[Objective] To establish a cryopreservation protocol for small-volume (≤1 mL) red blood cells (RBCs) and to evaluate the reactivity and stability of blood group antigens after cryopreservation, so as to explore its potential application in immunohematology reference laboratories. [Methods] Small-volume RBCs were cryopreserved for 120 days, followed by thawing and deglycerolization to restore the RBC components. The quality of the RBCs was assessed. Serum antibodies were serially diluted and reacted with RBCs before and after cryopreservation, and agglutination scores were recorded to quantitatively evaluate the reactivity and stability of blood group antigens such as Rh, Duffy, Lewis, Kidd, M, and H. Flow cytometry was used to analyze the percentage and mean fluorescence intensity of ABO antigen expression on RBCs before and after cryopreservation to assess the usability of cryopreserved RBCs in flow immunophenotyping and blood group subtype studies. [Results] The hemolysis rate of thawed and deglycerolized RBCs was (0.27±0.10)%, with a supernatant free hemoglobin level of (0.52±0.14) g/L, and the RBC recovery rate was (69.12±7.91)%. The direct antiglobulin test (DAT) was negative for all thawed RBCs. There was no difference in the reactivity of blood group antigens before and after cryopreservation, and no difference in the percentage and mean fluorescence intensity of A and B antigen expression on RBCs before and after cryopreservation. [Conclusion] The small-volume RBC cryopreservation protocol can be applied to immunohematology analysis in reference laboratories and is expected to be widely used in blood group identification, antibody screening, identification, and blood group-related research.

Urticarial vasculitis is a skin disease with urticaria-like lesions and a histopathological pattern of leukocytoclastic vasculitis. It is considered a "hidden rash" in traditional Chinese medicine. Xuanfu is the portal that regulates qi, blood, fluid, and the ascending, descending, exiting, and entering of nutrition qi and defensive qi. Collaterals are the pathways for the circulation of qi and blood. The two accompany each other, connecting zang-fu organs, reaching the surfaces of the skin, hair, and external body, circulating qi and fluid, and moistening and protecting the skin. Based on the theory of Xuanfu-collateral, this study aimed to clarify the etiology, pathogenesis, and treatment method of urticarial vasculitis. External assault by wind and Xuanfu blockage are believed to be the initiating factors of this disease. The malnutrition of Xuanfu and collaterals and accumulated dampness-heat are important links in the occurrence and development of urticarial vasculitis. It spreads from Xuanfu to the collaterals, and blockage of the collaterals is the immanent trend of this disease. Clinically, by closely adhering to the core pathogenesis of blockage of Xuanfu-collateral, treatment method such as using wind medicinals to open Xuanfu with pungent and dispersing properties, using the method of supplement deficiency and removing the blockage, and using medicinals to promote blood circulation and remove blood stasis to unblock the blocked collaterals. The herbs are flexibly added or subtracted to unblock Xuanfu and collaterals, harmonize qi and blood, thus all symptoms can be relieved. We hope that this study will provide new ideas for the treatment of urticarial vasculitis with traditional Chinese medicine.

Drug administration via hollow microneedles (HMN) have the advantages of painlessness, avoidance of first-pass effect, capability of sustained infusion, and no need for professional personnel operation. In addition, HMN can also be applied in the fields of body fluid extraction and biosensors, showing broad application prospects. However, traditional manufacturing technologies cannot meet the demand for low-cost mass production of HMN, limiting its widespread application. This paper reviews the main manufacturing technologies used for HMN in recent years, which include photolithography and etching, laser etching, sputtering and electroplating, micro-molding, three-dimensional (3D) printing and drawing lithography. It further analyzes the characteristics and limitations of existing manufacturing technologies and points out that the combination of various manufacturing technologies can improve production efficiency to a certain extent. In addition, this paper looks forward to the future trends of HMN manufacturing technology and proposes possible directions for its development. In conclusion, it is expected that this review can provide new ideas and references for follow-up research.
Printing, Three-Dimensional ; Needles ; Humans ; Drug Delivery Systems/methods* ; Equipment Design ; Microinjections/methods*

Microbial communities such as those residing in the human gut are highly diverse and complex, and many with important implications for health and diseases. The effects and functions of these microbial communities are determined not only by their species compositions and diversities but also by the dynamic intra- and inter-cellular states at the transcriptional level. Powerful and scalable technologies capable of acquiring single-microbe-resolution RNA sequencing information in order to achieve a comprehensive understanding of complex microbial communities together with their hosts are therefore utterly needed. Here we report the development and utilization of a droplet-based smRNA-seq (single-microbe RNA sequencing) method capable of identifying large species varieties in human samples, which we name smRandom-seq2. Together with a triple-module computational pipeline designed for the bacteria and bacteriophage sequencing data by smRandom-seq2 in four human gut samples, we established a single-cell level bacterial transcriptional landscape of human gut microbiome, which included 29,742 single microbes and 329 unique species. Distinct adaptive response states among species in Prevotella and Roseburia genera and intrinsic adaptive strategy heterogeneity in Phascolarctobacterium succinatutens were uncovered. Additionally, we identified hundreds of novel host-phage transcriptional activity associations in the human gut microbiome. Our results indicated that smRandom-seq2 is a high-throughput and high-resolution smRNA-seq technique that is highly adaptable to complex microbial communities in real-world situations and promises new perspectives in the understanding of human microbiomes.
Humans ; Gastrointestinal Microbiome/genetics* ; Bacteriophages/physiology* ; High-Throughput Nucleotide Sequencing ; Sequence Analysis, RNA/methods* ; Bacteria/virology*

Objective To explore the automated identification and diameter measurement methods for inferior vena cava (IVC) based on clinical ultrasound images of IVC. Methods An automated identification and localization method based on topology and automatic tracking algorithm was proposed. Tracking algorithm was used for identifying and continuously locating to improve the efficiency and accuracy of measurement. Tests were conducted on 18 sets of ultrasound data collected from 18 patients in intensive care unit (ICU),with clinicians' measurements as the gold standard. Results The recognition accuracy of the automated method was 94.44％ (17/18),and the measurement error of IVC diameter was within the range of±1.96s (s was the standard deviation). The automated method could replace the manual method. Conclusion The proposed IVC automated identification and localization algorithm based on topology and automatic tracking algorithm has high recognition success rate and IVC diameter measurement accuracy. It can assist clinicians in identifying and locating IVC,so as to improve the accuracy of IVC measurement.

Objective:Glioblastoma(GBM)and brain metastases(BMs)are the two most common malignant brain tumors in adults.Magnetic resonance imaging(MRI)is a commonly used method for screening and evaluating the prognosis of brain tumors,but the specificity and sensitivity of conventional MRI sequences in differential diagnosis of GBM and BMs are limited.In recent years,deep neural network has shown great potential in the realization of diagnostic classification and the establishment of clinical decision support system.This study aims to apply the radiomics features extracted by deep learning techniques to explore the feasibility of accurate preoperative classification for newly diagnosed GBM and solitary brain metastases(SBMs),and to further explore the impact of multimodality data fusion on classification tasks. Methods:Standard protocol cranial MRI sequence data from 135 newly diagnosed GBM patients and 73 patients with SBMs confirmed by histopathologic or clinical diagnosis were retrospectively analyzed.First,structural T1-weight,T1C-weight,and T2-weight were selected as 3 inputs to the entire model,regions of interest(ROIs)were manually delineated on the registered three modal MR images,and multimodality radiomics features were obtained,dimensions were reduced using a random forest(RF)-based feature selection method,and the importance of each feature was further analyzed.Secondly,we used the method of contrast disentangled to find the shared features and complementary features between different modal features.Finally,the response of each sample to GBM and SBMs was predicted by fusing 2 features from different modalities. Results:The radiomics features using machine learning and the multi-modal fusion method had a good discriminatory ability for GBM and SBMs.Furthermore,compared with single-modal data,the multimodal fusion models using machine learning algorithms such as support vector machine(SVM),Logistic regression,RF,adaptive boosting(AdaBoost),and gradient boosting decision tree(GBDT)achieved significant improvements,with area under the curve(AUC)values of 0.974,0.978,0.943,0.938,and 0.947,respectively;our comparative disentangled multi-modal MR fusion method performs well,and the results of AUC,accuracy(ACC),sensitivity(SEN)and specificity(SPE)in the test set were 0.985,0.984,0.900,and 0.990,respectively.Compared with other multi-modal fusion methods,AUC,ACC,and SEN in this study all achieved the best performance.In the ablation experiment to verify the effects of each module component in this study,AUC,ACC,and SEN increased by 1.6%,10.9%and 15.0%,respectively after 3 loss functions were used simultaneously. Conclusion:A deep learning-based contrast disentangled multi-modal MR radiomics feature fusion technique helps to improve GBM and SBMs classification accuracy.

Objective:This study investigates the influence of the apolipoprotein E ε4(APOE ε4)genotype on the relationship between glucose-lipid metabolism disorders and diabetes-related cognitive impairment(DCI).Methods:A case-control study was conducted involving 891 patients with type 2 diabetes mellitus(T2DM)with a mean age of(62.1±13.8)years, all of whom underwent elective surgery at the First Hospital of Shanxi Medical University between January 2017 and December 2022.Among these participants, 229 were diagnosed with DCI(case group), while 662 were cognitively normal(control group).Routine clinical information was collected, and peripheral venous blood samples were analyzed for glycated hemoglobin(HbA1c)and blood lipid levels.The single nucleotide polymorphisms rs429358 and rs7412 were analyzed to determine the presence of the APOE ε4 genotype.Stepwise Logistic regression was employed to identify independent risk factors for DCI, and subgroup analyses were performed to evaluate the effect of the APOE ε4 genotype on the relationship between HbA1c and blood lipid levels in relation to DCI risk. Results:Among all patients, female gender( OR=1.915, 95% CI: 1.393-2.631, P<0.001), longer duration of T2DM( OR=1.169, 95% CI: 1.087-1.257, P<0.001), elevated triglycerides( OR=1.161, 95% CI: 1.041-1.294, P=0.007), and being an APOE ε4 carrier( OR=1.638, 95% CI: 1.115-2.405, P=0.012)were identified as independent risk factors for developing DCI.High levels of low-density lipoprotein(LDL)were found to be independently associated with an increased risk of DCI specifically in APOE ε4 carriers( OR=1.408, 95% CI: 1.060-1.870, P=0.018), but not in non-APOE ε4 carriers( P>0.05).In contrast, elevated HbA1c was independently associated with a higher risk of DCI in non-APOE ε4 carriers( OR=1.220, 95% CI: 1.040-1.430, P=0.014), but not in APOE ε4 carriers( P>0.05).Additionally, elevated triglycerides were independently linked to an increased risk of DCI across the entire sample and within each APOE ε4 genotype subgroup. Conclusions:The APOE genotype plays a significant role in modulating the relationship between dyslipidemia and the risk of developing DCI.This highlights the critical importance of lipid metabolism disorders and APOE risk genes in both the development and progression of DCI.These findings offer valuable insights for future clinical and mechanistic studies focused on DCI.

Objective:To explore the feasibility of deep learning-based restoration of obscured thyroid ultrasound images.Methods:A total of 358 images of thyroid nodules were retropectively collected from January 2020 to October 2021 at Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, and the images were randomly masked and restored using DeepFillv2. The difference in grey values between the images before and after restoration was compared, and 6 sonographers (2 chief physicians, 2 attending physicians, 2 residents) were invited to compare the rate of correctness of judgement and detection of image discrepancies. The ultrasound features of thyroid nodules (solid composition, microcalcifications, markedly hypoechoic, ill-defined or irregular margins, or extrathyroidal extensions, vertical orientation and comet-tail artifact) were extracted according to the Chinese Thyroid Imaging Reporting and Data System (C-TIRADS). The consistency of ultrasound features of thyroid nodules before and after restoration were compared.Results:The mean squared error of the images before and after restoration ranged from 0.274 to 0.522, and there were significant differences in the rate of correctness of judgement and detection of image discrepancies between physicians of different groups(all P<0.001). The overall accuracy rate was 51.95%, the overall detection rate was 1.79%, there were significant differences also within the chief physicians and resident groups (all P<0.001). The agreement rate of all ultrasound features of the nodules before and after image restoration was higher than 70%, over 90% agreement rate for features such as solid composition and comet-tail artifact. Conclusions:The algorithm can effectively repair obscured thyroid ultrasound images while preserving image features, which is expected to expand the deep learning image database, and promote the development of deep learning in the field of ultrasound images.

Objective:To investigate the clinical application of multimodal low-opioid combined with saphenous nerve block analgesia in elderly patients undergoing knee arthroplasty.Methods:This study is a randomized controlled study.A total of 60 elderly patients who underwent elective knee arthroplasty in the Department of Orthopedics, the First Hospital of Shanxi Medical University from January 2021 to December 2021 were selected and divided into 2 groups by numerical randomization: mode low opioid analgesia regimen group(observation group)and traditional analgesia regimen group(control group), 30 cases in each group.Observation group: (1)Preemptive analgesia: Oral celecoxib 200 mg, qd.from 3 days before surgery, the mini-mental state examination(MMSE)score was used to evaluate the cognitive function; (2)Intraoperative analgesia: After the prosthesis was installed, choose to inject analgesics around the knee joint(ropivacaine 200 mg, morphine 5 mg, epinephrine 0.25 mg, dexamethasone 5 mg/100 ml normal saline, also known as "cocktail" solution); (3)Postoperative analgesia: After the operation, continuous saphenous nerve block(0.2% ropivacaine, 2 ml/h)was performed under ultrasound guidance, and the dose of nerve block was adjusted according to the degree of rehabilitation training.Control group: no special treatment before and during the operation, traditional postoperative patient-controlled intravenous analgesia(PCIA)was used after the operation, the formula WAs as follows: sufentanil(2 μg/kg)+ flurbiprofen axetil(200-300 mg)+ Tropisetron(5-10 mg). The numerical rating scale(NRS)scores were observed and recorded 1 day before surgery, 6, 12, 24, and 48 hours after surgery; the time of the first active straight leg raising after surgery, the time of landing; the range of motion(ROM)of the knee joint 3, 7, and 14 days after surgery; the American Hospital for Special Surgery Knee Score(HSS)at 14 days, 1, 3, and 6 months after surgery; serum substance P(SP)and interleukin-6(IL-6)concentrations 1 day before surgery and 48 hours after surgery, and adverse events were recorded.Results:Compared with the control group, the patients in the observation group had lower NRS scores at 6, 12, 24, and 48 hours after the operation( t=27.705, 27.532, 21.739, 25.780, all P<0.05); the first active straight leg raising time and the time of landing earlier after the operation, and the knee joint at 3, 7, and 14 days after the operation, the range of motion(ROM)was better( t=35.496, 43.716, 3.766, 5.216, 6.009, all P<0.05). And the American hospital for special surgery knee score(HSS)was higher at 14 days, 1 month, and 3 months after surgery( t=19.247, 32.337, 22.651, all P<0.05), but there was no significant difference at 6 months after surgery.Simultaneously, the serum SP and IL-6 concentrations in the observation group 48 h after the operation were (431.0±11.3)ng/L and(11.9±2.7)ng/L, respectively.Compared with the control group(442.5±15.6)ng/L, (14.4±2.9)ng/L( t=5.362, 4.144, both P<0.05). Compared with the control group, which were lower than those in the incidence of postoperative nausea and vomiting was lower, the length of hospital stay was shorter in the observation group( χ2=4.630, t=3.311, P=0.031, 0.002), and the other indicators had no statistical differences(all P>0.05). Conclusions:Multimodal low-opioid combined with saphenous nerve block analgesia can significantly reduce perioperative pain in elderly patients undergoing knee arthroplasty, improve early postoperative mobility, and speed up postoperative functional recovery.

Objective:To explore the effect of spontaneous breathing during induction of general anesthesia on atelectasis in patients undergoing laparoscopic resection of gastrointestinal tumors.Methods:A total of 60 patients aged 18-60 years scheduled for laparoscopic resection of gastrointestinal tumors under general anesthesia in the First Hospital of Shanxi Medical University from October 2021 to August 2022 were selected. The body mass index was 18.5-28.0 kg/m 2 and the American Society of Anesthesiology grade wasⅠ-Ⅱ. All patients were divided into the spontaneous breathing group (group S, 30 cases) and the controlled breathing group (group C, 30 cases) according to the random number table method. Patients in group S received 0.2-0.3 mg/kg etomidate (pumping at the speed of 200 ml/h) and 2 μg/kg remifentanil (slowly injected more than 30 s) for anesthesia induction; patients in group C received 0.2-0.3 mg/kg etomidate and 2 μg/kg remifentanil (slowly injected more than 30 s) and 0.2 mg/kg cisatracurium. After bispectral index (BIS) decreased to 80, the patients had no response to the language stimulation; and then the mask was used to closely fit the face and maintain spontaneous breathing in group S; patients in group C received manual positive pressure ventilation. Atelectasis scores were collected immediately after endotracheal intubation (T 1) and 15 min after transferring to the recovery room (T 3), and oxygenation index (OI) was collected 5 min after endotracheal intubation (T 2) and at T 3. The postoperative pulmonary complication (PPC) on the 3rd day after the operation was recorded. Results:A total of 56 patients were finally enrolled, 27 cases in group S and 29 cases in groups C. Compared with group C, the atelectasis score of group S at T 1 and T 3 decreased [T 1: (2.4±0.8) scores vs. (4.2±0.7) scores, t = -9.12, P < 0.001; T 3: (8.2±1.8) scores vs. (10.5±1.6) scores, t = -4.96, P < 0.001]. The OI increased at T 2 and T 3 in group S [T 2: (334±11) mmHg (1 mmHg = 0.133 kPa) vs. (323±13) mmHg, t = 3.45, P = 0.001; T 3: (362±23) mmHg vs. (347±25) mmHg, t = 2.31, P = 0.025]. The incidence of PPC was 20.7% (6/29) and 18.5% (5/27), respectively in group C and group S on the 3rd day after the operation, and the difference was statistically significant ( χ2 = 0.04, P = 0.838). Conclusions:Maintaining spontaneous breathing during induction of general anesthesia can reduce atelectasis caused by general anesthesia and improve oxygenation for patients undergoing laparoscopic resection of gastrointestinal tumors.