Objective: To establish an accurate and rapid typing method for Rh typing of samples from patients who have received recent blood transfusions by utilizing the difference in osmotic fragility between fresh and old red blood cells. Methods: A lysing solution suitable for destroying old RBCs was prepared. Sixty-one samples collected in our hospital in 2024 with Rh typing of double groups were treated with the lysing solution to remove the old allogeneic red blood cells while preserving the patient's own fresh red blood cells, followed by repeat Rh typing tests. Results: For 61 samples with Rh typing in double groups, 41 were accurately detected identified through the red blood cell lysis method, yielding an identification rate of 67.21%. No significant difference was observed compared to the detection rate of the commonly used capillary centrifugation modified method (χ =0.103, P>0.05). Conclusion: The red blood cell lysis method provides a novel and rapid experimental approach for clinical use in processing Rh-typed samples that are of double groups, thereby offering a basis for Rh compatibility blood transfusion.

ObjectiveTo explore the effect of serum containing Zhenwutang on myocardial mast cell apoptosis induced by angiotensin Ⅱ （AngⅡ） and the mechanism of the correlation between apoptosis and mitochondrial autophagy. MethodsIn this experiment， AngⅡ and serum containing Zhenwutang with different concentrations were used to interfere with H9C2 cardiomyocytes for 24 h， and the survival rate of H9C2 cardiomyocytes was detected by cell counting kit-8 （CCK-8） to screen the optimal concentration for the experiment. Enzyme-linked immunosorbent assay （ELISA） was used to detect the content of B-type natriuretic peptide （BNP） in cell culture supernatant， and immunofluorescence was used to detect the cell surface area to verify the construction of the myocardial mast cell model. Subsequently， the experiment was divided into a blank group （20% blank serum）， a model group （20% blank serum + 5×10^-5 mol·L^-1 AngⅡ）， low-， medium-， and high-dose （5%， 10% and 20%） serum containing Zhenwutang groups， an autophagy inhibitor group （1×10^-4 mol·L^-1 3-MA）， and autophagy inducer group （1×10^-7 mol·L^-1 rapamycin）. The apoptosis level of H9C2 cells and the changes of mitochondrial membrane potential were detected by flow cytometry. The lysosomal probe （Lyso Tracker） and mitochondrial probe （Mito Tracker） co-localization was employed to detect autophagy. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect Caspase-3， Caspase-9， B-cell lymphoma 2 （Bcl-2）， Bcl-2-related X protein （Bax）， and cytochrome C （Cyt C） in apoptosis-related pathways and the relative mRNA expression of ubiquitin ligase （Parkin）， phosphatase and tensin homolog （PTEN）-induced kinase 1 （PINK1）， and p62 protein in mitochondrial autophagy-related pathways. Western blot was used to detect cleaved Caspase-3， cleaved Caspase-9， Bax， Bcl-2， and Cyt C in apoptosis-related pathways， phosphorylated ubiquitin ligase （p-Parkin）， phosphorylated PTEN-induced kinase 1 （p-PINK1）， p62， and Bcl-2 homology domain protein Beclin1 in mitochondrial autophagy-related pathways， and the change of microtubule-associated protein 1 light chain 3 （LC3） Ⅱ/Ⅰ ratio. ResultsCCK-8 showed that when the concentration of AngⅡ was 5×10^-5 mol·L^-1， the cell activity was the lowest， and there was no cytotoxicity. At this concentration， the surface area of cardiomyocytes was significantly increased （P<0.01）， and the content of BNP in the supernatant of culture medium was significantly increased （P<0.05）. Therefore， AngⅡ with a concentration of 5×10^-5 mol·L^-1 was selected for the subsequent modeling of myocardial mast cells. Compared with the blank group， the model group and the autophagy inhibitor 3-MA group had a significantly increased apoptosis rate （P<0.01） and significantly decreased mitochondrial membrane potential （P<0.01）. The results of immunofluorescence co-localization showed that compared with the blank group， the model group had a significantly decreased number of red and green fluorescence spots. The results of Real-time PCR showed that compared with that in the blank group， the relative mRNA expression of Bax， Caspase-3， Caspase-9， Cyt C， and p62 in the model group was significantly up-regulated （P<0.01）， while the relative mRNA expression of Bcl-2， Parkin， and PINK1 was significantly down-regulated （P<0.01）. In addition， the relative protein expression of Bax， cleaved Caspase-3， cleaved Caspase-9， Cyt C， and p62 was significantly up-regulated （P<0.01）. The LC3Ⅱ/Ⅰ was significantly decreased， and the relative protein expression of Bcl-2， p-Parkin， p-PINK1， and Beclin1 was significantly down-regulated （P<0.01）. Compared with the model group， the serum containing Zhenwutang groups and the autophagy inducer group had significantly decreased apoptosis rate （P<0.01）， and the decrease ratio of mitochondrial membrane potential is significantly lowered （P<0.01） in a dose-dependent manner. Additionally， both red and green fluorescence spots became more in these groups. In the 3-MA group， the number of red and green fluorescence spots decreased significantly. The relative mRNA expression of Bax， Caspase-3， Caspase-9， Cyt C， and p62 was significantly down-regulated （P<0.05， P<0.01）， while that of Bcl-2， Parkin， and PINK1 was significantly up-regulated （P<0.01）. In the serum containing Zhenwutang groups， the relative protein expression levels of Bax， cleaved Caspase-3， cleaved Caspase-9， Cyt C， and p62 were significantly down-regulated （P<0.05，P<0.01）. The LC3Ⅱ/Ⅰ was significantly increased， and the relative protein expression levels of Bcl-2， p-Parkin， p-PINK1， and Beclin1 were significantly up-regulated （P<0.01）. ConclusionThe serum containing Zhenwutang can reduce the apoptosis of myocardial mast cells and increase mitochondrial autophagy. This is related to the inhibition of intracellular Bax/Bcl-2/Caspase-3 apoptosis pathway and regulation of Parkin/PINK1 mitochondrial autophagy pathway.

OBJECTIVE To prepare reactive oxygen species （ROS）-responsive methotrexate （MTX）-modified paclitaxel （PTX）/icariin （ICA） micelles （MTX-oxi-Ms@PTX/ICA）， and perform technology optimization and in vitro anti-tumor effect evaluation. METHODS Synergistic toxicity concentration range of PTX and ICA was screened by synergistic toxicity test. The micelles were prepared by thin film hydration method， and their technology was optimized by response surface methodology. The fundamental characteristics of the micelles prepared by the optimal technology were evaluated. The micelles’ cytotoxicity， targeting ability to renal carcinoma RENCA cells of mice， and their inhibitory effects on invasion and migration were assessed. RESULTS Results of synergistic toxicity experiments demonstrated that the strongest synergistic effect occurred when PTX concentrations ranged from 2.5 to 10 μmol/L and ICA concentrations ranged from 5 to 15 μmol/L. The optimal technology of MTX-oxi-Ms@PTX/ ICA was determined to include 80 mg Soluplus®， Soluplus® and TPGS1000 mass ratio of 4∶1 （mg/mg）， 2 mg DSPE-PEG2000-TK- PEG5000， 2 mg DSPE-PEG2000-MTX， 1 mg PTX， and 1.5 mg ICA， with a hydration temperature of 35 ℃ and a formulation volume of 5 mL. Under the optimal conditions， average encapsulation efficiency of PTX and ICA in 3 batches of MTX-oxi- Ms@PTX/ICA reached 92.75%， the critical micelle concentration （CMC） was 0.007 9 mg/mL， the particle size was （62.09±1.68） nm， the polydispersity index （PDI） was 0.046±0.032， and the Zeta potential was （－2.47±0.15） mV. Within 30 days of placement， there was no significant change E-mail：yingqiang_1126@163.com in particle size and polydispersity index of micelle. In vitro release experiments showed that MTX-oxi-Ms@PTX/ICA released drugs more rapidly in oxidative environments. The half maximal inhibitory concentration of MTX-oxi-Ms@PTX/ICA against RENCA cells was （5.170±0.036） μmol/L. In vitro cellular uptake experiments indicated that compared with unmodified micelles， MTX modified micelles had stronger targeting effects on cancer cells， and also significantly enhanced the inhibitory ability of invasion and migration of RENCA cells （P＜0.05）. CONCLUSIONS MTX-oxi-Ms@PTX/ICA micelles are successfully prepared， which exhibit high encapsulation efficiency， low critical micelle concentration， and good stability. These micelles demonstrate significant cytotoxicity against RENCA cells and effectively inhibit cancer cell invasion and migration.

Objective To study the death status of pancreatic cancer among residents in Xuhui district, Shanghai, from 1992 to 2021, and analyze its trends of change, so as to provide evidence for the prevention and treatment of pancreatic cancer. Methods Based on the database of Shanghai death registration system from 1992 to 2021, the crude mortality rate, standardized mortality rate, age-specific mortality rate and other indicators of pancreatic cancer among registered residents in Xuhui district were calculated. The Joinpoint software was used to analyze the trends of average annual percent change (AAPC) of pancreatic cancer mortality rate, and the age-period-cohort model was used to analyze the age effect, period effect and birth cohort effect pairs significant changes in pancreatic cancer mortality. Results In 2021, the mortality rate of pancreatic cancer in Xuhui district, Shanghai, ranked fourth among malignant tumors, and the winning rate and world standard rate of the whole population, males and females were 8.34/100 000 (8.81/100 000, 7.98/100 000) and 7.28/100 000 (7.69/100 000, 6.96/100 000), respectively, with males higher than females. AAPC of crude mortality rate and the standardized (6) mortality rate were higher in males than that in females. The age-specific mortality rate increased with the increase of age, and the highest mortality rate was found in 60-84 years old group. The age-period-cohort model showed that from 1992 to 2021, the annual net shift of pancreatic cancer mortality among the whole population, male and female residents in Xuhui district, Shanghai, was 1.22%, 1.58%, 1.15% (P=0.20, 0.19, 0.45) respectively, and the time trend was not significant. From the perspective of age effect, the risk of death from pancreatic cancer in the whole population and with age deviation in males had an obvious trend with increasing age (P<0.05), while the age effect in females had no obvious trend. From the perspective of period effect, no period deviation was significant in the whole population, males and females (P>0.05). In terms of cohort effects, there were significant differences in the whole population and the male cohort deviations(P<0.05). No significant cohort effect was observed in the female population. Conclusions The mortality rate of pancreatic cancer among registered residents in Xuhui district, Shanghai from 1992 to 2021, was on the rise, especially in the 60-84 years old group and male. The prevention and control of pancreatic cancer needs to develop effective epidemic prevention measures for corresponding populations.

Objective:To evaluate the clinical efficacy of Buyang Huanwu Decoction combined with medium-frequency pulsed electrical stimulation in the treatment of diabetic peripheral neuropathic pain (DPNP).Methods:A randomized controlled trial study was conducted. A total of 86 patients with DPNP in the endocrinology department of Xi'an Hospital of Traditional Chinese Medicine from September 2020 to September 2023 were selected as the observation objects and divided into 2 groups through the random number table method, with 43 cases in each group. The control group was combined with medium-frequency pulsed electrical stimulation on the basis of conventional Western medical treatment, while the observation group was supplemented with Buyang Huanwu Decoction on the basis of the control group. Both groups were treated for 12 weeks. TCM syndrome scores were conducted before and after the treatment respectively. The Toronto Clinical Scoring System (TCSS) was used to grade the severity of neuropathy. The motor nerve conduction velocity (MCV) and sensory nerve conduction velocity (SCV) of the median nerve and common peroneal nerve were detected by electromyography evoked potential tester. Adverse reactions during the treatment were recorded to evaluate the clinical efficacy.Results:The total effective rate was 93.02% (40/43) in the observation group and 74.42% (32/43) in the control group, with statistical significance between the two groups ( χ2=5.46, P=0.019). After treatment, the scores of shortness of breath and fatigue, end-limb pain, listlessness, dull tongue and the total score in the observation group were lower than those in the control group ( t values were 3.06, 3.42, 3.09, 3.57 and 3.98 respectively, P<0.05); the scores of neurological symptoms, neural reflexes and sensory function examinations were lower than those in the control group ( t values were 4.54, 6.28 and 5.16 respectively, P<0.01). After treatment in the observation group, MCV of the median nerve [(47.96±5.98) m/s vs. (42.02±5.74) m/s, t=4.70] and SCV [(54.90±6.29) m/s vs. (48.04±6.16) m/s, t=5.11] were higher than those in the control group. The MCV of the common perbular nerve [(41.76±6.27) m/s vs. (37.04±6.18) m/s, t=3.52] and SCV [(50.35±6.41) m/s vs. (46.14±6.13) m/s, t=3.12] were higher than those in the control group ( P<0.01 or P<0.05). During the treatment period, the incidence of adverse reactions was 11.63% (5/43) in the observation group and 9.30% (4/43) in the control group, without statistical significance between the two groups ( χ2=0.12, P=0.725). Conclusion:Buyang Huanwu Decoction combined with medium-frequency pulsed electrical stimulation can effectively improve the degree of neuropathy in patients with DPNP, increase nerve conduction velocity, alleviate TCM syndromes, enhance clinical efficacy and has good safety.

Objective     To investigate the accuracy of 18F-FDG positron emission tomography/computed tomography (PET/CT) combined with CT three-dimensional reconstruction (CT-3D) in the differential diagnosis of benign and malignant pulmonary nodules. Methods     The clinical data of patients who underwent pulmonary nodule surgery in the Department of Thoracic Surgery, Northern Jiangsu People's Hospital from July 2020 to August 2021 were retrospectively analyzed. The preoperative 18F-FDG PET/CT and chest enhanced CT-3D and other imaging data were extracted. The parameters with diagnostic significance were screened by the area under the receiver operating characteristic (ROC) curve (AUC). Three prediction models, including PET/CT prediction model (MOD PET), CT-3D prediction model (MOD CT-3D), and PET/CT combined CT-3D prediction model (MOD combination), were established through binary logistic regression, and the diagnostic performance of the models were validated by ROC curve. Results     A total of 125 patients were enrolled, including 57 males and 68 females, with an average age of 61.16±8.57 years. There were 46 patients with benign nodules, and 79 patients with malignant nodules. A total of 2 PET/CT parameters and 5 CT-3D parameters were extracted. Two PET/CT parameters, SUVmax≥1.5 (AUC=0.688) and abnormal uptake of hilar/mediastinal lymph node metabolism (AUC=0.671), were included in the regression model. Among the CT-3D parameters, CT value histogram peaks (AUC=0.694) and CT-3D morphology (AUC=0.652) were included in the regression model. Finally, the AUC of the MOD PET was verified to be 0.738 [95%CI (0.651, 0.824)], the sensitivity was 74.7%, and the specificity was 60.9%; the AUC of the MOD CT-3D was 0.762 [95%CI (0.677, 0.848)], the sensitivity was 51.9%, and the specificity was 87.0%; the AUC of the MOD combination was 0.857 [95%CI (0.789, 0.925)], the sensitivity was 77.2%, the specificity was 82.6%, and the differences were statistically significant (P<0.001). Conclusion     18F-FDG PET/CT combined with CT-3D can improve the diagnostic performance of pulmonary nodules, and its specificity and sensitivity are better than those of single imaging diagnosis method. The combined prediction model is of great significance for the selection of surgical timing and surgical methods for pulmonary nodules, and provides a theoretical basis for the application of artificial intelligence in the pulmonary nodule diagnosis.

Objective To investigate the epidemiological characteristics and mutation spectrum of monogenic diseases in Chinese population through a large-scale,multicenter carrier screening.Methods This study was conducted among a total of 33 104 participants(16 610 females)from 12 clinical centers across China.Carrier status for 223 genes was analyzed using high-throughput sequencing and different PCR methods.Results The overall combined carrier frequency was 55.58%for 197 autosomal genes and 1.84%for 26 X-linked genes in these participants.Among the 16 669 families,874 at-risk couples(5.24%)were identified.Specifically,584 couples(3.50%)were at risk for autosomal genes,306(1.84%)for X-linked genes,and 16 for both autosomal and X-linked genes.The most frequently detected autosomal at-risk genes included GJB2(autosomal recessive deafness type 1A,393 couples),HBA1/HBA2(α-thalassemia,36 couples),PAH(phenylketonuria,14 couples),and SMN1(spinal muscular atrophy,14 couples).The most frequently detected X-linked at-risk genes were G6PD(G6PD deficiency,236 couples),DMD(Duchenne muscular dystrophy,23 couples),and FMR1(fragile X syndrome,17 couples).After excluding GJB2 c.109G>A,the detection rate of at-risk couples was 3.91%(651/16 669),which was lowered to 1.72%(287/16 669)after further excluding G6PD.The theoretical incidence rate of severe monogenic birth defects was approximately 4.35‰(72.5/16 669).Screening for a battery of the top 22 most frequent genes in the at-risk couples could detect over 95%of at-risk couples,while screening for the top 54 genes further increased the detection rate to over 99%.Conclusion This study reveals the carrier frequencies of 223 monogenic genetic disorders in the Chinese population and provides evidence for carrier screening strategy development and panel design tailored to the Chinese population.In carrier testing,genetic counseling for specific genes or gene variants can be challenging,and the couples need to be informed of these difficulties before testing and provided with options for not screening these genes or gene variants.

Objective To investigate the effect and mechanism of long chain non-coding RNA(ln-cRNA)SNHG16 regulating PAR1 on the proliferation,migration and invasion of lung cancer.Methods From March 2020 to August 2021,lung cancer tissues and adjacent tissues of 35 patients with lung cancer were col-lected,and lung cancer cell lines(HCC827,A549,SK-LU-1,A427)and normal lung cell lines(MRC5)were simultaneously cultured.The overexpression model of PAR1(pcDNA-PAR1)was constructed by using the expression vector pcDNA3.1.A549 cells were divided into four groups after transfection:si-SNHG16,si-PAR1,si-SNHG16+pcDNA-PAR1 and control(transfection with si-NC).The expression levels of lncRNA SNHG16 and PAR1 in lung cancer cell lines(HCC827,A549,SK-LU-1,A427),lung cancer tissues and adja-cent tissues were detected by real-time fluorescence quantitative reverse transcription-polymerase chain reac-tion(qRT-PCR),and the transfection efficiency of each group was verified.MTT assay and clonal formation were used to determine the proliferation of cells in each group,flow cytometry was used to detect the apopto-sis of cells in each group,cell scratch and Transwell test were used to detect the migration and invasion ability of cells in each group,and Western blot was used to detect the protein expression of PAR1.Results The ex-pression level of lncRNA SNHG16 in lung cancer tissue was higher than that in adjacent tissues(P＜0.05).The expression level of lncRNA SNHG16 in lung cancer cell lines(HCC827,A549,SK-LU-1,A427)was higher than that in normal lung cell lines(MRC5),with statistical significance(P＜0.05).The results of qRT-PCR showed that the expression level of lncRNA SNHG16 gene was(21.02±0.04)％of the control af-ter transfection of si-SNHG16,the expression level of PAR1 gene was(19.06±0.02)％of the control after transfection of si-PAR1,and the expression level of PAR1 gene was 2.70±0.00 folds of the control after transfection of pcDNA-PAR 1,the difference was statistically significant(P＜0.05).The results of Western blot showed that the expression level of transfected in each PAR1 protein was different from that of the con-trol(P＜0.05).Compared with the control,the activity of cells transfected with si-SNHG16 was lower,the a-bility of clone formation,cell migration and invasion was obviously inhibited,and the apoptosis rate was higher(P＜0.05),while pcDNA-PAR1 could weaken the influence of transfected si-SNHG16 on cell proliferation,apoptosis,migration and invasion(P＜0.05).lncRNA SNHG16 was positively correlated with the expression level of PAR1(r=0.61).Conclusion lncRNA SNHG16 can regulate the occurrence and development of lung cancer by targeting PAR1.

Objective To investigate the predictive value of serum soluble CD40 ligand(sCD40L)and an-giotensin Ⅱ receptor-like 1 endogenous ligand 13(Apelin-13)levels for the short-term prognosis in patients after traumatic brain injury surgery.Methods A total of 89 patients with traumatic brain injury who under-went treatment and surgery in Zhongwu Hospital of Suqian from June 2020 to December 2022 were collected as the research group.A total of 89 healthy individuals who came to Zhongwu Hospital of Suqian during the same period for physical examination were selected as the control group.The clinical data of the study subjects were collected and the expression levels of sCD40L and Apelin-13 in the serum were detected.Pearson method was applied to analyze the correlation between sCD40L and Apelin-13 in the serum of patients in the research group.Receiver operating characteristic(ROC)curve was applied to analyze the predictive value of serum sCD40L and Apelin-13 for the short-term prognosis in patients after traumatic brain injury surgery.Results Compared with the control group,the serum level of sCD40L in the research group was significantly increased,and the serum level of Apelin-13 was significantly decreased(P<0.05).There were no statistically significant differences in age and gender between the poor prognosis group and the good prognosis group(P>0.05).The proportions of patients with preoperative hospitalization time≥10 h,surgical duration≥4 h,intraoperative bleeding vol-ume≥400 mL,and permanent implants were higher than those in the good prognosis group,and the Glasgow Outcome Scale(GOS)score was lower than that in the good prognosis group(P<0.05).The serum sCD40L level in good prognosis group was lower than that in poor prognosis group,and the serum Apelin-13 level was higher than that in poor prognosis group(P<0.05).The area under the curve(AUC)of serum sCD40L and Apelin-13 levels for predicting the short-term prognosis in patients after traumatic brain injury surgery was 0.776 and 0.819,respectively,with sensitivity of 79.31%and 75.86%,specificity of 75.00%and 81.67%,respectively,and the AUC of the combination of the two was 0.909,with sensitivity of 89.66%and specificity of 75.00%,respectively.Conclusion The serum sCD40L level increases and the serum Apelin-13 level decrea-ses in patients with poor short-term prognosis after traumatic brain injury surgery.The combined detection of the two has high predictive value for the short-term prognosis in patients.

Objective To investigate the epidemiological characteristics and mutation spectrum of monogenic diseases in Chinese population through a large-scale,multicenter carrier screening.Methods This study was conducted among a total of 33 104 participants(16 610 females)from 12 clinical centers across China.Carrier status for 223 genes was analyzed using high-throughput sequencing and different PCR methods.Results The overall combined carrier frequency was 55.58%for 197 autosomal genes and 1.84%for 26 X-linked genes in these participants.Among the 16 669 families,874 at-risk couples(5.24%)were identified.Specifically,584 couples(3.50%)were at risk for autosomal genes,306(1.84%)for X-linked genes,and 16 for both autosomal and X-linked genes.The most frequently detected autosomal at-risk genes included GJB2(autosomal recessive deafness type 1A,393 couples),HBA1/HBA2(α-thalassemia,36 couples),PAH(phenylketonuria,14 couples),and SMN1(spinal muscular atrophy,14 couples).The most frequently detected X-linked at-risk genes were G6PD(G6PD deficiency,236 couples),DMD(Duchenne muscular dystrophy,23 couples),and FMR1(fragile X syndrome,17 couples).After excluding GJB2 c.109G>A,the detection rate of at-risk couples was 3.91%(651/16 669),which was lowered to 1.72%(287/16 669)after further excluding G6PD.The theoretical incidence rate of severe monogenic birth defects was approximately 4.35‰(72.5/16 669).Screening for a battery of the top 22 most frequent genes in the at-risk couples could detect over 95%of at-risk couples,while screening for the top 54 genes further increased the detection rate to over 99%.Conclusion This study reveals the carrier frequencies of 223 monogenic genetic disorders in the Chinese population and provides evidence for carrier screening strategy development and panel design tailored to the Chinese population.In carrier testing,genetic counseling for specific genes or gene variants can be challenging,and the couples need to be informed of these difficulties before testing and provided with options for not screening these genes or gene variants.