Objective:Through histological analysis, immunofluorescence staining, electrophysiological detection and Sensory and motor function evaluation to investigate the effects of 3D printed hydrogel scaffold combined with bone marrow mesenchymal stem cells (BMSCs) in promoting functional recovery of spinal cord injury.Methods:10% GelMA hydrogel and 10 6 U stem cell suspension were prepared into bioink of appropriate concentration to construct the biomimetic spinal cord scaffold through 3D printing platform. The scaffold was placed in the medium and cultured in an environment of 37 ℃ CO 2 incubator. The microstructure of the scaffolds and the distribution of BMSC in the scaffolds was observed by scanning electron microscope. CAM/PI staining and confocal microscopy were used to observe the survival of stem cells in the scaffolds and determine the biocompatibility of the scaffolds. The scaffolds were implanted into the subcutaneous tissues of the back of rats, and the subcutaneous tissues were determined by HE staining to detect the immunogenicity of the scaffolds. After the rat model of hemicytoma defect was made, stents were transplanted for treatment, and confocal microscopy was used to evaluate the regeneration of neurons and axons in local area of spinal cord injury. At the same time, BBB score was used to evaluate motor function, mechanical pain score was used to evaluate sensory function, and surface electrode detection method was used to evaluate electrophysiological recovery weekly. Results:The long spindle shaped BSMC were uniformly distributed in the scaffold with a loose reticular structure. The scaffolds had good biocompatibility, and the cell survival rate of the prepared scaffolds reached 96% after 24 hours of printing. After 28 days of subcutaneous transplantation, the immune rejection was mild and immunogenicity was low. It was shown that the regenerated spinal cord tissue in the treatment group was significantly increased compared with the control group, which was widely distributed with cells after 28 days by HE staining. It was confirmed that part of the regenerated spinal cord tissue was neurons by immunohistochemical staining.Compared with the injured group, the regeneration of neurons and axons in the treatment group were significantly increased by immunofluorescence staining and confocal microscopy. In the treatment group, the BBB score recovered to 10 points, while the control group only recovered to about 1 point in the first week, which was statistically significant. And it recovered to 17 in the fourth week, while the control group only recovered to about 4 point in the four week, which was statistically significant. The Angle of inclined plate support of the treatment group was restored to 40 degrees, while it was only restored to 22 degrees in the control group. The pain threshold of the treatment group decreased to 18.5 points, which was not statistically different from that of the control group. The latent recovery effect of electrophysiology in the treatment group was the same as that in the sham operation group and better than that in the control group.Conclusion:3D printing hydrogel scaffold with loose network structure is suitable for cell proliferation. It has well biological survival, low cytotoxicity and low immunogenicity, which promoted neurons and axons to recovery and extend so as to effectively promote the recovery of motor function, sensory function and neural signal transmission rate after spinal cord injury.

BACKGROUND: Emerging studies have focused on cell transplantation. Schwann cells (SCs) can secrete various neurotrophic factors and improve local environment around injury. Plenty of documents have demonstrated that SCs could promote functional recovery following spinal injury. Many transplanting methods are available for treating spinal cord injury, and the intravenous cell transplantation is profitable for easy operation and avoidance of additional trauma. OBJECTIVE: To investigate the effects of intravenous transplantation of SCs on spinal cord injury in rats. METHODS: The bilateral sciatic nerves of Wistar rats were separated in vitro, cultured by tissue clot method, identified by S-100 and labeled by Hoechst33342. Sixty rat models with T10 spinal cord injury were prepared using impactor model- II type weight drop apparatus. Then the injured rats were randomly divided into 3 groups: blank control, DMEM control and SCs transplantation groups. No treatment was performed in the blank control group. Totally 1 mL DMEM and or SCs was injected into rats of DMEM control and SCs transplantation groups by tail vein respectively. Basso Beattie Bresnahan (B6B) scores were performed at 1 day before and 1, 3 days, 1 week and weekly after operation. The migration of transplanted SCs was observed at 2 weeks and 4 after transplantation. The expressions of glial fibrillary acidic protein (GFAP) and neuron specific enolase (NSE) were detected by haematoxylin-eosin staining and immune-fluorescence staining.RESULTS AND CONCLUSION: The purity of SCs reached 95%. Hoechst33342 positive cells were observed throughout the injured and the nearby region of spinal cord at 1, 2, and 4 weeks after transplantation. The statistical difference of BBB score among the SCs transplantation, blank control, and the DMEM control groups displayed at 4 weeks after transplantation (P < 0.05), and the BBB scores of the SCs transplantation were higher than other groups. Haematoxylin-eosin staining showed the cavity formed in each group at 8 weeks after transplantation, but the area of SCs transplantation was smaller than that of the blank control and DMEM control groups. The immunofluorescence staining indicated that the expression of GFAP were more intense in the blank control group and DMEM control than SCs transplantation (P < 0.05), while the expression of NSE was more intense in SCs transplantation than other groups (P< 0.05). It implied that intravenous transplantation of SCs promotes regeneration of axon and improves neurological functions after spinal cord injury in rats.

Objective To investigate the surgical techniques and clinical efficacy of cannulated screws combined with transposition of cuboid periosteal flap pedicled with fascia and lateral tarsal artery in the treatment of talus neck fractures.Methods From March 2008 to June 2011,12 cases with talus neck fractures were treated with cannulated screws combined with transposition of cuboid periosteal flap pedicled with fascia and lateral tarsal artery.External fixation in functional position for 12-14 weeks,and the load time was determined by X-ray fracture healing.Functional results were assessed according to AOFAS (American Orthopaedic Foot and Ankle Society) score at last follow-up.Results Twelve cases were followed up for an average of 24 months (12 to 48 months).One case of skin flap necrosis healed by dressing.All the fractures healed,and the mean healing time was 20 weeks (16 to 24 weeks).AOFAS ankle and hindfoot mean postoperative score was 82.5 (ranging from 55 to 96) points.Among them 4 cases were excellent,good in 5 cases,fair in 3 cases.Two cases presented with mild posttraumatic subtalar arthritis and pain relief after oral anti-inflammatory analgesic.One case with avascular necrosis of talus body,and X-ray displayed sclerosis of talus body,but no collapse,and the patient was told to reduce weight and regularly followed up.Conclusion The application of cannulated screws combined with transposition of cuboid periosteal flap pedicled with fascia and lateral tarsal artery for the treatment of talus neck fractures is an effective treatment,which can improves the blood supply of the talus body and reduces the occurrence of avascular necrosis of the talus.

To our knowledge, there has been no clinical report of artesunate in the treatment of lung cancer. This study was designed to compare the efficacy and toxicity of artesunate combined with NP (a chemotherapy regimen of vinorelbine and cisplatin) and NP alone in the treatment of advanced non-small cell lung cancer (NSCLC).

ObjectiveTo investigate the characteristics of traumatic spinal cord injury (TSCI) urban inpatients of Tianjin in 2007. MethodsInpatients with TSCI of 8 hospitals in Tianjin in 2007 were reviewed. ResultsThere were 73 patients in total. Mean age was (51.34±14.597) years. Male∶Female was 3.56∶1. Falling, motor vehicle accidents （MVC） were the main causes of TSCI. The cervical spinal cord injuries were predominant. 26% were complete injury and 74% were incomplete. 6 cases were dead. Patients with ASIA grade D recover well. ConclusionFor the TSCI, the ages of patients increases and falling is the main cause.

The aim of this study is to establish a simple and accurate method for vaccine immune e-valuation of porcine pseudorabies virus.In this research,PRV-gD recombinant protein was ex-pressed from mammalian cell HEK-293F as coating antigen,and then the reaction conditions of gD-iELISA were optimized according to checkerboard titration method.The gD-iELISA was used to detect the antibody levels of 211 clinical pig serum samples and the consistency with the neu-tralizing antibody levels wasanalyzed.The results showed that the antigen coating concentration was 0.90 mg/L;the serum to be detected was diluted 1∶100 and incubated at 37 ℃ for 30 min;goat anti-pig IgG-HRP antibody was diluted 1∶55 000 and incubated at 37 ℃ for 30 min;TMB sub-strate was developed at 37 ℃ for 20 min.The method could detect 1∶6 400 diluted PRV positive serum.The results of CSFV,PRRSV,PCV-2,PEDV and FMDV positive sera were all negative by gD-iELISA,and there was no cross-reaction between the method and the above positive sera.The coincidence rate of gD-iELISA and commercial kits was 95.26％,and the intra-and inter-batch co-efficients of variation were both less than 10％.Correlation analysis showed that the correlation coefficient(r)between gD antibody level and neutralizing antibody level was significantly greater than that of gB antibody level,and the gD antibody level had a good linear relationship with the neutralizing antibody level.The results indicated that gD-iELISA was more suitable for vaccine im-mune evaluation of PRV than gB-iELISA.Therefore,the method will have a good prospect of ap-plication in the immunization control of the PRV.

Objective To detect the change of the anti-S. japonicum antibody level after people migrated from outside embankment to newly established town. Methods Three pilot spots were established for the investigation: one spot thut both inhabitancy and cultivation disused (A), one spot that only inhabitancy disused but farming continued (B) and the third one served as control (C). DIGFA and ELISA were used to detect the antibody level in the populations from 2002 to 2005. Results The positive rate of anti-S.japonkum antibody declined significantly from 6.63% to 3.52% by DIGFA and from 7.26% to 3.71% by ELISA at spot A (X2=5.2625, P

OBJECTIVETo discuss radiological characteristics and clinical manifestation of isolated lumbar foraminal stenosis.

METHODSFrom March 2011 to March 2014, 21 patients with isolated degenerative lumbar foraminal stenosis accepted lumbar decompression and fusion in Beijing Luhe Hospital. Intervertebral disc space was evaluated by measuring the position of joint-body line on preoperative X-ray. Bilateral foraminal area of the corresponding segment in CT (sagittal view of 2D reconstruction) and MRI (T2W1 sagittal view) were measured by Surgimap software. For patients with unilateral symptoms, foraminal area of the affected side was compared with that of the contralateral side. Foraminal area of the same segment on CT was also compared with that on MRI. Preoperatively and at the final follow-up, visual analogue score (VAS) and Oswestry Disability Index (ODI) were used to evaluate clinical outcomes.

RESULTSAll patients had a follow-up over 6 months and the average follow-up was 16.8 months (7-42 months). Of the 21 patients (26 segments), 12 segments showed gross narrowing and 14 segments showed slight narrowing. After preoperative measurement on MRI, 6 patients had foraminal stenosis of grade 2, and 15 patients had foraminal stenosis of grade 3, showing no significant difference in clinical outcomes. Compared with the foraminal area of the unaffected side, the affected side showed a decrease of 16% on CT and 28% on MRI, and the difference was statistically significant (t = 3.453, P < 0.05). The foraminal area measured on CT was larger than that measured on MRI (P < 0.05). Compared with that preoperatively, VAS (back pain), VAS (leg pain) and ODI showed significant improvement at the final follow-up (P < 0.05).

CONCLUSIONSRadiological examinations as X-ray, CT, MRI and intervertebral foramen block technique play an important role in the diagnosis of foraminal stenosis. Soft oppression caused by hyperplasia and hypertrophy of transforaminal ligment or joint capsule may be important promoters of degenerative lumbar foraminal stenosis. Lumbar foraminal decompression and interbody fusion can satisfactorily improve preoperative symptoms.

Constriction, Pathologic ; diagnostic imaging ; Decompression, Surgical ; Humans ; Lumbar Vertebrae ; surgery ; Lumbosacral Region ; Magnetic Resonance Imaging ; Spinal Fusion ; Spinal Stenosis ; diagnostic imaging ; Tomography, X-Ray Computed

The aim of this study was to investigate the correlation between porcine epidemic diar-rhea virus(PEDV)S1 IgG antibody levels and neutralizing antibody potency in sow sera.Sera from 5 PEDV-infected farms with a clear immune background,5 non-infected farms and 5 infected farms with an unclear immune background,and sera from return-fed reserve pigs,totaling 716 copies,were collected and measured,and the correlation between PEDV S1 IgG antibodies and neutralizing antibodies was analyzed.The results showed that the PEDV S1 IgG and neutralizing antibodies of sow sera showed highly significant positive correlation,the correlation coefficient was 0.892(P＜0.000 1).Previous studies have shown that the level of PEDV neutralizing antibodies in sow serum correlates with the ability of piglets'maternal antibodies to resist PEDV infection.Therefore,the a-bility of maternal antibodies against PEDV in piglets can be evaluated by detecting PEDV S1 IgG antibodies in the serum of sows.In 10 PEDV-infected farms,the neutralizing antibodies to PEDV in the sera of sows after immunization were generally high,and the S1 IgG antibodies were also high,and their S/P values were higher than 3.5 in 66.9％of the farms(347/519),and the highest anti-body levels were found in the four farms in which PED did not occur,whereas the neutralizing an-tibodies in the immunized sows in the five PEDV-uninfected farms were generally low,and their S1 IgG antibodies were also low,and only 8.1％(13/161)having S/P values higher than 3.5.The re-sults suggest that most sows in PEDV-infected farms can provide good immunoprotection to pig-lets after immunization,while pigs in PEDV-uninfected farms need further immunization if they need to achieve a higher level of immunoprotection.The present study provides a substantial clini-cal basis for the use of PEDV S1 IgG antibody levels to assess the effectiveness of PEDV antibody protection in swine herds.

Objective To establish a stable rat model of non-obese polycystic ovary syndrome(PCOS)with clinical characteristics.Methods Dehydroepiandrosterone(DHEA)was used to establish a PCOS rat model by subcutaneous injection.Three-week-old female SD rats were divided into a normal group,6 mg/kg DHEA model group,and 60 mg/kg DHEA model group.The model groups were subcutaneously injected with the corresponding dose of DHEA daily,while the normal group was subcutaneously injected with glycerol daily for 21 consecutive days.The model was evaluated with ovarian histopathology as the gold standard to determine the optimal dosage of DHEA to induce a PCOS rat model.On this basis,the optimal DHEA modeling dose was selected,and stop and continue modeling groups were set up to observe the model for 28 days and evaluate its maintenance.The stop modeling group was no longer given DHEA,and the continued modeling group was subcutaneously injected with 60 mg/kg DHEA every 48 h.The evaluation indicators included body mass,estrous cycle,fasting blood glucose,serum insulin,histopathologic morphology of the ovaries,and serum sex hormone levels.Results(1)Compared with the normal group,the 6 mg/kg and 60 mg/kg DHEA model groups showed no significant difference in body mass,and their estrous cycles were irregular.There were more cystically dilated large follicles in the ovaries;fewer mature follicles;reduced layers of granulosa cells,which were arranged in a sparse and disorganized manner;and fewer lutea in the 6 mg/kg and 60 mg/kg DHEA model groups than the normal group.Furthermore,serum T and E2 levels were significantly higher in the 60 mg/kg DHEA model group(P＜0.05)than the normal group.(2)The stop modeling group(A2 group)resumed regular estrous cycles after 2 weeks,various growth follicles and corpora lutea were observed in the ovarian tissues,the number of cystic follicles was reduced,the number of granulosa cell layers increased,mature follicles were visible,oocyte morphology was locally intact,and the levels of E2 and AMH were reduced compared with the normal group(A1 group)(P＜0.05).(3)The continue model group(B2 group)was in the late stage of estrous cycle for a long period,and there were more large follicles with cystic dilatation,fewer mature follicles,fewer layers of granulosa cells with a sparse and disordered arrangement,and significantly fewer corpus lutea in the ovaries compared with the normal group(B1 group).The levels of serum LH,LH/FSH,and T were elevated(P＜0.05).Conclusions Subcutaneous injection of 60 mg/kg DHEA for 21 consecutive days can be used to successfully construct a non-obese PCOS rat model that possesses clinical characteristics.Subcutaneous injection of 60 mg/kg DHEA every 48 hours maintains the stability of the model.