[Objective] To use Platelet Additive Solution Ⅲ M to suspend concentrated platelets and evaluate their quality at different storage periods, in order to investigate the optimal ratio of Ⅲ M to plasma in the medium for storing concentrated platelets. [Methods] Disposable plastic blood bags with platelet storage bags were used to collect whole blood from healthy voluntary blood donors, and concentrated platelets were collected by plasma-rich method, with a volume of about 50 mL and ≥4.0×10¹⁰ platelets contained in each bag. According to the Platelet Addictive Solution ⅢM/plasma volume ratio in the medium of suspended platelets, the platelets were divided into 3 groups: control group (plasma only), experimental group 1(Platelet Addictive Solution ⅢM/plasma volume ratio of 6.5∶3.5) and experimental group 2 (low plasma group, Platelet Addictive Solution ⅢM/plasma volume ratio of 9∶1), each group of 50 samples. Three groups of platelets were stored at (22±2) ℃ at a flat-bed shaker, and 5 mL were sampled by sterile connection at day 1, 3, 5 and 7 respectively to detect platelet count, pH value, lactate dehydrogenase, CD62P positive rate and Annexin V positive rate. All the data were analyzed with SPSS24.0 software. One-way ANOVA was employed to compare the differences among three groups. In order to pairwise comparisons between means of multiple samples, Bonferroni method was applied. [Results] With the extension of storage time, the platelet count decreased and the Annexin V positive rate increased in the 3 groups, and the difference of the 3 groups was not statistically significant (P>0.05). The pH value decreased in the 3 groups, with values at day 1, 3, 5 and 7 of 7.44±0.13 vs 7.44±0.14 vs 7.41±0.11, 7.31±0.68 vs 7.43±0.23 vs 7.22±0.12, 7.30±0.15 vs 7.42±0.14 vs 7.17±0.12, 7.29±0.33 vs 7.26±0.18 vs 7.04 ± 0.12, respectively. The pH decline in the control group and experiment group 1 was minor, with no statistically significant difference, but experiment group 2 showed relatively larger decreases in day 5 and day 7, with statistically significant difference (P<0.05). LDH concentrate was elevated in 3 groups (mmol/L), with values at day 1,3,5 and 7 of 169.62±99.33 vs 105.80±150.71 vs 77.14±105.38, 225.10±112.86 vs 116.00±72.77 vs 94.42±88.74, 249.42±79.55 vs 119.00±53.51 vs 118.35±80.39, 253.34±86.95 vs 147.71±90.71 vs 124.68±128.68 respectively. Compared with the control group, the difference was statistically significant (P<0.05). Experimental group1 had the smallest increase; CD62P positive rate increased in 3 groups (%), with values at day 1, 3, 5 and 7 of 26.22±11.74 vs 23.48±12.48 vs 40.49±11.86, 41.29±8.36 vs 33.53±25.64 vs 50.42±22.36, 59.59±10.13 vs 36.39±23.10 vs 50.94±20.50, 72.92±15.44 vs 55.54±23.65 vs 61.89±18.82 respectively. Compared with the control group, the increase in experiment group1 was smaller, and the difference was statistically significant (P<0.05). [Conclusion] Platelet Addictive Solution ⅢM/plasma volume ratio of 6.5∶3.5 is superior to traditional plasma in maintaining platelet quality during the in vitro preservation period of platelets.

Objective To design a custom 3D printed monoblock acetabular implant for reconstructing PaproskyⅢA acetabular bone defects and to analyze the stress distribution,displacement,and clinical reliability of the implant and surrounding bone using finite element analysis(FEA).Methods Bilateral hip computed tomography(CT)data of a patient with PaproskyⅢA acetabular bone defects were collected.Models were developed and analyzed using Mimics Medical 21.0,Geomagic Wrap 2021,Solidworks 2023,and ANSYS Workbench 2022 R1 softwares.The biomechanical performance of the custom 3D printed monoblock acetabular implant was simulated under a single-leg stance condition.Results The peak von Mises stress of the hip components was observed at the femoral stem,measuring 67.318 MPa.For the custom 3D printeded monoblock acetabular implant,the peak stress was located at the anterosuperior contact area between the implant and acetabular bone,measuring 6.935 MPa.The femoral stem exhibited a peak stress of 67.318 MPa at its junction with the femoral head.The liner's peak stress was 1.333 MPa near the fixation of screw 9 at the superior part of the acetabular cup.The screws showed a peak stress of 2.215 MPa at the junction with the implant.For the cortical bone,the peak stress was 9.844 MPa at the distal femur,while the cancellous bone exhibited a peak stress of 0.701 MPa at its distal connection with the femoral stem.The pelvic bone's peak stress was 8.002 MPa at the anterior transition zone between the normal acetabulum and the defect.The peak micromotion of the custom 3D printed monoblock acetabular implant at its posterosuperior area,measuring 0.114 mm.The femoral stem and head exhibited a peak micromotion of 0.132 mm at the contact interface with the acetabular liner.The micromotion range at the implant-acetabular bone interface was 0.098 mm to 0.131 mm.Conclusion Under a simulated single-leg stance condition,the stress distribution in all components and the acetabular bone surface remains below their respective yield strengths.The micromotion threshold between the acetabular cup and acetabular bone is within acceptable limits.Biomechanical analysis indicates that the patient can perform early weight-bearing rehabilitation postoperatively.However,walking or jogging rehabilitation should be approached with caution.

BACKGROUND:Bone mineral density is the clinical gold standard for determining bone strength,but bone mineral density is less sensitive to changes in bone mass,with large changes in bone mineral density only occurring when bone mass is significantly reduced,so bone mineral density has limited ability to predict changes in bone strength and fracture risk. OBJECTIVE:A model of the normal and osteoporotic hip joint was developed to analyze the stresses and deformation in the hip of normal and osteoporotic patients under single-leg standing conditions. METHODS:A healthy adult female volunteer at the age of 36 years was selected as the study subject.The CT data of the hip joint of this volunteer were obtained and saved in DICOM format.The hip joint model was reconstructed in three dimensions,and the material properties were assigned by the gray value assignment method to obtain the normal and osteoporotic hip joint models according to the empirical formula.The same boundary conditions and loads were set to simulate the stresses and deformation in the normal and osteoporotic hip joints in the single-leg standing position. RESULTS AND CONCLUSION:(1)In the finite element model of the normal and osteoporotic hip,the stress distribution was more concentrated in the medial region of the femoral neck.(2)In the hip bone,the stress distribution was mainly concentrated in the upper part of the acetabulum.(3)The stress peaks in the medial femoral neck and upper acetabulum were larger in the normal hip model than in the osteoporotic hip model,probably due to the reduced bone strength of the osteoporotic bone.(4)The peak Von Mises of both normal and osteoporotic hip models were concentrated on the medial femoral neck,and the peak Von Mises of the hip bone was smaller,indicating that the overall effect of osteoporosis on hip bone stresses was relatively small.(5)In terms of deformation in the single-leg standing position,the maximum deformation in the normal hip model was located at the acetabulum and femoral head,and the maximum deformation was located at the upper part of the greater trochanter of the femur.(6)It is suggested that the finite element analysis method to model the values of parameters related to bone tissue in osteoporosis may improve clinical prediction of bone strength changes and fracture risk.It is explained from the biomechanical view that the intertrochanteric femur and femoral neck are good sites for osteoporotic hip fractures.

OBJECTIVE To investigate the transfer and diffusion of berberine hydrochloride(BBH), the main active component of Coptis and Phellodenticum in the system of carboxymethyl chitosan(CMCS)-sodium alginate(SA). METHODS CMCS and SA were stirred in a certain proportion, and D-gluconolactone(GDL) was added to form polyelectrolyte hydrogel. Rheometer was used to study the rheological properties of CMCS-SA hydrogel, including the elastic modulus G′ and the viscous modulus G′′. A BBH diffusion model for CMCS-SA hydrogel was designed, and the relevant rules of BBH permeation through CMCS-SA polyelectrolyte hydrogel were observed by UV-VIS. RESULTS The elastic modulus G′ of the hydrogel was measured when the ratio of CMCS to SA was 3∶1, 2∶1, 1∶1, 1∶2, and 1∶3. When the ratio of CMCS to SA was 1∶1, G′ was the highest, and the crosslinking strength of the hydrogel was the highest. The cumulative amount of BBH transfer was measured by the BBH transfer model, and the diffusion of BBH in CMCS-SA hydrogel was fitted as the skeleton dissolution by Peppas equation, indicating that BBH dissociation and the transfer efficiency increased as the amino group of CMCS decreased or the carboxyl group of SA increased. The elastic modulus G′ of CMCS-SA hydrogel increased with the increase of GDL content. The reason was that the binding force between CMCS and SA molecules gradually increased with the decrease of pH, and the crosslinking degree of the hydrogel was enhanced. When ratio of fixed CMCS to SA was 1∶1 and the GDL content was 0.15 g·mL−1, the formability of CMCS-SA hydrogel was good. In addition, when BBH was transferred in hydrogel with different concentrations of GDL, the transfer efficiency increased with the increase of GDL content. When BBH was delivered in different thickness hydrogel, the delivery efficiency of CMCS-SA hydrogel increased with the decrease of thickness. CONCLUSION The CMCS-SA hydrogel system, as a potential drug carrier for traditional Chinese medicine extracts such as BBH, is expected to serve as a gel carrier for transdermal drug delivery.

In order to understand the prevalence of Akabane disease(AKAD)in Guizhou Province and the molecular characteristics of the isolates,the whole-genome sequence of a strain of Akabane virus(AKAV)from a bovine AKAD-positive sample was determined and analyzed.The genotype and genetic variation of the strain were also explored.Based on the conserved S sequence,a fluores-cence quantitative PCR(qPCR)detection method was established and applied for the investigation of AKAV infection status in four large-scale beef cattle farms of Guizhou.Results showed that the S,M and L fragments of the bovine strain were highly homologous to the Tianjin strain(TJ2016/China/2016)and the Australian strain(JaLAB39/Australia/1959),where they were in the same evolutionary branch and belonged to genotype Ⅱ.Sensitivity assay found that the lowest detection limit was 2.5 X 101 copies/μL.Specificity assay showed the established method detected only AKAV with no amplification on bovine bluetongue virus(BLUV),Pasteurella multocida(PM),bovine infectious rhinotracheitis virus(IBRV)and bovine Mycoplasma bovis.The variation coefficients of inter-and intra batches in the repeatability test were both lower than 2.26％.These findings illus-trated that the established qPCR method had high sensitivity,good specificity and repeatability.A total of 298 serum samples from 4 large-scale beef cattle farms in Qianxi City and Huangping County of Guizhou Province were collected and tested for AKAV by the method.Out of 298 sam-ples,25 positive samples(25/298)were detected as positive with a positive rate of 8.39％.In sum-mary,this work provided the reference data for a deep understanding of the molecular prevalence of AKAV in Guizhou Province and laid foundation for the prevention and control of AKAD.

Alzheimer's disease(AD)is one of the most common types of dementia in clinical practice.The increased permeability of the gut and blood-brain barrier caused by intestinal microecological dysbiosis may be an important incentive to affect the incidence of AD.Gut microbiota regulates the central nervous system through the gut microbiota-gut-brain axis(gut-brain axis),and plays an important role in the cognitive func-tion,occurrence and development of clinical symptoms in AD patients.This review comprehensively reviewed the current literature on the relationship between gut microbiota dysregulation and inflammatory cytokine changes in AD,and the treatment of AD targeting with gut microbiota,so as to clarify the new progress in the influence of intestinal microecology changes on the cognitive function of AD patients,the potential diagnosis in the early onset of AD,and the potential mechanism of gut microbiota participating in the regulation of AD.It provides a new idea for the treatment strategy of AD.

【Objective】 To investigate whether the blood donors' coagulation status may lead to apheresis platelet aggregation in vitro. 【Methods】 Thirty blood donors with aggregation in apheresis platelets collected by AMICUS blood cell separator no less than 3 times previously and occurred when the last time of apheresis donation were observed in aggregated group (referred to as the experimental group); Thirty donors without aggregation in apheresis platelets collected by AMICUS blood cell separator no less than 3 times were observed in the control group simultaneously. The basic platelet parameters in the two groups, including Plt, MPV, PDW, Pet, P-LCR were detected by automatic blood cell analyzer (BC-3000Plus), and thromboelastogram indexes including reaction time(R), kinetics time(K), kinetics of clot development(α), maximum amplitude (MA) and coagulation index(CI) were tested by Thrombosis elastography (TEG) before collection. With SPSS24.0 software, t test was used to compare the differences between the two groups. 【Results】 The CI value in experimental group was significantly different from that of the control group (0.48± 1.00 vs -0.99 ±1.96, P< 0.05), and there was no significant difference in all above basic platelet parameters and other TEG parameters (P>0.05 ) . 【Conclusion】 The coagulation status of blood donors may be an independent risk factor for the in vitro aggregation of apheresis platelets.

Objective:To investigate the effect of 3D-printed customized flanged cup in hip revision with severe acetabular bone defects.Methods:Since February 2017, 10 cases of 3D-printed customized flanged cups were used in hip revision with severe acetabular bone defects, including 2 cases of Paprosky type IIIA and 8 cases of Paprosky type IIIB. There were 5 males and 5 females, mean age 73.6±8.1 yrs (range, 62-87 yrs), 5 left and 5 right cases. The preoperative thin-layer CT scan was preformed to reverse reconstruct digital pelvis. Five cases of one-piece flanged cups and 5 cases of decomposed flanged cups, including 3 cases of composite one-piece cups were designed by computer. The surgery was performed strictly according to the plan.Postoperative follow-up was performed to evaluate the Harris score. Operation time,intraoperative bleeding and other complications such as vascular and nerve injury, postoperative infection, and dislocation were counted. Pelvic X-ray was used to assess the height and horizontal position of the center of rotation and the stability of the prosthesis.Results:The surgical procedure was successful, with an average operative time of 147.9±48.3 min (range, 96-212 min) and an average intraoperative bleeding of 730.4±262.6 ml (range, 500-1 300 ml). The mean time of final follow-up was 40.8±18.7 months (range, 16-70 months) after surgery. At the last follow-up, the average Harris score was 83.80±6.73, with 4 cases excellent, 5 cases good, and 1 case fair. The excellent and good rate was 90%. The last Harris score was significantly higher than that before operation 28.60±8.40 ( t=16.84, P<0.001). The height of affected hip joint rotation center decreased from 46.24±7.74 mm before operation to 15.54±2.54 mm after operation with significant difference ( t=14.61, P<0.001). It was slightly higher than the opposite side (13.81±1.48 mm), which had no significant difference ( t=1.83, P=0.100). The horizontal distance of affected hip joint rotation center increased from 33.79±5.27 mm before operation to 40.53±4.50 mm after operation with significant difference ( t=3.62, P=0.006). It had no significant difference ( t=1.28, P=0.232) compared with the opposite side (38.54±3.46 mm). All incisions were healed in one stage without infection, vascular or nerve injury. During the following-up, all prostheses were in satisfied position without loosening, dislocation or screw breaks. Conclusion:Digitally assisted 3D-printed flanged cups can be used in hip revision with severe acetabular bone defect. It can not only improve hip joint function, but also restore the acetabular rotation center and the prosthesis stability, which can achieve good early and mid-term effect.

Objective:To analyze the efficacy of the reconstruction of the proximal humerus by reverse shoulder arthroplasty with three-dimensional (3D) printing technology after tumor rescetion.Methods:A retrospective analysis was conducted on the data of eight patients undergoing semi-constrained reverse shoulder arthroplasty with 3D printing technology after the resection of bone tumors in proximal humeri at the Affiliated Wuxi People's Hospital of Nanjing Medical University from December 2017 to January 2021. There were four males and four females with an average age of 55.1 (range, 31-73) years, all of whom had unilateral onset, 2 on the left and 6 on the right. There was one case of leiomyosarcoma (Enneking IIB), two cases of chondrosarcomas (one Enneking IA and one Enneking IB), four cases of Campanacci grade 3 giant cell tumor of bone, and one case of bone metastasis of lung adenocarcinoma. Individualized prosthesis and implantation protocol were completed preoperatively in all patients. The glenoid baseplate was manufactured using 3D printing technology. During the surgery, Malawer type I tumor resection and semi-constrained reverse shoulder arthroplasty were performed, with 6 cases simultaneously using allograft-prosthetic composite reconstruction. The follow-up was scheduled, and the patient received X-ray examination of the shoulder. The range of motion of the shoulder was measured, the Constant-Murley score and musculoskeletal tumor society (MSTS) score were recorded.Results:All 8 patients successfully completed the surgery, with a surgical time of 173.8±46.7 min (range, 130-260 min), intraoperative blood loss of 487.5±334.6 ml (range 200-1,200 ml), and proximal humeral resection of 9.9±4.6 cm (range, 4.5-19.0 cm). All patients were followed up for a period of 45.6±12.5 months (range, 24-60 months). At the last follow-up, the abduction ranges of motion of the affected shoulders increased from 27.5°±14.4° pre-operatively to 106.3°±21.8° post-operatively, with a statistically significant difference ( t=11.37, P<0.001). The forward flexion ranges increased from 28.1°±12.8° pre-operatively to 115.6°±24.0° post-operatively, with a statistically significant difference ( t=11.49, P<0.001). The Constant-Murley score was improved from 40.5±14.3 pre-operatively to 79.3±11.2 post-operatively, with a statistically significant difference ( t=9.58, P<0.001). The MSTS score was 25.6±2.2 (range, 23-28), including 6 excellent cases and 2 good cases. At 2 weeks after surgery, one patient experienced joint dislocation that was successfully reduced manually. Up to the final follow-up, all patients had survived without local tumor recurrence, metastasis, prosthesis infection and loosening. Conclusion:3D printing technology assisted shoulder arthroplasty is helpful for effective reconstruction and shoulder joint function recovery after resection of proximal humeral tumors, with satisfactory outcomes in the early and middle stages.

OBJECTIVE: Hypertrophic scars (HS) are a variety of skin tissue fibrosis disease that occurs in human skin, the effective therapeutic method of which is still inaccessible up to now. As a bioactive constituent of a well-known medical plant, Salvia miltiorrhiza (Danshen in Chinese), tanshinone II_A (TSA) is reported to inhibit cell proliferation in HS. Therefore, the aim of this study was to prepare TSA self-soluble microneedles to strengthen its dermal retention and break through the difficulty of significantly thickening epidermal connective tissue and stratum corneum at the HS site. The possible mechanism of action in suppressing HS was studied using human skin fibroblasts (HSF). METHODS: Tanshinone II_A self-dissolving microneedles (TSA-MN) was prepared using a negative mold casting method. The prescription process of microneedle was optimized by Box-Behnken effect surface method. Different media were selected to investigate the ability of transdermal absorption and in vitro release. Furthermore, according to Cell Counting Kit-8 (CCK8) method as well as the Western blot method, the effect of TSA-MN on the biological characteristics of HSF was investigated. RESULTS: With remarkable slow release effect and dermal retention, the release and transdermal properties of TSA-MN in vitro were better than both TSA and ordinary dosage forms. Its effect of HSF confirmed the essential decrease in cell motility during cell proliferation and cell migration in vitro, which plays a significant role in down-regulating the secretion of transforming growth factor-β₁ (TGF-β₁) in HSF and increasing the expression level of Smad7. CONCLUSION The prepared TSA self-soluble microneedles is helpful in solving the problem of hypertrophic scars, with a stable dermal retention effect after process optimization.