ObjectiveTo obtain a construct chimeric antigen receptor macrophages （CAR-M） targeting disialoganglioside GD2 （GD2） and harboring the intracellular domain of macrophage-specific activating signaling adaptor protein DNAX-activating protein 12 （DAP12）. MethodsA CAR molecule targeting GD2 with the DAP12 intracellular domain as the signaling module ［Anti-GD2 CAR （DAP12）］ was designed， synthesized， and cloned into an adenoviral vector. This virus was used to infect PMA-differentiated THP-1 macrophages to construct green fluorescent protein （GFP）-labeled Anti-GD2 CAR THP-1 cells. CAR expression was confirmed using live-cell imaging. In vitro co-culture assays were performed to evaluate the phagocytic function of empty， Anti-GD2 CAR-M （CD3ζ）， and Anti-GD2 CAR-M （DAP12） groups against glioma cells with high （T98G） and low （U251） GD2 expression. ResultsThe CAR adenovirus efficiently transduced THP-1 macrophages， resulting in successful generation of Anti-GD2 CAR-M. Flow cytometry analysis following co-culture with glioma cells showed that compared to the empty group， the Anti-GD2 CAR-M （CD3ζ） group exhibited significantly enhanced phagocytosis （P<0.05）. Furthermore， compared to the Anti-GD2 CAR-M （CD3ζ） group， the Anti-GD2 CAR-M （DAP12） group demonstrated a further increase in phagocytic capacity specifically against GD2-high T98G cells （P<0.05）. ConclusionAnti-GD2 CAR-M （DAP12） cells are successfully established by transducing PMA-differentiated THP-1-M with recombinant Anti-GD2 CAR （DAP12） adenovirus. The engineered CAR-M can specifically target phagocytose glioma cells with high GD2 expression.

ObjectiveTo obtain stably transfected chimeric antigen receptor macrophages （CAR-M） targeting disialosylganglioside （GD2） based on the constitutive expression of myeloid differentiation factor 88 （MYD88）. MethodsAn Anti-GD2 CAR adenoviral vector incorporating the CD3ζ （cluster of differentiation 3 zeta chain）-MYD88 fusion construct was designed and constructed. THP-1-derived macrophages （THP-1M）， induced by phorbol 12-myristate 13-acetate （PMA） differentiation， were infected with this adenovirus to generate Anti-GD2 CAR THP-1 cells bearing a green fluorescent protein （GFP） tag. CAR expression was monitored using a live-cell imaging system. The phagocytic function of Empty CAR-M， Anti-GD2 CAR-M（CD3ζ）， and Anti-GD2 CAR-M（CD3ζ-MYD88） groups against glioma cells was evaluated in vitro through co-culture experiments， and their polarization status was also analyzed. ResultsCAR adenovirus successfully transfected THP-1 to obtain Anti-GD2 CAR-M. After co-culture with glioma cells， flow cytometry results showed that compared with the Empty CAR-M group and the Anti-GD2 CAR-M（CD3ζ） group， the Anti-GD2 CAR-M（CD3ζ-MYD88） group exhibited enhanced phagocytosis against T98G cells with high GD2 expression （P<0.05） and also showed upregulation of CD86 （P<0.05）. Compared with the Empty CAR-M group and the Anti-GD2 CAR-M（CD3ζ） group， the Anti-GD2 CAR-M（CD3ζ-MYD88）CAR-M group exhibited enhanced phagocytosis against T98G cells with high GD2 expression （P<0.05） and also showed upregulation of CD86 （P<0.05）， and the expression of inflammatory cytokines interleukin （IL）-6 and IL-12 was increased in this group （P<0.05）. ConclusionThis study successfully constructed Anti-GD2 CAR-M cells and verified their phagocytic activity， demonstrating that constitutive expression of MYD88 can promote inflammatory polarization of CAR-M. This provides a potential structural modification approach for the development of pro-inflammatory CAR-M.

Objective: To construct microRNA(miR)-22 gene knockout(miR-22-/-) mice using CRISPR/Cas 9 technology, to breed miR-22-/- mice and to identify their genotypes. Methods : In this experiment, CRISPR/Cas 9 technology was used to construct miR-22-/- genetically engineered mice. After gene identification, the F0 generation miR-22-/- mice were mated with wild-type mice in the same litter to obtain F1 generation miR-22-/- mice. The miR-22 knockout efficiency was analyzed at the RNA level by real-time fluorescence quantitative polymerase chain reaction(qPCR). Western blot was used to detect the interaction between miR-22 and target genes. Results : miR-22-/- mice were successfully constructed using CRISPR/Cas 9 technology, gene identification was performed on the bred mice, and three stable genotypes of miR-22+/+,miR-22+/-, and miR-22-/- were identified. The real-time fluorescence quantitative PCR detection results confirmed that miR-22-/- mice showed almost no expression of miR-22 in the heart, liver, lung, kidney, spleen, and thymus tissues compared to wild-type mice in the same litter. Western blot analysis showed that the relative expression level of NLRP3 protein in miR-22-/- mouse tissues was lower than that in wild-type mice. Conclusion A miR-22-/- mouse model is successfully constructed, and stable genetic homozygous miR-22-/- mice is obtained. This indicates that miR-22 has an inhibitory effect on the downstream target gene NLRP3.

Notoginseng Radix et Rhizoma as a traditional Chinese herbal medicine has now been recognized and paid attention to by the pharmaceutical community.Modern phytochemical studies have shown that Panax notoginseng saponin is the main chemical compo-nent of Notoginseng Radix et Rhizoma.Modern pharmacological studies and clinical applications have revealed that it has anti-cancer,antioxidant and cardiovascular disease effects.In this study,we reviewed the research progress of the main chemical components and pharmacological effects of Notoginseng Radix et Rhizoma,with the aim of providing assistance for the clinical application and later stud-ies of Notoginseng Radix et Rhizoma.

Objective:To explore the different design of readiness assurance process (RAP) and application in TBL teaching of rehabilitation medicine, and to compare the teaching effects.Methods:A total of 40 students who had clinical rotation in our department were chose as research subjects, and they were taught with TBL teaching. Two chapters of "Stroke Rehabilitation" and "Spinal Cord Injury Rehabilitation" were chosen for the two lectures. During the course of "Stroke Rehabilitation", the RAP adopted open-ended questions, and the application part simulated Teamwork meetings. In the course of "Spinal Cord Injury Rehabilitation", the RAP used close-ended multiple-choice questions, and the application part simulated making rehabilitation plan. At the end of the two lectures, questionnaires were send to students.Results:For the RAP part, more students preferred open-ended questions. In the application part, students preferred to make rehabilitation plan.Conclusion:The process of clinical diagnosis and treatment and the formulation of rehabilitation plan in clinical practice of rehabilitation medicine has been applied to different parts of TBL teachings, and students are favor of this teaching method, which provides a reference for the future TBL teaching design of rehabilitation medicine.

Objective To compare the curative effect of fixation of adolescent tibial intercondylar emi-nence fracture among suture anchor,hollow screw and wire. Methods Forty-six adolescent cases of the tibial intercondylar eminence fracture treated with surgical treatment were selected from January 2010 to June 2016 and divided into three groups refer to intra-operative fixation suture anchor group(Group A),hollow screw group (Group B)and wire group(Group C).Duration of treatment,total operation time,hospital stay and surgery times were recorded. All patient condition was assessed with the Lysholm,Tegner,IKDC and VAS score. Results All patients received an average of 13(11~14)months follow-up visit. No blood-vessel,nerve and osteoepiphysis injured,infection and fracture displacement occurred.Before receiving treatment,difference in Lysholm,Tegner, IKDC and VAS score of group A,B and C showed no statistical difference.When it comes to hospitalization condi-tion,data were as follows.Group A/B/C:operation time(80.67 ± 16.68/114.00 ± 20.28/111.88 ± 20.07)min, hospital stay(8.40 ± 1.12/ 15.47 ± 1.25/ 15.19 ± 1.17)d,surgery times(1/2/2)times. Moreover,compared with those before operation and after operation in both of groups,the Lysholm,Tegner,IKDC and VAS score were improved(P < 0.05). Besides,the Lysholm,Tegner,IKDC and VAS score of group A,B and C did not have statistically significant difference yet after post treatment(P>0.05).Conclusions The curative effect of fixation of adolescent tibial intercondylar eminence fracture among suture anchor,hollow screw and wire was similar. By contrast,the fixation of fracture by use of suture anchor can decrease operation time and hospital stay to some extent. It had advantage of need not to have a second operation to remove the internal fixation and can be used in preference.

Objective To investigate gender differences in the curative effects of allograft and autologous hamstring tendon in anterior cruciate ligament reconstruction. Methods One hundred and forty-eight patients with the anterior cruciate ligament injury received surgical treatment were enrolled from January 2011 to January 2015 and divided into two groups,including group M(Men)and group W(Women). Patients in each group were also divided in the allograft group (Group A) and the autologous hamstring tendon group (Group B). The patient condition was separately assessed with the Lysholm,Tegner,VAS score and the value of KT-1000 before and after surgery. Results All patients received an average of 16(13-18)-month follow-up visit. No severe complication , such as blood-vessel and nerve injury ,infection and rupture of graft ,occurred in patients after operations. Compared with those before operation,the Lysholm,Tegner,VAS score and the value of KT-1000 after operation in four groups were significantly improved (P < 0.05,respectively). However,no significant differences were found in the Lysholm,Tegner,VAS score and the value of KT-1000 improvement among group MA,MB,WA and WB post treatment(P>0.05). Conclusions The curative effect of ACL reconstruction between allograft ligament and autologous hamstring tendon has no significant difference in gender. The selection of graft should be considered in more aspects.

Objective To compare and analyze the curative effects of mediopatellar plica syndrome under different therapies. Methods 147 patients with mediopatellar plica syndrome were enrolled in the study from January 2011 to January 2015 and divided into three groups refer to treatment: Group A, B and C. Each group was also divided in the youth group and the mid-age group. Different treatments were used in the three groups. Group A: drug therapy and physicotherapeutics; Group B: intraarticular injection regularly; Group C: arthroscopic surgical treatment. All patient conditions were assessed with Lysholm score. Results All patients received an average of 11 (9 ~ 13) months follow-up. No blood-vessel and nerve injured and infection occurred. Before receiving treatment, difference in Lysholm score of the three groups showed no statistical difference (F = 0.08, P = 0.926); after treatment, difference revealed significant difference (F = 15.48, P = 0.001). Compared with those before operation and after operation among the three groups, the Lysholm score was improved (tA = 3.43, tB = 6.74, tC = 7.99, P = 0.001). In the rangeability of Lysholm score, the Group C > B > A (F = 66.43, tAB = 5.97, tAC = 11.52, tBC = 5.55, P = 0.001);The general youth group > the general mid-age (t = 7.91, P = 0.001). Conclusion The best therapeutic method for mediopatellar plica syndrome is the arthroscopy. As for mid-age patients, it is necessary to inform them of possibility that prognosis is not well in preoperative planning.

Objective To investigate bone mineral density ( BMD ) after filling in the bone scaffolds with anti-tuberculosis controlled-release microspheres, and provide experimental basis for decrease of the side effects of anti-tuberculosis therapy after spinal surgery.Methods The bone densitometer was used to observe the changes of bone mineral density before and after the infusion with the artificial allograft bone (Group A), the controlled release complex of the RFP controlled-release microspheres-artificial allograft bone (Group B), and RFP-artificial allograft bone complex (Group C), respectively.Results BMDs of three groups before perfusion were not different significantly [Group A:(0.191 ±0.018)g/cm2;Group B:(0.186 ±0.016)g/cm2;Group C:(0.189 ±0.018)g/cm2;P >0.05].BMDs of three groups after perfusion were not different significantly [Group A:(0.191 ±0.018)g/cm2;Group B:(0.179 ±0.023)g/cm2;Group C:(0.185 ±0.021)g/cm2;P >0.05].Conclusions RFP microspheres using ultrasonic vibration method and the porous bone were prepared to controlled-release anti-tuberculosis complex .BMD of three groups after perfusion were not influenced obviously .

Objective To investigate the clinical value of 18 F-FDG PET CT scan in the diagnosis of thin lung nodules. Methods 56 solitary cases of nodules in the lungs,nodule size ranged from 1～3 cm,conducted 18 F-FDG PET/CT imaging and thin layer CT scan at the same time, 18 F-FDG PET imaging using visual measurement combined with semi-quantitative method to determine the degree of malignant of the lesions; conducted MPR recon-struction after thin layer CT scan, based on the lesions morphology to determine the degree of malignant. Then com-pared the results of above,and the results of the combination of 18 F-FDG PET imaging and thin layer CT scan,with the pathological results after surgery. Results The sensitivities of PET imaging alone,thln layer CT alone,joint ima-ging diagnosis of SPN are 92.6% ,88.2% ,96.4% ,specificity are 73.3% ,57.1% ,85.7% ,accuracy percentage rate are 89.3% ,80.4% ,94.6% ,the positive predictive values are 92.6% ,91.8% ,96.4% ,and negative predictive val-ues are 84.6% ,61.5% ,92.3% ,respectively. Conclusion 18 F-FDG PET combined with thin layer CT scanning is a very effective method to clinical diagnosis the malignant degree of SPN.