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 evaluate bacteriophage therapy for gut-derived sepsis caused by Acinetobacter baumannii in mice.Methods Lyric phages of Acinetobacter baumannii were isolated from environment by using double-layer agar method.A murine model of gut-derived sepsis was established by oral administration of ampicillin-resistant Acinetobacter baumannii and injection of ampicillin and cyclophosphamide into peritoneal cavities of mice.Bacteriophage therapy were given 1 d before infection (group 1),2 d(group 2) and 6 d(group 3) after infection.The survival of the mice was observed,mice without bacteriophage therapy were as control.Independent-sample t test was performed to compare inflammatory cytokines levels in peripheral blood and liver,number of bacteria in liver and spleen between mice with and without bacteriophage therapy.Results The minimal lethal dose of Acinetobacter baumannii for mice with gut-derived sepsis was 1 × 107 CFU/mL.The survival of the mice in group 2 (4/6 survived),which were treated with bacteriophage 2 d after inoculation of Acinetobacter baumannii,was higher than those of group 1 (2/6 survived),group 3 (3/6 survived) and the control group (phage-untreated,0/6 survived).Interleukin (IL)-1 β,tumor necrosis factor (TNF)-α and IL-6 in peripheral blood in mice with bacteriophage therapy were (105 ±6) ng/L,(105 ± 11) ng/L and (104 ± 12)ng/L,which were lower than those in control group (t =5.04,9.05 and 9.33,P ＜ 0.01) ; IL-1 β and TNF-α in liver of mice with bacteriophage therapy were (104 ± 9) ng/L and (104 ± 11) ng/L,which were lower than those in the controls (t =13.70 and 12.80,P ＜0.01),but IL-6 levels were not of statistical difference between therapy and control groups (t =1.06,P ＞ 0.05).Number of bacteria in liver and spleen in mice with bacteriophage therapy were (2.9 ± 1.3) × 103CFU/g and (8.3 ±7.6) × 102 CFU/g,which were also lower than those in control group (t =9.16 and 8.96,P ＜ 0.01).Conclusions Bacteriophage therapy can be effective against gut-derived sepsis caused by Acinetobacter baumannii.

ObjectiveTo investigate the association of high-density lipoprotein cholesterol (HDL-C) with the prognosis of patients with alcohol-related hepatocellular carcinoma (HCC) after radical treatment. MethodsA retrospective analysis was performed for the clinical data of 43 patients with alcohol-related HCC who were admitted to The Fifth Medical Center of Chinese PLA General Hospital and underwent radical treatment from January 2008 to July 2015, and according to HDL-C level, the patients were divided into normal group with 26 patients and abnormal group with 17 patients. The two groups were compared in terms of basic information, laboratory markers, imaging indices, Barcelona Clinic Liver Cancer tumor stage, and Child-Pugh class of liver function. The t-test test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. The Kaplan-Meier method was used to plot survival curves and the log-rank test was used for comparison between groups. Univariate and multivariate Cox proportional hazards models were used to analyze independent risk factors for prognosis. ResultsThere was a significant difference in prealbumin between the two groups (162.38±60.86 mg/L vs 120.06±64.08 mg/L, t=2.184, P=0.035). Number of tumors (hazard ratio ［HR］=2.839, 95%confidence interval ［CI］: 1.120~7.200,P=0.028), tumor size (HR=2.634, 95%CI: 1.062~6.529,P=0037), and HDL-C level (HR=2.400, 95%CI: 1.040~5.537,P=0.040) were independent risk factors for the overall survival of patients with alcohol-related HCC. There were significant differences in 1-, 3-, and 5-year cumulative survival rates between the normal group and the abnormal group (88.5%/72.4%/55.7% vs 70.6%/43.7%/17.5%, χ2=5.881, P=0.015). ConclusionThe reduction in HDL-C level might indicate poor prognosis of patients with alcohol-related HCC.

Objective:To investigate the role of TRIZ theory in clinical teaching in orthopedic operating room.Methods:A total of 43 interns in the orthopedic operating room of our hospital from October 2019 to July 2020 were selected as control group, and 46 interns in the orthopedic operating room of our hospital from August 2020 to May 2021 were selected as observation group. The interns in the control group received traditional teaching, and those in the observation group received teaching based on TRIZ theory. The two groups were compared in terms of professional assessment results, self-evaluation, satisfaction score, and qualification of aseptic operation in orthopedic operating room. SPSS 22.0 was used to perform the t-test and the chi-square test. Results:Compared with the control group, the observation group had significantly higher theoretical knowledge score, practical skill score, and weighted total score ( t=14.90, 11.82, and 13.23, all P?0.001). Compared with the control group, the observation group also had significantly higher scores of analysis and problem-solving ability, clinical thinking, doctor-patient communication, psychological quality, clinical case analysis, self-study ability, and teaching satisfaction ( t=7.88, 11.78, 15.26, 7.94, 12.93, 9.83, and 13.22, all P?0.001). Compared with the control group, the observation group had significantly higher qualified rates of surgical hand-washing, wearing sterile surgical gowns, non-contact glove wearing; and bacteriological testing ( χ 2=4.22, 6.02, 4.92, and 7.59, P=0.040, 0.014, 0.027, and 0.006). Conclusion:TRIZ theory has a certain value in clinical teaching in orthopedic operating room and can significantly improve the scores of theoretical knowledge and practical skills, strengthen aseptic operation abilities in orthopedic operating room, and enhance the satisfaction with teaching among interns.

Background and objective Paclitaxol (PTX) resistance is one of main factors which affect the outcome of chemotherapy of lung adenocarcinoma. The aim of this study is to compare the secreted protein expression profiles between Paditaxol (PTX) resistant and sensitive lung adenocarcinoma cells by proteomic research method, so as to provide evidence of choosing individual chemotherapy drugs in clinical treatment. Methods Total secreted proteins extracted from a PTX sensitive cell line A549 and a PTX resistant cell line A549-Taxol were separated by fluorscent differential gel electrophoresis (DIGE). High quality 2-DE profiles were obtained and analyzed by Decyder 6.5 analysis software to screen differentially expressed protein spots. Those spots were identified by mass spectrometry. Results 2-DE patterns of lung adenocarcinoma cells with high-reso-lution and reproducibility were obtained. 76 significantly differentially expressed protein spots were screened, 19 proteins were identified by mass spectrometry. The identified proteins could be classified into different catogories: metabolic enzyme, extracel-lular matrix (ECM) degradation enzyme, cytokine, signal transducer, cell adhesion, and so on. Conclusion Multiple secreted proteins related to chemoresistance of A549-Taxol cells were identified in this study for the first time. The results presented here would provide dues to identify new serologic chemoresistant biomarkers of NSCLC.

Objective:To review our clinical experience on total pancreatectomy in treatment of pancreatic diseases.Methods:The clinical data of 25 patients with pancreatic diseases who underwent total pancreatectomy at Nanjing Drum Tower Hospital from February 2016 to October 2019 were retrospectively analyzed. The data on general information, surgical methods, postoperative complications, changes in postoperative pancreatic endocrine and exocrine functions and quality of life were analyzed.Results:There were 16 males and 9 females, aged (60.9±9.4) years. Planned selective surgery was carried out in 16 patients while unplanned operations in 9 patients. There were 15 patients with pancreatic neoplasms with 4 patients who received neoadjuvant or conversion therapy, 6 patients with intraductal papillary mucinous neoplasms, 3 patients with chronic pancreatitis and 1 patient with pancreatic neuroendocrine carcinoma. The total postoperative complication rate was 36% (9/25). One patient died in the perioperative period, and one patient underwent a second operation. After 6 months of operation, 2 of 24 patients had died. The remaining patients had a fasting blood sugar of (8.9±1.6) mmol/L, with an insulin dosage of (30.7±10.6) U/d. The average dosage of trypsin was (1.1±0.3) g/d. The quality of life score reached or exceeded the preoperative levels.Conclusions:Total pancreatectomy was safe and feasible for some pancreatic diseases. For patients with pancreatic cancer, its long-term oncological outcomes need further studies.

Objective:To evaluate the diagnostic performance of non-contrast-enhanced Dixon water-fat separation Compressed SENSE (CS-SENSE) whole-heart coronary magnetic resonance angiography (CMRA) at 3.0 T on patients with suspected coronary artery disease (CAD).Method:The study complied with the Declaration of Helsinki. Local ethics committee approved this study and written informed consent was obtained from each patient. In this prospective study, from March 2021 to September 2021, 53 consecutive participants with suspected CAD who were scheduled for X-ray coronary angiography (CAG) were prospectively recruited in Zhongshan Hospital. CMRA was performed with a 3.0 T scanner without contrast agent enhancement during free breathing with Dixon water-fat separation and CS-SENSE methods. The accuracy of CMRA for detecting a ≥ 50% reduction in diameter was determined using CAG as the reference method.Results:Acquisition of whole-heart CMRA images was successfully performed in 46 (86.8%) of 53 patients with an average imaging time of (7.8±1.8) min. The sensitivity, specificity, positive predictive values, negative predictive values, and accuracy of CMRA according to a patient-based analysis were 95.8%(95%CI 78.9%-99.9%), 81.8%(95%CI 59.7%-94.8%), 85.2%(95%CI 66.3%-95.8%), 94.7%(95%CI 74.0%-99.9%), 89.1%(95%CI 76.4%-96.4%), respectively. The areas under the receiver-operator characteristic curve (AUC) from CMRA images according to patient-, vessel-and segment-based analyses were 0.876(95%CI 0.745-0.955), 0.880(95%CI 0.814-0.929), 0.903(95%CI 0.877-0.926), respectively.Conclusion:3.0 T non-contrast-enhanced Dixon water-fat separation CS-SENSE whole-heart CMRA is a promising technique to detect clinically significant coronary stenosis on patients with suspected CAD.

ABSTRACT:Objective To assess the effect of nurse intervention of urea-based cream (UBC)on sorafenib-induced hand-foot skin reaction (HFSR)in patients with advanced hepato-cellular carcinoma(HCC).Methods 202 advanced HCC patients were randomly divided into 2 groups.101 patients in the urea-based cream group were applied with 10% UBC three times dai-ly and best nursing while 110 patients in the control group were applied basic nursing alone.The treatment lasted for 12 weeks.HFSR was assessed every 2 weeks.Once HFSR occurred,patients were allowed any cream,including UBC.Results After the nursing,the incident rate of HFSR and incidence of grade ≥2 HFSR in the UBC group were lower that than in the BNC group.The Median time to first occurrence of HFSR was 1.63 times of UBC group.UBC did not affect the adjustment of sorafenib dose,response or disease control.Conclusion UBC nurse intervention of advanced HCC patients could reduce HFSR rate,extend the time of first occurrence of HFSR and improve the quality of life of patients.

ABSTRACT:Objective To assess the effect of nurse intervention of urea-based cream (UBC)on sorafenib-induced hand-foot skin reaction (HFSR)in patients with advanced hepato-cellular carcinoma(HCC).Methods 202 advanced HCC patients were randomly divided into 2 groups.101 patients in the urea-based cream group were applied with 10% UBC three times dai-ly and best nursing while 110 patients in the control group were applied basic nursing alone.The treatment lasted for 12 weeks.HFSR was assessed every 2 weeks.Once HFSR occurred,patients were allowed any cream,including UBC.Results After the nursing,the incident rate of HFSR and incidence of grade ≥2 HFSR in the UBC group were lower that than in the BNC group.The Median time to first occurrence of HFSR was 1.63 times of UBC group.UBC did not affect the adjustment of sorafenib dose,response or disease control.Conclusion UBC nurse intervention of advanced HCC patients could reduce HFSR rate,extend the time of first occurrence of HFSR and improve the quality of life of patients.

Objective:To analyze the distribution of clopidogrel metabolism-related gene variability in Kawasaki disease (KD) children with coronary artery lesions (CAL) across different age groups and the impact of genetic variability on the efficacy of clopidogrel antiplatelet therapy.Methods:A retrospective cohort study was conducted. Clinical data were collected from 46 KD children with CAL who were hospitalized in the Cardiovascular Center of Children′s Hospital of Fudan University between January 2021 and August 2022 and were treated with clopidogrel, including gender, age, body mass index, course of KD, CAL severity grade, and baseline platelet count. According to their age, the children were divided into ≥2-year-old group and <2-year-old group. Their platelet responsiveness was assessed by adenosine diphosphate-induced platelet inhibition rate (ADPi) calculated via thromboelastography, and children were categorized into high on-treatment platelet reactivity (HTPR) and normal on-treatment platelet reactivity (NTPR) groups. Genotypes of CYP2C19, PON1 and ABCB1 were detected. The t test, one-way analysis of variance and Chi-square test were used for intergroup comparison. Results:Among the 46 KD children with CAL, 34 were male and 12 were female; 37 were ≥2-year-old and 9 were <2-year-old; 25 cases were in the HTPR group and 21 cases were in the NTPR group, with 19 HTPR and 18 NTPR in the ≥2-year-old group, and 6 HTPR and 3 NTPR in the <2-year-old group. Genetic analysis showed that 92 alleles among the 46 children, with frequencies of CYP2C19*1, CYP2C19*2, CYP2C19*3, CYP2C19*17, PON1 192Q, PON1 192R, ABCB1 3435C, ABCB1 3435T at 59% (54/92), 32% (29/92), 9% (8/92), 1% (1/92), 36% (36/92), 64% (59/92), 63% (58/92) and 37% (34/92), respectively. Analysis of the impact of genotype on ADPi revealed that in children aged ≥2 years, those with CYP2C19*1/*3 genotype had significantly lower ADPi than those with CYP2C19*1/*1 genotype ((34±15)% vs. (61±29)%, t=2.18, P=0.036). There were also no significant difference in ADPi among children with PON1 192Q homozygous, PON1 192R heterozygote and PON1 192R homozygous genotypes ((40±22)% vs. (52±33)% vs. (65±27)%, F=2.17, P=0.130), or among those with ABCB1 3435C homozygous, ABCB1 3435T heterozygote and ABCB1 3435T homozygous genotypes ((55±34)% vs. (60±27)% vs. (49±24)%, F=0.33, P=0.719). In <2-year-old group, there were no significant differences in ADPi across CYP2C19*1/*1, CYP2C19*1/*2 and CYP2C19*2*2 genotypes ((40±20)% vs. (53±37)% vs. (34±16)%, F=0.37, P>0.05). There were no significant differences in ADPi across CYP2C19*1/*1 and CYP2C19*1/*3 genotypes ((44±27)% vs. (42±20)%, t=0.08, P>0.05). There were no significant differences in ADPi across PON1 192Q homozygous, PON1 192R heterozygote and PON1 192R homozygous genotypes (45% vs. (55±27)% vs. (24±5)%, F=1.83, P>0.05). There were no significant differences in ADPi across ABCB1 3435C homozygous, ABCB1 3435T heterozygote and ABCB1 3435T homozygous genotypes ((36±16)% vs. (50±35)% vs. 45%, F=0.29, P>0.05). The risk analysis of HTPR in different genotypes revealed that in children aged ≥2 years, carrying at least 1 or 2 loss-of-function alleles of CYP2C19 was a risk factor for HTPR ( OR=4.69, 10.00, 95% CI 1.11-19.83, 0.84-119.32, P=0.033, 0.046, respectively), and PON1 192R homozygosity and carrying at least one PON1 192R allele were protective factors against HTPR ( OR=0.08, 0.13, 95% CI 0.01-0.86, 0.01-1.19, P=0.019, 0.043, respectively). Conclusion:KD children aged ≥2 years carrying CYP2C19 loss-of-function alleles and PON1 192Q are more likely to develop HTPR.