In this study,the intestinal microbial flora diversity of adult and young African lions in the same breeding environment was detected by PCR-DGGE technique.Total bacterial DNA was extracted and 16S rDNA V3 region was amplified,then conducting PCR-DGGE.Subsequently,the specific bands of DGGE were cloned and sequenced.The bacterial species were identified by comparing the sequence through BLAST.The results indicated that the intestinal microbial flora of adult African lions includes Clostridium,Lachnospiraceae bacterium,Anaerovorax,Lactococcus,Peptostreptococcus and Blautia.While the intestinal microbial flora of young African lions is lesser,most bacteria are common to adult and young lions,such as Bacteroidetes bacterium and rumen bacterium.The UPGMA clustering analysis of the DGGE fingerprint showed the similarities of the bacteria structures between adult and young African lions were only 34％.These results revealed that the intestinal microbial flora has significant difference in different stages of African lions.This study lays a foundation for the development of microecological agents in different growth stages of wild animals.

Objective To investigate the role of GABAAα3and GABAB receptors in the ventrolateral periaqueductal gray in the development of paw acute pain in rats.Methods Twelve male SD rats, weighing 280~320 g, were randomly divided into two groups: normal saline group (group NS), formaldehyde-induced pain group (group F), 6 rats in each group.In group F, rats were subcutaneously injected with 2% formaldehyde 50 μl into the ventral surface of right hind paw to induce periphery inflammatory pain.In group NS, rats were subcutaneously injected with normal saline into the ventral surface of right hind paw.Mechanical threshold was assessed using von Frey hairs for every ten minutes.The rat pain behavior scores were recorded for every five minutes.The thickness of skin and skin temperature were recorded for every fifteen minutes.Results Mechanical hyperalgesia were induced in group F after formalin injection into right hind paw.Compared with group NS, rat pain behavior scores were increased significantly in group F at all time points after injection, mechanical threshold were decreased significantly in group F at 10-60 min after injection, the temperature of the skin and the skin thickness were increased significantly in group F at 15-60 min after injection (P<0.05), the levels of the expression of GABAAα3 and GABAB were significantly increased in group F (P<0.05).Conclusion GABAAα3and GABAB receptors mediates formalin-induced hyperalgesia at ventrolateral portion of the PAG (vlPAG) of rats.

Objective To investigate the epidemic factors of human brucellosis in Qiqihar from 2008 to 2014 so as to provide a scientific basis for prevention and control of the disease.Methods Descriptive epidemiologic method was used to analyze the monitoring results of human brucellosis in Qiqihar between 2008 and 2014.Indexes observed were:the incidence rate,the distribution of gender,age,occupation,area and time.Results From 2008 to 2014,there were a total of 15 003 cases of brucellosis covering all counties.The average incidence rate was 38.85/100 000.The average incidence rate of Meilisi Daur District was the highest,178.93/100 000.Cases occurred year around and peaked in March-July.The average incidence rate of male and female brucellosis was 57.53/100 000 and 19.35/100 000,respectively,and male patients were more than female (x2 =3 658.973,P ＜ 0.05).Most patients were in the 41-50 years old age group,and the number of patients was 4 386.The major occupation was farmer (herdsmen),which acount for 93.96％ (14 097/15 003).Conclusions Qiqihar is the epidemic area of brucellosis and has a higher incidence.Governments should pay nore attention to the epidemic of brucellosis.Health education and intervention measure for high-risk population should be strengthened and the professional team should be reinforced so as to reduce the incidence of brucellosis.

OBJECTIVE: To study signal transduction pathways in cultured rat hepatocytes in the high nitric oxide (NO) environment of hepatitis. METHODS: NO levels were assessed by measurement of its stable oxidative products nitrite (NO2(-)) and nitrate (NO3(-)) using the Griess method with or without thiols (GSH or L-Cys). Rat hepatocytes were incubated with Sodium Nitroprusside (SNP) to produce a high NO environment and the intracellular cGMP and s-nitrosoglutathione (GSNO) in the culture media were measured using radioimmunoassay or with the MTT assay absorbed at 334nm respectively. RESULTS: After incubation of 1.543 mmol/L SNP for 30 minutes 0.63+/-0.06 mmol/L and at 25 minutes 0.98+/-0.11 mmol/L of NO was released in containing 25 mmol/L GSH and L-Cys condition. The levels of both cGMP and GSNO were significantly increased (compared with control P<0.05) in a dose related manner. CONCLUSION: Signal transduction of cultured rat hepatocytes in a high NO environment could be a cGMP-dependent as well as a non-cGMP-dependent pathway.

OBJECTIVE: To establish a hepatocyte immunotoxicity model for screening of liver protective medications. METHODS: Cytotoxicity was induced by coincubating BCG-pretreated rat hepatocytes in vivo and with 10 mg/L LPS in vitro. Biphenyldimethylesterate (DDB), malotilate(MLT), silybin(SB) and glycyrrhizin (GRZ) were coincubated along with LPS to prevent the hepatocyte injury and verify the applicability and reliability of the model. AST, LDH and nitric oxide (NO) were measured in both the serum and supernatant. The liver and spleen index were calculated and the liver histopathologic changes were examined microscopically. RESULTS: Supernatant AST, LDH and NO in the BCG combined LPS group were increased in comparison with the control group (P<0.01). This increase was attenuated by the addition of DDB, MLT, SB and GRZ (P<0.05). The serum AST, NO and liver and spleen index were also increased significantly compared with the control group (P<0.01). Microscopic exam revealed serious histopathologic changes in the BCG combined LPS group. Hepatoxicity with associated liver enzyme elevation but histopathologic changes were attenuated by DDB, MLT, SB and GRZ. CONCLUSION: BCG combined LPS-induced hepatocyte immunotoxicity in an in vitro rat model may be a useful technique to assess the effectiveness of liver protective medications.

Objective To explore the effect of N-acetylcysteine nano-carbon (ACNAC) on antioxidative capacity in non-alcoholic steatohepatitis rats.Methods Rats non-alcoholic steatohepatitis model was established by means of high-fat diet in 12 weeks.Then the rats were treated with different doses of ACNAC (800 mg/kg,400 mg/kg and 200 mg/kg) and YISHANFU at the dose of 0.692 mg/kg,NAC at the dose of 400 mg/kg in astrolavaging perday.After 8 weeks,serological levels and liver tissue of SOD,MDA,GSH,GSH-PX in all groups were measured.Pathological changes of liver tissues under light microscope were observed.Results Compared with the high-fat group rats,the groups of medium dose and low dose of ACNAC in the serological levels of ALT,AST were decreased significantly,even lower than the group of YISHANFU.TCHOL,TG,LDL-C and CR were no statistical significance but decresed in date, and also lower than the groups of YISHANFU and NAC.The level of liver tissue in SOD,GSH,GSH-PX,the group of ACNAC were higher than the high-fat group's,and the level of MDA was lower than that.Conclusion Dose-related N-acetylcysteine Nano-carbon could enhance antioxidant capacity of NASH rats.

Objective To investigate the incidence rate, pathologic changes, mechanisms and diagnostic methods in renal bone disease.Methods The blood levels of carboxyterminal parpthyriod hormone (C-PTH), 1,25(OH)2D3, calcium and phosphate, aluminum in serum and bone tissue were measured. The bone biopsy and bone scan with 99m technetium methylene diphosphonate (99m TC-MDP) were performed in 51 uremic patients.Results One hundred per cent of the patients had varying degree of pathologic changes in bone, in which 50.9% of the patients presented high-turnover bone disease, 7.8% of the patients presented lowe-turnover bone disease and 41.8% of the patients had mixed-type bone disease. The levels of serum C-PTH were predominently high in high-turnover bone disease while the levels of serum 1,25(OH)2D3 were significantly decreased in low-turnover bone disease. There was a high positive rate for the diagnosis of renal bone disease by bone scan with 99m TC-MDP.Conclusions The examination of bone pathology is the most valuable method for the diagnosis of renal bone disease. Bone scan with 99m TC-MDP has reference value when clinical conditions do not allow to make bone biopsy.

OBJECTIVETo study the diagnosis of vasovagal syncope (VVS) in children with postural orthostatic tachycardia syndrome (POTS).

METHODSThe clinical data of 57 children with POTS diagnosed by the head-up tilt test between January 2007 and December 2010 were studied retrospectively. Of the 57 children, there were 29 boys and 28 girls who were aged from 5 to 16 years (12.2±1.9 years).

RESULTSTwenty-four (42%) out of 57 children were diagnosed with VVS by prolonging the duration of the head-up tilt test (for the children with orthostatic tolerance): cardioinhibitory (20 cases), mixed (3 cases) and vasoinhibitory (1 case). The POTS children with VVS were older than those without (13.0±1.4 years vs 11.5±2.1 years; P<0.05). There were no significant differences in gender and clinical symptoms between the POTS children with and without VVS.

CONCLUSIONSSome children with POTS may have coexisting VVS, suggesting that it is important to avoid misdiagnosis of VVS by prolonging the duration of head-up tilt test in POTS children with orthostatic tolerance. There are no obvious differences in gender and clinical symptoms between the POTS children with and without VVS.

Adolescent ; Child ; Child, Preschool ; Diagnostic Errors ; Female ; Humans ; Male ; Postural Orthostatic Tachycardia Syndrome ; complications ; Syncope, Vasovagal ; diagnosis ; Tilt-Table Test

OBJECTIVETo determine the role of recipient lymphopenia state in the expansion and function of leukemia specific cytotoxic T lymphocytes (CTLs).

METHODSC57BL/6 mice were induced to lymphopenia with 6 Gy total body irradiation. Spleen T cells or leukemia specific T cells from EGFP+ transgenic C57BL/6-EGFP mice were adoptively transferred by intravenous injection. The mice were challenged subcutaneously with 1 x 10(6) FBL3 leukemic cells at day 2 after irradiation. The peripheral WBC count, percentage of EGFP+ cells, subsets of T cells and tumor sizes were monitored.

RESULTBoth of the spleen T cell and leukemia specific CTL proliferated efficiently with the percentage of EGFP+ cells of 28. 81% and 42.24%, respectively, after infused into lymphopenic recipients. However, spleen T cells had no anti-leukemia effect regardless of its expansion. In contrast, leukemia specific CTLs showed a more rapid and extensive expansion under the condition of lymphopenia and a enhanced anti-leukemia immunity.

CONCLUSIONTransfusion of leukemia specific CTLs under lymphopenia state could be a feasible strategy to expand leukemia specific CTLs and generate favorable anti-leukemia effect in vivo.

Animals ; Disease Models, Animal ; Female ; Leukemia ; immunology ; Lymphopenia ; immunology ; Mice ; Mice, Inbred C57BL ; T-Lymphocytes, Cytotoxic ; immunology

Objective To ensure the medical security of the astronauts,new targeted strategies were adopted after summarizing the experience in Chinese astronauts rescue and medical aid at the main landing site,the specialty and characteristics of landing were analysied.Methods Search the publications about astronaut medical aid domestic and abroad,summarize the rescue and medical aid experiences from Shenzhou 5 to Shenzhou 10.In consideration of prolonged on-orbit time,the cold weather conditions at the landing zone of Shenzhou 11,new targeted strategies were presented.Results On the basis of the original helicopter emergency platform and first aid equipment,the emergency aid procedures were optimized,personal warm clothing,a heat preservation box,insulation blanket,self-heating pads and intraosseous rapid infusion system were used to ensure the medical security of astronauts in cold weather at the main landing site.Conclusions With the procedures optimized and the targeted strategies performed,the astronauts' s rescue and medical aid project was fully meet the cold and complex conditions at main landing site.