Objective To develop a simple,sensitive,and accurate method for the determination of shionone in rat plasma after administration of shionone.Methods The separation was developed by HPLC on a Waters shieldTM RP18 column (150mm × 3.9mm,5 μm) with a mobile phase composed of acetonitrile-0.05％ phosphoric acid water (98:2) at a flow rate of 1.0 mL/min.UV Detector was set at 200nm.Results The linear range of the standard curves was 0.043-1.720 μg/ml with the correlation coefficient of 0.995.The intra-and inter-day precisions were all below 10％.Conclusion The developed method can be successfully applied to the pharmacokinetic study.

Objective To study the regulatory effect of hydrogen sulfide (H2S) on endoplasmic reticulum stress in alveolar epithelial cells of rats with acute lung injury (ALI) induced by oleic acid (OA).Methods Seventy-two male Sprague Dawley (SD) rats were equally divided into control group (C group),oleic acid-induced ALI group (OA group),oleic acid-induced ALI with sodium hydrosulfide (NaHS) pretreatment group (OA + NaHS group) and sodium hydrosulfide treatment group (NaHS group).The model of acute lung injury was made by oleic acid intravenous injection in dose of 0.1 mL/kg.NaHS was injected intra-abdominally in dose of 1 ml/kg with concentration of 56 μmol/L 30 min before administration of oleic acid for pretreatment.In control groups,saline was used instead of oleic acid and NaHS in the equivalent volume.Six rats of each group were sacrificed at 2 h,4 h and 6 hours separately after modeling for observing the acute injury of lung tissue.Index of quantitative assessment of histological lung injury (IQA),wet/dry weight ratio (W/D) and H2S level of lung tissues were measured.The endoplasmic reticulum stress markers included glucose-regulated protein 78 (GRP78) and α-subunit of eukaryotic translation initiation factor-2 (elF2α) in lung tissues were measured by immunohistochemical staining and Western blot.Results At the three observation intervals,the IQA score and W/D ratio of lung tissues significantly increased in rats-after OA injected,but significantly decreased in other rats receiving both OA and NaHS.At the three intervals,the level of H2S in lung tissue significantly decreased in rats after OA injected,but significantly increased in other rats receiving both OA and NaHS.GRP78 and elF2α decreased in rats after OA injected,but increased in other rats receiving both OA and NaHS especially at 4 h and 6 h after modeling.Conclusions The findings suggested that H2S could promote the response to alveolar epithelial cell endoplasmic reticulum stress in rats with ALI resulting in attenuating the damage of lung tissue caused by oleic acid.

Objective To study the relationship between Treg cell numbers and level of TGF-β1,IL-10 in the immune microenvironment of rat liver cancer.Method 40 male Wistar rats were randomly divided into three groups,liver cancer model group (n =20),saline control group (n =10) and blank control group (n =10).Liver cancer model was established by intraperitoneal injection of DEN (100 mg/kg).Percentage of Treg cells in cancer tissues was detected by fiow cytometry and the expression level of TGF-β1 and IL-10 by immunohistochemical SABC.Results The ratio of Treg cells in CD4 + T cells was 14.32％ ± 4.84％ in liver cancer tissues,significantly higher than that in the blank control group 5.64％ ±6.10％ and the saline control group 7.95％ ±3.55％.The difference was statistically significant (F =211.279,P ＜ 0.05).The expression level of TGF-β1 and IL-10 in cancer tissues was significantly higher than the blank control group and the saline control group (FTGF-β1 =250.740,FIL-10 =152.744,all P ＜0.05).The number of Treg cells was positively correlated with TGF-β1,IL-10 level (P ＜ 0.05).Conclusions TGF-β1,IL-10 and Treg cells significantly increased in rat experimental liver cancer tissues,and there was positive correlation between Treg cells,and TGF-β1 and IL-10 in liver cancer tissues.

Objective To investigate the feature of fracture in elderly patients in Beijing Jishuitan Hospital from 2009 to 2016.Methods The data of elderly patients with fracture visiting the department of trauma emergency in Beijing Jishuitan Hospital from 2009 to 2016 were retrospectively reviewed.The data of patient number,age,gender,seasonal and circadian variation of visit and fracture site were collected and analyzed.Results Among all the 42 988 elderly patients with fractures visiting department of trauma emergency from 2009 to 2016,there were more female patients than male patients (P ＜ 0.01).In both male and female patient groups,there was a trend of increase in number of patients year by year (P ＜ 0.01).Elderly patients in the groups of 60-69 years,70-79 years and 80-89 years demonstrated a trend of increasing number from year to year notably in the 60-69 years group (P ＜ 0.01) but the total number of patients decreased as age increased.As for seasonal variation,autumn had the highest number of cases and spring the lowest (P ＜ 0.01).Most elderly patients visited emergency during 8:00-16:00 period (P ＜ 0.01).The commonest sites of fractures were at radius and femur.Conclusions Fracture of elderly patients has its unique feature in gender,age,seasonal and circadian variation of visit and fracture sites.The community should do a better planning for prevention and management of elderly patients with fractures according to their specially clinical features.

Site-directed mutagenesis method was used to introduce two desired mutations, which were confirmed by DNA sequencing, into mouse complement receptor Type II gene(MCR2). Then the constructed eukaryotic expression vectors containing wild type mouse CR2/1(wtMCR2/1), mutant type mouse CR2/1 (mtMCR2/1) and human CR2 (hCR2) cDNA were transferred into mouse SP2/0 cells by electroporation. After two-week screening by G418, the stably transfected clones were obtained. Several ways including PCR, RT-PCR, and immunohistochemistry were utilized to screen those clones with interesting genes integrated and expressed. Then Epstein-Barr virus(EBV) was used to infect these transfected cells and EBER-1 (EBV encoded RNAs) hybridization results showed that only hCR2 and mtMCR2 transfected SP2/0 cells could be infected by EBV, but positive rate of the former was much higher than the latter. This study sets groundwork for elucidating the mechanism by which EBV enters the cells and for establishing the animal model of EBV-related nasopharyngeal carcinoma (NPC).

Purpose To investigate the expression of C-C chemokine receptor 9 (CCR9) in non-small cell lung cancer (NSCLC) and to explore its prognostic value. Methods The expression of CCR9 was detected by immunohistochemistry in tumor tissues and corre-sponding adjacent normal tissues from 119 NSCLC patients. Additionally, the correlation between CCR9 expression and the clinicopath-ologic features of NSCLC and the relationship between prognostic factors and overall survival rate were analyzed by statistical methods. Results The positive expression rate of CCR9 was significantly higher in NSCLC tissues (54. 6%) than that in adjacent normal lung tissues (10. 1%) (P<0. 05). The expression of CCR9 in NSCLC was correlated with histopathologic type, lymph node status and p-TNM stage (P<0. 05). Kaplan-Meier survival analysis suggested that the positive expression of CCR9 was negatively correlated with the overall survival rate (Log-rank=9. 917, P=0. 002). Univariate analysis showed that the lymph node status, p-TNM stage and the positive expression of CCR9 made great difference to postoperative overall survival (P<0. 05). Multivariate analysis showed that CCR9 expression was an independent prognostic factor for overall survival of NSCLC patients ( RR=0. 447, 95%CI:0. 201 ~0. 993, P<0. 05). Conclusion The expression of CCR9 may predict a poor prognosis in the patients with NSCLC, so it can be used as a novel NSCLC biomarker.

Aim To study the anti-stress depression effect of Jiaweisinisan(JWSNS)and it's mechanism of N-methyl-D-aspartate(NMDA)receptor channel in hippocampus.Methods Chronic mild unpredictable stress(CMUS)was adopted to establish the rat depres-sion model.Before and after model establishing,the sucrose consumptions and the weight of rats were detected.TTC dyeing of hippocampus brain slices ex vivo were used to detect the situation of hippocampus neuron damage and the protective effect of serum containing components of JWSNS.The opening probability and the average opening time of NMDA receptor channels were detected by attach on cell patch clamp,the Ca2+ concentrations in single cell of hippocampus neurons were detected by Tillvis ION software.Results CMUS could decrease the sucrose preference and the weights of rats(P

AIM: To establish nasopharyngeal carcinoma(NPC) cell lines stable expressing NPC-derived latent membrane protein 1(LMP1) gene. METHODS: General expression vector and epithelium-specific expression vector of NPC-derived LMP1 gene were constructed by using recombinant techniques, then transfected these vectors into a poor differentiated NPC cell line named CNE-2 ,integration and expression of N-LMP1 in CNE-2 cells were detected by PCR,RT-PCR and Western blot. RESULTS: (1) General expression vector and epithelium-specific expression vector of NPC-derived LMP1 gene were constructed successfully.(2) It showed that N-LMP1 gene expressed in CNE-2 cells correctly. CONCLUSION: The first NPC cell lines which stable express NPC-LMP1 were established. The cell lines obtained will provide important basis for exploring the role of NPC-LMP1 in nasopharynx carcinogenesis.

OBJECTIVETo explore the protective effects of extract of Ginkgo biloba (EGB) in adriamycin (ADR)-induced heart failure (HF) and the mechanism of ghrelin peptide.

METHODWistar rats were randomly divided into three groups: control group, HF group and EGB group. ADR was injected in the rats of HF group and EGB group by caudal vein. After the last injection, the rats in EGB group were given intra-gastric administration of EGB solution (100 mg x kg(-1) x d(-1)). Three weeks later, cardiac function was detected; Ghrelin levels in plasma and myocardium were measured by radio-immunology assay (RIA); High energy phosphates (HEP) contents in myocardium were measured by HPLC; Myocardial gene expression of ghrelin was measured by RT-PCR.

RESULTCompared with control group, HF group had obviously decreased index of cardiac function, and these indexes such as +/- dp/dt max in EGB group were higher than those in ADR group. Plasma ghrelin level in HF group was higher than that in control group while myocardial ghrelin level was significantly lower than that in control group. Myocardial ATP content and gene expression of ghrelin mRNA in HF group were significantly lower than those in control group; Plasma ghrelin level in EGB group was significantly increased. Myocardial ATP content and gene expression of ghrelin mRNA in EGB group were significantly higher than those in HF group, and were closed to those of control group.

CONCLUSIONMyocardial energy dysfunction is an important reason of ADR-induced HF. EGB therapy can improve cardiac function and energy metabolism in HF rats, partly because it might increase the expression and production of ghrelin, which can promote positive energy metabolism.

Animals ; Disease Models, Animal ; Doxorubicin ; adverse effects ; Gene Expression ; drug effects ; Ghrelin ; metabolism ; Ginkgo biloba ; chemistry ; Heart Failure ; drug therapy ; genetics ; metabolism ; Humans ; Male ; Myocardium ; metabolism ; Plant Extracts ; administration & dosage ; Protective Agents ; administration & dosage ; Random Allocation ; Rats ; Rats, Wistar

Objective To investigate the relationship and clinical significance between the concentration of 25-hydroxy vitamin D3 [25-(OH)D3 ]in the serum and community-acquired pneumonia(CAP)in infants.Methods The study recruited 98 infants.There were 68 cases of mild pneumonia and 30 cases of severe pneumonia.The con-centration of 25-(OH)D3 in serum,IgA and alkaline phosphatase(AKP)were measured.33 infants who accepted physical examination in the outpatient department were selected as control group at the same time.Results The serum 25-(OH)D3 concentration of severe pneumonia was (21.09 ±7.32)ng/mL,mild pneumonia was (25.77 ± 8.06)ng/mL and the control group was (37.13 ±6.94)ng/mL.The differences among the three groups had statisti-cally significant differences(t =9.18,3.72,5.34,all P <0.05).The differences of serum IgA concentration among the three groups had statistically significant differences (t =5.42,3.96,8.92,all P <0.05).But serum AKP concen-tration among the three groups had no statistically significant differences(t =0.86,0.58,0.47,all P >0.05).The prevalence of Vitamin D deficiency was 40.0%,which was higher than mild pneumonia patients(20.6%)and the healthy children(9.1%)(χ2 =8.43,17.55,all P <0.05).Conclusion The serum 25-(OH)D3 concentration and IgA of CAP patients were lower than healthy children,especially in severe pneumonia cases.The prevalence of Vita-min D deficiency was significantly higher than healthy children.There were no relevance between the serum concentra-tion of 25-(OH)D3 ,IgA and AKP.25-(OH)D3 plays an important role in the development of CAP.The low level of serum 25-(OH)D3 concentration may be one of the risk factors and has correlation to the severity of pneumonia.