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

A novel method was proposed for the quantification of nitrite ion in aqueous samples by the combination of an advanced quantitative fluorescence model with ratiometric fluorescence indicator 2 , 3-diaminonaphthalene . The proposed method was tentatively applied to direct quantitative determination of nitrite ion in turbid environmental water samples containing both scatterers and absorbers; and its performance was thoroughly investigated and evaluated. Experimental results showed that the proposed method could realize accurate quantitative determination of nitrite ion in environmental water samples with recovery rates in the range of 90. 8%-103. 0%, which were comparable to the corresponding values of HPLC-DAD experiments. The limit of detection and limit of quantification of the proposed method were estimated to be 1 . 9 and 5. 8 nmo/L, respectively.

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 analyze the clinical characteristics and laboratory indexes of pneumonia complicated with pertussis syndrome in Suzhou area in order to provide the basis for rational treatment.Methods Children who had been hospitalized at Department of Respiratory,Children's Hospital of Soochow University during January 2012 to October 2016 were enrotled and diagnosed as pneumonia complicated with pertussis syndrome.A total of 236 cases were enrolled.Multiple pathogen detection and clinical information were collected in all patients.The subjects were divided into 28 days-＜ 3 months group,3-＜ 6 months old group,6-＜ 12 months old group and ≥ 12 months old group.The clinical data of children in each age group were retrospectively analyzed.Results Pneumonia complicated with pertussis syndrome was easy to occur within 6 months of infants,with the highest incidence in the 3-＜ 6 months group,there was a significant difference among the different age groups (x2 =231.870,P ＜ 0.05) [28 d-＜ 3 months group:5.3％ (102/1 910 cases),3-＜6 months group:5.7％ (76/1 341 cases),6-＜ 12 months group:2.0％ (36/1 762 cases),≥12 months group:0.4％ (22/5 304 cases)].The patients could become sick all the year round,with the highest incidence in summer[2.6％ (72/2 740 cases)in spring,3.7％ (96/2 611 cases)in summer,2.1％ (56/2 749 cases) in autumn,and 0.48％ (12/2 487 cases)in winter],and there were significant differences among the different seasons (x2 =62.380,P ＜0.001).Clinical symptoms were paroxysmal spasmodic cough,33.0％ (78/236 cases) of the performance with wheezing,mainly in the older than 3 months group.The incidence of fever was positively correlated with age(x2 =12.938,P ＜ 0.05).Peripheral white blood cell count and lymphocyte percentage increased,and the 3-＜ 6 months old group increased markedly.White blood cell count as high as (19.01 ± 11.99) × 109/L,the highest percentage of lymphocytes was up to 0.80,platelets were significantly increased in more than 80％ of children.The incidence of C-reactive protein (CRP) was negatively correlated with age,28 days-＜ 3 months group more prone to have high CRP.Pulmonary inflammatory pathology showed multiple pulmonary involvement by chest radiography,and it was more likely to occur in 28 d-＜ 3 months group.Viruses,bacteria and mycoplasma pneumoniae (MP) could cause pneumonia associated with pertussis syndrome,and mixed infection occurred in 56 cases.The top three pathogens were rhinovirus,MP and Streptococcus pneumoniae,and mixed infection was more prone to occur in 28 d-＜ 3 months group.Conclusion Pneumonia complicated with pertussis syndrome is easy to occur within 6 months of infants,with the highest incidence in summer.Viruses,bacteria and MP all could cause pneumonia associated with pertussis syndrome,and mixed infection could occur in some cases.The top three pathogens are rhinovirus,MP and Streptococcus pneumoniae.

Objective: To analyze the infection composition and drug resistance to Gram-negative (G^-) bacilli in children′s respiratory tract in Suzhou, in order to provide evidence for rational use of antibiotics clinically. Methods: G^- bacilli culture samples were collected from 21 561 cases of nasopharyngeal secretions from patients with respiratory tract infection admitted at the Department of Respiratory, Children′s Hospital of Soochow University from January 2007 to December 2016, including 21 246 cases in general wards, and 315 patients who were transferred to the respiratory department after treatment in the Intensive Care Unit(ICU), and the children were divided into the general ward group and the ICU group, and the pathogens were compared and the changes in bacterial susceptibility were dynamically observed between the 2 groups. Results: The primary G^-bacteria for respiratory infection was Haemophilus influenzae, followed by Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter baumannii.The detection rates of Klebsiella pneumoniae and Pseudomonas aeruginosa in the ICU group were 16.8% (21/125 strains)and 14.4%(18/125 strains), respectively, which were significantly higher than those in the general ward group [10.0%(208/2 071 strains), 9.2%(190/2 071 strains)]. The detection rates of G^-bacteria in the ICU group were 33.7%(106/315 cases), which were significantly higher than those in the general ward group [9.4%(1 997/21 246 cases)], and the difference was statistically significant(χ²=210.325, P<0.001). The rare G^-bacillus such as Stenotrophomonas maltophilia, Acinetobacter junii and Burkholderia onion were higher in the ICU group [17.6%(22/125 strains)] than that in the general ward group [6.4% (132/2 071 strains)]. The rate that of G^-bacteria with two or more mixed infection in ICU group [17.0% (18/106 cases)] was significantly higher than in the general ward group [3.4%(68/1 997 cases)], and the difference was statistically significant(χ²=47.3, P<0.05). For the mixed infection, the ICU group was mainly composed of Klebsiella pneumoniae mixed with Pseudomonas aeruginosa or Escherichia coli, while the general ward group was composed of Haemophilus influenzae mixed with Pseudomonas aeruginosa or Escherichia coli.The sensitivity of Haemophilus influenzae to Ampicillin, Sultamicillin, Cefuroxime, Cefaclor and Azithromycin decreased, and the sensitivity to Chloramphenicol, Tetracycline and Trimethoprim+ Sulfamethoxazole increased year by year, and there were statistically significant differences in different years (all P<0.05). The sensiti-vity to Escherichia coli to Ceftazidime decreased year by year, and the sensitivity to Ampicillin and Levofloxacin increased year by year, and there were statistically significant differences in different years (all P<0.05). The sensitivity to Klebsiella pneumoniae to Cefoperazone/Sulbactam and imipenem decreased, and the sensitivity to Ciprofloxacin and Levofloxacin increased, and there were statistically significant differences in different years (all P<0.05). The sensitivity to Pseudomonas aeruginosa, Cefoperazone/Sulbactam and Ceftriaxone decreased year by year, and the sensitivity to Levofloxacin increased, and there were statistically significant differences in different years (all P<0.05). The detection rate of carbapenem-resistant strains of Klebsiella pneumoniae and Pseudomonas aeruginosa showed an increasing trend, and there were statistically significant differences in different years (all P<0.05). Conclusions The primary G^-bacteria for respiratory infections is Haemophilus influenzae, G^-bacilli especially, the mixed infection of G^-bacilli, is more likely to cause severe and critical respiratory infections.The resistance rate of G^-bacteria infection in children′s respiratory tract to commonly used antibiotics is generally on the rise.

Objective To observe the therapeutic effect of endovascular embolization for perimedullary arteriovenous fistula(PMAVF).Methods Data of 8 PMAVF patients who underwent endovascular embolization were retrospectively analyzed.Findings of digital subtraction angiography(DSA)immediately and 6 months after treatment were reviewed,and the modified Aminoff-Logue score(mALS)was compared before and 6 months after treatment.Results Among 8 cases of PMAVF,there were 2 cases of type Ⅰ,5 cases of type Ⅱand 1 case of type Ⅲ,among them 3 were treated with Onyx glue while 5 with Glubrane glue.The immediate DSA results after treatment showed complete embolization of the fistula in all 8 cases,while those 6 months after treatment showed no recurrence.One case recovered and the other 7 cases improved,and mALS(2.25±2.12)6 months after treatment was lower than that(5.50±2.39)before treatment(P＜0.05).Conclusion Endovascular embolization could safely and effectively treat PMAVF.

OBJECTIVE：To study the prot ective effects of crocin （CR）against triptolide （TP）-induced visceral organ injury in mice，and to provide reference for the studying TP compatibility and detoxification. METHODS ：Fifty mice were randomly divided into normal group ，TP low-dose and high-dose groups （i.e. TP-L group ，TP-H group ，with 300，600 μg/kg），TP low-dose and high dose combined with CR groups （i.e. TP-L+CR group ，TP-H+CR group ，with 300 μg/kg TP+100 mg/kg CR ，600 μg/kg TP+ 100 mg/kg CR ），with 10 mice in each group. Except for normal group ，other groups were given relevant medicine intragastrically ， once a day ，for consecutive 7 d. The body weight of mice was weighted every day ，and their death was recorded. After last administration，the mice were sacrificed ，and the heart ，liver，kidney and testis were taken ，and the organ index was calculated ； serum levels of ALT ，AST，BUN and Scr ，the activity of T-SOD and the contents of MDA were all determined. The pathological changes of heart ，liver，kidney and testis were observed ；mRNA expression of Bcl- 2，Bax and caspase- 3 in liver tissue were determined. RESULTS ：Three，five，two and three mice in TP-L group ，TP-H group ，TP-L+CR group and TP-H+CR group died respectively，and the survival rates were 70％，50％，80％ and 70％，respectively. Compared with normal group ，the body weight （7th day of experiment ），heart index ，liver index ，kidney index （except for TP-L group ），testicular index ，T-SOD activity and mRNA expression of Bcl- 2 in liver tissue ，serum levels of ALT （except for TP-L group ），AST（except for TP-L group ），BUN and Scr，MDA content and mRNA expression of Bax ，mRNA expression of caspase- 3 in liver tissue were increased significantly （P＜ 0.05 or P＜0.01）. There were obvious pathological changes in heart ，liver，kidney and testis tissue. Compared with the same dose of TP alone group ，the above indexes of TP combined with CR group were improved in varying degrees. Except for the renal index and serum ALT level of TP-L+CR group ，there was statistical significance for all indexes （P＜0.05 or P＜0.01）；the pathological injuries of heart ，liver，kidney and testis were significantly improved. CONCLUSIONS ：CR can relieve the damage of heart ， liver，kidney and testis induced by TP ，which may be related to the antioxidant stress of CR.

OBJECTIVE To develop a whole-process intelligent model of pharmaceutical care for peritoneal dialysis （PD） patients， and to provide a reference for clinical pharmacists to provide standardized PD pharmaceutical care. METHODS The pharmaceutical care mode of PD patients at home and abroad was investigated and analyzed. Based on the actual situation of the First Affiliated Hospital of Soochow University （hereinafter referred to as “our hospital”）， with “home→PD center outpatient→ inpatient department” as the main node， the recycling process of medication reconciliation was optimized. The whole-process intelligent pharmaceutical care model of PD was illustrated by improving the Chinese version of the drug-related problems （DRPs） classification tool， developing the corresponding pharmaceutical care process， and presenting specific cases. RESULTS Based on the medication therapy management （MTM） platform， our hospital had built a closed-loop PD whole-process intelligent pharmaceutical care model of “in-hospital pharmaceutical care （building document）-PD outpatient MTM-home pharmaceutical care （online App management）”. A “double cycle” workflow of “admission→discharge→outpatient” medication reconciliation cycle and “discovery-analysis-intervention-follow-up-record-evaluation” DRPs cycle was formed. CONCLUSIONS The establishment of the whole-process intelligent pharmaceutical care model for PD in our hospital provides experience for standardizing pharmaceutical care for PD patients， and can reduce DRPs.