Building a novel TRUCK by harnessing the endogenous IFN-gamma promoter for cytokine expression.

Despite the remarkable success of CAR T therapy in hematological malignancies, its efficacy in solid tumors remains limited. Cytokine-engineered CAR T cells offer a promising avenue, yet their clinical translation is hindered by the risks associated with constitutive cytokine expression. In this proof-of-concept study, we leverage the endogenous IFN-γ promoter for transgenic IL-15 expression. We demonstrate that IFN-γ expression is tightly regulated by TCR signaling. By introducing IRES-IL15 into the 3'-UTR of the IFN-γ gene via HDR-mediated knock-in, we confirm that IL-15 expression can co-express with IFN-γ in an antigen-stimulation-dependent manner. Importantly, the insertion of transgenes does not compromise endogenous IFN-γ expression. In vitro and in vivo data demonstrate that IL-15 driven by the IFN-γ promoter dramatically improves CAR T cells' antitumor activity, suggesting the effectiveness of IL-15 expression. Lastly, as part of our efforts toward clinical translation, we have developed an innovative two-gene knock-in approach. This approach enables the simultaneous integration of CAR and IL-15 genes into TRAC and IFN-γ gene loci using a single AAV vector. CAR T cells engineered to express IL-15 using this approach demonstrate enhanced antitumor efficacy. Overall, our study underscores the feasibility of utilizing endogenous promoters for transgenic cytokines expression in CAR T cells.

Investigation of Novel Thermosetting Copolymer-Based Monomethylolurea-Glyoxal for Wood Manufacturing.

The purpose of this investigation was to design novel alternating copolymers (monomethylolurea-glyoxal, MMU-G) as adhesives for wood manufacturing. MMU-G were synthesized under acid (pH = 5) conditions. After the 120-day storage period, the MMU-G resins were used for plywood production, which exhibited a wet shear strength of about 2.15 MPa, similar to the freshly prepared MMU-G resin. The excellent water resistance and long storage stability showed that MMU-G has particular characteristics and properties all of their own, which, in certain respects, are very different from those of urea-formaldehyde (UF) adhesives. The X-ray diffraction results showed that only a few crystallinities occurred in MMU-G resins, indicating the presence of long side chains in the MMU-G polymer structures, leading to better adhesion strength than UF resins. The structure characteristics of the MMU-G resin were studied by Fourier transform infrared and electrospray ionization mass spectrometry, and a possible molecular structure has been inferred, which is consistent with spectroscopic results.

Study on Driving Factors and Spatial Effects of Environmental Pollution in the Pearl River-Xijiang River Economic Belt, China.

As a typical basin area in China, the Pearl River-Xijiang River Economic Belt (PRXREB) faces multiple types of environmental problems caused by the different development conditions of basins. To identify the situations of environmental pollution in the PRXREB, this paper constructed the Environment Pollution Composite Index (EPCI) by using four environmental pollutant emission indicators based on the entropy weight method, and explored the spatial effects and driving factors of environmental pollution by using the Spatial Error Model (SEM). The results showed that: (1) EPCI of the PRXREB decreased significantly from 2012 to 2016, and the spatial patterns were relatively stable. Wherein, the midstream and downstream were always the critical areas of environmental pollution. (2) Spatial spillover effects were significant in the PRXREB, which revealed that the local environmental pollution degree was affected by adjacent areas. (3) Industrial structure, infrastructure construction, and regulatory measures were the main driving factors of environmental pollution in the PRXREB. (4) To balance economic development and environmental protection in basin areas, environmental regulations such as environmental access, pollution payment, and cross-border early warning should be jointly established.

Wogonoside inhibits inflammatory cytokine production in lipopolysaccharide-stimulated macrophage by suppressing the activation of the JNK/c-Jun signaling pathway.

BACKGROUND: Mediated by innate immune cells, inflammation is an underlying presence in the pathogenesis of numerous pulmonary diseases. Macrophages play a critical role in mediating the initial response to infection in the lungs. When there is excessive activation of macrophages, hyper-production of inflammatory factors occurs, with inflammation as the ultimate result. Wogonoside, a bioactive flavonoid glycoside, has been reported to alleviate pulmonary inflammation. However, the mechanism underlying the anti-inflammatory effect of wogonoside has not yet been clarified. METHODS: The productions of nitric oxide (NO) and reactive oxygen species (ROS) were determined using a Griess reagent kit and a DAF-FM DA fluorescent probe, respectively. Moreover, the mRNA levels of inflammatory factors were quantified by qPCR, and the binding ability of c-Jun to promoters of inflammatory factors was performed by ChIP assay. Western blot was employed to detect the protein expression of inflammatory factors and signaling pathway. RESULTS: In this study, we found that pre-treatment with wogonoside dramatically suppressed lipopolysaccharide (LPS)-induced increase in the protein and mRNA levels of inflammatory factors in macrophages, such as cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), interleukin (IL)-1ß, tumor necrosis factor (TNF)-α, and IL-6. Furthermore, wogonoside profoundly reduced the increase in NO and ROS production and significantly blocked phosphorylation of JNK in LPS-stimulated macrophages. As revealed by Western blot and qPCR analysis, wogonoside mediated the JNK-dependent inhibitory effect. Compared with wogonoside alone, a combination of wogonoside and JNK inhibitor SP600125 provided no extra benefit in suppressing the protein expression and mRNA levels of inflammatory factors in LPS-stimulated macrophages. Additionally, ChIP analysis demonstrated wogonoside to remarkably reduce c-Jun enrichment in COX-2, iNOS, IL-1ß, TNF-α, and IL-6 promoters. CONCLUSIONS: Collectively, our findings showed that wogonoside notably suppresses LPS-stimulated production of inflammatory factors by repressing the activation of the JNK/c-Jun signaling pathway in macrophages. This suggests that wogonoside could serve as a promising therapeutic agent for pulmonary diseases related to macrophage inflammation.

[EFFECTIVENESS OF LOCKING COMPRESS PLATE FOR TREATMENT OF ASEPTIC DIAPHYSEAL HUMERAL NONUNIONS].

OBJECTIVE: To evaluate the effectiveness of locking compress plate (LCP) for the treatment of aseptic diaphyseal humeral nonunions. METHODS: Between January 2006 and January 2012, 23 patients with aseptic diaphyseal humeral nonuninons were treated with LCP and autologous iliac crest bone graft, and the clinical data were retrospectively analyzed. There were 15 males and 8 females with the average age of 42.5 years (range, 28-60 years). The fracture located at left side in 11 cases and right side in 12 cases. The mechanism of the injury was traffic accident in 15 patients, and falling from height in 8 patients. Fracture was treated by internal fixation in 20 cases and external fixation in 3 cases. And 6 patients had open fractures and other 17 had close fractures. Based on the Weber-Cech classification, 6 cases were rated as atrophic nonunions, and 17 cases as hypertrophic nonuninons. Shoulder function was evaluated by Constant-Murley score and elbow function was evaluated by Mayo score. RESULTS: After operation, 2 patients had transient radial nerve symptoms of numbness and 1 patient had superficial infection. Primary healing of incision was obtained in the other patients. All patients were followed up 22.22 months on average (range, 16-30 months). Normal range of motion of the shoulder was found in 11 cases; and limited movements of abduction, elevation, and posterior extension were observed in 12 cases. And osseous union was observed clinically and radiographically in all patients. The average union time was 16.95 weeks (range, 12-24 weeks). The average Constant-Murley score was 81.87 (range, 50-98); and shoulder function was excellent in 14 cases, good in 6, and fair in 3. And the average Mayo score was 87.78 (range, 70-96); and the result was excellent in 14 cases, good in 7, and fair in 2. CONCLUSION: Aseptic diaphyseal humeral nonunions can be successfully treated with LCP, coupled with the use of autologous iliac crest bone graft.

[Design of ABC damage variable and positioning system for acetabular fractures and 1122 cases multi-center statistic analysis].

OBJECTIVE: To design ABC damage variable and positioning system for acetabular fracture and explore the feasibility and clinical practical value of the system through the multi-center analysis of 1122 acetabular fractures. METHODS: According to acetabular three-column conception, and pelvic ring lesions damage direction caused by acetabular fracture domino effect and injury degree of proximal femur joint, it defined class A as any column acetabular fracture; class B as any two-column acetabular fracture; class C as front, dome and posterior mixture acetabular fracture. Lower case English letters a, m, p represented front, dome, posterior fracture, respectively. Acetabular damage variables: 1 was simple displaced fractures; 2 was comminuted fractures; 3 was compression fractures. Pelvic ring lesions damage variables: alpha was sacroiliac joints or sacroiliac fracture horizontal separation deflection; beta was sacroiliac joints or sacroiliac fracture vertical separation deflection; gamma was pubic symphysis separation/superior and inferior ramus of pubis fracture deflection; alpha beta gamma delta was compound floating damage. Proximal humerus joint damage variables: I was femoral head fracture; II was femoral neck fracture; II was intertrochanteric fractures of femur; IV was I to III compound fracture. The ABC damage variable positioning system for acetabular fracture was made up by the above-mentioned variables. The statistics from March 1997 to February 2010 showed 1122 cases acetabular fractures with 18 cases of double side acetabular fracture and 1140 cases of acetabular fractures. The pelvics anterior-posterior view, ilium and obturator oblique view, and 2/3D-CT materials were analyzed and researched. RESULTS: Each damage variables distribution situation in 1140 cases of acetabular fracture involved A in 237 cases (20.8%), B in 605 cases (53.1%), C in 298 cases (26.1%);front column fracture in 808 cases(70.9%), dome fracture in 507 cases (44.5%), posterior fracture in 1026 cases (90%). Acetabular variables: variabe 1 in 203 cases of simple displaced fracture (17.8%); variabe 2 in 516 cases of comminuted fracture(45.3%); variabe 3 in 421 cases of compression fracture (36.9%); 249 cases of pelvic ring lesions damage (21.8%), 75 cases femoral head fracture (6.6%); 18 cases of double side acetabular fracture and relative pelvic ring and proximal humerus joint variables (1.58%). Key part and curative effect elements of 1140 cases acetabular fracture: 507 cases of dome or posterior acetabular fracture (44.5%); 421 cases of compression fracture (36.9%); 249 cases of pelvic ring variables (21.8%); 75 cases of proximal humerus joint variables (6.6%); 486 cases of simple Aa/pl/2,Bapl/2 acetabular fracture (42.6% ). CONCLUSION: Compression fracture, especially defected compression fracture, takes important part in acetabular damage variables, and also presents that acetabular fracture with pelvic ring and proximal femoral damage variables are not rare at all. The relationship of the acetabular fracture damage variables, and its percentage shows the key points and elements in clinical treatment: weight-bearing to dome accounts for 44.5%; compression to defects account for 36.9%, pelvic ring to float accounts for 21.8%; dome fracture to double side fracture account for 6.6%. The system has significant guiding effects on clinic in terms of evaluation of injury severity, anatomic localization, difficulty index, alternative strategy, operative approach, effect of treatment,and prognosis. And the most important thing is that the system creates the comparison of damage variables in same type of fracture and the communication of homo-language and explores a new method.

The mechanical and biological properties of an injectable calcium phosphate cement-fibrin glue composite for bone regeneration.

Calcium phosphate cement (CPC) that can be injected to form a scaffold in situ has promise for the repair of bone defects. However, its low-strength limits the CPC to non-stress-bearing repairs. Fibrin glue (FG) with good sticking property and biocompatibility is possible used to reinforce the CPC. The objective of this study was to investigate the effects of FG on the mechanical and biological properties of CPC in an injectable CPC-FG composite. The initial setting time of this CPC-FG was delayed compared with the CPC control at different powder/liquid (P/L) mass ratio (p > 0.05). At a P/L of 5, the strength was (38.41 +/- 4.32) MPa for the CPC-FG, much higher than (27.42 +/- 2.85) MPa for the CPC alone (p < 0.05). SEM showed bone marrow stromal cells (BMSCs) with healthy spreading and anchored on the CPC-FG composite. After 14 days, the alkaline phosphatase (ALP) activity was (538 +/- 33) for the BMSCs on the CPC-FG and (517 +/- 27) for the BMSCs on the CPC alone. Both ALPs were higher than the baseline ALP (93 +/- 10) for the undifferentiated BMSCs (p < 0.05). The results demonstrate that this stronger CPC-FG scaffold may be useful for stem cell-based bone regeneration in moderate load-bearing orthopedic applications.

[A comparisive study between intramedullary interlocking nail and plate-screw fixation in the treatment of tibial shaft fractures].

OBJECTIVE: To evaluate the effects of tibial shaft fractures treatment with plate-screw or intramedullary interlocking nail. METHODS: Thirty-five fractures were treated with intramedullary interlocking nail. Tape A, B and C were 11, 18 and 6 cases respectively according to AO classification. Forty-five fractures were treated with plate-screw system. Type A,B and C were 10,22 and 13 cases respectively. The mean time between injury and operation was 3 days and 3.5 days respectively. The patients were evaluated with operation time,range of motion,time of bone union,and complications after a mean followup of 13 months(range 8 to 26 months). RESULTS: The mean operation time and the mean range of motion of the ankle and knee were found similar in both groups. The average dorsiflexion angle of ankle was 13 Degrees (0 degrees to 20 degrees) in locked intramedullary nailing group and 11 degrees (0 degree to 20 degrees) in plate group. The average plantar flextion angle of ankle was 41 degrees (30 degrees to 50 degrees) in locked intramedullary nailing group and 47 degrees (30 degrees to 50 degrees) in plate-screw group. The mean time of bone union was 3.3 months with intramedullary nailing and 3.5 months with plate-screw fixation. Length discrepancy occurred in only two patients (2 and 2.5 cm) with intramedullary interlocking nail. One patient treated by intramedullary interlocking nail for a type III open fracture occured osteomyelitis. Plain radiographs showed rotation in two patients with intramedullary interlocking nail and angulation in 3 patients with plate-screw fixation, which were distal-third tibial fractures. CONCLUSION: Plate-screw osteosynthesis could attain satisfactory results in uncomminuted tibia shaft fractures, and locked intramedullary nailing is more appropriate in comminuted fractures.

[A method to avoid the fixator failure by using pedicle screw combined vertebroplasty for spine fractures].

OBJECTIVE: To study a new implant material (carbonated hydroxyapatite, CHA) united pedicle screw to cure spine fracture. METHODS: Thirty-two cases of spine compressed fracture were used with pedicle screw fixator and vertebroplasty. Before operation, patients' vertebral body were compressed (46 + 21)% (20% approximately 70%) on average. In operation, broken vertebral body was reposition through pedicle screw technique, then used self-made syringe to inject CHA into anterior and central column of broken vertebral body through pedicle. And all of patients were not given any bone-graft. RESULTS: In 6 - 26 months followed-up, no immunologic rejection was found about hydroxyapatite, and no any broken of the screws and shafts was found, no loosing and other complications either. All the patients could move in 3 - 5 days after operation. The height of the broken vertebral body were reduced 97% compared with pre-operation. And CHA in vertebral body was degraded gradually, and at the same time it was replace by new bone in vertebral body. After operation, VAS score was 61 +/- 32, and there was significant difference compared with pre-operation. CONCLUSIONS: The pedicle screw fixation united vertebroplasty is an efficient way to prevent the failure of the treatment of spine fracture.

[Minimally invasive technique treatment of postoperative recurrent bone cysts in juvenile patients].

OBJECTIVE: To study the effect of minimally invasive treatment of postoperative recurrent bone cysts in juvenile patients. METHODS: From December 1984 to December 2003, 36 cases of postoperative recurrent bone cysts after focal curettage and bone graft included 19 males and 17 females, aging 9-21 years with an average of 15 years. The size of bone cyst ranged from 2.5 cm x 6.0 cm to 3.5 cm x 13.0 cm with an average of 3.0 cm x 8.0 cm. The locations were proximal humerus in 18 cases, humeral shaft in 10 and femoral trochanteric region in 8. The focal curette and bone graft were given once in 23 cases, twice in 10 cases and 3 times in 3 cases. The interval between recurrence and minimally invasive treatment was 5-13 months (6.5 months on average). The posteroanterior and lateral X-ray films were taken to determine the location, range and feature of the focus. Under local anesthesia, 2 canulated needles were used; one was used to aspirate the contents of the cyst, the other was used to inject hydrocortisone acetate. The dose was determined according to the range of the focus. The treatment was repeated every 3. 5-4 months until the focus healed. RESULTS: All patients were followed up from 3 to 18 years with an average of 5 years. The minimally invasive treatment was repeated 3-11 times with an average of 6 times. Twenty-six cases healed completely, 6 cases healed significantly, and 4 cases healed partially. No local or general complications occurred during the treatment. CONCLUSION: This minimally invasive method for the treatment of postoperative recurrent bone cyst in juvenile patients has following advantages : less pain, easy manipulation, no hospitalization, low cost and definite effect.

[Resection and reparation of heel with malignant melanoma].

OBJECTIVE: To investigate the surgical resection and reparation of heel with malignant melanoma. METHODS: Eight patients with malignant melanoma were treated from May 2001 to December 2003. The patients included 5 males and 3 females, and their ages ranged from 28 to 56 years. All lesions were located in the heel and were proved by pathological examination. According to Breslow classification, there were 2 cases of Grade I, 5 cases of Grade I, and 1 case of Grade II. Local extensive resection was performed in all cases. Lateral pedal skin flap, plantar medial artery island skin flap, and retrograde skin flap supplied by sural nutrition blood vessel were respectively applied in the reparation according to the size of heel soft tissue defect. The treatment with interferon was delivered before and after the operation. RESULTS: The surgical reparation was successful in all 8 cases. The postoperative follow-up was conducted from 18 months to 4 years. All patients remained alive and no tumor recurrence was observed. Considering the recovery of the function and sense, the best result was acquired with plantar medial artery island skin flap and lateral pedal skin flap, good with retrograde skin flap supplied by sural nutrition blood vessel. CONCLUSION: Local extensive resection is essential for the heel with malignant melanoma. Reparative reconstruction should be made on negative operative margin. Satisfactory clinical outcome is achieved by using lateral pedal skin flap, plantar medial artery island skin flap, and retrograde skin flap supplied by sural nutrition blood vessel.

[Clinical research of hemiprosthesis arthroplasty for the treatment of unstable intertrochanteric fractures in elderly patients].

OBJECTIVE: To report the clinical results of hemiprosthesis for unstable intertrochanteric fractures in elderly patients. METHODS: 86 cases (male 35 and females 51) of unstable intertrochanteric fractures in elderly patients were treated with hemiprosthesis arthroplasty in our hospital. The average age was 87 years old (range from 80 to 98 years old). According to the Evans classification of intertrochanteric fractures, there were 31 cases in type IIIA, 35 cases in IIIB and 20 cases in type IV. Most of the patients fell in the room and fractured, with concomitant diseases. After admitted to the hospital, the patients were consulted and treated for the concomitant diseases, and after the general conditions got stable and no absolute contraindications were found, the hemiprosthesis arthroplasty was performed. Almost all of the patients were operated on within 3 days with general anesthesia under the multi-function monitoring. The greater trochanter was reconstructed with figure of 8 wire fixation, the lesser trochanter with figure of 8 wire fixation, the lesser trochanter was fixed with circular wire around proximal femur. The defect within the femoral calcar was filled with bone cement and in a part of the patients, the intertrochanteric fracture was transformed into femoral neck fracture and the femoral calcar was fixed with femoral prosthesis stem. RESULTS: All operations were successful and the duration of the operation was 45 to 70 minutes with an average of 55 minutes. Blood was transfused in 73 patients during the operation between 400-800 ml with an average 450 ml. The patients got out of the bed within 3 days in 33 cases, 4-7 days in 43 cases, more than 8 days in 8 cases, and 2 patients couldn't get out of the bed. The period of hospitalization was between 13 to 27 days with an average of 16.8 days. COMPLICATIONS: 2 patients got slight deep phlebitis, 5 patients showed psychological disorders, and disorders in bacterial spectrum appeared in 3 patients, and after appropriate treatments, the complications gradually recovered and had no effect on the function of the affected extremity. 2 cases died of multiple organ failure during the hospitalization. 59 patients were followed up for 6 to 48 months with an average 16 months. The result was excellent in 17 cases who resumed their activities as before they got fractured, good in 35 cases who could walk with walking aid, fair in 5 cases who could walk with others help, and poor in 2 cases who could not walk. 1 patient died of other diseases 5 months after discharged. No dislocation, loosening or late infections occurred. 88.1% showed excellent or good result. CONCLUSION: It's an effective method to treat the unstable intertrochanteric fractures in elderly patients with hemiprosthesis arthroplasty. The result was satisfactory. It can decrease the complications, reduce the mortality, improve the patient's the living quality, and reduce the burden of the patient's family.