ObjectiveTo observe the clinical efficacy of Tangshen Dihuang decoction in treating diabetic kidney disease （DKD） with liver-kidney Yin deficiency and blood stasis syndrome and its impact on gut microbiota. MethodsA randomized controlled clinical trial was conducted， in which 102 DKD patients with liver-kidney Yin deficiency and blood stasis syndrome were randomly assigned to the Tangshen Dihuang decoction group and the control group. Each group consisted of 51 cases， and the treatment period was 3 months. The primary efficacy indicators included urinary albumin-to-creatinine ratio （UACR）， fasting blood glucose （FBG）， 2-hour postprandial blood glucose （2 hPBG）， glycated hemoglobin （HbA₁C）， serum creatinine （SCr）， blood urea nitrogen （BUN）， angiotensinⅡ （AngⅡ）， serum cystatin C （Cys-C）， urinary N-acetyl-β-D-glucosaminidase （NAG）， urinary β₂-microglobulin （Uβ₂-MB）， traditional Chinese medicine （TCM） symptom scores， and gut microbiota. ResultsAfter treatment， the total response rate in the Tangshen Dihuang decoction was 87.23% （41/47）， which was higher than that （69.57%， 32/46） in the control group （Z=4.30， P<0.05）. After treatment， the TCM symptom scores decreased in both groups （P<0.01） and were lower in the Tangshen Dihuang decoction group than in the control group （P<0.01）. After treatment， the control group showed decreases in UACR， Uβ₂-MG， AngⅡ， and FBG （P<0.05） as well as 2 hPBG and HbA₁C （P<0.01）， and no significant differences in BUN， Cys-C， eGFR， SCr， and NAG. The Tangshen Dihuang decoction group showed increased eGFR （P<0.05）， declined levels of UACR， BUN， Cys-C， Uβ₂-MB， AngⅡ， FBG， 2 hPBG， NAG， and HbA₁C （P<0.01）， and no significant difference in SCr. The Tangshen Dihuang decoction group had lower BUN （P<0.05）， Cys-C （P<0.05）， AngⅡ （P<0.05）， 2 hPBG （P<0.05）， Uβ₂-MG （P<0.01）， and NAG （P<0.01） and higher eGFR level （P<0.05） than the control group. After treatment， the control group showed declines in Shannon， Observed_species， and Chao1 indices （P<0.05）. The samples from both groups showed statistically significant differences in the principal coordinates analysis （PCoA） plot based on Anosim analysis （P<0.05）. After treatment， the Tangshen Dihuang decoction group showed decreased relative abundance of Actinobacteria （P<0.05）. Moreover， the relative abundance of Actinobacteria was significantly lower in the Tangshen Dihuang decoction group than in the control group （P<0.05）. At the genus level， the control group showed decreased relative abundance of Bifidobacterium （P<0.05）， and the Tangshen Dihuang decoction group presented increased relative abundance of Bifidobacterium and Blautia_A （P<0.05）. After treatment， the Tangshen Dihuang decoction group had higher relative abundance of Bifidobacterium than the control group （P<0.01）. ConclusionTangshen Dihuang decoction has a significant therapeutic effect on DKD patients with liver-kidney Yin deficiency and blood stasis syndrome. It can markedly relieve clinical symptoms and reduce proteinuria and postprandial blood glucose by antagonizing the local renin-angiotensin system （RAS） and alleviating gut microbiota dysbiosis.

ObjectiveTo compare the mosquito capture efficacy of the BG-trap method and the light trap method, and to explore the precisely applicable scenarios for the two monitoring approaches. MethodsFrom May to October 2024, one BG-trap and one light trap were placed at eight monitoring sites, with a distance of over 10 meters between the two traps at each site. Surveillance was performed twice monthly to compare the two surveillance methods in terms of mosquito species composition, the proportional distribution of female mosquitoes across different habitats, and seasonal fluctuations in mosquito density. ResultsThere were statistically significant differences in mosquito species composition (χ²=60.61, P<0.001) and female mosquito species composition (χ²=37.12, P<0.001) between the BG-trap method and the light trap method, with the BG-trap method capturing a higher proportion of Aedes albopictus than the light trap method. The average density of mosquitoes monitored by the BG-trap method [13.36 mosquitoes·(trap·night)^-1] was higher than that monitored by the light trap method[4.70 mosquitoes·(trap·night)^-1]. There was statistically significant difference (t=6.70, P<0.001) for the mosquito density between the two methods, and there was a significant positive correlation (r=0.909, P<0.001) for the mosquito density indices between the two methods. The average density of female mosquitoes monitored by the BG-trap method [8.18 mosquitoes·(trap·night)^-1] was higher than that monitored by light trap method [3.36 mosquitoes·(trap·night)^-1]. There was statistically significant difference (t=6.00, P<0.001) for the female mosquito density between the two methods, and there was a significant positive correlation (r=0.929, P<0.001) for the female mosquito density indices between the two methods. In the four different habitats, the constituent ratio of mosquitoes and female mosquitoes monitored by the BG-trap method in descending order were urban residential area (43.80% and 40.25%), rural farm households (35.55% and 38.60%), hospital (12.70% and 12.87%), and park (7.95% and 8.28%), while the constituent ratio of mosquitoes and female mosquitoes monitored by the light trap method in descending order were rural farm households (34.37% and 43.34%), hospital (33.26% and 30.03%), park (17.96% and 15.17%), and urban residential area (14.41% and 11.46%). In different habitats, there were statistically significant differences for mosquito composition ratios (χ²=189.64, P<0.001) and female mosquito composition ratios (χ²=109.15, P<0.001) between the two monitoring methods. ConclusionThe BG-trap method offers advantages such as simple operation, compact equipment, and high efficiency, with a higher trapping rate for Aedes albopictus compared to the light trap method. In routine monitoring, the BG-trap method can serve as a supplement to the light trap method, while for emergency monitoring of Aedes albopictus in dengue fever outbreak areas and mosquito collection efforts, the BG-trap method is more effective than the light trap method.

OBJECTIVES: To investigate the impact of prenatal fear stress on placental amino acid transport and emotion and cognition development in offspring rats. METHODS: Thirty pregnant Wistar rats were randomized equally into control and fear stress (induced using an observational foot shock model) groups. In each group, placental and serum samples were collected from 6 dams on gestational day 20, and the remaining rats delivered naturally and the offspring rats were raised under the same conditions until 8 weeks of age. Emotional and cognitive outcomes of the offspring rats were assessed with behavioral tests, and placental structure was examined using HE staining. Bioinformatics analysis was used to identify differentially expressed placental transporter genes under fear stress. The expressions of system A and system L amino acid transporters, along with other specialized transporters, were detected using qRT-PCR and Western blotting. Fetal serum amino acid concentrations were determined by HPLC. The correlations between fetal amino acid levels and behavioral outcomes of the offspring rats were analyzed. RESULTS: The dams with fear stress showed reduced open-field activity and increased freezing behavior with significantly decreased placental weight, fetal weight, and fetal-to-placental ratio. Bioinformatics analysis revealed 28 differentially expressed transporter genes involved mainly in amino acid transport. In the fear stress group, fetal serum amino acid levels were significantly lowered and Slc38a1, Slc43a1, Slc43a2, Slc7a8, Slc6a6, Slc1a1 and Slc6a9 mRNA and protein expressions were all downregulated. The offspring rats in fear stress group exhibited decreased novel object preference and spontaneous alternation with reduced open arm exploration and increased immobility in emotional tests. Lower early-life amino acid levels was found to correlate with impaired adult cognition. CONCLUSIONS Prenatal fear stress in rats impairs placental amino acid transporter expression and reduces fetal serum amino acid levels, potentially contributing to long-term cognitive deficits in the offspring rats.
Animals ; Female ; Pregnancy ; Placenta/metabolism* ; Fear ; Rats ; Rats, Wistar ; Cognition ; Prenatal Exposure Delayed Effects ; Stress, Psychological ; Amino Acids/blood* ; Amino Acid Transport Systems/metabolism*

OBJECTIVE To screen and obtain nanobodies with neutralizing activity against granulo-cyte-macrophage colony stimulating factor(GM-CSF)to contribute to investigations into the mecha-nism of inflammation interventions and the development new drugs.METHODS Recombinant human GM-CSF was subcutaneously injected to immunize camels.Peripheral blood was collected after five immunizations,and mononuclear cells were isolated.Total mRNA was extracted,and the variable domains of the heavy chain of heavy-chain antibody(VHH,also called nanobody)genes were obtained by PCR amplification after reverse transcription.The genes were cloned into the pADSCFV-S phage display vector and electrotransformed into TG1 competent cells to construct a nanobody immune library that was screened with recombinant human GM-CSF immobilized on a solid phase.The VHH genes specifi-cally binding to human GM-CSF were cloned into the pABG eukaryotic expression vector before VHH-Fc samples were prepared by using the human embryonic kidney 293 fibroblast expression system.The binding activity of candidate VHH-Fc molecules to GM-CSF was investigated through ELISA response curves,and binding colorimetric values with different antigens were detected to determine their specificity.The binding affinity between VHH-Fc candidates and GM-CSF was measured using biolayer interferom-etry(BLI).The inhibition rate curve of VHH-Fc candidates on GM-CSF was detected through cell prolif-eration assays to determine its neutralization activity.The Uncle system was used to investigate its thermal stability.100 μg of VHH-Fc was injected into mice via the tail vein,and the serum concentration of VHH-Fc was quantitatively detected by ELISA to examine its pharmacokinetic curve in mice.RESULTS The camel serum titer of anti-human GM-CSF antibodies was higher than 1:800 000 after the fifth immuni-zation,and the capacity of the constructed nanobody library was about 5.55×107.Following the screening process,five candidate VHH-Fc molecules specifically binding to human GM-CSF were obtained.Among these,22N10 effectively neutralized the cell proliferation-promoting activity of GM-CSF(the IC50 value was 17.23 nmol·L-1).Subsequent studies revealed that 22N10 interacted with human GM-CSF with an affinity of 1.97×10-8 mol·L-1,blocked the binding of GM-CSF to its receptor CSF2Rα,exhibited good thermal stability(Tm1=59.2℃),and showed favorable metabolic characteristics in mice.CONCLU-SION A new candidate nanobody molecule 22N10 targeting human GM-CSF is obtained which is expected to facilitate the drug development and mechanism investigations.

In this study,the carboxylated magnetic beads were coupled with bivalent nanobodies a-gainst porcine epidemic diarrhea virus(PEDV)M protein to construct immunomagnetic beads(IM-NBs-Ⅱ),The capture and enrichment function of IMNBs-Ⅱ was verified by using PEDV propaga-ted in Vero cells.A one-step RT-qPCR detection method for PEDV was established by combining the characteristics of IMNBs-Ⅱ with the detection advantages of reverse transcription fluorescence quantitative polymerase chain reaction(RT-qPCR).Specific analysis found that this method has no cross reactivity with swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,porcine circovirus,indicating that it has good specificity.Sensitivity analysis re-vealed that the detection sensitivity of the RT-qPCR based on IMNBs-Ⅱ was increased 10 times compared to traditional RT-qPCR methods.Detection of the clinical samples confirm that the RT qPCR method based on IMNBs-Ⅱ is suitable for rapid and accurate detection of clinical feces and tissue samples.The method established in this study effectively avoids contamination issues during nucleic acid extraction,simplifies experimental procedures,and saves detection time,which pro-vides a method for efficient detection of PEDV.

Background::Atopic dermatitis (AD) is a chronic inflammatory skin disorder impacting populations worldwide, although its clinical characteristics and patient demographics remain uncharacterized in China. The aim of this study was to investigate the demographics, comorbidities, aggravating factors, and treatments in AD patients across different age groups in China.Methods::This cross-sectional study included Chinese AD patients from 205 hospitals spanning 30 provinces. Patients completed dermatologist-led surveys of general medical history, comorbidities, AD-related aggravating factors, and medications. Two-level mixed-ordered logistic regression was used to evaluate aggravating factors.Results::Overall, 16,838 respondents were included in the final analysis (aged 30.9 ± 24.1 years). The proportion of severe AD was the highest in patients with AD onset at ≥60 years (26.73%). Allergic rhinitis and hypertension were the most common atopic and metabolism-related non-atopic comorbidities, respectively. AD severity was significantly associated with chronic urticaria, food allergies, and diabetes. Aggravating factors including foods, seasonal changes, and psychological factors were also linked to AD severity. The cross-sectional survey implied that severe AD may be related to the undertreatment of effective systemic or topical interventions.Conclusion::To enhance the management of AD, it is crucial to consider both aggravating factors and the increased utilization of systemic immunotherapy.Registration::ClinicalTrials.gov, NCT05316805

Objective:To explore the impacts of different frequencies of orthogonal image-guided verification on set-up error correction and the reference values of planning target volume (PTV) margins in postoperative radiotherapy for breast cancer, in order to provide recommendations of the verification frequency in clinical practice.Methods:A total of 80 breast cancer patients who received postoperative intensity-modulated radiotherapy for breast cancer at the Affiliated Union Hospital of Fujian Medical University from January 2021 to January 2022 were enrolled. Orthogonal image-guided verification was conducted before each radiation treatment to determine initial and residual set-up errors. The error data were statistically analyzed by categorizing patients into groups A, B, C, D, E, and F, based on assumed verification frequencies (daily, every 2, 3, 4, and 5 d, and no correction). The magnitude, distribution, and differences of set-up errors of various groups were analyzed, and the PTV margins were calculated using a reference formula.Results:The collected initial set-up errors of the 80 patients were (3.49±3.43), (2.87±2.88), and (2.75±2.35) mm, in x, y, and z directions, respectively. The residual set-up errors decreased gradually with increasing verification frequency. In the case of daily image-guided verification, the residual set-up errors in x, y, and z directions decreased to (1.63±0.90), (1.63±0.93), and (1.55±0.90)mm, respectively. The reference values of PTV margins calculated under different verification frequencies decreased with an increase in the verification frequency. Under daily verification, the reference values of PTV margins calculated based on set-up errors decreased from initial 6.60, 6.29, and 4.22 mm (without correction) to 1.27, 1.37, and 1.28 mm, respectively. Differences in the reference values under verification frequencies every 3, 4, and 5 d were less than 0.81 mm. Conclusions:Daily image-guided verification (including correction) can effectively reduce set-up errors in image-guided radiotherapy for breast cancer. The set-up errors differ under varying verification frequencies, with a higher frequency corresponding to better correction effects and smaller PTV margins required.

Objective:To investigate the factors associated with prolonged QTc interval in long-term hospitalized patients with schizophrenia, and to develop and validate a predictive model.Methods:A cross-sectional study was conducted involving 215 long-term hospitalized patients with schizophrenia at Xianyang Hospital, Yan'an University from January to December 2023. The incidence of prolonged QTc intervals among the patients was analyzed. Univariate and multivariate analyses of the factors associated with prolonged QTc intervals were performed. A mathematical model was developed to predict prolonged QTc intervals in long-term hospitalized patients with schizophrenia, and the predictive performance of the model was evaluated.Results:Among the 215 patients, 42 exhibited prolonged QTc intervals, with an incidence rate of 19.53%. Univariate analysis revealed statistically significant differences between the groups with and without prolonged QTc in terms of age, gender, body mass index, categories of antipsychotic medication used, fasting blood glucose, and potassium levels ( t = -5.66, χ2 = 29.03, t = -2.01, χ2 = 10.96, t = -5.78, t = 5.92, all P < 0.05). Multivariate logistic regression analysis indicated that age, gender, fasting blood glucose, and potassium levels were independent factors affecting QTc interval prolongation ( OR = 1.097, 10.221, 2.449, 0.014, all P < 0.05). The goodness-of-fit of the logistic regression model was validated to be satisfactory (Hosmer-Lemeshow χ2 = 14.56, P > 0.05). A risk nomogram model based on the variables from the multivariate analysis had a C-index of 0.713. Receiver operating characteristic curve analysis was performed using the independent influential factors and their P values from the logistic regression model to predict the probability of QTc interval prolongation. The area under the curve values were 0.762, 0.725, 0.730, 0.792, and 0.920, respectively. Conclusions:The incidence of prolonged QTc interval is relatively high in long-term hospitalized patients with schizophrenia. Female gender, older age, hyperglycemia, and hypokalemia are all associated with prolonged QTc intervals. The mathematical model developed based on these factors demonstrates good predictive performance for QTc interval prolongation.

Objective To investigate the risk factors of secondary traumatic coagulopathy in patients with traffic trauma.Methods A total of 122 patients with traffic trauma in Department of Emergency Surgery,the First Affiliated Hospital of Anhui Medical University(Dongcheng Hospital)from April 2022 to March 2023 were retrospectively collected.Patients were divided into the occurrence group(n=30)and the non-occurrence group(n=92).Body temperature,acidosis,hematoma enlargement after head CT examination,Glasgow coma score(GCS),trauma severity score(ISS),platelet count(PLT)and hemoglobin(Hb)at admission were collected and compared between the two groups.Multivariate Logistic regression was used to analyze risk factors of secondary traumatic coagulopathy in traffic trauma patients,and receiver operating characteristic(ROC)curve was used to analyze the predictive value of related factors for secondary traumatic coagulopathy.Results The body temperature,GCS,PLT and Hb in the occurrence group were lower than those in the non-occurrence group,while the ISS,incidence of acidosis and the incidence of increased hematoma after CT examination were higher than those in the non-occurrence group(all P<0.05).Multivariate Logistic regression analysis showed that low GCS,low Hb level,high ISS,acidosis and increased hematoma after head CT examination were risk factors at admission(P<0.05).The area under ROC curve(AUC)of the five factors combined for secondary traumatic coagulopathy in traffic trauma patients was 0.963,sensitivity was 83.3%and specificity was 96.7%.Conclusion Decreased GCS and Hb on admission,high ISS,acidosis and increased hematoma during head CT review are risk factors for secondary traumatic cotting disorder in traffic trauma patients,and the combination of these factors has high predictive value.

In this study,the carboxylated magnetic beads were coupled with bivalent nanobodies a-gainst porcine epidemic diarrhea virus(PEDV)M protein to construct immunomagnetic beads(IM-NBs-Ⅱ),The capture and enrichment function of IMNBs-Ⅱ was verified by using PEDV propaga-ted in Vero cells.A one-step RT-qPCR detection method for PEDV was established by combining the characteristics of IMNBs-Ⅱ with the detection advantages of reverse transcription fluorescence quantitative polymerase chain reaction(RT-qPCR).Specific analysis found that this method has no cross reactivity with swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,porcine circovirus,indicating that it has good specificity.Sensitivity analysis re-vealed that the detection sensitivity of the RT-qPCR based on IMNBs-Ⅱ was increased 10 times compared to traditional RT-qPCR methods.Detection of the clinical samples confirm that the RT qPCR method based on IMNBs-Ⅱ is suitable for rapid and accurate detection of clinical feces and tissue samples.The method established in this study effectively avoids contamination issues during nucleic acid extraction,simplifies experimental procedures,and saves detection time,which pro-vides a method for efficient detection of PEDV.