[Use of alternative pancreatic fistula risk score system for patients with clinical relevant postoperative pancreatic fistula after laparoscopic pancreaticoduodenectomy].

Objective: To examine the application value of alternative pancreatic fistula risk score system(a-FRS) for patients with clinically relevant postoperative pancreatic fistula(CR-POPF) after laparoscopic pancreaticoduodenectomy(LPD). Methods: Clinical data of 400 patients who underwent LPD at Department of Hepatobiliary and Pancreatic Surgery,Jilin University First Hospital,from April 2015 to August 2019 were retrospectively analyzed.There were 217 males and 183 females, with age of (M(QR)) 58 (53) years (range:26 to 93 years) and body mass index of (23.0±2.7) kg/m2 (range:19.4 to 27.1 kg/m2).Preoperative CA19-9 was (171.6±212.7) U/ml (range:32.1 to 762.6 U/ml), and preoperative CA125 was (18.6±22.9) U/ml (range:9.0 to 112.3 U/ml).Univariate analysis and multivariate Logistic regression analysis were implemented to find independent risk factors in CR-POPF.According to 3 indicators of a-FRS system(pancreatic texture,main pancreatic duct diameter,and body mass index),receiver operator characteristic curve was used to prospectively analyze the clinical value of CR-POPF. Results: CR-POPF occurred in 60 patients(15.0%) among the 400 LPD patients,including 54 patients(13.5%) with grade B pancreatic fistula and 6 patients(1.5%) with grade C pancreatic fistula.Univariate and multivariate Logistic regression analysis results showed that soft pancreas,diameter of main pancreatic duct ≤3 mm,and body mass index>23 kg/m2 were the independent risk factors for CR-POPF after LPD.The incidence of CR-POPF was 1.9% in the group with low pancreatic fistula risk(0 to 5%),5.9% with moderate pancreatic fistula risk(>5% to 20%),and 80.7% with high pancreatic fistula risk(>20%).a-FRS prospectively predicted the sensitivity and specificity of CR-POPF after LPD was 76.7% and 96.8%,positive predictive value was 80.7%,negative predictive value was 95.9%,positive likelihood ratio was 23.66,negative likelihood ratio was 0.24,and area under the curve was 0.735(95%CI:0.668-0.799). Conclusion: a-FRS system has great clinical application value in predicting CR-POPF after LPD,which can provide basis for early risk prediction of CR-POPF and timely related clinical intervention.

[Relevant molecular characteristics analysis on malignant transformation of interstitial cells induced by tumor stem cells in glioma microenvironment].

Objective: A variety of interstitial cells in tumor microenvironment (TME) based on glioma stem cells(GSC) have the function to promote malignant progression of tumors, but whether these interstitial cells have already undergone malignant transformation and their related molecular characteristics are still poorly understood. Methods: Human SU3-RFP glioma stem cells (GSC) stably transfected with red fluorescent protein (RFP) and interstitial cells from enhanced green fluorescent protein (EGFP) transgenic nude mice were co-cultured in vitro. SU3-RFP cells were also inoculated in different tissues of EGFP-Balb/c nude mice. Immortal EGFP(+) cells were monocloned either from co-culture cells in vitro, or from their xenografts in vivo. These immortal EGFP(+) cells were confirmed to bear characteristics of tumor cell via chromosomal analysis and tumorigenicity assay. Related molecular phenotypes of these cells were further detected through RT-PCR, flow cytometry and immunochemistry(IHC) techniques. Results: (1) Two EGFP(+) cell lines were obtained in vitro, and 5 EGFP(+) cell lines were obtained in vivo tumorigenic experiments. Seven EGFP(+) cell lines all have characteristics of self-renewal, heteroploid of chromosomes and 100% tumorigenicity. (2) Cell surface marker analysis showed cell origin of these cell lines were macrophages (tMΦ1 and tMΦ2 ), dendritic cells (tDC1 and tDC2), fibroblasts (tFB), oligodendrocytes (tOG) and BMSC cells (tBMSC), respectively. (3)All of these seven cell lines co-expressed Sca-1 and c-myc, and have Sox-2 or Nanog expression also, which suggest that they may bear molecular characteristics of mesenchymal stem cells or pluripotent stem cells. Conclusions: (1) Tumor stromal cells in TME have undergone malignant transformation, which is related to the tissue remodeling of TME by GSCs, and not limit to the specific type of their parasitic tissues. (2) Tumor cells originated from GSC and tumor interstitial cells, respectively, are two major types of tumor cells with different origins in glioma parenchyma, can not be simply regarded as tumor heterogeneity, transformed interstitial cells of TME may have the potential to serve as new targets for target diagnosis and therapy.

[Laparoscopic pancreaticoduodenectomy with a novel artery first and uncinate process first approach through Treitz ligament].

Objective: To explore the clinical effect of a novel artery first and uncinate process first approach for laparoscopic pancreaticoduodenectomy(LPD), emphasizing the left lateral and posterior dissection of uncinate process (UP) via Treitz ligament approach. Methods: From April to November 2016, 18 patients received LPD with a novel approach in Pancreas Center of the First Affiliated Hospital with Nanjing Medical University. All patients were diagnosed as pancreatic head or peri-ampulla tumor, without major vessel invasion nor distant metastasis. For resection, routine caudal view was used in the first step, to dissect the anterior medial border between uncinate process and superior mesenteric vein(SMV). Lymphatic tissues were completely dissected form anterior surface of hepatoduodenal ligament. In the second step, left lateral view with camera from left para-umbilical trocar was used, Treitz ligament was incised, SMA root was exposed. After anticlockwise rotation and retraction of mesentery, the anatomic relationship between SMA trunk, inferior pancreaticoduodenal artery(IPDA), jejunal branch of SMV, and distal part of UP, could be perfectly exposed from left lateral view. SMA was dissected from its root until the position above the uncinate process and duodenum, IPDA was transected, distal part of UP was freed from SMA. In the third step, right lateral view and caudal view were alternatively used; proximal UP mesentery was completely dissected out from SMA root, CA root and posterior surface of hepatoduodenal ligament. Pancreaticoduodenectomy was completed in the forth step after transection of pancreatic neck and common hepatic duct. Results: The SMA root and distal UP were successfully dissected out via Treitz ligament approach in all 18 patients, among them, distal UP was completely excised in 8 patients from left view. Postoperative pathology showed R0 resection rate in 69%. Postoperative complication included intra-abdominal hemorrhage in 1 patient, pancreatic fistula in 7 patients(6 cases with grade A and 1 case with grade B), delayed gastric emptying in 4 patients (2 cases with grade A, 2 cases with grade B). Average postoperative hospital stay was (15.5±6.8)days. Conclusion: The novel artery first and uncinate process first approach through Treitz ligament could help surgeons to completely dissect the full length of meso-pancreas along celiac axis-SMA axis in LPD.

Factors associated with Staphylococcus aureus nasal carriage and molecular characteristics among the general population at a Medical College Campus in Guangzhou, South China.

BACKGROUND: The nasal cavity is the main colonization site of Staphylococcus aureus (S. aureus) in human body. Nasal carriage may be a strong risk factor for some serious infection. There was still limited information about the nasal carriage for S. aureus in south China. METHODS: Sought to determine the prevalence and molecular characteristics of S. aureus nasal carriage, 295 volunteers residing on a medicine campus were investigated and sampled the nasal cavity swab. Selected S. aureus isolates were carried through molecular analysis, including pulsed-field gel electrophoresis (PFGE), multilocus sequence analysis, staphylococcal cassette chromosome mec (SCCmec) and virulence gene detection. RESULTS: A total of 73 S. aureus isolates were recovered from separate subjects (24.7%, 73/295), with one methicillin-resistant S. aureus (MRSA) isolate (0.3%, 1/295). Among the 73 isolates, 71 isolates were successfully grouped into 13 pulsotypes by PFGE analysis, with profiles A and L the most prevalent; 12 sequence types (STs) were found among the 23 isolates which had similar drug resistant spectrum. ST59, ST188 and ST1 were the most prevalent, accounting for 17.4, 13.0 and 13.0% of all isolates, respectively. The MRSA isolate presented ST8-SCCmec III. 56.5% of isolates carried both the staphylococcal enterotoxin A (sea) and enterotoxin B (seb) genes. 83.6% of the S. aureus isolates were resistant to penicillin, all isolates were susceptible to quinupristin/dalfopristin, levofloxacin, teicoplanin and vancomycin. The most common risk factors for S. aureus carriage were being male, age ≤30 years, and nasal cavity cleaning habits. CONCLUSIONS: Colonization by S. aureus was greater among male and young age (20-30 years) students and those with irregularity nasal cleaning. The S. aureus isolates selected were revealed into various sequence types and pulsotypes, indicating molecular heterogeneity among S. aureus isolates from the populations in the medical college in Guangzhou.

[Down-regulation of miR-146b-5p promotes malignant transformation of fusion cells after co-culture of macrophages with glioma stem cells in vitro].

Objective: To observe mutual interactions between macrophages(Mφ) and glioma stem cells (GSCs)in dual-color tracing model in vitro, to identify the biological characteristics of fusion cells in multiple levels, and to analysis the relevant molecular mechanisms. Methods: Red fluorescent protein(RFP) gene was stably transfected into human GSCs cell line SU4. Mφ cells were obtained from Balb/c nude mice with enhanced green fluorescent protein (EGFP) expression. Then two cells were co-cultured in dual-color tracing platform. RFP/EGFP double positive cells with high proliferation ability were mono-cloned. The fusion cells were verified by Western blot, fluorescence in situ hybridization, immunocytochemistry and chromosome karyotype analysis.The biological characteristics of fusion cells were further analyzed, together with relevant molecular changes. Results: RFP / EGFP double positive cells were obtained through in vitro co-culture. RFP and EGFP coexpression were proved at transcriptional and translational levels in the fusion cells. They also co-expressed GSCs marker Nestin and Mφ marker CD68, and karyotype analysis showed two types of characteristic chromosomes, which confirmed that the fusion cells originated from spontaneous fusion between SU4-RFP and Mφ.Fusion cell proliferation rate and invasion ability were higher than SU4-RFP, which were relevant with down-regulation of miR-146b-5p and activation of STAT3. Fusion cells transfected with miR-146b-5p showed a higher apoptosis rate(18.83%) and lower tumor formation(4/5). Conclusion: Mφ could fuse with GSCs spontaneously in local tumor micro-environment. The proliferation and invasion abilities of fusion cells were higher than their parent cells, which were relevant with down-regulation of miR-146b-5p and activation of STAT3. It revealed the possible mechanisms of malignant progression of gliomas.

Cloning, expression, and characterization of Fe-SOD from Isöetes sinensis.

Although the palynology and sporophyte stage of Isöetes sinensis have been well studied, the biology of its gametophyte and embryo is less well understood. To date, the functions of several genes of I. sinensis and the molecular mechanisms of enzymes encoded by them remain to be studied. In the present study, the Fe-SOD gene of I. sinensis was successfully cloned using RT-PCR and rapid amplification of cDNA ends (RACE), and termed IsFeSOD. IsFeSOD has certain reference value in the classification of system evolution. The study also accumulated data for further research on the SOD gene. Bioinformatic analysis was employed to compare IsFeSOD with gene sequences obtained from other plants present in the GenBank. Furthermore, the recombinant pET32-FeSOD plasmids were transformed into Escherichia coli BL21 for expression. IsFeSOD was observed to have 1469 nucleotides that were predicted to encode 247 amino acids. The bioinformatic analysis revealed that IsFeSOD contained conserved TGGGA sequences, similar to eight other species, in addition to five other conserved sequences. The recombinant protein was about 43 kDa. Recombinant FeSOD was expressed, purified, and confirmed by western blotting. Alignment of complete Fe-SOD mRNA sequences from 9 species revealed several conserved sequences. A phylogenetic tree was constructed using MEGA4.1 and ClustalX multiple-sequence alignment programs. This study could be helpful in further characterization of SOD genes and for classification of system evolution status.

Analysis of the drug-resistant characteristics of Klebsiella pneumoniae isolated from the respiratory tract and CTX-M ESBL genes.

The main aim of this study was to understand the relationship between the drug-resistant characteristics of Klebsiella pneumoniae and CTX-M-type extended spectrum ß-lactamases (ESBLs), and to detect the distributions of CTX-M-type ESBLs in clinically isolated strains. CTX-M ESBL genes isolated from the clinical samples were amplified by polymerase chain reaction and identified by sequence analysis; the antibiotic susceptibility of the samples was determined using the Kirby-Bauer disc-diffusion method. One hundred and five strains among the 246 isolated strains of K. pneumoniae tested positive for ESBL production (42.68%); 92 of these produced CTX-M ESBLs. Of the 92 CTX-M ESBL strains, 81 produced CTX-M-1 ESBLs and 11 produced CTX-M-25 ESBLs. Fifty-seven of the CTX-M-1 ESBL- and six of the CTX-M-25 ESBL-producing bacteria had CTX-M ESBL genes that coexisted in the plasmid and chromosome. The Kirby-Bauer antibiotic susceptibility method revealed that CTX-M ESBL-positive strains showed a higher rate of resistance to cefazolin, cefoxitin, cefuroxime, ceftazidime, cefotaxime, aztreonam, levofloxacin, and cotrimoxazole, compared to the CTX-M ESBL-negative strains (P < 0.05). The CTX-M ESBL genes were commonly observed in the K. pneumoniae isolated from respiratory tract samples; these were significantly associated with the drug-resistant characteristics of K. pneumoniae to ß-lactam antibiotics.

Correlated expression of Fas, NF-kappaB, and VEGF-C in infiltrating ductal carcinoma of the breast.

OBJECTIVE: To investigate the expressions of Fas, NF-kappaB, and vascular endothelial growth factor-C (VEGF-C) in infiltrating ductal carcinoma of the breast and provide scientific basis for early diagnosis and prognosis of breast cancer. MATERIALS AND METHODS: The immunohistochemical technique (SP method) was used to detect expression of Fas, NF-kappaB, and VEGF-C in 137 cases of breast-infiltrating ductal carcinoma, 17 cases of intraductal carcinoma of the breast, and 20 cases of normal breast tissues, and analyze its relationship with clinicopathologic factors of breast cancer and patients' survival rate, as well as the correlation among their expression, clinicopathologic factors, and survival rate. RESULTS: Fas expression was less commonly detected in infiltrating ductal carcinoma than in intraductal carcinoma and normal tissue. In contrast, both NF-kappaB and VEGF-C were more commonly detected in infiltrating ductal carcinoma than in intraductal carcinoma and normal tissue. Fas expression was correlated with tumor size, histological grade, and clinicopathological stage; NF-kappaB expression was correlated with tumor size, histological grade, lymph node metastasis; VEGF-C expression was correlated with lymph node metastasis and clinical and pathological stages of breast cancer (p < or = 0.05). Spearman rank correlation analysis revealed a negative correlation between Fas expression and both NF-kappaB and VEGF-C expression in infiltrating breast cancer (p < 0.05) Additionally, Kaplan-Meier survival analysis demonstrated that five-year survival was higher for patients with Fas-positive samples but lower for those with VEGF-C-positive samples. CONCLUSIONS: The present results demonstrate that Fas and NF-B play a role in the initiation and development of breast cancer, while VEGF-C appears to promote lymph node metastasis. Thus, these proteins may serve as useful diagnostic and prognostic markers of invasive breast cancer.

Enhanced radioimmunotherapeutic efficacy of a monoclonal antibody cocktail against SMMC-7721 human hepatocellular carcinoma.

The improved tumoricidal effect of the radioantibody mixture ("cocktail") has been reported recently for the treatment of colon tumor. In the present study, we demonstrated the enhanced radioimmunotherapeutic efficacy of a monoclonal antibody (MAb) cocktail against human hepatocellular carcinoma. Therapeutic efficacy was determined by measuring the change in tumor size over a period, determining the percentage of growth inhibition of each treatment at various times after radioantibody therapy. Radioimmunotherapy of SMMC-7721 human hepatoma xenografts in athymic nude mice with combination of 131I-labeled Hepama-1 and 131I-labeled 9403 mouse MAbs was more effective than using either Hepeam-1 or 9403 MAb alone The MAb cocktail could target a greater number of hepatoma cells and increase the magnitude of hepatoma cell uptake of radioantibodies. The in vitro results explain the enhanced effect of the MAb cocktail in in vivo model system.

[Gas chromatographic analysis of aldicarb and its metabolites in urine].

A method is described for the analysis of aldicarb and its metabolites in urine by GC/FPD. The sample was concentrated with activated charcoal-Florisil column chromatography, eluted by dichloromethane-acetone (1:1v/v). The aldicarb, aldicarb sulfoxide in the eluted solution were oxidized by oxidizing reagent into aldicarb sulfone. The concentration of aldicarb sulfone was analyzed by GC/FPD. The detection limit of 0.0024 mg/L and coefficient of variation of 2.4%-7.4% were achieved. Mean recovery rates were 90.9%, 86.6%, 92.6% for aldicarb, aldicarb sulfoxide and aldicarb sulfone, respectively.

Immunohistochemical localization of neuropeptide Y in the guinea pig medulla oblongata. Correlation with VIP and DBH.

The immunohistochemical localization of neuropeptide Y (NPY) was correlated with those of dopamine-beta-hydroxylase (DBH) and vasoactive intestinal polypeptide (VIP) by mapping serial 7 micron paraffin sections at three levels of the guinea pig lower brainstem: a) area postrema, b) dorsal motor nucleus of the vagus, and c) nucleus prepositus of the hypoglossal nerve. Based on differences in transmitter expression, three populations of NPY-immunoreactive (IR) neurons were distinguished: NPY-IR catecholaminergic cells (NPY/CA), NPY-IR VIP-ergic cells (NPY/VIP), and NYP-IR cells which were not reactive to either DBH or VIP. Within these populations, size differences among neurons in characteristic locations allowed differentiation among the following subpopulations: NPY/CA neurons in the lateral reticular nucleus--magnocellular part (mean neuronal size 538 micron2) and parvocellular part (318 micron2)-, in the vagus-solitarius complex (433 micron2), and in the dorsal strip (348 micron2); NPY/VIP neurons in the vagus-solitarius complex (368 micron2) and in the nucleus ovalis (236 micron2). Apart from scattered NPY-IR cell bodies in the regions listed above, NPY-IR cell bodies in the lateral portion of the nucleus solitarius and in the caudal part of the spinal nucleus of the trigeminal nerve did not exhibit IR to either DBH or VIP. NPY-IR neurons in the area postrema occurred too infrequently for co-localization studies. The differential distribution of heterogeneous NPY-IR cell subpopulations may reflect the involvement of NPY in a variety of neuronal functions.