Objective: To optimize the technology and process for recovering immune cells from discarded leukocyte filter through reverse flushing, and to investigate the effects of different reverse flushing solutions and physical conditions (temperature, soaking, and ultrasound) on the cell recovery rate. Additionally, to evaluate the feasibility and quality of isolating and culturing NK cells from the recovered cells. Methods: Normal saline, PBS, and PBS-EDTA were used as reverse flushing solutions to backwash the filter, and the white blood cell recovery rates were compared. The reverse flushing efficiency was optimized by adjusting the temperature of the reverse flushing solution, filter soaking treatment and ultrasonic treatment. The Ficoll separation method was employed to isolate peripheral blood mononuclear cells (PBMCs) from the elution solution and conduct in vitro expansion and culture. Lymphocyte subsets were identified and quantified using flow cytometry. Results: The physiological saline served as the backflush fluid was the most stable. The reverse temperature had a significant impact on the recovery rate of white blood cell subgroups. The recovery rates of lymphocytes and monocytes at 37℃ were significantly higher than those at 4℃ [(35.56±5.55)% vs (22.88±0.31)%, (32.42±4.82)% vs (8.00±1.6)%, t=5.581 and 5.758, respectively, both P<0.001)], and at 25℃ [(35.56±5.55)% vs (22.42±6.13)%, t=5.904, P<0.001; (32.4±4.82)% vs (21.11±1.23)%, t=2.629, P=0.030)]. The recovery rates of white blood cell subgroups after 37℃ immersion were higher than those after direct reverse flow at 37℃, but the difference was not statistically significant (P>0.05). The recovery rate of cells after ultrasonic immersion at 25℃ was further increased compared to that after immersion at 25℃, but the difference was not statistically significant (P>0.05). The purity of the NK cells obtained through sorting reached (94.2±4.5)%, and the viability was (94.8±0.9)%. After 15 days of culture, the cell count was expanded approximately 450 times, yielding more than 10 /mL highly active NK cells. Conclusion: By optimizing the backflushing process, immune cells can be efficiently recovered from discarded filters. The 37℃ backflow liquid has the best backflow effect; soaking and ultrasonic treatment can be used as auxiliary methods to improve recovery rates. The NK cells from this source have high purity, good activity, and strong expansion ability, and can be used as a reliable cell source for immunotherapy research, and in line with the circular economy concept of resource reuse.

Objective To observe the clinical efficacy of Lingnan fire needling in the treatment of plaque psoriasis.Methods A total of 60 cases of patients with plaque psoriasis admitted to Bao'an Hospital of Chinese Medicine Affiliated to Guangzhou University of Chinese Medicine from August 2021 to January 2023 were selected,and the patients were randomly divided into the observation group and the control group according to the random number table method,with 30 cases in each group.The observation group was given Lingnan fire needling for treatment,and the control group was given topical application of Calcipotriol Ointment.The course of treatment covered eight weeks continuously.After eight weeks of treatment,the clinical efficacy of the two groups was evaluated,and the changes of Psoriasis Area and Severity Index(PASI)scores and Dermatology Life Quality Index(DLQI)scores,as well as the diameter of glomus under dermoscopy of the patients in the two groups were observed before the treatment and after 2,4,6,and 8 weeks of treatment.Changes in Generalized Anxiety Disorder Scale(GAD-7)scores before treatment and after 2,4,6,and 8 weeks of treatment were compared between the two groups.Results(1)After 2,4,6 and 8 weeks of treatment,the scores of PASI,DLQI and GAD-7 of patients in the two groups were improved significantly(P<0.05),and the improvement in the observation group was significantly superior to that in the control group,the difference being statistically significant(P<0.05).(2)The cured and markedly rate of the observation group was 86.67%(26/30);and that of the control group was 66.67%(20/30).The efficacy of the observation group was superior to that of the control group,the difference being statistically significant(P<0.05).(3)After 2,4,6,and 8 weeks of treatment,the diameter of glomus under dermoscopy of the patients in the two groups were improved significantly(P<0.05),and the improvement in the observation group was significantly superior to that in the control group,the difference being statistically significant(P<0.05).Conclusion Lingnan fire needling therapy for plaque psoriasis can effectively improve the clinical symptoms of the patients,relieve the anxiety symptoms induced by skin lesions and itching,so as to improve their quality of life.

Objective:To explore the value of the Nomogram model established by CT radiomics combined with clinical indicators for prediction of the severity of early acute pancreatitis (AP).Methods:From January 2016 to March 2023, the AP patients in the Second Affiliated Hospital of Soochow University were retrospectively collected. According to the revised Atlanta classification and definition of acute pancreatitis in 2012, all patients were divided into the severe group and the non-severe group. All patients were first diagnosed, and abdominal CT plain scan and enhanced scan were completed within 1 week. Patients were randomly (random number) divided into training and validation groups at a ratio of 7:3. The pancreatic parenchyma was delineated as the region of interest on each phase CT images, and the radiomics features were extracted by python software. LASSO regression and 10-fold cross-validation were used to reduce the dimension and select the optimal features to establish the radiomics signature. Multivariate Logistic regression was used to select the independent predictors of severe acute pancreatitis (SAP), and a clinical model was established. A Nomogram model was established by combining CT radiomics signature and clinical independent predictors. Receiver operating characteristic (ROC) curve and decision curve analysis (DCA) were used to evaluate the predictive efficacy of each model.Results:Total of 205 AP patients were included (59 cases in severe group, 146 cases in non-severe group). 3, 5, 5 and 5 optimal radiomics features were selected from the plain CT scan, arterial phase, venous phase and delayed phase images of all patients, and the radiomics models were established. Among them, the arterial phase radiomics model had relatively better performance in predicting SAP, with an area under curve (AUC) of 0.937 in the training group and 0.913 in the validation group. Multivariate Logistic regression showed that C-reactive protein (CRP) and lactate dehydrogenase (LDH) were independent predictors of SAP, and they were used to establish a clinical model. The AUC in the training and validation groups were 0.879 and 0.889, respectively. The Nomogram model based on arterial phase CT radiomics signature, CRP and LDH was established, and the AUC was 0.956 and 0.947 in the training group and validation group, respectively. DCA showed that the net benefit of Nomogram model was higher than that of clinical model or radiomics model alone.Conclusions:The Nomogram model established by CT radiomics combined with clinical indicators has high application value for early prediction of the severity of AP, which is conducive to the formulation of clinical treatment plans and prognosis evaluation.

Purpose To verify the safety and effectiveness of the Insight NM/CT Pro SPECT/CT Imaging System in clinical applications is verified through clinical validation of the acceptable rate of clinical image quality,commonly use features of the device,ease of use of the machine,satisfaction with the stability of the entire machine and safety evaluation.Materials and Methods A total of 79 subjects who voluntarily participated in the trial from July to October 2022 in the Affiliated Hospital of Inner Mongolia Medical University,were selected to undergo endocrine,circulatory,skeletal,respiratory and urological imaging for multiple systems using the Insight NM/CT Pro SPECT/CT,respectively.The quality of the images,the machine operation and safety were evaluated.Results The image quality of all 79 patients were evaluated.Of 36 cases,there were 35 cases of planar imaging were acceptable;Of 43 cases,there were 42 cases of tomographic imaging were acceptable,with an acceptability rate of 97.22%and 97.67%,respectively,and a lower limit of 95%CI of 85.47%and 87.71%,respectively,which were both higher than those of the target value(83.00%).When the operators performed imaging operations on 79 patients,the common functions of the equipment,the ease of use of the equipment and the stability of the whole machine were evaluated as a satisfaction rate of 100%,with no adverse events.Conclusion Insight NM/CT Pro SPECT/CT effectiveness fully meets the requirements of clinical applications,with clear images of tissue structure,full range of commonly used equipment features,stable system,and superior safety and reliability.

Objective:To establish and validate a nomogram model for early prediction of the risk of acute pancreatitis (AP) progressing to severe acute pancreatitis (SAP).Methods:CT signs and clinical laboratory parameters of 361 AP patients admitted to our Hospital from January 2016 to July 2022 were retrospectively collected. There were 221 males (61.2%) and 140 females (38.8%). According to the Atlantic score, all patients were divided into the SAP group (64 cases) and the non-SAP (NSAP) group (297 cases). Univariate analysis was used to screen out variables with statistically significant differences. Multivariate Logistic regression analysis was used to screen out the independent risk factors of SAP, and finally a nomogram prediction model was established. Receiver operating characteristic (ROC) curve, calibration curve and decision curve (DCA) were used to evaluate the predictive efficacy, accuracy and clinical practicability of the model, and Bootstrap method was used to verify the model internally.Results:Univariate analysis and multivariate Logistic regression analysis showed that pleural effusion ( OR=7.353, 95% CI: 3.344-16.170), posterior pararenal space (PPS) involvement ( OR=3.149, 95% CI: 1.314-7.527), serum creatinine concentration (Cr) ( OR=1.027, 95% CI: 1.017-1.038) and serum calcium concentration (Ca 2+) ( OR=0.038, 95% CI: 0.009-0.166) were independent risk factors for SAP ( P<0.05). A Nomogram model was established based on these four factors. The area under the ROC curve (AUC) of this model was 0.905 (95% CI: 0.869-0.933), indicating high predictive efficiency. Internal verification showed that the model had good accuracy in predicting SAP, and C-index was 0.90. DCA analysis showed that the model had high clinical practicability. Conclusions:The Nomogram model combining pleural effusion, PPS involvement, Cr and Ca 2+ had a good effect on early prediction of SAP, which could provide a new reference tool for clinical diagnosis and treatment.

Objective:To investigate the clinicopathological features, treatment and prognosis of neuroendocrine carcinoma of the breast.Methods:Clinical data of 26 patients with neuroendocrine carcinoma of the breast admitted to the Northern Jiangsu People's Hospital from July 2013 to Mar 2021 were analyzed.Results:All 26 cases were female, the average aged of (62.81±11.95) years, the first clinical manifestations were painless breast masses, the average size being of (23.34±9.47) mm. At the time of diagnosis, regional lymph node metastasis was found in 4 cases, 1 case developed distant metastasis. Most patients' were on stage Ⅱ by TNM staging, molecular typing was Luminal A, and invasive mammary carcinoma with neuroendocrine differentiation was most common, with positive rates of ER and PR of 96%, the positive rate of CgA and Syn were 69% and 100%, and there was not positive expression of HER2. All patients received surgical treatment, 25 patients underwent postoperative adjuvant therapy. Twenty-five patients were followed up for a median follow-up time of 39.50 months. During the follow-up, 3 cases developed distant metastasis, 1 case died, the mean survival time was (40.81±26.90) months, there was ao satistically significant difference compared with invasive mammary carcinoma ( t=1.291, P=0.209). The mean disease free interval is (39.96±27.58) months. The overall survival and disease free survival at 1, 2 and 5 years are 100%, 100% and 87%, respectively. Conclusions:Neuroendocrine carcinoma of the breast occurs more frequently in elderly women, often with large tumor size, low rate of regional lymph node and distant metastasis, moderate histological grade, early clinical stage, and the molecular typing is mostly Luminal A.The overall prognosis is fair.

Objective:To investigate the effects of hydrogen rich-saline (HRS) on intestinal mucosal barrier in rat with intestinal ischemia/reperfusion injury (IIRI).Methods:Twenty-four healthy male Sprague-Dawley rats, aged 8 weeks, were randomly divided into 3 groups (8 in each group) by random number table method: sham group, model group and HRS group.Rats in HRS group were intraperitoneally injected with HRS (10 mL/kg) at 30 min of ischemia, and the same amount of normal saline was intraperitoneally injected in model group.After 45 min of ischemia and 6 h of reperfusion, rats were sacrificed.Serum and ileum were collected for further detection.Tumor necrosis factor alpha (TNF-α), interleukin (IL)- 1β and IL-17A expression levels in serum were detected by conducting enzyme-linked immunosorbent assay (ELISA). The localization expressions of tight junction protein Occludin was detected by immunohistochemical staining (IHC), while the localization expression of tight junction protein zonula occluden-1 (ZO-1) were detected by immunofluorescence staining (IF). The protein expression of Occludin, ZO-1, and Lysozyme were detected by performing Western blot.The mRNA expression of Lysozyme and α-defensin were detected by real-time PCR (qPCR).Results:ELISA results proved that the levels of serum TNF-α and IL-1β in HRS group rats were significantly lower than those in model group [(62.02±29.97) ng/L vs.(113.40±44.58) ng/L, (21.68±0.35) ng/L vs.(28.29±3.49) ng/L], while the level of IL-17A increased [(28.18±5.28) ng/L vs. (15.10±3.60) ng/L] (all P<0.05). IHC staining: compared with model group, the expression of Occludin in HRS group was uniform and continuous, and the staining was darker.IF results: compared with model group, the fluorescence signal intensity of ZO-1 in HRS group rats significantly increased, and the distribution was clear and continuous.Wes-tern blot results: compared with model group, the expression levels of Occludin and ZO-1 proteins in HRS group rats remarkably increased (0.79±0.06 vs. 0.54±0.04, 0.91±0.11 vs. 0.51±0.13), while Lysozyme protein decreased (1.50±0.40 vs. 2.99±0.80) (all P<0.05). qPCR results revealed that the expression level of Lysozyme mRNA in HRS group rats was lower than that in model group (1.64±0.33 vs. 2.20±0.40), while α-defensin mRNA obviously increased (0.82±0.19 vs. 0.47±0.13) (all P<0.01). Conclusions:HRS protects intestinal mucosal barrier by inhibiting the expression of tight junctions and the secretion of antimicrobial peptides in rat suffering from IIRI.

Objective:To compare modified load-sharing rip-stop (mLSRS) technique with suture bridge (SB) for repair of massive rotator cuff tears.Methods:A retrospective analysis was conducted of the 68 patients who had been treated for massive rotator cuff tears from January 2017 to June 2019 at Department of Orthopedics, Dongyang People's Hospital. They were 26 males and 42 females, aged (63.4±1.0) years (from 45 to 84 years), with 56 right and 12 left sides affected. Of them, 32 were treated by mLSRS and 36 by double-line SB. The 2 groups were compared in terms of visual analog scale (VAS), Constant-Murley scoring, University of California Los Angeles (UCLA) scoring and American Shoulder and Elbow Surgeons (ASES) scoring at the final follow-ups, and Sugaya ultrasonic evaluation of the repair integrity at postoperative 12 months.Results:There was no statistically significant difference in preoperative general data between the 2 groups, showing they were comparable ( P>0.05). Operations were successful in all patients who were followed up for 13 to 44 months (average, 19.7 months). No patient suffered from such complications as joint infection or anchor withdrawal. At the final follow-up, the mLSRS group scored respectively 0.9±0.6, 85.3±4.8, 33.1±1.0 and 86.4±5.0 in VAS, Constant-Murley, UCLA and ASES scores, significantly improved than their preoperative values (7.2±0.8, 47.0±3.1, 15.8±3.0 and 48.5±4.5) ( P<0.05); the SB group scored respectively 1.1±0.6, 86.6±3.4, 33.2±1.1 and 86.9±4.6 in the above indicators, also significantly improved than their preoperative values (7.3±0.7, 46.5±4.7, 14.4±2.7 and 48.8±4.3) ( P<0.05); there were no significant differences between the 2 groups in preoperative or postoperative values ( P> 0.05). The Sugaya ultrasonic evaluation showed no significant difference in rotator cuff healing between the 2 groups at postoperative 12 months ( P>0.05); according to the positions of re-tears, the SB group had significantly more type Ⅱ re-tears (20%, 2/10) than the mLSRS group (0, 0/4) ( P<0.05). Conclusion:Although both mLSRS and SB techniques can achieve satisfactory and comparable clinical outcomes in arthroscopic repair of massive rotator cuff tears, mLSRS may lead to fewer postoperative type Ⅱ re-tears than SB.

Objective To evaluate the clinical characteristics and the clinical outcomes of perianal abscess (PA) in neonates.Methods A retrospective review was performed on the collected data of 185 patients of PA in neonates prospectively admitted to Binzhou Medical University Hospital from January 2008 to December 2015.Patients were divided into 2 groups on the parents' intention:nonsurgical treatment and surgical treatment,the standard surgical treatment for PA was incision and drainage with the use of packing.The standard surgical treatment for PA was surgical incision drainage of lower abscess under local anesthesia by the use of filling tamponade iodoform gauze,while the patients receiving conservative treatment took hip bath perianally with topical 1 ∶ 5 000 potassium permanganate,besmearing erythromycin eye ointment outside locally.Incision-thread-drawing procedure was recommended in fistula-in-ano (FIA) after 6 months.Antibiotics were administered in all patients in the early days.The clinical data of age,gender,accompanying diseases,abscess amount and location,treatment approach,healing time and recurrence rates were analyzed with statistical method.Results All patients were boys,time of visiting hospital was 1-25 day,the average time 7.5 days;60 cases (32.4％)had neonatal diarrhea,45 cases (24.3％)had neonatal jaundice,but no patients had severe fever.A single skin lesion was present in 145 patients (78.4％),2 lesions in 30 patients (16.2％),and 10 patients had 3 lesions (5.4％).The most commonly affected sites were at 9 o'clock clockwise direction with 115 (62.2％)lesions on lithotomy position,followed by 3 o'clock clockwise direction with 65(35.1％) lesions by 1 o'clock clockwise direction with 3 (1.6％) lesions and 6 o'clock clockwise direction with 2 (1.1％) lesions.Bacteria cultures were obtained from 123 patients (90.4％,123/136 cases) of surgical treatment and 35 patients (71.4％,35/49 cases) of nonsurgical treatment obtained the results of bacteria culture.The average healing time was (21 ±2) days (10-60 days) in the surgical treatment group,and (36 ± 3) days (9-90 days) in the nonsurgical treatment group,7 out of 136(5.1％) patients had a recurrence with surgical treatment,incision drainage was performed again with the use of packing,and FIA was not found,10 out of 49 (20.4％) patients had a recurrence with nonsurgical treatment group,and 6 out of 49 (12.2％) were spontaneously resolves within the first year of life,4 out of 49 (8.1％) developed into FIA,incision-thread-drawing procedure was performed after 6 months.The significant difference was observed between and nonsurgical treatment and surgical treatment in healing time (t =-6.707,P =0.000),recurrence (x2 =11.347,P =0.001) and FIA formation rate (x2 =10.054,P=0.002).Conclusions PA is an entity in neonates.Incision and drainage of PA is an effective and safe therapy in the early days.Surgery for PA may result in low recurrence rates,a low rate of evolution toward FIA,and a short healing time,which should be considered as the primary treatment.The key procedure is to keep the drainage unobstructed by the use of filling gauze drainage to prevent crissum abscess recurrence.Postoperative care with antibiotics is effective to shorten hospital stays.

Objective To investigate the expression of inositol requiting enzyme1 α (IRE1 α) and tumor necrosis factor receptor-associated factor 2 (TRAF2) and its significance through establishing models of intestinal ischemia reperfusion injury (IIRI) in rats.Methods According to the random number table,50 male SD rats were randomly divided into 2 groups:sham operation group (n =10) and ischemia reperfusion (I/R) group (n =40).Sham group animals underwent laparotomy.I/R group rats were subjected to occlusion of the superior mesenteric artery for 30 min;then the blood flow was restored.I/R group animals were divided into 4 subgroups:2 h,6 h,12 h,24 h according to the time of reperfusion.Eight rats were examined based on the number of live rats in each subgroup.The HE staining pathological changes in intestinal samples were observed by the light microscope.The small intestinal epithelial cell apoptosis index (AI) was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL).The expression levels of intestinal tissues tumor necrosis factor α (TNF-α) and plasma intestinal fatty acid-binding protein (Ⅰ-FABP) were detected by ELISA tests.Situ end labeling method was used to detect intestinal cell AI.Western blot was applied to investigate the expression of endoplasmic reticulum stress(ERS) proteins IRE1α,phosphorylation IREIα (p-IRE1 α) and TRAF2 in all group rats intestinal tissues.Results (1)The pathological changes showed that the intestinal injury of I/R groups was more severe than that of sham group,especially at 6 h.(2) Compared with sham group,the expression levels of TNF-α [sham group (16.41 ± 4.44)ng/ L,2 h group:(79.71 ± 8.20) ng/L,6 h group:(131.70 ± 11.59) ng/L,12 h group:(94.23 ±7.66) ng/L,24 h group:(69.78 ± 9.58) ng/L],AI[sham group:(3.93 ±0.77)％,2 h group:(16.24 ± 1.97)％,6 h group:(42.19 ±2.40)％,12 h group:(37.79 ± 2.34)％,24 h goup:(10.38 ±1.46)％] and plasma Ⅰ-FABP [sham group:(0.65 ±0.10) × 103 ng/L,2 h group:(1.47 ±0.10) ×103 ng/L,6 h group:(2.36 ±0.17) ×103 ng/L,12 h group:(37.79 ±2.34) ×103 ng/L,24 group:(l.41 ±0.09) × 103 ng/L] were higher (F =231.462,149.032,162.491,all P ＜ 0.01).(3) The expression of TRAF-2 protein and p-IRE1 α/IRE1 α could be up-regulated after IIRI (F =40.473,59.59,P ＜ 0.01).The expression of these proteins was up-regulated 2 h after reperfusion,peaking at 6-12 h reperfusion,and then decreased at 24 h,and the variation tendencies of all groups were the same.Conclusions IIRI could induce ERS,activate IRE1 α and up-regulate TRAF2.IRE1α/TRAF2 mediating ERS might be involved in regulating the cell inflammation,apoptosis and increasing intestinal permeability after IIRI.