[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.

BACKGROUND:Apoptosis plays an important role in secondary brain injury.Therefore,to explore the pathophysiological mechanism of promoting nerve cell survival after traumatic brain injury provides a new direction and theoretical basis for the prevention and treatment of traumatic brain injury. OBJECTIVE:To explore the expression changes of Lnx1 molecule in mammalian cortical neurons after brain injury and the possible mechanism involved in secondary brain injury. METHODS:Eighty adult SD rats were divided into 20 male and 20 female mice in sham operation group and 20 male and 20 female mice in traumatic brain injury group.The traumatic brain injury rat model was established by heavy falling method.At 6,12,24,48,and 72 hours after brain injury,the expression of related molecules in damaged cortical neurons was analyzed by RT-qPCR,western blot assay,and immunofluorescence staining. RESULTS AND CONCLUSION:(1)The brain tissue of traumatic brain injury group was bleeding and obvious tissue injury could be observed.Water content of brain tissue increased after traumatic brain injury.(2)Compared with the sham operation group,the expression of Lnx1 in cortical neurons after traumatic brain injury increased significantly at 24 hours after injury.(3)After traumatic brain injury,the expression of PBK and BCR protein decreased,and the pro-survival factor ctgf increased.(4)These findings suggest that after traumatic brain injury,the expression of Lnx1 is up-regulated in neurons,which may be due to the decrease of the expression of its target molecules PBK and BCR,and further promote the expression of living factor ctgf,which has a protective effect on the damaged neurons.

Objective To establish a culture method for micropapillary lung adenocarcinoma organoids and conduct targeted drug screening.Methods Organoids were extracted and cultured from a surgical tissue sample of a patient diagnosed with micropapillary lung adenocarcinoma,and the growth of lung cancer organoids was observed and recorded dynamically.The morphological and gene expression characteristics of tumor cells between lung cancer organoids and parental tissue were compared using hematoxylin eosin(HE)staining and immunohistochemical methods.Real time fluorescence quantitative polynucleotide chain reaction(qRT-PCR)method was used to detect gene mutations in lung cancer parental tissue and organoids.Finally,based on results of genetic testing,targeted drugs were selected and their therapeutic effects were verified.Results We have successfully cultured spherical organoids from micropapillary lung adenocarcinoma tissue,which can be passaged for at least 3 generations.HE staining results showed that the morphology of tumor cells in organoids was roughly consistent with that of parental tissue.The immunohistochemical results showed that the protein expression levels of various genes in lung cancer organoids and parental tissue were roughly the same.Results of gene mutation analysis showed that the mutated genes in lung cancer parental tissue and organoids were consistent,both reflecting RET fusion.The screening results of targeted drugs based on lung cancer organoids showed that vandertinib had the best anti-tumor effect in vitro.Conclusion Drug screening experiments based on micropapillary lung adenocarcinoma organoids can screen highly efficient targeted drugs in a short period of time,which may benefit patients with micropapillary lung adenocarcinoma.

Objective:To summarize the clinical features of primary segmental volvulus (PSV) in neonates.Methods:A retrospective analysis was conducted on the clinical data of neonates with PSV who were admitted to the Department of Neonatal Surgery, Children's Hospital Affiliated to Capital Institute of Pediatrics from May 2014 to May 2023. The clinical manifestations, auxiliary examinations, treatment and prognosis of the neonates were summarized, and descriptive statistical analysis was performed on the collected data.Results:A total of 10 neonates with PSV were included, with a mean gestational age of (34.1±3.0) weeks and birth weight of (2 291±646) g. Eight cases had an onset age of 3 d or less, and 2 cases had an onset age of more than 3 d. Abdominal distension was observed as the main manifestation in all cases, while bilious vomiting occurred in seven cases and hematochezia in five cases. Imaging examinations mainly revealed low intestinal obstruction without specific manifestations. Laboratory tests showed metabolic acidosis and varing degrees of anaemia. Nine cases underwent diagnostic abdominal puncture, of which five had bloody ascites, two had clear ascites, one had bloody mixed with fecal-like ascites, and one had chylous ascites. All the cases underwent emergency exploratory laparotomy and segmental small bowel resections with either primary intestinal anastomosis or enterostomy. All cases were successfully cured and had been followed up to the age of 4 months to 9 years with good growth and development as normal children of the same age.Conclusions:Neonatal PSV is an independent abdominal emergency characterized by non-specific clinical manifestations and difficult preoperative diagnosis, but the overall prognosis is favorable after active surgical treatment.

Objective Based on sentinel symptoms,a nursing intervention program for gastrointestinal symptom group of postoperative chemotherapy for lung cancer was constructed and its application effect was evaluated.Methods The nursing intervention program of gastrointestinal symptom group was constructed on the basis of ref-erence guidelines,qualitative interview and expert consultation.From January 2021 to January 2022,a total of 330 patients with postoperative chemotherapy for lung cancer in a tertiary hospital in Shenyang were selected as re-search subjects.The experimental group received the gastrointestinal symptoms group nursing intervention program on the basis of routine nursing,and the control group received routine care.Patients were investigated with the M.D.Anderson Symptom Inventory and the MOS 36-Item Short-Form Health Survey before 1st chemotherapy(T1),3rd chemotherapy(T2)and 5th chemotherapy(T3).Results After the intervention,the total scores of the 2 groups and the total scores of each symptom in T2 and T3 were statistically significant(P＜0.05),and the score of the experi-mental group was lower than that of the control group.For the scores of 6 dimensions of physiological function,physical pain,overall health,vitality,emotional function,mental health in 2 groups between different time points,the differences are statistically significant(P＜0.05).Conclusion The nursing intervention program of the gastroin-testinal symptom group based on sentinel symptoms is beneficial to reduce the severity of the gastrointestinal symp-tom group and improve the quality of life for postoperative chemotherapy for lung cancer patients.

Objective To investigate the effect of adenotonsillectomy on immune function and sleep structure in children with obstructive sleep apnea-hypopnea syndrome(OSAHS).Methods A total of 94 children with OSAHS treated at Children's Hospital Affiliated to Zhengzhou University from November 2020 to June 2022 were selected as the observation group,and another 80 healthy children who underwent physical examinations at the same hospital during the same period were selected as the control group.Children in the observation group underwent bilateral tonsillectomy combined with endoscopic adenoidectomy.Changes in the sleep structure of children in the observation group were detected before surgery and at discharge by using a multi-channel sleep monitoring system.Venous blood samples were collected from children in the observation group before surgery,one month after surgery,and six months after surgery,while venous blood samples of children in the control group were collected on the day of physical examination.The serum levels of IgA,IgG and IgM were measured by using immunoturbidimetry,the percentages of CD3+,CD4+and CD8+in the plasma were measured by using flow cytometry,and the CD4+/CD8+ratio was calculated.Results Compared with before surgery,the proportion of non-rapid eye movement sleep(NREM)1,apnea-hypopnea index and obstructive apnea-hypopnea index of children in the observation group decreased at discharge,the proportions of NREM2,NREM3 and rapid eye movement sleep(REM),sleep efficiency and the lowest oxygen saturation increased,and REM time extended(P＜0.05).The preoperative serum levels of IgA,IgG and IgM in the observation group were significantly higher than those in the control group(P＜0.05).One month after surgery,the serum levels of IgA,IgG and IgM in the observation group were significantly lower than those in the control group(P＜0.05).There was no statistically significant difference in the serum levels of IgA,IgG and IgM between the observation group and the control group at six months after surgery(P＞0.05).One and six months after surgery,the serum levels of IgA,IgG and IgM in the observation group were significantly lower than those before surgery(P＜0.05),and the serum levels of IgA,IgG and IgM at six months after surgery were significantly higher than those one month after surgery(P＜0.05).The preoperative CD3+and CD4+percentages and the CD4+/CD8+ratio in the observation group were significantly lower than those in the control group,while the CD8+percentage was significantly higher than that in the control group(P＜0.05).One month after surgery,the CD3+and CD4+percentages and the CD4+/CD8+ratio in the observation group were significantly lower than those in the control group,while the CD8+percentage was significantly higher than that in the control group(P＜0.05).There was no statistically significant difference in the CD3+,CD4+and CD8+percentages and the CD4+/CD8+ratio between the observation group and the control group at six months after surgery(P＞0.05).One month after surgery,the CD3+and CD4+percentages and the CD4+/CD8+ratio in the observation group were significantly lower than those before surgery,while the CD8+percentage was significantly higher than that before surgery(P＜0.05);six months after surgery,the CD3+and CD4+percentages and the CD4+/CD8+ratio in the observation group were significantly higher than those before surgery,while the CD8+percentage was significantly lower than that before surgery(P＜0.05).The CD3+and CD4+percentages and the CD4+/CD8+ratio at six months after surgery were significantly higher than those at one month after surgery,while the CD8+percentage was significantly lower than that at one month after surgery in the observation group(P＜0.05).Conclusion Adenotonsillectomy can effectively prolong the REM time of OSAHS children and improve their sleep efficiency,sleep structure and the immune regulation function of the body.The immune function of the body decreases briefly after adenotonsillectomy,and then gradually returns to the normal level.

Objective To investigate the risk factors for pulmonary embolism (PE)in patients with acute spinal cord injury (ASCI).Methods A case-control design was adopted in this study.Basic information,injury cause,injury site,imaging and laboratory examination results and other relevant data of 840 ASCI patients admitted to 3 affiliated hospitals of Army Medical University from January 2018 to August 2023 were collected.According to occurrence of PE or not,these patients were divided a PE group (n=83) and a non-PE group (n=757).Univariate analysis was performed on 37 risk factors including gender,age,length of hospital stay,length of intensive care unit (ICU)stay,cause of injury,and so on,and then multivariate logistic regression analysis was applied to analyze these selected significant variables.Results Univariate analysis showed that 24 factors such as age,length of hospital stay,lower limb fracture,pulmonary contusion and infection,shock and number of operations were related to the occurrence of PE in ASCI patients (P<0.05).Binary logistic regression analysis indicated that age (P=0.000,OR=1.957,95％CI:1.532～2.499),length of ICU stay (P=0.000,OR=2.561,95％CI:1.728～3.794),number of operations (P=0.015,OR=1.814,95％CI:1.124～2.927),spinal fixation (P=0.004,OR=0.370,95％CI:0.189～0.724),restrictive position (P=0.000,OR=21.269,95％CI:5.276～85.740),platelet count (P=0.000,OR=1.553,95％CI:1.236～1.951),D-dimer level (P=0.000,OR=1.600,95％CI:1.376～1.860)and prothrombin time (P=0.025,OR=2.756,95％CI:1.138～6.670)were independent risk factors for PE in the patients.Conclusion The mortality of the ASCI patients with PE is significantly higher than those without.Age,length of ICU stay,number of operations,spinal fixation,restrictive position,platelet count,D-dimer level and prothrombin time are closely associated with PE in ASCI patients.

Objective:To investigate the clinicopathological features and differential diagnosis of eosinophilic vacuolated tumor (EVT).Methods:Seven cases of EVT with characteristic morphology and unequivocal diagnosis from the Affiliated Hospital of Qingdao University (6 cases), Qingdao, China and the 971 Hospital of PLA Navy (1 case), Qingdao, China between January 2010 and December 2021 were subject to morphological and immunohistochemical analyses. Additionally, whole exome sequencing (WES) was performed in two cases. Twenty-two cases of renal oncocytoma (RO) and 17 cases of eosinophilic chromophobe renal cell carcinoma (eChRCC) diagnosed at the same time were used as controls.Results:Four males and three females with a mean age of 42 years (range: 29-61 years) were included in the study. The tumors were nodular and well-circumscribed, with sizes ranging from 1.5 to 4.5 cm. On cross-section, they appeared gray-red or gray-white, solid, and soft. Tumor cells were arranged in nests, solid sheets, and acinar or small vesicular structures. These cells exhibited eosinophilic cytoplasm with large, prominent clear vacuoles and round nuclei with prominent nucleoli. Perinuclear halos were focally present in four cases, while small tumor cells with sparse cytoplasm and hyperchromatic nuclei were seen in one case. No necrosis or mitosis was noted. Edematous stroma was detected in three cases. All tumors were positive for CD117 and Cathepsin K, but negative for vimentin and CK7. CK20 was positive in scattered individual cells, and Ki-67 positivity ranged from 1% to 4%. Point mutations in MTOR were identified in both patients who were subject to the molecular analysis. Statistical differences in the expression of Cathepsin K, CD10, S-100A1, and Cyclin D1 between EVT and RO ( P<0.05) were significant, so were the differences in the expression of Cathepsin K, CD10, CK7 and claudin 7 between EVT and eChRCC ( P<0.001). Seven patients were followed up for 4 to 96 months (mean, 50 months), with no recurrences or metastases. Conclusions:EVT is a rare renal tumor that shares morphological and immunophenotypic features with RO and eChRCC, and it is closely linked to the TSC/MTOR pathway. The presence of large prominent transparent vacuoles in eosinophilic cytoplasm along with conspicuous nucleoli is its key morphological characteristics. The use of combined immunohistochemical stains greatly aids in its diagnosis. Typically, the tumor exhibits indolent biological behaviors with a favorable prognosis.

China has classified the Corona Virus Disease 2019(COVID-19) as a statutory category B infectious disease and managed it according to Category B since January 8, 2023.In view that Omicron variant is currently the main epidemic strain in China, in order to guide the treatment of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) infection in children with the times, refer to the Diagnosis and Treatment Protocol for Novel Coronavirus Infection (Trial 10 th Edition), Expert Consensus on Diagnosis, Treatment and Prevention of Novel Coronavirus Infection in Children (Fourth Edition) and the Diagnosis and Treatment Strategy for Pediatric Related Viral Infections.The Expert Consensus on the Diagnosis, Treatment and Prevention of Novel Coronavirus Infection in Children (Fifth Edition) has been formulated and updated accordingly on related etiology, epidemiology, pathogenic mechanism, clinical manifestations, auxiliary examination, diagnosis and treatment, and added key points for the treatment of COVID-19 related encephalopathy, fulminating myocarditis and other serious complications for clinical reference.

Nasopharyngeal carcinoma (NPC) is the third most common malignancy with a high recurrence and metastasis rate in South China. Natural compounds extracted from traditional Chinese herbal medicines have been developed and utilized for the treatment of a variety of cancers with modest properties and slight side effects. Maackiain (MA) is a type of flavonoid that was first isolated from leguminous plants, and it has been reported to relieve various nervous system disorders and exert anti-allergic as well as anti-inflammatory effects. In this study, we demonstrated that MA inhibited proliferation, arrested cell cycle and induced apoptosis in nasopharyngeal carcinoma CNE1 and CNE2 cells in vitro and in vivo. The expression of the related proteins associated with these processes were consistent with the above effects. Moreover, transcriptome sequencing and subsequent Western blot experiments revealed that inhibition of the MAPK/Ras pathway may be responsible to the anti-tumor effect of MA on NPC cells. Therefore, the effects of MA and an activator of this pathway, tertiary butylhydroquinone (TBHQ), alone or combination, were investigated. The results showed TBHQ neutralized the inhibitory effects of MA. These data suggest that MA exerts its anti-tumor effect by inhibiting the MAPK/Ras signaling pathway and it has the potential to become a treatment for patients with NPC.
Humans ; Nasopharyngeal Carcinoma/pathology* ; Cell Line, Tumor ; Cell Proliferation ; Apoptosis ; Signal Transduction ; Nasopharyngeal Neoplasms/pathology*