Stage IV gastric adenocarcinoma with enteroblastic differentiation with 5-year relapse-free survival after D2 gastrectomy and chemotherapy: A case report.

BACKGROUND: Gastric adenocarcinoma with enteroblastic differentiation (GACED), a rare subtype of gastric cancer, is associated with a more aggressive behavior than conventional gastric adenocarcinomas. We report a rare case of stage IV GACED treated with D2 gastrectomy and postoperative chemotherapy. CASE PRESENTATION: A 39-year-old woman with acute upper abdominal pain immediately underwent surgery for gastric perforation. Afterward she was diagnosed with adenocarcinoma of the pylorus. D2 gastrectomy was performed and the final pathological diagnosis was stage IV GACED with positive peritoneal cytology. Postoperative chemotherapy was initiated with S1 plus oxaliplatin for 1 year, which was ceased thereafter to enhance her quality of life. The patient survived more than 5 years without relapse after gastrectomy. CONCLUSIONS: Stage IV GACED, determined by positive spalt-like transcription factor 4, can be successfully treated with surgery and chemotherapy.

Simultaneous medial closing wedge distal femoral varus osteotomy and double-bundle anterior cruciate ligament reconstruction in the symptomatic femoral valgus deformity: A case report.

Distal femoral varus osteotomy (DFVO) is a widely recognized surgical procedure used to address valgus malalignment in patients with knee joint disorders. However, it still remains unclear whether anterior cruciate ligament (ACL) reconstruction can be performed in a single procedure along with DFVO. Herein, we present a 73-year-old female patient who developed lateral osteoarthritis of the knee with valgus alignment due to chronic ACL deficiency following a twisting injury during skiing. She was physiologically very active, and strongly demanded to return to sports. We performed a combined procedure involving a medial closing wedge DFVO using an anatomical locking plate, along with double-bundle ACL reconstruction. The postoperative radiograph confirmed successful correction of knee alignment, specifically achieving varus alignment with precise conformance of the anatomical plate to the medial contour of the distal femur following the osteotomy. The patient resumed her previous sports activities without experiencing knee pain. The operated knee demonstrated restored anterior stability, as indicated by negative Lachman test results, and regained full range of motion. Both the Knee Injury and Osteoarthritis Outcome Score and the 2011 Knee Society score demonstrated continuous postoperative improvements over the three-year follow-up period, indicating positive functional outcomes and joint preservation. To the best of our knowledge, this is the first case of medial closing wedge DFVO with anatomic double-bundle ACL reconstruction in the symptomatic femoral valgus deformity with chronic ACL deficiency in the literature.

A rare case of a testicular lesion related to hand, foot, and mouth disease.

Introduction: Hand, foot, and mouth disease generally occurs in children. In rare cases, hand, foot, and mouth disease affects the testicles. Case presentation: A 29-year-old man presented to our emergency department with testicular pain for several days after the onset of hand, foot, and mouth disease. Ultrasonography revealed hypoechoic mass-like areas in the right testis. A mild inflammatory response was noted, tumor markers and urinary data were normal, and tests for infection were all negative. Antibiotics were initiated and ultrasonography was performed in every subsequent examination. Testicular pain disappeared 6 months later. Conclusion: We encountered a rare case of a testicular lesion related to hand, foot, and mouth disease that was successfully treated. The careful selection of treatment for testicular pain and scrotal enlargement in young adult males, such as surgery and symptomatic treatment, based on their medical history and laboratory findings, is important.

High-risk Features for Recurrence in Patients With Stage III Colorectal Cancer: A Retrospective Cohort Study.

BACKGROUND/AIM: Risk classification for recurrence in stage III colorectal cancer (CRC) is not as well established as it is for stage II. This study aimed to identify high-risk factors for stage III colorectal cancer and to investigate their clinical significance. PATIENTS AND METHODS: We retrospectively analyzed data from 120 patients with stage III CRC who had undergone curative colectomy at our institution between 2014 and 2020. We used logistic regression analysis to identify risk factors for recurrence and subsequently explored their clinical significance. RESULTS: We identified three high-risk factors in stage III CRC: preoperative bowel obstruction [odds ratio (OR)=5.39; 95% confidence interval (CI)=1.61-18.03; p=0.007], N2 disease (OR=3.12; 95%CI=1.05-9.29; p=0.041), and having fewer than 17 examined lymph nodes (OR=3.17; 95%CI=1.11-8.99; p=0.031). The prognosis of patients was clearly stratified by the number of these risk factors, and furthermore, the effectiveness of adjuvant therapy depended on their number. CONCLUSION: Tumor obstruction, N-stage, and the number of lymph nodes examined are important high-risk features for recurrence. This study provides clinicians with valuable insights to predict and stratify patient outcomes in stage III CRC.

Simvastatin Enhances the Radiosensitivity of Androgen-independent Prostate Cancer Cells via Inhibition of RAD51 Expression.

BACKGROUND/AIM: Statins exert antitumor effects via various mechanisms. Additionally, the recurrence rate of prostate cancer after radiation therapy is lower in patients taking statins. This study investigated the efficacy of combination therapy with statins and irradiation in androgen-independent prostate cancer cells. MATERIALS AND METHODS: PC-3 and LNCaP human prostate cancer cell lines were used in this study. We developed androgen-independent LNCaP cells (LNCaP-LA) by gradually replacing fetal bovine serum (FBS) with charcoal-stripped FBS. Microarray analysis was performed, followed by Ingenuity Pathway Analysis. Cell viability was determined using the MTS assay. RESULTS: Simvastatin alters gene expressions in PC-3 cells. Microarray data showed that the number of differentially expressed genes was the highest in the pathway of "Role of BRCA1 in DNA Damage Response". In the validation, the expression of RAD51, listed in "Role of BRCA1 in DNA Damage Response", decreased significantly by simvastatin in PC-3 cells. Reduction in RAD51 expression following siRNA transfection increased the cytocidal effects of X-ray therapy in PC-3 and LNCaP-LA cells. The combination of simvastatin and irradiation further inhibited cell proliferation compared with monotherapy with either therapy in PC-3 or LNCaP-LA cells. In addition, compared with X-ray monotherapy, the combination of simvastatin and irradiation further enhanced the expression of γH2AX, which is reported to be one of the accurate markers of DNA damage in PC-3 cells. CONCLUSION: Simvastatin decreased the expression of RAD51 in androgen-independent prostate cancer cells. The combination of irradiation and drugs that reduce RAD51 expression can potentially affect androgen-independent prostate cancer growth.

Evaluation of the Anterior Cruciate Ligament Injury Risk During a Jump-Landing Task Using 3-Dimensional Kinematic Analysis Versus the Landing Error Scoring System.

Background: The Landing Error Scoring System (LESS) has been utilized on the field or in the clinic to identify patients with an increased risk for anterior cruciate ligament (ACL) injuries; however, its validity and efficacy have not been fully confirmed. Purpose: To assess the efficacy of the LESS in identifying the ACL injury risk by examining the correlation between the LESS score and motion patterns on 3-dimensional kinematic analysis. Study Design: Controlled laboratory study. Methods: The jump-landing motion was analyzed for 16 female basketball or badminton players who volunteered to participate in the study. All study participants were aged 19 or 20 years. The sequence of motion was evaluated with the LESS, while kinematic data were simultaneously acquired with a 3-dimensional motion analysis system utilizing the point cluster method. The correlation between the LESS score and knee kinematics was statistically analyzed. Results: When a LESS score ≥6 was defined to be a risk factor for ACL injuries, 7 of the 16 participants (43.8%) were found to exhibit risky motion patterns. Significant correlations were noted between the LESS score and knee valgus (r = 0.87; P < .0001) and internal tibial rotation (r = 0.57; P = .02) at landing. By contrast, a substantial variability was present in knee flexion, showing no correlation with the LESS score. Conclusion: Significant correlations were found between the LESS score and knee valgus and internal tibial rotation during a jump-landing task. Clinical Relevance: The LESS can be regarded as an effective measure to identify risky motion patterns that may increase the likelihood of ACL injuries.

Simvastatin Induces Autophagy and Inhibits Proliferation in Prostate Cancer Cells.

BACKGROUND/AIM: Statin has recently been studied for its effects on inducing cell death and inhibiting metastasis. Nevertheless, the precise mechanism of its anti-tumor effect is not yet fully understood. We conducted research on statin as a novel treatment for castration-resistant prostate cancer (CRPC). This study focused on autophagy in prostate cancer cells and assessed the effects of simvastatin. MATERIALS AND METHODS: After administering simvastatin to PC-3 cells, we conducted a microarray analysis. Simvastatin was administered to prostate cancer cell lines (PC-3, LNCaP-LA; cultured under androgen-depleted conditions, DU145, 22RV1), and the tumor proliferation inhibition was evaluated using the MTS assay and cell count. Autophagy was measured by observing autophagosome staining under a fluorescence microscope and quantifying LC-3 protein using western blot. We also investigated the effects of rapamycin, an autophagy inducer, and chloroquine as an inhibitor. RESULTS: Simvastatin demonstrated a significant concentration-dependent growth inhibition effect on prostate cell lines. Moreover, a significant increase in autophagy was observed in all cell lines following simvastatin administration. When we administered simvastatin with rapamycin at a concentration that did not show a tumor growth inhibitory effect, it significantly enhanced autophagy induction compared to simvastatin alone, and also significantly enhanced the growth inhibition effect on PC-3 cells. CONCLUSION: Simvastatin induced autophagy and inhibited the proliferation of prostate cancer cell lines. The combination of simvastatin and rapamycin significantly induced autophagy and enhanced the inhibitory effect of simvastatin on proliferation. This mechanism may serve as a novel therapeutic target.

Osteochondral autologous transplantation for large steroid-induced osteonecrosis lesion of the knee in a young patient yielding satisfactory results despite only partial coverage: a case report.

Osteochondral autologous transplantation (OAT) is one of the most common surgical options for osteochondral disorders of the knee. In cases where OAT is performed for steroid-induced osteonecrosis, there are several problems potentially affecting the surgical outcomes such as large chondral damage area and compromised host bone. In addition, steroid administration for a long period of time may lead to extensive lesion, which poses difficulty in obtaining sufficient donor tissue. Those factors affect the prognosis of steroid-induced osteonecrosis resulting in inferior treatment outcomes. We present a young female with a large steroid-induced osteonecrosis lesion repaired only with two osteochondral plugs harvested from the healthy area. The reported case indicates that only partial osteochondral grafting limiting to the weight-bearing area may yield satisfactory outcome when OAT is performed for large steroid-induced osteonecrosis of the knee.

High prevalence of associated intra-articular injuries with increasing time period from anterior cruciate ligament injury to reconstruction, even in apparent copers.

BACKGROUND: It still unclear whether copers may bear the same time-sensitive risk of intra-articular injury as non-copers. The objectives were to investigate the clinical characteristics of copers and non-copers that have sustained an anterior cruciate ligament (ACL) injury, and to examine and compare the intra-articular pathologies in delayed ACL reconstruction (ACLR) in copers and noncopers. METHODS: Patients who sustained ACL injury while participating in high-performance sports and opted for non-operative treatment were enrolled in this study. Depending on the occurrence of the knee giving way, patients were classified into copers and noncopers. Clinical characteristics were compared between the two groups. Additionally, intra-articular injuries were evaluated for those who eventually underwent delayed ACLR. RESULTS: 11 of the 75 patients (14.7%) were classified as copers. No major differences were found in the clinical characteristics between groups. Following the initial non-operative treatment, 67 patients underwent delayed ACLR. When examining intra-articular abrasions at the time of surgery, non-copers who continued sports activities for 3 to 12 months exhibited a significantly higher rate of injury as opposed to their coper counterparts. However, the difference in the prevalence of intra-articular lesions between the two groups in patients who continued to play sports for at least 12 months before surgery was nominal. CONCLUSION: The rate of copers was relatively low in patients who resumed playing high-level sports after ACL injury. Additionally, even in copers, those who continued sports activities for more than 12 months had comparably high prevalence of intra-articular injuries with noncopers. STUDY DESIGN: Retrospective case-control study; Level of evidence, 3.

Optimal additional support screw position for prevention of hinge fracture in biplanar closed wedge distal femoral osteotomy.

Background: The purpose of this study was to examine the biomechanical significance of supplemental fixation using a positional screw in prevention of the hinge fracture in lateral closed-wedge distal femoral osteotomy (LCW-DFO) by means of a three-dimensional finite element analysis. Methods: The three-dimensional numerical knee models with LCW-DFO were developed. To assess the mechanical efficacy of the positional screw and determine its optimal position and orientation, in total, 13 screwing methods were analyzed. In the first four methods, the screw was supported by the cortical bone only on the medial surface (mono-cortical). In the other 9 models, the screw was supported by both medial and lateral cortical bones (bi-cortical). Under 1000 N of vertical force and 5 Nm of rotational torques, the highest shear stress value around the medial hinge area was adopted as an analytical parameter. Results: In mono-cortical methods, with the cancellous bone support, all methods were able to reduce the highest stress value compared to the value without the screw, while the efficacy was rather inferior when the screw was in horizontal direction. Without the cancellous bone support, however, all methods were not able to reduce the stress value. In bi-cortical methods, with the cancellous bone support, almost all screw augmentation methods were able to reduce the stress value. When screwing from the medial to the lateral, it only gets worse when going extremely posterior. Without the cancellous bone support, all methods were able to reduce the stress value. Conclusion: The mechanical efficacy of the bi-cortical method was proven regardless of the quality of the local cancellous bone.

Postoperative Tunnel Widening, Elliptical Aperture Shape, and No Preservation of the Remnant Are Related to the Tendon Graft-Bone Tunnel Gap Formation at the Intra-Articular Aperture After Double-Bundle Anterior Cruciate Ligament Reconstruction.

Purpose: To examine the bone-tendon healing at the posterolateral (PL) femoral tunnel aperture by second-look arthroscopy after double-bundle anterior cruciate ligament reconstruction (ACLR), and assess the risk factors for impaired healing at the tendon-bone interface. Methods: A consecutive series of knees undergoing primary double-bundle ACLR using hamstring tendon autografts were enrolled in the study. The exclusion criteria were as follows: previous knee surgeries, concomitant ligamentous and osseous procedures, and a lack of second-look arthroscopy or postoperative computed tomography data for the analysis. Cases in which a gap was identified between the graft and tunnel aperture during the second-look arthroscopic examination were classified as the gap formation (GF) group. A multivariate logistic regression analysis was performed to assess the relationship between the GF and variables that may determine prognosis. Results: A total of 54 knees that met the inclusion/exclusion criteria were included in the study. Second-look arthroscopy revealed the GF at the PL aperture in 22 of the 54 knees (40%). The time period from surgery to arthroscopy averaged 16 months. In the multivariate logistic regression analysis, the percentage tunnel widening at 1 year on computed tomography (odds ratio, 10.4; 95% confidence interval [CI] 1.56-69.2), ellipticity of the tunnel aperture (odds ratio, 3.57; 95% CI, 0.79-16.11), and no ACL remnant preservation (odds ratio, 5.99; 95% CI, 1.23-29.06) were identified as prognostic factors significantly related to graft-bone tunnel GF. Conclusions: Second-look arthroscopy revealed GF at the PL graft-bone tunnel interface in 40% of the knees after double-bundle ACLR. Incomplete healing of the interface, as evidenced by a graft-bone gap at the tunnel aperture, was associated with tunnel widening 1-year postsurgery, an elliptical aperture shape, and no preservation of the ACL remnant. Level of Evidence: Ⅲ, retrospective case-control study.

Cryo-EM structure of the transposon-associated TnpB enzyme.

The class 2 type V CRISPR effector Cas12 is thought to have evolved from the IS200/IS605 superfamily of transposon-associated TnpB proteins1. Recent studies have identified TnpB proteins as miniature RNA-guided DNA endonucleases2,3. TnpB associates with a single, long RNA (ωRNA) and cleaves double-stranded DNA targets complementary to the ωRNA guide. However, the RNA-guided DNA cleavage mechanism of TnpB and its evolutionary relationship with Cas12 enzymes remain unknown. Here we report the cryo-electron microscopy (cryo-EM) structure of Deinococcus radiodurans ISDra2 TnpB in complex with its cognate ωRNA and target DNA. In the structure, the ωRNA adopts an unexpected architecture and forms a pseudoknot, which is conserved among all guide RNAs of Cas12 enzymes. Furthermore, the structure, along with our functional analysis, reveals how the compact TnpB recognizes the ωRNA and cleaves target DNA complementary to the guide. A structural comparison of TnpB with Cas12 enzymes suggests that CRISPR-Cas12 effectors acquired an ability to recognize the protospacer-adjacent motif-distal end of the guide RNA-target DNA heteroduplex, by either asymmetric dimer formation or diverse REC2 insertions, enabling engagement in CRISPR-Cas adaptive immunity. Collectively, our findings provide mechanistic insights into TnpB function and advance our understanding of the evolution from transposon-encoded TnpB proteins to CRISPR-Cas12 effectors.

Efficacy of intravenous tranexamic acid administration in medial opening-wedge distal tibial tuberosity osteotomy (MOWDTO) for varus knee osteoarthritis: a randomized control trial.

BACKGROUND: This randomized controlled study was undertaken to investigate the efficacy of intravenous tranexamic acid (TXA) administration in reducing perioperative blood loss in patients undergoing medial opening-wedge distal tibial tuberosity osteotomy (MOWDTO). It was hypothesized that TXA would reduce perioperative blood loss in MOWDTO. METHODS: A total of 61 knees in 59 patients who underwent MOWDTO during the study period were randomly assigned to either of the groups with intravenous TXA administration (TXA group) or without TXA administration (control group). In the TXA group, patients received 1000 mg of TXA intravenously before skin incision and 6 h after the first dose. The primary outcomes was the volume of perioperative total blood loss which calculated using the blood volume and hemoglobin (Hb) drop. The Hb drop was calculated as the difference between preoperative Hb and postoperative Hb at days 1, 3, and 7. RESULTS: The perioperative total blood loss was significantly lower in the TXA group (543 ± 219 ml vs. 880 ± 268 ml, P < 0.001). The Hb drop was significantly lower at postoperative days 1, 3 and 7 in the TXA group than in the control group (day 1: 1.28 ± 0.68 g/dl vs. 1.91 ± 0.69 g/dl, P = 0.001; day 3: 1.54 ± 0.66 g/dl vs. 2.69 ± 1.00 g/dl, P < 0.001; day 7: 1.74 ± 0.66 g/dl vs. 2.83 ± 0.91 g/dl, P < 0.001). CONCLUSION: Intravenous TXA administration in MOWDTO could reduce the perioperative blood loss. Trial registration The study was approved by the institutional review board. (Registered on 26/02/2019 Registration Number 3136). Level of Evidence Level I, randomized controlled trial.

Effect of Cross-sectional Area of the Hamstring Tendon Autograft on Failure Rate or Clinical Outcomes After Double-Bundle ACL Reconstruction: Tendon Autograft.

Background: In previous studies examining the relationship between graft size and failure rate after anterior cruciate ligament reconstruction (ACLR), graft size was determined as diameter of the bone tunnel, and graft failure was defined as revision surgery. Consequently, the correlation between graft size and postoperative recurrent instability could not be assessed. Purpose: (1) To intraoperatively measure the cross-sectional area (CSA) of the hamstring tendon (HT) autograft and compare the CSA of the autograft with the bone tunnel and (2) to assess the effect of the graft CSA on postoperative graft failure among patients who underwent double-bundle ACLR. Study Design: Case-control study; Level of evidence, 3. Methods: The study included 129 patients who underwent double-bundle ACLR using an HT autograft (mean ± SD age, 16.7 ± 1.7 years; all with a Tegner activity level ≥6). All patients had a minimum follow-up of 2 years. During surgery, the graft CSA was measured using an area micrometer, combining the anteromedial (AM) and posterolateral (PL) grafts. The total area of the bone tunnel was defined as the combined CSAs of the AM and PL tunnels as calculated by the diameter of the drill. The relationship between the CSAs of the combined HT graft and the bone tunnel was statistically compared, as was the relationship between graft CSAs and graft failure, defined as reinjury, recurrent instability manifested as quantitative laxity measurement, or revision ACLR. Results: The CSAs of the midsubstance of the combined AM and PL graft significantly correlated with those of the bone tunnels (femoral side, R 2 = 0.334, P < .0001; tibial side, R 2 = 0.421, P < .0001). As for the relationship between the graft CSA and ACLR failure, there was no significant difference in the graft CSAs between the groups with and without graft failure in any of the failure criteria (P = .188). Conclusion: The graft CSA was not a predictor of early failure after double-bundle ACLR using an HT autograft in this patient population.

Preoperative Ahlbäck radiographic classification grade significantly influences clinical outcomes of double level osteotomy for osteoarthritic knees with severe varus deformity.

PURPOSE: The purpose of this study was to examine the relationship between preoperative Ahlbäck radiographic classification grade and the clinical outcomes of double level osteotomy (DLO) performed for osteoarthritic knees with severe varus deformity. METHODS: The study population comprised a consecutive series of 99 knees (68 patients) for which DLO was performed and follow-up results for a minimum of two years were available. The Ahlbäck radiographic classification system was used to determine the osteoarthritic grade. The following radiological parameters for alignment and bone geometry were measured: mechanical lateral distal femoral angle (mLDFA), mechanical medial proximal tibial angle (mMPTA), joint-line convergence angle (JLCA), and mechanical tibiofemoral angle (mTFA). Clinical results were assessed using the Knee Injury and Osteoarthritis Outcome Score (KOOS) and the International Knee Documentation Committee (IKDC) subjective score preoperatively and at 2 years after surgery. Difference between preoperative and postoperative measurements as well as relationship between Ahlbäck grade and radiological/clinical results were statistically assessed. RESULTS: The average age of the study participants was 60.9 ± 6.2 years and the mean follow-up period was 45.4 ± 15.2 months. Each of the radiological parameters exhibited preoperative abnormal values. Knees with Ahlbäck grade 3 and 4 osteoarthritis exhibited significantly greater JLCA and mTFA than grade 1 knees. Two years post-surgery, all radiological parameter values measured within a normal range. Clinical evaluation showed significant improvement in KOOS after surgery. Analysis of the relationship between Ahlbäck grade and clinical score showed that the 2-year postoperative KOOS scores in grade 3 and 4 osteoarthritic knees were significantly lower than grade 1 knees (with the mean 2-year KOOS scores of 350.0 ± 79.9, 317.9 ± 78.3, and 420.2 ± 42.9, respectively). CONCLUSIONS: While DLO may produce significant radiological and clinical improvement in knees with joint space obliteration, Ahlbäck grade 3 and 4 osteoarthritic knees associated with larger JLCA and mTFA showed less satisfactory clinical results compared to grade 1 knees. LEVEL OF EVIDENCE: IV case series.

Liquid Chromatography-Mass Spectrometry-Based Qualitative Profiling of mRNA Therapeutic Reagents Using Stable Isotope-Labeled Standards Followed by the Automatic Quantitation Software Ariadne.

mRNA-based medicines are a promising modality for preventing virus-caused illnesses, including COVID-19, and treating various types of cancer and genetic diseases. To develop such medicines, methods to characterize long mRNA molecules are needed for quality control and metabolic analysis. Here, we developed an analytical platform based on isotope-dilution liquid chromatography-mass spectrometry (LC-MS) that quantitatively characterizes long, modified mRNAs by comparing them to a stable isotope-labeled reference with an identical sequence to that of the target medicine. This platform also includes database searching using the mass spectra as a query, which allowed us to confirm the primary structures of 200 to 4300 nt mRNAs including chemical modifications, with sequence coverage at 100%, to detect/identify defects in the sequences, and to define the efficiencies of the 5'-capping and integrity of the polyadenylated tail. Our findings indicated that this platform should be valuable for quantitatively characterizing mRNA vaccines and other mRNA medicines.

Increased posterior slope and coronal inclination of the tibial joint line after opening wedge high tibial osteotomy may induce mucoid degeneration of the anterior cruciate ligament: A case report.

A mucoid degeneration of the anterior cruciate ligament (ACL) is regarded as a degenerative change in the ligament, which is clinically presented with pain on full extension or flexion. Regarding morphological factors, it has been reported that an increased posterior tibial slope can be a cause of ACL degeneration secondary to the repetitive overload. The increase in the tibial slope is among the potential problems after medial opening wedge high tibial osteotomy (OWHTO). Especially, a large wedge opening in the correction of severe varus deformity may lead to non-physiologic bony geometry including an increased posterior tibial slope and medial tibial coronal inclination. We present a 69-year-old man had undergone OWHTO with a wedge correction angle of 12.4° for Kellegren-Lawrence grade 2, medial uni-compartmental osteoarthritis of the left knee. Evaluations of the postoperative radiographs revealed postoperative changes in radiological parameters with mechanical medial proximal tibial axis (mMPTA) from 81.3° to 94.3°, and posterior tibial slope (PTS) from 12.2° to 15.8°. Physical examination at 3 years after surgery revealed a knee extension of 0° and a limitation to knee flexion with maximum flexion of 110° and, and severe knee pain was elicited when the knee approached deep flexion. MRI revealed an increased signal intensity along the substance of the ACL and multiple cystic lesions indicative of a ganglion formation around the proximal ACL attachment site extending into the adjacent lateral femoral condyle. Microscopic examination of the resected tissues showed mucoid degeneration and mucous cysts indicative of ganglions formation within the ligament substance and the bone at the attachment site. The reported case illustrates the importance of being aware of this potential complication following OWHTO.

An alpaca single-domain antibody (VHH) phage display library constructed by CDR shuffling provided high-affinity VHHs against desired protein antigens.

Antigen-combining sites of the camelid heavy-chain antibody variable domain (VHH) are constructed by three complementarity-determining regions (CDR1, CDR2 and CDR3). We prepared cDNA using mRNA extracted from peripheral lymphocytes of alpacas that had been non-immunized or immunized with human serum albumin (HSA). The VHH gene fragments encoding the amino-terminal half-containing CDR1 as well as CDR2 and the carboxy-terminal half-containing CDR3 were amplified independently by PCR, and then full-length VHH gene fragments were generated by overlap extension PCR and cloned into the phagemid vector. This protocol, referred to as CDR shuffling, allowed us to construct an alpaca VHH phage display library possessing repertoires different from those naturally occurring in animals. We asked, first, whether this library was able to provide the functional VHH fragments against HSA, an immunized antigen, and obtained 29 anti-HSA VHH clones, 41% possessed KD values of lower than 10-8 M, 5 of which had KD values of 10-10 M. We also obtained VHH clones against non-immunized protein antigens such as cardiac troponin T and I, Ebola virus glycoprotein 1 and human immunoglobulin G by biopanning. We compared the amino acid sequences and affinities and found that 43% of VHHs had KD values of less than 10-8 M, although those having KD values of 10-10 M were unavailable. These results suggested that the CDR-shuffled VHH phage display library could potentially provide VHHs against non-immunized protein antigens with similar levels of affinities to those against immunized antigens.

Tibial Condylar Valgus Osteotomy Combined With Medial Open-Wedge Distal Tuberosity Tibial Osteotomy.

Severe varus deformity in osteoarthritic knees is attributed not only to bony deformity but also to intra-articular deformity as a result of medial joint space narrowing with lateral joint space widening (increased joint line convergence angle). In such knees, correction of bony deformity by high tibial osteotomy (HTO) alone may not be capable of restoring physiological joint geometry and biomechanics. Tibial condylar valgus osteotomy (TCVO), an L-shaped osteotomy in the medial tibial condyle, has been proposed to improve articular stability and congruity by elevating the medial tibial joint line and addressing the articular component of the deformity; however, its surgical efficacy for correction of the valgus deformity is limited. Therefore a procedure combining HTO and TCVO can be a reasonable option to achieve restoration of the knee joint physiology in such situations. In our current practice, medial open-wedge distal tuberosity tibial osteotomy (MOW-DTO) has been the procedure of primary option among the HTO procedures to avoid patellofemoral overload which could be an inherent problem in medial open-wedge HTO. In this article, the surgical rationale and the combined procedure of TCVO and MOW-DTO are described.

Combination therapy with novel androgen receptor antagonists and statin for castration-resistant prostate cancer.

BACKGROUND: One of the growth mechanisms of castration-resistant prostate cancer (CRPC) is de novo androgen synthesis from intracellular cholesterol, and statins may be able to inhibit this mechanism. In addition, statins have been reported to suppress the expression of androgen receptors (ARs) in prostate cancer cell lines. In this study, we investigated a combination therapy of novel AR antagonists and statin, simvastatin, for CRPC. METHODS: LNCaP, 22Rv1, and PC-3 human prostate cancer cell lines were used. We developed androgen-independent LNCaP cells (LNCaP-LA). Microarray analysis was performed, followed by pathway analysis, and mRNA and protein expression was evaluated by quantitative real-time polymerase chain reaction and Western blot analysis, respectively. Cell viability was determined by MTS assay and cell counts. All evaluations were performed on cells treated with simvastatin and with or without AR antagonists (enzalutamide, apalutamide, and darolutamide). RESULTS: The combination of darolutamide and simvastatin most significantly suppressed proliferation in LNCaP-LA and 22Rv1 cells. In a 22Rv1-derived mouse xenograft model, the combination of darolutamide and simvastatin enhanced the inhibition of cell proliferation. In LNCaP-LA cells, the combination of darolutamide and simvastatin led to reduction in the mRNA expression of the androgen-stimulated genes, KLK2 and PSA; however, this reduction in expression did not occur in 22Rv1 cells. The microarray data and pathway analyses showed that the number of differentially expressed genes in the darolutamide and simvastatin-treated 22Rv1 cells was the highest in the pathway termed "role of cell cycle." Consequently, we focused our efforts on the cell cycle regulator polo-like kinase 1 (PLK1), cyclin-dependent kinase 2 (CDK2), and cell cycle division 25C (CDC25C). In 22Rv1 cells, the combination of darolutamide and simvastatin suppressed the mRNA and protein expression of these three genes. In addition, in PC-3 cells (which lack AR expression), the combination of simvastatin and darolutamide enhanced the suppression of cell proliferation and expression of these genes. CONCLUSIONS: Simvastatin alters the expression of many genes involved in the cell cycle in CRPC cells. Thus, the combination of novel AR antagonists (darolutamide) and simvastatin can potentially affect CRPC growth through both androgen-dependent and androgen-independent mechanisms.