An improved approach to generate IL-15+/+/TGFßR2-/- iPSC-derived natural killer cells using TALEN.

We present a TALEN-based workflow to generate and maintain dual-edited (IL-15+/+/TGFßR2-/-) iPSCs that produce enhanced iPSC-derived natural killer (iNK) cells for cancer immunotherapy. It involves using a cell lineage promoter for knocking in (KI) gene(s) to minimize the potential effects of expression of any exogenous genes on iPSCs. As a proof-of-principle, we KI IL-15 under the endogenous B2M promoter and show that it results in high expression of the sIL-15 in iNK cells but minimal expression in iPSCs. Furthermore, given that it is known that knockout (KO) of TGFßR2 in immune cells can enhance resistance to the suppressive TGF-ß signaling in the tumor microenvironment, we develop a customized medium containing Nodal that can maintain the pluripotency of iPSCs with TGFßR2 KO, enabling banking of these iPSC clones. Ultimately, we show that the dual-edited IL-15+/+/TGFßR2-/- iPSCs can be efficiently differentiated into NK cells that show enhanced autonomous growth and are resistant to the suppressive TGF-ß signaling.

Actively Targeting Redox-Responsive Multifunctional Micelles for Synergistic Chemotherapy of Cancer.

Stimuli-responsive polymeric micelles decorated with cancer biomarkers represent an optimal choice for drug delivery applications due to their ability to enhance therapeutic efficacy while mitigating adverse side effects. Accordingly, we synthesized a digoxin-modified novel multifunctional redox-responsive disulfide-linked poly(ethylene glycol-b-poly(lactic-co-glycolic acid) copolymer (Bi(Dig-PEG-PLGA)-S2) for the targeted and controlled release of doxorubicin (DOX) in cancer cells. Within the micellar aggregate, the disulfide bond confers redox responsiveness, while the presence of the digoxin moiety acts as a targeting agent and chemosensitizer for DOX. Upon self-assembly in aqueous solution, Bi(Dig-PEG-PLGA)-S2 formed uniformly distributed spherical micelles with a hydrodynamic diameter (D h ) of 58.36 ± 0.78 nm and a zeta potential of -24.71 ± 1.01 mV. The micelles exhibited desirable serum and colloidal stability with a substantial drug loading capacity (DLC) of 6.26% and an encapsulation efficiency (EE) of 83.23%. In addition, the release of DOX demonstrated the redox-responsive behavior of the micelles, with approximately 89.41 ± 6.09 and 79.64 ± 6.68% of DOX diffusing from DOX@Bi(Dig-PEG-PLGA)-S2 in the presence of 10 mM GSH and 0.1 mM H2O2, respectively, over 96 h. Therefore, in HeLa cell lines, DOX@Bi(Dig-PEG-PLGA)-S2 showed enhanced intracellular accumulation and subsequent apoptotic effects, attributed to the targeting ability and chemosensitization potential of digoxin. Hence, these findings underscore the promising characteristics of Bi(Dig-PEG-PLGA)-S2 as a multifunctional drug delivery vehicle for cancer treatment.

Doxorubicin-loaded Polymeric Biotin-PEG-SeSe-PBLA Micelles with surface Binding of Biotin-Mediated Cancer Cell Targeting and Redox-Responsive Drug release for enhanced anticancer efficacy.

Biotin receptors are overexpressed in various cancer cell types, essential in tumor development, metabolism, and metastasis. Chemotherapeutic agents may be more effective and have fewer adverse effects if they specifically target the biotin receptors on cancer cells. Polymeric micelles (PMs) with nanoscale size via the EPR effect to accumulate near tumor tissue. We utilized the solvent exchange technique to crate polymeric Biotin-PEG-SeSe-PBLA micelles. This underwent self-assembly to create uniformly dispersed PMs with a hydrodynamic diameter of 81.54 ± 0.23 nm. The resulting PMs characterized by 1HNMR, 13CNMR, FTIR, and Raman spectroscopy. PMs exhibited a high efficacy of Doxorubicin encapsulation (EE) and loading content (DLC), with values of 5.93 wt% and 74.32 %, respectively. DOX@Biotin-PEG-SeSe-PBLA micelles showed optimal DOX release, around 89 % and 74 % in 10 mM glutathione and 0.1 % H2O2, respectively, within 72 hours, in the simulated cancer redox pool. Fascinatingly, the blank Biotin-PEG-SeSe-PBLA micelles did not affect the HaCaT or HeLa cell lines; approximately 85 % of the cells were metabolically active. Contrarily, at a 5 µg/ml concentration, DOX@Biotin-PEG-SeSe-PBLA specifically inhibited the proliferation of roughly 76 % of HeLa cells and 11 % of HaCaT cells. The fluorescence microscopy results demonstrated that biotin-decorated micelles were more successfully internalized by HeLa cells, which overexpress the biotin receptor, than by non-targeted micelles in vitro. In summary, the diselenide-linked Biotin-PEGSeSe-PBLA formed smart PMs that could offer DOX specific to cancer cells with precision and are physiologically durable.

Focusing of surface plasmon polaritons propagating at the SiO2/Ag interface with 2-level and 4-level Fresnel phase zone pad structures.

We propose and demonstrate dielectric Fresnel phase zone pad (FPZP) structures for focusing surface plasmon polaritons (SPPs) propagating at the SiO2/Ag interfaces. We exploited up-conversion fluorescence microscopy to characterize the SPP focusing. We first report on the SPP focusing with 2-level FPZP structures that introduced a π-phase shift in the SPP wavefront between adjacent zones. We optimized the SPP focusing by fine-tuning the longitudinal width of the FPZP structure. This led to the enhancement of the peak intensity of the SPP focal spot and the reduction of the focal spot size in both the longitudinal and transverse directions. Such focusing was also demonstrated with different focal lengths. To further improve the SPP focusing, we developed a 4-level FPZP structure, which introduced a π/2-phase shift in the SPP wavefront between adjacent zones. With the optimized 4-level FPZP structure, the SPP focal spot peak intensity is further improved, and the spot size is reduced. To assist the design of the FPZP structures, we carried out theoretical analysis and numerical calculations to determine the SPP wavelengths at various oxide/Ag interfaces. We also carried out finite difference time domain (FDTD) calculations to simulate the SPP focusing with the FPZP structures. The results of the FDTD simulation agree with the experimental results qualitatively.

The impact of COVID-19 epidemic on emergency department visits of older patients in Taiwan.

BACKGROUND: The number of emergency department (ED) visits has significantly declined since the COVID-19 pandemic. In Taiwan, an aged society, it is unknown whether older adults are accessing emergency care during the COVID-19 epidemic. Therefore, this study aimed to investigate the impact of COVID-19 on the ED visits and triage, admission, and intensive care unit (ICU) hospitalization of the geriatric population in a COVID-19-dedicated medical center throughout various periods of the epidemic. METHODS: A retrospective chart review of ED medical records from April 9 to August 31, 2021 were conducted, and demographic information was obtained from the hospital's computer database. The period was divided into pre-, early-, peak-, late-, and post-epidemic stages. For statistical analysis, one-way analysis of variance followed by multiple comparison tests (Bonferroni correction) were used. RESULTS: A statistically significant decrease in the total number of patients attending the ED was noted during the peak-, late-, and post-epidemic stages. In the post-epidemic stage, the number of older patients visiting ED was nearly to that of the pre-epidemic stage, indicating that older adults tend to seek care at the ED earlier than the general population. Throughout the entire epidemic period, there was no statistically significant reduction in the number of the triage 1& 2 patients seeking medical attention at the emergency department. In the entire duration of the epidemic, there was no observed reduction in the admission of elderly patients to our hospital or ICU through the ED. However, a statistically significant decrease was observed in the admission of the general population during the peak epidemic stage. CONCLUSIONS: During the peak of COVID-19 outbreak, the number of ED visits was significantly affected. However, it is noteworthy that as the epidemic was gradually controlled, the older patients resumed their ED visits earlier that the general population as indicated by the surge in their number. Additionally, in the patient group of triage 1& 2, which represents a true emergency, the number did not show a drastic change.

Association of Coracoacromial Ligament Degeneration With Rotator Cuff Tear Patterns and Retear Rate.

Background: Coracoacromial ligament (CAL) degeneration is thought to be a factor in external impingement in bursal-sided rotator cuff tears, but CAL release is associated with adverse effects. Purpose: To investigate the association between CAL degeneration and the patterns of massive rotator cuff tears using multiple modalities and to assess the effect of CAL degeneration on supraspinatus tendon retear rates. Study Design: Cohort study; Level of evidence, 2. Methods: The authors prospectively recruited 44 patients who had undergone arthroscopic rotator cuff repair without acromioplasty or CAL release. Preoperative radiographs and magnetic resonance imaging (MRI) scans were reviewed to determine acromial morphology and CAL thickness, respectively. Rotator cuff tears were categorized as isolated supraspinatus or massive (involvement of ≥2 tendons), with massive tears categorized using the Collin classification. Acromial degeneration was analyzed using the Copeland-Levy classification. The CAL was biopsied intraoperatively and histologically analyzed using the Bonar score. At 6-month follow-up, the integrity of the repaired supraspinatus tendon was analyzed on MRI using the Sugaya classification. Finally, the associations among CAL degeneration, rotator cuff tear pattern, and arthroscopic grading were investigated. Results: Patients with Collin type B rotator cuff tear had significantly higher CAL Bonar scores than those with Collin type A or isolated supraspinatus tears (10.0 vs 6.8 and 3.4; P = .03 and P < .001, respectively). Patients with a degenerative acromial undersurface of Copeland-Levy stage 2 or 3 had CALs with significantly higher Bonar scores than those with an intact acromial undersurface (8.4 and 8.2 vs 3.5; P = .034 and P = .027, respectively). The CAL Bonar scores of patients with different stages of the 6-month postoperative Sugaya classification were comparable (6.5, 7.2, 8.0, and 7.8 for stages 1, 2, 3, and 4, respectively; P = .751). Conclusion: CAL degeneration was more severe in anterosuperior-type massive rotator cuff tears. Interestingly, even without acromioplasty, the severity of CAL degeneration did not affect the retear rate of the supraspinatus tendon.

Derivation and Preclinical Characterization of CYT-303, a Novel NKp46-NK Cell Engager Targeting GPC3.

Glypican-3 (GPC3) is an oncofetal antigen that is highly expressed in multiple solid tumors, including hepatocellular carcinoma, and is barely expressed in adult normal tissues except the placenta. NKp46 activation receptor is expressed in all-natural killer (NK) cells, including tumor-infiltrating NK cells. FLEX-NKTM is a platform for the production of tetravalent multifunctional antibody NK cell engagers (NKE). CYT-303 was designed using the FLEX-NK scaffold, incorporating a novel humanized NKp46 binder that does not induce NKp46 internalization and a humanized GPC3 binder that targets the membrane-proximal lobe to mediate NK cell-redirected killing of HCC tumors. CYT-303 shows sub-nanomolar binding affinities to both GPC3 and NKp46. CYT-303 was highly potent and effective in mediating NK cell-redirected cytotoxicity against multiple HCC tumor cell lines and tumor spheroids. More interestingly, it can reverse the dysfunction induced in NK cells following repeated rounds of serial killing of tumors. It also mediated antibody-dependent cellular phagocytosis (ADCP) and complement-dependent cytotoxicity against GPC3-expressing HCC tumors. In vivo, CYT-303 showed no toxicity or cytokine release in cynomolgus monkeys up to the highest dose (60 mg/kg), administered weekly by intravenous infusion for 28 days. These results demonstrate the potential of CYT-303 to be a safe and effective therapy against HCC.

Poly(vinylidene fluoride) Intestinal Sleeve Implants for the Treatment of Obesity and Type 2 Diabetes.

Currently, treatment of diabetes and associated obesity involves Roux-en-Y gastric bypass or sleeve gastrectomy to reduce the absorption of nutrients from the intestine to achieve blood glucose control. However, the surgical procedure and subsequent recovery are physically and psychologically burdensome for patients, with possible side effects, so alternative treatments are being developed. In this study, two methods, solution casting and machine direction orientation (MDO), were used to prepare intestinal implants made of poly(vinylidene fluoride) (PVDF) film and implant them into the duodenum of type 2 diabetic rats for the treatment of obesity and blood glucose control. The PVDF film obtained by the MDO process was characterized by FT-IR, Raman spectroscopy, XRD and piezoelectricity tests, which showed higher composition of ß crystalline phase and better elongation and mechanical strength in specific directions. Therefore, the material was finally tested on rats after it was proven to be non-toxic by biological toxicity tests. The PVDF was implanted into alloxan-induced diabetic rats, which were used as a model of impaired insulin secretion due to pancreatic beta cell destruction rather than obesity-induced diabetes, and rats were tracked for 24 days, showing significantly improved body weight and blood glucose levels. As an alternative therapeutic option, intestinal sleeve implant showed future potential for application.

Post-Cholecystectomy Right Hepatic Artery Pseudoaneurysm Induced by Generalized Seizure: A Case Report.

Pseudoaneurysm is a rare complication of laparoscopic cholecystectomy (LC). In most cases, the patient presents with gastrointestinal bleeding or hemoperitoneum. Here, we present a case with a post-cholecystectomy right hepatic artery pseudoaneurysm (PSA) induced by a generalized seizure. A 39-year-old male was sent to the emergency room with a generalized seizure and a loss of consciousness for approximately 5 min. Diffuse abdominal pain was complained of after consciousness returned. The surgical history of LC 13 days prior was mentioned. Abdominal computer tomography (CT) revealed a lobulated fluid accumulation in the gallbladder fossa with prominent fatty stranding and suspected biloma formation. After admission for one week, sharp abdominal pain was observed. Abdominal CT angiography revealed a right hepatic artery pseudoaneurysm. Transcatheter arterial embolization was performed with a total of seven platinum coils. In conclusion, it is important for doctors to take pseudoaneurysm into consideration in the patient who presents with seizure attack after receiving LC. Late discovery of PSA when it is ruptured can lead to fatal conditions, such as severe hemoperitoneum.

Association Between Medial Displacement of the Middle Glenohumeral Ligament and Subscapularis Tear Severity.

Background: The intraoperative invisible middle glenohumeral ligament (MGHL) test has been shown to be associated with a subscapularis tendon retraction. However, the preoperative location of the MGHL and its association with subscapularis tear severity has not been evaluated. Purpose: To determine (1) the interrater reliability for identification and position of the MGHL, (2) any association between the MGHL position and subscapularis tears, and (3) the cutoff point at which MGHL position can predict subscapularis tear severity. Study Design: Cohort study (diagnosis); Level of evidence, 2. Methods: The magnetic resonance imaging (MRI) scans and surgical records of 176 patients were retrospectively reviewed by 3 independent orthopaedists. MGHL's identification, level (its position on axial MRI), and medial retraction ratio (distance from the lesser tuberosity to the MGHL divided by the width of the glenoid) were documented, and the interobserver agreement of the 3 indices was assessed. We calculated the association between subscapularis tears and the MGHL level and medial retraction ratio. Receiver operating characteristic (ROC) curve analyses were conducted to establish the optimal threshold of the MGHL medial retraction ratio to predict subscapularis tear. Results: The MGHL was identified by at least 2 reviewers in 124 individuals (70.5%). The interobserver reliability was very good for MGHL identification (κ = 0.766), moderate for MGHL level (κ = 0.582), and excellent for MGHL medial retraction ratio (intraclass correlation coefficient = 0.848). A low, positive correlation between MGHL level and subscapularis tear severity was found (Somers d = 0.392, P < .001), as well as a significant association between the medial retraction ratio and Lafosse classification of subscapularis tear size (P < .001). A medial retraction ratio of ≥1.25 had a sensitivity of 0.70, a specificity of 0.83, and a positive likelihood ratio of 4.20, with excellent accuracy (area under the ROC curve = 0.820) to predict severe subscapularis tear. Conclusion: The MGHL was identified in 70.5% of shoulder MRIs. The location of the MGHL on preoperative MRI, as described by its level and the medial retraction ratio, was significantly associated with subscapularis tear severity, and a medial retraction ratio of ≥1.25 was predictive of a severe subscapularis tear.

Comparison of Biomechanical Failure Loads Between Tape-Type and Conventional Sutures in Internal Knotless Anchor-Based Constructs.

Background: Despite the increasing prevalence of tape-type sutures, whether internal knotless anchors can consistently affix tape-type sutures has not been thoroughly investigated. Purpose: To evaluate whether substituting tape-type sutures for conventional sutures influences the suture-holding strength of internal knotless anchors. Study Design: Controlled laboratory study. Level of evidence, 5. Methods: A total of 3 internal knotless anchors were tested: a spiral core clamping anchor (Footprint Ultra PK), a winged clamping anchor (PopLok), and a spooling anchor (ReelX STT). Four constructs were compared for each type of anchor, with the anchor double or quadruple loaded with tape-type sutures or conventional sutures. The testing protocol comprised preloading suture tension to 10 N; cyclic loading, in which tension increased in increments of 10 N from 10 to 90 N; and a load-to-failure stage set at a speed of 0.5 mm/s. The clinical failure load (CFL) was defined as suture slippage of ≥3 mm. Also, 1-way analysis of variance and power analysis were used to compare the CFLs of the constructs. Results: For the quadruple-loaded spiral core clamping anchors, a significant reduction in CFLs was seen with conventional sutures over tape-type sutures (138.10 ± 4.73 vs 80.00 ± 12.25 N, respectively; P < .001). This reduction was not observed under the double-loaded condition (conventional vs tape type: 76.00 ± 5.48 vs 80.00 ± 10.00 N, respectively). Substitution of the suture materials did not significantly reduce the CFLs for the winged clamping anchors (conventional vs tape type: 40.00 ± 10.00 vs 30.00 ± 7.07 N for double loaded, respectively, and 64.00 ± 13.41 vs 50.00 ± 10.00 N for quadruple loaded, respectively) or the spooling anchors (conventional vs tape type: 62.00 ± 19.23 vs 56.32 ± 20.20N for double loaded, respectively, and 72.00 ± 21.68 vs 84.00 ± 13.42 N for quadruple loaded, respectively). Conclusion: Substituting tape-type sutures for conventional sutures increased the CFLs of some internal knotless anchors. With specific suture-anchor combinations, quadruple-loaded conventional suture anchors had CFLs higher than those of double-loaded conventional suture anchors. Clinical Relevance: When multiple tape-type sutures are used in conjunction with a clamping anchor, clinicians should note a possible reduction in CFLs and resultant early suture slippage.

Does Weaver-Dunn procedure have a role in chronic acromioclavicular dislocations? A meta-analysis.

BACKGROUND: In treatment of chronic acromioclavicular (AC) joint dislocations, both the Weaver-Dunn procedure (WD) and CC ligament reconstruction (CCR) are recommended options due to the low possibility of healing of the coracoclavicular (CC) ligaments. The aim of this review was to determine whether CCR will yield favorable clinical and radiographic outcomes in the treatment of chronic AC dislocations. METHOD: The Cochrane Library, EMBASE, and PubMed databases were searched for literature on chronic AC dislocations from data inception to June 30, 2021. Patient data were pooled using standard meta-analytic approaches. The Cochrane-Mantel-Haenszel method and variance-weighted means were used to analyze the outcomes. The Review Manager version 5.3 software (The Nordic Cochrane Centre, The Cochrane Collaboration, Copenhagen, Denmark) was used to calculate the heterogenicity, mean difference, and relative risk (RR) for all outcomes in the meta-analysis. RESULTS: The current analysis included four trials on this topic, and all AC joint dislocations were classified as Rockwood types III to VI. The pooled data showed that the CCR group had significantly better post-operative American Shoulder and Elbow Surgeons Shoulder (ASES) scores, Oxford Shoulder Scores (OSSs), and Nottingham Clavicle Scores (NCSs) than the WD group, with a significant difference (p < 0.001, p = 0.020, and p < 0.001, respectively). In terms of the post-operative Constant-Murley Scores (CMSs), there were no significant differences between the CCR group and the WD group (p = 0.100). The CCR group had significantly better post-operative abduction and flexion of the index shoulder than the WD group (p < 0.001 and p < 0.001, respectively). In terms of radiological outcomes, the post-operative coracoclavicular distance (CCD) with a 10 kg load was smaller in the CCR group compared to that in the WD group (p < 0.001). The overall surgical wound infection rate was 11.6% in the WD group and 12.9% in the CCR, respectively (p = 0.82). CONCLUSION: The CCR group had better clinical outcome scores in the ASES, OOS, NCS, abduction, flexion, and external rotation than the WD group. In terms of radiological outcomes, the CCR group showed less displacement in weight-loaded post-CCD than the WD group, which indicated that the CCR provided more stability and resistance to deformation forces.

Biomechanical comparisons of hook plate and screw fixations in split-type greater tuberosity fractures of the humerus.

BACKGROUND: Screws and plate are commonly utilized for the fixation of split-type humeral greater tuberosity (GT) fractures. However, the mechanical properties of these 2 types of fixation methods have not been compared directly. The aim of the present study was to evaluate the classic 2 screws fixation with hook locking plate from a mechanical perspective. METHODS: Sixteen synthetic humerii (Sawbones Pacific Research Laboratories, Vashon, WA, USA) were divided into 2 groups. An osteotomy was performed to simulate a split-type GT fracture. Group A (n = 8) was fixed with 2 standard parallel screws. Group B (n = 8) was fixed with a hook plate. Each specimen was tested with traction force at 45° shoulder abduction. Following the 20-N preload, a 500-cycle loading test was applied with a force ranging from 20 to 200 N (valley/peak), and the interfragmental displacement was measured periodically at intervals of 100 cycles. Finally, all the specimens were pulled with destructive force at a rate of 5 mm/min until catastrophic failure. RESULTS: The hook plate exhibited greater construct stiffness than the screw fixation (63.2 ± 6.1 N/mm vs. 40.9 ± 3.4 N/mm, P < .001). All of the specimens completed the entire cyclic loading test without catastrophic failure, and the fragment displacement after 500 cycles was 0.4 ± 0.2 mm for the hook plate and 2.1 ± 0.3 mm for screw fixation, which was statistically lower in the plate group (P < .001). In terms of failure load, the hook plate group exhibited a significantly greater value than the screw group (770.6 ± 94.6 vs. 427.5 ± 45.1 N/mm, P < .001). The failure modes of both fixation methods were distinct. CONCLUSION: In humeral GT fracture fixation, hook plate fixation appears to offer greater construct stiffness and failure load while maintaining fragment stability to resist a physiological traction force. The current study provides support from a mechanical perspective for the clinical application of the hook plate.

Owen, Sugaya, and Hayashida Classifications Give Poor Intra- and Inter-Rater Agreement on a Magnetic Resonance Imaging Evaluation of Subscapularis Tendon Retears.

PURPOSE: To investigate the intra- and inter-rater agreements for magnetic resonance imaging (MRI) evaluations of subscapularis tendon integrity at 6 months after arthroscopic rotator cuff repairs. METHODS: Patients who had isolated or combined subscapularis tears and had undergone arthroscopic rotator cuff repairs were retrospectively included. The exclusion criteria included revision of arthroscopic surgery, minor subscapularis tears without repair, and inadequate postoperative images. MRI scans 6 months after surgery were used for the purpose of accessing the integrity of the subscapularis tendons. Three orthopaedic surgeons blindly evaluated the images twice at 2-week intervals. Three currently available classifications were used: the Owen classification, the Sugaya classification, and the Hayashida classification. Dichotomization and trichotomization methods were used for the Sugaya classification and Hayashida classifications. The aforementioned classification scores were combined for the agreement evaluation. Intra- and inter-rater agreement was assessed by calculating the Fleiss' kappa coefficients. RESULTS: A total of 35 patients were included. Both the Owen and Hayashida classifications had poor inter-rater agreements (κ = 0.10 and 0.04, respectively) and poor-to-weak intra-rater agreements (κ = 0.27-0.44 and 0.16-0.45, respectively). The Sugaya classification had poor inter-rater agreement (κ = 0.10) and poor intra-rater agreements (κ = 0.16-0.32). Dichotomization and trichotomization of Sugaya and Hayashida classifications did not lead to superior agreements. The classification combination resulted in poor inter- and intra-rater agreements (κ = 0.01-0.12 and 0.08-0.39, respectively). CONCLUSIONS: The Owen classification, Sugaya classification, and Hayashida classification had poor intra- and inter-rater agreement in terms of evaluating subscapularis tendon re-tears on 6 months' postoperative MRI. The dichotomized and trichotomized classifications as well as the combined classifications from currently available classifications did not lead to superior agreements. LEVEL OF EVIDENCE: Level IV, diagnostic: case series.

Targeted HAI-2 deletion causes excessive proteolysis with prolonged active prostasin and depletion of HAI-1 monomer in intestinal but not epidermal epithelial cells.

Mutations of SPINT2, the gene encoding the integral membrane, Kunitz-type serine inhibitor HAI-2, primarily affect the intestine, while sparing many other HAI-2-expressing tissues, causing sodium loss in patients with syndromic congenital sodium diarrhea. The membrane-bound serine protease prostasin was previously identified as a HAI-2 target protease in intestinal tissues but not in the skin. In both tissues, the highly related inhibitor HAI-1 is, however, the default inhibitor for prostasin and the type 2 transmembrane serine protease matriptase. This cell-type selective functional linkage may contribute to the organ-selective damage associated with SPINT 2 mutations. To this end, the impact of HAI-2 deletion on matriptase and prostasin proteolysis was, here, compared using Caco-2 human colorectal adenocarcinoma cells and HaCaT human keratinocytes. Greatly enhanced prostasin proteolytic activity with a prolonged half-life and significant depletion of HAI-1 monomer were observed with HAI-2 loss in Caco-2 cells but not HaCaT cells. The constitutive, high level prostasin zymogen activation observed in Caco-2 cells, but not in HaCaT cells, also contributes to the excessive prostasin proteolytic activity caused by HAI-2 loss. HAI-2 deletion also caused increased matriptase zymogen activation, likely as an indirect result of increased prostasin proteolysis. This increase in activated matriptase, however, only had a negligible role in depletion of HAI-1 monomer. Our study suggests that the constitutive, high level of prostasin zymogen activation and the cell-type selective functional relationship between HAI-2 and prostasin renders Caco-2 cells more susceptible than HaCaT cells to the loss of HAI-2, causing a severe imbalance favoring prostasin proteolysis.

Long-term oncologic result of laparoscopic versus open gastrectomy for gastric cancer: a propensity score matching analysis.

BACKGROUND: Laparoscopic gastrectomy is an acceptable procedure for early-stage gastric cancer; however, most patients are diagnosed at an advanced stage and older age in Taiwan. The feasibility and safety of applying laparoscopic gastrectomy in daily practice remain unclear. This study aimed to examine the short- and long-term outcomes of laparoscopic gastrectomy versus open procedures. METHODS: From 2007 to 2015, 192 patients who underwent open gastrectomy and 189 patients who underwent laparoscopic gastrectomy for gastric cancer at a single center were included. Propensity score matching analysis was used to adjust selection biases associated with age, preoperative hemoglobin, the extent of resection, tumor size, and stage of the disease. The demographics, perioperative parameters, short-term postoperative results, and 5-year survival data were analyzed. RESULTS: Open gastrectomy was more frequently performed in the elderly, larger tumor size, advanced stage of the disease, and disease requiring total gastrectomy or combined organ resection. After propensity score matching, 108 patients with laparoscopic gastrectomy were compared to 108 patients with open gastrectomy. The morbidity rates were not different in both groups (25.9%), while hospital stay was shorter in the laparoscopic group (16.0 vs. 18.8 days, p = 0.04). The 5-year overall survival and disease-free survival were superior in the laparoscopic group (p = 0.03 and p = 0.01, respectively); however, the survival differences were not significant in the subgroup analysis by stage. Laparoscopic gastrectomy had fewer recurrences than open gastrectomy. The pattern of recurrence was not different between the groups. CONCLUSIONS: Laparoscopic gastrectomy can be safely applied in both early and locally advanced gastric cancer without compromising oncologic outcomes. TRIAL REGISTRATION: Retrospective registration.

Outcomes of Infection and Risk of Mortality in Liver Transplant Patients with Simultaneous Splenectomy.

PURPOSE: The purpose of this study was to compare the outcomes of infection between liver transplant patients with and without simultaneous splenectomy. METHODS: We retrospectively analyzed the records of 211 patients who underwent liver transplantation in the Tri-Service General Hospital from 2012 to 2017. The frequency of blood cultures obtained after liver transplantation; incidence of bacteremia, pathogens, and complications; and overall survival rates were compared between the groups. RESULTS: One hundred thirty-three of 211 patients underwent liver transplantation without simultaneous splenectomy. There were no significant differences in the frequency of blood cultures obtained after liver transplantation (non-splenectomy group and splenectomy group: 63% and 62%, respectively); incidences of bacteremia after liver transplantation (21% and 21%, respectively), repeat bacteremia (39% and 35%, respectively), cytomegalovirus infection (4% and 3%, respectively), herpes infection (6% and 7%, respectively), and fungal infection (3% and 3%, respectively); and overall survival rate between the two groups. However, there was a significant difference in infection-related deaths between the groups. Simultaneous splenectomy and episodes of antibody-related rejection were significant risk factors associated with infection-related death in multivariate analyses. CONCLUSION: Although simultaneous splenectomy does not increase the incidence of infection, simultaneous splenectomy definitely carries risks of infection-related mortality in liver transplantation.

Wound drainage after proximal femoral nail antirotation (PFNA) fixation may negatively affect the patients with intertrochanteric fractures: A prospective randomized controlled trial.

INTRODUCTION: The effect of using closed suction drainage system with the proximal femoral nail antirotation (PFNA) system fixation on outcomes in treating intertrochanteric fractures (ITFs) is still unknown. This prospective randomized controlled trial aimed to examine whether routine drainage is useful for PFNA fixation in ITFs. METHODS: A total of 80 patients with acute ITFs were treated with closed or mini-open reduction with PFNA fixation at the National Cheng Kung University Hospital and 60 eligible patients were randomized for whether to receive suction drainage. In clinical outcomes, the visual analog scale (VAS), morphine equivalent dosage, injured thigh width, body temperature, wound condition and wound infection were measured on postoperative day 1, 2, 4, 10, and 90. In laboratory outcomes, we evaluated hemoglobin and hematocrit levels postoperatively at different time points. Blood transfusion and total blood loss (TBL) were measured by Mercuriali's formula in millimeter. RESULTS: The results revealed that the amount of blood transfusion received by the drained group (543.3 mL) was more than that by the undrained group (367.8 mL; p = 0.0074), and similarly, TBL in the drained group (750.1 mL) was more than that in the undrained group (537.4 mL; p = 0.0067). Regarding clinical and laboratory outcomes, compared with the undrained group, the drained group had a higher VAS score on postoperative day 2 (p = 0.0216). No difference was observed between the 2 groups for morphine equivalent dosage, thigh swelling, wound infection and hematoma, hospitalization period, or total number of complications at every time point after index procedure. CONCLUSIONS: Blood transfusion requirement and TBL were higher in the drained group than in the undrained group of PFNA fixation for ITFs. In addition, the closed drainage system may have manifested no short-term benefit for wound condition postoperatively.

TRIM71 binds to IMP1 and is capable of positive and negative regulation of target RNAs.

TRIM71 is an important RNA-binding protein in development and disease, yet its direct targets have not been investigated globally. Here we describe a number of disease and developmentally-relevant TRIM71 RNA targets such as the MBNL family, LIN28B, MDM2, and TCF7L2. We describe a new role for TRIM71 as capable of positive or negative RNA regulation depending on the RNA target. We found that TRIM71 co-precipitated with IMP1 which could explain its multiple mechanisms of RNA regulation, as IMP1 is typically thought to stabilize RNAs. Deletion of the NHL domain of TRIM71 impacted its ability to bind to RNA and RNAs bound by congenital hydrocephalus-associated point mutations in the RNA-binding NHL domain of TRIM71 clustered closely with RNAs bound by the NHL deletion mutant. Our work expands the possible mechanisms by which TRIM71 may regulate RNAs and elucidates further potential RNA targets.