STK19 is a DNA/RNA-binding protein critical for DNA damage repair and cell proliferation.

STK19 was originally identified as a manganese-dependent serine/threonine-specific protein kinase, but its function has been highly debated. Here, the crystal structure of STK19 revealed that it does not contain a kinase domain, but three intimately packed winged helix (WH) domains. The third WH domain mediated homodimerization and double-stranded DNA binding, both being important for its nuclear localization. STK19 participated in the nucleotide excision repair (NER) and mismatch repair (MMR) pathways by recruiting damage repair factors such as RPA2 and PCNA. STK19 also bound double-stranded RNA through the DNA-binding interface and regulated the expression levels of many mRNAs. Furthermore, STK19 knockdown cells exhibited very slow cell proliferation, which cannot be rescued by dimerization or DNA-binding mutants. Therefore, this work concludes that STK19 is highly unlikely to be a kinase but a DNA/RNA-binding protein critical for DNA damage repair (DDR) and cell proliferation. To prevent further confusions, we renamed this protein as TWH19 (Tandem Winged Helix protein formerly known as STK19).

Hippocampal circAnk3 Deficiency Causes Anxiety-like Behaviors and Social Deficits by Regulating the miR-7080-3p/IQGAP1 Pathway in Mice.

BACKGROUND: Circular RNAs are highly enriched in the synapses of the mammalian brain and play important roles in neurological function by acting as molecular sponges of microRNAs. circAnk3 is derived from the 11th intron of the ankyrin-3 gene, Ank3, a strong genetic risk factor for neuropsychiatric disorders; however, the function of circAnk3 remains elusive. In this study, we investigated the function of circAnk3 and its downstream regulatory network for target genes in the hippocampus of mice. METHODS: The DNA sequence from which circAnk3 is generated was modified using CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/Cas9) technology, and neurobehavioral tests (anxiety and depression-like behaviors, social behaviors) were performed in circAnk3+/- mice. A series of molecular and biochemical assays were used to investigate the function of circAnk3 as a microRNA sponge and its downstream regulatory network for target genes. RESULTS: circAnk3+/- mice exhibited both anxiety-like behaviors and social deficits. circAnk3 was predominantly located in the cytoplasm of neuronal cells and functioned as a miR-7080-3p sponge to regulate the expression of Iqgap1. Inhibition of miR-7080-3p or restoration of Iqgap1 in the hippocampus ameliorated the behavioral deficits of circAnk3+/- mice. Furthermore, circAnk3 deficiency decreased the expression of the NMDA receptor subunit GluN2a and impaired the structural plasticity of dendritic synapses in the hippocampus. CONCLUSIONS: Our results reveal an important role of the circAnk3/miR-7080-3p/IQGAP1 axis in maintaining the structural plasticity of hippocampal synapses. circAnk3 might offer new insights into the involvement of circular RNAs in neuropsychiatric disorders.

Repression of LSD1 potentiates homologous recombination-proficient ovarian cancer to PARP inhibitors through down-regulation of BRCA1/2 and RAD51.

Poly (ADP-ribose) polymerase inhibitors (PARPi) are selectively active in ovarian cancer (OC) with homologous recombination (HR) deficiency (HRD) caused by mutations in BRCA1/2 and other DNA repair pathway members. We sought molecular targeted therapy that induce HRD in HR-proficient cells to induce synthetic lethality with PARPi and extend the utility of PARPi. Here, we demonstrate that lysine-specific demethylase 1 (LSD1) is an important regulator for OC. Importantly, genetic depletion or pharmacological inhibition of LSD1 induces HRD and sensitizes HR-proficient OC cells to PARPi in vitro and in multiple in vivo models. Mechanistically, LSD1 inhibition directly impairs transcription of BRCA1/2 and RAD51, three genes essential for HR, dependently of its canonical demethylase function. Collectively, our work indicates combination with LSD1 inhibitor could greatly expand the utility of PARPi to patients with HR-proficient tumor, warranting assessment in human clinical trials.

Discovery and Optimization of Novel hDHODH Inhibitors for the Treatment of Inflammatory Bowel Disease.

As a key rate-limiting enzyme in the de novo synthesis of pyrimidine nucleotides, human dihydroorotate dehydrogenase (hDHODH) is considered a known target for the treatment of autoimmune diseases, including inflammatory bowel disease (IBD). Herein, BAY 41-2272 with a 1H-pyrazolo[3,4-b]pyridine scaffold was identified as an hDHODH inhibitor by screening an active compound library containing 5091 molecules. Further optimization led to 2-(1-(2-chloro-6-fluorobenzyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-cyclopropylpyrimidin-4-amine (w2), which was found to be the most promising and drug-like compound with potent inhibitory activity against hDHODH (IC50 = 173.4 nM). Compound w2 demonstrated acceptable pharmacokinetic characteristics and alleviated the severity of acute ulcerative colitis induced by dextran sulfate sodium in a dose-dependent manner. Notably, w2 exerted better therapeutic effects on ulcerative colitis than hDHODH inhibitor vidofludimus and Janus kinase (JAK) inhibitor tofacitinib. Taken together, w2 is a promising hDHODH inhibitor for the treatment of IBD and deserves to be developed as a preclinical candidate.

Neonatal exposure to sevoflurane induces adolescent neurobehavioral dysfunction by interfering with hippocampal glycerophoslipid metabolism in rats.

Sevoflurane exposure in the neonatal period causes long-term developmental neuropsychological dysfunction, including memory impairment and anxiety-like behaviors. However, the molecular mechanisms underlying such effects have not been fully elucidated. In this study, we investigated the effect of neonatal exposure to sevoflurane on neurobehavioral profiles in adolescent rats, and applied an integrated approach of lipidomics and proteomics to investigate the molecular network implicated in neurobehavioral dysfunction. We found that neonatal exposure to sevoflurane caused cognitive impairment and social behavior deficits in adolescent rats. Lipidomics analyses revealed that sevoflurane significantly remodeled hippocampal lipid metabolism, including lysophatidylcholine (LPC) metabolism, phospholipid carbon chain length and carbon chain saturation. Through a combined proteomics analysis, we found that neonatal exposure to sevoflurane significantly downregulated the expression of lysophosphatidylcholine acyltransferase 1 (LPCAT1), a key enzyme in the regulation of phospholipid metabolism, in the hippocampus of adolescent rats. Importantly, hippocampal LPCAT1 overexpression restored the dysregulated glycerophospholipid (GP) metabolism and alleviated the learning and memory deficits caused by sevoflurane. Collectively, our evidence that neonatal exposure to sevoflurane downregulates LPCAT1 expression and dysregulates GP metabolism in the hippocampus, which may contribute to the neurobehavioral dysfunction in the adolescent rats.

[Modified internal fixator combined with sacroiliac screws in treatment of Tile C1.3 pelvic fracture].

Objective: To investigate the effectiveness of modified internal fixator (INFIX) to fix the anterior pelvic ring for Tile C1.3 pelvic fracture by comparing with the traditional INFIX. Methods: The patients with Tile C1.3 pelvic fractures admitted between April 2018 and June 2021 were taken as the study objects. Of them, 55 cases were included in the study according to the selection criteria. During operation, the anterior pelvic ring was fixed with the modified INFIX in 30 cases (modified group) and with the traditional INFIX in 25 cases (control group). The posterior pelvic ring in 55 cases was fixed with sacroiliac screws. There was no significant difference between the two groups in gender, age, cause of injury, time from injury to operation, and combined injury ( P>0.05). The operation time, intraoperative blood loss, and complications were recorded and compared between the two groups. The X-ray film was taken to evaluate the reduction of the anterior pelvic ring after operation, grade the reduction according to the Matta scoring standards, and observe the fracture healing and healing time. The function was evaluated according to Majeed scoring standards during follow-up. Results: The operation time of the modified group was significantly longer than that of the control group ( Z=-3.837, P<0.001), but there was no significant difference in the intraoperative blood loss between the two groups ( t=-1.831, P=0.076). All patients were followed up 12-18 months (mean, 14 months). X-ray film reexamination showed that the excellent and good rate of Matta scoring for anterior pelvic ring reduction in the modified group was 88.00%, which was significantly better than that in the control group (63.33%) ( χ 2=4.373, P=0.037). All fractures healed, and the fracture healing time of the modified group and the control group was (12.04±3.59) and (14.83±4.83) weeks respectively, with a significant difference ( t=2.401, P=0.020). At last follow-up, the excellent and good rates of Majeed scoring were 80.00% in the modified group and 76.67% in the control group, with no significant difference ( χ 2=0.089, P=0.766). In the modified group, 2 cases (8%) had complications, all of which were incision infection above pubic symphysis. In the control group, 9 cases (30%) had complications, including 5 cases of lateral femoral cutaneous nerve injury, 2 cases of femoral nerve paralysis, and 2 cases of delayed fracture healing. The incidence of complications was significantly lower in the modified group than in the control group ( χ 2=4.125, P=0.042). Conclusion: Compared with the traditional INFIX, the modified INFIX to fix the anterior pelvic ring for Tile C1.3 pelvic fracture has fewer complications, better stability, shorter fracture healing time, and lower risk of nerve injury.

The Src-Kinase Fyn is Required for Cocaine-Associated Memory Through Regulation of Tau.

Drug-associated context-induced relapse of cocaine-seeking behaviour requires the retrieval of drug-associated memory. Studies exploring the underlying neurobiological mechanism of drug memory formation will likely contribute to the development of treatments for drug addiction and the prevention of relapse. In our study, we applied a cocaine-conditioned place preference (CPP) paradigm and a self-administration paradigm (two drug-associated memory formation model) to confirm the hypothesis that the Src kinase Fyn critically regulates cocaine-associated memory formation in the hippocampus. For this experiment, we administered the Src kinase inhibitor PP2 into the bilateral hippocampus before cocaine-CPP and self-administration training, and the results showed that pharmacological manipulation of the Src kinase Fyn activity significantly attenuated the response to cocaine-paired cues in the cocaine-CPP and self-administration paradigms, indicating that hippocampal Fyn activity contributes to cocaine-associated memory formation. In addition, the regulation of cocaine-associated memory formation by Fyn depends on Tau expression, as restoring Tau to normal levels disrupted cocaine memory formation. Together, these results indicate that hippocampal Fyn activity plays a key role in the formation of cocaine-associated memory, which underlies cocaine-associated contextual stimulus-mediated regulation of cocaine-seeking behaviour, suggesting that Fyn represents a promising therapeutic target for weakening cocaine-related memory and treating cocaine addiction.

[Current status and progress of locking plate in the treatment of distal femoral comminuted fracture].

OBJECTIVE: To review the current status and progress of locking plate for the treatment of distal femoral comminuted fractures. METHODS: The related literature was extensively reviewed to summarize the current status and progress in the treatment of distal femoral comminuted fracture with locking plate from four aspects: the current treatment situation, the shortcomings of locking plate and countermeasures, the progress of locking technology, locking plate and digital orthopedic technology. RESULTS: Treatment of distal femoral comminuted fractures is challenging. Locking plates, the most commonly used fixation for distal femoral comminuted fractures, still face a high rate of treatment failure. Double plates can improve the mechanical stability of comminuted fractures, but specific quantitative criteria are still lacking for when to choose double plates for fixation. The far cortial locking screw has shown good application value in improving the micro-movement and promoting the growth of callus. The biphasic plating is a development of the traditional locking plate, but needs further clinical examination. As an auxiliary means, digital orthopedic technology shows a good application prospect. CONCLUSION: The inherent defect of locking plate is a factor that affects the prognosis of distal femoral comminuted fracture. The optimization of locking technology combined with digital orthopedic technology is expected to reduce the failure rate of treatment of distal femoral comminuted fracture.

[Research on the nature of micromovement and the biomechanical staging of fracture healing].

OBJECTIVE: To explore the nature of micromovement and the biomechanical staging of fracture healing. METHODS: Through literature review and theoretical analysis, the difference in micromovement research was taken as the breakthrough point to try to provide a new understanding of the role of micromovement and the mechanical working mode in the process of fracture healing. RESULTS: The process of fracture healing is the process of callus generation and connection. The micromovement is the key to start the growth of callus, and the total amount of callus should be matched with the size of the fracture space. The strain at the fracture end is the key to determine the callus connection. The strain that can be tolerated by different tissues in the fracture healing process will limit the micromovement. According to this, the fracture healing process can be divided into the initiation period, perfusion period, contradiction period, connection period, and physiological period, i.e., the biomechanical staging of fracture healing. CONCLUSION: Biomechanical staging of fracture healing incorporates important mechanical parameters affecting fracture healing and introduces the concepts of time and space, which helps to understand the role of biomechanics, and its significance needs further clinical test and exploration.

Clinical Results of Acetabular Fracture via the Pararectus versus Ilioinguinal Approach.

OBJECTIVE: To compare the clinical efficacy of pararectus and ilioinguinal approach in the treatment of acetabular fractures. METHODS: A retrospective analysis of the clinical data of 60 patients with acetabular fractures treated by the pararectus approach or the ilioinguinal approach from January 2016 to January 2019 was performed to record all data by comparing the length of the surgical incision, the time to expose the fracture and the amount of blood loss during the operation. Patients were routinely followed up at 1, 6 and 12 months postoperatively. The function of the hip joint after the operation (Improved Merle d' Aubigne and Postel scores) and the complications postoperation were recorded. RESULTS: There was a significant difference (mean ± SD) in the length of surgical incision [(11.2 ± 1.5) cm vs.(23.8 ± 2.1) cm], and in surgical exposure time [(10.8 ± 1.7) min vs.(19.9 ± 1.9) min] (P < 0.05) between the two approaches; there was no significant difference (mean ± SD) in intraoperative blood loss [(591.8 ± 131.4) mL vs. (614.6 ± 132.7) mL] or in hip function scores at the last follow-up between the two groups (P > 0.05). In the pararectus approach group, there was one patient (3.3%) with postoperative wound fat liquefaction, and the wound completely improved by secretion culture, enhanced dressing and effective antibiotics, one patient (3.3%) developed lateral femoral cutaneous nerve injury; One case (3.3%) of postoperative myositis ossificans occurred in the ilioinguinal approach group, and there were no obvious symptoms. CONCLUSIONS: These data suggest that for patients with acetabular fractures, both the pararectus approach and the ilioinguinal approach can achieve satisfactory surgical results, but the former has relatively simple operation and small incision length, which is in line with the modern concept of the minimally invasive pelvis.

Cytoplasmic SHMT2 drives the progression and metastasis of colorectal cancer by inhibiting ß-catenin degradation.

Introduction: Serine hydroxymethyltransferase 2 (SHMT2) plays a critical role in serine-glycine metabolism to drive cancer cell proliferation. However, the nonmetabolic function of SHMT2 in tumorigenesis, especially in human colorectal cancer (CRC) progression, remains largely unclear. Methods: SHMT2 expression in human CRC cells was identified by western blot and immunofluorescence assay. The CRC cell proliferation, migration, and invasion after SHMT2 knockdown or overexpression were explored through in vitro and in vivo assays. Immunofluorescence, mRNA-seq, co-immunoprecipitation, chromatin immunoprecipitation-qPCR and immunohistochemistry assays were used to investigate the underlying mechanisms behind the SHMT2 nonmetabolic function. Results: We demonstrated that SHMT2 was distributed in the cytoplasm and nucleus of human CRC cells. SHMT2 knockdown resulted in the significant inhibition of CRC cell proliferation, which was not restored by serine, glycine, or formate supplementation. The invasion and migration of CRC cells were suppressed after SHMT2 knockdown. Mechanistically, SHMT2 interacted with ß-catenin in the cytoplasm. This interaction inhibited the ubiquitylation-mediated degradation of ß-catenin and subsequently modulated the expression of its target genes, leading to the promotion of CRC cell proliferation and metastasis. Notably, the lysine 64 residue on SHMT2 (SHMT2K64) mediated its interaction with ß-catenin. Moreover, transcription factor TCF4 interacted with ß-catenin, which in turn increased SHMT2 expression, forming an SHMT2/ß-catenin positive feedback loop. In vivo xenograft experiments confirmed that SHMT2 promoted the growth and metastasis of CRC cells. Finally, the level of SHMT2 was found to be significantly increased in human CRC tissues. The SHMT2 level was correlated with an increased level of ß-catenin, associated with CRC progression and predicted poor patient survival. Conclusion: Taken together, our findings reveal a novel nonmetabolic function of SHMT2 in which it stabilizes ß-catenin to prevent its ubiquitylation-mediated degradation and provide a potential therapeutic strategy for CRC therapy.

Therapeutic Effect of Acetabular Fractures Using the Pararectus Approach Combined with 3D Printing Technique.

OBJECTIVE: To explore the clinical efficacy of pararectus approach combined with 3D printing technique for the surgical treatment of partial acetabular fractures. METHODS: We retrospectively evaluated 33 (20 males and 13 females) patients with acetabular fractures in the period of June 2017 to December 2018. According to Judet and Letournel classification: 11 cases were of anterior column fracture, 10 cases were of double column fracture, seven fractures were of the anterior column with posterior half transverse, three fractures were of transverse fracture, and two cases were of "T" fracture. For all cases, 3D printing is used to print the acetabular model. Pre-bent reconstruction plates from the model were placed to fixate fractures via the pararectus approach. RESULTS: Thirty-three patients (mean age 48 years; range, 35-63 years), included 20 men and 13 women, were treated successfully with open reduction and internal fixation by the pararectus approach. Surgery duration was 203 min on average (range: 135-245 min), and intra-operative bleeding was 1030 mL on average (range: 450-1400 mL). All patients were followed-up for 12-18 months (average,14 months); two patients (6.0%) developed postoperative ossifying myositis, and there are no obvious symptoms at present; one patient (3.0%) developed postoperative wound infection, and the wound completely improved by secretion culture, enhanced dressing, and effective antibiotics; all the acetabular fractures united after 12 to 16 weeks (average,13 weeks). According to the modified Merle d'Aubigne and Postel scoring system to assess the hip function: excellent in 22 cases (66.7%), good in seven cases (21.2%), and fair in four cases (12.1%). CONCLUSIONS: In the treatment of partial acetabular fractures, the pararectus approach combined with 3D printing technique can achieve effective reduction and fixation, decrease intraoperative hemorrhage, shorten operation time, and the internal fixation position can be properly adjusted during the operation by looking directly at the model.

[Early effectiveness analysis of lateral hinge fracture during medial opening-wedge high tibial osteotomy].

OBJECTIVE: To compare the clinical and radiologic effectiveness in patients with versus without lateral hinge fracture during medial opening-wedge high tibial osteotomy (MOWHTO) to evaluate the effect of lateral hinge fracture on short-term effectiveness. METHODS: The clinical data of 84 patients (97 knees) with medial compartment osteoarthritis who treated with MOWHTO between September 2015 and July 2018 was retrospectively analyzed. There were 10 males (10 knees) and 74 females (87 knees). The age ranged from 45 to 65 years with an average of 57.7 years. Lateral hinge fracture was recognized by the intraoperative fluoroscopy or immediate postoperative X-ray film. Fractures were classified into types Ⅰ, Ⅱ, and Ⅲ according to the Takeuchi classification. The healing of osteotomy was observed by radiographs during follow-up; the femur tibia angle (FTA), medialproximal tibial angle (MPTA), and hip-knee-ankle angle (HKA) were also calculated. The knee joint function was evaluated by Hospital for Special Surgery (HSS) score and knee society score (KSS). RESULTS: The incision healed by first intention. All patients were followed up 15-48 months with an average of 24.8 months. No hinge fracture occurred in 78 knees (80.41%, group A), and lateral hinge fractures were observed in 19 knees (19.59%, group B) and were divided into the type Ⅰ (13 knees, 13.40%) and type Ⅲ (6 knees, 6.19%) groups. Type Ⅰ fractures were not additionally treated, type Ⅲ fractures were anatomic reduced fixed with additional lag screws. X-ray film and CT examination showed that all patients had bone healing at 3 months after operation without delayed healing or nonunion. During follow-up, there was no loosening or fracture of internal fixation plates and screws. HKA, FTA, and MPTA of patients in group A and group B (type Ⅰ and Ⅲ) were significantly improved at each time point after operation compared with preoperative values ( P<0.05); there was no significant difference between groups at each time point before and after operation ( P>0.05). After operation, the pain of knee joint was alleviated and the function of joint was improved. At last follow-up, KSS score and HSS score of groups A and B were significantly improved compared with those before operation ( P<0.05), but there was no significant difference between the two groups ( P>0.05). CONCLUSION: The lateral hinge fracture may occur during MOWHTO. As long as the treatment and rehabilitation were guided according to the fracture classification of the hinge, the effectiveness can be similar to those without the hinge fracture.

Isolation and genomic characterization of a cypovirus from the oleander hawk moth, Daphnis nerii.

The oleander hawk moth, Daphnis nerii, is a serious pest of plants belonging to the family Apocynaceae. Thus far, pathogen infection has not been reported in D. nerii. In this study, a new cytoplasmic polyhedrosis virus (cypovirus; CPV) was isolated from naturally diseased D. nerii larvae and named DnCPV-23. Virions were observed in ultrathin sections of DnCPV polyhedral bodies. Electrophoretic analysis revealed that the DnCPV genome consisted of 10 segments of double-stranded RNA (dsRNA). cDNA copies of these dsRNA segments were amplified using the method of full-length amplification of cDNAs (FLAC), cloned, and sequenced. Sequencing results showed that all segments contained one open reading frame (ORF); They shared the conserved terminal sequences AGUCAAA and AGC at 5' and 3' ends respectively, except segment 4, which is different from previously reported 22 cypoviruses. Phylogenetic analysis based on amino acid sequences of polyhedrin (encoded by segment 10) indicated that this CPV was closely related to CPV type 19. Altogether, DnCPV-23 is a new type of cypovirus.

[Application of three-dimensional printing technology for closed reduction and percutaneous cannulated screws fixation of displaced intraarticular calcaneus fractures].

Objective: To evaluate the effectiveness of three-dimensional (3-D) printing assisting minimally invasive for intraarticular calcaneal fractures with percutaneous poking reduction and cannulate screw fixation. Methods: A retrospective analysis was performed of the 19 patients (19 feet) with intraarticular calcaneal fracture who had been treated between March 2015 and May 2016. There were 13 males and 6 females with an average age of 38.2 years (range, 24-73 years). There were 3 open fractures and 16 closed fractures. By Sanders classification, 12 cases were type Ⅱ, 7 cases were type Ⅲ. By Essex-Lopresti classification, 13 cases were tongue type, 6 cases were joint-depression type. The time from injury to surgery was 1-10 days (mean, 4.7 days). A thin slice CT scan was taken of bilateral calcaneus in patients. By using the mirror imaging technique, the contralateral mirror image and the affected side calcaneus model were printed according to 1∶1 ratio. The displacement of fracture block was observed and contrasted, and the poking reduction was simulated. Calcaneal fracture was treated by percutaneous minimally invasive poking reduction and cannulate screw fixation. The Böhler angle and Gissane angle at immediate after operation and last follow-up was measured on X-ray films, and compared with preoperative measurement. The functional recovery was evaluated by American Orthopaedic Foot and Ankle Society (AOFAS) scores. Results: The operation time was 25-70 minutes (mean, 45 minutes). The intraoperative blood loss was 10-40 mL (mean, 14.5 mL). All the incisions healed by first intention and had no relevant postoperative complications such as skin necrosis, nail tract infection, and osteomyelitis. All the patients were followed up 12-25 months (mean, 14.6 months). All patients obtained fracture healing, and the fracture healing time was 8-14 weeks (mean, 10.3 weeks). No screw withdrawal or breakage occurred during follow-up; only 1 patient with Sanders type Ⅱ fracture, whose calcaneus height was partially lost at 6 weeks after operation, the other patients had no reduction loss and fracture displacement, and no traumatic arthritis occurred. The Böhler angle and Gissane angle at immediate after operation and last follow-up were significantly improved when compared with preoperative ones ( P<0.05), but there was no significant difference between at immediate after operation and last follow-up ( P>0.05). The AOFAS score was 76-100 (mean, 88.2), and the results were excellent in 10 feet, good in 7, and fair in 2, the excellent and good rate was 89.5%. Conclusion: 3-D printing assisting minimally invasive for intraarticular calcaneal fractures with percutaneous poking reduction and cannulate screw fixation can reduce the surgical trauma, improve the quality of reduction and fixation, and make the operation more safe, accurate, and individualized.

[Effectiveness of locking compress plate and extra cortical bone bridge fixation for treatment of atrophic humeral nonunions].

Objective: To assess the effectiveness of locking compress plate and extra cortical bone bridge fixation for treating atrophic humeral nonunion. Methods: Seventeen patients with atrophic humeral nonuninon were treated with locking compress plate and extra cortical bone bridge fixation between November 2006 and June 2015. Of 17 cases, 11 were male, 6 were female, aged 24-63 years (mean, 38.2 years). Fracture located at the left side in 9 cases and at the right side in 8 cases. The mechanism of injury was traffic accident in 13 cases, falling from height in 3 cases, and heavy pound injury in 1 case. The patients underwent surgery for 1 time in 7 cases, for 2 times in 5 cases, for 3 times in 4 cases, and for 4 times in 1 case. The time from fracture to hospitalization was 10-76 months (mean, 22.6 months). The shoulder function was evaluated by Neer score, and elbow function by Mayo score. Results: All incisions healed by first intention. Two cases had transient radial nerve symptoms of numbness. All patients were followed up 27.3 months on average (range, 15-60 months). Radiographic examination showed signs of bone remodeling at 6-8 weeks after operation, and formation of extra cortical bone bridge. All of them achieved bone union within 10 to 41 weeks (mean, 17.6 weeks). At last follow-up, the average Neer score was 83.36 (range, 72-96); and the shoulder function was excellent in 10 cases, good in 5, and fair in 2 with an excellent and good rate of 88.24%. And the average Mayo score was 86.52 (range, 68-100); and the elbow function was excellent in 11 cases, good in 3, and fair in 3 with an excellent and good rate of 82.35%. Conclusion: The bone bridging could effectively form by extra cortical grafting technique. Atrophic humeral nonunions can be successfully treated with locking compress plate and extra cortical bone bridge fixation.

Over-expression of Sox4 and ß-catenin is associated with a less favorable prognosis of osteosarcoma.

The purpose of this study was to examine the association of the expression of Sox4 and ß-catenin with the prognosis of osteosarcoma. A total of 108 cases of conventional osteosarcoma were involved in this study and 28 cases of osteochondroma served as controls. The expression of Sox4 and ß-catenin was detected by using immunohistochemical staining and Western blotting. The results showed that Sox4 and ß-catenin were over-expressed in 67 (62.03%) and 62 (57.41%) of 108 osteosarcoma cases, while in only 3 (10.71%) and 5 (17.86%) of 28 controls, respectively (P<0.05 for all). The expression of Sox4 and ß-catenin was associated with the distant metastasis, pathological grade and Enneking stage of patients with osteosarcoma (P<0.05 for all). The mean overall survival time and the 5-year-survival rate in osteosarcoma patients with Sox4 and ß-catenin over-expressed were significantly reduced as compared with those in Sox4 and ß-catenin low-expression group (P<0.05 for all). Cox multifactor regression analysis revealed that the distant metastasis, Enneking stage, and the expression of Sox4 and ß-catenin were independent risk factors of patients with osteosarcoma (P<0.05 for all). The findings indicated that overexpression of Sox4 and ß-catenin is associated with a poor prognosis of osteosarcoma.

[Isolation and Identification of a Novel Cypovirus from Daphnis nerii].

In order to develop a novel effective biological insecticide for controlling oleander hawk moth, a new pathogen was isolated from naturally diseased Daphnis nerii. Based on scanning electron microscopy, full-length amplification of cDNAs (FLAC), and phylogenetic analysis of genome segments 2and 10,the virus was identified as a new type of cypovirus (Da phnis nerii cypovirus [DnCPV]). Electrophoresis analysis showed that DnCPV had a genome comprising 10double-stranded RNA (dsRNA) segments, ranging from 892 to 4160bp.Using FLAC, the cDNAs from the 10 dsRNA segments of the new CPV were cloned and genome segments 2and 10 were sequenced. Sequencing results showed that segment 2 encoded RNA-dependent-RNA-polymerases (RdRps) and segment 10 encoded polyhedrin. These two segments shared conserved terminal sequences of AGUCAAA and AGC at the 5'and 3'ends,respectively.These conserved terminal sequences were not consistent with any of the known CPV types.Phylogenetic analysis of the RdRp and polyhedrin indicated that this CPV was more closely related to CPV type 19 and type 5than other CPV types. Based on the unique conserved terminal sequences and the electrophoresis pattern of the new virus, we tentatively named it DnCPV Nanchang isolate: DnCPV-NC.

[LONG-TERM EFFECTIVENESS OF TRANSPIANTATION OF ILIAC BONE FLAP PEDICLED WITH DEEP ILIAC CIRCUMFLEX VESSELS FOR AVASCULAR NECROSIS OF FEMORAL HEAD AT STAGE II AND III].

OBJECTIVE: To assess the long-term effectiveness of the transplantation of iliac bone flap pedicled with deep iliac circumflex vessels for treating avascular necrosis of femoral head (ANFH) at Ficat stage II and III. METHODS: Thirty-two cases (43 hips) of ANFH underwent iliac bone flap transplantation pedicled with deep iliac circumflex vessels between October 2000 and February 2006, and the clinical data were retrospectively reviewed. Of 32 cases, 27 were male (38 hips), and 5 were female (5 hips), aged 21-52 years (mean, 36.6 years); there were 8 cases (11 hips) of hormone ANFH, 18 cases (23 hips) of alcoholic ANFH, and 6 cases (9 hips) of idiopathic ANFH. The disease duration ranged from 2-52 months (mean, 8.2 months). According to Ficat staging criteria, 26 hips were classified as stage II and 17 hips as stage III. The preoperative Harris hip score (HSS) was 68.2±8.4. The HHS scores and X-ray photograph were compared between at pre- and post-operation to assess the outcomes clinically and radiologically. RESULTS: All incisions healed by first intention. Two cases had numbness of the lateral femoral skin. Four patients (6 hips) failed to be followed up, and the other 28 cases were followed up 98-187 months (mean, 129.3 months). Five patients (6 hips) showed aggravation or no relief with progression to stage IV at 8-69 months, who received total hip arthroplasty. The 10-year survival rate was 83.78% (31/37). The HHS score was significantly increased to 86.7±9.0 at last follow-up (t=-48.313, P=0.000). The hip function was excellent in 9 hips, good in 13 hips, and fair in 9 hips, and the success rate was 75.68%. Radiographic examination showed signs of bone remodeling at 6-8 weeks. After bone healing, the femoral head density gradually became uniform. Until last follow-up, ANFH progressed from Ficat stage II to III in 5 hips, from Ficat stage II to IV in 3 hips, and from Ficat stage III to IV in 3 hips; complete hip shape, continuous Shenton line, and normal joint space were observed in the other 26 hips, and the radiographic success rate was 70.27% (26/37). CONCLUSIONS: The pedicled iliac bone flap transplantation for ANFH at Ficat stage II and III can provide good osteogenesis and vascular reconstruction of the femoral head. The long-term follow-up effectiveness is satisfactory.

[AN EXPERIMENTAL STUDY ON REPAIR OF SCIATIC NERVE INJURY BY Schwann-LIKE CELLS DERIVED FROM UMBILICAL CORD BLOOD MESENCHYMAL STEM CELLS].

OBJECTIVE: To evaluate the effect of using Schwann-like cells derived from human umbilical cord blood mesenchymal stem cells (hUCBMSCs) as the seed cells to repair large sciatic nerve defect in rats so as to provide the experimental evidence for clinical application of hUCBMSCs. METHODS: Fourty-five male Sprague Dawley (SD) rats in SPF grade, weighing 200-250 g, were selected. The hUCBMSCs were harvested and cultured from umbilical cord blood using lymphocyte separating and high molecular weight hydroxyethyl starch, and then was identified. The hUCBMSCs of 3rd generation were induced to Schwann-like cells, and then was identified by chemical derivatization combined with cytokine. The acellular nerve basal membrane conduit was prepared as scaffold material by the sciatic nerve of SD rats through repeated freezing, thawing, and washing. The tissue engineered nerve was prepared after 7 days of culturing Schwann-like cells (1 x 10(7) cells/mL) on the acellular nerve basal membrane conduit using the multi-point injection. The 15 mm sciatic nerve defect model was established in 30 male SD rats, which were randomly divided into 3 groups (10 rats each group). Defect was repaired with tissue engineered nerve in group A, with acellular nerve basal membrane conduit in group B, and with autologous sciatic nerve in group C. The nerve repair was evaluated through general observation, sciatic function index (SFI), nerve electrophysiology, weight of gastrocnemius muscle, and Masson staining after operation. RESULTS: The hUCBMSCs showed higher expression of surface markers of mesenchymal stem cells, and Schwann-like cells showed positive expression of glia cell specific markers such as S100b, glial fibrillary acidic protein, and P75. At 8 weeks after operation, the acellular nerve basal membrane conduit had no necrosis and liquefaction, with mild adhesion, soft texture, and good continuity at nerve anastomosis site in group A; group B had similar appearance to group A; adhesion of group C was milder than that of groups A and B, with smooth anastomotic stoma and no enlargement, and the color was similar to that of normal nerve. SFI were gradually decreased, group C was significantly greater than groups A and B, group A was significantly greater than group B (P < 0.05). The compound action potential could be detected in anastomotic site of 3 groups, group C was significantly greater than groups A and B, and group A was significantly greater than group B in amplitude and conduction velocity (P < 0.05). Atrophy was observed in the gastrocnemius of 3 groups; wet weight's recovery rate of the gastrocnemius of group C was significantly greater than that of groups A and B, and group A was significantly greater than group B (P < 0.05). Masson staining showed that large nerve fibers regeneration was found in group A, which had dense and neat arrangement with similar fiber diameter. The density and diameter of medullated fibers, thickness of myelinated axon, and axon diameter of group C were significantly greater than those of groups A and B, and group A was significantly greater than group B (P < 0.05). CONCLUSION: Tissue engineered nerves from hUCBMSCs-derived Schwann-like cells can effectively repair large defects of the sciatic nerve. hUCBMSCs-derived Schwann-like cells can be used as a source of seed cells in nerve tissue engineering.