Objective To test whether sodium arsenite can induce in vitro reverse mutation of Salmonella typhimurium histamine-auxotroph mutant. Methods Ames test was carded out with Salmonella typhimurium strains TA97,TA98,TA100 and TA102 by standard method with or without the liver microsomal enzyme activation system (+S9,-S9). Results At concentrations of sodium arsenite from 500.00 to 5000.00 μg/plate, no colonies were seen on the plates of TA97,TA98,TA100 or TA102, with or without the presence of S9. At concentrations of sodium arsenite of 0.01,0.10,10.00 μg/plate and with the presence of S9, twice as many colonies grew on the plates of TA102 as the negative control(P<0.05). Without S9 activation,twice as many colonies grew on the plates of TA100 as the negative control(P<0.05)at concentrations of sodium arsenite of 1.00,10.00 μg/plate(P<0.05). The reverse mutation colonies induced by sodium arsenite in TA98 strain were twice as many as negative control group at concentrations of 0.01,0.10 μg/plate(P<0.05). There was no obvious increase of the strain clones in the other(P0.05). Conclusions With and without S9 activation, the doses of 500.00,5000.00 μg/plate sodium arsenite resulted in a toxic effect and a reduction of the revertants among the strain. At concentrations of 0.01～10.00 μg/plate, sodium arsenite exhibited mutngenesis effects.

Objective To investigate the effects of glutamine (Gln) on proliferation and survival of small cell lung cancer H446 cells, and further to explore the potential mechanism. Methods The proliferation of H446 cells was detected at different time points (0, 24, 48, 72 and 96 h) by CCK-8 assay in Gln (+) group and Gln (-) group, and an optimal time was selected. Under the optimal time, Annexin V-FITC/PI staining, CellTiter-Glo? assay kit and flow cytometer were used to detect cell survival, cellular adenosine triphosphate (ATP) and reactive oxygen species (ROS) levels. Gln (-) group was used as the control group, under the condition of Gln deficiency, cellular ATP, cell proliferation and survival were detected after adding oxaloacetic acid (OAA) or dimethyl-α-ketoglutarate (DM-αKG). Gln (-) group was used as the control group, cellular ROS, cell proliferation, colony and survival were detected after treated with ROS scavenger N- acetyl cysteine (NAC). With different concentrations (0, 2, 5, 10 μmol/L) of glutaminase inhibitor BPTES, the optimal concentration was selected through the colony assay. The cellular ATP and ROS levels and cell proliferation were detected under the optimal concentration. H446 cells were treated with bis-2-(5-phenylacetamido-1,2,4-thiadiazol-2-yl) ethyl sulfide (BPTES), ROS inducer hydrogen peroxide (H2O2) or the combination of them, and cell survival ratio was compared between two groups. Results The proliferation levels of H446 cells at 24, 48, which were decreased most significantly in 72 h in Gln (-) group. When 72 h was used as the optimal time, the cell survival ratio and ATP level were decreased, and the ROS level was increased, in Gln (-) group compared with those of Gln (+) group (P<0.05). There was a higher survival ratio in H446 cells in Gln (-)+OAA group and Gln (-)+DM-αKG group than that of Gln (-) group (P<0.05), but there were no significant differences in cell proliferation and ATP levels between Gln (-) group, Gln (-)+OAA group and Gln (-)+DM-αKG group. The ROS level was reduced, the cell proliferation, colony level and survival ratio were increased in Gln (-)+NAC group compared with those of Gln (-) group (P<0.05). Cloning assay showed that 10μmol/L was the optional concentration. Under this concentration, the proliferation and ATP level were decreased in Gln(+)+BPTES group (P<0.05), and cellular ROS level was up-regulated compared with Gln(+) group. The survival ratio was significantly lower in BPTES+H 2O2 group compared with BPTES (+) group or H2O2 (+) group. Conclusion Glutamine deficiency inhibits the proliferation and survival ratio of H446 cells through enhancing ROS level. BPTES and H2O2 show synergistically inhibitory effect on the survival of H446 cells.

Objective To investigate the effects of liver transplantation on function and morphological changes of spleen in hepatic cirrhosis rats.Methods Hepatic cirrhosis model was established in rats by subcutaneous injections of carbon tetrachloride.Liver transplantation model was established with two-cuff technique.The portal vein pressure,and morphological changes of spleen were observed before and after liver transplantation in hepatic cirrhosis rats.The serum tuftsin level before and after liver transplantation were detected.Results Before liver transplantation,PVP was increased from 6.28 mmHg to 14.03mmHg(P

Objective To study the correlation between pulse pressure(PP) and cardiovascular structure and function in aged patients with essential hypertension. Methods Forty-eight aged patients with essential hypertension were enrolled into the study and divided into the following two groups by the mean value of PP: PP

For rapid screening, we constructed two levels pools (primary and secondary pools) of the bacterial artificial chromosome (BAC) library of Chinese fine wool merino sheep. The primary pools were based on the individual 384-well microtiter plate and were prepared with a three-dimensional pooling scheme. Three dimension (plate, row and column) pools were made for each. The secondary pools were based on the entire BAC library. We developed a PCR based strategy to identify positive BACs from sheep BAC library. First, we analyzed secondary pools DNAs, according to the result, we analyzed correlative primary pools. It was one-step screening (66 PCR reactions) that we could screen a single positive clone from 74 000 BACs by our method, or three-step screening (less than 100 PCR reactions) could screen more clones. By one-step screening (66 PCR reactions), we screened successfully a positive clone 373D13 with polymorphism marker BF94-1.
Animals ; Chromosomes, Artificial, Bacterial ; genetics ; Gene Library ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; Sheep ; genetics

In order to investigate the biological effects of the VP22?-mE6?/mE7 built-in adjuvant fusion protein vaccine on the tumor associated with HPV-16 infection.HSV-1 VP22? and HPV-16 mE6?/mE7 genes were cloned,and the pET28a-VP22?-mE6?/mE7 recombinat prokaryotic expression vector was constructed.Vector was transformed into Rosetta(DE3)E.coli string and expressed under the induction of IPTG.The re-naturalized protein was then purified via Ni2+ affinity adsorbent column and identified by SDS-PAGE and Western blot.Purified protein was immunized BalB/C and C57BL/6 mice to evaluate the immunogenicity and anti-tumor activity.The expressed recombinant protein formed as inclusion body with a prediction MW about 34kDa and contained approximately 45% of total somatic protein.The VP22?-mE6?/mE7 immunization induced higher titer of specific IgG against HPV,higher level of lymphocyte proliferation and better effect on suppressing HPV16 positive TC-1 tumor growth than the mice immunized with mE6?/mE7 alone.The results showed that the recombined built-in adjuvant vaccine could induce specific cellular immune response in vitro and inhibit the TC-1 tumor proliferation in vivo,that would be a foundation for further studies and developments of inner adjuvant vaccines.

Objective To investigate the distribution characteristics of methicillin-resistant Staphylococcus aureus (MRSA)in a children's hospital,and provide basis for the prevention and control of MRSA infection in children. Methods Children who admitted to a children's hospital from 2011 to 2015 were analyzed retrospectively,clinical data of children,isolation of pathogens,types of specimens,and healthcare-associated infection(HAI)status were analyzed.Results From 2011 to 2015,a total of 911 children isolated Staphylococcus aureus (SA,1108 positive specimens),494 of whom isolated MRSA (599 positive specimens),54.23% of children isolated MRSA(isolation rate of specimens was 54.06%);there was no significant difference in the isolation rate of MRSA between children of different genders(P > 0.05);isolation rate of MRSA in different age groups was statistically significant(P <0.05).Isolation rates of MRSA from blood,puncture fluid,secretion,and pus were 68.97%,66.00%,55.81%, and 54.47% respectively.Isolation rate of SA and MRSA increased from 0.61% and 21.74% in 2011 to 1.40%and 75.59% in 2015 respectively,difference were both significant(both P <0.05).Incidence of SA and MRSA in-creased from 0.198% in 2011 to 2.697% and 2.119% in 2015 respectively,both showed an upward trend year by year(both P <0.05).Conclusion Isolation rate of MRSA and incidence of HAI in this children's hospital increased year by year,it is necessary to intensify management,use antimicrobial agents scientifically and rationally,timelyperform disinfection and isolation,so as to curb the emergence and spread of MRSA in hospital settings.

Objective To study the effects of Yiqi Huayu Jiedu Prescription on the migration ability of MHCC97-H and the expressions of CXCL12, CXCR4 and CXCR7; To discuss its relevant mechanism of action. Methods Setting Sorafenib as a positive control, CCK-8 method was used for determining the effects of Yiqi Huayu Jiedu Prescription on the cell proliferation of MHCC97-H and the optimum concentration. Scratch assay was used to observe the migration ability of MHCC97-H. The protein expressions of CXCL12, CXCR4 and CXCR7 were detected by Western blot after 24 h of medicine intervention. Results Yiqi Huayu Jiedu Prescription and Sorafenib can inhibit the cell proliferation of MHCC97-H , and the inhibitory concentration was 0.095 g/mL and 10 μmol/mL at 24 hours. Yiqi Huayu Jiedu Prescription can inhibit migration ability of MHCC97-H. The protein expressions of CXCL12, CXCR4 and CXCR7 in hepatocellular carcinoma cells decreased after the action of Yiqi Huayu Jiedu Prescription. Conclusion Yiqi Huayu Jiedu Prescription can inhibit MHCC97-H cell proliferation and migration, which may be realized by down-regulating chemokine axis of CXCL12/CXCR4/CXCR7.

Objective To establish the proposals of clinic and rehabilitation pathway for spine and spinal cord injury. Methods The experiences in clinic and rehabilitation treatment fields, the flow path of admission and the costs of hospitalization for spine and spinal cord injury patients in Chinese Rehabilitation Research Center were retrospectively summarized and analyzed. Related data in International Classification of Functioning, Disability and Health (ICF) core sets were also referred. Results The present proposals were applied to patients suffered from thoracic, lumbar or sacral spine injury with spinal cord injury. The proposals consisted of two parts: the surgical interventions in acute stage (within 2 weeks after injury) and clinic and rehabilitation treatments of spinal cord injury in sub acute and later stages (2 weeks to 6 months after injury). The criteria of surgical interventions in acute stage materialized the core therapies such as reconstruction of spine stability and neural decompression, etc. The criteria of treating spinal cord injury in subacute and later stages demonstrated the characteristic of combined therapy in which rehabilitation was the central element. Conclusion The establishment and implement of the present pathways were based on the time course of disease development and the staging feature of neural recovery and treatment after spinal cord injury. The purpose of this work is to improve the quality of medical service and reduce medical consumption.

Objective To explore the effect of gene mutations of arsenic transport proteins-muhidrug resistance-associated proteins(MRP1 and MRP2)on phenotype of endemic arsenic poisoning.Methods Two hundreds and thirty-nine rural residents in 3 villages of Shuocheng Region,Shanxi Province were interviewed and examined by simple random sampling who had been lived there for 20 yearn at least.All the objects were divided into two groups on the basis of clinical examination with"The Standard Diagnosis of Endemic Arsenic Poisoning" (WT/S 211-2001):subjectives with skin lesion as a arsenic poisoning group and without skin lesion as a control group. One hundred and ninety-three blood samples were collected from each participanL Seventy-five arsenic poisoning cases and 118 controls were detected the gene mutations in the 2,17,23 exons of M RPI and the 10,18,31 exons of MRP2 by PCR-single strand conformation polymorphism (PCR-SSCP) and compared by multivariate Logistic regression model. Results Seventy-five cases and 164 controls underwent questionnaires. Age[ (58.85±11.26) vs (45.73±11.92),OR = 3.378,P ＜ 0.05],gender[male,57.3%(43/75)vs 27.4%(45/164),OR = 3.553,P＜ 0.01 ],smoking[46.7%(35/75) vs 21.3%(35/164),OR = 3.225,P ＜ 0.01 ],drinking[ 17.3%(13/75) vs 8.5% (14/164),OR = 1.836,P ＞ 0.05],vegetable and fruit intake[5.3%(4/75) vs 9.1%(15/164),OR = 0.560,P ＞ 0.05],egg and meat intake[34.7%(26/75) vs 30.5%(50/164),OR = 1.210,P ＞ 0.05],exposure of pesticide [41.3%(31/75) vs 29.3%(48/164),OR = 1.864,P ＜ 0.05] were tested by Logistic regression model. There was no gene mutation detected in the 23 exon of MRP1 and the 18 exon of MRP2. The gene mutations frequencies of the 2 exons of MRP1 in arsenic poisoning and control groups were 8.00% (6/75) and 5.93% (7/118),respectively;they were 13.33%(10/75) and 8.47%(10/118) of the 17 exons of MRP1,respectively;they were 22.67%(17/75) and 18.64%(22/118) of the 10 exons of MRP2,respectively;they were 5.33%(4/75) and 2.54%(3/118) of the 31 exons of MRP2,respectively. There was no significant difference between two groups(x2 = 0.312,1.165,0.460, 2.794,respectively,all P ＞ 0.05). After age,gender,smoking,drinking,nutritional level and exposure of pesticide being adjusted by multivariate Logistic regression model,there was no significant difference between two groups (OR = 0.803,1.892,2.388,1.098,respectively,all P ＞ 0.05). Conclusions The gene mutations of 2,17,23 exons of MRPI and the 10,18,31 exons of MRP2 may have no effect on the phenotype of endemic arsenic poisoning.