ObjectiveTo investigate the protective effects and mechanisms of Zhenzhu Tiaozhi capsules on the kidneys in the mouse model of diabetic kidney disease. MethodsThirty male C57BL/6J mice were selected as experimental objects. The model of diabetic kidney disease was induced by intraperitoneal injection of streptozotocin （STZ） at 40 mg·kg^-1 for 5 days combined with a high-fat diet （HFD）. Fasting blood glucose （FBG） ≥ 11.1 mmol·L^-1， increased urine volume， and continuous appearance of proteinuria indicated successful modeling. Mice were grouped as follows： Blank， model， low- and high-dose （0.98 and 1.96 g·kg^-1， respectively） Zhenzhu Tiaozhi capsules， and losartan potassium （30 mg·kg^-1）， with six mice in each group. After 12 weeks of continuous gavage， urine and kidney specimens were collected， and the 24-h urinary protein and the urinary albumin-to-creatinine ratio （UACR） in mice were measured. Hematoxylin-eosin （HE） staining， periodic acid-Schiff （PAS） staining， and Masson staining were performed for observation of histopathological changes in kidneys. Immunofluorescence assay was employed to detect the positive expression of the podocyte marker protein nephrin. Oil red O staining was used to detect renal lipid deposition. Enzyme linked immunosorbent assay was employed to measure the levels of interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） in the renal tissue. Western blot was employed to determine the expression levels of sterol regulatory element-binding protein cleavage-activating protein （SCAP）， sterol regulatory element-binding protein-1c （SREBP-1c）， and NOD-like receptor protein 3 （NLRP3） in the renal tissue. ResultsCompared with the blank group， the model group showed increases in 24-h urinary protein and UACR （P<0.05）， glomeruli exhibiting capsule adhesion， collagen fiber deposition， mesangial proliferation， and inflammatory cell infiltration， elevated levels of IL-1β， IL-6， and TNF-α （P<0.05）， reduced positive expression of nephrin （P<0.05）， increased lipid deposition （P<0.05）， and up-regulated expression of SCAP， SREBP-1c， and NLRP3 （P<0.05） in the renal tissue. Compared with the model group， the treatment with losartan potassium or high-dose Zhenzhu Tiaozhi capsules for 12 weeks decreased 24-h urinary protein and UACR （P<0.05）， and the treatment with low-dose Zhenzhu Tiaozhi capsules for 12 weeks reduced the 24-h urinary protein （P<0.05）. Pathological staining results revealed that kidney damage in mice from all treatment groups was alleviated， with reduced inflammatory infiltration， collagen fiber deposition， and mesangial proliferation， and increased positive expression of nephrin in the renal tissue （P<0.05）. In addition， all the treatment groups showed reduced lipid droplets （P<0.05）， lowered levels of IL-1β， IL-6， and TNF-α （P<0.05）， and down-regulated expression of SCAP， SREBP-1c， and NLRP3 （P<0.05） in the renal tissue. ConclusionZhenzhu Tiaozhi capsules can ameliorate kidney damage in the mouse model of diabetic kidney disease by inhibiting the activation of the SCAP-SREBP-1c/NLRP3 signaling pathway， which reduces renal lipid deposition and inflammation.

ObjectiveTo investigate the protective effects and mechanisms of Zhenzhu Tiaozhi capsules on the kidneys in the mouse model of diabetic kidney disease. MethodsThirty male C57BL/6J mice were selected as experimental objects. The model of diabetic kidney disease was induced by intraperitoneal injection of streptozotocin （STZ） at 40 mg·kg^-1 for 5 days combined with a high-fat diet （HFD）. Fasting blood glucose （FBG） ≥ 11.1 mmol·L^-1， increased urine volume， and continuous appearance of proteinuria indicated successful modeling. Mice were grouped as follows： Blank， model， low- and high-dose （0.98 and 1.96 g·kg^-1， respectively） Zhenzhu Tiaozhi capsules， and losartan potassium （30 mg·kg^-1）， with six mice in each group. After 12 weeks of continuous gavage， urine and kidney specimens were collected， and the 24-h urinary protein and the urinary albumin-to-creatinine ratio （UACR） in mice were measured. Hematoxylin-eosin （HE） staining， periodic acid-Schiff （PAS） staining， and Masson staining were performed for observation of histopathological changes in kidneys. Immunofluorescence assay was employed to detect the positive expression of the podocyte marker protein nephrin. Oil red O staining was used to detect renal lipid deposition. Enzyme linked immunosorbent assay was employed to measure the levels of interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） in the renal tissue. Western blot was employed to determine the expression levels of sterol regulatory element-binding protein cleavage-activating protein （SCAP）， sterol regulatory element-binding protein-1c （SREBP-1c）， and NOD-like receptor protein 3 （NLRP3） in the renal tissue. ResultsCompared with the blank group， the model group showed increases in 24-h urinary protein and UACR （P<0.05）， glomeruli exhibiting capsule adhesion， collagen fiber deposition， mesangial proliferation， and inflammatory cell infiltration， elevated levels of IL-1β， IL-6， and TNF-α （P<0.05）， reduced positive expression of nephrin （P<0.05）， increased lipid deposition （P<0.05）， and up-regulated expression of SCAP， SREBP-1c， and NLRP3 （P<0.05） in the renal tissue. Compared with the model group， the treatment with losartan potassium or high-dose Zhenzhu Tiaozhi capsules for 12 weeks decreased 24-h urinary protein and UACR （P<0.05）， and the treatment with low-dose Zhenzhu Tiaozhi capsules for 12 weeks reduced the 24-h urinary protein （P<0.05）. Pathological staining results revealed that kidney damage in mice from all treatment groups was alleviated， with reduced inflammatory infiltration， collagen fiber deposition， and mesangial proliferation， and increased positive expression of nephrin in the renal tissue （P<0.05）. In addition， all the treatment groups showed reduced lipid droplets （P<0.05）， lowered levels of IL-1β， IL-6， and TNF-α （P<0.05）， and down-regulated expression of SCAP， SREBP-1c， and NLRP3 （P<0.05） in the renal tissue. ConclusionZhenzhu Tiaozhi capsules can ameliorate kidney damage in the mouse model of diabetic kidney disease by inhibiting the activation of the SCAP-SREBP-1c/NLRP3 signaling pathway， which reduces renal lipid deposition and inflammation.

OBJECTIVE: To observe the effects of electroacupuncture (EA) on improving motor function and regulating microglial activation based on Notch receptor 1 (Notch1)/Hes family bHLH transcription factor 1 (Hes1) pathway in mice with Parkinson's disease (PD). METHODS: Thirty-six male C57BL/6 mice were randomly divided into a control group, a model group and an EA group, 12 mice in each group. PD model was established by intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) for 7 consecutive days in the model group and the EA group. From the 1st day of modeling, EA was applied at "Baihui" (GV20) and bilateral "Shenshu" (BL23) in the EA group, with continuous wave, in frequency of 2 Hz and current of 2 mA, 15 min a time, once a day for 14 days continuously. The behavioral performance was evaluated by gait test, pole climbing test and hanging test, the number of positive cells of tyrosine hydroxylase (TH) and the co-expression positive cells of Notch1/ionized calcium binding adaptor molecule 1 (Iba-1) in the substantia nigra of midbrain was assessed by immunofluorescence, the protein expression of TH, α-synuclein (α-syn), Notch1, Hes1, Iba-1, inducible nitric oxide synthase (iNOS), Arginase-1 (ARG1), tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 and IL-10 was detected by Western blot, the mRNA expression of Notch1 and Hes1 was detected by real-time PCR. RESULTS: Compared with the control group, in the model group, the stride frequency was accelerated (P<0.001) and the stride length was shortened (P<0.001) for the four limbs, the pole climbing test time was prolonged (P<0.01) and the grip level was reduced (P<0.01); in the substantia nigra of midbrain, the number of positive cells of TH was decreased (P<0.001), the number of co-expression positive cells of Notch1/Iba-1 was increased (P<0.001), the protein expression of α-syn, Notch1, Hes1, Iba-1, iNOS, TNF-α, IL-1βand IL-6 was increased (P<0.01, P<0.05, P<0.001), the protein expression of TH, ARG1 and IL-10 was decreased (P<0.01, P<0.001), the mRNA expression of Notch1 and Hes1 was increased (P<0.01). Compared with the model group, in the EA group, the stride frequency was decelerated (P<0.001) and the stride length was increased (P<0.05, P<0.01, P<0.001) for the four limbs, the pole climbing test time was shortened (P<0.05) and the grip level was increased (P<0.05); in the substantia nigra of midbrain, the number of positive cells of TH was increased (P<0.01), the number of co-expression positive cells of Notch1/Iba-1 was decreased (P<0.001), the protein expression of α-syn, Notch1, Hes1, Iba-1, iNOS, TNF-α, IL-6 and IL-1β was decreased (P<0.05, P<0.01), the protein expression of TH, ARG1 and IL-10 was increased (P<0.05, P<0.001, P<0.01), the mRNA expression of Notch1 and Hes1 was decreased (P<0.05). CONCLUSION EA can improve the behavioral performance and protect the dopaminergic neurons in PD mice, its mechanism may relate to the inhibition of Notch1/Hes1-mediated neuroinflammation, thus inhibiting the microglial activation.
Animals ; Electroacupuncture ; Microglia/metabolism* ; Male ; Receptor, Notch1/metabolism* ; Parkinson Disease/physiopathology* ; Transcription Factor HES-1/metabolism* ; Mice ; Mice, Inbred C57BL ; Humans ; Signal Transduction

Objective To screen and identify specific monoclonal antibodies targeting the surface envelope protein E8L of the monkeypox intracellular mature virus based on mRNA immunization and single-cell sequencing.Methods E8L-mRNA was synthesized in vitro and encapsulated into lipid nanoparticles(LNPs)for immunization to induce immune responses.The antibody level in blood serum was detected to find out when the mice had produced sufficient antibodies.B cells in the spleen of the mice were isolated by flow cytometry sorting,followed by library construction and sequencing.Based on the analysis of light and heavy chain sequences of the antibodies from the sequencing data,E8L candidate monoclonal antibodies were screened according to the abundance ranking and expressed.The affinity of these monoclonal antibodies was detected by enzyme-linked immunosorbent assay(ELISA).Candidate antibodies with high affinity for E8L were applied to immunochromatographic assays to detect the monkeypox virus(MPXV)E8L protein.Results Mice were immunized by lipid nanoparticles encapsulating E8L-mRNA.Approximately 1.5 million B cells were selected by flow cytometry.After single-cell sequencing,17 candidate monoclonal antibodies for E8L were identified and expressed.Two high affinity monoclonal antibodies were obtained for monkeypox virus E8L protein by ELISA.These two antibodies were used as the basis for immunochromatographic assays to detect E8L protein,with a sensitivity of 0.5 ng/mL.Conclusion Two monoclonal antibodies with high affinity for the MPXV E8L protein are obtained,which can be potentially used for detecting monkeypox virus.

Background::An evidence gap still exists regarding the efficacy and safety of tegoprazan in patients with erosive esophagitis (EE) in China. This study aimed to verify the efficacy and safety of tegoprazan vs. esomeprazole in patients with EE in China. Methods::This study was a multicenter, randomized, double-blind, parallel, active-controlled, non-inferiority phase III trial of patients with EE randomized 1:1 to tegoprazan 50 mg/day vs. esomeprazole 40 mg/day. This study was conducted in 32 sites between October 24, 2018 and October 18, 2019. The primary endpoint was the cumulative endoscopic healing rate at week 8. The secondary endpoint included endoscopic healing rate at week 4, changes in the reflux disease questionnaire (RDQ) and gastroesophageal reflux disease health-related quality of life (GERD-HRQL) scores, and symptom improvement. Results::A total of 261 patients were randomized: 132 to the tegoprazan group and 129 to the esomeprazole group. The cumulative endoscopic healing rate at 8 weeks in the tegoprazan group was non-inferior to that of the esomeprazole group (91.1% vs. 92.8%, difference: -1.7%, 95% confidence interval [CI]: -8.5%, 5.0%, P = 0.008). There were no statistically significant differences in the changes in RDQ (total, severity, and frequency) and GERD-HRQL scores between the two groups (all P >0.05). The percentages of days without symptoms, including daytime and nighttime symptoms based on patients' diaries, were similar between the two groups (all P >0.05). In the tegoprazan and esomeprazole groups, 71.5% (93/130) and 61.7% (79/128) of the participants reported adverse events (AEs), 2.3% and 0 experienced serious AEs, while 70.0% and 60.2% had treatment-emergent AEs, respectively. Conclusion::Tegoprazan 50 mg/day demonstrated non-inferior efficacy in healing EE, symptom improvement, and quality of life, and it has similar tolerability compared with esomeprazole 40 mg/day.

Objective:To analyze the association between inflammation-related dietary patterns and the risk for cognitive impairment in older adults aged ≥65 years in longevity areas in China by using reduced rank regression (RRR) analysis.Methods:This study used cross-sectional data from the 2021 Healthy Aging and Biomarkers Cohort Study, including the information about study participants' demographic characteristics, lifestyles, daily life activities, and disease histories. Dietary intake was obtained by using a simplified food frequency questionnaire. Cognitive impairment was evaluated based on the Mini-Mental State Examination Scale combined with years of education. Fasting venous blood samples were collected to detect inflammatory markers, especially high-sensitivity C-reactive protein (hs-CRP) and the platelet-to-lymphocyte ratio (PLR). RRR analysis was used to obtain inflammation-related dietary patterns using hs-CRP and PLR as response variables. Multivariate logistic regression model was used to analyze the association between dietary pattern score and the risk for cognitive impairment. Restricted cubic spline was used to explore the dose response relationship, and mediation analysis was used to quantify the mediating effects of hs-CRP and PLR.Results:Two dietary patterns were identified with RRR. The primary pattern was characterized by higher intakes of flour, red meat, and dairy products, and lower intake of fresh vegetables, explaining 6.84% of the variance in food intake and 0.50% of the variance in inflammatory markers. Compared with the T1 group, the T3 group had significantly higher risk for cognitive impairment ( OR=1.242, 95% CI: 1.034-1.491). Each one standard deviation increase in the dietary pattern score was associated with an 8.7% increase in the risk for cognitive impairment ( OR=1.087, 95% CI: 1.008-1.172), with a significant linear trend (overall-model P<0.001, non-linear P=0.295). Mediation analysis indicated that hs-CRP mediated 6.2% of the association between the dietary pattern and the risk for cognitive impairment. Conclusion:The inflammation- related dietary pattern characterized by higher consumption of flour, red meat, and dairy products and lower consumption of fresh vegetables is associated with an increased risk for cognitive impairment in older adults, and hs-CRP partially mediates this association.

Objective:To observe the safety and short-term efficacy of using an autologous fascia lata sling (AFLS) for tension-free mid-urethral sling (MUS) in the treatment of stress urinary incontinence (SUI).Methods:Between February 2022 and December 2023, 11 patients with SUI underwent AFLS-MUS. Preoperative data were recorded, including basic patient information and completion of urinary distress inventory 6 (UDI-6). During surgery, AFLS was harvested through a small incision using a tendon extractor, and used as a sling for transobturator or retropubic MUS. Perioperative indicators were recorded, including surgical approach, operation time, intraoperative blood loss, postoperative hospital stay, duration of catheterization, perioperative complications (Clavien-Dindo classification), and surgical costs. Follow-ups included outpatient physical examination at 2 months postoperatively, and telephone follow-up at 6 months, 1 year, and annually thereafter. Follow-up content included the presence or absence of urinary leakage symptoms, UDI-6, satisfaction, patient global impression of improvement (PGI-I), and complications.Results:The age of the 11 patients was (54.8±10.9) years (range: 41-72 years), with body mass index of (23.9±1.8) kg/m2 (range: 21.4-27.3 kg/m2). All patients experienced urinary leakage after coughing, sneezing and physical activity, with positive SUI provocation tests. The preoperative UDI-6 was 50.0±21.6 (range: 16.7-79.2), the result of 1-hour pad test was (18.9±12.0) g (range: 2.5-71.2 g). Four cases underwent MUS only, with operation time of (98.0±13.3) minutes (range: 86-117 minutes), and intraoperative blood loss of (17.5±5.0) ml (range: 10-20 ml); 7 cases also underwent pelvic floor repair simultaneously. The postoperative hospital stay was (3.5±2.0) days (range: 2-9 days). The duration of catheterization was (4.5±3.8) days (range: 2-11 days), with postoperative urinary retention in three cases, one of which underwent sling release surgery due to severe postoperative voiding difficulty 1 week after MUS, with no other complications of Clavien-Dindo grade 2 or above. The cost of AFLS harvest plus MUS was (2 762±293) yuan. At the 2-month outpatient follow-up, all patients were free of urinary leakage symptoms, with UDI-6 of 2.3±1.9 (range: 0-8.3); satisfaction was "very satisfied" in 10 cases and "fairly satisfied" in 1 case, with PGI-I all being "much better", and pelvic examinations were normal. Telephone follow-up showed one case lost to follow-up, and the remaining 10 cases had follow-up time of (18.6±4.9) months (range: 7-26 months), all without urinary leakage, with UDI-6 of 2.7±2.6, satisfaction rated as "very satisfied", and PGI-I all "much better".Conclusion:This modified AFLS-MUS for the treatment of SUI shows good short-term efficacy and high safety in harvest site, with the need for more data accumulation and long-term follow-up.

Three-dimensional(3D)printing technology,which is based on computer aided design,presents innovative solutions for clinical surgical treatments and is increasingly being utilized in clinical settings.By integrating medical-image processing software and a 3D printer with customized materials,physicians can efficiently fabricate a 3D model layer by layer,thus realizing the reconstruction of human-body models with specific objectives.These models vividly showcase the anatomical structures,fracture types,and rupture severity,thereby facilitating doctors in understanding the fracture being examined.Beyond serving as a mere demonstration tool,the physi-cal model assists doctors in designing personalized implants and selecting customized surgical guide plates and fixation screws.Surgeons can improve the surgical accuracy and efficacy by refining the surgical plan via preoperative simulation operations.This article presents a summary of the application of computer-simulation surgery in conjunction with 3D printing technology in various complex surgeries.Addi-tionally,the significant potential applications of 3D printing technology in the clinical orthopedics field are discussed.

Objective:To analyze the association between inflammation-related dietary patterns and the risk for cognitive impairment in older adults aged ≥65 years in longevity areas in China by using reduced rank regression (RRR) analysis.Methods:This study used cross-sectional data from the 2021 Healthy Aging and Biomarkers Cohort Study, including the information about study participants' demographic characteristics, lifestyles, daily life activities, and disease histories. Dietary intake was obtained by using a simplified food frequency questionnaire. Cognitive impairment was evaluated based on the Mini-Mental State Examination Scale combined with years of education. Fasting venous blood samples were collected to detect inflammatory markers, especially high-sensitivity C-reactive protein (hs-CRP) and the platelet-to-lymphocyte ratio (PLR). RRR analysis was used to obtain inflammation-related dietary patterns using hs-CRP and PLR as response variables. Multivariate logistic regression model was used to analyze the association between dietary pattern score and the risk for cognitive impairment. Restricted cubic spline was used to explore the dose response relationship, and mediation analysis was used to quantify the mediating effects of hs-CRP and PLR.Results:Two dietary patterns were identified with RRR. The primary pattern was characterized by higher intakes of flour, red meat, and dairy products, and lower intake of fresh vegetables, explaining 6.84% of the variance in food intake and 0.50% of the variance in inflammatory markers. Compared with the T1 group, the T3 group had significantly higher risk for cognitive impairment ( OR=1.242, 95% CI: 1.034-1.491). Each one standard deviation increase in the dietary pattern score was associated with an 8.7% increase in the risk for cognitive impairment ( OR=1.087, 95% CI: 1.008-1.172), with a significant linear trend (overall-model P<0.001, non-linear P=0.295). Mediation analysis indicated that hs-CRP mediated 6.2% of the association between the dietary pattern and the risk for cognitive impairment. Conclusion:The inflammation- related dietary pattern characterized by higher consumption of flour, red meat, and dairy products and lower consumption of fresh vegetables is associated with an increased risk for cognitive impairment in older adults, and hs-CRP partially mediates this association.

Objective:To observe the safety and short-term efficacy of using an autologous fascia lata sling (AFLS) for tension-free mid-urethral sling (MUS) in the treatment of stress urinary incontinence (SUI).Methods:Between February 2022 and December 2023, 11 patients with SUI underwent AFLS-MUS. Preoperative data were recorded, including basic patient information and completion of urinary distress inventory 6 (UDI-6). During surgery, AFLS was harvested through a small incision using a tendon extractor, and used as a sling for transobturator or retropubic MUS. Perioperative indicators were recorded, including surgical approach, operation time, intraoperative blood loss, postoperative hospital stay, duration of catheterization, perioperative complications (Clavien-Dindo classification), and surgical costs. Follow-ups included outpatient physical examination at 2 months postoperatively, and telephone follow-up at 6 months, 1 year, and annually thereafter. Follow-up content included the presence or absence of urinary leakage symptoms, UDI-6, satisfaction, patient global impression of improvement (PGI-I), and complications.Results:The age of the 11 patients was (54.8±10.9) years (range: 41-72 years), with body mass index of (23.9±1.8) kg/m2 (range: 21.4-27.3 kg/m2). All patients experienced urinary leakage after coughing, sneezing and physical activity, with positive SUI provocation tests. The preoperative UDI-6 was 50.0±21.6 (range: 16.7-79.2), the result of 1-hour pad test was (18.9±12.0) g (range: 2.5-71.2 g). Four cases underwent MUS only, with operation time of (98.0±13.3) minutes (range: 86-117 minutes), and intraoperative blood loss of (17.5±5.0) ml (range: 10-20 ml); 7 cases also underwent pelvic floor repair simultaneously. The postoperative hospital stay was (3.5±2.0) days (range: 2-9 days). The duration of catheterization was (4.5±3.8) days (range: 2-11 days), with postoperative urinary retention in three cases, one of which underwent sling release surgery due to severe postoperative voiding difficulty 1 week after MUS, with no other complications of Clavien-Dindo grade 2 or above. The cost of AFLS harvest plus MUS was (2 762±293) yuan. At the 2-month outpatient follow-up, all patients were free of urinary leakage symptoms, with UDI-6 of 2.3±1.9 (range: 0-8.3); satisfaction was "very satisfied" in 10 cases and "fairly satisfied" in 1 case, with PGI-I all being "much better", and pelvic examinations were normal. Telephone follow-up showed one case lost to follow-up, and the remaining 10 cases had follow-up time of (18.6±4.9) months (range: 7-26 months), all without urinary leakage, with UDI-6 of 2.7±2.6, satisfaction rated as "very satisfied", and PGI-I all "much better".Conclusion:This modified AFLS-MUS for the treatment of SUI shows good short-term efficacy and high safety in harvest site, with the need for more data accumulation and long-term follow-up.