ObjectiveTo explore the possible mechanisms of Wenyang Huazhuo Formula （温阳化浊方， WHF） in improving reproductive aging from the perspective of the ovarian mechanical microenvironment. MethodsThe experiment included five groups， 3-month group （20 female mice at 3 months of age）， 6-month group （20 female mice at 6 months of age）， 6-month + WHF group （20 female mice at 5 months of age treated with WHF）， 9-month group （20 female mice at 9 months of age）， and 9-month + WHF group （20 female mice at 8 months of age treated with WHF）. The 6-month + WHF group and 9-month + WHF group were orally administered WHF 41.2 g/（kg·d） once daily for 4 consecutive weeks. The other three groups received no intervention. Reproductive hormone levels were measured by ELISA. HE staining was used to count the numbers of various stages of follicles. Ovarian hyaluronic acid （HA） content and collagen fiber content were measured to evaluate the ovarian mechanical microenvironment. Superovulation was performed to observe the number of eggs obtained， as well as the number of offspring and birth weight to assess fertility. The in vitro fertilization and blastocyst culture of oocytes from female offspring in each group were observed to evaluate the effect of WHF on offspring reproductive potential. ResultsCompared with the 3-month group， the 6-month group and 9-month group showed significantly decreased serum levels of gonadotropin-releasing hormone （GnRH）， follicle-stimulating hormone （FSH）， and luteinizing hormone （LH）， decreased ovarian collagen content， and reduced numbers of primordial and secondary follicles. In contrast， the numbers of primary follicles， antral follicles， and atretic follicles increased. The levels of anti-Müllerian hormone （AMH）， ovarian HA content， and the fertilization rate， cleavage rate， and blastocyst formation rate of oocytes from offspring were significantly lower （P＜0.05）. Compared with the 6-month group， the 6-month + WHF group showed significantly reduced serum levels of GnRH， FSH， and LH， with a significant decrease in primary follicles， antral follicles， and atretic follicles as well as increase of AMH levels， ovarian HA content， number of primordial and secondary follicle， egg count， and offspring birth weight （P＜0.05）. Compared with the 9-month group， the 9-month + WHF group exhibited reduced GnRH， FSH， and collagen fiber content， as well as reduced number of primary follicles， antral follicles， and atretic follicles. However， AMH levels， ovarian HA content， number of primordial and secondary follicle， egg count， offspring numbers， birth weight， fertilization rate， cleavage rate， and blastocyst formation rate of oocytes from offspring all significantly increased （P＜0.05）. ConclusionWHF can significantly improve the ovarian reserve， fertility， and reproductive potential in offspring during reproductive mid-life and late-life stages. Its effect may be related to the remodeling of the mechanical microenvironment of aging ovaries. Moreover， the effect on the mechanical microenvironment remodeling of late-stage ovaries and the improvement of the offspring reproductive potential is more significant.

Objective To analyze the operation of Measles Surveillance System (MSS) in Hebei Province, and to provide evidence for measles elimination. Methods Measles surveillance data was collected from the MSS from 2020 to 2023, and a modified weighted technology for order preference by similarity to an ideal solution (TOPSIS) method was used to evaluate the surveillance indexes of measles in Hebei Province. Results The operation quality of the measles surveillance system in Hebei Province was improved year by year, with the highest quality in 2023, and all the indicators reached the monitoring program standards. The quality of measles surveillance system was not balanced among cities, and the main influencing factor was the substandard sensitivity indicators. The quality of measles surveillance system was the highest in Baoding City and the lowest in Zhangjiakou City. Conclusion The measles surveillance system in Hebei province is running well, and the sensitivity of the surveillance system should be improved to keep the high-quality operation of the surveillance system.

AIM: To evaluate the safety and efficacy of allogeneic intrastromal lenticule implantation combined with corneal collagen cross-linking(CXL)in patients with moderate to advanced keratoconus.METHODS: A retrospective case series analysis was conducted. A total of 19 patients(20 eyes)with moderate to advanced keratoconus who underwent combined allogeneic intrastromal lenticule implantation and CXL at the Jinan Mingshui Eye Hospital from June 2021 to December 2023 were included. The uncorrected distance visual acuity(UCVA), thinnest corneal thickness, central corneal epithelial thickness, anterior corneal flat keratometry(Kf), steep keratometry(Ks), and mean keratometry(Km), as well as the first applanation time(A1T), the first applanation length(A1L), the velocity during the first applanation moment(VIN), the second applanation time(A2T), the second applanation length(A2L), the velocity during the second applanation moment(VOUT), highest concavity time(HCT), highest concavity radius(HCR), peak distance(PD), deformation amplitude(DA), stiffness parameter at first applanation(SP-A1), integrated radius(IR), central corneal thickness(CCT), intraocular pressure(IOP), corneal thickness-corrected IOP, biomechanically intraocular pressure IOP(bIOP), and corneal thickness variation rate(ARTH)were compared between the two groups before surgery and at 1 wk, 1, 3 and 6 mo after surgery.RESULTS: All patients successfully completed the surgery without any intraoperative complications. No significant differences were observed between pre-operative and post-operative measurements for UCVA or the corneal biomechanical parameters, including A1L, A2L, PD, A1T, A2T, VIN, VOUT, DA, IOP, and bIOP(all P>0.05). Significant differences were found between pre-operative and post-operative values for corneal thinnest point thickness, central corneal epithelial thickness, Kf, Ks, Km, and the corneal biomechanical parameters, including HCT, HCR, SP-A1, ARTH, IR, and CCT(all P<0.05). The anterior corneal curvature demonstrated an initial increase followed by a decrease post-operatively. Furthermore, significant differences were observed between pre-operative and post-operative values for HCT, HCR, SP-A1, ARTH, IR, and CCT(all P<0.005).CONCLUSION: Allogenic intrastromal lenticule implantation combined with corneal collagen cross-linking demonstrates favorable safety and stability in treating moderate-to-advanced keratoconus. This combined procedure effectively increases corneal thickness and rigidity, resulting in corneas that are more resistant to deformation postoperatively.

This study aims to establish a high-performance size-exclusion chromatography (HPSEC) method for determining the content of the classical swine fever virus (CSFV) E2 protein and screen the optimal stabilizer to enhance the stability of this protein. The optimal detection conditions were determined by optimizing the composition of the mobile phase, and characteristic chromatographic peaks were identified by SDS-PAGE and Western blotting. The specificity, repeatability, precision, linearity, limit of detection (LOD), and limit of quantitation (LOQ) of the method were assessed. The method established was used to determine the content of CSFV E2 protein antigen and vaccine. Differential scanning fluorimetry (DSF) was employed to screen the buffer system, pH, and salt ion concentrations, and sugar, amino acid, and alcohol stabilizers were further screened. The results showed that using a 200 mmol/L phosphate buffer provided the best column efficiency. An antigen-specific chromatographic peak appeared at the retention time of 18 min, which was identified as the CSFV E2 protein by SDS-PAGE and Western blotting. The method exhibited high specificity for detecting the CSFV E2 protein, with no absorbance peak observed in the blank control. The relative standard deviation (RSD) of the peak area for six repeated injections of the CSFV E2 protein was 0.74%, indicating good repeatability of the method. The RSD for repeated detection of two different concentrations of CSFV E2 protein samples by different operators at different time points was less than 2%, suggesting good intermediate precision of the method. The peak area of the CSFV E2 protein was linearly related to its concentration, with the regression equation showing R² of 1.000. The LOD and LOQ of the method were 14.88 μg/mL and 29.75 μg/mL, respectively. Application of the developed method in the detection of three batches of CSFV E2 protein antigen and three batches of vaccine demonstrated results consistent with those from the bicinchoninic acid (BCA) assay, which meant that the method could accurately determine the content of CSFV E2 protein antigen and vaccine. The DSF method identified 50 mmol/L Tris-HCl at pH 8.0 as the optimal buffer, and the addition of sugar and alcohol stabilizers further improved the stability of the CSFV E2 protein. The HPSEC method established in this study is simple, fast, and exhibits good accuracy and repeatability, enabling precise measurement of the CSFV E2 protein content. It is expected to play a crucial role in the quality control of the CSFV E2 vaccine. Furthermore, the strategy for improving the CSFV E2 protein stability, identified through DSF screening, has significant implications for enhancing the stability of the CSFV E2 vaccine.
Classical Swine Fever Virus/chemistry* ; Chromatography, Gel/methods* ; Animals ; Swine ; Viral Envelope Proteins/immunology*

Risk assessment models for periodontal disease provide dentists with a precise and consolidated evaluation of the prognosis of periodontitis, enabling the formulation of personalized treatment plans. Periodontal risk assessment systems have been widely applied in clinical practice and research. The application fields of periodontal risk assessment systems vary based on the distinctions between clinical periodontal parameters and risk factors. The assessment models listed below are commonly used in clinical practice, including the periodontal risk calculator (PRC), which is an individual-based periodontal risk assessment tool that collects both periodontal and systemic information for prediction; the periodontal assessment tool (PAT), which allows for quantitative differentiation of stages of periodontal disease; the periodontal risk assessment (PRA) and modified periodontal risk assessment (mPRA), which are easy to use; and the classification and regression trees (CART), which assess the periodontal prognosis based on a single affected tooth. Additionally, there are orthodontic-periodontal combined risk assessment systems and implant periapical risk assessment systems tailored for patients needing multidisciplinary treatment. This review focuses on the current application status of periodontal risk assessment systems.

ObjectiveTo investigate the mechanism of salvianolic acid F （Sal F） in repairing the high glucose-induced injury in human kidney-2 （HK-2） cells via the B-cell lymphoma-2 （Bcl-2）-associated X protein （Bax）/cysteinyl aspartate-specific proteinase 3 （Caspase-3）/gasdermin-E （GSDME） pathway. MethodThe cell counting kit-8 （CCK-8） was used to measure the relative viability of HK-2 cells exposed to high glucose and different concentrations （2.5， 5， 10， 20 μmol·L^-1） of Sal F and the relative viability of HK-2 cells treated with Sal F for different time periods. The levels of lactate dehydrogenase （LDH） and interleukin-1β （IL-1β） in the supernatant of the cell culture were measured by the LDH assay kit and enzyme-linked immunosorbent assay （ELISA） kit， respectively. Flow cytometry combined with Annexin V-FITC/propidium iodide （PI） and Hoechst 33342/PI staining was employed to reveal the proportion of PI-positive HK-2 cells exposed to high glucose. Western blotting was employed to determine the protein levels of Bax， Bcl-2， cytochrome C， cysteinyl aspartate-specific proteinase （Caspase）-9， Caspase-3， and GSDME in the HK-2 cells exposed to high glucose and treated with Sal F. The 2，7-dichlorodihydrofluorescein diacetate fluorescence probe （DCFH-DA） and mitochondrial membrane potential assay kit （JC-1） were used to determine the production of reactive oxygen species （ROS） and the mitochondrial membrane potential in the HK-2 cells exposed to high glucose and treated with Sal F. ResultCompared with the blank group， the model group showed decreased cell viability （P<0.01）， elevated levels LDH and IL-1β， increased proportion of PI-positive cells （P<0.01）， up-regulated protein levels of Bax， cytochrome C， Caspase-9， Caspase-3， and GSDME （P<0.01）， down-regulated protein level of Bcl-2 （P<0.01）， decreased mitochondrial membrane potential， and excessive ROS accumulation. Compared with the model group， Sal F repaired the high glucose-induced injury in HK-2 cells （P<0.05）， lowered the levels of LDH and IL-1β （P<0.05， P<0.01）， and decreased the proportion of PI-positive cells （P<0.01）. In addition， Sal F down-regulated the protein levels of Bax， cytochrome C， Caspase-9， Caspase-3， and GSDME and up-regulated the protein level of Bcl-2 （P<0.05， P<0.01）， increased the mitochondrial membrane potential， and decreased the accumulation of ROS in HK-2 cells. ConclusionSal F can reduce the production of ROS， restore the balance of mitochondrial membrane potential， and inhibit pyroptosis via the Bax/Caspase-3/GSDME signaling pathway to repair the high glucose-induced injury in HK-2 cells.

Objective To investigate the mechanism of astragaloside Ⅰ,the active constituent of milkvetch root,in inhibiting podocyte injury and improving diabetic kidney disease.Methods According to the body weight,60 male db/db mice were randomly divided into the model group,astragaloside Ⅰ low-dose group(10 mg/kg),astragaloside Ⅰ medium-dose group(20 mg/kg),astragaloside Ⅰ high-dose group(40 mg/kg),and valsartan group(10mg/kg),with 12 mice per group.Twelve db/db littermate control db/m mice were used as the control group.The drug was administered by gavage for 8 weeks.Transmission electron microscope was used to observe the ultrastructure of the kidney;immunohistochemistry and Western blotting were used to detect the expression of nephrotic protein(nephrin),a marker of renal podocytes;enzyme-linked immunosorbent assay was used to detect the contents of interleukin-1β(IL-1β)and interleukin-18(IL-18)in the serum of mice;Western blotting was used to detect the protein expressions of NOD-like receptor thermoprotein domain-related protein 3(NLRP3),cysteinyl aspartate specific proteinase 1(Caspase-1),and Gasdermin D(GSDMD)in kidney tissue.Results Compared with the control group,the glomeruli of the model group showed obvious podocyte loss and foot process fusion;the protein expression of nephrin was decreased(P＜0.05);the contents of IL-1 β and IL-18 in serum were increased(P＜0.05);the protein expressions of NLRP3,Cleaved-Caspase-1,and GSDMD-N were increased(P＜0.05).Compared with the model group,the renal pathological damage in the astragaloside Ⅰ administration groups were alleviated;the protein expression of nephrin was increased(P＜0.05);the contents of IL-1β and IL-18 in serum were decreased(P＜0.05);the protein expressions of NLRP3,Cleaved-Caspase-1,and GSDMD-N were decreased(P＜0.05).Conclusion Astragaloside Ⅰ may play a role in intervening diabetic kidney disease by inhibiting pyroptosis and improving podocyte injury.

Objective:To investigate the influencing factors of cardiac autonomic dysfunction in maintenance hemodialysis (MHD) patients by recording 48 h heart rate variability.Methods:It was a single-center cross-sectional study. MHD patients at the Hemodialysis Center of Peking University People's Hospital between October 1, 2021 and December 31, 2022 were enrolled in the study. These patients initiated hemodialysis for more than three months and were older than 18 years old, and patients with tachyarrhythmia, implanted cardiac pacemaker and the recording time less than 48 h were excluded. Demographic data, comorbidity, laboratory data, hemodialysis session data and heart rate variability were collected. Multivariate linear regression model was used to analyze the influencing factors for cardiac autonomic dysfunction in MHD patients.Results:A total of 110 patients were enrolled in the study, including 37 females (33.6%) and 36 diabetic patients (32.7%). The age of the patients was (57.8±14.8) years old, and the median dialysis vintage was 73.00(27.75±130.25) months. At baseline, the serum phosphate level was (1.6±0.4) mmol/L, and the N-terminal pro B-type natriuretic peptide (NT-proBNP) after ln transformed {ln[NT-proBNP(ng/L)]} was 8.4±1.2. The standard deviation of all normal R-R interval (SDNN) was (90.6±27.9) ms, ln[root mean square of successive differences in R-R interval (RMSSD, ms)] 3.2±0.8, ln[low frequency (ms 2)] 3.4±1.3, ln[high frequency (ms 2)] 3.1±1.4, and ln[low frequency/high frequency ratio] 0.28±0.64. After adjusting the age, coronary heart disease, diabetes, hemoglobin, serum phosphate and 25-hydroxy-vitamin D, serum natrium ( β=2.042, 95% CI 0.021–4.064, P=0.048) and ln[NT-proBNP (ng/L)] ( β=-7.027, 95% CI -12.247–-1.808, P=0.009) were independently correlated with SDNN (adjusted R2=0.218). Univariate linear regression model showed that diabetes was correlated with ln[low frequency(ms 2)] of MHD patients ( β=-0.659, 95% CI -1.171–-0.146, P=0.012), but in the multivariate linear regression model, significant correlation between diabetes and low frequency was not found. After adjusting the diabetes, coronary heart disease, dialysis vintage, hemoglobin, serum phosphate, serum albumin, pre-dialysis systolic blood pressure, post-dialysis systolic blood pressure, pre-dialysis diastolic blood pressure, increasing age ( β=-0.011, 95% CI -0.019–-0.003, P=0.007) and ln[NT-proBNP(ng/L)] ( β=-0.151, 95% CI -0.253–-0.048, P=0.004) were independently correlated with a decrease in the ln[low frequency/high frequency ratio]. In the multivariate linear regression model with ln[high frequency(ms 2)] or ln[RMSSD(ms)] as dependent variable, after adjusting the relevant factors, serum phosphate level was independently correlated with ln[RMSSD(ms)] ( β=-0.421, 95% CI -0.777–-0.065, P=0.021) or ln[high frequency(ms 2)] ( β=-0.752, 95% CI -1.325–-0.180, P=0.010). Conclusions:Hyperphosphatemia is an independent influencing factor of parasympathetic nervous system in MHD patients. Higher NT-proBNP is associated with lower SDNN and lower ratio of low frequency/high frequency, so serum phosphate control and volume control should be highlighted. Age is associated with autonomic dysfunction in MHD patients, so more attention should be paid to elder patients.

Objective:To investigate the effects of marathon exercise on knee cartilage volume and T2 relaxation time (T2 value) based on MRI.Methods:From December 2018 to December 2021, 25 healthy volunteers without long-distance running habits and 32 non-professional marathon runners with long-term long-distance running were recruited to undergo knee MRI 3D water-selective excitation (three dimensional water-selective excitation, 3D-WATS) and T2 mapping imaging were performed, and the cartilage volumes in 5 knee areas and T2 values in 42 subareas were extracted for analysis. To compare the cartilage volume and its ratio to body surface area of knee joint of healthy volunteers and non-professional marathon runners, the T2 value of cartilage in each subregion, and the correlation between marathon exercise intensity and the volume and T2 value of cartilage in different regions.Results:Compared with healthy volunteers, there was no significant difference in cartilage volume or the ratio of body surface area to body volume of non-professional marathon runners ( P>0.05). There were significant differences between healthy volunteers and non-professional marathon runners in cartilage T2 values of the median layer of medial condyle of femur (47.61±5.65 ms and 44.29±6.10 ms) and the deep layer of medial condyle of femur (36.82±9.05 ms and 31.67±7.59 ms), deep precondylar area of medial femur (38.37±4.68 ms and 34.09±4.19 ms), shallow area of medial condylar area of femur (52.17±11.11 ms and 45.51±7.76 ms), middle layer of medial condylar area of femur (49.09±5.08 ms and 45.63±5.04 ms), medial layer of anterior condylar region of lateral femur (45.69±4.68 ms and 42.57±5.77 ms), superficial layer of posterior condylar region of lateral femur (55.42±18.41 ms and 47.99±8.39 ms), deep layer of anterior tibial medial plateau (33.40±7.76 ms and 29.03±5.69 ms), deep layer of posterior tibial medial plateau (31.28±5.02 ms and 27.92±5.99 ms), deep layer of patellofemoral surface (35.65±6.99 ms and 32.30±5.28 ms), respectively ( P<0.05). In non-professional marathon runners, the medial tibial plateau cartilage volume was negatively correlated with step frequency ( r=-0.371, P=0.035), the lateral femoral condylar cartilage volume was negatively correlated with step frequency ( r=-0.365, P=0.043), and the lateral tibial plateau cartilage volume was negatively correlated with step frequency ( r=-0.550, P=0.001). The T2 value of the medial layer cartilage in the anterior tibial medial plateau region was negatively correlated with body weight ( r=-0.277, P=0.039) and body mass index ( r=-0.290, P=0.030). The T2 value of the superficial layer of patellofemoral surface was negatively correlated with the amount of running in 3 months ( r=-0.457, P=0.010). The superficial T2 value in the posterior lateral plateau of the tibia was negatively correlated with stride length ( r=-0.437, P=0.014), and the medial layer cartilage T2 value in the anterior condylar area of the lateral femur was negatively correlated with stride frequency ( r=-0.380, P=0.035). Conclusion:Marathon exercise had little effect on the knee cartilage volume, but had a certain effect on the cartilage T2 value, resulting in changes in cartilage structure. The higher the step frequency, the smaller the cartilage volume. The greater the body weight or body mass index, the greater the amount of running in 3 months, and the greater the stride length, the lower the cartilage T2 value.

Risk assessment models for periodontal disease provide dentists with a precise and consolidated evalua-tion of the prognosis of periodontitis,enabling the formulation of personalized treatment plans.Periodontal risk assess-ment systems have been widely applied in clinical practice and research.The application fields of periodontal risk assessment systems vary based on the distinctions between clinical periodontal parameters and risk factors.The assess-ment models listed below are commonly used in clinical practice,including the periodontal risk calculator(PRC),which is an individual-based periodontal risk assessment tool that collects both periodontal and systemic information for pre-diction;the periodontal assessment tool(PAT),which allows for quantitative differentiation of stages of periodontal dis-ease;the periodontal risk assessment(PRA)and modified periodontal risk assessment(mPRA),which are easy to use;and the classification and regression trees(CART),which assess the periodontal prognosis based on a single affected tooth.Additionally,there are orthodontic-periodontal combined risk assessment systems and implant periapical risk as-sessment systems tailored for patients needing multidisciplinary treatment.This review focuses on the current applica-tion status of periodontal risk assessment systems.