[Objective] To explore the feasibility of using whole blood as a source of NK cells for allogeneic CAR NK cell therapy and activated NK cell reinfusion therapy, and initially construct a technical system for the separation and purification of NK cells from whole blood. [Methods] All peripheral blood mononuclear cells (PBMCs) were enriched from 400 mL of whole blood by manual separation and machine separation, respectively. The erythrocyte loss rate, PBMCs number, NK cell purity of the two methods were compared. NK cells were sorted from PBMCs by three separation and enrichment methods as immunomagnetic bead negative selection method, platelet lysate culture expansion and PERCOLL density gradient separation method, and the purity and yield of NK cells, the activity of NK cells and the tumor-killing ability of the three separation and enrichment methods were compared. [Results] The proportion of NK cells in the lymphocyte population was higher in the manual separation method than in the machine separation method[(13.16±5.16)% vs (8.56±3.92)%, P<0.05]; the number PBMCs was lower in the manual separation method than in the machine separation method[(4.09±1.80)×10⁸vs (6.49±2.16)×10⁸, P<0.05], and there was no difference in the red blood cell loss between the two methods (P>0.05). The purity of NK cells isolated and enriched from PBMCs by manual separation method using immunomagnetic was (96.77±2.31)%; the yield was (56.27±10.47)%; the inhibition of tumor proliferation was (38.67±14.05)%; and the tumor killing rate was (19.90±8.05)%. The purity of NK cells isolated and enriched from PBMCs by manual separation method using platelet lysis culture expansion method was the highest at day 7, which was (54.84±15.80)%; the cell expansion multiple could reach 16.92±6.28 at day 7; the in vitro tumor killing rate of NK cells was (15.83±5.5)%; the tumor inhibition rate was (44.33±13.5)%; and there was no difference in the toxicity and activity of NK cells between the two methods (P>0.05). The purity of NK cells isolated and enriched by PERCOLL density gradient separation method was (15.83±5.82)%, and the yield was (14±6.25)%, which was significantly lower than the other two methods. [Conclusion] PBMCs isolated from whole blood by manual separation and NK cells enriched by negative selection with immunomagnetic beads have the potential to provide NK cell materials for CAR-NK cell therapy, and NK cells enriched by platelet lysate-conditioned medium have the potential to provide NK cells for large-scale NK cell activation reinfusion therapy.

Blood resource shortages, characterized by regional imbalance, seasonal fluctuations, and expiration-related wastage, pose a critical challenge to China's public health system. Cryopreserved red blood cell (RBC) technology, with its ultra-long preservation period, transfusion efficacy, and safety, offers a strategic pathway to alleviate chronic blood shortages and optimize resource allocation. This article systematically reviews the technological evolution of RBC cryopreservation, clarifies its core advantages, and identifies bottlenecks and improvement strategies in large-scale application. The establishment of a regularized cryopreserved RBC bank holds multidimensional strategic value; however, its practical implementation requires resolving challenges such as thawing timeliness, standardization gaps, and insufficient grassroots equipment. Future advancements should prioritize glycerol-free washing-free cryopreservation technologies, post-thaw extended preservation additive formulations, and intelligent thawing equipment to overcome throughput and spatiotemporal constraints, thereby providing robust technical support for building a regularized cryopreserved RBC bank and enhancing the efficiency of China's blood security framework.

Objective:To evaluate the therapeutic effect of glycyrrhetinic acid on cough variant asthma (CVA) mice based on molecular docking technique; To explore the possibility of its treatment for cough variant asthma.Methods:The software of Autodock Vina was used for molecular docking. The mice were divided into control group, model group, prednisone acetate group, glycyrrhetinic acid high-, medium-, and low-dosage groups according to the random number table method, with 8 mice in each group. Except for the blank control group, all other groups were induced by egg protein to establish cough variant asthma models. Glycyrrhetinic acid high-, medium-, and low-dosage groups were orally administered glycyrrhetinic acid suspension at 20, 10, and 5 mg/kg, while the prednisone acetate group was orally administered prednisone acetate at 5 mg/kg. The blank control group and model group were orally administered equal volumes of physiological saline, once per day for 14 consecutive days. The animal asthma behavior was observed after drug administration. The secretion of bronchial mucus in lung tissue were observed by AB-PAS staining and the index of spleen were recorded. The protein expressions of Gata3, IL-4 and IL-13 in the spleen tissue were determined by Western blot.Results:Molecular docking results showed that glycyrrhetinic acid had good binding ability to Th2-related factors Gata3, IL-4 and IL-13. Results of animal experiment showed that compared with the model group, the mucus secretion decreased in glycyrrhetinic acid groups, the index of the spleen of mice obviously decreased, protein expression levels of IL-4 and IL-13 in the spleen tissue of mice in glycyrrhetinic acid high-, medium-, and low-dosage groups decreased ( P<0.05), and Gata3 in glycyrrhetinic acid medium- and low-dosage groups decreased ( P<0.05). Conclusion:Glycyrrhetinic acid can correct the shift of Th2 in the immune system of cough variant asthma mice and has a certain therapeutic effect.

Objective:To investigate the protective effect and mechanism of Acronychia pedunculata water extracts on UV-induced light damage of human keratinocytes.Methods:The experiment was conducted from December 2018 to April 2020 in the Guangxi Medical University Laboratory of Genetics. The photoaged keratinocyte model was used, the cells were co-cultured with different concentrations of Acronychia pedunculata water extracts. The cell proliferation rate was detected by CCK-8 method. The levels of reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and total antioxidant capacity (T-AOC) of cells were detected by a test kit. The levels of IL-1β, IL-6 and tumor necrosis factor-alpha (TNF-α) were determined by ELISA.Results:The proliferation of HaCaT cells was promoted by 0.5 mg/L-2.0 mg/L of the extracts. Compared with control group, the proliferation rate of HaCaT cells in the experimental group was significantly increased ( P<0.05). Compared with control group, the contents of ROS was decreased ( F=214.67, P<0.05), MDA was decreased ( F=811.88, P<0.05), SOD was increased ( F=28.95, P<0.05), CAT was increased ( F=213.31, P<0.05), GPX was increased ( F=65.10, P<0.05), T-AOC was increased ( F=305.58, P<0.05), IL-1β was decreased ( F=15.46, P<0.05), IL-6 was decreased ( F=59.2, P<0.05), and TNF-α was decreased ( F=33.13, P<0.05). Conclusions:The extracts of 0.5-2.0 mg/L of Acronychia pedunculata have protective effects on the photoaging cell model, which may be related to the increase of SOD, CAT, GPX and other antioxidant enzymes and the level of T-AOC in photoaging HaCaT cells, and the decrease of ROS, MDA content and the expression of inflammatory cytokines.

Objective:To explore the application of digital acoustic analysis in assessing the risk of aspiration among persons with dysphagia using the Praat speech analysis software.Methods:The swallowing of 46 stroke survivors with dysphagia was studied using video fluoroscopy. Each patient was required to pronounce " yi/i/" 3 times before and after taking each mouthful of food, and their voice samples were collected. The subjects were divided into a non-aspiration group of 16 and an aspiration group of 30 based on their penetration-aspiration scale scores. Fundamental frequency, relative average perturbation (RAP), jitter, shimmer, amplitude perturbation quotient and harmonic-to-noise ratio were compared between the two groups before and after taking food.Results:For the non-aspiration group, there was no significant difference in the acoustic data before and after eating. For the aspiration group there were significant differences in fundamental frequency, relative average perturbation and jitter before and after taking food. The average RAP and jitter of the non-aspiration group were significantly better than the aspiration group′s averages before eating. After eating, however, significant differences were observed only in average jitter.Conclusions:Analyzing perturbation and jitter can help to identify persons at risk of aspiration.

Objective:To explore the effect of the pharyngeal pressure feedback training on pharyngeal constriction in persons with swallowing disorders caused by brainstem lesions.Methods:Twenty patients with disordered swallowing caused by a brainstem lesion were randomly divided into a control group and an intervention group, each of 10. Both groups received routine swallowing training including oral sensorimotor training, neuromuscular stimulation and balloon catheter dilation, while the intervention group was additionally provided with two weeks of pharyngeal pressure feedback training. Before and after the treatment, both groups were evaluated in terms of the peak pressure of superior and inferior pharyngeal constriction, endoscopically and also using the functional oral intake scale.Results:There was no significant difference between the two groups in any measure before the intervention. Afterward, both groups had improved significantly by all of the measurements, but the average peak upper pharyngeal pressure, the average intake scale score and the endoscopy results of the intervention group were all significantly better than the control group′s averages.Conclusions:Pharyngeal pressure feedback training can significantly relieve swallowing disorders caused by brainstem lesions.

OBJECTIVE: To discuss the advantages and technical limitations of various molecular genetic techniques in the diagnosis of two infants featuring all-round developmental retardation. METHODS: The two patients were initially screened by using chromosomal microarray analysis (CMA). For patient 1, his parents were also subjected to CMA analysis, and the data was analyzed by using ChAS and UPD-tool software. For patient 2, methylation-specific PCR (MS-PCR) was carried out. RESULTS: Patient 1 was diagnosed with maternal uniparental disomy (UPD) type Prader-Willi syndrome (PWS) by CMA and UPD-tool family analysis. His chromosomes 15 were of maternal UPD with homology/heterology. Patient 2 was diagnosed with deletion type PWS by combined CMA and MS-PCR. CONCLUSION Correct selection of laboratory methods based on the advantages and limitations of various molecular techniques can help with diagnosis of genomic imprinting disorders and enable better treatment and prognosis through early intervention.

Objective: To explore the sensitivity and specificity of the Chinese version of the volume and viscosity swallowing test with modified safety and effectiveness indicators (VVST-CV) in assessing deglutition disorders. Methods: The solvent, volume, viscosity, and test index of the volume and viscosity swallowing test were modified before it was used to evaluate 89 patients with swallowing disorders. The positive rates of the safety indexes (coughing, voice changes and blood oxygen saturation decline at least 3%) and the efficiency indexes (poor lip closure, oral residue, pharynx residue and swallowing) were recorded and compared and compared with the positive rates of penetration, aspiration and food residue determined by means of according to the videofluoroscopic swallowing study, then the sensitivity, specificity, positive predictive value and negative predictive value of the modified VVST-CV with regard to the safety and efficiency of swallowing were calculated. Results: The VVST-CV′s sensitivity in detecting dysphagia was 0.97. For impaired safety it was 0.85 and for impaired swallowing efficiency it was 0.95. The sensitivity of the coughing index was 0.65, that of sound change was 0.60 and that of oxygen saturation decrease was 0.42. The positive predictive values for coughing, sound change and oxygen saturation decrease were 0.98, 0.94 and 0.94, respectively. Conclusion The VVST-CV is simple, feasible and sensitive. It can be widely useful in the evaluation of dysphagia in clinical practice.

Totally 21 batches of Gastrodiae Rhizoma pieces from various habitats with different appearance characteristics were analyzed. Five active components( gastrodin,4-hydroxybenzyl alcohol,parishin B,parishin C,and parishin A) were determined by UPLC. Polysaccharide content was determined by phenol sulfuric acid method. And the content of alcohol extracts was determined. The correlation between appearance characteristics and active components,polysaccharide content and amount of alcohol extracts of Gastrodiae Rhizoma pieces was statistically analyzed by SPSS 19. 0 software. And the five active components in the 21 Gastrodiae Rhizoma pieces were clustered by using the Ward' s method in SPSS 19. 0 software. The study found that inconspicuous keratinous Gastrodiae Rhizoma had a low content of each component,and the degree of keratin was positively correlated with the content of each component and the extract,with no significant correlation with the polysaccharide content. In the cluster analysis,the 21 batches were divided into three groups,except the second group whose gastrodin content was significantly higher than the special conditions of other groups. According to the content of the five active components,19 batches of Gastrodiae Rhizoma pieces were classified into two groups with obvious keratin and no obvious keratin. In terms of color,with the deepening of brown color,the content of each component showed a downward trend. The grading of decoction pieces of Gastrodiae Rhizoma shall give full consideration to the color and texture. And the combination of chemical composition and appearance characteristics is an indicator to evaluate the product specification standards.
Drugs, Chinese Herbal ; Gastrodia ; Rhizome

An analysis method was established by UPLC fingerprint and then applied to simultaneous determination of multiple compounds in Gardeniae Fructus from different areas in China. Samples were separated on a Waters Acquity UPLC BEH C₁₈( 2. 1 mm×50 mm,1. 7 μm) column with 0. 1% formic acid-water and acetonitrile solution as gradient mobile phase at a flow rate of 0. 4 m L·min-1 at various wavelengths. The similarity of samples was over 0. 95 with ″Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine( 2012 edition) ″. The UPLC common fingerprints for 32 batches were established with 19 common peaks identified. The samples were divided into 3 groups analyzed by HCA and PCA. Five components were identified as the main compositions which caused the differences of chemical constituents in the samples from different areas with partial least squares discriminant analysis( PLS-DA). The content of the total components in each area was Zhejiang > Fujian > Jiangxi > Sichuan. This method was accurate and viable,could be used to evaluate the quality of Gardeniae Fructus.
China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal/standards* ; Fruit/chemistry* ; Gardenia/chemistry* ; Phytochemicals/analysis*