Objective To construct a subtractive cDNA library of genes differentially expressed in human lymphoma cell line Jurkat cells treated with phosphonoformate (PFA), and to clone genes associated with its immune regulation. Methods Using suppression subtractive hybridization (SSH) technique, the mRNA was isolated from Jurkat cells treated with phosphonoformate or 0 9 percent sodium chloride, respectively, and then cDNA was synthesized. After restriction enzyme RsaI digestion, small sized cDNAs were obtained. Then tester cDNA was subdivided into two portions and each was ligated with different cDNA adaptor. The tester cDNA, which was hybridized with driver cDNA twice and amplified by nested polymerase chain reaction (PCR) twice, was subcloned into T/A plasmid vectors to set up the subtractive cDNA library. Amplification of the library was carried out with E. coli strain JM109. The clones, which were selected randomly, were amplified by PCR, and then they were sequenced and analyzed in GenBank with Blast search. Results The subtractive cDNA library of genes differentially expressed in Jurkat cells treated with PFA was constructed successfully. The amplified library contained 46 positive clones, which contained 200- 1 000 bp of inserts. Fourteen clones were analyzed by sequencing and bioinformatics, which were identified as eleven known genes and three genes with unknown function. Conclusions The subtractive cDNA library of genes differentially expressed in Jurkat cells treated with PFA using SSH technique was constructed successfully, which brought some new clues for the study of the immune regulation mechanism of PFA

Objective To investigate the cytopathic effect of amino acid residues 86 to 175 of rotavirus nonstructural protein 4 (NSP486-175) on rat neurons and to analyze the underlying mechanism.MethodsPrimary cultured rat neurons were treated with NSP486-175 and the morphological changes induced in rat neurons were observed.Lactate dehydrogenase (LDH) activity in the culture supernatant of NSP486-175 treated-neurons was measured.Laser scanning confocal microscope was used to detect the concentration of intracellular Ca2+ labeled with Fluo-3 AM.Results Exogenous addition of NSP486-175 induced obvious cytopathic effect on rat neurons.The LDH activity in the culture supernatant of treated-neurons was stronger than that of the control group.The intensity and the distribution of fluorescence in neurons were altered after stimulation with NSP486-175.Conclusion NSP486-175 can induce the damage of rat neurons, which may be related to its role in increasing the concentration of intracellular Ca2+.This study may provide certain theoretical basis for understanding extra-intestinal spread and pathogenesis of rotavirus infection.

Objective To investigate the effect of rosuvastatin on the morphine tolerance in rats and the underlying mechanism.Methods Forty-eight male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 6 groups (n =8 each):control group (group C),morphine tolerance group (group MT),rosuvastatin control group (group RC),rosuvastatin 0.4 mg/kg group (group R1),rosuvastatin 2.0 mg/kg group (group R2)and rosuvastatin 10.0 mg/kg group (group R3).Morphine tolerance was induced by subcutaneous injection of morphine 10.0 mg/kg at 8:00 and 16:00 everyday for 5 consecutive days.The equal volume of normal saline was given in groups C and RC.Normal saline 10 ml/kg was injected through a gastric tube into stomach everyday at 30 min after subcutaneous injection of normal saline or morphine for 5 consecutive days in groups C and MT.Rosuvastatin 10,0.4,2.0 and 10.0 mg/kg were injected through a gastric tube into stomach everyday at 30 min after subcutaneous injection of normal saline or morphine for 5 consecutive days in groups RC,R1,R2 and R3,respectively.The paw withdrawal latency to nociceptive thermal stimulation was measured 1 day before (T1) and 1 day after morphine tolerance was induced (T2).The percentage of maximal possible effect (MPE) was calculated.The rats were sacrificed after the last measurement of pain threshold and the L5 segment of the spinal cord was removed for determination of the expression of extracellular signal-regulated kinase (ERK) and phosphorylated ERK (p-ERK)(by Western blot) and contents of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) (by ELISA).Results Compared with group C,MPE was significantly decreased at T2 and the expression of p-ERK and contents of IL-1β and TNF-α were increased in groups MT and R1 (P ＜ 0.05).Compared with group MT,MPE was significantly increased at T2 and the expression of p-ERK and contents of IL-1β and TNF-α were decreased in groups RC,R2 and R3 (P ＜ 0.05).There was no significant difference in the indicators mentioned above between groups R2 and R3,and in the expression of ERK between the six groups (P ＞ 0.05).Conclusion Rosuvastatin can attenuate the morphine tolerance in rats by inhibiting the phosphorylation of ERK and decreasing the level of IL-1β and TNF-α.

Objective To explore CT,MRI findings of extraskeletal mesenchymal chondrosarcoma (EMC).Methods Imaging information of all 8 cases of EMC verified by pathology were retrospectively analyzed.Results The location of lesions included lower extremity in 4 cases,forearm in 1 case,trunk in 2 cases and right lung in 1 case.The CT examination was performed in 7 cases,and 5 cases contained different patterns of ring-and-arc,granular,clump or irregular streaky mineralization.Dense calcification was detected in 3 cases,and focal in 2 cases.The nonmineralized component had slightly lower attenuation on CT scans than adjacent muscle.Four cases of peripheral located EMC demonstrated isointense on T1 WI,and mixed signal intensity on T2WI.For the cases of fine and dense calcification in 2 cases,numbers of dot-like low-intensity signals were detected resembling “pepper sign”; while for the cases of focal mineralization in 1 case,the low intensity area was located centrally in the high intensity area.Heterogeneous enhancement was found both in the calcified and uncalcified areas.One case of central located tumor exhibited low and high intensity on T1 and T2 weighted images,and nodual enhancement was observed.Conclusion EMC has several characteristic imaging features,including various mineralization pattern,enhancement of calcified area and signal intensity,which might have diagnostic value for this rare tumor.

The aim of the research is to study the prevalence and antimicrobial resistance of Campylabocter jejuni isolated from poultry in Jiangsu Province.A total of 753 samples from poultry meat and cloacal swabs were investigated,after the pure culture and the polymerase chain reaction of the mapA gene,207 isolates were examined for antimicrobial resistance by using K-B method according to World Health Organization.Results showed that all isolates performed different degree of antimicrobial resistance except meropenem,gentamycin,kanamycin,florfenicol and fosfomycin,the resistance rates of 194 strains to trimethoprim,norfloxacin,ceftriaxone and tetracycline were 100％,84.02％,80.9％ and 79.4％ respectively,1 strain isolated from Xuzhou was resistant to 92.6 ％ antibiotics.The multi-drug resistance appeared and the advantage of drug-resistant spectrum was LIN/CTX/CRO/NOR/CIP/T/TE,the resistant type focused on 40％-60％.The research provided evidence for surveillance of the antimicrobial resistance of C.jejuni and highlighted the need to employ more prudent use of critically important antimicrobial.

Objective:To prepare for mAb of progesterone receptor. It would provide support for the immunohistochemistry behind. Methods:Target gene connected together with a carrier by seamless cloning method. The target protein that expression by inducing was collected. And with cell fusion method , the monoclonal antibodies were preparation. Then the mAb were detected by IHC. Results: The mAb ( clone 7C7 ) was detected and it found positive for the breast, uterine fibroid tissue, showed negative in colorectal cancer tissue, smooth muscle tissue, the goal of the claim were achieve. Conclusion: Finally, we found the method that prepare for mAb was far beyond our imagination. The result of IHC on different samples about mAb(7C7)obtained compliance with an-ticipation. Study on the difference between the PR-A and PR-B had significance.

Objective To explore the therapeutic effects of the folic acid,vitaminB12on Helicobacter pylori(Hp)-negative patients with chronic atrophic gastritis(CAG).Methods we enrolled 67 patients who were diagnosed as CAG of Hp-negative from the First Affiliated Hospital of Xi′an Medical University.The patients were divided into control group and treatment group. Control group were given routine treatment(pepsin tablets), treatment group were given routine treatment and folic acid,vitaminB12. Then respective compared the folic acid, vitaminB12,clinical symptoms scores,gastroscopic scores and histopathological scores in the two groups before treatment and after treatment of 12 weeks and 24 weeks.Results After treatment of 12 weeks,there were signifi-cant differences in the gastroscopic scores and histopathological scores(activity),folic acid,vitaminB12status (P<0.05);no significant difference existed in clinical symptoms scores,histopathological scores(chronic inflam-mation,atrophic,intestinal metaplasia)between the two groups(P > 0.05). After treatment of 24 weeks,the differences of clinical symptoms scores,gastroscopic scores,histopathological scores(chronic inflammation, activity,atrophic,intestinal metaplasia)and folic acid,vitaminB12status were significant(P < 0.05);no significant difference existed in histopathological scores(intestinal metaplasia)between the two groups(P >0.05). Conclusion Folic acid and vitaminB12can improve the clinical symptoms and histological situation of the Hp-negative patients with CAG,worthy of further popularizing in clinic.

Objective To determine the effect of 2 transforming growth factor β1 (TGF-β1) short hairpin RNA (shRNA) expression plasmids (pcDU6-A1-A2 and pcDU6-B1-B2) on proliferation, TGF-β1, connective tissue growth factor (CTGF), and fibronectin (FN) expression induced by human serum albumin (HAS) in HK2 cells. Methods A vector plasmid containing the TGF-β1 shRNA was generated. An HK2 cell line was used in the study. The 2 TGF-β1 shRNA expression plasmids were transfected into cultured HK2 cells by lipofectamine 2000. Cellular proliferation was assessed by tetrazolium salt colorimetry. The semi-quantitative reverse transcriptive PCR was performed to detect the expression of TGF-β1,CTGF, and FN mRNA. Levels of TGF-β1 and FN protein were measured with a sandwich enzyme-linked immunosorbent assay. Results After treating with 5 g/L HAS for 24 hours in HK2 cells, cellular proliferating capacity increased significantly (P＜0.05). The expression levels of TGF-β1, CTGF, and FN mRNA were upregulated in HK2 cells stimulated by 5 g/L HAS, and levels of TGF-β1 and FN protein in the culture supernatant increased (P＜0.05). The introduction of pcDU6-A1-A2 and pcDU6-B1-B2 resulted in significant reduction of cellular proliferation activity, and the expression levels of TGF-β1, CTGF, and FN mRNA were downregulated (P＜0.05). Levels of TGF-β1 and FN protein in the culture supernatant decreased (P＜0.05) after 12 or 24 hours of TGF-β1 shRNA transfection into HK2 cells There was no significant difference in the expression levels of TGF-β1, CTGF, and FN mRNA between the 2 pcDU6 vector plasmid mediated TGF-β1 shRNA groups (P＞0.05). Conclusion pcDU6 vector plasmid mediated TGF-β1 shRNAs could obviously inhibit the expression levels of TGF-β1, CTGF, FN and cellular proliferation stimulated by HAS in HK2 cells, which may be related to the mediation of TGF-β1.

Objective To examine the sleep status and relevant psychosocial factors in patients with colorectal cancer before surgery and improve their sleep quality. Methods A cross?sectional survey method was used. Participants were 107 cases of patients with colorectal cancer from the Department of Anorectal Surgery in The First Hospital ,which is affiliated with China Medical University. The Athens insomnia scale(AIS),Ham?ilton depression scale(HAMD),Hamilton anxiety scale(HAMA),perceived social support scale(PSSS),self?esteem scale(SES),medical cop?ing questionnaire(MCMQ),memorial University of Newfoundland scale of happiness(MUNSH),Eysenck personality questionnaire(EPQ),and Wong?Baker face scale were used to assess preoperative sleep in patients with colorectal cancer and related psychosocial factors. Results Among the 107 cases,there were 24 cases of insomnia,with an insomnia incidence of 22.43%. No statistical differences were found in demographic charac?teristics and clinical characteristics(P>0.05). According to the degree of insomnia,there were significant differences between groups(P<0.05) for depression,anxiety,social support,avoidance,yield,happiness,and EPQ?N. Before the surgery,the degree of depression,anxiety,and EPQ?P had a significant positive effect on the degree of insomnia (P< 0.05). Conclusion The degree of insomnia before surgery in patients with colorectal cancer is closely associated with depression,anxiety,coping styles,social support,and personality characteristics.

Objective: To investigate occupational health status about radiation workers from township hospitals in Gansu, China, and to explore the dose-effect relationship of exposure dose of radiation workers. Methods: A total of 390 X-ray workers from township hospitals in Gansu Province and 450 X-ray workers from the county/district-level hospitals in the county/district where the township hospitals were located were divided into research group and control group. Their workload, radiation protection condition, and occupational health monitoring data were used for impact assessment and risk estimation. Results: The annual workload of radiation workers from township hospitals was 845.33±361.622 people, and that of radiation workers from county/district-level hospitals was 1967.74±762.055 people; there was a significant difference in annual workload between the two groups of subjects (P<0.05) . Indices such as detection rate of lens opacity, detection rate of abnormal platelet count, red blood cell count, and hemoglobin count, micronucleated lymphocyte rate, and chromosome aberration rate were significantly different between the two groups (P<0.05) . Conclusion Radiation workers from township hospitals suffer from certain radiation damage.