Summary and Review of Animal Models for Chronic Atrophic Gastritis and Precancerous Lesions of Gastric Cancer / 中国实验方剂学杂志

Gastric cancer is a disease with high morbidity and mortality in the world, and also obtains attention in the global medicine. The occurrence of gastric cancer is a multi-stage and multi-factor process. A large number of epidemiological, pathological and clinical evidences have confirmed that the risk of gastric cancer in patients with chronic atrophic gastritis (CAG) is significantly correlated with the mortality of gastric cancer. Gastric mucosal atrophy, intestinal metaplasia (especially incomplete colonic metaplasia) and dysplasia are the main stages of precancerous lesions of precancerous lesions of gastric cancer (PLGC). Monitoring the condition of CAG patients, especially those with intestinal metaplasia and dysplasia, is of great significance for the discovery of early gastric cancer. CAG and PLGC are great significance in the pathological stage of gastric carcinogenesis. In recent years, more and more in-depth clinical and experimental studies have been carried out in this direction. So far, animal experiment is the main research way for CAG and PLGC disease, so it is very important to explore the modeling method. Choosing a stable and reliable model is the primary factor to study animal experiment. In view of the relationship between two diseases, this paper will summarize the methods of establishing animal models for CAG and PLGC in recent years, generally including chemical drug mutagenesis, physical stimulation, immune modeling, Helicobacter pylori infection replication and surgical modeling. Examples would be given for the application of various methods in the previous experiments, and the author would make a brief comment on the merits and demerits of these methods, which have been explored and successfully made by the author. This study would provide certain reference for the establishment and application of animal models in further CAG and PLGC experiments.

Stroke, platelet activating factor and receptor antagonists / 中国中药杂志

Stroke is an acute cerebrovascular disease with high morbidity, disability and mortality. The prevention and treatment conditions for stroke is severe all over the world. Antiplatelet aggregation is an effective treatment. Platelet activation factor (PAF) is another important medium in mediating platelet aggregation, which plays an important role in the pathogenesis of stroke. In recent years, PAF receptor antagonists have attracted international attention in the field of stroke prevention and treatment. In this review, we would summarize the classification, mechanism and drug characteristics of PAF receptor antagonists in order to provide the valuable guidance and direction for clinical medicine and research.

Xiayuxue Decoction (symbols; see text) attenuates hepatic stellate cell activation and sinusoidal endothelium defenestration in CCl4-induced fibrotic liver of mice / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of ancient Chinese medical formula Xiayuxue Decoction ([symbols; see text], XYXD) on activation of hepatic stellate cells (HSCs) and defenestration of sinusoidal endothelial cells (SECs) in CCl4-induced fibrotic liver of mice.</p><p><b>METHODS</b>High performance liquid chromatography was used to identify the main components of XYXD and control the quality of extraction. C57BL/6 mice were induced liver fibrosis by CCl4 exposure and administered with XYXD for 6 weeks simultaneously. Liver tissue was investigated by hematoxylin-eosin and Sirius-red staining. Sinusoidal fenestrations were observed by scanning electronic microscopy and fluorescent immunohistochemistry of PECAM-1 (CD31). Whole liver lysates were detected of α-smooth muscle actin (α-SMA) and type-I collagen by Western blot. Primary rat HSCs-T6 cells were analyzed by detecting α-SMA, F-actin, DNA fragmentation through confocal microscopy, Western blot, terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay and cellomics arrayscan, respectively.</p><p><b>RESULTS</b>Amygdalin and emodin in XYXD were identified. XYXD (993 mg/kg) inhibited Sirius red positive area up to 70.1% (P<0.01), as well as protein levels of α-SMA and type-I collagen by 42.0% and 18.5% (P<0.05) respectively. In vitro, XYXD (12.5 μg/mL, 50 μg/mL) suppressed the activation of HSCs and reversed the myofibroblastic HSCs into quiescent, demonstrated as inhibition of fluorescent F-actin by 32.3% and 46.6% (P<0.05). Besides, XYXD induced the apoptosis of HSC-T6 cells by 20.0% (P<0.05) and 49.5% (P<0.01), evidenced by enhanced TUNEL positivity. Moreover, ultrastructural observation suggested XYXD inhibited defenestration of SECs, which was confirmed by 31.1% reduction of protein level of CD31 (P<0.05).</p><p><b>CONCLUSIONS</b>XYXD inhibited both HSCs activation and SECs defenestration which accompany chronic liver injuries. These data may help to understand the underlying mechanisms of XYXD for prevetion of chronic liver diseases.</p>

Simultaneous determination of five nucleosides and nucleobases of Rehmannia glutinosa Libosch. by high performance liquid chromatography / 药学学报

This study is to establish a method for simultaneously determination of five nucleosides and nucleobases, including hypoxanthine, uridine, adenine, guanosine and adenosine in Rehmannia glutinosa Libosch. which was collected from different regions in China. A Diamonsil C18 column (250 mm x 4.6 mm, 5 microm) was used. Acetonitrile and 0.04 mol L(-1) potassium dihydrogen phosphate solution were adopted as mobile phase with gradient elution. The flow rate was 1 mL min(-1) and column temperature was 30 degrees C. The detection wavelength was at 254 nm. The method had good linearity over the range of 1.0 - 16.0 microg mL(-1) (r2 = 0.999 8), 5.0 - 80.0 microg mL(-1) (r2 = 0.999 8), 1.0 - 16.0 microg mL(-1) (r2 = 0.999 5), 1.25 - 20.0 microg mL(-1) (r2 = 0.999 8) and 1.0 - 16.0 microg mL(-1) (r2 = 0.999 8) for hypoxanthine, uridine, adenine, guanosine and adenosine, respectively. The average recoveries were between 98.8% and 100.7%. The content of hypoxanthine, uridine, adenine, guanosine and adenosine in Rehmannia glutinosa Libosch. from different regions was significantly different. This established method was sensitive and reliable for the quantification of five chemical constituents in Rehmannia glutinosa Libosch.

Purification and functional identification of the recombinant human CREG/myc-His glycoprotein / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To purify the recombinant human cellular repressor of EIA stimulated gene (hCREG)/myc-His glycoprotein and confirm the biological function of hCREG/myc-His which could inhibit the proliferation of human internal thoracic artery smooth muscle cells (HITASY) cultured in vitro.</p><p><b>METHODS</b>The recombinant hCREG/myc-His protein was purified with Ni-NTA column according to 6 x His affinity chromatographic theory. The recombinant hCREG/myc-His protein was desalted by HiTrap Desalting Column. The effect of recombinant hCREG/myc-His glycoprotein of different concentration (0.5 microg/ml, 1 microg/ml and 2 microg/ml) on proliferation of HITASY cells was studied by flow cytometric analysis and the effect of recombinant protein on proliferation of HITASY cells was confirmed by BrdU incorporation method.</p><p><b>RESULTS</b>The recombinant hCREG protein was purified with Ni-NTA column according to 6 x His affinity chromatographic theory. The concentration of recombinant hCREG protein which has been concentrated and desalted was determined to be 1.6 mg/ml and the purity of recombinant protein reached 92%. The protein was identified to be glycosylated. The recombinant hCREG protein was identified to inhibit the proliferation of HITASY cells cultured in vitro and the inhibition effect was stronger in low-dosage group than that in high-dosage group by flow cytometric analysis. The proliferation of HITASY cells cultured in vitro with 2 microg/ml recombinant hCREG protein was inhibited significantly compared with that in control group according to the BrdU incorporation result. There was statistical difference among the groups (P < 0.05).</p><p><b>CONCLUSION</b>The purification of recombinant hCREG/myc-His glycoprotein with biological activity provides an experiment platform for function study and engineering production of hCREG protein.</p>

Huangqi decoction inhibits cholangiocyte proliferation and transdifferentiation in cholestatic liver fibrosis induced by BDL in rats / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To elucidate the antifibrotic mechanism of Huangqi decoction in rats with BDL-induced cholestatic liver fibrosis.</p><p><b>METHODS</b>Liver fibrosis model was induced by ligating the common bile duct (BDL) in rats. Sham-operation was performed in control rats. The BDL rats were randomly divided into two groups: the BDL group and the Huangqi decoction group. Huanqi decoction was given intragastrically for 4 weeks. At the end of the fifth week after BDL, animals were sacrificed.</p><p><b>RESULTS</b>Compared with the sham control group, mortality rate in BDL group was 33.3% and incidence rate of ascites was 90%, and hepatic function was abnormal in most of the rats in BDL group. The number of Hepatocytes was decreased and the number of cholangiocytes significantly increased in BDL group. In addition, Hyp content of liver tissue and protein expression of CK 7 and a-SMA were significantly increased. Immunostaining indicated that CK 7 and a-SMA were co-localized in BDL group. These changes were markedly suppressed by the Huangqi decoction.</p><p><b>CONCLUSIONS</b>These observations suggest that Huangqi decoction can inhibit cholangiocyte proliferation and cholangiocyte transdifferentiation.</p>

Syndrome identification of CCl4 induced liver fibrosis model rats based on syndrome detecting from recipe used / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To further demonstrate the cognition upon the prominent effect of Yiguanjian and Xiayuxue Decoction on the CCI4 induced rat model of chronic liver fibrosis which have been verified in the previous studies. From the viewpoint of detecting TCM syndrome by recipe used, through this cognition the pathological features of the liver injury model manifesting a syndrome of "Gan-yin deficiency with blood stasis obstructing collaterals" were further explored.</p><p><b>METHODS</b>Wistar rats were randomly divided into the normal group, the model group, and the three medicated groups. All rats, except those in the normal group, were made into chronic liver injury model by subcutaneously injecting CCI4 for 12 weeks. Medication for the three medicated groups began from the 9th week after modeling, with oral administering of Yiguanjian (YGJ, a recipe has been verified to be effective for liver injury and fibrosis), Liuwei Dihuang Decoction (LWDH, a recipe with effects similar to YGJ) and Yinchenhao Decoction (YCHD, a recipe functioned differently) respectively for four weeks. Rats were sacrificed at the end of the experiment, changes of hepatic function, liver pathology and hydroxyproline (Hyp) content in the liver tissue were detected, and contents of Afamin and mRNA expression of alpha-smooth muscle actin (alpha-SMA) in the liver tissue were assayed as well with Real-time PCR.</p><p><b>RESULTS</b>As compared with the normal group, the pathological figures of the chronic liver injury and fibrosis and hepatic function deterioration obviously appeared in the model rats, with the liver content of Hyp and alpha-SMA mRNA expression increased, and Afamin mRNA expression decreased significantly. In the YGJ treated group, the hepatic collagen hyperplasia and deteriorated hepatic function alleviated significantly after treatment, with content of Hyp significantly lowered, and mRNA expressions of alpha-SMA and Afamin restored to some extent (P < 0.05); the same effects on mRNA expressions of alpha-SMA and Afamin were shown in the LWDH treated group, also a decreasing trend of Hyp content (0.05 < P < 0.1), and a significant decreasing of alanine transaminase (ALT) activity was found; while in the YCHD treated group these pharmacological effects mentioned above were not observed at all.</p><p><b>CONCLUSION</b>The pharmacological effects of LWDH and YGJ were similar to some degree, which gives support to the cognition that the feature of chronic liver injury model rat induced by CCI4 is attributable to yin-deficiency sydrome.</p>

Effect of xiaochaihu decoction and different herbal formulations of its components on cytokines of carrageenan induced pleuritis in rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the anti-inflammatory effect and possible mechanism of Xiaochaihu Decoction (XCHD) and the different herbal formulations of its components.</p><p><b>METHODS</b>Rat model of pleuritis was established by thoracic injecting 0.2 ml of 1% carrageenan. Rats in the treated groups were medicated with XCHD (11 g/kg) and the different herbal formulations of its components respectively as follows: thorowax root-scutellaria root (A, 3.5 g/kg), fresh ginger-pinellia tuber (B, 3 g/kg), ginseng-licorice root-jujube (C, 4.5 g/kg), A + B (6.5 g/kg), A + C (8 g/kg) and B + C (7.5 g/kg), and those in the normal group and the model group were given equal volume of instilled water, by way of gastrogavage for successive 5 days. Modeling was performed 2 hrs after the last medication. The amount of pleurorrhea, and leucocyte (WBC), marrow peroxidase (MPO), tumor necrosis factor alpha (TNF-alpha) and interleukin-1beta (IL-1beta) in pleurorrhea, and serum level of interleukin-8 (IL-8) were measured 12 hrs after modeling.</p><p><b>RESULTS</b>As compared with the model group, all the parameters measured were lower in all the treated group (P < 0.05) , and IL-1beta content in pleurorrhea in the XCHD group and Group A, B and C were significantly lower (P < 0.01).</p><p><b>CONCLUSION</b>XCHD and the different herbal formulations have obvious anti-inflammatory effect, showing certain preventive and therapeutic effect on pleuritis. Among the different formulations, the XCHD, A, B and C had better effects. The effects might be related to inhibiting the increase of cytokines as TNF-alpha, IL-1beta, and IL-8 to suppress the activation, infiltration and wandering of leucocytes.</p>