OBJECTIVES: To characterize the biological properties of double-negative T (DNT) cells isolated from leukoreduction filter residues. METHODS: Leukoreduction filters containing residues from 400 mL whole blood units (n=6) were collected from a blood center. Filters were back-flushed with normal saline, and the eluate was concentrated to obtain leukoreduction filter residues. Leukocytes in the residues were counted by dual-fluorescence staining. DNT cells were then isolated from the residues using antibody-mediated adsorption and density gradient centrifugation. Both cryopreserved and fresh unstimulated DNT cells derived from the residues were subjected to in vitro culture. Following culture, cells were assessed for expansion fold, viability, immunophenotype, differentiation status, and cytotoxicity against target cells using dual-fluorescence staining and flow cytometry, with comparisons made to DNT cells derived from whole blood. RESULTS: The leukocyte recovery rate achieved through reverse flushing of the leukocyte reduction filter was (41.9±14.7)%. Compared to whole blood, the DNT cell starting material obtained from filter residues showed no significant difference in total T-cell content (P>0.05). However, the viability and purity of the resulting DNT cell starting materials were significantly lower (both P<0.05). After 17 days of culture, DNT cells from filter residues and whole blood showed no significant differences in expansion fold, immunophenotype, differentiation status, or cytotoxicity toward target cells (all P>0.05). However, the viability of DNT cells from residues was significantly lower than that of whole blood-derived DNT cells [(86.0±4.2)% vs. (92.2±1.2)%, P<0.05]. After thawing (post 3 or 15 days of cryopreservation) and 17 days of culture, DNT cell starting materials from residues showed comparable immunophenotype, expansion fold, and differentiation status to their non-cryopreserved counterparts from the same source (all P>0.05). However, the viability of DNT cells cryopreserved for 3 days [92.4% (91.8%, 92.8%)] and the cytotoxicity against target cells of those cryopreserved for 15 days [91.3% (89.4%, 95.1%)] were significantly higher than those of non-cryopreserved DNT cells [87.8% (82.0%, 89.0%) and 70.9% (67.3%, 80.2%), respectively] (P<0.05). CONCLUSIONS DNT cells derived from leukoreduction filter residues exhibited highly comparable characteristics to those from whole blood in terms of expansion, purity, differentiation, and biological potency. Furthermore, their biological activity post-cryopreservation and revival remained largely similar to non-cryopreserved cells. These findings suggest that leukoreduction filter residues represent a promising alternative source of starting material for manufacturing off-the-shelf, allogeneic DNT cell therapeutics.

The prevalence of hepatitis B represents a significant public health concern with a heavy disease burden.In China,there is still a big gap between the current diagnosis and treatment rates of hepatitis B and the goal of eliminating viral hepatitis as a public health threat by 2030 set by the World Health Organization(WHO).In order to achieve the WHO goal and the goal of 2030 Healthy China Outline,the Chinese Preventive Medicine Association organized domestic experts in the fields of clinical medicine,public health and clinical laboratory medicine to develop the Expert Recommendations on Screening,Testing and Management for Hepatitis B Virus Infection in Adults after several rounds of discussion based on comprehensive review of relevant domestic and international guidelines and literatures,the purpose is to facilitate universal screening of hepatitis B virus(HBV)infection in adults and provide practical guidance on disease assessment,treatment and long-term follow-up management of people infected with HBV and vaccination for people susceptible to HBV infection,thus promoting the elimination of the threat of hepatitis B.

Objective:To investigate the interactive effect of mismatch repair (MMR) protein status and adverse clinicopathological features on prognosis of stage Ⅰ-Ⅲ colon cancer.Methods:The retrospective cohort study was conducted. The clinicopathological data of 1 650 patients with colon cancer of stage Ⅰ-Ⅲ who were admitted to 7 hospitals in China from January 2016 to December 2017 were collected. There were 963 males and 687 females, aged 62(53,71)years. Patients were classified as 230 cases of MMR deficiency (dMMR) and 1 420 cases of MMR proficiency (pMMR) based on their MMR protein status. Observation indicators: (1) comparison of clinicopathological characteristics between patients of different MMR protein status; (2) analysis of factors affecting the survival outcomes of patients of dMMR; (3) analysis of factors affecting the survival outcomes of patients of pMMR; (4) interaction analysis of MMR and adverse clinicopathological features on survival outcomes. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test or Fisher exact probability. Comparison of ordinal data was conducted using the Mann-Whitney U test. The random forest interpolation method was used for missing values in data interpolation. Univariate analysis was conducted using the COX proportional risk regression model, and multivariate analysis was conducted using the COX stepwise regression with forward method. The coefficient of multiplication interaction effect was obtained using the interaction term coefficient of COX proportional risk regression model. Evaluation of additive interaction effects was conducted using the relative excess risk due to interaction ( RERI). Results:(1) Comparison of clinicopathological characteristics between patients of different MMR protein status. There were significant differences in age, T staging, the number of lymph node harvest, the number of lymph node harvest <12, high grade tumor between patients of dMMR and pMMR ( P<0.05). (2) Analysis of factors affecting the survival outcomes of patients of dMMR. Results of multivariate analysis showed that T staging, N staging, the number of lymph node harvest <12 were independent factors affecting the disease-free survival (DFS) of colon cancer patients of dMMR ( hazard ratio=3.548, 2.589, 6.702, 95% confidence interval as 1.460-8.620, 1.064-6.301, 1.886-23.813, P<0.05). Age and N staging were independent factors affecting the overall survival (OS) of colon cancer patients of dMMR ( hazard ratio=1.073, 10.684, 95% confidence interval as 1.021-1.126, 2.311-49.404, P<0.05). (3) Analysis of factors affecting the survival outcomes of patients of pMMR. Results of multivariate analysis showed that age, T staging, N staging, vascular tumor thrombus were independent factors affecting the DFS of colon cancer patients of pMMR ( hazard ratio=1.018, 2.214, 2.598, 1.549, 95% confidence interval as 1.006-1.030, 1.618-3.030, 1.921-3.513, 1.118-2.147, P<0.05). Age, T staging, N staging, high grade tumor were independent factors affecting the OS of colon cancer patients of pMMR ( hazard ratio=1.036, 2.080, 2.591, 1.615, 95% confidence interval as 1.020-1.052, 1.407-3.075, 1.791-3.748, 1.114-2.341, P<0.05). (4) Interaction analysis of MMR and adverse clinicopathological features on survival outcomes. Results of interaction analysis showed that the multiplication interaction effect between the number of lymph node harvest <12 and MMR protein status was significant on DFS of colon cancer patients ( hazard ratio=3.923, 95% confidence interval as 1.057-14.555, P<0.05). The additive interaction effects between age and MMR protein status, between high grade tumor and MMR protein status were significant on OS of colon cancer patients ( RERI=-0.033, -1.304, 95% confidence interval as -0.049 to -0.018, -2.462 to -0.146). Conclusions:There is an interaction between the MMR protein status and the adverse clinicopathological features (the number of lymph node harvest <12, high grade tumor) on prognosis of colon cancer patients of stage Ⅰ-Ⅲ. In patients of dMMR, the number of lymph node harvest <12 has a stronger predictive effect on poor prognosis. In patients of pMMR, the high grade tumor has a stronger predictive effect on poor prognosis.

Objective:To evaluate the clinical efficacy of ultrasound-guided radiofrequency ablation therapy for patients with papillary thyroid microcarcinoma (PTMC).Methods:A retrospective analysis was conducted on the clinical data of 80 PTMC patients admitted to Zhongshan Hospital of Traditional Chinese Medicine(Zhongshan Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Traditional Chinese Medicine) from January 2018 to September 2020. All cases were divided into two groups according to different treatment methods, with 40 cases in each group. The control group received PTMC radical surgery, while the observation group received ultrasound guided radiofrequency ablation. The situation of new tumors and cervical lymph node metastasis in both groups was observed through follow-up, the relative indexes of operation and safety of the two groups were compared.Results:There was no significant difference in the incidence of situation of new tumors and cervical lymph node metastasis between the two groups ( P>0.05). The operation time, intraoperative blood loss and hospital stay in the observation group were less than those in the control group: (20.23 ± 5.07) min vs. (54.51 ± 12.13) min, (1.68 ± 0.28) ml vs.(13.68 ± 2.31) ml, (1.82 ± 0.32) d vs. (5.43 ± 0.57) d, there were statistical differences ( P<0.05). The incidence of complications in the observation group was lower than that in the control group: 5.00%(2/40) vs. 25.00%(10/40), there was statistical difference ( P<0.05). Conclusions:Ultrasound guided radiofrequency ablation for the treatment of PTMC is safe and effective, minimally invasive in beauty, and can effectively reduce the incidence of complications.

Objective:To explore the outcome of sponge forceps assisted threading with Speedbridge technique for the treatment of acute closed Achilles tendon rupture.Methods:A retrospective case series study was conducted on 20 patients with acute closed Achilles tendon rupture treated in Zhengzhou Orthopedic Hospital from December 2019 to December 2021. There were 18 males and 2 females, with age range of 24-43 years [(29.5±7.6)years]. All patients were with unilateral injury, involving the left side in 13 patients and right side in 7. Examinations revealed a palpable defect in the Achilles tendon and positive Thompson test. A longitudinal incision was made at the medial edge of the ruptured tendon. Three nonabsorbable sutures were passed through the proximal stump with sponge forceps, bypassed the rupture site and fixed directly into the calcaneal bone. The disrupted tendon ends were aligned by the tendon-bundle technique using 4-0 absorbable sutures. The operation time and incision length were documented. The ankle joint range of motion (dorsiflexion/plantar flexion), American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and Achilles tendon total rupture score (ATRS) in the affected and healthy side were compared at 3, 6 and 12 months postoperatively. The wound healing and complications were observed.Results:All patients were followed up for 12-16 months [(13.2±2.5)months]. The operation time was 40-66 minutes [(52.0±10.3)minutes], with the incision length of 3-4 cm [(3.3±0.7)cm]. In the affected side at 3 and 6 months postoperatively, the ankle joint dorsiflexion [(5.6±1.5)°, (10.5±0.2)°] and plantar flexion [(28.4±3.2)°, (33.5±1.5)°] showed statistically significant difference compared with the healthy side (all P<0.05). The ankle joint dorsiflexion [(13.9±0.7)°] and plantar flexion [(38.3±4.4)°] in the affected side were not statistically different from that of the healthy side at 12 months postoperatively (all P>0.05). The AOFAS ankle-hindfoot score was (58.3±5.4)points, (84.9±7.1)points and (91.8±6.3)points at 3, 6 and 12 months postoperatively, showing a gradual rise (all P<0.05). The ATRS was (60.5±4.9)points, (85.5±9.0)points and (93.1±5.7)points at 3, 6 and 12 months postoperatively, showing a gradual rise (all P<0.05). All incisions were healed primarily. No patients had wound infection, nerve injury or re-rupture. Pain at the anchor insertion site occurred in 2 patients at 1 month after operation and relieved after active functional rehabilitation at 4 months after operation. Transient pain at the Achilles tendon insertion occurred in 1 patient at 6 months after operation, and relieved after 2 weeks of oral non-steroidal anti-inflammatory drugs treatment. Conclusion:For acute closed Achilles tendon rupture, sponge forceps assisted threading with Speedbridge technique can attain short operation time, small incision and good functional recovery, with few complications.

Preoperative sleep loss can amplify post-operative mechanical hyperalgesia. However, the underlying mechanisms are still largely unknown. In the current study, rats were randomly allocated to a control group and an acute sleep deprivation (ASD) group which experienced 6 h ASD before surgery. Then the variations in cerebral function and activity were investigated with multi-modal techniques, such as nuclear magnetic resonance, functional magnetic resonance imaging, c-Fos immunofluorescence, and electrophysiology. The results indicated that ASD induced hyperalgesia, and the metabolic kinetics were remarkably decreased in the striatum and midbrain. The functional connectivity (FC) between the nucleus accumbens (NAc, a subregion of the ventral striatum) and the ventrolateral periaqueductal gray (vLPAG) was significantly reduced, and the c-Fos expression in the NAc and the vLPAG was suppressed. Furthermore, the electrophysiological recordings demonstrated that both the neuronal activity in the NAc and the vLPAG, and the coherence of the NAc-vLPAG were suppressed in both resting and task states. This study showed that neuronal activity in the NAc and the vLPAG were weakened and the FC between the NAc and the vLPAG was also suppressed in rats with ASD-induced hyperalgesia. This study highlights the importance of preoperative sleep management for surgical patients.
Rats ; Animals ; Hyperalgesia/metabolism* ; Sleep Deprivation/metabolism* ; Rats, Sprague-Dawley ; Periaqueductal Gray/pathology* ; Proto-Oncogene Proteins c-fos/metabolism* ; Pain, Postoperative/pathology*

Objective     To verify the feasibility and accuracy of the "lung surface intersegmental constant proportion landmarks", developed by our center, in identifying intersegmental planes during pulmonary segmentectomy. Methods    We prospectively enrolled the patients who planned to receive thoracoscopic segmentectomy in West China Hospital of Sichuan University and The Third People's Hospital of Chengdu from September 2021 to October 2021. We took a relatively objective and feasible method, intravenous injection of indocyanine green, in identifying intersegmental planes as standard control. We intraoperatively judged the consistency between "lung surface intersegmental constant proportion landmarks" and intravenous injection of indocyanine green in identifying intersegmental planes. We discerned main landmarks of intersegmental plane by the constant proportion segment module, which was built based on the "lung surface intersegmental constant proportion landmarks", as well as distinguished the planes with discrepant fluorescence by peripheral intravenous indocyanine green injection. When the distance between the landmarks determined by the "ung surface intersegmental constant proportion landmarks" and the segmental boundaries displayed by indocyanine green  fluorescence staining was ≤1 cm, the landmarks were judged to be consistent with the planes with discrepant fluorescence. As long as one of the landmarks was judged to be consistent, the method was considered to be feasible and accurate. Results 聽 聽 A total of 21 patients who underwent thoracoscopic segmentectomy were enrolled, with 5 male and 16 female patients. The median age was 55 years, ranging from 34 to 76 years. A total of 11 patients received left-side surgery, while 10 patients received right-side surgery. In the operations of 21 pulmonary segmentectomies, at least one intersegmental landmark determined by the "lung surface intersegmental constant proportion landmarks" was consistent with the intersegmental plane determined by indocyanine green fluorescence staining in each patient. Conclusion 聽 聽The intersegmental landmarks determined by the "lung surface intersegmental constant proportion landmarks" are consistent with that determined by indocyanine green fluorescence staining. The method of "lung surface intersegmental constant proportion landmarks" is feasible and accurate in identifying intersegmental planes during pulmonary segmentectomy.

Objective To evaluate Evans lateral lengthening calcaneal osteotomy(E-LLCOT) in the treatment oftalocalcaneal coalitions (TCCs) with forefoot abduction deformity in the teenagers.Methods From February 2014 to August 2018,11 teenaged patients (14 feet) were treated at Department of Foot and Ankle Surgery,Zhengzhou Orthopaedics Hospital for TCCs with severe forefoot abduction deformity.They were 6 males (8 feet) and 5 females (6 feet),aged from 13 to 17 years (average,15 years).Their diseases involved bilateral feet in 3 cases and unilateral foot in 8,the left foot in 7 cases and the right in 7.All patients underwent TCCs resection followed by E-LLCOT.Their talonavicular coverage angles (TCA) and talar-second metatarsal angles (T2-MT) on the anteroposterior film and talar horizontal angles (TH) and talar-first metatarsal angles (T1-MT) on the lateral film were measured preoperatively and at the last follow-up.The foot functions were evaluated preoperatively and at the last follow-up using the ankle-hindfoot scores of American Orthopaedic Foot and Ankle Society (AOFAS) and visual analogue scale (VAS).Results All the 11 patients were followed up for 12 to 24 months (average,16.5 months).The mean preoperative TCA (22.3°,from 20° to 26°) was improved to 10.5° (from 8° to 13°) at the last follow-up;the mean T-2MT was improved from preoperative 17.6° (from 16° to 20°) to 6.5° (from 5° to 11°) at the last follow-up;the mean TH on the lateral view was improved from preoperative 35° (from 25° to 40°) to 17.5° (from 16° to 21°) at the last follow-up;the mean T-1MT was improved from preoperative 15.5° (from 10° to 22°) to 3.5° (from 2° to 6°) at the last follow-up;the mean AOFAS score was improved from 56.5 (from 50 to 62) preoperatively to 90.6 (from 75 to 95) at the last follow-up;the mean VAS score was improved from 6.0 (from 5 to 7) preoperatively to 2 (from 0 to 3) at the last follow-up.Conclusion For TCCs with severe forefoot abduction deformity in the geenagers,E-LLCOT after TCCs resection can effectively correct deformity,relieve pain and achieve significant functional and radiographic improvements.

Objective:To analyze the therapeutic effect of ankle arthrodesis with reverse proximal humerus internal locking system plating plus cannulated screwing via the transfibular approach in the treatment of end-stage ankle arthrosis.Methods:From April 2014 to January 2018, 19 patients (19 ankles) with end-stage ankle osteoarthritis were treated at Department of Foot and Ankle Surgery, Zhengzhou Orthopaedics Hospital by ankle arthrodesis with reverse PHILOS plating plus cannulated screwing via the transfibular approach for multi-plane fixation. They were 9 men and 10 women, aged from 36 to 65 years (average, 46.7 years). The disorder was further diagnosed as traumatic arthritis in 9 cases, as osteoarthritis in 6, as talar necrosis in 2 and as equinovarus in 2, and affected the left ankle in 11 cases and the right in 8. All cases belonged to stage Ⅲ osteoarthritis according to the Morrey-Wiedeman imaging classification. Their American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score, visual analogue scale (VAS), joint fusion and complications were observed preoperatively and at the last follow-up.Results:All the 19 patients were followed up for 18 to 62 months (average, 31.5 months). Their AOFAS scores were improved from preoperative 46.3±2.1 to 81.6±0.9 at the last follow-up, and their VAS was reduced from preoperative 7.1±0.2 to 2.0±0.2 at the last follow-up, showing significant differences ( P<0.001). All their ankle joints obtained bony fusion after 9 to 14 weeks (mean, 11.5 weeks). No implant loosening or breakage was observed. All the patients were satisfied with their operative results at the last follow-up. Dorsal lateral foot numbness was observed in one patient who became asymptomatic after neurotrophic medication for 3 months. Lateral distal wound dehiscence happened during removal of stitches at 2 weeks after operation in another patient but healed after debridement. The last follow-up found subtalar joint degeneration in 2 cases and talonavicular joint degeneration in one but no clinical symptoms in the 3. Conclusion:Ankle arthrodesis with reverse PHILOS plating plus cannulated screwing via the transfibular approach is a fine treatment for end-stage ankle arthrosis, because it leads to reliable fixation, short fusion time, alleviated pain and improved ankle function.

Objective To present the initial clinical experience of robot-assisted thoracoscopic esophagectomy for patients with esophageal cancer and to analyze the short-term outcomes of these patients. Methods Between February 2016 and December 2017, 148 patients with esophageal carcinoma underwent robotic esophagectomy and two-fields lymph node dissection. There were 126 males and 22 females at average age of 62.0±8.0 years. Demographic data, intraoperative characteristics and short-term surgical outcomes were collected and analyzed. Results 106 patients underwent McKeown esophagectomy and 42 patients underwent Ivor-Lewis esophagectomy. The mean operation time was 336.0±76.0 min, the mean intraoperative blood loss was 130.0±89.0 ml, the mean number of lymph nodes removed was 21.0±8.0 and the mean length of postoperative hospital-stay was 12.0±7.2 days. Postoperative complications included anastomotic fistula (n=8, 5.4%), pulmonary infection (n=13, 8.7%), hoarseness (n=23, 15.5%), tracheoesophageal fistula (n=1, 0.7%), chylothorax (n=4, 2.7%) and incision infection (n=2, 1.4%). There was no intra-operational massive hemorrhage or in-hospital mortality. Conclusion Both robot-assisted McKeown and Ivor-Lewis esophagectomy are safe and feasible with good early outcomes.