Gentianae Macrophyllae Radix is a Chinese medicinal material with unique efficacy and rich resources, which is widely distributed in northwest China. Gentianae Macrophyllae Radix contains a variety of chemical components, including iridoids, lignans, flavonoids, triterpenes, alkaloids, and other components, with anti-inflammatory and analgesic, liver protection, anti-virus, anti-tumor, immunosuppression, antihypertensive and other activities. Because of its various chemical components and wide range of pharmacological activities, it can be used as a kind of medicinal plant with great development and utilization value. With the great increase in the demand for Gentianae Macrophyllae Radix resources, the wild Gentianae Macrophyllae Radix resources are extremely shrinking. There are many medicinal sources of Gentiana Macrophylla Radix, and the medicinal parts are different, resulting in mixed sources of Gentianae Macrophyllae Radix. Not only the medicinal ingredients are unstable, the market chaos is also very serious, and the quality standard needs to be improved urgently. Based on the analysis of the present situation of Gentianae Macrophyllae Radix resources, chemical composition and pharmacological action, combined with the concept of quality markers, the quality markers of Gentianae Macrophyllae Radix were predicted and analyzed from the aspects of chemical composition and traditional medicinal properties, traditional efficacy, clinical efficacy, different compatibility and so on, in order to provide reference for the establishment of quality evaluation system of Gentianae Macrophyllae Radix.

Chlorides ; metabolism ; Diarrhea ; congenital ; Humans

Objective To study the expression of urokinase-type plasminogen activator (uPA) and its relation with expression of receptor (urokinase-type plasminogen activator receptor uPAR) in epithelial ovari- an cancer (OEC) and with the clinic prognosis.Methods Expression of uPA and uPAR protein was detected by Streptavidin-biotin-HRP in 68 cases of epithelial ovarian cancer and compared with that in 10 cases of borderline tumor,10 cases of benign tumor and 10 cases of normal tissue,and correlation between them was analyzed.The different expression groups of uPA was correlated with the prognosis of ovarian epithelial can- cer.The expression of uPA showed a correlation with short survival time (P

Abstract: Objective To establish an animal model of BALB/c mice infected with Burkholderia pseudomallei through the nose (inhalation route), provides a reliable animal model for the follow-up studies on the virulence of melioidosis and the pathogenesis of acute melioidosis. Methods　The experiment was carried out through infecting with Burkholderia pseudomallei through the nose (inhalation route). The pathophysiological response, visceral pathological damage and bacterial colonization of the mice infected with Burkholderia pseudomallei were observed by gross anatomy, histopathology and tissue homogenate count, and the biological characteristics of the mouse model of acute melioidosis were analyzed accordingly. Then we compared the physiological responses in BALB/c mice between the Burkholderia pseudomallei and non-pathogenic Burkholderia thailandensis. Results　In the model of acute nasal infection with Burkholderia thailandensis, most death happened between the 3rd to 5th day after infection, 3×105-3×106 CFU was the suitable dose for acute fatal melioidosis model of BALB/c mice, and the medium lethal dose was about 3×104-3×105 CFU. Both gross anatomy and tissue HE staining showed that abscesses or necrosis were found in the lung, spleen and liver, especially in the spleen and lung, which was positively correlated with the challenge dose. Viable bacteria was isolated from the blood, lung, spleen and liver of Burkholderia pseudomallei-infected mice, and the bacteria account colonization was related to tissue specificity. The concentration of live bacteria isolated from in the blood was the highest [Log2 value: (10.28±0.34) CFU/mL], and the organ with the maximum quantity of bacteria was the lung [Log2 value: (7.54±2.11) CFU/total organ]. It has been reported that the biological effects of Burkholderia pseudomallei and its homologous non-pathogenic Burkholderia thailandensis were similar at the cellular level, like multi-nuclear giant cell formation and active intracellular replication, while it is still unclarrified in the differences of virulence in mice. In this study, it was proved that Burkholderia thailandensis was not fatal to mice even at a high dose (8×107CFU), or detected from mice infected with it via nasal. Conclusion　We successfully established a reliable BALB/c mouse model (acute lethal model) of melioidosis via nasal infection, described its biological characteristics, and identified the different biological responses between Burkholderia pseudomallei and its homologous non-pathogenic Burkholderia thailandensis in mice.

ObjectiveTo observe the retinal function and nerve fiber layer (RNFL) thickness after laser in situ keratomileusis (LASIK). MethodsLASIK was performed in 15 cases (30 eyes) with myopia after strict preoperative examination. All examinations such as vision, correction vision, diopter, intraocular pressure, corneal thickness, ocular axis, topography scan and fundus of eye examination were performed before and 1 day, 1 week, 1 month,3 months and 6 months after operation, as well as electroretinography (ERG), visual evoked potential (VEP), optical coherece tomography at same time. ResultsThere was not significant difference in the intraocular pressure, ERG and VEP 1 day after LASIK. The thickness of RNFL decreased 1 week after LASIK (P<0.05) and recovered 1～6 months later. ConclusionLASIK does not disturb the retinal function and RNFL thickness irreversiblely.

[Objective]To investigate the molecular mechanism of abnormal platelet activation induced by platelet O2 ·- and H2O2 levels in type 2 diabetes mellitus.[Methods]The platelet parameters in patients with type 2 diabetic patients and normal controls were measured;Immunofluorescence technique was used to observe the platelet morphology changing;Flow cytometry was used to detect platelet intracellular O2 ·- and H2O2 content in two groups,then with platelets in normal controls treated with NADH/PMS system and H2O2 respectively,platelet activation positive percentage was observed. Standard Western blot analysis protocols were used to detect expression difference of Catalase and type 2 super-oxide dismutase(Mn-SOD)in platelets.[Results]The MPV in the group of type 2 diabetic patients was significantly higher than in the normal control group(P < 0.001),but there was no statisticdifference in PLT,PDW,PCT between two groups. Immunofluorescence results showed that morphology of platelets in type 2 diabetic patients changed contrast to normal group. Through flowcytometry detection,the content of mitochondrial O2·-and H2O2 of platelet in type 2 diabetic patients were obviously higher than in normal group(P<0.05),whereas no significant difference in cytoplasmic O2·-. We adopted NADH/PMS system and H2O2 to treat platelets of normal group,heightened activated positive percentage were observed which described O2 ·- and H2O2 can significantly promote platelet activation(P<0.01). Western blot results showed that expression of Catalase in platelet of type 2diabetes patients decreased,while the expression and activity of Mn-SOD had no difference.[Conclusion]It is diabetic platelet Catalase expression decreased that may lead to Diabetic platelet mitochondrial O 2 ·- and H2O2 level increased ,thus regulating aberrant activation of diabetic platelet.

AIM: To discuss the effects of phytoestrogenic-genistein on cathepsin K (CK) expression stimulated by interleukin-1α (IL-1α) in osteoclast-like cells (OCLs) . METHODS: The OCLs were isolated from tissue of human giant cell tumor of bone (GCT) . The cells treated with reagents were divided into 7 groups including control (treated with phenol red-free-DMEM), vehicle (treated with 1.2 nmol· L-1 IL- 1α), 10-10-10-6genistein, genistein+ ICI 182.780, and 17[β-estrodiol (17β-E2) group. The cells were treated with 1.2 nmol· L-1IL-1α after pre-treated with genistein or 17β-E2 for 48 h (excluded the control group) . Expression of CK wasdetermined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot in OCLs stimulated by IL-1α in the presence of genistein or 17[β-E2. RESULTS: The obvious increase of expression of CK by IL1α in vehicle group was noted in comparing with control group (P ＜ 0.01 ) . Genistein down-regulated CK gene expression stimulated by IL-1α at the transcription level in a dose-dependent manner (r = 0.68, P ＜ 0.01 ) .Genistein down-regulated CK protein expression stimulated by IL-1α also in a dose-dependent manner (r = 0.61,P ＜ 0.01 ). The effects of genistein were abrogated partly after treatment with the estrogen receptor antagonist ICI 182.780. CONCLUSION: Genistein inhibits CK expression stimulated by IL-1α partly through estrogen receptor in OCLs.

OBJECTIVE: To explore the effect of delta-opioid receptor (DOR) in electroacupuncture (EA) protecting the brain against acute ischemic injury. METHODS: Fifty-one rats were randomly divided into sham ischemia group, ischemia group, sham EA group, EA group, and EA+DOR antagonist (naltrindole) group. Transient focal cerebral ischemia (1 hour) was induced in rat brain by middle cerebral artery occlusion (MCAO) method. EA was applied on Shuigou (GV 26) and Neiguan (PC 6) for 30 min, starting immediately after the onset of reperfusion. Neurological deficit scores and volume of cerebral infarction were detected after 24-hour reperfusion. Other 12 rats were randomly divided into sham ischemia group, ischemia group, EA group and EA + naltrindole group. DOR protein expressions were assessed by Western blotting after 24-hour reperfusion. RESULTS: In comparison with the ischemia group and sham EA group, EA significantly reduced ischemic infarction and neurological deficits (P<0.05); EA significantly increased the expression of 60 kD DOR protein (P<0.05) and tended to increase that of 36 kD DOR protein (P>0.05). When naltrindole was combined with EA, the naltrindole completely abolished the EA-induced protection in ischemic infarction and neurological deficits, and also arrested the expression of DOR. CONCLUSION: EA can up-regulate DOR expression and protect the brain from ischemia-reperfusion injury.

BACKGROUND: A large amount of in vivo and in vitro experiments have confirmed that, basic fibroblast growth factor (bFGF) has been widely utilized in various tissues and cells, it can facilitate the wound healing.OBJECTIVE: To observe the efficacy and feasibility of gene gun-mediate delivery of human bFGF on the healing of deep partial thickness bum wounds.DESIGN, TIME AND SETTING: Randomized design,an observational trial was performed at the Military Central Laboratory of Changhai Hospital in the Second Military Medical University of Chinese PLA between December 2007 and October 2008.MATERIALS: SD rats of clean grade, weighing 200-250 g, irrespective of genders, ware involved in this study.METHODS: Natural human bFGF gene was recombined and optimized, then eukaryotic expression vector pCI-neo-bFGF was constructed taking pCI-neo as a vector, and transfeoted with human embryonic kidney cells 293 T cells. Dot blot and Western blot methods were utilized to determine the bFGF expression. Rat model of deep partial thickness burn wounds was processed into transgene process using gene gun technique, pCI-neo-bFGF-transfected ones served as experiment group while pCI-neo-transfected ones served as controls.MAIN OUTCOME MEASURES:Wound healing time was recorded and the efficacy was evaluated. The contents of hydroxyproline and collaganase Ⅰ in burn wound tissues were determined at 24 hours, 48 hours, 96 hours, 7 days, 10 days and 14 days following transgene process.RESULTS: the recombinant pCI-neo-bFGF was transfected with human embryonic kidney 293T cells. Dot blot and Western blot analysis have showed that, the constructed pCI-neo-bFGF expression vector could express human bFGF, and the expression of synthesized gene was remarkably higher than that of natural gene under fluorescence microscope; gene gun-mediated transgene experiment have showed that, the wound healing time was (13.00+1.31) days in the experiment group and (14.75±1.28) days in the control group, with significant differences (P＜0.05). The contents of hydroxyproline and collagenase Ⅰ reached a peak at 5 days after the injury, that is 48 hours after transfection, and then gradually decreased and maintained at a certain level. The experiment group had higher hydroxyproline levels compared with control group at different time points (P＜0.05, P＜0.01); the collagenase Ⅰ in the experiment group was notably higher than that in the control group at 48 hours and 96 hours after transfection (P＜0.01).CONCLUSION: Gene gun-mediated delivery of human bFGF can short the time of wound healing, increase the contents of hydroxyproline and collagenase Ⅰ during the healing period, accelerate the healing of deep partial thickness burn wounds.

The erythropoietin-producing hepatocellular receptor (Eph)B4 receptor is closely associa-ted with tumor growth and angiogenesis,which is over-expressed in a wide variety of tumors.Molecular probes targeted for EphB4 receptor can improve the accuracy and specificity of tumor diagnosis.A lot of molecular probes targeted for EphB4 receptor have been designed,which are expected to provide new means for the early diagnosis and therapeutics of tumors.