Objective:To evaluate the application of the reaming-irrigation-aspirator (RIA) technique in autologous bone grafting for non-union of lower limb fracture.Methods:A retrospective analysis was conducted of the data of 16 patients with non-union of lower limb fracture who had undergone autologous bone grafting using the RIA technique at Department of Lower Limb Orthopaedics Ⅰ, Zhengzhou Orthopaedic Hospital from July 2023 to March 2024. There were 12 males and 4 females with an age of (47.2±14.3) years. Sites of non-union: the femur in 14 cases and the tibia in 2 cases. Initial surgical fixation: intramedullary nail fixation in 5 cases, plate fixation in 10 cases, and intramedullary nail plus plate fixation in 1 case. The time for fracture non-union was (21.0±12.0) months. Intraoperatively, autologous bone reaming was conducted using a drill bit of RIA system, and simultaneous flushing using irrigation fluid. The bone debris carried out by the irrigation fluid was collected into a filtration device through negative pressure aspiration. After the filter was opened to squeeze out congestion, bone paste was taken out for standby. Internal fixation methods were adjusted according to the non-union condition of the fractures. Finally, the bone paste was implanted into bone defects. The time for autologous bone harvest, amount of bone harvested, intraoperative blood loss, postoperative pain at a donor site, fracture healing, functional recovery of a lower limb and occurrence of complications were recorded.Results:In 2 of the 16 patients, fracture of the RIA drill bit occurred when bone marrow was harvested from the tibia, and their operation was completed only after the drill bits were replaced. In the 16 patients, time for bone harvest was (31.4±6.1) min, bone harvest volume (42.6±9.5) mL, and intraoperative blood loss (815.6±238.6) mL. The incisions in this cohort healed primarily after operation, and 4-week follow-up revealed that no patients had pain symptoms at a donor site. The 16 patients were followed up for (8.0±2.0) months after operation. All patients achieved bone union after (4.4±0.5) months. Follow-ups showed that no patients had complications like delayed infection, donor site fracture, or loosening or fracture of an internal fixator. According to the rating scale for lower limb function at the last follow-up, the function of affected limb was evaluated as excellent in 7 cases, as good in 8 cases, and as fair in 1 case.Conclusion:Application of RIA technique for autologous bone grafting to treat non-union of lower limb fracture can lead to fine fracture healing and good lower limb function, but no long-term pain symptoms in the donor area for the patients.

Objective:To evaluate the application of the reaming-irrigation-aspirator (RIA) technique in autologous bone grafting for non-union of lower limb fracture.Methods:A retrospective analysis was conducted of the data of 16 patients with non-union of lower limb fracture who had undergone autologous bone grafting using the RIA technique at Department of Lower Limb Orthopaedics Ⅰ, Zhengzhou Orthopaedic Hospital from July 2023 to March 2024. There were 12 males and 4 females with an age of (47.2±14.3) years. Sites of non-union: the femur in 14 cases and the tibia in 2 cases. Initial surgical fixation: intramedullary nail fixation in 5 cases, plate fixation in 10 cases, and intramedullary nail plus plate fixation in 1 case. The time for fracture non-union was (21.0±12.0) months. Intraoperatively, autologous bone reaming was conducted using a drill bit of RIA system, and simultaneous flushing using irrigation fluid. The bone debris carried out by the irrigation fluid was collected into a filtration device through negative pressure aspiration. After the filter was opened to squeeze out congestion, bone paste was taken out for standby. Internal fixation methods were adjusted according to the non-union condition of the fractures. Finally, the bone paste was implanted into bone defects. The time for autologous bone harvest, amount of bone harvested, intraoperative blood loss, postoperative pain at a donor site, fracture healing, functional recovery of a lower limb and occurrence of complications were recorded.Results:In 2 of the 16 patients, fracture of the RIA drill bit occurred when bone marrow was harvested from the tibia, and their operation was completed only after the drill bits were replaced. In the 16 patients, time for bone harvest was (31.4±6.1) min, bone harvest volume (42.6±9.5) mL, and intraoperative blood loss (815.6±238.6) mL. The incisions in this cohort healed primarily after operation, and 4-week follow-up revealed that no patients had pain symptoms at a donor site. The 16 patients were followed up for (8.0±2.0) months after operation. All patients achieved bone union after (4.4±0.5) months. Follow-ups showed that no patients had complications like delayed infection, donor site fracture, or loosening or fracture of an internal fixator. According to the rating scale for lower limb function at the last follow-up, the function of affected limb was evaluated as excellent in 7 cases, as good in 8 cases, and as fair in 1 case.Conclusion:Application of RIA technique for autologous bone grafting to treat non-union of lower limb fracture can lead to fine fracture healing and good lower limb function, but no long-term pain symptoms in the donor area for the patients.

COVID-19 is caused by a novel coronavirus-severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), which has being spreading around the world, posing a serious threat to human health and lives. Neutralizing antibodies and small molecule inhibitors for virus replication cycle are the main antiviral treatment for novel coronavirus recommended in China. To further promote the rational use of antiviral therapy in clinical practice, the National Center for Infectious Diseases (Beijing Ditan Hospital Capital Medical University and the First Affiliated Hospital, Zhejiang University School of Medicine) invited experts in fields of infectious diseases, respiratory and intensive care to develop an Expert Consensus on Antiviral Therapy of COVID-19 based on the Diagnosis and Treatment Guideline for COVID-19 ( trial version 10) and experiences in the diagnosis and treatment of COVID-19 in China. The consensus is concise, practical and highly operable, hopefully it would improve the understanding of antiviral therapy for clinicians and provide suggestions for standardized medication in treatment of COVID-19.

Objective:A high performance liquid chromatography (HPLC) method was developed to determine the enzymatic activity of CD38 in blood, which was the major enzyme responsible for consuming nicotinamide adenine dinucleotide (NAD). Additionally, the study aimed to detect the differences in CD38 enzymatic activity among individuals of varying ages and health statuses.Methods:A 50 μl whole blood matrix and enzyme reaction substrate of 150 μl β-NAD at a concentration of 500 μmol/L were selected for the analysis. To eliminate the impact of endogenous β-NAD, the whole blood sample was pre-incubated at 37 ℃ for 20 minutes before adding the substrate. The reaction was terminated by perchloric acid (PCA) after incubation at 37 ℃ for 40 min. The change in product nicotinamide (NAM) before and after the enzymatic reaction was measured by HPLC to calculate the CD38 activity. The linearity, limit of detection, limit of quantification, precision, and stability of the method were evaluated. The CD38 enzymatic activities in 60 healthy volunteers and 30 colorectal cancer patients in blood were determined by the developed method.Results:Pre-incubation at 37 ℃ for 20 minutes eliminated the effect of endogenous β-NAD. The correlation coefficient of NAM was 0.999 in the concentration range of 0.1-3.2 μmol/L, with limit of detection of 0.5 nmol/L and limit of quantification of 2.1 nmol/L. The average within-run imprecision ( CV) and total CV were 3.22%-4.03% and 2.91%-4.70%, respectively. The recovery rate ranged from 94.82% to 96.81%. The CD38 activity of whole blood was stable by storage at 4 ℃ for 48 hours, storage at room temperature for 8 hours, thawing of frozen whole blood at room temperature for 2 hours, or repeated freeze-thawing three times. NAM, NAD standards, and pre-treatment samples were stable after 48 hours at 4 ℃ and 8 hours at room temperature. CD38 activity gradually decreased with increasing concentration of the added CD38 inhibitor 4-aminoquinoline derivative (78c). Measurement of 60 healthy physical examination population samples showed significantly higher CD38 enzyme activity in the elderly group than that in the young group ( t=-2.776, P=0.007) and measurement of 30 colorectal cancer patients showed significantly higher CD38 enzyme activity than that in healthy people ( t=-2.572, P=0.012). Conclusion:The established HPLC method for determining CD38 enzymatic activity is characterized by its simplicity, efficiency, accuracy, and reproducibility. This technique serves as a valuable tool for investigating aging and aging-related diseases.

Objective To evaluate the validity and security of our self-designed bone marrow stem cell Screen-Enrich-Combine(-Biomaterials) Circulating System(SECCS) for bone non-union of limbs. Methods From May 2013 to December 2015, 24 patients with limb non-union were treated at our de-partment. They were aged from 20 to 65 years (mean, 42.8 years). Their non-union involved femur in 17 cases, tibia in 4, radius in one, humerus in one and fibula in one. In surgery, 80 mL bone marrow blood was aspirated from the anterior superior iliac spine for rapid preparation of bone substitute(β-TCP)composite with bone marrow stem cells by SECCS which was then implanted at the non-union locations. The bone marrow blood was collected before and after enrichment for stem cell counts. The bone union, clinical union time and related com-plications were evaluated by follow-up X-rays at 1, 3, 6 and 9 months after surgery. Results Each collection of bone marrow blood took 13.5 minutes on average. The enrichment rate of stem cells was 81.2%. 11,751 ± 1,011 stem cells were implanted per patient on average. All the patients were followed up for 9 to 48 months (mean, 19.2 months). Twenty-two patients acquired clinical union 9 months after operation and the other 2 suffered from malunion due to insufficient bone implant, yielding a union rate of 91.6% and an average union time of 6.5 months. No graft infection or internal fixation failure occurred and no severe complications hap-pened at the donor or implant sites.Conclusion The bioactive bone substitute manufactured by our self-designed SECCS can be used as a novel therapy for limb non-union, and this device is moreover charac-terized with convenience, limited invasion and satisfactory osteogenesis so that complications of autologous bone transplantation can be avoided.

Objective To introduce a new method for preparation of bioactive β-tricalcium phosphate (β-TCP) by rapid stem cell screen-and-enrich-and-combine circulating system (SECCS) and evaluate its efficacy in the treatment of fresh fractures and bone defects.Methods Twenty-two patients with fresh fracture and bone defects were treated with SECCS from July 2013 to April 2016.They were 16 males and 6 females with an average age of 52.2 years (from 27 to 81 years).There were 15 tibial plateau fractures and 7 calcaneal fractures.The average size of bone defects was 12.5 mL.Bioactive β-TCP was prepared by SECCS intraoperatively and implanted back immediately into the bone defects.Radiographic examination,Lysholm knee scoring and Maryland foot scoring were used for assessment of curative efficacy.Results The 22 patients were followed up for an average of 25.7 months (from 12 to 46 months).By SECCS,the enrichment efficiency of bone marrow stromal cells (BMSCs) reached up to 82.4％ and the cell viability was not affected.The tibial plateau fractures were re-transplanted with 13,381.3 BMSCs on average and healed after an average of 8.9 weeks (from 6 to 15 weeks).The Lysholm knee scores at one year postoperatively averaged 93.6 points (from 84 to 100 points),yielding 10 excellent cases,4 good cases and one fair case.The calcaneal fractures were implanted back with 16,677.7 BMSCs on average and healed after an average of 9.4 weeks (from 8 to 13 weeks).The average Maryland foot score at one year after operation was 93.6 points (from 85 to 98 points),yielding 6 excellent cases and one good case.Conclusion Bioactive materials prepared by SECCS are good bone grafts for fresh fractures and bone defects.

Objective To investigate the clinical value of conventional ultrasonographic ( US),US-guided diffuse optical tomography ( US-guided DOT ) and both combined to assess treatment response of breast cancer to neoadjuvant chemotherapy( NAC). Methods Eighty-eight breast cancer patients,totally 93 lesions were included in the study. Pre-and post-last chemotherapy,size,and total hemoglobin concentration ( THC) of each lesion were measured by conventional US and US-guided DOT before biopsy,the change of lesion Size( ΔSize) and the change of THC( ΔTHC) were calculated respectively.Based on the guidelines to evaluate the response to treatment in solid tumors,the responses to NAC were classified into complete response, partial response, static disease, progressive disease groups. The histological response to chemotherapy were categorised as partial pathological response and complete pathological response using Miller and Payne system. Results Of 93 breast cancers,the overall response rate was 81.7%,the cPR rate was 24.7%. According to ROC curve analysis,when ΔSize 42.6% and ΔTHC 23.9% as cutoff values to evaluate the complete response and partial response,the area under the curve ( AUC ) were 0.666 and 0.751,respectively,the AUC of US and US-guided DOT combined in parallel and in series were 0.680 and 0.737 respectively.When ΔSize 64.5% and ΔTHC 27.2% as cutoff values to evaluate complete pathologial response,the AUC were 0.690 and 0.728 respectively,the AUC of US and US-guided DOT combined in parallel and in series were 0.693 and 0.726 respectively. Conclusions US-guided DOT and US can be used to predict breast cancers response to NAC,US and US-guided DOT combined in parallel and in series can not improve response prediction comparing with US or US-guided DOT alone.

Objective To evaluate the efficiency of our self-designed enriched filtrator of stem cells in screening and collecting enriched human bone marrow stem cells (BMSCs) for rapid construction of bioactive materials.Methods Bone marrow blood from 32 patients was circulated in our self-designed enriched filtratorand filtered through beta-tricalcium phosphate(β-TCP).The numbers of BMSCs were counted pre-and post-filtration to assess the enrichment efficiency.The numbers of nucleated cells,red blood cells and platelets were compared pre-and post-filtration to assess the effects of the enriched filtrator of stem cells on blood cells.Bacteriological examinations were performed to test the safety of the device.Scanning electron microscopy was carried out to observe the adhesion of BMSCs on β-TCP.Results There was a significant difference between the pre-fihration colony number of BMSCs cultured (232.8 ± 128.6/mL) and the post-filtration one (37.8 ± 34.9/mL) (t =10.743,P ＜ 0.001).The efficiency of BMSCs enrichment of the device was 85.6％ ±8.4％ (from 63.8％ to 97.1％).There were no significant differences between pre-filtration and post-filtration in the numbers of nucleated cells [(16.8 ± 5.4) × 109/mL versus (16.3 ±5.0) ×l09/mL],red blood cells [(3.9±0.6) ×1012/mL versus (3.8±0.6) ×1012/mL] and platelets [(120.4 ± 54.6) × 109/mL versus (123.0 ± 51.8) × 109/mL] (P ＞ 0.05).All the bacteriological examinations of the bone marrow samples were negative.After in vitro osteogenic induction culture,formation of numerous BMSCs colonies was observed on the β-TCP by scanning electron microscopy.Conclusion Since our self-designed enriched filtrator can efficiently collect enriched BMSCs and combine them with β-TCP simultaneously,the device is capable of rapid and safe construction of bioactive materials.

Objective To investigate the value of ultrasound-guided diffuse optical tomography (US-guided DOT) to predict clinical efficacy of breast cancer neoadjuvant chemotherapy (NAC).Methods Eighty-eight breast cancer patients with 93 lesions were included.Pre-and post-last chemotherapy,the size,total hemoglobin concentration (THC) of each lesion were measured by ultrasonography (US) and US-guided DOT.Based on the guidelines to evaluate the response to treatment in solid tumors,the lesions of treated breast cancer patients were divided into 4 types of responses to NAC:complete response (CR),partial response (PR),stable disease (SD),and progressive disease (PD).Efficient groups include CR and PR groups.Results As expected,no significant difference was found in size and THC for untreated lesions (all P ＞0.05).However,for the treated lesions(P =0.001),THC,pre-vs post-treatment size changes (△Size％) (P =0.002) and THC changes (△THC％) (P ＜0.001) were significantly varied among CR,PR,SD,PD groups.When compared with pre-treated,tumor sizes after treatment were changed significantly in all CR(P ＜0.001),PR(P ＜0.001),SD (P =0.023) and PD (P =0.001),while significant change of THC was only found in CR(P ＜0.001),PR(P ＜0.001) and SD (P =0.002).When △THC％ =23.9％ as the threshold for prediction of NAC efficiency,the area under the curve of ROC was 0.75,and the sensitivity was 73.7％,specificity was 76.5 ％,positive predictive value was 93.3 ％,negative predictive value was 39.4％,accuracy was 74.2％.Conclusions △size％ changes in consistent with △THC％ among the intergroups,but their changes levels are different,the highest change percent appears in CR,gradually decreased in PR,SD groups.△THC％ will contribute to predict preoperative clinical NAC efficacy.

Objective To investigate the relationship of greyscale ultrasonographic signs and clinical pathological characteristics with axillary lymph node metastasis in invasive breast cancer.Methods Three hundred and thirty-five patients with pathologically confirmed breast cancer were retrospectively analyzed,which categorized into 2 groups,including lymph node metastasis group and lymph node non-metastasis group.The clinical and pathological characteristics included age,primary tumor size,histological grade,and molecular subtypes.The ultrasonographic features were examined including orientation,shape,margin,echo pattern,posterior acoustic feature,and calcification.Chi-Square test and Logistic regression analysis were performed to analyze the relationship of clinical pathological characteristics and greyscale ultrasonographic signs with axillary lymph node metastasis.Results Age (＜45 years),primary tumor size (≥2 cm),histological grade(Ⅲ grade),molecular subtype (Luminal B),and number of margin angulation (＞ 10) contributed to axillary lymph node metastasis in 335 cases,according to univariate analysis (P ＜0.05).Multivariate Logistic regression analysis showed age (OR =0.573,P =0.019),the primary tumor size (OR=2.359,P =0.001),histological grade (OR=0.529,P =0.008),and number of marginangulation (OR =1.889,P =0.031) were risk factors of axillary lymph node metastasis in breast cancer.Conclusions Univariate and multivariate analyses show the correlation between ultrasonographic features and axillary lymph node involvement.Combining with clinical pathological data,it can provide a predictor of axillary lymph node metastasis.