Hemostatic Effect of Hemocoagulase Agkistrodon on Surgical Wound in Breast Cancer Surgery.

Objective To evaluate the hemostatic effect of hemocoagulase agkistrodon on surgical wound in breast cancer surgery. Methods Totally 60 patients undergoing breast cancer surgery were enrolled in this prospective,randomized,double-blinded,and controlled study. All the patients met the inclusion and exclusion criteria and signed the informed consent. Hemocoagulase agkistrodon (2 U) was injected 20 minutes before surgery and 4 and 24 hours after surgery in the intervention group (n=30),whereas normal saline was used instead in the control group (n=30). The volume of intraoperative bleeding,wound drainage volume 1-3 days after surgery,and total drainage volume were recorded. Meanwhile,the change of blood coagulation function,treatment safety,and clinical outcomes were observed. Results The intra-operative hemorrhage volume of the intervention group [(95.0±48.3)g] was significantly lower than that of the control group [(144.8±105.4)g] (t=-2.07,P=0.044). The volume of total drainage of the intervention group [(166.7±71.2)g] was significantly lower than that of the control group [(251.4±166.3)g] (t=-2.29,P=0.029). The hemoagglutination indicators were similar in the two groups and no complication such as thrombosis occurred. The length of hospital stay of the intervention group [(15.00±3.53)d] was similar to that of the control group [(15.92±2.32)d] (t=-1.057,P=0.297). No research drug-related adverse event was occurred in our study. Conclusion Hemocoagulase agkistrodon has good hemostatic effect for patients undergoing breast cancer surgery without increasing the risk of thrombosis.

[The New Bacteria Expressing Recombinant Multi-epitope Vaccine against Helicobacter pylori and Its Microbiological Characteristics].

OJECTIVE: To construct the engineering bacteria with recombinant plasmid expressing the multi-epitope vaccine which composed of Helicobacter pylori urea membrane channel protein (UreI), Helicobacter pylori urease B subunit (UreB) and cholera toxin B subunit (CTB), and then to study it's microbiological characteristics. METHODS: The sequence contains some dominant epitopes of Helicobacter pylori UreI and UreB was designed, and ctB was added at the N-terminal, all the sequence were linked by flexible linkers. Codon optimization was done according to Escherichia coli (E. coli) BL21 (DE3) bias, the optimized sequence was designated BIB. BIB sequence was synthesized and cloned into plasmid pET28a(+). The recombinant plasmid was confirmed by restriction enzyme digestion and DNA sequencing. The recombinant protein BIB was expressed in E. coli BL21 (DE3) and analyzed by Western blot. RESULTS: The plasmid of pET28a(+)/BIB was constructed successfully, confirmed by restriction enzyme digestion and DNA sequencing. The recombinant protein BIB with relative molecular mass about 33 x 10(3) could be produced by E. coli BL21 (DE3) and was detected by Western blot. The relative molecular mass and N-terminal amino acid sequence of BIB were 100% identity with the design. CONCLUTION: The engineering bacteria with recombinant plasmid expressing the multi-epitope vaccine against Helicobacter pylori was constructed successfully. The recombinant protein BIB can be identified by anti-Sydney strain 1 of Helicobacter pylori (H. pylori SS1) polyclonal antibody and anti-CTB monoclonal antibody, which demonstrated that BIB has the expected antigenicity.

[Association of bacterial diversity in city area of Nanming river with environmental factors].

OBJECTIVE: The aim of this study was to analyze the bacterial diversity of urban section of Nanming river as well as to study relationship between the bacterial diversity and environmental factors. METHODS: The high throughput sequencing on PCR-amplified 16S rDNA V4 fragments was used to determine the bacterial diversity of samples from five sites of Nanming river. The ordination technique of redundancy analysis was used to analyze the effects of the environmental factors on bacterial community composition. RESULTS: Diversity index analysis of bacterial community composition in Nanming river showed that the Shannon-Wiener index of bacterial diversity in Nanmng River was about 7.5. The Shannon-Wiener index of 5 sampling site had an order as Wudang bridge > Shuikou Si > Wuyan bridge > Huaxi bridge > Guanzhou bridge. Based on the sequencing results, there were 11 phyla (327 genera) in the samples, of which proteobacterice (66.1% ± 3.30% ) was the dominant phyla, Gamma proteobacteria (54.76% ± 4.86%) was the dominant subgroup and Pseudomonas (16.92% ± 0.02%) was the dominant genera. In addition, there are some flora and rare flora undetermined. The result of RDA suggested that the influence of different environmental factors on different microbes were different. Bacterial community IV had significant positive correlation with total nitrogen and total phosphorus in the environment. CONCLUSION: This research provides references for the association of bacterial composition and diversity with environmental factors.

Facile colorimetric sensor using oxidase-like activity of octahedral Ag2O particles for highly selective detection of Pb(II) in water.

Pb(II) is a prevalent heavy metal ion classified as a 2B carcinogen. Excessive intake of Pb(II) in the human body can damage the central nervous system, kidneys, liver, and immune system, leading to permanent brain damage, anemia, and cancer. Colorimetry can be applied to rapidly determine Pb(II) residues, but there are still many challenges in the accuracy and sensitivity of detection. Based on the inhibitory impact of Pb(II) on the oxidase-like activity of octahedral silver oxide (Ag2O), a colorimetric sensor with smartphone-assisted analysis for the Pb(II) detection was first developed. Herein, it has been found that Pb(II) can adsorb onto the surface of octahedral Ag2O, hindering the production of O2- in the reaction system. This ultimately results in the suppression of oxidase-like activity, leading to a lighter purple appearance of the colorimetric reaction solution. The sensor exhibits a high degree of sensitivity and a limit of detection (LOD) for Pb(II) was calculated as 2.2 µg L-1. Hence, the developed colorimetric sensor with high sensitivity, excellent specificity, and high tolerance to sodium ions is hopeful to have practical applications in Pb(II) detection in environmental water samples. Moreover, the sensor will provide a novel strategy for heavy metal ion detection and other substances.

A stable colorimetric biosensor for highly selective detection of malathion residue in food based on aptamer-regulated laccase-mimic activity.

Malathion causes a serious threat to human health due to its widespread use in the environment. Herein, a novel and stable smartphone-integrated colorimetric biosensor for malathion detection is firstly established based on aptamer-enhanced laccase-mimicking activity. The results indicate that the M17-F aptamer can increase the affinity of Ag2O nanoparticles to the substrate 2,4-dichlorophenol and enhance their laccase-mimicking activity. Thus, abundant semiquinone radicals are produced in the catalytic system, which are combined with chromogenic agent to generate dark red products. The corresponding RGB values for the colour change of the solution can be easily obtained using smartphones, which is used for the rapid detection of malathion. The established biosensor for malathion has a limit of detection as low as 5.85 nmol·L-1, and displays good selectivity for other competitive pesticides. Moreover, further studies have verified the applicability of the biosensor in actual samples, indicating that it may have the potential for application in malathion detection in food.

Rapid colorimetric sensor for ultrasensitive and highly selective detection of Fumonisin B1 in cereal based on laccase-mimicking activity of silver phosphate nanoparticles.

Ag3PO4 nanoparticles (NPs) was prepared through a facile coprecipitation method, and was first found to have excellent laccase-mimicking catalytic activity. The study confirms that Fumonisin B1 (FB1) can effectively hinder the production of superoxide anion (O2-) between Ag3PO4 NPs and dissolved oxygen, and further inhibit laccase-mimicking activity of Ag3PO4 NPs. Thus, a novel rapid colorimetric sensor for FB1 analysis in cereal was first established using laccase-mimicking activity as sensing signal. The absorbance variation of sensing solution is directly related to the amount of FB1, and the color change is further combined with smartphone for quantitively analysis of FB1. The limit of detection (LOD) of the sensor is determined as low as 1.73 µg·L-1, which is far lower than the maximum residue limits (MRLs) of FB1 set by European Commission and US Food and Drug Administration (FDA). The average recovery of 87.8-104.5% for FB1 detection was obtained in cereal.

Porous-nanozyme-based colorimetric sensor for rapid detection of kanamycin in foods under neutral condition.

Most colorimetric methods based on nanozymes need to have excellent performance under acidic condition, so they are currently facing some challenges in the field of food hazard detection. Herein, a facile and rapid colorimetric sensor for kanamycin (KAN) detection in foods under neutral condition has been designed using the peroxidase-mimic activity of porous nanozyme like Co3 O4 nanodisk. Further investigations showed that the interaction mechanism between porous Co3 O4 nanodisk and substrates belongs to a ping-pong model, and the inhibition type of KAN on the peroxidase-mimic activity is noncompetitive inhibition. The constructed sensor has good sensitivity for KAN with the limit of 57 nM, and the color changes can be discerned visually when KAN exceeds 0.5 µM. Besides, the colorimetric sensor obtains excellent recovery results in chicken serum, milk, honey, and pork, which shows that the proposed sensing strategy can provide a rapid and convenient detection method for KAN in foods under neutral condition. PRACTICAL APPLICATION: The established sensing strategy can rapidly distinguish whether KAN residue exceeds the permissible level within 10 min, which meets the requirement for on-site monitoring of antibiotics in foods, and also open up a new idea for other hazards detection under neutral condition.

Aptamer-enhanced the Ag(I) ion-3,3',5,5'-tetramethylbenzdine catalytic system as a novel colorimetric biosensor for ultrasensitive and selective detection of paraquat.

A facile and simple colorimetric biosensor was first established for paraquat (PQ) detection based on the aptamer-enhanced oxidation process of 3,3',5,5'-tetramethylbenzidine (TMB) by Ag+. The study confirmed that the interaction of PQ-15 aptamer with Ag+ accelerates the electron transfer from the aptamer-Ag+ complex to dissolved oxygen, which enhances the release of superoxide anion radicals (O2̇-) and facilitates the catalytic oxidation of the chromogenic substrate. PQ-15 aptamer will preferentially bind to PQ molecules, resulting in no further enhancement of the catalytic activity of Ag+. Molecular docking results revealed that the PQ molecules are attached to the stem-loop region of the PQ-15 aptamer through σ-π conjugation interactions. The proposed method is simple that only contains Ag+ and corresponding aptamer. The limit of detection (LOD) of the constructed colorimetric biosensor for PQ detection was determined to be 16.5 µg·L-1, belowing the maximum residue limit in fruits and vegetables set by the EU. Moreover, the colorimetric biosensor showed excellent selectivity and anti-interference properties, which was validated for detecting PQ residues in several typical agricultural and water samples.

Gut bacteria reflect the adaptation of Diestrammena japanica (Orthoptera: Rhaphidophoridae) to the cave.

The gut microbiota is essential for the nutrition, growth, and adaptation of the host. Diestrammena japanica, a scavenger that provides energy to the cave ecosystem, is a keystone species in the karst cave in China. It inhabits every region of the cave, regardless of the amount of light. However, its morphology is dependent on the intensity of light. Whether the gut bacteria reflect its adaptation to the cave environment remains unknown. In this research, D. japanica was collected from the light region, weak light region, and dark region of three karst caves. The gut bacterial features of these individuals, including composition, diversity, potential metabolism function, and the co-occurrence network of their gut microbiota, were investigated based on 16S rRNA gene deep sequencing assay. The residues of amino acids in the ingluvies were also evaluated. In addition, we explored the contribution of gut bacteria to the cave adaptation of D. japanica from three various light zones. Findings showed that gut bacteria were made up of 245 operational taxonomic units (OTUs) from nine phyla, with Firmicutes being the most common phylum. Although the composition and diversity of the gut bacterial community of D. japanica were not significantly different among the three light regions, bacterial groups may serve different functions for D. japanica in differing light strengths. D. japanica has a lower rate of metabolism in cave habitats than in light regions. We infer that the majority of gut bacteria are likely engaged in nutrition and supplied D. japanica with essential amino acids. In addition, gut bacteria may play a role in adapting D. japanica's body size. Unveiling the features of the gut bacterial community of D. japanica would shed light on exploring the roles of gut bacteria in adapting hosts to karst cave environments.

The complete mitochondrial genome of Melanostoma orientale (Diptera: Syrphidae).

The mitochondrial genome of Melanostoma orientale has been decoded by Illumina sequencing. The mitogenomic size is 16,229 bp with 40.96% A, 40.29% T, 10.60% C, and 8.15% G. It is encoded with 13 protein-coding genes, 2 ribosomal RNA, and 22 transfer RNAs. The phylogenetic tree showed that 10 species of Syrphidae, belonging to six genera, were clustered into two clades. This is the first mitochondrial genome for the genus Melanostoma.

The complete mitochondrial genome of Eysarcoris guttigerus (Hemiptera: Pentatomidae).

The mitochondrial genome of Eysarcoris guttigerus has been decoded using Illumina sequencing. The mitogenomic size is 15,368 bp with 42.97% A, 33.90% T, 13.22% C, and 9.90% G. It encoded 13 protein-coding genes, two ribosomal RNA, and 22 transfer RNAs. Phylogenetic tree showed that genus Eysarcoris had a closer relationship with the genus Carbula than the rest of 20 genera.

MicroRNA-361-3p promotes human breast cancer cell viability by inhibiting the E2F1/P73 signalling pathway.

Analysis of the microRNA (miRNA) expression signature of breast cancer based on RNA sequencing demonstrated that miR-361-3p was significantly upregulated in breast cancer tissues. miR-361-3p is a novel miRNA, and its role in breast cancer is currently unclear. The aim of the present study was to investigate the functions of miR-361-3p in breast carcinoma. In this study, it was observed that the expression of miR-361-3p in cancer tissues was significantly higher compared with that in para-cancerous tissues and was correlated with advanced TNM stage, Ki-67 overexpression and shorter disease-free survival. Overexpression of miR-361-3p promoted proliferation and inhibited apoptosis of breast cancer cells. Through RNA sequencing, multi-library retrieval, luciferase reporter assays, quantitative polymerase chain reaction analysis, western blotting and other methods, it was verified that E2F1 was directly downregulated by miR-361-3p. The knockdown of E2F1 by siRNA promoted breast cancer cell proliferation and inhibited apoptosis, similar to miR-361-3p. In addition, miR-361-3p was able to decrease the expression of P73 by targeting E2F1, whereas overexpression of P73 reversed the effect of miR-361-3p on the viability of breast cancer cell lines. Thus, the present study demonstrated that miR-361-3p acts as an oncomiR in breast cancer to promote proliferation and inhibit apoptosis through inhibiting the P73 pathway by downregulating E2F1 expression, which may uncover valuable prognostic factors or treatment targets.

Epidural Analgesia Improves Postoperative Pain Control but Impedes Early Discharge in Patients Undergoing Pancreatic Surgery.

OBJECTIVES: The aim of this study was to evaluate the impact of epidural analgesia (EA) on postoperative length of stay (LOS), expeditious discharge, and pain relief after pancreaticoduodenectomy (PD) and distal pancreatectomy (DP). METHODS: Retrospective reviews of 2014-2015 American College of Surgeons National Surgical Quality Improvement Program databases and our institutional pancreatic surgery database were conducted. RESULTS: On univariate analysis, EA was associated with statistically significant longer lengths of stay for both PD and DP. On comparative analysis at mode LOS, discharged before versus after 7 days for PD and 6 days for DP, EA was a significant predictor for the longer groups for both procedures on multivariable analysis (PD, odds ratio of 1.465, P < 0.001; DP, odds ratio of 1.471, P = 0.004). On review of our institution's pancreatic surgery database, patient-reported pain scores were significantly lower in the EA groups than intravenous narcotics groups on the day of surgery only for both PD and DP. CONCLUSIONS: Epidural analgesia was associated with longer LOS with a most pronounced effect on early discharge after surgery for patients undergoing open PD and DP. It only resulted in superior pain control on the day of surgery.

Protection Against Helicobacter pylori Infection in BALB/c Mouse Model by Oral Administration of Multivalent Epitope-Based Vaccine of Cholera Toxin B Subunit-HUUC.

Vaccination is an increasingly important alternative approach to control Helicobacter pylori infection, since H. pylori resistance to previously efficacious antibiotic regimens is increased, and H. pylori eradication treatment for upper gastrointestinal diseases is becoming less successful. Fortunately, an efficient oral monovalent H. pylori vaccine has been developed. However, compared with monovalent vaccines, multivalent vaccines have the potential to induce more effective and comprehensive protection against H. pylori infection. In this study, we designed and produced a multivalent epitope-based vaccine cholera toxin B subunit (CTB)-HUUC with the intramucosal adjuvant CTB and tandem copies of B-cell epitopes (HpaA132-141, UreA183-203, and UreB321-339) and T-cell epitopes (HpaA88-100, UreA27-53, UreB229-251, UreB317-329, UreB373-385, UreB438-452, UreB546-561, CagA149-164, and CagA196-217) from H. pylori adhesion A subunit (HpaA), urease A subunit (UreA), urease B subunit (UreB), and cytotoxin-associated antigen (CagA). Serum IgG, stomach, and intestine mucosal sIgA from mice after CTB-HUUC vaccination neutralized H. pylori urease activity in vitro. CTB-HUUC vaccination promoted H. pylori-specific lymphocyte responses and a mixed CD4+ T cell immune response as indicated by IFN-γ, interleukin-4, and interleukin-17 production in mice. Both oral prophylactic and therapeutic CTB-HUUC vaccinations reduced gastric urease activity and H. pylori infection and protected stomachs in mice. Taken together, CTB-HUUC is a promising potent and safe multivalent vaccine in controlling H. pylori infection in BALB/c mouse model.

Micro-Incision of the Cystic Duct Confluence in Laparoscopic Common Bile Duct Exploration for Elderly Patients with Choledocholithiasis.

Common bile duct (CBD) stones are common in elderly patients. The laparoscopic transcystic approach with micro-incision of the cystic duct confluence in common bile duct exploration (LTM-CBDE) is a modified laparoscopic transcystic approach. Its safety and efficacy have not been studied in elderly patients with secondary choledocholithiasis. This study evaluates the safety and efficacy of LTM-CBDE in elderly (≥65 years) patients with secondary choledocholithiasis and compares the results with those in younger patients. In this retrospective analysis, 128 patients underwent LTM-CBDE from March 2007 to December 2013. The patients were divided into two groups according to age: the elderly group consisted of 50 patients aged ≥65 years and the younger group consisted of 78 patients aged <65 years. The preoperative morbidity rate, American Society of Anesthesiologists (ASA) score, previous abdominal operations, operation time, postoperative hospital stay, open conversion rate, postoperative complication rate, residual stone rate, recurrence rate and mortality were compared in both groups. The preoperative morbidity (41 vs. 28) and ASA score (2.5 ± 0.7 vs. 1.8 ± 0.6) were higher in the elderly group (P = 0.000, in both groups). No significant differences in previous abdominal operations, operation time, postoperative hospital stay, open conversion rate, postoperative complication rate, residual stone rate, recurrence rate and mortality (P > 0.05) were found between the two groups from March 2007 to December 2013. LTM-CBDE is a safe and effective treatment procedure for elderly patients with secondary choledocholithiasis. For suitable patients, we recommend LTM-CBDE as the treatment of choice.

AFP mRNA level in enriched circulating tumor cells from hepatocellular carcinoma patient blood samples is a pivotal predictive marker for metastasis.

Circulating tumor cells (CTCs) quantification may be helpful for evaluating cancer dissemination, predicting prognosis and assessing therapeutic effectiveness and safety. In the present study, CTCs from blood samples of 72 patients with hepatocellular carcinoma (HCC) were enriched with anti-EpCAM nanoparticles. AFP mRNA level was detected by nested polymerase chain reaction (PCR) after enrichment of CTCs from HCC blood samples at 0, 3, 6, 9 and 12 months after hepatectomy, respectively. AFP mRNA expression in CTCs was positive in 43 patients (59.7%) and negative in 29 patients (40.3%) before hepatectomy. Among 43 patients with positive AFP mRNA expression in CTCs before hepatectomy, 10 and 11 were diagnosed as intrahepatic/extrahepatic metastasis before and after hepatectomy, respectively. In addition, these 21 patients with metastasis had persisting positive AFP mRNA of CTCs during the whole tested year. Specifically, 3 patients with AFP mRNA negative in CTCs before hepatectomy changed to be positive at 6 and 9 months, and 2 of them were diagnosed as metastasis 12 months after hepatectomy. We conclude that the positive AFP mRNA of CTCs can be a pivotal predictor for HCC metastasis before and after hepatectomy. The release of AFP expression from hepatocellular carcinoma cells into circulation must be a major source of HCC metastasis.

Upconversion fluorescence-SERS dual-mode tags for cellular and in vivo imaging.

Fluorescent-surface enhanced Raman scattering (F-SERS) dual mode tags showed great potential for bioimaging due to the combined advantages of intuitive, fast imaging of fluorescence and multiplex capability of SERS technique. In previously reported F-SERS tags, organic fluorescent dyes or quantum dots were generally selected to generate fluorescence signal. Herein, we reported the first proof-of-concept upconversion fluorescence (UCF)-SERS dual mode tags based on near infrared (NIR) laser (980 nm) excited upconversion nanoparticles (UCNPs) for live-cell and in vivo imaging. Three components involved in this tag: NaYF4:Yb,Er UCNPs@SiO2 serving as the fluorescent core of the tag; silver nanoparticles in situ grown on the surface of UCNPs@SiO2 for generating characteristic Raman signal; and denatured BSA coating rendering the tag's stability and biocompatibility. The UCF-SERS tags integrated the NIR imaging capability of both fluorescent UCNPs and plasmonic SERS nanoprobe, which facilitated dual mode bioimaging investigation, especially for living animals. Ex vivo experiments revealed that with 980 nm and 785 nm NIR laser irradiations, the UCF and SERS signals of the tags could be detected from 3 and 7 mm deep pork tissues, respectively. Furthermore, the in vivo imaging capabilities of UCF-SERS tags were successfully demonstrated on living mice. The developed dual modality tags held great potential for medical diagnostics and therapy.