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The oncogenic fusion protein promyelocytic leukemia/retinoic acid receptor alpha (PML/RARα) is critical for acute promyelocytic leukemia (APL). PML/RARα initiates APL by blocking the differentiation and increasing the self-renewal of leukemic cells. The standard clinical therapies all-trans retinoic acid (ATRA) and arsenic trioxide (ATO), which induce PML/RARα proteolysis, have dramatically improved the prognosis of APL patients. However, the emergence of mutations conferring resistance to ATRA and ATO has created challenges in the treatment of APL patients. Exploring pathways that modulate the oncogenic activity of PML/RARα could help develop novel therapeutic strategies for APL, particularly for drug-resistant APL. Herein, we demonstrated for the first time that palmitoylation of PML/RARα was a critical determinant of its oncogenic activity. PML/RARα palmitoylation was found to be catalyzed mainly by the palmitoyltransferase ZDHHC3. Mechanistically, ZDHHC3-mediated palmitoylation regulated the oncogenic transcriptional activity of PML/RARα and APL pathogenesis. The knockdown or overexpression of ZDHHC3 had respective effects on the expression of proliferation- and differentiation-related genes. Consistently, the depletion or inhibition of ZDHHC3 could significantly arrest the malignant progression of APL, particularly drug-resistant APL, whereas ZDHHC3 overexpression appeared to have a promoting effect on the malignant progression of APL. Thus, our study not only reveals palmitoylation as a novel regulatory mechanism that modulates PML/RARα oncogenic activity but also identifies ZDHHC3 as a potential therapeutic target for APL, including drug-resistant APL., (© 2024. The Author(s), under exclusive licence to Shanghai Institute of Materia Medica, Chinese Academy of Sciences and Chinese Pharmacological Society.)

Background: It has been argued that the use of artificial turf football fields in the National Football League (NFL) increases player injury risk compared with natural grass surfaces., Purpose/hypothesis: The purpose of this study was to quantify the rate of lower extremity injuries occurring in NFL players on artificial turf compared with natural grass surfaces and characterize the time missed due to injury and proportion of injuries requiring surgery. It was hypothesized that lower extremity injuries requiring surgical intervention would occur at a higher rate on artificial turf than on natural grass., Study Design: Descriptive epidemiology study., Methods: Lower extremity injury data for the 2021 and 2022 NFL seasons were obtained using publicly available records. Data collected included injury type, player position, player age, playing surface, weeks missed due to injury, and whether the patient underwent season-ending or minor surgery. Multivariable logistic regression was performed to determine the risk of season-ending surgery according to playing surface., Results: When combining injuries for the 2021 and 2022 seasons (N = 718 injuries), the incidence rate of lower extremity injury was 1.22 injuries/game for natural grass and 1.42 injuries/game for artificial turf. The odds of a season-ending surgery were found to be significantly higher on artificial turf compared with natural grass (odds ratio = 1.60; 95% CI, 1.28-1.99; P < .05), while additional variables, including weather, age, position, week of injury occurrence, and history of prior injury, did not influence the odds of season-ending surgery., Conclusion: The 2021 and 2022 NFL seasons of our analysis demonstrated a higher incidence rate of injuries on artificial turf surfaces compared with natural grass surfaces. In addition, the odds of injury requiring season-ending surgery were found to be significantly higher on artificial turf compared with natural grass., Competing Interests: One or more of the authors has declared the following potential conflict of interest or source of funding: N.K.P. has received education payments from Evolution Surgical. B.T.F. has received education payments from Evolution Surgical. AOSSM checks author disclosures against the Open Payments Database (OPD). AOSSM has not conducted an independent investigation on the OPD and disclaims any liability or responsibility relating thereto. Ethical approval was not sought for the present study., (© The Author(s) 2024.)

Background Context: Multilevel cervical myelopathy is a common cause of spinal cord dysfunction in adults. Surgical intervention via laminoplasty can provide satisfactory clinical outcomes by expansive decompression of the spinal cord. Traditional suture or bone graft techniques have been associated with insufficient fixation, leading to premature closure and subsequent neurological deterioration. In contrast, plated laminoplasty has been shown to provide stable fixation to maintain canal enlargement, but longer-term outcomes are lacking., Purpose: To evaluate longer-term clinical outcomes and reoperations associated with plate-only open-door laminoplasty., Study Design: Retrospective review of prospectively collected data., Patient Sample: Postoperative patients who underwent plate-only open door laminoplasty with minimum 5-year follow up., Outcome Measures: modified Japanese Orthopaedic Association (mJOA) score, Neck Disability Index (NDI), and 12-item Short Form Health Survey (SF-12)., Methods: All patients at a single academic institution who underwent plate-only open-door cervical laminoplasty from 9/1/2006 to 9/1/2016 were identified to ensure minimum 5 year follow up. Clinical outcomes included the modified Japanese Orthopaedic Association (mJOA) score, the Neck Disability Index (NDI), and the 12-item Short Form Health Survey (SF-12). The occurrence of any repeat operations on the cervical spine was evaluated, as well as its cause. The study team attempted to contact all eligible patients to achieve at least 5 years postoperative follow-up. Pairwise t tests were performed to compare clinical outcomes at preoperative, 6 months, 1-year, and final postoperative follow-up with an α level of 0.05., Results: A total of 774 met the initial inclusion criteria, of which 157 were included in the study (20.3%). Most common reasons for exclusion included inability to reach after 3 attempts (49.48%), inactive phone numbers (20.28%), and patient declining (3.49%). Included patients had an average age of 60.66±10.63 and an average follow-up time of 8.37±2.57 years (minimum 5 years). mJOA scores (preoperative 11.59±2.16) improved significantly at 6-months (14.57±2.07, p<.001), 1-year (15.19±1.95, p<.001), and final follow-up (14.59±2.63, p<.001). NDI (preoperative 33.89±18.54) improved significantly at 6 months (27.89±19.72, p=.03), 1-year (25.96±19.79, p=.01) and final follow-up (17.88±17.17, p<.001). SF-12 MCS (preoperative 44.73) improved significantly at 6 months (52.01, p=.001), 1-year (51.62, p=.008), and final follow-up (52.32, p<.001). No patient underwent reoperations for plate failure or canal closure with recurrent stenosis. Reoperations for progressive spondylosis during the follow up period were rare and occurred in only three patients for new onset radiculopathy (1.9%) and two patients for myelopathy (1.3%) at an average of 3.2 years postoperative. There were no reoperations performed for adjacent segment disease., Conclusions: At a minimum of 5 years and an average of more than 8 years postoperative, laminoplasty was associated with significant and sustained improvements in mJOA, NDI, and SF-12 MCS. Importantly, no patients underwent revision surgery for plate failure or recurrent canal closure. Reoperations for new onset radiculopathy and myelopathy were also very rare over the 8-year average follow-up period, with no reoperations for adjacent segment disease. Plate-only laminoplasty is a durable means of treating multilevel myelopathy with excellent longer-term outcomes and a very low risk of reoperation, either for premature closure or the inevitable spondylotic changes that occur over time in patients with similar baseline characteristics to the study population., Competing Interests: Declaration of Competing Interest One or more of the authors declare financial or professional relationships on ICMJE-TSJ disclosure forms., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Background: Bronchopulmonary dysplasia (BPD) is characterized by an arrest in lung development and is a leading cause of morbidity in premature neonates. It has been well documented that BPD disproportionally affects males compared to females, but the molecular mechanisms behind this sex-dependent bias remain unclear. Female mice show greater preservation of alveolarization and angiogenesis when exposed to hyperoxia, accompanied by increased miR-30a expression. In this investigation, we tested the hypothesis that loss of miR-30a would result in male and female mice experiencing similar impairments in alveolarization and angiogenesis under hyperoxic conditions., Methods: Wild-type and miR-30a -/- neonatal mice were exposed to hyperoxia [95% FiO 2 , postnatal day [PND1-5] or room air before being euthanized on PND21. Alveolarization, pulmonary microvascular development, differences in lung transcriptome, and miR-30a expression were assessed in lungs from WT and miR-30a -/- mice of either sex. Blood transcriptomic signatures from preterm newborns (with and without BPD) were correlated with WT and miR-30a -/- male and female lung transcriptome data., Results: Significantly, the sex-specific differences observed in WT mice were abrogated in the miR-30a -/- mice upon exposure to hyperoxia. The loss of miR-30a expression eliminated the protective effect in females, suggesting that miR-30a plays an essential role in regulating alveolarization and angiogenesis. Transcriptome analysis by whole lung RNA-Seq revealed a significant response in the miR-30a -/- female hyperoxia-exposed lung, with enrichment of pathways related to cell cycle and neuroactive ligand-receptor interaction. Gene expression signature in the miR-30a -/- female lung associated with human BPD blood transcriptomes. Finally, we showed the spatial localization of miR-30a transcripts in the bronchiolar epithelium., Conclusions: miR-30a could be one of the biological factors mediating the resilience of the female preterm lung to neonatal hyperoxic lung injury. A better understanding of the effects of miR-30a on pulmonary angiogenesis and alveolarization may lead to novel therapeutics for treating BPD., (© 2023. Society for Women's Health Research and BioMed Central Ltd.)

Hypothesis: The purpose of this study was to analyze the SHR of patients diagnosed with small (SRCTs) and massive rotator cuff tears (MRCTs), adhesive capsulitis (AC), and glenohumeral osteoarthritis (GH-OA) and compare their measurements to those of patient controls with healthy shoulders using DDR. We hypothesize that various diagnoses will vary with regards to SHR., Methods: The sequences of pulsed radiographs collated in DDR to create a moving image were prospectively analyzed during humeral abduction in normal controls and in 4 distinct shoulder pathology groups: SRCT, MRCT, AC, and GH-OA. GH and ST joint angles were measured at 0°-30°, 30°-60°, 60°-90°, and maximal coronal plane humeral abduction. SHR was defined as the ratio of the change in humeral abduction over the change in scapula upward rotation during humeral abduction and was calculated within the above angle intervals., Results: A total of 121 shoulders were analyzed. Forty normal controls were compared to 13 SRCTs, 29 MRCTs, 16 AC, and 23 GH-OA. SHR during humeral abduction differed significantly in patients with MRCT (1.91 ± 0.72), AC (1.55 ± 0.37), and GH-OA (2.31 ± 1.01) compared to controls (3.39 ± 0.79). When analyzed across 30° intervals of abduction, there was a significantly lower SHR found at 0°-30°, 30°-60°, and 60°-90° in MRCT, AC, and GH-OA across each motion range compared to controls. Control patients had an arc of abduction of 103° ± 32°, which was significantly larger than all other pathologies (MRCT: 76° ± 23°, SRCT: 81° ± 21°, AC: 65° ± 27°, GH-OA: 71° ± 35°) and an average scapular abduction of 33° ± 14°, which was significantly less than patients with an MRCT (46° ± 10°) and AC (65° ± 27°)., Conclusion: SHR remained significantly lower throughout shoulder abduction in MRCT (43.65%), AC (-54.29%), and GH-OA (32.01%) compared to controls. When isolating for humeral and scapular motion, all 4 pathologies had decreased GH abduction, whereas AC and MRCT had an increased scapular compensatory motion compared to controls. Quantifying kinematic patterns like SHR using DDR can be implemented as a novel, safe, and cost-effective method to diagnose shoulder pathology and to monitor response to treatment., (Copyright © 2023 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier Inc. All rights reserved.)

Background: A posterior urethral diverticulum (PUD) is a serious postoperative complication after anorectal malformation correction. Complete resection is technical demanding because of limited retrourethral working space deep in the pelvis., Objective: We pioneered the single-incision laparoscopic approach for PUD excision and evaluated the efficacy., Design, Setting, and Participants: Twenty-six PUD patients undergoing redo surgeries between June 2011 and June 2021 were reviewed., Surgical Procedure: A series of transabdominal retraction sutures were placed through the PUD to facilitate dissection. The contents were evacuated to create a working space. Distal PUD dissection was carried along the submucosal layer to prevent injury of the urethra/pelvic nerve complex. The rectal mucosa was peeled off from the junction site for complete PUD excision. The muscular cuff of the distal rectum was then oversewn., Measurements: Operative time, postoperative recovery, and complications were assessed., Results and Limitations: The mean age of redo surgery was 2.46 yr. The average operative duration was 2.35 h. The mean postoperative hospital stay, resumption of full diet, and bowel movement were 10.23, 2.15, and 1.54 d, respectively. The median follow-up period was 46 mo (12-132 mo). No remnant of PUD, recurrent fistula, or urinary leak was detected. None of the patients had difficulty in urination, urinary dribbling, urinary tract infection, constipation, or soiling. All patients retained morning erection, and two postpubertal patients had ejaculations., Conclusions: Our single-incision laparoscopic redo surgery provides an effective approach for PUD excision. It minimizes complications. It also preserves urinary and bowel continence and sexual function., Patient Summary: Complete resection of a posterior urethral diverticulum (PUD) in anorectal malformation is technically demanding because of limited retrourethral working space in the deep pelvis. The outcomes of single-incision laparoscopic PUD excision were satisfactory., (Copyright © 2022 European Association of Urology. Published by Elsevier B.V. All rights reserved.)

Behavioral observations suggest a connection between anxiety and predator defense, but the underlying neural mechanisms remain unclear. Here we examine the role of the anterior hypothalamic nucleus (AHN), a node in the predator defense network, in anxiety-like behaviors. By in vivo recordings in male mice, we find that activity of AHN GABAergic (AHN Vgat+ ) neurons shows individually stable increases when animals approach unfamiliar objects in an open field (OF) or when they explore the open-arm of an elevated plus-maze (EPM). Moreover, object-evoked AHN activity overlap with predator cue responses and correlate with the object and open-arm avoidance. Crucially, exploration-triggered optogenetic inhibition of AHN Vgat+ neurons reduces object and open-arm avoidance. Furthermore, retrograde viral tracing identifies the ventral subiculum (vSub) of the hippocampal formation as a significant input to AHN Vgat+ neurons in driving avoidance behaviors in anxiogenic situations. Thus, convergent activation of AHN Vgat+ neurons serves as a shared mechanism between anxiety and predator defense to promote behavioral avoidance., (© 2022. The Author(s).)

Distal radius fractures (DRFs) are among the most common fractures in the United States. Despite their high incidence, there is no consensus on the optimal type of cast or splint to treat these fractures. The purpose of this systematic review is to evaluate the available literature pertaining to the outcomes for different constructs used to conservatively treat DRFs. A literature search of PubMed, Medline, and Embase was conducted to identify research comparing the outcomes of various immobilization mechanisms. In particular, endpoints included complications (eg, loss of reduction, pain), radiographic outcomes, and Disabilities of the Arm, Shoulder, and Hand (DASH) scores. A total of 1655 articles were identified during the literature search, and 22 ultimately fulfilled inclusion criteria. These 22 studies included 1826 conservatively treated DRFs. The different immobilization mechanisms were divided into 8 groupings: above-elbow casts, above-elbow splints, below-elbow casts, below-elbow splints, gutter or spica casts, gutter or spica splints, dorsal-volar splints, and dorsal splints. Qualitative review of the studies determined that below-elbow constructs result in equal or better functional and radiologic outcomes when compared with above-elbow constructs. Meta-analysis demonstrated that a statistically significant difference ( P = .04) existed in the incidence of loss of reduction between the immobilization constructs, although post hoc analysis did not detect significant differences between 2 particular constructs.

Background: The timing of anorectoplasty for anorectal malformations (ARMs) is controversial and essential for long-term bowel function. The advantages of laparoscopy make primary anoplasty possible for neonates. This study aims to report the medium-term results of the one-stage laparoscopic-assisted anorectoplasty (LAARP) and compare them with the staged LAARP., Methods: This study included 242 boys who underwent LAARP between June 2013 and December 2018 in our center. Forty-five neonatal patients successfully underwent the one-stage LAARP, and the remaining 197 patients who had already undergone colostomy received staged procedures. The complications and bowel function were compared between the two groups after 1:1 propensity score matching (PSM)., Results: Before matching, age at assessment and classification were significantly different between the two groups (P < 0.05). After PSM, 42 patients were included in each of the two groups, and the patients were well balanced. The overall occurrence of postoperative complications in the one-stage group was significantly lower than in the staged group (P < 0.05). The median follow-up periods were 55.0 and 54.5 months, respectively. The mean value of BFS was similar in the two groups. After medical management, a second evaluation was conducted to distinguish overflow pseudo-incontinence from patients with no bowel control potential. The functional outcome in the two groups is comparable (P = 0.307)., Conclusions: One-stage LAARP is safe and feasible for neonates with high-and intermediate-type ARMs. Compared with staged LAARP, one-stage LAARP has fewer complications and comparable functional outcomes. The authors recommend these procedures to be performed in dedicated centers with sufficient expertise., Level of Evidence: Level III., Competing Interests: Declaration of Competing Interest All authors have no conflict of interest to disclose., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Objective: To evaluate fertility clinic management of male factor infertility, including website educational content as well as factors associated with referral for urologic evaluation and care., Materials and Methods: Using 2015-2018 Centers for Disease Control and Prevention Fertility Clinic Success Rates Reports, 480 operative fertility clinics in the United States were identified. Clinic websites were systematically reviewed for content regarding male infertility. Structured telephone interviews of clinic representatives were performed to determine clinic-specific practices for management of male factor infertility. Multivariable logistic regression models were used to predict how clinic characteristics (geographic region, practice size, practice setting, proximity to urologist, in-state andrology fellowship, state-mandated fertility coverage, annual in vitro fertilization cycles, and percentage of in vitro fertilization cycles for male factor infertility) were associated with patient referral to a urologist for male infertility care., Results: We interviewed 477 fertility clinics and analyzed available websites (n = 474). The majority of websites (77%) discussed male infertility evaluation while 46% discussed treatment. Fifty clinics (11%) had an on-site urologist. Clinics with on-site urologists were more likely to be larger practices, academically affiliated, and discuss male infertility treatment on their website (all P ≤ .05). For clinics without an on-site urologist, practice size and presence of an in-state andrology fellowship program were the strongest predictors of urologic referral (P <.02)., Conclusion: Variability in patient-facing education and infertility practice setting and size influence access to urologic care for couples with male factor infertility., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Introduction: We evaluated fertility clinic management of male factor infertility, including patient education and referral for urological evaluation and care., Methods: Using 2015-2018 Centers for Disease Control and Prevention Fertility Clinic Success Rates Reports, 480 operative fertility clinics in the United States were identified. Clinic websites were systematically reviewed for content regarding male infertility. Structured telephone interviews of clinic representatives were performed to determine clinic-specific practices for management of male factor infertility. Multivariable logistic regression models were used to predict how clinic characteristics (geographic region, practice size, practice setting, in-state andrology fellowship, state-mandated fertility coverage, annual in vitro fertilization cycles and percentage of in vitro fertilization cycles for male factor infertility) were associated with reproductive endocrinologist physician management of male infertility and/or referral to a urologist., Results: We interviewed 477 fertility clinics and analyzed available websites (474). The majority of websites (77%) discussed male infertility evaluation, while 46% discussed treatment. Clinics that were academically affiliated, had an accredited embryo laboratory and referred patients to a urologist were less likely to have the reproductive endocrinologist manage male infertility (all p <0.05). Practice affiliation, practice size and website discussion of surgical sperm retrieval were the strongest predictors of nearby urological referral (all p <0.05)., Conclusions: Variability in patient-facing education, and fertility clinic setting and size influence fertility clinics' management of male factor infertility.

Purpose: We evaluated the literature on complications associated with different positions used for immobilizing the upper extremity during conservative treatment of distal radius fractures (DRF)., Methods: A search of PubMed, Embase, and Medline was conducted to identify original research on the effects that upper extremity positioning during the treatment of DRFs has on complication rates. Treatment groups were categorized by wrist positioning in flexion, extension, or neutral, as well as forearm positioning in pronation, supination, or neutral. The primary endpoints examined included the loss of reduction, recasting/refabricating an orthosis, and functional limitations., Results: A total of 1,655 articles were identified through an initial database search. Ultimately, 8 studies, with 786 total patients, met the inclusion criteria for this systematic review. A qualitative analysis determined that immobilizing DRFs with the wrist in extension results in better functional and radiographic outcomes with lower rates of complications, such as pain, recasting, and the need for operation. The 2 studies that compared forearm pronation versus supination revealed contradictory results regarding which position was associated with superior outcomes. A meta-analysis comparing the various wrist and forearm positions failed to demonstrate statistically significant differences in the rates of loss of reduction or recasting/refabricating an orthosis between the groups. This analysis was limited by considerable heterogeneity in the data from the different studies., Conclusions: Despite the high incidence of DRFs, there is limited research on the optimal position of immobilization for conservative treatment of them. Available evidence suggests that the wrist should be immobilized in extension, as these patients had improved functional and radiographic outcomes. No conclusion can be drawn from the existing literature on ideal forearm position during immobilization. This review also suggests better data reporting practices for studies researching DRFs, so that future meta-analyses can be more comprehensive., Type of Study/level of Evidence: Therapeutic II., (© 2021 The Authors.)

Background: For the last 20 years, laparoscopy management of anorectal malformations (ARM) has been challenged due to the development of postoperative urethral diverticulum or injury caused by the imprecise transection of rectourethral fistulae, particularly rectobulbar fistulae situated deep in the pelvis. We have developed a combined approach of enteroscopy and laparoscopy for intraluminal incision of a rectourethral fistula., Methods: We retrospectively reviewed 47 ARM patients who underwent surgical corrections using the combined approach between January 2019 and June 2020. Early postoperative and subsequent follow-up results were evaluated., Results: The median follow-up period was 12 months. The average age at surgery was 3.18 ± 0.64 months. The mean operative time of a single-incision laparoscopic-assisted anorectoplasty (SILAARP) was 1.19 ± 0.29 h. The time for intraluminal incision of the fistula was shortened from 14 to 2 min. No patients underwent a conversion. The average postoperative hospital stay, time to full feeds and placement of an anal tube were 10 days, 1 day, and 5 days, respectively. No urethral diverticulum, urinary injury, wound infection, rectal retraction, anal stenosis or rectal prolapse was encountered in the cohort., Conclusions: The combined enteroscopy and laparoscopy approach offers precise management of rectourethral fistulae. It could effectively obviate urethral complications, eliminating the obstacles of laparoscopy application in the management of ARMs.

Purpose: We sought to review the clinical outcomes of conservative and operative treatment options for acute distal radioulnar joint (DRUJ) instability associated with distal radius fractures in adult patients., Methods: A systematic search of PubMed, MEDLINE, and EMBASE for articles published between 1990 and 2020 involving DRUJ instability associated with distal radius fractures was performed. The primary outcomes analyzed included clinical grip strength; range of motion; the disability of the arm, shoulder and hand (DASH) score; and the modified Mayo wrist score (MMWS)., Results: Of the 531 articles identified in the literature search, 8 met our defined criteria and were included in the final analysis. The cumulative sample size was 258 patients at a mean follow-up of 11.1 months (range, 3-16.9 months). Treatment groups included cast immobilization in supination, K-wire stabilization, and triangular fibrocartilage complex (TFCC) repair. Statistical analysis revealed no difference across groups in active flexion-extension or DASH scores. A significant decrease in grip strength was found in patients who underwent TFCC repair compared with that in those who underwent both cast immobilization ( P  = .04) and K-wire stabilization ( P  = .02). Furthermore, we found a significant decrease in active pronation-supination between patients who underwent TFCC repair and those who underwent cast immobilization ( P  = .03). Patients who underwent TFCC repair were also found to exhibit decreased MMWS as compared with those who underwent K-wire stabilization ( P  = .05). Overall, persistent DRUJ instability was only found in 4 patients (1.5%), without a significant difference between treatment groups., Conclusions: This study suggests functional advantages of certain treatment modalities over others, with the range of motion being highest in patients who underwent cast immobilization and grip strength being highest in patients who underwent K-wire stabilization. However, the mean DASH scores showed no difference across all groups, calling into question the clinical need to pursue operative treatment via K-wire stabilization or TFCC repair over conservative treatment via cast immobilization. This study will hopefully serve as a foundation for future prospective studies to help improve and standardize treatment algorithms in patients with DRUJ instability and distal radius fractures. Type of study/level of evidence : Therapeutic II., (© 2021 The Authors.)

The hypothalamus regulates innate social interactions, but how hypothalamic neurons transduce sex-related sensory signals emitted by conspecifics to trigger appropriate behaviors remains unclear. Here, we addressed this issue by identifying specific hypothalamic neurons required for sensing conspecific male cues relevant to inter-male aggression. By in vivo recording of neuronal activities in behaving mice, we showed that neurons expressing dopamine transporter (DAT + ) in the ventral premammillary nucleus (PMv) of the hypothalamus responded to male urine cues in a vomeronasal organ (VNO)-dependent manner in naive males. Retrograde trans-synaptic tracing further revealed a specific group of neurons in the bed nucleus of the stria terminalis (BNST) that convey male-relevant signals from VNO to PMv. Inhibition of PMv DAT+ neurons abolished the preference for male urine cues and reduced inter-male attacks, while activation of these neurons promoted urine marking and aggression. Thus, PMv DAT+ neurons exemplify a hypothalamic node that transforms sex-related chemo-signals into recognition and behaviors., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)

AGRP (agouti-related neuropeptide) expressing inhibitory neurons sense caloric needs of an animal to coordinate homeostatic feeding. Recent evidence suggests that AGRP neurons also suppress competing actions and motivations to mediate adaptive behavioral selection during starvation. Here, in adult mice of both sexes we show that AGRP neurons form inhibitory synapses onto ∼30% neurons in the medial preoptic area (mPOA), a region critical for maternal care. Remarkably, optogenetically stimulating AGRP neurons decreases maternal nest-building while minimally affecting pup retrieval, partly recapitulating suppression of maternal behaviors during food restriction. In parallel, optogenetically stimulating AGRP projections to the mPOA or to the paraventricular nucleus of hypothalamus but not to the LHA (lateral hypothalamus area) similarly decreases maternal nest-building. Chemogenetic inhibition of mPOA neurons that express Vgat (vesicular GABA transporter), the population targeted by AGRP terminals, also decreases maternal nest-building. In comparison, chemogenetic inhibition of neurons in the LHA that express vesicular glutamate transporter 2, another hypothalamic neuronal population critical for feeding and innate drives, is ineffective. Importantly, nest-building during low temperature thermal challenge is not affected by optogenetic stimulation of AGRP→mPOA projections. Finally, via optogenetic activation and inhibition we show that distinctive subsets of mPOA Vgat+ neurons likely underlie pup retrieval and maternal nest-building. Together, these results show that AGRP neurons can modulate maternal nest-building, in part through direct projections to the mPOA. This study corroborates other recent discoveries and underscores the broad functions that AGRP neurons play in antagonizing rivalry motivations to modulate behavioral outputs during hunger. SIGNIFICANCE STATEMENT In order for animals to initiate ethologically appropriate behaviors, they must typically decide between behavioral repertoires driven by multiple and often conflicting internal states. How neural pathways underlying individual behaviors interact to coherently modulate behavioral outputs, in particular to achieve a proper balance between behaviors that serve immediate individual needs versus those that benefit the propagation of the species, remains poorly understood. Here, by investigating projections from a neuronal population known to drive hunger behaviors to a brain region critical for maternal care, we show that activation of AGRP→mPOA projections in females dramatically inhibits maternal nest-building while leaving mostly intact pup retrieval behavior. Our findings shed new light on neural organization of behaviors and neural mechanisms that coordinate behavioral selection., (Copyright © 2019 the authors 0270-6474/19/390456-16$15.00/0.)

Objectives: The present meta-analysis aimed to evaluate the short- and long-term outcomes of laparoscopic surgery (LS) versus open surgery (OS) for rectal cancer., Methods: PubMed, Web of Science, and Cochrane Library, were searched for eligible randomized controlled trials (RCTs) published up to June 2017. Operation related index, postoperative complication, and long-term survival rate and disease-free survival rate were evaluated by meta-analytical techniques., Result: Nine RCTs enrolling 4126 patients were included in the present meta-analysis. Compared to OS, LS had similar positive circumferential resection margin (CRM) and number of lymph nodes extracted (LNE) as well as long term 5 years survival rate and disease-free survival rate, but of which the risk tendency was higher in LS group. The short-term outcomes of major and total postoperative complication were lower in LS group., Conclusions: LS for rectal cancer was as safe and effective as OS in terms of long-term outcomes, but with lower postoperative complication.

The visual system is remarkably efficient at extracting summary statistics from the environment. Yet at any given time, the environment consists of many groups of objects distributed over space. Thus, the challenge for the visual system is to summarize over multiple groups. The current study investigates the capacity and computational efficiency of ensemble perception, in the context of perceiving mean sizes of multiple spatially intermixed groups of circles. In a series of experiments, participants viewed an array of one to eight sets of circles. Each set contained four circles in the same colors, but with different sizes. Participants estimated the mean size of a probed set. The set that would be probed was either known before onset of the array (pre-cue condition) or afterwards (post-cue condition). By comparing estimation error in the pre-cue and post-cue conditions, we found that participants could reliably estimate mean sizes for approximately two sets (Experiment 1). Importantly, this capacity was robust against attention bias toward individual objects in the sets (Experiment 2). Varying the exposure time to stimulus arrays did not increase the capacity limit, suggesting that ensemble perception could be limited by an internal resource constraint, rather than the speed of information encoding (Experiment 3). Moreover, we found that the visual system could not encode and hold more individual items than ensemble representations (Experiment 4). Taken together, these results suggest that ensemble perception provides an efficient way of information processing but with constraints.

Background: Adolescents with chronic health conditions and/or disabilities (CHC/D's) often face challenges when transitioning to adult care, which leads to a higher risk of morbidity and mortality. Although it is recommended that establishing the medical home and family physician (FP) attachment prior to transfer will improve health outcomes, there is little evidence or policy surrounding the role of the FP during this transition. This study explores the described use of health services by adolescents with CHC/D's, as well as the adolescent's and caregiver's perceptions of their FP., Methods: Participants were recruited from the British Columbia Children's Hospital, Canada and a multi-method phased approach was used. In Phase 1, 84 adolescent and caregiver pairs completed questionnaires asking what medical services they accessed for specific health needs. In Phase 2, another cohort of 21 adolescent and caregiver pairs completed the same questionnaires and were interviewed regarding their perception of their FP., Results: 96% (n = 81) of adolescents with CHC/D's in phase 1 had a FP. Thirty four percent (n = 34) of adolescents had not seen their FP in the last 6 months. While adolescents with CHC/D mostly accessed their FP for primary health issues, they frequently also accessed specialists for prescription refills (50%, n = 51), mental health (29%, n = 30) and sexual health (16%, n = 16). While most adolescent/caregiver participants reported positive communication and trust in their FP, some had a poor understanding of the FP's role in coordinating care., Conclusion: As many adolescents with CHC/D may see their FP infrequently and may not clearly understand their role, opportunities exist for strengthening primary care home attachment as well as adolescent and caregiver literacy around the potential contribution of the FP during and after transfer to adult services. Responsibility for improving care coordination for this population should be ideally shared between FPs and pediatric specialists.

Finite element simulation of facial mimics provides objective indicators about soft tissue functions for improving diagnosis, treatment and follow-up of facial disorders. There is a lack of in vivo experimental data for model development and validation. In this study, the contribution of the paired Zygomaticus Major (ZM) muscle contraction on the facial mimics was investigated using in vivo experimental data derived from MRI. Maximal relative differences of 7.7% and 37% were noted between MRI-based measurements and numerical outcomes for ZM and skin deformation behaviors respectively. This study opens a new direction to simulate facial mimics with in vivo data.

Background and Purpose: Icariin is a potent stimulator of osteogenic differentiation; however, the mechanism underlying its osteogenic effect remains unclear. The osteogenic effect of icariin is related to the upstream glucocorticoid-induced leucine zipper (GILZ) signaling pathway, and antagonism with dexamethasone-induced osteoblast inhibition was noted., Methods: MC3T3-E1 cells were cultured in induced medium treated with icariin with or without dexamethasone. After short interfering RNA (siRNA) were used to silence GILZ expression, the degree of mineralization, proliferation, and GILZ expression as well as the levels of osteogenic (OPG, RANKL, ALP, OC and RUNX2) markers were tested., Results: Dexamethasone inhibited, while icariin increased, osteogenic activity, as indicated by ALP activity and calcium nodules. Meanwhile, dexamethasone dose-dependently (10 -6 M-10 -4 M) increased GILZ and RANKL expression and reduced ALP, OPG and OC, but the pattern of mRNA expression was reversed when icariin was added. Furthermore, GILZ (dexamethasone-induced) inhibition caused by icariin or moderately silenced by GILZ siRNA abolished the osteogenesis inhibition effect of dexamethasone, as indicated by the changes in the GILZ, ALP, OPG and RANKL expression levels; ALP activity; and calcium nodule., Conclusions: These results indicate that the GILZ-mediated osteogenic signal pathway is involved in the osteogenic effect induced by icariin., (Copyright © 2017 Elsevier Masson SAS. All rights reserved.)

ATP-binding cassette transporter A1 (ABCA1) mediates formation of disc-shaped high-density lipoprotein (HDL) from cell lipid and lipid-free apolipoprotein A-I (apo A-I). Discoidal HDL particles are heterogeneous in physicochemical characteristics for reasons that are understood incompletely. Discoidal lipoprotein particles similar in characteristics and heterogeneity to cell-formed discoidal HDL can be reconstituted from purified lipids and apo A-I by cell-free, physicochemical methods. The heterogeneity of reconstituted HDL (rHDL) is sensitive to the lipid composition of the starting lipid/apo A-I mixture. To determine whether the heterogeneity of cell-formed HDL is similarly sensitive to changes in cell lipids, we investigated four compounds that have well-established effects on cell lipid metabolism and ABCA1-mediated cell cholesterol efflux. 2-Bromopalmitate, D609, monensin and U18666A decreased formation of the larger-sized, but dramatically increased formation of the smaller-sized HDL. 2-Bromopalmitate did not appear to affect ABCA1 activity, subcellular localization or oligomerization, but induced dissolution of the cholesterol-phospholipid complexes in the plasma membrane. Arachidonic and linoleic acids shifted HDL formation to the smaller-sized species. Tangier disease mutations and inhibitors of ABCA1 activity wheat germ agglutinin and AG 490 reduced formation of both larger-sized and smaller-sized HDL. The effect of probucol was similar to the effect of 2-bromopalmitate. Taking rHDL formation as a paradigm, we propose that ABCA1 mutations and activity inhibitors reduce the amount of cell lipid available for HDL formation, and the compounds in the 2-bromopalmitate group and the polyunsaturated fatty acids change cell lipid composition from one that favors formation of the larger-sized HDL particles to one that favors formation of the smaller-sized species., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Purpose: The Rex shunt has been employed successfully to treat patients with extrahepatic portal hypertension. In the conventional Rex shunt, the internal jugular vein is used as a venous graft. Inevitably, such a procedure requires neck exploration and sacrifice of the internal jugular vein. The authors describe a novel adaptation of spleen-preserving spleno-Rex bypass, successfully carried out in children with extrahepatic portal hypertension., Methods: The mean age of the four patients (1 boy, 3 girls) was 46 months at the time of operation. All children had a history of upper gastrointestinal bleeding, and suffered from splenomegaly and hypersplenism. Spleen-preserving proximal splenic-left intrahepatic portal shunt was performed in all patients. The splenic artery and vein were ligated at the splenic hilum, and the splenic vein was completely separated from the bed of the pancreas to its junction with the inferior mesenteric vein. The freed splenic vein was anastomosed to left portal vein. The short gastric and left gastroepiploic vessels were kept intact to supply and drain the spleen. All patients were followed-up for 7-33 months (median: 21.5 months)., Results: The spleen-preserving spleno-Rex bypass was successfully performed in all 4 patients. The median operative time was 225 min (range: 215-260 min). One patient received blood transfusion, and the postoperative length of hospital stay varied from 4 to 6 days (median: 4.5 days). Intraoperative portal venous angiography demonstrated the patency of the shunt in all patients. Postoperatively, the complete blood count normalized and the biochemistry tests were within normal range. Postoperative ultrasound confirmed shunt patency and satisfactory flow in the proximal splenic-portal shunt in each patient. The size of the spleen decreased and there was no recurrence of variceal bleeding., Conclusions: The spleen-preserving spleno-Rex bypass is a viable option to treat EHPVO in children., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Purpose: Single-incision laparoscopic surgery (SILS) as a surgical approach in treatment of pancreatic disease has recently been reported in adults. However, its application in pancreatic surgery in children is limited. In this article, we report our preliminary experience of SILS in children with pancreatic disease., Methods: Three children with pancreatic tumor underwent single-incision laparoscopic partial pancreatectomy between July 2011 and August 2013. Two of three children were girls, and one was a boy. The ages ranged from 2 to 10 months, with an average age of 6.7 months., Results: All operations were successfully performed. There was no conversion to the conventional multi-incision surgery. The mean operation time of the 3 cases was 153.3 minutes (range 120-200 minutes). The postoperative hospital stay was 7 days. The drainage tubes were kept for 3 to 4 days after surgery. There was no pancreatic juice leak in this case series. All patients were followed up and there was no recurrence., Conclusions: Single-incision laparoscopic partial pancreatectomy for children with pancreatic tumor is feasible., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Purpose: Hirschsprung-associated enterocolitis (HAEC) is a serious complication of Hirschsprung's disease (HD), with generalized sepsis and high mortality rate. Although the surgical correction of HD is mostly successful, challenges remain in the management of children with repeated episodes of enterocolitis. The authors describe a novel modification of transanal rectal mucosectomy and partial internal anal sphincterectomy (TRM-PIAS) for HD., Methods: One hundred twenty-seven HD children aged from 8 days to 16 years who successfully underwent TRM-PIAS were reviewed. TRM-PIAS was carried out circumferentially along the anorectal line. Anterior dissection was conducted between the rectal submuscosal layer and the rectal muscular sleeve. The posterior dissection was performed along the plane between internal and external anal sphincters. Normal colon was pulled through and anastomosed to anal mucosa. Aganglionic segment, rectal mucosa, part of internal anal sphincter and posterior rectal muscular cuff were removed. Twenty-five age-matched children without defecation dysfunction were used as the control group in the study of anal resting pressure., Results: Patients were followed up for 6-12 years (median: 8.2 years). The median age at last follow-up was 12.2 years (7.2-20.1 years). The incidence of enterocolitis decreased from 33.9% (43/127) preoperatively to 1.6% (2/127) postoperatively (P<0.01). The incidence of constipation decreased from 100% (127/127) preoperatively to 2.4% (3/127) postoperatively (P<0.01). Soiling rate on postoperative 1 month was 32.3%. It gradually decreased to 1.6% 6 months later. Anorectal manometries showed that mean anal resting pressure was significantly reduced from 37.9±12.5 mm Hg preoperatively to 20.2±6.4 mm Hg on postoperative 1 month and 24.8±9.9 mm Hg on postoperative 6 months, which were similar to age-matched normal controls (27.9±9.6 mm Hg, P>0.05)., Conclusions: TRM-PIAS is effective in treatment of HD. It is associated with low postoperative HD-associated enterocolitis., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Objective: To compare the effect of transumbilical single-site single-port with that of transumbilical single-site double-port laparoscopic varicocelectomy in the treatment of varicocele in adolescents., Methods: We randomly assigned 80 varicocele patients aged 10 - 16 years to two groups of equal number to receive transumbilical single-site single-port and single-site double-port laparoscopic varicocelectomy, respectively. We compared the operation time, postoperative hospital stay, incisional pain, complications and satisfaction with the abdominal cosmetic outcomes between the two groups., Results: All the operations were successfully performed. The double-port group showed a significantly higher score on the Visual Analogue Scale than the single-port group (4.8 +/- 1.4 vs 3.6 +/- 1.1, t = -4.986, P < 0.01), but there were no significant differences between the two groups in the operation time ([29.8 +/- 4.2] vs [31.2 +/- 4.6] min, t = 1.383, P = 0.171), postoperative hospital stay ([1.95 +/- 0.7] vs [1.82 +/- 0.8] d, t = -0.784, P = 0.436), complications (0 vs 0) and scores on the satisfaction with abdominal cosmetic outcomes (4.6 +/- 0.6 vs 4.8 +/- 0.5, t = 1.253, P = 0.214). No recurrence, umbilical hernia, hydrocele and orchiatrophy were found in the two groups of patients at 6 months after operation, and no visible scar was observed on the abdominal surface., Conclusion: With strict surgical indications, single-site single-port and single-site double-port laparoscopic varicocelectomies have similar clinical effects in the treatment of varicocele, which leave no scar on the abdominal surface. Single-site double-port laparoscopy needs no special instruments and therefore is worthier of wide clinical application.

Purpose: The aim of this study is to compare the long term outcomes between laparoscopic-assisted anorectoplasty (LAARP) and posterior sagittal anorectoplasty (PSARP) for children with rectobladderneck and rectoprostatic fistula anorectal malformations (ARM)., Methods: Thirty-two ARM children with rectobladderneck and rectoprostatic fistula who underwent LAARP between October 2001 and March 2012 were reviewed. The outcomes were compared with those of 34 ARM children who underwent PSARP between August 1992 and September 2001. The sacral ratio (SR), age at operation, operative time, postoperative hospital stay and complications were evaluated. Bowel functions were assessed using the Krickenbeck classification., Results: The mean operative time of the LAARP was significantly shorter than that of PSARP group (1.62 ± 0.40 vs 2.13 ± 0.30 h). The postoperative hospital stay was significantly shorter in the LAARP group (5.8 ± 0.65 vs 8.4 ± 0.67 h). The wound infections (11.8% vs 0%) and recurrent fistula (11.8% vs 0%) were more common in PSARP patients. Th e overall morbidity rate of PSARP group was significantly higher than that of the LAARP group (35.3% vs 12.5%, p<0.05). However, 7.5% of the LAARP patients developed rectal prolapse. Twenty-four of 32 patients were followed up for more than 3 years in LAARP group. The median follow up period was 7.5 years (range 4-11) in LAARP patients and 15.5 years (range 11-20) in PSARP patients. The rates of voluntary bowel movement, soiling (grade 1, 2 & 3) were similar in both groups. More patients from PSARP group developed grade 2 or 3 constipation (22.5% vs 0%, P<0.01)., Conclusions: Compared to PSARP, LAARP is a less invasive procedure. The long term functional outcomes after LAARP were equivalent if not better than those of PSARP., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Purpose: The application of laparoscopic surgery in pancreatic surgery in children is limited. In this article, we describe laparoscopic pancreaticojejunostomy for children with congenital pancreatic ductal dilatation., Methods: Four children with recurrent pancreatitis and pancreatic ductal dilatation underwent laparoscopic pancreaticojejunostomy between July 2009 and November 2011. Longitudinal incision of the dilated pancreatic ducts and side-to-side Roux-Y pancreaticojejunostomy were performed., Results: Operative time ranged from 103 to 154 min, and blood loss was minimal. The average postoperative hospital stay was 4 to 6 days. There were no pancreatic leaks. None of the patients experienced recurrence of pancreatitis., Conclusions: Laparoscopic pancreaticojejunostomy for children with congenital pancreatic ductal dilatation is safe and effective., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Purpose: Extrahepatic portal venous obstruction is the most common cause of portal hypertension in children. The Rex shunt has been used successfully to treat patients with extrahepatic portal hypertension. In the conventional Rex shunt, the internal jugular vein is used as a venous graft. Inevitably, such a procedure requires neck exploration and sacrifice of internal jugular vein. The authors describe a novel adaptation of gastroportal shunt, successfully carried out in 8 children with extrahepatic portal hypertension., Methods: The mean age of the 8 patients (6 boys and 2 girls) was 66.6 months at the time of operation. All children had portal hypertension. Seven had a history of upper gastrointestinal bleeding, and 4 had splenomegaly and hypersplenism. Gastroportal shunt was performed in all patients. The left gastric vein was mobilized and anastomosed to left portal vein. In 1 patient, the left gastric vein was not of adequate length and required a venous graft (the inferior mesenteric vein). All patients were followed up for 3 to 20 months (median, 9 months)., Results: The gastroportal shunt was successfully performed in all patients. The median operative time was 265 minutes (range, 205-360 minutes). Operative blood loss was 21 ± 7.4 mL, and the length of hospital stay varied from 9 to 19 days (median, 15 days). Intraoperative portal venous angiography demonstrated the patency of the shunt in all patients. Postoperatively, the complete blood count normalized, and the biochemistry tests were within reference range. Postoperative ultrasound confirmed shunt patency and satisfactory flow in the gastroportal shunt in each patient. The size of spleen decreased. There was no recurrence of variceal bleeding., Conclusions: The gastroportal shunt is an effective treatment of extrahepatic portal hypertension., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The diversity-oriented synthesis of dipeptide mimetic compounds embedded with bioactive molecular skeletons have been successfully established via microwave-assisted reactions between various 4-arylidene-2-phenyloxazol-5(4H)-ones and a broad scope of amines including aliphatic, aromatic, and heteroaromatic ones. This synthetic approach has prominent features of short reaction time, high yields, operational simplicity, as well as widespread applications, leading to a facile and straightforward access to structurally diverse dipeptide analogues with potential bioactivities. Moreover, the preliminary evaluation on the cytotoxic activity of this type of dipeptide derivatives has resulted in the finding of five compounds with stronger cytotoxicity than doxorubicin hydrochloride at the concentration of 10 μg/mL.

The efficient synthesis of novel 3-aminohexahydrocoumarin derivatives with high diastereoselectivity, wide applicability, short reaction time, high yields as well as operational simplicity was achieved via microwave-assisted reactions of dimedone or cyclohexane-1,3-dione with 4-arylidene-2-phenyloxazol-5(4H)-ones. Moreover, these novel compounds were subject to the test of in vitro cytotoxicity to carcinoma SW1116 and SGC7901 cells. Seven compounds showed stronger cytotoxicity to carcinoma SW1116 cells than doxorubicin hydrochloride at the concentration of 10 ug/mL. Nearly all the tested compounds exhibited strong or moderate cytotoxicity to SGC7901 cells with IC(50) values from 5.127 to 0.158 ug/mL., (Copyright © 2010. Published by Elsevier Masson SAS.)

This study was aimed at the transport across blood-brain barrier (BBB) of polysorbate-80 modified neurotoxin loaded polybutylcyanoacrylate nanoparticle (P-80-NT-NP) and its cytotoxicity. An in vitro model of BBB using rat brain microvascular endothelial cells (rBMECs) was established. The cytotoxicity of P-80-NT-NP was measured by the MTT assays, where neurotoxin (NT), nanoparticle (NP), neurotoxin nanoparticle (NT-NP) as control, and the permeability of P-80-NT-NP was determined by using of Millicell insert coculture with rBMECs and fluorescence spectrophotometry. MTT results showed that NT, NP, NT-NP and P-80-NT-NP were avirulent to rBMECs when the concentration of NT was lower than 200 ng x mL(-1). But the cytotoxicity of NP, NT-NP and P-80-NT-NP would be augmented accordingly as concentration increased (P < 0.01), causing obvious reductions of cell survival rate, with no significant difference between them (P > 0.05). When the concentration of NT was 150 ng x mL(-1), the permeability on rBMECs of P-80-NT-NP and NT-NP were both significantly higher than that of NT (P < 0.01), and the permeability of P-80-NT-NP was greater than that of NT-NP (P < 0.05). In conclusion, polysorbate-80 modified neurotoxin nanoparticles can transport across the BBB, while concentration of NT is greater than 200 ng x mL(-1), P-80-NT-NP has a little cytotoxicity against rBMECs.

Objective: To identify the Mycobacterium tuberculosis complex by detecting the secretory protein MPT64., Methods: The gene mpt64 was amplified by polymerase chain reaction (PCR) from the genome of Mycobacterium tuberculosis H(37)Rv strain and cloned into expression vector. Immune sera from rabbits by recombinant proteins MPT64, were used to make enzyme-labeled antibodies and coated antibodies. A double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was established to detect the secretory protein MPT64 of the culture supernatants in Mycobacterium strains. Results of ELISA were compared to those of the gene mpt64 amplified by PCR., Results: A recombinant vector was constructed. The minimum detectable concentration of MPT64 was 0.01 mg/L. A total of 27 reference strains and 170 clinical isolate strains were evaluated. PCR for Mycobacterium tuberculosis reference strain, Mycobacterium bovis reference strain and Mycobacterium africanum reference strain was positive, but that for other reference stains was negative, consistent with the results of ELISA. In the 170 clinical isolate stains, the positive result of PCR and ELISA was 98.2% (111/113) and 97.3% (110/113) respectively, while the specificity of PCR and ELISA was both 100%, no positive result in non-tuberculosis mycobacterium strains., Conclusion: Identification of the Mycobacterium tuberculosis complex by detecting secretory protein MPT64 is rapid, sensitive, and specific, which can be used routinely in clinical laboratories.

Objective: To establish a recombinant plasmid of CFP32 of Mycobacterium tuberculosis in E. coli, and to analyze its antigenicity., Methods: Rv0577 gene was amplified by polymerase chain reaction from genome of Mycobacterium tuberculosis, and then cloned into vector pMD18-T followed by the subclone into the expression vector pET21a. Recombinant CFP32 was expressed and purified. The antigenicity of the recombinant protein was analyzed by using Western-blot. The purified recombinant CFP32 protein was used as an antigen to screen the sera of 7 pulmonary TB patients (n = 97), as well as the other pulmonary disease patients (n = 25), and the clinically healthy controls (n = 38) by ELISA., Results: Recombinant plasmid of CFP32 was established, and be expressed efficiently in E. coli BL21 (DE3). The relative molecular mass of the protein was about 300,000 by SDS-PAGE analysis. The protein purified by Ni-NTA was in a purity over 90%, which was confirmed by Western-blot analysis. ELISA analysis showed its sensitivity and specificity were 63.9% (62/97) and 96.8% (2/63) respectively., Conclusion: The recombinant expression plasmid pET21a CFP32 has been constructed and CFP32 proteins has been successfully expressed and be purified in E. coli and, ELISA analysis has identified the recombinant CFP32 as a candidate antigen for TB serodiagnosis.

A pea actin isoform PEAc1 with green fluorescent protein (GFP) fusion to its C-terminus and His-tag to its N-terminus, was expressed in prokaryotic cells in soluble form, and highly purified with Ni-Chelating Sepharose Fast Flow column. The purified fusion protein (PEAc1-GFP) efficiently inhibited DNase I activities before polymerization, and activated the myosin Mg-ATPase activities after polymerization. The PEAc1-GFP also polymerized into green fluorescent filamentous structures with a critical concentration of 0.75 uM. These filamentous structures were labeled by TRITC-phalloidin, a specific agent for staining actin microfilaments, and identified as having 9 nm diameters by negative staining. These results indicated that PEAc1 preserved the essential characteristics of actin even with His-tag and GFP fusion, suggesting a promising potential to use GFP fusion protein in obtaining soluble plant actin isoform to analyze its physical and biochemical properties in vitro. The PEAc1-GFP was also expressed in tobacco BY2 cells, which offers a new pathway for further studying its distribution and function in vivo.