Identification of chemical components and network pharmacology of Huanglian Decoction based on UPLC-Q-TOF-MS/MS technology / 中国中药杂志

The chemical components of Huanglian Decoction were identified by ultra-performance liquid chromatography-quadrupole-time-of-flight-tandem mass spectrometry(UPLC-Q-TOF-MS/MS) technology. The gradient elution was conducted in Agilent ZORBAX Extend-C_(18) column(2.1 mm×100 mm, 1.8 μm) with the mobile phase of 0.1% formic acid aqueous solution(A)-acetonitrile(B) at a flow rate of 0.3 mL·min~(-1) and the column temperature of 35 ℃. The MS adopted the positive and negative ion mode of electrospray ionization(ESI), and the MS data were collected under the scanning range of m/z 100-1 500. Through high-resolution MS data analysis, combined with literature comparison and confirmation of reference substances, this paper identified 134 chemical components in Huanglian Decoction, including 12 alkaloids, 23 flavonoids, 22 terpenes and saponins, 12 phenols, 7 coumarins, 12 amino acids, 23 organic acids, and 23 other compounds, and the medicinal sources of the compounds were ascribed. Based on the previous studies, 7 components were selected as the index components. Combined with the network pharmacology research and analysis me-thods, the protein and protein interaction(PPI) network information of the intersection targets was obtained through the STRING 11.0 database, and 20 core targets of efficacy were screened out. In this study, UPLC-Q-TOF-MS/MS technology was successfully used to comprehensively analyze and identify the chemical components of Huanglian Decoction, and the core targets of its efficacy were discussed in combination with network pharmacology, which laid the foundation for clarifying the material basis and quality control of Huanglian Decoction.

Role of interaction between reactive oxygen species and ferroptosis pathway in methylglyoxal-induced injury in mouse embryonic osteoblasts / 南方医科大学学报

OBJECTIVE@#To explore the interaction between reactive oxygen species (ROS) and ferroptosis in methylglyoxalinduced injury of mouse embryonic osteoblasts (MC3T3-E1 cells).@*METHODS@#MC3T3-E1 cells were treated with methylglyoxal to establish a cell model of diabetic osteoporosis. CCK-8 assay was used to detect the viability of MC3T3-E1 cells. Rhodamine 123 staining followed by photofluorography was used to examine mitochondrial membrane potential (MMP). The intracellular ROS level was detected by 2', 7'-dichlorodihydrofluorescein diacetate staining with photofluorograph. Alkaline phosphatase (ALP) activity in the cells was detected using an ALP kit, the number of mineralized nodules was determined with alizarin red S staining, and the level of iron ions was detected using a detection kit. The expression level of glutathione peroxidase 4 (GPX4, a marker protein that inhibits ferroptosis) in the osteoblasts was determined using Western blotting.@*RESULTS@#Treatment of MC3T3-E1 cells with 0.6 mmol/L methylglyoxal for 24 h significantly inhibited the expression level of GPX4 (P < 0.001), increased intracellular iron ion concentration, decreased the cell viability, increased the loss of MMP and intracellular ROS level, decreased both ALP activity and the number of mineralized nodules in the cells (P < 0.001). Co-treatment of MC3T3-E1 cells with 2 mmol/L N-acetylcysteine (NAC, a ROS scavenger) and methylglyoxal significantly increased the expression level of GPX4 (P < 0.01); co-treatment with 4 mmo/L FER-1 (a ferroptosis inhibitor) and methylglyoxal obviously decreased the intracellular ROS level (P < 0.001). Co-treatment of the cells either with NAC and methylglyoxal or with FER-1 and methylglyoxal attenuated methylglyoxal-induced injuries in the osteoblasts (P < 0.001).@*CONCLUSION@#The interaction between ROS and ferroptosis pathway plays an important role in methylglyoxal-induced injury of mouse embryonic osteoblasts.

Discussion on the teaching design and practice of Clinical Medicine Summary / 中华医学教育探索杂志

Applying the effective teaching design to the teaching of Clinical Medicine Summary can improve the teaching quality of the course, the learning enthusiasm and effectiveness of non-clinical major students, so as to achieve the teaching purpose of the course. Practice has proved that optimizing the professional curriculum content, rationally using the information resources to mobilize students study initiative, improving teachers' own quality to improve teaching methods and means, changing the test standards and assessment methods and other effective teaching designs can significantly improve the teaching quality of the course, the students' learning motivation and learning effect, and achieve the teaching objectives.

Characteristics of neutrophil extracellular traps in patients with periodontitis and gingivitis

Abstract We sought to compare the characteristics and clinical significance of neutrophil extracellular traps in gingival samples from patients with periodontitis and those with gingivitis. The clinical indexes of gingival samples from patients with periodontitis and gingivitis were measured; the expression of TNF-alpha and IL-8 was measured by real-time fluorescent quantitative PCR; and the expression of TLR-8 and MMP-9 was measured by western blotting assays. Chemotaxis, phagocytosis and phagocytic activity of neutrophils were measured. Compared with the healthy group, the expression of TNF-α and IL-8 in the periodontitis group and the gingivitis group increased significantly (p < 0.05), and TNF-α in the gingivitis group was significantly lower than that in the healthy group (p < 0.05). The expression of IL-8 in the periodontitis group was significantly higher than that in the periodontitis group (p < 0.05). Furthermore, the expression of TLR-8 and MMP-9 in the periodontitis group was different from that in the gingivitis group and the healthy group, and the expression of TLR-8 and MMP-9 in the gingivitis group was significantly different from that in the healthy group (p < 0.05). In addition, the neutrophil mobility index in healthy people was 3.02 ± 0.53, that in the periodontitis group was 2.21 ± 0.13, and that in the gingivitis group was 2.31 ± 0.12. In conclusion, the chemotaxis of neutrophils in gingival samples of patients with periodontitis and gingivitis was decreased, the phagocytotic ability and activity of neutrophils were reduced, and the release of the extracellular trap-releasing inducible factors TNF-alpha and IL-8 also declined.

Application of fingerprint combined with QAMS in quality evaluation of Aurantii Immaturus Fructus / 中草药

Objective: This paper aims to provide the experimental foundation for quality control of Aurantii Immaturus Fructus from different habitats by analyzing the multi-component qualitatively and quantitatively. Methods: Fingerprint pattern and QAMS were combined in this experiment, and HPLC analysis was performed on a Waters X-Brige C18 (200 mm × 4.6 mm, 5 μm) column with mobile phase composed of acetonitrile (A) -0.1% methanoic acid solution at a flow rate of 1.0 mL/min in gradient elution mode. The column temperature was maintained at 30 ℃, the injection volume was 10 μL, and the detection wavelengths were set at 283 and 330 nm. Results: The HPLC-UV fingerprint of Aurantii Immaturus Fructus from different habitats was established with 21 common peaks, 9 of them were identified as narirutin, naringin, hesperidin, neohesperidin, naringenin, hesperetin, sinensetin, nobiletin and tangeretin, and the similarity of 23 batches of samples were greater than 0.9, CA and PCA show the classification is consistent with the habitats distribution of the medicine. Then, the QAMS result showed some differences with neohesperidin to be the internal standard. Conclusion: From the perspective of composition, there are some differences in the quality of Aurantii Immaturus Fructus from different habitats, and the method of QAMS is accurate, efficient and feasible, which has obvious advantages than standard curve method and external standard method. In a word, it can be applied to quality evaluation and provides the experimental foundation for quality control of Aurantii Immaturus Fructus from different habitats.

Investigation of aflatoxins and toxigenic fungi contamination during post-harvest processing of Polygalae Radix / 中草药

Objective: To investigate aflatoxins contamination B1, B2, G1, G2 (AFB1, ATB2, AFG1, AFG2), toxigenic fungi species and potential contamination sources of Polygalae Radix during post-harvest processing, and analyze the main ways of aflatoxins contamination. Methods: Twenty-one Polygalae Radix samples were collected from multiple steps during the post-harvest processing in this study. Aflatoxin levels in these samples were determined by immunoaffinity column and HPLC coupled with post-column photochemical derivatization. Dilution-plate method was applied for the fungi isolation followed by strain identification based on morphological characterization and molecular approaches. Results: Aflatoxins were detected in 15 samples, but none of them exceeded the limit set by Chinese Pharmacopoeia. The fungal counts increased significantly from newly harvested samples to post-sweating, and the counts further increased to the maximum (2 × 108 CFU/g) after xylem-removing, then decreased after drying. In contrast, fungal counts of samples dried directly after harvesting did not change much throughout the processing. There was a significant positive correlation between fungal counts and water activity (Aw). A total of 209 fungal belonged to five genera were identified from the samples, and Penicillium was the predominant genus. Cladosporium and Fusarium were increased after sweating, and then Aspergillus increased after xylem-removing and drying. One A. parasiticus strain was confirmed to be able to produce AFB1, AFB2, AFG1, and AFG2. Conclusion: Aflatoxins contamination happened in both field production and post-harvest processing of Polygalae Radix. Especially, the contamination of Penicillium spp. should be paid more attention.

Q-marker prediction analysis of Paeoniae Radix Alba based on fingerprint and network pharmacology / 中草药

Objective: To explore the potential Q-marker of Paeoniae Radix Alba in Danggui Sini Decoction based on fingerprint and network pharmacology. Methods: The fingerprints of Paeoniae Radix Alba decoction and Danggui Sini Decoction were established, and the law of components transfer was also defined. The "compounds-targets-pathways" network was then established to predict the potential Q-marker of Paeoniae Radix Alba through the network pharmacology. Results: The fingerprints of 15 batches of Paeoniae Radix Alba decoction and 15 batches of Danggui Sini Decoction were established, and the five chromatographic peaks were identified, they were gallic acid, albiflorin, paeoniflorin, 1,2,3,4,6-O-pentagalloylglucose, and benzoyl paeoniflorin. Through the network pharmacology analysis, the potential two active components, eight core targets and 13 key pathways were screened out, which indicated that paeoniflorin and albiflorin were preliminarily predicted to the potential Q-marker of Paeoniae Radix Alba. Conclusion: The analysis and prediction of the Q-marker in this study can provide a reference for the whole control of the Paeoniae Radix Alba quality, which can also provide the basis for the further research on the efficacy-related substance and mechanism of Paeoniae Radix Alba.

Investigation of aflatoxins,mycobiota,and toxigenic fungi during post-harvest handling of Ziziphi Spinosae Semen / 中国中药杂志

Ziziphi Spinosae Semen is one of the Chinese herbal medicine being susceptible to aflatoxins contamination. To investigate the sources of aflatoxins contamination and toxigenic fungi species on Ziziphi Spinosae Semen,32 samples were collected from multiple steps during the post-harvest processing in this study. Aflatoxins in these samples were determined by immunoaffinity column and HPLC coupled with post-column photochemical derivatization. The dilution-plate method was applied to the fungi isolation. The isolated fungi strains were identified by morphological characterization and molecular approaches. The results showed that aflatoxins were detected in 28 samples from every step during the processing of Ziziphi Spinosae Semen. Three samples were detected with aflatoxin B_1 and 2 samples with both aflatoxin B_1 and total aflatoxin exceeding the limit of Chinese Pharmacopoeia. Especially the samples from the washing step,with the highest detected amounts of AFB_1 and AFs were reached 94. 79,121. 43 μg·kg~(-1),respectively. All 32 samples were contaminated by fungi. The fungal counts on the newly harvested samples were 2. 20 × 10~2 CFU·g~(-1). Moreover,it increased as tphreocessing progresses,and achieved 1. 16×10~6 CFU·g~(-1) after washing. A total of 321 isolates were identified to 17 genera. Aspergillus flavus was the main source of aflatoxins during the processing and storage of Ziziphi Spinosae Semen. One isolate of A. flavus was confirmed producing AFB_1 and AFB_2. The fungal count was significantly increased by composting,and Aspergillus was the predominant genus after shell breaking. The contamination level of aflatoxins was increased by composting and washing.

Material Basis and Action Mechanism of Wendantang in Treatment of Nervous System Diseases / 中国实验方剂学杂志

Wendantang (WDT) is a classical traditional Chinese medicine prescription composed of Pinelliae Rhizoma, Bambuseae Caulis in Taenias, Aurantii Fructus Immaturus, Citri Reticulatae Pericarpium, Zingiberis Rhizoma Recens, Glycyrrhizae Radix et Rhizoma, with the effect in regulating Qi-movement and phlegm and relieving stomach and gallbladder. The clinical studies have proved that WDT has significant therapeutic effects on depression, insomnia, schizophrenia, Alzheimer's disease and other nervous system diseases, but wihtout systematic understanding of material basis and compatibility principle because of the complex chemical composition and the scattered research results. Focusing on the neurological diseases and based on the origin of ancient recipes and modern research examples, the author sorted out and summarized the active ingredients constituting the recipe, paid attention to the effect of the compatibility on the composition and efficacy transmission, and judged the rationality of composition intention and selection. On this basis, it comprehensively identifies the potential components and effective paths that can well treat the nervous system diseases, and had the overall understanding about mutual relationship between composition and efficiency. In this article, we expect to find its scientific basis of effective materials and the key technology of quality standards, and define the direction of future research, so as to provide valuable reference for secondary development and new preparations designed of classic prescriptions.

Septation of the Intrapericardial Arterial Trunks in the Early Human Embryonic Heart / 中华医学杂志(英文版)

<p><b>Background</b>Outflow tract (OFT) septation defects are a common cause of congenital heart disease. Numerous studies have focused on the septation mechanism of the OFT, but have reported inconsistent conclusions. This study, therefore, aimed to investigate the septation of the aortic sac and the OFT in the early embryonic human heart.</p><p><b>Methods</b>Serial sections of 27 human embryonic hearts from Carnegie stage (CS) 10 to CS19 were immunohistochemically stained with antibodies against α-smooth muscle actin (α-SMA) and myosin heavy chain.</p><p><b>Results</b>At CS10-CS11, the OFT wall was an exclusively myocardial structure that was continuous with the aortic sac at the margin of the pericardial cavity. From CS13 onward, the OFT was divided into nonmyocardial and myocardial portions. The cushion formed gradually, and its distal border with the OFT myocardium was consistently maintained. The aortic sac between the fourth and sixth aortic arch arteries was degenerated. At CS16, the α-SMA-positive aortopulmonary septum formed and fused with the two OFT cushions, thus septating the nonmyocardial portion of the OFT into two arteries. At this stage, the cushions were not fused. At CS19, the bilateral cushions were fused to septate the myocardial portion of the OFT.</p><p><b>Conclusions</b>Data suggest that the OFT cushion is formed before the aortopulmonary septum is formed. Thus, the OFT cushion is not derived from the aortopulmonary septum. In addition, the nonmyocardial part of the OFT is septated into the aorta and pulmonary trunk by the aortopulmonary septum, while the main part of the cushion fuses and septates the myocardial portion of the OFT.</p>

Clinical value of pulse indicator continuous cardiac output monitoring in treating acute kidney injury due to sepsis / 新乡医学院学报

Objective To evaluate the clinical value of pulse indicator continuous cardiac output monitoring in treating acute kidney injury(AKI) due to sepsis.Methods Sixty-two patients with AKI due to sepsis in the Central Hospital of Zhumadian from August 2013 to August 2016 were rolled in and divided into control group (34 cases) and observation group (28 cases) according to whether adopted pulse indicator continuous cardiac output monitoring.Six hours and 24 hours after fluid resuscitatinn,resuscitation fluid volume,heart rate(HR),central venous pressure(CVP),mean arterial pressure(MAP),vasoactive drugs dose of the two groups were observed.Treatment times,daily filtration volume of continuous renal replacement therapy and independent urine volume,serum creatinine level,the survival rate after treating for seven days in the two groups were compared.Results There was no significant difference in the HR,central venous pressure,mean arterial pressure at six hours after fluid resuscitation between the two groups(P ＜ 0.05).At 24 hours after fluid resuscitation,there was no siguificant difference in the HR in the control group compared with that at six hours after fluid resuscitation (P ＜ 0.05),the CVP and MAP were higher than those at six hours after fluid resuscitation(P ＜ 0.05);the HR in the observation group was lower than that at six hours after fluid resuscitation (P ＜ 0.05),MAP was higher than that at six hours after fluid resuscitation (P ＜0.05),but there was no significant difference in the CVP in the control group compared with that at six hours after fluid resuscitation (P ＜ 0.05).The HR and CVP at 24 hours after fluid resuscitation in the observation group were lower than those in the control group(P ＜0.05),while there was no no significant difference in the MAP between the two groups(P ＜ 0.05).There was no significant difference in the fluid resuscitation volume,the dose of noradrenaline and dobutamine after treating for six hours between the two groups (P ＜ 0.05).Twenty four hours after fluid resuscitation resuscitation fluid volume and dobutamine dose in the observation group were significantly lower than those in the control group (P ＜ 0.05).The duration of renal replacement therapy,the daily filtration volume and the blood creatinine after treating for seven days in the observation group were significantly lower than those in the control group (P ＜ 0.05),but the independent urine volume after treating for seven days in the observation group was significantly higher than that in the control group (P ＜ 0.05).After treating for seven days,there were 28 cases survival,the survival rate was 82.4％ (28/34),while there were 25 cases survival,the survival rate was 89.3％ (25/28);the difference of survival rate between the two groups was not statistically significant (x2 =0.59,P ＜ 0.05).Conclusion In patients with AKI due to sepsis,pulse indicator continuous cardiac output monitoring can be used to optimize fluid management,and it can improved the renal function.

Clinicopathologic study of pimary paget disease in esophagus / 临床与实验病理学杂志

Purpose To investigate the clinicopathological and diagnostic characteristics of primary Paget disease (PD ) in esophagus. Methods The clinical presentation, histological observation and immunohistochemical staining were analyzed in four cases of primary PD involved esophagus and related literatures were reviewed. Results The patients were all male, aged from 61 to 74 years old. All the tumors were originated from the mucosa of the esophagus. Histologically, the Paget cells showed a single or small nesting and acinar distribution in the esophage-al mucosa. Adenocarcinoma in situ were seen in 2 cases and squamous cell carcinoma was seen in one of them. Immunohisto-chemically, the Paget cells were typically strongly positive for Ckpan, CKL, and CK7, while negative for CKH, CK5/6, CK14, p63. Conclusion Primary esophagus PD is rare. It can develop alone in esophagus or accompanied with adenocarcinoma in situ, invasive squamous cell carcinoma. The correct diagnosis need detailed pathological observation, immunohistochemical ev-idence and medical history.

Intestinal Microbial Community Differs between Acute Pancreatitis Patients and Healthy Volunteers / 生物医学与环境科学（英文）

A case control study including 45 acute pancreatitis and 44 healthy volunteers was performed to investigate the association between intestinal microbial community and acute pancreatitis. High-throughput 16S rRNA gene amplicon sequencing was used to profile the microbiological composition of the samples. In total, 27 microbial phyla were detected and the samples of pancreatitis patients contained fewer phyla. Samples from acute pancreatitis patients contained more Bacteroidetes and Proteobacteria and fewer Firmicutes and Actinobacteria than those from healthy volunteers. PCoA analyses distinguished the fecal microbial communities of acute pancreatitis patients from those of healthy volunteers. The intestinal microbes of acute pancreatitis patients are different from those of healthy volunteers. Modulation of the intestinal microbiome may serve as an alternative strategy for treating acute pancreatitis.

Angiotensin-(1-7) protects cardiac myocytes against high glucose-induced injury by inhibiting ClC-3 chloride channels / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore whether angiotensin-(1-7) [Ang-(1-7)] protects cardiac myocytes against high glucose (HG)-induced injury by inhibiting ClC-3 chloride channels.</p><p><b>METHOD</b>H9c2 cardiac cells were exposed to 35 mmol/L glucose for 24 h to establish a cell injury model. The cells were treated with Ang-(1-7) or the inhibitor of chloride channel (NPPB) in the presence of HG for 24 h to observe the changes in HG-induced cell injury. Cell counter kit 8 (CCK-8) assay was used to test the cell viability, and the morphological changes of the apoptotic cells were detected using Hoechst 33258 staining and fluorescent microscopy. The intracellular level of reactive oxygen species (ROS) was examined by DCFH-DA staining, SOD activity in the culture medium was measured using commercial kits, and the mitochondrial membrane potential (MMP) of the cells was tested with rodamine 123 staining. The expression level of cardiac ClC-3 chloride channels was detected with Western blotting.</p><p><b>RESULTS</b>Exposure of H9c2 cardiac cells to 35 mmol/L glucose for 24 h markedly enhanced the expressions of cardiac ClC-3 channel protein (P<0.01). Co-treatment of the cells with 1 µmol/L Ang-(1-7) and HG for 24 h significantly attenuated HG- induced upregulation of ClC-3 channel protein expression (P<0.01). Co-treatment of the cells exposed to HG with 1 µmol/L Ang-(1-7) or 100 µmol/L NPPB for 24 h obviously ameliorated HG-induced injuries as shown by increased cell viability, enhanced SOD activity, decreased number of apoptotic cells, and reduced intracellular ROS generation and loss of MMP (P<0.01).</p><p><b>CONCLUSION</b>ClC-3 channels are involved in HG-induced injury in cardiac cells. Ang-(1-7) protects cardiac cells against HG-induced injury by inhibiting ClC-3 channels.</p>

Isolation and Chemotaxonomic Significance of Chemical Constituents from Rubus parvifolius / 中草药·英文版

Objective: To study the chemical constituents from the roots of Rubus parvifolius. Methods: The chemical constituents were extracted and purified by silica gel column chromatography. NMR spectra were used for structural identification. Results: Phytochemical study on the roots of R. parvifolius led to the isolation of one ceramide (1), two anthraquinones ( 2 and 3), four triterpenoids (4-7), two flavonoids (8 and 9), one fatty acid ester (10), and two sterols (11 and 12). Conclusion: Compound 1 is isolated from the plants of family Rosaceae for the first time, and compounds 2-5 are isolated from genus Rubus for the first time. Though R. parvifolius shares the same major chemical types (triterpenoid, flavonoid, and anthraquinone) with those of R. alceaefolius, a substituent of R. parvifolius, their individual constituents are different. In addition, R. parvifolius contains ceramide (1) with high concentration, while caffeoylquinic acid reported in R. alceaefolius has not been found in R. parvifolius. Furthermore, the results from our phytochemical study are consistent with the DNA phylogenic relationship between R. parvifolius and R. alceaefolius (two separated subgenera), suggesting that the substitution of the former species with the latter one in folk medicine might not be suitable.

Soy Protein Supplementation Reduces Clinical Indices in Type 2 Diabetes and Metabolic Syndrome

PURPOSE: Clinical trials have studied the use of soy protein for treating type 2 diabetes (T2D) and metabolic syndrome (MS). The purpose of this study was to outline evidence on the effects of soy protein supplementation on clinical indices in T2D and MS subjects by performing a meta-analysis of randomized controlled trials (RCTs). MATERIALS AND METHODS: We searched PubMed, EMBASE, and Cochrane databases up to March 2015 for RCTs. Pooled estimates and 95% confidence intervals (CIs) were calculated by the fixed-and-random-effects model. A total of eleven studies with eleven clinical variables met the inclusion criteria. RESULTS: The meta-analysis showed that fasting plasma glucose (FPG) [weighted mean difference (WMD), -0.207; 95% CI, -0.374 to -0.040; p=0.015], fasting serum insulin (FSI) (WMD, -0.292; 95% CI, -0.496 to -0.088; p=0.005), homeostasis model of assessment for insulin resistance index (HOMA-IR) (WMD, -0.346; 95% CI, -0.570 to -0.123; p=0.002), diastolic blood pressure (DBP) (WMD, -0.230; 95% CI, -0.441 to -0.019; p=0.033), low-density lipoprotein cholesterol (LDL-C) (WMD, -0.304; 95% CI, -0.461 to -0.148; p=0.000), total cholesterol (TC) (WMD, -0.386; 95% CI, -0.548 to -0.225; p=0.000), and C-reactive protein (CRP) (WMD, -0.510; 95% CI, -0.722 to -0.299; p=0.000) are significant reduced with soy protein supplementation, compared with a placebo control group, in T2D and MS patients. Furthermore, soy protein supplementation for longer duration (≥6 mo) significantly reduced FPG, LDL-C, and CRP, while that for a shorter duration (<6 mo) significantly reduced FSI and HOMA-IR. CONCLUSION: Soy protein supplementation could be beneficial for FPG, FSI, HOMA-IR, DBP, LDL-C, TC, and CRP control in plasma.

Antioxidant effect of salvianolic acid B on hippocampal CA1 neurons in mice with cerebral ischemia and reperfusion injury / 中国结合医学杂志

<p><b>OBJETIVE</b>To investigate the neuroprotective effects and underlying mechanisms of salvianolic acid B (Sal B) extracted from Salvia miltiorrhiza on hippocampal CA1 neurons in mice with cerebral ischemia reperfusion injury.</p><p><b>METHODS</b>Forty male National Institute of Health (NIH) mice were randomly divided into 4 groups with 10 animals each, including the sham group, the model group, the SalB group (SalB 22.5 mg/kg) and the nimodipine (Nim) group (Nim 1 mg/kg). A mouse model of cerebral ischemia and reperfusion injury was established by bilateral carotid artery occlusion for 30 min followed by 24-h reperfusion. The malondialdehyde (MDA) content, the nitric oxide synthase (NOS) activity, the superoxide dismutase (SOD) activity and total antioxidant capability (T-AOC) of the pallium were determined by biochemistry methods. The morphologic changes and Bcl-2 and Bax protein expression in hippocampal CA1 neurons were observed by using hematoxylineosin staining and immunohistochemistry staining, respectively.</p><p><b>RESULTS</b>In the SalB group, the MDA content and the NOS activity of the pallium in cerebral ischemia-reperfusion mice significantly decreased and the SOD activity and the T-AOC significantly increased, as compared with the model group (P<0.05 or P<0.01). The SalB treatment also rescued neuronal loss (P<0.01) in the hippocampal CA1 region, strongly promoted Bcl-2 protein expression (P<0.01) and inhibited Bax protein expression (P<0.05).</p><p><b>CONCLUSIONS</b>SalB increases the level of antioxidant substances and decreases free radicals production. Moreover, it also improves Bcl-2 expression and reduces Bax expression. SalB may exert the neuroprotective effect through mitochondria-dependent pathway on hippocampal CA1 neurons in mice with cerebral ischemia and reperfusion injury and suggested that SalB represents a promising candidate for the prevention and treatment of ischemic cerebrovascular disease.</p>

Effect of different irradiation doses on the establishment of murine cGVHD model after MHC matched spleen stem cell transplantation / 中国实验血液学杂志

This study was aimed to investigate the effect of different irradiation doses on the establishment of murine cGVHD model after MHC matched spleen stem cell transplantation. The male mouse BALB/c(H)-2d was totally irradiated with different radiation dose of (60)Co (TBI), then was infused with the same number of splenocytes from MHC matched DBA/2 male mice. After transplantation, the bodyweight, general appearance, hair changes, survival time and pathological damage were observed. The results indicated that compared to the control group (0 Gy) and the 7.0 Gy group, the mice irradiated with 7.5 Gy and 8.0 Gy showed cGVHD symptoms and obvious pathological damage. At the end of experiments (60 d after transplantation), all mice irradiated by 7.5 Gy survived while only 60% animals survived in the 8.0 Gy group. It is concluded that under infusion of 10(8) MHC matched splenocytes per mouse, 7.5 Gy irradiation is appropriate to efficiently establish cGVHD model. This study laid an important foundation for further studying the pathogenesis, biological characteristics, and intervention factors of cGVHD.

Effect of intercellular adhesion molecule-1 on the migration in vitro of murine mesenchymal stem cells and its related mechanism / 中国实验血液学杂志

This study was aimed to investigate the effect of intercellular adhesion molecule-1 (ICAM-1) on the migration in vitro of the murine mesenchymal stem cells (MSC) and its related mechanisms. The migration ability of murine MSC (C3H10T1/2), ICAM-1 transfected MSC (C3H10T1/2-MIGR1-ICAM-1) and empty vector-transfected MSC (C3H10T 1/2-MIGR1) were assayed in vitro by using the transwell system. Briefly, the cells were seeded on the membrane with 8 µm aperture and the fetal bovine serum was used as the chemotactic agent to induce MSC migration. The transmigrated cells were stained by crystal purple as well as DAPI for 8 h and 12 h respectively. The absolute cell numbers were counted and the migration rates of MSC were evaluated in each group. To explore the potential mechanisms which control the migration of MSC, the specific chemical inhibitors of MAPK pathway (SB203580, PD98059 and JNK inhibitor II) were added to the transwell system and the alteration of the MSC migration ability were evaluated at 12 h. The results showed that the migration ability at 8 h and 12 h of the ICAM-1-transfected MSC increased. Both absolute cell number and migration rate of MSC were significantly up-regulated by ICAM-1. Furthermore, the promoting effect of ICAM-1 on migration was partially suppressed by the inhibition of JNK/SAPK pathway. The transmigrated cell numbers and the migration rate decreased with the addition of JNK inhibitor II. However, the ICAM-1 promoting migration of MSC was not suppressed by the inhibitors for ERK/MAPK and p38/MAPK pathway did not work in the present study. It is concluded that ICAM-1 can induce mouse MSC migration in vitro, and the promoting effect is partially dependent on the activation of JNK/SAPK pathway.

Latest research progress on pathogenesis of chronic graft versus host disease and its related problems / 中国实验血液学杂志

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an effective method for the treating of malignant diseases of hematopoietic system or non-malignant proliferative diseases, but the occurrence of graft-versus-host disease (GVHD) limits the success rate of hematopoietic stem cell transplantation. Moreover, chronic graft-versus-host disease (cGVHD) is the main factor affecting the long-term survival rate and life quality of recipient after hematopoietic stem cell transplantation. In this article, the latest research progress of the pathogenesis of cGVHD and related problems are reviewed from the thymus, cytokines, T lymphocyte subsets, B lymphocytes and its secreted antibody.