Objective To investigate the clinical significance of PRL-3 expression in sinonasal squamous cell cancer(SNSCC).Meth-ods The immunohistochemical method and RT-PCR were adopted to detect the PRL-3 protein expression level in 62 cases of SNSCC tissue (SNSCC group),30 cases of nasal polyps(NP group)tissue and 25 cases of normal nasal mucosa tissue(control group).The obtained results were compared.Results Both in the protein level and gene level detection,the expression of PRL-3 in the SNSCC group was higher than that in the NP group and control group,the difference was statistically significant (P <0.05).The expression of PRL-3 had no significant differences among different ages and between different genders(P >0.05),but with TNM stage increasing,differentiation degree decreasing and complicating lymph node metastasis,the expression of PRL-3 was significantly increased(P <0.05).Conclusion The PRL-3 expression can serve as good reference for the proliferation activity of SNSCC,its expressing intensity can obviously reflect the SNSCC cell proliferation activity,PRL-3 probably is an independent prognostic index of SNSCC,indicating poor prognosis.

Objective To identify the drug resistance-related genes in a clinically isolated strain of Enterobacter cloacae.Methods A strain of Enterobacter cloacae was isolated from sputum of a patient with chronic obstructive pulmonary disease from the Affiliated Hospital of Xuzhou Medical University in March 2013.Modified Hodge test and metal enzyme inhibition test were performed for drug-resistant phenotype screening.Carbapenemase genes blaMUS-1, blaVIM-1, blaVIM-2, blaIMP, blaKPC-2, blaNDM-1, blaOXA-48 and blaGESwere amplified by polymerase chain reaction (PCR), and the positive products were sequenced and analyzed.Plasmid conjugation and transformation experiments were used to confirm that the resistance gene mediated by plasmids.Agar dilution method was used for antibiotic susceptibility test.Results Both modified Hodge test and metal enzyme inhibition test were positive in this strain of Enterobacter cloacae.blaNDM-1 gene and blaKPC-2 gene were detected by PCR, and further confirmed by sequencing.blaNDM-1 gene was carried by IncX plasmid with 54×103 bp, KPC-2 gene was carried by untyping plasmid with 42×103 bp.The strain was only sensitive to tetracycline (MIC=2 μg/mL) and tigecycline (MIC=1 μg/mL).The symptoms were improved after the patient was treated by tigecycline combined with Piperacillin/Tazobactam.Conclusion blaNDM-1 and blaKPC-2 genes in Enterobacter cloacae can be mediated by plasmids, and appropriate therapy for its infection should be based on the result of antibiotic susceptibility test.

Objective To investigate the mechanism related to reduced antibiotic sensitivity of Acinetobacter baumannii inducted by meropenem in vitro.Methods Three strains of clinically isolated carbapenems-sensitive Acinetobacter baumannii were induced by meropenem in vitro, and the mutant strains (MS1, MS2 and MS3) were obtained.Minimal inhibitory concentrations (MICs) of antimicrobial agents to strains before and after induction were determined by automatic drug sensitivity analyzer .The homology of strains was analyzed by Enterobacterial repetitive intergenic consensus -polymerase chain reaction ( ERIC-PCR).Modified Hodge test and EDTA-Na2-double disk synergy test were used to detect carbapenemase and metallo-β-lactamase (MBL), respectively.Main carbapenemase genes were detected by PCR and followed by DNA sequencing.Expressions of adeB and outer membrane proteins in strains before and after induction were detected with fluorescence quantitative PCR and SDS -polyacrylamide gel electrophoresis , respectively.t test was used for data analysis .Results The sensitivity of mutant Acinetobacter baumannii strains to meropenem and most antibiotics was reduced , except to imipenem, amikacin and polymyxin; and the reduced sensitivity to meropenem in MS2 and MS3 was of genetic stability.ERIC-PCR showed 100%homology between the mutant strains and parental strains .Both carbapenemase and metallo -β-lactamase were negative in mutant strains and parental strains , and only OXA-51 gene was found.The expressions of adeB gene in mutant strains were 24.26 ±0.91, while those in parental strains were 22.81 ±0.38, and the difference was not significant (t =2.534, P >0.05).Outer membrane protein with molecular weight 54 000 was missing in MS1, while that with molecular weight 47 000 was missing in MS2 and MS3.Conclusion Reduced antibiotics sensitivity in meropenem -induced Acinetobacter baumannii may be correlated with the deficiency of outer membrane protein with molecular weight 47 000.

Objective To explore the optimal conditions of preparing 68Ga-DOTA-iNGR (NGR peptide containing CendR motif),to evaluate its biodistribution in normal mice and to perform microPET imaging in tumor-bearing nude mice.Methods 68Ga fresh eluent (200 μl,92.5-129.5 MBq) obtaining with 68Ge-68Ga radionuclide generator was used to label DOTA-iNGR.The optimal conditions of labeling including pH,temperature,reacting time and concentration of DOTA-iNGR were determined.Then,the in vitro and in vivo stability and octanol/water partition coefficient of 68Ga-DOTA-iNGR were further analyzed.The biodistribution in normal Kunming mice was examined at 10,20,40,60 and 120 min after injection of 68Ga-DOTA-iNGR.Nude mice bearing HT-1080 (CDl3-positive) and HT-29 (CDl3-negative) tumors were established and underwent microPET imaging at 1 h after the intravenous injection of 68Ga-DOTA-iNGR.Data were analyzed using independent-sample t test.Results The optimal conditions of labeling was mixing 2 μg DOTA-iNGR peptide with 200 μl 68Ga (92.5-129.5 MBq) at pH 4.0,temperature 90-100 ℃ for 5-10 min.Under this condition,labeling rate reached (97.5± 1.3)％.The radiochemical purity of 68Ga-DOTA-iNGR in both saline (room temperature) and mouse serum (37 C) were both above 95％ after 4 h incubation,and the radiochemical purity in urine was greater than 85％ after 1 h metabolism in vivo.The partition coefficient was-2.71±0.18.In normal mice,majority of 68Ga-DOTA-iNGR was excreted from kidneys with a rapid clearance from blood.The in vivo microPET imaging showed that 68Ga-DOTA-iNGR was remarkably accumulated in the CD13-positive HT-1080 tumor.Conclusions Labeling DOTA-iNGR with 68Ga under our condition is a simple and efficient procedure with high labeling rate and high specificity.The product 68Ga-DOTA-iNGR has high stability,ideal biodistribution,and specific binding to CD13-positive tumor,which means that it's a very promising molecular probe for noninvasively detecting CD13-positive tumor.

Objective To investigate DNA methylation markers in the whole blood of patients with systemic lupus erythematosus(SLE),in hope to facilitate the evaluation of SLE severity.Methods Whole blood samples were obtained from 58 patients with SLE(including 14 cases of severe SLE,25 moderate SLE,19 inactive SLE)and 50 healthy controls.Bisulphite sequencing was performed to determine the methylation status of interleukin-2 common receptor gamma chain(IL-2RG)promoter region,and real-time reverse transcriptionPCR to quantify the expression level of IL-2RG mRNA,in these subjects.Results The methylation level of IL2RG promoter region was 0.217 ± 0.140,0.325 ± 0.230,0.342 ± 0.085 and 0.175 ± 0.036 in the patients withsevere,moderate and inactive SLE and healthy controls,respectively.A significant increase was observed in the methylation level of IL-2RG promoter region in the patients with inactive SLE compared with the patients with severe SLE and healthy controls(both P ＜ 0.01),and in the patients with SLE compared with the healthy controls(0.263 ± 0.047 vs.0.175 ± 0.036,P ＜ 0.05).The expression level of IL-2RG mRNA was significantly lower in the patients with SLE than in the healthy controls(2.550 ± 0.823 vs.4.293 ± 1.283,P ＜ 0.05).A negative correlation was observed between the expression level of IL-2RG mRNA and methylation level of IL2RG promoter region in 20 patients with SLE(r =-0.44,P ＜ 0.05).Conclusion The methylation status of IL2RG promoter region is statistically higher in patients with SLE than in healthy controls,and significantly different between patients with active SLE and those with stable SLE.

Objective To quantitatively compare the diagnostic capability of 68Ga-NGR and 18F-FDG in well-differentiated hepatocellular carcinoma (HCC) bearing mice by microPET/CT imaging.Methods The in vitro cellular uptake, in vivo microPET/CT imaging and biodistribution studies of 68Ga-NGR and 18F-FDG were quantitatively compared in SMMC-7721-based well-differentiated HCC.The human fibrosarcoma (HT-1080) and human colorectal adenocarcinoma (HT-29) cells/xenografts were respectively used as positive and negative reference groups for CD13.The expression of CD13 was qualitatively verified by immunohistostaining.The levels of CD13 and glucose-6-phosphatase (G6Pase) were semi-quantitatively analyzed by Western blot test for all 3 types of tumors.Two-sample t test was used for data analysis.Results The in vitro cellular uptake showed that the 68Ga-NGR uptake in SMMC-7721 and HT-1080 cells was higher than that in HT-29 cells, and the 68Ga-NGR uptake was higher than 18F-FDG uptake in SMMC-7721 cells.The in vivo microPET/CT imaging results revealed that the uptake of 68Ga-NGR in SMMC-7721 tumor was (2.17±0.21) %ID/g, remarkably higher compared to (0.73±0.26) %ID/g of 18F-FDG uptake (t=8.826, P<0.01).The tumor/liver ratio of 68Ga-NGR was 2.05±0.16, which was 2.03-fold higher than that of 18F-FDG.In the HT-1080 tumors, the uptakes of 68Ga-NGR and 18F-FDG were both high, and the values were (2.46±0.23) %ID/g, (3.47±0.31) %ID/g.The uptake of 68Ga-NGR was significantly lower than that of 18F-FDG in HT-29 tumors: (0.67±0.20) %ID/g vs (3.17±0.29) %ID/g;t=4.221, P<0.01.Western blot and immunohistostaining results were as follows: HT-1080(CD13+, G6Pase-), SMMC-7721(CD13+, G6Pase+), HT-29(CD13-, G6Pase-).Conclusions The uptake of 68Ga-NGR is higher than 18F-FDG uptake in SMMC-7721 tumor bearing mice, therefore it is worthwhile to consider the feasibility of clinical translation for PET/CT in diagnosis of HCC.Furthermore, because of the difference in 68Ga-NGR and 18F-FDG avidities in tumors with different molecular phenotypes of CD13 and G6Pase, there is an underlying potential for molecular imaging in the determination of molecular phenotypes.

OBJECTIVETo evaluate the efficacy and adverse effects of weekly irinotecan combined with capecitabine as a second-line chemotherapy for treatment of advanced gastric cancer.

METHODSTwenty-one patients with advanced gastric cancer who had failed first-line therapy received irinotecan on days 1 and 8 plus capecitabine on days 1-14 for a 21-day cycle. Each patient was treated for at least two cycles and evaluated 4 weeks later for the responses.

RESULTSOf the 21 patients, none showed complete remission (CR), 5 (23.8%) showed partial remission (PR), 6 (28.6%) showed stable disease (SD) and 10 (47.6%) showed progressive disease (PD). The overall response rate was 23.8%, and 11 patients (52.4%) benefited (CR+PR+SD) from the clinical therapy, with a mean time to tumor progression of 3.61±0.97 months. The main adverse effects of this regimen included myelosuppression, nausea, vomiting and diarrhea.

CONCLUSIONThe regimen of weekly irinotecan plus capecitabine has a definite effect for treatment of advanced gastric cancer with tolerable toxicity.

Adenocarcinoma ; drug therapy ; pathology ; Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Camptothecin ; administration & dosage ; analogs & derivatives ; Capecitabine ; Deoxycytidine ; administration & dosage ; analogs & derivatives ; Female ; Fluorouracil ; administration & dosage ; analogs & derivatives ; Humans ; Male ; Middle Aged ; Stomach Neoplasms ; drug therapy ; pathology ; Treatment Outcome

Objective To understand the prevalence of resistance gene and homology of carbapenem-resistant Klebsiella pneumoniae (CRKP) isolated from ICU of emergency.Methods A total of 19 CRKP isolates were obtained from emergency ICU of the Affiliated Hospital of Xuzhou Medical University from July 2015 to August 2016.PCR was performed to screen the genes encoding carbapenemase,extended spectrum beta-lactamase (ESBL) and AmpC.Pulsed field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST) were used for molecular typing of these bacterial strains.Results Among the 19 CRKP,carbapenemase-resistant genes were detectable in 18 CRKP isolates,including 17 isolates of harboring blaKPC gene and 1 strain of harboring blaNDM gene.All the 18 strains carried ESBLs genes which were identified as 8 blaSHV-12,3 blaSHV-11,5 blaSHV-2a,15 blaTEM-1,10 blaCTX-M-65,3 blaCTX-M-15,1 blaCTX-M-14 and 1 blaCTX-M-27.The 13 strains harboring cephalosporin-resistant genes were all identified as blaDHA-1.PFGE results revealed that the 19 CRKP strains were grouped into 4 types (A,B,C and D) and 4 subtypes(A1,2,3 and 4):A1 (n =12),A2(n =1),A3 (n=1),A4(n=1),B(n=2),C(n=1) and D(n=1).MLST showed that ST11 was the predominant sequence type (n=15) among the 19 CRKP strains,and ST48 (n =2),ST37 (n =1) and untyped (n =1) were also identified.The 15 blaKPC-2-producing CRKP ST11 clone shared the A type of PFGE pattern.Conclusion The report on CRKP suggested the dissemination of blaKPC-2-producing ST11 clone was existed in the ICU of emergency department in this hospital.The surveillance for drug-resistance and effective disinfectant quarantine measures should be strengthened.

Objective: To investigate the effect of moxibustion at Shenque (CV 8) on fatigue in rats with chronic exercise-induced exhaustion. Methods: Thirty male Sprague-Dawley (SD) rats were randomly divided into a blank group, a model group and a moxibustion group, 10 rats in each group. Except rats in the blank group, the remaining rats were subjected to create long-term exhaustion models by repeated swimming. After successful modeling, rats in the moxibustion group received mild moxibustion at Shenque (CV 8) for 15 min, once every other day with a total of 10 times. Rats in the model group and the blank group did not receive moxibustion. At the end of the treatment, the exhausted times, and the body weight of rats before and after the experiment were compared among groups. The levels of blood malondialdehyde (MDA) and urea nitrogen (BUN), as well as the activities of aspartate transarninase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) were also measured by the automatic biochemical analyzer, 24 h after the exhausting excise. Results: The 10th swimming time was significantly longer in the moxibustion group than that in the model group (P<0.01). The increase rate of the body weight was lower in the rats of the moxibustion group than that in the model group before the 7th and the 10th exhausting excise (P<0.05, P<0.01). The levels of serum MDA and BUN, as well as the activities of AST, ALT and LDH in the model group were higher than those in the blank group (all P<0.01). The levels of serum MDA and BUN, as well as the activities of AST, ALT and LDH in the moxibustion group were lower than those in the model group (P<0.01). Conclusion: Moxibustion at Shenque (CV 8) can decrease the serum levels of MDA and BUN, as well as activities of AST, ALT and LDH in the long-term fatigue rats, thus to improve the symptoms of fatigue.

Objective To report our experience with retroperitoneoscopic nephroureterectomy with excision of a bladder-cuff for renal pelvic and ureteral tumors.Methods Thirty-five patients (21 women and 14 men;mean age,67 years;age range,49 -82 years) with upper urinary tract tumors underwent retro- peritoneal laparoscopic nephrourcterectomy with excision of a bladder-cuff.Of the 35 cases,15 had pelvic tumors and 20 had ureteral tumors;19 cases had the tumors on the right side and 16 on the left.Two cases had ureteral tumors combined with bladder tumors.One case had bilateral ureteral tumors then concomitantly had bladder tumors.The needle electrode was used to circleround incise the bladder thoroughly 0.5 cm away from the ureterostoma.Three trocars in the waist were used for dissecting the kidney;and the ureter was dis- sected as far distally downward.Then an incision of 5-9 cm was created in the lower abdomen to allow dis- section of the distal ureter and bladder-cuff and intact specimen extraction.Results The operation was successful in all 35 patients.The mean operative time was 3.1 h ( range,1.5-6.0h).The mean estimated blood loss was 166 ml (range,20-1600 ml).Four cases received blood transfusion.The patient's activity re- covered in 20-32 h after operation.Postoperative pathology showed transitional cell carcinoma in 30 cases, poorly differentiated adenocareinoma in 2 (ureter),squamous cell carcinoma in 1 (ureter),leiomyosarcoma in 1 (ureter),xanthogranulomatous pyelonephritis in 1.Duodenal leakage occurred in 1 patient who had had dialysis a drainage catheter was placed on the third d after operation,and the patient died of heart failure af- ter 2 months.Postoperative vesical irrigation was performed to prevent tumor recurrence.The mean hospital stay was 11 d.During a mean follow-up of 14 months(range,1-32 months),1 patient developed pelvic me- tastasis and was alive with the tumor.The other 33 patients survived free of tumor to date.No patient had re- current transitional cell carcinoma of the bladder.Conclusions Our data demonstrate that retroperitoneo- scopic nephroureterectomy for renal pelvic and ureteral tumors has shorter incision and more rapid postopera- tive recovery compared with open surgery.Using resectoscope to resect the termination of ureter allows more complete excision of the ureter.