[Comparison of early postoperative effects between anterior bladder and posterior bladder approaches in robotic assisted laparoscopic radical prostatectomy].

Objective: To examine the effect of robotic assisted laparoscopic radical prostatectomy with different approaches on early postoperative effects. Methods: Totally 44 patients (average age of 65.9 years, range: 46 to 81 years) underwent robotic assisted laparoscopic radical prostatectomy by a single operator at Department of Urology, Peking Union Medical Collage Hospital from March 2018 to March 2020 were retrospectively analyzed. The mean age was 65.9 years (range: 46 to 81 years), including 24 cases in the anterior bladder approach group (anterior approach group) and 20 cases in the posterior bladder approach group (posterior approach group). The preoperative clinical data, perioperative related data and postoperative urinary control recovery were compared between the two groups by t test, χ2 test or Fisher exact test. Results: In terms of clinical data, there was no difference in age, prostate volume, preoperative prostate specific antigen and Gleason score(all P>0.05). There was no significant difference in operation time ((184±43) minutes vs. (193±42) minutes, t=-0.599, P=0.55), bleeding volume ((218±88) ml vs. (225±115) ml, t=-0.244, P=0.81), postoperative stage (T2/T3: 15/9 vs. 12/8, χ²=0.029, P=0.87) and positive rate of cutting edge (29.2% (7/24) vs. 30.0% (6/20), χ²=0.004, P=0.95). In terms of postoperative urinary control, patient rates who achieved urinary control immediately after extubation was significantly higher for the posterior approach group than the anterior approach group (30.0% (6/20) vs. 4.2% (1/24), P=0.04). There was no significant difference between two groups for those who achieved urinary control 3 months after operation (6 cases vs. 11 cases, P=0.06), 6 months after operation (20 cases vs. 19 cases, P=0.36) and those who achieved urinary control 12 months after operation (23 cases vs. 19 cases, P=1). Conclusion: For robotic assisted laparoscopic radical prostatectomy, the posterior approach does not prolong the operation time, does not increase the amount of bleeding, and improves the short-term postoperative urinary control.

[Surgery treatment of ectopic adrenocorticotrophic hormone syndrome].

OBJECTIVE: To investigation the diagnosis and treatment of ectopic adrenocorticotrophic hormone (ACTH) syndrome. METHODS: The clinical characters of 57 cases of ecotopic ACTH syndrome from Jan. 1996 to Dec. 2016 were collected and analyzed. The 57 cases included 32 males and 25 females. The age ranged from 11 to 68 years (average 32 years). ACTH levels significantly increased from 16.5 to 365.6 pmol/L, with average 77.6 pmol/L (normal range <10.1 pmol/L). The pituitary MRI did not found lesions. The CT showed that their bilateral adrenal glands diffused small nodular changes or nodular hyperplasia. The 57 cases were divided into 3 groups according to different treatment options. In the study, 25 ectopic ACTH syndrome cases (44%) were group A, without identified source of ectopic hormone, were treated with bilateral or unilateral adrenalectomy due to the severity of the disease and difficulty of operation. Group B was composed of 16 cases (28%) diagnosed as ectopic ACTH syndrome by finding ectopic ACTH tumors and surgical resection. Group C included 16 cases (28%) with nonsurgical therapy. Different treatment results and prognosis were analyzed. RESULTS: In the study, 40 cases of the 57 had been followed up for 6 months to 10 years. In group A, of the 25 cases with bilateral or unilateral adrenalectomy, 4 died of diabetes and severe pulmonary infection, 18 survived, and 3 were lost to the follow-up, and the survival rate was 81% (18/22). In group B, of the 16 cases with radical tumor resection, 5 died of tumor recurrence 0.5-6.0 years after operation, 3 survived, and 8 were lost to the follow-up, and the survival rate was 37.5% (3/8). In group C, of the 16 non-operation patients, 4 with radiotherapy and chemotherapy died of metastases, diabetes or pulmonary infection, 6 with chemotherapy died of pulmonary infection within 1 year and the others were lost to the follow-up, and the survival rate was 0. CONCLUSION: Ectopic ACTH syndrome is difficult to treat. Adrenalectomy is effective for the management of ectopic ACTH syndrome, especially for those patients with severe Cushing's syndrome, but the primary tumor can not be located.

Characteristics of cathelicidin-Bg, a novel gene expressed in the ear-side gland of Bufo gargarizans.

The traditional Chinese medicine Chan Su (toad venom) comprises dried secretions of the ear-side gland of Bufo gargarizans. Chan Su is known for its small molecular components, which include telocinobufagin, marinobufagin, and bufalin, while in other amphibians, studies mainly focus on peptide components. Until recently, no genes expressed in the ear-side gland of B. gargarizans gland had been cloned. In this study, cathelicidin-Bg, a coding sequence of anti-microbial peptide (AMP), was cloned. The predicted amino acid sequence of cathelicidin-Bg was very similar to that from other amphibians, with a 34-amino acid mature peptide predicted in the C-terminus. The functions of this mature peptide were verified by microbe and tumor cell inhibition assays. Our results showed that the mature peptide of cathelicidin-Bg could inhibit the proliferation of Staphylococcus aureus and Pseudomonas aeruginosa. The mature peptide was also shown to selectively inhibit tumor cells. These results indicate that the identified coding sequence represents an active peptide of Chan Su.

The main functions and structural modifications of tripeptide N-formyl-methionyl-leucyl-phenylalanine (fMLP) as a chemotactic factor.

Gram negative bacteria-derived and synthetic N-formyl peptides play a key role in host defense as chemotactic factors for phagocytic leukocytes. The first compound to be identified was N-formylmethionyl-leucyl-phenylalanine (fMLP) which contains highly potent leukocyte chemoattractant. Natural fMLP was subsequently purified and identified in supernatants of gram negative bacteria. Recently, much more attention has been focused on the human formyl peptide receptor (FPR) and its variant formyl peptide receptor-like 1 (FPRL1) and formyl peptide receptor-like 2 (FPRL2). Chemotactic factors such as fMLP interact with their specific cell surface receptors, which results in multiple biological responses through a G protein-coupled signal pathway. In this review, the functions and structural modifications of fMLP are discussed in view of future drug development.

Recent advances in P-glycoprotein-mediated multidrug resistance reversal mechanisms.

Expression of the multidrug resistance (MDR) phenotype is responsible for chemotherapy failure in numerous cancers. Overexpression of mdr1 gene-encoded permeability glycoprotein (P-gp) is known to play a pivotal role in the development of this phenotype. The role of P-gp has been proposed as an important goal in the design of chemotherapy strategies. However, modulation of P-gp activity by chemotherapy has limited possibilities because of toxicity and poor specificity. In this article, we review the latest advancements in different potential P-gp-mediated MDR reversal mechanisms as well as the methods of evaluating MDR reversal activity, which would be helpful in finding novel MDR reversal agents (or chemosensitizers).

Metabolic maturation of the brain: a study of local cerebral glucose utilization in the developing cat.

Previously, using positron emission tomography (PET), we showed that local cerebral metabolic rates for glucose (lCMRglc) in children undergo dynamic maturational trends before reaching adult values. In order to develop an animal model that can be used to explore the biological significance of the different segments of the lCMRglc maturational curve, we measured lCMRglc in kittens at various stages of postnatal development and in adult cats using quantitative [14C]2-deoxyglucose autoradiography. In the kitten, very low lCMRglc levels (0.14 to 0.53 mumol min-1 g-1) were seen during the first 15 days of life, with phylogenetically older brain regions being generally more metabolically mature than newer structures. After 15 days of age, many brain regions (particularly telencephalic structures) underwent sharp increases of lCMRglc to reach, or exceed, adult rates by 60 days. This developmental period (15 to 60 days) corresponds to the time of rapid synaptic proliferation known to occur in the cat. At 90 and 120 days, a slight decline in lCMRglc was observed, but this was followed by a second, larger peak occurring at about 180 days, when sexual maturation occurs in the cat. Only after 180 days did lCMRglc decrease to reach final adult values (0.21 to 2.04 mumol min-1 g-1). In general, there was good correlation between the metabolic maturation of various neuroanatomical regions and the emergence of behaviors mediated by the specific region. At least in the kitten visual cortex, which has been extensively studied with respect to developmental plasticity, the "critical period" corresponded to that portion of the lCMRglc maturational curve surrounding the 60-day metabolic peak. These normal maturational lCMRglc data will serve as baseline values with which to compare anatomical and metabolic plasticity changes induced by age-related lesions in the cat.

PSMA mimotope isolated from phage displayed peptide library can induce PSMA specific immune response.

Prostate-specific membrane antigen (PSMA) is a cell surface glycoprotein expressed predominantly in prostate secretory acinar epithelium and prostate cancer cells as well as in several extraprostatic tissues. Mouse monoclonal antibody 4G5 specific to the extracellular domain of PSMA was used to screen two phage displayed peptide libraries (9aa linear and 9aa cys library). Three 4G5-reactive phagotopes were identified. Sequence analysis of isolated clones demonstrated that the interaction motif "VDPA/SK" has high homology to 719-725aa on PSMA. Immunohistochemical staining of the prostate cancer sample with the PSMA-mimic phagotope (mimotope) immunized serum antibodies demonstrate that the mimotope isolated from the phage displayed peptide libraries can induce PSMA specific immune response in vivo.

cDNA and genomic structures of arrowhead proteinase inhibitors.

Arrowhead double-headed proteinase inhibitors A and B consist of 150 amino acid residues with three disulfide bonds. Based on their primary structures, three cDNA fragments of the inhibitors were amplified in vitro by the PCR method using a constructed arrowhead cDNA library as a template. With the overlapping sequences, the full-length cDNA sequences of the inhibitors were then ascertained. The open reading frame encodes a pre-inhibitor, including a 24 residue signal peptide. The deduced amino acid sequences are consistent in principle with those determined by primary structure analysis, except that there are seven extra residues in the C-terminal part of the inhibitors, which might be cleaved off by proteinase post-processing immediately after protein synthesis. It is worth pointing out that cDNAs of both inhibitors A and B contain an 87 bp intron in the AAG codon of residue Lys-97. According to the elucidated cDNA sequences, the structural genes of inhibitors A and B were amplified using the total cDNA or genomic DNA of arrowhead as a PCR template. It was indicated that both the cDNA and genomic structures of inhibitors A and B have the same sequences.

Electron microscopy study on a new phase in beta-titanium alloys aged at a high temperature.

In a Ti-20mass%Mo alloys aged by two steps aging at 623 and 823K, a new phase has been observed. In the first step aging, [Formula: see text] -phase crystals appear and at the second step aging, these crystals disappear and are replaced with the newly discovered phase crystals. Observations in the dark field image and analysis of composition of the new phase crystals have shown that the formation is closely related to the [Formula: see text] -phase structure. The new phase crystals have been also observed in four kinds of [Formula: see text] -type Ti alloys and a ( [Formula: see text] ) type Ti alloy aged at high elevated temperatures. Based on high-resolution electron microscope observations, the atomic structure of the new phase is assumed.

HDAC6: physiological function and its selective inhibitors for cancer treatment.

Acetylation and deacetylation of histones are important in regulating gene expression and play a key role in modification of gene transcription. Specific HDACs isoforms can be regarded as a target for cancer therapy avoiding side-effects, HDAC6 with a unique physiological function and structure has become a hot issue recently. The unique isoform HDAC6 is involved in tumorigenesis, development and metastasis through tubulin, HSP90, invasin and ubiquitin-protein. Here we review the structure elements, biological function, and recent selective inhibitors of HDAC6, and study the structure-activity and structure-selectivity relationship.

A new boronic acid-based fluorescent sensor for L-dihydroxyphenylalanine.

Catecholamines, such as dopamine and L-dihydroxyphenylalanine (L-DOPA), are associated with different physiological functions and diseases. In our recent studies, a novel water-soluble boronic acid compound 3c was identified as a selective fluorescent sensor for L-DOPA. This compound not only has the ability to interact with dopamine and catechol, but also has no fluorescence intensity change for L-DOPA precursors in vivo, such as L-tyrosine.

Design, synthesis and activity study of aminopeptidase N targeted 3-amino-2-hydroxy-4-phenyl-butanoic acid derivatives.

A series of (2RS,3S)-3-amino-2-hydroxy-4-phenyl-butanoic acids (AHPA) derivatives (MA0-MA7) were synthesized. The in vitro aminopeptidase N (APN) enzyme and cell proliferation assay of target compounds were investigated. The results showed that most compounds displayed potent inhibitory activities against APN, compound MA0 showed even better inhibitory effects than bestatin on both enzyme activity and HL60 cell proliferation. The FlexX docking result showed the mode of binding between MA0 and APN.

Expression, purification, and S-nitrosylation of recombinant histone deacetylase 8 in Escherichia coli.

Histone deacetylase (HDAC) 8 is a zinc ion dependent enzyme involved in removing the acetyl group from the core histones and other proteins which belong to Class I HDACs. It was reported that nitric oxide (NO) is a key regulator of HDAC function and S-nitrosylation of HDAC2 induces chromatin remodelling in neurons. This work reports the successful recombinant expression of human HDAC8 in Escherichia coli with two plasmids and the purification and S-nitrosylation in vitro. It was found that HDAC8 can be S-nitrosylated by the NO donor S-nitrosoglutathione (GSNO) in vitro, and the activity of HDAC8 was significantly inhibited when incubated with GSNO and S-nitrosocysteine in a time- and dosage-dependent manner, but sodium nitroprusside (SNP), and dithiothreitol cannot reverse this inhibition. These observations support and extend the concept that NO may regulate HDAC8 function by S-nitrosylation.

Direct effects of fascaplysin on human umbilical vein endothelial cells attributing the anti-angiogenesis activity.

Novel anti-angiogenesis activity of fascaplysin via VEGF blockage was recently revealed by our previous study in addition to the reported cyclin-dependent kinase 4 (CDK4) selective inhibition. To uncover more details of this pharmacologically prospective property, this study further investigated whether fascaplysin had direct anti-proliferation effects on human umbilical vein endothelial cells (HUVEC), which might be contributing to anti-angiogenesis. The results showed that G1 cell cycle arrest was induced by 2.6 µM fascaplysin in a time-dependent manner, and exhibited more sensitive than hepatocarcinoma cells BeL-7402 and Hela cells. Approximately 56.09 ± 2.63% of the cells were arrested at the G1 phase after 24h, and 64.94 ± 2.07% after 36 h, comparing to the 22.82 ± 1.2% in methanol treated cells. Apoptosis of HUVEC cells was induced by 1.3 µM fascaplysin and indicated by the sub-G1, Hoechst staining, terminal deoxynucleotidyl transferase dUTP-mediated nicked end labeling (TUNEL) assay, and annexin-V and propidium (PI) label. This apoptosis response was further confirmed by the detection of active caspase-3 and by western blotting using antibodies against Bax, Bcl-2, procaspase-8, and Bid, indicating that apoptosis in HUVEC cells may involve a mitochondria pathway, by the demonstration of an increase in the Bax/Bcl-2 ratio. Together, our results suggest that the anti-angiogenesis activity of fascaplysin is through the direct effects of cell cycle arrest and apoptosis on HUVEC.

Design, synthesis, and primary activity evaluation of pyrrolidine derivatives as matrix metalloproteinase inhibitors.

A series of novel pyrrolidine derivatives was designed, synthesized, and assayed to determine the derivatives' activity against matrix metalloproteinase-2 (MMP-2) and aminopeptidase N (APN)/CD13. Preliminary biological tests showed that most compounds inhibit MMP-2 in a highly selective manner compared to APN. Compounds 9d, 9e, and 9g had better inhibitory activity than LY52 and could be used as lead compounds in the future.

QSAR studies of histone deacetylase (HDAC) inhibitors by CoMFA, CoMSIA, and molecular docking.

In order to develop highly potent antitumor agents, three-dimensional quantitative structure-activity relationship (3-D QSAR) studies were conducted using a series of thienyl-based hydroxamic acids. Comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) methods were applied to provide the structural information for further chemical modification and optimization. ClogP was applied as an additional descriptor in the CoMFA analysis to study the effects of lipophilic parameters on the activity of these compounds, and it did improve the statistical significance of the model. Two molecules were designed based on the 3-D QSAR analysis, their activity values were predicted by the generated model, and their binding mode was elucidated by a docking approach compared to molecules in the dataset.

A microplate-based screening assay for neuraminidase inhibitors.

Neuraminidase (NA) represents a highly promising new target for drug development in influenza virus genes. Rapid screening of enzyme inhibitors is a key method for the identification of leading compounds. In order to speed up the screening for enzyme inhibitors of natural and synthetic origin, effective and fast assays are needed. 2'-(4-Methylumbelliferyl)-α-D-N-acetylneuraminic acid (4-MUNANA) was selected as substrate for development of a microplate-based assay. The enzymatic reaction conditions were optimized as follows: in a 100 µL reaction mixture, the final concentrations were 32.5 mM sodium acetate (pH 3.5), 20 µM 4-MUNANA, 0.005% (w/v) bovine serum albumin, and 0.42 µg/mL NA. In the study, the doseresponse relationship of oseltamivir carboxylate to NA activity was observed. In addition, an overall Z' value of 0.8 proved the systems robustness and potential for screening. The assay system developed will be a valuable tool to discover new structures for the therapeutic inhibition of NA used to treat Influenza.

Novel N-hydroxybenzamide histone deacetylase inhibitors as potential anti-cancer agents.

Histone deacetylases (HDACs) are a class of Zn(2+) dependent metalloproteases that play an important role in tumorigenesis. Inhibition of HDACs may be a potential strategy for cancer therapy. This study designed and synthesized a series of novel N-hydroxybenzamide histone deacetylase inhibitors based on the structural features of suberoylanilide hydroxamic acid (SAHA), the first HDAC inhibitor that came to market. Preliminary biological evaluation in vitro found that most of the inhibitors had satisfactory inhibitory activity (IC(50) =1-17 µM) against HDACs and HCT116 tumor cells.