay （ELISA）， four items of coagulation was detected by blood coagulation analyzer， whole blood viscosity and plasma viscosity were detected by hemorheology. Result:Compared with sham operation group， the ejection fraction （EF） decreased （P<0.01）， the left ventricular short axis shortening rate （FS） decreased （P<0.01）， and the left ventricular end systolic volume （LVVols） increased （P<0.01）， myocardial microvessel density significantly reduced （P<0.01）， the endothelial cells were swollen and the cytoplasm was cavitation， E2 in rat plasma decreased （P<0.01）， ET-1， VEGF， vWF increased （P<0.01）， prostacyclin I2 /thromboxane A2 （PGI2/TXA2） decreased （P<0.01）， plasma activated partial prothrombin time （APTT） decreased （P<0.01）， fibrinogen （FIB） increased （P<0.01）， whole blood viscosity， plasma viscosity， and cassone viscosity increased （P<0.01）， whole blood high-cut， low-cut index， and red blood cell （RBC） aggregation index increased （P<0.05） in model group. Compared with model group， EF and FS increased （P<0.05）， LVVols decreased （P<0.05）， myocardial microvessel density significantly increased （P<0.01）， the endothelial cell edema was improved， and transport vesicles were clearly visible， E2 in rat plasma increased （P<0.01）， ET-1， VEGF， decreased （P<0.01）， PGI2/TXA2 increased （P<0.01）， APTT increased （P<0.01）， whole blood viscosity， whole blood high shear relative index， RBC aggregation index decreased （P<0.05）， Kasson viscosity and plasma viscosity decreased （P<0.01） in Erxian decoction group. Conclusion:Erxian decoction increases myocardial microvessel density， protects the structural integrity of microvascular endothelial cells， improves its endothelial secretion function and hemorheology in ovariectomized rats， and protects heart function.

Deubiquitinase MYSM1 Regulates Differentiation of Human B Cells to Plasma Cells / 中国实验血液学杂志

OBJECTIVE@#To study the regulatory effect of deubiquitinase MYSM1 on differentiation of B cells to plasma cells.@*METHODS@#The interfering and overexpression plasmids of MYSM1 were constructed and then the corresponding lentiviruses were packaged. Human CD19 B cells were isolated from human peripheral blood with Miltenyi B cell isolation kit. Purified CD19 B cells were transduced with lentiviruses and then treated with LPS, the CD138 expression was detected by flow cytometry. The expression of transcription factor was determined by quantitative PCR.@*RESULTS@#The differentiation of B cells to plasma cells was enhanced after interfering in MYSM1 expression. Quantitative PCR showed that mRNA levels of Pax5 and Bach2 in cells with interfering in MYSM1 were much lower than their counterpart (P＜0.01), and mRNA levels of Prdm1 and Xbp1 in cells with interfering in MYSM1 were much higher than their counterpart (P＜0.01). On the contrary, the differentiation of B cells to plasma cells was inhibited after the overexpression of MYSM1. Quantitative PCR showed that mRNA levels of Pax5 and Bach2 in cells with MYSM1 overexpression were higher than those in control cells (P＜0.01), and mRNA levels of Prdm1 and Xbp1 in cells with MYSM1 overexpression were much lower than those in their counterpart (P＜0.01).@*CONCLUSION@#MYSM1 negatively regulates differentiation of human B cells to plasma cells.

MicroRNA-20a Promotes Osteogenic Differentiation of C3H/10T1/2 Cells through Regulating CKIP-1 Expression / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of microRNA-20a(MiR-20a) on osteogenic differentiation of mouse C3H/10T1/2 cells and its regulatory mechanism.</p><p><b>METHODS</b>Osteogenic differentiation of C3H/10T1/2 cells were identified by ALP staining and qRT-PCR. MiR-20a mimics and CKIP-1 siRNA were transfected into C3H/10T1/2 cells respectively with lipo3000. The expression of osteoblast marker genes, miR-20a and CKIP-1 were quantitatively assessed by qRT-PCR.</p><p><b>RESULTS</b>miR-20a expression was up-regulated during osteoblast differentiation of C3H/10T1/2 cells. Overexpression of miR-20a promoted osteogenic differentiation. Furthermore, miR-20a inhibited the expression of bone formation negative regulator CKIP-1. Additionally, CKIP-1 knockdown promoted osteogenic differentiation.</p><p><b>CONCLUSION</b>MiR-20a promotes osteogenic differentiation of C3H/10T1/2 cells possibly through inhibiting the expression of CKIP-1.</p>

Regulating effects of whole-body vibration on protein expression of p-GSK3β in bone marrow cells of ovariectomized osteoporosis rats / 生理学报

The aim of this study was to investigate the effects of whole-body vibration on Wnt/β-catenin signaling pathway in bone marrow cells of ovariectomized osteoporosis rats. Thirty-six healthy 3-month old female Sprague Dawley (SD) rats were randomly divided into the following three groups by body weight: sham-operation (Sham), ovariectomized (OVX), and OVX whole-body vibration (WBV) groups. Ten weeks after ovariectomization, the rats of WBV group received vibration treatment (90 Hz, 15 min) twice per day. At the end of 8-week vibration, the whole-body bone mineral density (BMD) and body composition were detected by dual energy X-ray absorptiometry (DEXA) in vivo. The protein expressions of β-catenin and p-GSK3β in both bone marrow cells and bone marrow stromal cells were detected by Western blot. The results showed that, compared with OVX group, WBV group showed decreased fat mass and fat mass content, as well as increased lean body mass content. The BMD of the proximal tibia in WBV group was significantly higher than that in OVX group, however, there was no difference of BMD in whole-body and other positions between the two groups. The β-catenin expression in bone marrow stromal cells showed no difference between OVX and WBV groups. The p-GSK3β expression of bone marrow cells was increased in WBV group compared with that in OVX group, whereas bone marrow stromal cells from two groups did not exhibit the difference of the p-GSK3β expression. These results suggest that whole body vibration can stimulate the protein expression of p-GSK3β in bone marrow cells of ovariectomized osteoporosis rats, which could improve the bone loss induced by ovariectomization.