Objective To observe the effect of Jiakang formula on thyroid hormone and thyroid pathological changes of experimental hyperthyroid rats. Methods Forty rats were given Euthyrox daily at 75 ?g/100 g ig and randomly divided into hyperthyroidism model group, control group, middle dose Jiakang formula group, high dose Jiakang formula group, each with 10 rats, as well as 10 rats as normal group. Three weeks later, the rats were killed for observing the effect of Jiakang formula on thyroid hormone and thyroid pathological changes of experimental hyperthyroid rats. Result Jiakang formula could lower the level of FT3, FT4, T3, T4 and promote the level of TSH, also improve thyroid pathological changes, especially in high dose Jiakang formula group. Conclusions Jiakang formula had the effect of improving the function and pathology of thyroid on experimental hyperthyroid rats.

Objective To evaluate the efficacy of a two-step infusion therapy with meropenem against noso-comial multi-drug resistant Pseudomonas aeruginosa infections in elderly patients with chronic obstructive pulmonary disease.Methods 112 elderly patients with nosocomial multi-drug resistant Pseudomonas aeruginosa infections and chronic obstructive pulmonary disease were randomized into two groups:two-step infusion therapy group ( rapid first-step infusion in 30 minutes and slow second-step infusion in 2.5 hours) and traditional 30 minutes infusion therapy group.The antibiotic clinical efficacy and bacteriological efficacy were observed between two groups .Results The clinical efficacy of observation group and control group were 82.2%(46/56) vs 57.1%(32/56).The difference was statistically significant(χ2 =10.185,P<0.05);The bacteriological efficacy of observation group and control group were 75.0%(42/56) vs 46.4%(26/56),The difference was statistically significant (χ2 =10.265,P<0.05). Conclusion The clinical efficacy and bacteriological efficacy of two-step infusion therapy with meropenem against nosocomial multi-drug resistant Pseudomonas aeruginosa infections in elderly patients with chronic obstructive pulmo -nary disease were superior to the traditional therapy .

Objective To establish an indirect immunofluorescence assay for detection of murine norovirus ( MNV) .Methods Mouse leukaemic monocyte macrophage cell line RAW 264.7 cells were infected with MNV-1 and cultured for 36 hours to collect the virus and uninfected cells , and to make antigen glass slides .BALB/c mice were gavaged with MNV-1 (107 TCID50) and infected sera were collected as positive control .The serum was 1:10 diluted and used for measuring MNV antibody by immunofluorescence assay ( IFA ) .80 serum samples were tested using the two methods , IFA and ELISA, and the discrepant samples were validated by Western blotting .Results RAW264.7 cells were infected with MNV-1 for 36-48 h, showing an infection rate of 60% of the cells, and the cells infected for 36 h were preferred.IFA method was used to detect the serum with MNV-1 infection and showed that the antibody content was gradually increased at one week after infection , reaching a maximum antibody concentration at 4 weeks after infection , and maintained a stable level later .The mouse serum at four weeks after MNV-1infection was used as positive quality control . Among the 80 serum samples , 27 positive and 53 negative cases were detected by IFA method , and 32 positive and 48 negative cases were detected by ELISA .The five discrepant samples were verified by Western blotting , resulted in 3 positive and 2 negative cases . The coincidence rate of IFA was 96.0% and that of ELISA methods was 97.5%. Conclusions Basically, immunofluorescence assay can be used to detect the MNV-1 infection in mice, although false negative result may occur occasionally .IFA and ELISA detection can be selected as initial screening measures , and use Western blot assay to verify the discrepant samples .

Objective In order to establish a rhesus monkey model of p53 gene silencing, firstly we screened and determined the effective silencing targets of p53 gene at the cellular level in rhesus monkey.Methods The expression of p53 gene was detected in COS-7 cells ( derived from the kidney of the African Green Monkey, Cercopithecus aethiops).Three small hairpin RNA ( shRNA) sequences targeting rhesus monkey p53 gene were designed, analysed by bioinformatics, and inserted into lentivirus-based gene silencing constructs FUGW-TDT.The plasmids of p53-RNAi and control vector were transfected into the COS-7 cells, respectively.The suppression of p53 mRNA was detected by real-time PCR, and the changes of p53 protein expression were detected by Western blot assay.Results p53 gene expression was detected in COS-7 cells.Bioinformatics analysis showed that three gene-silencing sequences were screened which lied in the open reading frame ( ORF) region and targeted 238 -258bp, 681 -701bp, 169 -189bp of the rhesus monkey p53 mRNA.At 48 hrs after transfection of the three silencing constructs, p53 mRNA was suppressed by(87.17 ±4.03)%, ( 72.62 ±4.11)% and(76.22 ±0.98 )%, and p53 protein was suppressed by ( 84.44 ±2.18 )%, ( 71.04 ±1.18)% and ( 74.17 ±0.95 )%, respectively. Conclusions We obtained three effective target sequences showing high efficiency in p53silencing, which can be used in further studies on gene silencing in rhesus monkey.

Objective To explore the feasibility of detecting the cytokeratin 20 (CK-20) mRNA in exfoliated urothelial cells for the diagnosis of bladder carcinoma. Methods Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of CK-20 mRNA in cells collected from the urine of 45 cases of bladder cancer, 15 cases of cystitis accompanied by hematuria, 10 healthy volunteers, and 7 different cell lines, including bladder cancer cell line T24, kidney cancer 786-0 and GRC-1, breast cancer MCF-7 and MDA-MB-435, and ovary cancer SKOV 3 and 3AO. Results CK-20 mRNA expression was detected in 36 of 41 cases of bladder transitional cell carcinoma (87.80%), in 18 of the 21 GⅠ patients (85.71%), in 11 of the 13 GⅡ patients (84.62%), in 7 of the 7 GⅢ patients (100%), in 20 of the 22 T a-1 patients (90.91%), and in 16 of the 19T 2-4 patients (84.21%). Sensitivity of the method was found to be 87.80%, whereas specificity was 73.33%. In 15 patients with hematuria, there were 4 cases of false positive: 1 case of BPH, 1 case of atypical hyperplasia, 1 case of chronic inflammation, and 1 case undergoing TURP previously. CK-20 amplification band was also obtained in all of 19 cases of bladder transitional cell tumor tissues and bladder cancer cell line T24, but not in 4 patients with non-transitional cell carcinoma and 6 other tumor cell lines. No false positive cases were found in the healthy control group. Conclusion These results suggest that CK-20 might be a useful tumor marker for early noninvasive diagnosis and follow-up of bladder cancer by detecting CK-20 mRNA expression of uroepithelial cells from the voided urine specimen by RT-PCR.

Objective To research the efficiency of short time intermittent veno-venous hemofiltration (SIVVH)on acute respiratory distress syndrome(ARDS). Methods Totally 95 patients suffered ARDS between January 2014 and October 2016 in the emergency intensive care unit(EICU)were divided to SIVVH group(n =37)and control group(n = 58). The SIVVH group received traditional therapy and SIVVH therapy,and control group traditional therapy. Vital signs,oxygenation index,APACHEⅡ scores between two groups before the treat-ment were compared. Vital signs,oxygenation index,APACHEⅡ scores,time of mechanical ventilation,length of stay of EICU and 28 day mortality between two groups after the treatment were compared. Results In two groups,vital signs,oxygenation index and APACHEⅡscores before the treatment showed no significant difference (P > 0.05). In SIVVH group ,heart rate and respiratory frequency on the 24th ,48th ,72nd and 120th hour were descended significantly when compared with those in control group(P < 0.05);body temperature on the 24th and 48th hour were descended obviously when compared with those in control group(P < 0.05);the oxygenation index on the 24th,48th,72nd and 120th hour were raised(P < 0.05);APACHEⅡ scores on the 72nd and 120th hour were lower than those in control group(P<0.05). There were shorter time of mechanical ventilation and length of stay of ICU in SIVVH group when compared with control group(P<0.05),but the 28 day mortality in SIVVH group was not lower when compared with that in control group(P>0.05). Conclusions SIVVH can improve the function of lung in ARDS,decrease the time of mechanical ventilation and the length of stay of ICU,and possible becomes an important adjunctive therapy.

Objective To screen and optimize the microsatellite DNA primers of the laboratory common marmoset, analyze and evaluate the population genetic quality for the marmosets (Callithrix jacchus) introduced into the Institute of Medical Laboratory Animal science, Chinese Academy of Medical Sciences. Methods A total of 30 marmosets were randomly chosen, and their genome DNA from blood was extracted using phenol/chloroform method. The microsatellite DNA was amplified using standard polymerase chain reaction (PCR). The amplification products were tested by STR scanning after 2% agrose gel and 8% PAGE electrophoresis. The data processing and genetic analysis were completed using the Popgene1. 32 software. Results A total of 20 pairs of microsatellite loci showed genetic polymorphism, and 147 alleles were detected. The number of allele was 5 to 10, average 7. 35. The effective allele was 2. 2500 to 6. 3830, average 4. 0402. The observed heterozygosity was 0. 000 to 0. 4667, average 0. 1533. The expected heterozygosity was 0. 1424 to 0. 4350, average 0. 2506. The Shannon diversity index was 1. 2242 to 2. 0324, average 1. 5949. The polymorphic information content was 0. 5366 to 0. 8254, average 0. 7053. Conclusions The 20 pairs of marmoset microsatellite primers are genetically highly diverse and are in a Hardy-Weinberg equilibrium.

Objective To purify marmoset serum IgG, prepare and identify the antiserum and the rabbit anti-marmoset antibody IgG-HRP (horseradish peroxidase). Methods Using SDS-PAGE analysis to identify the serum IgG from HiTrapTM Protein G. The antiserum titer was determined by double immunodiffusion assay. The rabbit anti-marmoset IgG was labeled with HRP by improved sodium periodate method. ELISA and western blotting were used to evaluate the concentration and specificity of rabbit anti-marmoset IgG-HRP. Results The purity of purified marmoset serum IgG determined by SDS-PAGE was higher than 95% , and the anti-serum titer of the anti-marmoset IgG polyclonal antibody was 1∶64. The concentration of rabbit anti-marmoset IgG-HRP identified by direct ELISA was 1∶256 000, and that by western-blotting was 1∶15 000, with a strong specificity. Conclusions The IgG-HRP marker antibody is prepared and the specificity and concentration are identified by ELISA and western blotting. It reserves the resources for the detection system of marmoset pathogens and the molecular immunological testing system.

Objective To investigate the effect of the location of middle cerebral artery (MCA) occlusion on outcomes after intravenous thrombolysis with recombinant tissue plasminogen activator (rtPA) in patients with acute ischemic stroke.Methods The consecutive patients with stroke of acute MCA occlusion treated with rtPA intravenous thrombolysis within 4.5 h after onset were included.The locations of MCA occlusion were divided into either a proximal MCA segment (proximal M1 segment) or a distal MCA segment (distal M1 segment,M2 segment and more distally).Early neurological improvement was defined as National Institutes of Health Stroke Scale (NIHSS) score improvement ≥4 points from baseline or NIHSS 0 point at 24 h after thrombolysis.They were divided into a good outcome group (0-2) and a poor outcome group (3-6) according to the modified Rankin Scale (mRS) scores.Results A total of 70 patients with MCA occlusion were enrolled in the study,including 22 (31.4％) with proximal MCA occlusion and 48 (68.6％) with distal MCA occlusion;52 (74.3％) with good outcome and 18 (25.7％) with poor outcome.The proportion of atrial fibrilhtion (x2 =4.541,P =0.033),the NIHSS scores on admission (t =5.192,P =0.026) and 24 h after thrombolysis (t =5.365,P =0.024) in the proximal MCA occlusion group were higher than those in the distal MCA occlusion group.The proportion of early neurological improvement in the proximal MCA occlusion group was significantly lower than that in the distal MCA occlusion group (x2 =9.434,P =0.002),and the incidence of symptomatic intracranial hemorrhage (x2 =9.563,P =0.002)and the mortality rate within 7 d (x2 =14.491,P ＜0.001) were significantly higher than those in the distal MCA occlusion group.The time from onset to thrombolysis (t =6.346,P =0.014),NIHSS scores on admission (t =4.498,P =0.038) and at 24 h after thrombolysis (t =4.866,P =0.028),and the proportion of proximal MCA occlusion (x2 =18.710,P ＜0.001) in the poor outcome group were significantly longer or higher than those in the good outcome group.Multivariatelogistic regression analysis showed that the proximal MCA occlusion (odds ratio [OR] 14.385,95％ confidence interval [CI] 2.525-81.925;P =0.003),longer time from onset to thrombolysis (OR 12.927,95％ CI 2.624-61.748;P =0.002),higher NIHSS score at 24 h after thrombolysis (OR 3.492,95％ CI 1.027-11.880;P=0.045) were the independent predictors for poor outcome at 90 d.Conclusions There are differences in the outcomes after intravenous thrombolysis in patients with MCA occlusion at different locations.The locations of MCA occlusion,time from onset to thrombolysis,and NIHSS score at 24 h after thrombolysis,and age are the independent predictors for the outcomes after intravenous thrombolysis in patients with acute MCA ischemic stroke.

Objective To investigate the influence of moderate to severe leukoaraiosis (LA) on hemorrhagic transformation and prognosis of patients after intravenous recombinant tissue plasminogen activator thrombolysis for acute ischemic stroke and analyze influencing factors of the clinical prognosis.Methods We consecutively collected patients with acute infarct on anterior circulation (n =78) in Department of Neurology or Emergency of our hospital between January 2014 and March 2017,and all patients received intravenous thrombolysis therapy within the 4.5-hour time window.All patients processed brain MRI after intravenous thrombolysis therapy.According to the degree of LA,all subjects were classified into two groups;LA group (moderate to severe) vs no LA group (absent to mild).Clinical data were obtained and compared among patients with different grades of LA.Logistic regression analysis was used to confirm the relevant factors of prognosis 90 days after stroke.Results Among 78 enrolled patients,24 (30.8％) were classified as LA and 54 (69.2％) as no LA.In the group of LA,33.3％ (8/24) patients conducted hemorrhagic transformation,whereas 11.1％ (6/54) patients also underwent hemorrhagic transformation in the group of no LA.There was a significant difference between the two groups (x2 =5.571,P =0.018).But symptomatic intracranial hemorrhage accounted for 16.7％ (4/24) and 5.6％ (3/54) respectively in the two groups without significant difference (x2 =2.304,P =0.129).Three-month recurrence of stroke in the groups of LA and no LA was 20.8％ (5/24) and 5.6％ (3/54) respectively,also without significant difference between the two groups (x2 =3.850,P =0.050).Age ((73.7 ± 6.7)years vs (61.3 ± 10.6) years,t =6.567,P =0.012),90 d Fugl-Meyer Scale (FMS) score (92.3 ± 3.4 vs 72.9 ± 7.8,t =22.345,P ＜ 0.01) and proportion of 90 d modified Rankin Scale score 0-2 (83.3％(45/54) vs 50.0％ (12/24),x2 =9.383,P =0.002) were significantly different between the two groups.Follow-up 90-day after onset showed that the good outcome was found in 57 cases (73.1％),poor outcome in 21 patients (26.9％) and death in six cases (7.7％).The grade of LA (57.1％ (12/21) vs 21.1％ (12/57),x2 =9.383,P =0.002),silent lacunar infarction (66.7％ (14/21) vs 35.1％ (20/57),x2 =6.224,P =0.013),age ((72.8 ± 7.9) vs (61.5 ± 11.7) years,t =4.423,P =0.039),proportion of thrombolysis within 3.0-4.5 hours (71.4％ (15/21) vs 38.6％ (22/57),x2 =6.634,P =0.010),vascular occlusion size (66.7％ (14/21) vs 38.6％ (22/57),x2 =4.865,P =0.027),infarction size (52.4％ (11/21) vs 12.3％ (7/57),x2 =14.053,P =0.001) and baseline NIHSS score (16.9 ±6.7 vs 9.5 ± 4.5,t =5.426,P =0.022) were significantly different between the two groups.After adjusting for age,thrombolysis time,baseline NIHSS score,infarction size,vascular occlusion size and silent lacunar infarction,multivariate analysis revealed that moderate to severe LA (OR =4.564,95％ CI 1.199-67.724,P =0.033) was risk factor for worse outcome of patients after intravenous thrombolysis.Conclusions Acute ischemic stroke patients with moderate to severe LA have high hemorrhagic transformation after intravenous thrombolysis and may have poor 90-day FMS score.And moderate to severe LA was found to be an independent risk factor for prognosis in cerebral infarction patients with intravenous thrombolysis.