Objective: To create a prediction model for stroke risk among middle-aged and elderly populations, so as to provide a basis for early identification of high-risk population for stroke. Methods: From October to December 2023, residents aged ≥45 years in Chongchuan District, Nantong City, Jiangsu Province were selected using a multi-stage stratified random sampling method. The demographic information, life behavior, and chronic disease data were collected through a questionnaire survey. The standardized prevalence of stroke was calculated using data from the seventh National Population Census. The subjects were randomly divided into the training set and the internal validation set according to the ratio of 8∶2. The basic demographic information, life behavior, and chronic diseases of residents aged ≥45 years in Rugao City were collected from July to August 2023 as the external validation set. Predictive factors were selected using multivariable logistic regression model, and a nomogram for stroke among residents aged ≥45 years was established. The prediction effect was evaluated using the area under the curve (AUC) of the receiver operating characteristic (ROC), calibration curve, and Hosmer-Lemeshow goodness of fit test. Results: A total of 6 290 residents aged ≥45 years were included, including 2 975 males (47.30%) and 3 315 females (52.70%). The average age was (61.90±10.20) years. The prevalence of stroke was 3.80%, and the standardized prevalence was 3.36%. The multivariable logistic regression showed that age, smoking, hypertension, and hyperlipidemia were predictors of stroke risk among residents aged ≥45 years, and the prediction model was ln[p/(1-p)]=-4.619+0.046×age+0.383×smoking+0.887×hypertension+0.678×hyperlipidemia. The AUC values of the training set, internal validation set, and external validation set were 0.748, 0.755, and 0.738, respectively. The consistency indexes were 0.748, 0.755, and 0.738, respectively. The Hosmer-Lemeshow goodness of fit test showed a good fitting effect (P>0.05). Conclusion The prediction model based on age, smoking, hypertension, and hyperlipidemia has good discrimination and calibration, and can be used to predict the risk of stroke among middle-aged and elderly populations aged ≥45 years.

OBJECTIVE To prepare nanoparticle-based targeting preparation loaded with triptolide （TP）， and evaluate its quality， targeting ability and cytotoxic effects. METHODS Polymer nanoparticles carrying TP-targeted folic acid （FA） receptor （TP@PLGA-PEG-FA） were fabricated using poly （lactic-co-glycolic acid）/polyethylene glycol/FA （PLGA-PEG-FA） as the carrier by emulsion and volatilization technique. The morphology and distribution were observed， and their particle size， Zeta potential， polydispersity index （PDI）， drug loading capacity and encapsulation efficiency were measured. Their stability， blood compatibility， in vitro drug release， uptake by RAW264.7 cells （localization with fluorescent dye Cy3.5）， and in vitro cytotoxicity were evaluated. RESULTS TP@PLGA-PEG-FA exhibited spherical shape and uniform distribution， with particle size of （122.60±0.02） nm， Zeta potential of （－17.6±0.6）mV， and PDI of 0.26±0.02； drug loading capacity and encapsulation efficiency of TP were measured to be （7.78±0.05）% and （68.62±0.03）%， respectively. The hemolysis rates of 100， 200， 300， 400 µg/mL TP@PLGA- PEG-FA were 0.77%， 0.92%， 1.34% and 1.63%， respectively. There were no significant changes in particle size， PDI and Zeta potential when TP@PLGA-PEG-FA were placed in 4 ℃ water for 14 days and in DMEM culture medium containing 10% fetal bovine serum at 37 ℃ for 12 h. The cumulative release rate of TP@PLGA-PEG-FA was （84.83±0.29）% in phosphate buffer at pH5.5 for 72 h， which was significantly higher than the cumulative release rates in phosphate buffer solutions at pH7.4 and 6.5 for 72 h （[ 42.37±0.35）% and （63.83±0.29）% ， respectively] （P＜0.05）. Activated RAW264.7 cells took up significantly more Cy3.5@PLGA-PEG-FA than they took up Cy3.5@PLGA-PEG-FA+free FA and Cy3.5@PLGA-PEG. When the mass concentration of TP was≥15.63 ng/mL， the survival rates of activated cells in the TP@PLGA-PEG-FA groups were significantly lower than those of the same mass concentration of free TP groups （P＜0.05）. CONCLUSIONS The prepared TP@PLGA-PEG-FA has high stability， good blood compatibility， active targeting and cytotoxicity to inflammatory cells.

Objective:To investigate the effect of betaine in oxygen-glucose deprivation injury of rat brain microvascular endothelial cells(BMECs),and to clarify the regulatory effect of betaine on phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)pathway.Methods:Five SD rats aged 7 d were selected and the rat BMSEs were obtained.The oxygen-glucose deprivation model of rat BMECs was prepared under hypoxic and hypoglycemic conditions;the experiment was divdided into model group,and low dose,medium dose,and high dose of betaine groups and positive control group,at the same time,blank control group(without modeling)was set up.The BMECs in blank control group and model group were treated with fresh medium,the BMECs in positive control group were given a final concentration of 10 μmnol·L-1 nimodipine,and the BMECs in low,medium and high doses of betaine groups were treated with betaine at final concentration of 0.5,1.0 and 2.0 mmol·L-1,respectively.The survival rates of BMECs in various groups were determined by CCK-8 method at 12,24 and 48 h after culture;the activities of lactate dehydrogenase(LDH)and the levels of adenine ribonucleoside triphosphate(ATP)in the rat BMECs in various groups were determined using kits,and the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL-1β,and IL-18 in supernatants of the BMECs in various groups were determined by enzyme-linked immunosorbent assay(ELISA);the activities of superoxide dismutase(SOD)and levels of malondialdehyde(MDA)in the BMECs in various groups were determined by kits;the transendothelial resistance(TEER)values of rat BMSCs in various groups were determined by TEER analyzer,and the horseradish peroxidase(HRP)permeabilities of BMECs in various groups were determined by an insertion cell culture apparatus.TUNEL staining was used to determine the apoptotic rates of rat BMECs in vaisous groups,and Western blotting method was used to determine the ratios of phosphory lated PI3K(p-PI3K)/PI3K and phosphorylated AKT(p-AKT)/AKT in the rat BMECs in various groups.Results:Compared with blank control group,the survival rate of BMECs,activity of SOD,and level of ATP,value of TEER,and ratios of p-PI3K/PI3K and p-AKT/AKT of the rat BMECs in model group were significantly decreased(P＜0.05),while the activity of LDH,the levels of TNF-α,IL-6,IL-1β,IL-18,and MDA,the apoptotic rate of the BMECs,and HRP permeability were significantly increased(P＜0.05).Compared with model group,the survival rates of the BMECs,activities of SOD,and levels of ATP,values of TEER,and ratios of p-PI3K/PI3K and p-AKT/AKT of the BMECs in low,medium,and high doses of betaine groups and positive control group were significantly increased(P＜0.05),while the activities of LDH,the levels of TNF-α,IL-6,IL-1β,IL-18,and MDA,the apoptotic rates of BMECs and HRP permeabilities were significantly decreased(P＜0.05).Conclusion:Betaine can significantly repair the oxygen-glucose deprivation/reperfusion injury in the rat BMECs,inhibit the oxidative damage and apoptosis of BMECs,and improve the permeability of the cells;its mechanism may be related to the regulation of the PI3K/AKT pathway.

Objective:To explore the mechanism of modified Shaoyao Decoction with Astragali Radix and Houttuynize Herba in regulating PI3K/Akt/NF-κB pathway against intestinal mucosal barrier injury in ulcerative colitis (UC) rats.Methods:Totally 72 Wistar rats were randomly divided into a blank control group, a model group, a mesalazine group, and modified Shaoyao Decoction high-, medium-, and low-dose groups using a random number table method. After successful modeling, the mesalazine group was given 0.2 g/kg of mesalazine suspension by gavage, while modified Shaoyao Decoction high-, medium-, and low-dose groups were given 44.5, 22.3, and 11.1 g/kg of modified Shaoyao Decoction by gavage, once a day, for a total of 4 weeks. The pathological changes of colon tissue were observed using HE staining; immunofluorescence assay was used to detect the expressions of tight junction protein (ZO-1), cytoskeletal protein (F-actin), and vascular endothelial cadherin (VE cadherin) in colon tissue; Western blot was used to detect the protein expressions of PI3K, Akt, NF-κB, VEGF, and cyclooxygenase (COX-2) in colon tissue; QRT PCR was used to detect the expressions of PI3K and Akt mRNA in colon tissue; ELISA method was used to detect the levels of serum TNF-α and endothelin (ET).Results:Compared with the model group, the DAI scores of the mesalazine group and modified Shaoyao Decoction high-dosage group decreased significantly on the 7th, 14th, 21st and 28th day after modeling ( P<0.05 or P<0.01). In mesalazine group and modified Shaoyao Decoction high-, medium-, and low-dose groups, the pathological injury score of colon tissue decreased ( P<0.01), the immunofluorescence intensity of ZO-1, F-actin and VE-cadherin protein expression in colon tissue increased ( P<0.05 or P<0.01), and the expressions of PI3K, Akt, NF-κB and VEGF protein decreased ( P<0.01). The expression of COX-2 protein and the levels of PI3K mRNA and Akt mRNA decreased in the mesalazine group and modified Shaoyao Decoction high- and medium groups ( P<0.05 or P<0.01). Conclusion:Modified Shaoyao Decoction can antagonize the intestinal barrier injury of UC by inhibiting PI3K/Akt/NF-κB pathway and promote the healing of intestinal mucosal ulcer.

【Objective】 To investigate the frequency of platelet antibodies in voluntary blood donors and patients in Zhongshan, Guangdong Province, and to study the specificity and cross-matching of platelet antibodies. 【Methods】 Platelet antibodies of blood donors and patients were screened by solid-phase immunoadsorption (SPIA), rechecked by flow cytometry (FCM), and antibody specificity was identified by PakPlus enzyme immunoassay, and platelet cross-matching was simulated by SPIA. 【Results】 A total of 1 049 blood donor samples and 598 patient samples were tested, with 6 (0.57%) and 49 (8.19%) samples positive for SPIA,respectively(P<0.05); In SPIA positive samples, the positive concordance rate of FCM in blood donors and patients was 100% vs 95%, and that of enzyme immunoassay was 100% vs 88%. Among the initial screening positive samples of blood donors, 5 were anti-HLA Ⅰ antibodies, accounting for 83%, and 1 was anti CD36 antibody, accounting for 17%, with an incidence rate of 0.10%. Among the 14 samples of enzyme immunoassay positive patients, 2 were anti-GP Ⅱb/Ⅲa, 1 was anti-GP Ⅱa/Ⅱa, 8 were anti HLA Ⅰ, and 3 were mixed antibodies (HLA Ⅰ, GP Ⅱb/Ⅲa, GP Ⅰa/Ⅱa). According to the types of antibodies, HLA Ⅰ antibodies were the most common, accounting for 65% (11/17), followed by HPA related anti GP, accounting for 35% (6/17). The majority of patients had a platelet antibody positive typing rate below 30%, accounting for 71.4% (10/14). 【Conclusions】 The positive rate of platelet antibody of patients in Zhongshan area is significantly higher than that of voluntary blood donors, and most of them are anti-HLA Ⅰ and anti-GP, and the incidence of anti-CD36 is extremely low. Therefore, it is necessary to establish a known platelet antigen donor bank, and at the same time, carry out platelet antibody testing and matching of patients, which is helpful to solve the issue of platelet transfusion refractoriness.

Objective:To explore the influence of age on the passive stiffness of women′s quadriceps muscles and the relationship between stiffness and muscle functioning.Methods:Twenty healthy elderly women formed the senior-aged group, while 20 young, healthy women were selected as the young group. The passive stiffness of the subjects′ vastus medialis, rectus femoris and vastus lateralis muscles was measured using ultrasound shear wave imaging with the knee flexed to 30°, 60°, 90° and 105°. Both groups completed the 6-minute walk test (6MWT) and the five times sit-to-stand test (FTSST). Maximum voluntary isometric contraction (MIVC) strength during knee extension was also measured.Results:The passive muscle stiffness of the rectus femoris and vastus lateralis muscles was significantly greater in the senior-aged group when the knee was flexed at 60°, 90° or 105°. The 6MWT and FTSST times of the young group were of course significantly better, and their MIVC at 60° and 90° of knee flexion was also significantly stronger. Rectus femoris stiffness among the senior-aged group was negatively correlated with MIVC when the knee was flexed at 60°.Conclusions:The passive stiffness of the muscles studied increases with age, especially in stretching. When the knee joint is flexed at 60° there tends to be a negative correlation between the MIVC of the quadriceps femoris and the stiffness of the rectus femoris among elderly females.

Objective:To develop an integrated point-of-care testing (POCT) reagent for genotying respiratory syncytial virus (RSV) and evaluate its performance.Methods:Specific primers and probes were designed based on the conserved sequences of the genomes of RSV A and B as well as ON1 and BA9 genotypes. The PCR reaction system and conditions were optimized. The vitrification technology of reagents and multiplex detection platform were integrated to develop the RSV genotyping POCT reagent. The sensitivity, specificity, reproducibility, and clinical performance of the product were then evaluated.Results:The sensitivity of the developed integrated RSV genotyping POCT reagent reached 500 copies/ml. It exhibited good specificity with no cross-reaction with clinically similar pathogens. The coefficient of variation of Ct values for both inter-batch and intra-batch reproducibility was less than 5%, indicating good reproducibility. In testing 53 clinical samples, the detection results showed high consistency and concordance with the reference reagent, with a positive concordance rate of up to 98.11%.Conclusions:The developed integrated RSV genotyping POCT reagent incorporates nucleic acid extraction, purification, and detection into a single process, achieving a "sample in, result out" workflow. It is simple to operate and provides accurate, reliable, and stable detection results. This product can be used for the genotyping of RSV A and B in POCT, offering support for the prevention, control, and diagnosis of RSV.

Objective To investigate value and risk factors of pelvic floor three-dimensional ultrasound parameters in single spontaneous delivery primipara with bladder neck funnel formation at 6-8weeks after delivery.Methods A total of 108 cases of single spontaneous delivery primipara were selected as research objects according to three-dimensional ultrasound indexes of pelvic floor,they were assigned into two groups:Observation group(40 cases)with bladder neck funnel formation and control group(68 cases)without bladder neck funnel formation.Three-dimensional ultrasound indexes of pelvic floor and clinical symptoms were compared between two groups.Results The incidence of stress urinary incontinence(SUI),anterior vaginal wall prolapse,bladder neck distance(BND),posterior urethrovesical angle(PUA)in Valsalva state,levator hiatus area(LHA)in resting and Valsalva states,levator hiatus length(LHL),and changes in LHL(△ LHL)in observation group were all higher than those in control group(P＜0.05).Logistic regression analysis revealed that SUI and anterior vaginal wall prolapse are independent risk factors for bladder neck funnel formation,adjusted OR were 11.255 and 3.643;Resting state and Valsalva state LHA,LHL,and △ LHL were also independent risk factors for bladder neck funnel formation(P＜0.05).Receiver operating characteristic(ROC)curve analysis revealed that area under the curve(AUC)for diagnosing bladder neck funnel formation in SUI and anterior vaginal wall prolapse were 0.744 and 0.615,respectively;In ultrasound indicators,when the LHL of Valsalva state is＞63.5mm,the AUC is 0.706;The diagnostic sensitivity of the combination factor 1(resting state LHA+Valsalva state LHA)is 95.0％;The diagnostic specificity of the combination factor 2(Valsalva state LHL+△LHL)is 80.9％.Conclusion In patients of single spontaneous delivery primipara with bladder neck funnel formation,occurrence of SUI and vaginal anterior wall prolapse,increased LHA and LHL under two states and △LHL were independent risk factors.The diagnostic sensitivity of the combination factor 1 was higher than single index,the diagnostic specificity of the combination factor 2 was higher.

Objective:To examine the burden and trends of acute viral hepatitis in Guangdong Province from 1990 to 2019, and provide reference evidences for hepatitis prevention and control in the province.Methods:Data on acute viral hepatitis (hepatitis A, B, C, and E) in Guangdong from 1990 to 2019 were extracted from the Global Burden of Disease Study 2019 database. The incidence, prevalence, mortality, and disability-adjusted life years (DALY) data were analyzed by age and gender, and the estimated annual percentage change (EAPC) was calculated to describe the changing trends in disease burden.Results:From 1999 to 2019, the standardized incidence, prevalence, mortality, and DALY of acute viral hepatitis in Guangdong were higher than the national averages. In 2019, 51.43% (2 245 087/4 365 221) of acute viral hepatitis cases in Guangdong Province were mainly attributed to hepatitis B, and 77.18% (106/138) of deaths were due to acute hepatitis B. In different age groups, except for acute hepatitis B, which was more common in adults, the incidence rates of other types of viral hepatitis such as hepatitis A, B, and E showed an overall decreasing trend with age. The mortality rates of different types of acute viral hepatitis, except for the <5 age group, increased with age. The overall incidence and mortality rates of acute viral hepatitis were higher in men than in women.Conclusions:The overall burden of acute viral hepatitis in Guangdong declined in 2019, but remained higher than the national level. Further efforts are needed to strengthen hepatitis prevention and screening in different population in Guangdong Province, especially in children and the elderly.

Objective:To analyze the effects of SARS-CoV-2 infection and vaccination on virus mutation.Methods:The whole genome sequencing sequences of 2 659 local SARS-CoV-2 specimens from Jiangsu Province in 2023 were selected for analysis, and relevant information such as demographic and clinical characteristics were collected, and the effects of infection and vaccination on the genome-wide mutation rate and S gene′s selective pressure of the virus were analyzed by univariate and multivariate linear regression models.Results:The average age of these infected patients was 55.0 (31.0, 74.0) years, 1 150 cases (43.2%) in the age group of ≥60 years, 1 367 cases (51.4%) were males, 2 044 cases (76.9%) had a history of COVID-19 vaccination, and 1 629 cases (61.3%) had the first-time infection. The clinical symptoms of the infected patients were mainly mild, with a total of 2434 cases (91.5%), and 29 cases (1.1%) with severe symptoms or more. The average substitution rate of SARS-CoV-2 was 9.69 (9.38, 9.98)×10 -4 subs/site/year, and the dN/dS value of the S gene was 6.08 (5.56, 8.66), which was significantly greater than that of 1 ( P<0.001), indicating positive selection. The result of univariate and multivariate linear regression model analysis showed that the SARS-CoV-2 substitution rate was higher in those with vaccination history and reinfection, aged 20-30 years, ≥60 years, and the SARS-CoV-2 substitution rate was lower in males with moderate clinical symptoms and severe disease and above. Those with a history of vaccination and reinfection, aged 50-60 years old, ≥60 years old have smaller S gene dN/dS. Conclusions:Under the immune pressure exerted by vaccination and infection, the genome-wide mutation of SARS-COV-2 accelerated, but the non-synonymous mutation rate of the S gene decreased. The mechanism causing these phenomena needs further study.