Neuronal ceroid lipofuscinosis with DNAJC5/CSPα mutation has PPT1 pathology and exhibit aberrant protein palmitoylation.

Neuronal ceroid lipofuscinoses (NCL) are a group of inherited neurodegenerative disorders with lysosomal pathology (CLN1-14). Recently, mutations in the DNAJC5/CLN4 gene, which encodes the presynaptic co-chaperone CSPα were shown to cause autosomal-dominant NCL. Although 14 NCL genes have been identified, it is unknown if they act in common disease pathways. Here we show that two disease-associated proteins, CSPα and the depalmitoylating enzyme palmitoyl-protein thioesterase 1 (PPT1/CLN1) are biochemically linked. We find that in DNAJC5/CLN4 patient brains, PPT1 is massively increased and mis-localized. Surprisingly, the specific enzymatic activity of PPT1 is dramatically reduced. Notably, we demonstrate that CSPα is depalmitoylated by PPT1 and hence its substrate. To determine the consequences of PPT1 accumulation, we compared the palmitomes from control and DNAJC5/CLN4 patient brains by quantitative proteomics. We discovered global changes in protein palmitoylation, mainly involving lysosomal and synaptic proteins. Our findings establish a functional link between two forms of NCL and serve as a springboard for investigations of NCL disease pathways.

Oligomerization of Cysteine String Protein alpha mutants causing adult neuronal ceroid lipofuscinosis.

Cysteine String Protein alpha (CSPα) is a palmitoylated, synaptic vesicle co-chaperone that is essential for neuroprotection. Two mutations in CSPα - L115R and L116Δ - cause adult neuronal ceroid lipofuscinosis (ANCL), a dominantly-inherited neurodegenerative disease. To elucidate the pathogenesis of ANCL, the intrinsic biochemical properties of human wildtype (WT) and disease mutant CSPα were examined. Mutant proteins purified from Escherichia coli exhibited high potency to oligomerize in a concentration, temperature, and time dependent manner, with L115R possessing the greatest potency. When freshly purified, ANCL mutant proteins displayed normal co-chaperone activity and substrate recognition similar to WT. However, co-chaperone activity was impaired for both CSPα mutants upon oligomerization. When WT and mutant CSPα were mixed together they co-oligomerized leading to an overall decrease of co-chaperone activity. The oligomerization properties of ANCL mutants were faithfully replicated in HEK 293T cells. Interestingly, the oligomers were covalently tagged by ubiquitination instead of palmitoylation. Taken together, ANCL mutations result in both a gain and partial loss-of-function.

Porous titanium-6 aluminum-4 vanadium cage has better osseointegration and less micromotion than a poly-ether-ether-ketone cage in sheep vertebral fusion.

Interbody fusion cages made of poly-ether-ether-ketone (PEEK) have been widely used in clinics for spinal disorders treatment; however, they do not integrate well with surrounding bone tissue. Ti-6Al-4V (Ti) has demonstrated greater osteoconductivity than PEEK, but the traditional Ti cage is generally limited by its much greater elastic modulus (110 GPa) than natural bone (0.05-30 GPa). In this study, we developed a porous Ti cage using electron beam melting (EBM) technique to reduce its elastic modulus and compared its spinal fusion efficacy with a PEEK cage in a preclinical sheep anterior cervical fusion model. A porous Ti cage possesses a fully interconnected porous structure (porosity: 68 ± 5.3%; pore size: 710 ± 42 µm) and a similar Young's modulus as natural bone (2.5 ± 0.2 GPa). When implanted in vivo, the porous Ti cage promoted fast bone ingrowth, achieving similar bone volume fraction at 6 months as the PEEK cage without autograft transplantation. Moreover, it promoted better osteointegration with higher degree (2-10x) of bone-material binding, demonstrated by histomorphometrical analysis, and significantly higher mechanical stability (P < 0.01), shown by biomechanical testing. The porous Ti cage fabricated by EBM could achieve fast bone ingrowth. In addition, it had better osseointegration and superior mechanical stability than the conventional PEEK cage, demonstrating great potential for clinical application.

Resection for malignant tumors in the elbow and individualized reconstruction under assistance of 3D printing technology: A case report.

RATIONALE: With a high failure rate and multiple postoperative complications, the resection for tumors in the elbow and reconstruction present a formidable challenge to orthopedic surgeons. The maturation of 3-dimension (3D) printing technology has facilitated the preoperative design, intraoperative navigation, and reconstruction of bone defects in patients with complex malignant tumors of the elbow joint. In order to improve prognosis, we explored a method of tumor resection and elbow reconstruction aided by 3D printing technology in this research. PATIENT CONCERNS: The patient underwent nephrectomy for clear cell carcinoma of the left kidney 3 years ago. Six months ago, the patient presented with limited movement and lateral tenderness in the right elbow joint. The tumor puncture biopsy demonstrated renal clear cell carcinoma metastasis. DIAGNOSES: Renal clear cell carcinoma with distal humerus bone metastasis. INTERVENTIONS: Thin-layer CT scan data of the patient was acquired, and a 3D reconstruction of both upper limb bones and joints was conducted, followed by a simulation of diseased tissue excision. According to the model, individualized osteotomy guidelines and elbow prostheses were designed and manufactured. Then, prior to the completion of the actual operation, a simulation of the preoperative phase was performed. OUTCOMES: The operation was completed without incident. At the 1-, 3-, and 6-month postoperative examinations, both the position and mobility of the prosthesis were found to be satisfactory, and no complications were observed. The hospital for special surgery score and mayo elbow performance score scores increased in comparison to the preoperative period. LESSONS: For patients with complex tumors in the elbow joint, 3D printing technology may assist in the precise excision of the tumor and provide an individualized elbow joint prosthesis that is more precise and effective than traditional surgery. It can accomplish a satisfactory treatment effect for patients when combined with early postoperative scientific rehabilitation training, so it is a method worth promoting.

Development of a clinically relevant animal model for the talar osteonecrosis in sheep.

There are a lot of reports and reviews about osteonecrosis of the talus (ONT), yet reports about the animal model of ONT to evaluate proper therapeutic approaches are rarely heard. In our study, a novel animal model was established. Pure ethanol was injected into the cancellous bone of sheep's talus. Macroscopic observation, X-ray, CT and histology were performed at two, four, 12 and 24 weeks postoperatively. It was revealed that the trabeculae of talar head began to change their structure after two weeks postoperatively compared to the normal talus. The ONT was obvious at the end of the fourth week, and their outstanding feature was the damage of trabeculae bone and formation of cavities. CT scans and pathological changes of the subjects all showed characteristics of the early stage of osteonecrosis, also the sections of the specimens confirmed necrosis of tali. By 12 weeks, the phenomenon of necrosis still existed but fibrous tissue proliferated prominently and bone reconstruction appeared in certain area. Most specimens (3/4) got late stage necrosis which presented as synarthrosis in X-ray and mass proliferation of fibrous tissue in histology at the end of 24 weeks. The novel animal model of ONT was successful, and it is inclined to deteriorate without any intervention. The study provides us a new way to evaluate various treatments on ONT in laboratory, which may eventually pave way to clinical applications.

Knockdown of lncRNA ANRIL suppresses the production of inflammatory cytokines and mucin 5AC in nasal epithelial cells via the miR-15a-5p/JAK2 axis.

The incidence of allergic rhinitis (AR) is increasing worldwide. Human nasal epithelial cells (HNECs) are the key cells in the occurrence of AR. Antisense non-coding RNA in the INK4 locus (ANRIL) was discovered to be involved in the progression of AR. However, the mechanism by which ANRIL mediates the progression of AR remains to be determined. The present study aimed to further explore the mechanism by which ANRIL regulates AR. Thereby, HNECs were treated with IL-13 to mimic AR in vitro. The mRNA expression levels of ANRIL, microRNA (miR)-15a-5p, JAK2, mucin 5AC (MUC5AC), granulocyte-macrophage colony-stimulating factor (GM-CSF) and eotaxin-1, and protein expression levels of JAK2, STAT3 and phosphorylated-STAT3 in HNECs were analyzed using reverse transcription-quantitative PCR and western blotting, respectively. ELISAs were used to detect the secretory levels of inflammatory cytokines and mucin in cell supernatants. In addition, a dual luciferase reporter assay was used to confirm the downstream target of ANRIL and the target gene of miR-15a-5p. The results revealed that the secretory levels of eotaxin-1, GM-CSF and MUC5AC were significantly upregulated by IL-13 in the supernatant of HNECs. The expression levels of ANRIL and JAK2 were also upregulated in IL-13-induced HNECs, while the expression levels of miR-15a-5p were downregulated. In addition, ANRIL was identified to bind to miR-15a-5p. The IL-13-induced upregulation of eotaxin-1, GM-CSF and MUC5AC mRNA expression and secretory levels was significantly inhibited by the genetic knockdown of ANRIL, while the miR-15a-5p inhibitor effectively reversed this effect. JAK2 was also discovered to be directly targeted by miR-15a-5p. The overexpression of JAK2 significantly suppressed the therapeutic effect of miR-15a-5p mimics on IL-13-induced inflammation in vitro. In conclusion, the findings of the present study suggested that the genetic knockdown of ANRIL may suppress the production of inflammatory cytokines and mucin in IL-13-treated HNECs via regulation of the miR-15a-5p/JAK2 axis. Thus, ANRIL may serve as a novel target for AR treatment.

A Drosophila model of neuronal ceroid lipofuscinosis CLN4 reveals a hypermorphic gain of function mechanism.

The autosomal dominant neuronal ceroid lipofuscinoses (NCL) CLN4 is caused by mutations in the synaptic vesicle (SV) protein CSPα. We developed animal models of CLN4 by expressing CLN4 mutant human CSPα (hCSPα) in Drosophila neurons. Similar to patients, CLN4 mutations induced excessive oligomerization of hCSPα and premature lethality in a dose-dependent manner. Instead of being localized to SVs, most CLN4 mutant hCSPα accumulated abnormally, and co-localized with ubiquitinated proteins and the prelysosomal markers HRS and LAMP1. Ultrastructural examination revealed frequent abnormal membrane structures in axons and neuronal somata. The lethality, oligomerization and prelysosomal accumulation induced by CLN4 mutations was attenuated by reducing endogenous wild type (WT) dCSP levels and enhanced by increasing WT levels. Furthermore, reducing the gene dosage of Hsc70 also attenuated CLN4 phenotypes. Taken together, we suggest that CLN4 alleles resemble dominant hypermorphic gain of function mutations that drive excessive oligomerization and impair membrane trafficking.

De novo assembly and characterization of the Hucho taimen transcriptome.

Taimen (Hucho taimen) is an important ecological and economic species that is classified as vulnerable by the IUCN Red List of Threatened Species; however, limited genomic information is available on this species. RNA-Seq is a useful tool for obtaining genetic information and developing genetic markers for nonmodel species in addition to its application in gene expression profiling. In this study, we performed a comprehensive RNA-Seq analysis of taimen. We obtained 157 M clean reads (14.7 Gb) and used them to de novo assemble a high-quality transcriptome with a N50 size of 1,060 bp. In the assembly, 82% of the transcripts were annotated using several databases, and 14,666 of the transcripts contained a full open reading frame. The assembly covered 75% of the transcripts of Atlantic salmon and 57.3% of the protein-coding genes of rainbow trout. To learn about the genome evolution, we performed a systematic comparative analysis across 11 teleosts including eight salmonids and found 313 unique gene families in taimen. Using Atlantic salmon and rainbow trout transcriptomes as the background, we identified 250 positive selection transcripts. The pathway enrichment analysis revealed a unique characteristic of taimen: It possesses more immune-related genes than Atlantic salmon and rainbow trout; moreover, some genes have undergone strong positive selection. We also developed a pipeline for identifying microsatellite marker genotypes in samples and successfully identified 24 polymorphic microsatellite markers for taimen. These data and tools are useful for studying conservation genetics, phylogenetics, evolution among salmonids, and selective breeding for threatened taimen.

[Protective effect of NT-3 gene mediated by hydroxyapatite nanoparticle on the cochlea of guinea pigs injured by excitotoxicity].

OBJECTIVE: To transfect the recombinant plasmid enhancement type green fluorescent protein C2- neurotrophic factor-3 (pEGFPC2-NT3) into the spinal ganglion cells(SGCs) of guinea pigs' cochlea injured by the excitotoxicity of hydroxyapatite particle (HAT), to inject the recombinant plasmid pEGFPC2-NT3 into the guinea pigs' cochlea, and to observe the expression of pEGFPC2-NT3 and the protective effect of pEGFPC2-NT3 on SGCs of the cochlea in guinea pigs. METHODS: The recombinant plasmid pEGFPC2-NT3 with gene-green fluorescent protein was established. Kanic acid (KA) was injected into guinea pigs'cochleae and the excitotoxicity model was established. After a week the recombinant plasmid was transferred into SGCs of guinea pigs'cochlea treated with HAT. The following week the expression of NT-3 was examined by the immunohistochemical method, and the morphology of SGNs was observed under the electronic microscope after 4 weeks, in the mean time the changes of auditory brain-stem response (ABR) were examined. RESULTS: The excitotoxicity models were established successfully. NT-3 expression in the intracytoplasm of SGNs was observed by the immunohistochemical method 1 week after the injection, the morphologic damages of SGNs lessened under the electronic microscope after 4 weeks. ABR was partly restored, compared with ABR after the injury of the excitotoxicity. CONCLUSION: On the 7th day, NT3 gene transferred by HAT through the scala tympani can lessen the excitotoxicity of SGCs after KA was injected into the guinea pigs cochlea.

[Comparison of postoperative drainage and systemic trauma response after endoscopic and traditional near total thyroidectomy].

OBJECTIVE: To investigate the difference of postoperative drainage and systemic trauma response between endoscopic and traditional near total thyroidectomy to provide the basis for selecting appropriate operative methods. METHODS: In this prospective clinical controlled study, 80 patientsscheduled for near total bilateral thyroidectomy for the first time were divided equally into endoscopic surgery group (group A) and open surgery group (group B). The total drainage volume after operation, postoperative extubation time, and postoperative daily drainage volume were recorded after the operation. The contents of triglyceride (TG) and total protein (TP) were determined in the postoperative drainage fluid onthe first day. The levels of interleukin 6 (IL6), high sensitive C reactive protein (HSCRP), alpha 1 acid glycoprotein (AAG), ceruloplasmin (CER) and haptoglobin (HPT) in venous blood were tested before the operation and on the first day after surgery. RESULTS: Compared with those in group B, the postoperative drainage volumein group Aincreased significantly (P=0.000) and the postoperative extubation time was significantly prolonged (P=0.000); the mean postoperative daily drainage volume was significantly larger ingroup A than in group B (P=0.000) and tended to decrease with time in both groups. There was no significant difference in the content of triglycerideortotal protein in the drainage fluid between the two groups on the first day after operation (P=0.429 and 0.324, respectively). In both groups, the contents of AAG, ceruloplasmin and haptoglobin on the first postoperative day were all similar with those measurement before operation (P>0.05), but significant variations occurred in the levels of IL6 and HSCRP on the first postoperative day (P=0.000). The serum levels of IL?6 or HS?CRP did not differ significantly between the two groups on the first day after operation (P=0.054 and 0.066, respectively). CONCLUSION: Compared with open surgery, endoscopic near total bilateral thyroidectomyis associated with an increased the volume of postoperative drainage and a prolonged time of extubationbut not an increased systemic trauma response. Therefore, endoscopic surgery can serve as one of the routine options for patients who are concerned with neckscars resulting from open surgeries.

Hepatic resection versus transarterial chemoembolization for patients with Barcelona Clinic Liver Cancer intermediate stage Child-Pugh A hepatocellular carcinoma.

The present study aimed to compare the overall and recurrence-free survival rates following hepatic resection (HR) and transcatheter arterial chemoembolization (TACE) in patients with Barcelona Clinic Liver Cancer (BCLC) classified intermediate-stage Child-Pugh A hepatocellular carcinoma (HCC). A total of 443 patients were examined, among whom 274 underwent HR, whereas 169 received TACE. The overall survival, recurrence-free survival between groups and subgroups, and risk factors with respect to mortality and recurrence, were analyzed. The 1-, 3- and 5-year overall and recurrence-free survival rates were 70, 46 and 37% and 73, 52, and 37%, respectively after HR, compared with 38, 15, and 12% and 44, 25 and 16%, respectively after TACE. Overall and recurrence-free survival rates were significantly increased following HR compared with TACE. Subgroup analysis in the multi-nodule group showed that the 1-, 3- and 5-year overall survival rates were 68, 38 and 30% after HR, compared with 36, 10 and 0% following TACE. In the solitary tumor group, 1-, 3- and 5-year overall survival rates were 71, 50 and 38% after HR, and 41, 22 and 15% after TACE. The overall survival rate after HR was significantly increased compared with that after TACE in the solitary tumor and multi-nodule groups. The risk factors for mortality include solitary tumor diameter >10 cm, multi-nodules, serum albumin level ≥35 g/l, prothrombin time >13 sec, alphafetoprotein levels >400 ng/ml, and patients with hepatitis B virus. Solitary tumor diameter >10 cm, multi-nodules, and hepatitis B virus (P<0.001) were found to be associated with higher recurrence of HCC. Overall and recurrence-free survival rates were improved after HR compared with those after TACE in BCLC stage B, Child-Pugh A, HCC patients.

[Effects of irrigating solution of Sihuang on morphology and function of nasal mucosa following surgery for chronic sinusitis and nasal polyps].

OBJECTIVE: To investigate the effect of the irrigating solution of Sihuang (ISSH) given after endoscopic surgery for chronic sinusitis and nasal polyps. METHODS: ISSH was used to irrigate the operated sinus cavity for 4 weeks in 109 patients with chronic sinusitis and/or polyps treated by endoscopic surgery, and 0.9%; normal saline (NS) was given in the same manner in another 109 patients receiving ISSH for similar problems, and the effects of the irrigating solutions was compared. Before and 3 months after the surgery, respectively, 32 patients with sinusitis were selected from each group for saccharin test. The mucosa in the ostiomeatal complex region was examined by electron microscopy in 6 patients before, two weeks and one month after surgery, respectively. RESULTS: The total effective rate was not significantly different between the two groups, but the postoperative morphological and functional recovery of the nasal mucosa occurred earlier in ISSH than in NS group (P<0.05). The level of SIgA was similar between the two groups before treatment (P>0.05), whereas after ISSH treatment, the SIgA level was significantly elevated in the treated group. Mucociliary transport rate was similar between ISSH and NS groups before the surgery (3.14+/-1.05 mm/min vs 3.31+/-1.09 mm/min, P>0.05), but 3 month after the surgery, the mucociliary transport rate increased to 6.09+/-2.63 mm/min in ISSH group and 5.04+/-1.22 mm/min in NS group, showing significant difference between them. Under electron microscope, the cilia of the epithelial cells were found exfoliated preoperatively but regularly arranged after the surgery, presenting the "9+2" architecture of the microtubule. CONCLUSION: ISSH used after operation can improve the therapeutic effect of endoscopic surgery for chronic sinusitis and/or nasal polyps.

The complete mitochondrial genome sequence of white-spotted char (Salvelinus leucomaenis).

The complete sequence of the mitochondrial genome of the white-spotted char (Salvelinus leucomaenis) was determined to be 16,658 bp in length, which contains the control region (CR), the origin of light-strand replication (OL), 22 transfer RNA genes, 2 ribosomal genes and 13 protein-coding genes. Overall, base composition of the complete mitochondrial DNA was 28.20% A, 26.51% T, 28.39% C, 16.90% G, with 54.71% AT.

beta-Agkistrodotoxin inhibition of Ca2+-dependent release of glutamate, aspartate, glycine and gamma-aminobutyric acid from cerebrocortical synaptosomes following its binding to synaptic membrane.

The binding properties of beta-AgTX, a snake pre-synaptic toxin, membranes and its effect on transmitter release from cerebrocortical synaptosomes were investigated. Assay of [125I]-beta-AgTX binding to rat synaptic membrane revealed a high affinity binding site for the toxin within the synaptic membrane. Preincubation with beta-AgTX inhibited K+-evoked Ca2+-dependent glutamate release from synaptosomes in a concentration-dependent manner, as determined by an on-line enzyme-linked fluorometric assay. The toxin also blocked the Ca2+-dependent release of other transmitters, aspartate, glycine, and GABA induced by K+-depolarization. However, Ca2+-ionophore, ionomycin-stimulated Ca2+-dependent transmitter release was not significantly affected by beta-AgTX, indicating that the toxin inhibits transmitter release by reducing the entry of Ca2+ into cytoplasm. It is suggested that beta-AgTX-binding site in synaptic membrane is related to the release of a variety of transmitters.

[Excitotoxic effect of glutamate on the afferent neurons in guinea pigs].

OBJECTIVE: To compare Glu-induced excitotoxic damage to the cochlear afferent neurons and changes of cochlear electrophysiology by directly injecting two different concentrations of glutamate into the inner ear of adult guinea pigs. METHODS: Guinea pigs were injected high doses glutamate (Glu-H,20 mmol/L, 10 microl) and low doses of glutamate (Glu-L, 10 mmol/L, 10 microl) into the perilymph of scala tympani. The compound action potential (CAP) was recorded and pathologic changes were evaluated with light and transmission electronic microscope at various time points after the application of glutamate. Results The thresholds of the CAP of Glu-L group (10 mmoL/L, less than one day) and Glu-H group (20 mmol/ L) were significantly increased compared with those of the normal group (P <0.001). But the thresholds of CAP of glutamate application (10 mmol/L, more than one week) weren't significantly different from those of the normal group (P > 0.05). Some changes of the spiral ganglion cells of the cochlear axis were observed in the 2 drug groups by light microscope. There were some vacuoles in the cell body and condensation of nuclear chromatin, shallow cellar liquid. The ratios of pathological changes between the control group and Glu-H group of different concentrations were notably different (P < 0.05). The observation under transmission electronic microscope found disvolution or necrosis of afferent nerve endings, myelinated nerve fibers, and model spiral ganglion cells. The pathological changes of the Glu-H group were more serious than those of the Glu-L group (10 mmol/L). The pathological changes of the Glu-L group would recover in 4 weeks, but those of the drug group would gradually deteriorate and degeneration would become necrosis. CONCLUSION: The morphologic changes and functional losses of afferent neurons in guinea pigs were caused by glutamate in scala tympani. The low dose (10 mmol/L) had induced reversible damage to cochlear afferent neurons. The high dose (20 mmol/L) had triggered irreversible neurodegeneration.

Appropriate treatment strategies improve survival of hepatocellular carcinoma patients with portal vein tumor thrombus.

AIM: To evaluate the survival benefits of different treatment strategies for hepatocellular carcinoma (HCC) patients with portal vein tumor thrombus (PVTT) and to determine the prognosis factors. METHODS: Between 2007 and 2009, 338 HCC patients treated for PVTT were retrospectively studied. The patients were divided into 4 groups that underwent different treatments: the conservative treatment group (n = 75), the transarterial chemoembolization (TACE) group (n = 86), the hepatic resection group (n = 90), and the hepatic resection associated with postoperative TACE group (n = 87). Survival rates were determined using the Kaplan-Meier method and differences between the groups were identified through log-rank analysis. Cox's proportional hazard model was used to identify the risk factors for survival. RESULTS: The mean survival periods for patients in the conservative treatment, TACE, hepatic resection and hepatic resection associated with postoperative TACE groups were 3.8, 7, 8.2 and 15.1 mo, respectively. Significant differences were observed in the survival rates. For the surgical resection associated with postoperative TACE group, the survival rates after 1, 2 and 3 years were 49%, 37% and 19%, respectively. These results were significantly higher than those of the other groups (P < 0.05). Meanwhile, the 1, 2 and 3 year survival rates for the surgical resection group were 28%, 20% and 15%, whereas those for the TACE group were 17.5%, 0% and 0%, respectively. These values significantly increased after hepatic resection compared with those after TACE (P < 0.05). CONCLUSION: Surgical resection is the most effective therapeutic strategy for HCC patients with PVTT and results in high hepatic functional reserve. For patients who can tolerate the procedure, postoperative TACE is necessary to prevent recurrence and prolong the survival period.

A polycaprolactone-tricalcium phosphate composite scaffold as an autograft-free spinal fusion cage in a sheep model.

Titanium (Ti) based spinal fusion cages are frequently used in the clinics for the treatment of spinal degeneration and related diseases, however, their further clinical application is generally harassed by several drawbacks such as stress shielding, non-biodegradability and additional bone grafting procedure. Our earlier work has demonstrated the efficacy of a biodegradable macro-porous polycaprolactone-tricalcium phosphate (PCL-TCP) composite scaffold in promoting bony tissue ingrowth as well as its ability to sustain mechanical loads upon implantation into an orthotopic defect site. In this study, we investigated the use of PCL-TCP scaffold as an autograft-free spinal fusion cage in a preclinical sheep model over 12 months, and compared the fusion efficacy against Ti cages incorporated with autografts. Results showed that despite PCL-TCP scaffold as an autograft-free cage attaining a slower fusion rate at early stage (6 month), it achieved similar degree of spinal fusion efficacy as Ti cages aided with autograft at 12 month post-operation as evidenced by the radiographic and histological evaluation. PCL-TCP cages alone demonstrated better bone ingrowth with 2.6 fold higher bone/interspace ratio (B/I) and more homogeneous bone tissue distribution compared with that of the Ti cages (88.10  ±  3.63% vs. 33.74  ±  2.78%, p < 0.05) as seen from the histological and micro-CT analysis. Moreover, besides the bone tissue ingrowth, a quantitative approach was illustrated to accurately evaluate the osteointegration of fusion cage with surrounding bone tissue, and showed a 1.36 fold higher degree of osteointegration occurred in PCL-TCP cage group than Ti cage group (CS/PC: 79.31  ±  3.15% vs 58.44  ±  2.43%, p < 0.05). Furthermore, biomechanical analysis showed comparable mechanical strength of fused segments in both groups in terms of the range of motion and stiffness at 12 month (p > 0.05). The degradation profile of the PCL-TCP cages was noted to increase in tandem with new bone ingrowth into the pores, while maintaining good structural integrity necessary for supporting the spinal interbody segments. Therefore, with the better osteointegration, more bone tissue ingrowth as well as its favorable biodegradable and radiolucent properties, PCL-TCP cage has been demonstrated to be a promising candidate as an autograft-free fusion cage for clinical application.

[Optimal feeding strategy for juvenile Hucho taimen].

Three experiments including starvation and re-feeding, starvation and re-feeding recovery, and feeding frequency per day were conducted to approach the optimal feeding strategy for the growth and survival of juvenile Hucho taimen. In the experiment of starvation and re-feeding, all groups of restricted feeding showed non-compensatory growth. However, in the experiment of starvation and re-feeding recovery, different degrees of compensatory growth appeared in different starving groups, among which, the half a day starvation and half a day feeding group (S1/2) had a weight increment approximately the same as the control, and showed completely compensatory growth, indicating that the S1/2 could be a useful feeding strategy for the juvenile H. taimen at its early growth stage with the body mass from 0 to 2 g and at the water temperature from 9 to 15.3 degrees C. In feeding frequency experiment, the group T3 (three meals per day) had the highest body length, body mass, specific growth rate, and relatively high food conversion ratio, indicating that three meals a day could be more effective for improving the growth performance of juvenile H. taimen at its late stage with the body mass from 2 to 21 g and at the water temperature from 8.8 to 15.5 degrees C.

[Effects of delayed first feeding on growth and survival of Hucho taimen larvae].

This paper studied the effects of delaying first breeding Hucho taimen larvae for different days on the larvae growth, survival, and body size. Five treatments were installed, i. e. , feeding begins on the first eating day (control, S0) and on the 9th, 12th, 15th, and 18th days after the first eating day (S1 -S4) at 10.4-14.9 degree C, respectively. By the end of the experiment (36-day), the growth rate and initial feeding rate in S1 was higher than that in S0, and the overall mortality rate in S1 was lower, but the body size and mass in S1and S0 had no significant difference. Compared with S0, S2 had higher growth rate, initial feeding rate, total mortality, and self-mutilation mortality, the body mass was significantly lower, but the body size had less difference. S3 had higher first feeding rate, body size, total mortality, and self-mutilation mortality, but significantly lower body mass than S0, whereas the growth rate had less difference. In S4, the growth rate and body mass were lower, and the total mortality and self-mutilation mortality were higher than those in S0. It was suggested that under the same conditions, delaying first feeding for 9 days would induce H. taimen larvae presenting "completely compensatory growth", and this feeding way could be applied for the culture of H. taimen larvae in their initial feeding period.

The treatment effect of porous titanium alloy rod on the early stage talar osteonecrosis of sheep.

Osteonecrosis of the talus (ONT) may severely affect the function of the ankle joint. Most orthopedists believe that ONT should be treated at an early stage, but a concise and effective surgical treatment is lacking. In this study, porous titanium alloy rods were prepared and implanted into the tali of sheep with early-stage ONT (IM group). The curative effect of the rods was compared to treatment by core decompression (DC group). No significant differences in bone reconstruction were observed between the two groups at 1 month after intervention. After 3 months, the macroscopic view of gross specimens of the IM group showed ordinary contours, but the specimens of the DC group showed obvious partial bone defects and cartilage degeneration. Quantitative analysis of the reconstructed trabeculae by micro-CT and histological study suggested that the curative effect of the IM group was superior to that of the DC group at 3 months after intervention. These favorable short-term results of the implantation of porous titanium alloy rods into the tali of sheep with early-stage ONT may provide insight into an innovative surgical treatment for ONT.