Fracture-Dedicated Prosthesis Promotes the Healing Rate of Greater Tuberosity in Reverse Shoulder Arthroplasty: A Meta-Analysis.

Introduction: Reverse shoulder arthroplasty (RSA) is becoming popular in the treatment of complex proximal humeral fractures (PHFs). Greater tuberosity healing may influence functional outcomes and range of motion (ROM) of shoulder after RSA. In addition, the design of prosthesis may impact the healing rate of greater tuberosity. The purpose of this study is to know: (1) does the healing of greater tuberosity affect the functional outcomes and ROM of shoulder? and (2) does the design of prosthesis affect the healing rate of greater tuberosity? Materials and Methods: PubMed, Ovid/Embase, and the Cochrane Library were searched for studies comparing the clinical outcomes between the healed groups and the non-healed groups after RSA. Results: For functional outcomes, the results showed that the healed group had better Constant scores (CSs) (p < 0.0001). For ROM, the healed group showed better flexion (p < 0.0001), abduction (p = 0.02), and external rotation (p < 0.00001) of shoulder. For the design of prosthesis, the mean healing rate of greater tuberosity (82.7%) in patients with fracture-dedicated prosthesis was higher than those (63.0%) in patients with standard prosthesis. Subgroup analyses showed that the CS (p = 0.12) and abduction (p = 0.96) of patients using fracture-dedicated prostheses were not different between the healed groups and the non-healed groups. Meta-regression showed that there was no significant relationship between the design of prosthesis and CS (p = 0.312), flexion (p = 0.422), or external rotation (p = 0.776). Conclusion: Our meta-analysis showed that the healed groups could obtain better functional outcomes and ROM than the non-healed groups. In addition, fracture-dedicated prostheses promoted the healing rate of greater tuberosity. Systematic Review Registration: https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42020157276, PROSPERO: CRD42020157276.

A Stable Large Animal Model for Dural Defect Repair with Biomaterials and Regenerative Medicine.

IMPACT STATEMENT: Using biomaterials and regenerative medicine to repair tissue defects has been a very hot research field, during which the development of stable large animal models with appropriate biotechnology is crucial. Recently, more and more researchers are paying attention to dural defect repair. However, the lack of widely recognized stable large animal models has seriously affected the related further research. In this study, a stable large animal dural defect model is developed exactly for the first time. Therefore, the article would attract considerable attention and be highly cited after publication.

COPI-Mediated Nuclear Translocation of EGFRvIII Promotes STAT3 Phosphorylation and PKM2 Nuclear Localization.

As a non-ligand-dependent activation protein, EGFRvIII is the most common mutant of EGFR, and its existence or especially its nuclear translocation in tumors can exacerbate the malignancy. Compared with the nuclear translocation of EGFR, which has been studied extensively, the specific mechanism by which EGFRvIII undergoes nuclear translocation has not yet been reported. Here, we found that EGFRvIII eventually reached the nucleus with the involvement of the Golgi and endoplasmic reticulum (ER) in glioma cells. In this process, syntaxin-6 was responsible for the identification and transport of EGFRvIII on Golgi. We also demonstrated that COPI mediated the reverse transport of EGFRvIII from the Golgi to ER, which process was also important for EGFRvIII's nuclear accumulation. EGFRvIII's nuclear translocation can significantly promote STAT3 phosphorylation and PKM2 nuclear localization. Finally, we showed that EGFRvIII's nuclear translocation obviously induced the growth of gliomas in an intracranial xenotransplantation experiment. These data suggested that searching methods that inhibit EGFRvIII entry into the nucleus will be effective glioma treatments.

Combined use of Kirschner wires and hinged external fixator for capitellar and trochlear fractures: a minimum 24-month follow-up.

BACKGROUND: Open reduction and internal fixation is the adequate treatment for capitellar and trochlear fractures. Given the low incidence of this type of fractures, it is difficult to constitute a universally accepted method for fixation. Thus, we hypothesised that combined use of Kirschner wires (K-wires), absorbable rods and sutures for fixation and post-operative hinged external fixator for early rehabilitation exercise can restore elbow joint function well. METHODS: This retrospective study included 20 patients with a mean age of 48.3 (range 16-76) years. According to the Dubberley classification, fractures were classified on plain radiographs, computed tomography images and intra-operative findings. All patients were evaluated by the range of motion of the elbow and the Broberg-Morrey score. RESULTS: All fractures had healed without non-union, and the average time was 13.6 (range 8-17) weeks. The mean follow-up was 42.5 (range 24-80) months. The mean flexion was 117.1° (range 90°-135°), and the mean extension was 17.5° (range 0°-45°). The mean pronation was 74.4° (range 45°-85°), and the mean supination was 84.3° (range 60°-90°). The average Broberg-Morrey score was 86.2 (range 68-98) points with 10 excellent, 7 good and 3 fair results. CONCLUSION: K-wires, absorbable rods and sutures combined with hinged external fixator are feasible for fixation of capitellar and trochlear fractures. However, due to the absence of a control group (such as Herbert screw fixation), comparative studies are still needed to demonstrate the safety and reliability of K-wires for fixation.

Enhanced intrinsic CYP3A4 activity in human hepatic C3A cells with optically controlled CRISPR/dCas9 activator complex.

Human hepatic C3A cells have been applied in bioartificial liver development, although these cells display low intrinsic cytochrome P450 3A4 (CYP3A4) enzyme activity. We aimed to enhance CYP3A4 enzyme activity of C3A cells utilizing CRISPR gene editing technology. We designed two CYP3A4 expression enhanced systems applying clustered regularly interspaced short palindromic repeats (CRISPR) gene technology: a CRISPR-on activation system including dCas9-VP64-GFP and two U6-sgRNA-mCherry elements, and a light-controlled CRISPR-on activation system combining our CRISPR-on activation system with an optical control system to facilitate regulation of CYP3A4 expression for various applications. Results of enzymatic activity assays displayed increased CYP3A4 activity in C3A cells expressing the CRISPR-on activation system compared with C3A cells. In addition, CYP3A4 activity increased in C3A cells expressing the light-controlled CRISPR-on activation system under blue light radiation compared with C3A cells. Notably, there was no statistical difference in the increase of CYP3A4 protein amounts induced by these two methods. After expansion in culture, C3A cells with the light-controlled CRISPR-on activation system exhibited no statistical difference in CYP3A4 mRNA levels between generations. Our findings provide a method to stably enhance functional gene expression in bioartificial liver cells with the potential for large-scale cell expansion.

The impact of associated injuries and fracture classifications on the treatment of capitellum and trochlea fractures: A systematic review and meta-analysis.

BACKGROUND: Capitellum and trochlea fractures are truly rare and the treatment is not fully appreciated. So we evaluate the impact of associated injuries and fracture classifications on elbow functional outcomes after open reduction and internal fixation. MATERIALS AND METHODS: PubMed, Embase, Ovid Medline, and the Cochrane Library were searched from January 1, 1974 to January 1, 2017. All English literature with the treatment of capitellum and trochlea fractures by open reduction and internal fixation were included. RESULTS: For associated injuries, the results suggested that the MEPI score of patients without associated injuries was higher than that of patients with associated injuries (P = 0.001). However, there was no significant difference in the arc of motion between the two groups (P = 0.052). For Bryan and Morrey classification, there was no significant difference in the MEPI score (P = 0.622) and in the arc of motion (P = 0.652) between type-I fractures and type-IV fractures. For Dubberley classification, there was significant difference only in the MEPI score between subtype-A fractures and subtype-B fractures (P = 0.005). CONCLUSION: The associated injury of fracture may have a negative impact on the functional outcomes of elbow. And Dubberley classification is more suitable to classify this kind of fracture. Furthermore, high-quality studies are required to attain robust evidence.

Comparison of small intestinal submucosa and polypropylene mesh for abdominal wall defect repair.

Abdominal wall defects are a common medical problem, and inadequate repair methods can lead to serious complications. Abdominal wall reconstruction using autologous tissue, or non-biological, biological, or composite patches is often performed to repair defective areas. In particular, composite patches containing both polymeric and biological materials have gained increasing attention due to their good mechanical properties and biocompatibility. However, it is still unclear whether the quality of repairs using composite patches is superior to that of a biological patch. Based on the limitations of previous studies, we compared small intestinal submucosa (SIS) patches with SIS + polypropylene mesh (PPM) patches for repairing abdominal wall defects in adult beagle dogs. Forty-five female dogs were subjected to surgical resection to produce abdominal wall defects. SIS or SIS + PPM was used as patch for the defects. Morphology, biomechanics, and histological evaluations were performed to evaluate the efficacy and safety of such therapies. Our findings demonstrated that SIS had advantages over SIS + PPM considering biological activity and histocompatibility without increasing the risk of repair failure.

Efficacy and safety of small intestinal submucosa in dural defect repair in a canine model.

Dural defects are a common problem, and inadequate dural closure can lead to complications. Several types of dural substitute materials have recently been discarded or modified owing to poor biocompatibility or mechanical properties and adverse reactions. The small intestinal submucosa (SIS) is a promising material used in a variety of applications. Based on the limitations of previous studies, we conducted an animal study to evaluate the efficacy and safety of the SIS in preclinical trials. Twenty-four male beagle dogs were subjected to surgical resection to produce dural defects. SIS or autologous dural mater was patched on the dural defect. Gross and histological evaluations were carried out to evaluate the efficacy and safety of the therapy. Our findings demonstrated that the SIS, which stimulated connective and epithelial tissue responses for dural regeneration and functional recovery without immunological rejection, could provide prolonged defect repair and prevent complications. The mechanical properties of the SIS could be adjusted by application of multiple layers, and the biocompatibility of the material was appropriate. Thus, our data suggested that this material may represent an alternative option for clinical treatment of dural defects.

Resistance of glioma cells to nutrient-deprived microenvironment can be enhanced by CD133-mediated autophagy.

CD133 is a pentaspan transmembrane protein that can serve as a biomarker for cancer stem cells, although its biochemical mechanism remains unclear. Here we report that CD133 expression enhances glioma cell tolerance of a nutrient-deprived microenvironment. Under starvation conditions, CD133-positive cells exhibited higher survival and decreased levels of apoptosis. These changes were dependent on activation of autophagy-associated gene signaling and were impaired by the autophagic inhibitor chloroquine. Furthermore, rapamycin up-regulated the level of autophagy and inversely reduced CD133 expression. Immunofluorescence confirmed that starvation promoted release of CD133 from the plasma membrane to the cytoplasm, with CD133 also partially co-localizing with LC3 upon starvation. Additionally, CD133 partially co-localized with Beclin1, Atg5, and lysosomes, indicating that CD133 directly participates in the autophagosome membrane fusion process and ultimately undergoes lysosomal degradation. Collectively, our results demonstrate that CD133 contributes to cell survival by regulating autophagy, and that targeting CD133-linked signaling and autophagy may be useful in improving anti-cancer treatments.

Knock out CD44 in reprogrammed liver cancer cell C3A increases CSCs stemness and promotes differentiation.

CD44 is a widely known cancer stem cells marker in various cancers and validated to function in tumor growth, survival and tumor metastasis. In this study, we first established C3A-derived liver cancer stem cells by OSKM method [OCT4, SOX2, KLF4, and c-MYC], termed C3A-induced cancer stem cells (C3A-iCSCs) which acquired self-renewal and stemness abilities. Then we found CD44 was positive in C3A-iCSCs and mainly located in cell nuclear. Chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR) results showed nuclear CD44 combined promoter regions of c-MYC and SOX2. These results suggested that CD44 participated in C3A-iCSCs transcriptional regulation. To explore CD44 overall influence in liver cancer stem cells, CD44 was knocked out in C3A-iCSCs using CRISPR/Cas9 technology. Our results showed a dramatic increase in the expression of stem cell markers OCT4, SOX2 and NANOG in CD44- C3A-iCSCs compared with that in CD44+ C3A-iCSCs. Tumor derived from CD44- C3A-iCSCs also displayed well-differentiated tumor cells compared to CD44+ C3A-iCSCs, which suggested CD44- C3A-iCSCs derived tumor cells exhibited lower malignant degree. Our data indicated nuclear CD44 in liver cancer stem cells is responsible for the poorly differentiated highly malignant tumor cells by maintenance of low stemness state.

[RESEARCH PROGRESS OF BIOMECHANICS OF PROXIMAL ROW CARPAL INSTABILITY].

OBJECTIVE: To review the research progress of the biomechanics of proximal row carpal instability (IPRC). METHODS: The related literature concerning IPRC was extensively reviewed. The biomechanical mechanism of the surrounding soft tissue in maintaining the stability of the proximal row carpal (PRC) was analyzed, and the methods to repair or reconstruct the stability and function of the PRC were summarized from two aspects including basic biomechanics and clinical biomechanics. RESULTS: The muscles and ligaments of the PRC are critical to its stability. Most scholars have reached a consensus about biomechanical mechanism of the PRC, but there are still controversial conclusions on the biomechanics mechanism of the surrounding soft tissue to stability of distal radioulnar joint when the triangular fibrocartilage complex are damaged and the biomechanics mechanism of the scapholunate ligament. At present, there is no unified standard about the methods to repair or reconstruct the stability and function of the PRC. So, it is difficult for clinical practice. CONCLUSION: Some strides have been made in the basic biomechanical study on muscle and ligament and clinical biomechanical study on the methods to repair or reconstruct the stability and function of PRC, but it will be needed to further study the morphology of carpal articular surface and the adjacent articular surface, the pressure of distal carpals to proximal carpal and so on.

[EFFECTIVENESS OF LIMITED INTERNAL FIXATION COMBINED WITH HINGED SUPER-ARTICULAR EXTERNAL FIXATOR FOR TYPE C3 FRACTURE OF DISTAL HUMERUS IN ADULT].

OBJECTIVE: To investigate the effectiveness of limited internal fixation combined with hinged super-articular external fixator to treat type C3 fracture of the distal humerus. METHODS: Between September 2007 and November 2012, 37 cases of type C3 fracture of the distal humerus were treated. There were 22 males and 15 females with an average age of 43.6 years (range, 22-66 years). The causes were accident injury in 24 cases, falling injury in 5 cases, falling from height in 4 cases, heavy crush injury in 2 cases, machine injury in 1 case, and other injury in 1 case. There were 22 cases of open injury and 15 cases of closed injury. The time from injury to operation was 3-46 hours (mean, 18 hours). RESULTS: Needle tract reaction and incision infection occurred in 3 cases and 1 case respectively, healing of incision by first intension was obtained in the other cases. Thirty-six patients were followed up 9-48 months (mean, 25.4 months). Heterotopic ossification occurred in 3 cases after operation and no recurrence was found by release after fracture healing. Fractures healed in the other patients after 6-14 months (mean, 9 months) of operation except 1 patient who suffered chronic osteomyelitis. One patient had delayed ulnar neuritis at 12 months after operation, and the nerve function returned to normal after the ulnar nerve transposition. There was no bone ischemic necrosis, elbow joint instability, or loosening of internal fixation. At last follow-up, the average range of motion of injured elbow was 105.0° in flexion, -25.0° in extension, 69.2° in pronation, and 75.6° in supination. According to Mayo elbow joint function score (MEPS) and disability of arm shoulder and hand (DASH) score, the results were excellent in 22 cases, good in 8 cases, fair in 4 cases, and poor in 2 cases with an excellent and good rate of 83.3%; and according to Cassebaum elbow joint function score, the results were excellent in 21 cases, good in 7 cases, fair in 5 cases, and poor in 3 cases with an excellent and good rate of 77.8%. CONCLUSION: A combination of limited internal fixation and hinged super-articular external fixator has satisfactory clinical curative effect for type C3 fractures of the distal humerus, relatively few complications.

Adenovirus vector-mediated expression of TMEM166 inhibits human cancer cell growth by autophagy and apoptosis in vitro and in vivo.

TMEM166 is a novel programmed cell death-related molecule. In this report, we constructed a recombinant adenovirus 5-TMEM166 vector (Ad5-TMEM166) and evaluated its expression and anti-tumor activities in vitro and in vivo. Cell viability analysis revealed that the adenovirus-mediated increase of TMEM166 inhibited tumor cell growth in a dose- and time-dependent manner. This inhibitory effect was mediated by both autophagy (via inhibition of mTOR and activation of p70S6K) and apoptosis (via caspase-3 activation), both of which contributed to cell death and suppression of tumorigenicity. Our data indicated that Ad5-TMEM166 may be a novel gene therapy candidate for cancer.

[Construction of recombinant NF-κB reporter adenovirus and activity analysis].

OBJECTIVE: To study the feasibility of adenovirus-based nuclear factor-κB (NF-κB) reporter as a model to screen the upstream signal regulators of NF-κB. METHODS: A type 5 (E1/E3 deficient) adenovirus vector pAdxsi was used to construct the NF-κB reporter adenovirus. Multiple adherent and suspending cell lines were infected by the NF-κB reporter adenovirus, and the luciferase activity of the NF-κB reporter gene was measured. RESULTS: An NF-κB reporter adenovirus (Ad-NF-κB-luc) was successfully constructed. The virus was capable of infecting HepG2, MGC803, THP-1 and U937 cell lines and showed high activities of NF-κB-luc reporter gene when stimulated by tumor necrosis factor-α (TNF-α) or lipopolysaccharide (LPS). CONCLUSION: The Ad-NF-κB-luc reporter gene transfer system can effectively infect those cells hard-transfected by conventional transfection reagents. It also produces a high activity of NF-κB-luc reporter gene with stability and reliability. Our study expands the application of NF-κB reporter gene.

Human HS1BP3 induces cell apoptosis and activates AP-1.

In the present study, we characterized the function of HS1-binding protein 3 (HS1BP3), which is mutated in essential tremor and may be involved in lymphocyte activation. We found that HS1BP3 localized to the mitochondria and endoplasmic reticulum partially. Overexpression of HS1BP3 induced apoptosis in HEK293T and HeLa cell lines. When these cell lines were transfected with HS1BP3, they exhibited nuclear DNA condensation, externalization of phosphatidylserine (PS), and cleavage of poly ADP ribose polymerase (PARP). Furthermore, suppression of HS1BP3 or HS1 expression attenuates HS1BP3 induced apoptosis. In addition, HS1BP3 enhanced activator protein 1 (AP-1)-mediated transcription in a dose-dependent manner. Therefore, we conclude that HS1BP3 regulates apoptosis via HS1 and stimulates AP-1-mediated transcription.

Human SBK1 is dysregulated in multiple cancers and promotes survival of ovary cancer SK-OV-3 cells.

Protein kinases are involved in comprehensive cellular processes and also implicated in many human diseases. SH3-binding domain kinase 1 (SBK1) was first cloned and characterized in rat and the human cDNA was cloned in our lab in 2006, but the expression and function of endogenous protein have not been well studied in human. In this follow up study, we screened a panel of cell lines and tissues, as well as a tumor tissue array for SBK1 expression at both RNA and protein levels. To detect the protein, we generated the first rabbit polyclonal antibody against human SBK1. We show that the SBK1 is expressed in most of the cells and tissues examined, and the protein is highly up-regulated in ovarian serous adenocarcinoma while down-regulated in esophagus squamous cell carcinoma and stomach adenocarcinoma. When over-expressed in an ovarian cancer cells SK-OV-3 by adenovirus infection, SBK1 protected the cells from apoptosis induced by the viral infection, therefore promoting cancer cell survival. Given that a missense mutation K92E in human SBK1 was identified recently from ovarian mucinous carcinoma, together, these results suggest that the wide-spread expression pattern of human SBK1 may predict a broad cellular function, and its dysregulated in certain cancers suggests an involvement of the protein in the pathogenesis of human cancers.

High-throughput cell-based screening reveals a role for ZNF131 as a repressor of ERalpha signaling.

BACKGROUND: Estrogen receptor alpha (ERalpha) is a transcription factor whose activity is affected by multiple regulatory cofactors. In an effort to identify the human genes involved in the regulation of ERalpha, we constructed a high-throughput, cell-based, functional screening platform by linking a response element (ERE) with a reporter gene. This allowed the cellular activity of ERalpha, in cells cotransfected with the candidate gene, to be quantified in the presence or absence of its cognate ligand E2. RESULTS: From a library of 570 human cDNA clones, we identified zinc finger protein 131 (ZNF131) as a repressor of ERalpha mediated transactivation. ZNF131 is a typical member of the BTB/POZ family of transcription factors, and shows both ubiquitous expression and a high degree of sequence conservation. The luciferase reporter gene assay revealed that ZNF131 inhibits ligand-dependent transactivation by ERalpha in a dose-dependent manner. Electrophoretic mobility shift assay clearly demonstrated that the interaction between ZNF131 and ERalpha interrupts or prevents ERalpha binding to the estrogen response element (ERE). In addition, ZNF131 was able to suppress the expression of pS2, an ERalpha target gene. CONCLUSION: We suggest that the functional screening platform we constructed can be applied for high-throughput genomic screening candidate ERalpha-related genes. This in turn may provide new insights into the underlying molecular mechanisms of ERalpha regulation in mammalian cells.

Identification of five human novel genes associated with cell proliferation by cell-based screening from an expressed cDNA ORF library.

The development of functional profiling technologies provides opportunity for high-throughput functional genomics studies. We describe a cell-based screening system to identify novel human genes associated with cell proliferation. The method integrates luciferase reporter gene activity, fluorescence stain, automated microscopy and cellular phenotype assays. We successfully used the system to screen 409 novel human genes cloned by our lab and found that 27 genes significantly up-regulated promoter-Renilla luciferase reporter plasmid (pRL) activity. Among them, five genes, TRAF3IP3, ZNF306, ZNF250, SGOL1, and ZNF434, were determined through morphological observation, calcein AM fluorescence stain, MTT assay and cell cycle analysis to be associated with cell proliferation. Furthermore, we showed that the gene TRAF3IP3, which initially was identified to specifically interact with TRAF3, stimulated cell growth by modulating the c-Jun N-terminal kinase (JNK) pathway, and RNAi of TRAF3IP3 confirmed that the effect was physiological and necessary. In summary, we integrated a rapid and efficient system for screening novel growth regulatory genes. Using the new screening system we identified five genes associated with cell proliferation for the first time.

TMEM166, a novel transmembrane protein, regulates cell autophagy and apoptosis.

Programmed cell death can be divided into apoptosis and autophagic cell death. We describe the biological activities of TMEM166 (transmembrane protein 166, also known as FLJ13391), which is a novel lysosome and endoplasmic reticulum-associated membrane protein containing a putative TM domain. Overexpression of TMEM166 markedly inhibited colony formation in HeLa cells. Simultaneously, typical morphological characteristics consistent with autophagy were observed by transmission electron microscopy, including extensive autophagic vacuolization and enclosure of cell organelles by double-membrane structures. Further experiments confirmed that the overexpression of TMEM166 increased the punctate distribution of MDC staining and GFP-LC3 in HeLa cells, as well as the LC3-II/LC3-I proportion. On the other hand, TMEM166-transfected HeLa and 293T cells succumbed to cell death with hallmarks of apoptosis including phosphatidylserine externalization, loss of mitochondrial transmembrane potential, caspase activation and chromatin condensation. Kinetic analysis revealed that the appearance of autophagy-related biochemical parameters preceded the nuclear changes typical of apoptosis in TMEM166-transfected HeLa cells. Suppression of TMEM166 expression by small interference RNA inhibited starvation-induced autophagy in HeLa cells. These findings show for the first time that TMEM166 is a novel regulator involved in both autophagy and apoptosis.

Screening for novel human genes associated with CRE pathway activation with cell microarray.

In this study, cell microarray technology is used to identify novel human genes associated with CRE pathway activation. By reverse transfection, expression plasmids containing full-length cDNAs were cotransfected with the reporter plasmid pCRE-d2EGFP to monitor the activation of the CRE pathway via enhanced green fluorescence protein (EGFP) expression. Of the 575 predominantly novel genes screened, 22 exhibited relatively higher EGFP fluorescence compared with a negative control. After a functional validation with a dual luciferase reporter system that included both cis- and trans-luciferase assays, 4 of the 22 genes (RNF41, C8orf32, C6orf208, and MEIS3P1) were confirmed as CRE-pathway activators. Western blot analysis revealed that RNF41 can promote CREB phosphorylation. These results demonstrate the successful combination of cell microarray technology with this reporting system and the potential of this tool to characterize functions of novel genes in a highly parallel format.